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- Effect of antioxidant and optimal antimicrobial mixtures of carvacrol, grape seed extract and chitosan on different spoilage microorganisms and their application as coatings on different food matricesPublication . Rubilar, Javiera F.; Cruz, R. M. S.; Khmelinskii, Igor; Vieira, M. M. C.There is growing interest in the use of natural agents with antimicrobial (AM) and antioxidant (AOX) properties. Optimization of the AM capacity for mixtures containing carvacrol, grape seed extract (GSE) and chitosan, against gram-negative (Pseudomonas aeruginosa), gram-positive bacteria (Staphylococcus aureus, Listeria innocua and Enterococcus faecalis) and yeast (Saccharomyces cerevisiae) at 106 cfu mL−1 was studied. To observe the synergistic or antagonistic effect and find optimal combinations between the three agents, a simplex centroid mixture design was run for each microorganism, combining carvacrol (0-300 ppm, X1)X, GSE (0-2000 ppm, X2) and chitosan (0-2% w/v, X3). Results of the response surface analysis showed several synergistic effects for all microorganisms. Combinations of 60 ppm-400 ppm-1.2% w/v (carvacrol-GSE-chitosan; optimal AM combination 1, OAMC-1); 9.6 ppm-684 ppm-1.25% w/v (OAMC-2); 90 ppm-160 ppm-1.24% w/v (OAMC-3) were found to be the optimal mixtures for all microorganisms. Radical scavenging activity (RSA) of the same agents was then compared with a standard AOX (butylated hydroxytoluene; BHT) at different concentrations (25, 50 and 100 ppm; as well as the optimal AM concentrations) by the 1,1-diphenyl-2- picrylhydrazyl (DPPH) method. RSA increased in the following order: chitosan< carvacrol< BHT< GSE and for the OAMC: OAMC-2< OAMC-1< OAMC-3. The best RSA (OAMC-3) was applied as a coating in two different food matrices (strawberries and salmon). For strawberries, P. aeruginosa was more sensitive to the action of OAMC-3 than S. cerevisiae. For salmon, S. aureus was more resistant to the action of OAMC-3 than E. faecalis and L. innocua.
- Mathematical modeling of gallic acid release from chitosan films with grape seed extract and carvacrolPublication . Rubilar, Javiera F.; Cruz, Rui; Zuñiga, Rommy N.; Khmelinskii, Igor; Vieira, M. M. C.Controlled release of antimicrobial and antioxidant compounds from packaging films is of utmost importance for extending the shelf-life of perishable foods. This study focused on the mathematical modeling of gallic acid release into an aqueous medium from three chitosan films, formulated with grape seed extract (GSE) and carvacrol. We quantified the release by HPLC technique during 30days at three temperatures (5, 25 and 45°C). The diffusion coefficients, varying with temperature according to an Arrhenius-type relationship, and the respective activation energies for Film-1 and Film-2 were, respectively [Formula: see text] m2s-1 and [Formula: see text] m2s-1, Ea1=58kJmol-1 and Ea2=60kJmol-1 as obtained from the Fickian fit. The low concentrations of gallic acid released by Film-3 could not be detected by HPLC, therefore the respective diffusion coefficient was not estimated. This study will help with the development and optimization of active packaging (AP) films aiming at improved food preservation and shelf-life extension.
- Focusing effects of ballistic transverse-quantized excitons in metal nanofilmsPublication . Makarov, Vladimir; Khmelinskii, IgorNew type of behavior of transverse-quantized excitons was discovered in metal nanofilms. These excitons demonstrated ballistic properties, propagating along nanofilms in straight lines along centimeter distances, reflecting at the film patch boundaries, and refracting at the boundary separating films of different materials but with the same exciton energy. Exciton reflection and focusing was explored in elliptically shaped metal film patches, along with exciton refraction on the boundary line separating two different film patches with compatible transverse-confinementgenerated electronic structures. These transverse-quantized excitons interact with phonons very weakly, prohibited by symmetry selection rules. The latter statement was confirmed in timeresolved experiments. Weak interactions explain rectilinear trajectories and long lifetimes of excitons in thin metal films.
- Front-face fluorescence spectroscopy and chemometrics for quality control of cold-pressed rapeseed oil during storagePublication . Sikorska, Ewa; Wójcicki, Krzysztof; Kozak, Wojciech; Gliszczyńska-Świgło, Anna; Khmelinskii, Igor; Górecki, Tomasz; Caponio, Francesco; Paradiso, Vito M.; Summo, Carmine; Pasqualone, AntonellaThe aim of this study was to test the usability of fluorescence spectroscopy to evaluate the stability of cold-pressed rapeseed oil during storage. Freshly-pressed rapeseed oil was stored in colorless and green glass bottles exposed to light, and in darkness for a period of 6 months. The quality deterioration of oils was evaluated on the basis of several chemical parameters (peroxide value, acid value, K232 and K270, polar compounds, tocopherols, carotenoids, pheophytins, oxygen concentration) and fluorescence. Parallel factor analysis (PARAFAC) of oil excitation-emission matrices revealed the presence of four fluorophores that showed different evolution throughout the storage period. The fluorescence study provided direct information about tocopherol and pheophytin degradation and revealed formation of a new fluorescent product. Principal component analysis (PCA) performed on analytical and fluorescence data showed that oxidation was more advanced in samples exposed to light due to the photo-induced processes; only a very minor effect of the bottle color was observed. Multiple linear regression (MLR) and partial least squares regression (PLSR) on the PARAFAC scores revealed a quantitative relationship between fluorescence and some of the chemical parameters.
