Isolation and extraction of glansreginin A from walnut meal and its effect on the proliferation of 3T3-L1 cells

Abstract Glansreginin A is an indicative component in walnut and is abundant in walnut meal. The aim of this study was to isolate and purify glansreginin A from the walnut meal, and to investigate the weight loss and lipid-lowering potential of glansreginin A by studying the effect of glansregin A on the proliferation of 3T3-L1 preadipocytes. Firstly, the response surface methodology was used to effectively improve the extraction yield of glansreginin A. The maximum extraction rate of glansreginin A was 0.363%, and the optimal extraction process conditions were determined. In addition, the lipid-lowering activity of glansreginin A was investigated by cell experiments. The results showed that glansreginin A could inhibit the proliferation of 3T3-L1 preadipocytes in a dose-dependent manner. And cell cycle of different groups of cells treated with glansreginin A was also measured using flow cytometry. The results showed most of the cells were blocked in G0/G1 phase and significantly decreased in S phase. These results suggest that glansreginin A could inhibit the proliferation of 3T3-L1 preadipocytes by causing cell cycle arrest. These findings provided a theoretical basis for the future research of glansreginin A and the development of slimming and fat-reducing foods

GRIM-19 Disrupts E6/E6AP Complex to Rescue p53 and Induce Apoptosis in Cervical Cancers

BACKGROUND: Our previous studies showed a down-regulation of GRIM-19 in primary human cervical cancers, and restoration of GRIM-19 induced tumor regression. The induction of tumor suppressor protein p53 ubiquitination and degradation by E6 oncoportein of high risk-HPV through forming a stable complex with E6AP is considered as a critical mechanism for cervical tumor development. The aims of this study were to determine the potential role of GRIM-19 in rescuing p53 protein and inducing cervical cancer cell apoptosis. METHODOLOGY/PRINCIPAL FINDINGS: The protein levels of GRIM-19 and p53 were detected in normal cervical tissues from 45 patients who underwent hysterectomy for reasons other than neoplasias of either the cervix or endometrium, and cervical cancer tissues from 60 patients with non-metastatic squamous epithelial carcinomas. Coimmunoprecipitation and GST pull-down assay were performed to examine the interaction of GRIM-19 with 18E6 and E6AP in vivo and in vitro respectively. The competition of 18E6 with E6AP in binding GRIM-19 by performing competition pull-down assays was designed to examine the disruption of E6/E6AP complex by GRIM-19. The augment of E6AP ubiquitination by GRIM-19 was detected in vivo and in vitro ubiquitination assay. The effects of GRIM-19-dependent p53 accumulation on cell proliferation, cell cycle, apoptosis were explored by MTT, flow cytometry and transmission electron microscopy respectively. The tumor suppression was detected by xenograft mouse model. CONCLUSION/SIGNIFICANCE: The levels of GRIM-19 and p53 were concurrently down regulated in cervical cancers. The restoration of GRIM-19 can induce ubiquitination and degradation of E6AP, and disrupt the E6/E6AP complex through the interaction of N-terminus of GRIM-19 with both E6 and E6AP, which protected p53 from degradation and promoted cell apoptosis. Tumor xenograft studies also revealed the suppression of p53 degradation in presence of GRIM-19. These data suggest that GRIM-19 can block E6/E6AP complex; and synergistically suppress cervical tumor growth with p53

Development Strategy of Microbial Industry in China

The microbial industry is a strategic emerging industry and it is an important field for developing new growth drivers and for creating new global competitive advantages. In the face of the great opportunity to promote the construction of Healthy China, the development of the microbial industry is significant for regional scientific and technological innovation, high-quality industrial development, and improvement of people’s welfare in China. The study expounded the macro-demand for the development of microbial industry and analyzed the development status of the global microbial industry in terms of industrial layout and technological progress. The study further outlined the development trend of the microbial industry in China from the aspects of industrial policy environment and industrial scale, investigated the development status of industrial segmentation directions including microbial safety, microbial health, microbial manufacturing, and microbial medicine, and dissected the challenges in the development of the industry. Our findings indicated that structuring systematic big science facilities, developing a series of industrialized technologies, and promoting the industrial application of microbial resources are key initiatives to accelerate the development of the microbial industry in China. We proposed to strengthen the research and development of key core technologies of the microbial high-tech industry, promote the establishment of an industrial policy system for collaborative innovation and development, and build industrial clusters centering around microbial innovation platforms, all of which will greatly improve the development level of microbial industry in China

