An experimental and theoretical analysis of nitric oxide availability in the microcirculation

Nitric Oxide (NO) is produced in the vascular endothelium where it then diffuses to the adjacent smooth muscle cells (SMC) activating agents known to regulate vascular tone. The close proximity of the site of NO production to the red blood cells (RBC) and its known fast consumption by hemoglobin, suggests that the blood will scavenge most of the NO produced. Therefore, it is unclear how NO is able to play its role in accomplishing vasodilation. Investigation of NO production and consumption rates will allow insight into this paradox. DAF-FM is a sensitive NO fluorescence probe widely used for qualitative assessment of cellular NO production. With the aid of a mathematical model of NO/DAF-FM reaction kinetics, experimental studies were conducted to calibrate the fluorescence signal showing that the slope of fluorescent intensity is proportional to [NO]2 and exhibits a saturation dependence on [DAF-FM]. In addition, experimental data exhibited a Km dependence on [NO]. This finding was incorporated into the model elucidating NO 2 as the possible activating agent of DAF-FM. A calibration procedure was formed and applied to agonist stimulated cells, providing an estimated NO release rate of 0.418 ± 0.18 pmol/cm2s. To assess NO consumption by RBCs, measurements of the rate of NO consumption in a gas stream flowing on top of an RBC solution of specified Hematocrit (Hct) was performed. The consumption rate constant (kbl)in porcine RBCs at 25°C and 45% Hct was estimated to be 3500 + 700 s-1. kbl is highly dependent on Hct and can reach up to 9900 + 4000 s-1 for 60% Hct. The nonlinear dependence of kbl on Hct suggests a predominant role for extracellular diffusion in limiting NO uptake. Further simulations showed a linear relationship between varying NO production rates and NO availability in the SMCs utilizing the estimated NO consumption rate. The corresponding SMC [NO] level for the average NO production rate estimated was approximately 15.1 nM. With the aid of experimental and theoretical methods we were able to examine the NO paradox and exhibit that endothelial derived NO is able to escape scavenging by RBCs to diffuse to the SMCs.

Dynamic Behavior and Fatigue Life of Highway Bridges Due to Doubling Heavy Vehicles

An increase in the demand for the freight shipping in the United States has been predicted for the near future and Longer Combination Vehicles (LCVs), which can carry more loads in each trip, seem like a good solution for the problem. Currently, utilizing LCVs is not permitted in most states of the US and little research has been conducted on the effects of these heavy vehicles on the roads and bridges. In this research, efforts are made to study these effects by comparing the dynamic and fatigue effects of LCVs with more common trucks. Ten Steel and prestressed concrete bridges with span lengths ranging from 30’ to 140’ are designed and modeled using the grid system in MATLAB. Additionally, three more real bridges including two single span simply supported steel bridges and a three span continuous steel bridge are modeled using the same MATLAB code. The equations of motion of three LCVs as well as eight other trucks are derived and these vehicles are subjected to different road surface conditions and bumps on the roads and the designed and real bridges. By forming the bridge equations of motion using the mass, stiffness and damping matrices and considering the interaction between the truck and the bridge, the differential equations are solved using the ODE solver in MATLAB and the results of the forces in tires as well as the deflections and moments in the bridge members are obtained. The results of this study show that for most of the bridges, LCVs result in the smallest values of Dynamic Amplification Factor (DAF) whereas the Single Unit Trucks cause the highest values of DAF when traveling on the bridges. Also in most cases, the values of DAF are observed to be smaller than the 33% threshold suggested by the design code. Additionally, fatigue analysis of the bridges in this study confirms that by replacing the current truck traffic with higher capacity LCVs, in most cases, the remaining fatigue life of the bridge is only slightly decreased which means that taking advantage of these larger vehicles can be a viable option for decision makers.