[Possibilities of using recombinant human deoxyribonuclease I in otorhinolaryngology]. / Vozmozhnosti primeneniya rekombinantnoi dezoksiribonukleazy I cheloveka v otorinolaringologii.

ANNOTATION: Dornase alfa (Pulmozyme, Tigerase) is a purified solution of recombinant human DNase, clinically developed for the treatment of pulmonary diseases in patients with cystic fibrosis (CF). The action of the drug is aimed at destroying the viscous secretion, rich in DNA strands of neutrophils, through their fragmentation, the density of the secretion decreases, and the aeration of the lower respiratory tract improves. The similarity of pathological processes with the formation of viscous exudate on the surface of the mucous membrane in diseases of the upper respiratory tract and ear initiated studies on the use of Dornase alpha in otorhinolaryngology. MATERIAL AND METHODS: The analysis of materials of domestic and foreign authors on the effectiveness of the use of the drug Dornase alfa in otorhinolaryngology was carried out. RESULTS: The review included 132 patients (10 studies) in whom Dornase alfa was used to treat CF-associated nasal and paranasal sinus diseases. Analysis of the literature revealed only 3 studies, one of which consisted of two parts, examining the effect of Dornase alpha on middle ear exudate: two studies were demonstrated in an animal model; one - in vitro on samples of middle ear effusion which were aspirated through a myringotomy incision from patients with recurrent acute otitis media; and one in clinical 40 patients (40 ears) for hydrolysis of exudate in the tympanostomy tubes. CONCLUSION: Analysis of studies on the use of Dornase alfa demonstrates an improvement in clinical symptoms in all patients with CF and chronic rhinosinusitis. In experimental studies on an animal model, as well as in vitro research on exudate from the middle ear, Dornase alfa has demonstrated high efficacy and safety. Dornase alfa is a drug with high potential, further research is needed for wider use in ENT practice, especially in otiatrics.

Transcriptional Activity of Some Genes Involved in Apoptosis in Patients with Vulvar Lichen Sclerosus.

Lichen sclerosus of the vulva is a common, but poorly studied disease. We assessed the level of transcriptional activity of APAF1, BAX, BCL2, BIRC5, CCND1, DAPK1, MCL1, and MYC genes encoding products that control apoptosis in the samples of tissues affected by vulvar lichen sclerosus and adjacent control tissues (n=24). Analysis of transcriptional activity was performed by real-time PCR using specific primers and SYBR Green intercalating dye. After the total group was divided by the presence of the concomitant gynecological diseases, a significant increase in the transcriptional activity of the CCND1 gene was revealed in patients with concomitant uterine fibroids. This may indicate the possible role of the activation of mitosis during tumor initiation.

IL-6 Gene Expression as a Marker of Pathological State in Psoriasis and Psoriatic Arthritis.

The expression of the IL-6 gene in mononuclear blood cells of 45 patients with psoriatic arthritis and 31 patients with plaque psoriasis was studied for possible differential diagnosis of the pathologies. The expression level of IL-6 in psoriatic arthritis and psoriasis surpassed that in healthy controls by 192 and 147 times, respectively. Significant differences in the gene expression were revealed between the patients with psoriatic arthritis and mild psoriasis. The level of IL-6 in patients with severe psoriasis approached that in patients with psoriatic arthritis. High level of IL-6 gene expression can be a marker of possible joint damage in patients with psoriasis and a signal for revising the therapeutic approach in a particular patient.

[Morphochemical study of alpha-synuclein, iron and iron-containing proteins in the substantia nigra of the brain in Parkinson's disease]. / Morfokhimicheskoe issledovanie al'fa-sinukleina, zheleza i zhelezosoderzhashchikh belkov v chernom veshchestve golovnogo mozga pri bolezni Parkinsona.

