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A morphological, histochemical and experimental study of the prostate gland and seminal vesicles of the guinea pig, with special referenceto the stroma陳良, Chan Leung, Franky. January 1989 (has links)
published_or_final_version / abstract / toc / Anatomy / Doctoral / Doctor of Philosophy
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Adrenergic control and its mechanism of stimulation of electrogenic anion secretion in primary cultures of rat epididymal eipthelialcells陳浦棠, Chan, Po-tong, Timothy. January 1990 (has links)
published_or_final_version / Physiology / Master / Master of Philosophy
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Bioartificial matrices to modulate epithelial morphogenesisEnemchukwu, Nduka Obichukwu 12 January 2015 (has links)
Acute injury of major epithelial organ systems (kidney, liver, lung, etc.) is collectively a principal cause of death worldwide. Regenerative medicine promises to meet these human health challenges by harnessing intrinsic cellular processes to repair or replace damaged tissues.
Epithelial morphogenesis is a hard-wired, multicellular differentiation program that dynamically integrates microenvironmental cues to coordinate cell fate processes including adhesion, migration, proliferation, and polarization. Thus, epithelial morphogenesis is an instructive mode of tissue assembly, maintenance, and repair. Three-dimensional epithelial cell cultures in natural basement membrane (BM) extracts produce hollow, spherical cyst structures and have indicated that the BM provides the critical cell adhesion ligands to facilitate cell survival, stimulate proliferation, and promote polarization and lumen formation. However, the utility of natural BMs for detailed studies is generally limited by lot-to-lot variations, uncontrolled cell adhesive interactions, or growth factor contamination.
The goal of this thesis was to engineer bioartificial extracellular matrices (ECM) that would support and modulate epithelial cyst morphogenesis. We have engineered hydrogels, based on a multi-arm maleimide-terminated poly (ethylene glycol) (PEG-4MAL), that present cell adhesive molecules and enzymatic degradation substrates and promote polarized epithelial cyst differentiation in vitro.
To investigate the influence of matrix physical and biochemical signals on cyst morphogenesis, we independently varied the polymer weight percentage (wt%), the density of a cell adhesion ligand (RGD), and crosslink degradation rates of the hydrogels. Then, we evaluated functional outcomes including Madin-Darby canine kidney (MDCK II) epithelial cell survival, proliferation, cyst polarization, and lumen formation. We found that cell proliferation, but not cell survival, was sensitive to the polymer wt%, which is related to elastic modulus and crosslink density. This result defined a working range of PEG-4MAL concentration (3.5% - 4.5%) that promotes robust proliferation. Analysis of mature cysts indicated that 4.0% and 4.5% gels produced cysts resembling those typically grown in type I collagen gels while 3.5% gels produced cysts with higher incidence of inverted polarity and multiple lumens. Perturbation of matrix degradability using a slow-degrading crosslink peptide or matrix metalloproteinase inhibitors showed that the rate of matrix degradation exerts major influence on cyst growth in PEG-4MAL gels. We employed 4.0% PEG-4MAL hydrogels with RGD ligand density ranging over 0 – 2000 uM to discover that (1) lumen formation was eliminated in the absence of RGD, (2) extent of lumen formation increased with increasing RGD concentration, and (3) cyst polarity was inverted below a threshold of integrin binding to RGD.
Together, these results show that the biochemical and physical properties of the matrix, particularly integrin binding and matrix degradability, effectively modulate establishment of apico-basal polarity and lumen phenotypes in MDCK II epithelial cyst structures. Furthermore, these studies validate PEG-4MAL hydrogels as a powerful culture platform to enable detailed investigation of matrix-directed modulation of epithelial morphogenesis.
