• Title/Summary/Keyword: Mechanism synthesis

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CD38 Inhibition Protects Fructose-Induced Toxicity in Primary Hepatocytes

  • Soo-Jin Lee;Sung-E Choi;Seokho Park;Yoonjung Hwang;Youngho Son;Yup Kang
    • Molecules and Cells
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    • v.46 no.8
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    • pp.496-512
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    • 2023
  • A fructose-enriched diet is thought to contribute to hepatic injury in developing non-alcoholic steatohepatitis (NASH). However, the cellular mechanism of fructose-induced hepatic damage remains poorly understood. This study aimed to determine whether fructose induces cell death in primary hepatocytes, and if so, to establish the underlying cellular mechanisms. Our results revealed that treatment with high fructose concentrations for 48 h induced mitochondria-mediated apoptotic death in mouse primary hepatocytes (MPHs). Endoplasmic reticulum stress responses were involved in fructose-induced death as the levels of phosho-eIF2α, phospho-C-Jun-N-terminal kinase (JNK), and C/EBP homologous protein (CHOP) increased, and a chemical chaperone tauroursodeoxycholic acid (TUDCA) prevented cell death. The impaired oxidation metabolism of fatty acids was also possibly involved in the fructose-induced toxicity as treatment with an AMP-activated kinase (AMPK) activator and a PPAR-α agonist significantly protected against fructose-induced death, while carnitine palmitoyl transferase I inhibitor exacerbated the toxicity. However, uric acid-mediated toxicity was not involved in fructose-induced death as uric acid was not toxic to MPHs, and the inhibition of xanthine oxidase (a key enzyme in uric acid synthesis) did not affect cell death. On the other hand, treatment with inhibitors of the nicotinamide adenine dinucleotide (NAD)+-consuming enzyme CD38 or CD38 gene knockdown significantly protected against fructose-induced toxicity in MPHs, and fructose treatment increased CD38 levels. These data suggest that CD38 upregulation plays a role in hepatic injury in the fructose-enriched diet-mediated NASH. Thus, CD38 inhibition may be a promising therapeutic strategy to prevent fructose-enriched diet-mediated NASH.

Effect of Erythritol on Glucosyl Transferase and Fructosyl Transferase Gene Expression in Streptococcus mutans (Streptococcus mutans의 Glucosyl Transferase와 Fructosyl Transferase 유전자 발현에 대한 Erythritol의 효과)

  • Young-Nam PARK;Jae-Ki RYU
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.3
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    • pp.151-158
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    • 2023
  • Erythritol is a sweetener produced by yeast from glucose and a natural sugar found in fermented foods such as mushrooms, wine, fruits, rice wine, and soy sauce. Correct information and basic data when producing or using products for preventing dental caries by checking the gene expression patterns of glucosyl transferase (GTF) and fructosyl transferase (FTF) of Streptococcus mutans in erythritol and other sweeteners it was implemented to provide. Erythritol inhibited the growth of Streptococcus mutans, which is involved in dental caries. When used as a sweetener to replace sucrose, erythritol had an excellent caries-preventative effect. In particular, erythritol reduced the expressions of gtfB, gtfC, gtfD, and FTF, which are related to the synthesis of extracellular polysaccharides, and thereby reduced the formation of dental plaque and the attachment rate of bacteria to tooth surfaces. The study shows erythritol has potential use as an anticariogenic sweetener that inhibits the mechanism underlying caries caused by Streptococci.

