Showing 16 results for Hosseinkhani
Volume 1, Issue 3 (10-2014)
Abstract
Background: Despite the genetic heterogeneity reported in familial ALS (FALS), SOD1 gene mutations are the most frequent cause of FALS, accounting for around 20% of familial cases (ALS1) and isolated sporadic cases. Mutant forms of SOD1 exhibit toxicity that promotes the death of motor neurons. It is well documented that FALS produces protein aggregates in the motor neurons of FALS patients, which have been found to be associated to mitochondria.
Methods: In this study, we cloned the SOD1 gene, using reverse transcriptase-polymerase chain reaction (RT-PCR) method, from both a healthy control and a living 79 -year-old man with diagnosis of sporadic form of ALS who had shown unusual rapid progression of disease. RNA samples were available from lymphocytes of them. pET28a expression system and BL21 chemically competent Escherichia coli strain as host were used for protein expression.
Results: DNA Sequencing data showed both heterozygosis C to G transition at nucleotide position 21 leading to a C6W changing at protein level and a deletion at nucleotides position 73 to 169 leading to complete deletion of exon two.
Volume 9, Issue 2 (Spring 2021)
Abstract
Aims: Hospitalization in the cardiac care unit often imposes many physical and psychological tensions on the family. This study aimed to determine the effect of information support on depression of family caregivers of patients admitted to the cardiac intensive care unit.
Materials & Methods: This quasi-experimental study was performed on family caregivers of patients admitted to the cardiac care unit of Sevom Shaban Hospital of Damavand city, Iran, 2020. Participants included 60 family caregivers who were assigned to intervention and control groups. Family caregivers in the intervention group received information support through a training booklet, and the control group received the same routine information. Data collections were conducted by a demographic data sheet and anxiety and depression questionnaire (HADS). The data were then analyzed using SPSS 18 software.
Findings: The two family groups (intervention and control) showed different levels of depression after the information support intervention (p=0.02). Wilcoxon's signed-rank test showed that the mean depression of family caregivers after the intervention was 4.63±2.67 compared to the mean depression of 6.50±3.01 in the control group (p<0.05).
Conclusion: During the patient's stay in the cardiac care unit, nurses can alleviate family depression by providing information support.
Volume 9, Issue 3 (Summer 2018)
Abstract
Aims: The probability of establishing electrostatic interactions due to the abundance of charged hydrophilic residues and especially arginine is considered the most important thermal stabilizing factor of thermophilic enzymes. The current study was conducted with the aim of comparing thermodynamic stability and kinetic refolding of Lampyris turkestanicus and some of its mutants.
Materials and Methods: In the present experimental thermal stability and the way of refolding Lampyris turkestanicus and 3 mutations, including ERR, ERR/I232R, ERR/Q35R/I182R/I232R were investigated by various spectroscopic techniques. In order to high expression of proteins, a single clone of each sample was selected and inoculated into 10ml of LB culture medium, containing Kanamycin at a concentration of 50μg/mg and incubated at 37°C with an ideal aeration for 12-15 hours. The culture medium was centrifuged for 5 minutes at 5000g at 4°C to provide the cellular contents of the bacteria. The results were obtained through spectroscopic methods of remote and near circular dichroism, intrinsic fluorescence, differential scanning calorimetry, and kinetics experiments, using fluorescence-stopped flow technique.
Findings: Along with the increase in the number of arginine residues at the protein level, the stability and structural compression of the mutated enzymes in comparison with the wild enzyme were increased and the thermograms obtained from differential scanning calorimetry showed a slight increase in Tm and calorimetric enthalpy of mutated proteins in comparison with wild protein.
Conclusion: The rate constant of refolding mutated enzymes has increased compared with the wild type. The improvement of thermodynamic and kinetic parameters results from the improvement of electrostatic interactions, which results in a higher degree of compression and structural density.
