Okara is a major agro-waste produced from the soybean industry. To hydrolyze the okara and enable nutrient release, a strategy to valorize okara using solid-state fermentation with food grade Bacillus subtilis ( B . subtilis ) WX-17 was carried out. The study showed that fermentation of okara with B . subtilis WX-17 improved its overall nutritional content. The total amino acids content increased from 3.04 ± 0.14 mg/g in unfermented okara to 5.41 ± 1.21 mg/g in okara fermented with B . subtilis WX-17. Total fatty acids content increased from 153.04 ± 5.10 to 166.78 ± 2.41 mg/g okara, after fermentation. Antioxidant content (DPPH) also increased by 6.4 times after fermentation. To gain insight into the mechanism, gas chromatography–mass spectrometry analysis was carried out. In total, 49 metabolites were detected, which could be classified mainly into carbohydrates, TCA cycle metabolites, amino acids and fatty acids. The decrease in carbohydrate metabolites, showed that glycolysis was upregulated. This would have provided the energy and metabolic flux towards the amino acid and fatty acid pathway. This is also in line with the increased amino acids and fatty acid production seen in okara fermented with B . subtilis WX-17. The findings of this work demonstrated the potential of using B. subtilis WX-17 fermentation, to enhance the nutritional profile of okara. This could serve as a potential low-cost animal feed or incorporated into the human diet.
Generation of induced pluripotent stem (iPS) cells from somatic cells has been achieved successfully by simultaneous viral transduction of defined reprogramming transcription factors (TFs). However, the process requires multiple viral vectors for gene delivery. As a result, generated iPS cells harbor numerous viral integration sites in their genomes. This can increase the probability of gene mutagenesis and genomic instability, and present significant barriers to both research and clinical application studies of iPS cells. In this paper, we present a simple lentivirus reprogramming system in which defined factors are fused in-frame into a single open reading frame (ORF) via self-cleaving 2A sequences. A GFP marker is placed downstream of the transgene to enable tracking of transgene expression. We demonstrate that this polycistronic expression system efficiently generates iPS cells. The generated iPS cells have normal karyotypes and are similar to mouse embryonic stem cells in morphology and gene expression. Moreover, they can differentiate into cell types of the three embryonic germ layers in both in vitro and in vivo assays. Remarkably, most of these iPS cells only harbor a single copy of viral vector. This system provides a valuable tool for generation of iPS cells, and our data suggest that the balance of expression of transduced reprogramming TFs in each cell is essential for the reprogramming process. More importantly, when delivered by non-integrating genedelivery systems, this re-engineered single ORF will facilitate efficient generation of human iPS cells free of genetic modifications.
Despite the advantages of using photodynamic therapy (PDT) for the treatment of head and neck tumors, it can only be used to treat early stage flat lesions due to the limited tissue penetration ability of the visible light. Here, we developed near-infrared (NIR) excitable upconversion nanoparticle (UCN) based PDT agent that can specifically target epithelial growth factor receptor (EGFR) overexpressing oral cancer cells, in a bid to widen the application of PDT against thick and solid advanced or recurrent head and neck cancers. In vivo studies using the synthesized anti-EGFR-PEG-TiO2-UCNs following systemic administration displayed no major sub-acute or long term toxic effects in terms of blood biochemical, hematological or histopathological changes at a concentration of 50 mg/kg. NIR-PDT even in the presence of a 10 mm tissue phantom placed over the xenograft tumor, showed significant delay in tumor growth and improved survival rate compared to conventional chlorin-e6 (Ce6) PDT using 665 nm red light. Our work, one of the longest study till date in terms of safety (120 d), PDT efficacy (35 d) and survival (60 d), demonstrates the usefulness of UCN based PDT technology for targeted treatment of thick and bulky head and neck tumors.
Brewers’ spent grains (BSG) are underutilized food waste materials produced in large quantities from the brewing industry. In this study, solid state fermentation of BSG using Bacillus subtilis WX-17 was carried out to improve the nutritional value of BSG. Fermenting BSG with the strain WX-17, isolated from commercial natto, significantly enhanced the nutritional content in BSG compared to unfermented BSG, as determined by the marked difference in the level of metabolites. In total, 35 metabolites showed significant difference, which could be categorized into amino acids, fatty acids, carbohydrates, and tricarboxylic acid cycle intermediates. Pathway analysis revealed that glycolysis was upregulated, as indicated by the drop in the level of carbohydrate compounds. This shifted the metabolic flux particularly towards the amino acid pathway, leading to a 2-fold increase in the total amount of amino acid from 0.859 ± 0.05 to 1.894 ± 0.1 mg per g of BSG after fermentation. Also, the total amount of unsaturated fatty acid increased by 1.7 times and the total antioxidant quantity remarkably increased by 5.8 times after fermentation. This study demonstrates that novel fermentation processes can value-add food by-products, and valorized food waste could potentially be used for food-related applications. In addition, the study revealed the metabolic changes and mechanisms behind the microbial solid state fermentation of BSG.
Pluronics F127 or (PEG)99–(PPG)69–(PEG)99 (with PEG and PPG representing polyethylene glycol and polypropylene glycol, respectively) was chemically modified by reacting with poly(polytetrahydrofuran carbonate) (PTHF) diol using a standard poly(urethane) reaction. The poly(F127/PTHF urethane)s showed lower critical gelation concentration as compared with Pluronics F127. The solution properties of these polymers were investigated at different temperatures using a hydrophobic dye probe. The thermodynamic variables associated with micelle formation were determined. This study uncovered valuable structure–property relationships affecting the micellization and gelation behavior. We tested these materials as potential ophthalmic delivery agents for the sustained release of natamycin for the treatment of eye infections. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2014, 131, 39924.
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