UVA-induced metabolic changes in non-malignant skin cells and the potential role of pyruvate as antioxidant.

The exposure to UVA (320-400 nm) irradiation is a major threat to human skin concerning photoaging and carcinogenesis. It has been shown that UVA irradiation can induce reactive oxygen species (ROS) and DNA mutations, such as 8-hydroxydeoxyguanosine. Furthermore, UVA induces the expression of photoaging-associated matrix metalloproteases (MMPs), especially of matrix metalloprotease 1 (MMP 1) and matrix metalloprotease 3 (MMP 3). In addition to this, it was recently shown that UVA-induced ROS also increase glucose metabolism of melanoma cells, however, the influence of UVA on the glucose metabolism of non-malignant cells of the human skin has, so far, not been investigated in detail. Here, we investigated the UVA-induced changes in glucose metabolism and the functional relevance of these changes in primary fibroblasts-normal non-malignant cells of the skin. These cells showed an UVA-induced enhanced glucose consumption and lactate production and changes in pyruvate production. As it has been proposed that pyruvate could have antioxidant properties we tested the functional relevance of pyruvate as protective agent against UVA-induced ROS. Our initial experiments support earlier publications, demonstrating that pyruvate treated with H2O2 is non-enzymatically transformed to acetate. Furthermore, we show that this decarboxylation of pyruvate to acetate also occurs upon UVA irradiation. In addition to this, we could show that in fibroblasts pyruvate has antioxidant properties as enhanced levels of pyruvate protect cells from UVA-induced ROS and partially from a DNA mutation by the modified base 8-hydroxydeoxyguanosine. Furthermore, we describe for the first time, that the interaction of UVA with pyruvate is relevant for the regulation of photoaging-associated MMP 1 and MMP 3 expression.

Investigation of the HelioVital filter foil revealed protective effects against UVA1 irradiation-induced DNA damage and against UVA1-induced expression of matrixmetalloproteinases (MMP) MMP1, MMP2, MMP3 and MMP15.

The damaging effects of solar ultraviolet (UV) radiation exposure to human skin are well known and can reach from accelerated skin aging (photoaging) to skin cancer. Much of the damaging effects of solar UVA (320-400 nm) radiation is associated with the induction of reactive oxygen species (ROS), which are capable to cause oxidative damage to DNA like the oxidized guanosine 8-hydroxy-2' -deoxyguanosine (8-OHdG). Therefore, new UV protective strategies, have to be tested for their efficiency to shield against UV induced damage. We investigated the protective effects of HelioVital sun protection filter foil against UVA1 irradiation in skin cells. It could be shown, that HelioVital sun protection filter foil has protective effects against UVA1 irradiation induced changes in matrix metalloproteinase (MMP) expression. Furthermore a UVA1-dependant regulation of MMP15 in human fibroblasts could be shown for the first time in this context. In addition, this study demonstrated the protective effect of the HelioVital filter film against UVA1-induced ROS production and DNA damage. These results could pave the way for clinical studies with HelioVital filter foil shielding against the damaging effects of phototherapy and other forms of irradiation therapy, thereby increasing the safety and treatment opportunities of these forms of therapy.

Association of polymorphous light eruption with NOD-2 and TLR-5 gene polymorphisms.

