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https://www.readbyqxmd.com/read/27910942/improved-bi-allelic-modification-of-a-transcriptionally-silent-locus-in-patient-derived-ipsc-by-cas9-nickase
#1
Reto Eggenschwiler, Mohsen Moslem, Mariane Serra Fráguas, Melanie Galla, Oliver Papp, Maximilian Naujock, Ines Fonfara, Ingrid Gensch, Annabell Wähner, Abbas Beh-Pajooh, Claudio Mussolino, Marcel Tauscher, Doris Steinemann, Florian Wegner, Susanne Petri, Axel Schambach, Emmanuelle Charpentier, Toni Cathomen, Tobias Cantz
Homology directed repair (HDR)-based genome editing via selectable long flanking arm donors can be hampered by local transgene silencing at transcriptionally silent loci. Here, we report efficient bi-allelic modification of a silent locus in patient-derived hiPSC by using Cas9 nickase and a silencing-resistant donor construct that contains an excisable selection/counter-selection cassette. To identify the most active single guide RNA (sgRNA)/nickase combinations, we employed a lentiviral vector-based reporter assay to determine the HDR efficiencies in cella...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27905063/efficient-gene-targeting-in-mouse-zygotes-mediated-by-crispr-cas9-protein
#2
Chris J Jung, Junli Zhang, Elizabeth Trenchard, Kent C Lloyd, David B West, Barry Rosen, Pieter J de Jong
The CRISPR/Cas9 system has rapidly advanced targeted genome editing technologies. However, its efficiency in targeting with constructs in mouse zygotes via homology directed repair (HDR) remains low. Here, we systematically explored optimal parameters for targeting constructs in mouse zygotes via HDR using mouse embryonic stem cells as a model system. We characterized several parameters, including single guide RNA cleavage activity and the length and symmetry of homology arms in the construct, and we compared the targeting efficiency between Cas9, Cas9nickase, and dCas9-FokI...
November 30, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27899664/integration-defective-lentiviral-vector-mediates-efficient-gene-editing-through-homology-directed-repair-in-human-embryonic-stem-cells
#3
Yebo Wang, Yingjia Wang, Tammy Chang, He Huang, Jiing-Kuan Yee
Human embryonic stem cells (hESCs) are used as platforms for disease study, drug screening and cell-based therapy. To facilitate these applications, it is frequently necessary to genetically manipulate the hESC genome. Gene editing with engineered nucleases enables site-specific genetic modification of the human genome through homology-directed repair (HDR). However, the frequency of HDR remains low in hESCs. We combined efficient expression of engineered nucleases and integration-defective lentiviral vector (IDLV) transduction for donor template delivery to mediate HDR in hESC line WA09...
November 28, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27899012/disease-outcomes-and-care-fragmentation-among-patients-with-systemic-lupus-erythematosus
#4
Theresa L Walunas, Kathryn L Jackson, Anh H Chung, Karen A Mancera-Cuevas, Daniel L Erickson, Rosalind Ramsey-Goldman, Abel Kho
OBJECTIVE: To examine the impact of care fragmentation across multiple healthcare institutions on disease outcomes in patients with systemic lupus erythematosus (SLE). METHODS: Using the Chicago HealthLNK Data Repository (HDR), an assembly of electronic health records from six institutions, we identified patients with SLE, using ICD-9 codes, whose care was delivered at more than one organization. We examined whether patients had severe infections or comorbidities (ICD-9 code defined) that indicate SLE-induced damage...
November 29, 2016: Arthritis Care & Research
https://www.readbyqxmd.com/read/27898095/efficient-generation-of-orthologous-point-mutations-in-pigs-via-crispr-assisted-ssodn-mediated-homology-directed-repair
#5
Kankan Wang, Xiaochun Tang, Yan Liu, Zicong Xie, Xiaodong Zou, Mengjing Li, Hongming Yuan, Hongsheng Ouyang, Huping Jiao, Daxin Pang
Precise genome editing in livestock is of great value for the fundamental investigation of disease modeling. However, genetically modified pigs carrying subtle point mutations were still seldom reported despite the rapid development of programmable endonucleases. Here, we attempt to investigate single-stranded oligonucleotides (ssODN) mediated knockin by introducing two orthologous pathogenic mutations, p.E693G for Alzheimer's disease and p.G2019S for Parkinson's disease, into porcine APP and LRRK2 loci, respectively...
