No description available.

No description available.

Most studies of insect Malpighian tubules (MTs) have examined transport of monovalent ions (K^+, Na^+, Cl^-). Isolated Drosophila melanogaster MTs also transport Ca^2+ from bath to lumen and transport is stimulated by cAMP. The lower segment of the MTs transports Ca^2+ at a higher rate per unit length than does the main segment known to produce the primary urine. This study examines both whole animal calcium regulation in larvae, pupae and adults and the mechanisms of Ca^2+ transport by isolated MTs. Drosophila melanogaster appears to regulate its calcium content and haemolymph calcium level. Calcium content of the whole fly only increased 10% with a 6.2-fold increase in dietary calcium. Anterior MTs can contain as much as 50% of the whole animal calcium content. The difference in MTs accumulation is due primarily to the enlarged initial segment of the anterior MTs. This segment, absent from the posterior MT, contains calcium-containing concretions. Whole fly calcium content does not increase continuously with the age implying that calcium is eventually being excreted. Haemolymph calcium concentrations do not change in response to changes in dietary calcium, suggesting that calcium concentration is regulated either by the rate of absorption or by the rate of excretion. The midgut and the enlarged initial segment of the anterior MTs may play important roles in haemolymph calcium regulation. Isolated MTs show sensitivity to both Ca^2+ channel blockers and Ca^2+ -ATPAse inhibitors on the basolateral and apical membranes respectively. Voltage-gated calcium channels appear to mediate calcium movement from bath to cell. A ruthenium red sensitive Ca^2+ -ATPAse may be used to transport calcium against the electrochemical gradient from cell to lumen. Lastly, the dissolution of luminal concretions plays a large role in net calcium secretion. / Thesis / Master of Science (MSc)

No description available.

The correct function of cardiac sarcomeric proteins allow for people to maintain quality of life. However, mutations of the cardiac sarcomeric proteins can result in remodeling of the heart which typically results in death. I present a full atomistic cardiac thin filament model that I have developed and three studies that I conducted while at the University of Arizona, while pursuing my doctoral degree in chemistry The goal was to develop the model to be able to study the effects of the mutations on the thin filament proteins. First, I present the long process of developing the model that is still evolving as new information is available. Second, I present the study of two mutants, the troponin T R92L mutant and the tropomyosin D230N mutant. Molecular dynamics was used to simulate the wild–type and mutant versions of the model which resulted in a visualization of the change of interaction between the tropomyosin and troponin, specifically at the overlap region. Third, I present the study of calcium release which is the "gatekeeper" to cardiac contraction. Steered molecular dynamics was utilized to find a previously unseen molecular mechanism that alters the rate of calcium release depending on the mutant. Fourth, I present the study of the mechanism of the tropomyosin transition across the actin filament, in which a longitudinal transition is favored. The studies helped to provide an atomistic level understanding of the cardiac thin filament as well as the methodology to which the mutations disrupt the natural functions of the sarcomeric proteins. The new results of the research can provide new insight into how the effects of the disease causing mutations can be mitigated, potentially extending the life of people with the conditions.

Calcium (Ca) homeostasis is the maintenance of a stable plasma Ca concentration in the human body in the presence of Ca variability in the physiological environment (e.g. by ingestion and/or excretion). For normal physiological function, the total plasma Ca concentration must be maintained within a very narrow range (2.2-2.4mM). Meeting such stringent requirements is the task of a regulatory system that employs parathyroid hormone (PTH) and calcitriol (CTL) to regulate Ca flux between the plasma and the kidneys, intestines and bones. On the other hand, plasma phosphate control is less tightly, but simultaneously, regulated via the same hormonal actions. Chronic imbalances in plasma Ca levels are associated with disorders of the regulatory organs, which cause abnormal hormonal secretion and activity. These changes in hormonal activity may lead to long-term problems, such as, osteoporosis (increased loss of bone mineral density), which arises from primary hyperparathyroidism (PHPT) – hyper secretion of PTH. Existing in silico models of Ca homeostasis in humans are often cast in the form of a single monolithic system of differential equations and are not easily amenable to the sort of tractable quantitative analysis from which one can acquire useful fundamental insight. In this research, the regulatory systems of plasma Ca and plasma phosphate are represented as an engineering control system where the physiological sub-processes are mapped onto corresponding block components (sensor, controller, actuator and process) and underlying mechanisms are represented by differential equations. Following validation of the overall model, Ca-related pathologies are successfully simulated through induced defects in the control system components. A systematic approach is used to differentiate PHPT from other diseases with similar pathophysiologies based on the unique hormone/ion responses to short-term Ca disturbance in each pathology model. Additionally, based on the changes in intrinsic parameters associated with PTG behavior, the extent of PHPT progression can be predicted and the enlarged gland size estimated a priori. Finally, process systems engineering methods are used to explore therapeutic intervention in two Ca-related pathologies: Primary (PHPT) and Secondary (SHPT) Hyperparathyroidism. Through parametric sensitivity analysis and parameter space exploration, the calcium-sensing receptor (sensor) is identified as a target site in both diseases and the extent of potential improvement is determined across the spectrum of severity of PHPT. The findings are validated against existing drug therapy, leading to a method of predicting drug dosage for a given stage of PHPT. Model Predictive Control is used in drug therapy in SHPT to customize the drug dosage for individual patients given the desired PTH outcome, and drug administration constraints. / Ph. D.