- Reversible and irreversible mitochondrial swelling in vitroPublication . Khmelinskii, Igor; Makarov, VladimirMitochondrial activity as regards ATP production strongly depends on mitochondrial swelling (MS) mode. Therefore, this work analyzes reversible and irreversible MS using a detailed biophysical model. The reported model includes mechanical properties of the inner mitochondrial membrane (IMM). The model describes MS dynamics for spherically symmetric, axisymmetric ellipsoidal and general ellipsoidal mitochondria. Mechanical stretching properties of the IMM were described by a second-rank rigidity tensor. The tensor components were estimated by fitting to the earlier reported results of in vitro experiments. The IMM rigidity constant of ca. 0.008 dyn/nm was obtained for linear deformations. The model also included membrane bending effects, which were small compared to those of membrane stretching. The model was also tested by simulation of the earlier reported experimental data and of the system dynamics at different initial conditions, predicting the system behavior. The transition criteria from reversible to irreversible swelling were determined and tested. The presently developed model is applicable directly to the analysis of in vitro experimental data, while additional improvements are necessary before it could be used to describe mitochondrial swelling in vivo. The reported theoretical model also provides an idea of physically consistent mechanism for the permeability transport pore (PTP) opening, which depends on the IMM stretching stress. In the current study, this idea is discussed briefly, but a detailed theoretical analysis of these ideas will be performed later. The currently developed model provides new understanding of the detailed MS mechanism and of the conditions for the transition between reversible and irreversible MS modes. On the other hand, the current model provides useful mathematical tools, that may be successfully used in mitochondrial biophysics research, and also in other applications, predicting the behavior of mitochondria in different conditions of the surrounding media in vitro or cellular cyto(sarco)plasm in vivo. These mathematical tools are based on real biophysical processes occurring in mitochondria. Thus, we note a significant progress in the theoretical approach, which may be used in real biological systems, compared to the earlier reported models. Significance of this study derives from inclusion of IMM mechanical properties, which directly impact the reversible and irreversible mitochondrial swelling dynamics. Reversible swelling corresponds to reversible IMM deformations, while irreversible swelling corresponds to irreversible deformations, with eventual membrane disruption. The IMM mechanical properties are directly dependent on the membrane biochemical composition and structure. The IMM deformationas are induced by osmotic pressure created by the ionic/neutral solute imbalance between the mitochondrial matrix media and the bulk solution in vitro, or cyto(sarco)plasm in vivo. The novelty of the reported model is in the biophysical mechanism detailing ionic and neutral solute transport for a large number of solutes, which were not taken into account in the earlier reported biophysical models of MS. Therefore, the reported model allows understanding response of mitochondria to the changes of initial concentration(s) of any of the solute(s) included in the model. Note that the values of all of the model parameters and kinetic constants have been estimated and the resulting complete model may be used for quantitative analysis of mitochondrial swelling dynamics in conditions of real in vitro experiments.
- Science in the times of Covid. An alternative hypothesisPublication . Stallinga, Peter; Khmelinskii, Igor; Woodcock, LeslieScience consists of testing hypothesis. However, this Scientific Method is used ever more scarcely, and is replaced by general research to help making society a better place. In the current work we analyze how the approach of a solution to the sanitary problem caused by the SARS-CoV-2 virus has been done in a non-scientific way leading to erroneous conclusions. Moreover, we form an alternative hypothesis that has withstood our own attempts at debunking. We conclude that the pandemic is caused by misdiagnosis of other respiratory illnesses and a runaway-testing-scenario.
- Analysis of quantum coherence in biologyPublication . Khmelinskii, Igor; Makarov, Vladimir, IWe reviewed the tools of quantum physics used in modeling of quantum coherence (QC) effects in different systems, including biological systems, which behave as quantum objects in some of their degrees of freedom. In particular, we considered the usage of the effective Hamiltonian (EH), Green's function (GF) and density matrix (DM) methods in the analysis of QC, focusing on QC in biological systems. We discussed the two main mechanisms of loss of quantum state coherence: (i) dephasing of the originally prepared coherent wave package and (ii) population relaxation in the same wave package. Dephasing does not affect the quantum state population, e.g. as in spin-spin relaxation, where dephasing is described by the tau(2) relaxation time. On the other hand, the state population relaxation of the spin wavepackage is attributed to spin-lattice relaxation and is described by the tau(1) relaxation time. Presently we discussed EH and GF formalisms in terms of the complex energy, dependent on intra- and intersystem interactions that induce state population relaxation. We provided a detailed analysis of these approaches for the exciton relaxation dynamics in a glycine polypeptide chain. The same phenomena were described in the DM formalism using the relaxation matrix. We discussed QC in different biological systems, showing that QC is conserved when the interactions of the coherent wavepackage with other degrees of freedom are weak, as otherwise population relaxation causes loss of QC. We believe that our results will be useful for the researchers in the area of quantum biology.
