Overexpression of DCF1 Inhibits Glioma through Destruction of Mitochondria and Activation of Apoptosis Pathway

Gliomas are the most common brain tumors affecting the central nervous system and are associated with a high mortality rate. DCF1 is a membrane protein that was previously found to play a role in neural stem cell differentiation. In the present study, we found that overexpression of dcf1 significantly inhibited cell proliferation, migration, and invasion and dramatically promoted apoptosis in the glioblastoma U251 cell line. DCF1 deletion mutations in the functional region showed that the complete structure of DCF1 was necessary for apoptosis. Furthermore, significantly lower tumorigenicity was observed in athymic nude mice by transplanting U251 cells overexpressing dcf1. To decode the apoptosis induced by dcf1, mitochondrial structure and membrane potential in glioma cells were investigated and the results indicated obvious mitochondrial swelling, destruction of cristae, and a significant decline in membrane potential. Mechanismly, caspase-3 signaling was activated. Finally, endogenous dcf1 silence in U251 cells was investigated. Results showed a highly methylation at -1339 and -1322 position at dcf1 promoter sequence, revealing the causal relationship between dcf1 gene and tumorigencicity. The present study identified a previously unknown cancer apoptosis mechanism involving dcf1 overexpression and provided a novel approach to potentially treat glioma patients

Antimikrobna rezistencija i svojstva virulencije bakterije Enterococcus faecium izolirane u goveda s kliničkim mastitisom iz pokrajine Ningxia, Kina

This study was conducted to determine the antimicrobial resistance and virulence traits of 32 Enterococcus faecium isolates from clinical bovine mastitis cases in Ningxia Province, China. In total, 32 E. faecium isolates were taken from subclinical bovine mastitis on the basis of morphological characterization and biochemical testing, and screened for antimicrobial susceptibility. The virulence genes of the isolates were studied using polymerase chain reaction (PCR). The disc diffusion assay revealed a high occurrence of resistance against tetracycline (78.1%) and erythromycin (68.8%) in the E. faecium isolates. However, all tested E. faecium were susceptible to linezolid and vancomycin. Moreover, all E. faecium isolates harbored the erythromycin-resistant genes ermA, ermB and ermC, as well as the tetracycline-resistant genes tetK, tetL and tetM. Furthermore, all E. faecium isolates carried more than 3 of the tested virulence genes. The presence of agg (100%), cpd (100%), efaA (100%), gelE (93.4%), and esp (75.0%) was found most frequently in all the tested isolates. These findings are useful for making appropriate antimicrobial choices and developing antivirulence therapies for subclinical bovine mastitis caused by E. faecium in Ningxia Province, China.Istraživanje je provedeno kako bi se odredila antimikrobina rezistencija i svojstva virulencije izolata bakterije Enterococcus faecium uzetih u goveda s kliničkim mastitisom. U ukupno 32 izolata goveda iz pokrajine Ningxia u Kini, procijenjena je antimikrobna osjetljivost na temelju morfološke karakterizacije i biokemijskih pretraga. Geni virulencije izolata istraženi su polimeraznom lančanom reakcijom (PCR). Disk-difuzijski test je u izolatu bakterije E. faecium pokazao visoku pojavnost rezistencije na tetraciklin (78,1 %) i eritromicin (68,8 %). Svi su pretraženi izolati bili osjetljivi na linezolid i vankomicin i imali gene rezisentne na eritromicin ermA, ermB i ermC, kao i na tetraciklin, tetK, tetL i tetM. Osim toga svi izolati E. faecium nosili su više od tri istraživana gena virulencije. Najčešći geni bili agg (100 %), cpd (100 %), efaA (100 %), gelE (93,4 %) i esp (75,0 %). Ovi rezultati mogu u pokrajini Ningxia u Kini pridonijeti pravilnom izboru antimikrobnog lijeka i razvoju uspješne terapije za supklinički goveđi mastitis uzrokovan bakterijom E. faecium

Antisense inhibition of ATM gene enhances the radiosensitivity of head and neck squamous cell carcinoma in mice

