M Habibi Roudkenar, M Oloomi, M.a Shokrgozar, N Shahrokhi, A Mohammadi Roushandeh, Y Kuwahara, M Fukumoto,
Volume 3, Issue 1 (Spring 2006)
Abstract
Abstract
Background and Objectives
Immunotoxins are comprised of cell targeting and cell killing moieties. Immunotoxins have been proposed as new strategy for cancer treatment. In this study, catalytic domain of Shiga-toxin (A1) was fused to human Granulocyte Macrophage-Colony Stimulating Factor (h GM-CSF) . The fused gene was already expressed in E.coli and the expressed protein was analysed for its cytotoxic activity on human cancer cell lines expressing GM-CSF receptor (GM-CSFR). Moreover, the impact of GM-CSF receptor on inhibition of cytotoxic effect of Shiga-toxin-GM-CSF recombinant hybrid in cell lines expressing GM-CSF receptor (GM-CSFR) was evaluated.
Materials and Methods
Cell lines were grown in RPMI-1640 medium supplemented with 10-20% heat inactivated fetal bovine serum (Gibco-BRL). Cytotoxic activity was checked on the cell lines and was measured by trypan blue staininig and MTT assay. GM-CSF receptor was blocked by anti-GM-CSF antibody.
Results
Cytotoxicity studies revealed that the chimeric protein induced cytotoxic effect on different cell lines. This effect was found to be specific due to the presence of killing moiety (A1) that exerts its effect through specific cell targeting domain i.e. GM-CSF, by binding to its receptor present on those cell lines. K562 and LS174T were the most sensitive .
Conclusions
Our results revealed that the hybrid protein is toxic to the cancer cell lines bearing GM-CSF receptor this effect could be a potential factor for further consideration of hybrid protein as a therapeutic agent.
Key words: Shiga-toxin , Granulocyte macrophage - colony stimulating factor ( GM–CSF ) , Cytotoxicity, Cancer cells
H Khorsand Mohammad Pour, I Nourmohammadi, M.a Jalili, Z Motallebi,
Volume 3, Issue 1 (Spring 2006)
Abstract
Abstract
Background and Objectives
Albumin is currently used in greater volume than any other biopharmaceutical solution that is available. The most large- scale production of human albumin is still conducted by cohn cold ethanol fractionation (Cohn's method). In order to save the operating time, labor and material in manufacturing albumin 20%, a two-stage process was carried out based on the cold ethanol fractionation method.
Materials and Methods
For this experimental study, fresh frozen plasma (FFP) was used as a starting material. In the first stage by addition of supernatant IV to a dissolved fraction V paste, a dense fraction V paste (Vd+I) containing the highest possible albumin concentration and the least impurity (ethanol, a globulins and ionic concentration) was recovered by centrifugation the recovered paste was then dissolved and filtered through a depth filter in order to remove a globulins as impurity. The filtrate (HPs) was suitable to produce albumin 20%. Two batches were produced and all the methods for the evaluation of the finished product were applied according to the guidelines of the British Pharmacopoeia.
Results
The quality control of the albumin 20% as a finished product from 2 batches (batches 1 and 2) in the final container produced by the proposed procedure led to satisfactory results. Appearance of the product was clear, purity over 99% , and the molecular size distribution of albumin was revealed for aggregates or polymers less than 2.7 % . The yield of albumin was 24 ± 1 gr/kg of plasma.
Conclusions
Advantages of the proposed process as compared with the rountine process (Cohn & Kistler- Nitschmann methods) during the production of intermediate products (Vd+I, HPs) were remarkable savings in terms of operating time, material and manpower at about 50%.
