Publications by authors named "P Farrokh"

18 Publications

Effect of Mouse Ovarian Vitrification on Promoter Methylation of Inhba and Inhbb in Granulosa Cells of Follicles.

Cryo Letters 2021 Mar-Apr;42(2):67-72

School of Biology, Damghan University, Damghan; Institute of Biological Sciences, Damghan University, Damghan, Iran.

Background: Cryopreservation can induce cellular, genomic, and epigenetic abnormalities.

Objective: To analyse the impact of ovarian vitrification on follicular development and its epigenetic effect on promoter methylation of Inhba and Inhbb in granulosa cells.

Materials And Methods: Mouse ovaries were divided into control, toxicity, and vitrified groups. The growth and development of follicles were examined. After in vitro culture of follicles, DNA was extracted from isolated granulosa cells and treated with sodium bisulfite. The promoter methylation of Inhba and Inhbb was analyzed by direct PCR sequencing.

Results: Vitrification reduced the growth of follicles; however, antral cavity formation was not influenced negatively. Vitrification reduced the percentage of 5-methylcytosine in the Inhba promoter, while CpG sites in the promoter of Inhbb remained unmethylated.

Conclusion: Vitrification had adverse effects on follicle growth and the epigenetics of granulosa cells. The results of the current study show that vitrification methods of ovary need more improvement.
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May 2021

Epigenetic effects of Bisphenol A on granulosa cells of mouse follicles during in vitro culture: An experimental study.

Int J Reprod Biomed 2021 Feb 21;19(2):129-136. Epub 2021 Feb 21.

School of Biology, Damghan University, Damghan, Iran.

Background: Bisphenol A (BPA), a synthetic endocrine-disrupting chemical, is a reproductive toxicant. Granulosa cells have significant roles in follicle development, and KIT ligand (KITL) and Anti-Müllerian hormone (AMH) are essential biomolecules produced by them during folliculogenesis.

Objective: Due to the widespread use of BPA and its potential epigenetic effects, this study examined the impact of BPA on promoter methylation of and genes in mouse granulosa cells.

Materials And Methods: Preantral follicles were isolated from ovaries of immature mice and cultured for eight days. Then, follicles were treated with 50 and 100 μM of BPA, and 0.01% (v/v) ethanol for 24 and 72 hr. Growth and degeneration of follicles and antrum formation were analyzed. The granulosa cells were isolated mechanically, and their extracted DNA was treated with sodium bisulfite. The promoter regions of the and were analyzed with PCR and sequencing.

Results: BPA did not change follicle survival and antrum formation significantly (p = 0.41). However, the culture in the presence of 100 μM BPA had an inhibitory effect on growth. Before BPA treatment, the CpG of the and promoters were unmethylated and partially methylated, respectively. While the percent of 5mC in the promoter reduced at 100 μM of BPA, it did not alter the promoter methylation.

Conclusion: BPA at higher concentrations has an inhibitory effect on follicle growth. Moreover, it seems that the epigenetic impact of BPA restricts to the demethylation of CpG sites.
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http://dx.doi.org/10.18502/ijrm.v19i2.8471DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922291PMC
February 2021

Design and characterization of new antimicrobial peptides derived from aurein 1.2 with enhanced antibacterial activity.

Biochimie 2021 Feb 30;181:42-51. Epub 2020 Nov 30.

School of Biology, Damghan University, Damghan, Iran; Institute of Biological Sciences, Damghan University, Damghan, Iran.