Volume 10, Issue 1 (Winter 2019)
Abstract
One of the main challenges in the treatment of genetic disorders, such as cancer, is of drug delivery systems and their inability to monitor and track delivered drug to the targeted site. Therefore, the design of novel with dual capabilities of nuclear drug delivery and tracking into a research priority for this field’s The aim of this study is to design based on both non-cytotoxic quantum dots and chimeric peptides, with dual tracking and delivering small genetic agents into the nucleus. The GQDs with green emission color were synthesized by Hummer’s and methods and characterized by UV-Vis, photoluminescence (PL), Raman spectroscopies, and scanning electron microscopy (SEM). conjugated with MPG-2H1 chimeric peptides through noncovalent interactions. Following conjugation step, the ζ-potential of the complex increased (From -38.6 to -11.1 in complex1, -9.6 in complex2 and -5.74 in complex3). The conjugation was confirmed by native acrylamide gel retardation assay. The of the GQDs was investigated by MTT assay and finally, was carried out. The results showed that MPG-2H1/ GQD complexes can enter cells; however, free-GQDs didn’t enter the cells significantly.
Volume 10, Issue 2 (Spring 2019)
Abstract
Luciferase from firefly Photinus pyralis (P .py) is a peroxisomal enzyme that converts a heterocyclic substrate luciferin to an excited state oxyluciferin in the presence of Mg+2-ATP and O2. Excited oxyluciferin with the emission of visible light is changed to its ground state. The combination of rapidity, sensitivity, and convenience has led to the development of a broad range of luminescence applications. In spite of wide ranges applications, firefly luciferase is unstable against changes in chemical and physical conditions, thereby reduce its precision and sensitivity. The most undesirable instability of the luciferase is low thermostability and high susceptibility to proteolytic degradation. According to previous studies, limited proteolysis by trypsin of P .py luciferase indicated six cleavage sites on two accessible regions: 206-220 (Including K206, R213, and R218) and 329-341 (Including K329, R330, and R337) on N-terminal domain. In this study, we used site-directed mutagenesis to introduce one point mutation on the 329-341 accessible regions of P. py luciferase, in order to investigate the role of R330 on the enzyme structure and function which R330 changed to Q. Based on limited proteolysis data, R330Q mutant didn’t significantly change compared to wild type, but this mutation caused several alterations in enzymatic properties including shifting the pH optimum from 7.5 to 8 and increasing the thermal inactivation. Based on the results, it can be concluded that whilst Arg330 is a conserved residue but not effects on trypsinolysis stability.
Volume 10, Issue 4 (Fall 2019)
Abstract
The SPTBN4 gene, a part of the spectrin protein family, plays important roles in various cellular processes, including cell cycle, nerve cell development, and so on. Recently, a new miRNA has been found in this SPTBN4 gene, which was registered at the NCBI database. The aim of the present study was to investigate the expression of this miRNA, called SPTBN4-miR1, in the process of differentiation of human embryonal carcinoma cell line NT2 and also the overexpression effect of this miRNA on the differentiation of these cells. RT-qPCR results indicate that SPTBN4-miR1-5p and SPTBN4-miR1-3p show a significant increase in expression in the process of neural differentiation from day three until the 8th and 14th day of differentiation. Then, after overexpressing the SPTBN4-miR1 precursor in NT2 cells and retinoic acid treatment, the expression of pluripotent and differentiation revealed the role of SPTBN4-miR1-5p and SPTBN4-miR1-3p in promoting differentiation and exclusion from the pluripotent state. It seems that by making further studies and finding out the possible targets of these miRNAs, a distinctive marker can be achieved and used to improve the differentiation process.
Volume 11, Issue 2 (Spring 2020)
Abstract
The bioluminescence process is a widespread phenomenon in Nature. These enzymes are identified in some domains of life, but the luciferases from the lampyridea genus are considered for biological applications. The molecular cloning of a new type of Iranian firefly luciferase from Lampyroidea maculata was reported, previously. In this study, we analyzed the rare codons of the Iranian insect luciferase gene using the computational databases as ATGme, RACC, LaTcOm, and Sherlocc. Also, the structural modeling process of this enzyme was performed. Next, the status of these rare codons in this structural model was studied using SPDBV and PyMOL software. In the following, the substrate binding site was studied using the AutoDock Vina. By molecular modeling, some rare codons were identified that may have a critical role in the structure and function of this luciferase. AutoDock Vina was used in the molecular docking that recognizes Asp531 that yield closely related to luciferin and AMP binding site.. This bioinformatics analyzes play an important role in the design of new drugs.
