[Prevalence of common point mutations of alpha globin gene in Babol, Iran [2005-09]]

Alpha thalassemia is one of the most common hemoglobin disorders. Some combination of alpha globin gene mutations may cause HbH disease with severe anemia or intermediate thalassemia. genotype common deletions are routinely tested for suspicious alpha thalassemia couples but because of lack of information about the nature and frequency of point mutations and higher expenosor of sequencing, less attention was paid to them. This study was done to determine the prevalence of common point mutations of alpha globin gene in Babol, Iran. This descriptive study was carried out on DNA of 153 adult suspected to alpha-thalasemia with deleted alpha-golobolin gene referred to genetic laboratory in Babol, Iran during 2005-09. alpha 1 and alpha 2 genes were amplified by using specific biotinilated primers by PCR method. PCR products were assayed using 11 specific probs corresponding to common point mutations in alpha gene [C19, IVSI [-5nt], C59, Hb constant spring, Hb Icaria, Hb seal Rock, IVSI [148], C14, poly A [-2bp], poly A2, Poly A1] and fixed on byodine C membrabe. Hybridization between the probes and PCR products was visualized after a colorimetric reaction using of conjugated streptavidin peroxidase and TMB [tetra methyle Benzidine] and H[2]O[2]. The prevalence of point mutations in poly A2, 5nt, Hb constant spring and poly A1 were 28.75%, 14.38%, 7.84% and 2.61%, respectively. Point mutation in alpha globin genes was detected in%53.60 out of 153 adults suspected with alpha thalassemia without common deletion mutations

Effect of 8 weeks endurance training with two different durations on plasma HDL and ghrelin in male rats

Nesfatin-1, a novel anorexigenic protein derived from the Nucleobindin-2 [NUCB2] gene, is expressed in adipose tissue and is found in plasma. The purpose of this study was to examine the effect of an eight-week endurance training regimen on nesfatin gene expression and its concentration in the male rat liver. Eleven adult Wistar male rats were used. Animals were randomly divided into the training [TS, n=6] and control [CS, n=5] groups. Training groups were given exercise on a motor-driven treadmill [0% grade, 60 min, and 5 days/week for 8 weeks, 50-55%VO2max]. Samples of liver were excised and stored in liquid nitrogen to extract nesfatin-1 mRNA, and to determine its concentration and that of glycogen by RT-PCR, ELISA and colorimetric assay respectively. Although liver nesfatin mRNA expression and its concentration were increased, changes were not significant. Also liver glycogen concentration was significantly higher in trained rats compared to controls. The results of this research showed for the first time that nesfatin-1 is first expressed in the liver as a peripheral tissue and it then changes with endurance training. The insignificant variations of nesfatin-1 in the liver might be attributed to its role in energy balance. It seems that relative improvement in the liver's energy status is influenced by nesfatin gene expression, whereas as an indicator of source ATP, was lower in trained group compared to control group

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

[ frequency of SspI polymorphism in intron II of beta globin gene in Mazandaran province]

Due to the high annual birth rate of thalassemia major in our country, its prevention by prenatal diagnosis is of important priority. Gene mutation remains unknown in 10-20% of thalassemia trait people in Iran. In these cases, linkage analysis using polymorphic sites which are located near or within the gene is necessary to follow the mutant or the normal chromosome. SspI polymorphic site which is studied for the first time in Iran is located in the second intron of beta globin gene. The aim of this study was to determine the polymorphism frequency of this site in Mazandaran province. Peripheral blood of 211 thalassemia trait patients living in Mazandaran province was collected. After DNA extraction and amplification of the beta globin gene region containing the SspI polymorphic site, the effect of SspI restriction enzyme was evaluated on agarose gel. In 422 analyzed chromosomes, 20.6% were negative for SspI polymorphic site. Negative sites were almost equally associated with normal and mutant alleles [11.9% and 14.3% respectively]. SspI site analysis can be applied to follow the normal or mutant alleles of beta globin gene