Recombinant cellulose synthase

The present invention relates to the compositions and methods associated with the cloning of the catalytic subunit of cellulose synthase responsible for catalyzing cellulose biosynthesis. The invention relates further to compositions and methods for obtaining host cells containing recombinant cellulose synthase as well as compositions and methods for obtaining cellulose synthase from natural sources. In certain aspects, the present invention provides methods for the cloning of an 83 kd subunit of the cellulose synthase enzyme from Acetobacter xylinum.Board of Regents, University of Texas Syste

Modification of cellulose normally synthesizied by cellulose-producing microorganisms

The present invention involves a process for screening for and isolating spontaneously occurring or induced cellulose II-producing microorganisms. The process comprises a series of steps in various embodiments. Initially, cellulose-producing microorganisms from a first culture are plated out on a nutrient agar plate. The nutrient agar plate is then incubated to facilitate formation of colonies from single microorganisms. Samples of liquid nutrient medium are then inoculated with microorganisms from colonies having a smooth configuration, as compared to the usual rough colony configuration. The inoculated samples are then aerobically incubated to facilitate microorganism proliferation and pellicle formation. From these incubated samples are selected microorganisms, which, after a cultivation period, have proliferated but not formed a pellicle. Said selected microorganisms produce cellulose II instead of the cellulose I produced by pellicle-forming organisms.Board of Regents, University of Texas Syste

Social Studies Teachers’ Perceptions of Alternative Assessments for Students with IEPs

Assessments are one of the vital pillars of education but often are designed without the specific needs of students considered. Instruction is often differentiated for students with individualized education programs (IEPs), but rarely are assessments. Students in classes with IEPs in the social studies department at the project site school in the state of New York are performing below grade level. Teachers at this school are encouraged by the department chair to structure their assessments based on the Regents exam, which uses traditional assessment methods. The purpose of this case study was to investigate the perceptions of teachers regarding the implementation and use of alternative assessments to support students’ academic achievement in social studies classes. Guided by Davidson’s theory of interpretivism, data were gathered from interviews with seven social studies teachers regarding their perceptions of the use of alternative assessments to support students’ academic achievement in social studies classes and to support test preparation for New York state Regents exams. Data were also collected through document review of teachers’ lesson plans. Data analysis indicated that most teachers feel unprepared to use alternative assessments and teachers lack training or familiarity with alternative assessments. A professional development plan was created and implemented to train teachers on the design and implementation of alternative assessments with the goal of improving outcomes for students with IEPs. The results of this study have potential implications for positive social change that include increased use of alternative assessments at the local site to the benefit of students with IEPs

Cellulose Synthesizing Complexes in Vascular Plants and Procaryotes

Continuing the work initiated under DE-FG03-94ER20145, the following major accomplishments were achieved under DE-FG02-03ER15396 from 2003-2007: (a) we purified the acsD gene product of the Acetobacter cellulose synthase operon as well as transferred the CesA cellulose gene from Gossypium into E. coli in an attempt to crystallize this protein for x-ray diffraction structural analysis; however, crystallization attempts proved unsuccessful; (b) the Acetobacter cellulose synthase operon was successfully incorporated into Synechococcus, a cyanobacterium2; (c) this operon in Synechococcus was functionally expressed; (d) we successfully immunolabeled Vigna cellulose and callose synthase components and mapped their distribution before and after wounding; (e) we developed a novel method to produce replicas of cellulose synthases in tobacco BY-2 cells, and we demonstrated the cytoplasmic domain of the rosette TC; (f) from the moss Physcomitrella, we isolated two full-length cDNA sequences of cellulose synthase (PpCesA1 and PpCesA2) and attempted to obtain full genomic DNA sequences; (g) we examined the detailed molecular structure of a new form of non-crystalline cellulose known as nematic ordered cellulose (=NOC)3

The importance of microtubule-dependent tension in accurate chromosome segregation

Accurate chromosome segregation is vital for cell and organismal viability. The mitotic spindle, a bipolar macromolecular machine composed largely of dynamic microtubules, is responsible for chromosome segregation during each cell replication cycle. Prior to anaphase, a bipolar metaphase spindle must be formed in which each pair of chromatids is attached to microtubules from opposite spindle poles. In this bipolar configuration pulling forces from the dynamic microtubules can generate tension across the sister kinetochores. The tension status acts as a signal that can destabilize aberrant kinetochore-microtubule attachments and reinforces correct, bipolar connections. Historically it has been challenging to isolate the specific role of tension in mitotic processes due to the interdependency of attachment and tension status at kinetochores. Recent technical and experimental advances have revealed new insights into how tension functions during mitosis. Here we summarize the evidence that tension serves as a biophysical signal that unifies multiple aspects of kinetochore and centromere function to ensure accurate chromosome segregation

