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Cytotoxicity of Dichloromethane Extracts of Asian Dust

  • Park, Eun-Jung;Kim, Dae-Seon;Yu, Seong-Do;Park, Kwang-Sik
    • Journal of Environmental Health Sciences
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    • v.36 no.4
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    • pp.271-278
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    • 2010
  • The appearance of Asian Dust (AD) originating from China and Mongolia during spring each year is a meteorological phenomenon periodically observed in extensive regions of East Asia. According to a previous epidemiological study, AD has adverse effects on both human beings and ecosystems. In this study, we collected total suspension particles (TSP) in the AD period and Non-AD (NAD) period. We extracted organic components from TSP using dichloromethane (DCM), and the polyaromatic hydrocarbons (PAHs) were analyzed. The DCM extracts contained PAHs such as benzo(b)fluoranthene, benzo[g,h,i]perylene, benzo(k)fluoranthene, benzo(a)pyrene, and pyrene. No significant difference was observed in cytotoxicity of the DCM extracts from AD versus NAD when tested on the human bronchial epithelial cells, BEAS-2B. e also examined the toxic mechanisms of AD extracts in cultured BEAS-2B cells and RAW264.7 cells, and in BEAS-2B cells observed increased levels of reactive oxygen species (ROS), decreased glutathione (GSH), and induced caspase-3 activity. Increased expression of oxidative stress-related and inflammation- related genes were also observed in BEAS-2B cells, while nitric oxide (NO) levels were increased in RAW264.7 cells. Taken together, the results suggest that in these cultured cells, AD may induce cytotoxicity through oxidative stress and pro-inflammatory signals.

Two-enzyme coupled fluorometric assay of urinary dipeptidase (이원효소 연쇄반응의 형광분석에 의한 Urinary Dipeptidase의 활성도 측정)

  • Park, Haeng Soon;We, Jeoung Soon
    • Analytical Science and Technology
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    • v.8 no.3
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    • pp.359-364
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    • 1995
  • Urinary dipeptidase(Udpase) was assayed by fluorometric analysis of NADH which was produced from an indicator enzyme, L-alanine dehydrogenase. The reaction mixture was consisted of a dipeptide(L-ala-L-ala), ${\beta}-NAD^+$, L-alanine dehydrogenase in 12.5 mM sodium carbonate buffer, pH 9.0, and urinary dipeptidase which initiated the reaction. The fluorescence intensity of NADH was measured as a function of time with the excitation wavelength at 340nm and emission at 460nm. Comparison of this fluorometric method with the conventional spectrophotometric method utilizing glycyldehydrophenylalanine(Gdp) as substrate provided the correlation coefficient of 0.996 and increased the sensitivity more than ten times.

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The conservation of a gilt-bronze Sarira Reliquary, Treasure No. 955 (보물 제955호 선암사 금동팔각원당형사리탑 보존처리)

  • Go, Hyeong-Sun;Yu, Jae-Eun
    • 보존과학연구
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    • s.24
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    • pp.215-227
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    • 2003
  • The gilt-bronze Sarira Reliquary was discovered when repairing three-stories east stone pagoda (Treasure No. 395) at Seungju-eup in Suncheon city in Jeollanam-do Province in August, 1986. Then it was appointed as Treasure in 1988. The Sarira Reliquary had been held in Seonamsa temple, but deterioration on the surface and corrosion had appeared affecting its surface detail. Consequently, the conservation treatment was carried out from November 2002 to March 2003.The corrosion and dirt on the surface of the Sarira Reliquary were cleaned with ethyl alcohol and Benzotriazole was applied to prevent further corrosion. Finally, NAD-10(Paraloid NAD-10), acrylic resin, was used to consolidate the structure. Moreover, after non-destructive analysis to confirm element of alloy, copper, gold, silver and mercury were discovered and this result tells us that it was plated with gold by amalgam. Fibers at the pedestal were examined under the microscope and identified as silk. The total height of this Sarira Reliquary is 6.0cm, the height of lotus pedestal and the roof is 2.7cm and 1.8cm, respectively. The roof and body are joined together, and the lotus pedestal can be separated, on which the octagonal reliquary is impaled. The pedestal consists of 3layers of petals and the surface is decorated with flower pattern. The reliquary is presumed to be created in the 14th century, and it becomes valuable historical material to reveal the secret of metal work in the late Goryeo Dynasty.

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Antibacterial Effect of Amentoflavone and Its Synergistic Effect with Antibiotics

  • Hwang, Ji Hong;Choi, Hyemin;Woo, Eun-Rhan;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.953-958
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    • 2013
  • Selaginella tamariscina is a traditional herb used in medicine. Phytochemical amentoflavone, a biflavonoid class of flavonoids, was isolated from the plant of Selaginella tamariscina. In this study, the antibacterial effects and combination effects of amentoflavone and conventional antibiotics such as ampicillin, cefotaxime, and chloramphenicol were investigated. These results showed that amentoflavone had a considerable antibacterial effect and synergistic interaction with antibiotics against various bacterial strains (fractional inhibitory concentration index ${\leq}$ 0.5), except for Streptococcus mutans. To study the mechanism(s) involved in the synergistic activities between amentoflavone and antibiotics, we detected hydroxyl radical formation using 3'-(p-hydroxyphenyl) fluorescein and measured the $NAD^+/NADH$ ratio by $NAD^+$ cycling assay. The results indicated that the formation of hydroxyl radical would be a cause of the synergistic effect and that this oxidative stress originated from a transient NADH depletion. This study suggests that amentoflavone synergizes with antibiotics and has potential as a therapeutic agent for antimicrobial chemotherapy.

