• Title/Summary/Keyword: survival enzyme

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A Study on Anti-aning of Electroacupuncture at Joksamni($ST_{36}$) in Rats (족삼리(足三里) 전침(電鍼)이 흰쥐의 노화(老化)에 미치는 영향(影響))

  • Lee, Jun-Seong;Yim, Yun-Kyoung;Kim, Young-Il
    • Journal of Acupuncture Research
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    • v.24 no.3
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    • pp.145-161
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    • 2007
  • Objectives : The purpose of this study is to investigate the effect of electroacupuncture at Joksanmi($ST_{36}$) on Rat's aging by experimental methods. Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. First, I measured the cell survival rate of the mice lung fibroblasts. Second, the incidence rate of arthritis and arthritis index of CIA were analyzed. Third, the levels of IL-6, $INF-{\gamma}$ , $INF-{\alpha}$ , $IL-1{\beta}$, IgG, IgM and anti-collagen II were measured in serum and the level of $INF-{\gamma}$, $INF-{\gamma}$/IL-4 ratio in the CIA mouse spleen cell culture. Fourth, histological analysis of the mice joint was performed. Fifth, the expression ratio of $CD3e^+$ cells to $CD19^+$ cells, $CD4^+$ cells to $CD8^+$ cells, $CD11a^+CD19^+$ cells, $CD4^+CD25^+$ cells, $CD3^+CD69^+$ cells, and $CD11b^+Gr-1^+$ cells were checked. Results: 1. The hematocrit was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 2. The glucose, ALP were meaningfully (p<0.05, p<0.05) decreased in $ST_{36}$ group in comparison with the control group. 3. In the activation of hepatic antioxidase, the catalase was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the control group. 4. In the liver, the NO formation was meaningfully (p<0.001) decreased in $ST_{36}$ group in comparison with the nomal, control and sham group. 5. In the activation of splenetic antioxidase, the GSH was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the sham group, and the catalase was meaningfully (p<0.001) increased in $ST_{36}$ group in comparison with the control group. 6. In the spleen, the MDA formation was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 7. In the spleen cell culture, the levels of IL-6 was meaningfully (p<0.05) decreased in comparison with the sham group, $INF-{\gamma}$( was meaningfully (p<0.05) increased in comparison with the control group. Conclusions: According to the results, it is considered that electroacupuncture at Joksamni($ST_{36}$) is effective in increasing the activities of antioxidative enzyme and inhibiting lipid peroxides, and has an immunomodulatory reaction. In conclusion, it han an significant anti-aging effect and need more experimental study.

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Protective Effect of Nitric Oxide against Oxidative Stress under UV-B Radiation in Maize Leaves (UV-B 조사시 옥수수 잎의 산화적 스트레스에 대한 Nitric Oxide의 보호효과)

  • Kim, Tae-Yun;Jo, Myung-Hwan;Hong, Jung-Hee
    • Journal of Environmental Science International
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    • v.19 no.12
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    • pp.1323-1334
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    • 2010
  • The effect of nitric oxide (NO) on antioxidant system and protective mechanism against oxidative stress under UV-B radiation was investigated in leaves of maize (Zea mays L.) seedlings during 3 days growth period. UV-B irradiation caused a decrease of leaf biomass including leaf length, width and weight during growth. Application of NO donor, sodium nitroprusside (SNP), significantly alleviated UV-B stress induced growth suppression. NO donor permitted the survival of more green leaf tissue preventing chlorophyll content reduction and of higher quantum yield for photosystem II than in non-treated controls under UV-B stress, suggesting that NO has protective effect on chloroplast membrane in maize leaves. Flavonoids and anthocyanin, UV-B absorbing compounds, were significantly accumulated in the maize leaves upon UV-B exposure. Moreover, the increase of these compounds was intensified in the NO treated seedlings. UV-B treatment resulted in lipid peroxidation and induced accumulation of hydrogen peroxide ($H_2O_2$) in maize leaves, while NO donor prevented UV-B induced increase in the contents of malondialdehyde (MDA) and $H_2O_2$. These results demonstrate that NO serves as antioxidant agent able to scavenge $H_2O_2$ to protect plant cells from oxidative damage. The activities of two antioxidant enzymes that scavenge reactive oxygen species, catalase (CAT) and ascorbate peroxidase (APX) in maize leaves in the presence of NO donor under UV-B stress were higher than those under UV-B stress alone. Application of 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3- oxide (PTIO), a specific NO scavenger, to the maize leaves arrested NO donor mediated protective effect on leaf growth, photosynthetic pigment and free radical scavenging activity. However, PTIO had little effect on maize leaves under UV-B stress compared with that of UV-B stress alone. $N^{\omega}$-nitro-L-arginine (LNNA), an inhibitor of nitric oxide synthase (NOS), significantly increased $H_2O_2$ and MDA accumulation and decreased antioxidant enzyme activities in maize leaves under UV-B stress. This demonstrates that NOS inhibitor LNNA has opposite effects on oxidative resistance. From these results it is suggested that NO might act as a signal in activating active oxygen scavenging system that protects plants from oxidative stress induced by UV-B radiation and thus confer UV-B tolerance.

