Β-thalassemia Major Patients Research Articles

Selective electrochemical sensing of hemoglobin from blood of β-thalassemia major patients by tellurium nanowires-graphene oxide modified electrode

Hemoglobin is an important part of the biological system that is involved in the transportation of oxygen and carbon dioxide. Abnormal level of hemoglobin is associated with different diseases such as β-thalassemia and anemia. An electrochemical sensor based on graphene oxide-tellurium nanowires (TeNWs/GO) is developed for the first time for electrochemical sensing of hemoglobin (Hb). Sensing parameters such as concentration, pH, scan rate, and effect of interferences are optimized. Linearity, detection, and quantification limits for TeNWs/GO-GCE are 0.996, 0.29 μM, and 0.967 μM respectively. Stability is measured by cyclic voltammetry (100 cycles) and chronoamperometry (15 h). TeNWs/GO-GCE quantitatively determines hemoglobin from blood samples of β-thalassemia major patients. To evaluate the relationship between RBC’s count and hemoglobin β-thalassemia major patients with the level of HbA, HbA2, and HbF statistical analysis is carried out. Shapiro-Wilk analysis shows a non-normal distribution and the results are calculated by Spearman correlation analysis. The results show that thalassemia patients have low levels of hemoglobin due to mutations in globin chains of hemoglobin.

Study of lipid profile in β thalassemia major pediatric patients with multiple blood transfusion and its correlation with serum ferritin level in tertiary care hospital in Kolkata

Background: β-Thalassemia major is associated with iron overload toxicity. This study was done to investigate lipid profile abnormality in β-Thalassemia major patients and to establish their correlation with serum ferritin. Another motto of this study was to signify deleterious effect of multiple blood transfusion and facilitate to understand how it will affect the lipid profile parameters changes so that we can also prevent early dyslipidemia induced complications like atherogenesis by early detection of lipid profile abnormality and also to establish derangement of lipid profile as an indirect evidence of iron overload.Methods: 102 nos. β-Thalassemia major paediatric patients with multiple blood transfusion (≥10) in age group of 6 to12 years admitted in pediatric ward of tertiary care hospital (NRSMCH) were enrolled. Serum ferritin level and lipid profile were investigated and their correlation were assessed.Results: Among 102 β-Thalassemia major pediatric (6 to 12 years) patients 55.88% were male and 66.67% were Hindu by religion and 69.61% children came from lower socio-economic class. The average value of number of previous blood transfusion was 42.79±7.510. The mean value of serum ferritin was 2263.53±833.904 ng/ml. The average value of serum total cholesterol, LDL and HDL were 153.84±5.428 mg/dl and 97.16±3.982 mg/dl and 29.45±4.445 mg/dl respectively and their correlation with serum ferritin were negative (r=-0.941 and r=-0.964 and r=-0.751 respectively). Average value of serum triglyceride and VLDL were 206.22±67.407 mg/dl and 36.16±3.385 mg/dl respectively and their correlation with serum ferritin were positive (r=+0.606 and r=+0.973 respectively).Conclusions: There was significant lipid profile derangement. As the serum ferritin level increases, serum total cholesterol, HDL, LDL decreases and serum triglyceride, VLDL level increases.

Novel retinal findings in β-thalassemia major: older age and higher ferritin level as the risk factors.

To investigate retinal changes in β-thalassemia major patients and identify their association with systemic risk factors. In this prospective study, 120 β-thalassemia major patients received complete ophthalmic examinations (best-corrected visual acuity, contrast sensitivity, color vision, and indirect ophthalmoscopy) and retinal imaging using color fundus photography and fundus autofluorescence imaging. Patients were grouped according to the presence of thalassemia-related retinal changes. The association between systemic risk factors (age, type and duration of iron chelator use, history of splenectomy, hemoglobin level, and ferritin level) and thalassemia-related retinal changes was investigated using logistic regression analysis. Thalassemia-related retinal changes were identified in 36.7% of patients. Several distinct retinal changes were observed, including retinal refractile bodies in 10% of patients and retinal hemorrhage in 5.8% of patients. Fundus autofluorescence imaging showed abnormal patterns in 36.3% of patients with thalassemia-related retinal changes and 18.4% of patients without thalassemia-related retinal changes. Age (odds ratio [OR] = 1.10, 95% confidence interval [CI] 1.03-1.18) and ferritin level (OR 1.16, 95% CI 1.01-1.33) were associated with thalassemia-related retinal changes. Novel retinal changes were observed in β-thalassemia major patients. This study identified older age and higher ferritin level as risk factors for thalassemia-related retinal changes.

