THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)
Year:2005 | Volume:2 | Issue:4|Papers Count:13

Background and Objectives Rheumatoid arthritis (RA) is a severe chronic inflammatory disease that can not be easily rapidly treated. It can cause joint destruction and disability. Generally some laboratory tests, such as Rheumatoid Factor (RF), Erythrocyte Sedimentation Rate (ESR) or C-Reactive Protein (CRP) can be used for diagnosis and monitoring of the rheumatoid arthritis. However, they are not always ideal. In inflammatory diseases such as rheumatoid arthritis, the level of IgA will increase in serum; the surplus IgA can then react with some of the serum proteins such as Alpha-1-antirypsin (α1AT) to form a non-immune complex. The complex IgA – α1AT is formed by disulfide bonding between an active thiol groups available on the both proteins. Some reports suggested that this complex is a good marker for RA disease without false results, while RF test has some false negative results. Materials and Methods In this study the level of IgA-α1AT complex in thirty seven RA patients and in forty four normal subjects by ELISA methods using anti- α1-antirypsin monoclonal and anti-IgA polyclonal HRP conjugated antibodies was evaluated. Routine laboratory tests of RF, CRP and ESR in patients and controls were also investigated. Results Our results showed that IgA- α1AT complex level in patients (43.7 ± 15.4) is significantly higher than controls (21.5 ± 8.3) (p<0.05). There were significant differences between the sera complex levels in patients and controls. The RF results revealed eleven false negatives (30%) while the level of complex had only one false result (3%). Conclusions Instead of RF, a rapid and sensitive ELISA test for IgA- α1AT complex level in RA patients is strongly recommended.  

Background and Objectives The embryonic yolk sac cells have two unique characteristics: high proliferative capacity and lack of MHC associated antigen. According to the positive effects of co-culturing system on the differentiation of stem cells, in this study we evaluate the effect of M2-10B4 stromal cell line and erythropoietin (EPO) on the differentiation of embryonic yolk sac cells to erythroid cells. Materials and Methods The yolk sacs were dissected from 10-day mice and their cells were separated using syring needles and enzyme digestions (0.1% collagenase/20 fetal calf serum (FBS) or 0.25% trypsin – 0.02% EDTA at 37°C). The M2-10B4 stromal cells were cultured in the DMEM medium and mitotically inactivated using mitomycin C. These cells were co-cultured with yolk stem cells in the medium containing stem cell factor (50 ng/ml); EPO (1U/ml) and the colony assay of these cells (5´104 cells/ml) have been done in the presence of interleukin-3 (40 ng/ml), stem cell factor (50 ng/ml) and EPO (1U/ml) in semisolid medium. After 7 days, benzidine staining on the colonies was carried out and positive benzidine colonies were considered as erythroid colonies. Results The colony assay showed that in the presence of EPO the growth of erythroid colonies were better than the other group and they had more benzidine colonies and cells. Conclusions By using the stromal feeder layer such as bone marrow stromal cells and erythropoietin the differentiation and proliferation of YSC was improved; however, further studies are needed.  

Background and Objectives During the infancy period, exchange transfusion is the main and effective method of cure for hyperdilirubineamia in hemolytic neonates. One of the risks involved in these patients is CMV infection. Since CMV can inflict and kill children with immunodeficiency, we decided to evaluate blood transmitted CMV infection in infants for the first time in Iran. Materials and Methods ln this prospective study, the levels of lgG and IgM antibody against CMV in infants blood sera before and after exchange transfusion, sera of their mothers, and the plasma of donors were evaluated by ELISA  method and conventional kits. The data were then analyzed by nonparametric and mac nemar test.  Results The level of specific IgM antibody of CMV was 14% in the serum of infants before exchange transfusion  and  48% two months after exchange which came out to be significant (p<0.0001). This indicates transmittability of CMV infection through blood transfusion since the amount of primary infection of CMV in mothers (IgM) was only 1%. Conclusions According to the high level of CMV infection in infants (14%) compared to mothers (1%), it can be concluded that some mothers have latent CMV infection. On the other hand, the high level of CMV infection in infants (48%) 2 months after transfusion is cause of post-transfusion CMV infection. Analysis of the data obtained from dependent variables such as volume of transfused blood, number of exchange transfusion attempts, weight, age, sex and blood group analyzed by the nonparametric method indicated that the volume of transfused blood to infants has a direct correlation with the rate of transmission of infection to them but other variables do not have any correlation.    

