THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)
Year:2006 | Volume:3 | Issue:2|Papers Count:15

Background and Objectives: RhD antigen has an important role in causing hemolytic diseases in newborns; in testing fetal RhD status by an invasive method such as amniocentesis, it would also harm the fetus and mother. This study tried to detect fetal RhD genotype using free fetal DNA in maternal serum by a noninvasive method using hemi-nested PCR. Materials and Methods: Blood samples were collected from 45 RhD- negative pregnant women whose spouses were Rh-positive; the women's serum free DNAs were isolated by Phenol-Chloroform method. The exon 10 of the RhD genes was amplified in two rounds of PCR using special primers. For confirmation of negative results obtained in the second PCR, the fragment of the RhCE gene was amplified using special primers. Results: Follow up and evaluation of the PCR results and relevant serological analysis of cord blood after delivery revealed that in 37 out of 41 cases the RhD had been determined appropriately (95.45%). There were one false-negative and three false-positive cases. Conclusions: This study showed that maternal serum is very suitable for prenatal diagnosis of fetal RhD using noninvasive methods with minimum risk for neonates. In this regard, modern molecular methods with high sensitivity such as hemi-nested PCR and elimination of interfering factors could be applicable in clinical approaches.

Background and Objectives: Human plasma is a valuable and unique material containing vital proteins such as albumin, immunoglobulin, and coagulation factors which have pharmaceutical applications. The most current process for purification of immunoglobulin is the Cohn fractionation procedure using ethanol. The basic material for production of immunoglobulin in this procedure is fraction II. In the present study, Cohn fractionation was modified appropriately in order to achieve fraction II with a quality suitable for production of intravenous immunoglobulin. Materials and Methods: Various plasma proteins were precipitated using varying concentrations of ethanol under controlled physiochemical conditions such as temperature, pH and ionic strength. The fractionation products as well as the resulting fraction II were tested and analyzed. Results: The results demonstrate that the modified procedure used can result in a pure fraction II with high yield. The procedure was also suitable for all other plasma proteins. The purified fraction II contained acceptable PKA levels according to specifications of pharmacopoeia. The protein structure was also within normal limits. The repeatability of the modified procedure reported was acceptable.Conclusions: The fraction II obtained in the modified Cohn procedure was a suitable intermediate product to be used in production of intravenous immunoglobulin. The PKA levels as well as the protein aggregation were minimal, and the quality of fraction II was standard. The results showed that the present modified Cohn procedure can be easily scaled up to industrial and semi-industrial levels. The resulting fraction II can be used as an intermediate in production of IVIg.

Background and Objectives: Transfusion of blood products is associated with a number of changes in metabolic parameters. The aim of this study was to assess the effects of transfusing packed red blood cells on arterial blood gas parameters in the patients undergoing general anesthesia for elective surgery.Materials and Methods: In a clinical trial, each one of the 40 adult cases undergoing general anesthesia for elective surgeries received a unit of packed cell with a volume of 250 to 350 ml. The arterial samples were taken from these recipients exactly both before and after the transfusion. P value less than 0.05 were considered significant; data entry and analysis were done using SPSS software version 11.5. The results were demonstrated as mean ± standard deviation. Chi square and paired sample T tests were used for data analysis. Results: There was no statistically significant difference between the two samples regarding arterial values of pH, Base Excess, PaO2, and PaCO2 before and after transfusion of one unit of packed red cell. There was, however, a significant decline in the arterial amount of bicarbonate and Buffer Base after the transfusion. Conclusions: Packed cell transfusion as much as one unit or so in those patients undergoing general anesthesia could not influence the blood gas parameters in such a notable way. However, it seems that there is no reason to administer any routine dose of intravenous bicarbonate after such a transfusion, unless indicated otherwise by arterial blood gas documentation.

Background and Objectives: Bone marrow of adults contains a subtype of progenitor cells that has the capacity to differentiate into mature endothelial cells and has therefore been termed as endothelial progenitor cells (EPCs). Clinical studies employing EPCs for revascularization of ischemicorgans have just started to be conducted. In the present research, endothelial progenitor cells were isolated from peripheral blood and transfected by lipofection method. Materials and Methods: All mononuclear cells were isolated from peripheral blood by vivo expansion method and cultured in fibronectin coated dishes and basal EC medium. In day 7, the cells were tripsinized and examined by immunocytochemical and molecular evaluation methods; then, electroporation and lipofection methods were used for transfection of EPCs. Results: Observations showed isolated cells were proliferated on fibronectin coated surfaces. The cells were able to incorporate Acl-Dil and express CD31, CD34, KDR, VECAM-1, Tie-1 genes, CD31, CD34, Lectin and KDR antigens. The cells were also transfected by lipofection method. Conclusions: It can be concluded that hEPCs express specific endothelial genes in day 7 and are transfected by lipofection method with low efficiency.

