Publications by authors named "Hossein Vahidi"

23 Publications

  • Page 1 of 1

Emerging Antineoplastic Biogenic Gold Nanomaterials for Breast Cancer Therapeutics: A Systematic Review.

Int J Nanomedicine 2020 19;15:3577-3595. Epub 2020 May 19.

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Breast cancer remains as a concerning global health issue, being the second leading cause of cancer deaths among women in the United States (US) in 2019. Therefore, there is an urgent and substantial need to explore novel strategies to combat breast cancer. A potential solution may come from the use of cancer nanotechnology, an innovative field of study which investigates the potential of nanomaterials for cancer diagnosis, therapy, and theranostic applications. Consequently, the theranostic functionality of cancer nanotechnology has been gaining much attention between scientists during the past few years and is growing exponentially. The use of biosynthesized gold nanoparticles (AuNPs) has been explored as an efficient mechanism for the treatment of breast cancer. The present study supposed a global systematic review to evaluate the effectiveness of biogenic AuNPs for the treatment of breast cancer and their anticancer molecular mechanisms through in vitro studies. Online electronic databases, including Cochrane, PubMed, Scopus, Web of Science, Science Direct, ProQuest, and Embase, were searched for the articles published up to July 16, 2019. Our findings revealed that plant-mediated synthesis was the most common approach for the generation of AuNPs. Most of the studies reported spherical or nearly spherical-shaped AuNPs with a mean diameter less than 100 nm in size. A significantly larger cytotoxicity was observed when the biogenic AuNPs were tested towards breast cancer cells compared to healthy cells. Moreover, biogenic AuNPs demonstrated significant synergistic activity in combination with other anticancer drugs through in vitro studies. Although we provided strong and comprehensive preliminary in vitro data, further in vivo investigations are required to show the reliability and efficacy of these NPs in animal models.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2147/IJN.S240293DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7245458PMC
August 2020

Fungus-mediated Extracellular Biosynthesis and Characterization of Zirconium Nanoparticles Using Standard Species and Their Preliminary Bactericidal Potential: A Novel Biological Approach to Nanoparticle Synthesis.

Iran J Pharm Res 2019 ;18(4):2101-2110

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Biological synthesis of nanoparticles (NPs) has gained extensive attention during recent years by using various biological resources such as plant extracts and microorganisms as reducing and stabilizing agents. The objective of the present study was to biosynthesize zirconium NPs using species as a reliable and eco-friendly protocol for the first time. The synthesized NPs were characterized using Scanning Electron Microscope (SEM), Atomic Force Microscope (AFM), Dynamic Light Scattering (DLS), Energy Dispersive X-ray (EDX), and Fourier Transform Infrared (FT-IR). The results showed that three species were able to synthesize zirconium NPs extracellularly with spherical morphology below 100 nm. Moreover, the preliminary antibacterial activity of zirconium NPs represented considerable antibacterial potential against Gram-negative bacteria. Overall, the current study demonstrated a novel bio-based approach for preparation of zirconium NPs. Further studies are required to expend this laboratory-based investigation to an industrial scale owing to their superiorities over traditional physicochemical methods such as cost-effectiveness and eco-friendliness.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.22037/ijpr.2019.112382.13722DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7059062PMC
January 2019

A Systematic Review of the Genotoxicity and Antigenotoxicity of Biologically Synthesized Metallic Nanomaterials: Are Green Nanoparticles Safe Enough for Clinical Marketing?

Medicina (Kaunas) 2019 Aug 5;55(8). Epub 2019 Aug 5.

Pharmaceutical Sciences Research Center, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari 48175 861 Iran.

