Pub Date : 2022-12-30DOI: 10.15407/biotech15.06.026
Y.B. Yanvarov
Surfactants are widely used in many areas of our life. However, synthetic surfactants have a serious negative impact on the environment. They do not decompose well and can accumulate in ecosystems. Microbial biosurfactants can be an alternative to synthetic surfactants. They are characterized by a diverse structure, stable at critical temperatures, pH and can be obtained from various renewable raw materials. Goal: analysis and generalization of the available information on the main characteristics and features of the synthesis of surface-active substances of microbial origin. Results. The article describes the structure of the most important groups of biosurfactants of microbial origin, such as rhamnolipids, trehalolipids, and sophorolipids. The main producers of biosurfactants, as well as the areas of their application were characterized. Information about the main ways of their biosynthesis is discussed. Special attention in the review is paid to factors that are essential for the cultivation of microorganisms - the main producers of biosurfactants.
{"title":"BIOSURFACTANTS: STRUCTURE, FUNCTIONS AND PRODUCTIONS","authors":"Y.B. Yanvarov","doi":"10.15407/biotech15.06.026","DOIUrl":"https://doi.org/10.15407/biotech15.06.026","url":null,"abstract":"Surfactants are widely used in many areas of our life. However, synthetic surfactants have a serious negative impact on the environment. They do not decompose well and can accumulate in ecosystems. Microbial biosurfactants can be an alternative to synthetic surfactants. They are characterized by a diverse structure, stable at critical temperatures, pH and can be obtained from various renewable raw materials. Goal: analysis and generalization of the available information on the main characteristics and features of the synthesis of surface-active substances of microbial origin. Results. The article describes the structure of the most important groups of biosurfactants of microbial origin, such as rhamnolipids, trehalolipids, and sophorolipids. The main producers of biosurfactants, as well as the areas of their application were characterized. Information about the main ways of their biosynthesis is discussed. Special attention in the review is paid to factors that are essential for the cultivation of microorganisms - the main producers of biosurfactants.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48220512","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 : 2022-12-30DOI: 10.15407/biotech15.06.061
Daria Korolova Korolova
The most accurate laboratory methods for thrombophilia diagnostics are based on the quantitative determination of the blood plasma specific markers that appear as a result of the coagulation cascade activation. Soluble fibrin and D-dimer belong to the main of the last ones. An alteration in the concentration of such markers can indicate thrombin concentration growth and the formation of soluble oligomeric fibrin. It should be pointed out that simultaneous detection of these markers can establish the correlation between the accumulation of soluble fibrin and fibrinolysis and nowadays is provided only by enzyme-linked immunoassay. Thus, the usage of immunodiagnostic test systems for the detection of thrombophilia markers is highly relevant today. The important components of immunodiagnostic test system are protein calibrators, the isolation standardization of which plays a key role for accurate construction of a calibration curve and obtaining objective results as a consequence. Aim. The objective of this study was to develop the soluble fibrin and D-dimer isolation methodology and its standardization for their further use as the protein calibrators for thrombophilia markers detecting immunodiagnostic test systems. Materials and Methods. Soluble fibrin and D-dimer were isolated from collected human blood by fibrinogen salting out with further fibrin polymerization with thrombin and hydrolysis with plasmin. Quality control of the obtained proteins was carried out using SDS-PAGE and turbidimetric measurements with further checking of the proteins as calibrators for the thrombophilia markers detecting immunoassay. Results. Obtained proteins meet the necessary specifications and can be used as calibrators for immunodiagnostic test systems. Soluble fibrin and D-dimer were checked by SDS-PAGE for the absence of impurities. Turbidimetric measurements showed the polymerization capability of the soluble fibrin and the inhibition of the polymerization by D-dimer. Conclusion. The standardized isolation methodology of soluble fibrin and D-dimer can be used to obtain protein calibrators for appropriate immunodiagnostic test systems.
