فصلنامه خون
پیاپی 6 (زمستان 1384)

Background and objectivesDiversity of heavy chain immunoglobulin (IgH) and T-cell receptor (TCR) molecules occures during B- and T-lymphocyte differentiation through the rearrangement of variable (V), diversity (D), junction (J) and constant (C) gene segments. Lymphoid leukemia cells are similar to normal precursors and have rearranged IgH, Igκ and TCR (cross-lineage rearrangement) genes which can be used as a marker of clonality and evaluation of minimal residual disease (MRD). The purpose of this study is to evaluate the pattern of TCR- δ/γ gene rearrangements using polymerase chain reaction (PCR) in B-precursor acute lymphoblastic leukemia (ALL) in Iranian children. Materials and Methods In our prospective study, bone marrow aspirates of 183 children with early diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 subjects with diagnosis of B-precursor ALLs were selected for study. Sixteen were excluded from our study due to various reasons including cellular degeneration. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, hyper-variable regions TCR- δ (Vδ2-Dδ3 and Dδ2-Dδ3) and TCR- γ(V γ; V γ I and V γ II) were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis (silver stain). The DNA sequences were compared and aligned to the homologous sequences of Gene Bank for confirmation. T-test, Mann whitney, Fisher exact test and Chi-square were used for data analysis. ResultsClonal rearrangement of TCR-γ (Vγ) and VγI/II were present in 79.3% and 64.9% of patients respectively and only 5% of cases showed biclonal pattern. The VγII rearrangement was the most common (46.8%) type in TCR-γ. 47 (45.2%) and 11 (16.6%) of patients had Vδ2- Dδ3 and Dδ2–Dδ3 partial gene rearrangements, respectively. Biclonal/oligoclonal patterns were present respectively in 27.7% and 4.3% of cases with Vδ2-Dδ3 rearrangement. Only one patient had biclonal Dδ2 Dδ3 rearrangement. ConclusionsClonal rearrangement of TCR-δ (Vδ2-Dδ3 and Dδ2-Dδ3) genes had similar pattern to other populations. Frequency of TCR- γ (V γ I and V γ II) rearrangements were slightly higher than previous reports. and in contrary to others except Brazilian report the V γ II rearrangement was the most common type. We found no significant correlation between presence of different types of rearrangements and quantitative variables and the only significat point was the reduction of Vδ2Dδ3 with increase in age. According to preliminary results, these clonal markers can be used in diagnosis and follow up of MRD.

Background and objectivesCancer patients due to bone marrow suppression, sepsis, and other relevant complications require appropriate blood components for transfusion. However, there are risks of transfusion reactions. These reactions are influenced by many factors varying across different geographic regions and medical centers. This study was performed to determine incidence of early transfusion reactions and their clinical symptoms and signs in cancer patients, and to calculate the correlation of these adverse reactions with some demographic data and some specifications of blood components. Materials and Methods In this descriptive study, 39 reactions from 4023 blood transfusion attempts were assessed. Patients were monitored for symptoms and changes in vital signs within 24 hours following transfusion. Data analysis was performed by descriptive and inferential statistics (X2 and ANOVA). ResultsThe majority of reactions belonged to platelets (56.43%) and packed cells (43.58%). The most common symptoms were shown to be rigors (2.72%), and fever (2.33%). The incidence rate was estimated to be 2.7% for FNHTR, and 2.1% for allergic reactions. Haptoglobin deficiency was found in 0.8% of FNHTR cases. A correlation was observed between sex and history of previous reactions (p=0.048, p=0.04, respectively) with blood transfusion reactions. ConclusionsThese findings indicated that incidence of blood transfusion reactions and clinical symptoms correlated with other studies. The incidence of reactions in women and individuals with previous history of repeated blood transfusions was shown to be greater. Transfusion of packed cell was associated with allergic reactions (p=0.04), and high platelet lifetime associated with pulmonary reactions (p=0.044).

