DataSheet_1_Field determination of nitrate in seawater using a novel on-line coppered cadmium column: A comparison study with the vanadium reduction method.docx

Nitrate is the main form of dissolved inorganic nitrogen, playing an important role in both marine biogeochemical research and water environment management. In this work, the most commonly used coppered cadmium column was modified and a novel on-line Cu/Cd column with a spiral structure coupled with a de-bubbling device and syringe-type filter was developed. With the advantages of convenience, portability, stability, and high reduction efficiency, the interference of air bubbles in the column could easily be avoided. Based on the classic Griess reaction, a simple reverse flow injection system coupled with a novel Cu/Cd column and custom-made flow cell was established for the field spectrophotometric determination of nitrate in seawater. The effects of certain reaction parameters—including the reagent concentration, flow rate, length of the Cu/Cd column, and salinity—were investigated, optimized, and compared with pure water, with an approximate 9% increase in the sensitivity of seawater samples. This method exhibited a detection limit of 0.03 μmol/L, with a relative standard deviation of 0.6%, and the working range was 20 μmol/L before dilution. Compared to the referred vanadium reduction method based on the same flow system, the proposed method showed significant advantages including sensitivity and reproducibility. No significant difference was observed between the analytical seawater sample results obtained by the proposed and reference methods. Furthermore, the proposed method was validated by the first class of National Certified Reference Materials and successfully applied to the nitrate determination of Wenling coastal seawater (Zhejiang, China).</p

Multiplexed genome engineering for porcine fetal fibroblasts with gRNA–tRNA arrays based on CRISPR/Cas9

Multiplex gene modifications are highly required for various fields of porcine research. In many species, the CRISPR/Cas9 system has been widely applied for genomic editing and provides a potential tool for introducing multiplex genome mutations simultaneously. Here, we present a CRISPR–Cas9 gRNA–tRNA array (GTR–CRISPR) for multiplexed engineering of porcine fetal fibroblasts (PFFs). We successfully produced multiple sgRNAs using only one Pol III promoter by taking advantage of the endogenous tRNA processing mechanism in porcine cells. Using an all-in-one construct carrying GTR and Cas9, we disrupted the IGFBP3, MSTN, MC4R, and SOCS2 genes in multiple codon regions in one PFF cell simultaneously. This technique allows the simultaneous disruption of four genes with 5.5% efficiency. As a result, this approach may effectively target multiple genes at the same time, making it a powerful tool for establishing multiple genes mutant cells in pigs.</p

Table1_Distribution of Cambrian Source Rock Controlled by the Inherited Paleotopography on the Precambrian Basement in the Tarim Basin, NW China.DOCX

The Cambrian source rock is a key indicator that will guide future hydrocarbon explorations in ultra-deep strata. Despite its importance, the characteristics of paleotopography on the Precambrian basement and the distribution of source rocks above it remain unclear. The seismic and borehole data, isopach data, and the unconformity analysis are used to study the relations of the Cambrian source rock distribution to the paleotopography. The consistency of the strata thickness distribution maps of the Nanhua–Cambrian sequences show that the extensive thermal subsidence lasted the whole Sinian period, and makes the paleotopography of the Precambrian basement inherited from the landform after the Nanhua continental rifting. A major transgression caused by rapid subsidence of the Tarim Basin took place during the Cambrian. Shales were deposited offshore along the slopes of a paleo-uplift on the inherited paleotopography and formed the onlap in seismic profiles. The onlap on the inherited paleotopography reflects the distribution range of Yuertusi source rocks in the Cambrian.</p

Competitive Host–Guest Interaction between β‑Cyclodextrin Polymer and Pyrene-Labeled Probes for Fluorescence Analyses

We developed a novel homogeneous fluorescence analysis based on a novel competitive host–guest interaction (CHGI) mechanism between β-cyclodextrin polymer (polyβ CD) and pyrene-labeled probe for biochemical assay. Pyrene labeling with oligonucleotide strands can be recruited and reside in lipophilic cavities of polyβ CD. This altered lipophilic microenvironment provides favored polarity for enhanced quantum efficiencies and extraordinarily increases the luminescence intensity of pyrene. However, with addition of complementary DNA, the pyrene-labeled probe formed double-strand DNA to hinder pyrene from entering the cavities of polyβ CD. The release of pyrene from polyβ CD, which are followed by fluorescence extinguishing, will provide the clear signal turn-off in the presence of target DNA. We also introduced Exodeoxyribonuclease I (Exo I) and Exodeoxyribonuclease III (Exo III) to improve the sensitivity of this system, and the following product of cleavage reaction, pyrene-nucleotide, could more easily host–guest interact with polyβ CD and emit stronger fluorescence than pyrene-labeled probe. In addition, the successful detection of adenosine is also demonstrated by using the similar sensing scheme. Although this scheme might be easily interfered by some biomolecules in the real test sample, it holds promising potential for detecting a broad range of other types of aptamer-binding chemicals and biomolecules