A Pragmatic Scoring Tool to Predict Hydroxyurea Response Among β-Thalassemia Major Patients in Pakistan.

Despite high prevalence and incidence of β-thalassemia in Pakistan, there is very limited work on the use of hydroxyurea (HU) in thalassemia patients in the country. This is the first insight regarding genetic profiling of BCL11A and HU responses in Pakistani β-thalassemia. It correlates single-nucleotide polymorphisms on BCL11A (rs4671393, rs766432) and HBG2 (XmnI), age at first transfusion, and β-globin mutations with HU response in β-thalassemia major (BTM). Of 272 patients treated with HU, 98 were complete responders, 55 partial responders, and 119 nonresponders. Our analysis shows that HU response was significantly associated with patients having IVSI-1 or CD 30 mutation (P<0.001), age at first transfusion >1 year (P<0.001), and with the presence of XmnI polymorphism (P<0.001). The single-nucleotide polymorphisms of BCL11A were more prevalent among responders, but could not show significant association with HU response (P>0.05). Cumulative effect of all 5 predicting factors through simple binary scoring indicates that the likelihood of HU response increases with the number of primary and secondary genetic modifiers (P<0.001). Predictors scoring is a pragmatic tool to foresee HU response in patients with BTM. The authors recommend a score of ≥2 for starting HU therapy in Pakistani patients with BTM.

Evaluation of glycemic abnormalities in children and adolescents with \u03b2-thalassemia major

BackgroundThe quality of life of B-thalassemia major (β-TM) patients has improved with the use of frequent blood transfusions. However, this leads to chronic iron overload with its sequelae, as prediabetes and diabetes mellitus. This study aimed to assess insulin resistance and glucose abnormalities in a sample of B-thalassemia major patients in Benha, Egypt.ResultsThis case-control study included 40 B-thalassemia major patients on regular blood transfusion and iron chelation. Their ages ranged from 8 to 16 years, and 30 normal age and sex-matched controls. Thorough clinical examination was performed including weight (kg), height (m), body mass index (BMI) (kg/m2), and liver and spleen size. Laboratory investigations were done in the form of complete blood count, liver enzymes, serum ferritin, fasting plasma insulin, and fasting, and 2 h postprandial plasma glucose. Insulin resistance (IR) was calculated using the Homeostasis Model Assessment of insulin resistance (HOMA-IR) index. Insulin resistance was found in 27.5% of thalassemic patients; 18.2% of them had diabetes, 72.7% were prediabetics (with impaired fasting glycemia), and 9.1% had normal fasting and 2 h postprandial plasma glucose level. Insulin resistance increased significantly with increased blood transfusion duration, serum ferritin, liver enzymes, fasting plasma insulin, fasting plasma glucose, and 2 h postprandial plasma glucose (ROC). The curve analysis showed that the duration of blood transfusion, serum ferritin, fasting plasma insulin, fasting, and 2 h postprandial plasma glucose could significantly predict insulin resistance at a certain cut-off point.ConclusionOur data show that HOMA-IR can be used to detect insulin resistance in β-TM patients on long-term blood transfusions, especially patients with high serum ferritin and impaired liver enzymes.

