Pub Date : 2024-07-24DOI: 10.33647/2074-5982-20-2-110-122
N. S. Ogneva, M. S. Nesterov, D. Khvostov, N. V. Stankova, V. Karkischenko
In this work, we investigate the pharmacokinetics of a new anti-inflammatory hexapeptide registered under the name of Leutragin. The study was conducted on Svetlogorsk minipigs by intravenous and a single rectal administration of the drug in the form of a solution and suppositories at an equal dose of 10 mg. The shortest time to reach peak concentration was demonstrated with intravenous administration, with the Tmax being 30 min. The maximum concentration (Cmax) when administering Leutragin in a suppository form was 141.37 ng/g. This concentration was achieved at the Tmax of 90 min, following which Leutragin remained in the bloodstream for 2.5 h. The absolute bioavailability of Leutragin in the suppository and solution form was 59.6% and 70.03%, respectively. The peak concentration of Leutragin under its rectal administration occurred at 150 min, following with the drug remained in the bloodstream for 4 h.
{"title":"Comparative Pharmacokinetics Study of the Leutragin Peptide Drug in Svetlogorsk Minipig Blood Serum after Single Administration","authors":"N. S. Ogneva, M. S. Nesterov, D. Khvostov, N. V. Stankova, V. Karkischenko","doi":"10.33647/2074-5982-20-2-110-122","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-110-122","url":null,"abstract":"In this work, we investigate the pharmacokinetics of a new anti-inflammatory hexapeptide registered under the name of Leutragin. The study was conducted on Svetlogorsk minipigs by intravenous and a single rectal administration of the drug in the form of a solution and suppositories at an equal dose of 10 mg. The shortest time to reach peak concentration was demonstrated with intravenous administration, with the Tmax being 30 min. The maximum concentration (Cmax) when administering Leutragin in a suppository form was 141.37 ng/g. This concentration was achieved at the Tmax of 90 min, following which Leutragin remained in the bloodstream for 2.5 h. The absolute bioavailability of Leutragin in the suppository and solution form was 59.6% and 70.03%, respectively. The peak concentration of Leutragin under its rectal administration occurred at 150 min, following with the drug remained in the bloodstream for 4 h.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"43 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141807755","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}
Pub Date : 2024-07-24DOI: 10.33647/2074-5982-20-2-95-109
O. V. Alimkina, N. V. Petrova, N. V. Stankova, Y. Fokin, E. S. Glotova, N. А. Laryushina, I. A. Vasil’eva
This article presents the results of 10-year research studies conducted using minipigs at the Scientific Center of Biomedical Technologies. Comparisons with the most significant laboratory animals are presented. Prospects for involving minipigs in various biomedical manipulations as an alternative to monkeys, whose use is restricted, are shown.
{"title":"Minipigs as Preferred Laboratory Animals for Extrapolation of Biomedical Research Data to Humans","authors":"O. V. Alimkina, N. V. Petrova, N. V. Stankova, Y. Fokin, E. S. Glotova, N. А. Laryushina, I. A. Vasil’eva","doi":"10.33647/2074-5982-20-2-95-109","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-95-109","url":null,"abstract":"This article presents the results of 10-year research studies conducted using minipigs at the Scientific Center of Biomedical Technologies. Comparisons with the most significant laboratory animals are presented. Prospects for involving minipigs in various biomedical manipulations as an alternative to monkeys, whose use is restricted, are shown.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"93 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141808133","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}
Pub Date : 2024-07-23DOI: 10.33647/2074-5982-20-2-21-31
V. Karkischenko, V. Ezerskiy, E. Koloskova, M. S. Nesterov
Transgenic humanized animals are increasingly in demand for biomedical research and pharmacological testing. More and more lines of transgenic animals are being created, including those with knockout of their own genes. There is an urgent need for an evidence base for the integration of a transgene, its expression, determination of the knockout state of its own gene at the molecular genetic level, detection of translation of the target protein in different organs and tissues, proof for the absence of protein synthesis (or its non-functionality), the gene of which has been modified. This requires highly specific reagents, proteins and antibodies to them in particular, the vast majority of which are presented by foreign manufacturers. The task was set to identify mouse and human β2-microglobulin in protein fractions of organs and tissues of transgenic and knockout mice of several HLA lines created in recent years at the Scientific Center of Biomedical Technologies, Russia. At the first stage of our research, recombinant E. coli producing strains were obtained.
