Current Search: HIV-1 (x)
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Title
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HIV-1 INFECTION MODULATES CD4+ T CELL GENE EXPRESSION TO INDUCE A QUIESCENT PHENOTYPE.
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Creator
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Mauer, Christopher, Caputi, Massimo, Florida Atlantic University, Department of Biomedical Science, Charles E. Schmidt College of Medicine
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Abstract/Description
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The largest barrier to treatment of HIV-1 infection is the establishment of a viral reservoir constituted mostly by quiescent latently infected CD4+ T cells. This reservoir is formed through two processes: i) the infection of resting CD4+ T cells; both naïve and memory, ii) the infection of activated CD4+ T cells which then become quiescent infected cells. One goal of this project was to understand the gene expression changes occurring in naïve CD4+ T cells following activation and subsequent...
Show moreThe largest barrier to treatment of HIV-1 infection is the establishment of a viral reservoir constituted mostly by quiescent latently infected CD4+ T cells. This reservoir is formed through two processes: i) the infection of resting CD4+ T cells; both naïve and memory, ii) the infection of activated CD4+ T cells which then become quiescent infected cells. One goal of this project was to understand the gene expression changes occurring in naïve CD4+ T cells following activation and subsequent HIV-1 infection and how this may contribute to the establishment of a latent infection in these cells. Utilizing RNA-Seq and a series of validation assays we have identified several genes which are regulated in opposite directions during activation versus infection which we termed DEOC genes. The DEOC genes include a group of physically- and functionally-associated proteins which are key regulators of T cell activation, the cell cycle, cellular proliferation, and cellular quiescence, suggesting that modulation of these DEOC genes may help transition the infected-activated cell from an activated state to a quiescent/resting state to induce a latent infection.
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Date Issued
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2022
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PURL
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http://purl.flvc.org/fau/fd/FA00014033
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Subject Headings
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HIV-1, Virus Latency
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Format
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Document (PDF)
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Title
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DEVELOPING PLASMA-BASED DIAGNOSTICS: FROM PANCREATIC DUCTAL ADENOCARCINOMA TO HIV.
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Creator
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Makler, Amy, Asghar, Waseem, Florida Atlantic University, Department of Biological Sciences
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Abstract/Description
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Plasma-based diagnostics are ideal for detecting a variety of diseases because they offer a method of detection that is minimally invasive, readily available, and easy to use for monitoring patients as they progress through a disease or respond to treatment. The only serum marker for PDAC is CA19-9 which lacks specificity, has limited sensitivity, and is unreliable for early detection. It is therefore of great importance to develop a diagnostic that is viable for screening and early detection...
Show morePlasma-based diagnostics are ideal for detecting a variety of diseases because they offer a method of detection that is minimally invasive, readily available, and easy to use for monitoring patients as they progress through a disease or respond to treatment. The only serum marker for PDAC is CA19-9 which lacks specificity, has limited sensitivity, and is unreliable for early detection. It is therefore of great importance to develop a diagnostic that is viable for screening and early detection. Exosomal miRNA were determined via bioinformatics analyses and then examined in PDAC cell lines to identify markers with greatest potential. These markers were then examined in plasma from PDAC patients and control groups. Four markers, miR-93-5p, miR-339-3p, miR-425-5p, and miR-425-3p, emerged as the most viable biomarker panel with the ability to detect PDAC in 100% of the early stages (N=5) compared to CA19-9 which showed increased levels in only one patient with early stage PDAC. Additionally, the diagnostic has a specificity of 80% and a sensitivity of 94.7%, making it comparable to CA19-9, and may even be beneficial for use in conjunction with CA19-9. A plasma-based diagnostic was also developed for multi-strain HIV-1 detection utilizing the loop-mediated isothermal amplification (LAMP) method. LAMP primers were developed against the integrase and vpr regions of the HIV-1 genome. They were tested first in cultured HIV samples and then examined for their ability to amplify HIV-1 subtypes A-G. The integrase primer set provided a reliable means of diagnosing all 55 strains and isolates in under 30 minutes, whereas vpr was inconsistent and exhibited high variability in detecting the HIV subtypes. Our limit of detection for B-subtype with integrase was 30 viral copies/reaction. This could provide the basis for a novel, point-of-care diagnostic for use in underdeveloped regions.
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Date Issued
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2023
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PURL
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http://purl.flvc.org/fau/fd/FA00014141
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Subject Headings
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Plasma, Biomarkers, Carcinoma, Pancreatic Ductal, HIV-1, Diagnosis
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Format
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Document (PDF)
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Title
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DEVELOPMENT OF POINT-OF-CARE ASSAYS FOR DISEASE DIAGNOSTIC AND TREATMENT MONITORING FOR RESOURCE CONSTRAINED SETTINGS.
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Creator
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Sher, Mazhar, Asghar, Waseem, Florida Atlantic University, Department of Computer and Electrical Engineering and Computer Science, College of Engineering and Computer Science
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Abstract/Description
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This thesis aims to address the challenges of the development of cost-effective and rapid assays for the accurate counting of CD4+ T cells and quantification of HIV-1 viral load for resource-constrained settings. The lack of such assays has severely affected people living in disease prevalent areas. CD4+ T cells count information plays a vital role in the effective management of HIV-1 disease. Here, we present a flow-free magnetic actuation platform that uses antibody-coated magnetic beads to...
Show moreThis thesis aims to address the challenges of the development of cost-effective and rapid assays for the accurate counting of CD4+ T cells and quantification of HIV-1 viral load for resource-constrained settings. The lack of such assays has severely affected people living in disease prevalent areas. CD4+ T cells count information plays a vital role in the effective management of HIV-1 disease. Here, we present a flow-free magnetic actuation platform that uses antibody-coated magnetic beads to efficiently capture CD4+ T cells from a 30 μL drop of whole blood. On-chip cell lysate electrical impedance spectroscopy has been utilized to quantify the isolated CD4 cells. The developed assay has a limit of detection of 25 cells per μL and provides accurate CD4 counts in the range of 25–800 cells per μL. The whole immunoassay along with the enumeration process is very rapid and provides CD4 quantification results within 5 min time frame. The assay does not require off-chip sample preparation steps and minimizes human involvement to a greater extent. The developed impedance-based immunoassay has the potential to significantly improve the CD4 enumeration process especially for POC settings.
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Date Issued
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2020
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PURL
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http://purl.flvc.org/fau/fd/FA00013495
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Subject Headings
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Point-of-care testing, Diagnostic tests, Immunoassay, HIV-1, Microfluidic devices
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Format
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Document (PDF)