How chip size impacts steam pretreatment effectiveness for biological conversion of poplar wood into fermentable sugars

Background Woody biomass is highly recalcitrant to enzymatic sugar release and often requires significant size reduction and severe pretreatments to achieve economically viable sugar yields in biological production of sustainable fuels and chemicals. However, because mechanical size reduction of woody biomass can consume significant amounts of energy, it is desirable to minimize size reduction and instead pretreat larger wood chips prior to biological conversion. To date, however, most laboratory research has been performed on materials that are significantly smaller than applicable in a commercial setting. As a result, there is a limited understanding of the effects that larger biomass particle size has on the effectiveness of steam explosion pretreatment and subsequent enzymatic hydrolysis of wood chips. Results To address these concerns, novel downscaled analysis and high throughput pretreatment and hydrolysis (HTPH) were applied to examine whether differences exist in the composition and digestibility within a single pretreated wood chip due to heterogeneous pretreatment across its thickness. Heat transfer modeling, Simons’ stain testing, magnetic resonance imaging (MRI), and scanning electron microscopy (SEM) were applied to probe the effects of pretreatment within and between pretreated wood samples to shed light on potential causes of variation, pointing to enzyme accessibility (i.e., pore size) distribution being a key factor dictating enzyme digestibility in these samples. Application of these techniques demonstrated that the effectiveness of pretreatment of Populus tremuloides can vary substantially over the chip thickness at short pretreatment times, resulting in spatial digestibility effects and overall lower sugar yields in subsequent enzymatic hydrolysis. Conclusions These results indicate that rapid decompression pretreatments (e.g., steam explosion) that specifically alter accessibility at lower temperature conditions are well suited for larger wood chips due to the non-uniformity in temperature and digestibility profiles that can result from high temperature and short pretreatment times. Furthermore, this study also demonstrated that wood chips were hydrated primarily through the natural pore structure during pretreatment, suggesting that preserving the natural grain and transport systems in wood during storage and chipping processes could likely promote pretreatment efficacy and uniformity

A novel HCC prognosis predictor PDSS1 affects the cell cycle through the STAT3 signaling pathway in HCC

Decaprenyl diphosphate synthase subunit 1 (PDSS1) is closely related to a variety of human diseases, but its expression pattern and biological function in HCC have not been studied to date.MethodsThe expression level of PDSS1 was analyzed using the TCGA and GEO databases. The relationships between PDSS1 and patient clinicopathological characteristics were verified based on TCGA clinical data. Additionally, the co-expressed genes of PDSS1were investigated and Gene Set Enrichment Analysis (GSEA) was conducted using LinkedOmics. Next, the association between PDSS1 and immune infiltration was determined using version 1.34.0 of the GSVA package. EdU assay, colony-formation assay, transwell assay, wound-healing assay, and flow cytometry analysis were used to assess the effect of PDSS1 on the cell phenotype.ResultsPDSS1 was upregulated in HCC compared with adjacent tissues. High PDSS1 in HCC was associated with poor overall survival, disease-specific survival, and progress-free interval. Results suggested that PDSS1 may activate multiple oncogenic pathways in HCC, especially those involved in the cell cycle. The expression of PDSS1 was significantly related to Th2 cells, TFH, T helper cells, NK CD56bright cells, cytotoxic cells, DC, CD8 T cells, and neutrophils. PDSS1 knockdown inhibited cell proliferation, cell cycle, migration and invasion. Furthermore, PDSS1 acted as an oncogene through the STAT3 signaling pathway.ConclusionOur study reveals that a high level of PDSS1 is significantly correlated with poor patient prognosis and immune cell infiltration in HCC. PDSS1 may be a novel biomarker and potential therapeutic target for HCC

Nutlin-3 overcomes arsenic trioxide resistance and tumor metastasis mediated by mutant p53 in Hepatocellular Carcinoma

Background: Arsenic trioxide has been demonstrated as an effective anti-cancer drug against leukemia and solid tumors both in vitro and in vivo. However, recent phase II trials demonstrated that single agent arsenic trioxide was poorly effective against hepatocellular carcinoma (HCC), which might be due to drug resistance. Methods: Mutation detection of p53 gene in arsenic trioxide resistant HCC cell lines was performed. The therapeutic effects of arsenic trioxide and Nutlin-3 on HCC were evaluated both in vitro and in vivo. A series of experiments including MTT, apoptosis assays, co-Immunoprecipitation, siRNA transfection, lentiviral infection, cell migration, invasion, and epithelial-mesenchy-mal transition (EMT) assays were performed to investigate the underlying mechanisms. Results: The acquisition of p53 mutation contributed to arsenic trioxide resistance and enhanced metastatic potential of HCC cells. Mutant p53 (Mutp53) silence could re-sensitize HCC resistant cells to arsenic trioxide and inhibit the metastatic activities, while mutp53 overexpression showed the opposite effects. Neither arsenic trioxide nor Nutlin-3 could exhibit obvious effects against arsenic trioxide resistant HCC cells, while combination of them showed significant effects. Nutlin-3 can not only increase the intracellular arsenicals through inhibition of p-gp but also promote the p73 activation and mutp53 degradation mediated by arsenic trioxide. In vivo experiments indicated that Nutlin-3 can potentiate the antitumor activities of arsenic trioxide in an orthotopic hepatic tumor model and inhibit the metastasis to lung. Conclusions: Acquisitions of p53 mutations contributed to the resistance of HCC to arsenic trioxide. Nutlin-3 could overcome arsenic trioxide resistance and inhibit tumor metastasis through p73 activation and promoting mutant p53 degradation mediated by arsenic trioxide