Research Progress in Molecular Identification Technology for Microbial Resources

As the most widely distributed and abundant biological resources in the world, microbial resources have been applied in many fields and shown great economic and social values. Although there have been numerous previous studies on microorganisms, the studies could not be further launched owing to limited technical methods. However, the continuous development of microbial detection technologies now provides new approaches for microbial research. In this article, we summarize the molecular identification technologies and their application for microbial resources in China and abroad; these technologies include deoxyribonucleic acid (DNA) (G+C) mol% identification, nucleic acid hybridization, DNA fingerprinting, nucleic acid amplification, gene chips, and high throughput sequencing. Moreover, we summarize the advantages and disadvantages of each technology and analyze the problems faced by molecular identification of microbial resources, including reliance on laboratory environment, insufficient reproducibility and accuracy of experimental results, and lack of microbial databases. To protect and utilize microbial resources, China should strengthen the research of each identification technology, establish a microbial resource identification platform with independent intellectual property rights, and improve the microbial resource database

Antibacterial Regularity Mining Beneath the Systematic Activity Database of Lipopeptides Brevilaterins: An Instructive Activity Handbook for Its Food Application

Bacterial contamination is a primary threat to food safety. Therefore, the persistent development of natural antibacterial agents has become essential work. The present essay attempts to establish a systematic antibacterial activity database to instruct the food application of brevilaterins, promising antibacterial lipopeptides from Brevibacillus laterosporus S62-9. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were systematically collected from 43 species of standard bacteria and 140 strains of isolated bacteria (food spoilage bacteria and antibiotic-resistant bacteria) using a broth dilution method. The results showed that brevilaterins performed a broad-spectrum inhibitory (0.5~128 μg/mL) and bactericidal activity (1~256 μg/mL), especially efficient against Gram-positive bacteria and spoilage bacteria from grain products. Moreover, brevilaterins not only inhibit and kill multiple antibiotic-resistant bacteria but do not readily develop resistance, with a small specific value of MBC/MIC (1~8). Furthermore, brevilaterins would interact with negatively charged sodium dodecyl sulfate and bind amphipathic soybean phospholipid with an affinity constant of KD = 4.70 × 10−4 M. No significant activity difference was found between brevilaterin B and brevilaterin C. Collectively, this work contributed rich antibacterial data of brevilaterins and revealed the antibacterial regularity beneath these data, which can be used as an activity handbook to instruct their application in food safety

Selection, Identification, and Application of Aptamers against Agaricus bisporus Lectin to Establish an Aptamer-AuNPs Colorimetric Method for Detection of ABL

Agaricus bisporus lectin (ABL), which is one of the antinutritional factors in A. bisporus, is an important allergen and harmful to human health. Due to the shortcomings of the current detection methods, it is extremely urgent to establish a rapid and sensitive detection method for ABL in foods. To isolate the ssDNA aptamer of ABL, 13 rounds of subtractive systematic evolution of ligands by exponential enrichment (SELEX) selection were carried out. As a result, six candidate aptamers were selected and further examined for their binding affinity and specificity by enzyme-linked aptamer method. One aptamer (seq-41) against ABL with a high affinity and specificity was isolated and demonstrated to be the optimal aptamer whose dissociation constant reaches the nanomolar level, Kd = 31.17 ± 0.1070 nM. Based on seq-41, an aptamer-AuNPs colorimetric method was established to detect ABL with a linear range of 0.08∼1.70 μg/mL and the detection limit is 0.062 μg/mL. This study provides a novel aptamer-AuNPs colorimetric method with high sensitivity and specificity for detection of ABL and a novel strategy for development of detection method of fungal or plant allergens

Dietary Effect of <i>Brevibacillus laterosporus</i> S62-9 on Chicken Meat Quality, Amino Acid Profile, and Volatile Compounds

Probiotics are being used in diets to improve the quality of chicken meat. The aim of the study was to investigate the effects of dietary supplementation with Brevibacillus laterosporus S62-9 microbial agent on the meat quality, amino acids, and volatile compounds of chicken. The experiment was carried out with 160 1-day-old Arbor Acres male broiler chickens, rearing for 42 d. The chickens were randomly divided into two groups of 8 replicates each, with 10 chickens in each group. No supplement was added to the basal diet in the control group and Brevibacillus laterosporus S62-9 microbial agent was added to the diet of the experimental group. At the end of the experiment, the meat quality, meat chemical composition, amino acid composition, and volatile compounds of chicken were determined. The results showed that pH (p p p p p p Brevibacillus laterosporus S62-9 microbial agent can be used as a novel and effective feed supplement to improve the nutritional quality and flavor characteristics of broilers