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Articles 1 - 16 of 16

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

Comparison Of Electrically Mediated And Liposome-Complexed Plasmid Dna Delivery To The Skin, Loree C. Heller, Mark J. Jaroszeski, Domenico Coppola, Richard Heller Dec 2008

Comparison Of Electrically Mediated And Liposome-Complexed Plasmid Dna Delivery To The Skin, Loree C. Heller, Mark J. Jaroszeski, Domenico Coppola, Richard Heller

Bioelectrics Publications

BACKGROUND: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery.

METHODS: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model.

RESULTS: Experiments confirm ...


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 ...


Treg Depletion Inhibits Efficacy Of Cancer Immunotherapy: Implications For Clinical Trials., James Curtin, Marianela Candolfi, Tamer Fakhouri, Chunyan Liu, Anderson Alden, Matthew Edwards, Pedro Lowenstein, Maria Castro Apr 2008

Treg Depletion Inhibits Efficacy Of Cancer Immunotherapy: Implications For Clinical Trials., James Curtin, Marianela Candolfi, Tamer Fakhouri, Chunyan Liu, Anderson Alden, Matthew Edwards, Pedro Lowenstein, Maria Castro

Articles

BACKGROUND: Regulatory T lymphocytes (Treg) infiltrate human glioblastoma (GBM); are involved in tumor progression and correlate with tumor grade. Transient elimination of Tregs using CD25 depleting antibodies (PC61) has been found to mediate GBM regression in preclinical models of brain tumors. Clinical trials that combine Treg depletion with tumor vaccination are underway to determine whether transient Treg depletion can enhance anti-tumor immune responses and improve long term survival in cancer patients. FINDINGS: Using a syngeneic intracrabial glioblastoma (GBM) mouse model we show that systemic depletion of Tregs 15 days after tumor implantation using PC61 resulted in a decrease in Tregs ...


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 ...


Turning The Gene Tap Off; Implications Of Regulating Gene Expression For Cancer Therapeutics, James Curtin, Marianela Candolfi, Weidong Xiong, Pedro Lowenstein, Maria Castro Mar 2008

Turning The Gene Tap Off; Implications Of Regulating Gene Expression For Cancer Therapeutics, James Curtin, Marianela Candolfi, Weidong Xiong, Pedro Lowenstein, Maria Castro

Articles

Cancer poses a tremendous therapeutic challenge worldwide, highlighting the critical need for developing novel therapeutics. A promising cancer treatment modality is gene therapy, which is a form of molecular medicine designed to introduce into target cells genetic material with therapeutic intent. Anticancer gene therapy strategies currently used in preclinical models, and in some cases in the clinic, include proapoptotic genes, oncolytic/replicative vectors, conditional cytotoxic approaches, inhibition of angiogenesis, inhibition of growth factor signaling, inactivation of oncogenes, inhibition of tumor invasion and stimulation of the immune system. The translation of these novel therapeutic modalities from the preclinical setting to the ...


Emergent Decision-Making In Biological Signal Transduction Networks, Tomáš Helikar, John Konvalina, Jack Heidel, Jim A. Rogers Feb 2008

Emergent Decision-Making In Biological Signal Transduction Networks, Tomáš Helikar, John Konvalina, Jack Heidel, Jim A. Rogers

Biochemistry -- Faculty Publications

The complexity of biochemical intracellular signal transduction networks has led to speculation that the high degree of interconnectivity that exists in these networks transforms them into an information processing network. To test this hypothesis directly, a large scale model was created with the logical mechanism of each node described completely to allow simulation and dynamical analysis. Exposing the network to tens of thousands of random combinations of inputs and analyzing the combined dynamics of multiple outputs revealed a robust system capable of clustering widely varying input combinations into equivalence classes of biologically relevant cellular responses. This capability was nontrivial in ...


Analogy Of Issr And Rapd Markers For Comparative Analysis Of Genetic Diversity Among Different Jatropha Curcas Genotypes, Shailesh K. Tiwari Dr. Jan 2008

Analogy Of Issr And Rapd Markers For Comparative Analysis Of Genetic Diversity Among Different Jatropha Curcas Genotypes, Shailesh K. Tiwari Dr.

Shailesh K Tiwari Dr.

The phylogenetic relationships of 13 Jatropha genotypes from different parts of the India were analysed using 34 polymerase chain reaction (PCR) markers (20 random amplified polymorphic DNAs (RAPDs) and 14 inter simple sequence repeats (ISSRs)). Amplification of genomic DNA of the 13 genotypes, using RAPD analysis, yielded 107 fragments that could be scored, of which 91 were polymorphic, with an average of 4.55 polymorphic fragments per primer. Number of amplified fragments ranged from one (OPA20, OPB19, OPD13) to nine (OPA18) and which varied in size from 200 to 2,500 bp. Percentage of polymorphism ranged from 40% (OPB18) to ...


