• Title/Summary/Keyword: Lysosomal enzyme activity

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Subcellular Localization of Capsaicin-Hydrolyzing Enzyme in Rat Hepatocytes (Capsaicin 가수분해효소의 흰쥐 간세포내 소재확인)

  • Park, Young-Ho;Lee, Sang-Sup
    • YAKHAK HOEJI
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    • v.38 no.1
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    • pp.12-19
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    • 1994
  • Capsaicin(8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component of Capsicum fruits. This work is directed to the capsaicin-hydrolyzing enzyme playing a key role in the rate limiting and critical step of capsaicin metabolism. In order to get precise information on the enzyme's subcellular location, rat liver homogenate was divided into six subcellular fractions by differential centrifugation technique: crude nuclear pellet, PNS(post nuclear supernatant) fraction, lysosomal pellet, cytosol, Tris wash fraction, micrisomes. Capsaicin-hydrolysing enzyme activity was analysed by high performance liquid chromatography(HPLC). This enzyme was found at the highest specific activity in the microsomal fraction and co-distributed with marker enzymes of the endoplasmic reticulum, NADPH-cytochrome c reductase and nucleoside diphosphatase. This is compatible with the result of ninhydrin color reaction of vanillylamine, primary metabolite of capsaicin hydrolysis, on thin layer chromatography(TLC). This enzyme is most active at pH $8.0{\sim}9.0$. Definite subcellular location of this enzyme will make it easy to proceed with further study.

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Immune Cell Stimulating Activity of Wheat Arabinoxylan (밀 arabinoxylan의 면역세포 활성화 작용)

  • Choi, Eun-Mi;Lim, Tae-Soo;Lee, Hye-Lim;Hwang, Jae-Kwan
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.510-517
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    • 2002
  • Effects of wheat arabinoxylan on mouse spleen lymphocytes and peritoneal macrophages were examined in vitro. Among three wheat arabinoxylans (A1: low MW, A2: medium MW, A3: high MW), A3$(50{\sim}100\;{\mu}g/mL)$ increased the viability of spleen lymphocytes up to $114{\sim}125%$ of the control. A1 and A3 $(20\;{\mu}g/mL)$ increased the viability of lipopolysaccharide-treated lymphocytes synergistically. Viability of murine peritoneal macrophages treated with wheat arabinoxylans $(10{\sim}100{\mu}g/mL)$ was increased up to $135{\sim}175%$ of the control. The cytotoxic activity of macrophages against murine lymphocytic leukemic cell increased in the presence of wheat arabinoxylan. Phagocytic index of macrophages treated with wheat arabinozylans $(20\;{\mu}g/mL)$ significantly increased $197{\sim}232%$ compared with the control, and lysosomal phosphatase and myeloperoxidase activities also increased significantly (p<0.05). Treatment of wheat arabinoxylans tended to decrease nitrite production, but significantly stimulated $H_2O_2\;and\;O_2$ productions of macrophages (p<0.05). These results indicate that the immunostimulating effect of wheat arabinoxylan may be closely related with lysosomal enzyme activity and reactive oxygen intermediate production of macrophages.

Effects of Soyosangamibang on antitumor chemotherapy and lysosomal enzymes of tumor cel (소요산가미방(逍遙散加味方)이 항암화학요법제(抗癌化學療法劑)의 항종양효과(抗腫瘍效果) 및 종양세포(腫瘍細胞)의 lysosomal enzymes에 미치는 영향(影響))

  • Cho, Hyun-Ju;Won, Bong-Hui;Mun, Gu;Mun, Seok-Jae;Jeon, Byeong-Hun
    • The Journal of Korean Medicine
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    • v.18 no.2
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    • pp.119-136
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    • 1997
  • In order to investigate the effects of Soyosangamibang Extract(逍遙散加味方抽出液) on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 cells after exposure to the extract of Soyosangamibang extract depressed the growth of tumor cells by concentration of Soyosangamibang, 2. Antitumor activity of the ethanol extract from Soyosangamibang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of 200mg/kg and MMC 0.1mg/kg is improved Over 50%. 3. WhenSoyosangamibang extract and MMC are administrated together, the weight of turnor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Soyosangarmibang extract and MMC are more significantly improved than MMC alone. According to the above results, it could be suggested that Soyosangamibang extract has indirect antitumor effect by strengthen the effect of MMC.

