Pub Date : 2021-03-01DOI: 10.30491/JABR.2020.234981.1238
N. Ghorbani, M. Assmar, N. Amirmozafari, K. Issazadeh
Introduction: Globally, Salmonella enterica serotype Typhi is responsible for more than 10 million enteric fever cases, annually. Because of the emergence of multidrug-resistant strains of many bacteria, including Salmonella Typhi, vaccination may be a preferred strategy to combat infectious diseases. In the present study, the efficiency of flagellin protein as a recombinant vaccine candidate was evaluated in BALB/c mice. Materials and Methods: For this aim, flagellin protein was expressed in E. coli BL21 (DE3). Mice were grouped into two groups: test and control. The test group was immunized by the intraperitoneal administration of recombinant protein in combination with Freund's adjuvant. Following the completion of the immunization period, the mice were challenged by IP injection of 10 LD50 of live Salmonella Typhi and subsequent culture of their spleens and livers. Results: Flagellin protein expression was confirmed by SDS-PAGE and Western blotting. ELISA showed the proper stimulation of the humoral immunity of the immunized mice. The bacterial count decreased significantly in the spleens and livers of the immunized animals in comparison to the control ones. Conclusions: Findings of this study show the efficiency of flagellin recombinant protein in protecting mice against Salmonella Typhi.
{"title":"Recombinant Flagellin Protein Can Efficiently Protect Mice Against Salmonella Typhi","authors":"N. Ghorbani, M. Assmar, N. Amirmozafari, K. Issazadeh","doi":"10.30491/JABR.2020.234981.1238","DOIUrl":"https://doi.org/10.30491/JABR.2020.234981.1238","url":null,"abstract":"Introduction: Globally, Salmonella enterica serotype Typhi is responsible for more than 10 million enteric fever cases, annually. Because of the emergence of multidrug-resistant strains of many bacteria, including Salmonella Typhi, vaccination may be a preferred strategy to combat infectious diseases. In the present study, the efficiency of flagellin protein as a recombinant vaccine candidate was evaluated in BALB/c mice. Materials and Methods: For this aim, flagellin protein was expressed in E. coli BL21 (DE3). Mice were grouped into two groups: test and control. The test group was immunized by the intraperitoneal administration of recombinant protein in combination with Freund's adjuvant. Following the completion of the immunization period, the mice were challenged by IP injection of 10 LD50 of live Salmonella Typhi and subsequent culture of their spleens and livers. Results: Flagellin protein expression was confirmed by SDS-PAGE and Western blotting. ELISA showed the proper stimulation of the humoral immunity of the immunized mice. The bacterial count decreased significantly in the spleens and livers of the immunized animals in comparison to the control ones. Conclusions: Findings of this study show the efficiency of flagellin recombinant protein in protecting mice against Salmonella Typhi.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43716194","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 : 2021-03-01DOI: 10.30491/JABR.2021.127914
Nedda Hamidi, B. G. Eimani, M. Motamedi, Rouhollah Kazemi, A. Hajizade, J. Amani
Introduction: Enterotoxigenic E. coli (ETEC) is one of the major causes of watery diarrhea outbreaks in children under five years of age and passengers in developing countries. The pathogenicity of ETEC is due to the secretion of Colonization factors (Cfs) and two enterotoxins including heat-labile (Lt) and heat-stable (St). Although diarrhea is considered as one of the causes of mortality in developing countries, no approved vaccine is available for the disease caused by ETEC. Accordingly, the objective of the present study was to design a vaccine candidate containing Sta toxin which accounts for 30-75% of ETEC species. Materials and Methods: A chimeric construct consisting of two Sta toxoid connected together by two linkers was designed. After expression and purification by Ni-NTA column, western blotting was performed to confirm the protein. The 2Sta protein was administered to BALB/c mice via injection and the serum and fecal antibodies titer was evaluated by the ELISA test. Results: The recombinant 2Sta protein was expressed in an insoluble form (inclusion body) and the 20 kDa band was observed on the SDS-PAGE 12%. The results of the ELISA test suggested that IgG and IgA antibodies had enhanced compared to the control group. Conclusions: The Sta protein which was produced in most ETEC species can be induced in the immune system and raised the serum and fecal antibodies. Using this candidate subunit vaccine could actually protect bacterial infection.
