INVESTIGATION OF THE STRUCTURE, CHEMISTRY AND FUNCTIONAL PERFORMANCE OF NOVEL BIOCHARS

Biochars are a class of carbonaceous materials possessing high degrees of structural and chemical disorder in both organic and inorganic constituent phases. Despite this disorder, and in some cases because of this disorder, biochars have shown strong performance in adsorbing and even sequestering contaminants from soil, surface water and air. Biochar is the carbonaceous, solid product of heating carbonaceous feedstock in an oxygen-limited environment above 250°C, usually sourced from inexpensive, locally available agricultural and forestry wastes which can otherwise be difficult and expensive to dispose of. Biochar possesses attractive functional properties like high specific surface area, high micropore volume, and tunable surface chemistry which are key to performance in separation technologies and environmental remediation. In this work, two high-performing biochars were investigated: one from cottonwood feedstock and one sourced from locally available bovine bone waste. In the wood-based biochar, the synergistic impacts of potassium content and lignin microstructure were investigated for their role in determining biochar structure and function. The performance of these biochars was tested in dynamic adsorption of gaseous ammonia, a growing threat to human and environmental health, in which all biochars outperformed a commercial activated carbon. Breakthrough times showed no correlation with surface area of the adsorbents, contrary to expectation. Biochar sourced from pyrolyzed bovine bone has shown strong performance against a range of other materials in removing Cu2+ from acid mine waste. In-depth characterization of the mineral and organic phases of this novel biochar revealed that disorder may be key to its strong performance in Cu2+ removal

The Influence of Enterprise Systems on Business and Information Technology

Business strategy is important to all organizations. Nearly all Fortune 500 firms are implementing Enterprise Resource Planning (ERP) systems to improve the execution of their business strategy and to improve integration with its information technology (IT) strategy. Successful implementation of these multi-million dollar software systems are requiring new emphasis on change management and on Business and IT strategic alignment. This paper examines business and IT strategic alignment and seeks to explore whether an ERP implementation can drive business process reengineering and business and IT strategic alignment. An overview of business strategy and strategic alignment are followed by an analysis of ERP. The “As-Is/To-Be” process model is then presented and explained as a simple, but vital tool for improving business strategy, strategic alignment, and ERP implementation success

Dual-mode of insulin action controls GLUT4 vesicle exocytosis

Insulin releases an intracellular brake and promotes fusion pore expansion to translocate GLUT4 vesicles, and switches vesicle trafficking between distinct exocytic circuits

The divergent mitotic kinesin MKLP2 exhibits atypical structure and mechanochemistry

MKLP2, a kinesin-6, has critical roles during the metaphase-anaphase transition and cytokinesis. Its motor domain contains conserved nucleotide binding motifs, but is divergent in sequence (~35% identity) and size (~40% larger) compared to other kinesins. Using cryo-electron microscopy and biophysical assays, we have undertaken a mechanochemical dissection of the microtubule-bound MKLP2 motor domain during its ATPase cycle, and show that many facets of its mechanism are distinct from other kinesins. While the MKLP2 neck-linker is directed towards the microtubule plus-end in an ATP-like state, it does not fully dock along the motor domain. Furthermore, the footprint of the MKLP2 motor domain on the MT surface is altered compared to motile kinesins, and enhanced by kinesin-6-specific sequences. The conformation of the highly extended loop6 insertion characteristic of kinesin-6s is nucleotide-independent and does not contact the MT surface. Our results emphasize the role of family-specific insertions in modulating kinesin motor function

