Tarbiat Modares UniversityPathobiology Research2538-300015220120601The Effect of Artemether-induced Apoptosis in Promastigotes of Leishmania major (MRHO/IR/75/ER) under In-vitro Conditionsبررسی اثر داروی آرتمتر بر مرگ سلولی پروماستیگوتهای سویه ایرانی (MRHO/IR/75/ER) انگل لیشمانیا ماژور در شرایط آزمایشگاهی11019132ENParisaEbrahimisadrM.Sc, Department of Parasitilogy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranFatemehGhaffarifarAssociated Professor, Department of Parasitilogy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranZuhair MohammadHassanProfessor, Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranNasibehBeheshtiM.Sc, Department of Parasitilogy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranJournal Article19700101Objective: Leishmaniasis is one of the significant causes of morbidity and mortality in several countries. It is an important problem in endemic areas such as Iran. The goal in treatment of leishmaniasis is to reduce the disease period and leave no evidence of any remaining scars or lesions. A derivative of artemisinin is artemether. Scientists believe that the strong action of artemether against parasites is due to the presence of an endoperoxide bridge. Due to problems in the treatment of <em>Leishmania</em><em> major</em>, in this research we have studied the effect of artemether on <em>Leishmania</em><em> major</em> under in vitro conditions. Methods: Parasites were cultured in NNN and RPMI, after which artemether at concentrations of 5, 10, 25, 50 and 100 μg/ml were used for the promastigote assay. Apoptosis was detected by flow cytometry and DNA ladder assay. Results: The inhibitory concentration (IC50) of artemether was determined to be 25 μg/ml. The percentage of apoptotic promastigotes at 25 μg/ml of artemether was 42.28. The results of DNA fragmentation show that exposure of <em>Leishmania</em><em> major</em> promastigote cells to 25 μg/ml of artemether lead to DNA fragmentation. Conclusion: We have proven the effect of artemether on apoptosis of <em>Leishmania</em><em> major</em> by flow cytometry and the DNA ladder assay.https://mjms.modares.ac.ir/article_19132_75b769beb7e88dc7c59c1adb2006533c.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Preparation of Methylene Blue-containing Nanoliposomes and Determination of Stability, Biological Distribution and Drug Release after Sonication by 1 MHz Ultrasound Wavesتهیه نانولیپوزوم پلیمری حاوی داروی حساس کننده نوری متیلن بلو و ارزیابی پایداری، توزیع زیستشناختی و رهاسازی دارو با تابش امواج فراصوتی یک مگاهرتز112219133ENAliEbrahiminiaPh.D. Candidate, Department of Medical Physics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranManijheMokhtari-DizajiProfessor, Department of Medical Physics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranTayebehToliyatAssociated Professor, Department of Pharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, IranJournal Article19700101Objective: In order to overcome the limitation of systemic administration of methylene blue, this study investigated the encapsulation of methylene blue in polymeric liposomes and drug release following sonication. Methods: We encapsulated methylene blue into nanoliposomes. The dynamic light scattering (DLS) method was used to measure the size distribution of the liposomes. After loading methylene blue into these liposomes, both drug encapsulation efficiency and stability were fluorometrically determined. Biodistribution of drug was studied in vivo in a mouse model of adenocarcinoma tumor cells. The amount of drug released upon 1 MHz sonication at an intensity of 2 W/cm<sup>2</sup> was fluorometrically verified in vitro. Results: DLS studies showed that the synthesized liposomes had an average size of 66.19±4.49 nm. Methylene blue was efficiently encapsulated in nanoparticles at an average of 65.21±3.47%. Stability of the generated liposomes decreased with time. Biodistribution study revealed that the drug content in the group that received liposomal drugs in their tumor tissue was significantly higher than in the group that received methylene blue in its free form and in the heart was inverse (PConclusion: This study has shown that fabricated liposomes are suitable for the encapsulation and delivery of hydrophilic photosensitizers such as methylene blue. Ultrasound-triggered release was achieved by the use of a 1 MHz ultrasound.https://mjms.modares.ac.ir/article_19133_baaf111c2bdc129a356a14f79fefd224.