The Scientific Journal of Iranian Blood Transfusion
Year:2008 | Volume:4 | Issue:4|Papers Count:11

Background and Objectives: Globin chain synthesis and DNA analysis are among complementary tests for thalassemia diagnosis. Nowadays, DNA analysis is the only definitive method for diagnosis of suspected carriers. Despite the complexity of this heterogenic disease which is attributed to mutations in gene regulation sites or unknown mutations, globin chain synthesis has maintained its significant role in identifying different kinds of thalassemia. As a result, besides the routine application of this method , we decided to determine the ranges of a/b ratio values in different kinds of thalassemia.Materials and Methods: In this experimental study 214 cases were divided into the control (51 cases), minor b thalassemia (24 cases), mild a (a thal. 2) thalassemia (68 cases), severe a (a thal. 1) thalassemia (44 cases), Hemoglobin H disease (6 cases), silent G ( type II) thalassemia (14 cases), db thalassemia (5 cases), and adb thalassemia (2 cases) groups. CBC, hemoglobin electrophoresis using acetate cellulose paper in alkaline pH, hemoglobin A2 measurement by column chromatography, reticulocytes percentage, hemoglobin H, RBC morphology, and globin chain synthesis were performed on each group.Results: Significant differences were observed in mean values of RBC, hemoglobin, hematocrite, MCV, MCH, MCHC, a/b ratio in F and G thalassemia cases as compared with the control group. High prevalence of a thalassemia was observed among suspected individuals (55.2% of different kinds of F thalassemia vs. 9.8% of different kinds of atypic b thalassemia) as compared with atypic b thalassemia.Conclusions: The mean value of a/b ratio achieved in this study was similar to the others, but with a greater standard deviation. Because of this, there exists a wider range of a/b ratio. This width of range made overlaps in different and adjacent groups. Therefore, a/b ratio cannot be used by itself to firmly diagnose the type of thalassemia. As a result, for accurate diagnosis to be made, besides considering patient's ethnicity and clinical features, it is necessary to assess the results of CBC, hemoglobin electrophoresis pattern analysis, globin chain synthesis, familial tests, and DNA analysis.

Background and Objectives: Replacement of Val34Leu polymorphism in subunit A of coagulation factor XIII results in the replacement of Valine with Leucine in amino acid 34. As a result of this substitution, FXIII Val34Leu polymorphism acts as a factor for individual protection against thrombosis. For the first time in Iran, the prevalence of this polymorphism in patients with thrombotic events and in healthy individuals was determined and studied.Materials and Methods: The study was performed as a retrospective case-control one. 213 referral patients with thrombotic complications were admitted to IBTO Thrombosis and Hemostasis Laboratory. Their DNA was extracted using Qiagene kit. Using Polymerase Chain Reaction (PCR) and RFLP methods in the presence of restriction enzyme Cfo1, genotypes of FXIII Val34Leu polymorphism were identified. Statistical analysis was performed by SPSS software version 11.5 and confidence coefficient was 95%.Results: The prevalence of FXIII Val34Leu polymorphism in patients was 24.3% in patients and 37.4% in healthy individuals. The allele frequencies of leucine in cases and controls were 13.3% and 20.2% respectively. These results showed significant differences between the two groups.Conclusions: The present study demonstrates the association between FXIII Val34Leu polymorphism and protection against thrombotic disorders. The higher frequency of Leu allele and Val34Leu genotype in controls than in patients confirmed the results.