Key words: Human serum albumin, Plasma fractionation, Cohn Fraction V
B Damari, S Torabian, A Gharehbaghian, M Magsudlu, N Mohammadi, M Naser Bakht, M Rahbari, E Sanei Moghadam, A.h Asadi, M Paridar,
Volume 3, Issue 2 (Summer 2006)
Abstract
Abstract
Background and Objectives
One of the main objectives of blood centers is to promote voluntary blood donation. It is important then to recognize people views and beliefs about blood donation. Decrease or elimination of blood replacement is closely correlated with the tendency of people to embark on voluntary blood donation. In recent years, IBTO has taken measures to eliminete blood replacement and it has been considered as a priority in the strategic planing of IBTO. Blood replacement rate in most of the provinces is reported to be zero, except in 3 provinces of Hormozgon, Sistan and Baluchestan, and Khuzestan. A qualitative study was done considering the role of views and beliefs of people and their tendency to embark on blood donation in 3 southern provinces which are faced with problems regarding blood donation.
Materials and Methods
Two focus group meetings were held in each of these provinces. In each session, various and specified groups of people participated with pre-determined standards.
Results
False beliefs and views about blood donation were brought up: transmission of infectious diseases to donors, blood deficit being an improper claim, blood donation just for men, girls being banned to donate blood, blood donation making one thin and revealing addiction, phlebotomy better than blood donation, religious prohibition of blood donation, and the negative effect of blood donation on sexual behaviors.
Conclusions
In order to promote the motivation of people of these regions to appreciate voluntary blood donation, besides using social marketing tools, it is necessary to use these beliefs and views in social marketing and provincial publicity. The false views which are common among these three provinces can be rectified through similar programs and those specific to each province can be addressed by a different strategy.
Key words : Blood donor , Qualitative research , Social marketing
Ladan Hosseini Gohari, Isa Noormohammadi, Ali Akbar Sharafi Tafresi Moghadam, Leila Mostaan, Aneti Drousiotou,
Volume 3, Issue 4 (Winter 2007)
Abstract
Abstract
Background and Objectives
For a successful prevention program, globin chain synthesis, as a complementary test beside DNA analysis, is necessary. So, it is important that each laboratory establishes its own reference range for the classification of thalassemia syndromes by globin chain synthesis. Globin chain synthesis is a relatively complex test introduced in the study of thalassemia syndromes as a reference method. The technique is also useful for variant chain identification.This study aims to stablish the method of globin chain synthesis to determine a / b chain ratio in healthy individuals.
Materials and Methods
In this study globin chain analysis was performed on 30 healthy laboratory personnels with normal HbA2 and normal hematological indices. In this method a reticulocyte-rich sample is incubated with a mixture of amino acids, one of which (leucine) radioactively labelled. After washing the excess radioactivity and precipitating the globin, different chains are separated by cation exchange chromatography.
Results
The mean a / b ratio was 1.045 ± 0.12 (mean ± 1SD) in healthy subjects. Our findings were in agreement with those of the other investigators in the world.
Conclusions
In any screening program, diagnostic problems will arise that can not be solved without biosynthetic studies. The Clegg and Weatherall method has been proved to be very reliable and reproducible, but time-consuming (requiring four days). New methods like reverse phase HPLC are now available for chain separation. Therefore, according to the procedures each laboratory should determine its own reference range.
Key words : a globin, b globin, Hemoglobin, Normal range, Ion exchange chromatography
M. Allah Bakhshian, M.h. Mohammadi, Gh. Rastegar Lari, A. Kazemi, F. Ala, Sh. Ravanbod, A. Allah Bakhshian,
Volume 5, Issue 2 (Summer 2008)
Abstract
Abstract
Background and Objectives
Hemophilia B Leyden is an X chromosome-linked bleeding disorder characterized by an altered developmental expression of blood coagulation factor IX. This form of hemophilia has been found to be associated with a variety of single point mutations encompassing a 40-nucleotide region in factor IX promoter region. Mutations in factor IX gene promoter though relatively rare (about 2% of total) are important because they can give rise to the unique hemophilia B Leyden phenotype.
Materials and Methods
Our objective was to study mutations in exon-1 in 4 3 Iranian hemophilia B patients to recognize possible cases of hemophilia B Leyden. Exon-1 of factor IX gene was amplified by PCR then, conformational sensitive gel electrophoresis (CSGE) was used to distinguish cases having mutations in this region.
