Boundary stacking interactions enable cross-TAD enhancer-promoter communication during limb development.

Although promoters and their enhancers are frequently contained within a topologically associating domain (TAD), some developmentally important genes have their promoter and enhancers within different TADs. Hypotheses about molecular mechanisms enabling cross-TAD interactions remain to be assessed. To test these hypotheses, we used optical reconstruction of chromatin architecture to characterize the conformations of the Pitx1 locus on single chromosomes in developing mouse limbs. Our data support a model in which neighboring boundaries are stacked as a result of loop extrusion, bringing boundary-proximal cis-elements into contact. This stacking interaction also contributes to the appearance of architectural stripes in the population average maps. Through molecular dynamics simulations, we found that increasing boundary strengths facilitates the formation of the stacked boundary conformation, counter-intuitively facilitating border bypass. This work provides a revised view of the TAD borders' function, both facilitating and preventing cis-regulatory interactions, and introduces a framework to distinguish border-crossing from border-respecting enhancer-promoter pairs.

Construction of intracellular asymmetry and asymmetric division in Escherichia coli.

The design principle of establishing an intracellular protein gradient for asymmetric cell division is a long-standing fundamental question. While the major molecular players and their interactions have been elucidated via genetic approaches, the diversity and redundancy of natural systems complicate the extraction of critical underlying features. Here, we take a synthetic cell biology approach to construct intracellular asymmetry and asymmetric division in Escherichia coli, in which division is normally symmetric. We demonstrate that the oligomeric PopZ from Caulobacter crescentus can serve as a robust polarized scaffold to functionalize RNA polymerase. Furthermore, by using another oligomeric pole-targeting DivIVA from Bacillus subtilis, the newly synthesized protein can be constrained to further establish intracellular asymmetry, leading to asymmetric division and differentiation. Our findings suggest that the coupled oligomerization and restriction in diffusion may be a strategy for generating a spatial gradient for asymmetric cell division.

Electrodeposition of Silver in a Ternary Deep Eutectic Solvent and the Electrochemical Sensing Ability of the Ag-Modified Electrode for Nitrofurazone.

The determination of nitrofurazone (NFZ) has received substantial attention because it is a kind of antibiotic drug. Herein, a rapid and low-cost electrochemical sensor for the analysis of NFZ is reported. The method uses Ag-modified electrodes in which different surfactants, hexadecyltrimethylammonium bromide and sodium dodecyl sulfate, in a ternary choline chloride-urea-glycerol deep eutectic solvent were deposited. The physical properties of the solutions with various surfactants are investigated by a conductivity meter, viscometer, and tensiometer. The morphologies and crystallinity of the Ag-modified electrodes were characterized by using scanning electron microscopy, transmission electron microscopy, and powder X-ray diffraction. Electrochemical impedance spectroscopy and CV analyses indicate that the as-prepared Ag-SDS electrode exhibited better performance as a NFZ sensor. The dynamic linear range of NFZ is 0.66-930 µM with a corresponding detection limit of 0.37 µM. The proposed electrochemical sensor was applied to detect NFZ in the aquaculture water sample, and the results showed good recovery in the range from 100.28 to 102.65%.

Selective Targeting of Vibrios by Fluorescent Siderophore-Based Probes.

Siderophores are small molecules used to specifically transport iron into bacteria via related receptors. By adapting siderophores and hijacking their pathways, we may discover an efficient and selective way to target microbes. Herein, we report the synthesis of a siderophore-fluorophore conjugate VF-FL derived from vibrioferrin (VF). Using flow cytometry and fluorescence microscopy, the probe selectively labeled vibrios, including V. parahaemolyticus, V. cholerae, and V. vulnificus, even in the presence of other species such as S. aureus and E. coli. The labeling is siderophore-related and both iron-limited conditions and the siderophore moiety are required. The competitive relationship between VF-FL and VF in vibrios implies an unreported VF-related transport mechanism in V. cholerae and V. vulnificus. These studies demonstrate that the siderophore scaffold provides a method to selectively target microbes expressing cognate receptors under iron-limited conditions.

Effectiveness of Crown Preparation Assessment Software As an Educational Tool in Simulation Clinic: A Pilot Study.

The aim of this pilot study was to evaluate the feasibility of a new tooth preparation assessment software, Preppr, as an educational tool for dental students in achieving optimal parameters for a crown preparation. In February 2015, 30 dental students in their fourth year in a five-year undergraduate dental curriculum in New Zealand were randomly selected from a pool of volunteers (N=40) out of the total class of 85. The participants were placed into one of three groups of ten students each: Group A, the control group, received only written and pictorial instructions; Group B received tutor evaluation and feedback; and Group C performed self-directed learning with the aid of Preppr. Each student was asked to prepare an all-ceramic crown on the lower first molar typodont within three hours and to repeat the exercise three times over the next four weeks. The exercise stipulated a 1 mm finish line dimension and total convergence angles (TOC) between 10 and 20 degrees. Fulfillment of these parameters was taken as an acceptable preparation. The results showed that Group C had the highest percentage of students who achieved minimum finish line dimensions and acceptable TOC angles. Those students also achieved the stipulated requirements earlier than the other groups. This study's findings provide promising data on the feasibility of using Preppr as a self-directed educational tool for students training to prepare dental crowns.

The Species and Origin of Shark Fins in Taiwan's Fishing Ports, Markets, and Customs Detention: A DNA Barcoding Analysis.

The increasing consumption of shark products, along with the shark's fishing vulnerabilities, has led to the decrease in certain shark populations. In this study we used a DNA barcoding method to identify the species of shark landings at fishing ports, shark fin products in retail stores, and shark fins detained by Taiwan customs. In total we identified 23, 24, and 14 species from 231 fishing landings, 316 fin products, and 113 detained shark fins, respectively. All the three sample sources were dominated by Prionace glauca, which accounted for more than 30% of the collected samples. Over 60% of the species identified in the fin products also appeared in the port landings, suggesting the domestic-dominance of shark fin products in Taiwan. However, international trade also contributes a certain proportion of the fin product markets, as four species identified from the shark fin products are not found in Taiwan's waters, and some domestic-available species were also found in the customs-detained sample. In addition to the species identification, we also found geographical differentiation in the cox1 gene of the common thresher sharks (Alopias vulpinus), the pelagic thresher shark (A. pelagicus), the smooth hammerhead shark (Sphyrna zygaena), and the scalloped hammerhead shark (S. lewini). This result might allow fishing authorities to more effectively trace the origins as well as enforce the management and conservation of these sharks.