study-of-the-dna-damage-checkpoint-using-xenopus-egg-extracts

Study of the DNA Damage Checkpoint using <em>Xenopus</em> Egg Extracts

On a daily basis, cells are subjected to a variety of endogenous and environmental insults. To combat these insults, cells have evolved DNA damage ...

clinicopathological-analysis-mirna-expression-breast-cancer-tissues

Clinicopathological Analysis of miRNA Expression in Breast Cancer Tissues by Using miRNA <em>In Situ</em> Hybridization

In this article, we describe a detailed protocol for miRNA detection in breast cancer tissue using in situ hybridization with a digoxigenin-labelled LNA ...

University of North Carolina at Charlotte

University of South Carolina

&#x6211;&#x3005; &#x306F;&#x975E; st &#x4E0A;&#x6607;&#x6025;&#x6027;&#x51A0;&#x75C7;&#x5019;&#x7FA4;&#x60A3;&#x8005;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x305F;&#x3081;&#x306E;&#x5B9F;&#x8DF5;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x306B;&#x6551;&#x6025;&#x90E8; (ED) &#x306E;&#x69CB;&#x9020;&#x3068;&#x30B1;&#x30A2;&#x306E;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#x5F71;&#x97FF;&#x3092;&#x8A55;&#x4FA1;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x6C42;&#x3081;&#x305F;&#x3002;

&#x6551;&#x6025;&#x90E8;&#x69CB;&#x9020;&#x3068;&#x30B1;&#x30A2;&#x306E;&#x5F71;&#x97FF;&#x306F;&#x3001;&#x975E; st &#x4E0A;&#x6607;&#x6025;&#x6027;&#x51A0;&#x75C7;&#x5019;&#x7FA4;&#x306E;&#x305F;&#x3081;&#x306E;&#x30B1;&#x30A2;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x4E0A;&#x3067;&#x51E6;&#x7406;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC6B0;&#xB9AC; &#xBE44; ST &#xC138;&#xADF8;&#xBA3C;&#xD2B8; &#xC0C1;&#xC2B9; &#xAE09;&#xC131; &#xAD00;&#xC0C1;&#xB3D9;&#xB9E5; &#xC99D;&#xD6C4;&#xAD70;&#xC73C;&#xB85C; &#xD658;&#xC790; &#xCE58;&#xB8CC;&#xB97C; &#xC704;&#xD55C; &#xC2E4;&#xCC9C; &#xC9C0;&#xCE68; &#xC900;&#xC218;&#xC5D0; &#xBE44;&#xC0C1; &#xBD80;&#xC11C; (ED) &#xAD6C;&#xC870; &#xBC0F; &#xAD00;&#xB9AC; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC758; &#xC601;&#xD5A5;&#xC744; &#xD3C9;&#xAC00; &#xD558;&#xACE0;&#xC790; &#xD588;&#xB2E4;.

&#xC751;&#xAE09;&#xC2E4; &#xAD6C;&#xC870;&#xC640; &#xAD00;&#xB9AC;&#xC758; &#xC601;&#xD5A5; ST &#xC138;&#xADF8;&#xBA3C;&#xD2B8; &#xC0C1;&#xC2B9; &#xAE09;&#xC131; &#xAD00;&#xC0C1;&#xB3D9;&#xB9E5; &#xC99D;&#xD6C4;&#xAD70;&#xC5D0; &#xB300; &#xD55C; &#xCF00;&#xC5B4;&#xB97C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xCC98;&#xB9AC; &#xD569;&#xB2C8;&#xB2E4;.

Se busc&oacute; evaluar la influencia de los procesos de estructura y atenci&oacute;n de urgencias (ED) en adherencia a gu&iacute;as de pr&aacute;ctica para el tratamiento de pacientes con s&iacute;ndrome coronario agudo sin elevaci&oacute;n del segmento ST no.

El impacto de la estructura del servicio de urgencias y cuidado los procesos en la prestaci&oacute;n de cuidado para el s&iacute;ndrome coronario agudo sin elevaci&oacute;n del segmento ST no.

Nous avons cherch&eacute; &agrave; &eacute;valuer l&#39;influence des processus structure et soins d&#39;urgences (ED) sur l&#39;adh&eacute;sion aux lignes directrices de pratique pour le traitement des patients avec un syndrome coronarien aigu &eacute;l&eacute;vation du segment ST non.

L&#39;impact de la structure de service d&#39;urgence et de soins des processus dans la prestation de soins pour les syndromes coronariens aigus &eacute;l&eacute;vation du segment ST non.

