Targeting Oxidative Stress and Inflammation in the Eye: Insights from a New Model of Experimental Autoimmune Uveitis.

Autoimmune uveitis is a relapsing blind-causing ocular condition with complex pathogenesis that is not completely understood. There is a high demand for accurate animal models of experimental autoimmune uveitis (EAU) suitable for elucidating the molecular mechanisms of the disease and testing new therapeutic approaches. Here, we demonstrated that photoreceptor Ca/Zn-sensor protein recoverin is a uveoretinal antigen in albino rabbits provoking typical autoimmune chorioretinitis 2-4 weeks after immunization.

Expression, Intracellular Localization, and Maturation of Cysteine Cathepsins in Renal Embryonic and Cancer Cell Lines.

Cysteine cathepsins play an important role in tumor development and metastasis. The expression of these enzymes is often increased in many types of tumor cells. Cysteine cathepsins contribute to carcinogenesis through a number of mechanisms, including proteolysis of extracellular matrix and signaling molecules on the cell surface, as well as degradation of transcription factors and disruption of signaling cascades in the cell nucleus.

Redox Regulation of Signaling Complex between Caveolin-1 and Neuronal Calcium Sensor Recoverin.

Caveolin-1 is a cholesterol-binding scaffold protein, which is localized in detergent-resistant membrane (DRM) rafts and interacts with components of signal transduction systems, including visual cascade. Among these components are neuronal calcium sensors (NCSs), some of which are redox-sensitive proteins that respond to calcium signals by modulating the activity of multiple intracellular targets. Here, we report that the formation of the caveolin-1 complex with recoverin, a photoreceptor NCS serving as the membrane-binding regulator of rhodopsin kinase (GRK1), is a redox-dependent process.

Possible risks for the functioning of cyclic processes in the experimental model of a closed ecosystem.

The purpose of the present study is to consider a number of possible risks that may emerge when processed human wastes are involved into mass exchange processes as fertilizers for plants cultivated in the experimental model of the closed ecosystem (CEEM). The problems relating to the disruption of cycling processes in closed ecosystems can be tentatively divided into two groups: the problems that can be rather easily overcome and the chronic problems. Addition of plant inedible biomass to the soil-like substrate (SLS) can result in a decrease in plant productivity because of allelopathic interactions and enhanced growth of microorganisms.

Alterations in proteome of human sclera associated with primary open-angle glaucoma involve proteins participating in regulation of the extracellular matrix.

Purpose: Primary open-angle glaucoma (POAG) is a common ocular disease, associated with abnormalities in aqueous humor circulation and an increase in intraocular pressure (IOP), leading to progressive optical neuropathy and loss of vision. POAG pathogenesis includes alterations of the structural properties of the sclera, especially in the optic nerve head area, contributing to the degeneration of the retinal ganglion cells. Abnormal sclera biomechanics hinder adequate compensation of IOP fluctuations, thus aggravating POAG progression.

A Novel Approach to Bacterial Expression and Purification of Myristoylated Forms of Neuronal Calcium Sensor Proteins.

N-terminal myristoylation is a common co-and post-translational modification of numerous eukaryotic and viral proteins, which affects their interaction with lipids and partner proteins, thereby modulating various cellular processes. Among those are neuronal calcium sensor (NCS) proteins, mediating transduction of calcium signals in a wide range of regulatory cascades, including reception, neurotransmission, neuronal growth and survival. The details of NCSs functioning are of special interest due to their involvement in the progression of ophthalmological and neurodegenerative diseases and their role in cancer.

Deep Physical-Chemical Purification of Gas Medium in Artificial Ecosystems.

The results of experiments on application of a newly developed facility for oxidation of volatile organic compounds on a platinum catalyst are presented. The feasibility of using this method in artificial ecosystems as a whole and in mass exchange of closed biological-technical life support systems in particular is shown. The possibility of deep purification of gas emitted from the reactor of physical-chemical processing of organic wastes is demonstrated.

Mechanisms and Treatment of Light-Induced Retinal Degeneration-Associated Inflammation: Insights from Biochemical Profiling of the Aqueous Humor.

Ocular inflammation contributes to the pathogenesis of blind-causing retinal degenerative diseases, such as age-related macular degeneration (AMD) or photic maculopathy. Here, we report on inflammatory mechanisms that are associated with retinal degeneration induced by bright visible light, which were revealed while using a rabbit model. Histologically and electrophysiologically noticeable degeneration of the retina is preceded and accompanied by oxidative stress and inflammation, as evidenced by granulocyte infiltration and edema in this tissue, as well as the upregulation of total protein, pro-inflammatory cytokines, and oxidative stress markers in aqueous humor (AH).

