Digital Commons Network^™

What Clinical Trials Are Needed For Treatment Of Leiomyosarcoma?, Bernd Kasper, Lorenzo D'Ambrosio, Elizabeth J Davis, Matthew Ingham, Javier Martin Broto, Jonathan C Trent, Winan J Van Houdt, Brian A Van Tine

2020-Current year OA Pubs

Leiomyosarcoma is one of the most common subtypes of soft tissue sarcomas accounting for approximately 20% of sarcomas. As leiomyosarcoma patients frequently develop metastatic disease, effective systemic therapies are needed to improve clinical outcomes. The overall activity of the currently available conventional systemic therapies and the prognosis of patients with advanced and/or metastatic disease are poor. As such, the treatment of this patient population remains challenging. As a result, there is a clear unmet medical need, and designing and performing meaningful clinical studies are of utmost importance to improve the prognosis of this patient group. Therefore, the aim of this …

Reproducible Brain-Wide Association Studies Require Thousands Of Individuals, Scott Marek, David F Montez, Benjamin P Kay, Alexander S Hatoum, Meghan Rose Donohue, Ryland L Miller, Sridhar Kandala, Dillan J Newbold, Annie Zheng, Nicole A Seider, Andrew N Van, Athanasia Metoki, Timothy O Laumann, Steven E Petersen, Deanna M Barch, Nico U F Dosenbach, Et Al.

2020-Current year OA Pubs

Magnetic resonance imaging (MRI) has transformed our understanding of the human brain through well-replicated mapping of abilities to specific structures (for example, lesion studies) and functions

Trabecular Meshwork Mitochondrial Function And Oxidative Stress: Clues To Racial Disparities Of Glaucoma, Hongli Wu, Ying-Bo Shui, Ying Liu, Xiaobin Liu, Carla J Siegfried

2020-Current year OA Pubs

PURPOSE: To identify racial differences of oxidative damage and stress and mitochondrial function in human trabecular meshwork (TM).

DESIGN: Experimental study.

PARTICIPANTS: One hundred seventy-three eyes of 173 patients undergoing intraocular surgery provided aqueous humor (AH) for analysis. Trabecular meshwork tissues from eye bank donors were used as healthy controls for primary cell culture.

METHODS: Enzyme-linked immunosorbent assay methods were used to measure 8-hydroxy-2-deoxyguanosine (8-OHdG), an oxidative damage marker, in AH comparing Black and White Americans. Human TM primary cultured cells from Black and White donors were used for adenosine triphosphate (ATP) measurement under high and low oxygen culture conditions. …

Numbers And Narratives: How Qualitative Methods Can Strengthen The Science Of Paediatric Antimicrobial Stewardship, Charlotte Z Woods-Hill, Anping Xie, John Lin, Heather A Wolfe, Alex S Plattner, Sara Malone, Kathleen Chiotos, Julia E Szymczak

2020-Current year OA Pubs

Antimicrobial and diagnostic stewardship initiatives have become increasingly important in paediatric settings. The value of qualitative approaches to conduct stewardship work in paediatric patients is being increasingly recognized. This article seeks to provide an introduction to basic elements of qualitative study designs and provide an overview of how these methods have successfully been applied to both antimicrobial and diagnostic stewardship work in paediatric patients. A multidisciplinary team of experts in paediatric infectious diseases, paediatric critical care and qualitative methods has written a perspective piece introducing readers to qualitative stewardship work in children, intended as an overview to highlight the importance …

Moving The Needle: Directed Intervention By The American Society For Surgery Of The Hand Is Effective In Encouraging Diversity In Expert Panel Composition, Lauren Elisabeth Wessel, Ambika E Paulson, Elizabeth A Graesser, Amy M Moore, Christopher J Dy

2020-Current year OA Pubs

PURPOSE: Recent efforts have been made by the American Society for Surgery of the Hand to encourage female inclusion in expert panels. We hypothesized that female representation on expert panels has increased over the past decade and that a directed intervention by the American Society for Surgery of the Hand would be associated with an increased percentage of submissions with female panelists.

