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Simultaneous Gene Excision and Integration by Dual-Guide CRISPR-Cas9.

Methods Mol Biol 2021 ;2307:69-83

Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA.

Metabolic engineering frequently requires both gene knockouts and gene integration. CRISPR-Cas9 has been extensively used to create double-stranded DNA breaks that result in indel mutations; however, such mutations can revert or create toxic product. Gene integration can also be accomplished by CRISPR-Cas9 introduced double-stranded DNA breaks and a donor DNA cassette. Read More

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January 2021

Single-Stranded DNA Curtains for Single-Molecule Visualization of Rad51-ssDNA Filament Dynamics.

Methods Mol Biol 2021 ;2281:193-207

Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.

Homologous recombination (HR) is a highly conserved DNA repair pathway required for the accurate repair of DNA double-stranded breaks. DNA recombination is catalyzed by the RecA/Rad51 family of proteins, which are conserved from bacteria to humans. The key intermediate catalyzing DNA recombination is the presynaptic complex (PSC), which is a helical filament comprised of Rad51-bound single-stranded DNA (ssDNA). Read More

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January 2021

Generation of Fluorescent Versions of Saccharomyces cerevisiae RPA to Study the Conformational Dynamics of Its ssDNA-Binding Domains.

Methods Mol Biol 2021 ;2281:151-168

Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO, USA.

Replication protein A (RPA) is an essential single-stranded DNA (ssDNA)-binding protein that sequesters ssDNA and protects it from nucleolytic degradation. The RPA-ssDNA nucleoprotein acts as a hub to recruit over two dozen DNA metabolic enzymes onto ssDNA to coordinate DNA replication, repair, and recombination. RPA functions as a heterotrimer composed of RPA70, RPA32, and RPA14 subunits and has multiple DNA-binding and protein-interaction domains. Read More

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January 2021

Dual recombinases-based genetic lineage tracing for stem cell research with enhanced precision.

Sci China Life Sci 2021 Apr 9. Epub 2021 Apr 9.

State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.

Stem cell research has become a hot topic in biology, as the understanding of stem cell biology can provide new insights for both regenerative medicine and clinical treatment of diseases. Accurately deciphering the fate of stem cells is the basis for understanding the mechanism and function of stem cells during tissue repair and regeneration. Cre-loxP-mediated recombination has been widely applied in fate mapping of stem cells for many years. Read More

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Traumatic physeal fractures in cats: a review of 36 cases (2010-2020).

J Feline Med Surg 2021 Apr 13:1098612X211005886. Epub 2021 Apr 13.

Department of Clinical Science and Services, Queen Mother Hospital for Animals, The Royal Veterinary College, Hatfield, UK.

Objectives: The aim of this study was to describe the demography, aetiology, location and classification of physeal fractures in cats, and to describe their management and outcomes.

Methods: Clinical records and radiographs of cats referred for management of physeal fractures were retrospectively reviewed. Fractures of the proximal femoral physis were excluded. Read More

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A case of Spigelian hernia after laparoscopic incisional hernia repair.

Asian J Endosc Surg 2021 Apr 13. Epub 2021 Apr 13.

Department of Surgery, Hyogo Prefectural Kakogawa Medical Center, Kakogawa, Japan.

Laparoscopic ventral hernia repair with intraperitoneal onlay mesh reinforcement is often performed in clinical practice. We herein describe a patient who developed a Spigelian hernia at the edge of the mesh due to rupture of the muscular layer in the abdominal wall. A 69-year-old woman developed a left-sided abdominal bulge 15 months after laparoscopic ventral hernia repair. Read More

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Urethral plate quality assessment and its impact on hypospadias repair outcomes: A systematic review and quality assessment.

J Pediatr Urol 2021 Feb 23. Epub 2021 Feb 23.

Pediatric Urology Division, Surgery Department, Sidra Medicine, Doha, Qatar.

Background: The variability of the urethral plate (UP) characteristics is one of the factors that influence technical choices for hypospadias correction. However, it is difficult to objectively evaluate the UP, leading to controversies in this subject, and vague terms utilized in the literature to describe its characteristics.