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Identification, isolation and characterization of proinsulin producing thymic cellsPalumbo, Michael O. January 2007 (has links)
The finding that more than 152 tissue-restricted antigens are expressed by thymic medullary epithelial cells is redefining the importance of thymic central tolerance induction in the prevention of autoimmune diseases. One of the tissue-restricted antigens in the thymus is proinsulin, and in both mice and humans, reduced thymic proinsulin levels have been shown to predispose to Type 1 diabetes. Using transgenic mice expressing a functional beta-Galactosidase gene under the regulation of the Ins2 promoter we have determined that between 1-3% of all medullary thymic epithelial cells express proinsulin and that these cells are frequently part of the Hassall's Corpuscles like structures in mice. Using a cross between the beta-Galactosidase expressing mice and Immortomice (expressing SV40 large T Antigen under the regulation of the MHC I promoter), we have isolated and cultured two proinsulin and two non-proinsulin producing medullary epithelial cell lines. Microarray analysis and RT-PCR analysis of the cell lines revealed the over-expression of approximately 50 genes (>4 fold or more) in the proinsulin producing lineage, versus the non proinsulin producing lineage, and approximately half the over-expressed genes can be considered tissue-restricted antigens. We do not find any evidence for chromosomal clustering of the over-expressed genes nor do we report the expression of any other pancreatic n-cell antigens or specific pancreatic proinsulin regulatory proteins (Pdx-1, Glut-2 or GCK) within the proinsulin producing cell lines but we do detect their expression in whole thymus. Our results suggest that chromosomal clustering is not a phenomenon associated with thymic tissue-restricted antigen expression and that the mechanisms allowing for thymic tissue-restricted antigen expression are not related to the expression mechanisms of such antigens in peripheral tissues.
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Epithelial cells: an immune modulator in the context of inflammatory bowel diseasesBacker, Jody Lynn Unknown Date
No description available.
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Interaction between Mycobacterium tuberculosis and pulmonary epithelium.Ashiru, Olubisi T. January 2013 (has links)
Background
Mycobacterium tuberculosis isolates such as the Beijing and F15/LAM4/KZN families
dominate in patients. The emergence of extensively drug resistant (XDR) M. tuberculosis
isolates raises concern. The need to better understand the pathogenesis of M. tuberculosis
isolates resulted in this work.
Methods
M. tuberculosis clinical isolates that belonged to the Beijing and F15/LAM4/KZN
families, isolates with unique DNA fingerprints and laboratory strains were used. Isolates
were grown in the presence of oxygen and then exposed to A549 alveolar and BBM
bronchial epithelial cells. The number of bacilli that adhered to the epithelial cells were
viewed and counted using light microscopy. Isolates grown in the presence of oxygen and
under oxygen deprivation were used for subsequent assays. Invasion of A549 and BBM
cells by isolates grown under these different circumstances was investigated. Based on
the results, the remaining assays were performed with A549 cells only. Cytotoxicity was
quantified using the Cyto Tox96 Non-Radioactive Cytotoxicity Assay kit.
Morphological changes in A549 cells after exposure to the isolates were observed using
the scanning electron microscopy (SEM). Real-time quantitative PCR was performed to
assess the relative expression levels of four genes potentially associated with virulence
(hbhA; mdp1; fdxA; hspX). Results were normalized against 16S rRNA and ftsZ gene
transcription and reported as fold difference as compared to H37Rv.
Results
All isolates adhered to and invaded A549 cells in significantly higher numbers than BBM
cells (P<0.0029). Isolates grown under oxygen deprivation displayed higher levels of
virulence than their aerobic phenotype. Grouped together, the isolates belonging to the
Beijing and F15/LAM4/KZN families of strains showed greater adhesion capacity (28%)
than isolates with unique DNA fingerprints (5%) (P<0.05%). Three F15/LAM4/KZN
isolates (two XDR-variants), were at least twice as invasive (>33%) as the most invasive
Beijing isolate (15%) (P<0.05). The highest cytotoxicity level (35.7%) was produced by
an XDR-F15/LAM4/KZN strain. SEM revealed bleb-like structures on bacterial cells
grown under oxygen deprivation. Beijing and XDR-F15/LAM4/KZN isolates had the
highest number of projections (16+5 per bacillus. The expression levels of all four genes
were highest in Beijing and F15/LAM4/KZN isolates grown under oxygen deprivation
and exposed to A549 cells.
Conclusions
Beijing and F15/LAM4/KZN strains are more virulent and their successful spread might
be related to their interaction with alveolar epithelium. M. tuberculosis pathogenesis
studies should include isolates grown under oxygen deprivation. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2013.
