TGFB1, FOXO1, and COMP Genes Expression in Blood of Patients with Osteoarthritis after SARS-CoV2 Infection.

Abstract-Nowadays the possible influence of the coronavirus infection on cartilage degeneration and synovial membrane inflammation during chronic joint pathology-osteoarthritis-remains largely unelucidated. The aim of the presented work is to analyze the TGFB1, FOXO1, and COMP gene expression and free radical generation intensity in blood of patients suffering from osteoarthritis after beating the SARS-CoV2 infection. The work was carried out using molecular genetics and biochemistry methods. The decrease of the TGFB1 and FOXO1 expression level was shown to be more evident in the osteoarthritis patients after COVID-19 if compared to the group with knee osteoarthritis during simultaneous and more prominent diminishing of both superoxide dismutase and catalase activity (possibly indicating cell redox state disruption and TGF- P1-FOXO1 signaling attenuation) in patients with osteoarthritis after SARS-CoV2 disease. At the same time, the more prominent decrease of COMP gene expression level was demonstrated in patients with osteoarthritis after COVID-19 compared to the group with knee osteoarthritis and more intense increase of the COMP concentration in patients with osteoarthritis after the SARS-CoV2 infection was revealed. These data indicate more significant activation of cell destructive processes after the infection as well as further pathology progression.

WAYS TO IMPROVE THE EFFICACY OF ORTHOPEDIC TREATMENT OF PATIENTS WITH SEVERE EXCESSIVE TOOTH WEAR.

Excessive tooth wear is currently one of the main problems of oral health. Excessive tooth wear affects both function and aesthetics. Teeth with excessive tooth wear cannot be effectively used for biting and chewing food. Treating excessive tooth wear is complex and time consuming. Studies on increasing the effectiveness of treatment of such patients are relevant. The study involved 60 people with a significant degree of excessive tooth wear and a decrease in the height of the bite. We examined patients using measurements of the electromyographic (EMG) activity of mm.masseter and temporalis. The subjects were divided into two groups - in the first group (30 people), the maxillofacial system was prepared for further prosthetics by wearing a removable dental splint-teethguard, in the second group (30 people) we used both removable splint-teethguards and transcutaneous electrical nerve stimulation (TENS). In both groups, an electromyographic study of mm.masseter and temporalis was performed after 1 week, 1 and 2 months of treatment. In patients with excessive tooth wear and a significant decrease in bite height, our method of preparation for orthopedic treatment contributed to a more rapid normalization of mm.masseter and temporalis functions. The results indicate a pronounced positive effect of the use of removable teeth guard and TENS on the normalization of the bioelectric activity of the masticatory muscles in the preparation of patients for orthopedic treatment of excessive tooth wear.

A SORAFENIB INDUCED MODEL OF GLOMERULAR KIDNEY DISEASE.

Glomerular injury and proteinuria are important pathophysiological features of chronic kidney disease. In the present study, we provide data on a glomerular injury model that was developed using the cancer chemotherapy drug sorafenib. Sorafenib is a tyrosine kinase inhibitor that acts via the vascular endothelial growth factor (VEGF) signaling pathway and is widely used to treat a variety of cancers. On the other hand, sorafenib causes serious renal side effects in patients including the development of chronic kidney disease. The current study aimed to utilize the nephrotoxic property of sorafenib to develop a rat model for chronic kidney disease. We demonstrate that rats administered sorafenib for 8 weeks along with a high salt diet (8% NaCl enriched) develop hypertension (80mmHg higher systolic blood pressure), proteinuria (75% higher), and 4-fold higher glomerular injury compared to vehicle-treated normal control rat. Sorafenib induced glomerular injury was associated with decreased (20-80% lower) renal mRNA expression of key glomerular structural proteins such as nephrin, podocin, synaptopodin, and podoplanin compared to vehicle-treated normal control rat. Renal cortical endothelial-to-mesenchymal transition (EndoMT) was activated in the sorafenib induced glomerular injury model. In the sorafenib treated rats, the renal EndoMT was evident with 20% lower mRNA expression of an endothelial marker WT-1 and 2 to 3-fold higher expression of mesenchymal markers Col III, FSP-1, α-SMA, and vimentin. In conclusion, we developed a rat pre-clinical chronic kidney disease model that manifest glomerular injury. We further demonstrate that the glomerular injury in this model is associated with decreased renal mRNA expression of key glomerular structural proteins and an activated kidney EndoMT.

