Klinisk prövning på Cervix Cancer: Cervical tissue biopsy

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Measurement of H2AX Phosphorylation as a Marker of Ionizing Radiation Induced Cell Damage Aida Muslimovic, Pegah Johansson and Ola Hammarsten * Department of Clinical Chemis try and Transfusion Medicine, The Institute of Biomedicine, The Sahlgren ska Academy at University of Gothenburg, Sweden 1. Introduction the H2AX molecule is phosphorylated to produce γH2AX, which in recent years, has emerged as one of the most well established chromatin modifications linked to DNA damage and repair. 9,10. Phosphorylation occurs at the highly conserved SQ motif of H2AX, a common substrate for the phosphatidyl-inosito 3-kinase Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174) This image is courtesy of an anonymous Abreview.

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It plays an important role in localisation and structuring of the repair focus. H2AX also plays an active role in supplying the active repair proteins. γH2AX participates in DSB repair. This protein serves as a signal and target of phosphorylation in the spreading phase. Medvedeva, N. G., Panyutin, I. V., Panyutin, I. G. and Neumann, R. D. Phosphorylation of Histone H2AX in Radiation-Induced Micronuclei. Radiat. Res. 168, 493–498 (2007).DNA double-strand breaks are thought to precede the formation of most radiation-induced micronuclei.

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Because phosphorylation of H2AX at Ser 139 (γ-H2AX) is abundant, fast, and correlates well with each DSB, it is the most sensitive marker that can be used to examine the DNA damage produced and the subsequent repair of the DNA lesion. γ-H2AX can be detected by immunoblotting and immunostaining using microscopic or flow cytometric detection. Phosphorylation of the Ser-139 residue of the histone variant H2AX, forming γH2AX, is an early cellular response to the induction of DNA double-strand breaks.

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Here we describe a rapid and simple flow-cytometry-based method, optimized to measure gamma-H2AX in non-fixed peripheral blood cells. The phosphorylation of H2AX can be detected by Western blotting or immunofluorescence, revealing the frequency of DSBs. The phosphatidylinositol 3-kinases have been implicated in H2AX phosphorylation, but it is unclear if ATM is the primary H2AX kinase or if other members of the family such as DNA-PK and ATR contribute in a similar manner. H2AX phosphorylation was evaluated as a marker for genotoxicity in two different murine cell types. This study used two different methods of assessing H2AX phosphorylation to assess the response to genotoxic and non-genotoxic agents: microscope-based manual scoring of γH2AX foci and a high-throughput method using flow cytometry.

Activity of 𝛾 H2AX could be summarized to following main points: accumulation of DNA damage signaling and repair proteins at DSBs The generation of DSBs triggers the relocalization of many DNA damage response (DDR) proteins such as MRE11/NBS1/RAD50, MDC1 Phosphorylation of many repair proteins has been shown to be important for their recruitment to DNA damage-induced intra-nuclear foci. Further, phosphorylation of H2AX (gamma-H2AX) has been shown to be important for either the recruitment or stable retention of DNA repair proteins to these intra-nuclear foci. H2AX becomes phosphorylated on serine 139, then called γH2AX, as a reaction on DNA double-strand breaks (DSB).
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Our results show that the DNA damage response pathway, including γ-H2AX and that brimonidine triggered a robust and transient phosphorylation of ERK1/2.

TP53) as well as higher resulting in SYK phosphorylation and recruitment of. additional targets of IFNγand/or IFNγ/STAT1 signaling and upregu-. lation of NF-κB  PP2A as regulator of intracellular protein phosphorylation (inaktivt) 2008-01-​17, H2AX-metoden för mätning av DNA-skador efter exponering för 2007-01-​22, Celldödsmekanismer i Alzheimers sjukdom: Studier av γ-sekretasaktivitet i  alpha, IFN-gamma-inducible protein 10 (IP-10), IL-lb, his tamine], and Gamma-​aminobutyric acid (GABA).
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DNA-replikering - DNA re-replication - qaz.wiki

Gamma H2AX and its phosphorylation can be detected through western blotting and immunofluorescence, serving as a biomarker for DSB frequency (1-4). Ataxia telangiectasia mutated (ATM) is considered a key protein for H2AX phosphorylation (1-4). 2010-03-06 · DNA DSB induced by gamma irradiation, or other DSB inducers, leads to rapid phosphorylation of H2AX at Ser139 by ATM, ATR and DNA-PKcs, resulting in γH2AX . The foci formed by γH2AX can recruit DNA damage response proteins, such as BRCA1, 53BP1, and MDC1, to initiate DNA repair [ 5 , 27 , 28 ]. Histone H2AX rapidly undergoes phosphorylation at Ser139 (gamma-H2AX) in response to DNA double-strand breaks. Although ATM kinase and DNA-PK phosphorylate Ser139 of H2AX in culture cells, the regulatory mechanism of gamma-H2AX level remains unclear in vivo. Here, we detected the phosphorylation of H2AX and the elimination of gamma-H2AX in the mouse skin after X-irradiation.