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دکتر مجید تفریحی

  • Fatemeh Eizadifard, Majid Tafrihi
    Background

    Ferula gummosa Boiss. is a widely recognized species native to Iran, primarily found in the northern and northeastern regions. It has long been valued for its medicinal properties, particularly in traditional medicine, where its gum is used to treat various gastrointestinal disorders and infections.

    Objectives

    However, despite its established benefits, there is a significant research gap regarding its potential anti-cancer properties, specifically against skin cancer cells.

    Methods

    This study aimed to assess the cytotoxic and inhibitory effects of the gum of F. gummosa on the A-375 cell line using several assays, including MTT, clonogenic, acridine orange/ethidium bromide (AO/EtBr) staining, DNA degradation, and in vitro wound-healing experiments.

    Results

    The findings revealed that the gum exhibited notable cytotoxicity on A-375 cells, achieving a 50% inhibitory concentration (IC 50 ) at 8 µg/mL after 48 hours of treatment (compared to control cells) (P < 0.01). The observed DNA degradation pattern suggested a reduction in cell viability, likely due to apoptosis induction. Microscopic examinations showed nuclear condensation and a significant suppression of colony formation in A-375 cells treated with 8 and 10 μg/mL concentrations of the gum, compared to untreated cells. Additionally, wound-healing assessments demonstrated the gum’s ability to inhibit cell migration in contrast to the untreated group.

    Conclusions

    These findings suggest that F. gummosa exhibits significant inhibitory effects on melanoma cancer cells, making it a promising candidate for further investigation.

    Keywords: Ferula Gummosa, A-375 Cells, Cytotoxicity, Migration, Cell Proliferation, Melanoma Cancer, Apoptosis
  • Ghazaleh Malekizadeh, Omid Jazayeri*, Morteza Alijanpour, Majid Tafrihi
    Background

    Methylmalonic acidemia (MMA) is a rare autosomal recessive metabolic disorder resulting from a genetic defect in methylmalonyl-CoA mutase (MCM) or a defect in the biosynthesis of its cofactor, adenosyl-cobalamin (AdoCbl). The disease is caused by a mutation in six main genes (MUT, MMAA, MMAB, MMADHC, MMACHC, and MCEE). In this investigation, we estimate MMAdisease gene frequencies globally and report MMA-causative mutations in the Iranian population.

    Methods

    Human gene mutation database (HGMD) has been utilized to estimate MMA-disease gene frequencies. To compile MMA mutations in Iran, we systematically reviewed PubMed, Google Scholar, CIVILICA, Magiran, and SID databases to explore relevant articles in English and Persian.

    Results

    The frequencies of causative genes among MMA patients at the global level were as follows: MUT (64.14%), MMACHC (17.74%), MMAA (13.48%), MMAB (7.1%), MMADHC (2.9%), and MCEE (0.85%). Until February 11, 2024, 24 MMA mutations had been compiled from the Iranian population; of which 11 mutations (45.8%) had been diagnosed only in Iran and had not been addressed in other populations yet.

    Conclusion

    Collection and recognition of MMA mutations in the Iranian population can be helpful for early diagnosis and treatment before the onset of neurological manifestations in neonates.

