Vaccination is the best method to prevent the spread of infectious diseases, its disadvantages are side effects. Potentially safe DNA, RNA or protein molecules possess antigenic properties, but are low-immunogenic and therefore require conjugation with an adjuvant. The aim of the research was to evaluate Chitosan (CS) potency as an adjuvant and compare its effectiveness depending on the route of drug administration. The experiments were carried out on 3 groups of BALB/c mice. Mice of the first group were injected subcutaneously with 20 µl of a mixture of CS (3.3 mg/kg) and BSA (1.7 mg/kg). The mixture of CS and BSA at the same doses and volume was administered orally to mice of the second experimental group. The third group – control – unvaccinated mice. Anti-BSA antibody levels were measured by ELISA. Aspartate aminotransferase, alanine aminotransferase activity and cholesterol, creatinine and urea levels were determined in the serum. It was found that both subcutaneous and mucosal immunizations provided a 2-fold increase in anti-BSA antibody titers against the background of maintaining all biochemical blood parameters at the level of the physiological norm. However, AST activity in the serum of oral-immunized mice was elevated as compared to subcutaneous-immunized mice. Serum cholesterol level in the group of subcutaneously immunized mice and creatinine and urea levels in both experimental groups were reduced compared to the control. It is concluded that oral immunization with CS is the optimal route for antigen-specific IgG antibody response induction.
{"title":"Comparison of adjuvant properties of chitosan during oral and subcutaneous immunization of mice with BSA","authors":"M. Kozak, I. Petruh, V. Vlizlo","doi":"10.15407/ubj94.02.031","DOIUrl":"https://doi.org/10.15407/ubj94.02.031","url":null,"abstract":"Vaccination is the best method to prevent the spread of infectious diseases, its disadvantages are side effects. Potentially safe DNA, RNA or protein molecules possess antigenic properties, but are low-immunogenic and therefore require conjugation with an adjuvant. The aim of the research was to evaluate Chitosan (CS) potency as an adjuvant and compare its effectiveness depending on the route of drug administration. The experiments were carried out on 3 groups of BALB/c mice. Mice of the first group were injected subcutaneously with 20 µl of a mixture of CS (3.3 mg/kg) and BSA (1.7 mg/kg). The mixture of CS and BSA at the same doses and volume was administered orally to mice of the second experimental group. The third group – control – unvaccinated mice. Anti-BSA antibody levels were measured by ELISA. Aspartate aminotransferase, alanine aminotransferase activity and cholesterol, creatinine and urea levels were determined in the serum. It was found that both subcutaneous and mucosal immunizations provided a 2-fold increase in anti-BSA antibody titers against the background of maintaining all biochemical blood parameters at the level of the physiological norm. However, AST activity in the serum of oral-immunized mice was elevated as compared to subcutaneous-immunized mice. Serum cholesterol level in the group of subcutaneously immunized mice and creatinine and urea levels in both experimental groups were reduced compared to the control. It is concluded that oral immunization with CS is the optimal route for antigen-specific IgG antibody response induction.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"27 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73331967","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}
V. Stetska, T. Dovbynchuk, N. Dziubenko, A. Zholos, G. Tolstanova
Parkinson’s disease (PD) is neurodegenerative disease, which is accompanied by degeneration of dopaminergic neurons in subtantia nigra. Non-motor symptoms, in particular, disorders of the gastrointestinal (GI) tract are observed in 20-80% of patients some 15-20 years before clinically diagnosed PD and are not a least important feature of PD pathogenesis. The transient receptor potential (TRP) channels are expressed throughout the GI tract, where they play an important role in taste, thermoregulation, pain, mucosal function and homeostasis, control of interstitial motility etc. The aim of this study was to investigate the contribution of TRPV4 and TRPM8 channels in the GI motor function in the colon of rats with PD, incduced by injection of the 12 μg 6-hydroxydopamine (6-OHDA). The studies were performed on the 4th week and the 7th month after PD induction The rats were randomly divided into: I group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 4 weeks after injection (n = 5); II group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 4 weeks after injection (n = 5); III group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 7 months after injection (n = 4); IV group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 7 months after injection (n = 5). We evaluated the body weight of rats, GI transit time, the cecum weight index and immunohistochemical identification of tyrosine hydroxylase (TH) -positive cells, and TRPV4, TRPM8 expression in rat’s colon. We showed that on the 7th month of the experiment, the GI transit time doubles over time; the cecum weight index of 6-OHDA rats increased by 57%; the number of TH-positive cells in colon rats decreased 2-fold, while TRPM8 ion channels were downregulated in PD rats and TRPV4 ion channels were upregulated in the colon of rats with 6-OHDA-PD. It was concluded that TRPV4 and TRPM8 ion channels may be considered pharmacological targets in the progression of PD pathology.
