Thesis (Ph.D.)--Indiana University, 2009. / Title from screen (viewed on December 1, 2009). Department of Cellular and Integrative Physiology, Indiana University-Purdue University Indianapolis (IUPUI). Advisor(s): B. Paul Herring, Anthony B. Firulli, Frederick M. Pavalko, Simon J. Rhodes. Includes vitae. Includes bibliographical references (leaves 138-149).

Glucan phosphate is a water‐soluble, yeast‐derived, (1 → 3)‐β‐D‐glucopyranose polymer that has been demonstrated to be a potent immune stimulant. Glucan phosphate administration is associated with stimulation of immunity and a concomitant increase in resistance to a variety of experimentally induced disease states. Preclinical safety evaluation indicates that glucan phosphate does not induce mortality or significant toxicity over a wide dose range. Phase I clinical data indicate that glucan phosphate will ameliorate immunosuppression, stimulate immunity, decrease susceptibility to infection and alter ultimate outcome in trauma patients following laparotomy or thoracotomy. This work reviews the development, characterization, preclinal and clinical evaluation of glucan phosphate.

No description available.

La transfusion sanguine est une thérapeutique indispensable pour laquelle il n’existe pas actuellement de substitut. La transfusion de produits sanguins labiles est dans la grande majorité des cas très bien tolérée mais elle peut être à l’origine d’effets indésirables chez les receveurs (EIR) notamment de type inflammatoire. Ceci dépend de facteurs liés aux produits eux-mêmes et/ou aux receveurs de par leur prédisposition génétique et de leur état clinique. Les concentrés plaquettaires (CP) sont la principale source de manifestations inflammatoires et/ou allergiques. Ceci est notamment dû, en partie, à la capacité des plaquettes à sécréter une multitude de molécules ayant une activité inflammatoire. De plus, les processus de collecte, de préparation et de conservation induisent un stress vis-à-vis des cellules, qui peut activer les plaquettes et donc induire la production de produits inflammatoires dans les CP. Le but de ce travail de thèse a été dans un premier temps d’identifier les molécules les plus impliquées dans les manifestations inflammatoires. Le sCD40L en particulier est identifié comme étant largement impliqué dans les EIR après transfusion de CP, mais pas systématiquement. Aussi, la composante inflammatoire de ces réactions est multifactorielle. De plus, nous avons évalué le potentiel inflammatoire des CP sur l’endothélium vasculaire. Des différences d’activation des cellules endothéliales, dans un modèle in vitro, ont été observées lorsqu’elles sont en présence de surnageants de CP ayant induits un EIR. Ce travail de thèse poursuit l’effort entrepris par notre équipe de recherche, en vue de prédire la survenue d’EIR et de préciser les mécanismes qui influencent la physiopathologie plaquettaire transfusionnelle ; un corollaire de ces travaux est ainsi d’optimiser les processus de production et de conditionnement des CP transfusés afin de réduire ces réactions inflammatoires. / Blood transfusion is an indispensable therapy for which there is currently no substitute. Transfusion of blood products is in the great majority of cases very well tolerated but it can be at the origin of serious adverse reactions (SARs), notably of inflammatory reactions. This depends on the factors related to the products themselves and/or to the recipients, their genetic predisposition and clinical condition. Platelet concentrates (PCs) are the main source of inflammatory and/or allergic manifestations. This is due, in part, to the ability of platelets to secrete a multitude of molecules with inflammatory activity. In addition, the collection, processing and storage conditions induce stress on cells, which can activate platelets and thus induce the production of inflammatory products in PCs. The purpose of this work is to identify the molecules involved in inflammatory manifestations. sCD40L was identified as being involved in SARs after PCs transfusion, but not systematically. Also, the inflammatory component of these reactions is multifactorial. In addition, we evaluated the inflammatory potential of PCs on the vascular endothelium. Differences in endothelial cell activation, in an in vitro model, were observed when they were in the presence of PC supernatants involved in SARs. This thesis work continues the effort undertaken by our research team to predict the occurrence of SARs and to clarify the mechanisms that influence transfusional platelet physiopathology; a corollary of this work is to optimize the production and conditioning process of PCs transfused in order to reduce these inflammatory reactions.

