• Title/Summary/Keyword: in vitro metabolite

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New Yeast Cell-Based Assay System for Screening Histone Deacetylase 1 Complex Disruptor

  • Jeon, Kwon-Ho;Kim, Min-Jung;Kim, Seung-Young
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.286-291
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    • 2002
  • Histone deacetylase I (HDAC1) works as one of the components in a nucleosome remodeling (NuRD) complex that consists of several proteins, including metastasis-associated protein 1 (MTA1). Since the protein-protein interaction of HDAC1 and MTA1 would appear to be important for both the integrity and functionality of the HDAC1 complex, the interruption of the HDAC1 and MTA1 interaction may be an efficient way to regulate the biological function of the HDAC1 complex. Based on this idea, a yeast two-hybrid system was constructed with HDAC1 and MTA1 expressing vectors in the DNA binding and activation domains, respectively. To verify the efficiency of the assay system, 3,500 microbial metabolite libraries were tested using the paper disc method, and KB0699 was found to inhibit the HDAC1 and MTA1 interaction without any toxicity to the wild-type yeast. Furthermore, KB0699 blocked the interaction of HDAC1 and MTA1 in an in vitro GST pull down assay and induced morphological changes in B16/BL6 melanoma cells, indicating the interruption of the HDAC1 complex function. Accordingly, these results demonstrated that the yeast assay strain developed in this study could be a valuable tool for the isolation of a HDAC1 complex disruptor.

Protective effect of Korean Red Ginseng against chemotherapeutic drug-induced premature catagen development assessed with human hair follicle organ culture model

  • Keum, Dong In;Pi, Long-Quan;Hwang, Sungjoo Tommy;Lee, Won-Soo
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.169-175
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    • 2016
  • Background: Chemotherapy-induced alopecia (CIA) is one of the most distressing side effects for patients undergoing chemotherapy. This study evaluated the protective effect of Korean Red Ginseng (KRG) on CIA in a well-established in vitro human hair follicle organ culture model as it occurs in vivo. Methods: We examined whether KRG can prevent premature hair follicle dystrophy in a human hair follicle organ culture model during treatment with a key cyclophosphamide metabolite, 4-hydroperoxycyclophosphamide (4-HC). Results: 4-HC inhibited human hair growth, induced premature catagen development, and inhibited proliferation and stimulated apoptosis of hair matrix keratinocytes. In addition, 4-HC increased p53 and Bax protein expression and decreased Bcl2 protein expression. Pretreatment with KRG protected against 4-HC-induced hair growth inhibition and premature catagen development. KRG also suppressed 4-HC-induced inhibition of matrix keratinocyte proliferation and stimulation of matrix keratinocyte apoptosis. Moreover, KRG restored 4-HC-induced p53 and Bax/Bcl2 expression. Conclusion: Overall, our results indicate that KRG may protect against 4-HC-induced premature catagen development through modulation of p53 and Bax/Bcl2 expression.

Antinociceptive Effects of Prim-O-Glucosylcimifugin in Inflammatory Nociception via Reducing Spinal COX-2

  • Wu, Liu-Qing;Li, Yu;Li, Yuan-Yan;Xu, Shi-hao;Yang, Zong-Yong;Lin, Zheng;Li, Jun
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.418-425
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    • 2016
  • We measured anti-nociceptive activity of prim-o-glucosylcimifugin (POG), a molecule from Saposhnikovia divaricate (Turcz) Schischk. Anti-nociceptive or anti-inflammatory effects of POG on a formalin-induced tonic nociceptive response and a complete Freund's adjuvant (CFA) inoculation-induced rat arthritis pain model were studied. Single subcutaneous injections of POG produced potent anti-nociception in both models that was comparable to indomethacin analgesia. Anti-nociceptive activity of POG was dose-dependent, maximally reducing pain 56.6% with an $ED_{50}$ of 1.6 mg. Rats given POG over time did not develop tolerance. POG also time-dependently reduced serum TNF${\alpha}$, IL-$1{\beta}$ and IL-6 in arthritic rats and both POG and indomethacin reduced spinal prostaglandin E2 ($PGE_2$). Like indomethacin which inhibits cyclooxygenase-2 (COX-2) activity, POG dose-dependently decreased spinal COX-2 content in arthritic rats. Additionally, POG, and its metabolite cimifugin, downregulated COX-2 expression in vitro. Thus, POG produced potent anti-nociception by downregulating spinal COX-2 expression.

