Y. Stohnii, M. Ryzhykova, A. Rebriev, M. Kuchma, R. Marunych, V. Chernyshenko, V. A. Shablii, N. M. Lypova, O. Slominskyi, L. Garmanchuk, T. Platonova, S. Komisarenko
The fibrinogen molecule contains multiple binding motifs for different types of cellular receptors, acting as a molecular link between coagulation and cell adhesion. In this study we generated a truncated form of the fibrinogen molecule lacking the Bβ1-42 sequence by site-specific proteolysis and evaluated the role of the fragment in adhesive capabilities of platelets, endothelial and cancer cells. Fibrinogen with the removed Bβ1-42 sequence and fibrin without the Bβ15-42 fragment (desβ1-42 fibrinogen and desABβ15-42 fibrin) were obtained by proteolysis using the specific protease from the venom of Echis multisquamatis. The cleaved fragment was purified by HPLC and was identified using MALDI-TOF. ADPand collagen-induced aggregation of washed platelets in the presence of fibrinogen desBβ1-42 was studied using an aggregometer. Proliferation of mice aortic endothelial cells (MAEC) and human umbilical vein endothelial cells (HUVEC) was studied using the fibrin desABβ15-42 as the scaffold. Cell viability was quantified by the MTT test (MAEC). Generation time was calculated for the estimation of proliferative activity of HUVEC. Lung cancer cell line Н1299 was used to evaluate cancer cell motility in vitro using the scratch assay. Direct comparison of cellular behavior in the presence of truncated vs native forms demonstrated attenuated cell adhesion in the presence of fibrinogen desBβ1-42 and fibrin desBβ15-42. The platelet aggregation rate was only slightly decreased in the presence of fibrinogen desBβ1-42 but resulted in 15-20% disaggregation of adhered platelets. We also observed the substantial decrease of generation time of HUVEC and inhibition of viability of MAEC cells grown on scaffolds of a desABβ15-42 matrix. Finally, desBβ1-42 modulated the motility of H1299 cells in vitro and suppressed the wound healing by 20% compared to the full-length fibrinogen. We postulate that fragment 1-42 of the BβN-domain of fibrinogen is not sufficient for platelet aggregation, however it may contribute to platelet clot formation in later stages. at the same time, this fragment may be important for establishing proper cell-to-cell contacts and cell viability of endothelial cells. Also, 1-42 amino acid fragment of the BβN-domain supported the migration of cancer cells suggesting that interactions of fibrinogen with cancer cells could be a target for anticancer therapy. The Bβ1-42 fragment of fibrinogen contributes to efficient intracellular interactions of different types of cells, including platelets, endothelial cells and cancer cells.
{"title":"Aggregation of platelets, proliferation of endothelial cells and motility of cancer cells are mediated by the B?1(15)-42 residue of fibrin(ogen)","authors":"Y. Stohnii, M. Ryzhykova, A. Rebriev, M. Kuchma, R. Marunych, V. Chernyshenko, V. A. Shablii, N. M. Lypova, O. Slominskyi, L. Garmanchuk, T. Platonova, S. Komisarenko","doi":"10.15407/ubj92.02.072","DOIUrl":"https://doi.org/10.15407/ubj92.02.072","url":null,"abstract":"The fibrinogen molecule contains multiple binding motifs for different types of cellular receptors, acting as a molecular link between coagulation and cell adhesion. In this study we generated a truncated form of the fibrinogen molecule lacking the Bβ1-42 sequence by site-specific proteolysis and evaluated the role of the fragment in adhesive capabilities of platelets, endothelial and cancer cells. Fibrinogen with the removed Bβ1-42 sequence and fibrin without the Bβ15-42 fragment (desβ1-42 fibrinogen and desABβ15-42 fibrin) were