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Papers by Nikolay Kolmakov
Cell and Tissue Biology, 2019
The pharmacokinetics of two substances containing recombinant insulin-like growth factor 1 (IGF1)... more The pharmacokinetics of two substances containing recombinant insulin-like growth factor 1 (IGF1) as an active substance was considered. The first substance (IGF 1) contains the actual recombinant IGF1, and the second (IGF 2) contains IGF1, which is translated from plasmid DNA encoding the IGF-1 gene for this protein. It was established that, with intramuscular administration of IGF 1 , the delay time for its entry into the bloodstream is 1.5-2 h, while that with IGF 2 is 24-25 h. This indicates the presence of various mechanisms of accumulation of these substances in the systemic circulation. The maximum concentration of IGF 1 in the blood was determined 5 h after administration and that of IGF 2 was determined 125 h after administration. The maximum concentrations of these substances are comparable to each other. The concentration of IGF 1 in the blood decreases to its initial value 12 h after its administration, and the concentration of IGF 2 does so after 216 h. The clearance parameters (Cl) and elimination constants (Kel) of the considered substances also had significant differences, which confirms the presence of fast and slow dynamics of the decrease of their maximum concentrations after intramuscular injection. The different dynamics of the accumulation of substances in the blood and their elimination from the bloodstream after administration, as well as the different values of the area parameters under the pharmacokinetic curve (AUC t , AUC ∞) demonstrate that IGF 2 has been in the systemic circulation for a longer time than IGF 1. This is essential for the appearance and severity of pharmacodynamic effects.
Journal of Applied Ichthyology, 2006
Reproductive toxicology (Elmsford, N.Y.), Dec 23, 2017
Earlier we showed that asymmetric methylation of sister chromatids (AMSC) was a specific characte... more Earlier we showed that asymmetric methylation of sister chromatids (AMSC) was a specific characteristic of differentiation potency, and supposed that AMSC could be a useful marker of environmental impact connected with differentiation and/or dedifferentiation. Here we investigated the level of AMSC in chromosomes and the nuclei methylation in mouse preimplantation and postimplantation embryos, in comparison with the undifferentiated cells of mouse embryonal carcinoma cell line F9, and human differentiated HEK293 cells upon BPA influence. We found that exposure of mouse preimplantation embryos to BPA caused a significant decrease in the level of AMSC in chromosomes and the nuclei methylation. The BPA exposure of potentially differentiating F9 cells had no any influence on DNA methylation in nuclei but significantly decreased the number of AMSC. The level of DNA methylation and AMSC in HEK293 cells were not also changed. These data indicate that BPA exerts significant influence on dif...
Medical academic journal, 2018
MemoriAl plate unveiled in memory of staff of the Institute of Experimental Medicine who were vic... more MemoriAl plate unveiled in memory of staff of the Institute of Experimental Medicine who were victims of political repression
Medical academic journal, 2018
This study aimed to develop a method for producing human recombinant insulin-like growth factor (... more This study aimed to develop a method for producing human recombinant insulin-like growth factor (IGF-1) based on a prokaryotic expression system and to characterize the highly purified protein. To achieve the study’s goal, the following methods were conducted: we performed automated chemical synthesis of DNA, constructed the expression plasmid, obtained Escherichia coli cell-producers of human recombinant IGF-1, cultivated the obtained producer cells with the induction of recombinant protein synthesis by isopropyl-β-D-1-thiogalactopyranoside and lactose, and purified human recombinant IGF-1 with affinity and cation exchange chromatography. The recombinant protein IGF-1 forms inclusion bodies during synthesis in Escherichia coli BL21 cells that contain plasmid pET28-IGF-1. Purified recombinant protein was obtained with a purity of 98% using affinity and cation exchange chromatography methods. The protein yield was 6 mg of human recombinant IGF-1 from 1 g of raw biomass. The resulting...