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Prevention of Ischemic Damage in Working Rat Hearts by Calcium Channel Blocker and Calmodulin Inhibitors (흰쥐심장의 허혈손상에 대한 Calcium 통로봉쇄제와 Calmodulin 억제제의 예방효과에 대한 연구)

  • 성시찬
    • Journal of Chest Surgery
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    • v.22 no.6
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    • pp.901-913
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    • 1989
  • This study was investigated under the postulation that activation of intracellular calcium- calmodulin complex during ischemia-reperfusion leads to myocardial injury. The protective effects of calcium channel blocker, diltiazem and calmodulin inhibitors, trifluoperazine, flunarizine and calmidazolium from ischemic injury in rat hearts were observed by using Langendorff apparatus when the antagonists were infused for 3 min in the beginning of ischemia. Thereby, an increase in resting tension developed during 30-min ischemia was analyzed with regard to [1] the degree of cardiac functional recovery following 60-min reperfusion, [2] changes in biochemical variables evoked during 30-min ischemia. The results obtained were as follows: l. In the ischemic group, the resting tension was increased by 4.1*0.2 g at 30-min ischemia. However, the increase in resting tension was markedly reduced not only by pretreatment with diltiazem [3.3 p M] but also with calmodulin inhibitors, trifluoperazine [3.3 p M], flunarizine [0.5 p M] and calmidazolium [0.5 p M], respectively. 2. Recovery of myocardial contractility, +dF /dt and coronary flow were much reduced when evoked by reperfusion in the ischemic group. These variables were significantly improved either by pretreatment with diltiazem or with calmodulin inhibitors. 3. The resting tension increment evoked during ischemia was significantly inversely correlated with the degree of cardiac function recovered during reperfusion. 4. Following 30-min ischemia, the production of malondialdehyde and release of lysosomal enzyme were much increased in association with a decrease in creatine kinase activity. 5. The increases in malondialdehyde production and release of free lysosomal enzyme were suppressed by pretreatment with calmodulin inhibitors as well as diltiazem. Likewise, the decrease of creatine kinase activities was prevented by these calcium antagonists. With these results, it is indicated that a increase in resting tension observed during ischemia has an inverse relationship to the cardiac function recovered following reperfusion, and further, the later may be significantly dependent on the degree of biochemical alterations occurred during ischemia such as decrease in creatine kinase activity, increased production of malondialdehyde and increased release of free lysosomal enzyme. Thus it is concluded that calmodulin plays a pivotal role in the process of ischemic injury.

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High Calcium in the Diet and Vitamin D Inhibit a Bone Matrix Degrading Enzyme $\beta$-Glucuronidase Activity

  • Ha, Kyung-Sun;Yook, Hong-Sun;Kang, Il-Jun;Han, Eun-Kyung;Kim, Hyun-Sook;Chung, Cha-Kwon
    • Preventive Nutrition and Food Science
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    • v.2 no.1
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    • pp.55-60
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    • 1997
  • A lysosomal and matrix degrading enzyme $\beta$-glucuronidase activity was measured in BALS/c mice fed high and low Ca in combination with the i.p. adminstration of calcium-regulating hormones including parathyoid hormone(PTH), calcitonin(CT) and cholecalciferol(Vit D). After feeding experimental diets for five weeks, mice were sacrificed by cervical dislocation and the enzyme was fluometrically measured at 440nm. $\beta$-Glucuronidase activity was inhibited by high calcium in the diet. in addition, vitamin D also inhibited the enzyme activity in the serum regardless of the level of dietary calcium. In contrast, PTH has shown to stimulate the enzyme at all the levels of dietary calcium. Calcitonin, and inhibitor of PTH action for bone resorption, revealed to curb PTH effect in this enzyme, whereas CT stimulated the action of vitamin D in the serum. The above results led us to conclude that osteoclastic bone resorption and senile osteoporosis may be reduced by adequate dietary calcium and vitamin D.

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Antitumor Effects and Immunomodulating Activities of Phellinus linteus Extract in a CT-26 Cell-Injected Colon Cancer Mouse Model

  • Yang, Byung-Keun;Hwang, Seung-Lark;Yun, Ik-Jin;Do, Eun-Ju;Lee, Won-Ha;Jung, Young-Mi;Hong, Sung-Chang;Park, Dong-Chan
    • Mycobiology
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    • v.37 no.2
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    • pp.128-132
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    • 2009
  • The antitumor effects of Phellinus linteus extract (Keumsa Linteusan) were investigated in a CT-26 cell-injected colon cancer mouse model. When administered orally (250${\sim}$1,000 mg/kg body weight), Keumsa Linteusan significantly inhibited the growth of solid colon cancer. The highest dose was highly effective, reducing tumor formation by 26% compared with the control group. The anticomplementary activity of Keumsa Linteusan increased in a dose-dependent manner. Lysosomal enzyme activity of macrophages was increased by 2-fold (100 ${\mu}$/ml) compared with the control group. Keumsa Linteusan can be regarded as a potent enhancer of the innate immune response, and can be considered as a very promising candidate for antitumor action.

