Tarbiat Modares UniversityPathobiology Research2538-300012320090901Effect of protein and DNA components of Toxoplasma gondii on IL-12 production from dendritic cells and T cell proliferationمقایسه تأثیر اجزای پروتئینی و اسید نوکلئیکی عصاره توکسوپلاسما گوندی بر تولید IL-12 از سلولهای دندریتیک و تکثیر سلولهای T1819006ENJamshidHadjatiDepartment of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, IranافشینآماریAliRezaRazaviDepartment of Immunology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, IranArezooJamaliDepartment of Immunology, Faculty of Paramedicene , Tehran University of Medical Sciences, Tehran, IranAbbasAliAmini SardrodDepartment of Immunology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, IranMasomehMotamediDepartment of Immunology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, IranSaeedehShojaeeDepartment of Parasithology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, IranBitaAnsaripourDepartment of Immunology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, IranJournal ArticleObjective: The aim of the present study was to investigate the effect of protein and DNA components of Toxoplasma gondii on maturation of dendritic cells and their efficiency in IL-12 production and proliferation of T cells.
Materials and Methods: for DC generation, Bone marrow cells were cultured in the presence of GM-CSF and IL-4 for 5 days. Tumor lysate and protein or DNA components of Toxoplasma gondii were added to the culture media and incubated for another 2 days. LPS was added as control for DC maturation. Proliferation of T cells were determined by MLR and IL-12 production was measured by ELISA kit. Maturation of dendritic cell were determined by flowcytometry.
Results: DCs treatment with protein components of Toxoplasma gondii caused a significant increase in IL-12 production and proliferation of T cells (P<0.001).
Conclusions: Different compositions of microbial body like protein and DNA components of Toxoplasma gondii can cause augmentation of antigen presentation capacity of DC and their IL-12 production capability. Among these components the protein was more effective as compared to DNA.https://mjms.modares.ac.ir/article_19006_a56b6ea9b986428c2ab2fb458f5da882.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Evaluation of PCR and Xenodiagnosis assays for detection of Borrelia persica in soft ticks Ornithodoros tholozaniارزیابی روشهای مولکولی PCR و Xenodiagnosis برای تعیین آلودگی کنههای نرم اورنیتودوروس تولوزانی به بورلیا پرسیکا91619007ENMohammad AliOshaghiDepartment of Medical Entomology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranJavadRafinejadDepartment of Medical Entomology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranNayerehChoubdarDepartment of Medical Entomology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranAlirezaBarmakiDepartment of Medical Entomology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranNorayerPiazakDepartment of Parasitology, Iran Institute of Pasteur, Tehran, IranFatemehMohtaramiDepartment of Medical Entomology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranTaghiSatvatDepartment of Parasitology, Faculty of Public Health and Institute of Health Research (SPH & IHR), Tehran University of Medical Sciences, Tehran, IranOmidBanafshiDepartment of Health Management, Kermanshah University of Medical Sciences, Kermanshah, IranBehrozTaghilooDepartment of Health Management, Zanjan University of Medical Sciences, Zanjan, IranNasehMaleki RavasanParasitology & Medical Entomology, School of Medical Sciences, Tarbiat Modares University, Tehran, IranJournal ArticleObjective: Relapsing fever caused by Borrelia persica is an acute tick-borne disease which is transmitted by soft ticks of Ornithodoros tholozani to human. The disease is reported from Middle East and many regions of Iran. Detection of infection is problematic since the suspected infected ticks should be fed on animal hosts such as guinea pigs and subsequently after 7-14 days, the animal blood should be microscopically investigated for Borellia spirochetes on a Giemsa stainined thick smear. This classic method named xenodiagnosis is hard, time consuming, and less reliable. In this study, the application of PCR technique has been examined for detection of Borellia persica in soft ticks of O. tholozani.
Materials and Methods: Tick specimens were collected from northwestern Iran and were fed on Borellia persica infected guinea pigs. DNA of the animal blood were extracted and used as target for PCR amplification of 16rDNA gene. Subsequently the products were subjected to sequencing. The effect of tick sex and post digestion as well as the minimum number of spirochetes on the efficiency of PCR were also tested.
Results: The xenodiagnosis assay was able to detect infection in only 13.3% of the tick-bitten animal bloods whereas all of these blood specimens were PCR positive against the 16rDNA gene. There wasno difference in results of PCR for male and female of the ticks. Post digestion of infected blood meal in ticks did not affect the efficacy of PCR and the recently-fed samples showed similar results to those of completely gravid ones. A test on the threshold sensitivity of PCR assay indicated that only one spirochete is enough for the primers to anneal and to amplify the target gene.
