Viruses of genus Carlavirus encode a small cysteine-rich protein (CRP) of unknown function. To investigate the role of CRP of carlavirus chrysanthemum virus B (CVB), a recombinant potato virus X (PVX) genome was constructed, which carried the CVB CRP gene. Expression of CVB CRP in the PVX genetic background drastically changed the PVX symptom phenotype in N. benthamiana. Instead of symptomless infection and mild mosaic, which are characteristic of PVX in this plant host, the recombinant virus expressing CVB CRP induced formation of necrotic local lesions on inoculated leaves and necrosis of the apical leaves. In N. tabacum, the infection pattern depended on the host genotype: the recombinant PVX was able to spread systemically only in N gene-carrying plants. In agroinfiltration-mediated transient expression assay, CVB CRP did not exhibit the properties of avirulence factor in N. benthamiana and was unable to suppress post-transcriptional gene silencing. Thus, CVB CRP is the viral pathogenicity determinant controlling the virus interaction with plant hosts in a manner which depends on plant defense mediated by resistance genes such as the N gene.
We studied the metabolic changes induced by gaseous nitric oxide in whole blood samples in vitro. Blood samples were collected from healthy donors (Nizhny Novgorod station of blood transfusion). We carried out the direct bubbling of blood samples (n = 14) with gaseous flow with NO in a special appliance. We modeled standard conditions using the apparatus "Plazon" (concentration NO 800 mcg/l). Middle power of gas flow was used. The blood sparging time was 2 min, and exposition time lasted 3 min. Every blood sample volume was 5 ml. All the parameters were controlled before and after blood processing with NO. We tested lactate dehydrogenase activity in direct and reverse reactions spectrometrically by G. A. Kochetov's method. Aldehyde dehydrogenase activity was examined by B. M. Kershnhots's and E. V. Serkina's methods, superoxide dismutase—by T. V. Sirota's technology. Total protein level was examined by modified Louri's method. The concentration of lactate was tested with the automatic analyzer "SuperGL Ambulance". The indices of acidbase balance and blood gases partial pressure were estimated with special analyzer "ABL-77". Additional control of energy metabolism changes was accomplished with derivative parameters, such as coefficient of energy reaction balance and coefficient of substrate provision. Different changes of blood physical and chemical parameters are induced by NO-processing which was fixed in our experiments. There is an inhibition of erythrocytes energy metabolism, decreasing of plasma antioxidant reserves, moderate ionic disorders and of acid-base misbalance in blood samples in vitro. Besides, according to the indirect signs, the used regimen of NO-processing mainly affected erythrocytes, and stipulated methemoglobin formation. These data testify that the used dose of gaseous nitric oxide is too high for investigated human blood. In our opinion, registered negative effects of free NO may be eliminated by bound nitric oxide use (first of all in its natural form—dinitrosyl-iron complexes).
The formulation of the transdermal patch with fulvic acid (FA) on an emulsion basis using pluronic Kolliphor® p237 as a permeability enhancer was developed and studied for anti-inflammatory properties. FA was isolated from the peat in the Nizhny Novgorod region of Russia and characterized as a potential active pharmaceutical ingredient. In vitro studies of the release of FA from the transdermal patch, as well as the FA penetration through the acetyl cellulose membrane using the Franz diffusion cell, showed its high efficiency (56% and 90%, respectively, in 8 h). In the in vivo experiment, qualitative and quantitative features of the rat knee caused by complete Freund’s adjuvant-induced arthritis (morphological changes, the FA influence on the biochemical indexes) were studied. The inflammatory process that developed within 15 days was accompanied by the activation of antioxidant oxidoreductase enzymes (by 50–70%), the increase in the cross-sectional diameter of the cartilage, and the increase in the values of marker indicators of the process of rheumatoid arthritis. Within 7 days of treatment, under the influence of FA, the values of ESR, RF, leukocytes, C-reactive protein, as well as the biochemical parameters characterizing oxidative stress (SOD, catalase, glutathione reductase, LDH, G6PD) normalized, and the edema reduced. These results may be useful for arthritis treatment using the transdermal patch with FA.