{"title":"Design and Production of Engineered Sta Toxin as a Vaccine Candidate Against Enterotoxigenic Escherichia coli","authors":"Nedda Hamidi, B. G. Eimani, M. Motamedi, Rouhollah Kazemi, A. Hajizade, J. Amani","doi":"10.30491/JABR.2021.127914","DOIUrl":"https://doi.org/10.30491/JABR.2021.127914","url":null,"abstract":"Introduction: Enterotoxigenic E. coli (ETEC) is one of the major causes of watery diarrhea outbreaks in children under five years of age and passengers in developing countries. The pathogenicity of ETEC is due to the secretion of Colonization factors (Cfs) and two enterotoxins including heat-labile (Lt) and heat-stable (St). Although diarrhea is considered as one of the causes of mortality in developing countries, no approved vaccine is available for the disease caused by ETEC. Accordingly, the objective of the present study was to design a vaccine candidate containing Sta toxin which accounts for 30-75% of ETEC species. Materials and Methods: A chimeric construct consisting of two Sta toxoid connected together by two linkers was designed. After expression and purification by Ni-NTA column, western blotting was performed to confirm the protein. The 2Sta protein was administered to BALB/c mice via injection and the serum and fecal antibodies titer was evaluated by the ELISA test. Results: The recombinant 2Sta protein was expressed in an insoluble form (inclusion body) and the 20 kDa band was observed on the SDS-PAGE 12%. The results of the ELISA test suggested that IgG and IgA antibodies had enhanced compared to the control group. Conclusions: The Sta protein which was produced in most ETEC species can be induced in the immune system and raised the serum and fecal antibodies. Using this candidate subunit vaccine could actually protect bacterial infection.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45823799","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 : 2021-03-01DOI: 10.30491/JABR.2020.230689.1222
J. Rashidiani, Khadijeh Eskandari, R. Ranjbar, H. Kooshki, D. Afshar, F. S. Dehkordi
Introduction: For rapid and sensitive detection of Vibrio cholerae an accurate assay is needed. In the present study, a gold-coated magnetic nanoparticle (GMNP) was investigated for the detection of V. cholerae. Materials and Methods: GMNPs were synthesized and functionalized by 11-mercapto-undecanoic acid (MUA) as a linker for immobilization of IgG against V. cholerae OmpW antigen. In the next step, IgG was coupled with carboxylic group of MUA using EDC / NHS and the IgG/GMNPs nanocomposite created and finally, the bacterium was detected in a sandwich model ELISA. Results: The IgG/GMNPs nanocomposite could detect V.cholerae in a concentration range from 2.5×10-2 to 1.5 × 105 N/ml (number of V.cholerae per ml). The correlation coefficient was 0.99 and the detection limit was 16 N/ml. Conclusions: In this study, the procedure of antibody immobilization on magnetic nanoparticles was designed. So that, by using magnetic nanoparticles, the pre-concentration as a time-consuming step was removed and the sensitivity of V. cholerae determination was increased. Also, this method can be extended to detect other microorganisms.