Results
Two cases showed band shifts on CSGE. Exon-1 of the patients was directly sequenced. We found two different mutations in exon-1: the A/T mutation at +6 and the A/G mutation at +13.
Conclusions
The prevalence of hemophilia B leyden in B hemophilia patients (4.6%) in our results shows a higher frequency rate in Iran compared to that of other reported countries.
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Key words : Hemophilia B leyden, Factor IX, Promoter region�
S. Babashah, S. Jamali, Dr R. Mahdian, M. Hayat Nosaeid, Dr. M. Karimipoor, Dr. F. Maryami, M. Raeisi, R. Alimohammadi, Dr. S. Zeinali,
Volume 5, Issue 4 (Winter 2009)
Abstract
Abstract
Background and Objectives
Although beta thalassemia is mainly caused by mutations involving single base substitutions and small deletions, there have been reports showing deletions of large regions of beta- globin genes play a similar causing role. The strategy to identify beta thalassemia carriers with known deletions is based on PCR techniques such as Gap PCR. There are however some unknown deletions that can not be detected by the above methods. To overcome this limitation, Real-time PCR and MLPA were developed as two quantitative assays for analysis of beta-globin gene cluster.
Materials and Methods
The subjects were evaluated in a case-control study. Among individuals referred to genetic laboratories of Pasteur Institute of Iran and Kawsar Genetic Research Center, 40 were suspected of having a large deletion in β-globin gene cluster. The including criteria were hematological findings such as low blood indices (MCV <80 fl and MCH <27 pg), normal HbA2 and raised or normal HbF. Genomic DNA was extracted from peripheral blood. A Real-time PCR assay was developed using comparative threshold cycle (Ct) method for analysis of gene copy number. In addition, gene dosage was analyzed using MLPA method.
Results
Real-time PCR results for quantitative analysis of Beta, Delta, G-gamma genes showed the ratio (2-ΔΔCt) of 0.96 ± 0.18 for normal individuals and 0.58 ± 0.04 for carriers of deletions in beta globin gene cluster. MLPA results showed nearly 50% reduction in the height of the peaks corresponding to regions of deletions.
Conclusions
MLPA results confirmed the presence of the same deletions detected by Real-time PCR in all of the carrier individuals. It would be ideal to combine these quantitative assays to confirm corresponding results for accurate diagnosis of known and unknown deletions in beta thalassemia carriers.
Key words : beta-Thalassemia, beta-Globins, Gene Deletion
Dr. H Abolghasemi, Dr. M Aghaiipour, M Nikougoftar, Dr. N Amirizadeh, Dr. M.t Mohammadi, Dr. S Rahmani, F Atashrazm, S Hadjati, P Zarei,
Volume 6, Issue 1 (Spring 2009)
Abstract
Leukoreduction in packed cells filtered by home-made
bedside filters prior and after optimization
Abolghasemi H.1,2(MD), Aghaiipour M.1(MD), Nikougoftar M.1(MS), Amirizade N.1(PhD),
Mohammadi M.T.3( MD),Rahmani S.4(MD), Atashrazm F.3(MS), Hadjati S.3(MS), Zarei P1.(BS)
1Iranian Blood Transfusion Organization, Research Center, Tehran, Iran
2Baghiatollah University of Medical Sciences, Tehran, Iran
3Iran University of Medical Sciences, Tehran, Iran
4Helal Iran Medical Devices Company, Karaj, Iran
Abstract
Background and Objectives
Existence of leukocytes in all blood products causes a wide variety of side effects after transfusion. As a consequence, the use of filter technology for leukoreduction has been widely practiced. In this study, absolute leukocyte count in three types of home-made bedside filtered packed cell units is evaluated.
Materials and Methods
Ninety three packed cell units from blood donors were prepared in Tehran Regional Educational Blood Transfusion Center, stored at 4ºC, and filtered by two types of home-made filters within 1 hour at room temperature. The first type of filters was made prior to 2007 and the second type underwent optimization after 2007. Eight samples were filtered by control group filters with CE certificate. The results were analyzed with SPSS 11.5 and chi-square test.