Wir versucht, den Einfluss der Notaufnahme (ED)-Aufbau und Pflege-Prozesse zur Einhaltung der Leitlinien f&uuml;r die Behandlung von Patienten mit nicht-ST-Segment Elevation akuten koronaren Syndromen zu bewerten.

Die Auswirkungen der Notaufnahme Struktur und Sorgfalt verarbeitet Lieferung Pflege f&uuml;r nicht-ST-Segment Elevation akuter koronarer syndrome.

Abbiamo cercato di valutare l&#39;influenza dei processi di struttura e cura di reparto di emergenza (ED) in aderenza alle linee guida per il trattamento di pazienti con sindromi coronariche acute non-ST-segmento di elevazione.

L&#39;impatto della struttura del dipartimento d&#39;emergenza e cura i processi nella fornitura di cure per sindromi coronariche acute non-ST-segmento di elevazione.

&#x6211;&#x4EEC;&#x8BBE;&#x6CD5;&#x8BC4;&#x4F30;&#x5BF9;&#x75C5;&#x4EBA;&#x7684;&#x6CBB;&#x7597;&#x5B9E;&#x8DF5;&#x51C6;&#x5219;&#x9075;&#x5B88;&#x975E; ST &#x6BB5;&#x62AC;&#x9AD8;&#x6025;&#x6027;&#x51A0;&#x8109;&#x7EFC;&#x5408;&#x5F81;&#x6025;&#x8BCA;&#x79D1; (ED) &#x7ED3;&#x6784;&#x548C;&#x62A4;&#x7406;&#x8FC7;&#x7A0B;&#x7684;&#x5F71;&#x54CD;&#x3002;

&#x6025;&#x8BCA;&#x79D1;&#x7ED3;&#x6784;&#x548C;&#x62A4;&#x7406;&#x7684;&#x5F71;&#x54CD;&#x8FC7;&#x7A0B;&#x4E2D;&#x63D0;&#x4F9B;&#x5BF9;&#x975E; ST &#x6BB5;&#x62AC;&#x9AD8;&#x6025;&#x6027;&#x51A0;&#x8109;&#x7EFC;&#x5408;&#x5F81;&#x60A3;&#x8005;&#x7684;&#x62A4;&#x7406;&#x3002;

We sought to assess the influence of emergency department (ED) structure and care processes on adherence to practice guidelines for the treatment of patients with non-ST-segment elevation acute coronary syndromes.

The impact of emergency department structure and care processes in delivering care for non-ST-segment elevation acute coronary syndromes.

&#x5728;&#x7CD6;&#x5C3F;&#x75C5;&#x3001; &#x89C6;&#x7F51;&#x819C;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x88AB;&#x6FC0;&#x6D3B;&#x91CA;&#x653E;&#x542F;&#x52A8;&#x795E;&#x7ECF;&#x5143;&#x4E22;&#x5931;&#x548C;&#x7CD6;&#x5C3F;&#x75C5;&#x89C6;&#x7F51;&#x819C;&#x75C5;&#x53D8; (DR) &#x770B;&#x5230;&#x8840;-&#x89C6;&#x7F51;&#x819C;&#x5C4F;&#x969C;&#x7834;&#x574F;&#x7684;&#x708E;&#x6027;&#x7EC6;&#x80DE;&#x56E0;&#x5B50;&#x3002;&#x8BE5;&#x673A;&#x5236;&#x7531;&#x54EA;&#x4E9B;&#x7CD6;&#x5C3F;&#x75C5;&#x6FC0;&#x6D3B;&#x5C0F;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x91CA;&#x653E;&#x8FD9;&#x4E9B;&#x708E;&#x75C7;&#x4ECB;&#x8D28;&#x662F;&#x4E0D;&#x6E05;&#x695A;&#x548C;&#x56E0;&#x6B64;&#x88AB;&#x9610;&#x660E;&#x3002;

&#x89C6;&#x7F51;&#x819C;&#x5C0F;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x6FC0;&#x6D3B;&#x5E76;&#x8BF1;&#x5BFC; amadori &#x7CD6;&#x5316;&#x767D;&#x86CB;&#x767D;&#x5BF9;&#x7CD6;&#x5C3F;&#x75C5;&#x6A21;&#x578B;&#x5927;&#x9F20;&#x7684;&#x708E;&#x75C7;&#x3002;

During diabetes, retinal microglial cells are activated to release inflammatory cytokines that initiate neuronal loss and blood-retinal barrier breakdown seen in diabetic retinopathy (DR). The mechanism by which diabetes activates microglia to release those inflammatory mediators is unclear and was therefore elucidated.