Although studies have elucidated the significant biomedical potential of biogenic metallic nanoparticles (MNPs), it is very important to explore the hazards associated with the use of biogenic MNPs. Evidence indicates that genetic toxicity causes mutation, carcinogenesis, and cell death. Therefore, we systematically review original studies that investigated the genotoxic effect of biologically synthesized MNPs via in vitro and in vivo models. Articles were systematically collected by screening the literature published online in the following databases; Cochrane, Web of Science, PubMed, Scopus, Science Direct, ProQuest, and EBSCO. : Most of the studies were carried out on the MCF-7 cancer cell line and phytosynthesis was the general approach to MNP preparation in all studies. Fungi were the second most predominant resource applied for MNP synthesis. A total of 80.57% of the studies synthesized biogenic MNPs with sizes below 50 nm. The genotoxicity of Ag, Au, ZnO, TiO, Se, Cu, Pt, Zn, Ag-Au, CdS, FeO, TbO, and Si-Ag NPs was evaluated. AgNPs, prepared in 68.79% of studies, and AuNPs, prepared in 12.76%, were the two most predominant biogenic MNPs synthesized and evaluated in the included articles. : Although several studies reported the antigenotoxic influence of biogenic MNPs, most of them reported biogenic MNP genotoxicity at specific concentrations and with a dose or time dependence. To the best of our knowledge, this is the first study to systematically evaluate the genotoxicity of biologically synthesized MNPs and provide a valuable summary of genotoxicity data. In conclusion, our study implied that the genotoxicity of biologically synthesized MNPs varies case-by-case and highly dependent on the synthesis parameters, biological source, applied assay, etc. The gathered data are required for the translation of these nanoproducts from research laboratories to the clinical market.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/medicina55080439DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722661PMC
August 2019

Optimization of Growth Conditions of Mycelium and Polysaccharides on Walnut Shell by-Products Using Response Surface Analysis.

Iran J Pharm Res 2018 ;17(4):1509-1522

Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

() is one of the most widely used traditional Chinese medicines and a high producer of various bioactive compounds such as polysaccharides. It has been shown that polysaccharides (LEPLS) have several physiological effects with potential medical applications. In addition, the ability of to grow and produce bioactive compounds on industrial by-products makes it an excellent candidate for the lage-scale production of such compounds. The objective of this study was to optimize mycelium and polysaccharide production by on walnut shell through a two-step procedure including a one-factor-at-a-time approach to select the most important factors and a response surface methodology design to determine their optimum combinations. Several factors were evaluated in the first step and among them inoculum size, incubation time, and C/N ratio were selected for optimization of using RSM. The RSM model estimated that a maximal yield of biomass and LEPLS (0.043 mg/g and 46.80 mg/g respectively) could be obtained when inoculum size, incubation time, and C/N ratio were set at 23.41, 30, 10 units, respectively. These values were also verified by validation experiments.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6269588PMC
January 2018

An efficient biotransformation of progesterone into 11α-hydroxyprogesterone by Rhizopus microsporus var. oligosporus.

Z Naturforsch C J Biosci 2018 Dec;74(1-2):9-15

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, P.O. Box 14155-6153, Tehran, Iran.

Rhizopus microsporus var. oligosporus is a fungus that belongs to the Mucoraceae family that is used for the preparation of some soy-fermented foods. Microbial biotransformation of progesterone by R. microsporus var. oligosporus afforded some monohydroxylated and dihydroxylated metabolites. The main product was purified using chromatographic methods and identified as 11α-hydroxyprogesterone on the basis of its spectroscopic features. Time course studies by high-performance thin-layer chromatography demonstrated that this fungi efficiently hydroxylated progesterone at the 11α-position for 3 days with a yield of 76.48%, but beyond this time, the microorganism transformed 11α-hydroxyprogesterone into dihydroxylated metabolites. 11α-Hydroxyprogesterone is widely used as a precursor in the synthesis of hydrocortisone and other steroidal anti-inflammatory agents.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1515/znc-2018-0092DOI Listing
December 2018

Effect of Acyl Homoserine Lactone on Recombinant Production of Human Insulin-like Growth Factor-1 in Batch Culture of Escherichia coli.

Protein Pept Lett 2018 ;25(11):980-985

Protein Technology Research Center, Shahid Behashti University of Medical Sciences, Tehran, Iran.