Serum Zinc Level in β-Thalassemia Major: A Retrospective Study in Southwest Iran

Thalassemia, which is associated with compound complications, is the most common hereditary anemia in the world. The zinc level is different in patients with thalassemias. This study aimed to determine the zinc status and its relationship with demographic factors and chelation therapy in β-thalassemia major (β-TM) patients. This cross-sectional study was conducted on 102 β-thalassemia (β-thal), patients. Zinc level was evaluated in subjects using the atomic absorption method. The results were analyzed through descriptive statistics, χ 2 and Mann-Whitney U tests. Of 102 cases, 92 patients were eligible and evaluated. The mean age was 23 (minimum 11, maximum 43) years. Forty-six percent of cases were males and 54.0% were females. Of the 92 patients taking iron chelators, 29.3% used deferoxamine (DFO), 16.3% deferasirox (DFX), 20.6% DFO and deferiprone (DFP), and 33.8% DFO and DFP. All cases had zinc deficiency, 89.1% had zinc levels <40.0 mg/dL, and 10.9% with zinc levels >40.0 mg/dL. There was a significant relationship between gender and group with zinc levels greater than 40.0 mg/dL and those with less than 40.0 mg/dL. Zinc deficiency is highly prevalent among patients with thalassemia in the city of Yasuj, Iran. There was a significant relationship between zinc levels and gender although no significance was observed between zinc level and age, body mass index (BMI), ferritin, and chelation factors; it is recommended that these patients be periodically evaluated for zinc level. In case of a lack of laboratory evaluation, the use of prophylactic zinc supplementation should be considered for these patients.

Development of the Quality of Life Questionnaire (SF-36) for Patients with β-Thalassemia Major and β-Thalassemia Intermedia Based on Extended Rasch Analysis

The quality of life (QoL) questionnaire (SF-36) contains 36 questions in eight subscales. It requires much time to fill in by the respondent. The objective of this study was to use Rasch models to develop a questionnaire that brings the desired outcome of the QoL of people. Therefore, a new questionnaire was provided that is more motivating and time-saving for respondents than SF-36. The QoL of life assessment data of 325 β-thalassemia major (β-TM) and β-thalassemia intermedia (β-TI) patients in Kerman, Iran, was used as research data. In this study, the appropriate questions were classified in a limited dimensional framework using the exploratory factor analysis (EFA). The correctness of the factor structure was assessed by confirmatory factor analysis (CFA). The quality of the questions was evaluated by Rasch modeling [partial credit model (PCM)] and item analysis to ensure the validity and reliability of the questionnaire. Finally, the Pearson correlation coefficient was used to compare the new questionnaire with the previous one. This process resulted in the development of a new questionnaire with five subscales and 20 questions. The construct validity of the new questionnaire was good. The reliability index of the questionnaire was 0.75, and the Pearson correlation coefficient between the QoL scores gained from the previous and the new questionnaires was 0.93 that indicates the strength of the correlation. The use of Rasch analysis in this study resulted in the development of a new reliable and valid questionnaire.

A Novel Frameshift Mutation, Deletion of HBB:c.199_202delAAAG [Codon 66/67 (-AAAG)] in β-Thalassemia Major Patients from the Western Region of Uttar Pradesh, India.

PurposeBeta thalassemia is one of the most common inherited disorders in India with heterogenous clinical phenotypes from silent carrier to clinically severe ones. Our study aimed to characterize the mutation spectrum in thalassemia patients who are coming to the hospital for follow-up from the western region of Uttar Pradesh India.Patients and MethodsFor the study, a case series of the retrospective bi-centre study was conducted. The patients from two thalassemia centers in the major hospitals (LLRMC Meerut, and JNMC, Aligarh administered by the Ministry of Health and Family Welfare (MoHFW)) in the Western Uttar Pradesh, India were considered for the study. A total of 77 blood samples were obtained from individuals (both related and unrelated) diagnosed with β-thalassemia after their consent. After DNA extraction, HBB gene amplification, mutation-specific polymerase chain reaction and gene sequencing were carried out to analyze the mutations.ResultsIn this study, seven different types of mutations were reported for the first time in Western Uttar Pradesh, India. A novel frameshift mutation, deletion of 4 nucleotides Codon 66/67 (-AAAG) in exon 2 region, is reported for the first time. IVS 1–5 (G>C) and Codon 41/42 (-CTTT) are the most frequently reported mutations. The molecular spectrum for these two cases consists of 44 and 42 alleles out of 108 alleles, respectively.ConclusionA total of 108 β-thalassemia alleles were studied from 46 homozygous and 31 compound heterozygous patients. All the individuals were from 20 districts of the Western Uttar Pradesh, India.

