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Habibi Z.,Shahid Beheshti University | Mohammadi M.,Iran National Institute of Genetic Engineering and Biotechnology | Yousefi M.,Avicenna Research Institute
Process Biochemistry | Year: 2013

This research describes the immobilization of Rhizomucor miehei lipase (RML) and chemically aminated RML (NH2-RML) on different supports including octyl-sepharose (octyl-RML), activated sepharose with cyanogen bromide (CNBr-RML and CNBr-NH2-RML), glyoxyl sepharose (Gx-RML and Gx-NH2-RML) and glyoxyl sepharose dithiothreitol (Gx-DTT-RML and Gx-DTT-NH2-RML). The highest immobilization yield was achieved for octyl-RML (>98%) followed by CNBr-RML (88%). Octyl-RML had the most specific activity (13.6) among all derivatives. The other preparations had moderate activities likely because of chemical reaction during covalent attachment of the enzyme. The catalytic behavior of lipase immobilized in hydrolysis reactions was investigated using methyl, ethyl, propyl, butyl and isobutyl-ibuprofen esters and the influence of the alkyl chain and the alcoholic residue of the ester were studied. Butyl ester was the most interesting ester for carrying out hydrolysis. The highest enantioselectivity of enzyme (E = 8.8) was obtained with isooctane/sodium phosphate buffer pH 7.0 at temperature of 40 °C. Increasing temperature from 40 to 50 °C caused decreasing in enantioselectivities and conversions. Also esterification of ibuprofen was carried out in solvent systems containing isooctane and two ionic liquids (ILs); [BMIM][PF6] and [BMIM][BF4]. Poor conversions and enantioselectivities were observed during esterification in all solvents. © 2013 Elsevier Ltd.


Kalantari M.,Iran National Institute of Genetic Engineering and Biotechnology | Kalantari M.,Sharif University of Technology | Kazemeini M.,Sharif University of Technology | Tabandeh F.,Iran National Institute of Genetic Engineering and Biotechnology | Arpanaei A.,Iran National Institute of Genetic Engineering and Biotechnology
Journal of Materials Chemistry | Year: 2012

Uniformly sized superparamagnetic single-shell nonporous (S1) and double-shell mesoporous silica nanocomposite particles with ∼130 nm magnetite cluster cores are synthesised in this study. Mesoporous particles are prepared with two BJH pore sizes (2.44 and 3.76 nm, designated as S2 and S3 particles, respectively). Once the lipase was immobilised on particles, our results showed that the enzyme loading capacities of mesoporous structures, i.e. S2 and S3, are higher than that for nonporous particles (S1). Hydrolytic activity tests reveal that immobilised lipases retain about 90% of the free enzyme's activity. Furthermore, comparing to the free enzyme, the thermal stability of immobilised enzymes is considerably enhanced, regardless of the silica matrix structure. Also, our results indicate that the rate of the enzymatic reaction is not influenced by the type of silica matrix used for lipase immobilisation, but immobilisation results in lower enzymatic reaction rates for the immobilised enzymes compared to the free enzyme. Finally, in examining the reusability, the immobilised lipases retained more than 76% of their initial activities after 5 times' reuse. © The Royal Society of Chemistry 2012.


Mahmoudi M.,Pasteur Institute of Iran | Mahmoudi M.,Tehran University of Medical Sciences | Laurent S.,University of Mons | Shokrgozar M.A.,Pasteur Institute of Iran | And 2 more authors.
ACS Nano | Year: 2011