{"title":"STANDARDIZATION OF THE PROTEIN CALIBRATORS ISOLATION METHODOLOGY FOR THROMBOPHILIA MARKERS DETECTING IMMUNODIAGNOSTIC TEST SYSTEMS","authors":"Daria Korolova Korolova","doi":"10.15407/biotech15.06.061","DOIUrl":"https://doi.org/10.15407/biotech15.06.061","url":null,"abstract":"The most accurate laboratory methods for thrombophilia diagnostics are based on the quantitative determination of the blood plasma specific markers that appear as a result of the coagulation cascade activation. Soluble fibrin and D-dimer belong to the main of the last ones. An alteration in the concentration of such markers can indicate thrombin concentration growth and the formation of soluble oligomeric fibrin. It should be pointed out that simultaneous detection of these markers can establish the correlation between the accumulation of soluble fibrin and fibrinolysis and nowadays is provided only by enzyme-linked immunoassay. Thus, the usage of immunodiagnostic test systems for the detection of thrombophilia markers is highly relevant today. The important components of immunodiagnostic test system are protein calibrators, the isolation standardization of which plays a key role for accurate construction of a calibration curve and obtaining objective results as a consequence. Aim. The objective of this study was to develop the soluble fibrin and D-dimer isolation methodology and its standardization for their further use as the protein calibrators for thrombophilia markers detecting immunodiagnostic test systems. Materials and Methods. Soluble fibrin and D-dimer were isolated from collected human blood by fibrinogen salting out with further fibrin polymerization with thrombin and hydrolysis with plasmin. Quality control of the obtained proteins was carried out using SDS-PAGE and turbidimetric measurements with further checking of the proteins as calibrators for the thrombophilia markers detecting immunoassay. Results. Obtained proteins meet the necessary specifications and can be used as calibrators for immunodiagnostic test systems. Soluble fibrin and D-dimer were checked by SDS-PAGE for the absence of impurities. Turbidimetric measurements showed the polymerization capability of the soluble fibrin and the inhibition of the polymerization by D-dimer. Conclusion. The standardized isolation methodology of soluble fibrin and D-dimer can be used to obtain protein calibrators for appropriate immunodiagnostic test systems.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42724693","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 : 2022-12-30DOI: 10.15407/biotech15.06.036
P. V. Beloshitsky, O. Klyuchko, Yu. M. Onopchuk
Investigations of the adaptation of living organisms/human body to various extreme factors are extremely important. Aim. To characterize and analyze the results of research of structural and functional interdependencies of organisms in extreme conditions. Methods. Comparative analysis of the registered biochemical, physiological characteristics of the body, mathematical modelling of underlying mechanisms on their basis, information and computer technologies. Results. Deviations of organisms’ functions during adaptation processes caused changes in some structures of organism. Significant role of quantitative and qualitative changes of the erythrocyte formation system in the reliability of organisms functioning in extreme conditions in highlands was confirmed. The changes in red and white blood cells reflected largely the relationships between the organisms’ reactivity and resistance. The dependences on degree of rarefaction of the air, mode of climbing, effects of athlete’s training, etc. were revealed. Adaptive hemolysis of erythrocytes, when the biologically active substances were released from blood cells and acted as messengers, were shown to be the triggers capable to change cell metabolism; they played significant roles in reliability of organisms functioning. The set of program models was developed. Results were applied successfully for training of athletes for high-altitude climbing. Conclusions. Results of the studies on the structural and functional interdependencies of organisms in extreme conditions were reviewed and analyzed. Results of mathematical modeling coincided with the results obtained in experiments and observations. In the process of adaptation to hypoxia human organism behaved likes an ultrastable system. Obtained results can be applied in practice.
{"title":"STRUCTURAL AND FUNCTIONAL INTERDEPENDENCES OF BIOLOGICAL ORGANISMS IN EXTREME CONDITIONS","authors":"P. V. Beloshitsky, O. Klyuchko, Yu. M. Onopchuk","doi":"10.15407/biotech15.06.036","DOIUrl":"https://doi.org/10.15407/biotech15.06.036","url":null,"abstract":"Investigations of the adaptation of living organisms/human body to various extreme factors are extremely important. Aim. To characterize and analyze the results of research of structural and functional interdependencies of organisms in extreme conditions. Methods. Comparative analysis of the registered biochemical, physiological characteristics of the body, mathematical modelling of underlying mechanisms on their basis, information and computer technologies. Results. Deviations of organisms’ functions during adaptation processes caused changes in some structures of organism. Significant role of quantitative and qualitative changes of the erythrocyte formation system in the reliability of organisms functioning in extreme conditions in highlands was confirmed. The changes in red and white blood cells reflected largely the relationships between the organisms’ reactivity and resistance. The dependences on degree of rarefaction of the air, mode of climbing, effects of athlete’s training, etc. were revealed. Adaptive hemolysis of erythrocytes, when the biologically active substances were released from blood cells and acted as messengers, were shown to be the triggers capable to change cell metabolism; they played significant roles in reliability of organisms functioning. The set of program models was developed. Results were applied successfully for training of athletes for high-altitude climbing. Conclusions. Results of the studies on the structural and functional interdependencies of organisms in extreme conditions were reviewed and analyzed. Results of mathematical modeling coincided with the results obtained in experiments and observations. In the process of adaptation to hypoxia human organism behaved likes an ultrastable system. Obtained results can be applied in practice.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44146174","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 : 2022-12-30DOI: 10.15407/biotech15.06.005
