Tilleyhooper4996

The CRISPR/Cas9 genome editing tool has the potential to improve the livestock breeding industry by allowing for the introduction of desirable traits. Although an efficient and targeted tool, the CRISPR/Cas9 system can have some drawbacks, including off-target mutations and mosaicism, particularly when used in developing embryos. Here, we introduced genome editing reagents into single-cell bovine embryos to compare the effect of Cas9 mRNA and protein on the mutation efficiency, level of mosaicism, and evaluate potential off-target mutations utilizing next generation sequencing. We designed guide-RNAs targeting three loci (POLLED, H11, and ZFX) in the bovine genome and saw a significantly higher rate of mutation in embryos injected with Cas9 protein (84.2%) vs. Cas9 mRNA (68.5%). In addition, the level of mosaicism was higher in embryos injected with Cas9 mRNA (100%) compared to those injected with Cas9 protein (94.2%), with little to no unintended off-target mutations detected. This study demonstrated that the use of gRNA/Cas9 ribonucleoprotein complex resulted in a high editing efficiency at three different loci in bovine embryos and decreased levels of mosaicism relative to Cas9 mRNA. Additional optimization will be required to further reduce mosaicism to levels that make single-step embryo editing in cattle commercially feasible.Capacitive-resistive electric transfer therapy is used in physical rehabilitation and sports medicine to treat muscle, bone, ligament and tendon injuries. The purpose is to analyze the temperature change and transmission of electric current in superficial and deep knee tissues when applying different protocols of capacitive-resistive electric transfer therapy. Five fresh frozen cadavers (10 legs) were included in this study. Four interventions (high/low power) were performed for 5 min by a physiotherapist with experience. Dynamic movements were performed to the posterior region of the knee. Capsular, intra-articular and superficial temperature were recorded at 1-min intervals and 5 min after the treatment, using thermocouples placed with ultrasound guidance. The low-power protocols had only slight capsular and intra-capsular thermal effects, but electric current flow was observed. The high-power protocols achieved a greater increase in capsular and intra-articular temperature and a greater current flow than the low-power protocols. The information obtained in this in vitro study could serve as basic science data to hypothesize capsular and intra-articular knee recovery in living subjects. The current flow without increasing the temperature in inflammatory processes and increasing the temperature of the tissues in chronic processes with capacitive-resistive electric transfer therapy could be useful for real patients.It is of great significance to study the porosity and permeability properties of medium and low-rank coal. The porosity and permeability in confining stress experiments were used to simulate the porosity and permeability variations of coal samples under different depth conditions. The pore structure of Baoqing coal samples is greatly affected by the confining pressure, and the pores and micro cracks are more easily compressed. Based on the experimental data of mercury intrusion porosimetry (MIP) and nitrogen adsorption (NA), the pore size distributions (PSDs) of medium and low-rank coals were studied. High mercury intrusion pressure would lead to coal matrix compression. Therefore, the pore volume calculated by MIP data was corrected by NA data. The PSDs characteristics of Jixi (JX) coal and Baoqing (BQ) coal samples are obtained from the revised pore volume, and the dominant pores of medium and low-rank coals are obtained. The results show that JX coal has higher spatial heterogeneity, connectivity and pore autocorrelation. Micro fractures have an influence on the autocorrelation and heterogeneity of coal samples, especially for BQ coal samples.Basal leaf angle distribution was surveyed in twenty-one Johnsongrass cultivars near the end of the vegetative stage. The angles increased from the top to the bottom leaves, and compared to cultivated grain sorghums, the average angle was larger in Johnsongrass. When basal leaf angle distribution data were correlated with pathogenicity test data from excised-leaf assays for three isolates of Colletotrichum sublineola, the results showed a weak positive correlation between basal leaf angle and pathogenicity level in Johnsongrass. In order to investigate a protective role of leaf thickness to C. sublineola, leaf thickness was measured in three sorghum cultivars and one Johnsongrass cultivar at the 8-leaf-stage. AZD9291 order Leaf thickness near the apex, near the base, and half-way between the two points were measured in the top four leaves of each plant. Thickness of leaf blade and midrib were recorded separately. Using an excised-leaf-assay, the three points were inoculated with C. sublineola, and pathogenicity level was recorded 4-days-post-inoculation. Results showed strong negative correlations between leaf midrib thickness and pathogenicity level in sorghum and Johnsongrass but not in leaf blades.This work reports a multispectral tomography technique in transmission mode (called 3DITI for 3D Infrared Thermospectroscopic Imaging) based on a middle wavelength infrared (MWIR) focal plane array. This technique relies on an MWIR camera (1.5 to 5.5 μm) used in combination with a multispectral IR monochromator (400 nm to 20 μm), and a sample mounted on a rotary stage for the measurement of its transmittance at several angular positions. Based on the projections expressed in terms of a sinogram, spatial three-dimensional (3D) cubes (proper emission and absorptivity) are reconstructed using a back-projection method based on inverse Radon transform. As a validation case, IR absorptivity tomography of a reflective metallic screw is performed within a very short time, i.e., shorter than 1 min, to monitor 72 angular positions of the sample. Then, the absorptivity and proper emission tomographies of a butane-propane-air burner flame and microfluidic perfluoroalkoxy (PFA) tubing filled with water and ethanol are obtained.