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You searched for +publisher:"Temple University" +contributor:("Suhadolnik, Robert J."). Showing records 1 – 3 of 3 total matches.

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Temple University

1. Teets, Bryan Wilson. SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT.

Degree: PhD, 2011, Temple University

Biochemistry

Retinoic acid (RA) is critical for embryonic development and cell differentiation. Previous work in our laboratory has shown that blocking the RA-dependent increase in Pre-â cell leukemia transcription factors (PBX) mRNA and protein levels in P19 cells prevents them from differentiating to either endodermal or neuronal cells. This suggests that PBX is an important regulator of RA-induced differentiation of P19 cells. A microarray analysis was performed to identify PBX regulated genes, utilizing the empty vector P19 (TO3) and antisense to PBX (AS2) cell lines, during RA-induced differentiation of P19 cells into endodermal or neuronal cells. Among the genes identified by the microarray, Dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1 (DAX-1) and steroidogenic factor 1 (SF-1) were identified to be directly or indirectly regulated by PBX. Both DAX-1 and SF-1 proteins have only recently been reported to be present in preimplantation mouse embryos prior to the expression of steroidogenic enzymes, suggesting they may play a role in early mouse embryogenesis. To determine the roles of DAX-1 and SF-1 during RA-dependent differentiation, P19 cells that inducibly express either FLAG-DAX-1 or FLAG-SF-1 upon removal of doxicyclin were prepared. We found that overexpression of FLAG-DAX-1 had no effect on the RA-induced differentiation of P19 cells. However, FLAG-SF-1 overexpression prevented the RA-dependent loss of Oct-4, DAX-1 and the increase in COUP-TFI, COUP-TFII, and Ets-1 mRNA levels during the commitment stages of both endodermal and neuronal differentiation. Surprisingly, continued expression of SF-1 for seven days caused a RA-independent loss of Oct-4 protein. However, cells which continued to express SF-1 for seven days did not terminally differentiate into endodermal or neuronal cells in response to RA treatment. In addition, we found evidence for a feedback loop, where PBX reduces SF-1 mRNA expression and continued SF-1 expression blocks the RA-dependent increase in PBX protein levels. Our findings suggest that SF-1 plays a novel role in P19 cells where its level of expression is critical for the differentiation state of the cells. At basal levels SF-1 maintains the pluripotent state of the cells, while SF-1 levels must be dramatically reduced for cells to differentiate into both endodermal and neuronal cells upon RA treatment. However, at elevated levels above basal, SF-1 inhibits Oct-4 expression and leads to the induction of the expression of steroidogenic enzymes with a pattern consistent with adrenal cells in a RA-independent fashion. Taken together these data suggest that SF-1 plays a much more dynamic role in P19 cells than previously reported.

Temple University – Theses

Advisors/Committee Members: Soprano, Dianne R., Gamero, Ana, Stitt, Barbara L., Suhadolnik, Robert J..

Subjects/Keywords: Biochemistry; adrenal; Differentiation; neuronal; P19; PBX; Vitamin A

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APA (6th Edition):

Teets, B. W. (2011). SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT. (Doctoral Dissertation). Temple University. Retrieved from http://digital.library.temple.edu/u?/p245801coll10,124708

Chicago Manual of Style (16th Edition):

Teets, Bryan Wilson. “SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT.” 2011. Doctoral Dissertation, Temple University. Accessed September 24, 2020. http://digital.library.temple.edu/u?/p245801coll10,124708.

MLA Handbook (7th Edition):

Teets, Bryan Wilson. “SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT.” 2011. Web. 24 Sep 2020.

Vancouver:

Teets BW. SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT. [Internet] [Doctoral dissertation]. Temple University; 2011. [cited 2020 Sep 24]. Available from: http://digital.library.temple.edu/u?/p245801coll10,124708.