obtained by proteolysis using the specific protease from the venom of Echis multisquamatis. The cleaved fragment was purified by HPLC and was identified using MALDI-TOF. ADPand collagen-induced aggregation of washed platelets in the presence of fibrinogen desBβ1-42 was studied using an aggregometer. Proliferation of mice aortic endothelial cells (MAEC) and human umbilical vein endothelial cells (HUVEC) was studied using the fibrin desABβ15-42 as the scaffold. Cell viability was quantified by the MTT test (MAEC). Generation time was calculated for the estimation of proliferative activity of HUVEC. Lung cancer cell line Н1299 was used to evaluate cancer cell motility in vitro using the scratch assay. Direct comparison of cellular behavior in the presence of truncated vs native forms demonstrated attenuated cell adhesion in the presence of fibrinogen desBβ1-42 and fibrin desBβ15-42. The platelet aggregation rate was only slightly decreased in the presence of fibrinogen desBβ1-42 but resulted in 15-20% disaggregation of adhered platelets. We also observed the substantial decrease of generation time of HUVEC and inhibition of viability of MAEC cells grown on scaffolds of a desABβ15-42 matrix. Finally, desBβ1-42 modulated the motility of H1299 cells in vitro and suppressed the wound healing by 20% compared to the full-length fibrinogen. We postulate that fragment 1-42 of the BβN-domain of fibrinogen is not sufficient for platelet aggregation, however it may contribute to platelet clot formation in later stages. at the same time, this fragment may be important for establishing proper cell-to-cell contacts and cell viability of endothelial cells. Also, 1-42 amino acid fragment of the BβN-domain supported the migration of cancer cells suggesting that interactions of fibrinogen with cancer cells could be a target for anticancer therapy. The Bβ1-42 fragment of fibrinogen contributes to efficient intracellular interactions of different types of cells, including platelets, endothelial cells and cancer cells.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"72-84"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43814858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
diabetes is a metabolic disorder with multiorgan complications, including reproductive system dysfunction where lipid imbalance of germ cells play an important role. N-stearoylethanolamine (NSe) shows a modulatory effect on the lipid composition under different pathologies. Therefore, the aim of our study was to investigate the NSE effect on the testes lipid composition and testosterone level in plasma of diabetic rats. diabetes was induced in Sprague-dawley rats by a single streptozotocin injection (50 mg/kg). animals with glucose levels of 8-12 mmol/l were further selected. NSe was administrated to rats (50 mg/kg) for 10 days at 1.5 months after the streptozotocin injection. The rat testes were used for lipid analysis, namely, phospholipid level, fatty acid methyl esters and plasma testosterone estimation. NSe administration to diabetic rats triggered normalization of total and individual phospholipid content, as well as composition of free and phospholipids fatty acids in the rat testes. In addition, the testosterone content showed a slight increase under the action of NSe. Our results showed that the early stages of diabetes caused destructive changes in rat testes that may induce a decrease in future testicular function. NSe administration to diabetic rats normalized the lipid content of rat testes and was correlated with an increased testosterone level. NSe induced the restoration of testes structure and function during the early stages of streptozotocin-іnduced diabetes in rats.