    Keywords: Methylmalonic Academia, Mutation, Iranian Population, Methylmalonyl-Coa Mutase
  • Fatemeh Eizadifard, Majid Tafrihi *, Maryam Mohadjerani
    Objective
    Ferula gummosa Boiss is a well-known Iranian endemic plant that has been used in Iranian traditional medicine against various diseases. This study aimed to evaluate the antioxidant and cytotoxic capacity of F. gummosa gum on prostate cancer PC-3 cells.
    Materials and Methods
    In this study, we evaluated the total phenolic and flavonoid contents, and antioxidant potentials of the gum. The MTT experiment was conducted to assess the cytotoxic potential of the gum on PC-3 cells. The clonogenic, micronucleus formation, and acridine orange/ethidium bromide staining methods were used to evaluate the survival and proliferation of PC-3 cells. DNA degradation and caspase 3/7 activity evaluations were used to assess apoptosis. The inhibitory effect on the migration of PC-3 cells was examined by in vitro wound-healing experiment.
    Results
    Total phenolic and flavonoid contents, and antioxidant potential of the gum were 9.22 mg of gallic acid equivalent (GAE)/g, 3.6 mg of quercetin equivalents (QE) /g of the extract, and 13 μg/ml, respectively (compared to gallic acid and quercetin, respectively) (p<0.05). The IC50 value was 9.14 µg/ml for 48 hours (compared to non-treated cells) (p<0.01).  The pattern of DNA degradation, and caspase 3/7 activity levels (compared to non-treated cells) (p<0.05) proposed decreased cell viability that may be due to apoptosis induction. Microscopic observations revealed nuclear condensation, a significant increase in the formation of micronuclei, and inhibition of forming colonies (compared to non-treated cells) (p<0.01) in PC-3 cells treated with 8 and 10 μg/ml of the gum. Wound-healing assessment showed the migration suppression potentials of the gum (compared to non-treated cells) (p<0.05).
    Conclusion
    These results indicate that F. gummosa has considerable antioxidant and cytotoxic properties that can make it a good nominee for subsequent investigations.
    Keywords: Ferula gummosa, PC-3 cells, Cytotoxicity, Antioxidant potential, Caspase-3, 7 activity
  • Parvaneh Hajipour, Fatemeh Eizadifard, Majid Tafrihi*
    Background

    Teucrium persicum, a well-known Iranian endemic plant, has been used to treat various diseases in Iranian traditional medicine.

    Objectives

    This study aimed to assess the antioxidant and cytotoxic effects of chloroform and ethyl acetate extracts of T. persicum on cancer cell lines.

    Methods

    The total phenolic and flavonoid contents, as well as the antioxidant potential of chloroform and ethyl acetate extracts were assayed. The GC-MS analysis was used to detect the chemical constituents of these two extracts. The cytotoxicity effects of extracts on two cancer cell lines, PC-3, SW-480, and HEK-293, as normal cell lines were evaluated using the MTT assay.

    Results

    The average total phenolic contents of chloroform and ethyl acetate extracts were found to be 48 ± 0.2 and 25 ± 0.4, respectively (P < 0.05). The total flavonoid contents of chloroform and ethyl acetate extracts were measured as 34 ± 0.6 and 36 ± 0.6 mg EQ/g of the dried extract, respectively (P < 0.05). The antioxidant potential of chloroform and ethyl acetate extracts were calculated as 2.5 ± 0.013 and 3 ± 0.0023 µg/mL, respectively (P < 0.05). According to the GC-MS analysis, 60 and 61 different bioactive compounds were detected in chloroform and ethyl acetate extracts, respectively. MTT results showed that the SW-480 cell line was more sensitive than PC-3 and HEK-293 cells.

    Conclusions

    It was concluded that T. persicum had significant antioxidant and cytotoxic properties. Taking our study results into account, cancer inhibiting properties of T. persicum were strongly supported. However, it was recommended that further specific and detailed biochemical and molecular studies should be conducted in order to discover the effective compound(s) and mechanism(s) responsible for producing these effects.

    Keywords: Teucrium persicum, Chloroform Extract, Ethyl Acetate Extract, Antioxidant Potential, Cytotoxicity
  • Afshin Samimi Nemati, Majid Tafrihi*, Fatemeh Sheikhi, Abolfazl Rostamian Tabari, Amirhossein Haditabar
    Background

    Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has currently caused a significant pandemic among worldwide populations. The high transmission and mortality rates of the disease necessitate studies for rapid designing and effective vaccine production. This study aims to predict and design a novel multi-epitope vaccine against the SARS-CoV-2 virus using bioinformatics approaches. 