Results
The mean values of leukocyte count/unit by CD45 and True Count Method were respectively 9×106 and 10×106 in 55 bags filtered by the first type filters and these values were 4.2×106 and 4.8×106 in 30 bags filtered by the second type filters, whereas the mean value of leukocyte count/bag in 8 bags filtered by control filters was 2.3×106.
Conclusions
After optimization of product technology and raw materials, the average number of leukocytes fell within the standard range. Twenty percent of cases contained more than 5×106 leukocytes and 80% less than that (CI95%= 5.7-34.3). Thus, following optimization, the differences in mean rates and SDs in both group two and control filters showed significant reduction and were measured to be within AABB standards.
Dr. M. Shaiegan, S. Mohammadi, Sh. Samiee, Dr. K. Alimoghadam, Dr. Gh. Babaei, A. Ghashghaiee, Sh. Rostami, Dr. F. Kkatami, Dr. A. Azarkeivan, Dr. S. Zolfaghari, Z. Ataiee, Dr. A. Ghavamzadeh,
Volume 6, Issue 1 (Spring 2009)
Abstract
Abstract
Background and Objectives
It is suggested that HA-1 mismatching among hematopoietic stem cell recipients-donors be associated with acute graft-versus-host disease (aGVHD). So the aim of this study was to evaluate HA-1 frequency and examine the correlation between HA-1 disparity and GVHD patients who received transplantation from HLA-A2 identical siblings.
Materials and Methods
Extracted DNA samples were collected from 55 HLA-A2-positive donor-recipient pairs. All the patients received peripheral blood stem cell transplant (PSCT) from HLA-identical siblings. HA-1 was detected by SSP-PCR method. The HA-1 typing was performed using SSP method. Data were analyzed using Chi-square, Man withney and Z test with SPSS 11.5.
Results
Thirty patients showed to be GVHD I-IV and 25 pairs were without any GVHD signs. The frequency rates of HA-1R and HA-1H alleles in patients were 0.55 and 0.45, respectively it showed no significant difference with the frequency rates (0.53 and 0.47) of this allels in donors (p>0.05). HA-1 disparity was detected in 8 out of the 55 donor/recipient pairs (14.5%). aGVHD ( grades I-IV ) was occurred in 6 of patients. Two patients with HA-1 disparity did not show any GVHD signs. X2 test showed there was not any relationship between the incidence of acute graft-versus-host-disease (aGVHD) and HA-1 incompatibility in the patiens.
Conclusions
In spite of higher frequency of HA-1 disparity in GVHD+ group, our data did not reflect any significant association between HA-1 disparity and risk of acute GVHD.
Key words : Polymerase Chain Reaction, Graft- Versus – Host Disease, Minor Histocompatibility Antigens , Hematopoietic Stem Cell Transplantation
Dr. H. Abolghasemi, M. Amani, A. Jabari, R. Ranjbaran, M.h. Mohammadi, Dr. M. Habibi Roudkenar, A. Ali Balazadeh, A. Hashemi Teir, Dr. N. Amirizadeh, Dr. P. Eshghi,
Volume 6, Issue 3 (Autumn 2009)
Abstract
Abstract
Background and Objectives
Fibrin sealant (FS) is a plasma derived product and has hemostatic, sealing and healing properties and is frequently used to reduce blood loss during and after surgery. Blood bank autologous fibrin sealants have no risk of transfusion transmitted diseases. The aim of this study was to obtain of thrombin and fibrinogen from FFP and study their in vitro properties for preparation of FS.
Materials and Methods
Fibrinogen was precipitated by use of protamin sulfate. Fibrinogen concentration was assayed with an enzyme-linked immunosorbent assay and clotting clauss method the effect of temperature on fibrinogen precipitation was also evaluated. Thrombin was prepared by manual method and TPD and its activity was then determined using specific chromogenic substrate spectrophotometric assay. Thrombin stability at different temperature degrees was evaluated and clotting time was measured. Tensile strength and adhesion strength were evaluated with the tensiometry device. Clot lysis time was determined by a clot solubility test in 5M urea.
