The Scientific Journal of Iranian Blood Transfusion
Year:2012 | Volume:8 | Issue:4 (33)|Papers Count:13

Background and Objectives: Activated platelets release microparticles (MPs) in vivo or in the platelet concentrate (PC) product in response to some stimulators. This study compared the two different storage media of plasma and Composol for PC regarding the extent of MPs formation as a new index for comparing platelet stability.Materials and Methods: In this experimental study, 30 PC units were prepared. The platelets were divided into two equal portions. Plasma was replaced with the additive solution of Composol in one of the portions. Sampling was carried out at the days 2, 4 and 7 after storage. Afterwards, the MPs were separated and their concentrations were collected using Bradford method. T-tests was used to compare the results of this experiment.Results: The results showed that the amounts of MPs increased during the storage. In each media of plasma or Composol, the concentration of MPs showed a significant difference between the days 4 and 7 of the storage. Besides, the final concentration of MPs did not show a significant difference between the two media at the day 4 of the storage whereas this difference was significant at the day 7 of storage (p<0.05).Conclusions: There were no significant differences in the quantity of MPs in PCs stored in plasma or Composol up to the day 4 of the storage. This difference became significant after 7 days of the storage, in which the generation of MPs showed significant increase in plasma than Composol. These observations revealed an advantage for Composol as a PC storage medium.

Background and Objectives: FLT3 mutations are associated with poor outcome in acute myeloblastic leukemia (AML) patients. Only limited information is available about effects of FLT3 mutation on Acute Promyelocytic Leukemia (APL). We investigated the prevalence and impact of FLT3 mutations on the clinical characteristics and the response to treatment in APL patients treated with arsenic trioxide (As2O3).Materials and Methods: Blood samples were collected from 115 untreated APL patients and genomic DNA was extracted by the salting-out method. FLT3-ITD and FLT3-D835 mutations were investigated by PCR-RFLP. Mann-Whitney U test and Chi-square were used for data analysis.Results: FLT3-ITD and FLT3-D835 mutations were detected in 16 (14%) and 13(11%) of the patients, respectively. Both mutations were identified in two patients, so overall frequency of FLT3 mutations was estimated to be 23.5%. Patients positive for FLT3–ITD mutation had a higher rate of white cell counts (p=0.005) and more frequent bcr3 type of PML/RARA fusion (p=0.04). We have not found any significant association between FLT3-D835 mutation and the clinical characteristics of patients. Between the group with FLT3 Mutations and the group without, there was no significant difference in response to therapy.Conclusions: Complete remission induction with As2O3 may be independent of FLT3 mutation status, so As2O3 may be the first choice of APL especially in patients with FLT3 mutations. However, further studies on a large group of patients are necessary to confirm our findings.

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.

Background and Objectives: For reducing bacterial contamination of platelets in the medium, FDA has approved the Bact/Alert for screening the platelet units. This study attempts to compare the Bact/Alert system and the manual culture method as far as the length of time in hours of detection is concerned.Materials and Methods: In this interventional and diagnostic study, 15 platelet units were selected randomly among 1332 units and inoculated with 10 CFU/ml of various bacteria including Streptococci, Serratia marcescens, Enterobacter cloacae, Corynebacterium diphteroid, Staphylococcus aureus and Staphylococcus epidermidis which normally contaminate platelet units. These units together with other platelet units in a blind way were tested by both Bact/Alert system and the manual method.Results: Regarding the short expiration time of platelet units, if the length of time in hours in detection is considered as a basis for comparison, the Bact/Alert system is significantly superior to the manual method. The medium length of time in hours for detecting the aerobic bacteria by Bact/Alert system is 31±8 hours, compared with the manual method which is 61±11 hours. This shows that Bact/Alert system is nearly 2 times faster than the manual method.Conclusions: Bact/Alert culture system compared with the manual method is more rapid and accurate for detection of bacterial contamination thereby improving platelet safety. Regarding serious effects of these contaminations on platelet recipients, it is also necessary to try to reduce them by using GMP.

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.

Background and Objectives: Some of the dental procedures can cause bleeding. Bleeding control can be affected in some patients due to systemic disease or chronic anticoagulant therapy, so they may be at increased risk for bleeding events or even death following invasive dental procedures. This study was designed to evaluate the knowledge of general dentists in Qazvin city regarding coagulation tests performed in bleeding disorders during 2010-2011.Materials and Methods: A questionnaire (including 23 questions) was designed with the aid of specialists in the field of oral medicine and hematology. This questionnaire was distributed among 124 general practitioners. Data were analyzed with SPSS version 15 and T-test, one-way ANOVA and Tukey.Results: The mean score for dentists knowledge was 8.64±1.20. There was no significant difference in the mean knowledge scores among male and female dentists. Tukey test showed a significant difference in the mean knowledge level among 31-40 year old and over forty year old dentists (p<0.04).Conclusions: This study showed that knowledge of the dentists regarding bleeding disorders is not at desirable level which requires planning for continuing education courses.

Background and Objectives: Cardiovascular disease (CVD) is a common cause of morbidity and mortality. The relationship between ABO blood groups and main risk factors of CVD is unknown. So this study was designed to investigate whether there is an association between ABO blood groups and cardiovascular risk factors in healthy population.Materials and Methods: In this cross-sectional study, risk factors screening for CVD on 2920 healthy individuals of Golestan province in 2005 were estimated by a questionnaire that aimed to extract information about age, sex, physical activity, smoking, blood group type, weight, height, blood pressure and family history of CVD. Data were analyzed with SPSS13 and by using Chi Square and ANOVA tests.Results: Out of the total number of 2920, 57.4% were male, 70% inactive, 14% smoker, 25% hypertensive, 23% obese, and 21% had family history of CVD with the mean age of 41.52±12.317. Blood groups O (32.9%), A (30.1%), B (23.3%) and AB (13.7%) were the most frequent ones, respectively. Amongst cardiac risk factors, it was only the frequency of family history of CVD that varies across different blood groups, and individuals with A blood group reported to have a more frequent family history of CVD as compared with other blood groups.Conclusions: These findings illustrate amongst cardiovascular risk factors only family history of CVD as having a significant correlation with ABO.

Background and Objectives: Factor XIII is the last enzyme in the clotting cascade. The gene of A chain is located on chromosome 6. Deficiency of factor XIII in autosomal recessive conditions occurs at a frequency of 1 in 2 million general population. The aim of this study was to detect the mutations of subunit A in both patients and carriers.Materials and Methods: In this study we have investigated the molecular basis of inherited FXIII deficiency among patients from 21 unrelated Iranian families. Mutation were detected by amplifying each exon. Those exons exhibiting the presence of hetero duplex formation sensitive gel electrophoresis, were selected for direct sequencing. After sequencing, detected mutation was carried out by restriction fragment length polymorphism (RFLP).Results: All patients having entered the study had mutations. Twelve patients had homologues substitution of TGG®CGG in exon 4, 1 insertion mutation occurring in exon 7 triple G, 2 patients demonstrated mutation exon 9 ATG®AAG, 3 patients had substitution of CGG®CAG in exon 10, and 3 patients showed a homologue subsituation mutation in exon 15 GCC®GTC.Conclusions: Our findings suggest that the activity of enzyme is highly dependent on the core domain. Changes in charge, amino acid tail and conformation lead to decreased enzyme activity. Also tetrameric structure is calcium related. It seems that changes of amino acid sequence convert enzyme stability.

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.