{"title":"Changes in the expression of TRPV4 and TRPM8 channels in the colon of rats with 6-OHDA-induced Parkinson’s disease","authors":"V. Stetska, T. Dovbynchuk, N. Dziubenko, A. Zholos, G. Tolstanova","doi":"10.15407/ubj94.02.057","DOIUrl":"https://doi.org/10.15407/ubj94.02.057","url":null,"abstract":"Parkinson’s disease (PD) is neurodegenerative disease, which is accompanied by degeneration of dopaminergic neurons in subtantia nigra. Non-motor symptoms, in particular, disorders of the gastrointestinal (GI) tract are observed in 20-80% of patients some 15-20 years before clinically diagnosed PD and are not a least important feature of PD pathogenesis. The transient receptor potential (TRP) channels are expressed throughout the GI tract, where they play an important role in taste, thermoregulation, pain, mucosal function and homeostasis, control of interstitial motility etc. The aim of this study was to investigate the contribution of TRPV4 and TRPM8 channels in the GI motor function in the colon of rats with PD, incduced by injection of the 12 μg 6-hydroxydopamine (6-OHDA). The studies were performed on the 4th week and the 7th month after PD induction The rats were randomly divided into: I group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 4 weeks after injection (n = 5); II group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 4 weeks after injection (n = 5); III group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 7 months after injection (n = 4); IV group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 7 months after injection (n = 5). We evaluated the body weight of rats, GI transit time, the cecum weight index and immunohistochemical identification of tyrosine hydroxylase (TH) -positive cells, and TRPV4, TRPM8 expression in rat’s colon. We showed that on the 7th month of the experiment, the GI transit time doubles over time; the cecum weight index of 6-OHDA rats increased by 57%; the number of TH-positive cells in colon rats decreased 2-fold, while TRPM8 ion channels were downregulated in PD rats and TRPV4 ion channels were upregulated in the colon of rats with 6-OHDA-PD. It was concluded that TRPV4 and TRPM8 ion channels may be considered pharmacological targets in the progression of PD pathology.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87071270","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}
I. Mykhaloiko, R. Yatsyshyn, N. Cherniuk, M. Humeniuk
The aim of research was to study the levels of thrombomodulin (TM) and von Willebrand factor (VWF) in the serum and urine of patients with chronic kidney disease (CKD)as diagnostic markers of endothelial dysfunction. The study involved 140 patients with CKD. The clinical diagnosis was determined based on standard methods of patients examination according to the kidney diseases classification and protocols of CKD patients management. The concentrations of TM and VWF in serum and urine were quantified by ELISA. A generalized endothelial dysfunction in the vessels of the whole body, including the kidneys and high concentration of TM and FVF in the serum and urine of patients with a diabetic nephropathy have been found. The concentration of TM and VWF in the serum of patients with a chronic glomerulonephritis was at the same level as in the serum of healthy individuals, while those in urine significantly exceeded the control values, indicating endothelial damage in the glomeruli of the kidneys due to exposure to pro-inflammatory cytokines. In our opinion, the studied markers will contribute to the timely diagnosis of endothelial dysfunction in patients with CKD and to the development of criteria for prescribing antiplatelet agents in glomerular kidney disease.