Council of Science Editors:

Teets BW. SF-1, BUT NOT DAX-1, PREVENTS P19 CELLS FROM DIFFERENTIATING TO EITHER TROMA-1 OR TUJ1 POSITIVE CELLS UPON RA-TREATMENT. [Doctoral Dissertation]. Temple University; 2011. Available from: http://digital.library.temple.edu/u?/p245801coll10,124708


Temple University

2. Roberts, Sean Anthony. A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS.

Degree: PhD, 2010, Temple University

Microbiology and Immunology

Since it emerged as an infectious agent in 1981, the human immunodeficiency virus type 1 (HIV-1) is continually disseminated and remain fatal to the majority of those infected. Strategies including highly active retroviral therapies (HAART) with nucleoside analogues and protease inhibitors have shown limited success in therapy due to the virus' ability to evolve rapidly at every replication cycle as a consequence of it's highly error prone reverse transcriptase, generating resistant retroviral strains and in addition to latent HIV-1 reservoirs. Thirty years of research efforts to find a cure or to generate a vaccine has been met with failure. It is, therefore, of necessity to broaden our paradigm of therapy for the treatment and eventual cure of HIV-1 infection. In this study, I look beyond the current anti-retroviral strategies and instead rely on the mammalian host immune system to inhibit HIV-1 replication through molecular genetic manipulation. Here, we approach the inhibition of HIV-1 replication by up-regulation of the innate antiviral pathway that is natural to mammalian cells. HIV-1 derived self-inactivating lentiviral (SIN) vectors were designed and constructed to deliver the antiviral payloads of two antiviral enzymes, p68 kinase (PKR) and 2'-5' oligoadenlyate synthetase (2-5OAS), to target cell, SupT1 lymphoblastoid cells and CD4+ T lymphocytes under the control of a constitutive cytomegalovirus (CMV) promoter. These data here demonstrates a significant inhibition of HIV-1 replication in cells transduced with the anti HIV-1 transgenes PKR and 2-5OAS as determined by HIV-1 induced syncytia formation and HIV-1 p24 antigen capture assay. Furthermore, here demonstrated is an increase up-regulation of PKR and 2-5OAS 96 hr post cell transduction in all the clones when compared to pHIV empty vector control. These results demonstrate that the over-expression of PKR and 2-5OAS can inhibit HIV-1 replication and also confirm the involvement of PKR and 2-5OAS in the IFN-associated antiviral pathway against HIV-1 infection.

Temple University – Theses

Advisors/Committee Members: Henderson, Earl E., Suhadolnik, Robert J., Ganea, Doina, Tsygankov, Alexander, De Riel, Jon K..

Subjects/Keywords: Biology, Microbiology; Biology, virology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Roberts, S. A. (2010). A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS. (Doctoral Dissertation). Temple University. Retrieved from http://digital.library.temple.edu/u?/p245801coll10,99935

Chicago Manual of Style (16th Edition):

Roberts, Sean Anthony. “A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS.” 2010. Doctoral Dissertation, Temple University. Accessed September 24, 2020. http://digital.library.temple.edu/u?/p245801coll10,99935.

MLA Handbook (7th Edition):

Roberts, Sean Anthony. “A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS.” 2010. Web. 24 Sep 2020.

Vancouver:

Roberts SA. A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS. [Internet] [Doctoral dissertation]. Temple University; 2010. [cited 2020 Sep 24]. Available from: http://digital.library.temple.edu/u?/p245801coll10,99935.

Council of Science Editors:

Roberts SA. A GENE THERAPY APPROACH TO THE INHIBITION OF HIV-1 REPLICATION BY RESTORATION OF INNATE ANTIVIRAL DEFENSE PATHWAYS. [Doctoral Dissertation]. Temple University; 2010. Available from: http://digital.library.temple.edu/u?/p245801coll10,99935