The molecular mechanism of propionate-regulating gluconeogenesis in bovine hepatocytes

  • Rui Pang;Xiao Xiao;Tiantian Mao;Jiajia Yu;Li Huang;Wei Xu;Yu Li;Wen Zhu
    • Animal Bioscience
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    • v.36 no.11
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    • pp.1693-1699
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    • 2023
  • Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

Physiological and transcriptome analysis of acclimatory response to cold stress in marine red alga Pyropia yezoensis

  • Li-Hong Ma;Lin Tian;Yu-Qing Wang;Cong-Ying Xie;Guo-Ying Du
    • ALGAE
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    • v.39 no.1
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    • pp.17-30
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    • 2024
  • Red macroalga Pyropia yezoensis is a high valuable cultivated marine crop. Its acclimation to cold stress is especially important for long cultivation period across winter in coasts of warm temperate zone in East Asia. In this study, the response of P. yezoensis thalli to low temperature was analyzed on physiology and transcriptome level, to explore its acclimation mechanism to cold stress. The results showed that the practical photosynthesis activity (indicated by ΦPSII and qP) was depressed and pigment allophycocyanin content was decreased during the cold stress of 48 h. However, the Fv/Fm and non-photochemical quenching increased significantly after 24 h, and the average growth rate of thalli also rebounded from 24 to 48 h, indicating a certain extent of acclimation to cold stress. On transcriptionally, the low temperature promoted the expression of differentially expressed genes (DEGs) related to carbohydrate metabolism and energy metabolism, while genes related to photosynthetic system were depressed. The increased expression of DEGs involved in ribosomal biogenesis and lipid metabolism which could accelerate protein synthesis and enhance the degree of fatty acid unsaturation, might help P. yezoensis thallus cells to cope with cold stress. Further co-expression network analysis revealed differential expression trends along with stress time, and corresponding hub genes play important roles in the systemic acquired acclimation to cold stress. This study provides basic mechanisms of P. yezoensis acclimation to cold temperature and may aid in exploration of functional genes for genetic breeding of economic macroalgae.

Molybdenum-Based Electrocatalysts for Direct Alcohol Fuel Cells: A Critical Review

  • Gaurav Kumar Yogesh;Rungsima Yeetsorn;Waritnan Wanchan;Michael Fowler;Kamlesh Yadav;Pankaj Koinkar
    • Journal of Electrochemical Science and Technology
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    • v.15 no.1
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    • pp.67-95
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    • 2024
  • Direct alcohol fuel cells (DAFCs) have gained much attention as promising energy conversion devices due to their ability to utilize alcohol as a fuel source. In this regard, Molybdenum-based electrocatalysts (Mo-ECs) have emerged as a substitution for expensive Pt and Ru-based co-catalyst electrode materials in DAFCs, owing to their unique electrochemical properties useful for alcohol oxidation. The catalytic activity of Mo-ECs displays an increase in alcohol oxidation current density by several folds to 1000-2000 mA mgPt-1, compared to commercial Pt and PtRu catalysts of 10-100 mA mgPt-1. In addition, the methanol oxidation peak and onset potential have been significantly reduced by 100-200 mV and 0.5-0.6 V, respectively. The performance of Mo-ECs in both acidic and alkaline media has shown the potential to significantly reduce the Pt loading. This review aims to provide a comprehensive overview of the bifunctional mechanism involved in the oxidation of alcohols and factors affecting the electrocatalytic oxidation of alcohol, such as synthesis method, structural properties, and catalytic support materials. Furthermore, the challenges and prospects of Mo-ECs for DAFCs anode materials are discussed. This in-depth review serves as valuable insight toward enhancing the performance and efficiency of DAFC by employing Mo-ECs.