生物医学研究和药理测试对转基因人源化动物的需求越来越大。越来越多的转基因动物品系正在诞生，包括那些自身基因被敲除的转基因动物。转基因的整合、表达、在分子基因水平上确定自身基因的敲除状态、检测目标蛋白在不同器官和组织中的翻译、证明被修饰基因不合成蛋白质（或无功能），这些都急需一个证据基础。这需要高度特异性的试剂、蛋白质和抗体，尤其是这些试剂、蛋白质和抗体，而绝大多数试剂、蛋白质和抗体都是由国外制造商提供的。俄罗斯生物医学技术科学中心（Scientific Center of Biomedical Technologies）的任务是在近几年培育出的几种 HLA 系转基因小鼠和基因敲除小鼠的器官和组织的蛋白质部分中鉴定小鼠和人类的 β2-微球蛋白。在研究的第一阶段，我们获得了重组大肠杆菌生产菌株。
{"title":"Preparation of Differentiated Recombinant Human β2-Microglobulin and Mouse β2-Microglobulin Proteins for its Detection in Class I HLA Chimeric Molecules","authors":"V. Karkischenko, V. Ezerskiy, E. Koloskova, M. S. Nesterov","doi":"10.33647/2074-5982-20-2-21-31","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-21-31","url":null,"abstract":"Transgenic humanized animals are increasingly in demand for biomedical research and pharmacological testing. More and more lines of transgenic animals are being created, including those with knockout of their own genes. There is an urgent need for an evidence base for the integration of a transgene, its expression, determination of the knockout state of its own gene at the molecular genetic level, detection of translation of the target protein in different organs and tissues, proof for the absence of protein synthesis (or its non-functionality), the gene of which has been modified. This requires highly specific reagents, proteins and antibodies to them in particular, the vast majority of which are presented by foreign manufacturers. The task was set to identify mouse and human β2-microglobulin in protein fractions of organs and tissues of transgenic and knockout mice of several HLA lines created in recent years at the Scientific Center of Biomedical Technologies, Russia. At the first stage of our research, recombinant E. coli producing strains were obtained.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"76 24","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141812580","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}
Pub Date : 2024-07-23DOI: 10.33647/2074-5982-20-2-66-94
N. Karkischenko, Y. Fokin, S. Kharitonov
A systematic study of γ-oscillations was carried out using rats with chronically implanted electrodes in the proreal gyrus, somatosensory cortex, dorsal hippocampus, and hypothalamus. Brain electrograms (BE) were recorded and investigated using an original software and hardware module. Linear diagrams were constructed using a QMS17 device in a frequency range of 60–250 Hz or greater. A mathematical analysis, normalization, and rationing of the series of γ-rhythms under the action of gamma-aminobutyric acid (GABA), acetylcholine (ACC), and insulin relative to similar background series were performed by double discrete-time Fourier transform and double angle arctangent function, which allowed us to extract relevant information from extremely small (1–2 μV) values of γ-oscillations. The accumulation of the substances under study was achieved by introducing the Aminalon (GABA), Galantamine (ACC), and liposomal Insulin pharmaceuticals. The plasma concentrations of the studied drugs were verified by HPLC and mathematical modeling. The normalized BE (NBE) reflected the intracentral mechanisms of action of the tested drugs, which were characterized by a stable picture in the resting state of the animals and under the action of Aminalon, Galantamine, and Insulin at the peak of their plasma concentrations (according to pharmacokinetic parameters). The γ-activity of the brain is maintained at the systemic level. Blockade of γ-oscillations in the frontal pole leads to their activation in the associated brain structures: the hypo-thalamus, reticular formation, caudate nucleus, etc. Under the influence of Aminalon, the total depressive effects were observed over the entire analyzed range in the posterior nucleus of the hypothalamus and proreal gyrus, as well as activating effects in the frequency range 60–75 Hz in the anterior suprasylvian gyrus. Under the action of Galantamine, partial depressive effects in the hippocampus and hypothalamus were observed at frequencies of about 60–65, 95–105, and 150 Hz. Under the action of liposomal Insulin, partial activating effects were noted in the anterior suprasylvian gyrus and in the dorsal hippocampus in the frequency range of 60–85 Hz.