{"title":"Application of Gold Core-Shell Magnetic Nanoparticles as an Immunosensor for the Detection of Vibrio cholerae","authors":"J. Rashidiani, Khadijeh Eskandari, R. Ranjbar, H. Kooshki, D. Afshar, F. S. Dehkordi","doi":"10.30491/JABR.2020.230689.1222","DOIUrl":"https://doi.org/10.30491/JABR.2020.230689.1222","url":null,"abstract":"Introduction: For rapid and sensitive detection of Vibrio cholerae an accurate assay is needed. In the present study, a gold-coated magnetic nanoparticle (GMNP) was investigated for the detection of V. cholerae. Materials and Methods: GMNPs were synthesized and functionalized by 11-mercapto-undecanoic acid (MUA) as a linker for immobilization of IgG against V. cholerae OmpW antigen. In the next step, IgG was coupled with carboxylic group of MUA using EDC / NHS and the IgG/GMNPs nanocomposite created and finally, the bacterium was detected in a sandwich model ELISA. Results: The IgG/GMNPs nanocomposite could detect V.cholerae in a concentration range from 2.5×10-2 to 1.5 × 105 N/ml (number of V.cholerae per ml). The correlation coefficient was 0.99 and the detection limit was 16 N/ml. Conclusions: In this study, the procedure of antibody immobilization on magnetic nanoparticles was designed. So that, by using magnetic nanoparticles, the pre-concentration as a time-consuming step was removed and the sensitivity of V. cholerae determination was increased. Also, this method can be extended to detect other microorganisms.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47182372","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 : 2021-03-01DOI: 10.30491/JABR.2020.229267.1217
Nour El Houda Bekkar, B. Meddah, Bahadır Keskin, P. Sonnet
Introduction: The aim of this study was to evaluate the chemical composition of Z. lotus (ZL) and R. chalepensis (RC) Essential Oils (EOs), the oral acute toxicity, influence on the gastrointestinal (GI) microbiota, and the in vivo anti-salmonellosis effect. Materials and Methods: The EOs were isolated using the steam distillation process, and bioactive components were identified by GC-MS analysis. Oral acute toxicity, influence on the GI flora composition, and the anti-salmonellosis effect were elucidated using in vivo methods on experimental animals. Results: The GC-MS allowed us to identify 33 and 58 components in Z. lotus and R. chalepensis, respectively. Di-isooctyl phthalate (89.857%) was found to be the major compound identified in ZL. The main compounds in RC were 2-undecanone (26.528 %) followed by 2-nonanone (13.404 %). The LD50 of EOs was found to be greater than 5000 mg/kg. Also, no negative influence to intestinal microbiota was detected. An important decrease in S. enterica ssp arizonae cells achieving a bactericidal effect was recorded in rats treated with the EOs of both plants at a dose of 400 mg/kg. In parallel, an important significant (p <0.05) increase in lymphocyte number was observed for all tested animals. A decrease in alkaline phosphatase (ALP), alanine aminotransferase(ALT), and aspartate aminotransferase (AST) levels were observed. Furthermore, a reduced blood sedimentation rate (ESR) was recorded in treated animals. Conclusions: The Z. lotus and R. chalepensis act effectively as anti-salmonellosis agents, which support the use of these plants to cure gastrointestinal infections.
{"title":"Oral Acute Toxicity, Influence on the Gastrointestinal Microbiota and In vivo Anti-Salmonellosis Effect of Zizyphus lotus (L.) and Ruta chalepensis (L.) Essential Oils","authors":"Nour El Houda Bekkar, B. Meddah, Bahadır Keskin, P. Sonnet","doi":"10.30491/JABR.2020.229267.1217","DOIUrl":"https://doi.org/10.30491/JABR.2020.229267.1217","url":null,"abstract":"Introduction: The aim of this study was to evaluate the chemical composition of Z. lotus (ZL) and R. chalepensis (RC) Essential Oils (EOs), the oral acute toxicity, influence on the gastrointestinal (GI) microbiota, and the in vivo anti-salmonellosis effect. Materials and Methods: The EOs were isolated using the steam distillation process, and bioactive components were identified by GC-MS analysis. Oral acute toxicity, influence on the GI flora composition, and the anti-salmonellosis effect were elucidated using in vivo methods on experimental animals. Results: The GC-MS allowed us to identify 33 and 58 components in Z. lotus and R. chalepensis, respectively. Di-isooctyl phthalate (89.857%) was found to be the major compound identified in ZL. The main compounds in RC were 2-undecanone (26.528 %) followed by 2-nonanone (13.404 %). The LD50 of EOs was found to be greater than 5000 mg/kg. Also, no negative influence to intestinal microbiota was detected. An important decrease in S. enterica ssp arizonae cells achieving a bactericidal effect was recorded in rats treated with the EOs of both plants at a dose of 400 mg/kg. In parallel, an important significant (p <0.05) increase in lymphocyte number was observed for all tested animals. A decrease in alkaline phosphatase (ALP), alanine aminotransferase(ALT), and aspartate aminotransferase (AST) levels were observed. Furthermore, a reduced blood sedimentation rate (ESR) was recorded in treated animals. Conclusions: The Z. lotus and R. chalepensis act effectively as anti-salmonellosis agents, which support the use of these plants to cure gastrointestinal infections.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45336828","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 : 2021-03-01DOI: 10.30491/JABR.2020.222547.1194
A. R. Ghara, Fereshteh Ezzati Ghadi, Seyed Hamzeh Hosseini, S. Piacente, A. Cerulli, A. Alizadeh, R. Mirmahmoudi
Introduction: The twig of Capparis decidua has been traditionally used as an antidiabetic agent but its medicinal use has not been scientifically proved yet. Present study evaluated the antidiabetic effect and antioxidant activities of aquatic extract of twigof Capparis decidua on liver and pancreas of diabetic rats induced by Streptozotocin (STZ). Materials and Methods: The effect of Capparis decidua was evaluated by biochemical, histological and FTIR studies. All biochemical and biophysical parameters were estimated after 15 days of daily oral administrations of the aqueous extract. Results: Results showed that oral use of Capparis decidua extract (250 mg/kg) caused significant reduction in the fasting blood glucose levels in diabetic rats when compared to control rats. Moreover, the altered level of lipid peroxidation, antioxidant, lipid profiles, liver enzymes, and also structural changes were reversed in STZ-induced diabetic rats which received Capparis decidua extract. Conclusions: In conclusion, aqueous extract of Capparis decidua has potent antidiabetic and antioxidant activity.
