Intrinsic deletion at 10q23.31, including the gene locus, is aggravated upon CRISPR-Cas9-mediated genome engineering in HAP1 cells mimicking cancer profiles.

The CRISPR-Cas9 system is a powerful tool for studying gene functions and holds potential for disease treatment. However, precise genome editing requires thorough assessments to minimize unintended on- and off-target effects. Here, we report an unexpected 283-kb deletion on Chromosome 10 (10q23.

Targeted Muscle Reinnervation to Reduce Postoperative Pain in Patients Undergoing Hindquarter Resections.

Background/aim: Pre-emptive targeted muscle reinnervation (TMR) at the time of amputation results in less phantom limb pain (PLP) compared with untreated amputee controls. There is limited literature describing the technique in patients undergoing hindquarter amputation despite up to 90% of these patients reporting PLP and 50% presenting with painful neuroma. The purpose of the current study was to describe the motor nerves accessible through a primary hind-quarter amputation to be used for TMR and review pain outcomes in clinical case correlates of patients with TMR.

Data quality and auditing within the Netherlands Heart Registration: using the PCI registry as an example.

Aim: The aim of this article is to present the method and results of the data quality control system and audit within the Netherlands Heart Registration (NHR) using data of patients treated with percutaneous coronary intervention (PCI) in the Netherlands as an example.

Methods: The NHR is a Dutch nationwide registry of all cardiac interventions, comprising data from all 71 hospitals, of which 30 are cardiac intervention or heart centres. Each year, within the NHR, data validation and verification is performed by standard quality controls and monitoring visits (audits).

Protocol: A Multiplexed Reporter Assay to Study Effects of Chromatin Context on DNA Double-Strand Break Repair.

DNA double-strand breaks (DSBs) can be repaired through various pathways. Understanding how these pathways are regulated is of great interest for cancer research and optimization of gene editing. The local chromatin environment can affect the balance between repair pathways, but this is still poorly understood.

Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance.

DNA double-strand break (DSB) repair is mediated by multiple pathways. It is thought that the local chromatin context affects the pathway choice, but the underlying principles are poorly understood. Using a multiplexed reporter assay in combination with Cas9 cutting, we systematically measure the relative activities of three DSB repair pathways as a function of chromatin context in >1,000 genomic locations.

Multi-Institutional Prospective Randomized Control Trial of Novel Intracorporeal Lithotripters: ShockPulse-SE Trilogy Trial.

Currently, there are multiple intracorporeal lithotripters available for use in percutaneous nephrolithotomy (PCNL). This study aimed to evaluate the efficiency of two novel lithotripters: Trilogy and ShockPulse-SE. This is a prospective multi-institutional randomized trial comparing outcomes of PCNL using two novel lithotripters between February 2019 and June 2020.

Diaphragmatic dysfunction in dogs with cervical spinal disorders before and after surgery using fluoroscopy, motion-mode ultrasound and radiography was not different than a group of control dogs.

Cervical spinal disorders can lead to life-threatening respiratory complications. Diaphragmatic dysfunction is attributed to spinal cord morbidity secondary to cervical myelopathy or decompressive surgical intervention. The purpose of this observational case-control study was to determine the frequency of diaphragmatic dysfunction in dogs with cervical spinal disorders and a control group, the strength of association between cervical myelopathies and decompressive surgery with diaphragm paresis, and the agreement between and clinical usefulness of fluoroscopy, motion-mode ultrasonography, and radiography for diagnosing diaphragmatic dysfunction.

Soil functional responses to drought under range-expanding and native plant communities.

Current climate warming enables plant species and soil organisms to expand their range to higher latitudes and altitudes. At the same time, climate change increases the incidence of extreme weather events such as drought. While it is expected that plants and soil organisms originating from the south are better able to cope with drought, little is known about the consequences of their range shifts on soil functioning under drought events.

Association of meniscal injury to joint space width on standard tibial plateau leveling osteotomy lateral radiographic projections of the canine stifle.

Up to 70% of dogs with cranial cruciate ligament tears have concurrent meniscal injury, and these injuries can increase the risk of developing osteoarthritis and persistent lameness. Studies assessing joint space width on knee radiographs in people have indicated associations between joint space width and meniscal injuries. The aim of this prospective analytical study was to determine if there was an association between stifle joint space width on three different radiographic projections (the standard tibial plateau leveling osteotomy projections and a standing lateral projection) and meniscal injuries identified at surgery in dogs.

Effects of short- and long-term administration of nonsteroidal anti-inflammatory drugs on osteotomy healing in dogs.

Objective: To determine the influence of short-term administration of carprofen on bone healing in dogs.

Study Design: Randomized controlled experimental study.

Animals: Eighteen purpose-bred sexually mature hound dogs.

Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDER.

Current genome editing tools enable targeted mutagenesis of selected DNA sequences in many species. However, the efficiency and the type of introduced mutations by the genome editing method are largely dependent on the target site. As a consequence, the outcome of the editing operation is difficult to predict.

Fin healing and regeneration in sturgeon.

Pectoral fin healing in fin spines and rays were examined in juvenile Atlantic sturgeon Acipenser oxyrinchus oxyrinchus following three different sampling techniques (n = 8-9 fish per treatment): entire leading fin spine removed, a 1-2 cm portion removed near the point of articulation, or a 1-2 cm portion removed from a secondary fin ray. Also, to determine whether antibiotic treatment influences healing, an additional group of fish (n = 8) was not given an injection of an oxytetracycline (OTC)-based antibiotic following removal of the entire leading fin spine. Following fin sampling, fish from different treatments were mixed equally between three large (4,000 I) recirculating systems and fin-ray healing and mortality were monitored over a 12 month period.

Mapping 3D genome organization relative to nuclear compartments using TSA-Seq as a cytological ruler.

While nuclear compartmentalization is an essential feature of three-dimensional genome organization, no genomic method exists for measuring chromosome distances to defined nuclear structures. In this study, we describe TSA-Seq, a new mapping method capable of providing a "cytological ruler" for estimating mean chromosomal distances from nuclear speckles genome-wide and for predicting several Mbp chromosome trajectories between nuclear compartments without sophisticated computational modeling. Ensemble-averaged results in K562 cells reveal a clear nuclear lamina to speckle axis correlated with a striking spatial gradient in genome activity.

Kinetics and Fidelity of the Repair of Cas9-Induced Double-Strand DNA Breaks.

The RNA-guided DNA endonuclease Cas9 is a powerful tool for genome editing. Little is known about the kinetics and fidelity of the double-strand break (DSB) repair process that follows a Cas9 cutting event in living cells. Here, we developed a strategy to measure the kinetics of DSB repair for single loci in human cells.

Easy quantification of template-directed CRISPR/Cas9 editing.

Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE method for quantification of templated editing events, including point mutations.

Changing soil legacies to direct restoration of plant communities.

It is increasingly acknowledged that soil biota may influence interactions among plant species; however, little is known about how to change historical influences of previous land management on soil biota, the so-called 'biotic soil legacy effect'. We used a two-phase plant community-soil feedback approach to study how plant species typical to original (i.e.