Open Access. Powered by Scholars. Published by Universities.®

Biochemistry, Biophysics, and Structural Biology Commons

Open Access. Powered by Scholars. Published by Universities.®

Articles 1 - 12 of 12

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Characterization Of A Recently Purified Thermophilic Dnase From A Novel Thermophilic Fungus, Kyle Landry, Robert Levin Jul 2014

Characterization Of A Recently Purified Thermophilic Dnase From A Novel Thermophilic Fungus, Kyle Landry, Robert Levin

Kyle S Landry

A newly isolated thermophilic fungus was found to produce a partially inducible extracellular DNase. This manuscript focuses on the characterization of this novel thermophilic DNase in terms of optimal enzyme conditions, molecular weight, and certain kinetic properties. The DNase was found to be inactivated by the presence of EDTA demonstrating its dependence on metal cofactors for activity. Maximum activity occurred at pH 6.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the purified DNase was 65 °C. The thermophilic DNase was found to be an exonuclease with an estimated molecular weight of 56 ...


Development Of A Novel Affinity Membrane Purification System For Deoxyribonuclease, Kyle Landry, Robert Levin Jan 2014

Development Of A Novel Affinity Membrane Purification System For Deoxyribonuclease, Kyle Landry, Robert Levin

Kyle S Landry

A membrane based affinity purification system was developed for the purification of the DNA specific nuclease, DNase I. Single stranded DNA was bound to unmodified polyvinylidene fluoride (PVDF) membranes which were used to purify DNase I from a solution of bovine serum albumin. Using coated membranes, a 6-fold increase in specific activity was achieved with 80 % enzyme recovery. This method provides a simple yet effective way to purify DNase I and can be very useful for the purification of other DNA specific enzymes.


Purification Of An Inducible Dnase From A Thermophilic Fungus, Kyle Landry, Andrea Vu, Robert Levin Dec 2013

Purification Of An Inducible Dnase From A Thermophilic Fungus, Kyle Landry, Andrea Vu, Robert Levin

Kyle S Landry

The ability to induce an extracellular DNase from a novel thermophilic fungus was studied and the DNAse purified using both traditional and innovative purification techniques. The isolate produced sterile hyphae under all attempted growing conditions, with an average diameter of 2 μm and was found to have an optimal temperature of 45 °C and a maximum of 65 °C. Sequencing of the internal transcribed region resulted in a 91% match with Chaetomium sp., suggesting a new species, but further clarification on this point is needed. The optimal temperature for DNase production was found to be 55 °C and was induced ...


Purification And Characterization Of Iso-Ribonucleases From A Novel Thermophilic Fungus, Kyle Landry, Robert Levin Dec 2013

Purification And Characterization Of Iso-Ribonucleases From A Novel Thermophilic Fungus, Kyle Landry, Robert Levin

Kyle S Landry

A thermophilic fungus previously isolated from composted horse manure was found to produce extracellular iso-RNases that were purified 127.6-fold using a combination of size exclusion chromatography and a novel affinity membrane purification system. The extent of purification was determined electrophoretically using 4%–15% gradient polyacrylamide gels. RNase activity was dependent on the presence of a metal co-factor with significantly more activity with Zn2+ or Mn2+ than Mg2+. The RNases exhibited maximum activity at both pH 3.0 and pH 7.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the ...


Acute Toxicity Of Copper Sulfate And Potassium Dichromate On Stygobiont Proasellus: General Aspects Of Groundwater Ecotoxicology And Future Perspectives, Ana Reboleira, Nelson Abrantes, Pedro Oromí, Fernando Gonçalves Mar 2013

Acute Toxicity Of Copper Sulfate And Potassium Dichromate On Stygobiont Proasellus: General Aspects Of Groundwater Ecotoxicology And Future Perspectives, Ana Reboleira, Nelson Abrantes, Pedro Oromí, Fernando Gonçalves

Ana Sofia P.S. Reboleira

Karst systems harbor large groundwater resources for human consumption and represent an important habitat for rare and unprotected specialized animals, the so-called stygofauna. Due to the highly adapted features towards underground life, together with the geographic isolation provided by the subterranean aquifers, groundwater-dwelling animals may lose the ability to face sudden changes on their ecosystems, and therefore the risk of extinction is remarkably high. A little is known about their sensitiveness, especially linked to contamination pressure in urbanized karst areas. Understanding the impact of contaminants on stygofauna is important for setting groundwater environmental quality and management of karst systems. We ...


