Search

Neuronal cell dysfunction plays an important role in neurodegenerative diseases. Oxidative stress can disrupt the redox balance within neuronal cells and may cause neuronal nitric oxide synthase (nNOS) to uncouple, contributing to the neurodegenerative processes. Experimental studies and clinical trials using nNOS cofactor tetrahydrobiopterin (BH 4 ) and antioxidants in neuronal cell dysfunction have shown inconsistent results. A better mechanistic understanding of complex interactions of nNOS activity and oxidative stress in neuronal cell dysfunction is needed. In this study, we developed a computational model of neuronal cell using nNOS biochemical pathways to explore several key mechanisms that are known to influence neuronal cell redox homeostasis. We studied the effects of oxidative stress and BH 4 synthesis on nNOS nitric oxide production and biopterin ratio (BH 4 /total biopterin). Results showed that nNOS remained coupled and maintained nitric oxide production for oxidative stress levels less than 230 nM/s. The results showed that neuronal oxidative stress above 230 nM/s increased the degree of nNOS uncoupling and introduced instability in the nitric oxide production. The nitric oxide production did not change irrespective of initial biopterin ratio of 0.05-0.99 for a given oxidative stress. Oxidative stress resulted in significant reduction in BH 4 levels even when nitric oxide production was not affected. Enhancing BH 4 synthesis or supplementation improved nNOS coupling, however the degree of improvement was determined by the levels of oxidative stress and BH 4 synthesis. The results of our mechanistic analysis indicate that there is a potential for significant improvement in neuronal dysfunction by simultaneously increasing BH 4 levels and reducing cellular oxidative stress., Competing Interests: Declaration of competing interest There is no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Red blood cells (RBCs) modulate nitric oxide (NO) bioavailability in the vasculature. Extracellular free hemoglobin (Hb) in the vascular lumen can cause NO bioavailability related complications seen in pathological conditions such as pancreatitis, sickle cell disease and malaria. In addition, the role of extracellular free Hb has been critical to estimate kinetic and transport properties of NO-RBCs interactions in 'competition experiments'. We recently reported a strong dependence of NO transport on RBC membrane permeability and hematocrit. NO donors combined with anti-inflammatory drugs are an emergent treatment for diseases like cancer, cardiovascular complications and wound healing. However, the role of RBCs in transport NO from NO donors is not clearly understood. To understand the significance of extracellular free Hb in pathophysiology on NO availability and estimation of the NO-RBC interactions, we developed a computational model to simulate NO biotransport to the RBC in the presence of extracellular free Hb. Using this model, we studied the effect of hematocrit, RBC membrane permeability and NO donors on NO-RBC interactions in the presence and absence of extracellular free Hb. The plasma NO concentration gradients and average plasma NO concentrations changed minimally with increase in extracellular free Hb concentrations at the higher hematocrit as compared to those at the lower hematocrit irrespective of the NO delivery method, indicating that the presence of extracellular free Hb affects the NO transport only at a low hematocrit. We also observed that NO concentrations increased with NO donor concentrations in the absence as well as in the presence of extracellular free Hb. In addition, NO donor supplementation may increase NO availability in the plasma in the event of loss of endothelium-derived NO activity., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Endothelial dysfunction causes an imbalance in endothelial NO and O₂·⁻ production rates and increased peroxynitrite formation. Peroxynitrite and its decomposition products cause multiple deleterious effects including tyrosine nitration of proteins, superoxide dismutase (SOD) inactivation, and tissue damage. Studies have shown that peroxynitrite formation during endothelial dysfunction is strongly dependent on the NO and O₂·⁻ production rates. Previous experimental and modeling studies examining the role of NO and O₂·⁻ production imbalance on peroxynitrite formation showed different results in biological and synthetic systems. However, there is a lack of quantitative information about the formation and biological relevance of peroxynitrite under oxidative, nitroxidative, and nitrosative stress conditions in the microcirculation. We developed a computational biotransport model to examine the role of endothelial NO and O₂·⁻ production on the complex biochemical NO and O₂·⁻ interactions in the microcirculation. We also modeled the effect of variability in SOD expression and activity during oxidative stress. The results showed that peroxynitrite concentration increased with increase in either O₂·⁻ to NO or NO to O₂·⁻ production rate ratio (QO₂·⁻/QNO or QNO/QO₂·⁻, respectively). The peroxynitrite concentrations were similar for both production rate ratios, indicating that peroxynitrite-related nitroxidative and nitrosative stresses may be similar in endothelial dysfunction or inducible NO synthase (iNOS)-induced NO production. The endothelial peroxynitrite concentration increased with increase in both QO₂·⁻/QNO and QNO/QO₂·⁻ ratios at SOD concentrations of 0.1-100 μM. The absence of SOD may not mitigate the extent of peroxynitrite-mediated toxicity, as we predicted an insignificant increase in peroxynitrite levels beyond QO₂·⁻/QNO and QNO/QO₂·⁻ ratios of 1. The results support the experimental observations of biological systems and show that peroxynitrite formation increases with increase in either NO or O₂·⁻ production, and excess NO production from iNOS or from NO donors during oxidative stress conditions does not reduce the extent of peroxynitrite mediated toxicity., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Nitric oxide (NO) consumption by red blood cell (RBC) hemoglobin (Hb) in vasculature is critical in regulating the vascular tone. The paradox of NO production at endothelium in close proximity of an effective NO scavenger Hb in RBCs is mitigated by lower NO consumption by RBCs compared to that of free Hb due to transport resistances including membrane resistance, extra- and intra-cellular resistances for NO biotransport to the RBC. Relative contribution of each transport resistance on NO-RBC interactions is still not clear. We developed a mathematical model of NO transport to a single RBC to quantify the contributions from individual transport barriers by analyzing the effect of RBC membrane permeability (P(m)), hematocrit (Hct) and NO-Hb reaction rate constants on NO-RBC interactions. Our results indicated that intracellular diffusion of NO was not a rate limiting step for NO-RBC interactions. The extracellular diffusion contributed 70-90% of total transport resistance for P(m)>1 cm s(-1) whereas membrane resistance accounts for 50-75% of total transport resistance for P(m)<0.1 cm s(-1). We propose a narrow P(m) range of 0.21-0.44 cm s(-1) for 10-45% Hct, respectively, below which membrane resistance is more significant and above which extracellular diffusion is a dominating transport resistance for NO-RBC interactions., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In sickle cell disease, the changes in RBC morphology destabilize the red blood cell (RBC) membrane and lead to hemolysis. Several experimental and clinical studies have associated intravascular hemolysis with pulmonary hypertension in sickle cell disease. Cell-free hemoglobin (Hb) from intravascular hemolysis has high affinity for nitrixc oxide (NO) and can affect the NO bioavailability in the sickle cell disease, which may eventually lead to pulmonary hypertension. To study the effects of intravascular hemolysis related cell-free Hb concentrations on NO bioavailability, we developed a two-dimensional mathematical model of NO biotransport in 50-μm arteriole under steady-state sickle cell disease conditions. We analyzed the effects of flow-dependent NO production and axial and radial transport of NO, a recently reported much lower NO-RBC reaction rate constant, and cell-free layer thickness on NO biotransport. Our results show that the presence of cell-free Hb concentrations as low as 0.5 μM results in an approximately three- to sevenfold reduction in the predicted smooth muscle cell NO concentrations compared with those under physiological conditions. In addition, increasing the diffusional resistance for NO in vascular lumen from cell-free layer or reducing NO-RBC reaction rate did not improve the NO bioavailability at the smooth muscle cell layer significantly for cell-free Hb concentrations ≥1 μM. These results suggest that lower NO bioavailability due to low micromolar cell-free Hb can disturb NO homeostasis and cause insufficient bioavailability at the smooth muscle cell layer. Our results supports the hypothesis that hemolysis-associated reduction in NO bioavailability may play a role in the development of pathophysiological complications like pulmonary hypertension in sickle cell disease that are observed in several clinical and experimental studies.