{"title":"Thrombomodulin and von willebrand factor as markers of endothelial dysfunction in patients with chronic kidney disease","authors":"I. Mykhaloiko, R. Yatsyshyn, N. Cherniuk, M. Humeniuk","doi":"10.15407/ubj94.02.045","DOIUrl":"https://doi.org/10.15407/ubj94.02.045","url":null,"abstract":"The aim of research was to study the levels of thrombomodulin (TM) and von Willebrand factor (VWF) in the serum and urine of patients with chronic kidney disease (CKD)as diagnostic markers of endothelial dysfunction. The study involved 140 patients with CKD. The clinical diagnosis was determined based on standard methods of patients examination according to the kidney diseases classification and protocols of CKD patients management. The concentrations of TM and VWF in serum and urine were quantified by ELISA. A generalized endothelial dysfunction in the vessels of the whole body, including the kidneys and high concentration of TM and FVF in the serum and urine of patients with a diabetic nephropathy have been found. The concentration of TM and VWF in the serum of patients with a chronic glomerulonephritis was at the same level as in the serum of healthy individuals, while those in urine significantly exceeded the control values, indicating endothelial damage in the glomeruli of the kidneys due to exposure to pro-inflammatory cytokines. In our opinion, the studied markers will contribute to the timely diagnosis of endothelial dysfunction in patients with CKD and to the development of criteria for prescribing antiplatelet agents in glomerular kidney disease.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"84 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76558900","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}
The review article presents the author’s model of one of the blocks of the integrated adaptation mechanism to physical activity and the accompanying moderate heat effects. The participation of heat shock proteins in the stabilization of the tertiary structure and in the restoration of the function of proteins damaged by temperature and physical stressors but performing catalytic, transport, reception or protective role and being involved in the processes of contraction- relaxation and muscle and bone tissue remodeling is discussed.
{"title":"Heat shock proteins in adaptation to physical activity","authors":"V. Kuibida, P. Kokhanets, V. Lopatynska","doi":"10.15407/ubj94.02.005","DOIUrl":"https://doi.org/10.15407/ubj94.02.005","url":null,"abstract":"The review article presents the author’s model of one of the blocks of the integrated adaptation mechanism to physical activity and the accompanying moderate heat effects. The participation of heat shock proteins in the stabilization of the tertiary structure and in the restoration of the function of proteins damaged by temperature and physical stressors but performing catalytic, transport, reception or protective role and being involved in the processes of contraction- relaxation and muscle and bone tissue remodeling is discussed.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77218563","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}
The development of methods for the detection of polyamines in biological fluids is essential to improve early diagnosis and treatment of patients with prostate cancer. One of the promising areas is the use of noble metal nanoparticles. According to the literature data, there is no methodological approach have been developed to reliably distinguish spermine from other polyamines, in particular, from their acetylated forms and related compounds present in biological fluids. The paper presents the results of spectrophotometric determination of spermine both alone and in the presence of putrescine, spermidine or urea in the urine using gold nanoparticles. The results of the experiments proved that the developed method is suitable for the selective determination of spermine. It was shown that the presence of spermidine, putrescine, acetylated forms of polyamines or carbamide does not affect the results of the analysis.
