Knockdown of EIF2AK2-OAS1 axis reduces ATP production inducing AMPK phosphorylation to inhibit the malignant behavior of gastric cancer cells.

Energy metabolism has always been a hot topic in cancer progression and targeted therapy, and exploring the role of genes in energy metabolic pathways in cancer cells has become key to address this issue. Eukaryotic translation initiation factor 2α kinase 2 (EIF2AK2) plays regulatory roles in cancer and disorders of energy metabolism. Indeed, the role of EIF2AK2 in energy metabolism has been underestimated. The aim of this study is to reveal the expression specificity of EIF2AK2 in gastric cancer (GC) progression and metastasis, and to demonstrate the role of EIF2AK2 in energy metabolism, cytoskeleton, proliferation, death and metastasis pathways in GC cells. Mechanistically, EIF2AK2 overexpression promoted cytoskeleton remodeling and ATP production, mediated cell proliferation and metastasis, upregulated OAS1 expression, decreases p-AMPK expression and inhibited apoptosis in GC cells. Conversely, knockdown of EIF2AK2 resulted in the opposite effect. However, overexpression of OAS1 mediated the upregulation of mitochondrial membrane potential and promoted ATP production and NAD+/NADH ratio, but knockdown of OAS1 inhibited the above effects. In addition, knockdown of OAS1 had no effect on EIF2AK2 expression, but inhibited AMPK and upregulated p-AMPK expression. In conclusion, our study identified EIF2AK2 and OAS1 as previously undescribed regulators of energy metabolism in GC cells. We hypothesized that EIF2AK2-OAS1 axis may regulate energy metabolism and inhibit cellular malignant behavior in cancer cells by affecting ATP production to induce AMPK phosphorylation, suggesting EIF2AK2 as a potential therapeutic target for cancer cell progression.

Effects of protein concentration, ionic strength, and heat treatment on the interfacial and emulsifying properties of coconut (Cocos nucifera L.) globulins.

This research aimed to investigate the effects of protein concentration (0.2 %-1.0 %), ionic strength (100-500 mM NaCl), and heat treatment (temperature: 80 and 90℃; time: 15 and 30 min) on the interfacial and emulsifying properties of coconut globulins (CG). When protein concentration was set at 0.2-0.6 %, the interfacial adsorption increased with the increasing of protein concentration. However, the lowest interfacial viscoelasticity was found when CG concentration was 0.6 %. When the protein concentration was higher than 0.6 %, the dilatational viscoelasticity increased with the increasing of protein concentration. The protein concentration showed positive effect on the emulsion stability of CG. The ionic strength showed positive effect on the interfacial adsorption but negative effects on the interfacial viscoelasticity and emulsion stability. Higher temperature and longer heating time brought worse interface behavior. The heated CG (90℃, 30 min) had the worst interfacial behavior but the best emulsion stability.

Insight of the Functional and Biological Activities of Coconut (Cocos nucifera L.) Protein by Proteomics Analysis and Protein-Based Bioinformatics.

Coconut (Cocos nucifera L.) is one of the most critical economic crops in the tropics and sub-tropics. Although coconut protein has attracted more and more attention due to its nutritional potential, the lack of proteomic information has limited its practical application. The present study aimed to investigate the coconut meat proteome by shotgun proteomics and protein-based bioinformatic analysis. A grand total of 1686 proteins were identified by searching the National Center for Biotechnology Information (NCBI) protein database and self-constructed C. nucifera transcriptome repository. Among them, 17 and 9 proteins were identified as antioxidant proteins and globulins, respectively. Network analysis of the globulins referred to the sub-works of Cupin and Oleosin, and the antioxidant proteins were related to the sub-networks of glutathione metabolism and peroxisome. The bioactive peptides acquired by in-silico digestion of the targeted proteins have the potential to be applied as antioxidants and emulsifiers for both healthcare and food stabilization.

Effects of heat treatment and pH on the physicochemical and emulsifying properties of coconut (Cocos nucifera L.) globulins.

The present study aimed to assess the effects of heat treatment (70-90 °C) and pH (pH 3-11) on the physicochemical, structural, and emulsifying properties of coconut globulins (CG). The results revealed that the emulsifying property was improved with increasing temperature due to the denaturation degree of CG. CG had a better emulsifying property at pH 3 but showed the worst emulsifying property at pH 5 due to its lowest solubility, surface hydrophobicity, and absolute value of zeta potential. The best emulsifying stability was detected at pH 11 with 90 °C heating. SDS-PAGE indicated that the formation of aggregates cross-linked by covalent bonds was the main reason for the better emulsion stability at pH 3 and pH 11 with 90 °C heating. The secondary structure showed that CG had more α-helix and ß-turn contents as well as fewer ß-sheet contents at pH 3 and pH 11 with 90 °C heating.

Single-tube analysis for ultra-fast and visual detection of Salmonella.