Volume 11, Issue 3 (Summer 2020)
Abstract
Inhibitor of apoptosis (IAP) are a family of proteins that block cell death through caspase activity. Survivin is smallest IAPs family member that overexpresses in different cancer types but not in normal tissue except embryonic tissue. Survivin may be used as a new marker to stratify cancer patients for more optimal treatment modalities. The aim of the current study was to investigate survivin DNA cloning into pET-28a and its expression in E.coli.
The sequence of survivin gene was amplified by PCR using specific primers and pcDNA-survivin temple. PCR product and pET-28a plasmid were digested by HindIII/NheI restriction enzymes and survivin was ligated into the digested vector. Then, the ligation product was transformed into the E.coli DH5a competent cells and screened by antibiotic selection marker (kanamycin). Positive colonies were selected by colony PCR and screened by double digestion of isolated plasmid. One positive colony was sequenced and confirmed. The recombinant plasmid was transformed into the expression strain of E.coli (BL21) by chemical method. The expression of survivin was induced in the different conditions and expression level investigated by SDS-PAGE.
The size of PCR product in agarose gel showed the correctness of amplification. The digested pET-28a plasmid also indicated the correctness of enzymatic reaction. The sequence of the cloned fragment revealed a 100% similarity to the human survivin. In expressing, adding IPTG increased the expression of survivin protein in all conditions, especially 37 ᵒC from 2 h after induction. At all conditions, most of survivin accumulated in the bacteria as inclusion body.
Volume 11, Issue 4 (fall 2020)
Abstract
Developing a technique for efficient and safe gene delivery to plant cells is a fundamental aim of plant biotechnology. Agrobacterium mediated transformation as the most common and practical method in plant gene delivery has considerable difficulties such as limitation in applicable for plant species. In recent years several new methods have been suggested, although none of them could be a good replacement. The use of nanotechnology has been provided new solutions to overcome the limitations of biotechnology. Designing biocompatible nanostructures for passing cell barriers and targeted delivery of cargo has improved the biological achievements. In this research the capability of arginine functionalized single-walled carbon nanotube (Arg-SWNT) as a new carrier to transfer plasmid DNA, which codes green fluorescent protein (GFP) to tobacco suspension cells, has been investigation. It is suggested that single-walled carbon nanotubes can pass through cell wall pores and plasma membrane while it carries plasmid DNA along with. The fluorescence microscopy images illustrate the success of gene delivery by Arg-SWNT
Volume 13, Issue 3 (1-2023)
Abstract
Aims: Programmed cell death is a vital cellular process that is highly conserved in evolution. Apoptosis, as a common mode of programmed cell death, is disturbed in the most human malignancies and leads the resistance of cancers to current treatment strategies. Caspase 9 is a key protein in mitochondrial apoptosis. Activated Caspase 9 leads to activation of Caspase 3/7, initiating a caspase cascade and killing cell. In this study, Caspase 9 gene was cloned into pcDNA3.1(+) and its expression and function evaluated in cell.
Methods: PCR amplification of Caspase 9 was performed by specific primers and ligated into pcDNA3.1(+) after double-digestion with KpnI and BamHI. After sequencing, pcDNA/Caspase 9 was transfected into SH-SY5Y cells and treated with doxorubicin. Caspase 9 function was determined by its effect on cell death level by trypan blue and PI staining, and Caspase 3 activity, and its expression in cells measured by western blotting.
Finding: Caspase 9 gene cloning was done and its expression in cell defined by western blot. Overexpression of Caspase 9 led to autoprocessing following homodimerization and induction of cell death and also increased cell sensitivity to doxorubicin treatment and declined cell viability.
Conclusion: The cloned Caspase 9 was functional in cell and enhanced apoptosis in the treated cells by doxorubicin through self-activation and subsequently amplification of Caspase 3 activation.
