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TGF-b Downregulation by RNAi Technique in ex Vivo-Expanded HSCs on 3D DBM Scaffold
ZS Hashemi,M Forouzandeh Moghadam,M Soleimani,M Hafizi
Tehran University Medical Journal , 2012,
Abstract: Background: Bone Marrow Transplantations (BMT) are limited by low CD34+ cell counts in umbilical cord blood (UCB) and these cells need to be expanded for success in such procedures. To achieve this goal, ex vivo expansion of hematopoietic stem cells (HSCs) by enhancing their self-renewal activity on demineralized bone matrix (DBM) scaffold coated with mesenchymal progenitor cells (MPCs) and unrestricted somatic stem cells (USSCs) was recommended. TGF-b pathway is a key inhibitory factor for HSCs self-renewal. In this study ex vivo expansion and downregulation of TGF-b pathway were simultaneously performed. Methods: USSC cells were isolated from UCB and then coated on DBM scaffold as a feeder layer. UCB CD34+ cells were isolated from UCB by magnetic activated cell sorting (MACS) method and were transfected by siRNA against TGFbR2 in two-dimensional (2D) and three-dimensional (3D) cultures by co-cultivation with USSC. TGFbR2 expression levels were evaluated by quantitative real-time PCR. Cell count and flow cytometry were performed and clonogenic activity was evaluated. Results: Ex vivo expansion of CD34+ cells was significantly enhanced (41±0.7 folds) by TGFbR2 downregulation, especially in 2D than 3D cultures. Finally, 2D culture showed less TGFbR2 expression levels and higher increase in the percentage of CD34 markers by flow cytometry assay. Conclusion: The 3D siRNA delivery system would be of lower efficiency in contrast to 2D settings where the cells have less freedom and are in more contact with the feeder layer.
Comparing MALDI-TOF Mass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus Faecalis Isolated from Clinical
Samadi Kafil,H.,Mohebati Mobarez,A,Forouzandeh Moghadam,M.
Medical Laboratory Journal , 2013, DOI: http://www.goums.ac.ir/mljgoums/index.php?&slct_pg_id=10&sid=1&slc_lang=en
Abstract: Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora, in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of MALDI-TOF mass spectroscopy method through Biochemical and Molecular methods for detecting Enterococcus faecalis and Enterococcus faecium.Materials and Methods: seventhy five clinical samples were collected for biochemical, molecular and mass spectroscopy investigations. Samples were treated with Esculin hydrolysis, Catalase, Pyrrolidonyl aminopeptidase, 6.5% NaCl solution, motility, 0.04% Tellurite, L-Arabinose and Sorbitol. Using specific primes allele specific PCR was used.The samples were then analyzed by MALDI-TOF mass spectroscopy and Biotyper 3 software.Results: Enterococcus faecium and Enterococcus faecalis were detected in thirty and forty two samples, respectively whereas three samples showed both bacterial infections. Using biochemical analysis, two E. faecium isolates were Arabinose negative and one E. faecalis isolates was Telliurite negative. All samples were showed correct bands in PCR results but two of them didn't show clear bands(on agarose gel). In mass spectroscopy analysis all strains were correctly detected and well defined.Conclusion: According to our results, MALDI-TOF mass spectrometry in comparison with Molecular and Biochemical Methods could be a reliable and accurate method that can easily and quickly identify and differentiate Enterococcus faecium and Enterococcus faecalis in clinical samples.Key words: Enterococcus faecalis, Enterococcus faecium, MALDI-TOF mass spectrometry, PCR
Design and development of an in-house multiplex RT-PCR assay for simultaneous detection of HIV-1 and HCV in plasma samples
M Paryan,M Forouzandeh Moghadam,V Kia,S Mohammadi-Yeganeh
Iranian Journal of Microbiology , 2012,
Abstract: Background and Objectives: HIV-1 and HCV infections are life threatening problems in patients who receive blood products. Serological methods have proven useful in detecting these infections, but there are setbacks that make it challenging to detect these infectious agents. By the advent of Nucleic Acid Testing (NAT) methods, especially in multiplex format, more precise detection is possible.Materials and Methods: We have developed a multiplex RT-PCR assay for simultaneous detection of HIV-1 and HCV. Primers were designed for highly conserved region of genome of each virus. Using these primers and standard plasmids, we determined the limit of detection, clinical and analytical specificity and sensitivity of the assay. Monoplex and multiplex RT-PCR were performed.Results: Analytical sensitivity was considered to be 100 and 200 copies/ml for HIV-1 and HCV, respectively. High concentration of one virus had no significant effect on the detection of the other one with low concentration. By analysis of 40 samples, clinical sensitivity of the assay was determined to be 97.5%. Using different viral and human genome samples, the specificity of the assay was evaluated to be 100%.Conclusions: The aim of this study was to develop a reliable, rapid and cost effective method to detect HIV-1 and HCV simultaneously. Results showed that this simple and rapid method is perfectly capable of detecting two viruses in clinical samples.
