The effect of ionic redistributions on the microwave dielectric response of cytosol water upon glucose uptake.

The sensitivity of cytosol water's microwave dielectric (MD) response to D-glucose uptake in Red Blood Cells (RBCs) allows the detailed study of cellular mechanisms as a function of controlled exposures to glucose and other related analytes like electrolytes. However, the underlying mechanism behind the sensitivity to glucose exposure remains a topic of debate. In this research, we utilize MDS within the frequency range of 0.5-40 GHz to explore how ionic redistributions within the cell impact the microwave dielectric characteristics associated with D-glucose uptake in RBC suspensions. Specifically, we compare glucose uptake in RBCs exposed to the physiological concentration of Ca2+ vs. Ca-free conditions. We also investigate the potential involvement of Na+/K+ redistribution in glucose-mediated dielectric response by studying RBCs treated with a specific Na+/K+ pump inhibitor, ouabain. We present some insights into the MD response of cytosol water when exposed to Ca2+ in the absence of D-glucose. The findings from this study confirm that ion-induced alterations in bound/bulk water balance do not affect the MD response of cytosol water during glucose uptake.

Microwave Dielectric Relaxation of Univalent and Bivalent Electrolyte Solutions.

The microwave dielectric relaxation of aqueous solutions of univalent (KCl, NaCl, NaI) and bivalent (CaCl2, MgCl2) electrolytes at concentrations between 0.1 and 1 M at 25 °C was investigated using a vector network analyzer (0.5≤ ν ≤ 40 GHz). The spectra of these electrolyte systems are characterized by a symmetrical broadening of the main relaxation peak and were fitted using the Cole-Cole equation. In our analysis, we provide insights into the underlying physics of the relaxation events at microscopic and mesoscopic scales by using a 3D phase space trajectory that is based on the interactions of the relaxing dipole units with their surroundings and Frohlich's B function. The effect of the solutes on the H-bond network of water with increasing concentration is evident in the microwave dielectric spectra through decreasing dielectric strengths and relaxation times. It was found that the number of perturbed water molecules is higher in the case of bivalent electrolytes and appears to be proportional to the ionic radius. In our approach, the particular dependence between the broadening parameter α and the relaxation times τ reflects the rate of interactions between the elementary dipole units and their surroundings. We provide a quantitative analysis of the level of perturbation caused by the presence of ions in the hydrogen-bond network of water. It was found that the H-bonded network of water is highly perturbed in univalent systems compared to bivalent systems due to weaker bonded hydration shells. Finally, we found significant differences between the dielectric response of NaCl and NaI. The differences, originating in the counterions Cl- and I-, which are characterized by large ionic radii and consequently weaker electric fields in their vicinity, confirm that the effect of weakly hydrated ions should not be neglected in microwave dielectric spectra analysis.

The Impact of Ca2+ on Intracellular Distribution of Hemoglobin in Human Erythrocytes.

The membrane-bound hemoglobin (Hb) fraction impacts red blood cell (RBC) rheology and metabolism. Therefore, Hb-RBC membrane interactions are precisely controlled. For instance, the signaling function of membrane-bound deoxy-Hb and the structure of the docking sites in the cytosolic domain of the anion exchanger 1 (AE-1) protein are well documented; however, much less is known about the interaction of Hb variants with the erythrocyte's membrane. Here, we identified factors other than O2 availability that control Hb abundance in the membrane-bound fraction and the possible variant-specific binding selectivity of Hb to the membrane. We show that depletion of extracellular Ca2+ by chelators, or its omission from the extracellular medium, leads to membrane-bound Hb release into the cytosol. The removal of extracellular Ca2+ further triggers the redistribution of HbA0 and HbA2 variants between the membrane and the cytosol in favor of membrane-bound HbA2. Both effects are reversible and are no longer observed upon reintroduction of Ca2+ into the extracellular medium. Fluctuations of cytosolic Ca2+ also impact the pre-membrane Hb pool, resulting in the massive transfer of Hb to the cellular cytosol. We hypothesize that AE-1 is the specific membrane target and discuss the physiological outcomes and possible clinical implications of the Ca2+ regulation of the intracellular Hb distribution.

The inhibition of glucose uptake to erythrocytes: microwave dielectric response.

Dielectric spectroscopy has been used in the study and development of non-invasive glucose monitoring (NIGM) sensors, including the range of microwave frequencies. Dielectric relaxation of red blood cell (RBC) cytosolic water in the microwave frequency band has been shown to be sensitive to variations in the glucose concentration of RBC suspensions. It has been hypothesized that this sensitivity stems from the utilization of D-glucose by RBCs. To verify this proposition, RBCs were pretreated with inhibitors of D-glucose uptake (cytochalasin B and forskolin). Then their suspensions were exposed to different D-glucose concentrations as measured by microwave dielectric spectroscopy (MDS) in the 500 MHz-40 GHz frequency band. After incubation of RBCs with either inhibitor, the dielectric response of water in the cytoplasm, and specifically its relaxation time, demonstrated minimal sensitivity to the change of D-glucose concentration in the medium. This result allows us to conclude that the sensitivity of MDS to glucose uptake is associated with variations in the balance of bulk and bound RBC cytosolic water due to intracellular D-glucose metabolism, verifying the correctness of the initial hypothesis. These findings represent a further argument to establish the dielectric response of water as a marker of glucose variation in RBCs.