Background and objectivesThe capsid or core Ag of Hepatitis C virus is a multifunctional protein which has the principal pathogenesis and diagnostic role in HCV related infections and most of these properties are attributed to the hydrophilic section (amino acids 2-122) of this protein. For different research and diagnostic applications, high amounts of this protein in pure and original form are required. So, the aim of this study was to clone the gene, optimize the expression condition, purify it in the original form, and immunologically characterize hydrophilic section of HCV Core Ag, expressed by T7-araBAD promoter system in E.coli. Materials and MethodsThe PCR amplified region corresponding to 2-122 section of this Ag from genotype Ib was cloned in pIVEX 2.3, a T7 promoter derived vector. The proper construct after digestional analysis and sequencing confirmations was transformed into BL21-AI E.coli, and protein expression under control of araBAD promoter by addition of 0.2% Arabinose was induced. ResultsAfter optimization of expression condition, purification of protein by NI-NTA agarose gel chromatography in native condition by immidazole yielded about 3.5mg/L of HCV core Ag. Immunological studies by western blotting through application of core specific mAbs and results of ELISA tests indicated that the protein is with desired immunological properties. ConclusionsAraBAD promoter can be perfectly utilized to produce the hydrophilic section of HCV core in high yields, and purification through NI-NTA in native condition may provide the antigen for different research and diagnostic applications.

Background and objectivesIn spite of major advances in the field of quality assurance in the process of collection, preparation and storage of platelets, bacterial infection following platelet transfusion remains a major problem in transfusion medicine. The present study was carried out in order to evaluate bacterial contamination of platelet concentrates collected at Tehran Regional Educational Blood Transfusion Center. Materials and MethodsBacterial growth of samples of platelet concentrates was studied in blood agar, EMB and thioglycollate broth after 48 hours at 37oC. The use of differentiation tests was made when any bacterial growth was observed. Simultaneously, the samples were also cultured in thioglycollate broth and studied for any turbidity or color change within 7 days. Any changes made the samples to be cultured in blood agar and EMB. Finally, the contamination rate and the ratio of contaminating bacteria were determined. Out of 7700 samples, three fourth (5775 samples) were taken from the cord and one fourth (1925) from both the bag and the cord. ResultsOut of 7700 samples of platelet concentrates studied, 14 (0.18%) were found positive for bacterial contamination. The contamination rate was estimated to be one in every 550 tested platelets (0.18%). Since in cases of blood bag contamination, the cord had been contaminated as well, there was then no difference on whether the sample was taken from the bag or cord. The bacteria identified were as follows: Staph. epidermidis (n=4), Staph. saprophyticus (n=2), Acinetobacter (n=5), Bacillus sp. (n=3). ConclusionsThe results show that screening platelet concentrates for bacterial contamination is necessary for blood transfusion centers and hospital blood banks.

Background and objectivesAppropriate diagnosis and proper treatment of the complications of head injury are the most common procedures encountered by neurosurgeons. Good management of the factors effective in the outcome of these cases, among which the derangements in the coagulation factors are one of the most common changes, can be one of the essential milestones in treatment of such victims. Materials and Methods The objectives of this study have been to find the derangements in the blood coagulating factors in the cases referred to the head trauma emergency care unit of Sina Hospital as a hospital-based study. We try to see if there is any correlation between these coagulopathies and the extracranial damages, brain damages in the CT-scans, and patients’ outcome. To our idea, it is for the first time that such a prospective study has been undertaken in Iran. ResultsAll the information are taken from the questionnaires addressed to 300 cases admitted to the emergency head trauma unit of Sina Hospital. Prothrombin time (PT), Thromboplastin time (PTT) and platelet counts, level of consciousness (GCS), associated extremities’ or visceral injuries, and findings in the X-ray films and CT scans, all have been included in this questionnaire. ConclusionsThe coagulopathies are much more prevalent in cases with severe head injury. The coagulopathies are also more prevalent in cases with remarkable brain damages in their imaging studies. They are much more prevalent in cases who are going to reach a bad outcome.