November 29, 2016: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/27895687/commissioning-and-quality-assurance-procedures-for-the-hdr-valencia-skin-applicators
#6
REVIEW
Domingo Granero, Cristian Candela-Juan, Facundo Ballester, Zoubir Ouhib, Javier Vijande, Jose Richart, Jose Perez-Calatayud
The Valencia applicators (Nucletron, an Elekta company, Elekta AB, Stockholm, Sweden) are cup-shaped tungsten applicators with a flattening filter used to collimate the radiation produced by a high-dose-rate (HDR) (192)Ir source, and provide a homogeneous absorbed dose at a given depth. This beam quality provides a good option for the treatment of skin lesions at shallow depth (3-4 mm). The user must perform commissioning and periodic testing of these applicators to guarantee the proper and safe delivery of the intended absorbed dose, as recommended in the standards in radiation oncology...
October 2016: Journal of Contemporary Brachytherapy
https://www.readbyqxmd.com/read/27895685/pediatric-peri-operative-fractionated-high-dose-rate-brachytherapy-for-recurrent-wilms-tumor-using-a%C3%A2-reconstructed-freiburg-flap
#7
Emily Flower, Kathy Ngoc Tran, Salman Zanjani, Wayne Smith, Jonathan Karpelowsky, Katie Summerhayes, Edgar Estoesta, Jennifer Chard
PURPOSE: To report peri-operative fractionated high-dose-rate (HDR) brachytherapy with a 3D customized Freiburg flap applicator to treat locally recurrent Wilms' tumor, followed by immediate hyperthermic intraperitoneal chemotherapy for a 16-year-old with a second recurrence of nephroblastoma (Wilms' tumor). MATERIAL AND METHODS: The tumor was excised and surgical bed was treated with fractionated HDR brachytherapy using a Freiburg flap applicator. Hyperthermic intraperitoneal chemotherapy was performed immediately after the removal of brachytherapy applicator...
October 2016: Journal of Contemporary Brachytherapy
https://www.readbyqxmd.com/read/27895683/salvage-interstitial-brachytherapy-based-on-computed-tomography-for-recurrent-cervical-cancer-after-radical-hysterectomy-and-adjuvant-radiation-therapy-case-presentations-and-introduction-of-the-technique
#8
Zhong-Shan Liu, Jie Guo, Yang-Zhi Zhao, Xia Lin, Bin Chen, Ming Zhang, Jiang-Ming Li, Xiao-Jun Ren, Bing-Ya Zhang, Tie-Jun Wang
PURPOSE: Locally recurring cervical cancer after surgery and adjuvant radiotherapy remains a major therapeutic challenge. This paper presents a new therapeutic technique for such patients: interstitial brachytherapy (BT) guided by real-time three-dimensional (3D) computed tomography (CT). MATERIAL AND METHODS: Sixteen patients with recurrent cervical cancer after radical surgery and adjuvant external-beam radiotherapy (EBRT) were included in this study. These patients underwent high-dose-rate (HDR) interstitial BT with free-hand placement of metal needles guided by real-time 3D-CT...
October 2016: Journal of Contemporary Brachytherapy
https://www.readbyqxmd.com/read/27895680/the-role-of-interstitial-brachytherapy-in-the-management-of-primary-radiation-therapy-for-uterine-cervical-cancer
#9
Naoya Murakami, Kazuma Kobayashi, Tomoyasu Kato, Satoshi Nakamura, Akihisa Wakita, Hiroyuki Okamoto, Satoshi Shima, Keisuke Tsuchida, Tairo Kashihara, Ken Harada, Kana Takahashi, Rei Umezawa, Koji Inaba, Yoshinori Ito, Hiroshi Igaki, Jun Itami
PURPOSE: The aim of this study was to report the clinical results of uterine cervical cancer patients treated by primary radiation therapy including brachytherapy, and investigate the role of interstitial brachytherapy (ISBT). MATERIAL AND METHODS: All consecutive uterine cervical cancer patients who were treated by primary radiation therapy were reviewed, and those who were treated by ISBT were further investigated for clinical outcomes and related toxicities. RESULTS: From December 2008 to October 2014, 209 consecutive uterine cervical cancer patients were treated with primary radiation therapy...
October 2016: Journal of Contemporary Brachytherapy
https://www.readbyqxmd.com/read/27885590/payment-reform-needed-to-address-health-disparities-of-undiagnosed-diabetic-retinopathy-in-the-city-of-chicago
#10
Dustin D French, Jess J Behrens, Kathryn L Jackson, Abel N Kho, Theresa L Walunas, Charlesnika T Evans, Michael Mbagwu, Curtis E Margo, Paul J Bryar
INTRODUCTION: The Affordable Care Act (ACA) has expanded health coverage for thousands of Illinois residents. Expanded coverage, however, does not guarantee appropriate health care. Diabetes and its ocular complications serve as an example of how providers in underserved urban areas may not be able to keep up with new demand for labor- and technology-intensive health care unless changes in reimbursement policies are instituted. METHODS: A retrospective cohort study was conducted using medical encounter information from the Chicago HealthLNK Data Repository (HDR), an assembly of non-duplicated and de-identified patient medical records...
