High-efficient separation of deoxyribonucleic acid from pathogenic bacteria by hedgehog-inspired magnetic nanoparticles microextraction.

Efficient separation of deoxyribonucleic acid (DNA) through magnetic nanoparticles (MN) is a widely used biotechnology. Hedgehog-inspired MNs (HMN) possess a high-surface-area due to the distinct burr-like structure of hedgehog, but there is no report about the usage of HMN for DNA extraction. Herein, to improve the selection of MN and illustrate the performance of HMN for DNA separation, HMN and silica-coated Fe3O4 nanoparticles (Fe3O4@SiO2) were fabricated and compared for the high-efficient separation of pathogenic bacteria of DNA. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are typical Gram-negative and Gram-positive bacteria and are selected as model pathogenic bacteria. To enhance the extraction efficiency of two kinds of MNs, various parameters, including pretreatment, lysis, binding and elution conditions, have been optimized in detail. In most separation experiments, the DNA yield of HMN was higher than that of Fe3O4@SiO2. Therefore, a HMN-based magnetic solid-phase microextraction (MSPE) and quantitative real-time PCR (qPCR) were integrated and used to detect pathogenic bacteria in real samples. Interestingly, the HMN-based MSPE combined qPCR strategy exhibited high sensitivity with a limit of detection of 2.0 × 101 CFU mL-1 for E. coli and 4.0 × 101 CFU mL-1 for S. aureus in orange juice, and 2.8 × 102 CFU mL-1 for E. coli and 1.1 × 102 CFU mL-1 for S. aureus in milk, respectively. The performance of the proposed strategy was significantly better than that of commercial kit. This work could prove that the novel HMN could be applicable for the efficient separation of DNA from complex biological samples.

Proteomics revealed the crosstalk between copper stress and cuproptosis, and explored the feasibility of curcumin as anticancer copper ionophore.

As an essential micronutrient element in organisms, copper controls a host of fundamental cellular functions. Recently, copper-dependent cell growth and proliferation have been defined as "cuproplasia". Conversely, "cuproptosis" represents copper-dependent cell death, in a nonapoptotic manner. So far, a series of copper ionophores have been developed to kill cancer cells. However, the biological response mechanism of copper uptake has not been systematically analyzed. Based on quantitative proteomics, we revealed the crosstalk between copper stress and cuproptosis in cancer cells, and also explored the feasibility of curcumin as anticancer copper ionophore. Copper stress not only couples with cuproptosis, but also leads to reactive oxygen species (ROS) stress, oxidative damage and cell cycle arrest. In cancer cells, a feedback cytoprotection mechanism involving cuproptosis mediators was discovered. During copper treatment, the activation of glutamine transporters and the loss of Fe-S cluster proteins are the facilitators and results of cuproptosis, respectively. Through copper depletion, glutathione (GSH) blocks the cuproptosis process, rescues the activation of glutamine transporters, and prevents the loss of Fe-S cluster proteins, except for protecting cancer cells from apoptosis, protein degradation and oxidative damage. In addition, the copper ionophore curcumin can control the metabolisms of lipids, RNA, NADH and NADPH in colorectal cancer cells, and also up-regulates positive cuproptosis mediators. This work not only established the crosstalk between copper stress and cuproptosis, but also discolored the suppression and acceleration of cuproptosis by GSH and curcumin, respectively. Our results are significant for understanding cuproptosis process and developing novel anticancer reagents based on cuproptosis.

Prediction of tannin profile in grape (Vitis vinifera L.) skins during berry maturation using a rapid mechanical puncture approach.

Tannin structure and composition are variable during grape maturation, and crucially determine perceived astringency, body structure and aging capacity of red wines. This study investigated the evolution of condensed tannins (CTs) in grape skins as maturation progressed and the feasibility of using a rapid mechanical puncture approach for assessing the CTs profile. The results showed that the mean degree of polymerization (mDP), molecular mass (MM), and proportions of (-)-epigallocatechin in extension subunits (EGC_ext) and (-)-epicatechin-3-O-gallate in terminal subunits (ECG_term) of skins increased during grape maturation, while CTs content and the proportion of (-)-epicatechin-3-O-gallate in extension subunits decreased. The predictive models built by random forest for CTs content based on skin weight, mDP, MM_subunit, EGC_ext, and ECG_term obtained good results with high squared correlation coefficients of prediction and calibration (R2_P > 0.85 and R2_C ≈ 0.95). In addition, the classifications of CTs characteristics obtained from ripe and unripe samples were observed in different principal component spaces. This study indicated that the mechanical properties were useful for predicting skin CTs profile, estimating tannin maturity stages, and providing information for optimal harvesting and winemaking protocols.

Effects of GATA6-AS/MMP9 on malignant progression of endometrial carcinoma.