THE EFFECT OF COMPOUND DM509 ON KIDNEY FIBROSIS IN THE CONDITIONS OF THE EXPERIMENTAL MODEL.

Renal fibrosis is a critical event in the progression of chronic kidney disease (CKD) to end-stage renal disease (ESRD). Unfortunately, there are few options to target renal fibrosis in order to develop novel anti-fibrotic agents that could prevent CKD progression to ESRD. We evaluated the efficacy of a novel dual-acting molecule, DM509, in preventing renal fibrosis using the unilateral ureteral obstruction (UUO) renal fibrosis mouse model. DM509 acts simultaneously as a farnesoid X receptor agonist (FXRA) and a soluble epoxide hydrolase inhibitor (sEHi). In this study, groups of 8-12 weeks old C57BL/6J male mice went through either UUO or sham surgery (n=6/group). Mice were pre-treated with DM509 (10mg/kg/d) or vehicle administered in drinking water one day prior to the UUO surgery. Sham, vehicle and DM509 treatments continued until day 10 and blood and kidney tissue were collected for biochemical, histological, and gene expression analysis at the end of the treatment protocol. The UUO group exhibited kidney dysfunction with elevated blood urea nitrogen (BUN) compared to the sham group (63±7 vs. 34±6 mg/dL). DM509 treatment prevented renal dysfunction as evident from 36% lower BUN level in the DM509 treated UUO mice compared to UUO mice treated with vehicle. Vehicle treated UUO mice demonstrated renal fibrosis with elevated kidney hydroxyproline content (213±11 vs. 49±9 µg/mg protein) and kidney collagen positive area (13±2% vs. 1.1±0.1%) compared to the sham group. We found that DM509 treatment prevented renal fibrosis and DM509 treated mice had 34-66% lower levels of kidney hydroxyproline and collagen positive renal area compared to vehicle-treated UUO mice. In conclusion, our data provide evidence that the novel dual-acting FXRA and a sEHi, DM509, prevented renal dysfunction and renal fibrosis in UUO mouse model.

Upconversion thermometry: a new tool to measure the thermal resistance of nanoparticles.

The rapid evolution in luminescence thermometry in the last few years gradually shifted the research from the fabrication of more sensitive nanoarchitectures towards the use of the technique as a tool for thermal bioimaging and for the unveiling of properties of the thermometers themselves and of their local surroundings, for example to evaluate heat transport at unprecedented small scales. In this work, we demonstrated that KLu(WO4)2:Ho3+,Tm3+ nanoparticles are able to combine controllable heat release and upconversion thermometry permitting to estimate its thermal resistance (in air), a key parameter to model the heat transfer at the nanoscale.

Probiotic strains of lactobacilli and bifidobacteria alter pro- and anti-inflammatory cytokines production in rats with monosodium glutamate-induced obesity.

The aim of this study was to investigate the effect of probiotic strains of Lactobacillus casei IMV B-7280, Bifidobacterium animalis VKL, B. animalis VKB on the pro- and anti-inflammatory cytokines production in Wistar male rats with monosodium glutamate (MSG)-induced obesity. It was established that neonatal administration of MSG to rats leads to increasing levels of the interleukin (IL)-1ß and IL-12, and to decreasing ofthe IL-4, IL-10 and tumor growthfactor (TGF)-ß levels in the bloodserum. After administration of the B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition to obese rats the level of the IL-lP in blood serum wasn't differ from that in the obese rats, that didn't receive of the probiotic bacteria. But there was no statistically signifcant difference comparing with intact rats. The level of the IL-12B p4O in blood serum was decreased under influence of the B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition (18.9%, p < 0.05) and B. animalis VKL (10.5%, p < 0.05) compared with obese rats, not receiving probiotic bacteria, but remained higher than in intact animals. After administration to obese rats ofthe B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition the levels ofthe IL-4, IL- 10 and TGF-ß increased in blood serum comparing with obese rats, not receiving probiotic bacteria. The level of the IL-10 also increased under influence of the B. animalis VKB, and IL-4 - under influence of the L. casei IMVB-7280. Our results suggest that these probiotic bacteria and probiotic composition are able to down-regulation the inflammation in rats with MSG-induced obesity but the strongest anti-inflammatory effects have probiotic composition. The ability of lactobacilli and bifdobacteria to alter the pro- and anti-inflammatory cytokines production, opens perspectives to create new treatments for obesity and metabolic syndrome based on probiotics.

[Low molecular weight regulators of the intracellular insulin signal transduction as a correction method of the insulin resistance in the treatment of type 2 diabetes].