BACKGROUND: Polymorphous light eruption (PLE) is a common, immunologically mediated, photosensitive skin disease. After ultraviolet-B (UV-B) irradiation, patients with PLE show reduced Langerhans cell (LC) depletion in the epidermis, which results in a non-suppressive microenvironment in the skin. Interestingly, severe acute graft-versus-host disease (aGvHD) occurred in stem cell transplanted patients that showed no or incomplete depletion of LCs after UVB irradiation. Genetic variation in nucleotide-binding oligomerization domain 2 (NOD-2) and toll-like receptor 5 (TLR-5) genes also confers susceptibility to aGvHD. OBJECTIVES: We hypothesized that PLE is associated with genetic variation in the NOD-2 and TLR-5 genes. METHODS: We investigated single-nucleotid polymorphisms (SNPs) of NOD-2 (R702W, G908R, 3020Cins) and TLR-5 (A592S, P616L, N392STOP) in skin biopsies of patients with PLE (n = 143) and in healthy controls (n = 104) using restriction fragment length polymorphism analysis. RESULTS: The frequency of NOD-2 alleles with the SNP R702W was significantly higher in PLE than in controls (31.8% vs. 6.3%; P < 0.0001), and homozygous carriers of this mutation were more common in PLE (27.9% vs. 0%; P < 0.0001). For SNP 3020Cins, the allele frequency (7.3% vs. 0.7%; P = 0.0025) and the number of heterozygotes (14.7% vs. 1.3%; P = 0.0019) were higher in PLE. The frequency of alleles with the N392STOP SNP of the TLR5 gene, which is associated with a truncated, non-functional receptor, was significantly higher in PLE (21% vs. 5%; 7% vs. 1% homozygotes, 28% vs. 8% heterozygotes; P < 0.0001). The other SNPs did not differ significantly. CONCLUSIONS: This study yielded a high frequency of functional SNPs in the NOD-2 and TLR-5 genes in PLE. The same SNPs are associated with aGvHD and there are similarities in the reaction of LCs after UVB irradiation between aGvHD and PLE. This leads to the hypothesis that patients with PLE may be more susceptible to developing GvHD after stem cell transplantation, an assumption that needs to be investigated further.

[Application of modern non-drug technologies to improve the quality of life of patients with hemiparesis]. / Primenenie sovremennykh nemedikamentoznykh tekhnologii dlya povysheniya kachestva zhizni bol'nykh s gemiparezom.

In recent years, the improvement of the system of medical rehabilitation of patients with acute cerebrovascular accident has acquired special social significance due to the high morbidity. OBJECTIVE: To study the effect of the combined use of a multifunctional platform and massage with a pulsed low-frequency electrostatic field on the motor functions of the lower limb, coordination and balance, as well as on the psychoemotional status and quality of life of patients after an ischemic stroke with hemiparesis in the late recovery period. MATERIAL AND METHODS: The article presents data on the treatment of 120 patients who had an ischemic stroke with movement disorders in the form of hemiparesis with an increase in muscle tone by the type of spasticity in the late recovery period. All patients received standard drug therapy and underwent medical rehabilitation. The patients were divided into 3 groups: the main group - 40 patients who received massage with a pulsed low-frequency electrostatic field and training on a multifunctional platform with biofeedback (BFB) - COBS (mtd-Systems, Germany), a comparison group - 40 patients who received training on a multifunctional platform with biofeedback (BFB), control group - 40 patients. RESULTS: In the initial state, all patients included in the study showed an uneven distribution of the load in the protocols «normal standing position¼ and «standing balance¼ with an advantage towards the healthy leg. After the course of treatment in the patients of the main group, all the evaluated indices were normalized, in the patients of the comparison group statistically significant less pronounced results were obtained, and in the patients of the control group only a positive trend was noted. When studying the quality of life and the effectiveness of treatment according to the EQ-5D questionnaire in the examined patients in the initial state, the indicator averaged 7.8±0.8 points. After the course of treatment, the patients of the main group showed a statistically significant decrease in the indicators of the EQ-5D questionnaire from 7.8±0.8 points in the initial state to 5.1±1.1 (p<0.001), which lasted up to 6 months. In patients of the comparison group in all periods of observation, a decrease in the indicator was also noted, but less pronounced, in patients in the control group, only a positive trend. The data were supported by the Hospital Anxiety and Depression Scale (HADS) score. CONCLUSION: The use of complex rehabilitation with the inclusion of a multifunctional platform with biofeedback and electrostatic massage contributes to a pronounced improvement in the functional state of muscle tone, coordination and balance, a significant decrease in the level of anxiety and depression and has a pronounced positive effect on the quality of life and assessment of the health status of patients with hemiparesis in late recovery period after ischemic stroke.

[The use of transdermal electroneurostimulation in the rehabilitation of patients after surgical decompression of the carpal tunnel]. / Primenenie transdermal'noi elektroneirostimulyatsii pri reabilitatsii patsientov posle khirurgicheskoi dekompressii karpal'nogo kanala.