{"title":"System Normalized Gamma Oscillations of Brain Structures: Pharmacological Analysis of Neurochemical and Metabolic Processes","authors":"N. Karkischenko, Y. Fokin, S. Kharitonov","doi":"10.33647/2074-5982-20-2-66-94","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-66-94","url":null,"abstract":"A systematic study of γ-oscillations was carried out using rats with chronically implanted electrodes in the proreal gyrus, somatosensory cortex, dorsal hippocampus, and hypothalamus. Brain electrograms (BE) were recorded and investigated using an original software and hardware module. Linear diagrams were constructed using a QMS17 device in a frequency range of 60–250 Hz or greater. A mathematical analysis, normalization, and rationing of the series of γ-rhythms under the action of gamma-aminobutyric acid (GABA), acetylcholine (ACC), and insulin relative to similar background series were performed by double discrete-time Fourier transform and double angle arctangent function, which allowed us to extract relevant information from extremely small (1–2 μV) values of γ-oscillations. The accumulation of the substances under study was achieved by introducing the Aminalon (GABA), Galantamine (ACC), and liposomal Insulin pharmaceuticals. The plasma concentrations of the studied drugs were verified by HPLC and mathematical modeling. The normalized BE (NBE) reflected the intracentral mechanisms of action of the tested drugs, which were characterized by a stable picture in the resting state of the animals and under the action of Aminalon, Galantamine, and Insulin at the peak of their plasma concentrations (according to pharmacokinetic parameters). The γ-activity of the brain is maintained at the systemic level. Blockade of γ-oscillations in the frontal pole leads to their activation in the associated brain structures: the hypo-thalamus, reticular formation, caudate nucleus, etc. Under the influence of Aminalon, the total depressive effects were observed over the entire analyzed range in the posterior nucleus of the hypothalamus and proreal gyrus, as well as activating effects in the frequency range 60–75 Hz in the anterior suprasylvian gyrus. Under the action of Galantamine, partial depressive effects in the hippocampus and hypothalamus were observed at frequencies of about 60–65, 95–105, and 150 Hz. Under the action of liposomal Insulin, partial activating effects were noted in the anterior suprasylvian gyrus and in the dorsal hippocampus in the frequency range of 60–85 Hz.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"120 35","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141811735","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}
Pub Date : 2024-07-23DOI: 10.33647/2074-5982-20-2-53-65
N. Karkischenko, V. Ezerskiy, O. B. Zhukova, E. Koloskova, N. V. Petrova
Highly specific reagents, i.e., proteins and antibodies to them, are the necessary components of systems for verifying the effectiveness of transgenic/knockout animal biomodels. In particular, the identification of mouse and human β2-microglobulin in the protein fractions of organs and tissues of transgenic and β2m knockout mice of several HLA lines, which have been created in recent years at the Scientific Center of Biomedical Technologies of the FMBA of Russia, is the most important stage of their certification. At the first stage of our research, E. coli producing strains of recombinant mouse and human β2-microglobulin (mβ2mg and hβ2mg) were obtained, the proteins were isolated and purified. At the next stage of the work, affine sorbents with immobilized mβ2mg and hβ2mg were obtained. To increase the species specificity of the serum, “rabbit-anti-hβ2mg” were depleted against the recombinant protein mβ2mg, and, conversely, “rabbit-anti-mβ2mg” were depleted against the recombinant protein hβ2mg. Highly specific antibodies were purified from depleted sera using affinity sorbents. Using dot- and western-blotting methods оn the example of depleted and affinity-purified rabbit-anti-hβ2mg antibodies, a significant increase in their specificity relative to hβ2mg was shown.