Enhanced Radiosensitivity of Tumor Cells Treated with Vanadate in Vitro (Vanadate 처리가 종양세포의 방사선 감수성에 미치는 영향)

  • Lee, Myung-Za;Lee, Won-Young
    • Radiation Oncology Journal
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    • v.12 no.2
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    • pp.129-141
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    • 1994
  • Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cellls but not CCL-120 normal cells to radiation. Ouabain inhibits the $Na^+-K^+$-pump rapidly thus it increases intracellular Na concentration, Vanadate which is distributed extensively in almost all living organisms is known to be a $Na^+-K^+$-ATPase inhibitors, This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of $Na^+-K^+$ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMC cells and frypan blue dye exclusion test for L120, and spleen cells. Measurements of $Na^+-K^+$-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined $10^{-6}M$ vanadate and radiation treated cells were done. The results were summerized as fellows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Mininum or no cytotoxicity was seen with vanadate below concentration of $10^{-6}M$. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. e. 2- Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. $Na^+-K^+$-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiaiton itself inhibited $Na^+-K^+$-ATPase activity of tumor cell with high $Na^+-K^+$-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with orginal $Na^+-K^+$-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on $Na^+-K^+$-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.

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Differential herbicide response of sulfonylurea-resistant Monochoria vagnalis accessions to sulfonylurea herbicides (서로 다른 지역에서 채집된 Sulfonylurea계 제초제 저항성 물달개비의 제초제 반응 차이)

  • Park, Tae-Seon;Lee, In-Yong;Park, Jae-Eup;Oh, Se-Mun
    • The Korean Journal of Pesticide Science
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    • v.11 no.4
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    • pp.269-275
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    • 2007
  • Four sulfonylurea(SU)-resistant Monochoria vaginalis(M. vaginalis) accessions were tested for levels of resistance to four SU herbicides which have been widely using in paddy fields of Korea, based on whole plant response and sensitivity of the target enzyme, acetolactate synthase(ALS). The resistant Naju, Nonsan and Gimje accessions were not affected to the survival by treatment with recommended dose of all SU herbicides tested. The $GR_{50}$ values for the Naju, Nonsan and Gimje accessions were 8- to 33-fold, 8- to 30-fold and 7- to 32-fold higher to recommended doses of all SU herbicides tested than the susceptible Cheongdo accession, respectively. However, the $GR_{50}$ values for Kimhae accession displayed an intermediate response and was only 4-to 13-fold more resistant than the susceptible accession. The ALS $I_{50}$ values for the Naju, Nonsan and Gimje accessions were 25- to 66-fold, 9- to 26-fold and 10- to 24-fold higher to recommended doses of all SU herbicides tested than the susceptible Cheongdo accession, respectively. However, the $I_{50}$ value for Kimhae accession was 4- to 9-fold more resistant than the susceptible accession, as determined by $I_{50}$ values of ALS.

Osteopontin Levels in Patients with Papillary Thyroid Cancer According to the Presence of Hashimoto's Thyroiditis