{"title":"The effect of N-stearoylethanolamine on the lipid composition of the rat testes and testosterone level during the early stages of streptozotocin-іnduced diabetes","authors":"Onopchenko Ov, Horid'ko Tm, Kosiakova Hv","doi":"10.15407/ubj92.02.045","DOIUrl":"https://doi.org/10.15407/ubj92.02.045","url":null,"abstract":"diabetes is a metabolic disorder with multiorgan complications, including reproductive system dysfunction where lipid imbalance of germ cells play an important role. N-stearoylethanolamine (NSe) shows a modulatory effect on the lipid composition under different pathologies. Therefore, the aim of our study was to investigate the NSE effect on the testes lipid composition and testosterone level in plasma of diabetic rats. diabetes was induced in Sprague-dawley rats by a single streptozotocin injection (50 mg/kg). animals with glucose levels of 8-12 mmol/l were further selected. NSe was administrated to rats (50 mg/kg) for 10 days at 1.5 months after the streptozotocin injection. The rat testes were used for lipid analysis, namely, phospholipid level, fatty acid methyl esters and plasma testosterone estimation. NSe administration to diabetic rats triggered normalization of total and individual phospholipid content, as well as composition of free and phospholipids fatty acids in the rat testes. In addition, the testosterone content showed a slight increase under the action of NSe. Our results showed that the early stages of diabetes caused destructive changes in rat testes that may induce a decrease in future testicular function. NSe administration to diabetic rats normalized the lipid content of rat testes and was correlated with an increased testosterone level. NSe induced the restoration of testes structure and function during the early stages of streptozotocin-іnduced diabetes in rats.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"45-58"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43931529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. Nasadyuk, E. A. Sogomonyan, A. Yashchenko, A. Sklyarov
assessment of glycoconjugate expression on cell membranes using the lectin histochemistry technique may be a feasible approach for evaluating the functional state of the cell. The aim of this study was to evaluate carbohydrate determinants of rat stomach mucosa cell membranes under the conditions of CoX-1/2 blockage with indomethacin and pretreatment with the tripeptide h-glu-asp-gly-oh. Male Wistar rats were divided into 3 groups (n = 6 per group): 1st group (control) received vehicle; 2nd – indomethacin (35 mg/kg); 3rd – H-Glu-Asp-Gly-OH (10 μg) 30 min before indomethacin. Rats were sacrificed 24 hours later. gastric mucosa (gM) carbohydrate determinants were studied by lectin-peroxidase technique. The lectins panel included α-fucose(LABA), syalo(WGA, SNA), mannose(Con A, LCA) and galactose-specific (HPA, PNA, SBA) lectins. Intensity of lectin-receptor reaction was scored: 0 – no reaction; 1 – weak; 2 – mild; and 3 – strong reaction. CoX-1/2 blockage caused gM lesions, attenuated by h-glu-asp-gly-oh. Wga and SNa showed the highest affinity to GM. Indomethacin decreased SNA-labeling of epitheliocytes and mucocytes and laBa-labeling of chief cells. h-glu-asp-gly-oh reversed the glycosylation changes, caused by CoX1/CoX-2 blockage only in regards to labeling of chief cells with laBa, epitheliocytes and mucocytes with LCA, mucocytes with SNA. Predominantly H-Glu-Asp-Gly-OH under COX-1/COX-2 blockage had an effect opposite to indomethacin alone but glycosylation changes under these conditions differed significantly also from the control. CoX-1/CoX-2 blockage causes alteration of glycosylation processes in rat gM, mainly reduction of NeuNAc(α2-6)DGal and α-Fuc content. H-Glu-Asp-Gly-OH under the conditions of COX-1/COX-2 blockage leads to more profound changes in GM lectin-binding pattern compared to the independent effect of indomethacin and to control.