{"title":"Application of gold nanoparticles to determine spermine in the presence of other polyamines","authors":"Y. Yanish, M. Prylutskyi, S. Zaletok, Y. Mukha","doi":"10.15407/ubj94.02.066","DOIUrl":"https://doi.org/10.15407/ubj94.02.066","url":null,"abstract":"The development of methods for the detection of polyamines in biological fluids is essential to improve early diagnosis and treatment of patients with prostate cancer. One of the promising areas is the use of noble metal nanoparticles. According to the literature data, there is no methodological approach have been developed to reliably distinguish spermine from other polyamines, in particular, from their acetylated forms and related compounds present in biological fluids. The paper presents the results of spectrophotometric determination of spermine both alone and in the presence of putrescine, spermidine or urea in the urine using gold nanoparticles. The results of the experiments proved that the developed method is suitable for the selective determination of spermine. It was shown that the presence of spermidine, putrescine, acetylated forms of polyamines or carbamide does not affect the results of the analysis.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"12 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85050385","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}
One of the most important pathogenetic mechanisms of autoimmune thyroiditis (AIT) is the violation of immunological tolerance and the development of the autoimmune process, the markers of which are various biologically active substances, in particular, matrix metalloproteinases (MMP) of the extracellular matrix (ECM). MMPs play a crucial role in the development of pathological processes in these diseases, contributing to matrix degradation due to imbalance between the activity of enzymes and their inhibitors. The aim of the work was to study the activity of key metalloproteinases and the level of α2-macroglobulin in patients with autoimmune thyroiditis. The diagnosis of AIT was established based on the study of data on anamnesis, thyroid status, the results of ultrasound of TG, and the presence of antibodies to the thyroid-stimulating hormone receptor (TSH) in blood plasma. Patients were enrolled in 2 groups: group 1 – 74 patients with a manifest form of the disease; group 2 – 96 patients with a subclinical form of the disease. The study of matrix metalloprotein activity in the examined patients showed a statistically significant (P = 0.015) increase in MMP-3 and MMP-7 activity in patients with AIT compared to the corresponding parameters in persons of the control group. Thus, levels of MMP-3 and 7 were in the group of patients, respectively 56 (51.0; 59.0) and 4.6 (4.3; 5.2) ng/ml, in control 23.0 (16.0; 26.0) and 3.6 (3.4; 4.1) ng/ml, respectively.
{"title":"Study of matrix metalloproteinase activity in patients with autoimmune thyroiditis","authors":"R. Rahimova","doi":"10.15407/ubj94.02.051","DOIUrl":"https://doi.org/10.15407/ubj94.02.051","url":null,"abstract":"One of the most important pathogenetic mechanisms of autoimmune thyroiditis (AIT) is the violation of immunological tolerance and the development of the autoimmune process, the markers of which are various biologically active substances, in particular, matrix metalloproteinases (MMP) of the extracellular matrix (ECM). MMPs play a crucial role in the development of pathological processes in these diseases, contributing to matrix degradation due to imbalance between the activity of enzymes and their inhibitors. The aim of the work was to study the activity of key metalloproteinases and the level of α2-macroglobulin in patients with autoimmune thyroiditis. The diagnosis of AIT was established based on the study of data on anamnesis, thyroid status, the results of ultrasound of TG, and the presence of antibodies to the thyroid-stimulating hormone receptor (TSH) in blood plasma. Patients were enrolled in 2 groups: group 1 – 74 patients with a manifest form of the disease; group 2 – 96 patients with a subclinical form of the disease. The study of matrix metalloprotein activity in the examined patients showed a statistically significant (P = 0.015) increase in MMP-3 and MMP-7 activity in patients with AIT compared to the corresponding parameters in persons of the control group. Thus, levels of MMP-3 and 7 were in the group of patients, respectively 56 (51.0; 59.0) and 4.6 (4.3; 5.2) ng/ml, in control 23.0 (16.0; 26.0) and 3.6 (3.4; 4.1) ng/ml, respectively.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77836295","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}
A. Ojo, A. I. Oni, D. Rotimi, M. Iyobhebhe, P. O. Adeniji, J. Talabi, O. A. Ojo
The prevalence of diabetes, as reported by the World Health Organization and the International Diabetes Federation, has raised many eyebrows about the dangers of diabetes mellitus to society, leading to the development of various therapeutic techniques, including nanotechnological, in the management of this disease. This review discusses silver, gold, ceramic, alloy, magnetic, silica, polymeric nanoparticles and their various applications in diabetes management which may help to reduce the incidence of diabetes and its complication.