Endothelial dysfunction is associated with increase in oxidative stress and low NO bioavailability. The endothelial NO synthase (eNOS) uncoupling is considered an important factor in endothelial cell oxidative stress. Under increased oxidative stress, the eNOS cofactor tetrahydrobiopterin (BH(4)) is oxidized to dihydrobiopterin, which competes with BH(4) for binding to eNOS, resulting in eNOS uncoupling and reduction in NO production. The importance of the ratio of BH(4) to oxidized biopterins versus absolute levels of total biopterin in determining the extent of eNOS uncoupling remains to be determined. We have developed a computational model to simulate the kinetics of the biochemical pathways of eNOS for both NO and O(2)(•-) production to understand the roles of BH(4) availability and total biopterin (TBP) concentration in eNOS uncoupling. The downstream reactions of NO, O(2)(•-), ONOO(-), O(2), CO(2), and BH(4) were also modeled. The model predicted that a lower [BH(4)]/[TBP] ratio decreased NO production but increased O(2)(•-) production from eNOS. The NO and O(2)(•-) production rates were independent above 1.5μM [TBP]. The results indicate that eNOS uncoupling is a result of a decrease in [BH(4)]/[TBP] ratio, and a supplementation of BH(4) might be effective only when the [BH(4)]/[TBP] ratio increases. The results from this study will help us understand the mechanism of endothelial dysfunction., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The importance of nitric oxide (NO), superoxide (O2-), and peroxynitrite (ONOO-), interactions in physiologic functions and pathophysiological conditions such as cardiovascular disease, hypertension, and diabetes have been established extensively in in vivo and in vitro studies. Despite intense investigation of NO, O2-, and ONOO- biochemical interactions, fundamental questions regarding the role of these molecules remain unanswered. Mathematical models based on fundamental principles of mass balance and reaction kinetics have provided significant results in the case of NO. However, the models that include interaction of NO, O2-, and ONOO- have been few because of the complexity of these interactions. Not only do these mathematical and computational models provided quantitative knowledge of distributions and concentrations of NO, O2-, and ONOO- under normal physiologic and pathophysiologic conditions, they also can help to answer specific hypotheses. The focus of this review article is on the models that involve more than one of the 3 molecules (NO, O2-, and ONOO-). Specifically, kinetic models of O2- dismutase and tyrosine nitration and biotransport models in the microcirculation are reviewed. In addition, integrated experimental and computational models of dynamics of NO/O2-/ONOO- in diverse systems are reviewed.