Master of Medicine / Non-melanoma skin cancer (NMSC) is the most common human cancer worldwide. Squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) make up almost all NMSC. SCC usually arises from actinic keratosis (AK) as a result of exposure to sunlight. SCC and AK provide a useful clinical model to investigate changes involved in the progression of NMSC. This project examines the expression of Brm, Brg-1, Snail 1 and Snail 2 in the progression of NMSC. Brm and Brg-1 are subunits of the SWI/SNF chromatin-remodelling complex which is involved in regulating the access of cell machinery to DNA by altering the structure of chromatin. It has been suggested that loss of this function is involved in carcinogenesis as the cell is unable to access to DNA normally in order to repair mutations or activate apoptosis. The loss of Brm or Brg-1 has been described in several human cancers. Snail 1 and Snail 2 are zinc-finger transcription factors that are known for their role in epithelial to mesenchymal transition (EMT), a process vital to embryological development. Increased expression of these factors leads to a loss of cell-cell adhesion and a migratory phenotype and has been described in some human cancers. In this project, double-label immunohistochemistry was used to determine the relative expression of these proteins in human SCC, BCC, AK and normal skin. The expression of Snail was unable to be determined due to poor specificity of the antibodies used. The expression of both Brm and Brg-1 proteins was found to be dramatically and consistently decreased in SCC and BCC when compared to normal skin and AK. This loss of Brm and Brg-1 occured as the tumour progressed from benign AK to malignant SCC. This finding suggests that the loss of either Brm or Brg-1 constitutes a key step in carcinogenesis. The results of this study identify Brm and Brg-1 as putative tumour suppressors involved in the progression of non-melanoma skin cancer from benign to malignant.

No description available.

Master of Medicine / Non-melanoma skin cancer (NMSC) is the most common human cancer worldwide. Squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) make up almost all NMSC. SCC usually arises from actinic keratosis (AK) as a result of exposure to sunlight. SCC and AK provide a useful clinical model to investigate changes involved in the progression of NMSC. This project examines the expression of Brm, Brg-1, Snail 1 and Snail 2 in the progression of NMSC. Brm and Brg-1 are subunits of the SWI/SNF chromatin-remodelling complex which is involved in regulating the access of cell machinery to DNA by altering the structure of chromatin. It has been suggested that loss of this function is involved in carcinogenesis as the cell is unable to access to DNA normally in order to repair mutations or activate apoptosis. The loss of Brm or Brg-1 has been described in several human cancers. Snail 1 and Snail 2 are zinc-finger transcription factors that are known for their role in epithelial to mesenchymal transition (EMT), a process vital to embryological development. Increased expression of these factors leads to a loss of cell-cell adhesion and a migratory phenotype and has been described in some human cancers. In this project, double-label immunohistochemistry was used to determine the relative expression of these proteins in human SCC, BCC, AK and normal skin. The expression of Snail was unable to be determined due to poor specificity of the antibodies used. The expression of both Brm and Brg-1 proteins was found to be dramatically and consistently decreased in SCC and BCC when compared to normal skin and AK. This loss of Brm and Brg-1 occured as the tumour progressed from benign AK to malignant SCC. This finding suggests that the loss of either Brm or Brg-1 constitutes a key step in carcinogenesis. The results of this study identify Brm and Brg-1 as putative tumour suppressors involved in the progression of non-melanoma skin cancer from benign to malignant.