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Remyelination of Demyelinated Rat Spinal Cord Model by Transplanting Neural Stem Cellsترمیم میلین با استفاده از پیوند سلولهای بنیادی عصبی در نخاع موش صحرایی مدل تخریب غلاف میلین233419134ENSamaneAdibM.Sc. Student, Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranTakiTirahiProfessor, Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranTaherTaheriNeurosurgeon, Shefa Neuroscience Research Center, Khatam al-Anbia Hospital, Tehran, IranJournal Article19700101Objective: Demyelination of CNS axons occurs under pathological conditions such as multiple sclerosis and spinal cord injuries, but can be repaired by cell therapy. Within the CNS remyelination can be achieved by transplantation of neural stem cells (NSCs). NSCs are self-renewing cells that maintain the capacity to differentiate into CNS-specific cell types and can differentiate into the three main neural phenotypes: astroglia, oligodendroglia and neurons. They may also replace or repair diseased CNS tissue. Methods: Bone marrow stromal cells (BMSCs) were aseptically isolated from the tibia and femurs of young adult Sprague Dawley rats. BMSCs were evaluated by fibronectin and CD31 markers. BMSC-derived NSCs were evaluated by nestin and NF-68. An ethidium bromide-induced demyelinated dorsal column lesion was produced in young adult rats. Transplanting NSCs derived-BMSCs into demyelinated lesions after 3 days in adult rat spinal cords was done. Three weeks after transplantation of NSCs, the spinal cords were processed to evaluate remyelination by Luxol fast blue staining. Results: After passage 3, BMSCs were evaluated and the result, showed the percentage of immunoreactive cells to fibronectin (94.7±2.65), however BMSC-derived NSCs expressed nestin (86.15±0.64) and NF-68 (84.55±0.94) which correlated with fibronectin down regulation. Histologically, the lesions showed slightly irregular elongated areas and had an average length of 1336.36±39.43 µm. Transplanted NSCs were capable of eliciting remyelination. Conclusion: These data support the conclusion that transplantation of NSCs results in functional remyelination of a dorsal column lesion and have valuable applications in the treatment of neurodegenerative diseases such as spinal cord injuries.https://mjms.modares.ac.ir/article_19134_f3c1eaec9dce37a8f6b301a80570dff9.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Comparison of Rotavirus RF Strain and HSV-1 Titration by CCID50% and Plaque Assaysمقایسه تیتراسیون روتاویروس گاوی سویه RF و ویروس هرپس سیمپلکس نوع 1 با دو روش پلاک و CCID50%354519135ENAliTeimooriPh.D. Candidate, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranHooriehSoleimanjahiAssociated Professor, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranFarzanehPourasgariAssistant Professor, Department of Biotechnology, Razi Vaccine and Serum Research Institute, Karaj, IranJournal Article19700101Objective: Titration of viruses is important to determine the quantity of virus in vaccine development, master virus seed stock preparation, viral vector studies and virus replication. In this study, we compared the CCID50% and plaque assay as a standard titration method for rotavirus (RF) and HSV-1. Methods: The MA104 and Vero cells were inoculated by RF and HSV-1 in 6- and 96-well plates. Following infection and adsorption, the optimal time for the cytopathic effect caused by the viruses was noted and the results compared. Results: The CPE (Cytopathic Effect) of RF was observed in less than 18 hours, which increased until 72 hours after inoculation. In HSV-1, the CPE was observed 24 and 72 hours after inoculation. The virus titration in the plaque assay was monitored at 96 hours post-infection for RF and at 72 hours post-infection for HSV-1. In both viruses the plaque titer method was lower than the CCID50 method, since the results indicated that 1 CCID50% was equal to 0.7 PFU. Conclusion: The plaque assay is one of the most accurate methods for viral titration. For the plaque assay, individual lesions may be isolated, which the plaques can be counted. The CCID50% method is not applicable for purification of homogenous viruses, nor is this technique reproducible.https://mjms.modares.ac.ir/article_19135_73475aa6f2950924842dd59077a7be8e.