METHODS: We performed a retrospective analysis of Instructional Course Lecture and Symposium submissions for the 2011 through 2021 American Society for Surgery of the Hand Annual Meetings. Authorship was reviewed, and the gender of the proposed authors was …

Dmp1 Lineage Cells Contribute Significantly To Periosteal Lamellar Bone Formation Induced By Mechanical Loading But Are Depleted From The Bone Surface During Rapid Bone Formation, Taylor L Harris, Matthew J Silva

2020-Current year OA Pubs

Previous work has shown that osteoprogenitor cells (Prx1+) and pre-osteoblasts (Osx+) contribute to mechanical loading-induced bone formation. However, the role of mature Dmp1-expressing osteoblasts has not been reported. In this study we assessed the contribution of osteoblast lineage cells to bone formation at an early time point following mechanical loading (day 8 from onset of loading). We labeled Osx-expressing and Dmp1-expressing cells in inducible Osx and Dmp1 reporter mice (iOsx-Ai9, iDmp1-Ai9), respectively, 3 weeks before loading. Mice were then loaded daily for 5 days (days 1-5) and were dosed with 5-ethynyl-2'-deoxyuridine (EdU) in their drinking water until euthanasia on day …

Risk Of Intracranial Haemorrhage And Ischaemic Stroke After Convexity Subarachnoid Haemorrhage In Cerebral Amyloid Angiopathy: International Individual Patient Data Pooled Analysis, Isabel Charlotte Hostettler, Diane Aum, Gregory Joseph Zipfel, Et Al.

2020-Current year OA Pubs

OBJECTIVE: To investigate the frequency, time-course and predictors of intracerebral haemorrhage (ICH), recurrent convexity subarachnoid haemorrhage (cSAH), and ischemic stroke after cSAH associated with cerebral amyloid angiopathy (CAA).

METHODS: We performed a systematic review and international individual patient-data pooled analysis in patients with cSAH associated with probable or possible CAA diagnosed on baseline MRI using the modified Boston criteria. We used Cox proportional hazards models with a frailty term to account for between-cohort differences.

RESULTS: We included 190 patients (mean age 74.5 years; 45.3% female) from 13 centers with 385 patient-years of follow-up (median 1.4 years). The risks of each …

Pharmacological Chaperones For The Oxytocin Receptor Increase Oxytocin Responsiveness In Myometrial Cells, Manasi Malik, Yingye Fang, Monali Wakle-Prabagaran, Michelle Roh, Kevin Prifti, Antonina I Frolova, Princess I Imoukhuede, Sarah K England

2020-Current year OA Pubs

Oxytocin is a potent uterotonic agent administered to nearly all patients during childbirth in the United States. Inadequate oxytocin response can necessitate Cesarean delivery or lead to uterine atony and postpartum hemorrhage. Thus, it may be clinically useful to identify patients at risk for poor oxytocin response and develop strategies to sensitize the uterus to oxytocin. Previously, we showed that the V281M variant in the oxytocin receptor (OXTR) gene impairs OXTR trafficking to the cell surface, leading to a decreased oxytocin response in cells. Here, we sought to identify pharmacological chaperones that increased oxytocin response in cells expressing WT or …

Care Team And Practice-Level Implementation Strategies To Optimize Pediatric Collaborative Care: Study Protocol For A Cluster-Randomized Hybrid Type Iii Trial, David J Kolko, Elizabeth A Mcguier, Renee Turchi, Eileen Thompson, Satish Iyengar, Shawna N Smith, Kimberly Hoagwood, Celeste Liebrecht, Ian M Bennett, Byron J Powell, Kelly Kelleher, Maria Silva, Amy M Kilbourne

2020-Current year OA Pubs

BACKGROUND: Implementation facilitation is an effective strategy to support the implementation of evidence-based practices (EBPs), but our understanding of multilevel strategies and the mechanisms of change within the "black box" of implementation facilitation is limited. This implementation trial seeks to disentangle and evaluate the effects of facilitation strategies that separately target the care team and leadership levels on implementation of a collaborative care model in pediatric primary care. Strategies targeting the provider care team (TEAM) should engage team-level mechanisms, and strategies targeting leaders (LEAD) should engage organizational mechanisms.

METHODS: We will conduct a hybrid type 3 effectiveness-implementation trial in a …

Acquired Hydrocephalus Is Associated With Neuroinflammation, Progenitor Loss, And Cellular Changes In The Subventricular Zone And Periventricular White Matter, Maria Garcia-Bonilla, Leandro Castaneyra-Ruiz, Sarah Zwick, Michael Talcott, Ayodamola Otun, Albert M Isaacs, Diego M Morales, David D Limbrick, James P Mcallister

2020-Current year OA Pubs

BACKGROUND: Hydrocephalus is a neurological disease with an incidence of 80-125 per 100,000 births in the United States. Neuropathology comprises ventriculomegaly, periventricular white matter (PVWM) alterations, inflammation, and gliosis. We hypothesized that hydrocephalus in a pig model is associated with subventricular and PVWM cellular alterations and neuroinflammation that could mimic the neuropathology described in hydrocephalic infants.