<p>Abstract</p> <p>Background</p> <p>Treatment failure after radiotherapy of head and neck squamous cell carcinoma (HNSCC) could be a significant problem. Our objective is to sensitize SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo </it>through inhibiting ATM expression using antisense oligodeoxynucleotides (AS-ODNs), and investigate the potential mechanism of radiosensitization.</p> <p>Methods</p> <p>We designed and synthesized AS-ODNs that target ATM mRNA to reduce the ATM expression. The influence on the expression of ATM mRNA and protein in SCCVII cells were analysed by real-time quantitative PCR and western blotting respectively. Clonogenic survival assay was performed to detect the survival ability of SCCVII cells after irradiation, while flow cytometry used to analyse the cell cycle and apoptosis. The volume of solid tumors generated with SCCVII cells was measured, and cell apoptosis was analysed by TUNEL assay after irradiation.</p> <p>Results</p> <p>The relative ATM mRNA and protein expression in SCCVII cells treated with ATM AS-ODNs were decreased to 25.7 ± 3.1% and 24.1 ± 2.8% of that in untreated cells respectively (<it>P </it>< 0.05). After irradiation, the survival fraction (SF) of cells treated with ATM AS-ODNs was lower than that of other groups at the same dose of radiation (<it>P </it>< 0.05), while the percentage of cells in G2/M phase decreased and apoptotic rate of cells increased(<it>P </it>< 0.05). The inhibition rate in SCCVII cells solid tumor exposed to X-ray alone was 23.2 ± 2.7%, while it was 56.1 ± 3.8% in the group which irradiated in combination with the treatment of ATM AS-ODNs (<it>P </it>< 0.05). The apoptotic index for the group irradiated in combination with ATM AS-ODNs injection was 19.6 ± 3.2, which was significantly higher than that of others (<it>P </it>< 0.05)</p> <p>Conclusion</p> <p>Inhibition of ATM expression sensitized SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo</it>. The potential mechanism should be the defective G2/M cell cycle checkpoint control and enhanced radiation-induced apoptosis.</p

比利时

peer reviewedQuinoa is an excellent source of nutritional and bioactive components. Protein is considered a key nutritional advantage of quinoa grain, and many studies have highlighted the nutritional and physicochemical properties of quinoa protein. In addition, quinoa protein is a good precursor of bioactive peptides. This review focused on the biological properties of quinoa protein hydrolysate and peptides, and gave a summary of the preparation and functional test of quinoa protein hydrolysate and peptides. A combination of milling fractionation and solvent extraction is recommended for the efficient production of quinoa protein. The biological functionalities of quinoa protein hydrolysate, including antidiabetic, antihypertensive, anti-inflammatory, antioxidant activities, and so on, have been extensively investigated based on in vitro studies and limited animal models. Additionally, bioinformatics analysis, including proteolysis simulation, virtual screening, and molecular docking, provides an alternative or assistive approach for exploring the potential bioactivity of quinoa protein and peptides. Nevertheless, further research is required for industrial production of bioactive quinoa peptides, verification of health benefits in humans, and mechanism interpretation of observed effects

Preparation, purification, and identification of novel antioxidant peptides derived from Gracilariopsis lemaneiformis protein hydrolysates

Gracilariopsis lemaneiformis (G. lemaneiformis) protein was hydrolyzed with alkaline protease to obtain antioxidant peptides. The enzymatic hydrolysis conditions were optimized through single-factor and orthogonal experiments. The results showed that the optimal process parameters were using 2% of alkaline protease, and substrate concentration of 1 g/100 mL and hydrolyzed 2 h at pH 8.0. Gel filtration chromatography and RP-HPLC were adopted for isolating and purifying the antioxidant peptides from the G. lemaneiformis protein hydrolysate (GLPH). Three novel antioxidant peptides were identified as LSPGEL (614.68 Da), VYFDR (698.76 Da), and PGPTY (533.57 Da) by nano-HPLC-MS/MS. The results of ABTS free radical scavenging rate demonstrated PGPTY exhibited the best antioxidant activity (IC50 = 0.24 mg/mL). Moreover, LSPGEL, VYFDR, and PGPTY were docked with Keap1, respectively. The molecular docking results suggested PGPTY had smaller docking energy and inhibition constants than the other two peptides. Finally, the cell viability assay evidenced the protective effect exerted by the antioxidant peptide on H2O2-induced oxidative damage. Above findings showed the potential of using antioxidant peptides from GLPH as antioxidants

Exposure to low-level metalaxyl impacts the cardiac development and function of zebrafish embryos.