Objective: We aim to analyze the previously described methods used to characterize and evaluate UP quality, emphasizing the pros and cons of each system, and highlighting its possible influence on different postoperative outcomes. Read More

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February 2021

A Systematic Review Demonstrates High Variation of Terminology, Surgical Techniques, Preoperative Diagnostic Measures, and Geographic Differences in the Treatment of Athletic Pubalgia/Sports Hernia/Core Muscle Injury/Inguinal Disruption.

Arthroscopy 2021 Apr 9. Epub 2021 Apr 9.

St. Joseph's University Medical Center, Department of Orthopaedic Surgery, Paterson NJ 07503; New Jersey Orthopaedic Institute, Wayne NJ 07470.

Purpose: To perform a systematic review of reported terminologies, surgical techniques, preoperative diagnostic measures, and geographic differences in the treatment of core muscle injury (CMI)/athletic pubalgia/inguinal disruption.

Methods: A systematic review was performed by searching PubMed, Cochrane Library, and Embase to identify clinical studies or articles that described a surgical technique to treat CMI refractory to nonoperative treatment. The search phrase used was "core muscle injury" OR "sports hernia" OR "athletic pubalgia" OR "inguinal disruption". Read More

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Building the alliance and using experiential techniques in the early phases of psychotherapy for avoidant personality disorder.

J Clin Psychol 2021 Apr 12. Epub 2021 Apr 12.

Center for Metacognitive Interpersonal Therapy, Rome, Italy.

Individuals experiencing avoidant personality disorder (AvPD) tend to make sense of social interactions via maladaptive self-and other attributions. They also experience difficulties in recognizing emotions. A further feature of AvPD psychopathology is the tendency to resort to maladaptive coping strategies, such as behavioral avoidance and perfectionism. Read More

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Comprehensive Pan-Cancer Analysis Confirmed That ATG5 Promoted the Maintenance of Tumor Metabolism and the Occurrence of Tumor Immune Escape.

Front Oncol 2021 25;11:652211. Epub 2021 Mar 25.

School of Medicine, Shandong University, Jinan, China.

Background: Autophagy related protein 5 (ATG5) is an important autophagosome formation related protein, and its involvement in the biological process of autophagy has been shown to correlate with tumor metabolic patterns and the formation of tumor heterogeneity. However, the role of ATG5 in tumor metabolism and tumor immunity remains unclear.

Method: In order to explore this problem, this study was designed to reveal the role of ATG5 in tumor metabolism and tumor immunity through pan-cancer analysis of multi-database. Read More

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Deployment of a bioabsorbable plate as the rigid buttress for skull base repair after endoscopic pituitary surgery.

Gland Surg 2021 Mar;10(3):1010-1017

Department of Neurosurgery, Qilu Hospital of Shandong University, Jinan, China.

Background: Bioresorbable alloplastic implants have become desirable as a rigid buttress for reconstructing skull base defects. This study aimed to describe the use of a biodegradable plate (PolyMax RAPID) in skull base repair of endoscopic endonasal pituitary surgery and to investigate the clinical outcome and safety of this novel method.

Methods: Between January 2019 and January 2020, 22 patients with pituitary adenomas who underwent endoscopic skull base repair with a Polymax RAPID plate were included. Read More

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Adult pectus excavatum repair: national outcomes of the Nuss and Ravitch procedures.

J Thorac Dis 2021 Mar;13(3):1396-1402

Department of Surgery, The University of Kansas, Kansas City, KS, USA.

Background: National data is limited on pectus excavatum, the most common chest wall deformity which is often repaired using the Ravitch and Nuss procedures. The purpose of the study was to describe demographics and outcomes of adult patients who underwent surgical repair of pectus excavatum via open and minimally invasive thoracoscopic methods.

Methods: A retrospective analysis of the American College of Surgeons National Surgical Quality Improvement Program (ACS NSQIP) database from 2015 to 2018 was performed, capturing patients 18 years or older with pectus excavatum as the postoperative diagnosis. Read More

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Past, present, and perspectives of protein N-terminal methylation.