BioMetals, 2018
Among the properties of lactoferrin (LF) are bactericidal, antianemic, immunomodulatory, antitumo... more Among the properties of lactoferrin (LF) are bactericidal, antianemic, immunomodulatory, antitumour, antiphlogistic effects. Previously we demonstrated its capacity to stabilize in vivo HIF-1alpha and HIF-2-alpha, which are redox-sensitive multiaimed transcription factors. Various tissues of animals receiving recombinant human LF (rhLF) responded by expressing the HIF-1-alpha target genes, hence such proteins as erythropoietin (EPO), ceruloplasmin, etc. were synthesized in noticeable amounts. Among organs in which EPO synthesis occurred were brain, heart, spleen, liver, kidneys and lungs. Other researchers showed that EPO can act as a protectant against severe brain injury and status epilepticus in rats. Therefore, we tried rhLF as a protector against the severe neurologic disorders developed in rats, such as the rotenone-induced model of Parkinson's disease and experimental autoimmune encephalomyelitis as a model of multiple sclerosis, and observed its capacity to mitigate the grave symptoms. Moreover, an
Metallomics, 2017
Ceruloplasmin (Cp) is a copper-containing multifunctional oxidase of plasma. New rat Cp features ... more Ceruloplasmin (Cp) is a copper-containing multifunctional oxidase of plasma. New rat Cp features were discovered by X-ray analysis.
BioMetals, 2016
Strongly pronounced argyrosis caused by adding AgCl to the feed of laboratory rats efficiently mi... more Strongly pronounced argyrosis caused by adding AgCl to the feed of laboratory rats efficiently mimics the deficiency of ceruloplasmin (CP) ferroxidase activity. Bringing the concentration of AgCl in the feedstuff of lactating rats to 250 mg % and keeping their progeny (Ag-rats) for 3 months on the same silvercontaining feed provided the serum iron content 1.4 times lower than that in the control group. Besides, the ferroxidase activity of CP dropped to zero. In CP purified from sera of Ag-rats two copper ions were substituted with two silver ions. Using rat models of both posthemorrhagic and hemolytic anemia we showed that the deficiency of CP ferroxidase activity in Ag-rats affects the iron content in serum, though does not prevent the recovery of hemoglobin level accompanied by exhaustion of iron caches in liver and spleen. When apolactoferrin (apo-LF) was administered to Ag-rats suffering from either post-hemorrhagic or hemolytic anemia, both hemoglobin and serum iron were restored more rapidly than in the control animals. In independent experiments Ag-rats were compared with those fed on regular diet and the former displayed a prolonged 3-day stabilization of hypoxia-inducible factors 1 and 2 alpha (HIF-1a and HIF-2a) along with an increased serum concentration of erythropoietin. Introduction to Ag-rats of active CP separately or together with apo-LF reduced that effect to 1 day only. It is concluded that saturation of apo-LF with iron, provided by active CP, can strongly affect its protective capacity. Keywords Iron metabolism Á Ferroxidase Á Ceruloplasmin Á Lactoferrin Á Transferrin Á Erythropoietin Á Post-hemorrhagic anemia Á Hemolytic anemia Á Hypoxia-inducible factor Abbreviations Ag-rats Rats with AgCl-diet Apo-LF Apo-form LF CP Ceruloplasmin EPO Erythropoietin HIF-1a Hypoxia-inducible factor-1 alpha HIF-2a Hypoxia-inducible factor-2 alpha LF Lactoferrin TF Transferrin
Journal of Applied Ichthyology, 2006
Summary Sturgeon oocytes, like those of other vertebrates, are arrested in early meiosis and requ... more Summary Sturgeon oocytes, like those of other vertebrates, are arrested in early meiosis and require a steroid or steroids to trigger meiosis resumption or maturation. The identity of the maturation-inducing steroid in sturgeons has present study was designed to investigate in sterlet the follicular secretion and relative blood changes of free and conjugated 17, 20βP, 20βS and of the androgens T, 11KT, in response to hormonal treatment and to determine the relative potencies of the progestagens, 17, 20β-P, 20βS, P4 and S, in inducing oocyte final ...
