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Carnitine effects on serum and pancreas inflammatory response in diabetic rats
Y. Masoumi-Ardakani1, H. Fallah2, B. Shahouzehi3
1Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
2Student Research Committee, School of Medicine,
Kerman University of Medical Sciences, Kerman, Iran;
3Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
e-mail: bshahouzehi@yahoo.com, bshahouzehi@gmail.com
Received: 08 May 2019; Accepted: 18 October 2019
Diabetes is a group of disorders characterized by elevated blood glucose and insulin secretion defect. Previous studies have reported L-carnitine beneficial and hypoglycemic effects in diabetic models. L carnitine anti-inflammatory properties in diabetes were not assessed perfectly, and there is a lack of information about this matter. Therefore, we designed this study and evaluated L-carnitine different doses supplementation on pro-inflammatory cytokines in STZ-induced diabetic rats’ pancreas and serum. We selected 48 male rats (200 ± 10 g) and randomly divided them into six groups (n = 8). Group 1, control; group 2, Diabetic control (DC); groups 3-6, STZ-induced diabetic rats which received L-carnitine different doses as follow; 300, 200, 100 and 50 mg/kg/day by intraperitoneal injection for 5 weeks. When the study ended, serum and pancreas samples were collected and cytokines levels were measured by specific ELISA kits. Our results showed that in diabetic rats, pro-inflammatory cytokines levels were elevated. Two L-carnitine doses 300 and 200 mg/kg/day showed beneficial effects and 300 mg/kg/day showed more effective and significant effects than other doses. The 300 mg/kg significantly reduced IL-1β and IL-6 levels in pancreas and serum. Our data proved the protective effects of intraperitoneal L-carnitine administration against diabetes and inflammation in diabetic rats. Indeed, L-carnitine long term supplementation through the intraperitoneal injection can be considered as a good and safe therapeutic strategy in diabetes.
Effect of probiotic composition on oxidative/antioxidant balance in blood of rats under experimental osteoarthritis
O. Korotkyi, K. Dvorshchenko, A. Vovk, A. Dranitsina, M. Tymoshenko, L. Kot, L. Ostapchenko
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: korotky@ukr.net
Received: 20 June 2019; Accepted: 18 October 2019
Osteoarthritis (OA) is a widespread pathology of the musculoskeletal system. OA may associate with a wide range of disorders, that lead to the development of various strategies on how to prevent and treat the disease. Recent studies discussed interactions between the microbiome and a wide range of pathologies, including OA. In this study, we investigated the effect of probiotic cultures on oxidative/antioxidant balance in blood of rats during OA. Experimental OA was induced by a single injection of sodium monoiodoacetate into the knee joint. A probiotic composition (Multiprobiotic Simbiter®) was administered by peroral catheter once per day for 14 days. We investigated the next parameters: expression of Nos2 gene in the blood, superoxide dismutase activity, catalase activity, glutathione peroxidase activity, glutathione transferase activity, glutathione reductase activity, contents of superoxide, hydrogen peroxide, TBA-reactive compounds, oxidized and reduced glutathione in the serum of the blood. Monoiodoacetate-induced OA caused significant changes on oxidative/antioxidant balance in the blood of rats: increasing of the contents of superoxide anion radical, hydrogen peroxide, thiobarbituric acid-reactive compounds, oxidized glutathione, upregulating of Nos2 expression, increasing of catalase activity; conversely, superoxide dismutase activity, glutathione peroxidize activity, glutathione transferase activity, glutathione reductase activity, the content of reduced glutathione were significantly decreased, compared to control group. Administration of probiotics to animals with OA led to positive changes in the studied parameters approaching the values of control group (some of them were statistically significant).
Fibrinolysis regulation by platelets retaining plasminogen and tissue-type plasminogen activator on their surface
T. Grinenko, О. Yusova, O. Revka, I. Patalakh, T. Yatsenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: sedrickedel@gmail.com
Received: 22 July 2019; Accepted: 18 October 2019
Platelets play a key role in hemostasis as cofactors of thrombin generation, fibrin polymerization centers, and initiators of clot retraction, while their ability to modulate clot dissolution remains less understood. The aim of this study was to investigate the interaction of plasminogen and tissue plasminogen activator with native and activated platelets, to determine the amount of plasmin generated by various activators in the presence of platelets, and the ability of platelets to modulate the rate of polymer fibrin hydrolysis. Spectrometric and immunofluorometric methods were used in the study. It was shown that intact circulating platelets carry a small amount of plasminogen on their surface, whereas thrombin-induced activation led to the exposure of plasminogen-binding sites on their plasma membrane. Activated platelets stimulated plasminogen activation by tissue plasminogen activator, urokinase, and streptokinase. Components of prothrombin complex enhanced plasminogen activation by tissue plasminogen activator on the surface of activated platelets. Model system with desAB-fibrin revealed the ability of platelets to stimulate fibrinolysis. These results suggest that the regulation of fibrinolysis by platelets is provided by the binding of plasminogen and plasminogen activators on their surface, acceleration of plasmin generation and, consequently, acceleration of the onset of fibrin lysis and reducing of the clot lifetime, which is important to maintain hemostatic balance.