Assessment of Quality of Life among Beta-Thalassemia Major Patients Attending the Hematology Outpatient Clinics at Cairo University Hospital

AIM: This paper aimed at assessing the quality of life (QoL) among beta (β)-thalassemia major patients using the short-form-36questionnaire (SF-36) and determining the factors associated with it.&#x0D; METHODS: A cross-sectional study was conducted among β-thalassemia major patients who were attending the hematology outpatient clinic at Cairo University Hospital using the consecutive sampling technique. Data were collected between October 2016 and March 2017. The QoL was assessed for patients aged ≥17 years. During the study period, a total number of 112 patients were included for participation.&#x0D; RESULTS: The mean age of the studied group was 18.32 ± 1.33 years. Most of the included patients (93.63%) had 1 monthly blood transfusion. The mean total score of SF-36 was 44.90 ± 7.54. Among the QoL domains of the studied patients, the “general health perception” domain was the most affected one with a mean score of (add the value of the score here), while the “vitality” domain was the least affected one. No statistically significant difference was reported between males and females regarding different QoL domains except for the “vitality” domain which mean score was significantly higher in males compared to females (p = 0.05). The age at onset of the disease and at first blood transfusion was the most documented factors to be positively correlated with the QoL among the studied patients.&#x0D; CONCLUSION: This study revealed that the QoL in thalassemia major patients is compromised. QoL assessment should be performed for all thalassemia patients to determine and implement the necessary interventions that focus on the affected domains.

Zinc Status in Beta-Thalassemia Major

BACKGROUND: Zinc is one of the most important minerals incorporated in the enzymes of the human body. Zinc may be deficient in patients with the β-thalassemia major with possible adverse outcomes.&#x0D; AIM: The purpose of this study was to assess the serum zinc status in β-thalassemia major patients in Duhok city.&#x0D; PATIENTS, MATERIAL, AND METHODS: In this case–control study, 70 children with β-thalassemia major (2–12 years) of both genders were enrolled and were matched with 70 apparently healthy children for age and sex. A venous blood sample was obtained from each child for the measurement of serum zinc and serum ferritin levels at Jin Center in Duhok City between January 1 and June 30, 2017.&#x0D; RESULTS: The mean serum zinc in the thalassemia patients (74.79 [±25.14] μg/dl) was significantly lower compared to the control group (93.61 [±15.12] μg/dl), (p = 0.0001). The serum zinc was not significantly different in thalassemia patients in terms of age, disease onset, gender, height, weight, body mass index, amount of blood transfusion, and type of chelation. There was a statistically significant correlation between serum zinc levels with a serum ferritin level of patients.&#x0D; CONCLUSION: The study showed that thalassemia patients have significantly lower serum levels of zinc with no relation to medical factors.

Molecular and genetic characterization of hepatitis B virus among multitransfused thalassaemia patients in Islamabad, Pakistan.

Background:Hepatitis B virus (HBV) is the aetiological agent of transfusion-transmitted hepatitis globally. Beta thalassaemia major individuals are at greater risk of contracting HBV infection due to multiple blood transfusions required for the medical management of these patients. Based on HBV genetic variability, it is divided into 10 genotypes. The determination of HBV genotypes has significant implications for clinical management and treatment regimens.Aim:This study was performed to assess the HBV epidemiology and circulating genotypes in multi-transfused β-thalassemia major patients with the aim to be considered while formulating the treatment pattern taking into account particular needs of thalassaemia patients.Materials and Methods:This study was performed from September 2018 to June 2019, at the Department of Pathology and Transfusion Medicine, Shaheed Zulfiqar Ali Bhutto (SZAB) Medical University, Islamabad. A total of 2,260 thalassaemia patients were enrolled in the study. The study was endorsed by the Ethics Committee of the SZAB Medical University, Islamabad. The samples were serologically screened for HBsAg on the LIAISON® XL Murex HBsAg Quant assay (DiaSorin S.p.A., Italy) a chemiluminescence based immunoassay (CLIA). HBV quantitative PCR kit was used to measure the HBV DNA in serum samples. The HBV genotypes were determined using universal primers targeting the P1 and S1 region amplification.Results:Of 2,260 thalassaemia patients, 64.6% were males while 35.4% were females. The HBsAg was identified in 98 individuals (4.33%). The PCR analysis was done for these 98 patients and in this cohort, genotype D was 59.18% (n = 58), genotype A was 21.42% (n = 21) while genotype C was 19.38% (n = 19).Conclusion:The determination of HBV genotypes in the multi-transfused patients is key to the effective management of chronic HBV patients as the severity and course of the disease is dependent on a specific type of genotypes. Quality assured screening of donated blood will prevent the incidence of HBV in thalassaemia patients.