M. Kaniuk
The aim of the work was to review the literature data regarding the prospects for the creation and use of multifunctional fluorescent two-dye nanosystems, which enable investigating the distribution of fluorescent components with significant acceleration of the study and introduction of nanomedicines into practice. Special attention is paid to the use of two substances with hydrophobic and hydrophilic properties in one nanoparticle (NP), capable of penetrating a living cell. The method of fluorescence confocal microscopy enables observation of the nanoscale dynamics of distribution and stability of drugs over time. The concomitant use of doxorubicin (DOX) and curcumin (CUR) in single nanoparticle causes synergism in the action of medical drugs, and their own fluorescence makes it possible to use them as multifunctional fluorescent nanosystems. Results. Data from the literature indicate that the use of two or more fluorescent dyes provide an advantage over other, more expensive methods when studying the penetration and distribution of NPs in living samples. The use of nanocarriers is an effective way to significantly increase the bioavailability of those drugs, which are poorly soluble in water. A promising direction of nanomedicine is the creation of complex bio-compatible multifunctional nanomaterials based on several active drugs, with the simultaneous use of their enhancers and the strategy of active targeting. Such recent structures enable targeted and controlled penetration of medicinal compounds into the sites of localization of pathological processes, reducing the toxicity of drugs to normal cells. Conclusions. The use of the fluorescence microscopy method, as exemplified by the two dyes, DOX and CUR, enables to trace the stages of interaction of loaded DOX and CUR nanoparticles with cultured cells, and their release from NPs to determine their amount and localization in organelles cells.
{"title":"MULTIFUNCTIONAL NANOSYSTEMS BASED ON TWO FLUORESCENT DYES, DOXORUBICIN AND CURCUMIN","authors":"M. Kaniuk","doi":"10.15407/biotech15.06.005","DOIUrl":"https://doi.org/10.15407/biotech15.06.005","url":null,"abstract":"The aim of the work was to review the literature data regarding the prospects for the creation and use of multifunctional fluorescent two-dye nanosystems, which enable investigating the distribution of fluorescent components with significant acceleration of the study and introduction of nanomedicines into practice. Special attention is paid to the use of two substances with hydrophobic and hydrophilic properties in one nanoparticle (NP), capable of penetrating a living cell. The method of fluorescence confocal microscopy enables observation of the nanoscale dynamics of distribution and stability of drugs over time. The concomitant use of doxorubicin (DOX) and curcumin (CUR) in single nanoparticle causes synergism in the action of medical drugs, and their own fluorescence makes it possible to use them as multifunctional fluorescent nanosystems. Results. Data from the literature indicate that the use of two or more fluorescent dyes provide an advantage over other, more expensive methods when studying the penetration and distribution of NPs in living samples. The use of nanocarriers is an effective way to significantly increase the bioavailability of those drugs, which are poorly soluble in water. A promising direction of nanomedicine is the creation of complex bio-compatible multifunctional nanomaterials based on several active drugs, with the simultaneous use of their enhancers and the strategy of active targeting. Such recent structures enable targeted and controlled penetration of medicinal compounds into the sites of localization of pathological processes, reducing the toxicity of drugs to normal cells. Conclusions. The use of the fluorescence microscopy method, as exemplified by the two dyes, DOX and CUR, enables to trace the stages of interaction of loaded DOX and CUR nanoparticles with cultured cells, and their release from NPs to determine their amount and localization in organelles cells.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44424987","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 : 2022-10-01DOI: 10.15407/biotech15.05.064
Berfin Tugba Turak
Aim: Many genetic and environmental factors can be effective in the process of cancerization. Preventing the progression of leukemia may be possible by controlling the pathways involving mechanisms such as apoptosis and autophagy. When the literature is examined, there are studies showing the effects of various types of juniper on various cancer cell lines, including human chronic myeloid leukemia cells, but the signal pathways in which they act are not fully known. In this study, the anticancer effects of Juniperus oxycedrus extract on K-562 human chronic myeloid leukemia cells were investigated. Method: After the cells were treated with the Juniperus oxycedrus extract, cytotoxicity and gene expression analyzes were performed. Changes in the expression of Akt, the member of the PI3K/Akt/mTOR signaling pathway; caspase 3, which is one of the main effective genes in the pathways regulating apoptosis; and the apoptosis suppressor BCL-2 gene, which is an oncogene, were investigated. Results: According to the MTT test results, Juniperus oxycedrus extract showed over approximately 50% cell viability in K-562 cells at all doses. The most appropriate dose of Juniperus oxycedrus fruit extract in this research was determined as 50 µg/ mL considering cell viability. After the gene expression analysis, it was observed that Akt expression increased 1.092 times, BCL-2 expression decreased approximately 0.3 times, and caspase 3 expression increased 1.2 times. Conclusions: Constituents of Juniperus oxycedrus plant may have apoptotic effects on chronic myeloid leukemia cells.