{"title":"Antioxidant and Antidiabetic Effect of Capparis decidua Edgew (Forssk.) Extract on Liver and Pancreas of Streptozotocin-Induced Diabetic Rats","authors":"A. R. Ghara, Fereshteh Ezzati Ghadi, Seyed Hamzeh Hosseini, S. Piacente, A. Cerulli, A. Alizadeh, R. Mirmahmoudi","doi":"10.30491/JABR.2020.222547.1194","DOIUrl":"https://doi.org/10.30491/JABR.2020.222547.1194","url":null,"abstract":"Introduction: The twig of Capparis decidua has been traditionally used as an antidiabetic agent but its medicinal use has not been scientifically proved yet. Present study evaluated the antidiabetic effect and antioxidant activities of aquatic extract of twigof Capparis decidua on liver and pancreas of diabetic rats induced by Streptozotocin (STZ). Materials and Methods: The effect of Capparis decidua was evaluated by biochemical, histological and FTIR studies. All biochemical and biophysical parameters were estimated after 15 days of daily oral administrations of the aqueous extract. Results: Results showed that oral use of Capparis decidua extract (250 mg/kg) caused significant reduction in the fasting blood glucose levels in diabetic rats when compared to control rats. Moreover, the altered level of lipid peroxidation, antioxidant, lipid profiles, liver enzymes, and also structural changes were reversed in STZ-induced diabetic rats which received Capparis decidua extract. Conclusions: In conclusion, aqueous extract of Capparis decidua has potent antidiabetic and antioxidant activity.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47732831","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 : 2021-03-01DOI: 10.30491/JABR.2020.237342.1246
A. Nakanishi, Yuri Sakihma, Nanami Ozawa
Introduction: Chlamydomonas reinhardtii produces lipid and carbohydrate as an industrially useful bioproduct with the supply of CO2 as a carbon source. The CO2 supplying system, especially aeration rate through the photobioreactor, should be controlled to enhance cell proliferation. Materials and Methods: After fixing CO2 concentration as 0.8%, the aeration rate was controlled to increase stepwisely by 10 mL·min-1, 20 mL·min-1, or 40 mL·min-1 beginning at 10 mL·min-1 to a maximum of 50 mL·min-1 after the pH ˃ 6.5. To show the effect of CO2-supply, the broth condition and the cell-component of lipid, carbohydrate, and protein were evaluated. Results: The CO2 supplying condition increasing by 10 mL·min-1 stepwisely when over pH 6.5 in 100 mL of broth led to rapid cell proliferation reached a plateau 2 days earlier than in other conditions. On the other hand, the cell components incubated under 10 mL·min-1 stepwise condition showed no difference among the other conditions. Conclusions: Cell proliferation was improved by optimized stepwise CO2 aeration rates versus cell concentration in broth, and cell components were not changed even with improved cell proliferation. According to the results, it could be possible to improve material productivity by increasing biomass productivity.