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2)
M Mahami-Oskouei,A Dalimi,M Forouzandeh-Moghadam,MB Rokni
Iranian Journal of Parasitology , 2011,
Abstract: Background: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species.Methods: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and ana-lyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifi-cally of Fasciola species. Results: The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates.Conclusion: The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates.
Effects of Treatment with Bone Morphogenetic Protein 4 and Co-culture on Expression of Piwil2 Gene in Mouse Differentiated Embryonic Stem Cells
Zohreh Makoolati,Mansoureh Movahedin,Mehdi Forouzandeh-Moghadam
International Journal of Fertility & Sterility , 2009,
Abstract: Background: Specific growth factors and feeder layers seem to have important roles in in vitroembryonic stem cells (ESCs) differentiation. In this study,the effects of bone morphogenetic protein4 (BMP4) and mouse embryonic fibroblasts (MEFs) co-culture system on germ cell differentiationfrom mouse ESCs were studied.Materials and Methods: Cell suspension was prepared from one-day-old embryoid body (EB)and cultured for four days in DMEM medium containing 20% fetal bovine serum (FBS) in thefollowing groups: simple culture (SC), simple culture with BMP4 (SCB), co-culture (CO-C) andco-culture with BMP4 (CO-CB). Expression of piwi-like homolog 2 (Piwil2), the germ cell-specificgene, was evaluated in the different study groups by using quantitative real time polymerase chainreaction (RT-PCR). Testis was used as a positive control.Results: The maximum and minimum Piwil2 expression was observed in SC and SCB groups,respectively. A significant difference was observed in Piwil2 expression between SCB and otherstudy groups (p<0.05).Conclusion: The findings of this study showed that neither the addition of BMP4 in culture mediumnor the use of MEFs as a feeder layer have a positive effect on late germ cell induction from mouseESCs.
Effect of losartan on NOX2 transcription following acute myocardial ischemia-reperfusion
Fatemeh Safari,Sohrab Hajiadeh,Seyed Hosein Moshtaghion,Mehdi Forouzandeh Moghadam
Physiology and Pharmacology , 2012,
Abstract: Introduction: Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-2 (Nox2) is one of the predominant sources of ROS production during myocardial ischemia-reperfusion and can be induced by angiotensin II. The evidence suggests that pharmacological blockers of renin-angiotensin system can exert direct tissue effects independent of their ability to regulate blood pressure. The mechanism(s) responsible for such direct effects are not well understood. The aim of this study was to investigate the early changes of cardiac NOX2 gene transcription after myocardial ischemiareperfusion in rats treated with losartan, an angiotensin type 1 (AT1) receptor blocker. Methods: Wistar rats were divided into five groups: Control, sham operated, ischemia-reperfusion (group IR), losartan without ischemia and losartan with ischemia-reperfusion. The animals underwent 30 min of left anterior descending artery occlusion and subsequent reperfusion for 180 min. The mRNA expression was determined by real time-PCR in ischemic area of the left ventricle (LV) and non ischemic area of right ventricle (RV). Results: Compared to control hearts, exposure to myocardial ischemia-reperfusion produced a significant increase in NOX2 mRNA level in ischemic area of LV (P<0.001) but not in non ischemic area of right ventricle. Although in losartan group, NOX2 mRNA levels neither in LV nor RV were significantly altered, while in losartan and ischemiareperfusion group NOX2 mRNA upregulation in ischemic area was significantly suppressed (P<0.01). Conclusion: Based on the obtained results, it could be concluded that following acute myocardial ischemia– reperfusion, NOX2 mRNA levels were increased in ischemic area of left ventricle but not in non ischemic area of right ventricle, suggesting the local effect of ischemia on the gene expression. Furthermore, inhibition of NOX2 transcription in ischemic area may be a mechanism of the anti ischemia effects of losartan.