Conclusion: This study describes the first molecular assay for diagnosis of B. persica infected ticks in Iran and due to its high speed, accuracy, and applicability is a substitution method for diagnostic purposes in TBRF foci.https://mjms.modares.ac.ir/article_19007_4ef124d6f895ab6b274819566be43b12.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Use of Real-Time PCR method on the amniocyte samples for prenatal diagnosis of Down syndromeاستفاده از روش Real-Time PCR روی نمونههای آمنیوسیت برای تشخیص پیش از تولد نشانگان داون173119008ENRezaMahdianDepartment of Medical Biotechnology, Pasteur Institute of Iran, Tehran, IranFahimehShahrokhiDepartment of Human Genetics, Faculty of Medicine, Tabriz University of Medical Science, Tabriz, IranMojtabaMohaddes ArdebiliDepartment of Medical Genetics, Faculty of Medicine, Tabriz University of Medical Science, Tabriz, IranZahraFardi AzarDepartment of Gynecology, Faculty of Medicine, Tabriz University of Medical Science, Tabriz, IranAhmadKamyabDepartment of Human Genetics, Pasteur Institute of Iran, Tehran, IranElmiraShamsiyanDepartment of Human Genetics, Pasteur Institute of Iran, Tehran, IranMinaHayat NosaeidDepartment of Molecular Medicine, Pasteur Institute of Iran, Tehran, IranZahraKaeeni MoghaddamDepartment of Molecular Medicine, Pasteur Institute of Iran, Tehran, IranJournal ArticleObjective: In this study, the possibility of prenatal diagnosis of Down syndrome with Real-Time PCR method was evaluated. In this context, optimization of a suitable method for purification of high quality DNA from amniotic fluid samples was also considered.
Materials and Methods: Pregnant women who had the high risk of having babies with Down syndrome were selected according to the biochemical and sonographic data and referred to the amniocentesis center. The DNA of total 59 amniotic fluid samples were extracted with different methods including boiling method, salting out method, Procedures of DNA extraction from Blood and Cell Culture by DNPTM Kit (CinnaGen), Procedure of DNA extraction from cells by DNA Isolation Kit for cells and tissues (Roche), Procedure of DNA extraction from Tissue by MagNa Pure DNA Isolation kit (Roche), and QIAamp DNA Micro Kit (Qiagen). Then, the quality and quantity of the extracted DNA were evaluated by the NanoDrop® ND- 1000 spectrophotometer device. Real-Time PCR reaction using fluorescent dye SYBR Green I (Applied Biosystems, UK) was performed to specifically amplify DSCAM and DYRK1A2 genes and the reference gene (PMP22). Data analysis was performed using comparative cycle threshold method for the determination of the gene dosage and determining the number of copies of chromosome 21.
Results: This study showed that DNA extracted from amniotic fluid samples using QIAamp DNA Micro Kit (Qiagen) has the desirable quantity and quality for Real-Time PCR. Specific proliferation of targets and reference genes was achieved and difference between normal and affected groups based on differences between their gene dosages was determined.
Conclusion: Prenatal diagnosis of Down syndrome is feasible by the Real-Time PCR method using DNA samples from amniotic fluid cells extracted by QIAamp DNA Micro Kit (Qiagen). The results are comparable to the corresponding results from conventional cytogenetic methods.https://mjms.modares.ac.ir/article_19008_6c3a4ee6624ec8f6a1700123174d614e.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Designing and development of a silencer vector based on second generation of RNA interference (RNAi) technique to study downregulation of steroid receptor RNA activator (SRA)طراحی و ساخت ناقل خاموشگر RNA تداخلگر (RNAi) نسل دوم برای بررسی کاهش بیان ژن کمک فعالکننده RNA گیرنده استروئیدی (SRA)333919009ENMehdiForouzandehDepartment of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modares University, Tehran, IranSaeedehAskarianDepartment of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modares University, Tehran, IranلیدالنگرودیفاطمهرهبریزادهJournal ArticleObjective: RNA interference (RNAi) is the most potent technique for gene silencing in eukaryotic cellular system at transcriptomic level. Genetic disorders and cancers are important targets for therapeutic development of this technique. In order to bypass the temporary dpwnregulation by siRNA, a new generation of shRNA named shRNAmir has developed. Silencing construct with structure similar to microRNA (shRNAmir), mimics a natural microRNA pathway inside the cell. Steroid receptor RNA activator (SRA) is one of the regulators of steroid receptor like ER. Prostate, uterus and breast tissue express a low level of SRA, there is an increase of expression during their tumorgenesis. So SRA may participate in tumorgenesis or proliferation of tumors.