Retinal microglial activation and inflammation induced by amadori-glycated albumin in a rat model of diabetes.

Au cours du diab&egrave;te, les cellules microgliales r&eacute;tiniens sont activ&eacute;s pour lib&eacute;rer des cytokines inflammatoires qui initie la perte neuronale et la barri&egrave;re sang-r&eacute;tinienne ventilation vu dans la r&eacute;tinopathie diab&eacute;tique (RD). Le m&eacute;canisme par lesquels le diab&egrave;te active les cellules microgliales pour lib&eacute;rer ces m&eacute;diateurs de l&#39;inflammation n&#39;est pas clair et a &eacute;t&eacute; &eacute;lucid&eacute;e par cons&eacute;quent.

Activation microgliale r&eacute;tinienne et l&#39;inflammation induite par l&#39;albumine glyqu&eacute;e amadori dans un mod&egrave;le de rat de diab&egrave;te.

Bei Diabetes werden retinale Microglial Zellen aktiviert, um entz&uuml;ndliche Cytokines freizugeben, die Verlust von Nervenzellen und Blut-Retina Schranke Aufschl&uuml;sselung gesehen in Diabetische Retinopathie (DR) zu initiieren. Der Mechanismus von denen Diabetes Mikroglia um diese entz&uuml;ndlichen Mediatoren Freigabe aktiviert ist unklar und wurde daher aufgekl&auml;rt.

Retinale Microglial Aktivierung und Entz&uuml;ndung induziert durch Amadori-glykosyliertem Albumin in einem Rattenmodell f&uuml;r Diabetes.

Durante il diabete, le cellule microgliali della retina sono attivate per rilasciare citochine infiammatorie che avviano la perdita neuronale e la barriera emato-retinica ripartizione visto nella retinopatia diabetica (DR). Il meccanismo da cui diabete attiva microglia per liberare quei mediatori infiammatori &egrave; chiaro e quindi &egrave; stata chiarita.

Attivazione della microglia retinica e infiammazione indotta dall&#39;albumina glicata amadori in un modello del ratto del diabete.

&#x7CD6;&#x5C3F;&#x75C5;&#x4E2D;&#x306B;&#x7DB2;&#x819C;&#x30DF;&#x30AF;&#x30ED;&#x30B0;&#x30EA;&#x30A2;&#x7D30;&#x80DE;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x640D;&#x5931;&#x3068;&#x7CD6;&#x5C3F;&#x75C5;&#x7DB2;&#x819C;&#x75C7; (DR) &#x306B;&#x898B;&#x3089;&#x308C;&#x308B;&#x8840;&#x6DB2;&#x7DB2;&#x819C;&#x30D0;&#x30EA;&#x30A2;&#x5185;&#x8A33;&#x3092;&#x958B;&#x59CB;&#x3059;&#x308B;&#x708E;&#x75C7;&#x6027;&#x30B5;&#x30A4;&#x30C8;&#x30AB;&#x30A4;&#x30F3;&#x3092;&#x653E;&#x51FA;&#x3059;&#x308B;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x306B;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3053;&#x308C;&#x3089;&#x306E;&#x708E;&#x75C7;&#x6027;&#x30E1;&#x30C7;&#x30A3;&#x30A8;&#x30FC;&#x30BF;&#x30FC;&#x3092;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3059;&#x308B;&#x30DF;&#x30AF;&#x30ED;&#x30B0;&#x30EA;&#x30A2;&#x306E;&#x3069;&#x306E;&#x7CD6;&#x5C3F;&#x75C5;&#x3092;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x306B;&#x660E;&#x78BA;&#x3067;&#x3042;&#x308A;&#x3001;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x89E3;&#x660E;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;

&#x7DB2;&#x819C;&#x30DF;&#x30AF;&#x30ED;&#x30B0;&#x30EA;&#x30A2;&#x306E;&#x6D3B;&#x6027;&#x5316;&#x3068;&#x7CD6;&#x5C3F;&#x75C5;&#x306E;&#x30E9;&#x30C3;&#x30C8; &#x30E2;&#x30C7;&#x30EB; amadori &#x7CD6;&#x5316;&#x30A2;&#x30EB;&#x30D6;&#x30DF;&#x30F3;&#x306B;&#x3088;&#x308B;&#x8A98;&#x767A;&#x708E;&#x75C7;&#x3002;