Bioavailability of vasoactive endothelium-derived nitric oxide (NO) in vasculature is a critical factor in regulation of many physiological processes. Consumption of NO by RBC plays a crucial role in maintaining NO bioavailability. Recently, Deonikar and Kavdia (2009b) reported an effective NO-RBC reaction rate constant of 0.2×10(5)M(-1)s(-1) that is ~7 times lower than the commonly used NO-RBC reaction rate constant of 1.4×10(5)M(-1)s(-1). To study the effect of lower NO-RBC reaction rate constant and nitrite and nitrate formation (products of NO metabolism in blood), we developed a 2D mathematical model of NO biotransport in 50 and 200μm ID arterioles to calculate NO concentration in radial and axial directions in the vascular lumen and vascular wall of the arterioles. We also simulated the effect of blood velocity on NO distribution in the arterioles to determine whether NO can be transported to downstream locations in the arteriolar lumen. The results indicate that lowering the NO-RBC reaction rate constant increased the NO concentration in the vascular lumen as well as the vascular wall. Increasing the velocity also led to increase in NO concentration. We predict increased NO concentration gradient along the axial direction with an increase in the velocity. The predicted NO concentration was 281-1163nM in the smooth muscle cell layer for 50μm arteriole over the blood velocity range of 0.5-4cms(-1) for k(NO-RBC) of 0.2×10(5)M(-1)s(-1), which is much higher than the reported values from earlier mathematical modeling studies. The NO concentrations are similar to the experimentally measured vascular wall NO concentration range of 300-1000nM in several different vascular beds. The results are significant from the perspective that the downstream transport of NO is possible under the right circumstances., (Copyright © 2010 Elsevier Inc. All rights reserved.)