Temple University

3. Shikov, Sergei. Structural Determinants for Heparin Binding in Human Coagulation Factor XI.

Degree: PhD, 2008, Temple University

Biochemistry

Coagulation factor XI plays an important role in the consolidation phase of blood coagulation. Previous studies from our laboratory and others have demonstrated that zymogen factor XI (FXI) binds to heparin with moderate (KD~110 nM) affinity via residues (K252, K253 and K255) located in the apple 3 (A3) domain of the molecule. In contrast, the enzyme, factor XIa (FXIa), was shown to bind to heparin with significantly higher affinity (~1.5 nM by ELISA) via residues (K529, R530 and R532) within the catalytic domain (CD). The interaction between heparin and FXIa potentiates the inhibition of FXIa by protease nexin-2 by 10-fold. In addition, related polyanions heparin and dextran sulfate inhibit the catalytic activity of FXIa. The present study was designed to determine the relative contributions of positively charged residues as well as the dimeric structure of FXI in heparin binding. During this project, wtFXI, FXIR504A, FXIK505A, FXIR507A, FXIR529A, FXIR530A, FXIR532A, and FXIR586A have been expressed and purified. All mutants were homogenous and identical to wtFXI on SDS-PAGE, clotting assays and 1G5 monoclonal antibody binding studied by SPR. In addition, monomeric FXI C321S/K331A was expressed and purified. Utilizing an ELISA assay, no difference in the affinity for heparin between FXIa and FXI was found. Surface plasmon resonance (SPR) data collected for FXI clearly indicate a complex interaction which does not conform to a simple 1:1 Langmuir binding model making it difficult to obtain quantitative information. The complexity of FXI interactions with heparin is likely to arise from the multivalent nature of the binding, in which both protein and heparin have multiple binding sites. Two positively charged residues in the FXI catalytic domain, FXIR507A and FXIR532A, were found to be particularly important for interaction with heparin. The FXIR507A and FXIR532A mutants demonstrated ~ 65% and ~50% decreases respectively in total number of heparin binding sites based on ELISA. Also, the apparent dissociation constants for FXIR507A (KDapp ~13 nM) and FXIR532A (KDapp ~21 nM ) were 6 and 10-fold increased respectively compared with 2.1 nM for the wtFXI. Mutant FXIR586A also demonstrated a defect in affinity (KDapp ~ 13 nM) without an effect on the Bmax. The monomeric FXIC321S/R331A was also characterized for its ability to bind heparin compared with wtFXI. Surprisingly, the monomeric FXI displayed defective binding to heparin according to ELISA (KDapp ~ 30 nM) and SPR methods. Thus, the unique homodimeric structure of FXI in addition to the residues both in its catalytic and A3 domain chains are necessary for high-affinity heparin binding.

Temple University – Theses

Advisors/Committee Members: Walsh, Peter N., Shore, Scott K., Collins, Jimmy H., Suhadolnik, Robert J., Soslau, Gerald.

Subjects/Keywords: Chemistry, Biochemistry; Biophysics, General; Heparin; blood coagulation; ELISA; SPR

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Shikov, S. (2008). Structural Determinants for Heparin Binding in Human Coagulation Factor XI. (Doctoral Dissertation). Temple University. Retrieved from http://digital.library.temple.edu/u?/p245801coll10,13460

Chicago Manual of Style (16th Edition):

Shikov, Sergei. “Structural Determinants for Heparin Binding in Human Coagulation Factor XI.” 2008. Doctoral Dissertation, Temple University. Accessed September 24, 2020. http://digital.library.temple.edu/u?/p245801coll10,13460.

MLA Handbook (7th Edition):

Shikov, Sergei. “Structural Determinants for Heparin Binding in Human Coagulation Factor XI.” 2008. Web. 24 Sep 2020.

Vancouver:

Shikov S. Structural Determinants for Heparin Binding in Human Coagulation Factor XI. [Internet] [Doctoral dissertation]. Temple University; 2008. [cited 2020 Sep 24]. Available from: http://digital.library.temple.edu/u?/p245801coll10,13460.

Council of Science Editors:

Shikov S. Structural Determinants for Heparin Binding in Human Coagulation Factor XI. [Doctoral Dissertation]. Temple University; 2008. Available from: http://digital.library.temple.edu/u?/p245801coll10,13460

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