Electrochemical Oxidation of 2’-deoxyguanosine-5’-triphosphate on Ionic Liquid Modified Carbon Paste Microelectrode and its Sensitive Detection

Electrochemical oxidation of 2’-deoxyguanosine-5’-triphosphate (dGTP) was investigated on the ionic liquid modified carbon paste microelectrode by cyclic voltammetry in this paper. The carbon io-nic liquid microelectrode (mCILE) was prepared with ionic liquid 1-ethyl-3-methylimidazolium ethylsul-fate ([Emim][EtOSO3]) as the modifier. Then the electrochemical process of dGTP on the mCILE was recorded with the electrochemical parameters calculated and the oxidation mechanism discussed. Under the optimal conditions differential pulse voltammetric peak current was proportional to the dGTP concen-tration in the range from 1.0×10–6 M to 7.0×10–4 M with the linear regression equation as log(Ipa/A) = 0.77 log(c/mol dm–3) – 4.082 (γ = 0.999). The proposed method showed good selectivity and stability without the interferences of coexisting substances. (doi: 10.5562/cca1737

Isotopic dynamics of precipitation and its regional and local drivers in a plateau inland lake basin, Southwest China

Shrinkage of plateau lakes under climate strength has drawn growing attention. Because of its intricate implication to hydro-meteorological condition and climate system, stable isotopes in precipitation (e.g. delta H-2(p) and delta O-18(p)) provide us a powerful tool to understand the climate-hydrologic dynamics in shrinking lakes. However, how the regional atmospheric circulation, moisture sources and local fractionation processes drive isotopic variability from temporal to spatial scale has rarely been reported for remote plateau lakes. Hence, we collected a total of 98 rainfall samples at the south and the north shores of Chenghai lake, Yunnan-Guizhou Plateau to study the potential driving forces of precipitation isotope variability during the wet season of 2019. Based on backward trajectories of air masses obtained from HYSPLIT model, 68% of moisture came from delta O-18 depleted ocean (Indian Ocean, Bay of Bengal, South China Sea and Pacific Ocean), and the rainout process promoted the isotopic depletion when moisture arrived at the study basin. Evapotranspiration increased the heavy isotope ratios in precipitation originated from continents (northern China inland and western continents). The temporal dynamics of delta O-18(p) and delta H-2(p) were in phase with the convection activities intensity underlined the influence from large-scale atmospheric circulation. Local meteorological factors played a secondary role in isotope variability. Precipitation amount-effect strongly affected isotope ratios while mild anti-temperature effect was observed at daily scale. Interestingly, the rainfall isotope ratios showed different mechanisms in govern at lake south shore and north shore, with a distance of 19 km in between. This south-to-north difference can be explained by either lower 1.03% sub-evaporation in the south shore or 7% of recycled moisture contributing to precipitation in the north shore. Our findings discover the driving forces for delta O-18(p) variation and provide solid interpretations for hydro-climate change in Southwest China. (C) 2020 Elsevier B.V. All rights reserved.Peer reviewe

PAHs in the North Atlantic Ocean and the Arctic Ocean: Spatial Distribution and Water Mass Transport

In the Arctic Ocean, it is still unclear what role oceanic transport plays in the fate of semivolatile organic compounds. The strong-stratified Arctic Ocean undergoes complex inputs and outputs of polycyclic aromatic hydrocarbons (PAHs) from the neighboring oceans and continents. To better understand PAHs’ transport processes and their contribution to high-latitude oceans, surface seawater, and water column, samples were collected from the North Atlantic Ocean and the Arctic Ocean in 2012. The spatial distribution of dissolved PAHs (∑9PAH) in surface seawater showed an “Arctic Shelf \u3e Atlantic Ocean \u3e Arctic Basin” pattern, with a range of 0.3–10.2 ng L−1. Positive matrix factorization modeling results suggested that vehicle emissions and biomass combustion were the major PAHs sources in the surface seawater. According to principal component analysis, PAHs in different water masses showed unique profiles indicating their different origins. Carried by the Norwegian Atlantic Current (0–800 m) and East Greenland Current (0–300 m), PAH individuals’ net transport mass fluxes ranged from −4.4 ± 1.7 to 53 ± 39 tons year−1 to the Arctic Ocean. We suggested the limited contribution of ocean currents on PAHs’ delivery to the Arctic Ocean, but their role in modulating PAHs’ air–sea interactions and other biogeochemical processes needs further studies