In the last few decades, nanoparticles (NPs) have been recognized as promising candidates for starting a new revolution in science and technology due to their unusual properties, attracting the attention of physicists, chemists, biologists, and engineers. The aim of this study is to evaluate the toxicities (at both cellular and molecular levels) of three forms of superparamagnetic iron oxide nanoparticles (SPIONs) of various surface chemistries (COOH, plain, and NH2) through the comparison with gene expression patterns of three cell types (i.e., human heart, brain, and kidney). For this purpose, both an MTT assay and a DNA microarray analysis were applied in three human cell lines-HCM (heart), BE-2-C (brain), and 293T (kidney)-under the exposure to SPIONs-COOH, SPIONs-NH2, and bare SPIONs. The specific gene alteration and hierarchical clustering revealed that SPIONs-COOH altered genes associated with cell proliferative responses due to their reactive oxygen species (ROS) properties. It was also found that the cell type can have quite a significant role in the definition of suitable pathways for detoxification of NPs, which has deep implications for the safe and high yield design of NPs for biomedical applications and will require serious consideration in the future. © 2011 American Chemical Society.


Tabasi O.,Amirkabir University of Technology | Falamaki C.,Amirkabir University of Technology | Khalaj Z.,Iran National Institute of Genetic Engineering and Biotechnology
Colloids and Surfaces B: Biointerfaces | Year: 2012

The present work concerns a preliminary step in the production of anticancer drug loaded porous silicon (PSi) for targeted-drug-delivery applications. A successful procedure for the covalent attachment of folic acid, polyethylene glycol (PEG) and doxorubicin to hydrophilic mesoporous silicon layers is presented. A systematic approach has been followed to obtain the optimal composition of the N,N'-dicyclohexylcarbodiimide (DCC)/N-hydroxysuccimide (NHS) in dimethylsulfoxide (DMSO) solution for the surface activation process of the undecylenic acid (UD) grafted molecules to take place with minimal undesired byproduct formation. The effect of reactant concentration and kind of solvent (aqueous or DMSO) on the attachment of folic acid to the activated PSi layer has been investigated. The covalent attachment of the doxorubicin molecules to the PSi layer functionalized with folic acid and PEG is discussed. The drug release kinetics as a function of pH has been studied. The functionalized PSi particles show a high cytotoxicity compared to the equivalent amount of free drug. Cell toxicity tests show clearly that the incorporation of folate molecules increases substantially the toxicity of the loaded PSi particles. Accordingly this new functionalized PSi may be considered a proper candidate for targeted drug delivery. © 2012 Elsevier B.V.


Taha M.F.,Iran National Institute of Genetic Engineering and Biotechnology
Current Stem Cell Research and Therapy | Year: 2010

Cell based-gene delivery has provided an important therapeutic strategy for different disorders in the recent years. This strategy is based on the transplantation of genetically modified cells to express specific genes and to target the delivery of therapeutic factors, especially for the treatment of cancers and neurological, immunological, cardiovascular and heamatopoietic disorders. Although, preliminary reports are encouraging, and experimental studies indicate functionally and structurally improvements in the animal models of different disorders, universal application of this strategy for human diseases requires more evidence. There are a number of parameters that need to be evaluated, including the optimal cell source, the most effective gene/genes to be delivered, the optimal vector and method of gene delivery into the cells and the most efficient route for the delivery of genetically modified cells into the patient. Also, some obstacles have to be overcome, including the safety and usefulness of the approaches and the stability of the improvements. Here, recent studies concerning with the cell-based gene delivery for spinal cord injury and some neurodegenerative disorders such as amyotrophic lateral sclerosis, Parkinson's disease and Alzheimer's disease are briefly reviewed, and their exciting consequences are discussed. © 2010 Bentham Science Publishers Ltd.


