Activated protein C protects against diabetic nephropathy by inhibiting endothelial and podocyte apoptosis

Data providing direct evidence for a causative link between endothelial dysfunction, microvascular disease and diabetic end-organ damage are scarce. Here we show that activated protein C (APC) formation, which is regulated by endothelial thrombomodulin, is reduced in diabetic mice and causally linked to nephropathy. Thrombomodulin-dependent APC formation mediates cytoprotection in diabetic nephropathy by inhibiting glomerular apoptosis. APC prevents glucose-induced apoptosis in endothelial cells and podocytes, the cellular components of the glomerular filtration barrier. APC modulates the mitochondrial apoptosis pathway via the protease-activated receptor PAR-1 and the endothelial protein C receptor EPCR in glucose-stressed cells. These experiments establish a new pathway, in which hyperglycemia impairs endothelial thrombomodulin-dependent APC formation. Loss of thrombomodulin-dependent APC formation interrupts cross-talk between the vascular compartment and podocytes, causing glomerular apoptosis and diabetic nephropathy. Conversely, maintaining high APC levels during long-term diabetes protects against diabetic nephropathy.     

Aqueous two phase extraction of invertase from baker's yeast: Effect of process parameters on partitioning

Invertase (β-d-fructofuronoside fructohydrolase) is an industrially important enzyme useful for the hydrolysis of sucrose. The potential of aqueous two phase extraction for the isolation and purification of invertase from crude baker's yeast is explored. Influence of the process parameters such as type of phase forming salts, PEG molecular weight, concentration of salt and polymer, tie line length and volume ratio on partitioning of invertase was studied. PEG 3350/magnesium sulphate system was found most suitable for the extraction which has resulted in favorable pH (5 ± 0.2) for the enzyme extraction. Polymer and salt concentration were found to significantly affect the degree of purification and enzyme recovery of invertase. The purity of ∼8.81 fold was obtained compared to crude extract with recovery of 77% at the standardized process conditions. Overall results demonstrated the feasibility of aqueous two phase extraction for the isolation and purification of invertase without the need of multiple steps.     

MapReduce framework energy adaptation via temperature awareness

MapReduce has become a popular framework for Big Data applications. While MapReduce has received much praise for its scalability and efficiency, it has not been thoroughly evaluated for power consumption. Our goal with this paper is to explore the possibility of scheduling in a power-efficient manner without the need for expensive power monitors on every node. We begin by considering that no cluster is truly homogeneous with respect to energy consumption. From there we develop a MapReduce framework that can evaluate the current status of each node and dynamically react to estimated power usage. In so doing, we shift work toward more energy efficient nodes which are currently consuming less power. Our work shows that given an ideal framework configuration, certain nodes may consume only 62.3 % of the dynamic power they consumed when the same framework was configured as it would be in a traditional MapReduce implementation.     

Discovery of bacterial NAD⁺-dependent DNA ligase inhibitors: improvements in clearance of adenosine series

Optimization of clearance of adenosine inhibitors of bacterial NAD(+)-dependent DNA ligase is discussed. To reduce Cytochrome P-450-mediated metabolic clearance, many strategies were explored; however, most modifications resulted in compounds with reduced antibacterial activity and/or unchanged total clearance. The alkyl side chains of the 2-cycloalkoxyadenosines were fluorinated, and compounds with moderate antibacterial activity and favorable pharmacokinetic properties in rat and dog were identified.     

Feedback inhibition of spinach L-galactose dehydrogenase by L-ascorbate

We have studied the enzymological properties of L-galactose dehydrogenase (l-GalDH), a key enzyme in the biosynthetic pathway of l-ascorbate (AsA) in plants. L-GalDH was purified approximately 560-fold from spinach leaves. The enzyme was a homodimer with a subunit mass of 36 kDa. We also cloned the full-length cDNA of spinach L-GalDH, which contained an open reading frame encoding 322 amino acid residues with a calculated molecular mass of 35,261 Da. The deduced amino acid sequence of the cDNA showed 82, 79 and 75% homology to L-GalDH from kiwifruit, apple and Arabidopsis, respectively. Recombinant enzyme expressed from the cDNA in Escherichia coli showed L-GalDH activity. Southern blot analysis revealed that the spinach L-GalDH gene occurs in a single copy. Northern blot analysis suggests that L-GalDH is expressed in different organs of spinach. The purified native L-GalDH showed high specificity for L-galactose with a Km of 116.2+/-3.2 microM. Interestingly, spinach L-GalDH exhibited reversible inhibition by AsA, the end-product of the biosynthetic pathway. The inhibition kinetics indicated a linear-competitive inhibition with a Ki of 133.2+/-7.2 microM, suggesting feedback regulation in AsA synthesis in the plant.     

Mating frequency in dispersing potato tuber moth, Phthorimaea operculella, and its influence on the design of refugia to manage resistance in Bt transgenic crops

Mating of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), was investigated in relation to the dispersal of males in laboratory and field trials. The effect of stimulating the flight of males to light sources in a large cage on their mating ability was estimated for three age groups, and compared with similar estimates for confined moths. Although the mating of males declined with ages of up to 15 days, simulated dispersal had no effect on subsequent mating when the males were paired with virgin females. The dispersal of male moths was also categorised by the initial flight activity of untethered moths to a light source. Scores for poor, moderate, and good flight provided a repeatable measure of initial male flight activity, but the degree of activity was not related to their subsequent mating ability. In the field, virgin female potato tuber moths were tethered at various distances from the edge of isolated potato crops and then dissected to determine their mating status. Female mating frequency averaged 75% at the crop margin, remained above 50% up to 200 m, and then declined to 19% at 360 m from the margin. Derivation of the mating probability for an individual male potato tuber moth confirmed earlier work by other researchers that has indicated a tendency for dispersal prior to mating, and that males retain their ability to mate as they disperse from a crop. The influence of dispersal and mating on gene flow between crops, and its potential effects on refuge size required to minimise the development of resistance to Bt transgenic potato crops was examined.