Materials and Methods: We used RNAi technique to silence expression of SRA. The SRA silencer was designed and constructed by Soe-PCR, then cloned into an expression vector pEGFPC1. Human breast cancer (MCF7) cells were transfected with silencer plasmid then the changes in the SRA expression estimated by Real-Time PCR at 24, 72 hours and after 10days.
Result: The results showed about 60% decrease in relative expression of SRA gene, after 72 hours and 10 days, which shows that shRNAmir–SRA could successfully knockdown the expression of target gene.
Conclusion: It seems that the designed shRNAmir may be a suitable tool for a variety of applications because it could stably knockdown the expression of target gene.https://mjms.modares.ac.ir/article_19009_3883cd645fd2fd862058839841a60f97.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Estimation of aflatoxin-albumin adduct in serum sample by optimized HPLC-fluorescenceاندازهگیری میزان آفلاتوکسین متصل به آلبومین در نمونه سرم با استفاده از روش بهینه شده کروماتوگرافی مایع با کارایی بالا و آشکارگر فلورسانس414919010ENعباسصاحبقدملطفیFaribaFarajiDepartment of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranAbdolamir AllamehAllamehDepartment of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranAfshinMohsenifarDepartment of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranBatoolEtemadikiaB.Sc., Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranAliMotaDepartment of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranJournal ArticleObjective: With consideration of lethal effects of aflatoxins specially B1 on human health. Estimation of aflatoxin-albumin adduct, as an important marker of aflatoxin exposure, seems essential. The aim of this study is optimization of HPLC-fluorescence method for measurement of this important marker in blood serum.
Materials and Methods: In this study, blood serum of three groups of rats as A) positive controls (treated with AFB1), B) negative controls (without treatment) and standard rats (treated with radiolabeled AFB1) were used. After albumin isolation using ammunium sulphate and acetic acid, purity of albumin was tested by SDS-PAGE electrophoresis and albumin concentration was quantified by bradford method. Then albumin was hydrolysed by pronase and aflatoxin bound to albumin was released as aflatoxin-lysine. Pronase was precipitated and albumin was digested by aceton in cold, the volume of supernatant was reduced by freeze-drier and injected into HPLC system. Aflatoxin was quantified in comparison to standard rats samples.
Results: The purity of this isolated albumin was confirmed by SDS-PAGE electrophoresis. Albumin concentration in positive, negative and standard samples were 10, 13 and 12.5 mg/ml, respectively. Detection limit (20 pg/mg Alb) for measurement of aflatoxin was determined by HPLC method, specificity and sensitivity of method were 92% and 100% respectively. The mean concentration of AF-Alb adducts in serum of positive control rats was 10 ng/mg Alb and the reproducibility of the method after several repeat was very good.
Conclusion: In this study, for AF-Alb adduct quantification by HPLC method, mobile phase, percentage of solvents and run time were changed and the affinity chromatography before HPLC, was deleted. Therefor HPLC- fluorescence which is a precise and specific method, and since it is fast, highly reproducible and cost effective, also with improvement made, could easily be used for the quantification of this important marker in serum.https://mjms.modares.ac.ir/article_19010_ac55b967f3e3b0edccf5be563e4a6016.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Evaluation of chitosan/poly (vinyl alcohol) nano-composite biocompatibility for neural cells proliferationبررسی زیستسازگاری نانوکامپوزیت کیتوزان/پلیوینیل الکل برای تکثیر سلولهای عصبی516019011ENMohammad AliShokrgozarDepartment of National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, IranFatemehMottaghitalabDepartment of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, IranVahidMottaghitalabDepartment of Textile Engineering, Faculty of Engineering, Guilan University, Rasht, IranMehdiFarrokhiDepartment of National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, IranAliEslamifarDepartment of Clinical Research, Pasteur Institute of Iran, Tehran, IranJournal ArticleObjective: Tissue engineering is an (interdisciplinary field that applies polymeric scaffolds to control tissue formation in three-dinemtion (3D). The scaffold provides the microenvironment (synthetic temporary extracellular matrix) for regenerative cells, supporting cell attachment, proliferation, differentiation, and neo tissue genesis due to their suitable chemical, physical and biological structures. In this study, chitosan/poly (vinyl alcohol) (CS/PVA) was exploited as scaffold for nerve regeneration.
Materials and Methods: Electrospinning was used to fabricate CS/PVA nanocomposites for U373 cells seeding and proliferation. Electrospinning is a versatile and simple method to fabricate non-woven thin layer fibers from polymeric solutions. Consequently, the biocompatibility of CS/PVA nanocomposite was evaluated using biological assays and cell attachment study.