In blood vessels, nitric oxide homeostasis is maintained by its formation by endothelial nitric oxide synthase and its consumption in smooth muscle cells and in vascular lumen by red blood cell (RBC) encapsulated hemoglobin (Hb). Free hemoglobin has a very high reaction rate (k(Hb-NO) approximately 10(7) M(-1) s(-1)) with NO as compared to RBC-Hb. Mechanisms of reduced NO uptake by RBC-Hb has been extensively studied in recent years. A critical factor in the investigation of NO-RBC interactions is delivery of NO. Common NO delivery methods include use of NO donors and bolus saturated NO solutions, which delivers NO homogeneously and only in the vicinity of bolus, respectively. In this study, we developed a flow system that uses gaseous delivery of NO through a polymeric semi-permeable membrane to obtain precise and uniform NO concentrations for NO-RBC interactions. We conducted experiments using the flow system to study the effect of NO concentrations, hematocrit and RBC suspension flow rates on NO-RBC interactions. We developed a computational model to simulate NO transport and to estimate the reaction rate constant for NO-RBC interaction in the flow system. Our results showed that NO consumption of RBCs (i) increased linearly with an increase in available NO, and (ii) decreased with increase in RBCs suspension flow rate. We estimated the reaction rate constant for NO-RBC interactions to be 0.2 x 10(5) M(-1) s(-1) which is approximately 1250-fold lower than NO consumption by free hemoglobin and approximately 2.5-20 fold slower than reported NO-RBC reaction rate.

Nitric oxide (NO) is a potent vasodilator and its homeostasis depends on interaction with RBCs. A key factor in understanding NO-RBC interactions in vascular lumen is a comprehensive analysis of product identification and quantification. In this context, administration of NO during in vitro NO-RBC interactions becomes a crucial variable. In this study, we designed a bioreactor that maintains a precise NO concentration in the headspace that diffuses to RBCs suspension to study the quantitative effect of NO concentration and hematocrit (Hct) on NO-RBC interactions. The products of NO-RBC reaction (nitrite and total nitrogen species (total NOx)) were measured by chemiluminescence assay. A mathematical model simulating NO biotransport to a single RBC was developed to (1) estimate NO-RBC reaction rate constant; (2) predict the NO concentrations in the bulk RBC suspension and at the RBC membrane for RBC membrane NO permeability (P(m)) values of 0.0415-40 cm/s. Measured nitrite and total NOx concentrations increased with increase in headspace NO concentration whereas nitrite concentrations decreased with hematocrit and total NOx concentrations increased with hematocrit. This indicates that the extracellular resistance is a controlling factor for RBC uptake of NO. Modeling results showed that the effective reaction rate constant (k(eff)) for NO-RBC interactions was 2.32 x 10(4)-1.08 x 10(6) M(-1) s(-1). Results also predict that the membrane permeability in the range of 0.0415-0.4 cm/s is required to maintain physiologically relevant levels of NO at the smooth muscle cell layer. The effective reaction rate constant increased with increase in P(m) and magnitude of increase was higher at 45% Hct. For all P(m) values, the k(hb)/k(eff) ratios were lower for 45% Hct as compared to 5% Hct indicating extracellular resistance is important for RBC NO uptake. Our experimental and mathematical analyses of NO-RBC interactions indicate that both unstirred layer and RBC membrane have a significant effect on NO transport to RBCs. In addition, the membrane permeability in the range of 0.0415-0.4 cm/s is required to maintain sufficient NO concentrations at the smooth muscle cell layer., (Copyright 2009 Elsevier Inc. All rights reserved.)

Pathogenesis of many of diabetes-related vascular complications is associated with endothelial cell (EC) dysfunction, which is reduced bioavailability of EC-released nitric oxide (NO). Interaction dynamics of NO, superoxide (O(2)(-)) and peroxynitrite (ONOO(-)) are dependent on both their productions and consumptions through various pathways. Quantitative knowledge of these interaction dynamics in high glucose-induced EC dysfunction remains poorly understood. We developed an integrated experimental and computational approach to gain a quantitative understanding of the interactions of NO, O(2)(-) and ONOO(-) in high glucose-exposed ECs. End-products, nitrite and nitrate, were measured using a chemiluminescence analyzer. A computational biochemical reaction network model was developed to predict the effect of high glucose on ECs NO, O(2)(-) and ONOO(-). ECs NO and O(2)(-) productions increased in high glucose as evidenced by increased total NOx concentration, primarily increasing nitrate concentration. The model predicted an increase in O(2)(-) and ONOO(-) concentrations and a decrease in NO concentration in high glucose conditions. Administration of superoxide dismutase (SOD) decreased O(2)(-) concentration and increased NO concentration, thus SOD improved high glucose-induced changes in these interactions. An important finding of this study was that the NO bioavailability decreased in high glucose conditions even though NO production of EC increased. The integrated approach provides a framework to predict NO, O(2)(-) and ONOO(-) concentrations and productions that are difficult to measure in one experiment and will be useful in further EC dysfunction studies.