Diphenyl Difluoroketone: A Potent Chemotherapy Candidate for Human Hepatocellular Carcinoma

Diphenyl difluoroketone (EF24), a molecule having structural similarity to curcumin, was recently reported to inhibit proliferation of various cancer cells significantly. Here we try to determine the effect and mechanism of EF24 on hepatocellular carcinoma. 2 µM EF24 was found to inhibit the proliferation of PLC/PRF/5, Hep3B, HepG2, SK-HEP-1 and Huh 7 cell lines. However, even 8 µM EF24 treatment did not affect the proliferation of normal liver LO2 cells. Accordingly, 20 mg/kg/d EF24 inhibited the growth of the tumor xenografts conspicuously while causing no apparent change in liver, spleen or body weight. In addition, significant apoptosis and G2/M phase cell cycle arrest were found using flow cytometry. Besides, caspases and PARP activation and features typical of apoptosis including fragmented nuclei with condensed chromatin were also observed. Furthermore, the mechanism was targeted at the reduction of nuclear factor kappa b (NF-κB) pathway and the NF-κB–regulated gene products Bcl-2, COX-2, Cyclin B1. Our study has offered a strategy that EF24 being a therapeutic agent for hepatocellular carcinoma

Determination of hydroxyl groups in biorefinery resources via quantitative 31P NMR spectroscopy

The analysis of chemical structural characteristics of biorefinery product streams (such as lignin and tannin) has advanced substantially over the past decade, with traditional wet-chemical techniques being replaced or supplemented by NMR methodologies. Quantitative 31P NMR spectroscopy is a promising technique for the analysis of hydroxyl groups because of its unique characterization capability and broad potential applicability across the biorefinery research community. This protocol describes procedures for (i) the preparation/solubilization of lignin and tannin, (ii) the phosphitylation of their hydroxyl groups, (iii) NMR acquisition details, and (iv) the ensuing data analyses and means to precisely calculate the content of the different types of hydroxyl groups. Compared with traditional wet-chemical techniques, the technique of quantitative 31P NMR spectroscopy offers unique advantages in measuring hydroxyl groups in a single spectrum with high signal resolution. The method provides complete quantitative information about the hydroxyl groups with small amounts of sample (~30 mg) within a relatively short experimental time (~30-120 min)

Assessing the effect of pretreatment on cellulose accessibility for cellulosic biofuels production

Biomass recalcitrance has been recognized as one of the major barriers that hided the cost-effective conversion of lignocellulosic biomass to bioethanol, therefore the current bioconversion process require an essential step known as pretreatment to increase the cellulose accessibility. This thesis provides information about changes in cellulose accessibility upon different pretreatments, along with how these pretreatments alter the chemical and physical structures of biomass, will be extremely helpful to further optimize the current pretreatment process. Multiple promising analytical techniques including Simons’ stain, NMR cryoporometry, relaxometry, mercury porosimetry was introduced and successfully applied on pretreated biomass samples to characterize the cellulose accessible surface area and biomass porosity. Different pretreatments increase cellulose accessibility through different mechanisms to different extent. Dilute acid pretreatment is more effective than steam explosion in terms of increasing accessible surface area of cellulose as reflected by Simons’ stain and NMR cryoporometry, while NMR relaxometry suggested steam explosion is more effective at pore expansion for the cell wall water pools detected by changes in NMR relaxation time. Alkaline pretreatment decreased cellulose degree of polymerization, cellulose crystallinity, lignin content and subsequently increased cellulose accessibility, with sodium hydroxide pretreatment proved to be much more effective compared lime or soaking in ammonia pretreatment. Delignification through alkaline-based pretreatment is found less effective than removal of hemicellulose using acid in terms of cellulose accessibility increase. Lignin didn’t directly dictate cellulose accessibility but rather restricted xylan accessibility which in turn controls the access of cellulase to cellulose. Pore size distribution analysis based on mercury porosimetry also indicated that the most fundamental barrier in terms of biomass porosity scale for efficient enzymatic hydrolysis is the nano-pore space formed between coated microfibrils, despite some of the porous architecture such as cell lumen and pit could be severely destroyed after pretreatment. The action of cellulase on the characteristics of cellulosic fractions obtained from pretreated biomass was also investigated. Cellulose accessibility was found to increase at the beginning of hydrolysis, and after reaching a maximum value then starting to decrease. Enzymatic hydrolysis resulted in a rapid decrease in the cellulose degree of polymerization then gradually leveled off, suggesting the existence of a synergistic action of endo- and exo-glucanases that contribute to the occurrence of a peeling off type mechanism.Ph.D

A Site of Nation: Black Utopian Novels in the Late 19th and Early 20th Centuries

ABSTRACT Contrary to the traditional view that there is lack of utopian dimension in African American literature, this dissertation argues that African American literature not only develops an exuberant utopian tradition, but also forms its own utopian uniqueness. Based on this conclusion, the dissertation specially focuses on the period between the 1880s and the first two decades of the 20th century that witnessed the first peak of African American utopian writing. Meanwhile, this era has been claimed as the “Golden Age” of Black Nationalism. Through the examination of the historical background of the co-existence of these two conflicting strains, I contend that it not only provides fertile ground for the blooming of the utopian genre in African American literary writing, but also helps justify its popularity among African American writers. Through the analysis of three African American utopian writers: Sutton E. Griggs, Pauline Hopkins and W.E.B. Du Bois, I conclude that these writers use utopian texts to express their nation consciousness — and by so doing, challenged the myth of (a) white supremacy; (b) that African Americans are incapable of imagining an ideal world; and (c) carved a way forward for the black community