Results: Results indicated that CS/PVA nanocomposites with 15/85 proportion shown an almost homogenous network of the electrospun fibers and confirmed that they can be knitted in meshes and improve U373 cells proliferation and cell attachment.
Conclusion: The nano-sized CS/PVA scaffolds are nontoxic and biocompatible which can promote proliferation of U373 cells and their appropriate adhesion to nanocomposite for improved peripheral nerve regeneration.https://mjms.modares.ac.ir/article_19011_4839e9401ca59a31a153bfc202e3971e.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Identification of methicillin-resistant Staphylococcus aureus by disk diffusion method, determination of MIC and PCR for mecA geneشناسایی استافیلوکوکوس اورئوس مقاوم به متیسیلین با روش انتشار دیسک، تعیین MIC و PCR برای ژن mecA616919012ENShahinNajar PeerayehDepartment of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranAmirAzimianDepartment of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranMohammadMostafaeeDepartment of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranSeyed DavarSiadatDepartment of Bacterial Vaccine and Antigen Preparation, Pasture Institute of Iran, Tehran, IranJournal ArticleObjectives: Staphylococcus aureus is an important cause of serious infection in both hospital and the community. Methicillin-resistant S. aureus (MRSA) is associated with high morbidity and mortality rates with rapid development of resistance. There is a need for early and reliable detection of MRSA infection to direct antibiotic therapy, and more effectively control cross-infection. In this study, resistance to methicillin was detected by a disk diffusion method, the determination of MIC, and the PCR for mecA gene.
Materials and Methods: A total of 174 S.aureus strains were isolated from different clinical specimens from three teaching Hospitals. Antibiotic susceptibility was determined by disk diffusion method, MIC for oxacillin was made by the agar dilution, and mecA gene was identified by specific primers.
Results: The prevalence of MRSA by three methods ranged from 47% to 50%, and mecA positive isolates were more resistant to all of the antibiotic tested than mecA negative isolates. All S. aureus isolates were resistant to penicillin, and susceptible to vancomycin. The results of agar dilution test indicated a low-level resistance to methicillin (MIC>64mg/l). The distribution of MRSA isolates were uniform between three hospitals, and there were not significant differences in the presence of MRSA between isolates from different clinical specimens.
Conclusion: The PCR method was the best test for routine detection of MRSA in the present study. An additional benefit of the mecA PCR is the potential to generate a susceptibility report, 24h earlier than the time of generation of results of conventional susceptibility testing methods.https://mjms.modares.ac.ir/article_19012_e35026a2b91f352db22bfef0e693c2e3.pdfTarbiat Modares UniversityPathobiology Research2538-300012320090901Antifungal effect of green tea leaf (Camellia sinensis) polyphenols on Candida albicansاثر ضد قارچی پلیفنلهای برگ سبز چای (Camellia sinensis) بر کاندیدا آلبیکنس717719013ENMohammad HosseinYadegariDepartment of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranZahraNasrollahiDepartment of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranSeyed MohammadMoazeniDepartment of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranJournal ArticleObjective: In this study the susceptibility of Candida albicans to inhibitory effect of polyphenols under varying time (24 and 48 hours) conditions were evaluated.
Materials and Methods: Green tea leaf polyphenols were extracted and analyzed by chromatography. Among polyphenols, Catechin showed stronger antifungal activity against C. albicans PTCC-5027. Catechin's MIC90 (The concentration of Catechin causing 90% growth inhibition of tested strain of C. albicans) and MFC (The minimum antifungal susceptibility of Catechin) were determined by Macro dilution test and calculation after 24 and 48 hours.
Results: The antifungal activity of Catechin was time dependent. Catechin's MIC for 0.5×10³, 1×10³ and 2×10³ cells/ml was 12.5, 25 and 100 mg/ml after 24h respectively. The results after 48h for 0.5×10³, 1×10³ and 2×10³ cells/ml were 6.25, 12.5 and 25 mg/ml respectively. Fluconazol was tested on C. albicans PTCC-5027 and the results indicated that this strain of Candida is fluconazol resistant. Data shown are from three separate experiments and were analysed statistically.
Conclusion: C. albicans PTCC-5027 is fluconazol resistant, however green tea leaf polyphenols especially Catechin could inhibit the growth of this yeast at MIC and MFC concentrations. On the basis of the obtained results the green tea leaf contains effective antifungal components. Since the common and generic antifungal drugs possess some side effects and also there is an increasing drug resistance, it is hoped that consumption of herbal drugs may help to cure fungal diseases and to avoid the side effects of antimycotics as a good replacement.https://mjms.modares.ac.ir/article_19013_edd6a7e04e7f4dd932a5c44f282f9b4e.pdf