Gashtasbi F.,Iran National Institute of Genetic Engineering and Biotechnology | Ahmadian G.,Iran National Institute of Genetic Engineering and Biotechnology | Noghabi K.A.,Iran National Institute of Genetic Engineering and Biotechnology
Enzyme and Microbial Technology | Year: 2014

Most of the studies in the field of enzyme immobilization have sought to develop a simple, efficient and cost-effective immobilization system. In this study, probiotic Bacillus spores were used as a matrix for enzyme immobilization, because of their inherent resistance to extreme temperatures, UV irradiation, solvents and drying. Above all, their preparation is cost-effective. The alpha-amylase enzyme was immobilized on the spore surface by the covalent and adsorption methods. For the covalent method, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N hydroxysulfosuccinimide (NHS) were used. The maximum concentration of the alpha-amylase immobilized by the two methods onto the spore surface was 360μg/1.2×1011 spore. However, maximum activity was achieved at an enzyme concentration of approximately 60μg/.4×1010, corresponding to an estimated activity of 8×103IUmg-1/1.2×1011 spore for covalent immobilization and 8.53×103 for the adsorption method. After washing the enzyme with 1M NaCl and 0.5% Triton X-100, the enzyme immobilization yield was estimated to be 77% and 20.07% for the covalent and adsorption methods, respectively. The alpha-amylase immobilized by both methods, displayed improved activity in the basic pH range. The optimum pH for the free enzyme was 5 while it shifted to 8 for the immobilized enzyme. The optimum temperatures for the free and immobilized enzymes were 60°C and 80°C, respectively. The covalently-immobilized alpha-amylase retained 65% of its initial activity, even after 10 times of recycling. The Km and Vmax values were determined by the GraphPad Prism software, which showed that the Vmax value decreased moderately after immobilization. This is the first study which reports the covalent immobilization of an enzyme on the spore surface. © 2014 Elsevier Inc.


Akhavan O.,Sharif University of Technology | Ghaderi E.,Nanobiotechnology Research Laboratory | Hashemi E.,Iran National Institute of Genetic Engineering and Biotechnology | Akbari E.,Sharif University of Technology
Carbon | Year: 2015

In vivo dose-dependent effects of nanoscale graphene oxide (NGO) sheets on reproduction capability of Balb/C mice were investigated. Biodistribution study of the NGO sheets (intravenously injected into male mice at dose of ∼2000 μg/mL or 4 mg/kg of body weight) showed a high graphene uptake in testis. Hence, in vivo effects of the NGO sheets on important characteristics of spermatozoa (including their viability, morphology, kinetics, DNA damage and chromosomal aberration) were evaluated. Significant in vivo effects was found at the injected concentrations ≥200 μg/mL after (e.g., ∼45% reduction in sperm viability and motility at 2000 μg/mL). Observation of remarkable DNA fragmentations and chromosomal aberrations of the spermatozoa after ∼8 weeks from the first weekly injection were assigned to the involvement of the NGO in spermatogenesis of the mice. The uptake of the NGO in the testis could also increase the generation of reactive oxygen species in semen of the mice. Moreover, semen of the NGO-treated mice (containing the damaged spermatozoa) might disturb the hormone secretion and pregnant functionality of female mice (∼44, 35 and 59% reduction in fertility, gestation ability and multi-production capability) and also viability of the next generation (∼15% reduction in postnatal viability of delivered pups). © 2015 Elsevier Ltd.


Taha M.F.,Iran National Institute of Genetic Engineering and Biotechnology | Hedayati V.,Iran National Institute of Genetic Engineering and Biotechnology
Tissue and Cell | Year: 2010