Improvement in Clonal Propagation of Hemidesmus indicus R. Br. through Adenine Sulphate

  • Misra Neeta;Misra Pratibha;Datta S.K.;Mehrotra Shanta
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.239-244
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    • 2003
  • A protocol has been developed for rapid large scale clonal propagation of an aromatic endangered medicinal plant, Hemidesmus indicus R. Br. with the elimination of the problems such as premature leaf fall and callus formation during caulogenesis and rhizogenesis. Multiple shoots were induced from shoot tip and nodal explants on Murashige and Skoog (MS) medium supplemented with 1 mg/L Benzylaminopurine (BAP) and 0.5 mg/L Napthaleneaceticacid (NAA). Addition of 15 mg/L adenine sulphate to the above medium checked leaf abscission completely, reduced the time required for caulogenesis and restored morphogenetic potential after several subcultures. The in vitro grown propagules were rooted in 1/2 MS medium supplemented with 2 mg/L Indolebutyric acid (IBA) +1 mg/L NAA and sucrose 0.7% (w/v). Addition of charcoal at 100 mg/L to the rooting medium quickened root initiation with a complete check on callus formation. The effect of sucrose concentration on both caulogenesis and rhizogenesis was also studied. The resultant plantlets were acclimatized and grown in fields where ninety eight percent of the rooted shoots survived and grew normally. The estimation of the secondary metabolite content in the shoots of the regenerated plant and the mother plant indicated that the concentration of the three secondary metabolites lupeol, vanillin and rutin was similar.

Methylated Organic Metabolites of Arsenic and their Cardiovascular Toxicities

  • Bae, Ok-Nam;Lim, Kyung-Min;Noh, Ji-Yoon;Kim, Keun-Young;Lim, Eun-Kyung;Chung, Jin-Ho
    • Toxicological Research
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    • v.24 no.3
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    • pp.161-167
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    • 2008
  • Recently, arsenic-toxicity has become the major focus of strenuous assessment and dynamic research from the academy and regulatory agency. To elucidate the cause and the mechanism underlying the serious adverse health effects from chronic ingestion of arsenic-contaminated drinking water, numerous studies have been directed on the investigation of arsenic-toxicity using various in vitro as well as in vivo systems. Neverthless, some questions for arsenic effects remain unexplained, reflecting the contribution of unknown factors to the manifestation of arsenic-toxicity. Interestingly, very recent studies on arsenic metabolites have discovered that trivalent methylated arsenicals show stronger cytotoxic and genotoxic potentials than inorganic arsenic or pentavalent metabolites, arguing that these metabolites could play a key role in arsenic-associated disorders. In this review, recent progress and literatures are summarized on the metabolism of trivalent methylated metabolites and their toxicity on body systems including cardiovascular system in an effort to provide an insight into the future research on arsenic-associated disorders.

Recent Research Trends of Sayeok-tang Used in Ischemic Heart Disease (허혈성 심질환에 활용된 사역탕(四逆湯)의 최신 연구 동향)

  • Jang, Il-Woong;Hong, Jun-yeong;Lee, Soong-in
    • 대한상한금궤의학회지
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    • v.13 no.1
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    • pp.1-19
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    • 2021
  • Purpose : Sayeok-tang (Sini decoction, SND) is a cold-dispelling formula used for cold deficiency syndrome and is composed of Aconiti Lateralis Radix Preperata, Zingiberis Rhizoma, and Glycyrrhizae Radix et Rhizoma. It is used for diseases such as myocardial infarction, heart failure, acute and chronic gastroenteritis, and gastric effusion. This study proposes the possibility of expansion of basic research and clinical applications for ischemic heart disease (IHD) through systematic analysis of domestic and foreign studies on SND. Methods : We collected studies within the last 10 years on the use of SND in IHD and excluded those lacking relevance. Selected studies were classified by research method and the main themes of the studies were analyzed for each classification. Result: Out of 15 studies, there were 5 animal studies, 8 metabolite analyses in animals, 1 in vitro study, and 1 systematic review. Our review suggests that SND may be used as an adjuvant to nitroglycerin and percutaneous transluminal coronary angioplasty, and may improve symptoms and quality of life of patients with IHD. Myocardial protective effects through antioxidant, anti-inflammatory, anti-apoptotic, and anti-hypertensive actions were confirmed through these studies. Effects on carbohydrate, protein, and lipid metabolism were also reported. Conclusions : This study suggests that SND has potential as a treatment for IHD.