Insulin resistance is the characteristic feature of type 2 diabetes. This condition is manifested in the reduction of peripheral tissues sensitivity to the biological action of insulin and is expressed in the inhibition of cellular glucose absorption and metabolism in response to hormonal stimulation. At the cellular level, disorders which are realized both at the receptor and the postreceptor levels can serve a prerequisite to the formation of insulin resistance and are associated with a change in the amount or dysfunction of major molecular signaling cascade. Thus, the insulin receptor, as well as the other related signaling molecules can be considered as ideal therapeutic targets for the correction of insulin resistance and thus low molecular weight effectors which act on the individual links of insulin signaling cascade may be positioned as a new generation of anti-diabetic agents. This report provides information on the regulators of insulin receptor cascade, main advantages and disadvantages of their impact on biological targets and prospects for their therapeutic use as anti-diabetic drugs.

ESTIMATION OF THE CYTOKINS LEVEL IN EXPERIMENTAL HEPATIC ENCEPHALOPATHY IN RATS.

Connection of the cytokins profile with experimental hepatic encephalopathy (HE) in rats was studied. Investigation was conducted on 20 laboratory rats, in which HE was simulated, using СCl4 injection. The іnterleukins (ІL) level, including, ІL­1ß, ІL­4, ІL­10 and interferon­Î³(IFN­Î³), was determined using immunoassay method with the help of polyclonal antibodies. Enhancement of the proinflammatory ІL­1ß and ІНФ­Î³ content in the rats blood serum while induced HE by 57.9 and 39.5% accordingly (p <0.05), comparing with such in a control and the compensation enhancement of the anti'inflammatory IL­4 and IL­10 level by 34.6 and 75.9% (p < 0.05) was established. The results obtained confirms the cytokins role in pathogenesis of HE, determination of their level constitutes significant criterion for the posttransplantation complications prog' nostication. Inflammation and profile of cytokins constitute the main target in therapy of HE in patients, suffering liver cirrhosis

THE SEARCH OF COMPOUNDS WITH ANTIAGGREGATION ACTIVITY AMONG S-ESTERS OF THIOSULFONIC ACIDS.

According to the current understanding, the hyperactivation of platelets may lead to increased intravascular coagulation and thrombosis. Today a relevant issue is the search for new anti-thrombotic agents that are able to modulate the activity of platelet receptors, thus, influence the processes of activation and aggregation of platelets. The aim of this study was to investigate the effects of newly synthesized thiosulfonate derivatives on platelet aggregation. The activity of the compounds was tested in vitro using platelet-rich plasma. As a result of the screening test, structural formulas of four agents with high antiaggregative activity were established. These compounds inhibited ADP- and collagen-induced platelet aggregation in a dose-dependent manner. Two of these compounds were shown to be more effective inhibitors of aggregation induced by ADP (IC50 - 8-10 µM), as well as collagen (IC50 - 1.5-2.0 µM).

[CARBOHYDRATE METABOLISM IN TYPE 1 DIABETIC RATS UNDER THE CONDITIONS OF THE KIDNEY BEAN PODS AQUEOUS EXTRACT APPLICATION].

The influence of the aqueous pods extract of kidney bean (Phaseolus vulgaris) on indicators of carbohydrate metabolism under the condition of experimental type 1 diabetes in rats was studied. It was shown that long-term oral administration of the extract at a dose of 200 mg/kg to rats leads to the decreasing of blood glucose and glycosylated hemoglobin in the background of chronic hypoinsulinemia conditions. The use of studied extract led to an increase of glycogen synthase activity in rat muscle cells and hexokinase activity in rat liver cells under the conditions of type 1 diabetes. It was estimated that administration of the aqueous extract to control rats and animals with studied model of diabetes increases GLUT-4 protein content in muscle tissue. Thus, the mechanisms of P. vulgaris hypoglycemic action can be related with the ability of the particular phytoconstituents directly effect on key intracellular elements of insulin target tissues carbohydrate metabolism under the conditions of type 1 diabetes.

Novel Anthraquinone-based Derivatives as Potent Inhibitors for Receptor Tyrosine Kinases.

The influence of new derivatives of 9,10-anthraquinone with benzoylthiourea, thiazole, triazole and amino acid fragments on the activity of membrane-associated tyrosine kinases was investigated. Inhibitors of protein tyrosine kinase activity of the membrane fraction, as promising agents to search for new potential anticancer agents among the studied compounds, were discovered.