&#xB2F9;&#xB1E8;&#xBCD1;, &#xB3D9;&#xC548; &#xC2E0;&#xACBD; &#xC190;&#xC2E4;&#xACFC; &#xD608;&#xC561; &#xB9DD;&#xB9C9; &#xBC29; &#xBCBD; &#xC1E0; &#xC57D; &#xB2F9;&#xB1E8;&#xBCD1; &#xC131; &#xB9DD;&#xB9C9; &#xC99D; (DR)&#xC5D0;&#xC11C; &#xC2DC;&#xC791; &#xD558;&#xB294; &#xC120; &#xB3D9;&#xC801;&#xC778; cytokines&#xB97C; &#xD480;&#xC5B4; &#xB9DD;&#xB9C9; microglial &#xC138;&#xD3EC; &#xD65C;&#xC131;&#xD654; &#xB429;&#xB2C8;&#xB2E4;. &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC740; &#xC5B4;&#xB5A4; &#xB2F9;&#xB1E8;&#xBCD1; microglia &#xADF8; &#xC5FC;&#xC99D; &#xC131; &#xC911;&#xC7AC;&#xC790; &#xB9B4;&#xB9AC;&#xC2A4;&#xB97C; &#xD65C;&#xC131;&#xD654; &#xD558; &#xC5EC; &#xBA85;&#xD655; &#xD558;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4; &#xD558; &#xACE0; &#xB530;&#xB77C;&#xC11C; &#xD574;&#xBA85; &#xD588;&#xB2E4;.

&#xB9DD;&#xB9C9; microglial &#xD65C;&#xC131;&#xD654; &#xBC0F; amadori-&#xB2F9;&#xD654; &#xD608; &#xB2F9;&#xB1E8;&#xBCD1; &#xC950; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; &#xC54C; &#xBD80; &#xBBFC;&#xC5D0; &#xC758;&#xD574; &#xC720;&#xB3C4; &#xB41C; &#xC5FC;&#xC99D;.

En la diabetes, las c&eacute;lulas microgliales retiniana se activan para liberar citoquinas inflamatorias que inician la p&eacute;rdida neuronal y la ruptura de la Barrera Hematorretinal visto en retinopat&iacute;a diab&eacute;tica (RD). El mecanismo por que diabetes activa microglia para liberar los mediadores inflamatorios es confusa y por lo tanto fue aclarada.

La activaci&oacute;n microglial retiniana y la inflamaci&oacute;n inducida por amadori-glucosilada alb&uacute;mina en un modelo de rata de la diabetes.

The base excision repair pathway is largely responsible for the repair of oxidative stress-induced DNA damage. However, it remains unclear how the DNA damage checkpoint is activated by oxidative stress at the molecular level. Here, we provide evidence showing that hydrogen peroxide (H2O2) triggers checkpoint kinase 1 (Chk1) phosphorylation in an ATR [ataxia-telangiectasia mutated (ATM) and Rad3-related]-dependent but ATM-independent manner in Xenopus egg extracts. A base excision repair protein, Apurinic/apyrimidinic (AP) endonuclease 2 (APE2, APN2, or APEX2), is required for the generation of replication protein A (RPA)-bound single-stranded DNA, the recruitment of a checkpoint protein complex [ATR, ATR-interacting protein (ATRIP), and Rad9] to damage sites, and H2O2-induced Chk1 phosphorylation. A conserved proliferating cell nuclear antigen interaction protein box of APE2 is important for the recruitment of APE2 to H2O2-damaged chromatin. APE2 3'-phosphodiesterase and 3'-5' exonuclease activity is essential for single-stranded DNA generation in the 3'-5' direction from single-stranded breaks, referred to as single-stranded break end resection. In addition, APE2 associates with Chk1, and a serine residue (S86) in the Chk1-binding motif of APE2 is essential for Chk1 phosphorylation, indicating a Claspin-like but distinct role for APE2 in ATR-Chk1 signaling. Our data indicate that APE2 plays a vital and previously unexpected role in ATR-Chk1 checkpoint signaling in response to oxidative stress. Thus, our findings shed light on a distinct mechanism of how an ATR-Chk1-dependent DNA damage checkpoint is mediated by APE2 in the oxidative stress response.

APE2 is required for ATR-Chk1 checkpoint activation in response to oxidative stress.

A previously healthy 22-year-old African American woman presented with bilateral vision loss associated with headache. Her ocular examination was significant for bilateral retinal arterial "boxcarring," retinal whitening, retinal hemorrhages, and cherry red spots. She was diagnosed with bilateral central retinal artery occlusions and was hospitalized due to concomitant diagnosis of stroke and hypercoagulable state. She was also found to be in heart failure and kidney failure. Rheumatology was consulted and she was diagnosed with catastrophic antiphospholipid syndrome in association with systemic lupus erythematosus. Approximately 7 months after presentation, the patient's vision improved and remained stable at 20/200 and 20/80.