Comparative analysis of the influence of chlorine and fluorine anions on the fibrin polymerization
L. V. Pyrogova1, G. K. Bereznitsky1, G. K. Gogolinskaya1, T. M. Platonova1,
I. M. Kolesnikova1, O. O. Masenko1, R. Yu. Marunich1, P. Yu. Tsap1,
Yu. V. Ushenin2, Y. M. Makogonenko1, E. V. Lugovskoi1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2V.E. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ymakogonenko@gmail.com
Received: 25 April 2019; Accepted: 18 October 2019
The effect of NaCl and NaF salts in the range of 0.1-0.225 M concentrations on individual stages of fibrin polymerization was investigated, namely: the rate of fibrinogen activation by thrombin, the rate of protofibrils formation, the rate of lateral association of protofibrils, and the maximum clot absorbance value at 350 nm. It was found that the chlorine and fluorine anions equally inhibit the rate of formation of fibrin from fibrinogen and the formation of protofibrils activated with thrombin. Chlorine anions were shown to be significantly more effective than fluorine anions, inhibiting the rate of lateral association and maximal clot absorbance level from fibrin desA and desAB. A component of the inhibitory action of chlorine anions, not related to the ionic strength of the solution, was identified and its effect on the individual polymerization steps was shown. Chlorine anions were found to bind to a fibrin clot. Using the surface plasmon resonance method and fibrin-specific mAb FnI-3c, it was established that the rate of exposure of neoantigenic determinants of mAb in the hinge regions of the fibrinogen molecule during its transformation into fibrin under the action of thrombin is inhibited by chlorine anions in correlation with inhibition of the protofibril lateral association rate. It has been suggested that the inhibitory effect of chlorine anions consists of an ionic component and a component that blocks the conformational mobility of the molecule by the chlorine anions binding to its hinge regions and polymerization sites.
Analysis of Aurora kinases genes expression points on their distinct roles in prostate cancer development
O. Mankovska1, G. Gerashchenko1, E. Rozenberg1, E. Stakhovsky2,
O. Kononenko2, Yu. Bondarenko3, V. Kashuba1,4
1Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
2National Cancer Institute, Ministry of Health of Ukraine, Kyiv;
3Institute of Urology National Academy of Medical Sciences of Ukraine, Kyiv;
4Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Stockholm, Sweden;
e-mail: mankovsska@gmail.com
Received: 28 May 2019; Accepted: 18 October 2019
Aurora kinases A and B play a crucial role in the regulation of mitosis, while Aurora C controls meiotic division. These proteins showed controversial behavior upon the development of epithelial tumors. Our aim was to examine if there are any differences in expression of Aurora kinases genes in malignant and non-malignant tumors and non-tumor tissues; to compare their expression with clinical characteristics of patients and expression of other prostate cancer-associated genes. Quantitative RT-PCR was used to determine Aurora A-C genes expression in 33 prostate adenocarcinomas (T), paired conventionally normal tissues (N), and 17 adenomas (A). Relative expression values (RE) for genes studied were estimated using 2-ΔCt and 2-ΔΔCt method. The Kruskal-Wallis with correction on multiple comparisons, according to the Benjamini-Hochberg procedure with FDR = 0.2, Dunn-Bonferroni post hoc test and Spearman rank correlation analysis were used for statistical analysis. As turned out, RE values for AURKA were found to be significantly lower in samples of T group in comparison with A group. Moreover, significant up-regulation of AURKC expression levels were detected for T 3-4 stage compared to T 1-2 stage. RE values of AURKC in T group were positively correlated with the tumor stage, while AURKB RE demonstrated a negative correlation with the tumor stage. We also found significant correlations between AURK genes expression levels and prostate cancer-associated genes in T group. We suppose that all these data point to probable involvement of Aurora kinases’ genes in prostate carcinogenesis.
