Pub Date : 2022-08-31DOI: 10.15407/biotech15.04.022
Y. Nesterenko
Aim. To investigate electrolytic aggregation of different nano-objects in solutions with quantum dots (QDs) and Au nanoparticles (NPs) modified by oligonucleotides as well as the effect of aggregates on the photoluminescence (PL) of QDs. Methods. Au NPs and AgInS2/ZnS QDs were modified by oligonucleotides. Two types of QDs that differ in size and stabilizing ligand were used. PL and optical absorption of nano-objects in water and SSC buffer solutions were studied. Results. The transfer of modified by oligonucleotides QDs from water to a buffer solution and the addition of Au NP modified by oligonucleotides to the solution caused quenching of the QD PL intensity. The PL quenching was observed for the QDs of two types and increased during the incubation of solutions, but didn’t depend on its multiplicity. An aggregation of Au-DP occurred only in buffer solutions with QDs of one type and increased with multiplicity of the buffer solution. Conclusion. It is found that the electrolytic aggregation of Au NPs modified by oligonucleotides in buffer solutions with QDs depends on the QD type and didn’t affect the quenching of the PL intensity of the QDs.
{"title":"ELECTROLYTIC AGGREGATION IN SOLUTIONS WITH QUANTUM DOTS AND GOLD NANOPARTICLES MODIFIED WITH OLIGONUCLEOTIDES","authors":"Y. Nesterenko","doi":"10.15407/biotech15.04.022","DOIUrl":"https://doi.org/10.15407/biotech15.04.022","url":null,"abstract":"Aim. To investigate electrolytic aggregation of different nano-objects in solutions with quantum dots (QDs) and Au nanoparticles (NPs) modified by oligonucleotides as well as the effect of aggregates on the photoluminescence (PL) of QDs. Methods. Au NPs and AgInS2/ZnS QDs were modified by oligonucleotides. Two types of QDs that differ in size and stabilizing ligand were used. PL and optical absorption of nano-objects in water and SSC buffer solutions were studied. Results. The transfer of modified by oligonucleotides QDs from water to a buffer solution and the addition of Au NP modified by oligonucleotides to the solution caused quenching of the QD PL intensity. The PL quenching was observed for the QDs of two types and increased during the incubation of solutions, but didn’t depend on its multiplicity. An aggregation of Au-DP occurred only in buffer solutions with QDs of one type and increased with multiplicity of the buffer solution. Conclusion. It is found that the electrolytic aggregation of Au NPs modified by oligonucleotides in buffer solutions with QDs depends on the QD type and didn’t affect the quenching of the PL intensity of the QDs.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45456437","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 : 2022-08-31DOI: 10.15407/biotech15.04.030
K. Rebenko
COVID-19 disease, an acute respiratory infection caused by the SARS-CoV-2 virus, manifests itself in various severity forms - mild, moderate and severe, caused by the reactions of the patient's immune response. Aim. To evaluate the serum levels of immunoglobulins G, M, and A and the number of circulating monocytes of different phenotypes in female patients with the abovementioned forms of COVID-19 severity. Methods. Blood samples of 53 women with SARS-CoV-2 infection were studied. Flow cytofluorimetry was used to estimate monocyte subpopulations by the expression of CD14 and CD16. Concentrations of IgM, IgG, and IgA in the serum were determined in radial immunodiffusion test according to Mancini. Results. The relative number of non-classical monocytes with CD14+-CD16++ phenotype was significantly decreased in the blood of COVID-19 patients from all 3 clinical severity groups, while changes in the number of classical and intermediate monocytes were insignificant. The levels of IgA in COVID-19 patients significantly decreased after recovery as compared to the acute phase of the infection. Conclusion. The results emphasize the importance of monocyte subpopulation analysis in COVID-19 diagnosis and indicate dynamic changes in IgA levels depending on disease severity. The research data may help in the development of new diagnosis methods and therapy for SARS-CoV-2 infection.