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Results
Fibrinogen concentration precipitated with protamin sulfate was measured as being 73 ± 8 mg/ml. The recovery of fibrinogen in cryoprecipitate was 93%. Thrombin mixed with fibrinogen had clot time of less than 5 seconds. Tensile strength and adhesion strength of fibrin sealant were 60 ± 8.9 g/cm2 and 55 ± 9 g, respectively. The average activity of the thrombin produced manually was 59.6 ± 6.2. Adding anti-fibrinolttic agent to fibrinogen concentrate has no effect on clotting time and tensile strength but it causes the stability improvement of fibrin clot.
Conclusions
Fibrinogen and thrombin prepared in this experiment have appropriate properties for production of fibrin sealants.
Key words : Fibrin sealant, Fibrinogen, Thrombin, Plasma�
Y. Yahyavi, Dr. H. Teimuri, M. Amani, R. Halabian, M. Edalati, M. Mohammadipoor, P. Hamedi Asl, Dr. N. Amirizadeh, Dr. M. Habibi Roudkenar,
Volume 7, Issue 3 (Autumn 2010)
Abstract
Abstract
Background and Objectives
There is growing evidence indicating that growth factors derived from platelets can be used in wound healing. This study aimed to investigate whether old platelets can be used as the main material for preparation of platelet gel and as substitute for FBS and FCS in cell culture medium.
Materials and Methods
In this exprimental study, platelets were prepared from voluntary blood donors by centrifugation. To prove the hypothesis that the platelet gel and the growth factor derived from expired platelets are able to propagate different cells, platelet derived factors were prepared from both new and expired platelet-rich plasma. The concentration of platelet-derived growth factors was measured by ELISA and cell proliferation was measured by MTT assay.
Results
The results showed the high quality of platelet gel obtained from old platelets. Our results also revealed that old platelets released growth factors similar to those released by new platelets. The growth factors derived from old and new platelets had the same proliferation effects on MSC, CHO, and Fibroblast cell lines.
Conclusions
Old platelets released the same growth factors that new platelets did this showed that old platelets as valuable constituents of blood are cost effective to be used.
Key words : Platelet–Rich Plasma, Platelet–Derived Growth Factor, Cell line
F. Rahmi Nezhad, R. Ali Mohammadi, P. Fuladi, S. Foroughi, M. Feiz Pour, R. Vahidi, M. Hashemi, F. Mola Zadeh, M. Heidari, M. Deilam Salehi, Dr. S. Zeinali,
Volume 7, Issue 4 (Winter 2011)
Abstract
Abstract
Background and Objectives
Beta-thalassemia is one of the most common single-mutation diseases. These mutations cause decreased beta-globin protein synthesis or even deletion. Most of the mutations in beta-globin gene are point mutations but there are some small and big deletions in beta-globin gene cluster. The goal of this research is to detect and study two deletions (Lepore, Asian-Indian) with duplex-PCR.
Materials and Methods
This study was done on 3 Asian-Indian deletion samples and 10 Lepore samples. After detecting two deletions (Lepore and Asian-Indian), we started setting Gap-PCR in these two known deletions. Duplex-PCR with 8 different primers for detecting Lepore and Asian-Indian deletions simultaneously was set at 68ºC annealing temperature these two deletions can be detected easily in one reaction with this new method.
Results
The results show that detecting Lepore and Asian-Indian deletions with multiplex PCR is practical, and the specific bands for each deletion are detectable.
Conclusions
Detecting these two deletions in one reaction (Duplex-PCR) is practical. This method is easy, reliable, and fast it is also economical and saves both time and money.
Key words: Beta-thalassemia, Beta-globins, Gene deletion
Dr. H. Karami, Dr. M. Kosaryan, Dr. H. Abolghasemi, Dr. F. Rashidighader, Dr. K. Vahidshahi, Dr. M. Dabirian, Dr. H. Karami, Dr. M.r. Mahdavi, Dr. E. Yousefi, Dr. R. Alizadeh-Navaei, Dr. A. Tale, S. Shah Mohammadi,
Volume 7, Issue 4 (Winter 2011)
Abstract
Abstract
Background and Objectives
Iron overload especially in heart is one of the most important causes of death among patients with major thalassemia. Chelation therapy is one of the main therapeutic methods in these patients. This study was done to compare the therapeutic effects of deferoxamine (DFO) and deferiprone combination therapy with deferoxamine alone in the patients of Mazandaran, Iran.
