Volatile Compound Markers in Beef Irradiated with Accelerated Electrons.

This study focuses on the behavior of volatile organic compounds in beef after irradiation with 1 MeV accelerated electrons with doses ranging from 0.25 kGy to 5 kGy to find reliable dose-dependent markers that could be used for establishing an effective dose range for beef irradiation. GC/MS analysis revealed that immediately after irradiation, the chemical yield and accumulation rate of lipid oxidation-derived aldehydes was higher than that of protein oxidation-derived aldehydes. The nonlinear dose-dependent relationship of the concentration of volatile organic compounds was explained using a mathematical model based on the simultaneous occurrence of two competing processes: decomposition of volatile compounds due to direct and indirect action of accelerated electrons, and accumulation of volatile compounds due to decomposition of other compounds and biomacromolecules. A four-day monitoring of the beef samples stored at 4 °C showed that lipid oxidation-derived aldehydes, protein oxidation-derived aldehydes and alkanes as well as alcohol ethanol as an indicator of bacterial activity were dose-dependent markers of biochemical processes occurring in the irradiated beef samples during storage: oxidative processes during direct and indirect action of irradiation, oxidation due to the action of reactive oxygen species, which are always present in the product during storage, and microbial-enzymatic processes. According to the mathematical model of the change in the concentrations of lipid oxidation-derived aldehydes over time in the beef samples irradiated with different doses, it was found that doses ranging from 0.25 kGy to 1 kGy proved to be most effective for beef irradiation with accelerated electrons, since this dose range decreases the bacterial content without considerable irreversible changes in chemical composition of chilled beef during storage.

Hemoglobin Derivatives in Beef Irradiated with Accelerated Electrons.

The efficiency of food irradiation depends on the accuracy of the irradiation dose range that is sufficient for inhibiting microbiological growth without causing an irreversible change to the physical and chemical properties of foods. This study suggests that the concentration of hemoglobin derivatives can be used as a criterion for establishing the limit for chilled beef irradiation at which irradiation-induced oxidation becomes irreversible. The express spectrophotometry method for estimating the hemoglobin derivative concentration shows a nonlinear increase in methemoglobin concentration from 15% to 50% in beef irradiated by accelerated electrons with the doses ranging from 250 Gy to 10,000 Gy. The monitoring of the hemoglobin derivative concentration for three days after irradiation shows nonmonotonous dependencies of methemoglobin concentration in beef in the storage time since the oxidation of hemoglobin occur as a result of irradiation and biochemical processes in beef during storage. The proposed method based on the quantitative analysis of the hemoglobin derivative concentration can be used to estimate the oxidation level for irradiation of foods containing red blood cells. The study proposes a model that describes the change in hemoglobin derivative concentration in beef after irradiation considering that oxidation of hemoglobin can be triggered by the direct ionization caused by accelerated electrons, biochemical processes as a result of bacterial activity, and reactive oxygen species appearing during irradiation and storage. This research throws light on the mechanisms behind food irradiation during storage that should be taken into account for selecting the optimal parameters of irradiation.

Atomic Force Microscopy and High-Resolution Spectrophotometry for Study of Anoxemia and Normoxemia in Model Experiment In Vitro.

The oxygen content in the blood may decrease under the influence of various physicochemical factors and different diseases. The state of hypoxemia is especially dangerous for critically ill patients. In this paper, we describe and analyze the changes in the characteristics of red blood cells (RBCs) with decreasing levels of oxygen in the RBC suspension from normoxemia to hypoxemia/anoxemia in an in vitro model experiment. The RBCs were stored in hypoxemia/anoxemia and normoxemia conditions in closed and open tubes correspondingly. For the quantitative study of RBC parameter changes, we used atomic force microscopy, digital spectrophotometry, and nonlinear curve fitting of the optical spectra. In both closed and open tubes, at the end of the storage period by day 29, only 2% of discocytes remained, and mainly irreversible types, such as microspherocytes and ghosts, were observed. RBC hemolysis occurred at a level of 25-30%. Addition of the storage solution, depending on the concentration, changed the influence of hypoxemia on RBCs. The reversibility of the change in hemoglobin derivatives was checked. Based on the experimental data and model approach, we assume that there is an optimal level of hypoxemia at which the imbalance between the oxidative and antioxidant systems, the rate of formation of reactive oxygen species, and, accordingly, the disturbances in RBCs, will be minimal.

