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Indexes of nitric oxide system in experimental antiphospholipid syndrome
O. Z. Yaremchuk, K. A. Posokhova, І. P. Kuzmak,
M. I. Kulitska, I. М. Klishch, M. M. Korda
I. Horbachevsky Ternopil National Medical University, Ukraine;
e-mail: yaremchuk@tdmu.edu.ua
Received: 11 November 2019; Accepted: 21 January 2020
Antiphospholipid syndrome (APS) is an autoimmune disease characterized by the presence of antibodies to negatively charged membrane phospholipids (aPL). Endothelial dysfunction is one of the most dangerous APS manifestations followed by thrombosis, placental insufficiency and often foetal death due to circulatory disorders in placenta blood vessels. It is established that synthesis and bioavailability of nitric oxide (NO) in the endothelium are impaired at APS, but the role of NO system in pregnancy failure at this pathology remains ambiguous. The aim of this research was to estimate the indexes of the nitric oxide system in animals with an experimental antiphospholipid syndrome before pregnancy and on the 18th day of pregnancy, without treatment and under treatment with nitric oxide synthesis modulators (L-arginine and aminoguanidine). In the blood serum and liver of the BALB/c mice with experimental APS, the content of eNOS and iNOS– by ELISA and the level of NO2– and NO3– with the use of Gris reagent were determined before pregnancy and on the 18th day of pregnancy. The data obtained indicate the relative inefficient NO production by eNOS and NO hyperproduction by iNOS in the blood serum and liver of mice in the pathogenesis of experimental APS. Thus, in mice with APS before pregnancy and on the 18th day of the pregnancy, the eNOS content and NO2– level were decreased while the iNOS content and NO3– level were increased compared to the indexes in the control animal group. L-arginine administration to the animals with APS at the follow-up periods resulted in an increased eNOS content and NO2–, NO3– levels in blood serum and liver with the simultaneous decrease in iNOS content in the liver as compared to indexes in untreated mice with APS. The combined use of L-arginine and selective iNOS inhibitor aminoguanidine caused a significant increase in eNOS content and a decrease in iNOS content followed by normalization of NO2– and NO3– levels in blood and liver of mice with experimental APS before pregnancy and on the 18th day of pregnancy compared to untreated mice with APS.
Methotrexate effect on biochemical indices of psoriasis patients depends on MTHFR gene polymorphism
O. M. Fedota1, L. V. Roschenyuk2,3, T. V. Tyzhnenko1,
N. G. Puzik1,3, V. M. Vorontsov1, P. P. Ryzhko1
1V.N. Karazin Kharkiv National University, Ukraine;
2Kharkiv Regional Clinical Skin and Venereal Diseases Dispensary №1, Ukraine;
3Kharkiv National Medical University, Ukraine;
e-mail: tyzhnenko@ukr.net
Received: 13 June 2019; Accepted: 29 November 2019
Methotrexate (MTX) is the immunosuppressive anti-inflammatory drug and the antagonist of the enzyme dihydrofolate reductase. Pharmacogenomic studies and clinical evidences suggest that altered response to MTX in patients with different diseases is associated with polymorphisms of genes that regulate folate metabolism. The purpose of the article was to analyze the methotrexate effect on the biochemical indices of psoriasis patients depending on methylenetetrahydrofolate reductase gene (MTHFR) polymorphisms. Effects of two single-nucleotide polymorphisms, C677T and A1298C, were studied. An increase of alanine aminotransferase and aspartate aminotransferase activity above the normal level in the patients with both MTHFR gene polymorphisms after methotrexate intake was observed. In patients with CC, TT, CT genotypes for C677T polymorphism and AA genotype for A1298C polymorphism of MTHFR gene, significant differences in alpha-amylase activity before and after treatment with methotrexate were detected. Analysis of the biochemical indices of patients with arthropathic and vulgaris psoriasis showed that the positive effect of MTX treatment could be associated with wild-type alleles in both polymorphisms of MTHFR gene, while the ineffectiveness of methotrexate was associated with the dihеterozygous genotype. The largest number of smokers was found within the CTAA genotype group (37.5%), while no smokers were observed within TTAA patients and most of CCAA patients. The data obtained testify the utility of the individual approach to the psoriasis patients therapy taking into account genetic background.