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 eliminate blood replacement and it has been considered as a priority in the strategic planning 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.

Background and Objectives: Umbilical cord blood (CB) has been identified both as a rich source for hematopoietic stem cells (HSCs) and as an alternative to bone marrow transplantation. To accelerate the pace of engraftment in cord blood (CB) transplantation, HSCs should be ex vivo expanded. So the aim of this study was to evaluate the effects of macrophage inflammatory protein-1 alpha (MIP1- α) on ex vivo expansion of HSCs and its role in prevention of HSCs differentiation in different conditions.Materials and Methods: Mononuclear cells (MNCs) were seperated from cord blood and cultured in RPMI 1640 with 10% fetal calf serum (FCS) or 10% cord blood plasma (CBP) or serum free media (SF). Culture media contained 50 ng/ml of interlukin 6 (IL-6), interlukin 3 (IL-3), thrombopoietin (TPO), stem cell factor (SCF), and flt3-ligand. MNCs were cultured for 2 weeks in the presence or absence of MIP1-α in CO2 incubator; expansion potential was then studied by MNCs counting and flowcytometry for CD34+/CD38- cells in 7 and 14 days.Results: In all MIP1-α contained media, more increase in expansion of CD34+/CD38- and CD34+ cells was observed.Conclusions: These results suggest that the effect of MIP1- α depends on type of medium and the cytokines present in the culture; it also shows that addition of MIP1-α is useful for clinically ex vivo expansion of cord blood-derived progenitor cells.

Background and Objectives: Blood transfusion has a critical role in clinical practice, but there has always been a possibility for transmission of infections. A lot of research has been conducted recently to find the cause of these infections. In 1997, TTV was first found by Nishizawa. Our aim was to estimate the prevalence rate of TTV in healthy blood donors and recipients of blood components in Tehran and prepare PCR kits to detect this virus.Materials and Methods: In this research, we studied the prevalence rate of TTV in 250 thalassemic, hemophilic and dialysis patients and 250 blood donors. After extraction of DNA and amplification by seminested polymerase chain reaction, the bonds of DNA were observed in electrophoresis with 2% gel agarose. Results: 250 patients were studied 66.9% of whom were positive and 33.1% negative for TTV. TTVPCR results were also studied in 250 blood donors 41% of whom were positive and 59% negative. There was a significant difference (p=0.0001) between patients and the control group.Conclusions: TTV has a high prevalence in recipients and blood donors. Blood transfusion probably is not the only way for transmission of TTV, and other ways such as oral-fecal route can also play a role for transmission of TTV.

Background and Objectives: White blood cells (WBCs) in blood products are the major stimulus for a number of detrimental biological reactions including febrile nonhemolytic transfusion reactions, alloimmunization against HLA antigens, and cytomegalovirus transmission. In this study, our objective was to study the effect of storage time on the filtration of platelet concentrates (PCs). The total number of white blood cells as well as the distribution of WBC subsets in PC units filtered before and after storage were compared.Materials and Methods: In this experimental study, platelet-rich plasma-derived PCs (5 pooled units) were filtered both fresh and after 5 days of storage; then, the total number of WBCs was calculated by flow cytometry and cell counting. WBC subsets were analyzed by flow cytometry with three-color fluorescence. In this study, the third generation bags and filters were used.Results: Before filtration, the total number of WBC was significantly higher in fresh units compared with stored units, whereas in postfiltration samples the number of white blood cells was significantly lower in the fresh compared with the stored units. Although the absolute number of WBCs significantly reduced, filtration induced significant changes in proportion in subsets in both fresh and stored units; the percentage of B cells and monocytes increased after filtration.Conclusions: Both pre- and post-storage WBC filtration affects the proportions of WBCs in the final product but pre-storage WBC filtration of platelet concentrates is superior than post-storage WBC filtration.