Background and Objectives: Beta major thalassemia is one of the most prevalent diseases in Iran where 20000 affected patients live; they should receive regular blood transfusion and desferal iron chelation to lead a normal quality of life. In this study, we evaluated transfusion and laboratory status of those patients referred to Mofid Children Hospital during the year of 2006-07.Materials and Methods: In this retrospective analytic-descriptive study, transfusion status of 121 patients referred to Mofid Children Hospital during 2006-07 was evaluated through their medical records. SPSS software and Mann-Whitney and Spearmann tests were used for data analysis.Results: 121 patients within the age range of 2-26 (average of 13±6.19) years, including 66 (54.5%) females and 55 males (45.5%) participated in the present study. Their pretransfusion hemoglobin level was in the range of 7.5-11.5 g/dl (average of 9.6±0.97 g/dl). In 82 patients (67.8%), hemoglobin level was above 9 g/dl. Serum ferritin level was 250-9500 ng/ml (average of 2133±1515 ng/ml). Serum folate was checked in 100 patients out of whom only 3% were below 3 ng/ml (normal reference range: 3-17.5 ng/ml). Positive HCV antibody and HCV RNA (PCR) was detected in 11 (9.1%) of patients who were between 16 to 26 years old, but only one (0.8%) had positive HBS antigen. HCV RNA (PCR) test was also positive in the patients. No patients were detected with HIV antibody. Serum ferritin level in hepatitis C patients fell within the range of 568-9500 ng/ml with the average of 3172±2772 ng/ml. Conclusions: It seems that beta major thalassemic patients in Mofid Children Hospital use appropriate blood transfusion, and iron chelation agents; they are also tested for transfusion transmitted infections. Precise control over blood transfusion and use of iron chelation agents, blood donor screening for transfusion transmitted infections especially hepatitis C, and vaccination against hepatitis B are important in improvement of the status of beta major thalassemics, but prevention is yet the most effective way to reduce the incidence disease and it's of complications.

Background and Objectives: Venous thrombosis is one of the important causes of morbidity and mortality all over the world. Elevated plasma homocysteine is known as a cause of vein morphology changes and endothelial dysfunction which lead to platelet activation, fibrinolysis inhibition and finally atherothrombosis. In this study, we evaluated the role of homocysteine in atherothrombosis as compared to the control group with no history of thrombosis.Materials and Methods: In this case control study‚ 100 patients with arterial thrombosis (54 men and 46 women) as the case group and 68 as control (40 men and 28 women) were involved. Blood samples were taken in the EDTA-located tube and transported to the laboratory for fasting plasma homocysteine to be measured by ELISA kits. Some data such as age, sex, thrombosis history, and familial thrombosis history were taken from the patients through a questionnaire. We measured fasting plasma homocysteine in both case and control groups by ELISA method. The statistical analysis was performed by SPSS statistical software using T-Test and Chi-square; odds ratio was also calculated.Results: The average rates of homocysteine in the case and control group were 23.85±18.4 and 11.48± 3.4 mmol/lit respectively showing statistical significance. The hyperhomocysteinemia frequency in the case group was 48%, whereas 17.6% in the control. A significant difference was also observed in the frequency of hyperhomocysteinemia between male (70.4%) and female (21.7%) in the case group. There was a moderate correlation between homocysteine level and age in the case group.Conclusions: According to the achieved odds ratio (2.72), hyperhomocysteinemia is an independent risk factor for thrombosis. It means that homocysteine measurement should be determined in thrombosis-affected or high risk patients. Dietary supplementation with low doses of folate and vitamin B12 should be considered in affected persons.

Background and Objectives: Bacterial contamination of blood products, especially platelets, may lead to bacterial sepsis or death and therefore is of concern. Many techniques have been explored to detect bacteria in blood products in order to prevent transfusion-related bacteria contamination and transmission. In the present study, four different methods were employed to detect 12 platelet units precontaminated with known bacteria.Materials and Methods: Ten units of platelet concentrates were inoculated at three levels (150, 15, and 1.5 CFU per ml) with Escherichia coli and Staphyococcus epidermidis. All of the platelet concentrates and two control units of platelet concentrates were stored at 20 to 24°C for five days. Every morning during storage, platelet concentrates were tested for platelet pH, plasma glucose, quantitative plate culture, and gram staining on platelet centrifuged smears.Results: Escherichia coli with 150 and 15 CFU per ml and staphylococcus epidermidis with 150 CFU per ml grew on culture medias after two days but staphylococcus epidermidis with 15 CFU per ml did after three days. The sensitivity rate of bacteria detection in platelet concentrates through gram staining was lower than quantitative culture. Despite lower plasma glucose level in platelet concentrates (as measured by hexokinase enzymatic method) inoculated with microbial staines, pH level in platelet concentrates (as measured by pH meter) contentiously increased during five days of storage.Conclusions: The sensitivity rate of bacterial detection in platelet concentrates through measuring extra cellular pH was estimated to be higher than that of plasma glucose, culture and gram staining methods.

