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      A grape ( Vitis vinifera L.) pomace water extract modulates inflammatory and immune response in SW‐480 cells and isolated mouse colon

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          Abstract

          Grape ( Vitis vinifera L.) pomace is a residue derived from the winemaking process, which contains bioactive compounds displaying noteworthy health‐promoting properties. The aim of the present study was to investigate the phenolic composition and protective effects of a water extract of grape pomace (WEGP) in colorectal cancer cell line SW480 and in isolated mouse colon exposed to Escherichia coli lipopolysaccharide (LPS). The extract decreased SW‐480 cell viability, as well as vascular endothelial factor A (VEGFA), hypoxia‐induced factor 1α (HIF1α), and transient receptor potential M8 (TRPM8) LPS‐induced gene expression. Moreover, the extract inhibited mRNA levels of nuclear factor kB (NFkB), cyclooxygenase (COX)‐2, tumor necrosis factor (TNF)α, interleukin (IL)‐6, IL‐1β, IL‐10, inducible nitric oxide synthase (iNOS), and interferon (IFN)γ, in isolated colon. Conversely, WEGP increased the gene expression of antioxidant catalase (CAT) and superoxide dismutase (SOD), in the same model. The modulatory effects exerted by WEGP could be related, at least in part, to the phenolic composition, with particular regards to the catechin level. Docking calculations also predicted the interactions of catechin toward TRPM8 receptor, deeply involved in colon cancer; thus further suggesting the grape pomace as a valuable source of bioactive extracts and phytochemicals with protective effects in the colon.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            How to calculate sample size in animal studies?

            Calculation of sample size is one of the important component of design of any research including animal studies. If a researcher select less number of animals it may lead to missing of any significant difference even if it exist in population and if more number of animals selected then it may lead to unnecessary wastage of resources and may lead to ethical issues. In this article, on the basis of review of literature done by us we suggested few methods of sample size calculations for animal studies.
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              NF-kappaB in inflammatory bowel disease.

              Apart from genetic and environmental factors, the mucosal immune system of the gut plays a central role in the pathogenesis of inflammatory bowel disease (IBD). In the healthy gut, the mucosal immune system ensures the balance between pro- and anti-inflammatory mediators and thereby allows an effective defence against luminal pathogens but at the same time prevents an overwhelming immune reaction directed against the huge amount of harmless luminal antigens (for example, components of food or nonpathological bacteria). In both entities of IBD (Crohn's disease and ulcerative colitis) this immunological balance is severely impaired and shifted towards the pro-inflammatory side. The chronic mucosal inflammation in IBD is caused by hyperactivation of effector immune cells, which produce high levels of pro-inflammatory cytokines like tumour necrosis factor-alpha, interleukin-6 and interferon-gamma, resulting in colonic tissue damage. The nuclear transcription factor kappaB (NF-kappaB) was identified as one of the key regulators in this immunological setting. Its activation is markedly induced in IBD patients and through its ability to promote the expression of various pro-inflammatory genes, NF-kappaB strongly influences the course of mucosal inflammation. Considering the different cell-type specific effects which are mediated by NF-kappaB, this review aims at describing the complex role of NF-kappaB in IBD and discusses existing pharmacological attempts to block the activation of NF-kappaB to develop new therapeutic strategies in IBD.
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                Author and article information

                Contributors
                giustino.orlando@unich.it
                Journal
                Phytother Res
                Phytother Res
                10.1002/(ISSN)1099-1573
                PTR
                Phytotherapy Research
                John Wiley & Sons, Ltd. (Chichester, UK )
                0951-418X
                1099-1573
                07 September 2022
                December 2022
                : 36
                : 12 ( doiID: 10.1002/ptr.v36.12 )
                : 4620-4630
                Affiliations
                [ 1 ] Department of Pharmacy G. d'Annunzio University of Chieti‐Pescara Chieti Italy
                [ 2 ] Veridia Italia Srl Pescara Italy
                [ 3 ] Department of Pharmacy, School of Medicine University of Naples Federico II Naples Italy
                [ 4 ] Physiology and Biochemistry Laboratory, Department of Biology, Science Faculty Selcuk University Konya Turkey
                Author notes
                [*] [* ] Correspondence

                Giustino Orlando, Department of Pharmacy, G. d'Annunzio University of Chieti‐Pescara, Chieti 66013, Italy.

                Email: giustino.orlando@ 123456unich.it

                Author information
                https://orcid.org/0000-0003-2872-1734
                https://orcid.org/0000-0001-6548-7823
                https://orcid.org/0000-0002-7223-7379
                Article
                PTR7581
                10.1002/ptr.7581
                10087928
                36069605
                b44863ea-f7e8-401d-ae22-1437811fb781
                © 2022 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 07 June 2022
                : 29 March 2022
                : 07 July 2022
                Page count
                Figures: 7, Tables: 3, Pages: 11, Words: 7349
                Funding
                Funded by: National Grant Progetti di Rilevante Interesse Nazionale (PRIN) 2017 from MIUR
                Award ID: 2017XC73BW
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                December 2022
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.2.7 mode:remove_FC converted:11.04.2023

                Pharmacology & Pharmaceutical medicine
                catechin,colon cancer,grape pomace,inflammation,trpm8,vitis vinifera

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