{"title":"Improvement of Growth of Chlamydomonas reinhardtii in CO2 – Stepwisely Aerating Condition","authors":"A. Nakanishi, Yuri Sakihma, Nanami Ozawa","doi":"10.30491/JABR.2020.237342.1246","DOIUrl":"https://doi.org/10.30491/JABR.2020.237342.1246","url":null,"abstract":"Introduction: Chlamydomonas reinhardtii produces lipid and carbohydrate as an industrially useful bioproduct with the supply of CO2 as a carbon source. The CO2 supplying system, especially aeration rate through the photobioreactor, should be controlled to enhance cell proliferation. Materials and Methods: After fixing CO2 concentration as 0.8%, the aeration rate was controlled to increase stepwisely by 10 mL·min-1, 20 mL·min-1, or 40 mL·min-1 beginning at 10 mL·min-1 to a maximum of 50 mL·min-1 after the pH ˃ 6.5. To show the effect of CO2-supply, the broth condition and the cell-component of lipid, carbohydrate, and protein were evaluated. Results: The CO2 supplying condition increasing by 10 mL·min-1 stepwisely when over pH 6.5 in 100 mL of broth led to rapid cell proliferation reached a plateau 2 days earlier than in other conditions. On the other hand, the cell components incubated under 10 mL·min-1 stepwise condition showed no difference among the other conditions. Conclusions: Cell proliferation was improved by optimized stepwise CO2 aeration rates versus cell concentration in broth, and cell components were not changed even with improved cell proliferation. According to the results, it could be possible to improve material productivity by increasing biomass productivity.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49547178","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 : 2021-03-01DOI: 10.30491/JABR.2021.128849
M. Mobarakian, M. N. Dastjerdi, J. Shakarami, S. M. Abtahi
Introduction: To the best of our knowledge, there is little information in regards to the cytotoxicity of 1,8- cineole. Insect cells exhibit wide resistance to the lethal effects of chemical compounds. Activation of p53 by acetylation can cause cell toxicity. This study has been carried out in order to investigate whether the use of 1,8-cineole in the Spodoptera frugiperda (SF-9) cell line can induce cytotoxicity by increasing p53 acetylation expression or not. Materials and Methods: SF-9 cell line was cultured in TC100 insect culture medium and treated with 1,8-cineole at concentration of 850.45 μmol/L, based on the half-maximal inhibitory concentration (IC50) index at different times (24, 48, and 72h). The IC50 value was estimated for 1,8-cineole in SF-9. The percentage of alive cells was measured by MTT assay. The ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in SF-9 cells. Results: The findings of the present study indicated that treatment with 850.45 μmol per liter of 1,8-cineole shows a time-dependent increase in the number of dead cells of SF-9 cell line. The effect of severe toxicity on SF-9 was observed after 72 h of incubation with 1,8-cineole, with approximately 4% of SF-9 cells alive. We observed a significant increase in the level of p53 acetylation up to 48 h in SF-9, indicating that 1,8-cineole resulted in up‑regulation of acetylated P53 and consequently p53 activation in SF-9 cells. Results showed that there is relationship between acetylated p53 protein levels and 1,8-cineole toxicity in SF-9 cell line. Conclusions: 1,8-cineole, may function through common pathways and mediate their cytotoxic effects through targeting p53 and its acetylation in SF-9 cells.