{"title":"PHYTOCHEMICAL CONSTITUENTS AND ANTILEUKEMIC EFFECTS OF JUNIPERUS OXYCEDRUS EXTRACT","authors":"Berfin Tugba Turak","doi":"10.15407/biotech15.05.064","DOIUrl":"https://doi.org/10.15407/biotech15.05.064","url":null,"abstract":"Aim: Many genetic and environmental factors can be effective in the process of cancerization. Preventing the progression of leukemia may be possible by controlling the pathways involving mechanisms such as apoptosis and autophagy. When the literature is examined, there are studies showing the effects of various types of juniper on various cancer cell lines, including human chronic myeloid leukemia cells, but the signal pathways in which they act are not fully known. In this study, the anticancer effects of Juniperus oxycedrus extract on K-562 human chronic myeloid leukemia cells were investigated. Method: After the cells were treated with the Juniperus oxycedrus extract, cytotoxicity and gene expression analyzes were performed. Changes in the expression of Akt, the member of the PI3K/Akt/mTOR signaling pathway; caspase 3, which is one of the main effective genes in the pathways regulating apoptosis; and the apoptosis suppressor BCL-2 gene, which is an oncogene, were investigated. Results: According to the MTT test results, Juniperus oxycedrus extract showed over approximately 50% cell viability in K-562 cells at all doses. The most appropriate dose of Juniperus oxycedrus fruit extract in this research was determined as 50 µg/ mL considering cell viability. After the gene expression analysis, it was observed that Akt expression increased 1.092 times, BCL-2 expression decreased approximately 0.3 times, and caspase 3 expression increased 1.2 times. Conclusions: Constituents of Juniperus oxycedrus plant may have apoptotic effects on chronic myeloid leukemia cells.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46096685","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 : 2022-10-01DOI: 10.15407/biotech15.05.041
V. A. Didkivskyi
Intravascular thrombosis is one of the main causes of mortality in the working-age population of the world. There are no antithrombotic drugs that act directly on the final stage of thrombosis – fibrin polymerization. However, a new compound of the calix[4]arene series, calix[4]arene C-145, which directly interacts with the fibrin polymerization site ‘A-knob’ thus blocking formation of polymeric fibrin and preventing thrombosis. So, the purpose of this work was to study the calix[4]arene C-145 series as antithrombotic agents in vivo using different animals and types of administration. Materials and methods. Laboratory animals (rats, mice and rabbits) were used for C-145 testing in vivo. Activated partial thromboplastin time and platelet aggregation were measured to determine the anticoagulant action after intravenous or per os administration. Results. Per os way of administration was selected as the optimal one. We showed the substantial prolongation of clotting time in APTT test that was observed starting from the 2nd hour after the per os administration, reached the maximum on 6th hour and eliminated in 24 hours. The effect of C-145 on platelets reached maximum on 4-6 hours and eliminated in 12 hours. Conclusions. C-145 was proven to be prospective antithrombotic drug that can be administered per os. Further investigations must be focused on the study of C-145 pharmacodynamics and metabolism. Such data would allow fast implementation of the tested compound into practice.