OBJECTIVE: To study, using a complex morphochemical approach, the localization of alpha-synuclein, iron compounds and iron-containing proteins in the structures of the substantia nigra of the brain in Parkinson's disease (PD). MATERIAL AND METHODS: Histochemistry and immunohistochemistry methods have been used to study the localization of pathological alpha-synuclein (α-Syn-p129), iron compounds and iron-containing proteins - transferrin receptor and ferritin in neurons and neuroglia in the substantia nigra of the brain of deceased PD patients and persons with no neurological symptoms detected during life (control). RESULTS: In the substantia nigra of PD patients, in comparison with the control, a stable accumulation of pathological alpha-synuclein (α-Syn-p129) in the bodies and processes of neurons was found, and in the neuroglia and neuropil - the accumulation of iron (II) and ferritin heavy chain, the reaction of microglia to protein CD68 was moderately elevated. The transmembrane protein CD71 was detected equally in the brains of PD patients and in controls. CONCLUSION: Synaptic protein alpha-synuclein in PD turns into a pathological metabolite that accumulates in the structures of substantia nigra, and probably disrupts the conduction of nervous excitation. Excessive accumulation of the ferritin heavy chain in neuroglia can increase the concentration of reactive forms of iron and increase neurotoxicity. The uniform distribution of the transmembrane glycoprotein CD71 in the of substantia nigra structures both in the control and in PD patients indicates the preservation of non-heme iron transport during the neurodegenerative process.

Genetic Markers of Therapeutic Efficacy of Methotrexate in Patients with Psoriasis.

We studied the effect of C677T and A1298C polymorphisms of the MTHFR gene and 2R/3R polymorphisms of the TYMS gene on the sensitivity to methotrexate in patients with psoriasis (n=139). It was shown that genotype 3R/3R TYMS (OR 8.86, 95%CI 2.00-39.22) and complex genotypes MTHFR1298:A;TYMS:3R (OR 8.20, 95%CI 2.36-28.48) and MTHFR677:C;TYMS:3R (OR 5.40, 95%CI 1.95-14.94) were associated with sensitivity to methotrexate, while genotype 2R/2R TYMS (OR 8.20, 95%CI 2.36-28.48) and complex genotypes MTHFR1298:C;MTHFR677:T;TYMS:2R (OR 0.18, 95%CI 0.06-0.56) and MTHFR1298:C;MTHFR677:T (OR 0.23, 95%CI 0.09-0.59) were associated with resistance to methotrexate. The results can be used for preventive assessment of the effectiveness of methotrexate treatment in patients with psoriasis.

LC-MS/MS analysis of lesional and normally looking psoriatic skin reveals significant changes in protein metabolism and RNA processing.

BACKGROUND: Plaque psoriasis is a chronic autoimmune disorder characterized by the development of red scaly plaques. To date psoriasis lesional skin transcriptome has been extensively studied, whereas only few proteomic studies of psoriatic skin are available. AIM: The aim of this study was to compare protein expression patterns of lesional and normally looking skin of psoriasis patients with skin of the healthy volunteers, reveal differentially expressed proteins and identify changes in cell metabolism caused by the disease. METHODS: Skin samples of normally looking and lesional skin donated by psoriasis patients (n = 5) and samples of healthy skin donated by volunteers (n = 5) were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After protein identification and data processing, the set of differentially expressed proteins was subjected to protein ontology analysis to characterize changes in biological processes, cell components and molecular functions in the patients' skin compared to skin of the healthy volunteers. The expression of selected differentially expressed proteins was validated by ELISA and immunohistochemistry. RESULTS: The performed analysis identified 405 and 59 differentially expressed proteins in lesional and normally looking psoriatic skin compared to healthy control. In normally looking skin of the patients, we discovered decreased expression of KNG1, APOE, HRG, THBS1 and PLG. Presumably, these changes were needed to protect the epidermis from spontaneous activation of kallikrein-kinin system and delay the following development of inflammatory response. In lesional skin, we identified several large groups of proteins with coordinated expression. Mainly, these proteins were involved in different aspects of protein and RNA metabolism, namely ATP synthesis and consumption; intracellular trafficking of membrane-bound vesicles, pre-RNA processing, translation, chaperoning and degradation in proteasomes/immunoproteasomes. CONCLUSION: Our findings explain the molecular basis of metabolic changes caused by disease in skin lesions, such as faster cell turnover and higher metabolic rate. They also indicate on downregulation of kallikrein-kinin system in normally looking skin of the patients that would be needed to delay exacerbation of the disease. Data are available via ProteomeXchange with identifier PXD021673.

