<b>Objective:</b> Evaluation antiviral effects of <i>Withania somnifera</i> (Ashwagandha) leaf extract against HCV. <b>Methods:</b> cell proliferation was assessed using MTT assay a...<b>Objective:</b> Evaluation antiviral effects of <i>Withania somnifera</i> (Ashwagandha) leaf extract against HCV. <b>Methods:</b> cell proliferation was assessed using MTT assay after isolation of lymphocyte cells and treated with Ashwagandha water extract (ASH-WX) (6.25 mg/ml - 100 mg/ml). Assessment of quantitative Real-time PCR, Colony forming assay, TNF-<i>α</i> and molecular docking studies after infection of normal lymphocyte cells with 1 ml (1.5 × 10<sup>6</sup> HCV) serum then incubated with ASH-WX at concentration 25 mg/ml & 50 mg/ml. <b>Results:</b> MTT assay revealed a significant increase (p < 0.001) in normal lymphocyte proliferation at all concentration’s particularity at 25 mg/ml with SI (6.06) and at 50 mg/ml with (5.8). While TNF-<i>α</i> significantly decreased following ASH-WX treatment compared with control untreated infected cells (p < 0.05). PCR results showed a marked viral load reduction after treatment by ASH-WX at concentration 25 mg/ml to 6.241 × 10<sup>3</sup> IU/mL. Colony formation assay test revealed colony formation reduction compared to positive untreated control. Molecular docking analysis revealed good prediction of binding between Ashwagandha and NS5B and PKN2 compared to Sovaldi. <b>Conclusion: </b> ASH-WX may be a powerful antiviral against HCV infection.展开更多
Pancreatic cancer(Pa C) shows a clear tendency to increase in the next years and therefore represents an important health and social challenge. Currently, there is an important need to find biomarkers for PaC early de...Pancreatic cancer(Pa C) shows a clear tendency to increase in the next years and therefore represents an important health and social challenge. Currently, there is an important need to find biomarkers for PaC early detection because the existing ones are not useful for that purpose. Recent studies have indicated that there is a large window of time for PaC early detection, which opens the possibility to find early biomarkers that could greatly improve the dismal prognosis of this tumor. The present manuscript reviews the state of the art of the existing PaC biomarkers. It focuses on the anomalous glycosylation process and its role in PaC. Glycan structures of glycoconjugates such as glycoproteins are modified in tumors and these modifications can be detected in biological fluids of the cancer patients. Several studies have found serum glycoproteins with altered glycan chains in PaC patients, but they have not shown enough specificity for PaC. To find more specific cancer glycoproteins we propose to analyze the glycan moieties of a battery of glycoproteins that have been reported to increase in PaC tissues and that can also be found in serum. The combination of these new candidate glycoproteins with their aberrant glycosylation together with the existing biomarkers could result in a panel, which would expect to give better results as a new tool for early diagnosis of PaC and to monitor the disease.展开更多
The SMYD(SET and MYND domain)family of lysine methyltransferases(KMTs)plays pivotal roles in various cellular processes,including gene expression regulation and DNA damage response.Initially identified as genuine hist...The SMYD(SET and MYND domain)family of lysine methyltransferases(KMTs)plays pivotal roles in various cellular processes,including gene expression regulation and DNA damage response.Initially identified as genuine histone methyltransferases,specific members of this family have recently been shown to methylate non-histone proteins such as p53,VEGFR,and the retinoblastoma tumor suppressor(pRb).To gain further functional insights into this family of KMTs,we generated the protein interaction network for three different human SMYD proteins(SMYD2,SMYD3,and SMYD5).Characterization of each SMYD protein network revealed that they associate with both shared and unique sets of proteins.Among those,we found that HSP90 and several of its co-chaperones interact specifically with the tetratrico peptide repeat(TPR)-containing SMYD2 and SMYD3.Moreover,using proteomic and biochemical techniques,we provide evidence that SMYD2 methylates K209 and K615 on HSP90 nucleotide-binding and dimerization domains,respectively.In addition,we found that each methylation site displays unique reactivity in regard to the presence of HSP90 co-chaperones,pH,and demethylation by the lysine amine oxidase LSD1,suggesting that alternative mechanisms control HSP90 methylation by SMYD2.Altogether,this study highlights the ability of SMYD proteins to form unique protein complexes that may underlie their various biological functions and the SMYD2-mediated methylation of the key molecular chaperone HSP90.展开更多