There is an increasing evidence that alterations in chromatin remodeling play an important role in tumorigenesis. The SWI/SNF chromatin remodeling complexes contribute to the regulation of gene expression by altering the local chromatin structure. Depending on the context, they can act as either transcriptional activators or repressors. All SWI/SNF subcomplexes contain one of two ATPase subunits, Brm (Brahma) or Brg1 (Brahma related gene 1), which provide the energy for remodeling. Malignant melanoma is an aggressive cancer and is known for its notorious resistance to conventional anticancer therapies. MITF (microphthalmia-associated transcription factor) plays an essential role in melanoma biology and is placed on the central crossroad in the regulation of melanocyte development, differentation, maintenance of lineage identity, and survival of both normal and malignant melanocytes. Our results show that the active SWI/SNF complex is strictly required for the expression of MITF. This complex is also required for expression of some transcriptional MITF targets. The survival of melanoma cells is absolutely dependent on functional SWI/SNF complex and all subunits of this complex are expressed at high levels in melanoma cell lines. Primarily, Brg1-containing subcomplexes are more important for MITF...

Human error has been a key factor in grounding- and collision incidents in the past and continues to present a challenge to the maritime industry in the present day. The feasibility of a pilot framework for Bridge Resource Management (BRM) training lies at the heart of this study, its objective being to gauge the possibility of the construction of such a model. The result is thought to open up for further research and highlight the most relevant and effective BRM tools that can be used, by the bridge team, to prevent collision and grounding. The study was carried out in two stages. Firstly, four key personnel within the shipping industry were interviewed. These respondents were tasked with prioritising what they regarded as the most effective BRM tools. Secondly, the effectiveness of these tools was tested on marine incident cases. A document analysis was carried out on six collision and grounding cases, as investigated by the Marine Accident Investigation Branch (MAIB). The majority of the respondents were in agreement regarding which three BRM tools they regarded as the most relevant and effective. The result from the interviews was repeated in the result from the document analysis.  The conclusions that were drawn are that: it is possible to construct a pilot framework for BRM training and it is also possible to measure the effectiveness of the BRM tools. Additionally, suggestions for further research that arose from this thesis pertain to the study of the relationship between different BRM tools.

Indiana University-Purdue University Indianapolis (IUPUI) / There are many clinical diseases involving abnormal differentiation of smooth muscle, such as atherosclerosis, hypertension and asthma. In these diseases, one important pathological process is the disruption of the balance between differentiation and proliferation of smooth muscle cells. Serum Response Factor (SRF) has been shown to be a key regulator of smooth muscle differentiation, proliferation and migration through its interaction with various accessory proteins. Myocardin Related Transcrition Factors (MRTFs) are important co-activators of SRF that induce smooth muscle differentiation. Elucidating the mechanism of how MRTFs and SRF discriminate between genes required to regulate smooth muscle differentiation and those regulating proliferation will be a significant step toward finding a cure for these diseases. We hypothesized that SWI/SNF ATPdependent chromatin remodeling complexes, containing Brg1 and Brm, may play a role in this process. Results from western blotting and quantitative reverse transcription - polymerase chain reaction (qRT-PCR) analysis demonstrated that expression of dominant negative Brg1 or knockdown of Brg1 with silence ribonucleic acid (siRNA) attenuated expression of SRF/MRTF dependent smooth muscle-specific genes in primary cultures of smooth muscle cells. Immunoprecipitation assays revealed that Brg1, SRF and MRTFs form a complex in vivo and that Brg1 directly binds MRTFs, but not SRF, in vitro. Results from chromatin immunoprecipitation assays demonstrated that dominant negative Brg1 significantly attenuated SRF binding and the ability of MRTFs to increase SRF binding to the promoters of smooth muscle-specific genes, but not proliferation-related early response genes. The above data suggest that Brg1/Brm containing SWI/SNF complexes play a critical role in differentially regulating expression of SRF/MRTF-dependent genes through controlling the accessibility of SRF/MRTF to their target gene promoters. To examine the role of SWI/SNF in smooth muscle cells in vivo, we have generated mice harboring a smooth muscle-specific knockout of Brg1. Preliminary analysis of these mice revealed defects in gastrointestinal (GI) development, including a significantly shorter gut in Brg1 knockout mice. These data suggest that Brg1-containing SWI/SNF complexes play an important role in the development of the GI tract.