{"title":"Increasing, the Specificity of Polyclonal Antibodies to Human and Mouse β2-Microglobulin as an Alternative to the Use of Monoclonal Antibodies in Immunological Analysis","authors":"N. Karkischenko, V. Ezerskiy, O. B. Zhukova, E. Koloskova, N. V. Petrova","doi":"10.33647/2074-5982-20-2-53-65","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-53-65","url":null,"abstract":"Highly specific reagents, i.e., proteins and antibodies to them, are the necessary components of systems for verifying the effectiveness of transgenic/knockout animal biomodels. In particular, the identification of mouse and human β2-microglobulin in the protein fractions of organs and tissues of transgenic and β2m knockout mice of several HLA lines, which have been created in recent years at the Scientific Center of Biomedical Technologies of the FMBA of Russia, is the most important stage of their certification. At the first stage of our research, E. coli producing strains of recombinant mouse and human β2-microglobulin (mβ2mg and hβ2mg) were obtained, the proteins were isolated and purified. At the next stage of the work, affine sorbents with immobilized mβ2mg and hβ2mg were obtained. To increase the species specificity of the serum, “rabbit-anti-hβ2mg” were depleted against the recombinant protein mβ2mg, and, conversely, “rabbit-anti-mβ2mg” were depleted against the recombinant protein hβ2mg. Highly specific antibodies were purified from depleted sera using affinity sorbents. Using dot- and western-blotting methods оn the example of depleted and affinity-purified rabbit-anti-hβ2mg antibodies, a significant increase in their specificity relative to hβ2mg was shown.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"120 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141811869","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}
Pub Date : 2024-07-23DOI: 10.33647/2074-5982-20-2-45-52
V. Karkischenko, A. G. Berzina, I. A. Pomytkin, E. S. Glotova, M. A. Savina, D. V. Petrov, L. A. Taboyakova, L. A. Bolotskih, I. A. Vasil’eva
The introduction of a transgene can impact negatively the functioning of vital systems in biomodels. We carried out a comparative analysis of the immune response of mice of the HLA-A*02:01 humanized transgenic line, mice with mouse β2-microglobulin gene knockout, and wild-type mice to the introduction of horse immunoglobulin as an antigen. The biomodel lines were created at the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia. The maximum immune response was achieved on the 30th day from the onset of immunization in animals of the HLA-A*02:01 line and wild-type mice. Antibody titers in these groups increased sharply and approached 1:8,000,000 and 1:4,000,000, respectively. This indicates that genome modification in HLA-A*02:01 transgenic humanized mice did not affect functioning of the immune system. No similar dynamics of the increase in antibody titers was observed in the mice line with mouse β2-microglobulin gene knockout. On the 7th and 30th day, the antibody titer in this group increased to a value of 1:400 and 1:6,400, respectively. The weak immune response in mice with mouse β2-microglobulin gene knockout confirms the undeniably important role of this protein in immune response formation.