{"title":"Mechanisms of Cytotoxicity-Inducing Effect of 1,8-cineole, a Plant Terpenoid, on Lepidopteran (Spodoptera frugiperda) Cell Line","authors":"M. Mobarakian, M. N. Dastjerdi, J. Shakarami, S. M. Abtahi","doi":"10.30491/JABR.2021.128849","DOIUrl":"https://doi.org/10.30491/JABR.2021.128849","url":null,"abstract":"Introduction: To the best of our knowledge, there is little information in regards to the cytotoxicity of 1,8- cineole. Insect cells exhibit wide resistance to the lethal effects of chemical compounds. Activation of p53 by acetylation can cause cell toxicity. This study has been carried out in order to investigate whether the use of 1,8-cineole in the Spodoptera frugiperda (SF-9) cell line can induce cytotoxicity by increasing p53 acetylation expression or not. Materials and Methods: SF-9 cell line was cultured in TC100 insect culture medium and treated with 1,8-cineole at concentration of 850.45 μmol/L, based on the half-maximal inhibitory concentration (IC50) index at different times (24, 48, and 72h). The IC50 value was estimated for 1,8-cineole in SF-9. The percentage of alive cells was measured by MTT assay. The ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in SF-9 cells. Results: The findings of the present study indicated that treatment with 850.45 μmol per liter of 1,8-cineole shows a time-dependent increase in the number of dead cells of SF-9 cell line. The effect of severe toxicity on SF-9 was observed after 72 h of incubation with 1,8-cineole, with approximately 4% of SF-9 cells alive. We observed a significant increase in the level of p53 acetylation up to 48 h in SF-9, indicating that 1,8-cineole resulted in up‑regulation of acetylated P53 and consequently p53 activation in SF-9 cells. Results showed that there is relationship between acetylated p53 protein levels and 1,8-cineole toxicity in SF-9 cell line. Conclusions: 1,8-cineole, may function through common pathways and mediate their cytotoxic effects through targeting p53 and its acetylation in SF-9 cells.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41744718","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 : 2021-03-01DOI: 10.30491/JABR.2021.128739
F. Zafari, M. Sadeghi
Introduction: Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinase. Several studies have reported the association between the single nucleotide polymorphisms(C/T) -1562 in the MMP-9 promoter and the risk of cancer. In this study, we decided to carry out a comprehensive meta-analysis to obtain a reasonable result about the association between this polymorphism and the risk of lung cancer. Materials and Methods: A complete literature review was conducted within the databases of ISI Web of Knowledge, google scholar, and PubMed for studies on lung cancer published from 2002 to 2018. A meta-analysis was conducted which included more than 3000 case and control subjects. The pooled Odds Ratio (OR) and 95% Confidence Intervals ( CI) were used for dominant, recessive, and co-dominant MMP-9 genotypes to assess the strength of the association. Results: Analysis indicated no significant association between MMP-9-1562C/T polymorphism and the risk of lung cancer, dominant model; [CT+TT/CC]: OR = 0.972, 95% CI = 0.811–1.164, recessive model [CC+CT/TT] OR= 1.027, 95% CI = 0.651–1.618 and co-dominant model [TT/CC] OR = 0.983, 95% CI = 0.550–1.755. Conclusions: No association was observed between the MMP9 -1562 C/T polymorphism and the incidence of lung cancer. The meta-analysis demonstrated that this polymorphism can't serve as a diagnostic marker for lung cancer.
引言：基质金属蛋白酶（MMPs）是一个锌依赖性蛋白酶家族。一些研究报告了MMP-9启动子中的单核苷酸多态性（C/T）-1562与癌症风险之间的关系。在这项研究中，我们决定进行一项全面的荟萃分析，以获得关于这种多态性与癌症风险之间关系的合理结果。材料和方法：在ISI Web of Knowledge、google scholar和PubMed的数据库中对2002年至2018年发表的癌症研究进行了完整的文献综述。进行了一项荟萃分析，包括3000多名病例和对照受试者。对显性、隐性和共显性MMP-9基因型使用合并比值比（OR）和95%置信区间（CI）来评估相关性的强度。结果：分析显示MMP-9-1562C/T多态性与癌症患病风险无显著相关性，为优势模型；[CT+TT/CC]：OR=0.972，95%CI=0.811–1.164，隐性模型[CT+CT/TT]OR=1.027，95%CI=0.651–1.618，共显性模型[TT/CC]OR=0.983，95%CI 0.550–1.755。结论：MMP9-1562 C/T多态性与癌症发病率无相关性。荟萃分析表明，这种多态性不能作为癌症的诊断标志。