The Anti-Diabetic Pinitol Improves Damaged Fibroblasts

  • Ji-Yong Jung;Joong Hyun Shim;Su Hae Cho;Il-Hong Bae;Seung Ha Yang;Jinsick Kim;Hye Won Lim;Dong Wook Shin
    • Biomolecules & Therapeutics
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    • v.32 no.2
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    • pp.224-230
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    • 2024
  • Pinitol (3-O-Methyl-D-chiro-inositol) has been reported to possess insulin-like effects and is known as one of the anti-diabetic agents to improve muscle, liver, and endothelial cells. However, the beneficial effects of pinitol on the skin are not well known. Here, we investigated whether pinitol had effects on human dermal fibroblasts (HDFs), and human dermal equivalents (HDEs) irradiated with ultraviolet A (UVA), which causes various damages including photodamage in the skin. We observed that pinitol enhanced wound healing in UVA-damaged HDFs. We also found that pinitol significantly antagonized the UVA-induced up-regulation of matrix metalloproteinase 1 (MMP1), and the UVA-induced down-regulation of collagen type I and tissue inhibitor of metalloproteinases 1 (TIMP1) in HDEs. Electron microscopy analysis also revealed that pinitol remarkably increased the number of collagen fibrils with regular banding patterns in the dermis of UVA-irradiated human skin equivalents. Pinitol significantly reversed the UVA-induced phosphorylation levels of ERK and JNK but not p38, suggesting that this regulation may be the mechanism underlying the pinitol-mediated effects on UVA-irradiated HDEs. We also observed that pinitol specifically increased Smad3 phosphorylation, which is representative of the TGF-β signaling pathway for collagen synthesis. These data suggest that pinitol exerts several beneficial effects on UVA-induced damaged skin and can be used as a therapeutic agent to improve skin-related diseases.

The Analysis of Expression of Autoinducer Synthesis Genes Involved in Quorum Sensing among Catheter Associated Bacteria (요로감염에 관여하는 카테터 내 박테리아의 Quorum Sensing 관련 autoinducer 합성 유전자의 발현분석)

  • Lee, Mi-Hye;Seo, Pil-Soo;Lee, Ji-Youl;Peck, Kyong-Ran;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.277-285
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    • 2006
  • The most biofilm forming bacteria in catheter, Esctherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were isolated and identified from a patient's catheter occuring catheter-associated urinary tract infection (CA-UTI). We examined mRNA expression and its quantification of AIs synthetic genes encoding signal substance of quorum sensing from each bacterial species in order to elucidated quorum sensing mechanism. Both pure cultures for each bacterial strains and a mixed cultures with three were grown for 24 hr and 30 days. Initial densities to be able to detect mRNA expression oil single strains culture were shown at $2.4{\times}10^5$ CFU/ml, $5.4{\times}10^6$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $6.9{\times}10^4$ CFU/ml of P. aeruginosa for rhlI and lasI. Also, in mixed culture of three, initial cell densities of mRNA expression were appear to at $7.3{\times}10^5$ CFU/ml, $1.6{\times}10^7$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $2.1{\times}10^5$ CFU/ml of P. aeruginosa for rhlI and lasI. Each AIs synthetic gene was expressed in initial cell density and the mRNA expression of the genes were detected continously during 30 days. And then, the quantification of mRNA expression level of ygaG, rhlI, last, and luxS which were related AIs synthesis was done each time point by real-time RT-PCR. Interestingly, the mRNA levels of ygaG, rhlI, lasI, and luxS from the mixed culture was higher than those from each single strain culture. In the case of E. coli ygaG, the amount of transcript from the mixed culture was at least 30 times for that from single culture. In the case of P. aeruginosa rhlI and lasI, the amount of transcript from the mixed culture was at least 40 times and 250 times for that from single strain culture. In the case of S. aureus luxS, the amount of transcript from the mixed culture was at least 5 times for that from single strain culture. And specially, the mRNA expression of rhlI and lasI of P. aeruginosa showed the highest efficency among four AIs synthetic genes.

Signal Transduction of MUC5AC Expression in Airway Mucus Hypersecretory Disease (기도의 점액 과분비 질환에서 MUC5AC의 발현의 신호 전달 경로에 관한 연구)