Enzyme activity changes by intraperitoneal injection of uranium in the carp liver (우라늄 투여후 간조직에서의 효소활성도의 변화)

  • Kim, In-Gyu;Kim, Kug-Chan;Kim, Jin-Kyu;Kim, Sang-Bok;Chun, Ki-Chung;Park, Hyo-Kook;Lee, Kang-Suk
    • Journal of Radiation Protection and Research
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    • v.18 no.2
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    • pp.61-69
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    • 1993
  • We examined various enzyme activity changes by intraperitoneal injection uranium in the carp liver. These enzyme activity changes can be used as biochemical indicators of internal exposure to uranium. The results were followings ; 1) Total protein concentration decreased by intraperitoneal injection in the carp liver. 2) Lysosomal acid pretense and ${\beta}-glucuronidase$ activities increased in the liver until sixth intraperitoneal injection of uranium, but Lysosomal acid phosphatase activities decreased in the liver until the sixth injection of uranium. 3) Alkaline phosphatase activities sharply increased and Glutamate oxaloacetate Transaminase activities steadily decreased in the liver until the sixth injection of uranium. 4) Creatine %kinase activities steadily decreased and malate dehydrogenase activities sharply decreased in the liver after the primary injection of uranium. Any malate dehydrogenase activities was not detected after sixth injection of uranium.

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A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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Joint Problems in Patients with Mucopolysaccharidosis Type II

  • Kim, Min-Sun;Kim, Jiyeon;Noh, Eu Seon;Kim, Chiwoo;Cho, Sung Yoon;Jin, Dong-Kyu
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.5 no.1
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    • pp.17-21
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    • 2021
  • Hunter syndrome or mucopolysaccharidosis type II (MPS-II) (OMIM 309900) is a rare lysosomal storage disorder caused by deficiency in the activity of the enzyme iduronate-2-sulfatase. This enzyme is responsible for the catabolism of the following two different glycosaminoglycans (GAGs): dermatan sulfate and heparan sulfate. The lysosomal accumulation of these GAG molecules results in cell, tissue, and organ dysfunction. Patients can be broadly classified as having one of the following two forms of MPS II: a severe form and an attenuated form. In the severe form of the disease, signs and symptoms (including neurological impairment) develop in early childhood, whereas in the attenuated form, signs and symptoms develop in adolescence or early adulthood, and patients do not experience significant cognitive impairment. The involvement of the skeletal-muscle system is because of essential accumulated GAGs in joints and connective tissue. MPS II has many clinical features and includes two recognized clinical entities (mild and severe) that represent two ends of a wide spectrum of clinical severities. However, enzyme replacement therapy is likely to have only a limited impact on bone and joint disease based on the results of MPS II studies. The aim of this study was to review the involvement of joints in MPS II.

Longterm Benefit and Unmet Needs in Enzyme Replacement Therapy of Gaucher (고셔병 환자에서 효소대체치료의 장기적 효과와 한계)

  • Kim, Yoo-Mi
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.1
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    • pp.42-47
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    • 2014
  • Gaucher disease is a multisystemic disorder arising from a deficient activity of the lysosomal enzyme glucocerebrosidase, which leads to accumulation of glycosylceraide and other glycolipids in the regiculoendothelial system. The characteristics of Gaucher disease are anemia, thrombocytopenia, hepatosplenomegaly, and skeletal disease. Enzyme replacement therapy (ERT) has been proven to prevent progressive manifestations of Gaucher disease and effective in improving anemia, thrombocytopenia, bone markers and biomarkers. However, some patient needs still remain unmet because of the inaccessibility of certain sites including brain, bone and various organs. ERT could not Improve the irreversible lesion such as liver fibrosis, hepatopulmonary syndrome, and necrosis or infarction of bone and other organs. Adult patients with Gaucher disease should be screened for longterm complication such as bone disease, pulmonary hypertension, gallstone, and cancer, especially in patients with splenectomy. Parkinsonism and polyneuropathy was also reported among patients with type 1 Gaucher disease, but ERT does not improve neurological function. We need to review the benefits and unmet needs of ERT in Gaucher disease.