Skewed intracellular cytokine production of iNKT cells toward Th2-related responses in alloimmunized thalassemia patients

IntroductionRed blood cell alloimmunization is a challenging issue in thalassemia patients. Several studies have investigated the role of different immune system compartment in alloimmunization, but the exact mechanism remains unclear. Considering the immunoregulatory function of iNKT cells and their subsets, in this study, we evaluated the possible role of these cells in alloimmunization status of thalassemia patients. Methods78 β-thalassemia major patients (41 alloimmunized and 37 non-alloimmunized) and 17 healthy controls were engaged in this study. Mononuclear cells were isolated from peripheral blood samples and stimulated for cytokine production. Samples were subjected to flow cytometry for enumeration of iNKT cells and characterized based on their cytokine production pattern. Finally, the results correlated with alloimmunization status, clinical and laboratory data. ResultsResults demonstrated that the number of iNKT, iNKT+IFN-ɤ+, and iNKT+IL-4+ cells in thalassemia group was significantly higher than healthy controls while no significant change was observed in the number of these cells between alloimmunized and non-alloimmunized thalassemia patients. Interestingly, the ratio of iNKT+IL-4+: iNKT+IFN-γ+ cells in alloimmunized thalassemia group represent a considerable increase in comparison to both non-alloimmunized thalassemia group and healthy controls. However, evaluating this value in non-alloimmunized group represents an approximately equal ratio of 0.94, which was almost similar to this ratio in the control group (0.99).Conclusion.Our results illustrated a noteworthy imbalance in the ratio of iNKT cell subsets in favour of IL-4 producing iNKT cells in alloimmunized thalassemia patients. Regarding the role of IL-4 in stimulating the Th2-related immune responses, this imbalance could consider as a possible mechanism in alloantibody responses of thalassemia patients.

Relation of Serum Ferritin Level with Serum Hepcidin and Fucose Levels in Children with β-Thalassemia Major

The aim of this study was to assess serum ferritin, hepcidin, L-fucose, and protein binding fucose levels in β-thalassemia major (β-TM) patients and to correlate the serum ferritin level with hepcidin and fucose levels. A total 70 (26 males and 44 females) children with β-TM, ages ranging from 5 to 16 years (mean age 8.3 ± 2.7 years) and 50 (25 males and 25 females) apparently healthy subjects with matching age and sex were included as a control group. An especially designed questionnaire was used to collect age, gender, body mass index (BMI), hemoglobin (Hb), serum ferritin, hepcidin-25 peptide, α-L-fucose, and protein binding fucose (PBF) levels. β-Thalassemia major patients had significantly (p < 0.05) higher serum ferritin, fucose and PBF levels, but the serum hepcidin level was significantly (p < 0.05) lower when compared to the controls, and their levels were affected by the gender of the β-TM patients, as it was significantly (p < 0.05) higher in female in comparison to male patients. There was no significant (p > 0.05) correlation between serum ferritin with hepcidin and fucose levels as a marker of iron overload in β-TM. The regulation of hepcidin, and L-fucose levels in patients with β-TM is more affected by erythropoeitic activity than by iron overload, as there was no significant correlation between serum ferritin with hepcidin and fucose levels as a marker of iron overload in β-TM.

Role of Complement Regulatory Proteins (CD55, CD59, and CD35) on Red Blood Cells of &amp;lt;i&amp;gt;β&amp;lt;/i&amp;gt;-Thalassaemia Patients

