Background: Tuberculosis (TB) with more than 10 million new cases per year and one of the top 10 causes of death worldwide, is still one of the most important global health problems. Also, multi drug-resistant tuberculosis (MDR) is a serious danger to public health. Understanding of the epidemiological pattern of mycobacterium tuberculosis (MTB), Estimates of recent transmission and recurrence of infection, it is possible with molecular typing methods. The present study was performed aiming to track and determine the type of Mycobacterium tuberculosis infection, as well as its relationship with demographic factors, using PGRS-RFLP. Materials and Methods: In this study, 90 samples collected from TB patients from the North West provinces of Iran, Molecular typing by Characterization highly abundant polymorphic GC-rich-repetitive sequence. Investigated Demographic factors associated with the transmission of the disease. Results: All isolates were grouped into 44 clusters 28types (33.3%) of the subspecies were in unique stains and 66.7% (from 56 patients) had clustered isolates. The largest cluster contained 8 isolates (9.52%) was the West provinces of Iran. Conclusion: Genetic variation of Mycobacterium tuberculosis is high in this region. The rate of recent transmission based on clustering was unexpected (The global average is 30%-40%). The recent transmission was more dynamic in the west than northwest Iran. Clustering based on PGRS-RFLP can demonstrate the high correlation between molecular and classic information. In addition, the significant relationship between vaccination record and clustering highlights the necessity to conduct more extensive studies.
{"title":"Molecular Typing of Mycobacterium Tuberculosis Isolated from Iranian Patients Using Highly Abundant Polymorphic GC-Rich-Repetitive Sequence","authors":"B. G. Eimani, K. Ansarin, L. Sahebi, M. Seyyedi","doi":"10.52547/ismj.23.2.87","DOIUrl":"https://doi.org/10.52547/ismj.23.2.87","url":null,"abstract":"Background: Tuberculosis (TB) with more than 10 million new cases per year and one of the top 10 causes of death worldwide, is still one of the most important global health problems. Also, multi drug-resistant tuberculosis (MDR) is a serious danger to public health. Understanding of the epidemiological pattern of mycobacterium tuberculosis (MTB), Estimates of recent transmission and recurrence of infection, it is possible with molecular typing methods. The present study was performed aiming to track and determine the type of Mycobacterium tuberculosis infection, as well as its relationship with demographic factors, using PGRS-RFLP. Materials and Methods: In this study, 90 samples collected from TB patients from the North West provinces of Iran, Molecular typing by Characterization highly abundant polymorphic GC-rich-repetitive sequence. Investigated Demographic factors associated with the transmission of the disease. Results: All isolates were grouped into 44 clusters 28types (33.3%) of the subspecies were in unique stains and 66.7% (from 56 patients) had clustered isolates. The largest cluster contained 8 isolates (9.52%) was the West provinces of Iran. Conclusion: Genetic variation of Mycobacterium tuberculosis is high in this region. The rate of recent transmission based on clustering was unexpected (The global average is 30%-40%). The recent transmission was more dynamic in the west than northwest Iran. Clustering based on PGRS-RFLP can demonstrate the high correlation between molecular and classic information. In addition, the significant relationship between vaccination record and clustering highlights the necessity to conduct more extensive studies.","PeriodicalId":14583,"journal":{"name":"Iranian South Medical Journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43802131","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}
M. Fouladvand, Solyman Khorami, Behroz Naeimi, Sohila Fotouhi, Khosro Mohammadi
Background: Leishmania major is transmitted by sandflies of the genus Phobotomus and causes cutaneous lesions in humans. Curcumin is made from turmeric and various derivatives is derived from it. In the present study the effect of curcumin and it’s derivatives on Leishmania major in vitro was investigated. Materials and Methods: Curcumin 70% and 90% purity and it’ derivatives such as s , BDMC base methoxy curcumin, diacetyl curcumin DAC, vanadyl curcumin VO (CUR) 2, vanadyl diacetyl curcumin VO (DAC) 2, indium curcumin In(CUR)3 and Ga (CUR) 3 preparation. Curcumin derivatives were synthesized and different concentrations of 500 to 800 μg/ ml were prepared in glycerin. 106 Leishmania major promastigotes added to each well of 96 well plate and 100 µL of each curcumin derivative added and stored at 25 ° C for 24 hours. Assay of the lethality of the compounds against Leishmania major and evaluation of their toxic effects on (vero) cells were evaluated using MTT assay. Results: Lethal effect of curcumin and it’s derivatives against Leishmania, increased with increasing concentration. Anti-leishmanial index (IC 50 ) of 70% and 90% curcumin compounds, (BDMC) bis methoxy curcumin, Diacetyl curcumin (DAC), vanadyl curcumin VO (CUR)2, vanadyl diacetyl curcumin VO (DAC)2, indium curcumin In (CUR)3, Gallium Curcumin Ga (CUR)3 compounds were 110, 93, 101, 103, 98, 103, 51 and 58 µg / ml and the lethal effect of these compounds against (vero) cells were 33.1%, 19%, 21%, 20.3%, 17%, 21%, 25.3%, 16%, respectively. Conclusion: Gallium curcumin and indium curcumin, compared to curcumin and other derivatives, exhibited the highest anti-leishmanial effect and were the safest derivatives of curcumin for mammalian cells.