{"title":"ІMMUNOGLOBULIN ISOTYPES AND BLOOD MONOCYTE SUBPOPULATIONS IN COVID-19 FEMALE PATIENTS WITH DIFFERENT DISEASE SEVERITY","authors":"K. Rebenko","doi":"10.15407/biotech15.04.030","DOIUrl":"https://doi.org/10.15407/biotech15.04.030","url":null,"abstract":"COVID-19 disease, an acute respiratory infection caused by the SARS-CoV-2 virus, manifests itself in various severity forms - mild, moderate and severe, caused by the reactions of the patient's immune response. Aim. To evaluate the serum levels of immunoglobulins G, M, and A and the number of circulating monocytes of different phenotypes in female patients with the abovementioned forms of COVID-19 severity. Methods. Blood samples of 53 women with SARS-CoV-2 infection were studied. Flow cytofluorimetry was used to estimate monocyte subpopulations by the expression of CD14 and CD16. Concentrations of IgM, IgG, and IgA in the serum were determined in radial immunodiffusion test according to Mancini. Results. The relative number of non-classical monocytes with CD14+-CD16++ phenotype was significantly decreased in the blood of COVID-19 patients from all 3 clinical severity groups, while changes in the number of classical and intermediate monocytes were insignificant. The levels of IgA in COVID-19 patients significantly decreased after recovery as compared to the acute phase of the infection. Conclusion. The results emphasize the importance of monocyte subpopulation analysis in COVID-19 diagnosis and indicate dynamic changes in IgA levels depending on disease severity. The research data may help in the development of new diagnosis methods and therapy for SARS-CoV-2 infection.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41594375","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 : 2022-08-31DOI: 10.15407/biotech15.04.044
Y. Susak
The purpose of this study was comparative assessment of the frequency of bacterial colonization of the bile in patients with hilar malignant biliary obstruction after the palliative biliary decompression using different methodological approaches. Methods. 50 patients with proximal mechanical jaundice of tumor origin aged of ~ 62 years (25 males and 25 females), who were on steady-state treatment in Main military clinical hospital (Kyiv, Ukraine) were recruited in this prospective study. All patients underwent cholangiostomy using percutaneous transhepatic (PTBD) and external-internal suprapapillary (EISBD) approaches. Bile specimens were taken right after the biliary drainage. Identification of bacterial isolates was performed using standard cultural and biochemical methods. Results. The incidence of cholangitis was almost twice lower in EISBD group (n=26) than in PTBD group (n=24): 25.6% vs 49.1%. The rates of bacteriobilia did not differ significantly in patients from different groups: 23.1% in EISBD group and 25.0% in PTBD group. However, the frequency of biliary bacterial colonization coupled with cholangitis was also 2 times lower in EISBD group in comparison with patients underwent PTBD: 7.7% vs 16.7%. Escherichia coli predominated in bile specimens from patients with bacteriobilia associated with cholangitis in both groups. Conclusions. The use of EISBD for palliative biliary decompression in patients with proximal mechanical jaundice of tumor origin is associated with lower risk of bacterial colonization of the bile as compared to PTBD approach, and as a result with less risk of the development of infectious complications