Environmental Science & Technology, 2009
Exposure of olfactory epithelium to environmentally relevant concentrations of copper disrupts ol... more Exposure of olfactory epithelium to environmentally relevant concentrations of copper disrupts olfaction in fish. To examine the dynamics of recovery at both functional and morphological levels after acute copper exposure, unilateral exposure of goldfish olfactory epithelia to 100 µM CuSO 4 (10 min) was followed by electro-olfactogram (EOG) recording and scanning electron microscopy. Sensitivity to amino acids (L-arginine and L-serine), generally considered food-related odorants, recovered most rapidly (three days), followed by that to catecholamines (3-O-methoxytyramine), bile acids (taurolithocholic acid) and the steroid pheromone, 17,20-dihydroxy-4-pregnen-3-one 20-sulfate, which took 28 days to reach full recovery. Sensitivity to the postovulatory pheromone prostaglandin F 2R had not fully recovered even at 28 days. These changes in sensitivity were correlated with changes in the recovery of ciliated and microvillous receptor cell types. Microvillous cells appeared largely unaffected by CuSO 4 treatment. Cilia in ciliated receptor neurones, however, appeared damaged one day post-treatment and were virtually absent after three days but had begun to recover after 14 days. Together, these results support the hypothesis that microvillous receptor neurones detect amino acids whereas ciliated receptor neurones were not functional and are responsible for detection of social stimuli (bile acids and pheromones). Furthermore, differences in sensitivity to copper may be due to different transduction pathways in the different cell types.
BMC Genomics, 2008
Background The goldfish (Carassius auratus) uses steroids and prostaglandins as pheromone cues at... more Background The goldfish (Carassius auratus) uses steroids and prostaglandins as pheromone cues at different stages of the reproductive cycle to facilitate spawning synchronization. Steroid progestin pheromone binding has been detected in goldfish olfactory membranes but the receptors responsible for this specific binding remain unknown. In order to shed some light on the olfactory epithelium transcriptome and search for possible receptor candidates a large set of EST from this tissue were analysed and compared to and combined with a similar zebrafish (Danio rerio) resource. Results We generated 4,797 high quality sequences from a normalized cDNA library of the goldfish olfactory epithelium, which were clustered in 3,879 unique sequences, grouped in 668 contigs and 3,211 singletons. BLASTX searches produced 3,243 significant (E-value < e-10) hits and Gene Ontology (GO) analysis annotated a further 1,223 of these genes (37.7%). Comparative analysis with zebrafish olfactory epitheli...
Cell and Tissue Biology, 2019
The pharmacokinetics of two substances containing recombinant insulin-like growth factor 1 (IGF1)... more The pharmacokinetics of two substances containing recombinant insulin-like growth factor 1 (IGF1) as an active substance was considered. The first substance (IGF 1) contains the actual recombinant IGF1, and the second (IGF 2) contains IGF1, which is translated from plasmid DNA encoding the IGF-1 gene for this protein. It was established that, with intramuscular administration of IGF 1 , the delay time for its entry into the bloodstream is 1.5-2 h, while that with IGF 2 is 24-25 h. This indicates the presence of various mechanisms of accumulation of these substances in the systemic circulation. The maximum concentration of IGF 1 in the blood was determined 5 h after administration and that of IGF 2 was determined 125 h after administration. The maximum concentrations of these substances are comparable to each other. The concentration of IGF 1 in the blood decreases to its initial value 12 h after its administration, and the concentration of IGF 2 does so after 216 h. The clearance parameters (Cl) and elimination constants (Kel) of the considered substances also had significant differences, which confirms the presence of fast and slow dynamics of the decrease of their maximum concentrations after intramuscular injection. The different dynamics of the accumulation of substances in the blood and their elimination from the bloodstream after administration, as well as the different values of the area parameters under the pharmacokinetic curve (AUC t , AUC ∞) demonstrate that IGF 2 has been in the systemic circulation for a longer time than IGF 1. This is essential for the appearance and severity of pharmacodynamic effects.
Journal of Applied Ichthyology, 2006
Reproductive toxicology (Elmsford, N.Y.), Dec 23, 2017
Earlier we showed that asymmetric methylation of sister chromatids (AMSC) was a specific characte... more Earlier we showed that asymmetric methylation of sister chromatids (AMSC) was a specific characteristic of differentiation potency, and supposed that AMSC could be a useful marker of environmental impact connected with differentiation and/or dedifferentiation. Here we investigated the level of AMSC in chromosomes and the nuclei methylation in mouse preimplantation and postimplantation embryos, in comparison with the undifferentiated cells of mouse embryonal carcinoma cell line F9, and human differentiated HEK293 cells upon BPA influence. We found that exposure of mouse preimplantation embryos to BPA caused a significant decrease in the level of AMSC in chromosomes and the nuclei methylation. The BPA exposure of potentially differentiating F9 cells had no any influence on DNA methylation in nuclei but significantly decreased the number of AMSC. The level of DNA methylation and AMSC in HEK293 cells were not also changed. These data indicate that BPA exerts significant influence on dif...