Proteasomal activity in brain tissue following ischemic stroke in Wistar rats.

Functional as well as structural reorganization of brain tissues takes place in the surrounding and remotes brain areas after focal ischemic lesions. In particular, reactive or regenerative processes have been described to occur in the infarction areas and the contralateral hemisphere. Experiments were performed on 63 rats, divided into 3 groups (each consisted of 21 animals): sham operated, short-term occlusion of the right middle cerebral artery (MCAO) group, and long-term MCAO group. We have studied changes in proteasome proteolysis during transient occlusion of the middle cerebral artery using method of Koizumi J., duration 2 and 60 min and made the comparison between changes in different types of proteasome activity and severity of ischemic injury and showed three types of decrease inproteolytic activity (trypsin-, chymotrypsin-like, peptidylglutamyl peptide-hydrolyzing) in the brain tissues. Chymotrypsin-like activity of ischemic areas of the brain for short-term MCAO decreased 4.1 times compared with controls (P > 0.05), for long-term MCAO decreased 5.8 times compared with controls (P < 0.05). Trypsin-like activity of ischemic areas of brain for short-term MCAO decreased 7.1 times compared with controls (P > 0.05), for long-term MCAO decreased 12.5 times compared with controls (P < 0.05). PGPH activity of ischemic areas for short-term MCAO decreased 8 times compared with controls (P > 0.05), for long-term MCAO decreased 2.8 times compared with controls (P < 0.05). The similar dynamics was observed also in the penumbra and the core zone of the brain at 6 h of reperfusion, in the long run there is no significant difference between the core and contralateral zones. Our results suggest that proteasome activity may play also a role in contralateral cortical plasticity occurring after focal cerebral ischemia.

[FUNCTIONING PROTEASES IN THE ESOPHAGUS MUCOSA AFTER CHEMICAL BURNS].

The main result of esophagus burn is the formation of scars, that caused by excessive synthesis of collagen and changes the balance of metalloproteinases and their tissue inhibitors. It was studied the activity of proteolytic enzymes, participation of MMP (metalloproteinase) and their tissue inhibitors (TIMP) in alkali burns of the esophagus 1st and 2nd degrees. We have shown a significant increase of TIMP level in homogenate after alkali burns of the esophagus (an average of 31-56% depend on of burn degree). We observed a reduced activity of serine proteinase after 1st degree burns on 15th, 21st day 35 and 18% respectively, after burns 2nd degree on 15th, 21st day 54 and 50%. The decrease of activity MMP after 1st degree burns on 15th and 21st day 30, 19%, respectively, in conditions of chemical burns 2nd degree on 15th and 21st day 30, 37%. These data may indicate the development of scarring after burn simulation of 2nd degree. Further investigation of the MMP and TIMP in the process of wound healing can be useful in creating effective approaches to prevent formation of post scarring of the esophagus.

[Optimization of anticoagulation therapy for venous thromboembolism]. / Optimizatsiia antikoaguliantnoi terapii venoznogo tromboémbolizma.

OBJECTIVE: To optimize anticoagulation therapy for venous thromboembolism by means of dabigatran etexilate. MATERIAL AND METHODS: From 2006 to 2012, within the framework of the international trials RE-COVER and RE-COVER II aimed at evaluating efficacy and safety of dabigatran etexilate compared to warfarin, a total of 95 patients meeting the inclusion and exclusion criteria were enrolled in our study. As part of the RE-COVER trial, we carried out analysis of comprehensive examination and treatment of 55 patients with venous thromboembolism (VTE), who were randomly divided into two groups. Group I (Control Group) consisted of 30 patients receiving initial therapy with heparin for seven days followed by taking warfarin for six months. Group II (Study Group) comprised 25 patients taking dabigatran etexilate instead of warfarin. RESULTS: There were no cases of recurrent VTE in the Control Group, and one (4%) patient of the Study Group was found to have a relapse of the disease owing to resistance to anticoagulation therapy and congenital thrombophilia. Undesirable events of anticoagulation therapy developed in 20% of the Control Group patients an in 16% of the Study Group patients. In two Control Group patients and one Study Group patient anticoagulation therapy was discontinued due to the development of complications. After 2 years, 36.7% of the Control Group patients and 40% of the Study Group patients had no manifestations of chronic venous insufficiency (CVI). The degree of CVI was similar in the both groups. CONCLUSION: Dabigatran proved non-inferior to warfarin regarding efficacy, possessing, however, a series of advantages: it has a predictable anticoagulant effect, requires neither monitoring of the haemostasis system, nor dose adjustment, and is administered at standard dosages.