{"title":"THE IMPACT OF BILIARY DRAINAGE MODE ON BACTERIOBILIA OCCURRENCE IN PATIENTS WITH HILAR MALIGNANT OBSTRUCTION","authors":"Y. Susak","doi":"10.15407/biotech15.04.044","DOIUrl":"https://doi.org/10.15407/biotech15.04.044","url":null,"abstract":"The purpose of this study was comparative assessment of the frequency of bacterial colonization of the bile in patients with hilar malignant biliary obstruction after the palliative biliary decompression using different methodological approaches. Methods. 50 patients with proximal mechanical jaundice of tumor origin aged of ~ 62 years (25 males and 25 females), who were on steady-state treatment in Main military clinical hospital (Kyiv, Ukraine) were recruited in this prospective study. All patients underwent cholangiostomy using percutaneous transhepatic (PTBD) and external-internal suprapapillary (EISBD) approaches. Bile specimens were taken right after the biliary drainage. Identification of bacterial isolates was performed using standard cultural and biochemical methods. Results. The incidence of cholangitis was almost twice lower in EISBD group (n=26) than in PTBD group (n=24): 25.6% vs 49.1%. The rates of bacteriobilia did not differ significantly in patients from different groups: 23.1% in EISBD group and 25.0% in PTBD group. However, the frequency of biliary bacterial colonization coupled with cholangitis was also 2 times lower in EISBD group in comparison with patients underwent PTBD: 7.7% vs 16.7%. Escherichia coli predominated in bile specimens from patients with bacteriobilia associated with cholangitis in both groups. Conclusions. The use of EISBD for palliative biliary decompression in patients with proximal mechanical jaundice of tumor origin is associated with lower risk of bacterial colonization of the bile as compared to PTBD approach, and as a result with less risk of the development of infectious complications","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47764201","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 : 2022-08-31DOI: 10.15407/biotech15.04.034
D. A. Shustyk
Aim. To examine the effect of endophytic bacteria Paenibacillus polymyxa P 6.3, which was isolated from grains of winter wheat variety Podolyanka, on the germination of wheat grains after the exposure Pseudomonas syringae. Methods. Growth-promoting and biocontrol activity of P. polymyxa P 6.3 were examined using roll method. Standardized wheat grains were soaked in a suspension of 24 h culture of P. polymyxa P 6.3 for 12 h, control grains were soaked in sterile distilled H2O. After soaking, both pre-treated and control grains (of 25 pcs) were put into rolls. In three days, half of both pre-treated and control grains were exposed to phytopathogen P. sуringae, and germination was continued. The lengths of coleoptile and main root were measured on the 7th day of the experiment. Results were expressed as M ± m. Differences were considered significant at P ≤ 0.05. Results. Treatment wheat grains with P. polymyxa P 6.3 resulted in increased growth of coleoptile and main root in all three varieties. Most prominent effect was registered in Favorytka variety. After the exposure to phytopathogenic pseudomonads, slowing down of the growth of coleoptile and main root occurred in all wheat varieties. Highest susceptibility to P. syringae pathogenic effect was registered in Holikovska variety. Pre-treatment of wheat grains with endophytic bacteria abrogated growth-inhibiting effects of P.syringae. Conclusion. Endophytic bacteria P. polymyxa P 6.3 exerts a growth-stimulating effect on wheat germination and a protective effect against P. syringae. The plant growth promoting potential and antagonistic activity make strain P 6.3 a promising biocontrol agent and growth stimulator as a biofertilizer.