{"title":"Lectinocytochemical study of rat stomach mucosa under the conditions of cyclooxygenase-1/-2 blockage and pretreatment witH H-Glu-Asp-Gly-OH","authors":"C. Nasadyuk, E. A. Sogomonyan, A. Yashchenko, A. Sklyarov","doi":"10.15407/ubj92.02.033","DOIUrl":"https://doi.org/10.15407/ubj92.02.033","url":null,"abstract":"assessment of glycoconjugate expression on cell membranes using the lectin histochemistry technique may be a feasible approach for evaluating the functional state of the cell. The aim of this study was to evaluate carbohydrate determinants of rat stomach mucosa cell membranes under the conditions of CoX-1/2 blockage with indomethacin and pretreatment with the tripeptide h-glu-asp-gly-oh. Male Wistar rats were divided into 3 groups (n = 6 per group): 1st group (control) received vehicle; 2nd – indomethacin (35 mg/kg); 3rd – H-Glu-Asp-Gly-OH (10 μg) 30 min before indomethacin. Rats were sacrificed 24 hours later. gastric mucosa (gM) carbohydrate determinants were studied by lectin-peroxidase technique. The lectins panel included α-fucose(LABA), syalo(WGA, SNA), mannose(Con A, LCA) and galactose-specific (HPA, PNA, SBA) lectins. Intensity of lectin-receptor reaction was scored: 0 – no reaction; 1 – weak; 2 – mild; and 3 – strong reaction. CoX-1/2 blockage caused gM lesions, attenuated by h-glu-asp-gly-oh. Wga and SNa showed the highest affinity to GM. Indomethacin decreased SNA-labeling of epitheliocytes and mucocytes and laBa-labeling of chief cells. h-glu-asp-gly-oh reversed the glycosylation changes, caused by CoX1/CoX-2 blockage only in regards to labeling of chief cells with laBa, epitheliocytes and mucocytes with LCA, mucocytes with SNA. Predominantly H-Glu-Asp-Gly-OH under COX-1/COX-2 blockage had an effect opposite to indomethacin alone but glycosylation changes under these conditions differed significantly also from the control. CoX-1/CoX-2 blockage causes alteration of glycosylation processes in rat gM, mainly reduction of NeuNAc(α2-6)DGal and α-Fuc content. H-Glu-Asp-Gly-OH under the conditions of COX-1/COX-2 blockage leads to more profound changes in GM lectin-binding pattern compared to the independent effect of indomethacin and to control.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"33-43"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46818211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
O. Tkachenko, Ie. A. Hudz, Kosiakova Hv, P. Klymenko
In this work we aimed to test the atherosclerotic changes in the aortic wall and pro-coagulant response of the blood coagulation system of spontaneously hypertensive rats (Shr) fed cholesterol-rich diet (CRD) and to study the effect of the anti-inflammatory agent N-stearoylethanolamine (NSE) on the development of atherosclerosis in this model. Female rats (n = 30) with genetically determined hypertension proven by direct measurement of blood pressure were fed crd (5% cholesterol) for 2 months. control group of Shr (n = 10) received standard pellet diet, 10 were fed crd and 10 received crd with daily per os application of NSe at a dose of 50 mg/kg of body weight. histological analysis detected swelling and detachment of endothelial cells, huge edema of the subendothelial layer and a disruption of the middle shell integrity. crd rats had higher fibrinogen concentration, increased rate of platelet aggregation and decreased level of anticoagulant Pc. Platelet aggregation speed increased in crd-fed rats (52.5±4.1%/min) was slightly normalized under the action of NSe (40±8.3 vs 35±9%/min in controls). Fibrinogen concentration was slightly increased in crdfed rats (2.75±0.7 vs 1.9±0.5 mg/ml in controls). however, the level of anticoagulant Pc that was decreased in crd-fed rats (65±16 vs 100±11% in controls) was normalized under the action of NSe (92±17%). NSe also influenced the aorta architecture, however normalizing the thickness of the aorticwall did not change the cholesterol-induced inclusions within aorta media. NSE anti-inflammatory action changes the atherogenic processes in CRD-fed rats mainly protecting PC from consumption during the inflammatory process and reducing edema of the aorta. however hematological parameters (including clotting time in the aPTT test and fibrinogen concentration) changed independently on NSE application. Anti-aggregatory action of NSE on platelets can be a result of direct action on platelets or the consequence of its anti-inflammatory action. During atherogenesis induced by CRD in the model, NSE demonstrated valuable anti-inflammatory action protecting the organism during atherogenesis, however it cannot be assumed as an antithrombotic or antiatherogenic agent because it is unable to influence hemostasis directly.