Dependence of the mitochondrial adaptive capacity of hepatocytes on the oxidative substrates availability
H. M. Mazur, V. M. Merlavsky, B. O. Manko, V. V. Manko
Ivan Franko National University of Lviv, Ukraine;
e-mail: Volodymyr.Manko@lnu.edu.ua
Received: 11 February 2019; Accepted: 18 October 2019
The ability of the mitochondria to compensate for energy expenditure of cells largely depends on the availability of the oxidative substrates, transported across the intact plasma membrane with molecular carriers of limited affinity. The aim of this study was to investigate the dependence of adaptive respiratory responses of mitochondria of intact hepatocytes on the oxidative substrates. Basal and FCCP-stimulated respiration rates were determined with Clark electrode. After 15-minute incubation in the medium with the oxidative substrates or their combinations (glutamine, pyruvate, succinate, monomethyl succinate, α-ketoglutarate, dimethyl-α-ketoglutarate (2 mM) or glucose (10 mM)), isolated hepatocytes were added into the respiratory chamber. FCCP concentration was 0.25, 0.5 and 1 μM. The adaptive capacity of mitochondria was characterized by the maximal uncoupled respiration rate (the highest respiration rate among all tested FCCP concentrations), the optimal FCCP concentration (the concentration at which the maximal rate is achieved) and the area under the curve (AUC) of the dependence of the uncoupled respiration rate on FCCP concentration. The adaptive capacity of mitochondria, evaluated by AUC, increases in this order of substrates: glucose (0.063 r.u.), endogenous substrates (0.067 r.u.), glutamine (0.092 r.u.), pyruvate (0.113 r.u.), α-ketoglutarate (0.113 r.u.), succinate (0.152 r.u.), dimethyl-α -ketoglutarate (0.156 r.u.), and monomethyl succinate (0.172 r.u.). The adaptive capacity of mitochondria of hepatocytes seems to be partly dependent on plasma membrane transporters affinities (Km) to the oxidative substrates. The presence of glucose in the medium does not improve the adaptive capacity of hepatic mitochondria.
The effect of quercetin on oxidative stress markers and mitochondrial permeability transition in the heart of rats with type 2 diabetes
N. I. Gorbenko1, O. Yu. Borikov2, O. V. Ivanova1, E. V. Taran1,
Т. S. Litvinova1, T. V. Kiprych1, A. S. Shalamai3
1V. Danilevsky Institute of Endocrine Pathology Problems, National Academy of Medical Sciences of Ukraine, Kharkiv;
2V. N. Karazin Kharkiv National University, Ukraine;
3PJSC SIC “Borshchahivskiy Chemical-Pharmaceutical Plant”, Kyiv, Ukraine;
е-mail: Gorbenkonat58@ukr.net
Received: 24 June 2019; Accepted: 13 August 2019
Increasing evidence suggests that oxidative stress and induction of mitochondrial permeability transition in cardiomyocytes are linked to tissue damage and the development of diabetic cardiovascular complications. The aim of this study was to assess the effects of quercetin (Q) on oxidative stress and mitochondrial permeability transition in the heart of rats with type 2 diabetes mellitus (DM). Type 2 DM was induced in 12-week-old male Wistar rats by intraperitoneal injections of 25 mg/kg streptozotocin twice per week followed by a high-fat diet during four weeks. The rats were divided into three groups: control intact group (C, n = 8), untreated diabetic group (Diabetes, n = 8) and diabetic rats treated with Q (50 mg/kg/day per os for 8 weeks) after diabetes induction (Diabetes+Q, n = 8). Administration of Q increased insulin sensitivity and normalized the functional state of cardiac mitochondria due to increased aconitase and succinate dehydrogenase activities in rats with type 2 DM. Q also ameliorated oxidative stress, decreasing the level of advanced oxidation protein products and increasing the activity of thioredoxin-reductase in heart mitochondria of diabetic rats. In addition, Ca2+-induced opening of the mitochondrial permeability transition pore was significantly inhibited in diabetic rats treated with Q in comparison with the untreated diabetic group. These data demonstrate that Q can protect against oxidative stress, mitochondrial permeability transition induction and mitochondrial dysfunction in cardiomyocytes of diabetic rats. We suggest that the use of Q may contribute to the amelioration of cardiovascular risk in type 2 DM.
