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(1 - 5 of 5)
- Title
- A Quantitative Spectrophotometric Study of Virus Infected KB Cells.
- Creator
- Hilliard, Patricia B., Waddell, Glenn H., Florida Atlantic University
- Abstract/Description
-
The effect of adenovirus and vaccinia infection on KB cells was studied spectrophotometrically by comparing the DNA, RNA, and protein concentrations of infected cells with normal cells. The concentration of these macromolecules in normal cells was established by assaying a large number of KB cultures at different stages of growth. Data were plotted against cell count and time. DNA, RNA, and protein concentrations were found to be linear functions of cell count within a range of 5-25 x 10^5...
Show moreThe effect of adenovirus and vaccinia infection on KB cells was studied spectrophotometrically by comparing the DNA, RNA, and protein concentrations of infected cells with normal cells. The concentration of these macromolecules in normal cells was established by assaying a large number of KB cultures at different stages of growth. Data were plotted against cell count and time. DNA, RNA, and protein concentrations were found to be linear functions of cell count within a range of 5-25 x 10^5 cells/ml. Replicate cultures of KB cells were inoculated with vaccinia or adenovirus at 10 PFU/cell and harvested at 24 hr intervals for 4 days. The cells were counted microscopically at each interval and the concentrations of DNA, RNA, and protein were determined. These results were compared to concentrations and cell counts in uninfected cells and plotted against time. Cell division was inhibited by infection with either of the viruses. At 24 hr post infection (PI) the RNA concentration in infected cells was greater than that found in normal cells. A sharp decrease in RNA content occurred after 24 hr PI until the levels in infected cells were below those in uninfected cells. DNA and protein concentrations were much lower in the infected cells than in the normal cultures. DNA and protein levels increased in the infected cells at 72 hr PI while the DNA concentration in the uninfected cells remained constant. In vaccinia-infected cells the rise in protein synthesis occurred simultaneously with a rise in hemagglutination titer. A similar increase in protein concentration paralleled a rise in complement fixation titer in adenovirus-infected cells.
Show less - Date Issued
- 1970
- PURL
- http://purl.flvc.org/fau/fd/FA00000764, http://purl.flvc.org/fau/fd/FA00000764
- Subject Headings
- Viral antigens, Vaccinia, Adenoviruses
- Format
- Document (PDF)
- Title
- Use of sera from humans and dolphins with lacaziosis and sera from experimentally infected mice forwestern blot analyses of Lacazia loboi antigens.
- Creator
- Mendoza, L.,, Belone, A. F. F., Vilela, R., Rehtanz, Manuela, Bossart, Gregory D., Reif, John S., Fair, Patricia A., Durden, Wendy N., St. Leger, J., Travassos, L. R., Rosa, P. S., Harbor Branch Oceanographic Institute
- Date Issued
- 2008
- PURL
- http://purl.flvc.org/fau/fd/FA00007038
- Subject Headings
- Lobomycosis, Blotting, Western, Antigens, Serum
- Format
- Document (PDF)
- Title
- TUMOR-ASSOCIATED MUC1-TN GLYCOPEPTIDE INTERACTIONS WITH MACROPHAGE GALACTOSE LECTIN.
- Creator
- Beckwith, Donella Marie, Cudic, Mare, Florida Atlantic University, Department of Chemistry and Biochemistry, Charles E. Schmidt College of Science
- Abstract/Description
-
The transformation from normal to malignant phenotype in human cancers is associated with aberrant cell-surface glycosylation. Mucin 1 (MUC1), the heavily glycosylated cell-surface mucin, is altered in both, expression and glycosylation pattern in many cancers. The presence of truncated glycan structures, often capped by sialic acid, commonly known as tumor-associated carbohydrate antigens (TACAs), play key roles in tumor initiations, progression, and metastasis. Accumulating evidence...
Show moreThe transformation from normal to malignant phenotype in human cancers is associated with aberrant cell-surface glycosylation. Mucin 1 (MUC1), the heavily glycosylated cell-surface mucin, is altered in both, expression and glycosylation pattern in many cancers. The presence of truncated glycan structures, often capped by sialic acid, commonly known as tumor-associated carbohydrate antigens (TACAs), play key roles in tumor initiations, progression, and metastasis. Accumulating evidence suggests that expression of TACAs is associated with escape of immune defenses. Human macrophage galactose-type lectin (hMGL, HML, CD301 or CLEC10A), a C-type lectin expressed by antigen presenting cells (APC), is a receptor of mucin-type TACAs, -GalNAc (Thomsen nouvelle antigen; Tn; CD175) and its 2,6-sialylated derivative (sTn; CD175s). To date, the relative contributions of these glycans, as well as underlying peptide backbone, and different degrees of valency, on binding thermodynamics and kinetics with hMGL remains elusive. In order to discern the subtle utility of these distinct features, chemical syntheses of the MUC1, HGVTSAPDTRPAPGSTAPPA tandem repeat sequence, and its site-specific serine (Ser) and threonine (Thr) glycosylated analogs were carried out. Circular dichroism (CD) spectroscopy experiments detected increasing structural order of the Thr glycopeptides compared to its nonglycosylated analogs. Isothermal titration calorimetry (ITC) data analysis of lectin binding to the Thr glycopeptides invariably showed enthalpy-driven processes. Affinity enhancement of the Thr glycopeptides for hMGL occurred relative to free GalNAc, revealing an increasing trend in affinity by one order of magnitude, for mono- (KD = 6-8 μM) to triglycosylated (KD = 600 nM) MUC1 peptides. To delineate the relevance of the solvent structure in the protein carbohydrate recognition process, experiments in D2O were performed, exposing enthalpy-entropy compensation differences. KinITC analysis highlighted prolonged complex lifetimes. Furthermore, atomic force microscopy (AFM) based dynamic single-molecule force spectroscopy (SMFS) provided molecular level insight into the energy landscapes governing recognition of the MUC1(Tn)-hMGL complexes. In summary, our results suggest that contact with hMGL critically depends on the type of TACA, nature of the vicinity surrounding the glycan, and its density. This highlights the importance and current efforts in design of prophylactic and therapeutic cancer vaccines with special emphasis on the synthetic glycopeptide vaccines.