Background and Objectives Hemophilia B is a genetic disorder due to deficiency or complete absence of factor IX coagulation factor. Treatment of choice for these patients is use of factor IX concentrates. Therefore, purification of plasma proteins and separation of factor IX have been major objectives for scientists involved in this field. In this respect, purification procedure using ion exchange chromatography is widely used, but in the past decade affinity chromatography was also introduced. The objective of the present study has been to apply both techniques for the purification of factor IX and compare the quality and yield of the product. Materials and Methods For the purification procedure, chromatography columns (XK-16), containing DEAE sepharose and Heparin sepharose were used. Factor IX coagulation activity was measured using a one-stage coagulation assay and factor IX antigen was quantified using ELISA technique.  Results The specific activity and relative increase in purity of factor IX was calculated and it was demonstrated that specific activity improved from 3.1 IU/mg using DEAE ion exchange to 29 IU/mg when affinity chromatography was added and purity was increased from 155 to 1450 respectively. Conclusions The present study demonstrates that addition of an affinity chromatography step using heparin sepharose is a major improvement in the purification of factor IX, where both specific activity and purity are increased considerably.  

Background and Objectives HTLV is a member of Retroviridae family whose infection can lead to leukemia. Since in most regions of Iran (like Bushehr) donated blood units are not tested for HTLV infection, multiple transfusion patients are at high risk. In this study, we evaluated serological prevalence of the infection among Busherian patients. Materials and Methods This was a descriptive study in which samples of all Busherian multi-transfused patients were taken based on dispersion of thalassemic and hemodialysis hospital wards. Blood samples for detection of HTLV I/II infection markers were tested by ELISA method and for confirmation of positive ELISA results western blot technique was used. Data were analyzed by using SPSS software and Chi-square.  Results Out of 642 Busherian multi-transfused patients including 455 (70.78%) thalassemic, 86(13.3%) hemophilic and 101 (15.7%) hemodialysis patients, 41 all thalassemic were ELISA positive out of whom 22(53.7%) were male and 19(46.3%) female. Infection in 14 cases was confirmed by western blot technique (p=0.77). Statistical analysis did not show any significant correlation between western blot results and gender (p=0.56). Neither was there any significant difference between western blot test results across different age ranges (p=0.67). The significant point in this research was the detection of a thalassemic patient with dual infection of HTLV type I and II. Conclusions Detection of HTLV infection among multi-transfused population indicated that such an infection may exist among blood donors of Bushehr.  

Background and Objectives After hepatitis and AIDS, malaria is the most prevalent transfusion outcome in endemic areas. Presence of asymptomatic carriers of malaria parasites in the endemic areas can be a source of infection in transmission of malaria by blood transfusion. Prevention of malaria caused by blood transfusion depends on screening blood donors and deleting infected blood samples. To screen blood samples, parasitological, serologic and molecular methods have been applied. Materials and Methods In this study 120 blood donors in Iranshahr in Sistan-Baloochestan province were tested with different methods of thick and thin blood films, Immuno-Fluorescent Antibody Test (IFAT), and Polymerase Chain Reaction (PCR).  Results The result of all thick and thin blood films was negative. IFAT by using P.vivax antigen and P.falciparum antigen for 38 and 6 donors respectively showed a titre of antibody equal to± 1/20-1/320 (17 of the former group and 4 of the latter had a history of malaria infection). The PCR assay using silica for DNA extraction and using P.falciparum specified primers with sensitivity rate equal to 2-3 parasites per microlitre of blood was negative for all subjects under study. Conclusions This study showed, although microscopic examination of blood smears was inexpensive and simple, but it is labor-intensive and time-consuming that makes it insensitive for detection of low-level parasitemia in asymptomatic donors and for screening a large number of specimens. IFAT would not always show the real existence of parasites and in spite of simplicity and sensitivity because of its disability to be automated is not suitable for screening a large number of specimens. On the other hand, IFAT in individuals with malaria history and absence of parasites in their blood may be positive for a long period. It was approved that molecular methods such as PCR were more sensitive and more specific than conventional microscopic examination and their great advantage was the ability to detect the infection with low-level parasitemia that may have been distinguished by blood films examination. In the present study, probably because of low number of specimen or limited study duration with PCR method, or probably since parasitemia exiting in the subjects under study was less than 2-3 parasites per microlitre of blood, we were not able to detect positive cases.    