- Computational modeling of In vitro swelling of mitochondria: A biophysical approachPublication . Makarov, Vladimir I.; Khmelinskii, Igor; Javadov, SabzaliSwelling of mitochondria plays an important role in the pathogenesis of human diseases by stimulating mitochondria-mediated cell death through apoptosis, necrosis, and autophagy. Changes in the permeability of the inner mitochondrial membrane (IMM) of ions and other substances induce an increase in the colloid osmotic pressure, leading to matrix swelling. Modeling of mitochondrial swelling is important for simulation and prediction of in vivo events in the cell during oxidative and energy stress. In the present study, we developed a computational model that describes the mechanism of mitochondrial swelling based on osmosis, the rigidity of the IMM, and dynamics of ionic/neutral species. The model describes a new biophysical approach to swelling dynamics, where osmotic pressure created in the matrix is compensated for by the rigidity of the IMM, i.e., osmotic pressure induces membrane deformation, which compensates for the osmotic pressure effect. Thus, the effect is linear and reversible at small membrane deformations, allowing the membrane to restore its normal form. On the other hand, the membrane rigidity drops to zero at large deformations, and the swelling becomes irreversible. As a result, an increased number of dysfunctional mitochondria can activate mitophagy and initiate cell death. Numerical modeling analysis produced results that reasonably describe the experimental data reported earlier.
- Screening of antioxidant properties of the apple juice using the front-face synchronous fluorescence and chemometricsPublication . Wlodarska, Katarzyna; Pawlak-Lemanska, Katarzyna; Khmelinskii, Igor; Sikorska, EwaFluorescence spectroscopy is gaining increasing attention in food analysis due to its higher sensitivity and selectivity as compared to other spectroscopic techniques. Synchronous scanning fluorescence technique is particularly useful in studies of multi-fluorophoric food samples, providing a further improvement of selectivity by reduction in the spectral overlapping and suppressing light-scattering interferences. Presently, we study the feasibility of the prediction of the total phenolics, flavonoids, and antioxidant capacity using front-face synchronous fluorescence spectra of apple juices. Commercial apple juices from different product ranges were studied. Principal component analysis (PCA) applied to the unfolded synchronous fluorescence spectra was used to compare the fluorescence of the entire sample set. The regression analysis was performed using partial least squares (PLS1 and PLS2) methods on the unfolded total synchronous and on the single-offset synchronous fluorescence spectra. The best calibration models for all of the studied parameters were obtained using the PLS1 method for the single-offset synchronous spectra. The models for the prediction of the total flavonoid content had the best performance; the optimal model was obtained for the analysis of the synchronous fluorescence spectra at Delta lambda = 110 nm (R (2) = 0.870, residual predictive deviation (RPD) = 2.7). The optimal calibration models for the prediction of the total phenolic content (Delta lambda = 80 nm, R (2) = 0.766, RPD = 2.0) and the total antioxidant capacity (Delta lambda = 70 nm, R (2) = 0.787, RPD = 2.1) had only an approximate predictive ability. These results demonstrate that synchronous fluorescence could be a useful tool in fast semi-quantitative screening for the antioxidant properties of the apple juices.
- Quantum mechanism of light energy propagation through an avian retinaPublication . Zueva, Lidia; Golubeva, Tatiana; Korneeva, Elena; Resto, Oscar; Inyushin, Mikhail; Khmelinskii, Igor; Makarov, VladimirTaking into account the ultrastructure of the Pied Flycatcher foveal retina reported earlier and the earlier reported properties of Muller cell (MC) intermediate filaments (IFs) isolated from vertebrate retina, we proposed a quantum mechanism (QM) of light energy transfer from the inner limiting membrane level to visual pigments in the photoreceptor cells. This mechanism involves electronic excitation energy transfer in a donor-acceptor system, with the IFs excited by photons acting as energy donors, and visual pigments in the photoreceptor cells acting as energy acceptors. It was shown earlier that IFs with diameter 10 nm and length 117 mu m isolated from vertebrate eye retina demonstrate properties of light energy guide, where exciton propagates along such IFs from MC endfeet area to photoreceptor cell area. The energy is mostly transferred via the contact exchange quantum mechanism. Our estimates demonstrate that energy transfer efficiencies in such systems may exceed 80-90%. Thus, the presently developed quantum mechanism of light energy transfer in the inverted retina complements the generally accepted classic optical mechanism and the mechanism whereby Muller cells transmit light like optical fibers. The proposed QM of light energy transfer in the inverted retina explains the high image contrast achieved in photopic conditions by an avian eye, being probably also active in other vertebrates.