Profiling of metabolic biomarkers in the serum of prostate cancer patients
F. Ali1, S. Akram1, S. Niaz1,2, N. Wajid1
1Institute of Molecular Biology and Biotechnology (IMBB) & Centre for Research In Molecular Medicine (CRIMM), The University of Lahore, Raiwind Road Lahore, Pakistan;
2Social Security Hospital Multan Chungi, Multan Road, Lahore;
e-mail: Fatima.ali@imbb.uol.edu.pk; fatemei.ali@gmail.com
Received: 26 July 2019; Accepted: 29 November 2019
Prostate cancer (PCa) is the major cause of the death of men population globally. Multiple factors are involved in the initiation and progression of PCa. This study aimed to evaluate different metabolic parameters in the serum of PCa patients. Males of 50 years and above age with the recent diagnosis of PCa (digital rectal examination, and elevated serum prostate-specific antigen (PSA) level) were included in the study. Glucose and serum electrolytes level, lactate dehydrogenase activity, parameters of lipid metabolism and liver and kidney functioning were measured on a fully automated analyzer using standard reagent kits. Oxidative stress was evaluated by measuring MDA, CAT, GSH, and SOD in serum. Detection of C-reactive protein (CRP), insulin-like growth factor (IGF-1) and vascular endothelial growth factor (VEGF) was performed by immunoassay. It was shown that serum glucose and HDL levels were lower while total cholesterol, LDL and triglyceride levels were significantly higher in PCa group than in the control group. PCa patients had an elevated level of liver and kidney functional markers. Comparison of the oxidative stress markers in patient and control groups showed significant difference. It was detected that serum levels of CRP, IGF-1 and VEGF were significantly higher in PCa group, compared the control to group (P < 0.05). Low level of glucose and dyslipidemia indices in prostate cancer patients indicated metabolic changes and demonstrated the importance of multiple parameters analysis (free PSA, dyslipidemia, VEGF, IGF-1, CRP, and oxidative stress markers) for early PCa diagnostics.
Influence of Tl(+) on the Ca(2+) and Na(+) movement across rat neonatal cardiomyocytes and rat heart mitochondria membranes
S. M. Korotkov, V. P. Nesterov, G. B. Belostotskaya,
I. V. Brailovskaya, A. V. Novozhilov, C. V. Sobol
Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russian Federation;
e-mail: korotkov@SK1645.spb.edu
Received: 05 September 2019; Accepted: 29 November 2019
Thallium is known to produce one of the most complex and serious patterns of toxicity, involving a wide range of human organs and tissues. The toxic impact on biologic organisms is linked especially to the ability of Tl+ to disturb calcium homeostasis and to permeate easily the inner mitochondrial membrane (IMM). The aim of this work was to study the effects of Tl+ on intracellular Ca2+ dynamics in rat neonatal cardiomyocytes as well as on sodium penetrability of the IMM and Tl+-induced mitochondrial permeability transition pore (MPTP) opening in isolated Ca2+-loaded rat heart mitochondria (RHM). The use of the fluorescent calcium indicator Fura 2 AM showed that Tl+ induced calcium influx across the plasmatic membrane, resulting in calcium ([Ca2+]i) increase in the cytoplasm. This increase was even more pronounced in experiments with accelerating of Tl+-transmembrane fluxes by nonactin. It was nevertheless abolished by the removal of extracellular Ca2+ ions, but was not inhibited by a calcium-channel blocker (nifedipine). Tl+ did not release calcium from the intracellular stores. Tl+ potentiated sodium permeability of the IMM because swelling of nonenergized RHM in medium containing TlNO3 and NaNO3 was enhanced at high Tl+ concentration. The calcium load of RHM induced MPTP opening which was accompanied by the increase of the swelling as well as the decrease of the inner membrane potential and of state 40 (basal) and state 3UDNP (2,4-dinitrophenol-uncoupled) respiration. These effects of Tl+ were suppressed by MPTP inhibitors (cyclosporine A, ADP and n-ethylmaleimide). The data obtained showed that Tl+-stimulated influx of extracellular calcium into cardiomyocytes could cause calcium and sodium RHM overload, which lead to the MPTP opening, thus determining the sensitivity of heart muscle to thallium intoxication.