Background and Objectives: Stem cells are defined with two main characteristics which are self renewal potential and pluripotency. It is of great importance to study expression of self renewal genes in different stem cells, because self renewal regulatory mechanisms may vary among different kinds of stem cell. Human umbilical cord stem cells have recently gained so much attention, because their use in cell therapy methods has several advantages. The main objective of the present study was to evaluate the expression of Oct-4, Sox-2, Nanog, Nucleostemin, Bmi-1 and Sox-9 self renewal genes in human umbilical cord vein mesenchymal stem cells.Materials and Methods: In this experimental study, umbilical cords were obtained in sterile condition and carried to the laboratory in HBSS buffer. Mesenchymal stem cells were harvested by collagenase treatment and cultured by means of their adhesiveness to plastic surfaces in DMEM medium supplemented with 15% FBS. To confirm the mesenchymal identity of these cells, expression of some surface markers including CD105, CD106, CD54, CD45, HLA-DR, CD166, and CD34 was analysed by means of flowcytometery. Finally, expression of self renewal genes was evaluated by RT-PCR and immunocytochemistry techniques.Results: Flowcytometry analysis of UBMCs revealed that the cells are positive for CD105 (98%) and negative for CD34 (0%), CD54 (1%), CD45 (0%), CD106 (0%), CD166 (0%), and HLA-DR (0%). Our results also revealed Oct-4, Nanog, Nucleostemin, Bmi-1 and Sox-9 expression in human umbilical cord vein mesenchymal stem cells, but not Sox-2 self renewal gene expression.Conclusions: The results showed that the mechanism which is known today for Oct-4, Nanog and Sox-2 function is not the only possible mechanism in which these three genes can play role to regulate self renewal potential of stem cells. Obtained results can offer that a combination of regulatory mechanisms which regulate self renewal of adult and embryonic stem cells, may be responsible to regulate self renewal of stem cells.

Background and Objectives: Malignant lymphoma may be very difficult to be diagnosed through routine histopathological methods because they may mimic reactive architecture or contain reactive infiltrates. Detection of the monoclonal lymphocyte population is one of the best methods of diagnosis since a monoclonal proliferation is strongly suggestive of neoplasia. By means of a PCR method it is possible to detect the immunoglobulin heavy chain (IgH) gene rearrangement and consequently the lymphocyte clonality.Materials and Methods: We evaluated formalin-fixed, paraffin-embedded biopsies, and bone marrow aspiration of 12 cases with non definite or suspicious histopathological diagnosis of non Hodgkin B cell lymphoma. After DNA extraction and quality control, semi-nested PCR was performed using consensus primers for amplification of CDR-3 region. PCR products were analyzed after heteroduplex analysis using polyacrylamide gel electrophoresis and silver staining.Results: 75% and 16.6% of patients showed clonal and polyclonal patterns respectively. One case showed weak monoclonal band in smear background and remained suspicious after duplicate PCR.Conclusions: Our findings are comparable with other international studies and support the concept that molecular techniques such as PCR provide a helpful approach in detection of monoclonal immunoglobulin rearrangements in malignant lymphoma. This is especially true for suspicious cases, but always in combination with clinical and histopathological information.

Background and Objectives: Beta thalassemia minor is a common form of microcytic anemia. Although it needs no treatment, prenatal screening of beta thalassemia followed by medical counseling for prevention of birth of children with beta thalassemia major is important before marriage. The specific criterion for beta thalassemia minor diagnosis is the elevated level of Hb-A2. Some discrimination indices helpful in detection of beta thalassemia minor were reported. The aim of this study was to define the sensitivity, specificity, and Youden's index for Kerman index I and II in screening of beta thalassemia minor.Materials and Methods: A total of 82 patients with microcytic anemia (MCV< 80 fl) including 42 with beta thalassemia minor and 40 with iron deficiency anemia were selected. Differential values of Kerman index I and II were calculated; then, the sensitivity, specificity and Youden's index were evaluated.Results: In screening of beta thalassemia trait, sensitivity, specificity, and Youden's index were observed to be 93%, 87%, and 80% for Kerman index I respectively; for Kerman index II these figures were calculated to be 76%, 90%, and 66% in order.Conclusions: Although none of the discrimination indices showed complete sensitivity and specificity, but Kerman index I with higher sensitivity and specificity came out to be more valid in screening of beta thalassemia minor.