{"title":"Immune Response in HLA-A*02:01 Transgenic Humanized Mice to the Introduction of Horse IgG Antigen","authors":"V. Karkischenko, A. G. Berzina, I. A. Pomytkin, E. S. Glotova, M. A. Savina, D. V. Petrov, L. A. Taboyakova, L. A. Bolotskih, I. A. Vasil’eva","doi":"10.33647/2074-5982-20-2-45-52","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-45-52","url":null,"abstract":"The introduction of a transgene can impact negatively the functioning of vital systems in biomodels. We carried out a comparative analysis of the immune response of mice of the HLA-A*02:01 humanized transgenic line, mice with mouse β2-microglobulin gene knockout, and wild-type mice to the introduction of horse immunoglobulin as an antigen. The biomodel lines were created at the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia. The maximum immune response was achieved on the 30th day from the onset of immunization in animals of the HLA-A*02:01 line and wild-type mice. Antibody titers in these groups increased sharply and approached 1:8,000,000 and 1:4,000,000, respectively. This indicates that genome modification in HLA-A*02:01 transgenic humanized mice did not affect functioning of the immune system. No similar dynamics of the increase in antibody titers was observed in the mice line with mouse β2-microglobulin gene knockout. On the 7th and 30th day, the antibody titer in this group increased to a value of 1:400 and 1:6,400, respectively. The weak immune response in mice with mouse β2-microglobulin gene knockout confirms the undeniably important role of this protein in immune response formation.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"18 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141813787","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}
Pub Date : 2024-07-23DOI: 10.33647/2074-5982-20-2-32-44
V. Karkischenko, A. G. Berzina, N. V. Petrova, I. A. Pomytkin, E. S. Glotova, D. V. Petrov, L. A. Taboyakova, L. A. Bolotskih, N. А. Laryushina
Human leukocyte antigen plays a primary role in the formation of immune response and pathogenesis of diseases of various etiologies, including the development of negative side effects induced by pharmacological agents. Modern pharmacosafety standards require improvement of existing test systems to conduct high-quality preclinical studies. A number of humanized transgenic mouse lines with hybrid HLA I class molecules on the cell surface, which correspond to the human allelic variants HLA-A*02:01, HLA-B*07:02, and HLA-C*07:02, were developed at the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia. In this article, we present experimental data on quantitative determination of β2-microglobulin protein and HLA by the “sandwich” ELISA method in mice with different alleles of HLA I class genes. The results obtained confirm the presence of target functional proteins (transgenicity) in humanized transgenic mice, which is consistent with our previous data obtained when determining the primary sequence of the transgene using Sanger sequencing. We also discuss the scientific and practical significance of such biomodels, as well as the scope of their application.
人类白细胞抗原在免疫反应的形成和各种病因疾病的发病机制中发挥着主要作用，包括药理制剂诱发的负面副作用。现代药物安全标准要求改进现有的试验系统，以开展高质量的临床前研究。俄罗斯联邦医学和生物局生物医学技术科学中心开发了一些细胞表面具有混合 HLA I 类分子的人源化转基因小鼠品系，这些品系对应于人类等位基因变体 HLA-A*02:01、HLA-B*07:02 和 HLA-C*07:02。本文介绍了采用 "夹心 "ELISA 法定量检测 HLA I 类基因不同等位基因小鼠体内 β2-微球蛋白和 HLA 的实验数据。结果证实人源化转基因小鼠体内存在目标功能蛋白（转基因性），这与我们之前利用桑格测序法确定转基因主序列时获得的数据一致。我们还讨论了此类生物模型的科学和实用意义及其应用范围。
{"title":"Evidence for the Presence of β2m hom Target Proteins and HLA in Humanized Transgenic Mice of HLA-A*02:01, HLA-B*07:02, and HLA-C*07:02 Lines","authors":"V. Karkischenko, A. G. Berzina, N. V. Petrova, I. A. Pomytkin, E. S. Glotova, D. V. Petrov, L. A. Taboyakova, L. A. Bolotskih, N. А. Laryushina","doi":"10.33647/2074-5982-20-2-32-44","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-32-44","url":null,"abstract":"Human leukocyte antigen plays a primary role in the formation of immune response and pathogenesis of diseases of various etiologies, including the development of negative side effects induced by pharmacological agents. Modern pharmacosafety standards require improvement of existing test systems to conduct high-quality preclinical studies. A number of humanized transgenic mouse lines with hybrid HLA I class molecules on the cell surface, which correspond to the human allelic variants HLA-A*02:01, HLA-B*07:02, and HLA-C*07:02, were developed at the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia. In this article, we present experimental data on quantitative determination of β2-microglobulin protein and HLA by the “sandwich” ELISA method in mice with different alleles of HLA I class genes. The results obtained confirm the presence of target functional proteins (transgenicity) in humanized transgenic mice, which is consistent with our previous data obtained when determining the primary sequence of the transgene using Sanger sequencing. We also discuss the scientific and practical significance of such biomodels, as well as the scope of their application.