{"title":"APPROBATION OF CALIX[4]ARENE AS AN ANTITHROMBOTIC AGENT IN VIVO","authors":"V. A. Didkivskyi","doi":"10.15407/biotech15.05.041","DOIUrl":"https://doi.org/10.15407/biotech15.05.041","url":null,"abstract":"Intravascular thrombosis is one of the main causes of mortality in the working-age population of the world. There are no antithrombotic drugs that act directly on the final stage of thrombosis – fibrin polymerization. However, a new compound of the calix[4]arene series, calix[4]arene C-145, which directly interacts with the fibrin polymerization site ‘A-knob’ thus blocking formation of polymeric fibrin and preventing thrombosis. So, the purpose of this work was to study the calix[4]arene C-145 series as antithrombotic agents in vivo using different animals and types of administration. Materials and methods. Laboratory animals (rats, mice and rabbits) were used for C-145 testing in vivo. Activated partial thromboplastin time and platelet aggregation were measured to determine the anticoagulant action after intravenous or per os administration. Results. Per os way of administration was selected as the optimal one. We showed the substantial prolongation of clotting time in APTT test that was observed starting from the 2nd hour after the per os administration, reached the maximum on 6th hour and eliminated in 24 hours. The effect of C-145 on platelets reached maximum on 4-6 hours and eliminated in 12 hours. Conclusions. C-145 was proven to be prospective antithrombotic drug that can be administered per os. Further investigations must be focused on the study of C-145 pharmacodynamics and metabolism. Such data would allow fast implementation of the tested compound into practice.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48598693","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 : 2022-10-01DOI: 10.15407/biotech15.05.024
D. Pylypenko
Liposomal drug delivery system is an example of the use of nanodrugs in medical practice. Encapsulation of active pharmaceutical ingredients in liposomal nanoparticles allows increasing their bioavailability and efficacy. Aim. The article is devoted to the analysis of the lipid composition of liposomal drugs developed in Ukraine, its influence on the choice of technology and control parameters. Results. The lipid compositions of liposomal drugs developed in Ukraine in recent years were reviewed. The advantages and disadvantages of natural phosphatidylcholine as the main membrane-forming lipid were analyzed. Data on the influence of anionic phospholipids and cholesterol in the liposomal membrane composition on the stability of liposomal nanoparticles and the level of active pharmaceutical ingredient encapsulation were given. The main technological stages of obtaining liposomes with hydrophilic and hydrophobic active pharmaceutical ingredients were considered. The main groups of quality indicators of liposomal dosage forms have been determined. Conclusions. The lipid composition determines the structure and physicochemical properties of the lipid membrane, the mechanism and level of active pharmaceutical ingredient encapsulation, which significantly influences the pharmacological efficacy of liposomal drug delivery systems.
{"title":"INFLUENCE OF THE LIPID COMPOSITION ON THE PROPERTIES, TECHNOLOGY AND QUALITY INDICATORS OF LIPOSOMAL DRUGS","authors":"D. Pylypenko","doi":"10.15407/biotech15.05.024","DOIUrl":"https://doi.org/10.15407/biotech15.05.024","url":null,"abstract":"Liposomal drug delivery system is an example of the use of nanodrugs in medical practice. Encapsulation of active pharmaceutical ingredients in liposomal nanoparticles allows increasing their bioavailability and efficacy. Aim. The article is devoted to the analysis of the lipid composition of liposomal drugs developed in Ukraine, its influence on the choice of technology and control parameters. Results. The lipid compositions of liposomal drugs developed in Ukraine in recent years were reviewed. The advantages and disadvantages of natural phosphatidylcholine as the main membrane-forming lipid were analyzed. Data on the influence of anionic phospholipids and cholesterol in the liposomal membrane composition on the stability of liposomal nanoparticles and the level of active pharmaceutical ingredient encapsulation were given. The main technological stages of obtaining liposomes with hydrophilic and hydrophobic active pharmaceutical ingredients were considered. The main groups of quality indicators of liposomal dosage forms have been determined. Conclusions. The lipid composition determines the structure and physicochemical properties of the lipid membrane, the mechanism and level of active pharmaceutical ingredient encapsulation, which significantly influences the pharmacological efficacy of liposomal drug delivery systems.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47346253","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 : 2022-10-01DOI: 10.15407/biotech15.05.058
S. Magomedov