November 24, 2016: Ophthalmology and Therapy
https://www.readbyqxmd.com/read/27882806/comorbidity-as-a-predictor-of-overall-survival-in-prostate-cancer-patients-treated-with-external-beam-radiotherapy-combined-with-hdr-brachytherapy-boosts
#11
Marie Hjälm-Eriksson, Anders Ullén, Hemming Johansson, Seymoure Levitt, Sten Nilsson, Karl-Mikael Kälkner
BACKGROUND: The risk stratification currently applied prior to curative treatment for localized prostate cancer (PC) does not take into account comorbidity or age. Therefore, we investigated the impact of comorbidity on overall survival (OS) in PC patients treated with external beam radiotherapy (EBRT) and high-dose rate (HDR) brachytherapy boost. MATERIAL AND METHODS: At a single center, 611 consecutive patients diagnosed with localized PC from 1998 to 2004 underwent definitive EBRT (50 Gy) and HDR brachytherapy boosts (2 × 10 Gy) combined with neoadjuvant total androgen blockade...
November 24, 2016: Acta Oncologica
https://www.readbyqxmd.com/read/27876410/clinical-outcome-of-high-dose-rate-interstitial-brachytherapy-in-vulvar-cancer-a-single-institutional-experience
#12
Umesh Mahantshetty, Pushpa Naga, Reena Engineer, Supriya Sastri, Yogesh Ghadi, Udita Upreti, Vijaya Somesan, Sudarshan Kadam, Satish Kohle, Deepak Deshpande, Shyam Kishore Shrivastava
PURPOSE: With an aim to evaluate and report high dose date interstitial brachytherapy (HDR-ISBT) in vulvar cancers, we undertook this retrospective analysis. METHODS AND MATERIALS: Histologically proven vulvar cancers treated with HDR-ISBT between 2001 and 2016 were analyzed. Radiotherapy details, clinical outcome in terms of local control rates, survivals, and toxicities were evaluated. RESULTS: A total of 38 patients received HDR-ISBT, with definitive radiation in 29 (76...
November 19, 2016: Brachytherapy
https://www.readbyqxmd.com/read/27869163/wwox-brca1-interaction-role-in-dna-repair-pathway-choice
#13
M S Schrock, B Batar, J Lee, T Druck, B Ferguson, J H Cho, K Akakpo, H Hagrass, N A Heerema, F Xia, J D Parvin, C M Aldaz, K Huebner
In this study, loss of expression of the fragile site-encoded Wwox protein was found to contribute to radiation and cisplatin resistance of cells, responses that could be associated with cancer recurrence and poor outcome. WWOX gene deletions occur in a variety of human cancer types, and reduced Wwox protein expression can be detected early during cancer development. We found that Wwox loss is followed by mild chromosome instability in genomes of mouse embryo fibroblast cells from Wwox-knockout mice. Human and mouse cells deficient for Wwox also exhibit significantly enhanced survival of ionizing radiation and bleomycin treatment, agents that induce double-strand breaks (DSBs)...
November 21, 2016: Oncogene
https://www.readbyqxmd.com/read/27866953/triple-tandem-high-dose-rate-brachytherapy-for-early-stage-medically-inoperable-endometrial-cancer-initial-report-on-acute-toxicity-and-dosimetric-comparison-to-stereotactic-body-radiation-therapy
#14
Greg Kauffmann, Tianming Wu, Hania Al-Hallaq, Yasmin Hasan
PURPOSE: Stereotactic body radiotherapy (SBRT) may be appealing in medically inoperable endometrial cancer to avoid procedural risks. We performed a dosimetric comparison to triple-tandem, high-dose-rate (HDR) brachytherapy. METHODS AND MATERIALS: Six consecutive clinical stage I, grade 1-2, medically inoperable endometrial cancer patients were treated with triple-tandem HDR brachytherapy. We report patient factors and acute toxicity. Also, we performed dosimetric comparison to SBRT using both 3D conformal arc (3DArc) and volumetric-modulated arc therapy...
November 17, 2016: Brachytherapy
https://www.readbyqxmd.com/read/27864278/integrative-analysis-for-the-role-of-long-non-coding-rnas-in-radiation-induced-mouse-thymocytes-responses
#15
Hui Gao, Zhuo Dong, Wei Wei, Lihong Shao, Linlin Jin, Yahui Lv, Gang Zhao, Shunzi Jin
Long non-coding RNAs (lncRNAs) are a critical class of regulatory molecules involved in a variety of biological functions; however, their role in immune cells response to radiation is unknown. Therefore, in this study we used integrative analysis to determine the expression profile of lncRNAs in mouse thymocytes and the potential functions of lncRNAs in response to radiation. Microarray data profiling indicated that 53 lncRNAs (36 up-regulated and 17 down-regulated) and 74 coding genes (39 up-regulated and 35 down-regulated) were highly differentially expressed in the high dose radiation (HDR) group compared with the control group...