Mechanochemical Synergism of Reactive Oxygen Species Influences on RBC Membrane.

The influences of various factors on blood lead to the formation of extra reactive oxygen species (ROS), resulting in the disruption of morphology and functions of red blood cells (RBCs). This study considers the mechanisms of the mechanochemical synergism of OH• free radicals, which are most active in the initiation of lipid peroxidation (LPO) in RBC membranes, and H2O2 molecules, the largest typical diffusion path. Using kinetic models of differential equations describing CH2O2t and COH•t, we discuss two levels of mechanochemical synergism that occur simultaneously: (1) synergism that ensures the delivery of highly active free radicals OH• to RBC membranes and (2) a positive feedback system between H2O2 and OH•, resulting in the partial restoration of spent molecules. As a result of these ROS synergisms, the efficiency of LPO in RBC membranes sharply increases. In blood, the appearance of OH• free radicals is due to the interaction of H2O2 molecules with free iron ions (Fe2+) which arise as a result of heme degradation. We experimentally established the quantitative dependences of COH• CH2O2 using the methods of spectrophotometry and nonlinear curve fitting. This study extends the analysis of the influence of ROS mechanisms in RBC suspensions.

Structural Configuration of Blood Cell Membranes Determines Their Nonlinear Deformation Properties.

The ability of neutrophils and red blood cells (RBCs) to undergo significant deformations is a key to their normal functioning. Disruptions of these processes can lead to pathologies. This work studied the influence of structural configuration rearrangements of membranes after exposure to external factors on the ability of native membranes of neutrophils and RBCs to undergo deep deformation. The rearrangement of the structural configuration of neutrophil and RBC membranes under the influence of cytological fixatives caused nonlinear deformation phenomena. There were an increase in Young's modulus, a decrease in the depth of homogeneous bending, and a change in the distance between cytoskeletal junctions. Based on the results of the analysis of experimental data, a mathematical model was proposed that describes the process of deep bending of RBСs and neutrophil membranes.

Investigation of Red Blood Cells by Atomic Force Microscopy.

Currently, much research is devoted to the study of biological objects using atomic force microscopy (AFM). This method's resolution is superior to the other non-scanning techniques. Our study aims to further emphasize some of the advantages of using AFM as a clinical screening tool. The study focused on red blood cells exposed to various physical and chemical factors, namely hemin, zinc ions, and long-term storage. AFM was used to investigate the morphological, nanostructural, cytoskeletal, and mechanical properties of red blood cells (RBCs). Based on experimental data, a set of important biomarkers determining the status of blood cells have been identified.

Topological Relationships Cytoskeleton-Membrane Nanosurface-Morphology as a Basic Mechanism of Total Disorders of RBC Structures.

The state of red blood cells (RBCs) and their functional possibilities depend on the structural organization of the membranes. Cell morphology and membrane nanostructure are compositionally and functionally related to the cytoskeleton network. In this work, the influence of agents (hemin, endogenous oxidation during storage of packed RBCs, ultraviolet (UV) radiation, temperature, and potential of hydrogen (pH) changes) on the relationships between cytoskeleton destruction, membrane nanostructure, and RBC morphology was observed by atomic force microscope. It was shown that the influence of factors of a physical and biochemical nature causes structural rearrangements in RBCs at all levels of organization, forming a unified mechanism of disturbances in relationships "cytoskeleton-membrane nanosurface-cell morphology". Filament ruptures and, consequently, large cytoskeleton pores appeared. The pores caused membrane topological defects in the form of separate grain domains. Increasing loading doses led to an increase in the number of large cytoskeleton pores and defects and their fusion at the membrane nanosurfaces. This caused the changes in RBC morphology. Our results can be used in molecular cell biology, membrane biophysics, and in fundamental and practical medicine.

Two-step process of cytoskeletal structural damage during long-term storage of packed red blood cells.