{"title":"GRAIN-RESIDING ENDOPHYTIC BACTERIUM Paenibacillus polymyxa P 6.3 POSSESSES GROWTH-PROMOTING ACTIVITY AND PROTECT WHEAT GRAIN FROM PATHOGENIC EFFECT OF Pseudomonas syringae","authors":"D. A. Shustyk","doi":"10.15407/biotech15.04.034","DOIUrl":"https://doi.org/10.15407/biotech15.04.034","url":null,"abstract":"Aim. To examine the effect of endophytic bacteria Paenibacillus polymyxa P 6.3, which was isolated from grains of winter wheat variety Podolyanka, on the germination of wheat grains after the exposure Pseudomonas syringae. Methods. Growth-promoting and biocontrol activity of P. polymyxa P 6.3 were examined using roll method. Standardized wheat grains were soaked in a suspension of 24 h culture of P. polymyxa P 6.3 for 12 h, control grains were soaked in sterile distilled H2O. After soaking, both pre-treated and control grains (of 25 pcs) were put into rolls. In three days, half of both pre-treated and control grains were exposed to phytopathogen P. sуringae, and germination was continued. The lengths of coleoptile and main root were measured on the 7th day of the experiment. Results were expressed as M ± m. Differences were considered significant at P ≤ 0.05. Results. Treatment wheat grains with P. polymyxa P 6.3 resulted in increased growth of coleoptile and main root in all three varieties. Most prominent effect was registered in Favorytka variety. After the exposure to phytopathogenic pseudomonads, slowing down of the growth of coleoptile and main root occurred in all wheat varieties. Highest susceptibility to P. syringae pathogenic effect was registered in Holikovska variety. Pre-treatment of wheat grains with endophytic bacteria abrogated growth-inhibiting effects of P.syringae. Conclusion. Endophytic bacteria P. polymyxa P 6.3 exerts a growth-stimulating effect on wheat germination and a protective effect against P. syringae. The plant growth promoting potential and antagonistic activity make strain P 6.3 a promising biocontrol agent and growth stimulator as a biofertilizer.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44905736","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 : 2022-08-31DOI: 10.15407/biotech15.04.047
T. V. Symchych
Cancer cells upregulate surface expression of N-glycolyl-neuraminic acid (Neu5Gc). At Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology (IEPOR) of the National Academy of Science of Ukraine, B. subtilis IMV В-7724 lectin specific for Neu5Gc was obtained. Aim. The scope of the research was to study in vitro B. subtilis IMV В-7724 lectin activity towards malignant and normal cells. Materials and methods. Cytotoxic and mitogenic activities was studied by, respectively, MTT-assay and in vitro lymphocytes proliferation assay. Results. The lectin possesses cytotoxic activity towards human (А549, HL60) and murine (Ehrlich carcinoma, L1210) cancer cell lines. The most sensitive were L1210 and Ehrlich carcinoma cell lines. IC50 was 0.16 mg/ml in both cases. The lectin was less cytotoxic to murine peritoneal macrophages, lymphocytes and thymocytes: IC50 was 0.47, 2.02 and 3.49 mg/ml respectively. In a dose of 25 µg/ml the lectin induced lymphocytes proliferation. Conclusion. Depending on the target cells type and applied dose, B. subtilis IMV B-7724 lectin shows cytotoxic or mitogenic activities. Both of lectin’s activities can be applied in cancer treatment and thus deserve further investigation
{"title":"EFFECTS OF B. Subtilis IMV В-7724 LECTIN ON MALIGNANT AND NORMAL CELLS in vitro","authors":"T. V. Symchych","doi":"10.15407/biotech15.04.047","DOIUrl":"https://doi.org/10.15407/biotech15.04.047","url":null,"abstract":"Cancer cells upregulate surface expression of N-glycolyl-neuraminic acid (Neu5Gc). At Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology (IEPOR) of the National Academy of Science of Ukraine, B. subtilis IMV В-7724 lectin specific for Neu5Gc was obtained. Aim. The scope of the research was to study in vitro B. subtilis IMV В-7724 lectin activity towards malignant and normal cells. Materials and methods. Cytotoxic and mitogenic activities was studied by, respectively, MTT-assay and in vitro lymphocytes proliferation assay. Results. The lectin possesses cytotoxic activity towards human (А549, HL60) and murine (Ehrlich carcinoma, L1210) cancer cell lines. The most sensitive were L1210 and Ehrlich carcinoma cell lines. IC50 was 0.16 mg/ml in both cases. The lectin was less cytotoxic to murine peritoneal macrophages, lymphocytes and thymocytes: IC50 was 0.47, 2.02 and 3.49 mg/ml respectively. In a dose of 25 µg/ml the lectin induced lymphocytes proliferation. Conclusion. Depending on the target cells type and applied dose, B. subtilis IMV B-7724 lectin shows cytotoxic or mitogenic activities. Both of lectin’s activities can be applied in cancer treatment and thus deserve further investigation","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46565930","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 : 2022-06-30DOI: 10.15407/biotech15.03.029
R. Marunych
Chelidonium majus is a well-known source of biologically active compounds. Most of them are alkaloids, which are used in researches and for tradition medicine. In this study, we explored the influence of C. majus crude total extract onto blood coagulation system in vitro, primary and secondary hemostasis. Aim. To study influence of C. majus extract onto blood coagulation process. Methods. Turbidimetry of blood plasma, APTT test with chromogenic substrates, and platelet aggregation were used in the work. Results. We demonstrated moderate stimulating effect of the extract on platelets (the rate of platelet aggregation increased up to 10%, followed by disaggregation). Extract also increased the rate of platelet aggregation stimulated by 12.5 mcM of ADP. We observed the increase in the plasma clotting time in the presence of the extract, that corresponded to the 274, 411, 685, 1370 mcg of dry extract, from 70 s in control to 80, 90, 170 and 180 s, respectively, in turbidimetry test of plasma stimulated by APTT-reagent. However, APTT test with the chromogenic substrate of thrombin (S2238) did not show significant influence of this plant’s extract on thrombin activity. Conclusions. Thus, we can conclude that anticoagulant activity of Chelidonium majus extract corresponded to the direct inhibition of fibrin polymerization.