  • Shim, Jae Jeong
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.21-30
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    • 2003
  • Background : Mucin synthesis in airways has been reported to be regulated by the epidermal growth factor receptor (EGFR) system. Epidermal growth factor receptor transactivation was identified as a critical element in G-protein-coupled receptors (GPCRs)-induced mitogenic signaling. EGF receptor transactivation by G-protein-coupled receptors requires metalloproteinase cleavage of proHB-EGF. This study was hypothesized that lipopolysaccharide (LPS)-induced mucin production associates with epidermal growth factor receptor transactivation, and MUC5AC production associates with epidermal growth factor receptor transactivation by G-protein-coupled receptors that regulates by metalloproteinase. Method : MUC5AC mucin production was examined in NCI-H292 cells and MUC5AC protein synthesis was assessed using ELISA. For the evaluation of mechanism of LPS-induced MUC5AC production, $TNF{\alpha}$ was measured using ELISA with or without pretreatment of heterotrimeric G-protein inhibitor, mastoparan. MUC5AC protein was measure with pretreatment of polyclonal $TNF{\alpha}$ antibody or mastoparan on LPS-induced MUC5AC production. For the evaluation of relation of G-protein and MUC5AC production, G-protein stimulant, mastopara-7, or matrix metalloproteinase, ADAM10, was added to NCI-H292 cells. MUC5AC protein was measure with pretreatment of polyclonal EGF antibody on mastoparan-7-induced MUC5AC production. Results : LPS alone did not increase significantly MUC5AC production. LPS with $TNF{\alpha}$ induced dose-dependently MUC5AC production in NCI-H292 cells. LPS increased dose-dependently $TNF{\alpha}$ secretion, which was inhibited by mastoparan. LPS with $TNF{\alpha}$-induced MUC5AC production was inhibited by neutralizing polyclonal $TNF{\alpha}$ antibody, mastoparan or AG 1472. Mastoparan-7 or ADAM10 increased dose-dependently MUC5AC production, which was inhibited by polyclonal neutralizing EGF antibody. Conclusion : In LPS-induced MUC5AC synthesis, LPS causes $TNF{\alpha}$ secretion, which induces EGFR expression. EGFR tyrosine kinase phosphorylation result in MUC5AC production. EGF-R transactivation by G-protein-coupled receptors requires matrix metalloproteinase cleavage of proHB-EGF.

Uptake of Butachlor by Rice Seedlings and Its Phytotoxic Action to the Physiological Activities (수도묘(水稻苗)의 Butachlor 흡수(吸收) 및 약해발생(藥害發生) 특성(特性)에 관한 생리적(生理的) 연구(硏究))

  • Chung, Bong-Jin;Kwon, Yong-Woong
    • Korean Journal of Weed Science
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    • v.1 no.1
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    • pp.57-68
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    • 1981
  • To clarify the mode of uptake of butachlor (2-chloro-2', 6'-diethyl-N-(butoxymethyl) acetanilide) by rice seedlings, its phytotoxic action to growth and physiological activities, studies were conducted with rice seedlings, at the 6th or 7th leaf-stage, which were treated with nutrient solution containing butachlor 0, 1.8, 3.6, 7.2, 10.8 or 14.4 ppm for 1, 2 or 4 days, in other case, the solutions were thereafter renewed with the untreated nutrient solution for further growth. Uptake of butachlor by rice seedlings increased linearly with increase of its concentration and duration of uptake. Butachlor inhibited root growth more than shoot growth, furthermore, the inhibitory effect on the shoot growth was greater in height than in weight or leafing rate. After 4 day-treatment, the rates of shoot growth in weight were delayed for 4 days. Butachlor inhibited water uptake rapidly and linearly with increase of its external concentration. The reduced uptake of water was followed by slow increase in the stomatal resistance of leaves. Upon completion of butachlor treatment, rate of water uptake was recovered rapidly, but the stomatal resistance with lag in time. Butachlor did not affect the uptake of cation such as ammonium, potassium and calcium, but inhibited substantially uptake of nitrate in proportion to its concentration. Especially, butachlor did not affect synthesis and degradation of nitrate reductase. In addition, butachlor has shown much greater binding to the lipidic substances from rice roots than the proteinous material. The primary mechanism of phytotoxic action of butachlor does not seem to be its effect on the protein synthesis, but great affinity to membranes. The inhibition of water uptake, and its subsequent closure of stomates is thought very important for reduced growth under mild phytotoxicity.

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Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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