Association of the NOS3 Gene Polymorphisms with the Risk of Atopic Dermatitis in Children.

We studied association of polymorphic markers Glu298Asp (rs1799983), C774T (rs1549758), and T786C (rs2070744) of the NOS3 gene with the risk of atopic dermatitis. It was found that T786C polymorphic marker of the NOS3 gene is associated with lower risk of erythematosquamous lichenoid atopic dermatitis. A significant association between the homozygous CC genotype in locus 786 of the NOS3 gene and the development of erythematosquamous atopic dermatitis with lichenification was revealed. The homozygous CC genotype can be considered as a risk factor of erythematosquamous atopic dermatitis with lichenification.

SkQR1 Reduces Neurologic Deficit Caused by Rat Brain Compression Ischemia.

The protective effect of antioxidant SkQR1 was examined on the model of left-sided compression ischemia in rat sensorimotor cortex. The special tests aimed to determine the neurologic deficit in the limbs and assess performance of the forelimbs showed that a 2.5-min ischemia produced no disturbance in the limb functions on postsurgery days 1, 3, and 7. Elevation of compression time resulted in neurologic deficit in animals, and its severity depended on this time. A single intravenous injection of SkQR1 (250 nmol/kg body weight) performed 30 min after ischemia significantly reduced the degree of neurologic deficit. In vitro model of ischemia in surviving rat hippocampal slices showed that a 15-min-long ischemia significantly inhibited the population excitatory postsynaptic potentials, which did not restore during reperfusion. Preincubation of the slices with SkQR1 did not significantly affect recovery of these potentials.

Interactions between nanoparticles in nanosuspension.

Nanoparticles are particles with a characteristic dimension below 100 nm. The properties of nanoparticles differ substantially from those of "big" colloidal particles (size bigger than 1 µm) because radius of surface forces, which is around 100 nm, is greater than or comparable with the nanoparticles size. The latter means that each nanoparticle could be completely covered by the surface forces of the neighbouring particles at small enough separation. It also means that the well-known Derjaguin approximation cannot be applied directly and some modifications are required. Pairwise interaction between nanoparticles can be used only at an extremely low volume fraction of nanoparticles (below some critical volume fraction, which is ~0.02%), and above this concentration a new theory based on many-particle interactions should be applied, which is yet to be developed. Some recent progress in the area of interaction between nanoparticles is reviewed and the properties of nanosuspensions based on interaction between nanoparticles are described. The authors have not attempted to cover all available literature in the area but instead have tried to underline the fundamental problems in the area which need to be addressed.

The role of regulatory RNAs (miRNAs) in asthma.

INTRODUCTION: Recently, a great deal of attention has been paid to the investigation of regulatory functions of microRNA. Currently, many different mechanisms involved in the pathogenesis of asthma are known, but the whole picture of pathogenesis has not yet been studied. CONCLUSIONS: MicroRNAs play an important role in the regulation of many cellular processes. Undoubtedly, these regulatory molecules are involved in the pathogenesis of asthma, and therefore can be potential targets for treatment.

[Morphochemical changes in the substantia nigra cellular structures in Parkinson's disease]. / Morfokhimicheskie izmeneniia kletochnykh struktur chernogo veshchestva golovnogo mozga pri bolezni Parkinsona.