{"title":"Chelidonium majus WATER EXTRACT INITIATES PLATELET AGGREGATION AND INHIBITS FIBRIN POLYMERIZATION IN BLOOD PLASMA","authors":"R. Marunych","doi":"10.15407/biotech15.03.029","DOIUrl":"https://doi.org/10.15407/biotech15.03.029","url":null,"abstract":"Chelidonium majus is a well-known source of biologically active compounds. Most of them are alkaloids, which are used in researches and for tradition medicine. In this study, we explored the influence of C. majus crude total extract onto blood coagulation system in vitro, primary and secondary hemostasis. Aim. To study influence of C. majus extract onto blood coagulation process. Methods. Turbidimetry of blood plasma, APTT test with chromogenic substrates, and platelet aggregation were used in the work. Results. We demonstrated moderate stimulating effect of the extract on platelets (the rate of platelet aggregation increased up to 10%, followed by disaggregation). Extract also increased the rate of platelet aggregation stimulated by 12.5 mcM of ADP. We observed the increase in the plasma clotting time in the presence of the extract, that corresponded to the 274, 411, 685, 1370 mcg of dry extract, from 70 s in control to 80, 90, 170 and 180 s, respectively, in turbidimetry test of plasma stimulated by APTT-reagent. However, APTT test with the chromogenic substrate of thrombin (S2238) did not show significant influence of this plant’s extract on thrombin activity. Conclusions. Thus, we can conclude that anticoagulant activity of Chelidonium majus extract corresponded to the direct inhibition of fibrin polymerization.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42767893","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 : 2022-06-30DOI: 10.15407/biotech15.03.025
P. Y. Tsap
Aim. Study of the role of the complex between the αC region and the BβN domain in the initial stages of fibrin polymerization has been investigated. Materials and Methods. Method of turbidimetry to study the influence of fibrinogen fragments D and DD on the polymerization and methods of isolation, purification, fragmentation for fibrinogen, monomer and cross-linked fibrin, fibrinogen X-fragment, Glu -plasminogen were used. Results. It was shown that fragment DD completely inhibited polymerization process in all the systems examined (“Fg + Thr”, “Fg + Anc H”, “X + Thr”, “X + Anc H”). Fragment D inhibited fibrin polymerization at all stages in the system “Fg + Thr”, but in the system “Fg + Anc H” it almost did not influence fibrin polymerization. In the both systems “X + Thr” and “X + Anc H” fragment D weakly inhibited the self-assembly of fibrin molecules into protofibrils, but accelerated the process of lateral association in the second system. Conclusions. The data obtained indicated that the complex between the αC region and the BβN domain of fibrin desA, on the initial stage of polymerization supported the rate of self-assembling and lateral association of fibrin desA protofibrils, protecting the oligomers against the depolymerizing influence of fibrinogen.