{"title":"Nanoparticles application as a therapeutic strategy for diabetes mellitus management","authors":"A. Ojo, A. I. Oni, D. Rotimi, M. Iyobhebhe, P. O. Adeniji, J. Talabi, O. A. Ojo","doi":"10.15407/ubj94.02.015","DOIUrl":"https://doi.org/10.15407/ubj94.02.015","url":null,"abstract":"The prevalence of diabetes, as reported by the World Health Organization and the International Diabetes Federation, has raised many eyebrows about the dangers of diabetes mellitus to society, leading to the development of various therapeutic techniques, including nanotechnological, in the management of this disease. This review discusses silver, gold, ceramic, alloy, magnetic, silica, polymeric nanoparticles and their various applications in diabetes management which may help to reduce the incidence of diabetes and its complication.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"64 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73722578","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}
In the 21st century, none of the scientists denies the determining role of the cardiovascular system and its central organ, the heart. The ongoing attempts to design new medications, elaborate effective trainings, heart transplant programs testify that humanity does not abandon attempts to improve and prolong human life, especially given the fact that the world’s biggest killer is ischemic heart disease. The most significant achievements in this field receive the highest rating in the scientific community – the Nobel Prize. In 1998, the Nobel Prize in Physiology or Medicine was awarded jointly to Robert F. Furchgott, Louis J. Ignarro and Ferid Murad “for their discoveries concerning nitric oxide as a signaling molecule in cardiovascular system”. Their discovery triggered an international boom in research on nitric oxide. The paper aims to outline briefly the main stages of the scientific activity of R.F. Furchgott, L.J. Ignarro and F. Murad.
在21世纪，没有一个科学家否认心血管系统及其中心器官——心脏的决定性作用。不断尝试设计新的药物，精心设计有效的训练，心脏移植项目，证明人类并没有放弃改善和延长人类生命的尝试，特别是考虑到世界上最大的杀手是缺血性心脏病。这一领域最重要的成就获得了科学界的最高评价——诺贝尔奖。1998年，诺贝尔生理学或医学奖联合授予Robert F. Furchgott、Louis J. Ignarro和Ferid Murad，以表彰他们“发现一氧化氮作为心血管系统的信号分子”。他们的发现引发了国际上对一氧化氮研究的热潮。本文旨在简要概述R.F. Furchgott, L.J. Ignarro和F. Murad的科学活动的主要阶段。
{"title":"Unraveling the mystery of nitric oxide: Nobel prize winners Robert Furchgott, Louis Ignarro, and Ferid Murad","authors":"T. Danylova, S. Komisarenko","doi":"10.15407/ubj94.02.085","DOIUrl":"https://doi.org/10.15407/ubj94.02.085","url":null,"abstract":"In the 21st century, none of the scientists denies the determining role of the cardiovascular system and its central organ, the heart. The ongoing attempts to design new medications, elaborate effective trainings, heart transplant programs testify that humanity does not abandon attempts to improve and prolong human life, especially given the fact that the world’s biggest killer is ischemic heart disease. The most significant achievements in this field receive the highest rating in the scientific community – the Nobel Prize. In 1998, the Nobel Prize in Physiology or Medicine was awarded jointly to Robert F. Furchgott, Louis J. Ignarro and Ferid Murad “for their discoveries concerning nitric oxide as a signaling molecule in cardiovascular system”. Their discovery triggered an international boom in research on nitric oxide. The paper aims to outline briefly the main stages of the scientific activity of R.F. Furchgott, L.J. Ignarro and F. Murad.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91259411","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}
L. Babich, S. Shlykov, O. Yesypenko, A. O. Bavelska-Somak, A. G. Zahoruiko, I. Horak, L. Drobot, S. Kosterin
According to our earlier data, calix[4]arene chalcone amides modulate Ca ions exchange in the myometrium mitochondria and the level of inner membrane polarization that can potentially affect cell survival. To test this hypothesis, we studied the effect of calix[4]arene with 4 chalcone amide groups on mitochondria membrane polarization and viability of 4T1 mouse breast adenocarcinoma cells, a surrogate model of human triple-negative breast cancer, and on its highly malignant subline overexpressing the adaptor protein Ruk/CIN85. Mitochondria membrane potential was measured by flow cytometry, and cell viability was assessed using Trypan blue dye exclusion. It was shown that mitochondrial membranes of control (Mock) cells had a higher polarization level (67.80 ± 8.82 r.u., n = 5) compared to 4T1 cells with up-regulation of Ruk/CIN85 (RukUp cells) (25.42 ± 2.58 r.u., n = 4). Upon incubation of cells with 1 μM calix[4]arene C-1011, the CCCP-sensitive component of mitochondrial membranes polarization decreased (by almost 50%) in 4T1 Mock cells and did not change in RukUp cells compared with the control. It was demonstrated that 1 μM calix[4]arene C-1011 suppressed the viability of 4T1 Mock cells by 45%, but did not affect RukUp cells considerably. It was suggested that calix[4]arene chalcone amide С-1011 decreased mouse breast adenocarcinoma 4T1 cell viability at least by affecting mitochondrial membrane polarization.The data obtained indicate the prospects of further studies of calix[4]arene chalcone amide as a potential anticancer drug candidate.