The Effect of Bone Morphogenetic Protein 4 on the Differentiation of Mouse Embryonic Stem Cell to Erythroid Lineage in Serum Free and Serum Supplemented Media
Mohammad Ali Owchi,Mojdeh Salehnia,Mehdi Forouzandeh Moghadam,Mandana Beigi Boroujeni
International Journal of Biomedical Science , 2009,
Abstract: This study was done to compare the effects of bone morphogenetic protein-4 (BMP-4) on mouse embryonic stem cells (ESC) differentiation to erythroid lineage in serum free and serum supplemented media. The embryoid bodies (EBs) cells were seeded in semisolid serum free and serum supplemented media in the presence of different concentrations of BMP-4. The erythroid colonies were assessed morphologically, ultrastructurally and by benzidine staining. The expression of the epsilon (ε), βH1 and βmajor globins, Runx1 and β2m genes was evaluated by Real time PCR. The colony size and the percent of benzidine-positive colonies increased in both BMP-4 supplementd groups but the number of colonies were lower in these groups than control. Erythropoiesis related genes were expressed in both serum free and serum supplemented groups. There were not significant differences between the ratios of genes expression to β2m in these groups except the ratio of Runx1 was significantly higher in serum free group (P<0.05). The ratio of ε and βH1 to β2m in EBs was higher than both BMP-4 containing groups (P<0.05) and βmajor was not expressed in EB cells. These findings showed in serum free condition the effects of BMP-4 on the erythroid differentiation was prominent than serum supplemented group.
Dissipative chaos in semiconductor superlattices
F. Moghadam,M. Esmaeilzadeh
Iranian Journal of Physics Research , 2008,
Abstract: In this paper the motion of electron in a miniband of a semiconductor superlattice (SSL) under the influence of external electric and magnetic fields is investigated. The electric field is applied in a direction perpendicular to the layers of the semiconductor superlattice, and the magnetic field is applied in different direction Numerical calculations show conditions led to the possibility of chaotic behaviors.
Accounting and comparing of expenditure on the medical services given at neuro-surgery department of Imam Khomeyni Hospital in the year of 1994 (1373)
Abasi Moghadam M
Tehran University Medical Journal , 1998,
Abstract: This study was focused on analysis of expenditure on all the medical services given at Neuro-Surgery Department of Imam-Khomeini Hospital in the year of 1994 (1373). In this study, all the information on descriptive method and the techniques of cost analysis and cost per unit of service provided accountancy, were analysed. 573 patients were considered in this study. 522 of them underwent 13 different types of neuro-surgery operations. 92.6% of them total departmental costs were related to current expenditures and 7.4% of that was related to the capial expenditures. The personnel costs with 49% was the highest portion of the total costs. Percentage wise, the costs were as follows: Medicine, materials and equipment 22%, food 17.6%, depreciation 7.4%, fuel, water, electricity and telephone 3.5%. The mean duration of stay was 16.3 days for every in-patient. The percentage of occupied bed was 58% if the percentage of desired bed occupancy was supposed 80%, therefore, 22% of the bed, plus 3512 bed-day were gone wasted. The real cost of med-care policy need to be more rational for the operation and hospitalization. It should be mentioned that the wasted time was 886 hours and wasted cost was 71, 708, 410 Rials in operation room.
Statistical study and review of prostatic latent carcinoma
Jamali M,Moghadam K
Tehran University Medical Journal , 1996,
Abstract: The autopsies, which have been performed within the last 50 years, have revealed that real prevalence of prostatic carcinoma is more frequent than clinical one. The real prevalence of prostatic carcinoma, is prevalence combination of carcinomas which have been revealed clinically (They have been confirmed by autopsy or by operation) and the prostatic latent carcinomas are those, which are found in autopsy or randomly in the biopsies taken for hyperplasia. But they have no clinical syndromes. In order to review prevalence of prostatic latent carcinoma in Iran, all prostatic lesions (Including hyperplasia or carcinoma) were studied in Imam Khomeini medical complex during 10 years (1981-91), in university Jihad center and medical center of Iran within 2 years and in Yazd faculty of medicine within 3 years (1981-84). The total cases were 1110 among which 1085 cases were selected upon reviewing for statistical analysis. At first all lamellas were studied, then the ratio of adenocarcinoma to total prostatic lesions were analyzed and types of carcinoma and their percentage in total cases were identified. Finally the prostatic latent carcinoma and its percentage in total malignancy cases were presented
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