Brownian motion, electrophoresis, chromatography, and macromolecular chemistry: how it all unites Nobel laureates of the first half of the 20th century – T. Svedberg, A. Tiselius, R. Synge and H. Staudinger
M. V. Grigorieva, V. M. Danilova, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kуiv;
e-mail: mvgrigorieva@biochem.kiev.ua
Received: 31 July 2019; Accepted: 13 August 2019
It is hard to imagine how chemistry, biology and medicine would develop without such techniques as ultracentrifugation, electrophoresis and chromatography. At present innovative hi-tech laboratory ultracentrifuges are widely used in various fields of fundamental science and practice, including colloid chemistry, biochemical analysis, virology, clinical diagnostics, pharmacy, nanotechnology, to name a few. Electrophoresis enables to detect protein abnormalities with high probability, and, therefore, has wide application for the diagnosis of infectious-inflammatory diseases, genetic and immune disorders, malignant tumors and others. Chromatography is widely used for biochemical research and analytical detection and control of drugs and food. But where did these methods, which have had a significant impact on the development of various fields of scientific and economic activity, come from? Who were the pioneers in this field and whose work influenced the formation of the next generation of researchers? These are the questions we address in this article.
Alpha-fetoprotein as a biochemical diagnostic and prognostic marker for prolonged jaundice in newborns
O. H. Mazur, O. S. Yablon, O. S. Rubina,
M. M. Puhach, A. P. Konoplitska
National Pirogov Memorial Medical University, Vinnytsia, Ukraine;
e-mail: alena523@ukr.net
Received: 14 January 2019; Accepted: 13 August 2019
Prolonged jaundice of newborns is a common pathology during the neonatal period. Recently, there has been a tendency toward an increased number of newborns with prolonged jaundice with duration longer than 14 days of life. According to the polyetiologic nature of neonatal jaundice, it is necessary to investigate new diagnostic signs that can predict the development of prolonged jaundice in newborns and, allow identification of new methods of differential diagnosis of neonatal jaundice, as well as decrease the frequency of this pathology. The parameters of serum alpha-fetoprotein (AFP) content in newborns with prolonged jaundice were studied. The content of AFP in blood serum was 1.7 times higher in newborns with prolonged jaundice than in newborns with jaundice for up to 14 days of life: Me = 671.1 [Q1 = 479.9; Q3 = 815.03] ng/ml and Me = 401.0 [Q1 = 284.9; Q3 = 684.0] ng/ml, respectively (P < 0.05). The content of blood serum AFP in newborns with prolonged jaundice was higher for total serum bilirubin greater than 250 μmole/l: Me = 626.2 [Q1 = 454.7; Q3 = 793.2] ng/ml which was confirmed by a strong direct correlation relationship (rxy = 0.64, P < 0.05). The results of the ROC analysis confirmed that AFP ≥ 571.7 ng/ml can identify newborns with prolonged jaundice with a sensitivity of 74.2% and a specificity of 74.5%. The area under the curve (AUC) was 0.870 (95% CI 0.804, 0.937), which confirms the good quality of the AFP model ≥ 571.7 ng/ml.
The role of resistin in the genesis of metabolic disorders in pathological pregnancy
S. O. Ostafiichuk
Ivano-Frankivsk National Medical University, Ukraine;
e-mail: svitlana.ostafijchuk@gmail.com
Received: 12 March 2019; Accepted: 13 August 2019
Pathological gestational weight gain (GWG) is a risk factor for obstetric and perinatal complications. High metabolic activity of adipose tissue and the placenta during pregnancy manifests as an increased production of adipokines that are involved in glucose regulation and insulin sensitivity. The aim of this study was to determine the role of resistin in the genesis of metabolic disorders in pathological GWG pregnancies. The 163 pregnant women were examined in the study: 97 (59.5%) had normal, 18 (11.0%) had insufficient and 48 (29.4%) had excessive prepregnancy weight and obesity. GWG was the recommended level in 56 (34.4%), insufficient in 33 (20.2%), and excessive in 74 (45.4%) women. Anthropometry was performed in each trimester of pregnancy, the weight gain was measured, and the percentage of body fat mass, concentrations of resistin, glucose, insulin, and the HOMA-IR were evaluated. Positive associations were found between hyperresistinemia in the second trimester of pregnancy, and subsequent weight gain (r = 0.27, P = 0.0006), percentage of body fat mass (r = 0.93, P = 0.000) and insulin resistance (r = 0.89, P = 0.000) in late pregnancy; these associations were especially evident in excessive GWG. Determination of predictors of insulin resistance, associated with endocrine activity of adipose tissue, such as the adipokine resistin, in the second trimester of pregnancy may help to predict the severity of metabolic shifts during pregnancy and the risk of developing obstetric and perinatal complications.