AIM: to clarify the features of morphochemical changes in the substantia nigra cellular structures in Parkinson's disease. MATERIAL AND METHODS: The structural characteristics of the substantia nigra were studied microscopically and quantified using computer morphometric methods at brain autopsies of individuals with Parkinson's disease who had died from intercurrent diseases and those who had no evidence of neurological disorders in their history (a control group). RESULTS: This investigation could clarify the features of morphochemical changes in both the neural network structures and the glial populations of the substantia nigra in Parkinson's disease. The number of neurons containing tyrosine hydroxylase (a marker of dopamine neurons) in the compact part of the substantia nigra (a ventral region) was smaller and the density distribution of Lewy bodies was higher in the patients with Parkinson's disease than in the control group. The accumulation of iron (II) compounds in the cellular elements and neuropile and the increased expression of glial fibrillary acidic protein in Parkinson's disease were more pronounced than those in the controls. CONCLUSION: Postmortem diagnosis in Parkinson's disease should be based on a full description of a set of neuronal and glial morphochemical and structural changes in the substantia nigra rather than on the identification of cellular markers for the neurodegenerative process.

Wetting films of aqueous solutions of Silwet L-77 on a hydrophobic surface.

The formation of wetting films of aqueous solutions of Silwet L-77 on hydrophobic substrates takes place only at concentrations above the critical aggregation concentration (CAC). At concentrations above the critical wetting concentration (CWC) a new phenomenon was found: the formation of multilayered spots of thicker films in the wetting film of aqueous solutions of Silwet L-77 on hydrophobic surfaces. An expansion of the thicker spots within the film and the formation of "channels" between the spots and the edge of the film led to a continuous shrinkage of the wetting film and its disappearance in the end. We suggested that the multiple thicker films originate from the multilayer structuring of trisiloxane bilayers within the wetting film.

[DEPENDENCE OF FUNCTIONAL PARAMETERS OF RAT SKELETAL MUSCLE CONTRACTION FROM LEVEL CIRCULATING TRIIODOTHYRONINE].

In experiments on white rats with different thyroid status (thyroidectomy, euthyroidism, hyper- and a thyrotoxicosis) in conditions in situ the relationship between the severity of functional parameters of the tibialis anterior muscle and individual level of circulating free triiodothyronine was learnt. The latent period of muscle contraction, the average and maximum speed of its contraction were determined. A comparative quantitative analysis of the values of indicators of the contractile function of the tibialis anterior muscle in euthyroid and thyroidectomy rats allowed to evaluate the «thyroid contribution) of free triiodothyronine in the functional state of the native skeletal muscle, which formed to 15% for the latency reduction period, 12 % for the average speed of contractile act and 8% for of the maximum speed of muscle contraction. It is shown that in rats if the control group at the beginning of the scale of triiodothyronine concentration (2.2-4.9 pmol/L) hormone activity is expressed weakly, and in the range of 5.2-7.6 pmol/L is greatly increased (up to 11 times). It was established that the positive effect of triiodothyronine is preserved far beyond of the physiological norm of hormone concentration. It is allowed to speak about the existence of an extended corridor of hormone activity. On the base of the analysis of the degree of severity of the physiological parameters of the muscle contraction curve from circulating level of triiodothyronine in rats with different thyroid status (0.1-45 pmol/L) constructed of functional activity of free triiodothyronine. This curve consists of 7 different zones which are located in different sectors of the scale of concentration.

Influence of iatrogenic hypercorticoidism induced by long-term application of dexamethasone on power of muscular contraction of white rats.