Background: IGF-I as a human growth factor produced in Escherichia coli is a single, non-glycosylated, polypeptide chain containing 70 amino acids and having a molecular mass of 7.6 kDa. Up to now, E. coli expression system has been widely used as the host to produce rhIGF-1 with high yields. Acyl Homoserine Lactones (AHLs) are intercellular signaling molecules used in quorum sensing by Gram-negative bacteria. Quorum sensing is a cell density-dependent gene regulation process that allows bacterial cells to express specific genes only when signaling molecules reach the sufficient concentration.

Objective: For the first time, this study focuses on the N-hexanoyl-L- Homoserine Lactone (HHL) activity on increasing the cell growth and rh-IGF-1concentration in batch culture of E. coli.

Method: The maximum production of rhIGF-I was previously optimized in 32y culture medium at 32°C with 0.05 mM IPTG as inducer and 10 g/l glucose concentration. Under this condition, different amounts of HHL (0.001 µg/ml, 1 µg/ml, and 100µg/ml) were evaluated as an inducer for IGF-1 production.

Results: Generally, with increasing of HHL concentration, an increase in dry cell weight (2.45 mg/ml to 4.63 mg/ml) and IGF-I expression level (0.4 mg/ml to 0.77 mg/ml) was observed.

Conclusion: HHL or other types of AHLs can be considered as protein production inducer in bacterial expression systems through the quorum sensing pathways.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2174/0929866525666181019150657DOI Listing
January 2019

Simultaneous Optimization of the Production of Organic Selenium and Cell Biomass in by Plackett-Burman and Box-Behnken Design.

Iran J Pharm Res 2018 ;17(3):1081-1092

Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Selenium (Se) as a vital trace element has many biological activities such as anti-inflammation and anti-oxidation. Selenomethionine as an organic selenium plays a vital role in the response to oxidative stress. At present, is one of the best microorganisms that has the ability to accumulate selenium. Production of Seleno-yeast was done by growing in the presence of water soluble selenium salt (NaSeO) as a part of the medium. The yield of selenium biotransformation and yeast biomass can be improved by optimizing the process conditions in two steps. First, the effects of several culture parameters (culture conditions and culture media) were studied using the Plackett-Burman design. After that, determining the optimum levels of the effective parameters was performed by Box-Behnken response surface methodology. Optimization of the conditions was performed with the aim of simultaneously optimizing the biomass and selenium biotransformation. In this investigation, the effect of the eleven culture parameters was studied with Plackett-Burman design. Then, four significant culture parameters such as glucose concentration, aeration, selenium concentration, and temperature were optimized with Box-Behnken response surface methodology.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6094416PMC
January 2018

Baeyer-Villiger oxidation of progesterone by Aspergillus sojae PTCC 5196.

Steroids 2018 12 26;140:52-57. Epub 2018 Jul 26.

Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran 1417614411, Iran.

Microbial transformations are capable of producing steroid substances difficult to synthesize by chemical methods. Strains belonging to the genus Aspergillus are effective facilitators of microbial biotransformations due to their enzymatic diversity. In this study, the biotransformation of progesterone by the fungus Aspergillus sojae (A. sojae) PTCC 5196 was examined. Analysis of the bioconversion process revealed that progesterone was converted to testololactone through a three-step pathway (17β-acetyl side chain cleavage, 17β-hydroxyl oxidation, and oxygenative lactonization of 17-ketone), indicating the presence of Baeyer-Villiger monooxygenase (BVMO) activity in the fungal strain. GC analysis confirmed the production of testololactone with a yield of 99% in 24 h. Faster testololactone production was induced in the presence of both C-21 (progesterone) and C-19 (androstenedione, testosterone, and dehydroepiandrosterone [DHEA]) steroid substances. Due to the high biotransformation rate observed in the present study, A. sojae may be a novel and promising candidate in the production of testololactone.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.steroids.2018.07.008DOI Listing
December 2018

Biosynthesis and Characterization of Biogenic Tellurium Nanoparticles by Using PTCC 5031: A Novel Approach in Gold Biotechnology.