{"title":"Protective action of N-stearoylethanolamine on blood coagulation and arterial changes in spontaneously hypertensive rats fed cholesterol-rich diet","authors":"O. Tkachenko, Ie. A. Hudz, Kosiakova Hv, P. Klymenko","doi":"10.15407/ubj92.02.060","DOIUrl":"https://doi.org/10.15407/ubj92.02.060","url":null,"abstract":"In this work we aimed to test the atherosclerotic changes in the aortic wall and pro-coagulant response of the blood coagulation system of spontaneously hypertensive rats (Shr) fed cholesterol-rich diet (CRD) and to study the effect of the anti-inflammatory agent N-stearoylethanolamine (NSE) on the development of atherosclerosis in this model. Female rats (n = 30) with genetically determined hypertension proven by direct measurement of blood pressure were fed crd (5% cholesterol) for 2 months. control group of Shr (n = 10) received standard pellet diet, 10 were fed crd and 10 received crd with daily per os application of NSe at a dose of 50 mg/kg of body weight. histological analysis detected swelling and detachment of endothelial cells, huge edema of the subendothelial layer and a disruption of the middle shell integrity. crd rats had higher fibrinogen concentration, increased rate of platelet aggregation and decreased level of anticoagulant Pc. Platelet aggregation speed increased in crd-fed rats (52.5±4.1%/min) was slightly normalized under the action of NSe (40±8.3 vs 35±9%/min in controls). Fibrinogen concentration was slightly increased in crdfed rats (2.75±0.7 vs 1.9±0.5 mg/ml in controls). however, the level of anticoagulant Pc that was decreased in crd-fed rats (65±16 vs 100±11% in controls) was normalized under the action of NSe (92±17%). NSe also influenced the aorta architecture, however normalizing the thickness of the aorticwall did not change the cholesterol-induced inclusions within aorta media. NSE anti-inflammatory action changes the atherogenic processes in CRD-fed rats mainly protecting PC from consumption during the inflammatory process and reducing edema of the aorta. however hematological parameters (including clotting time in the aPTT test and fibrinogen concentration) changed independently on NSE application. Anti-aggregatory action of NSE on platelets can be a result of direct action on platelets or the consequence of its anti-inflammatory action. During atherogenesis induced by CRD in the model, NSE demonstrated valuable anti-inflammatory action protecting the organism during atherogenesis, however it cannot be assumed as an antithrombotic or antiatherogenic agent because it is unable to influence hemostasis directly.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"60-70"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45510522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Thiazole derivatives have cytotoxic effects towards tumor cells, such as glioblastoma, melanoma, leukemia and lymphoma. However, the intracellular mechanism of this action is not clear. The aim of our study was to investigate the action of N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) and 7-benzyl-8-methyl-2-propylpyrazolo[4,3-e]thiazolo[3,2-a]pyrimidin-4(2H)-one (PP2) on cellular structure, and bioenergetic functions of mitochondria in Nemeth-Kellner lymphoma cells (NK/Ly). The structure of treated NK/Ly cells and their mitochondria was examined using electron microscopy. The rate of oxygen uptake by isolated mitochondria was recorded by a polarographic method using a Clark electrode. The mitochondrial potential relative values were registered using fluorescence dye rhodamine 123. In the short-term (15 min), incubation with BF1 and PP2 in 10 and 50 μM concentrations induced apoptotic and necrotic changes in the structure of NK/Ly cells, such as fragmentation and disintegration of the nucleus, destruction of the plasma membrane, and an increase in numbers of lysosomes and mitochondria. a polarographic method did not show significant metabolic shifts in lymphoma mitochondria, in either in vitro or ex vivo actions of the thiazole derivatives. However, fluorescent microscopy showed a significant decrease in mitochondria potential, following a 15 min incubation of cells with 50 μM of PP2. Thus, the electron and fluorescent microscopy data suggest that mitochondria are involved in the mechanism of cytotoxic action of the studied thiazole derivatives.