Most of the clinical and neurophysiological disorders of the median nerve that persisted in patients after surgical decompression of the carpal tunnel (DCT) are associated not with acute compression of the nerve, but with myelinoaxonopathic disorders in the structure of the nerve that developed as a result of prolonged compression. OBJECTIVE: To study the effectiveness of transdermal electroneurostimulation (TENS) in the rehabilitation of patients with residual neurological deficit after surgical DCT. MATERIAL AND METHODS: We observed 45 patients after DCT (6-12 months), aged 30 to 50 years (mean age 42±3 years). All patients were diagnosed with primary carpal tunnel syndrome with a lesion of the dominant hand, of which 15 patients received pharmacotherapy; 15 patients - in addition to pharmacotherapy, received a course of low-frequency high-amplitude TENS (NTENS) and 15 patients - a course of high-frequency low-amplitude TENS (VTENS). RESULTS: The regression of positive sensory symptoms was more pronounced against the background of VTENS and NTENS than after the use of pharmacotherapy, on average 6.6 times (p<0.01). At the same time, VTENS turned out to be more effective than NTENS by 28.5% (p<0.05). In the long-term follow-up period (2 months), the severity of positive sensory symptoms decreased in patients who received VTENS and NTENS, by an average of 58.1% compared with the baseline values before treatment (p<0.05). The severity of Tinel's symptom in patients who received a course of VTENS decreased by 44%, after a course of NTENS - by 67%, and after pharmacotherapy - by 14%. It was found that NTENS compared with VTENS was 51% more effective in reducing Tinel's symptom (p<0.05) and by 64% - Falen's symptom (p<0.05). The decrease in the time to perform the Jebsen-Taylor test (JTT) after the use of pharmacotherapy was on average 9% (p>1.00), and after TENS - 23% (p<0.05). When comparing VTENS and NTENS among themselves, NTENS turned out to be more effective on average by 68% (p<0.05). An improvement in electro-kymographic parameters was revealed only after the use of VTENS. At the same time, the decrease in the residual latency was 21.3% and the increase in the amplitude of the M-response was 14.3%. CONCLUSION: Transdermal electroneurostimulation is a highly effective method used in the complex treatment of patients with residual neurological disorders after undergoing surgical decompression of the carpal tunnel. The maximum regression of positive sensory symptoms develops against the background of high-frequency low-amplitude transcutaneous electroneurostimulation, and the maximum anti-inflammatory effect, the greatest improvement in the function of small hand muscles and statistically significant neurophysiological restoration of the function of the median nerve are revealed after the application of a course of low-frequency high-amplitude transcutaneous electroneurostimulation. At the same time, positive dynamics after low-frequency high-amplitude transcutaneous electroneurostimulation and high-frequency low-amplitude transcutaneous electroneurostimulation persists in the long-term period.

[Medical rehabilitation of children with bronchial asthma]. / Meditsinskaya reabilitatsiya detei s bronkhial'noi astmoi.

Medical rehabilitation of children suffering from bronchial asthma is based on the complex use of natural therapeutic factors, drug, non-drug therapy and other methods in order to achieve control over the disease and long-term remission. The use of non-drug technologies is necessary taking into account the pathogenesis of the bronchial asthma development and is aimed at relieving bronchospasm, reducing the activity of allergic inflammation in the bronchi, improving their drainage function, strengthening the respiratory muscles and increasing exercise tolerance. When carrying out rehabilitation measures it is important to observe the basic principles of medical rehabilitation. An individual medical rehabilitation program is drawn up taking into account the assessment of the child's clinical condition at each stage with the diagnosis of the initial rehabilitation status, the establishment of a rehabilitation diagnosis and an assessment of the rehabilitation potential. The article presents the data of the analysis of scientific publications and our own research allowing to draw up an algorithm for the formation of an individual program of medical rehabilitation for children with bronchial asthma.

Expression of coagulin A with low cytotoxic activity by Pediococcus pentosaceus ST65ACC isolated from raw milk cheese.