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Fabrication and Characterization of Regenerated Silk/bioglass Composites for Bone Tissue Engineeringساخت و بررسی خواص زیستی داربستهای کامپوزیتی ابریشم بازیابی شده / شیشه زیستفعال برای مهندسی بافت استخوان476019136ENSahbaMobiniAssistant Professor, Reproductive Biotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, IranMehranSolati-HashjinAssociated Professor, Department of Biomaterial, Faculty of Medical Engineering, Amirkabir University of Technology, Tehran, IranSaeedHesarakiAssistant Professor, Material and Energy Research Center, Karaj, IranMichaelGelinskyProfessor, Centre for Translational Bone, Joint and Soft Tissue Research, University Hospital Carl Gustav Carus and Medical Faculty of Technische Universität Dresden, GermanyJournal Article19700101Objective: One of the major issues in bone tissue engineering is the design and fabrication of bioactive, bioresorbable porous 3D scaffolds capable of maintaining their structure and integrity over a predictable period of time. One such approach is the fabrication of composite scaffolds. Methods: In this study we present fabrication and characterization of novel silk/bioglass-composite scaffolds. Regenerated fibroin was constructed from mulberry silk cocoons and calcium silicophosphate bioactive glass was made by sol-gel processing. For fabrication of a homogenous composite, grained bioglass particles were modified with 3-aminopropyltriethoxysilane coating. Fibroin/bioglass composite scaffolds were fabricated by the freeze-dry technique at different concentrations. Results: Silk protein extract was evaluated by FTIR and XRD methods. FTIR spectrum showed sharp amide peaks at 1655 cm<sup>-1</sup> and 1530 cm<sup>-1</sup> wave lengths, which confirmed the existence of fibroin. XPS analysis demonstrated that the amino groups were established on the surface of the glass powder. The fabricated 3D scaffolds were morphologically analyzed by scanning electron microscopy, which showed uniformly dispersed bioglass particles in all structures. Scaffolds were seeded with human mesenchymal stem cells for 21 days. Conclusion: Considering the cytocompatibility of the scaffolds and osteogenic differentiation during three weeks, it could be concluded that the appropriate combination of structural and biological properties make the silk/bioglass composite scaffold a probable choice for potential use in bone tissue engineering.https://mjms.modares.ac.ir/article_19136_2cfd22322a832d773c4218f4559fa174.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601A Suicide-inducing Vector Carrying the Oct-4 Promoter for Eradication of the AGS Cancer Cell Lineناقل حاوی پروموتور Oct4 برای القای مرگ سلولی به رده سلول سرطانی AGS617219137ENRoshanakNajafiPh.D. Candidate, Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranMajidSadeghizadehProfessor, Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranMajidSadeghizadehProfessor, Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranSeyed JavadMowlaMolecular Genetics, Faculty of biological sciences, Tarbiat Modares UniversityJournal Article19700101Objective: In an attempt to develop safer and more effective gene therapy approaches as a realistic treatment for various forms of cancer, researchers are increasingly using tumor-specific promoters (TSP) to drive the expression of the gene of interest<em> </em>and eradicate cancer cells. In this study, for the first time we introduce the Oct-4 promoter as a cancer-specific promoter with a high efficacy. Methods: We cloned Oct-4 promoter and enhancers into a pGl3 control reporter vector and analyzed the expression of luciferase as a reporter gene in an AGS gastric cell line. Next, we used a suicide-inducing vector that included an Oct-4 promoter and the TK gene in the presence of the non-toxic prodrug, ganciclovir, to eradicate cancer cells. Cells were treated with PI and connexin V. FACS analysis was conducted to assess the effect of the system on cell cycle and apoptosis induction. Results: Under the activity of the Oct-4 promoter, luciferase expression was three-fold higher than the SV40 promoter. The HSV/TK/GCV system activated by the Oct-4 promoter and enhancers induced apoptosis (86.17%) in the AGS cell line. We verified that this system induced S-phase/G2-phase cell cycle arrest in the AGS cell line. Conclusion: These data indicate that the Oct-4 promoter is active in the AGS cell line. Oct-4 gene is expressed in a wide variety of tumors but not in normal cells. Thus, Oct-4 appears to be a promising tumor-specific promoter for numerous tumors.https://mjms.modares.ac.ir/article_19137_cd483cdcbfa62470fa1ec3771c5fe370.