文摘<b>Objective:</b> Evaluation antiviral effects of <i>Withania somnifera</i> (Ashwagandha) leaf extract against HCV. <b>Methods:</b> cell proliferation was assessed using MTT assay after isolation of lymphocyte cells and treated with Ashwagandha water extract (ASH-WX) (6.25 mg/ml - 100 mg/ml). Assessment of quantitative Real-time PCR, Colony forming assay, TNF-<i>α</i> and molecular docking studies after infection of normal lymphocyte cells with 1 ml (1.5 × 10<sup>6</sup> HCV) serum then incubated with ASH-WX at concentration 25 mg/ml & 50 mg/ml. <b>Results:</b> MTT assay revealed a significant increase (p < 0.001) in normal lymphocyte proliferation at all concentration’s particularity at 25 mg/ml with SI (6.06) and at 50 mg/ml with (5.8). While TNF-<i>α</i> significantly decreased following ASH-WX treatment compared with control untreated infected cells (p < 0.05). PCR results showed a marked viral load reduction after treatment by ASH-WX at concentration 25 mg/ml to 6.241 × 10<sup>3</sup> IU/mL. Colony formation assay test revealed colony formation reduction compared to positive untreated control. Molecular docking analysis revealed good prediction of binding between Ashwagandha and NS5B and PKN2 compared to Sovaldi. <b>Conclusion: </b> ASH-WX may be a powerful antiviral against HCV infection.
基金Supported by the Spanish Ministry of Economy and Competitivity,No.BIO 2015-66356-Rthe Generalitat de Catalunya Grant,No.2017SGR673
文摘Pancreatic cancer(Pa C) shows a clear tendency to increase in the next years and therefore represents an important health and social challenge. Currently, there is an important need to find biomarkers for PaC early detection because the existing ones are not useful for that purpose. Recent studies have indicated that there is a large window of time for PaC early detection, which opens the possibility to find early biomarkers that could greatly improve the dismal prognosis of this tumor. The present manuscript reviews the state of the art of the existing PaC biomarkers. It focuses on the anomalous glycosylation process and its role in PaC. Glycan structures of glycoconjugates such as glycoproteins are modified in tumors and these modifications can be detected in biological fluids of the cancer patients. Several studies have found serum glycoproteins with altered glycan chains in PaC patients, but they have not shown enough specificity for PaC. To find more specific cancer glycoproteins we propose to analyze the glycan moieties of a battery of glycoproteins that have been reported to increase in PaC tissues and that can also be found in serum. The combination of these new candidate glycoproteins with their aberrant glycosylation together with the existing biomarkers could result in a panel, which would expect to give better results as a new tool for early diagnosis of PaC and to monitor the disease.
基金supported by a Canadian Institutes for Health Research grant (GMX-209406 to J.-F.C.and D.F.).
文摘The SMYD(SET and MYND domain)family of lysine methyltransferases(KMTs)plays pivotal roles in various cellular processes,including gene expression regulation and DNA damage response.Initially identified as genuine histone methyltransferases,specific members of this family have recently been shown to methylate non-histone proteins such as p53,VEGFR,and the retinoblastoma tumor suppressor(pRb).To gain further functional insights into this family of KMTs,we generated the protein interaction network for three different human SMYD proteins(SMYD2,SMYD3,and SMYD5).Characterization of each SMYD protein network revealed that they associate with both shared and unique sets of proteins.Among those,we found that HSP90 and several of its co-chaperones interact specifically with the tetratrico peptide repeat(TPR)-containing SMYD2 and SMYD3.Moreover,using proteomic and biochemical techniques,we provide evidence that SMYD2 methylates K209 and K615 on HSP90 nucleotide-binding and dimerization domains,respectively.In addition,we found that each methylation site displays unique reactivity in regard to the presence of HSP90 co-chaperones,pH,and demethylation by the lysine amine oxidase LSD1,suggesting that alternative mechanisms control HSP90 methylation by SMYD2.Altogether,this study highlights the ability of SMYD proteins to form unique protein complexes that may underlie their various biological functions and the SMYD2-mediated methylation of the key molecular chaperone HSP90.