Interactions of free radicals such as superoxide (O2-), nitric oxide (NO), and peroxynitrite (ONOO-) are important in pathophysiological conditions such as hypertension, atherosclerosis, diabetes and the resulting cardiovascular diseases. Excessive levels of superoxide during oxidative stress cause a reduction in NO bioavailability by forming peroxynitrite and resulting in endothelial dysfunction. Superoxide dismutase (SOD) competes with NO for superoxide, and reduces the formation of peroxynitrite. In this study, we developed a mathematical model for free radical transport within and around an arteriolar vessel based on the fundamental principles of mass balance, reaction kinetics, and vascular geometry. We used the model to study the effect of the three types of SOD, viz. CuZn-SOD, Mn-SOD and extra cellular-SOD, on the bioavailability of NO. Results indicate that SOD location and concentration in the arteriole significantly affect superoxide concentration. The model predicts that a reduction in SOD levels results in increased superoxide and peroxynitrite concentrations and decreased NO concentration in the vessel. The results also suggest a role of SOD in the amelioration of oxidative stress and NO bioavailability in microcirculation. This model will help in furthering our knowledge of endothelial dysfunction in pathological conditions and the impact of specific SODs on free radical interactions.

Venular endothelial cells can release nitric oxide (NO) in response to intraluminal flow both in isolated venules and in vivo. Experimental studies suggest that venular endothelium-released NO causes dilation of the adjacent paired arteriole. In the vascular wall, NO stimulates its target hemoprotein, soluble guanylate cyclase (sGC), which relaxes smooth muscle cells. In this study, a computational model of NO transport for an arteriole and venule pair was developed to determine the importance of the venular endothelium-released NO and its transport to the adjacent arteriole in the tissue. The model predicts that the tissue NO levels are affected within a wide range of parameters, including NO-red blood cell reaction rate and NO production rate in the arteriole and venule. The results predict that changes in the venular NO production affected not only venular endothelial and smooth muscle NO concentration but also endothelial and smooth muscle NO concentration in the adjacent arteriole. This suggests that the anatomy of microvascular tissue can permit the transport of NO from arteriolar to venular side, and vice versa, and may provide a mechanism for dilation of proximal arterioles by venules. These results will have significant implications for our understanding of tissue NO levels in both physiological and pathophysiological conditions.

Nitric oxide (NO) plays an important role in autocrine and paracrine manner in numerous physiological processes, including regulation of blood pressure and blood flow, platelet aggregation, and leukocyte adhesion. In vascular wall, most of the bioavailable NO is believed to derive from endothelial cell NO synthase (eNOS). Recently, neuronal NOS (nNOS) has been identified as a source of NO in the vicinity of microvessels and has been shown to participate in vascular function. Thus NO can be produced and transported to the vascular smooth muscle cells from 1). endothelial cells and 2). perivascular nerve fibers, mast cells, and other nNOS-containing sources. In this study, a mathematical model of NO diffusion-reaction in a cylindrical arteriolar segment was formulated. The model quantifies the relative contribution of these NO sources and the smooth muscle availability of NO in a tissue containing an arteriolar blood vessel. The results indicate that a source of NO derived through nNOS in the perivascular region can be a significant contributor to smooth muscle NO. Predicted smooth muscle NO concentrations are as high as 430 nM, which is consistent with reported experimental measurements ( approximately 400 nM). In addition, we used the model to analyze the smooth muscle NO availability in 1). eNOS and nNOS knockout experiments, 2). the presence of myoglobin, and 3). the presence of cell-free Hb, e.g., Hb-based oxygen carriers. The results show that NO release by nNOS would significantly affect available smooth muscle NO. Further experimental and theoretical studies are required to account for distribution of NOS isoforms and determine NO availability in vasculatures of different tissues.