Research objective consisted in detection of nature of the changes of the myothermiс and the ergometric parameters of the contraction of the forward tibial muscle of rats in the course of performing of the tiring work at the saturation of an organism by therapeutic doses of dexamethasone. Method: The experiments were performed on sexually mature rats-females (200-220 g), divided into control (n = 10) and experimental (n = 60) groups. The animals of experimental group received dexamethasone (D, KRKA, Slovenia) in a dose of 0,25 mg/kg (intraperitoneal, 1 time in 2 days) during from 10 to 60 days. On anesthetized animals (sodium thiopental, 100 mg/kg) with the use of myothermia and ergographia the nature of change of power of the muscle's contraction in the course of the performance of the tiring work (3 six-second tetanus with external loading of 80 g) was studied. Restults: At the initial stage of the development of iatrogenic hypercorticoidism (after 5-20 injections of D) the initial value of the external work of the muscle in comparison with the control is significantly lower (for 30-52%) and the temperature cost of the unit of the work (TCMW), on the contrary, - is higher (for 26-82%). On the end of the 2-month period of application of D the initial values of the power parameters of the muscle came back to control level. During the performance of the tiring tetanus in animal experimental groups the decline of the external work of the muscle is greater (69-73%) compared with the control (55%). This effect does not depend of the number of injections of D, which indicates about a high pathophysiological activity of glucocorticoid concerning working capacity of the muscle. At expressed fatigue the TCMW always increases from 104% (5 injections of D) to 230% (20 injections); at control animals the effect of the tiring work on TCMW is significantly weaker (28%). At long-term application of D (2 months) the described effect of the preparation is weakened, though remains accurately expressed. Conclusion: The obtained data are considered from the point of view of formation at the hypercorticoidizm of the pathophysiological mechanism - the increase of power cost of muscular work. The revealed effect of D can be the cornerstone of the formation of the number of the pathophysiological mechanisms in neuromuscular system including causing the development of the myopathy at the hypercorticoidizm.

[AMPLITUDE-FREQUENCY'S DEPENDENCE OF M-RESPONSE OF THE SKELETAL MUSCLE OF RATS WITH EXPERIMENTAL HYPERCORTICOIDIZM].

In experiments over the mature white female rats the influence of the hypercorticoidizm (simulated by daily parenteral injection of hydrocortisone in a dose of 3 mg/kg/days for 30 days) on some parameters of the M-response of the forward tibial muscle with a different frequency of stimulation of the low-tibial nerve is studied. It is established that the hypercorticoidizm is followed by lengthening of the chronaxia of the forward tibial muscle at its indirect irritation (by 69 per cent), deterioration of stability of M-response's generation, lengthening of the latent period (by 30 per cent) and to reduction of amplitude (by 29 per cent) of single M-responses against increase in frequency of polyphase potentials (to 35 per cent). At animals with hypercorticoidizm in the range of low frequencies of nerve's stimulation (10-30 imp/s) periodic generation of higher-amplitude M-responses, than at control, against their low initial amplitude was observed, which can testify in a favor of an initial partial blocking of synapses. The hypercorticoidizm was followed more expressed, in comparison with control, decreasing of M-responses' amplitude in the process of increasing in frequency of low-tibial nerve's stimulation, decreasing in frequency of nerve's stimulation on achievement which inverse relationship between M-responses' amplitude and frequency of nerve's irritation was established.

[THE QUANTIFICATION OF SULPHATED GLYCOSAMINOGLYCANS IN RAT URINE IN EXPERIMENTAL HEMORRHAGIC CYSTITIS].

The paper presents the study of the excretion of sulfated glycosaminoglycans (GAG) in the urine of rats in experimental hemorrhagic cystitis induced by cyclophosphamide and treated with glycosaminoglycan replacement therapy. Rats were given intraperitoneal injections of cyclophosphamide at a dose of 100 mg per 1 kg body weight and subsequently treated with intragastric administration of the combined preparation of glycosaminoglycans containing glucosamine hydrochloride and chondroitin sulfate at a dose of 10 and 100 mg per 1 kg of body weight. Within 24 or 72 hours after cystitis induction there was a statistically significant increase in urinary GAG excretion. The study also found a decrease (from 1.34 to 1.22 mg/dL) in urinary GAG within 0 to 72 hours following induction of acute cystitis without glycosaminoglycan therapy. In the subchronic model of inflammation in the bladder, upon repeated administration of low doses of cyclophosphamide (50 mg/kg), decrease in urinary GAG within 0 to 72 hours (1,32±0,13 mg/dL) as well as increased excretion after 96 hours at a concentration of 2,29±0,13 mg/L after initiation cystitis were found.