AIMS: We aimed to evaluate some specific conditions for growth of Pediococcus pentosaceus ST65ACC and its bacteriocin expression through ABC transporters; to purify the bacteriocin and determine its sequence; and to evaluate the cytotoxicity potential of the purified bacteriocin(s). METHODS AND RESULTS: The results presented for growth behaviour of P. pentosaceus ST65ACC showed that the bacterial growth was slightly influenced when cultured in MRS broth with different amounts of inoculum: 1, 2, 5 and 10%. The bacteriocin activity increased when 5 and 10% inocula were used. The carbon source (glucose) used in different amounts (1, 2, 3 or 4%) had no significant effect on growth and bacteriocin production. The studied strain P. pentosaceus ST65ACC was able to metabolize xylooligosaccharide (XOS) as the sole carbon source, resulting in the production of an antimicrobial peptide. The genes involved in the ABC transport system and sugar metabolism of P. pentosaceus ST65ACC were expressed at different levels. The bacteriocin produced by P. pentosaceus ST65ACC was partially purified by precipitation with ammonium sulphate (40% saturation), followed by reversed-phase liquid chromatography, resulting in the identification of an active bacteriocin. Tandem mass spectrometry was used to identify the partial sequence KYYGNGVTCGKHSCSVDWGK sharing high similarity to coagulin A. The semi-purified bacteriocin had low cytotoxicity based on estimated values for maximal nontoxic concentration (MNC) and cytotoxicity concentration (CC50 ). CONCLUSIONS: The bacteriocin produced by P. pentosaceus ST65ACC is similar to coagulin, with low cytotoxicity, strong antimicrobial activity and possible additional metabolite routes in the producer cell. In addition to MRS broth, bacteriocin was produced also in medium containing XOS (as the single carbon source). SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report of evaluation of the role of ABC transporters in the expression of bacteriocin by P. pentosaceus, cultured in MRS and XOS.

Immunohistochemical and morphological characteristics of tissues response to polylactic acid membranes with silver nanoparticles.

BACKGROUND: The aim of this research was to study anti-microbial and anti-inflammatory characteristics of silver nanoparticles helping bone structures to recover during late stage of parodontitis, which afterwards will increase the effect of bone regeneration operations. MATERIALS AND METHODS: We assessed colloid solution-derived silver nanoparticles coating of polylactic acid membrane regarding tissue foreign body response. Thirty eight polylactic acid membranes were implanted intracranially in rabbits - ten unmodified (control group) and twenty eight with silver nanoparticles coating (experimental group). In controls, penicillin was used for infection prophylaxis. Tissue response was assessed by light microscopy and immunohistochemistry (CD3, CD15, CD30) 2 weeks after implantation. RESULTS: inflammation markers in experimental group were significantly lower than in control group, there were no signs of forming a fibrosis capsule nor infectious signs. CONCLUSIONS: colloid silver solution can be used as a source of nanoparticles for anti-microbial and anti-inflammatory biodegradable membranes' coating.

[Myostimulating effect of comprehensive application of general magnetotherapy, fractional CO2 laser, electro-myostimulation and special complex of exercise in patients with rectoceleal after open-peconstructure]. / Miostimuliruyushchii effekt kompleksnogo primeneniya obshchei magnitoterapii, fraktsionnogo CO2-lazera, elektromiostimulyatsii i spetsial'nogo kompleksa LFK u patsientok s rektotsele posle rekonstruktivno-plasticheskikh operatsii.

The article presents data on the treatment of 200 women of childbearing, peri - and menopausal age with rectocele of II-III degree, which were divided into 4 groups comparable in clinical and functional characteristics: 1-50 patients who, after surgical treatment of rectocele, underwent a complex consisting of a course of General magnetic therapy, 2 intra-vascular procedures of fractional microablative CO2 laser therapy, electromyostimulation with the pelvic floor muscles and a special complex of therapeutic physical education; comparison 1, which included 50 patients who underwent the above-mentioned complex of rehabilitation measures without General magnetotherapy; comparison of 2-50 patients after surgical treatment of rectocele, who in the late postoperative period (one month after the operation) underwent a set of rehabilitation measures, including a course of electromyostimulation with the biological connection of the pelvic floor muscles, consisting of 10 daily procedures and a special complex of physical therapy and a control group - 50 patients after surgical treatment of rectocele, who in the late postoperative period were treated with symptomatic therapy, including painkillers and antispasmodics, which served as a background for all other groups. As a result of the research, it was found that the developed rehabilitation complexes have a pronounced myostimulating effect, and can be recommended for wide use in rehabilitation programs for postoperative management of patients with rectocele.