Iran J Pharm Res 2018 ;17(Suppl2):87-97

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Production of nanoparticles has been attractive by biological based fabrication as an alternative to physical and chemical approaches due to exceeding need to develop safe, reliable, clean and eco-friendly methods for the preparation of nanoparticle for pharmaceutical and biomedical applications. In the present study, biogenic tellurium nanoparticles (TeNPs) were successfully prepared using potassium tellurite (KTeO, 3HO) via an eco-friendly and simple green approach by exploiting extracellular enzymes and biomolecules secreted from PTCC 5031 at room temperature for the first time. The biofabricated TeNPs were characterized by Atomic Force Microscope (AFM), Scanning Electron Microscopy (SEM), Dynamic Light Scattering (DLS), Energy Dispersive X-ray spectroscopy (EDX), and Fourier Transform Infrared (FT-IR) spectrum. The AFM and SEM images revealed that the TeNPs were fairly uniform in size with a spherical shape and superior monodispersity. Furthermore, the DLS indicated that the average hydrodynamic diameter of TeNPs was around 50.16 nm and polydispersity index (PdI) of 0.012. The EDX results depicted that TeNPs display an absorption peak at 3.8 keV, indicating the presence of the elemental tellurium. Additionally, the FT-IR analysis of TeNPs exhibited the presence of possible functional groups that may have a role as bioreducers and capping agents. Overall, the results strongly suggested that can be a potential nanofactory for the preparation of TeNPs due to several advantages including non-pathogenic organism, fast growth rate, and high capacity of elemental ions reduction, as well as facile and economical biomass handling.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447880PMC
January 2018

Challenges to Design and Develop of DNA Aptamers for Protein Targets. II. Development of the Aptameric Affinity Ligands Specific to Human Plasma Coagulation Factor VIII Using SEC-SELEX.

Iran J Pharm Res 2017 ;16(2):737-744

Protein Technology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Protein specific aptamers are highly applicable affinity ligands in different fields of research and clinical applications. They have been developed against various targets, in particular, bio-macromolecules such as proteins. Among human proteins, the coagulation factors are the most attractive targets for aptamer selection and their specific aptamers have valuable characteristics in therapeutic and analytical applications. In this study, a plasma derived coagulation factor VIII was considered as the protein target for DNA aptamer selection using size exclusion chromatography-SELEX. Potential aptameric oligonucleotides with high affinity and specificity were achieved during eight rounds of selection. Binding affinity constant of selected aptamer and aptameric enriched pool were in nanomolar range that was comparable to monoclonal antibodies. Further improvement studies can result in aptamers that are more promising as an industrial affinity ligand for the purification of anti-hemophilia factor from plasma source.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5603883PMC
January 2017

RP-HPLC Method Development and Validation for Determination of Eptifibatide Acetate in Bulk Drug Substance and Pharmaceutical Dosage Forms.

Iran J Pharm Res 2017 ;16(2):490-497

Phytochemistry Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

A new, rapid, economical and isocratic reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of eptifibatide acetate, a small synthetic antiplatelet peptide, in bulk drug substance and pharmaceutical dosage forms. The developed method was validated as per of ICH guidelines. The chromatographic separation was achieved isocratically on C18 column (150 x 4.60 mm i.d., 5 µM particle size) at ambient temperature using acetonitrile (ACN), water and trifluoroacetic acid (TFA) as mobile phase at flow rate of 1 mL/min and UV detection at 275 nm. Eptifibatide acetate exhibited linearity over the concentration range of 0.15-2 mg/mL (r=0.997) with limit of detection of 0.15 mg/mL The accuracy of the method was 96.4-103.8%. The intra-day and inter-day precision were between 0.052% and 0.598%, respectively. The present successfully validated method with excellent selectivity, linearity, sensitivity, precision and accuracy was applicable for the assay of eptifibatide acetate in bulk drug substance and pharmaceutical dosage forms.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5603858PMC
January 2017

L-Asparaginase Activity in Cell Lysates and Culture Media of Halophilic Bacterial Isolates.