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Anillin, An Organizer of CytokinesisHeshmati, Fatemeh 15 November 2013 (has links)
Anillin is a highly conserved multi-domain cytoskeletal protein that provides a spatial and temporal scaffold for contractile ring proteins to ensure successful cytokinesis. We have looked at the temporal order of anillin and septin recruitment to the cleavage furrow using time-lapse microscopy and found that anillin localizes to the furrow in early anaphase while septins appear there later in an anillin-dependent manner. We also characterized the effect of anillin depletion in different cell lines and observed that septins and myosin delocalize in the absence of anillin in Tet-ON HeLa, AD293 and ARPE-19 cells but not in wild type HeLa cells. Asymmetric furrow formation was also investigated using the epithelial cell model: MDCK cells. Depletion of anillin and SEPT9 in MDCK cells was achieved using lentivirus shRNA constructs and this revealed that anillin or SEPT9 depletion did not affect asymmetric cytokinesis, although localization of SEPT 9 was affected by anillin depletion.
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Anillin, An Organizer of CytokinesisHeshmati, Fatemeh 15 November 2013 (has links)
Anillin is a highly conserved multi-domain cytoskeletal protein that provides a spatial and temporal scaffold for contractile ring proteins to ensure successful cytokinesis. We have looked at the temporal order of anillin and septin recruitment to the cleavage furrow using time-lapse microscopy and found that anillin localizes to the furrow in early anaphase while septins appear there later in an anillin-dependent manner. We also characterized the effect of anillin depletion in different cell lines and observed that septins and myosin delocalize in the absence of anillin in Tet-ON HeLa, AD293 and ARPE-19 cells but not in wild type HeLa cells. Asymmetric furrow formation was also investigated using the epithelial cell model: MDCK cells. Depletion of anillin and SEPT9 in MDCK cells was achieved using lentivirus shRNA constructs and this revealed that anillin or SEPT9 depletion did not affect asymmetric cytokinesis, although localization of SEPT 9 was affected by anillin depletion.
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Staphylococcus aureus stimulates the release of constitutive tissue factor in lung epithelial cellsDeWalt, Robin I. 08 July 2011 (has links)
Sepsis is a life threatening condition caused by infectious agents, including the Gram-positive bacterium Staphylococcus aureus. Symptoms of sepsis often include intravascular coagulation and organ failure. Tissue factor (TF), the initiator of coagulation, may contribute to fibrin deposition in the lungs of patients with sepsis. We have found that lung epithelial cells constitutively express TF on the cell surface and in intracellular pools. Levels of TF diminished in response to S. aureus invasion possibly indicating a release in the form of shedding vesicles. TF levels diminish in response to viable bacteria, but not in response to heat killed (HK) bacteria. Our studies indicate that bacterial attachment at the host cell surface is insufficient to diminish levels of constitutive TF. Finally, we established that levels of constitutive intracellular TF diminish in response to the bacterial toxin, α-hemolysin, alone. This approach may provide a basis for understanding the role of TF in coagulation seen in sepsis. / Department of Biology
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Time heals all wounds? : mathematical models of epithelial and dermal wound healingDale, Paul David January 1995 (has links)
The mechanisms responsible for the healing of corneal surface wounds are the subject of biological controversy. In particular, the role and source of the regulatory chemical epidermal growth factor (EGF) is an area of intense debate. In the first part of this thesis, we propose a reaction-diffusion model which focuses on the stimulus for increased mitotic and migratory activity due to secretion of EGF. A detailed numerical study of various possible models, with parameter values based on biological data, reveals that, for realistic healing times, EGF must be released by the underlying layers of the cornea, in addition to the tear film source. The model exhibits travelling wave solutions and further analysis elucidates the interaction and role of the parameters in determining the speed of healing. Furthermore, we consider the effect of topical application of EGF and investigate the effect of curvature of the eye. We show that our model is consistent with many of the key features of corneal wound healing. Adult dermal wounds, in contrast to foetal wounds, heal with the formation of scar tissue. A crucial factor in determining the nature of the healed tissue is the ratio of collagen 1 to collagen 3, which indicates the fibril diameter. We develop a reaction-diffusion model which focuses on the stimulus for collagen synthesis due to the secretion of the different isoforms of the regulatory chemical transforming growth factor β (TGFβ). Numerical simulations of the model without diffusion lead to a value of this ratio consistent with that of healthy tissue for the foetus but corresponding to scarring in the adult. The model equations evolve to waves moving into the wound, but addition of TGFβ only has a transient effect on the final collagen levels. We investigate this effect by developing a caricature model. The model indicates that the main source of the fibroblasts is the underlying subcutaneous tissue and we determine key parameters which explain the difference between adult and foetal wound healing. Furthermore we make clinically testable predictions on the effects that topical application of various chemicals will have on scar formation.
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