[To the question of the possibility of using selective chromotherapy for allergic diseases in children]. / K voprosu o vozmozhnosti primeneniya selektivnoi khromoterapii pri allergicheskikh zabolevaniyakh u detei.

Allergic diseases are a common pathology in childhood. In the comprehensive medical rehabilitation of children with allergic pathology, non-drug methods of treatment are widely used, which help to reduce the number of drugs used, achieve and prolong the remission of the disease, favorably affect the clinical and functional indicators. THE PURPOSE OF THE STUDY: Is the scientific justification for the use of selective chromotherapy in children with bronchial asthma (BA) and atopic dermatitis (AD). MATERIAL AND METHODS: The study included 120 children with allergic diseases (BA and AD). Among 100 patients with BA, the main group included 50 children, who received exposure to monochromatic polarized green light on biologically active zones for 10 days, 50 - a comparison group that did not receive physiotherapy. The main group of children with AD included 10 patients who received selective blue chromotherapy for foci of skin lesions; the comparison group included 10 children who used only moisturizers without physiotherapy methods. In order to assess the effectiveness of the impact of physical factors in all patients, clinical and functional studies were conducted in the dynamics before and after treatment. RESULTS AND DISCUSSION: The results of clinical and functional examinations showed high therapeutic efficacy of the use of monochromatic polarized green light in children with BA (92.0%). The effectiveness of treating children with blood pressure AD using monochromatic polarized light (blue) was 80%. Indications for the use of selective chromotherapy in children with allergic diseases have been developed. For children with BA, selective chromotherapy of the green spectrum is indicated in the presence of a mild to moderate course of the disease, a period of incomplete remission, and an increased level of anxiety. It is advisable to prescribe selective chromotherapy of the blue spectrum to children with a moderate and mild course of AD. CONCLUSION: The positive effect of selective chromotherapy of the green spectrum on the clinical course of BA in children, bronchial patency, the functional state of the central nervous system and its autonomic part, and the psychoemotional status of children have been identified. The use of selective blue spectrum chromotherapy in children with AD helps to decrease the severity of objective symptoms, reduce the intensity of itching and sleep disturbance, as well as a marked decrease in the area of skin lesion.

[Cardiomyoblasts Produced from Mesenchymal Stem Cells in Complex Therapy of Heart Radiation Damage].

This article describes the results of systemic transplantations of cardiomyoblasts grown from autologous or allogeneic bone marrow-derived mesenchymal stem cells, in the complex therapy of the late radiation da- mage of the heart, which developed after radiation therapy in 16 female patients with Hodgkin disease (HD) or breast cancer (BC). The cell therapy drastically improved the efficacy of the drug treatment, which earlier was the only option for the therapy of the radiation damage of vital organs. The effect of such therapy was clinically observed in the patients already in the first year of observation, and consisted in the decrease of the degree of the cardiac failure severity and improvement of their quality of life in the absence of HD or BC progression.

Proteolytic activity of Enterococcus faecalis VB63F for reduction of allergenicity of bovine milk proteins.

With the aim of screening proteolytic strains of lactic acid bacteria to evaluate their potential for the reduction of allergenicity of the major bovine milk proteins, we isolated a new proteolytic strain of Enterococcus faecalis (Ent. faecalis VB63F) from raw bovine milk. The proteases produced by this strain had strong activity against caseins (αS1-, αS2-, and ß-casein), in both skim milk and sodium caseinate. However, only partial hydrolysis of whey proteins was observed. Proteolysis of Na-caseinate and whey proteins, observed after sodium dodecyl sulfate-PAGE, was confirmed by analysis of peptide profiles by reversed-phase HPLC. Inhibition of proteolysis with EDTA indicated that the proteases produced by Ent. faecalis VB63F belonged to the group of metalloproteases. The optimal conditions for their activity were 42°C and pH 6.5. The majority of assessed virulence genes were absent in Ent. faecalis VB63F. The obtained results suggest that Ent. faecalis VB63F could be efficient in reducing the immunoreactivity of bovine milk proteins.