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"16 12","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141813486","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}
Pub Date : 2024-07-22DOI: 10.33647/2074-5982-20-2-8-20
N. Karkischenko, N. V. Petrova, V. Slobodenyuk, E. Koloskova, N. А. Laryushina, I. A. Vasil’eva, D. V. Petrov, L. A. Bolotskih, M. A. Savina
Several humanized transgenic lines of biomodel mice containing an integrated variable human gene of the main histocompatibility complex (MHC) have been created at the Federal State Budgetary Scientific and Scientific Research Institute of the FMBA of Russia. These include HLA-A*02:01, HLA-B*07:02 and HLA-C*07:02. The lines were created by microinjection of a linear fragment of a genetically engineered structure (GES) into the male pronucleus of zygotes, followed by the transfer of potentially modified embryos into the reproductive tract to pseudo-pregnant female recipients. The created GES encodes a chimeric molecule of the MHC of class I on the cell surface, consisting of the α1-, α2-domains of human HLA, and the α3-domain of the mouse H-2K complex, stabilized by human β2-microglobulin connected by a glycine serine linker with the α1-domain of HLA [1–5]. The created biomodels can be successfully used to solve a wide range of research tasks, including studies of immune reactions, infectious, autoimmune and oncological diseases, as well as the development and testing of vaccines in the field of pharmacosafety and immunogenicity. This article presents theoretical information on the genetic polymorphism of the studied gene in the human genome, as well as experimental data on the transgenic lines of biomodels created by the authors and the results of comparing the allele-specific site in the obtained animal lines. The analysis was performed by Sanger sequencing on a cDNA matrix.
{"title":"Evidence of Trangency and Humanization in Mice Obtained at the SCBMT of FMBA of Russia by Sanger Sequencing Method","authors":"N. Karkischenko, N. V. Petrova, V. Slobodenyuk, E. Koloskova, N. А. Laryushina, I. A. Vasil’eva, D. V. Petrov, L. A. Bolotskih, M. A. Savina","doi":"10.33647/2074-5982-20-2-8-20","DOIUrl":"https://doi.org/10.33647/2074-5982-20-2-8-20","url":null,"abstract":"Several humanized transgenic lines of biomodel mice containing an integrated variable human gene of the main histocompatibility complex (MHC) have been created at the Federal State Budgetary Scientific and Scientific Research Institute of the FMBA of Russia. These include HLA-A*02:01, HLA-B*07:02 and HLA-C*07:02. The lines were created by microinjection of a linear fragment of a genetically engineered structure (GES) into the male pronucleus of zygotes, followed by the transfer of potentially modified embryos into the reproductive tract to pseudo-pregnant female recipients. The created GES encodes a chimeric molecule of the MHC of class I on the cell surface, consisting of the α1-, α2-domains of human HLA, and the α3-domain of the mouse H-2K complex, stabilized by human β2-microglobulin connected by a glycine serine linker with the α1-domain of HLA [1–5]. The created biomodels can be successfully used to solve a wide range of research tasks, including studies of immune reactions, infectious, autoimmune and oncological diseases, as well as the development and testing of vaccines in the field of pharmacosafety and immunogenicity. This article presents theoretical information on the genetic polymorphism of the studied gene in the human genome, as well as experimental data on the transgenic lines of biomodels created by the authors and the results of comparing the allele-specific site in the obtained animal lines. The analysis was performed by Sanger sequencing on a cDNA matrix.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"17 23","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141816309","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}
Pub Date : 2024-04-08DOI: 10.33647/2074-5982-20-1-52-61