Membrane Binding of Neuronal Calcium Sensor-1: Highly Specific Interaction with Phosphatidylinositol-3-Phosphate.

Neuronal calcium sensors are a family of N-terminally myristoylated membrane-binding proteins possessing a different intracellular localization and thereby targeting unique signaling partner(s). Apart from the myristoyl group, the membrane attachment of these proteins may be modulated by their N-terminal positively charged residues responsible for specific recognition of the membrane components. Here, we examined the interaction of neuronal calcium sensor-1 (NCS-1) with natural membranes of different lipid composition as well as individual phospholipids in form of multilamellar liposomes or immobilized monolayers and characterized the role of myristoyl group and N-terminal lysine residues in membrane binding and phospholipid preference of the protein.

Novel applications of modification of thiol enzymes and redox-regulated proteins using S-methyl methanethiosulfonate (MMTS).

S-Methyl methanethiosulfonate (MMTS) is used in experimental biochemistry for alkylating thiol groups of protein cysteines. Its applications include mainly trapping of natural thiol-disulfide states of redox-sensitive proteins and proteins which have undergone S-nitrosylation. The reagent can also be employed as an inhibitor of enzymatic activity, since nucleophilic cysteine thiolates are commonly present at active sites of various enzymes.

[Investigation of the iron-binding capacity of the bovine lactoferrin].

Unlabelled: In this work, studies were carried out to obtain and determine the iron-binding ability of lactoferrin isolated from milk of Holstein-Friesian (black-and-white) breed of cows, which is the main stock of the Russian cattle herd (CH). Aim of the study was to obtain lactoferrin and determine its iron-binding capacity for substantiating the raw material resources of its industrial production as an easily digestible source of ferrous iron for the production of dietary supplements and/or specialized foods.

Material And Methods: To optimize the production of lactoferrin in the conditions of dairy enterprises, we used a method of lactoferrin isolation from cow's milk in its own modification, which consisted in the degreasing of whole milk by centrifugation and double cation-exchange chromatography with successive application of the following sorbents: CM-cellulose (CM-52) and Macro-Prep High Q Support.

Incorporation of mineralized human waste and fish waste as a source of higher plant mineral nutrition in the BTLSS mass exchange.

The present study deals with the development of the principles and conditions of fish waste mineralization using the method of wet combustion with hydrogen peroxide in alternating electromagnetic field and describes testing mineralized human waste and fish waste as sources of nutrients for plants in the biotechnical human life support system (BTLSS). The study shows that mineralization of fish waste in the wet combustion reactor should be performed in the presence of readily oxidized organic matter, represented by human waste, as an activator of oxidation. Re-mineralization of the sediment in the mixture of hydrogen peroxide and nitric acid in the wet combustion reactor converts mineral elements bound in the sediment into the form available to plants.

Light-Induced Thiol Oxidation of Recoverin Affects Rhodopsin Desensitization.

The excessive light illumination of mammalian retina is known to induce oxidative stress and photoreceptor cell death linked to progression of age-related macular degeneration. The photochemical damage of photoreceptors is suggested to occur via two apoptotic pathways that involve either excessive rhodopsin activation or constitutive phototransduction, depending on the light intensity. Both pathways are dramatically activated in the absence of rhodopsin desensitization by GRK1.

Suppression of Light-Induced Oxidative Stress in the Retina by Mitochondria-Targeted Antioxidant.

Light-induced oxidation of lipids and proteins provokes retinal injuries and results in progression of degenerative retinal diseases, such as, for instance, iatrogenic photic maculopathies. Having accumulated over years retinal injuries contribute to development of age-related macular degeneration (AMD). Antioxidant treatment is regarded as a promising approach to protecting the retina from light damage and AMD.

A small closed ecosystem with an estimated portion of human metabolism.

The study describes a small closed ecosystem used to test technologies to be further employed in full-scale manned closed ecosystems. The experimental ecosystem is designed to use a certain portion of human metabolism, which is included in the gas, water, and organic waste loops of the system. In this experimental ecosystem, gas and water loops are fully closed, and the model enables processing of human waste and plant inedible biomass.

Photoreceptor calcium sensor proteins in detergent-resistant membrane rafts are regulated via binding to caveolin-1.