Acute phase proteins ceruloplasmin, haptoglobin, C-reactive protein (CRP) and procalcitonin are markers that characterize the inflammatory process. C-reactive protein is one of the central components of the acute phase (AF) and is a generally accepted indicator of inflammatory processes. Aim. Determination of the level and efficiency of determination of acute-phase proteins (CRP, haptoglobin, ceruloplasmin), as well as procalcitonin under the conditions of modeling infectious arthritis. Materials and methods. Experimental studies were conducted on 52 white male Wistar rats. A model of infectious arthritis was created for seven days by daily injection of 0.02 ml of S.aureus 108 No. 209 into the knee joint of a rat. The animals were divided into groups - and vivarium control. The following model of drug administration was used for the experimental groups: a single daily injection of 0.02 ml of flosteron into the knee joint for seven days (group II); daily single administration for seven days of 0.02 ml of S.aureus 108 No. 209 (III group); daily one-time alternating (every other day) administration for seven days of 0.02 ml of flosteron and 0.02 ml of S.aureus 108 No. 209 into the knee joint (group IV). The effectiveness of the drugs was observed 3 and 14 days after administration. Results. It was established that the concentration of haptoglobin was significantly increased in the blood serum of rats both after 3 and 14 days in all studied groups of animals compared to the control. The greatest increase relative to the control values was noted 3 days after the seven-time injection of S.aureus 108 #209 into the knee joint. However, after 14 days it was already not so significant and significantly lower (by 85.33%) compared to the measurement after three days. Only in rats after a 14-day alternating (every other day) injection of 0.02 ml of flosteron and 0.02 ml of S.aureus 108 No. 209 into the knee joint was observed a probable increase in the level of haptoglobin by 775.08% (Р<0.05) compared to the control and 77.78% reduced compared to the measurement after three days. The concentration of ceruloplasmin in blood serum increased in all experimental rats during the entire observation period and differed little between 3 and 14 days. The content of C-reactive protein in blood serum increased in all studied groups of rats without exception, which proves its high specificity for detecting inflammatory processes of various severity. The concentration of procalcitonin was most likely to increase by 235.0% 3 days after alternating (every other day) administration of 0.02 ml of flosterone and 0.02 ml of S.aureus 108 No. 209. It was slightly lower by 120.0% under the same conditions experiment after 14 days. This indicator probably increased by 65% 14 days after the 7-time introduction of S.aureus 108 #209. In the rest of the experimental animals, the PCT concentration did not change. Conclusions. The determination of haptoglobin reflects, first of all, the
{"title":"ЕFFICIENCY OF DETERMINATION OF ACUTE PHASE PROTEINS AND PROCALCITONIN UNDER THE CONDITIONS OF SIMULATING INFECTIOUS ARTHRITIS","authors":"S. Magomedov","doi":"10.15407/biotech15.05.058","DOIUrl":"https://doi.org/10.15407/biotech15.05.058","url":null,"abstract":"Acute phase proteins ceruloplasmin, haptoglobin, C-reactive protein (CRP) and procalcitonin are markers that characterize the inflammatory process. C-reactive protein is one of the central components of the acute phase (AF) and is a generally accepted indicator of inflammatory processes. Aim. Determination of the level and efficiency of determination of acute-phase proteins (CRP, haptoglobin, ceruloplasmin), as well as procalcitonin under the conditions of modeling infectious arthritis. Materials and methods. Experimental studies were conducted on 52 white male Wistar rats. A model of infectious arthritis was created for seven days by daily injection of 0.02 ml of S.aureus 108 No. 209 into the knee joint of a rat. The animals were divided into groups - and vivarium control. The following model of drug administration was used for the experimental groups: a single daily injection of 0.02 ml of flosteron into the knee joint for seven days (group II); daily single administration for seven days of 0.02 ml of S.aureus 108 No. 209 (III group); daily one-time alternating (every other day) administration for seven days of 0.02 ml of flosteron and 0.02 ml of S.aureus 108 No. 209 into the knee joint (group IV). The effectiveness of the drugs was observed 3 and 14 days after administration. Results. It was established that the concentration of haptoglobin was significantly increased in the blood serum of rats both after 3 and 14 days in all studied groups of animals compared to the control. The greatest increase relative to the control values was noted 3 days after the seven-time injection of S.aureus 108 #209 into the knee joint. However, after 14 days it was already not so significant and significantly lower (by 85.33%) compared to the measurement after three days. Only in rats after a 14-day alternating (every other day) injection of 0.02 ml of flosteron and 0.02 ml of S.aureus 108 No. 209 into the knee joint was observed a probable increase in the level of haptoglobin by 775.08% (Р<0.05) compared to the control and 77.78% reduced compared to the measurement after three days. The concentration of ceruloplasmin in blood serum increased in all experimental rats during the entire observation period and differed little between 3 and 14 days. The content of C-reactive protein in blood serum increased in all studied groups of rats without exception, which proves its high specificity for detecting inflammatory processes of various severity. The concentration of procalcitonin was most likely to increase by 235.0% 3 days after alternating (every other day) administration of 0.02 ml of flosterone and 0.02 ml of S.aureus 108 No. 209. It was slightly lower by 120.0% under the same conditions experiment after 14 days. This indicator probably increased by 65% 14 days after the 7-time introduction of S.aureus 108 #209. In the rest of the experimental animals, the PCT concentration did not change. Conclusions. The determination of haptoglobin reflects, first of all, the ","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43396492","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 : 2022-10-01DOI: 10.15407/biotech15.05.047