{"title":"The Association Between Matrix Metalloproteinases-9-1562 C/T Polymorphism and Lung Cancer Susceptibility: A Systematic Review and Meta-Analysis","authors":"F. Zafari, M. Sadeghi","doi":"10.30491/JABR.2021.128739","DOIUrl":"https://doi.org/10.30491/JABR.2021.128739","url":null,"abstract":"Introduction: Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinase. Several studies have reported the association between the single nucleotide polymorphisms(C/T) -1562 in the MMP-9 promoter and the risk of cancer. In this study, we decided to carry out a comprehensive meta-analysis to obtain a reasonable result about the association between this polymorphism and the risk of lung cancer. Materials and Methods: A complete literature review was conducted within the databases of ISI Web of Knowledge, google scholar, and PubMed for studies on lung cancer published from 2002 to 2018. A meta-analysis was conducted which included more than 3000 case and control subjects. The pooled Odds Ratio (OR) and 95% Confidence Intervals ( CI) were used for dominant, recessive, and co-dominant MMP-9 genotypes to assess the strength of the association. Results: Analysis indicated no significant association between MMP-9-1562C/T polymorphism and the risk of lung cancer, dominant model; [CT+TT/CC]: OR = 0.972, 95% CI = 0.811–1.164, recessive model [CC+CT/TT] OR= 1.027, 95% CI = 0.651–1.618 and co-dominant model [TT/CC] OR = 0.983, 95% CI = 0.550–1.755. Conclusions: No association was observed between the MMP9 -1562 C/T polymorphism and the incidence of lung cancer. The meta-analysis demonstrated that this polymorphism can't serve as a diagnostic marker for lung cancer.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49145457","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 : 2021-03-01DOI: 10.30491/JABR.2020.226481.1210
A. Estaji, E. Chamani, Z. khazaei
Introduction: Lily is often described as one of the most widespread, commercial crops in the floriculture industry. Commercially grown cultivars are mostly propagated by bulb scales which it cost effective and uniformity in tissue culture conditions. Plant tissue culture techniques can effectively provide far-reaching implications in micro-propagation. Materials and Methods: This study was conducted to investigate the effects of various concentrations of cytokines and auxins on callogenesis and Biomass under in vitro conditions. In this experiment, the evaluations were aimed at measuring different characteristics in two lily cultivars, namely, Lilium ledebourii and Lilium regal. Results: The results showed significant values in all of the measured characteristics. The highest percentage of callogenesis was caused by 2 µm PIC plus 1 µm KIN in L. ledebourii (88.66%) and L. regal (88.66%). Also, the callus weight in both cultivars was obtained by applying the same combination of treatments. In the second experiment, the highest fresh biomass of the cell suspension occurred by applying 2 µm PIC plus 4 µm KIN. The maximum amount of fresh biomass in L. ledebourii (88.93 g/L) and L. regal (41 g/L) occurred on the 24th and 20th day of the culture, respectively. Conclusions: An efficient and fragile callus induction was developed by NAA, which nonetheless made the condition more suitable for cell suspension culture. These lily cultivars need high amounts of PIC as auxin to grow well in cell suspensions. By increasing the PIC level, the biomass accumulates more. These results can moderately optimize large-scale production of both fragile calli and cell suspension biomass.
{"title":"Influence of Plant Growth Regulators on Callogenesis and the Biomass of Cell Suspensions in Lily (Lilium ledebourii and Lilium regal)","authors":"A. Estaji, E. Chamani, Z. khazaei","doi":"10.30491/JABR.2020.226481.1210","DOIUrl":"https://doi.org/10.30491/JABR.2020.226481.1210","url":null,"abstract":"Introduction: Lily is often described as one of the most widespread, commercial crops in the floriculture industry. Commercially grown cultivars are mostly propagated by bulb scales which it cost effective and uniformity in tissue culture conditions. Plant tissue culture techniques can effectively provide far-reaching implications in micro-propagation. Materials and Methods: This study was conducted to investigate the effects of various concentrations of cytokines and auxins on callogenesis and Biomass under in vitro conditions. In this experiment, the evaluations were aimed at measuring different characteristics in two lily cultivars, namely, Lilium ledebourii and Lilium regal. Results: The results showed significant values in all of the measured characteristics. The highest percentage of callogenesis was caused by 2 µm PIC plus 1 µm KIN in L. ledebourii (88.66%) and L. regal (88.66%). Also, the callus weight in both cultivars was obtained by applying the same combination of treatments. In the second experiment, the highest fresh biomass of the cell suspension occurred by applying 2 µm PIC plus 4 µm KIN. The maximum amount of fresh biomass in L. ledebourii (88.93 g/L) and L. regal (41 g/L) occurred on the 24th and 20th day of the culture, respectively. Conclusions: An efficient and fragile callus induction was developed by NAA, which nonetheless made the condition more suitable for cell suspension culture. These lily cultivars need high amounts of PIC as auxin to grow well in cell suspensions. By increasing the PIC level, the biomass accumulates more. These results can moderately optimize large-scale production of both fragile calli and cell suspension biomass.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42603556","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 : 2021-02-03DOI: 10.30491/JABR.2020.223551.1199
