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Development and Optimization of AlphaScreen Assay for Discovery of Galectin-1/-3 Inhibitors
Title
Development and Optimization of AlphaScreen Assay for Discovery of Galectin-1/-3 Inhibitors.
Creator
Rivero, Yaima, Fitzgerald, Forrest, Cudic, Mare
Abstract/Description
Galectin-1 is a beta-galactoside-binding protein implicated in regulating apoptosis, cell proliferation and cell differentiation. The objective of our research was to develop and optimize an assay format for the discovery of new inhibitors of galectin-1 using AlphaScreen technology in a competitive binding configuration. Our efforts were hampered by the weak binding affinities of galectin-1 for its binding ligands (μM range). Consequently, the AlphaScreen assay could not have been developed...
Show moreGalectin-1 is a beta-galactoside-binding protein implicated in regulating apoptosis, cell proliferation and cell differentiation. The objective of our research was to develop and optimize an assay format for the discovery of new inhibitors of galectin-1 using AlphaScreen technology in a competitive binding configuration. Our efforts were hampered by the weak binding affinities of galectin-1 for its binding ligands (μM range). Consequently, the AlphaScreen assay could not have been developed at the level that would satisfy the guidelines from the National Chemical Genomics Center. Nevertheless, we have optimized the AlphaScreen assay for galectin-3, another member of galectin family, and screened an FDA approved oncology drug library (n = 101) from the NIH. We identified three possible inhibitors of galectin-3. These compounds showed a positive activity in dose response experiments: bleomycin (IC50 = 25.7 nM), cisplatin (IC50 = 127 nM), and mitoxantrone HCl (IC50 = 662 nM).
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Date Issued
2018
PURL
http://purl.flvc.org/fau/fd/FAU_SR00000050
Subject Headings
College students --Research --United States.
Format
Document (PDF)
Roles of adjuvant and route of vaccination in antibody response and protection engendered by a synthetic matrix protein 2-based influenza A virus vaccine in the mouse
Title
Roles of adjuvant and route of vaccination in antibody response and protection engendered by a synthetic matrix protein 2-based influenza A virus vaccine in the mouse.
Creator
Mozdzanowska, Krystyna, Zharikova, Darya, Cudic, Mare, Otvos, Laszlo, Gerhard, Walter
Date Issued
2007-10-27
PURL
http://purl.flvc.org/fcla/dt/3327982
Format
Document (PDF)
Characterization of Disulfide Constrained Natural Peptides
Title
Characterization of Disulfide Constrained Natural Peptides.
Creator
Hoggard, Mickelene F., Cudic, Mare, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Chemistry and Biochemistry
Abstract/Description
The use of peptide drugs has gained popularity recently. Peptides are attractive drug targets due to their high specificity and potency towards their biological targets. A drawback for peptide drugs is a lack of stability for oral delivery. Two classes of disulfide-rich peptides, conotoxins and cyclotides, have been shown to have higher stability than linear peptides thanks to their disulfide connectivity. Conotoxins are present in the venom of cone snails, a carnivorous marine mollusk that...
Show moreThe use of peptide drugs has gained popularity recently. Peptides are attractive drug targets due to their high specificity and potency towards their biological targets. A drawback for peptide drugs is a lack of stability for oral delivery. Two classes of disulfide-rich peptides, conotoxins and cyclotides, have been shown to have higher stability than linear peptides thanks to their disulfide connectivity. Conotoxins are present in the venom of cone snails, a carnivorous marine mollusk that preys upon fish, worms, or other mollusks. Conotoxins are promising drugs leads with great prospects in the treatment of diseases and disorders such as chronic pain, multiple sclerosis and Parkinson’s and Alzheimer’s diseases. Cyclotides, which are cyclic cysteine knot containing peptides, isolated from the Violaceae (violet), Rubiaceae (coffee), and Cucurbitaceae (cucurbit) families and they have a wide range of biological activities, such as anti-HIV, uterotonic, and antimicrobial. P-superfamily framework IX conotoxins (C-C- C-CXC- C) contain the same cysteine framework, homologous sequences, and similar 3D structures to cyclotides. The knot containing conotoxins have been identified in several Conus species, but this work focuses on those from Conus brunneus, Conus purpurascens, and Conus gloriamaris. The cysteine knot motif of cyclotides and P-superfamily conotoxins is characterized by a cyclic backbone and six-conserved cysteine residues that form the three-disulfide bridges of the “knot”. This motif provides cyclotides and conotoxins with superior stability against thermal, chemical, and enzymatic degradation; marking them as potential frameworks for peptide drug delivery. Presented are details on the isolation of conotoxins and cyclotides, from Viola tricolor, and the characterization of their activity in the well-characterized Drosophila melanogaster giant fiber system (GFS) neuronal circuit, which contains GAP, acetylcholine, and glutamate synapses. The transcriptomes of two Conus brunneus specimens were assembled and mined for P-superfamily framework IX conotoxins. Eleven mature P-superfamily framework IX conotoxins were identified in the crude venom.
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Date Issued
2018
PURL
http://purl.flvc.org/fau/fd/FA00005955
Subject Headings
Peptide drugs, Cyclotides, Conotoxins
Format
Document (PDF)
Antibodies elicited by the first non-viral prophylactic cancer vaccine show tumor-specificity and immunotherapeutic potential
Title
Antibodies elicited by the first non-viral prophylactic cancer vaccine show tumor-specificity and immunotherapeutic potential.
Creator
Lohmueller, Jason J., Sato, Shuji, Popova, Lana, Chu, Isabel M., Tucker, Meghan A., Barberena, Roberto, Innocenti, Gregory M., Cudic, Mare, Ham, James D., Cheung, Wan Cheung, Polakiewicz, Roberto D., Finn, Olivera J.
Abstract/Description
MUC1 is a shared tumor antigen expressed on >80% of human cancers. We completed the first prophylactic cancer vaccine clinical trial based on a non-viral antigen, MUC1, in healthy individuals at-risk for colon cancer. This trial provided a unique source of potentially effective and safe immunotherapeutic drugs, fully-human antibodies affinity-matured in a healthy host to a tumor antigen. We purified, cloned, and characterized 13 IgGs specific for several tumor-associated MUC1 epitopes with a...
Show moreMUC1 is a shared tumor antigen expressed on >80% of human cancers. We completed the first prophylactic cancer vaccine clinical trial based on a non-viral antigen, MUC1, in healthy individuals at-risk for colon cancer. This trial provided a unique source of potentially effective and safe immunotherapeutic drugs, fully-human antibodies affinity-matured in a healthy host to a tumor antigen. We purified, cloned, and characterized 13 IgGs specific for several tumor-associated MUC1 epitopes with a wide range of binding affinities. These antibodies bind hypoglycosylated MUC1 on human cancer cell lines and tumor tissues but show no reactivity against fully-glycosylated MUC1 on normal cells and tissues. We found that several antibodies activate complement-mediated cytotoxicity and that T cells carrying chimeric antigen receptors with the antibody variable regions kill MUC1+ target cells, express activation markers, and produce interferon gamma. Fully-human and tumor-specific, these antibodies are candidates for further testing and development as immunotherapeutic drugs.
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Date Issued
2016-10-22
PURL
http://purl.flvc.org/fau/fd/FAUIR000012
Format
Citation