Bone marrow and adipose tissue have provided two suitable sources of mesenchymal stem cells. Although previous studies have confirmed close similarities between bone marrow-derived stem cells (BM-MSCs) and adipose tissue-derived stem cells (ADSCs), the molecular phenotype of ADSCs is still poorly identified. In the present study, mouse ADSCs were isolated from the inguinal fat pad of 12-14 weeks old mice. Freshly isolated and three passaged ADSCs were analyzed for the expression of OCT4, Sca-1, c-kit and CD34 by RT-PCR. Three passaged ADSCs were analyzed by flow cytometry for the presence of CD11b, CD45, CD31, CD29 and CD44. Moreover, cardiogenic, adipogenic and neurogenic differentiation of ADSCs were induced in vitro. Freshly isolated ADSCs showed the expression of OCT4, Sca-1, c-kit and CD34, and two days cultured ADSCs were positively immunostained with anti-OCT4 monoclonal antibody. After three passages, the expression of OCT4, c-kit and CD34 eliminated, while the expression of Sca-1 showed a striking enhancement. These cells were identified positive for CD29 and CD44 markers, and they showed the lack of CD45 and CD31 expression. Three passaged ADSCs were differentiated to adipocyte-, cardiomyocyte- and neuron-like cells that were identified based on the positive staining with Sudan black, anti-cardiac troponin I antibody and anti-map-2 antibody, respectively. In conclusion, adipose tissue contains a stem cell population that seems to be a good multipotential cell candidate for the future cell replacement therapy. © 2010 Elsevier Ltd.


Akhavan O.,Sharif University of Technology | Ghaderi E.,Nanobiotechnology Research Laboratory | Hashemi E.,Iran National Institute of Genetic Engineering and Biotechnology | Rahighi R.,Sharif University of Technology
Nanoscale | Year: 2014

Graphene oxide nanoplatelets (GONPs) with extremely sharp edges (lateral dimensions ∼20-200 nm and thicknesses <2 nm) were applied in extraction of the overexpressed guanine synthesized in the cytoplasm of leukemia cells. The blood serums containing the extracted guanine were used in differential pulse voltammetry (DPV) with reduced graphene oxide nanowall (rGONW) electrodes to develop fast and ultra-sensitive electrochemical detection of leukemia cells at leukemia fractions (LFs) of ∼10-11 (as the lower detection limit). The stability of the DPV signals obtained by oxidation of the extracted guanine on the rGONWs was studied after 20 cycles. Without the guanine extraction, the DPV peaks relating to guanine oxidation of normal and abnormal cells overlapped at LFs <10-9, and consequently, the performance of rGONWs alone was limited at this level. As a benchmark, the DPV using glassy carbon electrodes was able to detect only LFs ∼ 10-2. The ultra-sensitivity obtained by this combination method (guanine extraction by GONPs and then guanine oxidation by rGONWs) is five orders of magnitude better than the sensitivity of the best current technologies (e.g., specific mutations by polymerase chain reaction) which not only are expensive, but also require a few days for diagnosis. This journal is © The Royal Society of Chemistry.


Lababpour A.,Iran National Institute of Genetic Engineering and Biotechnology
International Journal of Environmental Science and Technology | Year: 2016

The world is presently faced to the many calamities, mainly the increased and rapidly developing environmental changes, soil degradation as an example of such environmental problems which is correlated with the destructive effects of the sandstorms. Biological soil crust (BSC), a main component of soil, has various environmental functions including reduction in the erosion by increasing soil stability and providing a sanctuary for the growth of the taxa and vascular plants communities. Destruction of BSC, which naturally can be recovered slowly in a long time processes, contributes to the desertification and other environmental catastrophes. Therefore, accelerating the BSC recovery both the quality and the quantity of the crust development, especially in the desert areas, is of the prime interest. Recent advances in the BSC restoration have provided an immense potential for emulating the natural restoration methods mainly through providing soils with inoculant. This paper reviews the present restoration-based procedures for the biological soil crust restoration practice. The main landmarks are presented and highlighted including strain(s) selection and development, mass biomass production, inoculum preparation, soil inoculation, soil augmentation, nurseries, and crust succession monitoring and control. The review also introduces several successful case studies in the USA and the Republic of China. Thereafter, the paper briefly documents the future directions of the research and technologies. Development of a restoration system through the application of the microalgae inoculant is an encouraging aspect for accelerating the BSC recovery of the arid and semi-arid areas. However, further researches will help to establish and consolidate the potential of the microalgae cells and their application in desertification programs in large scales and in accordance with principles and requirements mandated by economic standards. © 2016, Islamic Azad University (IAU).

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