[Molecular evaluation of hemoglobin D mutations in Mazandaran province, Iran]

Hemoglobinopathies are among the most prevalent genetic disorders worldwide, and occur as a result of mutations in the gene involved in synthesizing hemoglobin chains. By now more than 1000 defects in hemoglobin chains are discovered. Hemoglobin D [Hb D] is one of these disorders, identified by a single nucleotide mutation on codon 121 of beta globin chain. This study was carried out to evaluate Hb D mutations through molecular methods in Mazandaran province of Iran. This descriptive laboratory study was done on 70 patients with an electrophoresis band in hemoglobin-S zone in Mazandaran province of Iran during 2010-11. Capillary zone electrophoresis was done to find out Hb D in 51 patients. Subsequently, PCR-RFLP was performed to evaluate the samples at molecular level. Molecular investigation revealed all cases are carriers of hemoglobin D-Punjab. Two patients were shown to be homozygote carriers of the abnormal gene. This study showed all Hb D affected patients were carriers of Hb D Punjab

[Prevalence of hemoglobinopathies in premarriage individuals referred to Babolsar, Iran [2006-09]]

According to world health organization statistics, at least 5.2% of world population is carrier for a main hemoglobin disorder. Previous reports showed that more than 10% of people are carrier for beta-thalassemia Northern Iran. This study was done to determine the prevalance of hemoglobinopathies in premarriage individuals referred to Babolsar, Iran. This descriptive study was carried out on 8500 individuals [4200 women and 4300 men] whome were attended the thalassemia counseling program in Babolsar, North of Iran during 2006-09. After performing the CBC test, for those MCV and MCH were less than 80 and 27 respectively, Hemoglobin A2 was evaluated. Subjects whome were volunteers for more comprehensive tests, basic and acidic electrophoresis and genetic tests were applied, subsequently. 1200 [14.11%] subjects had low hematological indexes. 474 [5.57%] subjects had high HbA2 and were classified as beta-thalassemia carriers and 726 [8.54%] had normal HbA2 level and were classified as alpha-thalassemia carriers. 6 [1.2%] subjects were identified with HbF level more than 10 and were identified as carriers for beta-gene cluster deletion carrier. Also, 16 [3.2%] individuals had HbE, 16 [3.2%] had HbS, 4 had HbD and 4 had HbH [0.33% in 1200 and 0.047% in 8500 subjects]. Genetic study of 317 individuals for beta carriers and 145 subjects for alpha-carriers showed IVSII-1G>A [74.5%] in beta-globin and single gene deletion of 3.7 [47.5%] in alpha-globin genes were the most frequent mutations. This study showed that carriers for alpha-thalassemia [8.5%] are more frequent compared with beta-thalassemia [5.57%]. Also other hemoglobin variants included HbS, HbE, HbD or different beta-gene cluster deletions in the region are considerable and should be screened

study of relationships between beta-globin gene mutations in beta-thalassaemia patients, in response to hydroxyurea treatment

Beta-thalassaemia is the most frequent inherited disorder in the world, especially in Iran and Mazandaran Province. It is caused by mulation in beta-globin gene on chromosome 11 with more than 150 different mulations causing beta-thalassaemia, has been identified in the beta-globin gene to date. Hydroxyurea, is one of the drugs used in Thalassemia patient's treatment, however, it is not effective in all patients. The mechanisms of the hydroxyuea effect in not clear yet. This study compared different beta-globin gene mutations in beta-thalassaemia patients who were referred to the Thalassemia Research Center in Sari in two groups, good responder and non-responder, to the hydroxyurea. This was a case-control study, comparing two groups of 30 thalassaemic patients who received hydroxyurea. Two groups were included, 30 good responders to hydroxyurea treatment [control] and 30 who did not respond to the treatment [case]. First, DNA was extracted from peripheral blood. Then, two different methods for mutation detection were used. In the Thalassaemia Research Center in Sari, mutations in 60 patients were identified using ARMS-PCR. Also the results were confirmed in Genetic laboratory of Amirkola, using two mutation detection methods, reverse-dot blot hybridization and ARMS-PCR. In the group of good responder [control], the average patient's age were 28/1 +/- 7/78 years, and the average age at the onset of blood transfusion was reported to be 8/5 +/- 8/56 year. In this group, the mean comparison of the hemoglobin level and red blood size [MCV] prior and after drug consumption were statistically significant. In the group of non-responder [case], the mean age was 21.3 +/- 6.43, the mean age starting blood transfusions was 3.3 +/- 3.75, and the mean of drug consumption was 2.3 +/- 0.8 months. From the mutations identified, IVSII-1G>A was the most common type in both case and control group, while of 30 of control group, 22 individuals were homozygous, and 7 individuals were heterozygous for this mutation [frequency% 42.5]. For the 30 case patients, 11 individuals were homozygous, while 11 were heterozygous [frequency% 27.5]. Comparison between two groups, case and control group, were statistically significance [P < 0.008]. The correlations of IVSII-1G>A mutation in good responder patients to hydroxyurea as compared to the non responder group, is significant and similar to the previous findings

Phenotypic study and molecular analysis of von Willebrand type 1 patients

Von willebrand disease [VWD] is the most common bleeding disorder caused by von willbrand factor [VWF] deficiency and autosomal dominance inheritance pattern. It is divided into three types, one and three [quantitative] and type 2 [qualitative]. Type one is known with mild bleeding symptoms in comparison with type 3. The aim of this study was to determine the mutations responsible for the type one VWD. DNA was extracted from affected members of the family with type 1 VWD and the von willebrand gene was amplified using 63 different PCR, Then PCR fragments were analyzed using CSGE gel electrophoresis, and the fragments with the extra bands were analyzed using DNA sequencing. The average of von willebrand antigen and VWF activity were found 35.18 and 31.4 unit/deciliter in 23 patients from 6 families, respectively. Mutations were found in 4 families from 6. Argenine 1205 to histidine in 2 families and two new mutations including glysine 19 to arginine in exon 2 and a nucleotide change 2821 guanine to adenine [G>A] in intron 21 splice site in two other families. No mutation was found in two other families.This study showed that different kind of mutations causing von willebrand disease in different families