Medical academic journal, 2018
MemoriAl plate unveiled in memory of staff of the Institute of Experimental Medicine who were vic... more MemoriAl plate unveiled in memory of staff of the Institute of Experimental Medicine who were victims of political repression
Medical academic journal, 2018
This study aimed to develop a method for producing human recombinant insulin-like growth factor (... more This study aimed to develop a method for producing human recombinant insulin-like growth factor (IGF-1) based on a prokaryotic expression system and to characterize the highly purified protein. To achieve the study’s goal, the following methods were conducted: we performed automated chemical synthesis of DNA, constructed the expression plasmid, obtained Escherichia coli cell-producers of human recombinant IGF-1, cultivated the obtained producer cells with the induction of recombinant protein synthesis by isopropyl-β-D-1-thiogalactopyranoside and lactose, and purified human recombinant IGF-1 with affinity and cation exchange chromatography. The recombinant protein IGF-1 forms inclusion bodies during synthesis in Escherichia coli BL21 cells that contain plasmid pET28-IGF-1. Purified recombinant protein was obtained with a purity of 98% using affinity and cation exchange chromatography methods. The protein yield was 6 mg of human recombinant IGF-1 from 1 g of raw biomass. The resulting...
BioMetals, 2018
Among the properties of lactoferrin (LF) are bactericidal, antianemic, immunomodulatory, antitumo... more Among the properties of lactoferrin (LF) are bactericidal, antianemic, immunomodulatory, antitumour, antiphlogistic effects. Previously we demonstrated its capacity to stabilize in vivo HIF-1alpha and HIF-2-alpha, which are redox-sensitive multiaimed transcription factors. Various tissues of animals receiving recombinant human LF (rhLF) responded by expressing the HIF-1-alpha target genes, hence such proteins as erythropoietin (EPO), ceruloplasmin, etc. were synthesized in noticeable amounts. Among organs in which EPO synthesis occurred were brain, heart, spleen, liver, kidneys and lungs. Other researchers showed that EPO can act as a protectant against severe brain injury and status epilepticus in rats. Therefore, we tried rhLF as a protector against the severe neurologic disorders developed in rats, such as the rotenone-induced model of Parkinson's disease and experimental autoimmune encephalomyelitis as a model of multiple sclerosis, and observed its capacity to mitigate the grave symptoms. Moreover, an
Metallomics, 2017
Ceruloplasmin (Cp) is a copper-containing multifunctional oxidase of plasma. New rat Cp features ... more Ceruloplasmin (Cp) is a copper-containing multifunctional oxidase of plasma. New rat Cp features were discovered by X-ray analysis.