根据我们早期的数据，杯[4]芳烃查尔酮酰胺调节肌层线粒体中的钙离子交换和内膜极化水平，这可能会影响细胞存活。为了验证这一假设，我们研究了带有4个查尔酮酰胺基团的杯[4]芳烯对人类三阴性乳腺癌替代模型4T1小鼠乳腺腺癌细胞线粒体膜极化和活力的影响，以及对其高恶性亚群过表达适配蛋白Ruk/CIN85的影响。流式细胞术测定线粒体膜电位，台盼蓝染色法测定细胞活力。结果表明，对照(Mock)细胞的线粒体膜极化水平(67.80±8.82 r.u, n = 5)高于4T1细胞，RukUp细胞的Ruk/CIN85表达上调(25.42±2.58 r.u, n = 4)。1 μM杯[4]烯C-1011孵育后，4T1模拟细胞的cccp敏感成分线粒体膜极化水平下降(近50%)，而RukUp细胞的cccp敏感成分与对照相比没有变化。结果表明，1 μM杯[4]芳烃C-1011对4T1模拟细胞的活性有45%的抑制作用，但对RukUp细胞没有明显影响。研究表明，杯[4]芳烃查尔酮酰胺С-1011至少通过影响线粒体膜极化降低小鼠乳腺腺癌4T1细胞活力。这些数据表明了杯[4]芳烃查尔酮酰胺作为潜在的抗癌候选药物的进一步研究前景。
{"title":"Calix[4]arene chalcone amide C-1011 elicits differential effects on the viability of 4T1 mouse breast adenocarcinoma cells with different levels of adaptor protein Ruk/CIN85 expression","authors":"L. Babich, S. Shlykov, O. Yesypenko, A. O. Bavelska-Somak, A. G. Zahoruiko, I. Horak, L. Drobot, S. Kosterin","doi":"10.15407/ubj94.02.024","DOIUrl":"https://doi.org/10.15407/ubj94.02.024","url":null,"abstract":"According to our earlier data, calix[4]arene chalcone amides modulate Ca ions exchange in the myometrium mitochondria and the level of inner membrane polarization that can potentially affect cell survival. To test this hypothesis, we studied the effect of calix[4]arene with 4 chalcone amide groups on mitochondria membrane polarization and viability of 4T1 mouse breast adenocarcinoma cells, a surrogate model of human triple-negative breast cancer, and on its highly malignant subline overexpressing the adaptor protein Ruk/CIN85. Mitochondria membrane potential was measured by flow cytometry, and cell viability was assessed using Trypan blue dye exclusion. It was shown that mitochondrial membranes of control (Mock) cells had a higher polarization level (67.80 ± 8.82 r.u., n = 5) compared to 4T1 cells with up-regulation of Ruk/CIN85 (RukUp cells) (25.42 ± 2.58 r.u., n = 4). Upon incubation of cells with 1 μM calix[4]arene C-1011, the CCCP-sensitive component of mitochondrial membranes polarization decreased (by almost 50%) in 4T1 Mock cells and did not change in RukUp cells compared with the control. It was demonstrated that 1 μM calix[4]arene C-1011 suppressed the viability of 4T1 Mock cells by 45%, but did not affect RukUp cells considerably. It was suggested that calix[4]arene chalcone amide С-1011 decreased mouse breast adenocarcinoma 4T1 cell viability at least by affecting mitochondrial membrane polarization.The data obtained indicate the prospects of further studies of calix[4]arene chalcone amide as a potential anticancer drug candidate.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"14 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81785488","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}
M. Lootsik, N. Manko, R. Bilyy, M. Lutsyk (Jr.), R. Stoika
Chitosan is biocompatible and biodegradable natural biopolymer widely applied in various fields of biology, medicine, and pharmacy, however, its effects significantly depend on the degree of polymerization (DP) and the degree of deacetylation (DDA) of polymer chains. Evaluation of the chitosan chain diversity by DP requires the use of a highly expensive method of high-performance size exclusion chromatography. The aim of our study was to determine the molecular weight profile of chitosan specimens by the use of electrophoresis in a porosity step gradient polyacrylamide gel and to evaluate the efficacy of this method in monitoring the purification of chitosan fragments and its