{"title":"Evaluation of Lethal Effect of Curcumin and its Derivatives Against Leishmania Major In Vitro","authors":"M. Fouladvand, Solyman Khorami, Behroz Naeimi, Sohila Fotouhi, Khosro Mohammadi","doi":"10.52547/ismj.23.2.153","DOIUrl":"https://doi.org/10.52547/ismj.23.2.153","url":null,"abstract":"Background: Leishmania major is transmitted by sandflies of the genus Phobotomus and causes cutaneous lesions in humans. Curcumin is made from turmeric and various derivatives is derived from it. In the present study the effect of curcumin and it’s derivatives on Leishmania major in vitro was investigated. Materials and Methods: Curcumin 70% and 90% purity and it’ derivatives such as s , BDMC base methoxy curcumin, diacetyl curcumin DAC, vanadyl curcumin VO (CUR) 2, vanadyl diacetyl curcumin VO (DAC) 2, indium curcumin In(CUR)3 and Ga (CUR) 3 preparation. Curcumin derivatives were synthesized and different concentrations of 500 to 800 μg/ ml were prepared in glycerin. 106 Leishmania major promastigotes added to each well of 96 well plate and 100 µL of each curcumin derivative added and stored at 25 ° C for 24 hours. Assay of the lethality of the compounds against Leishmania major and evaluation of their toxic effects on (vero) cells were evaluated using MTT assay. Results: Lethal effect of curcumin and it’s derivatives against Leishmania, increased with increasing concentration. Anti-leishmanial index (IC 50 ) of 70% and 90% curcumin compounds, (BDMC) bis methoxy curcumin, Diacetyl curcumin (DAC), vanadyl curcumin VO (CUR)2, vanadyl diacetyl curcumin VO (DAC)2, indium curcumin In (CUR)3, Gallium Curcumin Ga (CUR)3 compounds were 110, 93, 101, 103, 98, 103, 51 and 58 µg / ml and the lethal effect of these compounds against (vero) cells were 33.1%, 19%, 21%, 20.3%, 17%, 21%, 25.3%, 16%, respectively. Conclusion: Gallium curcumin and indium curcumin, compared to curcumin and other derivatives, exhibited the highest anti-leishmanial effect and were the safest derivatives of curcumin for mammalian cells.","PeriodicalId":14583,"journal":{"name":"Iranian South Medical Journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41627390","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}
Background: Molecular methods are nowadays used to diagnose diseases, produce vaccines, drugs and recombinant proteins. Therefore, one of the important steps in these procedures is selecting the appropriate vector for cloning and expression of the target genes. Because the expression of synthetic polygenes or so-called chimeric antigens with high molecular weight requires special conditions, the aim of this study was to evaluate the importance of vector type for expression of such antigens. Materials and Methods: The 1250bp sequence consisting of epitopes from eight important antigens of Leishmania infantum parasite was designed and synthesized by Biomatic Company (Cambridge, Canada). The sequence was cloned separately in two expression vectors pET25b and pET32a and then transformed into E.coli BL21 (DE3) and expressed under similar appropriate conditions. Bacterial lysates were analyzed by SDS PAGE and evaluated by Western blotting. Results: of SDS- PAGE and Western blot analysis showed that intact recombinant protein production by pET25b vector was not successful. However, the recombinant protein resulted from the expression of the aforementioned poly-epitope in pET32a vector was successfully produced and confirmed. Conclusion: According to the successful expression confirmation of the poly-epitopic sequence within the pET32a vector and furthermore the failure to obtain protein in the pET25b vector showed that in the case of some specific mosaic sequences expression, protein isolation will be difficult because of low solubility. So, the expression vector choose should be made more carefully.
{"title":"Comparative Evaluation of Expression Vectors (pET32a and pET25b) in Expression of Polyepitopic Sequence Synthesized from \u0000Leishmania infantum Antigens","authors":"Marzeah Taherzadeh, M. Fouladvand, B. Kazemi","doi":"10.52547/ismj.23.2.143","DOIUrl":"https://doi.org/10.52547/ismj.23.2.143","url":null,"abstract":"Background: Molecular methods are nowadays used to diagnose diseases, produce vaccines, drugs and recombinant proteins. Therefore, one of the important steps in these procedures is selecting the appropriate vector for cloning and expression of the target genes. Because the expression of synthetic polygenes or so-called chimeric antigens with high molecular weight requires special conditions, the aim of this study was to evaluate the importance of vector type for expression of such antigens. Materials and Methods: The 1250bp sequence consisting of epitopes from eight important antigens of Leishmania infantum parasite was designed and synthesized by Biomatic Company (Cambridge, Canada). The sequence was cloned separately in two expression vectors pET25b and pET32a and then transformed into E.coli BL21 (DE3) and expressed under similar appropriate conditions. Bacterial lysates were analyzed by SDS PAGE and evaluated by Western blotting. Results: of SDS- PAGE and Western blot analysis showed that intact recombinant protein production by pET25b vector was not successful. However, the recombinant protein resulted from the expression of the aforementioned poly-epitope in pET32a vector was successfully produced and confirmed. Conclusion: According to the successful expression confirmation of the poly-epitopic sequence within the pET32a vector and furthermore the failure to obtain protein in the pET25b vector showed that in the case of some specific mosaic sequences expression, protein isolation will be difficult because of low solubility. So, the expression vector choose should be made more carefully.","PeriodicalId":14583,"journal":{"name":"Iranian South Medical Journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46180139","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}