{"title":"INFLUENCE OF FIBRIN D AND DD FRAGMENTS ON FIBRINOGEN AND FIBRINOGEN FRAGMENT X POLYMERIZATION INITIATED BY THROMBIN OR ANCISTRON","authors":"P. Y. Tsap","doi":"10.15407/biotech15.03.025","DOIUrl":"https://doi.org/10.15407/biotech15.03.025","url":null,"abstract":"Aim. Study of the role of the complex between the αC region and the BβN domain in the initial stages of fibrin polymerization has been investigated. Materials and Methods. Method of turbidimetry to study the influence of fibrinogen fragments D and DD on the polymerization and methods of isolation, purification, fragmentation for fibrinogen, monomer and cross-linked fibrin, fibrinogen X-fragment, Glu -plasminogen were used. Results. It was shown that fragment DD completely inhibited polymerization process in all the systems examined (“Fg + Thr”, “Fg + Anc H”, “X + Thr”, “X + Anc H”). Fragment D inhibited fibrin polymerization at all stages in the system “Fg + Thr”, but in the system “Fg + Anc H” it almost did not influence fibrin polymerization. In the both systems “X + Thr” and “X + Anc H” fragment D weakly inhibited the self-assembly of fibrin molecules into protofibrils, but accelerated the process of lateral association in the second system. Conclusions. The data obtained indicated that the complex between the αC region and the BβN domain of fibrin desA, on the initial stage of polymerization supported the rate of self-assembling and lateral association of fibrin desA protofibrils, protecting the oligomers against the depolymerizing influence of fibrinogen.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44197066","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 : 2022-06-30DOI: 10.15407/biotech15.03.005
N. Hrytseva
Members of Ralstonia solanacearum species complex (RSSC) are causal agents of vascular wilt disease in more than 450 crop species, including solanaceous plants such as potatoes, tomatoes, bell pepper, eggplant, etc. These phytopathogens cause serious yield loss mostly in solanaceous crops which are grown in tropical, subtropical, and temperate regions of the world. Yield losses comprise 80%–100% in potato, up to 91% for tomato, 10%–30% in tobacco, 33%–90% in banana, and reduce crop productivity and yield. PCR-methods are specific, sensitive and cost-effective approaches for the detection and identification of RSSC members. The objective of this study was to compare specificity of routinely used primer mix for PCR RSSC detection with the newly developed pairs of species-specific primers for ease of use diagnostics in a laboratory. Materials and Methods. The conserved genomic regions of the 16S rRNA sequences of R. solanacearum, R. pseudosolanacearum, and R. syzygii were selected for the design of primers for this study. Newly created primer species specificity was tested in PCR using the DNA of the two targets and 13 non-target strains of bacteria. Results. Three pairs of newly created primers Rs-28(F)/Rs-193(R), Rs-28(F)/OLI-160(R), Rs28(F)/OLI248(R) produced single specific fragments for bacterial strains of Ralstonia solanacearum: 166 bp, 132 bp, and 220 bp. products respectively. No PCR products were obtained during amplification with the negative control or non-target DNA templates from other bacterial species. Conclusion. Designed primers can be used for the development of PCR system for the qualitative and quantitative detection of RSSC members.