Tissue factor pathway inhibitor-2 was repressed by CpG hypermethylation through inhibition of KLF6 binding in highly invasive breast cancer cells

<p>Abstract</p> <p>Background</p> <p>Tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated Kunitz inhibitor that inhibits plasmin and trypsin-mediated activation of zymogen matrix metalloproteinases involved in tumor progression, invasion and metastasis. Here, we have investigated the mechanism of DNA methylation on the repression of TFPI-2 in breast cancer cell lines.</p> <p>Results</p> <p>We found that both protein and mRNA of TFPI-2 could not be detected in highly invasive breast cancer cell line MDA-MB-435. To further investigate the mechanism of TFPI-2 repression in breast cancer cells, 1.5 Kb TFPI-2 promoter was cloned, and several genetic variations were detected, but the promoter luciferase activities were not affected by the point mutation in the promoter region and the phenomena was further supported by deleted mutation. Scan mutation and informatics analysis identified a potential KLF6 binding site in TFPI-2 promoter. It was revealed, by bisulfite modified sequence, that the CpG island in TFPI-2 promoter region was hypermethylated in MDA-MB-435. Finally, using EMSA and ChIP assay, we demonstrated that the CpG methylation in the binding site of KLF-6 diminished the binding of KLF6 to TFPI-2 promoter.</p> <p>Conclusion</p> <p>In this study, we found that the CpG islands in TFPI-2 promoter was hypermethylated in highly invasive breast cancer cell line, and DNA methylation in the entire promoter region caused TFPI-2 repression by inducing inactive chromatin structure and decreasing KLF6 binding to its DNA binding sequence.</p

A comparison of growth on mercuric chloride for three Lemnaceae species reveals differences in growth dynamics that effect their suitability for use in either monitoring or remediating ecosystems contaminated with mercury

Mercury (Hg) is a toxic heavy metal that can alter the ecological balance when it contaminates aquatic ecosystems. Previously, researchers have used various Lemnaceae species either to monitor and/or remove heavy metals from freshwater systems. As Hg contamination is a pressing issue for aquatic systems worldwide, we assessed its impact on the growth of three commonly species of Lemnaceae - Lemna gibba 6745, Lemna minor 6580 and Spirodela polyrhiza 5543. We exposed plants to different concentrations of mercuric chloride (HgCl₂) and monitored their growth, including relative growth rate, frond number (FN), and fresh weight (FW). These data were coupled with measurements of starch content, levels of photosynthetic pigment and the activities of antioxidant substances. The growth of all three lines showed significant negative correlations with Hg concentrations, and starch content, photosynthetic pigment, soluble protein and antioxidant enzymes levels were all clearly affected. Our results indicate that the L. gibba line used in this study was the most suitable of the three for biomonitoring of water contaminated with Hg. Accumulation of Hg was highest in the S. polyrhiza line with a bioconcentration factor over 1,000, making this line the most suitable of the three tested for use in an Hg bioremediation system

Expression of a LINE-1 endonuclease variant in gastric cancer: its association with clinicopathological parameters