Metalaxyl is an anilide pesticide that is widely used to control plant diseases caused by Peronosporales species. In order to study the toxic effects, zebrafish embryos were exposed to metalaxyl at nominal concentrations of 5, 50 and 500 ng/L for 72 hr, and the cardiac development and functioning of larvae were observed. The results showed that metalaxyl exposure resulted in increased rates of pericardial edema, heart hemorrhage and cardiac malformation. The distance between the sinus venosus and bulbus arteriosus, stroke volume, cardiac output and heart rate were significantly increased in larvae exposed to 50 and 500 ng/L metalaxyl compared to solvent control larvae. Significant upregulation in the transcription of tbx5, gata4 and myh6 was observed in the 50 and 500 ng/L treatments, and that of nkx2.5 and myl7 was observed in the 5, 50 and 500 ng/L groups. These disturbances may be related to cardiac developmental and functional defects in the larvae. The activity of Na+/K+-ATPase and Ca2+-ATPase was significantly increased in zebrafish embryos exposed to 500 ng/L metalaxyl, and the mRNA levels of genes related to ATPase (atp2a11, atp1b2b, and atp1a3b) (in the 50 and 500 ng/L groups) and calcium channels (cacna1ab) (in the 500 ng/L group) were significantly downregulated; these changes might be associated with heart arrhythmia and functional failure

Comprehensive Evaluation of Tea Cultivars Suitable for Matcha Production Using Multivariate Statistical Analysis

Matcha was prepared from 36 tea cultivars grown in the same tea garden according to the shading requirements for fresh leaves to be used for the production of matcha and its 11 quality indexes such as sensory quality, major physicochemical properties and chroma values were analyzed. In order to select tea cultivars suitable for the manufacturing of matcha, a comprehensive evaluation model of matcha quality was established by cluster analysis (CA), principal component analysis (PCA) and multiple linear regression analysis. The CA results showed that the 36 cultivars could be divided into three groups. Matcha from group I had the best quality with green color, fresh and mellow taste, and low phenol/ammonia ratio. Matcha from group II had high phenol/ammonia ratio and strong astringent taste. Matcha from Group III, consisting of etiolated and albino cultivars, had poor color and aroma quality. The PCA results showed that the cumulative contribution rate of the first five principal components was 88.152%. Comprehensive evaluation of matcha using the evaluation function constructed based on the first five principal components showed that the top 10 cultivars were Zhongcha 102, Taicha 12, Zhongcha 108, Fuding Dahao, Meizhan, Fuding Dabai, Fuyun 6, Zi Mudan, Maolv and Yingshuang. The model describing the relationship between sensory quality and physicochemical properties established by multiple linear regression analysis was as follows: y = 3.167|a*| + 46.850 (R2 = 0.710, P < 0.001). The scores of matcha cultivars evaluated by this model were highly consistent with the comprehensive evaluation results based on principal components, indicating that the a* value of dried tea could be used as a representative index to evaluate the quality of matcha. The results of this study can provide a reference for evaluating the suitability of tea cultivars for matcha manufacturing

Evaluation of Suitability for Green Tea Processing of Different Tea Cultivars Based on Multivariate Statistical Analysis

Principal component analysis (PCA) and multiple linear regression analysis (MLRA) were used to analyze and investigate the suitability of green tea for tea tree varieties in different tea cultivars. The fresh leaves of 63 tea cultivars planted in the same garden were processed into roasted green tea by the same method. The sensory and major physicochemical qualities (tea polyphenols, free amino acids, water extracts and chlorophyll) and the color (L*, a* and b*) of green tea were analyzed. To evaluate the suitability of the tested cultivars for green tea processing and its major influential factors, the data obtained were analyzed by PCA and MLRA. The results showed that the coefficient of variation (CV) of chlorophyll content in different cultivars was the highest (26.7%), followed by tea a* value (22.2%), sensory score for tea infusion color (17.1%) and sensory score for tea color (16.2%). A significant correlation was found between the polyphenol content of green tea and its color, tea infusion color and the color of infused tea leaves; between the chlorophyll content, |a*| and b* values of green tea and its color; between the chlorophyll content and |a*| value of green tea and the color of infused tea leaves; and between the |a*| and b* values of green tea and tea infusion color (P < 0.01). The PCA results showed that the contribution rate of the first five principal components (PCs) was 76.895%, and that of the first principal component was 31.918%, mainly pointing to the color quality. According to the evaluation model constructed based on the first five PCs, the top 10 cultivars were Zhongcha 108, Wuniuzao, Pingyangtezao, Mengshan 9, Soubeizhong, Fudingdabai, Jiukeng 16, Fudingdahao, Maolv, and Longjing 43. Using MLRA, the regression function between overall sensory score (Y) and |a*| value (x) was obtained as Y = 68.668 + 5.174x (R2 = 0.313) (P < 0.001). The top 10 varieties determined from this equation were highly consistent with the results of PCA, indicating that a* value is an important indicator for the evaluation of the suitability of tea cultivars for green tea processing