[Contemporary potential of nonmedical treatment in ophthalmology].

The article covers treatment results of 90 patients with primary glaucoma simplex of stage 1 and 2 (137 eyes). The authors proved that using a specified magnetotherapy method, when compared to regular medical therapy, causes more fast and marked regression in clinical symptoms and improvement of vision in patients with primary glaucoma simplex of stages 1 and 2.

[THE APPLICATION OF DOT-TECHNIQUE FOR DETECTING ANTIGENS OF ADENOVIRUS IN CLINICAL SAMPLES].

The article substantiates possibility of application of point enzyme-linked immunosorbent assay (dot-technique) for detecting viral antigens in samples from patients. To diagnose adenovirus infection conjugate of virus-specific monoclonal antibodies and peroxidase of horse-radish were used The chromatographic rectification of conjugate from free peroxidase permits diminishing background coloring of nitrocellulose membrane and therefore to increase sensitivity. The application of direct conjugates on the basis of virus-specific monoclonal antibodies increases specifcity of dot-technique and significantly shortens time period of analysis. As in case of application of direct conjugates on the basis of polyclonal serum, samples from patients require preliminary processing with detergent for preventing non-specific reactions. The dot-technique demonstrates good coincidence with data of polymerase chain reaction and after clinical trials it can be used in diagnostic of human viral infections.

[Approaches to phototherapy in children with chronic dermatosis (review of literature)].

The article covers main aspects of phototherapy and photochemotherapy in treatment of skin diseases in children, risks of such methods usage, prophylactic and therapeutic measures, and precaution means for such treatment in pediatrics.

[Investigation of Free Radical Processes in Substrates and Biological Samples by Means of induced Chemiluminescence].

The possibility of applying induced chemiluminescence was investigated to evaluate oxidative capability of the substrate under the action of hydroxyl radicals and estimate the intensity of free radical processes in biological samples based on the analysis of the organic hydroperoxyde response in a probe. For this purpose the chemiluminescence light sum was measured in 3 steps: when Fe2+ was introduced into in a sample; in course of Fenton reaction (introducing Fe2+ and H202) and when Fe2+ was introduced into the sample after Fenton reaction. Light sum was measured depending on concentration (dilution) of the sample. It was shown that the light sum reaches the maximum value at certain dilution of the substrate studied. The maximum chemiluminescence's position is determined by concentration of RH fragments being oxidized, but the chemoluminescence light sum value is determined partially by the inhibitor [InH]/[RH] and organic hydroperoxides [ROOH]/[RH] found in the sample.

[SOME ASPECTS OF NON-SPECIFIC PROPHYLAXIS AND THERAPY OF ESPECIALLY DANGEROUS INFECTIONS].

Recently, due to spread of dangerous and especially dangerous infections much attention is given to development of complex approaches to their prophylaxis and therapy. Data on use of immune modulators, cytokines, probiotics, preparations of plant origin for non-specific prophylaxis of especially dangerous infections are analyzed in the review, and expediency of their combined use with specific and emergency prophlaxis of these diseases is evaluated.

[EPIZOOTOLOGICAL CHARACTERIZATION OF TREMATODIASIS FOCI IN THE ECOSYSTEM OF THE AMUR RIVER IN THE KHABAROVSK TERRITORY].

Investigations were conducted in late June to early June 2013. The population density of mollusks of the genus Juga in the shoreline and littoral covered by meadow waters was 10 to 30 specimens/m2; there were solitary specimens of the genus Parafossarulus. Fourteen out of 39 fish species were invaded by metacercariae of Clonorchis. Manchurian spiny loaches (Leptobotia) and Light's bitterling (Rhodeus lighti) are first registered to be hosts of Clonorchis. The metacercariae of Metagonimus yokogawai were detected in silver carp (Hypophthalmichthys molitrix) and those of Nanophyetus salmincola schikhobalowi were in the kidneys, fin muscles, and gills of taimen (Hucho) and lenok (Brachymystax lenok) from the Khor River. The invasion rate for the taimen was greater than 1000 metacercariae and that for the lenok was not more than 720 metacercariae per fish.