Mohsen Norouzi, M. Rafienia, E. Poorazizi, M. Setayeshmehr
Introduction: A synthetic biomaterial, such as poly(lactic-co-glycolic acid) (PLGA) with superior mechanical properties, along with a natural polymer such as fibrin, which facilitates cell attachment and enhances biocompatibility, can be used in the production of novel composite tissue engineering scaffolds. In this study, we fabricated a PLGA/Fibrin scaffold which was evaluated to the enhancement of adipose-derived stem cells' growth and proliferation. Materials and Methods: 10% polymer solutions with different ratios of PLGA:Fibrin, including 10:0, 9:1, 8:2, and 7:3, were prepared and used in the production of aligned and unaligned electrospun nanofiber scaffolds. Human adipose-derived stem cells (h-ADSCs) were cultured on the scaffolds and they were characterized using FTIR, EDX, SEM, mechanical, hydrophilic, degradation, water absorption, and biocompatibility tests. Results: The obtained scaffolds consisted of homogeneous fibers, without any beads and water droplets. The percentage of porosities and internal correlation of the cavities are not significantly different between aligned and unaligned electrospun scaffolds (P>0.05) and by adding fibrin, these properties are improved, while tensile strength and elasticity are decreased. All the scaffolds are hydrophobic and the highest and lowest swelling rates belong to PLGA/30%Fibrin scaffolds and pure PLGA scaffolds (more than 90% and less than 45%, respectively). There is a significant difference in degradation rates between fibrin-contained scaffolds and pure PLGA scaffolds. Moreover, compared with aligned electrospun scaffolds, a higher degradation rate of unaligned ones was observed. Conclusions: Considering the results of SEM and bio-compatibility experiments, the aligned electrospun PLGA/10%Fibrin scaffold with numerous spindle shape h-ADSCs and unaligned electrospun PLGA/20%Fibrin scaffold with lots of spindle shape cells together with sort of round shape cells are introduced as the optimal options.
{"title":"Adipose-Derived Stem Cells Growth and Proliferation Enhancement Using Poly(Lactic-co-Glycolic Acid) (PLGA)/Fibrin Nanofiber Mats","authors":"Mohsen Norouzi, M. Rafienia, E. Poorazizi, M. Setayeshmehr","doi":"10.30491/JABR.2020.223551.1199","DOIUrl":"https://doi.org/10.30491/JABR.2020.223551.1199","url":null,"abstract":"Introduction: A synthetic biomaterial, such as poly(lactic-co-glycolic acid) (PLGA) with superior mechanical properties, along with a natural polymer such as fibrin, which facilitates cell attachment and enhances biocompatibility, can be used in the production of novel composite tissue engineering scaffolds. In this study, we fabricated a PLGA/Fibrin scaffold which was evaluated to the enhancement of adipose-derived stem cells' growth and proliferation. Materials and Methods: 10% polymer solutions with different ratios of PLGA:Fibrin, including 10:0, 9:1, 8:2, and 7:3, were prepared and used in the production of aligned and unaligned electrospun nanofiber scaffolds. Human adipose-derived stem cells (h-ADSCs) were cultured on the scaffolds and they were characterized using FTIR, EDX, SEM, mechanical, hydrophilic, degradation, water absorption, and biocompatibility tests. Results: The obtained scaffolds consisted of homogeneous fibers, without any beads and water droplets. The percentage of porosities and internal correlation of the cavities are not significantly different between aligned and unaligned electrospun scaffolds (P>0.05) and by adding fibrin, these properties are improved, while tensile strength and elasticity are decreased. All the scaffolds are hydrophobic and the highest and lowest swelling rates belong to PLGA/30%Fibrin scaffolds and pure PLGA scaffolds (more than 90% and less than 45%, respectively). There is a significant difference in degradation rates between fibrin-contained scaffolds and pure PLGA scaffolds. Moreover, compared with aligned electrospun scaffolds, a higher degradation rate of unaligned ones was observed. Conclusions: Considering the results of SEM and bio-compatibility experiments, the aligned electrospun PLGA/10%Fibrin scaffold with numerous spindle shape h-ADSCs and unaligned electrospun PLGA/20%Fibrin scaffold with lots of spindle shape cells together with sort of round shape cells are introduced as the optimal options.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-02-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43853967","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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