Nitric oxide (NO) plays many important physiological roles, including the regulation of vascular smooth muscle tone. In response to hemodynamic or agonist stimuli, endothelial cells produce NO, which can diffuse to smooth muscle where it activates soluble guanylate cyclase (sGC), leading to cGMP formation and smooth muscle relaxation. The close proximity of red blood cells suggests, however, that a significant amount of NO released will be scavenged by blood, and thus the issue of bioavailability of endothelium-derived NO to smooth muscle has been investigated experimentally and theoretically. We formulated a mathematical model for NO transport in an arteriole to test the hypothesis that transient, burst-like NO production can facilitate efficient NO delivery to smooth muscle and reduce NO scavenging by blood. The model simulations predict that 1) the endothelium can maintain a physiologically significant amount of NO in smooth muscle despite the presence of NO scavengers such as hemoglobin and myoglobin; 2) under certain conditions, transient NO release presents a more efficient way for activating sGC and it can increase cGMP formation severalfold; and 3) frequency-rather than amplitude-dependent control of cGMP formation is possible. This suggests that it is the frequency of NO bursts and perhaps the frequency of Ca(2+) oscillations in endothelial cells that may limit cGMP formation and regulate vascular tone. The proposed hypothesis suggests a new functional role for Ca(2+) oscillations in endothelial cells. Further experimentation is needed to test whether and under what conditions in silico predictions occur in vivo.

The endothelium-derived nitric oxide (NO) is one of the mediators of smooth muscle (SM) relaxation. The release of NO by endothelium depends on the wall shear stress (WSS) to which endothelium is exposed. During hemodilution or isovolemic exchange transfusion with hemoglobin-based oxygen carriers (HBOCs) or volume expanders, the systemic hematocrit, blood viscosity, and blood flow rate are affected that would change WSS at endothelium. The effect of WSS-dependent NO release on SM NO availability has not been determined by direct measurements. We have formulated a mathematical model that is capable of predicting NO concentration in and around arteriolar vessels. The model predicts that the normal physiological SM NO concentration is approximately 100 nM at a physiological WSS of 24 dyn/cm(2) and the NO concentration is linearly dependent on WSS. With volume expanders, the SM NO concentration increases significantly and the levels of SM NO are significantly higher with increase in WSS. The SM NO decreases several-fold even for 5 microM luminal HBOC. For HBOCs, the NO levels are not restored to normal physiological level even with a significant increase in WSS (>48 dyn/cm(2)). These predictions are consistent with the results of animal studies of vascular tone following administration of HBOCs and volume expanders.

As a small biological molecule, nitric oxide (NO), plays a key role in diverse functions including smooth muscle cell regulation, neurotransmission, inhibition of platelet aggregation, and cytotoxic actions. The assessment of NO effects in biological systems has extensively been studied using NO donor compounds that often have differing NO release mechanisms and kinetic rates. Due to the differing kinetic rates and release mechanisms, in addition to reactions involving NO (such as autoxidation of NO), the NO concentrations to which biological systems are exposed may vary significantly depending upon the NO donor compound. Thus, quantifying the effects of NO using different NO donors is difficult unless the NO concentration profile in the experimental system is predicted or measured. In this study, the spatial and temporal NO concentration in a stagnant system (such as a culture plate or micro-well) is modeled following the addition of an NO donor characterized with first-order NO release kinetics. Two NO donors were utilized: diethylamine NONOate (DEA/NO) and spermine NONOate (SPER/NO). The use of a mathematical model can eliminate the need of complex in situ NO measurements and be useful for predicting the physical loss of NO from the experimental system. In addition, properly scaling the NO concentration can be useful in estimating the maximum NO concentration that will exist in solution. The results show that under widely used in vitro experimental conditions, including varying NO donor concentrations, cellular oxygen consumption rates, and aqueous phase heights, the spatial and temporal NO concentration range can vary significantly. In addition, hypoxic conditions can occur in the vicinity of cells, and in some situations, the physical loss of NO from the experimental system may be significant.

Administration of hemoglobin-based oxygen carriers (HBOCs) frequently results in vasoconstriction that is primarily attributed to the scavenging of endothelium-derived nitric oxide (NO) by cell-free hemoglobin. The ensuing pressor response could be caused by the high NO reactivity of HBOC in the vascular lumen and/or the extravasation of hemoglobin molecules. There is a need for quantitative understanding of the NO interaction with HBOC in the blood vessels. We developed a detailed mathematical model of NO diffusion and reaction in the presence of an HBOC for an arteriolar-size vessel. The HBOC reactivity with NO and degree of extravasation was studied in the range of 2-58 x 10(6) M(-1) x s(-1) and 0-100%, respectively. The model predictions showed that the addition of HBOC reduced the smooth muscle (SM) NO concentration in the activation range (12-28 nM) for soluble guanylate cyclase, a major determinant of SM contraction. The SM NO concentration was significantly reduced when the extravasation of HBOC molecules was considered. The myoglobin present in the parenchymal cells scavenges NO, which reduces the SM NO concentration.

