Simultaneous saccharification and fermentation of steam exploded duckweed: Improvement of the ethanol yield by increasing yeast titre

This study investigated the conversion of Lemna minor biomass to bioethanol. The biomass was pre-treated by steam explosion (SE, 210 °C, 10 min) and then subjected to simultaneous saccharification and fermentation (SSF) using CellicÒ CTec 2 (20 U or 0.87 FPU gﰂ1 substrate) cellulase plus b-glucosidase (2 U gﰂ1 substrate) and a yeast inoculum of 10% (v/v or 8.0 ﰀ 107 cells mLﰂ1). At a substrate concentration of 1% (w/v) an ethanol yield of 80% (w/w, theoretical) was achieved. However at a substrate concentration of 20% (w/v), the ethanol yield was lowered to 18.8% (w/w, theoretical). Yields were considerably improved by increasing the yeast titre in the inoculum or preconditioning the yeast on steam exploded liquor. These approaches enhanced the ethanol yield up to 70% (w/w, theoretical) at a substrate concen- tration of 20% (w/v) by metabolising fermentation inhibitors

The Transcription Factor Rfx3 Regulates β-Cell Differentiation, Function, and Glucokinase Expression

OBJECTIVE: Pancreatic islets of perinatal mice lacking the transcription factor Rfx3 exhibit a marked reduction in insulin-producing beta-cells. The objective of this work was to unravel the cellular and molecular mechanisms underlying this deficiency. RESEARCH DESIGN AND METHODS: Immunofluorescence studies and quantitative RT-PCR experiments were used to study the emergence of insulin-positive cells, the expression of transcription factors implicated in the differentiation of beta-cells from endocrine progenitors, and the expression of mature beta-cell markers during development in Rfx3(-/-) and pancreas-specific Rfx3-knockout mice. RNA interference experiments were performed to document the consequences of downregulating Rfx3 expression in Min6 beta-cells. Quantitative chromatin immunoprecipitation (ChIP), ChIP sequencing, and bandshift experiments were used to identify Rfx3 target genes. RESULTS: Reduced development of insulin-positive cells in Rfx3(-/-) mice was not due to deficiencies in endocrine progenitors or beta-lineage specification, but reflected the accumulation of insulin-positive beta-cell precursors and defective beta-cells exhibiting reduced insulin, Glut-2, and Gck expression. Similar incompletely differentiated beta-cells developed in pancreas-specific Rfx3-deficient embryos. Defective beta-cells lacking Glut-2 and Gck expression dominate in Rfx3-deficent adults, leading to glucose intolerance. Attenuated Glut-2 and glucokinase expression, and impaired glucose-stimulated insulin secretion, were also induced by RNA interference-mediated inhibition of Rfx3 expression in Min6 cells. Finally, Rfx3 was found to bind in Min6 cells and human islets to two well-known regulatory sequences, Pal-1 and Pal-2, in the neuroendocrine promoter of the glucokinase gene. CONCLUSIONS: Our results show that Rfx3 is required for the differentiation and function of mature beta-cells and regulates the beta-cell promoter of the glucokinase gene

Deregulation of CREB Signaling Pathway Induced by Chronic Hyperglycemia Downregulates NeuroD Transcription

CREB mediates the transcriptional effects of glucose and incretin hormones in insulin-target cells and insulin-producing β-cells. Although the inhibition of CREB activity is known to decrease the β-cell mass, it is still unknown what factors inversely alter the CREB signaling pathway in β-cells. Here, we show that β-cell dysfunctions occurring in chronic hyperglycemia are not caused by simple inhibition of CREB activity but rather by the persistent activation of CREB due to decreases in protein phophatase PP2A. When freshly isolated rat pancreatic islets were chronically exposed to 25 mM (high) glucose, the PP2A activity was reduced with a concomitant increase in active pCREB. Brief challenges with 15 mM glucose or 30 µM forskolin after 2 hour fasting further increased the level of pCREB and consequently induced the persistent expression of ICER. The excessively produced ICER was sufficient to repress the transcription of NeuroD, insulin, and SUR1 genes. In contrast, when islets were grown in 5 mM (low) glucose, CREB was transiently activated in response to glucose or forskolin stimuli. Thus, ICER expression was transient and insufficient to repress those target genes. Importantly, overexpression of PP2A reversed the adverse effects of chronic hyperglycemia and successfully restored the transient activation of CREB and ICER. Conversely, depletion of PP2A with siRNA was sufficient to disrupt the negative feedback regulation of CREB and induce hyperglycemic phenotypes even under low glucose conditions. Our findings suggest that the failure of the negative feedback regulation of CREB is the primary cause for β-cell dysfunctions under conditions of pathogenic hyperglycemia, and PP2A can be a novel target for future therapies aiming to protect β-cells mass in the late transitional phase of non-insulin dependent type 2 diabetes (NIDDM)

Shipwrecks and sports divers: Florida's programs in participatory preservation underwater

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Enzymatic saccharification of duckweed (Lemna minor) biomass without thermophysical pretreatment

Duckweed is a rapidly replicating aquatic plant that has the potential to decontaminate effluent streams from food processing and also has a low-lignin content. Hence it could provide a more suitable source of cellulose for conversion to biofuels. This paper reports that duckweed biomass has the potential to be enzymatically saccharified to produce glucose and other cell-wall-derived sugars which might be converted to ethanol by fermentation or exploited as industrial platform chemicals. The enzymatic digestibility has been studied on alcohol-extracted, water-insoluble preparations of duckweed cell walls. Within these, glucose accounts for w = 25.4% (dry wt), which has arisen from cellulose and non-cellulosic glucans including starch. Several commercial cell-wall degrading enzymes and cocktails have been evaluated. Saccharification can be achieved within about 8 h using commercial cellulase at 4.35 FPU g−1 substrate in conjunction with added beta-glucosidase at 100 U g−1 substrate. The potential for exploiting duckweed is discussed

Can qualitatively similar temperature-histories be obtained in different pilot HP units?

An experimental protocol to harmonize the pressure and temperature-histories of model samples treated in different individual high pressure pilot units was developed. This protocol was based on the endpoint strategy. Step zero of this protocol consisted of an exploratory measurement of the pressure, temperature-history of the pressure transmitting medium in the different chambers without the use of a container. In steps one and two of the protocol, the temperature of a sample was measured, which was placed in a container. Two types of samples were considered, a pure water sample allowing free convection (step 1) and a viscous water-based system (using a water soluble thickener) preventing free convection in the sample container (step 2). The high pressure units differed primarily in pressure build-up and pressure release times. The suggested endpoint strategy enabled the minimization of differences in temperature-histories during the pressure holding phase. Pressure, temperature-histories measured in the different high pressure units and information on the inactivation kinetics of a-amylase based systems were used to compare the process impact of different pressure, temperature-histories on the enzyme inactivation. The differences in temperature-histories significantly influenced the degree of enzyme inactivation; in particular the temperature-histories during the pressure build-up phase substantially contributed to differences in residual enzyme activity