Materials and Methods
In this clinical trial, major thalassemia patients with serum ferritin >3000 ng/ml were divided into two groups and matched based on age, sex, serum ferritin levels and cardiac systolic function (LVEF). First group received DFO alone and second group received combination therapy of deferiprone and deferoxamine. The patients were physically examined every month serum ferritin, mean Hb, AST, ALT, and LVEF also started to be measured 6 months prior to the study and subsequently at every visit. Data were analyzed using t-test and χ² test.
Results
There were fifty patients in each group in single and combination therapy groups. Duration of F/U was 28.5 ± 6.2 months. Serum ferritin levels in single and combination therapy groups were 4100 ± 1400 and 4500 ± 1700 ng/ml during 6 months before the study, respectively the levels decreased to 4600 ± 2200 and 3800 ± 1400 ng/ml at the end of treatment, respectively (p<0.05). Only 3 patients had leukopenia and 10 had nausea in combination therapy group.
Conclusions
The study showed that the combination therapy has a better effect on decreasing the serum level ferritin. Moreover, it can improve LVEF.
Keywords: Deferoxamine, Deferiprone, Thalassemia major, Ventricular ejection fraction, Ferritins
A. Khaleghparast, Dr. S. Morovvati, Dr. Z. Noormohammadi,
Volume 8, Issue 2 (Summer 2011)
Abstract
Abstract
Background and Objectives
Two factors known to cause thrombophilia in women with unexplained recurrent spontaneous abortion (RSA) are MTHFR polymorphisms including C677T and A1298C. This study aimed to determine the association between RSA and two polymorphisms of MTHFR in Iranian patients.
Materials and Methods
In this case-control study, 30 patients with the background of two or more consecutive unexplained abortions and 10 women with at least two live births without a miscarriage who referred to Baqiyatallah Hospital and Avicenna Infertility Clinic were analyzed for MTHFR C677T and A1298C polymorphisms by the PCR-RFLP method. Results achieved from estimating the genotype of each polymorphism were analyzed by the SPSS16. The Spearman method was also used to evaluate the correlation between the two polymorphisms.
Results
Data has shown a significant correlation between MTHFR C677T and MTHFR A1298C polymorphisms. Seventeen women (56.6%) with recurrent spontaneous abortions and 5 women (50%) among the controls were heterozygote for MTHFR C677T polymorphism. T allele frequency in the patient group was more than the control group (28.4% for patients and 25% for controls). Frequency of the MTHFR A1298C polymorphism was 63.3% in patients and 50% in controls. For A1298C polymorphism, 43.3% of patients and 20% of controls were heterozygote. Furthermore, 20% of patients and 30% of controls were homozygote for this polymorphism.
Conclusions
The prevalence of MTHFR C677T and MTHFR A1298C polymorphisms were slightly higher in RSA patients compared to controls. These findings failed to support the relationship between thrombophilia polymorphisms and the increasing risk of RSA in evaluated Iranian women.
Sh. Zeighmi Mohammadi, Dr. E. Zeinali, Dr. H. Esmaeili, Dr. A.r. Nikbakht Nasrabadi,
Volume 8, Issue 3 (Autumn 2011)
Abstract
Abstract
Background and Objectives
Exposure to infectious blood and body fluids increase risk of occupationally acquired HIV among nurses. Discrimination in care and treatment of AIDS patient is one of the challenging ethical issues in nursing. The aim of this study was to determine fear of being at risk of acquiring HIV, willingness to care, and discrimination in care and treatment of AIDS patients among nurses.