METHODS: Hydrocephalus was induced by intracisternal kaolin injections in 35-day old female pigs (n = 7 for tissue analysis, n = 10 for CSF analysis). Age-matched sham controls received saline injections (n = 6). After 19-40 days, MRI scanning was performed to measure the …

Artificial Intelligence In Mammographic Phenotyping Of Breast Cancer Risk: A Narrative Review, Aimilia Gastounioti, Shyam Desai, Vinayak S Ahluwalia, Emily F Conant, Despina Kontos

2020-Current year OA Pubs

BACKGROUND: Improved breast cancer risk assessment models are needed to enable personalized screening strategies that achieve better harm-to-benefit ratio based on earlier detection and better breast cancer outcomes than existing screening guidelines. Computational mammographic phenotypes have demonstrated a promising role in breast cancer risk prediction. With the recent exponential growth of computational efficiency, the artificial intelligence (AI) revolution, driven by the introduction of deep learning, has expanded the utility of imaging in predictive models. Consequently, AI-based imaging-derived data has led to some of the most promising tools for precision breast cancer screening.

MAIN BODY: This review aims to synthesize the …

Dietary Patterns Are Associated With Improved Ovarian Reserve In Overweight And Obese Women: A Cross-Sectional Study Of The Lifestyle And Ovarian Reserve (Lore) Cohort, Ashley M Eskew, Bronwyn S Bedrick, Jorge E Chavarro, Joan K Riley, Emily S Jungheim

2020-Current year OA Pubs

BACKGROUND: Growing evidence suggests that adherence to certain dietary patterns is associated with improved fecundity and reproductive outcomes in the general population and infertile couples assisted reproductive treatments. The objective of this study was to assess if dietary patterns are associated with ovarian reserve in reproductive age women without a history of infertility.

METHODS: This was a cross-sectional study of 185 women in the Lifestyle and Ovarian Reserve (LORe) cohort. Women aged 18-44 without a history of infertility were recruited from the local community at an academic medical center. Subjects completed validated food frequency and physical activity questionnaires to assess …

The Social Cerebellum: A Large-Scale Investigation Of Functional And Structural Specificity And Connectivity, Athanasia Metoki, Yin Wang, Ingrid R Olson

2020-Current year OA Pubs

The cerebellum has been traditionally disregarded in relation to nonmotor functions, but recent findings indicate it may be involved in language, affective processing, and social functions. Mentalizing, or Theory of Mind (ToM), is the ability to infer mental states of others and this skill relies on a distributed network of brain regions. Here, we leveraged large-scale multimodal neuroimaging data to elucidate the structural and functional role of the cerebellum in mentalizing. We used functional activations to determine whether the cerebellum has a domain-general or domain-specific functional role, and effective connectivity and probabilistic tractography to map the cerebello-cerebral mentalizing network. We …

Repeated Endo-Tracheal Tube Disconnection Generates Pulmonary Edema In A Model Of Volume Overload: An Experimental Study, Bhushan H Katira, Doreen Engelberts, Sheena Bouch, Jordan Fliss, Luca Bastia, Kohei Osada, Kim A Connelly, Marcelo B P Amato, Niall D Ferguson, Wolfgang M Kuebler, Brian P Kavanagh, Laurent J Brochard, Martin Post

2020-Current year OA Pubs

BACKGROUND: An abrupt lung deflation in rodents results in lung injury through vascular mechanisms. Ventilator disconnections during endo-tracheal suctioning in humans often cause cardio-respiratory instability. Whether repeated disconnections or lung deflations cause lung injury or oedema is not known and was tested here in a porcine large animal model.