Plasminogen modulates formation and release of platelet angiogenic regulators
A. A. Tykhomyrov, D. D. Zhernosekov, T. V. Grinenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: artem_tykhomyrov@ukr.net
Received: 19 July 2019; Accepted: 29 November 2019
Platelets store, produce and release a variety of angiogenesis regulators, which can contribute to both normal tissue repair and angiopathy-associated pathologies. Plasminogen has been earlier shown to regulate some platelet functions, but if it is able to modulate angiogenic capacities of platelets is still poorly studied. Thus, the aim of the present study was to evaluate the effects of different plasminogen forms on the formation and secretion of angiogenic protein regulators by platelets. Human washed platelets were obtained by gel-filtration on Sepharose-2B. The levels of P-selectin (CD-62P) exposed on the plasma membrane of untreated and activated platelets was monitored by flow cytometry. Secretion of platelet-derived vascular endothelial growth factor (VEGF) as well as plasminogen fragmentation and angiostatin formation by intact platelets and platelet plasma membranes were analyzed by immunoblotting. It was shown that thrombin or collagen exposure resulted in enhanced P-selectin surface expression by platelets, while Lys-form of plasminogen reduced agonist-induced platelet secretion. Lys-plasminogen, but not Glu-form, inhibited agonist-induced VEGF release from platelets. Activation of platelets significantly accelerated plasminogen cleavage and angiostatin formation. Anti-actin antibodies inhibited plasminogen fragmentation during incubation with platelet plasma membranes indicating surface-exposed actin participation in plasminogen conversion to angiostatins. The present study uncovers a novel function of plasminogen to limit angiogenic potential of platelets via angiostatin formation and inhibition of VEGF secretion.
Inhibition of Na(+),K(+)-ATPase and activation of myosin ATPase by calix[4]arene C-107 cause stimulation of isolated smooth muscle contractile activity
T. O. Veklich1, R. D. Labyntseva1, O. A. Shkrabak1, O. V. Tsymbalyuk2,
R. V. Rodik3, V. I. Kalchenko3, S. O. Kosterin1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Institute of High Technologies, Taras Shevchenko National University of Kyiv, Ukraine;
3Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: veklich@biochem.kiev.ua; otsymbal@bigmir.net; manli@ioch.kiev.ua
Received: 04 Jule 2019; Accepted: 29 November 2019
The discovery of compounds that might modify myometrial contractility is an important area of researches. In our previous experiments, we found that some representatives of macrocyclic compounds family – calix[4]arenes – can modify the enzymatic and transport activity of membrane-bound cation-transport ATP hydrolases. The aim of this work was to study and compare the effect of calix[4]arene C-107 on the enzymatic activities of Mg2+-dependent ATPases of the uterine smooth muscle, namely: ouabain-sensitive Na+,K+-ATPase, plasma membrane Ca2+-independent “basal” Mg2+-ATPase, ATPase of the actomyosin complex and myosin subfragment-1, with effect on the contractile activity of the myometrium. It was shown that calix[4]arene C-107 efficiently inhibited myometrium Na+,K+-ATPase (I50 = 54 ± 6 nM) selectively to other ATP-hydrolases of the plasma membrane and simultaneously activated the enzymatic activity of the myosin ATPase of smooth muscles (A50 = 9.6 ± 0.7 μM). Such reciprocal biochemical effects led to the stimulation of the smooth muscle contractile activity that was demonstrated by the tensometric method using different isolated smooth muscles. Calix[4]arene С-107 was shown to stimulate the increase of the tonic component of myometrium contractions induced by oxytocin, as well as contractions of the caecum muscles induced by high-potassium solution or acetylcholine, and to maintain increased tension for a long time. Thus, calix[4]arene C-107 is a prospective compound for enhancing the smooth muscle basal tone and/or contraction in case of hypotonic dysfunctions.