Background and Objectives: High risk and unaware donors embarking on blood donation endanger blood safety. Thus the importance of donors training for providing safe blood is one of the main objectives of IBTO. In this study, we evaluated the awareness of blood donors about safe blood donation among Tabriz blood donors in 1384.Materials and Methods: This cross-sectional study was conducted on 1200 literate blood donors who had studied the screening educational materials designed to raise their awareness about safe blood donation in Tabriz. The blood donors were selected using random sampling and a 3-part questionnaire was distributed among them. Part one contained questions about demographic characteristics, part two aimed to evaluate the method the materials were presented, and part three intended to evaluate awareness of donors about safe blood donation. Statistical analysis was performed using SPSS (version 11.5) and chi-square test.Results: 97% of blood donors answered the questions designed to evaluate the method the educational materials were presented; the answers showed their satisfaction. There was a correlation between educational level of donors and the degree of using educational pamphlets, the degree of their evaluation about the simplicity of materials and their length, and the degree of learning new information about blood safety (p<0.05). 68.8% of blood donors knew that routine screening tests cannot detect a recent infection (window period) and 66.8% were aware of confidential self-exclusion. There was a correlation between the number of donations and awareness of window period and confidential self-exclusion (p<0.05). The most frequent questions receiving incorrect answers and being left unanswered were about window period of transfusion - transmitted diseases and confidential self-exclusion that came out to form 31.2% and 33.2% of the total number of questions respectively. In addition, 51.7% of blood donors thought that the most important factor to ensure safe blood is the laboratory testing rather than consultation with physician.Conclusion: Since the educational level of blood donors is an important variable in evaluation of the method the educational materials are presented, it should be considered in providing screening educational materials. The results of the study about respondents' awareness of window period and confidential self-exclusion show that these concepts need to be revised in educational pamphlets. Moreover, the sincere collaboration with consulting physician as an important factor to ensure safe blood should be further emphasized.

Background and Objectives: Increasing rate of infection through blood-transmitted agents, especially viruses such as HIV, HCV and HBV and long duration of their window period were the most important reasons for implementing the confidential self-exclusion system. This system is used to help blood donors exposed to contamination be able to exclude their blood units confidentially by informing blood centers. This research was conducted in Qom Blood Center in order to study the efficiency of the confidential self-exclusion system to improve blood safety.Materials and Methods: The comprehensive data bank of Iranian Blood Transfusion Organization was used to collect information about 12935 blood donors during the period the confidential self-exclusion system applied from 01.09.82 to 31.03.83 (according to Iranian calendar) in this cross-sectional study. Data were analyzed by applying SPSS software and Chi-square statistical test.Results: Out of 12935 donors, 204 (1.57%) had embarked on excluding their blood. Out of this number, 22 (10.78%) were ELISA positive and 182 (89.22%) ELISA negative. Out of 22 blood donors whose screening test results were positive, 11(50%) was HBs-Ag positive and the other 11 (50%) HCV-Ab positive; none of them (22 blood donors) was positive in HIV-Ab and RPR test. Using Chi-square tests to analyze data, it came out to be evident that there was no significant statistical correlation considering marriage status, age and sex between the results of screening tests (ELISA) for blood donors who had excluded their blood donations and those who had not (p>0.05). Conclusions: Given the results of carrying out confidential self-deferral system in Qom and some other blood transfusion centers across the country and increasing rate of infection around the world especially the third world countries, implementation of this plan in order to improve blood safety is imperative. Holding relevant educational workshops to raise public awareness and personal knowledge would further increase blood safety.

Background and Objectives: Iron is an essential element which plays its role in metabolic pathway in the body and it is required by every human cell. Main site of iron lies in the structure of hemoglobin in red blood cell. Blood donation has a marked influence on body iron stores in female blood donors leading to anemic blood donors unable to donate; moreover, the anemia may trigger a costly medical evaluation. This study was conducted to evaluate the frequency of iron deficiency anemia in first time and frequent female blood donors referring to Tehran Blood Transfusion Center.Materials and Methods: In this study (historical cohort), 69 first time female blood donors as the control group and 75 frequent female blood donors as the case group were randomly selected. Hemoglobin in the two groups of blood donors was tested by Hb strip and all of them were considered eligible for donation. Blood samples of blood donors were transferred to the laboratory for the evaluation of biochemical parameters for iron status in the body and cell blood counting. These data were compared between the two groups by using SPSS statistical data program. Results: The age range in the two groups was significantly different (p<0.001). Therefore, female blood donors were divided in two groups according to age: above and below 45 years of age. There was a significant decrease in Hb level in frequent blood donors when compared with the Hb level in first time female blood donors at age range below 45 years (p<0.003). According to the results, frequency of anemia was 10.7% in frequent female blood donors and 6.6% in first time female blood donors. Serum transferrin and TIBC levels in frequent female blood donors were significantly higher than the first time blood donors (p<0.003, p<0.006). We found significant decrease in ferritin level at age range below 45 years among frequent female blood donors when compared with ferritin level in first time female donors (p<0.04).Conclusions: The results show that blood donation has a marked influence in decreasing ferritin and increasing transferrin levels in frequent female blood donors which indicates complete depletion of iron in their bone marrow. Ferritin and transferrin measurement is an important factor for diagnosis of iron deficiency and should be included in the routine assessment of female blood donors. We recommended that the frequency of phlebotomy be adjusted according to levels of serum ferritin and serum transferrin.