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Evaluation of the Effects of Interferon on Stability of Control Genes in Huh-7 and HepG2 Cellsبررسی تأثیر اینترفرون بر پایداری ژنهای کنترل در ردههای سلولی Huh-7 و HepG2738519138ENAtiehHashemiPh.D. Candidate, Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, IranFarzinRoohvandAssistant Professor, Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, IranMohammad-hosseinGhahremaniPharmacology and toxicology department, Pharmacology School, University of Tehran, Tehran, IranRositaEdalatM.Sc., Department of Virology, Pasteur Institute of Iran, Tehran, IranJournal Article19700101Objective: Understanding gene expression variations by using RNA transcript analysis methods during hepatic viral protein interactions with the IFN pathway in hepatic cell lines has recently gained importance. One of the most powerful techniques in gene expression quantification is quantitative real-time RT-PCR. Reference genes used as normalizer in this method may be affected across various experimental conditions or treatments. Hence, in the present study, the influence of IFN-a treatment on the mRNA levels of common reference genes including ACTB, GAPDH, TBP, HPRT1 and HMBS was evaluated in Huh-7 or HepG2 cell lines. Methods: Cells were treated with different concentrations of IFN-a. Then, using geNorm and NormFinder programs, we evaluated the expression stabilities of the above prominent reference genes in three sample groups that included each hepatic cell line and the total data sets. Results: HPRT-1 and GAPDH were the most stable reference genes in the Huh-7 cell line, whereas ATCB, HMBS and GAPDH were the most stable in the HepG2 cell line. TBP was one of the least stable reference genes in the three studied groups. Conclusion: This investigation will provide appropriate reference genes for standardization of quantitative real-time PCR data in an IFN-a stimulated model of hepatocyte cell lines.https://mjms.modares.ac.ir/article_19138_e0c5b770acef5dc9a9c462548446c082.pdfTarbiat Modares UniversityPathobiology Research2538-300015220120601Rapid Detection of Trichophyton rubrum in Clinical Samples from Tinea Unguium using PCRتشخیص سریع و حساس اونیکومایکوزیس در نمونههای بالینی مشکوک به درماتوفیتوزیس ناخن (ترایکوفیتون روبروم) با استفاده از روش PCR879519139ENSeyed AliMoallemzadehPh.D. Candidate, Department of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranMohammad HosseinYadegariAssociated Professor, Department of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranParvinMansouriProfessor, Department of Dermatology, Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, IranMasumehRajabi BazlAssistant Professor, Department of Clinical Biochemistry, Faculty of Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, IranRezaKachueiAssistant Professor, Molecular Biology Research Center, Baqiyatallah University of Medical Science, Tehran, IranJournal Article19700101Objective: Dermatophytosis is one of the most common pandemic fungal infections that is a major health problem in cities and villages. This study aims to evaluate PCR sensitivity and accuracy in the detection of nail dermatophytosis compared to conventional direct and culture detection methods, and performs an assessment of <em>Trichophyton rubrum</em> in patients suspected of having nail dermatophytosis.
Methods: This experiment was a descriptive-experimental study carried out on 71 nail samples obtained from patients with suspected nail dermatophytosis. All clinical samples of nails or chips were divided into three sections and each section underwent direct examination, culture and molecular tests. In the molecular test, we used fungal rRNA universal primers (ITS1 and ITS4) and <em>Trichophyton rubrum</em>-specific primers.
Results: In this study, for the first time in Iran and based on a modified protocol, DNA was directly extracted from tissues of infected nails in less than five hours. Additionally a comparison of the results obtained from routine laboratory methods such as direct examination and culture with PCR verified the high sensitivity and accuracy of PCR compared to the other studied methods.
Conclusion: PCR, as a rapid, accurate method, can be a good replacement for conventional culture and direct examination.https://mjms.modares.ac.ir/article_19139_d5aa64e4dbf9684c9d269395bc3de672.pdf