    Materials and Methods

    Coronavirus envelope proteins, Open Reading Frame 7b (ORF7b), Open Reading Frame 8 (ORF8), Open Reading Frame 10 (ORF10), and Nonstructural protein 9 (Nsp9) were selected as targets for epitope mapping using Immune Epitope Data Bank (IEDB) and BepiPred 2.0 servers. Also, molecular docking studies were performed to determine the candidate vaccine’s affinity to Toll-Like Receptor (TLR3, TLR4) and Major Histocompatibility Complex (MHC I and MHC II) molecules. Thirteen epitopes were selected to construct the multi-epitope vaccine. 

    Results

    We found that the constructed peptide has valuable antigenicity, stability, and appropriate half-life. The Ramachandran and ERRAT plots approved the quality of the predicted model after the refinement process. Molecular docking investigations revealed that antibody-mode in the ClusPro 2.0 server showed the lowest binding energy for MHC I, MHC II, TLR3, and TLR4.

    Conclusion

    The designed vaccine has a good antigenicity and stability and could be a proper vaccine candidate against the Coronavirus Disease 2019 (COVID-19) infectious disease though, in vitro and in vivo experiments are necessary to complete and confirm our results.

    Keywords: COVID 19, SARS-COV-2, Vaccine, ORF, Immunoinformatics
  • Anahita Naeimi, Majid Tafrihi *, Maryam Mohadjerani
    Objective
    Teucrium persicum is an Iranian endemic plant used in Iranian traditional medicine.
    Materials and Methods
    The total phenolic and total flavonoid contents, and antioxidant potential of the methanolic extract of T. persicum were determined. The MTT test was used to evaluate the inhibitory effect of the extract on the viability of A-375 cells. The clonogenic, micronucleus formation, and acridine orange/ethidium bromide staining methods were used to evaluate the survival and proliferation of A-375 cells. Apoptosis was evaluated by using DNA fragmentation assay and measuring the activity of caspase 3/7. To study the effect of the extract on the migration of A-375 cells, the in vitro wound-healing (scratch) assay was employed.
    Results
    The average total phenolic and flavonoid contents and antioxidant properties of the extract were 6.97±0.011 mg Ellagic acid (EGA)/g, 46.83±0.0019 mg of the ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline; EQ)/g of dried extract, and 10±0.002 μg/ml, respectively. The IC50 value of the T. persicum methanolic extract was 13 μg/ml for 48 hr. The DNA fragmentation pattern and the activity of caspase3/7 suggested that the reduction of the cell viability may be due to apoptosis induction. Microscopic observations showed nuclear condensation, a considerable increase in micronuclei formation, and inhibition of the colony formation in A-375 cells treated with 7 μg/ml to 15 μg/ml of the extract. Wound-healing assay supported the anti-migration activity of the extract.
    Conclusion
    T. persicum has significant antioxidant and cytotoxic properties. Surely, more detailed molecular and biochemical studies are needed to find the mechanism(s) behind these effects.
    Keywords: Teucrium persicum, A-375 cells, Antioxidant potential, Cytotoxicity, Caspase 3, 7, Genotoxicity
  • Hamid Reza Sharifzadeh, Majid Tafrihi *, Nouredin Moradi, Naghmeh Gholipour
    Chromosomal aneuploidies are the most chromosomal abnormalities at birth due to maternal meiosis I errors. Pregnancies with autosomal chromosomal aneuploidies that survive are namely trisomies 13 (Patau syndrome), 18 (Edward syndrome), and 21 (Down syndrome), account for 89% of chromosome abnormalities. Quantitative fluorescent polymerase chain reaction (QF-PCR) which amplifies specific DNA sequences called short tandem repeats (STRs), by using fluorescently labeled primers is a rapid technique for prenatal diagnosis of common aneuploidies. In this study, DNA extraction was performed from 100 samples isolated from muscle tissue of aborted fetuses. The analysis was performed by multiplex QF-PCR using a panel of 25 STRs markers for chromosomes X, Y, 13, 18, and 21. Our results showed that 20% of abortions were due to aneuploidy. 53% of mothers who had abortions were aged 26-35 years old and 32% of them were aged 36-45 years old. The analysis of muscle samples of aborted fetuses indicated that 20 samples showed chromosomal aneuploidy. Of the abnormal cases, 10 cases (~50 %) showed trisomy 21 followed by trisomy 18 (7 cases, ~35%), Klinefelter syndrome (2 cases, ~10 %), and showed trisomy X (1 case, ~5 %). Our results indicated that the D21S1414 marker showed the highest rate of heterozygosity in the study population. Besides some limitations of this study such as sample size, these results suggest that one of the causes of these abortions could be maternal age. We concluded that QF-PCR could be a rapid and reliable method to screen prenatal chromosomal aneuploidy and allow appropriate counseling.
    Keywords: Abortion, Chromosomal aneuploidy, Fetus, Iranian population, QF-PCR
  • Majid Tafrihi *, Sogand Kalantari, Mohammad Shokrzadeh