derivatives. Two types of step gradient porosity gels were used: 1) gels of layers with acrylamide concentration 2.5, 3.5, 5.0, 10.0, 15.0, 20.0 % w/v for native chitosan or its high molecular fragments; 2) gels of layers with acrylamide concentration 2.5, 5.0, 10.0, 15.0, 20.0, 25.0 % w/v for low molecular chitosan fragments. The main amount of molecules from the chitosan pool was localized in the type 1 gel in the region of 550-40 kDa and distributed among three bands, which in different samples differed significantly in percentage. Electrophoresis of chitosan fragments fractionated by gel permeation chromatography provided a clear separation of medium molecular weight fragments (50–400 kDa) in type 1 gel and of low molecular weight fragments (3–40 kDa) in type 2 gel. Thus the method of chitosan electrophoresis in a step-gradient porosity of polyacrylamide gel was developed which permits to characterize the molecular weight profile of chitosan specimens polymer chains and is effective in monitoring the isolation of chitosan fragments by gel penetration chromatography of molecular weights from 3 to 400 kDa.
{"title":"Analysis of chitosan molecular weight profile by electrophoresis in a porosity step gradient polyacrylamide gel","authors":"M. Lootsik, N. Manko, R. Bilyy, M. Lutsyk (Jr.), R. Stoika","doi":"10.15407/ubj94.02.076","DOIUrl":"https://doi.org/10.15407/ubj94.02.076","url":null,"abstract":"Chitosan is biocompatible and biodegradable natural biopolymer widely applied in various fields of biology, medicine, and pharmacy, however, its effects significantly depend on the degree of polymerization (DP) and the degree of deacetylation (DDA) of polymer chains. Evaluation of the chitosan chain diversity by DP requires the use of a highly expensive method of high-performance size exclusion chromatography. The aim of our study was to determine the molecular weight profile of chitosan specimens by the use of electrophoresis in a porosity step gradient polyacrylamide gel and to evaluate the efficacy of this method in monitoring the purification of chitosan fragments and its derivatives. Two types of step gradient porosity gels were used: 1) gels of layers with acrylamide concentration 2.5, 3.5, 5.0, 10.0, 15.0, 20.0 % w/v for native chitosan or its high molecular fragments; 2) gels of layers with acrylamide concentration 2.5, 5.0, 10.0, 15.0, 20.0, 25.0 % w/v for low molecular chitosan fragments. The main amount of molecules from the chitosan pool was localized in the type 1 gel in the region of 550-40 kDa and distributed among three bands, which in different samples differed significantly in percentage. Electrophoresis of chitosan fragments fractionated by gel permeation chromatography provided a clear separation of medium molecular weight fragments (50–400 kDa) in type 1 gel and of low molecular weight fragments (3–40 kDa) in type 2 gel. Thus the method of chitosan electrophoresis in a step-gradient porosity of polyacrylamide gel was developed which permits to characterize the molecular weight profile of chitosan specimens polymer chains and is effective in monitoring the isolation of chitosan fragments by gel penetration chromatography of molecular weights from 3 to 400 kDa.","PeriodicalId":23007,"journal":{"name":"The Ukrainian Biochemical Journal","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90640058","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：