BioMetals, 2016
Strongly pronounced argyrosis caused by adding AgCl to the feed of laboratory rats efficiently mi... more Strongly pronounced argyrosis caused by adding AgCl to the feed of laboratory rats efficiently mimics the deficiency of ceruloplasmin (CP) ferroxidase activity. Bringing the concentration of AgCl in the feedstuff of lactating rats to 250 mg % and keeping their progeny (Ag-rats) for 3 months on the same silvercontaining feed provided the serum iron content 1.4 times lower than that in the control group. Besides, the ferroxidase activity of CP dropped to zero. In CP purified from sera of Ag-rats two copper ions were substituted with two silver ions. Using rat models of both posthemorrhagic and hemolytic anemia we showed that the deficiency of CP ferroxidase activity in Ag-rats affects the iron content in serum, though does not prevent the recovery of hemoglobin level accompanied by exhaustion of iron caches in liver and spleen. When apolactoferrin (apo-LF) was administered to Ag-rats suffering from either post-hemorrhagic or hemolytic anemia, both hemoglobin and serum iron were restored more rapidly than in the control animals. In independent experiments Ag-rats were compared with those fed on regular diet and the former displayed a prolonged 3-day stabilization of hypoxia-inducible factors 1 and 2 alpha (HIF-1a and HIF-2a) along with an increased serum concentration of erythropoietin. Introduction to Ag-rats of active CP separately or together with apo-LF reduced that effect to 1 day only. It is concluded that saturation of apo-LF with iron, provided by active CP, can strongly affect its protective capacity. Keywords Iron metabolism Á Ferroxidase Á Ceruloplasmin Á Lactoferrin Á Transferrin Á Erythropoietin Á Post-hemorrhagic anemia Á Hemolytic anemia Á Hypoxia-inducible factor Abbreviations Ag-rats Rats with AgCl-diet Apo-LF Apo-form LF CP Ceruloplasmin EPO Erythropoietin HIF-1a Hypoxia-inducible factor-1 alpha HIF-2a Hypoxia-inducible factor-2 alpha LF Lactoferrin TF Transferrin
Journal of Applied Ichthyology, 2006
Summary Sturgeon oocytes, like those of other vertebrates, are arrested in early meiosis and requ... more Summary Sturgeon oocytes, like those of other vertebrates, are arrested in early meiosis and require a steroid or steroids to trigger meiosis resumption or maturation. The identity of the maturation-inducing steroid in sturgeons has present study was designed to investigate in sterlet the follicular secretion and relative blood changes of free and conjugated 17, 20βP, 20βS and of the androgens T, 11KT, in response to hormonal treatment and to determine the relative potencies of the progestagens, 17, 20β-P, 20βS, P4 and S, in inducing oocyte final ...
Environmental Science & Technology, 2009
Exposure of olfactory epithelium to environmentally relevant concentrations of copper disrupts ol... more Exposure of olfactory epithelium to environmentally relevant concentrations of copper disrupts olfaction in fish. To examine the dynamics of recovery at both functional and morphological levels after acute copper exposure, unilateral exposure of goldfish olfactory epithelia to 100 µM CuSO 4 (10 min) was followed by electro-olfactogram (EOG) recording and scanning electron microscopy. Sensitivity to amino acids (L-arginine and L-serine), generally considered food-related odorants, recovered most rapidly (three days), followed by that to catecholamines (3-O-methoxytyramine), bile acids (taurolithocholic acid) and the steroid pheromone, 17,20-dihydroxy-4-pregnen-3-one 20-sulfate, which took 28 days to reach full recovery. Sensitivity to the postovulatory pheromone prostaglandin F 2R had not fully recovered even at 28 days. These changes in sensitivity were correlated with changes in the recovery of ciliated and microvillous receptor cell types. Microvillous cells appeared largely unaffected by CuSO 4 treatment. Cilia in ciliated receptor neurones, however, appeared damaged one day post-treatment and were virtually absent after three days but had begun to recover after 14 days. Together, these results support the hypothesis that microvillous receptor neurones detect amino acids whereas ciliated receptor neurones were not functional and are responsible for detection of social stimuli (bile acids and pheromones). Furthermore, differences in sensitivity to copper may be due to different transduction pathways in the different cell types.
BMC Genomics, 2008
Background The goldfish (Carassius auratus) uses steroids and prostaglandins as pheromone cues at... more Background The goldfish (Carassius auratus) uses steroids and prostaglandins as pheromone cues at different stages of the reproductive cycle to facilitate spawning synchronization. Steroid progestin pheromone binding has been detected in goldfish olfactory membranes but the receptors responsible for this specific binding remain unknown. In order to shed some light on the olfactory epithelium transcriptome and search for possible receptor candidates a large set of EST from this tissue were analysed and compared to and combined with a similar zebrafish (Danio rerio) resource. Results We generated 4,797 high quality sequences from a normalized cDNA library of the goldfish olfactory epithelium, which were clustered in 3,879 unique sequences, grouped in 668 contigs and 3,211 singletons. BLASTX searches produced 3,243 significant (E-value < e-10) hits and Gene Ontology (GO) analysis annotated a further 1,223 of these genes (37.7%). Comparative analysis with zebrafish olfactory epitheli...