    E-cadherin is a tumor suppressor protein that plays a crucial role in cell-cell adherens junction and tissue architecture and it is hypothesized to participate in carcinogenesis. It has been shown that a polymorphism in the upstream of the transcription start site of the CDH1 gene affects E-cadherin transcriptional regulation and seems to be associated with a variety of cancers. For the first time, we investigated the association of the rs16260 in the 5'-untranslated region of the CDH1 gene with gastric cancer in Iranian population. Seventy- eight patients with gastric cancer and 72 healthy individuals were included and genotyped for this SNP using the PCR-RFLP method. Our results showed that the frequency of the AA genotype in gastric cancer patients (16 of 78, 20.5%) was higher than healthy individuals (9 of 72, 12.5%), the frequency of the A allele in the patients group was higher than controls (OR=1.231, 95% CI= 0.772-1.962, p-value= 0.383), but statistical analysis revealed the absence of association between AA genotype and gastric cancer risk (OR=1.719, 95% CI= 0.656-4.502, p-value= 0.268). In conclusion, our results suggest that this substitution and the AA genotype have not a major impact on the individual’s susceptibility to gastric cancer, and therefore this SNP may be an ethnicity-dependent risk factor. Further works with larger sample size and including other criteria such as H. pylori infection status are needed for more accuracy.