Purification And Characterization Of Novel Nucleases From A Thermophilic Fungus, Kyle Landry Sep 2012

Purification And Characterization Of Novel Nucleases From A Thermophilic Fungus, Kyle Landry

Kyle S Landry

A thermophilic fungus was isolated from composted horse manure. The organism was as a Chaetomium sp. by sequencing the highly conserved ITS region of the fungus and comparing to known regions in a genomic database and was referred to as TM-417. TM-417 was found to have an optimal growth temperature of 45 oC and an optimal pH of 7.0. An extracellular DNase and RNase was found to be produced by the isolate and were purified 145.58-fold and 127.6-fold respectively using a combination of size exclusion chromatography and a novel affinity membrane purification system. The extent of ...


Viability Of Lactic Acid Bacteria And Sensory Evaluation In Cinnamomum Verum And Allium Sativum-Bio-Yogurts Made From Camel And Cow Milk, Ahmad Salihin Hj Baba, Shori A. B. Dec 2011

Viability Of Lactic Acid Bacteria And Sensory Evaluation In Cinnamomum Verum And Allium Sativum-Bio-Yogurts Made From Camel And Cow Milk, Ahmad Salihin Hj Baba, Shori A. B.

Ahmad Salihin Hj Baba

The present study investigate the effect of herbal water extract prepared from Allium sativum and Cinnamomum verum on the viability of lactic acid bacteria (Lactobacillus spp and Streptococcus thermophilus) in cow- and camel-milk yogurts during 21 day refrigerated storage. The organoleptic properties of fresh-yogurts were evaluated. Lactobacillus spp count for fresh cow milk-yogurts (0 day) in both present and absent of C. verum and A. sativum was ranged from 1.4×0 6 to 2.1×10 6 cfu/mL. These values were not significantly changed throughout the 21 days of refrigerated storage. Lactobacillusspp count in fresh plain camel milk- ...


Confocal Laser Scanning Microscopy: A Flexible Tool For Polarized Light And Three-Dimensional Fluorescence Imaging Of Compact Bone, Corey Maggiano, Tosha Dupras, Michael Schultz, John Biggerstaff Sep 2009

Confocal Laser Scanning Microscopy: A Flexible Tool For Polarized Light And Three-Dimensional Fluorescence Imaging Of Compact Bone, Corey Maggiano, Tosha Dupras, Michael Schultz, John Biggerstaff

John Biggerstaff

Wide-field polarized light and epifluorescence microscopy have been used to enhance analysis of archaeological bone tissue, providing information on bone formation, modeling, pathology, preservation, age estimation, and biomechanics. Though valuable, these techniques are limited by their inability to remove out-of-focus light and view multiple levels of a sample, restricting our understanding of the three-dimensional (3-D) microarchitecture of compact bone. Modern technological advances, such as microscopic computerized tomography, allow increasing resolution in 3-D bone imaging, but do not allow fluorescence labeling or polarized-light analysis. Confocal laser scanning microscopy (CLSM) is a valuable tool for 3-D histology. However, its application to the ...


Synthesis, Biological Evaluation And Radiochemical Labeling Of A Dansylhydrazone Derivative As A Potential Imaging Agent For Apoptosis, Wenbin Zeng, Min-Liang Yao, David Townsend, George Kabalka, Jonathan Wall, Michael Le Puil, John Biggerstaff, Weimin Miao Jun 2008

Synthesis, Biological Evaluation And Radiochemical Labeling Of A Dansylhydrazone Derivative As A Potential Imaging Agent For Apoptosis, Wenbin Zeng, Min-Liang Yao, David Townsend, George Kabalka, Jonathan Wall, Michael Le Puil, John Biggerstaff, Weimin Miao

John Biggerstaff

To develop a small molecule-based tracer for in vivo apoptosis imaging, dansylhydrazone (DFNSH) was synthesized in 93% yield in less than 30 min. The biological evaluation showed that DFNSH selectively binds to paclitaxel-induced apoptotic cancer cells. The high magnification fluorescent images demonstrate that DFNSH is localized within the cytoplasm of cells that bound Alexa® 488 labeled annexin V on the plasma membrane. [18F]-DFNSH ([18F]-3) was synthesized and isolated in 50–60% radiochemical yields, based on [K/K222]18F, with a synthesis time of 50 min (EOB). The straightforward preparation of fluorine-18 labeled 3 makes it a promising tracer ...