A plasma stratum (cell free layer or CFL) generated by flowing blood interposed between the red blood cell (RBC) core and the endothelium affects generation, consumption, and transport of nitric oxide (NO) in the microcirculation. CFL width is a principal factor modulating NO diffusion and vessel wall shears stress development, thus significantly affecting NO bioavailability. Since the CFL is bounded by the surface formed by the chaotically moving RBCs and the stationary but spatially non-uniform endothelial surface, its width fluctuates randomly in time and space. We analyze how these stochastic fluctuations affect NO transport in the CFL and NO bioavailability. We show that effects due to random boundaries do not average to zero and lead to an increase of NO bioavailability. Since endothelial production of NO is significantly enhanced by temporal variability of wall shear stress, we posit that stochastic shear stress stimulation of the endothelium yields the baseline continual production of NO by the endothelium. The proposed stochastic formulation captures the natural continuous and microscopic variability, whose amplitude is measurable and is of the scale of cellular dimensions. It provides a realistic model of NO generation and regulation. [ABSTRACT FROM AUTHOR]

Nitric oxide (NO) plays an important role in autocrine and paracrine manner in numerous physiological processes, including regulation of blood pressure and blood flow, platelet aggregation, and leukocyte adhesion. In vascular wall, most of the bioavailable NO is believed to derive from endothelial cell NO synthase (eNOS). Recently, neuronal NOS (nNOS) has been identified as a source of NO in the vicinity of microvessels and has been shown to participate in vascular function. Thus NO can be produced and transported to the vascular smooth muscle cells from 1) endothelial cells and 2) perivascular nerve fibers, mast cells, and other nNOS-containing sources. In this study, a mathematical model of NO diffusion-reaction in a cylindrical arteriolar segment was formulated. The model quantifies the relative contribution of these NO sources and the smooth muscle availability of NO in a tissue containing an arteriolar blood vessel. The results indicate that a source of NO derived through nNOS in the perivascular region can be a significant contributor to smooth muscle NO. Predicted smooth muscle NO concentrations are as high as 430 nM, which is consistent with reported experimental measurements (∼400 nM). In addition, we used the model to analyze the smooth muscle NO availability in 1) eNOS and nNOS knockout experiments, 2) the presence of myoglobin, and 3) the presence of cell-free Hb, e.g., Hb-based oxygen carriers. The results show that NO release by nNOS would significantly affect available smooth muscle NO. Further experimental and theoretical studies are required to account for distribution of NOS isoforms and determine NO availability in vasculatures of different tissues. [ABSTRACT FROM AUTHOR]

The endothelium-derived nitric oxide (NO) is one of the mediators of smooth muscle (SM) relaxation. The release of NO by endothelium depends on the wall shear stress (WSS) to which endothelium is exposed. During hemodilution or isovolemic exchange transfusion with hemoglobin-based oxygen carriers (HBOCs) or volume expanders, the systemic hematocrit, blood viscosity, and blood flow rate are affected that would change WSS at endothelium. The effect of WSS-dependent NO release on SM NO availability has not been determined by direct measurements. We have formulated a mathematical model that is capable of predicting NO concentration in and around arteriolar vessels. The model predicts that the normal physiological SM NO concentration is ∼100 nM at a physiological WSS of 24 dyn/cm2 and the NO concentration is linearly dependent on WSS. With volume expanders, the SM NO concentration increases significantly and the levels of SM NO are significantly higher with increase in WSS. The SM NO decreases several-fold even for 5 μM luminal HBOC. For HBOCs, the NO levels are not restored to normal physiological level even with a significant increase in WSS (>48 dyn/cm2). These predictions are consistent with the results of animal studies of vascular tone following administration of HBOCs and volume expanders. [Copyright &y& Elsevier]