Herein, we developed an ultra-fast and visual single-tube nucleic acid detection approach, which combined the advantages of self-settling characteristics of chitosan-functionalized diatomaceous earth (CDE) and accelerated PCR (AC-PCR). DNA was rapidly extracted by CDE within 3 min for the next nucleic acid amplification based on the nucleic acid attached on the chitosan in pH = 5.0. Under the action of gravity, the DNA-enriched CDE self-sediments to the bottom of the tube could be directly used for AC-PCR to achieve single-tube extraction and amplification. Our method detected Salmonella culture fluids with a detection limit of 1 CFU/mL, which was 100-fold more sensitive than conventional method that have not undergone nucleic acid enrichment. Furthermore, it also displayed high specificity and sensitivity for a variety of spiked samples. The entire process could be completed within 17 min in a single tube, and in particular, the result was visualized by the naked eyes. Overall, it is an all-in-one detection strategy without the requirement of redundant procedure, which greatly improved the detection efficiency, and saved the time and the cost. With these advantages, the approach will supply a promising tool in the field of point-of-care testing for Salmonella and other foodborne pathogens.

Expression of p53ß and Δ133p53 isoforms in different gastric tissues.

This study aims to detect the mRNA of p53ß and Δ133p53 isoforms in three gastric carcinoma cell lines and tissues of superficial gastritis, atrophic gastritis, gastric carcinoma, or paracancerous area. Nested reverse transcription PCR was used to detect the mRNA of p53ß and Δ133p53 isoforms in tissues of superficial gastritis, chronic atrophic gastritis, gastric cancer cell lines (SGC-7901, MKN45, KATO III), gastric adenocarcinoma, and paracancerous lesion. The amplified products were shown by agarose gel electrophoresis. The expression difference among various tissues was analyzed by x(2) tests. The positive rates of ∆133p53 mRNA were 73.3% (11/15) in gastric adenocarcinoma and 20% (3/15) in paracancerous tissue, whereas the positive rates of p53ß mRNA were 20% (3/15) in gastric adenocarcinoma and 66.7% (10/15) in paracancerous tissue. The difference between adenocarcinoma and paracancerous tissues was significant (P<0.05). The positive rates of ∆133p53 mRNA were 25% (5/20), 50% (15/30), and 75% (15/20), respectively, in superficial gastritis, atrophic gastritis, and gastric adenocarcinoma; the positive rates of p53ß mRNA were 65% (13/20), 33.3% (10/30), and 25% (5/20), respectively, in superficial gastritis, atrophic gastritis, and gastric adenocarcinoma. The difference between adenocarcinoma and superficial gastritis samples was significant (P<0.05). Both p53ß and ∆133p53 mRNAs were positive in MKN45; only p53ß mRNA was detected in SGC7901; neither p53ß nor ∆133p53 mRNA was detected in KATO III. ∆133p53 and p53ß, which are possible indicators for the diagnosis and biological therapy of gastric carcinoma, were expressed differentially in different gastric tissues.

Role of p53ß in the inhibition of proliferation of gastric cancer cells expressing wild-type or mutated p53.

p53 is a tumor suppressor gene whose mutation is highly associated with tumorigenesis. The present study investigated the role of p53ß in the inhibition of proliferation of gastric cancer cell lines expressing wild-type or mutated p53. Wild-type p53 is expressed in MKN45 cells, but deleted in KATOIII cells, whereas mutated p53 is expressed in SGC7901 cells. The mRNA expression levels of p53ß and Δ133p53 were detected in MKN45, SGC-7901 and KATOIII gastric cancer cell lines using nested polymerase chain reaction (PCR). The mRNA expression levels of p53, p53ß and B-cell lymphoma 2-associated X protein (Bax) were detected in the MKN45 and SGC-7901 cells following treatment with cisplatin by reverse transcription-PCR. The inhibition of cellular proliferation following treatment with cisplatin was measured by MTT assay. The results of the present study demonstrated that both p53ß and Δ133p53 mRNA were expressed in the MKN45 cells, whereas only p53ß mRNA was expressed in the SGC7901 cells. No expression of p53ß or Δ133p53 mRNA was detected in the KATOIII cells. Following treatment with cisplatin, the number of both MKN45 and SGC-7901 cells was significantly reduced (P<0.001). In the MKN45 cells, p53ß, p53 and Bax mRNA expression levels gradually increased with the dose of cisplatin, and the expression of p53ß was positively correlated with the expression of p53 (tr=6.358, P<0.05) and Bax (tr=8.023, P<0.05). In the SGC-7901 cells, the expression levels of p53ß, p53 and Bax mRNA did not alter with the dose of cisplatin, and the expression of p53ß was positively correlated to the expression of p53 (tr=26.41, P<0.01) but not that of Bax. The present study identified the different roles of the p53ß isoform in gastric cancer cells with different p53 backgrounds. Enhanced knowledge regarding the p53 status is required for the development of specific biological therapies against gastric cancer.