Show less - Date Issued
- 2021
- PURL
- http://purl.flvc.org/fau/fd/FA00013750
- Subject Headings
- Antigens, Tumor-Associated, Carbohydrate, Mucin-1, Cancer vaccines, Glycopeptides
- Format
- Document (PDF)
- Title
- EFFECTS OF SPECIFIC PfEMP1 LIGATION INTERACTIONS WITH ICAM-1, INTEGRIN αVβ3, AND CD36 ON MONOCYTES IN AN IN VITRO MALARIANAÏVE HOST MODEL.
- Creator
- Merritt, Jordan, Oleinikov, Andrew, Florida Atlantic University, Department of Biomedical Science, Charles E. Schmidt College of Science
- Abstract/Description
-
Malaria is a severe global health problem that causes approximately 435,000 deaths per year. Any non-immune individual traveling to malaria endemic regions can be affected too, including humanitarian volunteers, travelers, and US troops. Under physiological conditions, damaged or malaria-infected RBCs would be removed within the spleen, but Plasmodium falciparum infected RBCs (iRBCs) sequester to microvascular endothelial cells to avoid entering the spleen. Adhesion interactions and parasite...
Show moreMalaria is a severe global health problem that causes approximately 435,000 deaths per year. Any non-immune individual traveling to malaria endemic regions can be affected too, including humanitarian volunteers, travelers, and US troops. Under physiological conditions, damaged or malaria-infected RBCs would be removed within the spleen, but Plasmodium falciparum infected RBCs (iRBCs) sequester to microvascular endothelial cells to avoid entering the spleen. Adhesion interactions and parasite sequestration to endothelial cells are mediated by Plasmodium falciparum erythrocyte membrane protein 1 family (PfEMP1) proteins expressed on the iRBC’s surface. The PfEMP1 proteins bind to existing endothelial cell surface receptors that already serve primary functions, including ICAM-1, integrin αVβ3, and CD36. Traditionally, these receptors are explored in the context of endothelial cell sequestration, but this project examines the consequence of receptor::PfEMP1 interaction on immune cells, namely monocyte-like THP-1 cells.
Show less - Date Issued
- 2019
- PURL
- http://purl.flvc.org/fau/fd/FA00013398
- Subject Headings
- Malaria, Plasmodium falciparum, Integrins, Monocytes, Intercellular Adhesion Molecule-1, CD36 Antigens, Adhesiveness
- Format
- Document (PDF)
- Title
- Systemic Lupus Erythematosus: A.Studies on an ldiotype Specific Dendritic Cell Vaccine. B.Association of Lupus Calcinosis with Calcifying Nanoparticles.
- Creator
- Keating, Patricia, Florida Atlantic University, Hartmann, James X., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Systemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier...
Show moreSystemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier proteins, conjugated to lgG, was evaluated, and KLH found to be the most efficient for antigen uptake. The cognate dendritic cells were evaluated for their capacity to activate autologous T cells, and generate a Th1 mediated response which was evaluated by proliferation assays and interferony secretion. During the vaccine study a patient presented with panniculitis, CREST syndrome and a calcinotic exudate. A secondary goal of my study was to analyze this exudate. Calcifying nanoparticles were isolated in lymphocyte culture medium. They were characterized by Von Kassa staining for hydroxyapatite, solubilization by the calcium chelating agent EDT A, and light and scanning electron microscopy. A novel method was developed, using a specific monoclonal antibody, to analyze the calcifying nanoparticles. This method allowed for an approximate quantification of the particles. These particles increased in numbers when incubated for different time periods, and their numbers were decreased when incubated in the presence of minocyclin. Concomitantly, the panniculi in the patient underwent remission with long term antibiotic therapy. CNPs were also obtained from fetal bovine serum and human plasma samples from both lupus patients and healthy donors. Peripheral blood mononuclear cells from healthy donors and lupus patients were analyzed in vitro for their reactivity when incubated in the presence of a biofilm, generated by the calcifying nanoparticles. Viability, proliferation, and co-stimulatory marker up-regulation were determined in the presence or absence of the particles. Osteopontin was found highly expressed in the supernatants of the cells grown with CNPs. Microarray of the mononuclear cells of a healthy donor and a lupus patient incubated in the presence or absence of CNPs was performed, and the results coincided with those determined in vitro.
Show less - Date Issued
- 2007
- PURL
- http://purl.flvc.org/fau/fd/FA00000865
- Subject Headings
- Systemic lupus erythematosus--Molecular aspects, Systemic lupus erythematosus--Immunological aspects, Cell surface antigens, Autoimmune diseases--Research, Monoclonal antibodies--Diagnostic use
- Format
- Document (PDF)
