Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System

The methodology for site-directed editing of single nucleotides in the vertebrate genome is of considerable interest for research in biology and medicine. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 type II (Cas9) system has emerged as a simple and inexpensive tool for editing genomic loci of interest in a variety of animal models. In zebrafish, error-prone non-homologous end joining (NHEJ) has been used as a simple method to disrupt gene function. We sought to develop a method to easily create site-specific SNPs in the zebrafish genome. Here, we report simple methodologies for using CRISPR/Cas9-mediated homology directed repair using single-stranded oligodeoxynucleotide donor templates (ssODN) for site-directed single nucleotide editing, for the first time in two disease-related genes, tardbp and fus.     

Kinetic Curve Type Assessment for Classification of Breast Lesions Using Dynamic Contrast-Enhanced MR Imaging

ObjectiveThe aim of this study was to employ a kinetic model with dynamic contrast enhancement-magnetic resonance imaging to develop an approach that can efficiently distinguish malignant from benign lesions.ResultsAn average sensitivity of 82%, a specificity of 65%, an area under the receiver operating characteristic curve of 0.76, and a positive predictive value of 82% and negative predictive value of 63% was shown with the kinetic model (p = 0.017, 0.052, 0.068), as compared to an average sensitivity of 80%, a specificity of 55%, an area under the receiver operating characteristic of 0.69, and a positive predictive value of 79% and negative predictive value of 57% with the time-signal intensity curve method (p = 0.003, 0.004, 0.008). The diagnostic consistency of the three radiologists was shown by the κ-value, 0.857 (p<0.001) with the method based on the time-signal intensity curve and 0.826 (p<0.001) with the method of the kinetic model.ConclusionsAccording to the statistic results based on the 46 lesions, the kinetic modeling curve method showed higher sensitivity, specificity, positive and negative predictive values as compared with the time-signal intensity curve method in lesion classification.     

Remediation of Nitrogen-Contaminated Sediment Using Bioreactive,Thin-layer Capping with Biozeolite

In situ amendment of nitrogen-contaminated sediment using bioreactive, thin-layer capping (BTC) with biozeolite (i.e., zeolite with heterotrophic nitrifiers as well as aerobic denitrifiers attached) was studied herein. BTC with biozeolite for nitrogen-contaminated sediment management was evaluated through long-term (170 days) sediment incubation laboratory experiments. The results showed that BTC with relatively small dose rates (<10 kg m^?2) of biozeolite reduced the total nitrogen (TN) concentration in overlying water by over 90%, so it was effective in reducing the amount of N released from sediment. Higher-dose rates of biozeolite capping achieved an even higher removal efficiency. With the DO concentration of 1.5 ～ 6.5 mg L^?1 in overlying water, the reduction efficiency of TN in overlying water using BTC was higher than that less than 1 mg L^?1. In BTC systems, biological regeneration (i.e., heterotrophic nitrifiers attached to zeolite can regenerate the zeolite ion exchange capacity for ammonium) occurred in biozeolite which was saturated with ammonium during the nitrification period. In addition, TN contents in surface sediment in BTC systems were reduced at different levels after the experiment. These findings indicate that the BTC can be a feasible remedial approach to reduce N in overlying water and sediment in eutrophic water bodies. In the BTC, N load was reduced by the added biozeolite through adsorbing ammonium (NH₄⁺-N), converting NH₄⁺-N into nitrate nitrogen (NO₃^?-N) and nitrogen gas (N₂), and assimilating inorganic nitrogen.