Isolation and Properties of Cytoplasmic α-Glycerol 3-Phosphate Dehydrogenase from the Pectoral Muscle of the Fruit Bat, Eidolon helvum

  • Agboola, Femi Kayode;Thomson, Alan;Afolayan, Adeyinka
    • BMB Reports
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    • v.36 no.2
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    • pp.159-166
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    • 2003
  • Cytoplasmic $\alpha$-glycerol-3-phosphate dehydrogenase from fruit-bat-breast muscle was purified by ion-exchange and affinity chromatography. The specific activity of the purified enzyme was approximately 120 units/mg of protein. The apparent molecular weight of the native enzyme, as determined by gel filtration on Sephadex G-100 was $59,500{\pm}650$ daltons; its subunit size was estimated to be $35,700{\pm}140$ by SDS-polyacrylamide gel electrophoresis. The true Michaelis-Menten constants for all substrates at pH 7.5 were $3.9{\pm}0.7\;mM$, $0.65{\pm}0.05\;mM$, $0.26{\pm}0.06\;mM$, and $0.005{\pm}0.0004\;mM$ for L-glycerol-3-phosphate, $NAD^+$, DHAP, and NADH, respectively. The true Michaelis-Menten constants at pH 10.0 were $2.30{\pm}0.21\;mM$ and $0.20{\pm}0.01\;mM$ for L-glycerol-3-phosphate and $NAD^+$, respectively. The turnover number, $k_{cat}$, of the forward reaction was $1.9{\pm}0.2{\times}10^4\;s^{-1}$. The treatment of the enzyme with 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) under denaturing conditions indicated that there were a total of eight cysteine residues, while only two of these residues were reactive towards DTNB in the native enzyme. The overall results of the in vitro experiments suggest that $\alpha$-glycerol-3-phosphate dehydrogenase of the fruit bat preferentially catalyses the reduction of dihydroxyacetone phosphate to glycerol-3-phosphate.

Dissociation Constant of the Primary Amines and Quaternary Ammonium-methylorange Salts.

  • Kim, Bak-Kwang
    • YAKHAK HOEJI
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    • v.18 no.3
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    • pp.197-202
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    • 1974
  • The data pf dosspcoatopm cpmstamts for aliphatic amines and quaternary ammonium-methylorange salts are based on the electrosatic theory of conductance. Dissociation constants for primary amines nad quaternary ammonium-methylorange salts (1$\times$10$^{-5}$~1$\times$10$^{-3}$ M) in nitrobenzene solution or water solution was evaluated from the relation of the concentration and the electric conduction and the electric conductance at $25^{\circ}C$ Plots of ${\delta}C$ against reciprocal conductance were linear ; hence the center-to-center distance of this salt was 1.75 $\AA$ in nitrobenzene solution.

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Mechanism of Alcoholic Liver Disease

  • 문전옥
    • Journal of Life Science
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    • v.4 no.3
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    • pp.102-112
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    • 1994
  • 알콜성 간장해의 발생, 진전에는 많은 인자가 관여하고 있으며 극히 복잡한 병태를 형성하는데 그 기전으로는 1)간내 [NADH]/[NAD] 비의 상승, 2)에탄올의 주 대사산물인 아세트알데히드에 의한 간장해, 3)면역기구에 의한 간장해, 4)과산화지질, 활성산소 및 free radical 에 의한 장해와 5) 중심정맥역의 hypoxia에 의한 간세포장해 기전을 들 수 있다. 본 총설에서는 알콜을 장기 섭취할 경우 간에서의 대사경로와 현재 고려되고 있는 몇 가지 알콜성 간장해 발생기전에 대한 최근의 연구들을 정리하였다.

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Clusterin(SGP-2) in the Salivary Glands of Insulin Injected Rats under Stress (스트레스에 의한 혈당변화가 타액선내 Clusterin 발현에 미치는 영향)