    Keywords: Gastric cancer, CDH1, E-cadherin protein, promoter, Single nucleotide polymorphism
  • Abasalt Hossienzadeh, Colagar*, Mohammad Javad Haghighatnia, Zahra Amiri, Maryam Mohadjerani, Majid Tafrihi
    Microsatellites or simple sequence repeats (SSRs) are very effective molecular markers in population genetics, genome mapping, taxonomic study and other large-scale studies. Variation in number of tandem repeats within microsatellite refers to simple sequence length polymorphism (SSLP); but there are a few studies that are showed SSRs replication slippage may be occurred during in vitro amplification which are produced ‘stutter products’ differing in length from the main products. The purpose of this study is introducing a reliable method to realize SSRs replication slippage. At first, three unique primers designed to amplify SSRs loci in the great gerbil (Rhombomys opimus) by PCR. Crush and soak method used to isolate interesting DNA bands from polyacrylamide gel. PCR products analyzed using by sequencing methods. Our study has been shown that Taq DNA polymerase slipped during microsatellite in vitro amplification which led to insertion or deletion of repeats in sense or antisense DNA strands. It is produced amplified fragments with various lengths in gel electrophoresis showed as ‘stutter bands’. Thus, in population studies by SSRs markers recommend that replication slippage effects and stutter bands have been considered.
    Keywords: Microsatellites, Taq polymerase slippage, Replication slippage
  • مجید تفریحی*، روح الله نخعی سیستانی
    پروتئین E-cadherin یکی از اعضاء فوق خانواده پروتئین های E-cadherin است که بطور عمده در سطح سلول های بافت های اپیتلیال بیان می شود. کاهش بیان این پروتئین علاوه بر تضعیف اتصالات بین سلولی، موجب بروز پدیده Epithelial-mesenchymal transition (EMT) و مهاجرت و متاستاز سلول های سرطانی بافت-های اپیتلیال می شود. در طب سنتی، از انار (Punica granatum) برای درمان بسیاری از بیماری ها از جمله اسهال، بیماری های قلبی و فشار خون استفاده می شود. در این تحقیق اثر عصاره استونی میوه انار بر میزان بیان پروتئین های β-catenin و E-cadherin در سلول های PC-3 بررسی شد. نتایج آزمایشات MTT نشان داد که عصاره استونی میوه انار قادر است حیات سلول های PC-3 را بخصوص در غلظت های بالاتر از μg/ml 100 مهار کند. همچنین IC50 عصاره استونی دانه انار در حدود μg/ml 86 تخمین زده شد. نتایج آزمایشات وسترن بلاتینگ نیز نشان داد که تیمار سلول های PC-3 با غلظت های μg/ml 20 و μg/ml50 عصاره میوه انار موجب افزایش مقدار پروتئین β-catenin و E-cadherin به میزان تقریبی 5/1 و 2/2 برابر می شود. نتایج این پژوهش پیشنهاد می کند احتمالا میوه انار پتانسیل بالایی در مهار متاستاز سلول های سرطانی دارد.
    کلید واژگان: انار، سلولهای PC، 3، پروتئین β، catenin، پروتئین E، cadherin
    Rohollah Nakhaei Sistani, Majid Tafrihi*
    E-cadherin protein is a member of cadherin superfamily which expressed mainly at the surface of epithelial cells. Reduced expression of this protein results in cell adhesion weakening, EMT (Epithelial-mesenchymal transition), migration and metastasis of epithelial cancer cells. In traditional medicine, pomegranate (Punica granatum) is used to treat several diseases including diarrhea, heart diseases and blood pressure disorders. In this study, we examined the effect of pomegranate acetone extract on the expression of β-catenin and E-cadherin proteins in PC-3 cancer cells. The results of MTT experiments showed that acetone-derived extract of pomegranate inhibits viability of PC-3 cells, especially in concentrations higher than100 μg/ml. The IC50 value of pomegranate extract was estimated to be 86μg/ml. The results of western blotting experiments showed that concentrations of 20 and 50 μg/ml of pomegranate extract led to 1.5- and 2.2-folds increase in β-catenin and E-cadherin protein levels, respectively. The results of this study suggest that pomegranate fruit may have great potentials in inhibition of cancer cell metastasis.
    Keywords: Pomegranate, PC, 3 cells, β catenin protein, E, cadherin protein
فهرست مطالب این نویسنده: 10 عنوان
  • دکتر مجید تفریحی
    دکتر مجید تفریحی
    استادیار زیست شناسی سلولی و مولکولی- دانشکده علوم، دانشگاه مازندران، بابلسر، ایران
نویسندگان همکار
  • دکتر مریم مهاجرانی
    دکتر مریم مهاجرانی
    دانشیار بیوشیمی، زیست شناسی سلولی و مولکولی، دانشکده علوم پایه، دانشگاه مازندران، بابلسر، ایران
  • فاطمه شیخی
    فاطمه شیخی
    دانش آموخته ارشد بیوتکنولوژی مولکولی، دانشگاه صنعتی مالک اشتر، تهران، ایران
  • دکتر امید جزایری
    دکتر امید جزایری
    استادیار زیست شناسی سلولی و مولکولی، دانشگاه مازندران، بابلسر، ایران
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