[The results of a comparative randomized study of the effectiveness of various methods for the correction of the nasal valve: the objective evaluation].

The present comparative randomized controlled study was designed to evaluate the effectiveness of the most widely used methods for the surgical treatment of nasal valve dysfunction including sutural extension of the valve, stretching the lateral nasal wall in combination with the application of strengthening transplants, and the introduction of tissue expanders. The objective studies were carried out with the use of anterior active rhinomanometry and acoustic rhinometry performed within 1, 3, and 12 months after surgery. It was shown that the correction of dysfunction of the nasal valve with the application of expanding transplants is the most effective method for the normalization of the parameters of nasal breathing during the mid- and long-term follow up. Plastic surgery with the use of local tissues produces a less pronounced but stable beneficial effect whereas the sutural extension of the valve yields only short-term positive results. On the whole, the effectiveness of the latter approach is inferior to that of the former two methods.

[The results of a comparative randomized study on the effectiveness of various methods for the correction of the nasal valve: the subjective evaluation].

The objective of the present study was to compare the effectiveness of several extensively used techniques for the correction of the nasal valve in terms of the main clinical manifestations with the application of various scales for subjective evaluation of the outcomes of the treatment. It was demonstrated in the course of this prospective study that the patients described the elimination of nasal valve dysfunction with the help of expanding transplants as the most effective method in terms of the improvement of nasal breathing and the achievement of the acceptable aesthetic results. The plastic correction with the use of local tissues was reported to be less efficacious even if ensuring the stable result. This method did not worsen the shape of the nose but failed to remove its existing cosmetic defect. As far as the aesthetic outcome of the treatment is concerned, the suture correction technique was recognized to be the least efficacious approach because it resulted in the deterioration of the nose shape in more than half of the cases.

[METHOD FOR QUANTITATIVE EVALUATION OF THE INTENSITY OF THE HISTOCHEMICAL REACTION FOR SULFATED GLYCOSAMINOGLYCANS IN THE TISSUES OF RAT URINARY BLADDER].

In this paper a method is proposed for the quantitative evaluation of the concentration of sulfated glycosaminoglycans in the tissues of rat bladder using the calibration standards of alcian blue (AB) in 20% aqueous solution of gelatin. The blocks that have the consistency of a dense gel and serve as calibration standards of different dye concentrations, sections 7 and 9 µm thick are cut, and photographed. Using the "Videotest 5.0" program, in the slices with different concentrations of the dye, relative optical density of AB was determined and the graph of its dependence on dye concentration was plotted. On the basis of a calibration graph, it was possible to determine AB concentration in the histological sections of the urinary bladder stained with AB. In the sections of the urinary bladder of the intact rats the concentration of AB in the mucus covering the epithelial layer, was 22 ± 1 3 mg/cm3, while in the tissues of its lamina propria it was equal to 2.5 ± 1.0 mg/cm3.

Effects of activated protein C on the size of modeled ischemic focus and morphometric parameters of neurons and neuroglia in its perifocal zone.

The effects of activated protein C (APC) on the quantitative parameters of neurons and neuroglia in the perifocal zone of infarction induced in the left hemispheric cortex were studied in two groups of rats. Group 1 animals served as control (control infarction). Group 2 rats were injected with APC (50 µg/kg) in the right lateral cerebral ventricle 3 h after infarction was induced, and after 72 h the infarction size was evaluated and the neurons and neuroglia in the perifocal zone were counted. APC reduced the infarction size 2.5 times in comparison with the control and reduced by 16% the neuronal death in the perifocal zone layer V, causing no appreciable changes in layer III, and did not change the size of neuronal bodies but increased (by 11%) the size of neuronal nuclei in layer III. The protein maintained the sharply increased count of gliocytes in the perifocal zone of infarction and promoted their growth. Hence, APC protected the neurons from death in the ischemic focus by increasing the gliocyte count and stimulating the compensatory reparative processes.