Background: β-thalassaemia (β-Thal) is an inherited chronic haemolytic anaemia resulting from absent or low level of synthesis of β-globin chains of haemoglobin A in erythropoietic cells. The complement system is an important part of innate immune response that may be implicated in red blood cell (RBC) lysis. Mammalian cells are provided with surface bound complement regulatory proteins (MCRPs) that regulate the activation of complement cascade, thus protecting them from uncontrolled complement-mediated lysis. Objective is to evaluate the role of complement regulatory proteins (CD55, CD59, and CD35) on red blood cells, and to explain the pathogenesis of anaemia in β-thalassemia major. Methods: This case-control study enrolled 74 β thalassemia major patients who were compared with 40 age and sex matched controls. We performed expression of CD55, CD59, and CD35 on RBCs using flow cytometry. Results: CD55 levels of β-thalassemia major patients (79.78% ± 18.54%) were significantly decreased compared to healthy controls (99.45% ± 0.59%) (P β-thalassemia major patients (97.76% ± 1.72%) were significantly lower than in the controls (99.75% ± 0.36%) (P β-thalassemia major patients (4.30% ± 4.66%) than in the control group (19.40% ± 10.90%) (P Conclusion: β-thalassemia major patients suffer from increased haemolysis and a consequent increase in their demand for blood transfusion. Complement-mediated haemolysis was shown in our study by decreased expression of CD55, CD59, and CD35 in β-thalassemia major patients. This allows complement deposition on RBCs and enhances or accelerates their lysis.

Characterization of Iron Complexes in the Red Blood Cells of β-thalassaemia patients using 57Fe Mössbauer Spectroscopy

Red Blood Cells (RBCs) samples from healthy volunteers, β-thalassaemia major and β-thalassaemia intermedia patients were studied by means of 57Fe Mössbauer Spectroscopy (MS). The MS spectra from the RBCs of the thalassaemia patients showed the existence of significant amounts of ferritin-like iron, and in particular in the 82% of the β-thalassaemia intermedia ones. Also, β-thalassaemia intermedia patients were found to have higher Liver Iron Concentration (LIC) when compared to the β-thalassaemia major patients, as assessed by MRI exams, even though they had on an average about half the serum ferritin levels.

Relation of hepcidin gene expression in blood mononuclear cells with iron overload severity among β-thalassemia major patients

Iron overload is the main cause of morbidity and mortality in β-thalassemia major patients, and cardiac iron overload is the most common reason for death in these transfusion-dependent patients. Hepcidin, a liver-derived peptide hormone, plays a key role in plasma iron levels regulation by controlling two main stages, digestive iron absorption in enterocytes, and iron recycling in macrophages. Although hepcidin is mainly secreted from hepatocytes in the liver, it is also synthesized from mononuclear cells consisting of monocytes and lymphocytes. Binding of this molecule to ferroportin, a specific cellular exporter of iron, leads to degradation of the ligand-receptor complex, which reduces the iron overload by lowering the amounts of iron released into the plasma. Likewise, the same mechanism has been proved to be true for lymphocyte-drived hepcidin.The expression levels of hepcidin mRNA were evaluated using quantitative real time PCR (qRT-PCR) in 50 β-thalassemia major patients, as well as 25 healthy volunteers as the group of control. There was a significantly positive correlation between the cardiac iron concentration, showed by higher T2 values, and hepcidin levels in the patients (p = 0.028; r = 0.311). However, hepcidin expression levels did not significantly correlate with ferritin and liver iron concentrations.Hepcidin can act as a beneficial marker to determine iron overload degrees, particularly in the heart, in β-thalassemia major patients and be used as a logical therapeutic agent for treatment of β-thalassemia disorders.

Genotyping of blood groups in alloimmunized patients with β-thalassemia major by T-ARMS-PCR and multiplex-aso-pcr

IntroductionBeta-thalassemia major is a severe hemolytic anemia requiring life-long blood transfusion. Planned random donor blood transfusion is associated with alloimmunization against incompatible antigens. Determination of the minor blood group systems phenotype or genotype, and administration of the compatible blood components can significantly reduce the rate of alloimmunization.The present study aimed to determine the prevalence of alloimmunization, and genotype/phenotype characteristics of the minor blood groups systems in patients with β-thalassemia major. Material and methodsThis study was conducted on 1147 β-thalassemia major patients. Initially, antibody screening and antibody identification were performed. Then, phenotyping and genotyping for the Rh, Kell, Kidd, and Duffy blood groups were done in alloimmunized patients using monoclonal antibodies and Multiplex-Allele Specific Oligonucleotide-Polymerase Chain Reaction (Multiplex-ASO-PCR) and Tetra-primer amplification refractory mutation system–PCR (T-ARMS-PCR), respectively. Any phenotype/genotype discrepancy was assessed by direct sequencing. ResultsNinety-seven (8.5 %) out of 1147 patients had alloantibodies against the minor blood group antigens (44 males, 45.4 %, and 53 female, 54.6 %). The most common alloantibodies were against the RH (n: 47, 48.5 %), and the Kell (n: 23, 23.7 %) blood groups systems. Twenty-three (2.1 %) genotype/phenotype discrepancies out of 1067 tests, including 9 in the Rh (9.3 %), 8 in Duffy (34.8 %), and 6 in Kidd (26.1 %) blood groups were detected. No discrepancy was found in the Kell blood group system. Direct sequencing revealed that the results of molecular methods were correct. ConclusionMultiplex-ASO-PCR and T-ARMS-PCR molecular methods are fast, reliable and cost-benefit molecular methods for the minor blood group genotyping in multi-transfused β-thalassemia major patients.