Materials and Methods
In this descriptive – cross sectional study, 165 nurses of internal and infectious wards from four selected hospitals of Tehran and Shahid Beheshti University of Medical Sciences participated. Data were collected by self-administered questionnaires. The instruments used included "demographic data form", "risk perception scale", "willingness to care for people living with HIV/AIDS"questionnaire, and "discrimination against AIDS" questionnaire. Data were analyzed by using SPSS 14, t-test, and pearson correlation.
Results
Out of 165 nurses, 36.4% had sever fear of being at risk of acquiring HIV, 81.8% were evaluated to be neutral in willingness to care of AIDS patients, and 54.5% agreed with moderately discrimination practice against AIDS patients. There was a significant correlation between fear of being at risk of acquiring HIV with discrimination in care and treatment of AIDS patients (p=0. 003) and willingness to care (p= 0.007).
Conclusions
It seems that education efforts about universal precautions, ethical issues, and patient rights should be made in order to reduce fear of being at risk of acquiring HIV, decrease discrimination in care and treatment of AIDS patients and increase willingness to care.
M. Ashki, Dr. N. Amirizadeh, Dr. M.a. Jalili, Dr. N. Hayati Roudbari, M.h. Mohammadi, M. Amani,
Volume 8, Issue 4 (Winter 2012)
Abstract
Abstract
Background and Objectives
During differentiation of mesenchymal stem cells (MSCs) into various cells, the expression of a variety of genes undergoes some changes in this study we decided to investigate the expression rate of some genes like osteopontin (OPN) and osteocalcin (OCN) during this process in order to find a better and faster way for these cells to be differentiated into osteoblasts .
Material and Methods
In this experimental study, the mononuclear cells of bone marrow were separated and then cultured in DMEM-LG culture media with 10% FBS. During some definite days, the RNA of differentiating cells was extracted. Then, the effective genes in osteogenesis like OPN and OCN were amplified by speciefic primers. The mesenchymal cells were cultured on 3D calcium phosphate scaffolds, and finally the activity rate of the alkaline phosphatase was examined .
Results
This research has demonstrated that in the process of differentiation, the expression of the two genes of OPN and OCN changed orderly with the maximum expression of OPN in the 6th day and the maximum expression of OCN in the 7th and 8th days of differentiation. The osteogenic differentiation of MSCs was not confirmed by the coloration of mineral sediments. The activity rate of alkaline phosphatase revealed the preference of 3D calcium phosphate scaffold to 2D environment in this differentiation.
Conclusion
The calcium phosphate scaffold positively affects the differentiation process. The expression of OPN and OCN genes changes during differentiation and can be used as away to a better and faster differentiation of these cells into osteoblast.
A. Dehghani Fard, N. Saki, M. Ahmadvand, M. Mahmoodinia Maymand, M. Mosahebi Mohammadi, Dr. M. Soleimani,
Volume 8, Issue 4 (Winter 2012)
Abstract
Abstract
Background and Objectives
Mesenchymal Stem Cells have extensive potential to proliferate and differentiate into different cell lineages. Their differentiation capability in vivo and in vitro makes them ideal tools for tissue engineering and regenerative medicine.
Materials and Methods
In the present study more than 100 recent published articles which are about isolation, culture and differentiation of MSCs were reviewed for application of MSC in regenerative medicine and tissue engineering.
Results
Clinical applications of MSCs seem to be in two distinctive lines: bio-scaffold design without immunological responses as well as multipotent stem cell without clinical obstacles.
Conclusions
MSCs due to their capacity of self-renewability, multilineage differentiation and immune modulatory effects are of great therapeutic potential for cell and gene therapy of congenital and degenerative disorders.
M. Mohammadzadeh, R. Halabian, M. Mohammadipoor, A.a. Kiani, Dr. A. Gharehbaghian, Dr. N. Amirizadeh, Dr. M. Habibi Roudkenar,
Volume 8, Issue 4 (Winter 2012)
Abstract
Abstract
Background and Objectives
Poor viability of Mesenchymal Stem Cells (MSCs) following transplantation is one of the major challenges in their therapeutic application. Manipulation of MSCs by the genetic engineering method is one of the strategies used to protect the cells against cytotoxic microenvironment. However, maintaining multi differentiation capacity of MSCs following manipulation is important. We investigated if the manipulation of MSCs with NRF2 affects the multi differentiation capacity.