Background and Objectives Patients with major b-thalassemia have been exposed to a wide range of blood-borne viruses among which hepatitis B and C viruses have a considerable importance. In this study, seroprevalence and risk factor of HBV, HCV and HIV infections among thalassemic patients of East Azerbaijan were evaluated. Materials and Methods Eighty four thalassemic patients who were regularly transfused at Children Hospital (Tabriz) enrolled in this observational study. Some data were collected through questionnaires and patients’ medical history; then, patients were evaluated for the presence of HCVAb, HBsAg, HIVAb, HBsAb, HBcAb and ferritin by ELISA; moreover, HCV positive sera were confirmed by Recombinant Immunoblot Assay (RIBA-3.0).  Results None of the eighty-four thalassemic patients were HIV and HBV positive; however, 6 (7.1%) were HCV positive. Distribution of sex and the mean ferritin did not differ from HCV-positive to HCV-negative patients, but the age average was significantly higher in HCV-positive patients (p<0.001). Moreover, HCV-positive patients had a significantly longer history of transfusion compared with HCV-negative patients (p<0.001). The prevalence of HCV in those patients who had received their first blood transfusion before the implementation of compulsory testing for HCV in 1995 was significantly lower. In other words, no case became infected after the initiation of donor screening test for HCV. Conclusions The prevalence of HCV in thalassemic patients of East Azarbaijan was lower compared to other provinces of Iran and all positive cases in this study went back to the time period preceeding 1995 that is before the implementation of compulsory HCV screening tests.

Background and Objectives Setting research priorities in the cycle of research management is critical. The limitation in human and financial resources and policy changes are the most significant reasons necessitating research priorities to be set. Research prioritization can materialize and be effective at different levels ranging from macro and national to educational and research levels. To this end, IBTO Research and Education Deputy by this study have embarked on a serious measure in organizing and orientating investigations in IBTO. Materials and Methods First the necessity of the implementation of the project was elaborated in the Research Council and priorities were set. Then, different procedures were conducted based on the guidelines of COHRED (Council on Health Research for Development) and by use of priority-setting instruments applied in research institutes. At the end, the results were reviewed by the Research Council so that the final priorities were approved of. Results In the present study, out of the whole number of forms distributed for priority-setting, blood centers, headquarter managers and consultants, and the faculty members had respectively a share of 64.28%, 33.33%, and 25.92% in responses. At the process of title collection, more responses were received as compared with the priority setting process. Finally, 99 research titles in 16 domains were approved of as final priorities by the IBTO Research Center. Conclusions Priority-setting was conducted through the method recommended by COHRED for the first time in IBTO. In spite of the participation of the out-of-organization beneficiary, research centers, and scientific associations, the highest rate of participation goes to intra-organizational groups. Approved priorities can be implemented by a call for research, the creation of an evaluation system for recommended projects, and survey of approved projects. Thus, the most use can be made of financial and human resources for priority-setting.  

Background and Objectives Thalassemia is the most prevalent hereditary anemia worldwide and in our country. Lifelong blood transfusion is currently the treatment protocol of this disease, but it has some side effects; one of them is alloimmunization. Alloantibodies produce antigens against RBCs which patients do not have. The antigens are transmitted via chronic blood transfusion gradually. One of these RBC antigens is antigen I, and the antibody against it is IgM class and cold antibody. Cold agglutinins are capable of agglutinating RBCs in low temperature and may result in hemolytic anemia which is known as cold agglutinin disease (CDA). In Iran, data regarding incidence of CAD are not available, but incidence of CAD in USA is approximately 1:300000. Case An 18-year-old man with b-thalassemia major referred to the hematology lab for periodic checking of blood smear. He had clumps of RBC and RBC indices were abnormal. In repeated examination, he was known to be asymptomatic for purpose of cold agglutinins.  Conclusions We do not have any records or information about cold agglutinins in thalassemic patients in Iran. This case was detected only through periodic laboratory tests without any special complications on the part of the patient.