A Kinetic Model For Growth And Biosynthesis Of Medium-Chain-Length Poly-(3-Hydroxyalkanoates) In Pseudomonas Putida, Irene Tan Kit Ping Jan 2008

A Kinetic Model For Growth And Biosynthesis Of Medium-Chain-Length Poly-(3-Hydroxyalkanoates) In Pseudomonas Putida, Irene Tan Kit Ping

Irene Tan Kit Ping

A kinetic model is presented giving a mathematical description of batch culture of Pseudomonas putida PGA1 grown using saponified palm kernel oil as carbon source and ammonium as the limiting nutrient. The growth of the micro-organism is well-described using. Tessier-type model which takes into account the inhibitory effect of ammonium at high concentrations. The ammonium consumption rate by the cells is related ill proportion to the rate of growth. The intracellular production of medium-chain-length poly(3-hydroxyalkanoates) (PHA(MCL)) by P. putida PGA1 cells is reasonably modeled by the modified Luedeking-Piret kinetics, which incorporate a function of product Synthesis inhibition (or ...


Coa2 Is An Assembly Factor For Yeast Cytochrome C Oxidase Biogenesis That Facilitates The Maturation Of Cox1, Fabien Pierrel, Oleh Khalimonchuk, Paul A. Cobine, Megan Bestwick, Dennis R. Winge Jan 2008

Coa2 Is An Assembly Factor For Yeast Cytochrome C Oxidase Biogenesis That Facilitates The Maturation Of Cox1, Fabien Pierrel, Oleh Khalimonchuk, Paul A. Cobine, Megan Bestwick, Dennis R. Winge

Biochemistry -- Faculty Publications

The assembly of cytochrome c oxidase (CcO) in yeast mitochondria is dependent on a new assembly factor designated Coa2. Coa2 was identified from its ability to suppress the respiratory deficiency of coa1 Δ and shy1 Δ cells. Coa1 and Shy1 function at an early step in maturation of the Cox1 subunit of CcO. Coa2 functions downstream of the Mss51-Coa1 step in Cox1 maturation and likely concurrent with the Shy1-related heme a3 insertion into Cox1. Coa2 interacts with Shy1. Cells lacking Coa2 show a rapid degradation of newly synthesized Cox1. Rapid Cox1 proteolysis also occurs in shy1 Δ cells, suggesting ...


Function And Redox State Of Mitochondrial Localized Cysteine-Rich Proteins Important In The Assembly Of Cytochrome C Oxidase, Oleh Khalimonchuk, Dennis R. Winge Jan 2008

Function And Redox State Of Mitochondrial Localized Cysteine-Rich Proteins Important In The Assembly Of Cytochrome C Oxidase, Oleh Khalimonchuk, Dennis R. Winge

Biochemistry -- Faculty Publications

The cytochrome c oxidase (CcO) complex of the mitochondrial respiratory chain exists within the mitochondrial inner membrane (IM). The biogenesis of the complex is a multi-faceted process requiring multiple assembly factors that function on both faces of the IM. Formation of the two copper centers of CcO occurs within the intermembrane space (IMS) and is dependent on assembly factors with critical cysteinyl thiolates. Two classes of assembly factors exist, one group being soluble IMS proteins and the second class being proteins tethered to the IM. A common motif in the soluble assembly factors is a duplicated Cx9C sequence ...


Pet191 Is A Cytochrome C Oxidase Assembly Factor In Saccharomyces Cerevisiae, Oleh Khalimonchuk, Kevin Rigby, Megan Bestwick, Fabien Pierrel, Paul A. Cobine, Dennis R. Winge Jan 2008

Pet191 Is A Cytochrome C Oxidase Assembly Factor In Saccharomyces Cerevisiae, Oleh Khalimonchuk, Kevin Rigby, Megan Bestwick, Fabien Pierrel, Paul A. Cobine, Dennis R. Winge

Biochemistry -- Faculty Publications

The twin-Cx9C motif protein Pet191 is essential for cytochrome c oxidase maturation. The motif Cys residues are functionally important and appear to be present in disulfide linkages within a large oligomeric complex associated with the mitochondrial inner membrane. The import of Pet191 differs from that of other twin-Cx9C motif class of proteins in being independent of the Mia40 pathway.