  • Park, So-Hyun;Park, Chan-Sung;Kim, Young-Il;Nam-Goong, Il-Seong;Kim, Yon-Seon;Lee, Jong-Cheol;Choi, Jung-Il;Park, Jeong-Woo;Kim, Eun-Sook
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2447-2451
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    • 2015
  • Background: Human papillary thyroid carcinoma (PTC) is often associated with Hashimoto's thyroiditis (HT); their coexistence improves PTC prognosis. Osteopontin, a secreted glycoprotein, plays a role in cell survival, immunity, and tumor progression, its expression being associated with a poor prognosis and metastasis in several malignancies. Osteopontin overexpression correlates with aggressive clinicopathological features in PTC. Lymph node metastases and large tumor size positively correlate with osteopontin positivity. This study aimed to: (1) confirm osteopontin overexpression in human PTC samples; (2) compare osteopontin expression levels in PTC cases with and without HT; and (3) identify correlations between tumor aggressiveness and osteopontin expression levels. Materials and Methods: Plasma osteopontin was assessed in 45 patients with PTC, 22 patients with PTC and HT, and 24 healthy controls by enzyme-linked immunosorbent assay. Thyroid tissue osteopontin mRNA and protein levels were analyzed by reverse transcription-polymerase chain reaction and Western blotting, respectively. Results: Plasma osteopontin levels were significantly higher in PTC patients than in healthy controls. Plasma osteopontin, tissue osteopontin mRNA, and tissue osteopontin protein levels were significantly lower in patients with PTC and HT than in those with PTC alone. In advanced disease stage cases, osteopontin mRNA and protein expression levels were lower in patients with PTC and HT than in those with PTC alone. However, the osteopontin expression level was not significantly associated with the TNM stage. Conclusions: Plasma osteopontin, tissue osteopontin mRNA, and tissue osteopontin protein levels were significantly lower in patients with PTC and HT than in those with PTC alone, suggesting that HT attenuates PTC aggressiveness through negative regulation of osteopontin expression.

Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

Cooperative Interactions between Toll-Like Receptor 2 and Toll-Like Receptor 4 in Murine Klebsiella pneumoniae Infections

  • Jeon, Hee-Yeon;Park, Jong-Hyung;Park, Jin-Il;Kim, Jun-Young;Seo, Sun-Min;Ham, Seung-Hoon;Jeong, Eui-Suk;Choi, Yang-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1529-1538
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    • 2017
  • Klebsiella pneumoniae is an opportunistic and clinically significant emerging pathogen. We investigated the relative roles of Toll-like receptor (TLR) 2 and TLR4 in initiating host defenses against K. pneumoniae. TLR2 knockout (KO), TLR4 KO, TLR2/4 double KO (DKO), and wild-type (WT) mice were inoculated with K. pneumoniae. Mice in each group were sacrificed after either 12 or 24h, and the lungs, liver, and blood were harvested to enumerate bacterial colony-forming units (CFU). Cytokine and chemokine levels were analyzed using enzyme-linked immunosorbent assay and real-time PCR, and pneumonia severity was determined by histopathological analysis. Survival was significantly shortened in TLR4 KO and TLR2/4 DKO mice compared with that of WT mice after infection with $5{\times}10^3CFU$. TLR2 KO mice were more susceptible to infection than WT mice after exposure to a higher infectious dose. Bacterial burdens in the lungs and liver were significantly higher in TLR2/4 DKO mice than in WT mice. Serum $TNF-{\alpha}$, MCP-1, MIP-2, and nitric oxide levels were significantly decreased in TLR2/4 DKO mice relative to those in WT mice, and TLR2/4 DKO mice showed significantly decreased levels of $TNF-{\alpha}$, IL-6, MCP-1, and inducible nitric oxide synthase mRNA in the lung compared with those in WT mice. Collectively, these data indicate that TLR2/4 DKO mice were more susceptible to K. pneumoniae infection than single TLR2 KO and TLR4 KO mice. These results suggest that TLR2 and TLR4 play cooperative roles in lung innate immune responses and bacterial dissemination, resulting in systemic inflammation during K. pneumoniae infection.

Protective Effect of Gastrodia Elata on Neuronal Cell Damage in Alzheimer's Disease (치매병태(癡呆病態)모델에서 천마(天麻)의 신경세포(神經細胞) 손상(損傷) 보호효과(保護效果))