Antimicrobial peptides (AMPs) are promising alternative agents for treating multidrug-resistant bacterial infections. Aurein 1.2 is a natural 13-amino acid AMP with antibacterial activity against Gram-positive bacteria. In this study, we designed three novel AMPs: aurein M1 (A10W), aurein M2 (D4K, E11K), and aurein M3 (A10W, D4K, E11K) to analyze the effect of Trp substitution and enhancement of positive charge on the activity of aurein 1.2. The AMP probability, physicochemical properties, secondary and tertiary structures, and amphipathic structure were predicted by various bioinformatics tools. After the synthesis of the peptides, their antibacterial activity, hemolysis, cytotoxicity, and structural analysis were assayed. Compared to the selectivity of aurein 1.2, the selectivity of aurein M2 and M3 with a net positive charge of +5 was improved 11.30- and 8.00-fold against Gram-positive and -negative bacteria, respectively. The hemolytic activity of aurein M2 was lower than that of aurein 1.2 and M3, while the higher percentage of human fibroblast cells were alive in the presence of aurein M3. Also, the MICs of aurein M3 toward Staphylococcus aureus and Escherichia coli at the physiologic salt were ≤16, which is recommended as a promising candidate for clinical investigation. Circular dichroism analysis indicated an alpha-helical structure in the peptide analogs that is similar to aurein 1.2 in the presence of 10 mM SDS. Therefore, increasing positive charge can be used successfully as an approach for improving the potency and selectivity of AMPs. Moreover, the beneficial effect of Trp substitution depends on its position and the sequence of peptides.
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http://dx.doi.org/10.1016/j.biochi.2020.11.020DOI Listing
February 2021

Assessment of DNA Repair Gene Expressions in Vitrified Mouse Preantral Follicles.

Cell J 2020 Jul 18;22(Suppl 1):81-88. Epub 2020 Jul 18.

School of Biology, Damghan University, Damghan, Iran.

Objective: Vitrification of the ovarian tissue is one of the techniques recommended for preserving the fertility of women who are dealing with infertility. Despite its benefits, our information about the molecular aspects of ovarian follicles vitrification is somehow ambiguous. Therefore, the aim of this study was to evaluate the expression pattern of DNA repair genes in vitrified preantral follicles.

Materials And Methods: In this experimental study, the isolated preantral follicles (n=906) from 14-16 days old mice (n=12) were divided into three groups: fresh, toxic and vitrified which were cultured for 12 days. Preantral follicles were vitrified using cryotop followed by exposure to equilibration solution for five minutes and vitrification solution (VS) for 30 seconds. In the toxic group, preantral follicles were only placed in equilibration and vitrification media and they were then placed in the warming solutions without exposure to liquid nitrogen. On the second and sixth days of the culture period, real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was carried out to evaluate expression of the selected genes involved in DNA repair, including (MutS homolog 6), (Meiotic recombination 11), (Breast cancer type 1), (RAD51 recombinase), Pcna (Proliferating cell nuclear antigen) and (ATM serine/threonine kinase). In addition, developmental parameters including growth, survival rate, antrum cavity formation and ovulation were analyzed.

Results: The relative mRNA expression of and on the second and sixth days of the culture period in vitrified group was significantly higher than those of the control and toxic groups, but there was no significant difference between the toxic and control groups. In addition, developmental parameters of follicles were similar in both toxic and control groups, while both were significantly higher than that of vitrified group.

Conclusion: Vitrification changes the expression pattern of DNA repair genes of the mouse preantral follicles.
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http://dx.doi.org/10.22074/cellj.2020.6865DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7481908PMC
July 2020

Cyanobacteria as an eco-friendly resource for biofuel production: A critical review.

Biotechnol Prog 2019 09 22;35(5):e2835. Epub 2019 May 22.

Branch-Marine Science and Technology Faculty, Islamic Azad University North Tehran, Tehran, Iran.

Cyanobacteria are photosynthetic microorganisms which can be found in various environmental habitats. These photosynthetic bacteria are considered as promising feedstock for the production of the third- and the fourth-generation biofuels. The main subject of this review is highlighting the significant aspects of the biofuel production from cyanobacteria. The most recent investigations about the extraction or separation of the bio-oil from cyanobacteria are also adduced in the present review. Moreover, the genetic engineering of cyanobacteria for improving biofuel production and the impact of bioinformatics studies on the designing better-engineered strains are mentioned. The large-scale biofuel production is challenging, so the economic considerations to provide inexpensive biofuels are also cited. It seems that the future of biofuels is strongly dependent to the following items; understanding the metabolic pathways of the cyanobacterial species, progression in the construction of the engineered cyanobacteria, and inexpensive large-scale cultivation of them.
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http://dx.doi.org/10.1002/btpr.2835DOI Listing
September 2019