A. Zharikov, S. O. Filinova, O. Mazko, I. P. Bobrov, O. Makarova, A. Kalnitsky
The article presents the results of a study into the effect of carnosine on oxidative damage to the kidneys in experimental diabetes mellitus. The experiment was carried out using two groups of Wistar rats: control (n=8) and experimental (n=11). In both groups, streptozotocin-induced diabetes mellitus was simulated for eight weeks. Experimental animals were intragastrically injected with carnosine (15 mg/kg) from weeks 4 to 8. The concentration of glucose, protein and creatinine excretion in urine were determined. At the end of eight weeks, the kidneys were removed from the rats to determine the indicators of oxidative stress severity (concentration of thiobarbiturate-reactive products, total antioxidant activity, activity of catalase, superoxide dismutase, glutathione peroxidase) and to conduct morphometry of the size of the renal glomeruli, the area of the vascular bed, capillaries and mesangium in the glomeruli, the number of podocytes. A comparison with the control showed the use of carnosine led to a 1.5-fold decrease in the concentration of thiobarbiturate-reactive products (p<0.001), a 2.2-fold increase in the total antioxidant activity (p<0.001), and a 1.2-fold increase in catalase activity (p=0.039). The area of the renal glomeruli and the mesangium in this group decreased by 1.6 times (p<0.001 and p=0.04, respectively). The total area of blood flow increased by 2.4 times (p<0.001), the area of one capillary, and the number of podocytes in the glomerulus increased by 1.9 times (p<0.001 and p=0.001). A 3.5-fold decrease in protein concentration in urine was also noted (p=0.007). Therefore, inhibition of the formation of advanced glycation end products by carnosine in experimental diabetes mellitus attenuates oxidative damage to the kidneys. This is evidenced by a decrease in proteinuria, an increase in the number of podocytes, a decrease in the area of the renal glomeruli, and an improvement in the condition of the glomerular vascular system.
{"title":"Effect of Carnosine on Oxidative Damage to the Kidneys in Experinental Diabetes Mellitus","authors":"A. Zharikov, S. O. Filinova, O. Mazko, I. P. Bobrov, O. Makarova, A. Kalnitsky","doi":"10.33647/2074-5982-20-1-52-61","DOIUrl":"https://doi.org/10.33647/2074-5982-20-1-52-61","url":null,"abstract":"The article presents the results of a study into the effect of carnosine on oxidative damage to the kidneys in experimental diabetes mellitus. The experiment was carried out using two groups of Wistar rats: control (n=8) and experimental (n=11). In both groups, streptozotocin-induced diabetes mellitus was simulated for eight weeks. Experimental animals were intragastrically injected with carnosine (15 mg/kg) from weeks 4 to 8. The concentration of glucose, protein and creatinine excretion in urine were determined. At the end of eight weeks, the kidneys were removed from the rats to determine the indicators of oxidative stress severity (concentration of thiobarbiturate-reactive products, total antioxidant activity, activity of catalase, superoxide dismutase, glutathione peroxidase) and to conduct morphometry of the size of the renal glomeruli, the area of the vascular bed, capillaries and mesangium in the glomeruli, the number of podocytes. A comparison with the control showed the use of carnosine led to a 1.5-fold decrease in the concentration of thiobarbiturate-reactive products (p<0.001), a 2.2-fold increase in the total antioxidant activity (p<0.001), and a 1.2-fold increase in catalase activity (p=0.039). The area of the renal glomeruli and the mesangium in this group decreased by 1.6 times (p<0.001 and p=0.04, respectively). The total area of blood flow increased by 2.4 times (p<0.001), the area of one capillary, and the number of podocytes in the glomerulus increased by 1.9 times (p<0.001 and p=0.001). A 3.5-fold decrease in protein concentration in urine was also noted (p=0.007). Therefore, inhibition of the formation of advanced glycation end products by carnosine in experimental diabetes mellitus attenuates oxidative damage to the kidneys. This is evidenced by a decrease in proteinuria, an increase in the number of podocytes, a decrease in the area of the renal glomeruli, and an improvement in the condition of the glomerular vascular system.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"31 14","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140728079","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}
Pub Date : 2024-04-08DOI: 10.33647/2074-5982-20-1-62-72