METHODS: Yorkshire pigs (~ 12 weeks) were studied in three series. First, we compared PEEP abruptly deflated from 26 cmH

RESULTS: As compared to PEEP 5 cmH

CONCLUSION: Single abrupt deflation from high PEEP, and repeated short deflations from moderate PEEP cause pulmonary oedema, impaired oxygenation, and increased PVR, in this …

Sustained Efficacy And Safety Of Burosumab, A Monoclonal Antibody To Fgf23, In Children With X-Linked Hypophosphatemia, Agnès Linglart, Erik A Imel, Michael P Whyte, Anthony A Portale, Wolfgang Högler, Annemieke M Boot, Raja Padidela, William Van't Hoff, Gary S Gottesman, Angel Chen, Alison Skrinar, Mary Scott Roberts, Thomas O Carpenter

2020-Current year OA Pubs

PURPOSE: In X-linked hypophosphatemia (XLH), excess fibroblast growth factor-23 causes hypophosphatemia and low calcitriol, leading to musculoskeletal disease with clinical consequences. XLH treatment options include conventional oral phosphate with active vitamin D, or monotherapy with burosumab, a monoclonal antibody approved to treat children and adults with XLH. We have previously reported outcomes up to 64 weeks, and here we report safety and efficacy follow-up results up to 160 weeks from an open-label, multicenter, randomized, dose-finding trial of burosumab for 5- to 12-year-old children with XLH.

METHODS: After 1 week of conventional therapy washout, patients were randomized 1:1 to burosumab every …

The Prevalance Of Binge Eating Disorder And Associated Psychiatric And Substance Use Disorders In A Student Population In Kenya - Towards A Public Health Approach, Victoria N Mutiso, David M Ndetei, Esther N Muia, Rita K Alietsi, Lydia Onsinyo, Frida Kameti, Monicah Masake, Christine Musyimi, Daniel Mamah

2020-Current year OA Pubs

INTRODUCTION: Kenya in particular and Africa in general lack data on Binge Eating Disorder (BED). The overarching objective of this study is to fill that gap. Kenyans may not be aware that BED exists when a "very good" appetite is considered a sign of good health, especially if food is available either at home, in fast food shops or when communally eating together, a very common cultural practice. On the other hand where there is relatively insufficient food, it is not expected that one could be having a problem of eating too much.

METHOD: We administered the following tools and …

Relationships Between Correlated Spikes, Oxygen And Lfp In The Resting-State Primate, Jingfeng M Li, Benjamin T Acland, Alexander S Brenner, William J Bentley, Lawrence H Snyder

2020-Current year OA Pubs

Resting-state functional MRI (rsfMRI) provides a view of human brain organization based on correlation patterns of blood oxygen level dependent (BOLD) signals recorded across the whole brain. The neural basis of resting-state BOLD fluctuations and their correlation remains poorly understood. We simultaneously recorded oxygen level, spikes, and local field potential (LFP) at multiple sites in awake, resting monkeys. Following a spike, the average local oxygen and LFP voltage responses each resemble a task-driven BOLD response, with LFP preceding oxygen by 0.5 s. Between sites, features of the long-range correlation patterns of oxygen, LFP, and spikes are similar to features seen …

Therapeutic Efficacy Of 177lu-Labeled A20fmdv2 Peptides Targeting Ανβ6, Truc Thao Huynh, Sreeja Sreekumar, Cedric Mpoy, Buck Edward Rogers

2020-Current year OA Pubs

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Hydraulic Permeability And Compressive Properties Of Porcine And Human Synovium, Milad Rohanifar, Benjamin B Johnston, Alexandra L Davis, Young Guang, Kayla Nommensen, James A J Fitzpatrick, Christine N Pham, Lori A Setton

2020-Current year OA Pubs

The synovium is a multilayer connective tissue separating the intra-articular spaces of the diarthrodial joint from the extra-synovial vascular and lymphatic supply. Synovium regulates drug transport into and out of the joint, yet its material properties remain poorly characterized. Here, we measured the compressive properties (aggregate modulus, Young's modulus, and Poisson's ratio) and hydraulic permeability of synovium with a combined experimental-computational approach. A compressive aggregate modulus and Young's modulus for the solid phase of synovium were quantified from linear regression of the equilibrium confined and unconfined compressive stress upon strain, respectively (H

Filtering Respiratory Motion Artifact From Resting State Fmri Data In Infant And Toddler Populations, Sydney Kaplan, Dominique Meyer, Dimitrios Alexopoulos, Deanna M Barch, Adam T Eggebrecht, Jeanette K Kenley, Cynthia E Rogers, Muriah D Wheelock, Christopher D Smyser, Et Al.