Antimicrobial active clones from soil metagenomic library

  • H. K. Lim;Lee, E. H;Kim, J.C.;Park, G. J.;K S. Jang;Park, Y. H.;K Y. Cho;S, W. Lee
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.108.1-108
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    • 2003
  • Soil metagenome is untapped total microbial genome including that of the majority of unculturable bacteria present in soil. We constructed soil metagenomic library in Escherichia coli using DNA directly extracted from two different soils, pine tree rhizosphere soil and forest topsoil. Metagenomic libraries constructed from pine tree rhizosphere soil and forest topsoil consisted of approximately 33,700 clones and 112,000 clones with average insert DNA size of 35-kb, respectively. Subsequently, we screened the libraries to select clones with antimicrobial activities against Saccharomyces cerevisiae and Agrobacterium tumefaciens using double agar layer method. So far, we have a clone active against S. cerevisiae and a clone active against A. tumefaciens from the forest topsoil library. In vitro mutagenesis and DNA sequence analysis of the antifungal clone revealed the genes involved in the biosynthesis of antimicrobial secondary metabolite. Metagenomic libraries constructed in this study would be subject to search for diverse genetic resources related with useful microbial products.

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The Lipogenic Capacity of Hepatocytes and Lipolytic Rate of Adipocytes in Tsaiya Ducks during Growing and Laying Periods

  • Lien, T.F.;Jan, D.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.8
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    • pp.1258-1262
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    • 1999
  • With an attempt to elucidating the lipid metabolism of Tsaiya ducks, thirty ducks at growing (8 weeks of age) and laying periods (10 weeks after the onset of laying) were examined, respectively. The ducks were randomly allocated into ad libitum feeding and 3-day fasting groups, to investigate their in vitro hepatocytes lipogenesis capacity and adipocytes lipolysis rate. Results indicate that (1) the capacity of hepatocytes incorporation of glucose and acetate into total lipid and metabolite of $^{14}CO_2$ production during the laying period was greater than during the growing period. Approximately 50% of the glucose or acetate converted into triacylglycerol (TG) by the hepatocytes were recovered as fatty acid during the growing period, while it was 65-70% during the laying period. (2) Acetate used for lipogenesis ability was superior to glucose in both periods. (3) The adipocytes lipolysis rate was increased significantly (p<0.05) by fasting. In contrast, the capacity of incorporated glucose or acetate into total lipid, triacylglycerol, fatty acid and glycerol by hepatocytes was reduced significantly (p<0.05) by fasting.

The Acetylcholinesterase Inhibitory Activity of the EtOH Extract of Chaenomelis Fructus and its effects on the Metabolism of Amyloid Precursor Protein in Neuroblastoma Cells (모과 에탄올 추출물의 아세틸콜린에스테라제 저해활성과 신경세포에서 아밀로이드 전구단백질의 대사에 미치는 영향)

  • Kim, Ju Eun;Jo, Youn Jeong;Leem, Jae Yoon
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.327-333
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    • 2015
  • Alzheimer's disease (AD) is a progressive neurodegenerative disorder symptomatically characterized by impairment in memory and cognitive abilities. AD is characterized pathologically by the deposition of ${\beta}$-amyloid ($A{\beta}$) peptides of 40-42 residues, which are generated by processing of amyloid precursor protein (APP). $A{\beta}$ has been believed to be neurotoxic and now is also considered to have a role on the mechanism of memory dysfunction. In this study, we tested that EtOH extract of the fruits of Chaenomeles sinensis Koehne (CSE) affects on the processing of APP from the APPswe over-expressing Neuro2a cell line. We found that CSE increased over 2 folds of the $sAPP{\alpha}$ secretion level, a metabolite of ${\alpha}$-secretase. We showed that CSE reduced the secretion level of $A{\beta}42$ and $A{\beta}40$ by down regulation of ${\beta}$-secretase (BACE) without cytotoxicity. Furthermore, we found that CSE inhibited BACE and acetylcholinesterase activity in vitro. We suggest that Chaenomelis Fructus may be an useful source to develop a herbal medicine for AD.

Study in vivo metabolism using nuclear medical examination of the dose rate reduction (체내대사를 이용한 핵의학적 검사의 선량률 저감에 관한 연구)

  • Kang, Yong-Gil;Na, Soo-Kyung;Hong, Jin-Woong;Lee, Gui-Won;Kim, Nak-Sang
    • Korean Journal of Digital Imaging in Medicine
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    • v.14 no.2
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    • pp.75-81
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    • 2012
  • The purpose of this study was administered to the body for examination and treatment, high-energy radioactive isotope(F-18, I-131) in vitro discharge experiments. Increasing exposed dose of radiation to health professionals is caused by the increase of PET/CT use and a radioactive isotope. Therefore, the high-energy isotope F-18 and I-131 after administration about using Metabolite excretion was studied. As a results of this study, patients had plenty of fluids for testing and treatment alone administered radiopharmaceuticals can be more than twice as fast excretion induced emissions. Also was able to get a better image space to reduce the dose rate.

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