Catastrophic antiphospholipid syndrome presenting as bilateral central retinal artery occlusions.

Traumatic head injuries not involving the eye have been known to cause retinal injury through multiple mechanisms. Abusive head trauma remains the prototypical example. We propose to demonstrate the first case of bilateral multiple retinal hemorrhages in a young healthy adult related to riding multiple theme park roller coasters.

ROLLER COASTER RETINOPATHY: CASE REPORT OF SYMPTOMATIC BILATERAL INTRARETINAL HEMORRHAGES AFTER SHAKING INJURY IN AN OTHERWISE HEALTHY ADULT.

Human epidermal growth factor receptor 2 (HER2) is overexpressed/amplified in ∼30% breast cancers which are associated with poor prognosis. microRNAs are small non-coding RNA which play an important role in many physiological conditions including cancer. Here we screened and identified many miRNAs which are dysregulated by HER2 overexpression. In line with our quantitative PCR analysis data, in silico analysis of microRNA expression profiles of 1302 breast tumors revealed that miR-146a-5p is up-regulated and miR-181d and miR-195-5p are down-regulated in HER2-positive tumors. Furthermore, the expression levels of these microRNAs can significantly predict patient survival and thus potentially serve as new prognostic markers for HER2-positive breast cancer.

Identification of new miRNA biomarkers associated with HER2-positive breast cancers.

Human epidermal growth factor receptor 2 (HER2 or ErBb2) is a receptor tyrosine kinase overexpressed in 20-30% of breast cancers and associated with poor prognosis and outcome. Dysregulation of several microRNAs (miRNAs) plays a key role in breast cancer progression and metastasis. In this study, we screened and identified miRNAs dysregualted in HER2-positive breast cancer cells. Our molecular study demonstrated that miR-489 was specifically downregulated by the HER2-downstream signaling, especially through the MAPK pathway. Restoration or overexpression of miR-489 in HER2-positive breast cancer cells significantly inhibited cell growth in vitro and decreased the tumorigenecity and tumor growth in xenograft mice. Mechanistically, we found that overexpression of miR-489 led to the decreased levels of HER2 and SHP2 and thus attenuated HER2-downstream signaling. Furthermore, we for the first time demonstrated that HER2 is a direct target of miR-489 and therefore HER2-SHP2-MAPK and miR-489 signaling pathways form a mutually inhibitory loop. Using quantitative real-time PCR analysis and Fluorescent in situ hybridization technique (FISH), we found that miR-489 was expressed at significantly lower level in tumor tissues compared to the adjacent normal tissues. Downregulation of miR-489 in breast cancers was associated with aggressive tumor phenotypes. Overall, our results define a double-negative feedback loop involving miR-489 and the HER2-SHP2-MAPK signaling axis that can regulate breast cancer cell proliferation and tumor progression and might have therapeutic relevance for HER2-positive breast cancer.

A novel double-negative feedback loop between miR-489 and the HER2-SHP2-MAPK signaling axis regulates breast cancer cell proliferation and tumor growth.

ArticleTotal : 2	Year
Publication title Cited by 35	2012
Publication title Cited by 4	2016

Article	Year
The impact of emergency department structure and care processes in delivering care for non-ST-segment elevation acute coronary syndromes. American heart journal\| PubMed ID: 16996829	2006
Retinal microglial activation and inflammation induced by amadori-glycated albumin in a rat model of diabetes. Diabetes\| PubMed ID: 21317295	2011
APE2 is required for ATR-Chk1 checkpoint activation in response to oxidative stress. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 23754435	2013
Catastrophic antiphospholipid syndrome presenting as bilateral central retinal artery occlusions. Case reports in ophthalmological medicine\| PubMed ID: 25722904	2015
ROLLER COASTER RETINOPATHY: CASE REPORT OF SYMPTOMATIC BILATERAL INTRARETINAL HEMORRHAGES AFTER SHAKING INJURY IN AN OTHERWISE HEALTHY ADULT. Retinal cases & brief reports\| PubMed ID: 26630245	2016
Identification of new miRNA biomarkers associated with HER2-positive breast cancers. Oncoscience\| PubMed ID: 26697527	2015
A novel double-negative feedback loop between miR-489 and the HER2-SHP2-MAPK signaling axis regulates breast cancer cell proliferation and tumor growth. Oncotarget\| PubMed ID: 26918448	2016

Yogin Patel

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