BACKGROUND: Long interspersed nuclear element-1 (LINE-1 or L1), the most abundant and only autonomously active family of non-LTR retrotransposons in the human genome, expressed not only in the germ lines but also in somatic tissues. It contributes to genetic instability, aging, and age-related diseases, such as cancer. Our previous study identified in human gastric adenocarcinoma an upregulated transcript GCRG213, which shared 88% homology with human L1 sequence and contained a putative conserved apurinic/apyrimidinic endonucleas1 domain. METHODS: Immunohistochemistry was carried out by using a monoclonal mouse anti-human GCRG213 protein (GCRG213p) antibody produced in our laboratory, on tissue microarray constructed with specimens from 175 gastric adenocarcinoma patients. The correlation between GCRG213p expression and patient clinicopathological parameters was evaluated. GCRG213p expression in gastric cancer cell lines were studied using Western blotting analysis. L1 promoter methylation status of gastric cancer cells was tested using methylation-specific PCR. BLASTP was used at the NCBI Blast server to identify GCRG213p sequence to any alignments in the Protein Data Bank databases. RESULTS: Most primary gastric cancer, lymph node metastases and gastric intestinal metaplasia glands showed positive GCRG213p immunoreactivity. High GCRG213p immunostaining score in the primary gastric cancer was positively correlated with tumor differentiation (well differentiated, p = 0.001), Lauren’s classification (intestinal type, p < 0.05) and a late age onset of gastric adenocarcinoma (≥65 yrs; p < 0.05). GCRG213p expression has no association with other clinicopathological parameters, including survival. Western blotting analysis of GCRG213p expression in gastric cancer cells indicated that GCRG213p level was higher in gastric cancer cell lines than in human normal gastric epithelium immortalized cell line GES-1. Partial methylation of L1 in gastric cancer cells was confirmed by methylation-specific PCR. BLASTP program analysis revealed that GCRG213p peptide shared 83.0% alignment with the C-terminal region of L1 endonuclease (L1-EN). GCRG213p sequence possesses the important residues that compose the conserved features of L1-EN. CONCLUSIONS: GCRG213p could be a variant of L1-EN, a functional member of L1-EN family. Overexpression of GCRG213p is common in both primary gastric cancer and lymph node metastasis. These findings provide evidence of somatic L1 expression in gastric cancer, and its potential consequences in the form of tumor

Expression of GCRG213p, LINE-1 endonuclease variant, significantly different in gastric complete and incomplete intestinal metaplasia.

BACKGROUND: Intestinal metaplasia (IM) of the gastric mucosa is classified as complete (Type I) and incomplete IM (Type II and III) subtypes, which showed significantly different risk for developing to gastric adenocarcinoma (GAC). GCRG213, a variant of L1-endonuclease (L1-EN), first identified in our lab, was upregulated in GAC tissue. However, the relationship between GCRG213 and IM subtypes is not clear. Our study explored the association of GCRG213 protein (GCRG213p) with IM subtypes. METHODS: Gastric cancer and/or para-tumor tissue samples were collected from 123 patients who underwent gastrectomy for intestinal type gastric adenocarcinoma. The subtypes of IM were characterized with Alcian blue-periodic acid-Schiff and High Iron Diamine-Alcian blue staining methods. Immunohistochemistry of GCRG213p was performed, and its expression in gastric adenocarcinoma and para-tumor tissue including dysplasia, IM, and normal mucosa were analyzed. RESULTS: GCRG213p was expressed in 48.94% IM, 57.14% dysplasia and 55.32% GAC, respectively. GCRG213p expression was higher in well and moderately differentiated adenocarcinoma (P = 0.037). In IM glands, GCRG213p expressed mainly in the cytoplasm of absorptive enterocytes with defined brush borders, but not in goblet cells. The expression of GCRG213p in type I IM (90.00%) was significantly higher than that in type II (36.36%) and type III (25.00%) (P \u3c 0.001). In normal gastric mucosa, GCRG213p was exclusively positive in the cytoplasm of gastric chief cells. CONCLUSIONS: The expression of GCRG213p in complete IM was significantly higher than in incomplete IM, which implies that GCRG213p may play a role on the developing of IM to adenocarcinoma. GCRG213p was exclusively expressed in chief cells, suggesting that it might be involved in cell differentiation from the chief cells to IM