Materials and Methods
MSCs were isolated from bone marrow. NRF2 was isolated and TOPO cloned into the pENTR vector. The recombinant vector was transferred into pAD/CMV/V5-DEST vector by gateway technology. Recombinant adenovirus was produced in AD293 cells, followed by being infected into MSCs. Expression of NRF2 was verified by RT-PCR. The NRF2 engineered MSCs were exposed to stress conditions followed by the evaluation of the cells viability and apoptosis. Finally, NRF2 expressing MSCs differentiation into osteoblast and adipocyte lineages was studied.
Results
NRF2 was successfully expressed in MSCs. NRF2- MSCs differentiation into osteoblast and adipocyte lineages indicating overexpression of NRF2 does not affect the differentiation property of MSCs.
Conclusions
Expression of NRF2, a well known cytoprotective factor, by using adenovirus expression system does not intervene in the differentiation capacity of MSCs. NRF2-MSCs might be applicable for stem cell-based cell therapy in future.
M. Paryan, Dr. M. Fourozandeh Moghadam, S. Mohammadi-Yeganeh,
Volume 9, Issue 1 (Spring 2012)
Abstract
Abstract
Background and Objectives
Hepatitis C is the most common viral disease among intravenous drug users transmitted mainly through blood transfusion. In this study, we used an isothermal nucleic acid sequence-based amplification (NASBA) in combination with a molecular beacon probe-based real-time assay for detection of HCV.
Materials and Methods
In this experimental study, the conserved 5’NCR region with the length of 241 bp was used for probe and primers design. In comparison with the standard, RNA virus detection sensitivity of the method was 100 percent for up to 500 copies/ ml.
Results
No serological positive sample was detected with this assay the clinical sensitivity of reaction was 96.6%. NCBI Nucleotide Blast showed 100% analytical specificity and no cross was observed with viruses or human genome. Investigation of 10 serological negative samples showed the clinical specificity to be 100%.
Conclusions
In summary, due to its isothermal nature, its speed, and its use for RNA specific amplification, NASBA real-time assay will have broad applications for the rapid detection of HCV in plasma samples.
.m Tajvidi, Sh. Zeighmi Mohammadi,
Volume 9, Issue 1 (Spring 2012)
Abstract
Abstract
Background and Objectives
Beta-thalassemia major is the most prevalent genetic blood disorder in Iran. Several studies showed that chronic diseases have a negative impact on self-esteem, social and interpersonal relationships, and psychological health. The purpose of this study was to determine the level of loneliness, hopelessness, and self-esteem in thalassemic adolescents.
Materials and Methods
In this descriptive cross sectional study, 100 thalassemic patients participated through 2010. They were selected by convenience sampling from the Center for Special Diseases. The data were collected by self report. The instruments used in this study included the sampling and demographic data form, Beck Hopelessness Scale, Coopersmith Self-esteem Scale, and Loneliness Scale. Data were analyzed by SPSS 14 software and using descriptive statistics, t- test, ANOVA, and coefficient Pearson correlation.
Results
The findings indicated that the mean score of hopelessness was 6.12 ± 3.83, loneliness41.66 ± 5.12, and self-esteem 100.86 ± 14.04 59% of thalassemic patients had low hopelessness, 51% moderate loneliness, and 55% moderate self esteem. There was a statistical significant correlation between the score of hopelessness and self-esteem (p=0.027), but there were not any statistical significant correlations between hopelessness and loneliness and between self-esteem and loneliness.
Conclusions
The results showed that thalassemia had a negative influence on the psychosocial aspects of adolescents’ life. Knowledge of the mechanisms and strategies to prevent psychosocial disorders and promote mental health of thalassemic patients is necessary. Nurses by assessing psychosocial needs, providing necessary psychosocial support, and making appropriate intervention can help promote mental health of thalassemic patients.