All cells of vascular tissue including endothelial cells, and smooth muscle cells have enzyme systems that produce reactive oxygen species (ROS). Important ROS generated in the vascular tissue are superoxide or hydrogen peroxide. Superoxide can react with endothelium-derived nitric oxide (NO) to form peroxynitrite. Antioxidant systems protect against damage from ROS in normal physiological conditions. When ROS generation overwhelms the antioxidant defense (known as oxidative stress), these radicals can alter cellular function by interacting with DNA, RNA, and fatty acids, contribute to impairment of endothelium-dependent vasodilation and lead to apoptosis. We formulated a detailed computational model of NO, superoxide and peroxynitrite transport in a tissue containing an arteriolar blood vessel to quantify these species biochemical interactions and transport in the microcirculation. In this study, we analyzed compartmentalization and localization of superoxide formation and resulting tyrosine nitration formation. The model predicted that NO, superoxide and peroxynitrite levels and resulting tyrosine nitration are affected by a number of factors including the location and extent of oxidative stress. The model predictions provide insightinto role of individual forms of superoxide dismutase in physiologic and disease conditions. The results are significant because tyrosine nitration is observed in many pathological conditions but cellular sources for superoxide varies in different disease states. [ABSTRACT FROM AUTHOR]

Nitric oxide (NO) bioavailability is implicated in determining fate of many physiological and pathophysiological processes. In vasculature, nitric oxide homeostasis is maintained by its production by endothelial nitric oxide synthase and consumption by red blood cells (RBC) and leads to smooth muscle cells vasorelaxation. The reaction between NO and RBCs is under intense investigation. We studied the quantitative effects of oxygenation and hematocrit on NO -- RBC interactions. For this purpose, we designed a novel stirred-bioreactor with a headspace to maintain controlled NO gas concentration. Gaseous NO was reacted with oxygenated and deoxygenated RBC suspension for a period of 10 min and samples were taken every 2 min. To study the effect of hematocrit on NO -- RBC interactions, three hematocrit suspensions (5%, 20% and 45%) were reacted with NO for 10 min. Nitrite and total nitrogen species (total NOx) contents of all samples were measured by chemiluminescence assay and hemoglobin derivatives were measured using spectrophotometric analysis. Oxygenated NO-RBC interactions resulted in higher nitrite and total NOx concentrations than that of deoxygenated NO-RBC interactions. Nitrite concentrations decreased with hematocrit and total NOx concentrations increased with hematocrit. These results will enhance the present understanding of hematocrit and oxygenation effects on NO-RBC interactions. [ABSTRACT FROM AUTHOR]

Glutathione (GSH) and GSH/glutathione peroxidase (GPX) enzyme system is essential for normal intracellular homeostasis and gets disturbed under pathophysiologic conditions including endothelial dysfunction. Overproduction of reactive oxidative species (ROS) and reactive nitrogen species (RNS) including superoxide (O 2 •−), and the loss of nitric oxide (NO) bioavailability is a characteristic of endothelial dysfunction. The GSH/GPX system play an important role in eliminating ROS/RNS. Studies have provided important information regarding the interactions of ROS/RNS with the GSH/GPX in biological systems; however, it is not clear how this cross talk affect these reactive species and GSH/GPX enzyme system, under physiologic and oxidative/nitrosative stress conditions. In the present study, we developed a detailed endothelial cell kinetic model to understand the relationship amongst the key enzyme systems including GSH, GPX, peroxiredoxin (Prx) and reactive species, such as hydrogen peroxide (H 2 O 2), peroxynitrite (ONOO−), and dinitrogen trioxide (N 2 O 3). Our simulation results showed that the alterations in the generation rates of O 2 •− and NO led to the formation of a wide range of ROS and RNS. Simulations performed by varying the ratio of O 2 •− to NO generation rates as well as GSH and GPX concentrations showed that the GPX reducing capacity was dependent on GSH availability, level of oxidative/nitrosative stress, and can be attributed to N 2 O 3 levels, but not to H 2 O 2 and ONOO−. Our results showed that N 2 O 3 mediated switch-like depletion in GSH and the incorporation of Prx had no considerable effect on the ROS/RNS species other than ONOO− and H 2 O 2. The analysis presented in this study will improve our understanding of vascular diseases in which the levels and oxidation states of GSH, GPX and/or Prx are significantly altered and pharmacological interventions show limited benefits. • Computational model to analyze interactions of ROS/RNS with GSH/GPX is presented. • Recycling of GPX is attributed to N 2 O 3 availability but not to H 2 O 2 & ONOO−. • N 2 O 3 mediates GSH depletion in a switch-like manner. • Prx removes ONOO− while both Prx and GSH/GPX are needed for H 2 O 2 removal. • GSH/GPX activity is independent of physiologic NADPH levels. [ABSTRACT FROM AUTHOR]