2020-Current year OA Pubs

The importance of motion correction when processing resting state functional magnetic resonance imaging (rs-fMRI) data is well-established in adult cohorts. This includes adjustments based on self-limited, large amplitude subject head motion, as well as factitious rhythmic motion induced by respiration. In adults, such respiration artifact can be effectively removed by applying a notch filter to the motion trace, resulting in higher amounts of data retained after frame censoring (e.g., "scrubbing") and more reliable correlation values. Due to the unique physiological and behavioral characteristics of infants and toddlers, rs-fMRI processing pipelines, including methods to identify and remove colored noise due to …

Enhancing Task Fmri Preprocessing Via Individualized Model-Based Filtering Of Intrinsic Activity Dynamics, Matthew F Singh, Anxu Wang, Michael Cole, Shinung Ching, Todd S Braver

2020-Current year OA Pubs

Brain responses recorded during fMRI are thought to reflect both rapid, stimulus-evoked activity and the propagation of spontaneous activity through brain networks. In the current work, we describe a method to improve the estimation of task-evoked brain activity by first "filtering-out the intrinsic propagation of pre-event activity from the BOLD signal. We do so using Mesoscale Individualized NeuroDynamic (MINDy; Singh et al. 2020b) models built from individualized resting-state data to subtract the propagation of spontaneous activity from the task-fMRI signal (MINDy-based Filtering). After filtering, time-series are analyzed using conventional techniques. Results demonstrate that this simple operation significantly improves the statistical …

Prevention Is Political: Political Party Affiliation Predicts Perceived Risk And Prevention Behaviors For Covid-19, Marc T Kiviniemi, Heather Orom, Jennifer L Hay, Erika A Waters

2020-Current year OA Pubs

BACKGROUND: Many US politicians have provided mixed messages about the risks posed by SARS-CoV-2/COVID-19 and whether and to what extent prevention practices should be put in place to prevent transmission. This politicization of the virus and pandemic may affect individuals' risk perceptions and willingness to take precautions. We examined how political party affiliation relates to risk perception for one's own and other people's likelihood of SARS-CoV-2 infection/COVID-19 illness.

METHODS: We surveyed members of a nationally-representative, probability-sampling based survey panel (N = 410) to examine their risk perceptions, precautionary behaviors, and political party affiliation.

RESULTS: The more strongly one identified as …

Intracranial Internal Carotid Artery Calcification Is Not Predictive Of Future Cognitive Decline, Farzaneh Rahmani, Marina Nguyen, Charles D Chen, Nicole Mckay, Aylin Dincer, Nelly Joseph-Mathurin, Gengsheng Chen, Jingxia Liu, Hilary L P Orlowski, John C Morris, Tammie L S Benzinger

2020-Current year OA Pubs

BACKGROUND: Intracranial internal carotid artery (ICA) calcification is a common incidental finding in non-contrast head CT. We evaluated the predictive value of ICAC (ICAC) for future risk of cognitive decline and compared the results with conventional imaging biomarkers of dementia.

METHODS: In a retrospective observational cohort, we included 230 participants with a PET-CT scan within 18 months of a baseline clinical assessment and longitudinal imaging assessments. Intracranial ICAC was quantified on baseline CT scans using the Agatson calcium score, and the association between baseline ICA calcium scores and the risk of conversion from a CDR of zero in baseline to …

Community-Level Factors Associated With Geographic Access To Food Retailers Offering Nutrition Incentives In Chicago, Illinois, Chelsea R Singleton, Fikriyah Winata, Alexandra M Roehll, Isa Adamu, Gabriella M Mcloughlin

2020-Current year OA Pubs

INTRODUCTION: Nutrition incentive programs provide low-income populations with a monetary resource to make healthy foods affordable and accessible. This study aimed to use geospatial analysis to evaluate availability of the Link Match nutrition incentive program in Chicago, Illinois, to determine whether underresourced communities have access.

METHODS: We obtained 2018 spatial data on census tract-level sociodemographic characteristics in Chicago. Fifty-seven retailers (eg, farmers markets, food cooperatives) offered Link Match across the city's 801 census tracts. We examined ordinary least squares and spatial lag regression models to identify census tract-level variables associated with distance (in miles) from the nearest Link Match retailer. …

Single-Agent Mosunetuzumab Shows Durable Complete Responses In Patients With Relapsed Or Refractory B-Cell Lymphomas: Phase I Dose-Escalation Study, Lihua E Budde, Nancy L Bartlett, Et Al

2020-Current year OA Pubs

PURPOSE: Mosunetuzumab is a bispecific antibody targeting CD20 and CD3 that redirects T cells to engage and eliminate malignant B cells and is being developed for relapsed or refractory (R/R) B-cell non-Hodgkin lymphomas (B-NHLs).