Onosetale E. Aigbomian
Studies have shown that pain relieving medications may be neuroprotective. Ocimum gratissimum Linn. that is widely used in traditional medicine for debility and many other illnesses neuropharmacologically related has not been fully explored. Aim. This study was designed to investigate the safety of intake, neurobehavioral and analgesic effects of the Essential Oil of Ocimum gratissimum Linn leaves (EOOG) in mice. Methods. Acute toxicity of EOOG was determined following standard method while the neurobehavioural properties were assessed using the open field for Novelty-Induced Rearing (NIR), Novelty-Induced Grooming (NIG) and locomotor activity in mice. The hole board apparatus was used for the frequency of head dips. The Y-maze was used for short- working memory. Mechanistic studies were conducted with Atropine (muscarinic blocker, 0.5 mg/kg), Propanolol (non-selective ß-adrenoceptor blocker, 0.2 mg/kg), Haloperidol (dopamine receptor blocker, 0.2 mg/kg), Cyproheptadine (Serotoninergic antagonist, 0.5 mg/kg) and Yohimbine (ά-2 adrenergic blocker, 1 mg/kg). The analgesic activity of Ocimum gratissimum was investigated using acetic acid writhing test and thermally-induced pain. Results. The median lethal dose (LD50) of Ocimum gratissimum was 2449 mg/kg. The EOOG significantly reduced novelty-induced behaviour in a dose-dependent manner. The exploratory activity of animals treated with the EOOG was observed to decrease non-dose dependently with the highest dose (40 mg/kg) showing no activity on the hole board apparatus. The EOOG produced a significant reduction in locomotor activity in all the doses in a non-dose dependent manner but at the lowest dose. In the Y-maze, EOOG did not produce any significant effect on working memory as the percentage alternation produced was not significantly different from the control. The EOOG in hot plate analgesic assay showed increased reaction time suggesting central nervous system analgesic property. Conclusions. The results of the investigation showed that EOOG might possess sedative properties due to its ability to inhibit NIR and NIG, head dips, and locomotor activity. Furthermore, the inhibition of nociception marked in this research advocates antinociceptive activity which might be through the peripheral or central opioid receptor..
{"title":"NEUROBEHAVIOURAL AND ANALGESIC EFFECT OF Ocimum gratissimum LINN. LEAVES ESSENTIAL OIL IN Wistar albino MICE","authors":"Onosetale E. Aigbomian","doi":"10.15407/biotech15.05.047","DOIUrl":"https://doi.org/10.15407/biotech15.05.047","url":null,"abstract":"Studies have shown that pain relieving medications may be neuroprotective. Ocimum gratissimum Linn. that is widely used in traditional medicine for debility and many other illnesses neuropharmacologically related has not been fully explored. Aim. This study was designed to investigate the safety of intake, neurobehavioral and analgesic effects of the Essential Oil of Ocimum gratissimum Linn leaves (EOOG) in mice. Methods. Acute toxicity of EOOG was determined following standard method while the neurobehavioural properties were assessed using the open field for Novelty-Induced Rearing (NIR), Novelty-Induced Grooming (NIG) and locomotor activity in mice. The hole board apparatus was used for the frequency of head dips. The Y-maze was used for short- working memory. Mechanistic studies were conducted with Atropine (muscarinic blocker, 0.5 mg/kg), Propanolol (non-selective ß-adrenoceptor blocker, 0.2 mg/kg), Haloperidol (dopamine receptor blocker, 0.2 mg/kg), Cyproheptadine (Serotoninergic antagonist, 0.5 mg/kg) and Yohimbine (ά-2 adrenergic blocker, 1 mg/kg). The analgesic activity of Ocimum gratissimum was investigated using acetic acid writhing test and thermally-induced pain. Results. The median lethal dose (LD50) of Ocimum gratissimum was 2449 mg/kg. The EOOG significantly reduced novelty-induced behaviour in a dose-dependent manner. The exploratory activity of animals treated with the EOOG was observed to decrease non-dose dependently with the highest dose (40 mg/kg) showing no activity on the hole board apparatus. The EOOG produced a significant reduction in locomotor activity in all the doses in a non-dose dependent manner but at the lowest dose. In the Y-maze, EOOG did not produce any significant effect on working memory as the percentage alternation produced was not significantly different from the control. The EOOG in hot plate analgesic assay showed increased reaction time suggesting central nervous system analgesic property. Conclusions. The results of the investigation showed that EOOG might possess sedative properties due to its ability to inhibit NIR and NIG, head dips, and locomotor activity. Furthermore, the inhibition of nociception marked in this research advocates antinociceptive activity which might be through the peripheral or central opioid receptor..","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47524170","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 : 2022-10-01DOI: 10.15407/biotech15.05.031
A. Tykhomyrov