Soluble Fibrin Inhibits Lymphocyte Adherence And Cytotoxicity Against Tumor Cells: Implications For Cancer Metastasis And Immunotherapy, John Biggerstaff, Brandy Weidow, Judith Dexheimer, Gary Warnes, Jacqueline Vidosh, Shonak Patel, Michael Newman, Pretesh Patel Mar 2008

Soluble Fibrin Inhibits Lymphocyte Adherence And Cytotoxicity Against Tumor Cells: Implications For Cancer Metastasis And Immunotherapy, John Biggerstaff, Brandy Weidow, Judith Dexheimer, Gary Warnes, Jacqueline Vidosh, Shonak Patel, Michael Newman, Pretesh Patel

John Biggerstaff

Circulating soluble fibrin (sFn) is elevated in many cancer patients. It is a marker for ongoing disseminated intravascular coagulation and may have prognostic significance. We have demonstrated that sFn inhibited monocyte adherence and cytotoxicity by a mechanism involving blockade of monocyte αMβ2 and tumor cell CD54. It was, therefore, hypothesized that sFn also inhibits lymphocyte and interleukin-2—activated lymphocyte (LAK) adherence and cytotoxicity against tumor cells. This study sought to identify the lymphocyte subset responsible for adherence and killing of A375 melanoma cells and whether sFn inhibited these parameters. Lymphocyte and LAK cell adherence and cytotoxicity, which was adherence dependent ...


Micropropagation Of Field-Grown Perennial Teosinte From Node Culture, J. Zale, L. Ohnoutková, D. West, C. Sams, John Biggerstaff, M. Le Puil Dec 2007

Micropropagation Of Field-Grown Perennial Teosinte From Node Culture, J. Zale, L. Ohnoutková, D. West, C. Sams, John Biggerstaff, M. Le Puil

John Biggerstaff

Propagation of perennial diploid teosinte, Zea diploperennis Iltis, Doebley and Guzmán (2n = 2x = 20), and tetraploid teosinte, Z. perennis (Hitchc.) Reeves and Magelsdorf (2n = 4x = 40), is limited by seed availability. For plants grown in the field, a photoperiod response delays flowering until fall, and seeds may not reach maturity. The objective of this study was to develop an in vitro micropropagation protocol for field grown teosinte. Shoot proliferation was induced from nodes split longitudinally and plated on Murashige and Skoog's (MS) medium supplemented with 5 μM 6-benzyl amino purine and 3% (w/v) sucrose. Both species possess a ...


Growth And Reproductive Performance Of Small Ruminants Under Integrated Livestock Oil Palm Production System, Ahmad Salihin Hj Baba, Azillah A., Mukherjee T. K., Abdullah R. B. Dec 1997

Growth And Reproductive Performance Of Small Ruminants Under Integrated Livestock Oil Palm Production System, Ahmad Salihin Hj Baba, Azillah A., Mukherjee T. K., Abdullah R. B.

Ahmad Salihin Hj Baba

The effects of supplementation with mixed fodder or concentrate were investigated on the performance of a) growth of male lambs;md kids grazing extensively under oil palm trees and b) reproduction of ewes and goats stall-fed with native herbage. Supplemented kids and lambs had higher average daily weight gain than controls but the effect was only significant for those which received concentrate (p < 0.05). Final body weight was only significantly different from controls for lambs supplemented with concentrate (p < 0.05). The ages at puberty, first mating, first conception and first kidding of supplemented goats were about 110 days earlier than those for controls (p < 0.05 or better). Supplemented goats had first mating and conceived at lower body weights (p < 0.01) than those in control groups. Different feeding regimes had no effects (p > 0.05) on the reproductive performance of ewes apart fi om highest body weight of first lambing in animals supplemented with concentrate (p < 0.05). Native herbage available under oil palm trees of 5 years old was ...