{"title":"Effects of thiazole derivatives on intracellular structure and functions in murine lymphoma cells","authors":"V. Hreniukh, N. Finiuk, Ya. R. Shalai","doi":"10.15407/ubj92.02.121","DOIUrl":"https://doi.org/10.15407/ubj92.02.121","url":null,"abstract":"Thiazole derivatives have cytotoxic effects towards tumor cells, such as glioblastoma, melanoma, leukemia and lymphoma. However, the intracellular mechanism of this action is not clear. The aim of our study was to investigate the action of N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) and 7-benzyl-8-methyl-2-propylpyrazolo[4,3-e]thiazolo[3,2-a]pyrimidin-4(2H)-one (PP2) on cellular structure, and bioenergetic functions of mitochondria in Nemeth-Kellner lymphoma cells (NK/Ly). The structure of treated NK/Ly cells and their mitochondria was examined using electron microscopy. The rate of oxygen uptake by isolated mitochondria was recorded by a polarographic method using a Clark electrode. The mitochondrial potential relative values were registered using fluorescence dye rhodamine 123. In the short-term (15 min), incubation with BF1 and PP2 in 10 and 50 μM concentrations induced apoptotic and necrotic changes in the structure of NK/Ly cells, such as fragmentation and disintegration of the nucleus, destruction of the plasma membrane, and an increase in numbers of lysosomes and mitochondria. a polarographic method did not show significant metabolic shifts in lymphoma mitochondria, in either in vitro or ex vivo actions of the thiazole derivatives. However, fluorescent microscopy showed a significant decrease in mitochondria potential, following a 15 min incubation of cells with 50 μM of PP2. Thus, the electron and fluorescent microscopy data suggest that mitochondria are involved in the mechanism of cytotoxic action of the studied thiazole derivatives.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"121-130"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44418409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Anna Kryshchyshyn, D. Kaminskyy, O. Roman, R. Kralovics, O. Karpenko, R. Lesyk
A series of novel 2-(5-ylidene-4-oxo-2-thioxo-thiazolidin-3-yl)-succinimides and 5-ylidene-3-(1-arylpyrrolidine-2,5-dione)-thiazolidine-2,4-diones were synthesized. An efficient simple protocol for rhodaninepyrrolidinedione hybrids synthesis which allows avoiding the step of anhydride formation was proposed. Following the previous data on antileukemic properties of related thiazolidinone derivatives, the activity of 19 target compounds was investigated towards four leukemia cell lines: Dami, hl-60, Jurkat, and K562. Among the tested compounds, 3-[5-(4-chloro-benzylidene)-4-oxo-2-thioxo-thiazolidin-3-yl]-1-phenyl-pyrrolidine-2,5-dione (compound 1) possessed good and selective antiproliferative action against Dami and hl-60 cell lines and satisfactory toxicity level (acute toxicity evaluated in vivo in mice).
{"title":"Synthesis and anti-leukemic activity of pyrrolidinedione-thiazolidinone hybrids","authors":"Anna Kryshchyshyn, D. Kaminskyy, O. Roman, R. Kralovics, O. Karpenko, R. Lesyk","doi":"10.15407/ubj92.02.108","DOIUrl":"https://doi.org/10.15407/ubj92.02.108","url":null,"abstract":"A series of novel 2-(5-ylidene-4-oxo-2-thioxo-thiazolidin-3-yl)-succinimides and 5-ylidene-3-(1-arylpyrrolidine-2,5-dione)-thiazolidine-2,4-diones were synthesized. An efficient simple protocol for rhodaninepyrrolidinedione hybrids synthesis which allows avoiding the step of anhydride formation was proposed. Following the previous data on antileukemic properties of related thiazolidinone derivatives, the activity of 19 target compounds was investigated towards four leukemia cell lines: Dami, hl-60, Jurkat, and K562. Among the tested compounds, 3-[5-(4-chloro-benzylidene)-4-oxo-2-thioxo-thiazolidin-3-yl]-1-phenyl-pyrrolidine-2,5-dione (compound 1) possessed good and selective antiproliferative action against Dami and hl-60 cell lines and satisfactory toxicity level (acute toxicity evaluated in vivo in mice).","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"108-119"},"PeriodicalIF":0.0,"publicationDate":"2020-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47389619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Polokhina, D. Kytova, A. Shtemenko, N. Shtemenko
Earlier we have shown that cluster rhenium compounds not only inhibited tumor growth in vivo but also supported the antioxidant state of experimental animals. Further investigation of new dirhenium(III) and cluster rhenium compounds in human leukemic cells is of great importance. the aim of the recent work was to investigate the cytotoxic activity of the new cluster rhenium compound with β-alanine ligands [Re2Cl6(C3h7No2)2]·1.5h2o (I) in the solutions and nanoliposomes alone and together with cisplatin in Jurkat cells. It was shown that I in solution had cytotoxicity close to cisplatin (lC50 = 2.06·10 -6 m). the administration of the rhenium-platinum system with І showed increased cytotoxic activity, especially high when both components of the system were in the mixed liposomes together (lC50 = 4.93·10 -10 m). the new dirhenium dicarboxylate complex with zwitterionic amino acid ligands possesses an appreciable cytotoxic and proapoptotic activity against leukemic cells, especially in combination with cisplatin, guiding the search for novel active rhenium compounds and development of improved regimens for combined chemotherapy based on combination of rhenium-platinum compounds.
