Dilemmas in considering β-thalassemia status in subjects with borderline HbA2 values: a pilot study in Eastern India

Interpreting hemoglobin disorders by high performance liquid chromatography can sometimes deceptive, especially with borderline HbA2 values. It is often problematic, especially in antenatal cases if the partner is a known thalassemia trait. We tested for underlying β-thalassemia mutations in 24 subjects with borderline HbA2 values (between 3.0%-4.0%). Amplification refractory mutation system-polymerase chain reaction was used to detect the five common Indian β-thalassemia mutations: [IVS-I-5 (G>C), Cod 15 (G-A), Cod 8/9 (+G), Fr. 41/42 (-TTCT) and Cod 26 (G-A)]. β-globin gene sequencing was performed if no mutation was detected. β-globin gene defect was not identified in any of the samples. There was no presence of any of the five common mutations in the small cohort. The average value of HbA2 in 24 normal samples was found to be 3.96. The average values for mean cell volume and mean cell hemoglobin (MCH) were found to be 82 and 28.8 pg respectively. Among these 24 normal samples, 13 had MCH below 27 pg and 11 had MCH above 27 pg. On the contrary, one thalassemic family was screened, in which the father of an HbE-β thalassemia patient was found to have HbA2 3.1, being a β-thalassemia carrier. Mutation analysis should be offered to all at-risk couples with borderline HbA2, especially those with values between 3.5% and 4.0% and microcytic hypochromic indices. As, cases with some specific mutational background or clinical condition shows abnormally low HbA2, so mutation screening should be performed in other partner if one partner found to be carrier or patient of thalassemia

Novel balanced chromosomal translocations in females with recurrent spontaneous abortions: Two case studies

Two couples with a history of recurrent pregnancy losses were referred to the Institute of Genetic Medicine and Genomic Science for cytogenetic evaluation. Chromosomal analysis of the phenotypically normal couples was done to investigate whether there are any new chromosomal abnormalities present in either of the couples caused recurrent pregnancy losses. Clinical and hormonal profile of the couples revealed normal parameters. The ultrasound scan of the females showed normal uterine and ovarian structures. Chromosomal analysis of the couples revealed normal 46, XY karyotypes in the both the male partners, and novel balanced reciprocal translocations 46, XX, t (5;8) (q35.3;q24.23) and 46, XX, t (4;13) (q12;q14) chromosomal constitutions in the female partners. Further, corroboration of the chromosome abnormalities was carried out by high resolution banding analysis. Unique and novel balanced reciprocal translocations were reported as an original investigation in two female partners from two different unrelated families both with the history of recurrent pregnancy losses

A FACS Based Case Study on Two HbE-β Thalassaemia Members of a Family, Having Similar Mutational Background

In this report we have tried to explain the reasons behind the difference in the pattern of transfusion requirement between two members of a family with similar β-globin mutation. The father and younger son both are HbE-β, but the father never had transfusion, whereas the younger son takes transfusion monthly. Mother and the elder son are HbEE without any history of transfusion. β-globin mutations of all family members were determined by ARMS-PCR. These were reconfirmed by direct sequencing of β-globin gene. Father and younger son were found to be Cod 26 (G-A)/IVS 1-5 (G-C), whereas mother and elder son were found to be Cod 26 (G-A)/Cod 26 (G-A). XmnI sequencing also revealed that all members of the family were CC. Then, flow cytometry study of red blood cells (RBCs) was performed to measure the oxidative stress of the RBCs. This study was also done on the light and dense fractions of the RBC population of the father and younger son. It was seen that the younger son suffers severe oxidative stress, which can be explained by his higher transfusion requirement. From our work, we have established the importance of taking oxidative stress of RBCs into consideration to explain the clinical manifestation and progression of haemoglobin related diseases like thalassaemia

Differential regulation of plasma proteins between members of a family with homozygous HbE and HbEβ-thalassemia

In this report we’ve compared the plasma protein profiles of 4 individuals in a family. Father and the younger son both are hemoglobin (Hb) Eβ-thalassemic {Cod 26 (G-A)/IVS 1- 5 (G-C)}, but the father never requires transfusion, whereas the younger son requires monthly blood transfusion. Mother and the elder son are HbEE {Cod 26 (G-A)/Cod 26 (GA)} without any history of transfusion. Proteomic study was done on the plasma fraction of the blood following ammonium sulphate precipitation. Proteins were separated by 2D-gel electrophoresis, expression of proteins compared by densitometry and proteins identified by tandem MALDI mass spectrometry. Proteins responsible in hemolysis, hypercoagulation and hemoglobin scavenging have shown differential regulation, establishing the relation between the differences in the levels of plasma proteins with the progression of the disease phenotype, manifested in the extent of transfusion dependence of the patient

Benzaldehyde-induced developmental genotoxicity triggers both neural and non-neuronal cells including the cells of immunity in Drosophila melanogaster

Although benzaldehyde, an aromatic aldehyde, has been declared safe for uses in food, conflicting reports exist regarding its genotoxic and cytotoxic potentials in organisms. Our present study is the first attempt to evaluate the effects of exposure of benzaldehyde on the entire course of development of a eukaryote model organism, Drosophila melanogaster. Total time required for the initial appearance of the third instar larvae, pupae and adults increased dose dependently with the increasing dietary concentration of benzaldehyde. Exposure of flies to each concentration of benzaldehyde caused dose-dependent and significant reductions in the population of pupae and young adults of the fly. Developmental inhibition was associated with dose dependent and significant structural aberrations of larval polytene chromosomes like ectopic pairing, inversion, fusion, etc., and deformities of hemocytes and neuroblasts and death of hemocytes. As much as 34% (SD ± 1.76)-52% (SD ± 1.7) and 18% (SD ± 2.5)-40% (SD ± 3.38) hemocytes and neuroblasts, respectively, underwent nuclear deformations in response to dietary exposures of flies to BA 100–1000 mg/l. Moreover, 16% (SD ± 0.52)-31% (SD ± 1.97) and 19% (SD ± 0.3)-33% (SD ± 1.78) hemocyte mortalities in response to BA 100–1000 mg/l were determined by two cell viability assays. Thus our study revealed that benzaldehyde was genotoxic to Drosophila melanogaster larvae that might be responsible for larval cell death and their subsequent developmental retardation. As this fly possesses substantial genetic homology with human, possibility of developmental inhibition of the later due to exposure of this chemical during pregnancy may not be ruled out