Proline dehydrogenase (PRODH) gene polymorphisms and the risk of schizophrenia in Iranian populations
F. H. Moghadam1, Z. H. A. Mehrabani2, M. Amounajaf3,
S. Rahmanzadeh3, F. Ghasemvand3, A. S. Samghabadi3,
A. Nejadmoghaddam3, E. Omidinia3
1Department of Traditional Medicine, School of Traditional Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
2Department of Clinical Biochemistry and Laboratory Medicine, Tabriz University of Medical Sciences, Tabriz, Iran;
3Enzyme Technology Lab., Genetics & Metabolism Research Group, Pasteur Institute of Iran, Tehran, Iran;
e-mail: saeed_r81@yahoo.com or 2000.spss@gmail.com
Received: 10 August 2019; Accepted: 29 November 2019
Schizophrenia is a highly heritable mental disorder which can be occurred as a result of mutations or single nucleotide polymorphisms (SNPs) in various genes. Proline dehydrogenase (PRODH) gene is one of the most important genes which can be associated with increased risk of schizophrenia in several populations. Here, we considered the effect of PRODH gene polymorphisms on the incidence of schizophrenia in Iranian populations. This study was done using the analysis of 3 SNPs markers, including G1496A, G758A and C1482T. Molecular analysis was performed on 263 schizophrenic patients and 278 healthy individuals (control group). These examinations were executed by PCR-based restriction fragment length polymorphism (RFLP) technique. Statistical analysis was performed by SPSS software (16.0). Our findings showed that G1496A and C1482T polymorphisms in patients were significantly higher than controls and there were meaningful correlations between the occurrence of these polymorphisms and schizophrenia in the population (P < 0.001). However, there was no significant relationship between G758A in the PRODH gene and schizophrenia. Haplotype analysis showed that AAT, AAC and GAT blocks (variation alleles are bold) had significant correlations with schizophrenia. PRODH gene can be considered as one of the important genes involved in schizophrenia development among the Iranian population.
Pattern of expression of immune- and stroma-associated genes in blood of mice with experimental B16 melanoma
G. V. Gerashchenko, I. M. Vagina, Yu. V. Vagin, V. I. Kashuba
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
e-mail: g.v.gerashchenko@imbg.org.ua
Received: 30 May 2019; Accepted: 29 November 2019
The interaction between malignant and stromal cells represents a major cross-talk pathway upon carcinogenesis. Cellular elements of the reactive tumor stroma are a heterogeneous population which are represented specifically by cancer-associated fibroblasts (CAFs) and tumor-associated macrophages (TAM). It is not known whether expression of CAF- and TAM-associated genes could be detected in the peripheral blood of cancer patients to monitor a course of disease. The aim of the study was to assess the relative expression (RE) of cancer-related genes in peripheral blood of mice with experimental melanoma. Quantitative PCR was used to determine RE of 15 genes in the blood of C57BL/6j control mice and mice with injected B16 melanoma cells. The Kruskal-Wallis and the Fischer exact tests with correction on multiple comparisons, according to the Benjamini-Hochberg procedure with FDR = 0.2 were used for statistical analysis. Analysis of 15 immune and stromal markers RE showed differentiated expression of several CAF and TAM markers in mice with experimental melanoma in comparison with the control animals. Thus, CAF markers Acta2, Cxcl14, Fap and TAM markers Cd68, Ccl22 and Ccl17 were significantly upregulated, while Cd4, Cd3 were downregulated. This, together with increased expression of Cox-2 suggested a stable immunosuppressive state of mice with experimental melanomas. The results of the study showed that potential markers of cancer-associated fibroblasts and tumor-associated macrophages in peripheral blood of mice with experimental melanoma could be used for non-invasive detection of melanoma cell progression.