Volume 14, Issue 2 (6-2011)
Abstract
Objective: Evaluating the effects of p-benzoquinone and hydroquinone on the RUNX2 expression and osteoblastic differentiation of human marrow derived mesenchymal stem cells (MSCs).
Materials and Methods: Bone marrow MSCs obtained by cultivating marrow mononuclear cells, were exposed to 10μM of either p-benzoquinone or hydroquinone. Following chemical treatment, RUNX2 gene expression was assessed by Real-time RT PCR 1, 6, 24 and 48 hours later and osteogenic differentiation was analyzed using alizarin red and alkaline phosphatase staining methods on days 7 and 14 after ostegenic induction.
Results: RUNX2 expression was significantly elevated (up to approximately 8 times) due to chemical exposure but the applied chemicals exert no considerable effect on MSCs osteogenic differentiation.
Conclusion: According to the literature, despite the necessity of RUNX2 overexpression on the induction of osteogenic differentiation, but it is not sufficient for osteogenesis to occure so increase in RUNX2 expression observed in our study is not the indicator of the induced osteogenic differentiation. Instead, this elevated expression could be the sign of increased activity of the canonical Wnt signaling pathway thereby its involvement in the development of AML due to exposure to benzene and its metabolites. Moreover, this augmented expression of RUNX2 in MSCs can indicate the
RUNX2 overexpression in myeloid progenitors as an expected similar effect of exposure to benzene and its metabolites to contribute in myeloid malignancies developed due to benzene exposure.
Volume 14, Issue 4 (9-2023)
Abstract
SH-SY5Y is a neuroblastoma cell line which used as a cancer and neurodegenerative disorders model and its neuro-experimental studies. The different diseases cause by a defect in apoptosis pathway. Disruption of apoptotic proteins has an effect on the treatment process and response to drugs. In nerve cells, due to the high expression of apoptosis inhibitory proteins, the efficacy of drugs is low. Combination therapy is one of the developing treatment methods. The aim of this research is to evaluate the effectiveness of doxorubicin drug on apoptosis in SH-SY5Y cells under the conditions of high expression of caspase9. Caspase9 is a key enzyme in intrinsic apoptosis. First, cell viability was obtained through MTT assay under the different drug concentrations. Then, caspase9 gene was transfected in cells and affected by the concentration lower than IC50 of drug, and cell energy level and cell death were checked by different methods. ATP assay showed that the expression of caspase9 with drug lead to ATP decreases. Caspase3/7 activity indicated an increase in cell death by drug and caspase. Propidium staining to hoechst showed that the expression of caspase9 in combination with doxorubicin induce more death. To ensure the expression levels of protein that induces cell death, the amount of caspase3 protein was checked by western blotting, which showed a significant increase in combination of caspase9 and drug. Our findings showed that the induction of caspase9 expression intensifies the effect of drug and the combined treatment may be effective on the responsiveness of neuronal diseases.
Volume 15, Issue 2 (5-2024)
Abstract
Gene delivery using the force of a magnetic field is called magnetofection. The purpose of this study is the synthesis and characterization of magnetic iron oxide nanoparticles (Fe3O4) as the core of the transfer agent and to investigate the effect of alternating magnetic field on transfection efficiency. For this purpose, the first magnetic nanoparticles (MNP) were synthesized by coprecipitation method. The magnetic properties of the synthesized MNP were investigated by vibrating sample magnetometer (VSM), appearance characteristics, and zeta potential of the synthesized particles were evaluated using transmission electron microscopy (TEM) and dynamic light scattering (DLS). Then, using magnetic nanoparticles (MNP), polyethylene imine (PEI) and plasmid DNA containing luciferase reporter gene (pDNA), PEI-pDNA binary complex and MNP-PEI-pDNA ternary complex were synthesized. The complexes were evaluated using DLS and gel retardation techniques. The results of DLS and gel retardation technique showed that the complexes have a suitable surface charge and polyethyleneimine is well joined to pDNA and neutralized its negative charge. Finaly, human breast cancer cell lines (MCF-7) and Hek293T cells were transfected by ternary complex in the presence of 50 Hz alternating magnetic field. Cell viability was measured using the MTT test. The obtained results showed that the transfection efficiency in the cells that were transfected with the ternary complex in the presence of alternating magnetic field increased significantly compared to the control group, without any additional toxicity (P ≤ 0.05).