Observing GLUT4 Translocation in Live L6 Cells Using Quantum Dots

The glucose transporter 4 (GLUT4) plays a key role in maintaining whole body glucose homeostasis. Tracking GLUT4 in space and time can provide new insights for understanding the mechanisms of insulin-regulated GLUT4 translocation. Organic dyes and fluorescent proteins were used in previous studies for investigating the traffic of GLUT4 in skeletal muscle cells and adipocytes. Because of their relative weak fluorescent signal against strong cellular autofluorescence background and their fast photobleaching rate, most studies only focused on particular segments of GLUT4 traffic. In this study, we have developed a new method for observing the translocation of GLUT4 targeted with photostable and bright quantum dots (QDs) in live L6 cells. QDs were targeted to GLUT4myc specifically and internalized with GLUT4myc through receptor-mediated endocytosis. Compared with traditional fluorescence dyes and fluorescent proteins, QDs with high brightness and extremely photostability are suitable for long-term single particle tracking, so individual GLUT4-QD complex can be easily detected and tracked for long periods of time. This newly described method will be a powerful tool for observing the translocation of GLUT4 in live L6 cells

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MKLP2, a kinesin-6, has critical roles during the metaphase-anaphase transition and cytokinesis. Its motor domain contains conserved nucleotide binding motifs, but is divergent in sequence (~35% identity) and size (~40% larger) compared to other kinesins. Using cryo-electron microscopy and biophysical assays, we have undertaken a mechanochemical dissection of the microtubule-bound MKLP2 motor domain during its ATPase cycle, and show that many facets of its mechanism are distinct from other kinesins. While the MKLP2 neck-linker is directed towards the microtubule plus-end in an ATP-like state, it does not fully dock along the motor domain. Furthermore, the footprint of the MKLP2 motor domain on the MT surface is altered compared to motile kinesins, and enhanced by kinesin-6-specific sequences. The conformation of the highly extended loop6 insertion characteristic of kinesin-6s is nucleotide-independent and does not contact the MT surface. Our results emphasize the role of family-specific insertions in modulating kinesin motor function

Analysis of the kinetics of binding of Protein Kinase A Inhibitor alpha (PKIa) to cAMP-dependent protein kinase a catalytic subunit (PKA-C)

In the excel file are summarized the TR-FRET raw data used for the analysis of the binding kinetic for PKIa to ATP-saturated PKA-C. All the experiments were acquired at Biophysical Technology Center (BMBB Department, University of Minnesota, Minneapolis, MN) by GL and JM within The analysis of the TR-FRET data was performed by GL and JM.TR-FRET raw data used for the analysis of the binding kinetic for full-length protein kinase inhibitor (PKIa) to ATP-saturated cAMP-dependent protein kinase A (PKA-C). The experiments are part of a publication on eLIFE: "Multi-state Recognition Pathway of the Intrinsically Disordered Protein Kinase Inhibitor by Protein Kinase A", where we investigated the structural and kinetics changed that PKIa undergoes upon interaction with PKA-CNIH GM 100310 to G.

Loop L5 assumes three distinct orientations during the ATpase cycle of the mitotic kinesin eg5: a transient and time-resolved flourescence study

Members of the kinesin superfamily of molecular motors differ in several key structural domains, which probably allows these molecular motors to serve the different physiologies required of them. One of the most variable of these is a stem-loop motif referred to as L5. This loop is longest in the mitotic kinesin Eg5, and previous structural studies have shown that it can assume different conformations in different nucleotide states. However, enzymatic domains often consist of a mixture of conformations whose distribution shifts in response to substrate binding or product release, and this information is not available from the "static" images that structural studies provide. We have addressed this issue in the case of Eg5 by attaching a fluorescent probe to L5 and examining its fluorescence, using both steady state and time-resolved methods. This reveals that L5 assumes an equilibrium mixture of three orientations that differ in their local environment and segmental mobility. Combining these studies with transient state kinetics demonstrates that there is a major shift in this distribution during transitions that interconvert weak and strong microtubule binding states. Finally, in conjunction with previous cryo-EM reconstructions of Eg5·microtubule complexes, these fluorescence studies suggest a model in which L5 regulates both nucleotide and microtubule binding through a set of reversible interactions with helix α3. We propose that these features facilitate the production of sustained opposing force by Eg5, which underlies its role in supporting formation of a bipolar spindle in mitosis