Differentiating between models of Epothilone binding to microtubules using tubulin mutagenesis, cytotoxicity, and molecular modeling

This is the peer reviewed version of the following article: Entwistle, R. A., Rizk, R. S., Cheng, D. M., Lushington, G. H., Himes, R. H., & Gupta, M. L. (2012). Differentiating between models of Epothilone binding to microtubules using tubulin mutagenesis, cytotoxicity, and molecular modeling. ChemMedChem, 7(9), 1580–1586. http://doi.org/10.1002/cmdc.201200286, which has been published in final form at doi.org/10.1002/cmdc.201200286. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.Microtubule stabilizers are powerful anti-mitotic compounds and represent a proven cancer treatment strategy. Several classes of compounds in clinical use or trials, such as the taxanes and epothilones, bind to the same region of β-tubulin. Determining how these molecules interact with tubulin and stabilize microtubules is important both for understanding the mechanism of action and enhancing chemotherapeutic potential, e.g. reducing side effects, increasing solubility, and overcoming resistance. Structural studies using nonpolymerized tubulin or stabilized polymers have produced different models of epothilone binding. Here, we used directed mutagenesis of the binding site on Saccharomyces cerevisiae β-tubulin to analyze interactions between Epothilone B and its biologically relevant substrate, dynamic microtubules. Five engineered amino acid changes contributed to a 125-fold increase in Epothilone B cytotoxicity independent of inherent microtubule stability. The mutagenesis of endogenous β-tubulin was done in otherwise isogenic strains. This facilitated the correlation of amino acid substitutions with altered cytotoxicity using molecular mechanics simulations. The results, which are based on the interaction between Epothilone B and dynamic microtubules, most strongly support the binding mode determined by NMR spectroscopy-based studies. This work establishes a system for discriminating between potential binding modes and among various compounds and/or analogues using a sensitive biological activity-based readout

Analysis of Stainless Steel Sandwich Panels with a Metal Foam Care for Lightweight Fan Blade Design

The quest for cheap, low density and high performance materials in the design of aircraft and rotorcraft engine fan and propeller blades poses immense challenges to the materials and structural design engineers. Traditionally, these components have been fabricated using expensive materials such as light weight titanium alloys, polymeric composite materials and carbon-carbon composites. The present study investigates the use of P sandwich foam fan blade made up of solid face sheets and a metal foam core. The face sheets and the metal foam core material were an aerospace grade precipitation hardened 17-4 PH stainless steel with high strength and high toughness. The stiffness of the sandwich structure is increased by separating the two face sheets by a foam core. The resulting structure possesses a high stiffness while being lighter than a similar solid construction. Since the face sheets carry the applied bending loads, the sandwich architecture is a viable engineering concept. The material properties of 17-4 PH metal foam are reviewed briefly to describe the characteristics of the sandwich structure for a fan blade application. A vibration analysis for natural frequencies and P detailed stress analysis on the 17-4 PH sandwich foam blade design for different combinations of skin thickness and core volume %re presented with a comparison to a solid titanium blade

An extragalactic supernebula confined by gravity

Little is known about the origins of the giant star clusters known as globular clusters. How can hundreds of thousands of stars form simultaneously in a volume only a few light years across the distance of the sun to its nearest neighbor? Radiation pressure and winds from luminous young stars should disperse the star-forming gas and disrupt the formation of the cluster. Globular clusters in our Galaxy cannot provide answers; they are billions of years old. Here we report the measurement of infrared hydrogen recombination lines from a young, forming super star cluster in the dwarf galaxy, NGC 5253. The lines arise in gas heated by a cluster of an estimated million stars, so young that it is still enshrouded in gas and dust, hidden from optical view. We verify that the cluster contains 4000-6000 massive, hot "O" stars. Our discovery that the gases within the cluster are bound by gravity may explain why these windy and luminous O stars have not yet blown away the gases to allow the cluster to emerge from its birth cocoon. Young clusters in "starbursting" galaxies in the local and distant universe may be similarly gravitationally confined and cloaked from view.Comment: Letter to Natur