To make multiparametric analysis of blood metabolic profile in early period of burn disease.We tested blood samples of 15 healthy adults (control group) and 60 patients with thermal trauma (main group--II-4IIIA, B degree of burn, more then 15 bsp). Parameters of lipid metabolism, level of glucose, lactate, malonic dialdehyde and some enzymes in blood plasma and erythrocytes were estimated.In early period of burn disease we fixed the clear metabolic disorders, including tissues hypoxia, activation of plasma transaminases and oxidoreductases, inhibition of detoxication system, induction of oxidative stress. Connection of metabolic changes, associated with burn disease, was registered. It supported by numerous correlations between studied parameters, formed from first day after trauma.Our data expand the knowledge about mating metabolic changes of catalytic activity of blood enzymes, forming in early period of burn disease (system metabolic disadaptation), and diagnostic value of some blood biochemical parameters in estimation of burned patient metabolism.
Background. Ozonation products of unsaturated fatty acids in fats and oils – 1,2,4-trioxolanes – which contribute to the normalization of oxidative and energy metabolism in various diseases, may be a promising substance for the creation of new combined anti-burn drugs. The aim. To study the effect of 1,2,4-trioxolanes in a pharmaceutical composition with betulin, which exhibits wound-healing properties, on indicators of oxidative stress and on energy metabolism in the treatment of burns in an experiment onrats. Methods. The experiments were carried out on 50 male Wistar rats with seconddegree skin burns. We assessed the activity of glutathione reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, aldehyde dehydrogenase, superoxide dismutase and catalase, as well as the level of lipid peroxidation indicators. Results. The properties of 1,2,4-trioxolanes were studied and a pharmaceutical composition of 1,2,4-trioxolanes with betulin from birch bark in fish oil was developed. The effect of this composition on oxidative and energy metabolism in rats during the treatment of burns was studied. The activation of NADP/H and NAD/H dependent enzymes (glutathione reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, aldehyde dehydrogenase), as well as superoxide dismutase and catalase, and the normalization of lipid peroxidation parameters under the influence of the studied composition in the treatment of burns were revealed. The effect of the composition on the energy metabolism of erythrocytes was noted, which was assessed by changes in the ratio of lactate dehydrogenase activity in the reverse and direct reactions. Morphohistological studies showed that using the composition under normal conditions and in case of a burn had no effect on the structure of the stomach wall, and also improved the condition of the burn wound on the day 10 compared to the wound condition in animals without treatment. Conclusion. The proposed composition of 1,2,4-trioxolanes and betulin in fish oil, which allows normalizing oxidative and energy metabolism and improving the condition of a burn wound without toxic effects on the stomach wall, can be used as an element of additional therapy in the treatment of burns.
Биологическая роль различных ферментов митохондрий может в значительной степени определяться их способностью к связыванию с мембраной, приводя к существенным изменениям структурных и регуляторных особенностей энзимов клетки. Целью данной работы явилось изучение каталитических и кинетических свойств лактатдегидрогеназы (ЛДГ), алкогольдегидрогеназы (АДГ) и комплекса АДГ-ЛДГ митохондрий печени крыс, а также исследование сил электростатического взаимодействия ферментативного комплекса с мембраной митохондрий. В работе были использованы крысы самцы линии Wistar. Активность и кинетические свойства АДГ и ЛДГ определяли в митохондриальной фракции печени крыс. Митохондрии получали методом дифференциального центрифугирования. Для оценки взаимодействия ферментов с мембраной проводилась солюбилизация мембраносвязанных форм ЛДГ, АДГ путем суспендирования митохондрий в 0,15 М NaCl. Показано, что преобладающее количество общей суммарной активности ЛДГ, АДГпр, кластера ЛДГпр-АДГпр интактных животных приходится на лабильно связанную с мембраной форму фермента (солюбилизат). Для АДГобр, надмолекулярного комплекса ЛДГобр-АДГобр крыс выявлено преобладание прочно связанных с мембраной форм ферментов. До солюбилизации и в солюбилизате в составе кластера отмечено снижение удельной активности лактатдегидрогеназы как в прямой, так и в обратной реакциях, увеличение активности АДГпр и АДГобр по сравнению с односубстратными реакциями. Для односубстратных реакций ферментов и энзимов гетерогенной системы (АДГ-ЛДГ) выявлены изменения кинетических характеристик ЛДГ и АДГ, которые приводят к смещению направленности метаболизма митохондрий.