Iran J Pharm Res 2016 ;15(3):435-440

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

The objective of this study was to isolate halophilic bacteria with the ability to produce intracellular or extracellular L-asparaginase. A total number of 120 halophilic bacteria were isolated from 17 different saline habitats of Iran including salt lakes, wetlands, brine springs and deserts. Among these, 68 were able to grow in the presence of 1.5 M NaCl and 52 demonstrated the ability to grow in the selection medium containing 3.5 M NaCl. None of the isolates appeared to produce appreciable amounts of extracellular L-asparaginase. Among the isolates that produced intracellular L-asparaginase, 5 moderate and 1 extreme halophiles were selected for further study based on their observed activity level. The moderately halophilic isolates were shown to belong to the genus while the extreme halophile was identified as a member of the genus .
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5149030PMC
January 2016

Biotransformation of Progesterone by Whole Cells of Filamentous Fungi Aspergillus brasiliensis.

Iran J Pharm Res 2015 ;14(3):919-24

Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ; Phytochemistry Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Microbial steroid biotransformations have found a wide-reaching application for the production of more precious and functionalized compounds due to their high regio-and stereo selectivity. In this study, the possibility of using filamentous fungi Aspergillus brasiliensiscells in the biotransformation of progesterone, a C-21 steroid hormone was studied for the first time. The fungal strain was inoculated into the transformation medium supplemented with progesterone as a substrate. Biotransformation of this steroid for 7 days afforded 3 different hydroxylated metabolites: 11α-hydroxy progesterone; 14α-hydroxyprogesteroneand21-hydroxyprogesterone. The metabolites were separated by thin layer chromatography. Structure determinations of the metabolites were performed by comparing NMR, MS and IR spectra of the starting compound with those of metabolites. These results may be of industrial importance because the metabolites can be used as precursor of some steroid drugs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4518121PMC
September 2015

Enhanced Production of Insulin-like Growth Factor I Protein in Escherichia coli by Optimization of Five Key Factors.

Iran J Pharm Res 2015 ;14(3):907-17

Department of Life Science Engineering, School of New Science and Technologies, University of Tehran, Iran.

Human insulin-like growth factor I (hIGF-I) is a kind of growth factor with clinical significance in medicine. Up to now, E. coli expression system has been widely used as a host to produce rhIGF-1 with high yields. Batch cultures as non-continuous fermentations were carried out to overproduce rhIGF-I in E. coli. The major objective of this study is over- production of recombinant human insulin-like growth factor I (rhIGF-I) through a developed process by recruiting effective factors in order to achieve the most recombinant protein. In this study we investigated the effect of culture medium, induction temperature and amount of inducer on cell growth and IGF-1 production. Taguchi design of experiments (DOE) method was used as the statistical method. Analysis of experimental data showed that maximum production of rhIGF-I was occurred in 32y culture medium at 32 °C and 0.05 Mm IPTG. Under this condition, 0.694 g/L of rhIGF-I was produced as the inclusion bodies. Following optimization of these three factors, we have also optimized the amount of glucose and induction time in 5 liter top bench bioreactor. Full factorial design of experiment method was used for these two factors as the statistical method. 10 g/L and OD600=5 were selected as the optimum point of Glucose amount and induction time, respectively. Finally, we reached to a concentration of 1.26 g/L rhIGF-1 at optimum condition.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4518120PMC
September 2015

Optimization of Fermentation Conditions for Recombinant Human Interferon Beta Production by Escherichia coli Using the Response Surface Methodology.

Jundishapur J Microbiol 2015 Apr 18;8(4):e16236. Epub 2015 Apr 18.

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran.

Background: The periplasmic overexpression of recombinant human interferon beta (rhIFN-β)-1b using a synthetic gene in Escherichia coli BL21 (DE3) was optimized in shake flasks using Response Surface Methodology (RSM) based on the Box-Behnken Design (BBD).