METHODS: This first-in-human trial (ClinicalTrials.gov identifier: NCT02500407) evaluated the safety and tolerability and efficacy of mosunetuzumab in patients with R/R B-NHL and established the recommended phase II dose. Data from dose escalation are presented. Single-agent mosunetuzumab was administered intravenously in 3-week cycles, at full dose in cycle 1 day 1 (group A) or with ascending (step-up) doses during cycle 1 on days 1, 8, and 15 …

Bridging The Gap To Advance The Care Of Individuals With Cancer: Collaboration And Partnership In The Cardiology Oncology Innovation Network (Coin), Sherry-Ann Brown, Craig Beavers, Hugo R Martinez, Catherine H Marshall, Iredia M Olaye, Avirup Guha, David Cho, Alison Bailey, Carmen Bergom, Neha Bansal, Brenton Bauer, Richard K Cheng

2020-Current year OA Pubs

Cardiovascular diseases and cancer continue to be the two leading causes of death in the United States. While innovations in artificial intelligence, digital health, and telemedicine may revolutionize cardio-oncology clinical practice, barriers to widespread adoption continue to exist. The most effective way to advance these technologies is through a broad range of stakeholders sharing a common vision. Additionally, as we enter the digital era in healthcare, we must help lead this charge for the benefit of our cardiology and oncology patients. Bolstering collaborations in cardiology and oncology is key, in partnership with technology firms, industry, academia, and private practice, with …

Disseminated Infection Due To Neocosmospora (Fusarium) Falciformis In A Patient With Acute Myelogenous Leukemia, Ra'ed Jabr, Rachael M Liesman, Venkata Rakesh Sethapati, D Matthew Shoemaker, Andrej Spec, Wissam El Atrouni

2020-Current year OA Pubs

No abstract provided.

Regulation Of The Double-Stranded Rna Response Through Adar1 Licenses Metaplastic Reprogramming In Gastric Epithelium, José B. Sáenz, Nancy Vargas, Charles J. Cho, Jason C. Mills

2020-Current year OA Pubs

Cells recognize both foreign and host-derived double-stranded RNA (dsRNA) via a signaling pathway that is usually studied in the context of viral infection. It has become increasingly clear that the sensing and handling of endogenous dsRNA is also critical for cellular differentiation and development. The adenosine RNA deaminase, ADAR1, has been implicated as a central regulator of the dsRNA response, but how regulation of the dsRNA response might mediate cell fate during injury and whether such signaling is cell intrinsic remain unclear. Here, we show that the ADAR1-mediated response to dsRNA was dramatically induced in 2 distinct injury models of …

Videos Of Sipuleucel-T Programmed T Cells Lysing Cells That Express Prostate Cancer Target Antigens, Adam S Kibel, Brant A Inman, Russell K Pachynski, Tuyen Vu, Nadeem A Sheikh, Daniel P Petrylak

2020-Current year OA Pubs

Sipuleucel-T, an autologous cellular immunotherapy, was approved to treat metastatic castration-resistant prostate cancer in 2010 in the United States. Treatment with sipuleucel-T primes the immune system to target prostate acid phosphatase, which is expressed by prostate cancer cells, potentially leading to lysis of cancer cells. Expanding on previously reported indirect evidence of cell killing with sipuleucel-T treatment, we sought to provide direct evidence of cell lysis through visualization. We used advanced video technology and available samples of peripheral blood mononuclear cells from subjects enrolled in the STAMP trial (NCT01487863). Isolated CD8+ T cells were used as effector cells and cocultured …

Highly Reliable Creation Of Floxed Alleles By Electroporating Single-Cell Embryos, Monica F Sentmanat, J Michael White, Evguenia Kouranova, Xiaoxia Cui

2020-Current year OA Pubs

BACKGROUND: Floxed (flanked by loxP) alleles are a crucial portion of conditional knockout mouse models. However, an efficient and reliable strategy to flox genomic regions of any desired size is still lacking.

RESULTS: Here, we demonstrate that the method combining electroporation of fertilized eggs with gRNA/Cas9 complexes and single-stranded oligodeoxynucleotides (ssODNs), assessing phasing of loxP insertions in founders using an in vitro Cre assay and an optional, highly specific and efficient second-round targeting ensures the generation of floxed F1 animals in roughly five months for a wide range of sequence lengths (448 bp to 160 kb reported here).

CONCLUSIONS: Floxed …