November 17, 2016: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/27846172/colon-cancer-surgery-a-retrospective-study-based-on-a-large-administrative-database
#16
Mario Saia, Alessandra Buja, Gino Sartor, Domenico Mantoan, Ferdinando Agresta, Vincenzo Baldo
INTRODUCTION: Laparoscopic colectomy (LC) for colon cancer is considered as safe and effective as open colectomy. It has many benefits in terms of postoperative pain, length of hospital stay, and esthetic results. There is evidence in the United States of a positive trend in the proportion of oncologic patients undergoing LC. Our aim is to confirm this trend in Europe and identify factors associated with a greater use of LC. MATERIALS AND METHODS: A retrospective cohort study based on hospital discharge records (HDR) collected from 2008 to 2013 in a north-eastern Italian region (Veneto)...
November 14, 2016: Surgical Laparoscopy, Endoscopy & Percutaneous Techniques
https://www.readbyqxmd.com/read/27845913/bladder-accumulated-dose-in-image-guided-high-dose-rate-brachytherapy-for-locally-advanced-cervical-cancer-and-its-relation-to-urinary-toxicity
#17
Roja Zakariaee, Ghassan Hamarneh, Colin J Brown, Marc Gaudet, Christina Aquino-Parsons, Ingrid Spadinger
The purpose of this study was to estimate locally accumulated dose to the bladder in multi-fraction high-dose-date (HDR) image-guided intracavitary brachytherapy (IG-ICBT) for cervical cancer, and study the locally-accumulated dose parameters as predictors of late urinary toxicity. A retrospective study of 60 cervical cancer patients who received five HDR IG-ICBT sessions was performed. The bladder outer and inner surfaces were segmented for all sessions and a bladder-wall contour point-set was created in MATLAB...
December 21, 2016: Physics in Medicine and Biology
https://www.readbyqxmd.com/read/27845164/enhanced-crispr-cas9-mediated-precise-genome-editing-by-improved-design-and-delivery-of-grna-cas9-nuclease-and-donor-dna
#18
Xiquan Liang, Jason Potter, Shantanu Kumar, Namritha Ravinder, Jonathan D Chesnut
While CRISPR-based gene knock out in mammalian cells has proven to be very efficient, precise insertion of genetic elements via the cellular homology directed repair (HDR) pathway remains a rate-limiting step to seamless genome editing. Under the conditions described here, we achieved up to 56% targeted integration efficiency with up to a six-nucleotide insertion in HEK293 cells. In induced pluripotent stem cells (iPSCs), we achieved precise genome editing rates of up to 45% by co-delivering the Cas9 RNP and donor DNA...
November 11, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27836667/exploring-the-potential-of-genome-editing-crispr-cas9-technology
#19
REVIEW
Vijai Singh, Darren Braddick, Pawan Kumar Dhar
CRISPR-Cas9 is an RNA-mediated adaptive immune system that protects bacteria and archaea from viruses or plasmids. Herein we discuss the recent development of CRISPR-Cas9 into a key technology for genome editing, targeting, and regulation in a wide range of organisms and cell types. It requires a custom designed single guide-RNA (sgRNA), a Cas9 endonuclease, and PAM sequences in the target region. The sgRNA-Cas9 complex binds to its target and creates a double-strand break (DSB) that can be repaired by non-homologous end joining (NHEJ) or by the homology-directed repair (HDR) pathway, modifying or permanently replacing the genomic target sequence...
November 9, 2016: Gene
https://www.readbyqxmd.com/read/27834929/prostate-specific-antigen-psa-as-predicting-marker-for-clinical-outcome-and-evaluation-of-early-toxicity-rate-after-high-dose-rate-brachytherapy-hdr-bt-in-combination-with-additional-external-beam-radiation-therapy-ebrt-for-high-risk-prostate-cancer
#20
Thorsten H Ecke, Hui-Juan Huang-Tiel, Klaus Golka, Silvia Selinski, Berit Christine Geis, Stephan Koswig, Katrin Bathe, Steffen Hallmann, Holger Gerullis
High-dose-rate brachytherapy (HDR-BT) with external beam radiation therapy (EBRT) is a common treatment option for locally advanced prostate cancer (PCa). Seventy-nine male patients (median age 71 years, range 50 to 79) with high-risk PCa underwent HDR-BT following EBRT between December 2009 and January 2016 with a median follow-up of 21 months. HDR-BT was administered in two treatment sessions (one week interval) with 9 Gy per fraction using a planning system and the Ir192 treatment unit GammaMed Plus iX. EBRT was performed with CT-based 3D-conformal treatment planning with a total dose administration of 50...
November 10, 2016: International Journal of Molecular Sciences
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