Objectives: This study aimed to predict and develop the optimal fermentation conditions for periplasmic expression of rhIFN-β-1b in shake flasks whilst keeping the acetate excretion as the lowest amount and exploit the best results condition for rhIFN-β in a bench top bioreactor.

Materials And Methods: The process variables studied were the concentration of glucose as carbon source, cell density prior the induction (OD 600 nm) and induction temperature. Ultimately, a three-factor three-level BBD was employed during the optimization process. The rhIFN-β production and the acetate excretion served as the evaluated responses.

Results: The proposed optimum fermentation condition consisted of 7.81 g L(-1) glucose, OD 600 nm prior induction 1.66 and induction temperature of 30.27°C. The model prediction of 0.267 g L(-1) of rhIFN-β and 0.961 g L(-1) of acetate at the optimum conditions was verified experimentally as 0.255 g L(-1) and 0.981 g L(-1) of acetate. This agreement between the predicted and observed values confirmed the precision of the applied method to predict the optimum conditions.

Conclusions: It can be concluded that the RSM is an effective method for the optimization of recombinant protein expression using synthetic genes in E. coli.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5812/jjm.8(4)2015.16236DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4449858PMC
April 2015

Cloning and Expression of TNF Related Apoptosis Inducing Ligand in Nicotiana tabacum.

Iran J Pharm Res 2015 ;14(1):189-201

Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

Molecular farming has been considered as a secure and economical approach for production of biopharmaceuticals. Human TNF Related Apoptosis Inducing Ligand (TRAIL) as a promising biopharmaceutical candidate has been produced in different expression hosts. However, little attention has been paid to molecular farming of the TRAIL in spite of numerous advantages of plant expression systems. Therefore, in this study the cytoplasmic production of the TRAIL was tackled in Nicotiana tabacum using Agrobacterium tumefaciens LBA 4404. Initially, the desired coding sequence was obtained using PCR technique on the constructed human cDNA library. Afterward, the necessary requirements for expression of the TRAIL in plant cell system were provided through sub-cloning into 35S-CaMV (Cauliflower Mosaic Virus) helper and final 0179-pGreen expression vectors. Then, the final TRAIL-pGreen expression vector was cloned into A. tumefaciens LBA 4404. Subsequently, the N. tabacum cells were transformed through co-culture method and expression of the TRAIL was confirmed by western blot analysis. Finally, the recombinant TRAIL was extracted through chromatographic technique and biological activity was evaluated through MTT assay (Methylthiazol Tetrazolium Assay). The result of western blot analysis indicated that only monomer and oxidized dimer forms of the TRAIL can be extracted from the N. tabacum cells. Moreover, the lack of trimeric assembly of the extracted TRAIL diminished its biological activity in sensitive A549 cell line. In conclusion, although N. tabacum cells can successfully produce the TRAIL, proper assembly and functionality of the TRAIL were unfavorable.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4277632PMC
January 2015

Fungal transformation of androsta-1,4-diene-3,17-dione by Aspergillus brasiliensis.

Daru 2014 Nov 15;22:71. Epub 2014 Nov 15.

Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Vali-e Asr Ave., Niayesh Junction, Tehran, 1996835113, Iran.

Background: The biotransformation of steroids by fungal biocatalysts has been recognized for many years. There are numerous fungi of the genus Aspergillus which have been shown to transform different steroid substances. The possibility of using filamentous fungi Aspergillus brasiliensis cells in the biotransformation of androsta-1,4-diene-3,17-dione, was evaluated.

Methods: The fungal strain was inoculated into the transformation medium which supplemented with androstadienedione as a substrate and fermentation continued for 5 days. The metabolites were extracted and isolated by thin layer chromatography. The structures of these metabolites were elucidated using (1)H-NMR, broadband decoupled (13)C-NMR, EI Mass and IR spectroscopies.

Results: The fermentation yielded one reduced product: 17β-hydroxyandrost-1,4-dien-3-one and two hydroxylated metabolites: 11α-hydroxyandrost-1,4-diene-3,17-dione and 12β-hydroxyandrost-1,4-diene-3,17-dione.