Volume 16, Issue 2 (9-2013)
Abstract
Objective: In vitro ovarian follicle culture provides a tool to investigate folliculogenesis and may be used as an assisted reproductive technology )ART(. This study aims to compare survival and development rates in mouse ovarian follicles after two and three dimensional in vitro cultures.
Methods: Preantral follicles were isolated from the ovaries of 14-day old female mice and cultured in α-MEM medium supplemented with 5% FBS for 12 days in a two dimensional and three dimensional culture with different concentrations of sodium alginate (0.25%, 0. 5%, or 1%). The follicle diameter, survival and maturation rate during culture were analyzed and compared with one-way analysis of variance (ANOVA). P<0.05 was considered as statistically significant.
Results: The mean diameter of preantral follicles that capsulated with 0.5% alginate was significantly higher than other concentrations in each group on days 6 and 12 (P<0.001). The percentages of follicles which released metaphase II (MII) oocytes in the two dimensional groups and in the three dimensional groups at 0.25%, 0.5% and 1% concentrations of sodium alginate were 29.03%, 33.33%, 44.18% and 35.89% respectively. The percentage of MII oocytes was significantly higher at the 0.5% concentration of sodium alginate (P<0.001).
Conclusion: Follicles encapsulated in 0. 5% sodium alginate in three dimensional culture displayed the highest survival development and maturation rate.
Ebrahim Jahanshahi, Ali Hosseinkhani, Seyed Mohammad Hojat Moahmmadi,
Volume 16, Issue 12 (2-2017)
Abstract
Solar humidification-dehumidification desalination is one of the most practical methods for water desalination in small scale for regions far from cities and low population. The aim of this study is manufacturing and simulation of a solar humidification-dehumidification desalination system with capacity of 20 lit/day. This system consists of humidification and dehumidification units, solar air and water heaters. To this end, at first this system is explained and modeled. Then, manufacturing process of solar air heaters, and different parts of desalination system is investigated. After the manufacturing process of the desalination system, this system is experimentally tested and the effect of pertinent parameters, such as the temperature of inlet water and air to humidifier; inlet water temperature and flow rate to dehumidifier on the performance of the system and distillate product is investigated. The results show that the effect of water temperature on the fresh water produced in more than air temperature. Moreover, using the chilled water, which its temperature in the range of well temperature, in the dehumidifier inlet leads to an increase of 31 % in the fresh water produced. Also, the best water flow rate to the dehumidifier inlet is 0.12 kg/s. Finally, experimental and simulation results are compared with each other and good consistency is seen.
Mehdi Rostami Hosseinkhani1, Pourya Omidvar, Sara Allahyaribeik, Masoud Torabi Azad,
Volume 18, Issue 3 (5-2018)
Abstract
Dispersion of oil pollutants is one of the important topics of great concern which should be modeled for a wide range of hydrodynamic systems such as seas and oceans. In this paper, the effects of using booms on the oil plume are simulated using the Smoothed Particle Hydrodynamics (SPH) Method. The open-source SPHysics2D code is developed into two phase by adding the effects of surface tension and an added pressure term to the momentum equation. Several problems of plume dynamics are shown, and the performance of the developed code is evaluated. Firstly, the rising pattern of an oil plume with the density ratio of 0.8 is simulated where the results are compared with the analytical solution. Then, the rising pattern of a plume with density ratio of 0.1 is simulated and the time evolutions of the rising velocity and center of mass are shown. The simulation of the cnoidal wave on beaches is conducted and compared with an available experimental result. Finally, the effects of a boom with different angles on the oil plume dispersion are investigated. It will be shown that the SPH method could be an optimized method for the numerical simulation of the complex problems such as water wave dynamics and two-phase flows.