M. Sergalieva, A. A. Tsibizova, M. Samotrueva
We investigate effects of glyproline-type neuropeptides on lipid and protein peroxidation in the hypothalamic brain region of rats under the conditions of experimental hyperthyroidism. The state of hyperthyroidism in animals was simulated by intragastric administration of sodium pentahydrate of L-thyroxine at a dose of 150 µg/kg for 21 days. The following experimental groups (n=10) were formed: 1) control group — intact animals (control); 2) animals treated with L-thyroxine sodium salt pentahydrate (hyperthyroidism); 3) animals treated with Thr-Lys-Pro-Arg-Pro-Gly-Pro (celank); 4) animals treated with Pro-GlyPro (doses of 87 and 33 µg/kg/day, respectively) intraperitoneally daily during 21 days starting one day after the last administration of sodium pentahydrate of L-thyroxine. The level of lipid peroxidation processes was assessed by the initial level of TBA-reactive products, spontaneous and ascorbate-dependent lipid peroxidation rates in hypothalamic tissue homogenate. Protein peroxidation products were determined by the reaction between oxidized amino acid residues of proteins and 2,4-dinitrophenylhydrazone. The enzymatic part of the antioxidant system of hypothalamic region was estimated by measuring the activity of superoxide dismutase and catalase. In the setting of Thr-Lys-Pro-Arg-Pro-Gly-Pro and Pro-Gly-Pro administration under experimental hyperthyroidism, the intensity of lipid and protein peroxidation processes was decreased, while the activity of antioxidant enzymes-superoxide dismutase and catalase was restored in the hypothalamic tissue. The experimental data obtained indicate that, under the conditions of experimental hyperthyroidism, Thr-Lys-Pro-Arg-Pro-Gly-Pro and Pro-Gly-Pro compounds exhibit an antioxidant and antiradical activity with respect to parameters of lipoperoxidation and oxidative modification of proteins, as well as with respect to enzymatic defense systems in the hypothalamic brain region of laboratory animals.
{"title":"Effects of Glyprolins on Lipid and Protein Peroxidation in the Hypothalamic Region in Experimental Hyperthyroidism","authors":"M. Sergalieva, A. A. Tsibizova, M. Samotrueva","doi":"10.33647/2074-5982-20-1-62-72","DOIUrl":"https://doi.org/10.33647/2074-5982-20-1-62-72","url":null,"abstract":"We investigate effects of glyproline-type neuropeptides on lipid and protein peroxidation in the hypothalamic brain region of rats under the conditions of experimental hyperthyroidism. The state of hyperthyroidism in animals was simulated by intragastric administration of sodium pentahydrate of L-thyroxine at a dose of 150 µg/kg for 21 days. The following experimental groups (n=10) were formed: 1) control group — intact animals (control); 2) animals treated with L-thyroxine sodium salt pentahydrate (hyperthyroidism); 3) animals treated with Thr-Lys-Pro-Arg-Pro-Gly-Pro (celank); 4) animals treated with Pro-GlyPro (doses of 87 and 33 µg/kg/day, respectively) intraperitoneally daily during 21 days starting one day after the last administration of sodium pentahydrate of L-thyroxine. The level of lipid peroxidation processes was assessed by the initial level of TBA-reactive products, spontaneous and ascorbate-dependent lipid peroxidation rates in hypothalamic tissue homogenate. Protein peroxidation products were determined by the reaction between oxidized amino acid residues of proteins and 2,4-dinitrophenylhydrazone. The enzymatic part of the antioxidant system of hypothalamic region was estimated by measuring the activity of superoxide dismutase and catalase. In the setting of Thr-Lys-Pro-Arg-Pro-Gly-Pro and Pro-Gly-Pro administration under experimental hyperthyroidism, the intensity of lipid and protein peroxidation processes was decreased, while the activity of antioxidant enzymes-superoxide dismutase and catalase was restored in the hypothalamic tissue. The experimental data obtained indicate that, under the conditions of experimental hyperthyroidism, Thr-Lys-Pro-Arg-Pro-Gly-Pro and Pro-Gly-Pro compounds exhibit an antioxidant and antiradical activity with respect to parameters of lipoperoxidation and oxidative modification of proteins, as well as with respect to enzymatic defense systems in the hypothalamic brain region of laboratory animals.","PeriodicalId":14837,"journal":{"name":"Journal Biomed","volume":"12 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140728605","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}
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