PURPOSE: Previous studies have shown that long non-coding RNA (lncRNA) GATA6-AS is a tumor suppressor gene. However, the role of GATA6-AS in endometrial cancer (EC) has not been reported. We aimed at investigating the expression characteristics of GATA6-AS in EC tissues and cell lines, and explored whether it inhibits the malignant progression of EC through modulating matrix metalloproteinase-9 (MMP9). METHODS: GATA6-AS expression in 17 pairs of EC tissues and adjacent ones was studied by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Meanwhile, GATA6-AS expression levels in EC cell lines were also evaluated by qRT-PCR assay. In addition, GATA6-AS overexpression model was constructed using lentivirus in EC cell lines KLE and HEC-1B. The impacts of GATA6-AS overexpression model was constructed using lentivirus in EC cell lines KLE and HEC-1B on the proliferation capacity and apoptosis of EC cells were assessed by cell counting kit-8 (CCK-8), 5-Ethynyl-2'- deoxyuridine (EdU), and flow cytometry experiments. Furthermore, we explored the interaction between GATA6-AS and MMP9 in EC cells via performing luciferase assay and cell reverse experiments. RESULTS: Our data showed that GATA6-AS expression in EC tissue specimens was remarkably lower than that in adjacent ones. In vitro cell experiments revealed that overexpression of GATA6-AS markedly attenuated the proliferation ability of EC cells while elevated their apoptosis. Meanwhile, luciferase assay confirmed the binding relationship between GATA6-AS and MMP9. In addition, cell reverse experiments further demonstrated the mutual regulation between GATA6-AS and MMP9, which was, overexpression of MMP9 reversed the inhibitory influence of upregulation of GATA6-AS on the malignant progression of EC. CONCLUSIONS: lncRNA GATA6-AS, lowly expressed in EC tissue samples. Additionally, lncRNA GATA6-AS may suppress the malignant progression of EC through the modulation of regulating MMP9.

The changes of Lp-PLA2 in patients with gestational diabetes and its clinical significance.

ABSTRACT: Gestational diabetes mellitus (GDM) is one of the most common complications of pregnancy and associated with adverse pregnancy outcomes. The aim of this study was to investigate the changes of lipoprotein-associated phospholipaseA2 (Lp-PLA2) level and its correlation with biochemical indexes in patients with GDM.This observational cross-sectional study was performed among 52 GDM and 48 healthy pregnant women. Automatic biochemical analyzer was employed to test the biochemical indexes, including fasting plasma glucose (FPG), Hemoglobin A1c (HbA1c), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). The lipoprotein-associated phospholipaseA2 (Lp-PLA2) level was evaluated by enzyme-linked immunosorbent assay, and homeostatic model assessment for insulin resistance (HOMA-IR) was calculated.The levels of FPG, HbA1c, HOMA-IR, TG, TC and LDL-C were significantly increased while high-density lipoprotein cholesterol (HDL-C) level was significantly decreased in the GDM group when compared with those in the control group. Lp-PLA2 level in maternal blood in the GDM group was significantly higher than that in the control group (199.125 ±â€Š23.494 vs165.825 ±â€Š15.576 ng/mL, P < .05) and logistic regression analysis further confirmed the association of Lp-PLA2 levels with GDM. Furthermore, Lp-PLA2 positively correlated with HOMA-IR, TC, and LDL-C.Our results confirmed the association of Lp-PLA2 with GDM. This broadens our knowledge on the pathophysiology of GDM and provides insights into the development of new targets for the prevention and treatment of GDM.

Dynamic changes in anthocyanin biosynthesis regulation of Cabernet Sauvignon (Vitis vinifera L.) grown during the rainy season under rain-shelter cultivation.

The grapevine (Vitis vinifera L.) berry coloring mechanism in response to seasonal rain during grape ripening remains poorly understood. Therefore, anthocyanin biosynthesis regulation, dynamic changes in anthocyanin accumulation, biosynthetic enzyme activities, and related gene expression patterns were investigated in Cabernet Sauvignon grown under rain-shelter cultivation and open-field cultivation. Results showed that anthocyanin biosynthesis was strongly repressed during the rainy season. Environmental fluctuation from seasonal rain provoked metabolic responses in grapes, and there was a significantly greater accumulation of most of the anthocyanins, mainly the compositions of non-acylated and non-methylated, under rain-shelter cultivation; these findings indicate that rain-shelter cultivation may help improve tolerance to seasonal rain-induced stresses. Obvious resilience was observed in anthocyanins of open-field-cultivated grapes at harvest. Hierarchical cluster analysis indicated strong correlations between anthocyanin contents, CHI and DFR activities, and VvMYB5b transcriptional level. These findings provide novel insight into the crucial factors that directly modulate anthocyanin biosynthesis and consequently control grape coloration.

[Effect of arginine vasopressin on membrane potential of dorsal root ganglion neurons in rats].

The effect of arginine vasopressin (AVP) on membrane potential of neurons from dorsal root ganglion (DRG) was examined in the rat by means of intracellular recording technique. The results showed that (1) AVP induced hyperpolarization in the membrane of most DRG neurons. (2) The membrane conductance of the DRG neurons increased by 19.32% following application of AVP (p<0.05). (3) Perfusion with balance sodium solution (BSS) containing Cd(2+) (blocker of Ca(2+) channel) instead of Na+ failed to affect the AVP-induced membrane hyperpolarization of the DRG neurons (p> 0.05). After perfusion with BSS containing tetraethylammonium (TEA), however, the extent of AVP-induced hyperpolarization was reduced (p<0.05). (4) The AVP-induced hyperpolarization of the neurons was blocked by the antagonist of AVP V(1) receptors. The results demonstrate that AVP induces hyperpolarization of most DRG neurons, which might be caused by K(+) outflow mediated by AVP V(1) receptors in the membrane of the neurons.