Conclusions: The results obtained in this study show that A. brasiliendsis could be considered as a biocatalyst for producing important derivatives from androstadienedione.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s40199-014-0071-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4241229PMC
November 2014

Improvement in the stability and functionality of Nicotiana tabacum produced recombinant TRAIL through employment of endoplasmic reticulum expression and ascorbate buffer mediated extraction strategies.

Bioimpacts 2014 17;4(3):123-32. Epub 2014 May 17.

Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

Introduction: In order to employ Nicotiana tabacum cells as a profitable natural bioreactor for production of bio-functional "Soluble human TRAIL" (ShTRAIL), endoplasmic reticulum (ER) targeted expression and innovative extraction procedures were exploited.

Methods: At first, the ShTRAIL encoding gene was sub-cloned into designed H2 helper vector to equip it with potent TMV omega leader sequences, ER sorting signal peptide, poly-histidine tag and ER retention signal peptide (KDEL). Then, the ER targeted ShTRAIL cassette was sequentially sub-cloned into "CaMV-35S" helper and "pGreen-0179" final expression vectors. Afterward, Agrobacterium mediated transformation method was adopted to express the ShTRAIL in the ER of N. tabacum . Next, the ShTRAIL protein was extracted through both phosphate and innovative ascorbate extraction buffers. Subsequently, oligomerization state of the ShTRAIL was evaluated through cross-linking assay and western blot analysis. Then, semi-quantitative western blot analysis was performed to estimate the ShTRAIL production. Finally, biological activity of the ShTRAIL was evaluated through MTT assay.

Results: The phosphate buffer extracted ShTRAIL was produced in dimmer form, whereas the ShTRAIL extracted with ascorbate buffer generated trimer form. The ER targeted ShTRAIL strategy increased the ShTRAIL's production level up to about 20 μg/g of fresh weight of N. tabacum . MTT assay indicated that ascorbate buffer extracted ShTRAIL could prohibit proliferation of A549 cell line.

Conclusion: Endoplasmic reticulum expression and reductive ascorbate buffer extraction procedure can be employed to enhance the stability and overall production level of bio-functional recombinant ShTRAIL from transgenic N. tabacum cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.15171/bi.2014.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4204037PMC
October 2014

Overexpression of Recombinant Human Beta Interferon (rhINF-β) in Periplasmic Space of Escherichia coli.

Iran J Pharm Res 2014 ;13(Suppl):151-60

Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran.

Human Interferon β (INF-β) is a member of cytokines family which different studies have shown its immunomodulatory and antiviral activities. In this study an expression vector was designed and constructed for expression of human INF-β-1b either in shake flasks or bench top bioreactor. The designed vector was constructed based upon pET-25b(+) with T7 promoter. Recombinant human beta interferon (rhINF-β) was codon optimized and overexpressed as a soluble, N-terminal pelB fusion protein and secreted into the periplasmic space of Escherichia coli BL21 (DE3). The sugar, Isopropyl-β-D-thiogalactopyranoside (IPTG) was used as a chemical inducer for rhINF-β production in the shake flasks and bench top bioreactor. Timing of beta interferon expression was controlled by using the T7 promoter. The rhINF-β protein was extracted from periplasmic space by osmotic shock treatment and the expression of the beta interferon encoding gene in random selected transformants, was confirmed by western and dot blot methods. The maximum of product formation achieved at the OD600nm = 3.42 was found to be 35 % of the total protein content of the strain which translates to 0.32 g L-1. The constructed vector could efficiently overexpress the rhINF-β into the periplasmic space of E. coli. The obtained yield of the produced rhINF-β was more than previous reports. The system is easily adapted to include other vectors, tags or fusions and therefore has the potential to be broadly applicable to express other recombinant proteins.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3977065PMC
April 2014

Challenges to design and develop of DNA aptamers for protein targets. I. Optimization of asymmetric PCR for generation of a single stranded DNA library.