Curr Opin Chem Biol 2021 Apr 8;63:115-122. Epub 2021 Apr 8.

Department of Medicinal Chemistry and Molecular Pharmacology, Purdue Institute for Drug Discovery, Purdue University Center for Cancer Research, Purdue University, 720 Clinic Drive, West Lafayette, IN, 47907, United States. Electronic address:

The posttranslational methylation of the α-N-terminal amino group of proteins was first documented over 40 years ago, but the functional significance of this modification has been underexplored relative to lysine and arginine methylation. Increasing reports implicates α-N-terminal methylation as a widespread and critical regulator of mitosis, chromatin interactions, DNA repair, and translation fidelity. Here, we summarize advances in the current understanding of protein α-N-terminal methylation biological functions and mechanisms across eukaryotic organisms. Read More

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Isolation and culture of adult murine cardiac atrial and ventricular fibroblasts and myofibroblasts.

Methods 2021 Apr 7. Epub 2021 Apr 7.

Montreal Heart Institute, 5000 Belanger, Montréal, Québec, H1T 1C8, Canada; Departments of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Québec, H3C 3J7, Canada; Pharmacology and Physiology, Université de Montréal, Montréal, Québec, H3C 3J7, Canada; Université de Montréal, Montréal, Québec, H3C 3J7, Canada. Electronic address:

Cardiac fibroblasts play a critical role in extracellular matrix homeostasis, wound healing, and cardiac interstitial fibrosis: the latter being a pathophysiological response to a chronic increase in afterload. Using a standard protocol to isolate cardiac fibroblasts and maintain them in their quiescent phenotype in vitro will enable a better understanding of cardiac fibroblast biology and their role in the response to profibrotic stimuli. Here, we describe an enzymatic method for isolating cardiac fibroblasts. Read More

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Targets identified from exercised heart: killing multiple birds with one stone.

NPJ Regen Med 2021 Apr 9;6(1):23. Epub 2021 Apr 9.

Institute of Geriatrics (Shanghai University), Affiliated Nantong Hospital of Shanghai University (The Sixth People's Hospital of Nantong), School of Medicine, Shanghai University, Nantong, China.

Cardiovascular diseases (CVDs) are a major cause of mortality worldwide, which are mainly driven by factors such as aging, sedentary lifestyle, and excess alcohol use. Exercise targets several molecules and protects hearts against many of these physiological and pathological stimuli. Accordingly, it is widely recognized as an effective therapeutic strategy for CVD. Read More

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Aortic rupture by an acute dissection of the thoracoabdominal aorta in a 12-year-old boy.

Ann Vasc Surg 2021 Apr 6. Epub 2021 Apr 6.

Physician in the Angiology and Vascular Surgery Department, San Carlos Clinical Hospital, Madrid.

Aortic dissection is exceptional in children. When they are affected, many clinical conditions as Marfan syndrome or Ehlers-Danlos syndrome are usually involved, but up to 22% have no associated background. Its high morbidity and low incidence in this age group require a high level of suspicion in children without predisposing factors when the presentation is atypical, to avoid the severe consequences of the delay of their diagnosis. Read More

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Unique Complications and Failure Modes of Iliac Branch Devices.

Ann Vasc Surg 2021 Apr 6. Epub 2021 Apr 6.

Department of Surgery, Division of Vascular & Endovascular Surgery, Stanford University School of Medicine, Stanford, CA.

Objectives: Iliac branch devices (IBDs) are modular, bifurcated endografts designed to preserve hypogastric flow during endovascular aortoiliac aneurysm repair. We report our single center outcomes, and describe the unique complications of these devices: inability to cannulate the hypogastric artery (technical failure), occlusion of the internal branch, and type III endoleak from separation of components between the main body and IBD.

Methods: A prospectively maintained institutional database of patients undergoing IBD implantation between 2014 and 2019 was reviewed. Read More

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