Lactoferrin is a ubiquitous and multifunctional protein, which has antimicrobial and immunomodulatory activities. Lactoferrin plays an important role in the maintenance of ocular health. The aim of the study was to produce polyclonal antibodies against human lactoferrin in order to apply them in evaluation of lactoferrin levels in tear fluid collected from healthy eye and after corneal injury. Materials and methods. Affine chromatography on Protein A-sepharose was applied in order to isolate immunoglobulin G (IgG) fraction from the blood serum of lactoferrin-immunized rabbits. Each step of protein purification was monitored by denaturing gel electrophoresis (SDS-PAGE). Target antigen recognition by produced antibodies was established by western blot analysis with the use of diluted IgG fraction. Lactoferrin levels in the tear fluids collected from healthy individuals (n = 4) and patients with non-penetrating corneal injures (n = 6) were determined immunochemically with the use of purified antibodies. The results of western blot of lactoferrin levels in the tear fluids of healthy individuals and patients with corneal wounds were analysed using Mann-Whitney U-test. The difference between group mean values was considered significant at P<0.05. Results. Using affine chromatography on Protein A-sepharose, antibodies against human lactoferrin were purified as IgG fraction from blood serum of lactoferrin-immunized rabbits. Western blot analysis showed that obtained antibodies recognize the antigen as a 75-kDa band, which corresponds to the intact human lactoferrin polypeptide. The same major polypeptide band was visualized by western blot with enhanced chemiluminescence detection in the tear fluid samples. Densitometry analysis of 75-kDa lactoferrin band showed 3.2-fold decrease in lactoferrin level in the tear fluid samples obtained from patients with non-penetrating corneal traumas as compared with samples collected from healthy persons (P<0.05). Besides, tear fluid of patients with injured corneas contained large amounts of truncated lactoferrin immunoreactive polypeptides as well as high molecular weight bands, which could correspond to lactoferrin complexes with other proteins occurring during inflammation. Conclusions. According to our data, obtained anti-lactoferrin antibodies can be used as a valuable tool for development of advanced tests and procedures for diagnostics of eye diseases associated with the corneal lesions. Reduced lactoferrin concentration might represent a potential prognostic biomarker for diagnosis of ocular diseases including non-penetrating corneal injuries in a simple and non-invasive way.
{"title":"PRODUCTION OF ANTI-LACTOFERRIN ANTIBODIES AND THEIR APPLICATION IN ANALYSIS OF THE TEAR FLUID IN HEALTH AND CORNEAL INJURIES","authors":"A. Tykhomyrov","doi":"10.15407/biotech15.05.031","DOIUrl":"https://doi.org/10.15407/biotech15.05.031","url":null,"abstract":"Lactoferrin is a ubiquitous and multifunctional protein, which has antimicrobial and immunomodulatory activities. Lactoferrin plays an important role in the maintenance of ocular health. The aim of the study was to produce polyclonal antibodies against human lactoferrin in order to apply them in evaluation of lactoferrin levels in tear fluid collected from healthy eye and after corneal injury. Materials and methods. Affine chromatography on Protein A-sepharose was applied in order to isolate immunoglobulin G (IgG) fraction from the blood serum of lactoferrin-immunized rabbits. Each step of protein purification was monitored by denaturing gel electrophoresis (SDS-PAGE). Target antigen recognition by produced antibodies was established by western blot analysis with the use of diluted IgG fraction. Lactoferrin levels in the tear fluids collected from healthy individuals (n = 4) and patients with non-penetrating corneal injures (n = 6) were determined immunochemically with the use of purified antibodies. The results of western blot of lactoferrin levels in the tear fluids of healthy individuals and patients with corneal wounds were analysed using Mann-Whitney U-test. The difference between group mean values was considered significant at P<0.05. Results. Using affine chromatography on Protein A-sepharose, antibodies against human lactoferrin were purified as IgG fraction from blood serum of lactoferrin-immunized rabbits. Western blot analysis showed that obtained antibodies recognize the antigen as a 75-kDa band, which corresponds to the intact human lactoferrin polypeptide. The same major polypeptide band was visualized by western blot with enhanced chemiluminescence detection in the tear fluid samples. Densitometry analysis of 75-kDa lactoferrin band showed 3.2-fold decrease in lactoferrin level in the tear fluid samples obtained from patients with non-penetrating corneal traumas as compared with samples collected from healthy persons (P<0.05). Besides, tear fluid of patients with injured corneas contained large amounts of truncated lactoferrin immunoreactive polypeptides as well as high molecular weight bands, which could correspond to lactoferrin complexes with other proteins occurring during inflammation. Conclusions. According to our data, obtained anti-lactoferrin antibodies can be used as a valuable tool for development of advanced tests and procedures for diagnostics of eye diseases associated with the corneal lesions. Reduced lactoferrin concentration might represent a potential prognostic biomarker for diagnosis of ocular diseases including non-penetrating corneal injuries in a simple and non-invasive way.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45733519","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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