{"title":"DEVELOPMENT OF SPECIFIC PRIMERS FOR 16S rRNA GENE ANALYSIS IN THE DETECTION OF Ralstonia solanacearum SPECIES COMPLEX","authors":"N. Hrytseva","doi":"10.15407/biotech15.03.005","DOIUrl":"https://doi.org/10.15407/biotech15.03.005","url":null,"abstract":"Members of Ralstonia solanacearum species complex (RSSC) are causal agents of vascular wilt disease in more than 450 crop species, including solanaceous plants such as potatoes, tomatoes, bell pepper, eggplant, etc. These phytopathogens cause serious yield loss mostly in solanaceous crops which are grown in tropical, subtropical, and temperate regions of the world. Yield losses comprise 80%–100% in potato, up to 91% for tomato, 10%–30% in tobacco, 33%–90% in banana, and reduce crop productivity and yield. PCR-methods are specific, sensitive and cost-effective approaches for the detection and identification of RSSC members. The objective of this study was to compare specificity of routinely used primer mix for PCR RSSC detection with the newly developed pairs of species-specific primers for ease of use diagnostics in a laboratory. Materials and Methods. The conserved genomic regions of the 16S rRNA sequences of R. solanacearum, R. pseudosolanacearum, and R. syzygii were selected for the design of primers for this study. Newly created primer species specificity was tested in PCR using the DNA of the two targets and 13 non-target strains of bacteria. Results. Three pairs of newly created primers Rs-28(F)/Rs-193(R), Rs-28(F)/OLI-160(R), Rs28(F)/OLI248(R) produced single specific fragments for bacterial strains of Ralstonia solanacearum: 166 bp, 132 bp, and 220 bp. products respectively. No PCR products were obtained during amplification with the negative control or non-target DNA templates from other bacterial species. Conclusion. Designed primers can be used for the development of PCR system for the qualitative and quantitative detection of RSSC members.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45730405","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 : 2022-06-30DOI: 10.15407/biotech15.03.052
D. S. Korolova
Aim. Treatment by indirect anticoagulants (vitamin K antagonists) requires a personalized approach for controlling the overall level of prothrombin and the accumulation of its decarboxylated forms. The purpose of this work was to optimize the method for monitoring of the therapy with indirect anticoagulants. Methods. An analysis was performed of 41 blood plasma samples from patients with cardiovascula pathologies. Activated partial thromboplastin time (APTT), prothrombin time, ecamulin time, statistical data analysis (“Statistica 7”) have been used. Results. APTT test allowed identifying the individual sensitivity of patients to indirect anticoagulants. In particular, 20% of patients showed a decrease in the total level of prothrombin, which, together with the accumulation of decarboxylated forms, leads to a risk of bleeding. Individual insensitivity to the action of vitamin K antagonists was determined in 11% of patients. Conclusion. To control the efficacy of indirect anticoagulants therapy, we developed test in which ecamulin (protease from the venom of Echis multisquamatis) was used as a prothrombin activator, which can activate not only functionally active prothrombin, but also its decarboxylated forms. Use of ecamulin simultaneously with thromboplastin allows determining in the blood plasma the content of not only functionally active prothrombin, but also the total level of prothrombin, which makes it possible to control the accumulation of decarboxylated prothrombin.
{"title":"OPTIMIZATION OF THE EVALUATION METHOD OF THE PERFORMANCE OF THERAPY USING INDIRECT ACTION ANTICOAGULANTS","authors":"D. S. Korolova","doi":"10.15407/biotech15.03.052","DOIUrl":"https://doi.org/10.15407/biotech15.03.052","url":null,"abstract":"Aim. Treatment by indirect anticoagulants (vitamin K antagonists) requires a personalized approach for controlling the overall level of prothrombin and the accumulation of its decarboxylated forms. The purpose of this work was to optimize the method for monitoring of the therapy with indirect anticoagulants. Methods. An analysis was performed of 41 blood plasma samples from patients with cardiovascula pathologies. Activated partial thromboplastin time (APTT), prothrombin time, ecamulin time, statistical data analysis (“Statistica 7”) have been used. Results. APTT test allowed identifying the individual sensitivity of patients to indirect anticoagulants. In particular, 20% of patients showed a decrease in the total level of prothrombin, which, together with the accumulation of decarboxylated forms, leads to a risk of bleeding. Individual insensitivity to the action of vitamin K antagonists was determined in 11% of patients. Conclusion. To control the efficacy of indirect anticoagulants therapy, we developed test in which ecamulin (protease from the venom of Echis multisquamatis) was used as a prothrombin activator, which can activate not only functionally active prothrombin, but also its decarboxylated forms. Use of ecamulin simultaneously with thromboplastin allows determining in the blood plasma the content of not only functionally active prothrombin, but also the total level of prothrombin, which makes it possible to control the accumulation of decarboxylated prothrombin.