  • Jung, Young-Su;Kang, Jae-Hyun;Prak, Se-Hwan;Kwon, Young-Mi;Kim, Geun-Woo;Koo, Byung-Soo
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.125-140
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    • 2010
  • Objectives : The purpose of this study is to examine from various angles the protective effect of Gastrodia elata Blume (GEB) against nerve cell death induced by $\beta$-amyloid by using the cell line SH-SY5Y, which is commonly utilized for toxicity testing in nerve cells, and to find out its mechanism of action. Methods : To begin with, as a result of assessing the rate of cell survival by employing MTT reduction assay, the treatment with $\beta$-amyloid at different concentrations caused cytotoxicity, which was inhibited by preprocessing GEB extract. In addition, after $\beta$-amyloid was processed with the cell SH-SY5Y, apoptosis progressed, which was reduced effectively by processing GEB extract. Results : When cytotoxicity was caused by using hydrogen peroxide, a representative ROS, in order to examine the antioxidant effect of GEB, its protective effect was also observed. Apart from ROS, reactive nitrogen species (RNS) are also known to play a crucial role in nerve cell death. The treatment with the NO donor SNAP increased the production of nitric oxide and the expression of iNOS, which was also inhibited by GEB extract. Meanwhile, as an attempt to find out the mechanism of action explaining the antioxidant effect, the intracellular antioxidant enzyme expressions were measured by RT-PCR, which showed that GEB extract increased the expressions of heme oxygenase-1, GAPDH and $\gamma$-glutamate cysteine ligase. Lastly, GEB extract had a protective effect against impaired memory induced by scopolamine in animal models (in vivo). Conclusions : These findings indicate that GEB has a protective effect against the death of cranial nerve cells, suggesting possibilities for the prevention and treatment of AD.

Degradations of human immunoglobulins and hemoglobin by a 60 kDa cysteine proteinase of Trichomonas vaginalis (질편모충의 60 kDa 시스테인 단백분해효소의 인체 면역글로불린 및 헤모글로빈 분해능)

  • Duk-Young MIN;Keun-Hee Hyun;Jae-Sook Ryu;Myoung-Hee AHN;Myung-Hwan CHO
    • Parasites, Hosts and Diseases
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    • v.36 no.4
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    • pp.261-268
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    • 1998
  • The present study was undertaken to investigate the role of cysteine proteinase of Trichomonas vaginalis in escaping from host defense mechanism. A cysteine proteinase of T. vaginalis was purified by affinity chromatography and gel filtration. Optimum pH for the purified proteinase activity was 6.0. The proteinase was inhibited by cysteine and serine proteinase inhibitors such as E-64, NEM, IAA, leupeptin. TPCK and TLCK, and also by $Hg^{2+}$, but not affected by serine-, metallo-, and aspartic proteinase inhibitors such as PMSF, EDTA and pepstatin A. However, it was activated by the cysteine proteinase activator, DTT. The molecular weight of a purified proteinase was 62 kDa on gel filtration and 60 kDa on SDS-PAGE. Interestingly, the purified proteinase was able to degrade serum IgA, secretory IgA, and serum IgG in time- and dose-dependent manners. In addition, the enzyme also degraded hemoglobin in a dose-dependent manner. These results suggest that the acidic cysteine proteinase of T. vaginalis may play a dual role for parasite survival in conferring escape from host humoral defense by degradation of immunoglobulins, and in supplying nutrients to parasites by degradation of hemoglobin.

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Protective effects of PEP-1-Catalase on stress-induced cellular toxicity and MPTP-induced Parkinson's disease

  • Eom, Seon Ae;Kim, Dae Won;Shin, Min Jea;Ahn, Eun Hee;Chung, Seok Young;Sohn, Eun Jeong;Jo, Hyo Sang;Jeon, Su-Jeong;Kim, Duk-Soo;Kwon, Hyeok Yil;Cho, Sung-Woo;Han, Kyu Hyung;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.48 no.7
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    • pp.395-400
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    • 2015
  • Parkinson's disease (PD) is a neurodegenerative disability caused by a decrease of dopaminergic neurons in the substantia nigra (SN). Although the etiology of PD is not clear, oxidative stress is believed to lead to PD. Catalase is antioxidant enzyme which plays an active role in cells as a reactive oxygen species (ROS) scavenger. Thus, we investigated whether PEP-1-Catalase protects against 1-methyl-4-phenylpyridinium (MPP+) induced SH-SY5Y neuronal cell death and in a 1-methyl-4-phenyl-1,2,3,6-trtrahydropyridine (MPTP) induced PD animal model. PEP-1-Catalase transduced into SH-SY5Y cells significantly protecting them against MPP+-induced death by decreasing ROS and regulating cellular survival signals including Akt, Bax, Bcl-2, and p38. Immunohistochemical analysis showed that transduced PEP-1-Catalase markedly protected against neuronal cell death in the SN in the PD animal model. Our results indicate that PEP-1-Catalase may have potential as a therapeutic agent for PD and other oxidative stress related diseases. [BMB Reports 2015; 48(7): 395-400]