  • 김선호
    • Journal of Oral Medicine and Pain
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    • v.23 no.4
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    • pp.309-326
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    • 1998
  • In general, the major causative factor of halitosis is thought to be a sulfated compounds. Clusterin, a sulfated glycoprotein-2(SGP-2), is frequently found in diabetic conditions and cold stress conditions. The same result is werum glucose level to diabeteic and cold stress conditions that founded Clusterin. Therefore, this study was performed to examine Clusterin in the slivary glands under stress conditions before insulin injection I.M. Fourty rats were diveded into 3 groups ; 1) 10 rats of gorup I were selected as a control 2) 15 rats beloning to group II were bathed in cold water for 30 seconds twice a day 3) 15 rats in group III received cold stress and injected I.M. with insulin. The rats were sacrifeced at day 0, 3, 5, 7 and 10 of the experiment and the submandibular glands and parotid galnds were removed. RNAs were purified from the salivary of the salivary glands were subjected to Hematoxillin-Eosin stainings and examined under the light microscope. The obtained results were as follows : 1. With immunohistochemistric method, in normal control goup, Clusterin was moderately stained in the intercalated ductal cell of the submandibualr glands, mild stained in the striated ductal cell of the submandibular glands, heavily stained on the cytoplasm of the intercalated ductal cell in the mucous submandibular glands nad slightrly stained in the intercalated ductal cell of the paroted gland, expressed negativity in the acina cell. 2. With immunohistochemistric method, Clusterin slightly increased in the acina cell of the submandibular glands under stress condition at 3 days after experement, moderately stained at 5 days after experiment so revealed positive response. And hearily in the intercalated ductal cell and mildly lin the acina celluar eytoplasm of the parotid glands under stress condition at 3 days experiment. 3. With immunohistochemistric method, no remarkable differences are found between the normal control group and stress conditioned group that insulin administration was performed before. 4. In the stressor-giving group, Clusterin mRNA was porminently expressed in submandibular gland after 5 days after experiment, in parotid gland after 3 days after experiment, performed in immunoelectrophoresis method. 5. In the insulin-injected nad stressor-giving group, Clusterin mRNA was not observed in all experimental submandibular and parotid gland, performed in immunoelectrophoresis method.

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Genetic and Morphologic Identification of Spirometra ranarum in Myanmar

  • Jeon, Hyeong-Kyu;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Sohn, Woon-Mok;Hong, Sung-Jong;Chai, Jong-Yil;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.275-280
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    • 2018
  • In the present study, we identified a Spirometra species of Myanmar origin (plerocercoid) by molecular analysis using mitochondrial cox1 and nad1 genes, as well as by morphological observations of an adult tapeworm. Spargana specimens were collected from a paddy-field in Taik Kyi Township Tarkwa Village, Yangon, Myanmar in December 2017. A total of 5 spargana were obtained from 20 frogs Hoplobatrachus rugulosus; syn: Rana rugulosa (Wiegmann, 1834) or R. tigrina (Steindachner, 1867). The plerocercoids were used for experimental infection of a dog. After 4 weeks of infection, an adult tapeworm was recovered from the intestine of the dog. Morphologically, the distinct features of Spirometra sp. (Myanmar origin) relative to S. erinaceieuropaei and S. decipiens include a uterine morphology comprising posterior uterine coils that larger than the terminal uterine ball and coiling of the uteri diagonally (swirling) rather than spirally. The cox1 sequences (1,566 bp) of the Myanmar-origin Spirometra species showed 97.9% similarity to a reference sequence of S. decipiens (GenBank no. KJ599679) and 90.5% similarity to a reference sequence of S. erinaceieuropaei (GenBank no. KJ599680). Phylogenetic tree topologies were identical and presented high confidence level of values for the 3 major branches of the 3 Spirometra species in cox1 and nad1 genes. These results indicated that Myanmar-origin Spirometra species coincided with those of S. ranarum and may be considered as a valid species.

Quantitative Determination of Total Bile Acids from Bezoar and Bezoar-containing Liquid Preparation by Enzymatic Technique (효소반응법을 이용한 우황 및 우황함유 액상 제제 중 총담즙산의 정량)

  • Ha, In-Sik;Kim, Seung-Hwan;Cha, Bong-Jin;Kwon, Jong-Won;Yang, Joong-Ik;Min, Shin-Hong
    • Journal of Pharmaceutical Investigation
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    • v.21 no.2
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    • pp.67-71
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    • 1991
  • A simple and sensitive method was developed for the quantification of free and conjugated bile acids in bezoar without prior hydrolysis. $3{\alpha}-Hydroxy$ bile acids are first oxidized to 3-keto bile acids in the reaction catalyzed by $3{\alpha}-hydroxysteroid$ $dehydrogenase(3{\alpha}-HSD)$. During this oxidative reaction, an equimolar quantity of nicotinamide adenine dinucleotide(NAD) is reduced to NADH and subsequently oxidized to NAD with concomitant reduction of nitrotetrazolium blue(NTB) to diformazan by the catalytic action of diaphorase. The diformazan has an absorbance maximum at 540 nm. The intensity of the color produced is directly proportional to bile acids concentration in the bezoar extracts. The optimum conditions for the enzymatic reaction such as effects of reaction time, reaction temperature and pH, and stability were investigated. Calibration plots for the sodium chelate observed to be linear and intra-, inter-assay analytical recovery of bile acids averaged $97.65{\pm}3.4%(S.D.)$. Therefore, it is considered that the quality control of total bile acids from bezoar or bezoar-containing liquid preparation using this simple and sensitive assay system will be acceptable. Also current bezoars and bezoar-containing liauid preparations were examined their total bile acids from this method.

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