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46483185","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 : 2022-06-30DOI: 10.15407/biotech15.03.013
Т. А. Bohdanovych
Aim. To compare Artemisia tilesii “hairy” root lines with different transferred genes in terms of the relationship between the total content of flavonoids, the levels of antioxidant activity (AOA) and reducing power (RP), as well as the activity of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), rolB and rolC genes. Methods. We compared the root lines Nos. 10 and 16, obtained by transformation with the wild Agrobacterium rhizogenes strain A4, lines Nos. 2 and 4, obtained using A. rhizogenes carrying pSV124 vector with ifn-α2b and nptII genes, as well as the roots of non-transformed plants that were cultivated in vitro. The presence and activity of rolB, rolC, PAL, and CHS genes were determined by PCR and real-time PCR, respectively. The content of flavonoids, AOA and RP were determined by standard tests with AlCl3, DPPH (2,2-diphenyl-1-picrylhydrazyl) and K3[Fe(CN)6] accordingly. Results. The content of flavonoids in most of the lines was higher than in the control, and correlated with AOA and RP. Roots No. 10 were characterized by the fastest growth, which coincided with higher activity of rolB and rolC genes. The activities of PAL and CHS in “hairy” roots were lower than those in non-transformed ones. Conclusions. Root lines carrying only rolB and rolC and lines with additional ifn-α2b and nptII genes had similar ranges of flavonoids concentration, AOA and RP levels that exceeded those in the control. The dependence of the root growth rate, and lack of the dependence of the flavonoids content with the activity of the rol genes were demonstrated. PAL activity inversely correlated with flavonoids content in all experimental lines, which may be the result of overproduction of compounds in transgenic roots.
{"title":"PECULIARITIES OF THE GROWTH OF Artemisia tilesii Ledeb. “HAIRY” ROOTS WITH DIFFERENT FOREIGN GENES","authors":"Т. А. Bohdanovych","doi":"10.15407/biotech15.03.013","DOIUrl":"https://doi.org/10.15407/biotech15.03.013","url":null,"abstract":"Aim. To compare Artemisia tilesii “hairy” root lines with different transferred genes in terms of the relationship between the total content of flavonoids, the levels of antioxidant activity (AOA) and reducing power (RP), as well as the activity of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), rolB and rolC genes. Methods. We compared the root lines Nos. 10 and 16, obtained by transformation with the wild Agrobacterium rhizogenes strain A4, lines Nos. 2 and 4, obtained using A. rhizogenes carrying pSV124 vector with ifn-α2b and nptII genes, as well as the roots of non-transformed plants that were cultivated in vitro. The presence and activity of rolB, rolC, PAL, and CHS genes were determined by PCR and real-time PCR, respectively. The content of flavonoids, AOA and RP were determined by standard tests with AlCl3, DPPH (2,2-diphenyl-1-picrylhydrazyl) and K3[Fe(CN)6] accordingly. Results. The content of flavonoids in most of the lines was higher than in the control, and correlated with AOA and RP. Roots No. 10 were characterized by the fastest growth, which coincided with higher activity of rolB and rolC genes. The activities of PAL and CHS in “hairy” roots were lower than those in non-transformed ones. Conclusions. Root lines carrying only rolB and rolC and lines with additional ifn-α2b and nptII genes had similar ranges of flavonoids concentration, AOA and RP levels that exceeded those in the control. The dependence of the root growth rate, and lack of the dependence of the flavonoids content with the activity of the rol genes were demonstrated. PAL activity inversely correlated with flavonoids content in all experimental lines, which may be the result of overproduction of compounds in transgenic roots.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43947896","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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