{"title":"Cytotoxic activity of the cluster rhenium compound with ?-alanine ligands","authors":"K. Polokhina, D. Kytova, A. Shtemenko, N. Shtemenko","doi":"10.15407/ubj92.01.120","DOIUrl":"https://doi.org/10.15407/ubj92.01.120","url":null,"abstract":"Earlier we have shown that cluster rhenium compounds not only inhibited tumor growth in vivo but also supported the antioxidant state of experimental animals. Further investigation of new dirhenium(III) and cluster rhenium compounds in human leukemic cells is of great importance. the aim of the recent work was to investigate the cytotoxic activity of the new cluster rhenium compound with β-alanine ligands [Re2Cl6(C3h7No2)2]·1.5h2o (I) in the solutions and nanoliposomes alone and together with cisplatin in Jurkat cells. It was shown that I in solution had cytotoxicity close to cisplatin (lC50 = 2.06·10 -6 m). the administration of the rhenium-platinum system with І showed increased cytotoxic activity, especially high when both components of the system were in the mixed liposomes together (lC50 = 4.93·10 -10 m). the new dirhenium dicarboxylate complex with zwitterionic amino acid ligands possesses an appreciable cytotoxic and proapoptotic activity against leukemic cells, especially in combination with cisplatin, guiding the search for novel active rhenium compounds and development of improved regimens for combined chemotherapy based on combination of rhenium-platinum compounds.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"120-126"},"PeriodicalIF":0.0,"publicationDate":"2020-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45322670","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. Gerashchenko, I. Vagina, Y. V. Vagin, V. Kashuba
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 CAFand 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.
{"title":"Pattern of expression of immune- and stroma-associated genes in blood of mice with experimental B16 melanoma","authors":"G. Gerashchenko, I. Vagina, Y. V. Vagin, V. Kashuba","doi":"10.15407/ubj92.01.005","DOIUrl":"https://doi.org/10.15407/ubj92.01.005","url":null,"abstract":"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 CAFand 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.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"5-11"},"PeriodicalIF":0.0,"publicationDate":"2020-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48773814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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.
{"title":"Profiling of metabolic biomarkers in the serum of prostate cancer patients","authors":"F. Ali, S. Akram, S. Niaz, N. Wajid","doi":"10.15407/ubj92.01.056","DOIUrl":"https://doi.org/10.15407/ubj92.01.056","url":null,"abstract":"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.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"56-65"},"PeriodicalIF":0.0,"publicationDate":"2020-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41419462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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 gelfiltration 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.
{"title":"Plasminogen modulates formation and release of platelet angiogenic regulators","authors":"A. Tykhomyrov, D. Zhernosekov, T. Grinenko","doi":"10.15407/ubj92.01.031","DOIUrl":"https://doi.org/10.15407/ubj92.01.031","url":null,"abstract":"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 gelfiltration 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.","PeriodicalId":23448,"journal":{"name":"Ukrainian Biochemical Journal","volume":"92 1","pages":"31-40"},"PeriodicalIF":0.0,"publicationDate":"2020-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48502917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：