[Frequency of allele polymorphism of immune proteasome catalytic subunits in patients with ischemic stroke].

Frequency of allelic variants of proteasome subunits genes LMP2 (Arg60 --> His) and PSMA6 were determined in patients with ischemic stroke using real-time PCR. Allelic variants of PSMA6 were disposed in the next manner: C/C - 80.2%, C/G - 19.8%, G/G--were not (in control) and C/C - 75.5%, C/G - 21.4%, G/G - 3.1% (P = 0.22) in patients with IS. It was shown that distribution of LMP2 allelic variants was the following: Arg/Arg - 53.3%, Arg/His - 43.5%, His/His - 6.7% in control and Arg/Arg - 55.9%, Arg/His - 34.3%, His/His - 9.8% in IS group (P > 0.05). The data show that LMP2 and PSMA6 gene polymorphism is not a risk factor of ischemic stroke in Ukrainian population.

[Advantages and disadvantages of warfarin and pradaxa therapy for venous thromboembolism].

An analysis of complex examination of 110 patients with venous thromboembolism was made. The patients were separated into 2 groups. The first group included 60 patients, who had the start heparin therapy during 7 days with the following 6-month warfarin therapy. Warfarin was substituted by pradaxa (dabigatran) for 50 patients of the second group. The efficacy of pradaxa could be compared with warfarin. However, pradaxa had a number of advantages such as the predictable anticoagulant effect, standard dosages. This medicine is more predictable and doesn't require a control of homeostasis and an adjustment of drug dosage.

[Development of a prediabetic state under chronic alcohol intoxication].

We investigated the changes in key parameters of carbohydrate and lipid metabolism, which correspond to the clinical picture that accompanies the development of prediabetic condition on the background of chronic alcohol intoxication. From the analysis of glycemic curves obtained during the insulin-glucose test, a speed of glucose uptake by peripheral tissues increased at the 1st day (1.5 fold) and the third day (1.3 fold) of administration of alcohol solution. At the later periods, at 7 and 11 days of ethanol administration, a decreased rate of glucose uptake in animals with chronic alcohol intoxication was detected. We also detected an increased content of serotonin in the blood serum and a decreased (1.2 fold) serotonin content in rat brain during the whole period of development of chronic alcohol intoxication.

[Insulin receptor expression in subcellular fractions of muscular and adipose tissue as the factor of the tissue insulin resistance development in rats under conditions of the high-energy diet].

Nowadays the problem of insulin resistance, which has close cause-effect relations with obesity, diabetes mellitus type 2, metabolic syndrome, etc., is of urgent importance in medicine. We have revealed bidirectional changes of the IR content in crude membrane and cytosol of the adipose tissue cells in rats under conditions of the long-term high-energy diet. It is possible that reduction of the IR content in the adipose tissue cells has been predetermined by the disruption of lipid bilayer of adipocytes as a result of peroxidation processes activation. Increase in the IR content in the cytosol of cells of this tissue may indicate the activation of synthesis of this protein; however, it is possible that the IR translocation process disorder occurs due to the damage of plasma membrane, preventing the transfer of newly synthesized molecules of the receptor to the membrane and causing their accumulation in cytosol. The obtained results show that the tissues react to the long-term consumption of high-energy food in different ways. Thus, the content of insulin receptors in the plasma membrane of the muscle tissue cells increases, and, on the contrary, it decreases in adipose tissue cells. Such results may indicate that IR development at the late period of the experiment is likely the result of the adipose tissue cells disfunction. The obtained data may be of high significance in understanding the mechanism of the IR development under conditions of the long-term consumption of the high-energy food.

[Comparative effectiveness and safety of oral anticoagulants in treatment of acute thromboses of deep veins of the lower extremities].

An analysis of complex examination and treatment of 101 patients with venous thromboembolism was made. The patients were divided into 3 groups. The first group consisted of 46 patients who had starting therapy during 7 days with heparin followed by taking phenylene during 6 months. The second group included 30 patients who were given warfarin instead of phenylene, 25 patients of the third group were treated with dabigatran. In the first group there were 21.7% of relapses, in the second group there were no relapses. In the third group there was 1 (4%) patient with a relapse of the disease due to congenital thrombophilia. Complications of anticoagulation therapy were noted in 23.9% of the first group patients, 20%--in the second group, and 16% of the third group patients.