Rod cell membranes contain cholesterol-rich detergent-resistant membrane (DRM) rafts, which accumulate visual cascade proteins as well as proteins involved in regulation of phototransduction such as rhodopsin kinase and guanylate cyclases. Caveolin-1 is the major integral component of DRMs, possessing scaffolding and regulatory activities towards various signaling proteins. In this study, photoreceptor Ca-binding proteins recoverin, NCS1, GCAP1, and GCAP2, belonging to neuronal calcium sensor (NCS) family, were recognized as novel caveolin-1 interacting partners.

[Quantitative evaluation of uveoscleral outflow for optimization of glaucoma treatment].

Unlabelled: The leading role in glaucoma treatment is now played by prostaglandin analogues (PGAs), whose point of application is the uveoscleral outflow of aqueous humor. Quantitative evaluation of the latter is, however, a problem yet unsolved.

Aim: To assess the clinical applicability of a new method for quantitative evaluation of the uveoscleral outflow in human eyes, which is meant to help with optimization of glaucoma therapy.

Light-induced disulfide dimerization of recoverin under ex vivo and in vivo conditions.

Despite vast knowledge of the molecular mechanisms underlying photochemical damage of photoreceptors, linked to progression of age-related macular degeneration, information on specific protein targets of the light-induced oxidative stress is scarce. Here, we demonstrate that prolonged intense illumination (halogen bulb, 1500 lx, 1-5 h) of mammalian eyes under ex vivo (cow) or in vivo (rabbit) conditions induces disulfide dimerization of recoverin, a Ca(2+)-dependent inhibitor of rhodopsin kinase. Western blotting and mass spectrometry analysis of retinal extracts reveals illumination time-dependent accumulation of disulfide homodimers of recoverin and its higher order disulfide cross-linked species, including a minor fraction of mixed disulfides with intracellular proteins (tubulins, etc.

[Use of the ion-exchange substrate to optimize mineral nutrition of plants within a bio-engineering life support system with a high level of closure].

Purpose of the work was to test manageability of nutrient solutions containing mineralized human exometabolites by using an ion-exchange substrate (IES) for cultivating wheat in a bio-engineering life support system with a high level of closure. Object of the investigation was wheat Triticum aestivum L. (Lysovsky cv.

Synergetic effect of recoverin and calmodulin on regulation of rhodopsin kinase.

Phosphorylation of photoactivated rhodopsin by rhodopsin kinase (RK or GRK1), a first step of the phototransduction cascade turnoff, is under the control of Ca(2+)/recoverin. Here, we demonstrate that calmodulin, a ubiquitous Ca(2+)-sensor, can inhibit RK, though less effectively than recoverin does. We have utilized the surface plasmon resonance technology to map the calmodulin binding site in the RK molecule.

[Antioxidant activity of cationic whey protein isolate].

The process of lipid peroxidation (LPO) in biological membranes of cells is carried out by free radical mechanism, a feature of which is the interaction of radicals with other molecules. In this work we investigated the antioxidant activity of cationic whey protein isolate, obtained by the cation-exchange chromatography on KM-cellulose from raw cow's milk, in vitro and in vivo. In biological liquids, which are milk, blood serum, fetal fluids, contains a complex of biologically active substances with a unique multifunctional properties, and which are carrying out a protective, antimicrobial, regenerating, antioxidant, immunomodulatory, regulatory and others functions.

Amino acid sequences of two immune-dominant epitopes of recoverin are involved in Ca2+/recoverin-dependent inhibition of phosphorylation of rhodopsin.

Antibodies AB(60-72) and AB(80-92) against two immune-dominant epitopes of photoreceptor Ca(2+)-binding protein recoverin, 60-DPKAYAQHVFRSF-72 and 80-LDFKEYVIALHMT-92, which can be exposed in a Ca(2+)-dependent manner, were obtained. The presence of AB(60-72) or AB(80-92) results in a slight increase in Ca(2+)-affinity of recoverin and does not affect significantly a Ca(2+)-myristoyl switch mechanism of the protein. However in the presence of AB(60-72) or AB(80-92) recoverin loses its ability to interact with rhodopsin kinase and consequently to perform a function of Ca(2+)-sensitive inhibitor of rhodopsin phosphorylation in photoreceptor cells.

[Possible ways to include human exometabolites in the turnover of biological life support system].

Purpose of the investigation was to study feasibility to include solid and liquid human discharges in the turnover of bioregenerative life support systems for physical/chemical and biological treatment. Human exometabolites were mineralized by way of wet process in hydrogen peroxide in alternating electromagnetic field. The resultant solutions were used to water plants cultivated on a soil-like substrate or haydite.