Background and objectivesBlood donors are deferred for numerous reasons. Some reasons for permanent deferral are intravenous drug use, male homosexual contact or certain positive test results. However, the majority of donor deferrals are short-term temporary deferrals (STTDs) that are resolved in a matter of days, weeks or months, thereafter donors would be considered again as eligible blood donors. The effect of STTDs on blood donor return rates and subsequent blood donations is studied. Materials and Methods The present study was historical cohort. Donors facing STTDs during the 15 Dec 1999 to 15 Mar 2000 were randomly computer-matched with non-deferred donors on the basis of donation date (case group: 804 donors; control group: 295 donors). Computer records were evaluated during the 3 years that followed (21 Mar 2000 to 21 Mar 2003) to determine donor return rates. Chi-square analysis was used for comparison. ResultsThe most common reasons for STTDs were elevated blood pressure, certain medication, bacterial infection, cold, and sore throat. Blood donor return rates were 32.4% (in the case of those deferred for cold /sore throat), 42% (those deferred for medication), 29.3% (those with hypertension), and 37.3% (for bacterial infection) over the 3 years that followed. Non deferred donors were a little more likely than donors with STTDs to return over these 3 years (36.6% vs. 34.8%; p=0.57), and non-deferred donors donated more whole blood units. ConclusionsDonors with STTDs have a negative impact on blood donor return rates. The blood center’s goal should be to keep donors in the donor pool. Measures to alleviate negative effects on donors with STTDs should be also taken.

Background and objectivesBeta-2 microglobulin (β 2MG) is the light chain of Histocompatibility–Class I human antigen and its normal range is <3mg/ml. β 2MG level in sera of hepatitis B patients increases. In Hepatitis infection the presentation of the viral antigen on the hepatocyte in the presence of Class I HLA antigen plays a major role in the elimination of the virus. Materials and Methods In this descriptive study, s β 2MG, HBsAg (by ELISA), and HBV DNA (by PCR) were evaluated in sera of 49 patients with hepatitis B and 35 subjects in control group. ResultsOur results showed HbsAg was positive in all patients. 29 of patients were HBV-DNA-PCR positive and 20 HBV-DNA-PCR negative. β 2MG in all subjects in control group was in normal range and in 34.7% of patients above normal limit. β 2MG in HBV-DNA-PCR positive patients was higher than HBV DNA PCR negative patients. Such differences were significant (p < 0.05). ConclusionsIt seems S β 2MG is a good marker for HBV replication and its absence may exclude HBV replication. The role of β 2MG in monitoring response to therapy needs to be further evaluated.

Background and objectivesTo prepare safe and adequate blood supply to meet patients’ needs and ensure a sufficient number of regular blood donors, knowledge about factors encouraging people to donate blood regularly is essential. Considering its importance, we aimed to survey positive and negative motivation for blood donation. Materials and Methods This survey was a cross-sectional research with a multiple questionnaire to study the positive and negative motivation toward blood donation in the first half of 1382 (according to Iranian calendar) in Shiraz Blood Transfusion Center. ResultsThis survey was conducted on subjects with positive motivation who referred to blood transfusion center at the mean age of 32.73 ± 9.6. 92.1% of the subjects were male, and 7.9% female, and 74.3% married. Positive motivation for blood donation included altruistic causes (65.3%), check-up purposes (12.9%), income-earning goals (8.9%), the positive impact of donation on health (8.9%), and curiosity about blood donation (4%). The mean age of the subjects with negative motivation was 25.22 ± 7.54. 22.7% were male, 77.3 female, and 67.7% single. The most important negative motivation was dizziness and faint (45.5%), and lack of time (4.5%). Negative motivation in women was statistically more significant than men (p<0.05). ConclusionsIn this study, the main positive motivation for blood donation came out of altruistic reasons, which are considered to be very positive. 12.9% of people just donated for check up that can endanger blood safety.The most important negative motivation was fear of dizziness and faint or disease as a result of donation. This can be removed through holding educational programs and providing sufficient care for donors.

Background and objectivesBombay phenotype donors do not have H Antigen on the surface of their red blood cells and have hemolytic anti-H in their sera; so, this group should receive autologous blood and donation of their blood as O blood group can lead to severe hemolysis in recipients. Therefore, it is important to detect them. Case A 46-year-old man living in Bushehr for many years has been recognized as O+ blood group. Preliminary diagnosis was made by the use of antibody screening agglutination test. In serologic studies, we detected high level of anti-H, anti-A, and anti-B. Direct coombs was also negative. Definite diagnosis was made with no red cell agglutination in the presence of anti-H. Complementary serologic studies showed that Kelly antigen was negative and Rh genotype was CD/cde. ConclusionsThis case increased the number of Bombay phenotypes in the world to 131 and in Iran to 11. It seems that this latter number is higher than official estimations of its prevalence (10-6). Therefore, the use of antibody screening test should be considered in screening blood donations.