Iran J Pharm Res 2014 ;13(Suppl):133-41

Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. Asymmetric PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of aptamers with high affinity for protein targets and develop an efficient asymmetric PCR amplification. Thus, the influence of factors including annealing temperature, number of amplification cycles, primer ratio, Mg(2+) concentration and the presence of a PCR enhancer on the amplification of the desired product were evaluated. Results obtained by agarose gel electrophoresis showed that the annealing temperature of 64 °C, Mg(2+) concentration of 0.25 mM, reverse to forward primer ratio of 15:1, amplification cycle of 25 and the presence L-ectoin as a PCR enhancer with the concentration of 0.4 M were the optimal conditions. Our results supported that the yield of this type of ssDNA production is sufficient for combinatorial screening of aptamers.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3977063PMC
April 2014

25 Years of On-going Endeavors and Experience towards Promoting Iran's Pharmacy.

Authors:
Hossein Vahidi

Iran J Pharm Res 2013 ;12(Suppl)

View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3813378PMC
November 2013

Detection of genetically modified maize in processed foods sold commercially in iran by qualitative PCR.

Iran J Pharm Res 2013 ;12(1):25-30

Food and Drug Laboratory Research Center, Food and Drug Organization, Ministry of Health and Medical Education, Tehran, Iran. ; Department of Molecular Biology, Food and Drug Control Reference Laboratory, Ministry of Health and Medical Education, Tehran, Iran.

Detection of genetically modified organisms (GMOs) in food is an important issue for all the subjects involved in food control and customer's right. Due to the increasing number of GMOs imported to Iran during the past few years, it has become necessary to screen the products in order to determine the identity of the consumed daily foodstuffs. In this study, following the extraction of genomic DNA from processed foods sold commercially in Iran, qualitative PCR was performed to detect genetically modified maize. The recombinant DNA target sequences were detected with primers highly specific for each investigated transgene such as CaMV35s gene, Bt-11, MON810 and Bt-176 separately. Based on the gel electrophoresis results, Bt- 11 and MON810 events were detected in some maize samples, while, in none of them Bt- 176 modified gene was detected. For the first time, the results demonstrate the presence of genetically modified maize in Iranian food products, reinforcing the need for the development of labeling system and valid quantitative methods in routine analyses.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3813200PMC
November 2013

Degradation of petroleum aromatic hydrocarbons using TiO2 nanopowder film.

Environ Technol 2013 May-Jun;34(9-12):1183-90

Department of Civil & Environmental Engineering, Graduate Faculty of Environment, University of Tehran, Tehran, Iran.

The performance of a photo-reactor packed with titanium dioxide (TiO2) immobilized on glass beads, initiated by irradiation with natural and artificial ultraviolet (UV) sources, was evaluated in terms of the degradation efficiency of petroleum aromatic hydrocarbons. The effects of parameters such as pH, reaction time, hydrogen peroxide (H2O2) concentration and some ions were investigated. Additionally, the degradation of total organic carbon (TOC) and the formation of byproducts were studied. Photodegradation rates ofbenzene, toluene, ethylbenzene and xylenes (BTEX) by processes of UV/TiO2 and UV/TiO2/H2O2 were found to obey pseudo first-order kinetic models. Results indicated that the effect of pH value was negligible at the pH range of 5.5 to 8.5. TOC removal improved with addition of H2O2 demonstrating that a lack of hydrogen peroxide leads to incomplete mineralization. The effect of cations and anions on the photodegradation efficiencies of BTEX revealed that Mg2+ and Ca2+ caused the most deterioration in BTEX degradation efficiency. However S4O(2-) and CO3(2-) had the most salient inhibitory effects compared with other tested anions. The degradation efficiencies of both systems were investigated for the treatment of real polluted groundwater collected from the city of Tehran. Results showed that the degradation efficiencies of BTEX declined in the presence of inorganic and organic competitor species.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/09593330.2012.743592DOI Listing
December 2013