Purification From Human Milk Of Matriptase Complexes With Secreted Serpins: Mechanism For Inhibition Of Matriptase Other Than Hai-1, I-Chu Tseng, Feng-Pai Chou, Sheng-Feng Su, Michael Oberst, Nandakumar Madayiputhiya, Ming-Shyue Lee, Jehng-Kang Wang, David E. Sloane, Michael Johnson, Chen-Yong Lin Jan 2008

Purification From Human Milk Of Matriptase Complexes With Secreted Serpins: Mechanism For Inhibition Of Matriptase Other Than Hai-1, I-Chu Tseng, Feng-Pai Chou, Sheng-Feng Su, Michael Oberst, Nandakumar Madayiputhiya, Ming-Shyue Lee, Jehng-Kang Wang, David E. Sloane, Michael Johnson, Chen-Yong Lin

Biochemistry -- Faculty Publications

Matriptase, a type 2 transmembrane serine protease, is predominately expressed by epithelial and carcinoma cells in which hepatocyte growth factor activator inhibitor 1 (HAI-1), a membrane-bound, Kunitz-type serine protease inhibitor, is also expressed. HAI-1 plays dual roles in the regulation of matriptase, as a conventional protease inhibitor and as a factor required for zymogen activation of matriptase. As a consequence, activation of matriptase is immediately followed by HAI-1-mediated inhibition, with the activated matriptase being sequestered into HAI-1 complexes. Matriptase is also expressed by peripheral blood leukocytes, such as monocytes and macrophages; however, in contrast to epithelial cells, monocytes and macrophages ...


Regulated Expression Of Adenoviral Vectors-Based Gene Therapies: Therapeutic Expression Of Toxins And Immune-Modulators, James Curtin, Marianela Candolfi, Mariana Puntel, Weidong Xiong, Akm Ghulam Muhammad, Kurt Kroeger, Sonali Mondkar, Chunyan Liu, Niyati Bondale, Pedro Lowenstein, Maria Castro Jan 2008

Regulated Expression Of Adenoviral Vectors-Based Gene Therapies: Therapeutic Expression Of Toxins And Immune-Modulators, James Curtin, Marianela Candolfi, Mariana Puntel, Weidong Xiong, Akm Ghulam Muhammad, Kurt Kroeger, Sonali Mondkar, Chunyan Liu, Niyati Bondale, Pedro Lowenstein, Maria Castro

Articles

Regulatable promoter systems allow gene expression to be tightly controlled in vivo. This is highly desirable for the development of safe, efficacious adenoviral vectors that can be used to treat human diseases in the clinic. Ideally, regulatable cassettes should have minimal gene expression in the "OFF" state, and expression should quickly reach therapeutic levels in the "ON" state. In addition, the components of regulatable cassettes should be non-toxic at physiological concentrations and should not be immunogenic, especially when treating chronic illness that requires long-lasting gene expression. In this chapter, we will describe in detail protocols to develop and validate first ...


Functional Domains Of The Fatty Acid Transport Proteins: Studies Using Protein Chimeras, Concetta C. Dirusso, Dina Darwis, Thomas Obermeyer, Paul N. Black Jan 2008

Functional Domains Of The Fatty Acid Transport Proteins: Studies Using Protein Chimeras, Concetta C. Dirusso, Dina Darwis, Thomas Obermeyer, Paul N. Black

Biochemistry -- Faculty Publications

Fatty acid transport proteins (FATP) function in fatty acid trafficking pathways, several of which have been shown to participate in the transport of exogenous fatty acids into the cell. Members of this protein family also function as acyl CoA synthetases with specificity towards very long chain fatty acids or bile acids. These proteins have two identifying sequence motifs: The ATP/AMP motif, an approximately 100 amino acid segment required for ATP binding and common to members of the adenylate-forming super family of proteins, and the FATP/VLACS motif that consists of approximately 50 amino acid residues and is restricted to ...


Mapping The Functional Interaction Of Sco1 And Cox2 In Cytochrome Oxidase Biogenesis, Kevin Rigby, Paul A. Cobine, Oleh Khalimonchuk, Dennis R. Winge Jan 2008

Mapping The Functional Interaction Of Sco1 And Cox2 In Cytochrome Oxidase Biogenesis, Kevin Rigby, Paul A. Cobine, Oleh Khalimonchuk, Dennis R. Winge

Biochemistry -- Faculty Publications

Sco1 is implicated in the copper metallation of the CuA site in

Cox2 of cytochrome oxidase. The structure of Sco1 in the metallated

and apo-conformers revealed structural dynamics primarily

in an exposed region designated loop 8. The structural

dynamics of loop 8 in Sco1 suggests it may be an interface for

interactions with Cox17, the Cu(I) donor and/or Cox2. A series

of conserved residues in the sequence motif 217KKYRVYF223 on

the leading edge of this loop are shown presently to be important

for yeast Sco1 function. Cells harboring Y219D, R220D, V221D,

and Y222D mutant Sco1 ...


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 ...