BACKGROUND: Storage of packed red blood cells (PRBC) for 42 days causes morphological, structural, and functional changes in the red cells. To assess the quality of stored PRBC, it is important to evaluate the main components of the product. The aim of this study was to evaluate the kinetics of the structural transformations in the cytoskeleton of red cells during long-term storage (up to 42 days). MATERIALS AND METHODS: Bags of PRBC were stored with CPD/SAGM solution at +4 °C. Cytoskeletal parameters were measured on days 3, 12, 19, 21, 24, 28, 35, and 42 of storage to determine their changes. Atomic force microscopy was used to obtain images and analyse the parameters of the cytoskeletal network. As the storage time increased, a general PRBC test was performed. Membrane fixatives were not used at any stage of the preparation of the specimens for cytoskeletal imaging. RESULTS: When PRBC were stored for 42 days, the main changes to the cytoskeletal mesh included rupture of filaments, merger of small pores into larger ones, a decrease of the number of pores, thickening of filaments, and an increase of membrane stiffness. A process of irreversible changes to the cytoskeleton started on days 19-21. A kinetic model of changes in the parameters of the cytoskeletal mesh with time of PRBC storage was created. DISCUSSION: Two stages of impairment in cytoskeletal elements were found: rupture of filaments and clustering of protein components. The typical time of development and specifics of these stages are discussed. The consequences of the altered configuration of the cytoskeleton are also discussed. Destruction of the red cell cytoskeleton can have a negative effect on the efficacy of blood transfusion and increase the risk of post-transfusion complications. Our findings can be used in clinical medicine to evaluate the quality of PRBC for blood transfusion as well as for studies of the molecular organisation of red cells undergoing various types of physical and chemical treatment.

Assessment of carboxyhemoglobin content in the blood with high accuracy: wavelength range optimization for nonlinear curve fitting of optical spectra.

The impact of carbon monoxide (CO) gas on the human organism is very dangerous. The affinity of CO to hemoglobin is considerably higher than that of oxygen. Thus, the interaction of CO with the blood results in a higher content of carboxyhemoglobin (HbCO) in red blood cells (RBCs) and correspondingly in tissue hypoxia. The disruption in the organism depends on the HbCO content in the blood. To assess any complications in the body at a given moment due to CO exposure and predict future consequences, it is necessary to measure the dynamics of hemoglobin derivative concentrations simultaneously. However, measuring HbCO and other derivatives in RBCs without hemolysis accurately is complicated due to the strong intercollinearity between the molar absorptivities of hemoglobin derivatives and superposition of absorption and scattering spectra. In the present study, to quantitatively assess the contents of the hemoglobin derivatives in the blood after exposure to CO, improved accuracy is achieved by optimizing the wavelength range used for the nonlinear curve fitting of optical spectra. Experimental spectra were measured in the wavelength range Δ λ = 500 - 700 nm . For each experimental curve, it was established the value of optimal interval Δ λ o p t for which the correlation coefficient between experimental data and corresponding points of the theoretical fitting curve was the maximum in the wavelength range Δ λ t y p = 535 - 580 n m , which contains the typical absorption peaks for H b O 2 , H b , and H b C O . The concentrations obtained based on such fitting curves were considered to be highly accurate. The quantitative assessment enabled the determination of the H b C O nonlinear increase with the time of CO exposure in the in vitro experiment and the study of the dynamics of hemoglobin derivative transformations during blood incubation.

Conformational Distortions of the Red Blood Cell Spectrin Matrix Nanostructure in Response to Temperature Changes In Vitro.

The spectrin matrix is a structural element of red blood cells (RBCs). As such, it affects RBC morphology, membrane deformability, nanostructure, stiffness, and, ultimately, the rheological properties of blood. However, little is known about how temperature affects the spectrin matrix. In this study, the nanostructure of the spectrin network was recorded by atomic force microscopy. We describe how the nanostructure of the RBC spectrin matrix changes from a regular network to a chaotic pattern following an increase in temperature from 20 to 50°C. At 20-37°Ð¡, the spectrin network formed a regular structure with dimensions of typically 150 ± 60 nm. At 42-43°Ð¡, 83% of the spectrin network assumed an irregular structure. Finally, at 49-50°Ð¡ the chaotic pattern was observed, and no quantitative estimates of the spectrin structure's parameters could be made. These results can be useful for biophysical studies on the destruction of the spectrin network under pathological conditions, as well as for investigating cell morphology and blood rheology in different diseases.