Detection of Parvovirus B19 in B-thalassemia Major Patients by Serological and Molecular Method

Introduction: Because to the tropism of human parvovirus B19 to erythroid progenitor cells, infectionin patients with an underlying hemolytic disorder such as Beta thalassemia major leads to suppression oferythropoiesis, referred to as reticulocytopenia,which could be life threatening, The aim of the study was todetermine the rate of occurrence of parvovirus (B19) in beta thalassemia major patient by using real timePCR, Study the correlation between B19 virus and patient’s descriptive data and the correlation betweenB19 viral load and IgG titer.Method: This case-control study was done to detect the presence of parvovirus B19 DNA in plasma samplesand anti-IgG ELISA of patients with beta thalassemia major. The population consisted of 75 patients withbeta-thalassemia major who attended the Aban Al-balady hospital in in the City of Baghdad and 75 healthypeople as a control.Results: The prevalence of parvovirus B19 in our study population was 28 (37.3%) were positive for B19DNA by quantitative polymerase chain reaction in which highly significant different P value (&lt; 0.0001),Results of enzyme linked immunosorbent assays showed that IgG antibodies was positive in 16 (21.3%)compared with control group which was 7 (9.3%). According to statistical analysis the difference was inborderline between case and control group in prevalence of B19 virus. There was significant (P= 0.039)association correlation between B19 DNA and anti-IgG positivity, but were significant in WBC (P&lt;0.0001).Conclusion: In study, B19 infections were discovered in patients with beta thalassemia major. Screening ofsuch risky groups will significantly reduce the incidence and prevalence of B19 infection.

Conformational changes of β-thalassemia major hemoglobin and oxidative status of plasma after in vitro exposure to extremely low-frequency electromagnetic fields: An artificial neural network analysis

ABSTRACT Electromagnetic fields (EMF) can generate reactive oxygen species and induce oxidative modifications. We investigated the effects of extremely low-frequency electromagnetic fields (ELF-EMF) on oxidative status of plasma and erythrocytes in β-thalassemia major patients and design artificial neural networks (ANN) for evaluating the oxyHb concentration. Blood samples were obtained from age and sex-matched healthy donors (n = 12) and major β-thalassemia patients (n = 12) and subjected to 0.5 and 1 mT and 50 Hz of EMF. Plasma oxidative status was estimated after 1 and 2 h exposure to ELE-EMF. Structural changes of plasma proteins were investigated by Native PAGE and SDS-PAGE. Moreover; multilayer perceptron (MLP) method was applied for designing a feed forward ANN model to predict the impact of these oxidative and antioxidative parameters on oxyHb concentration. Two hour exposure to ELF-EMF induced significant oxidative changes on major β-thalassemia samplesElectrophoretic profiles showed two high molecular weight (HMW) protein aggregates in plasma samples from healthy donors and major β-thalassemia patients. According to our ANN design, the main predictors of oxyHb concentration were optical density of Hb at 542, 340, 569, 630, 577, and 420 nm and metHb and hemichrome (HC) concentration. Accuracy of the proposed ANN model was shown by predicted by observed chart (y = 1.3 + 0.96x, R2 = 0.942), sum of squares errors (SSR), and relative errors (RE). Our results showed the detailed effects of ELF-EMF on Hb structure and oxidative balance of plasma in major β-thalassemia patients and significance of ANN analysis during normal and pathologic conditions.

Analysis of determinant factors of liver fibrosis progression in ex-thalassemic patients.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) potentially renders thalassemia patients disease-free with presumably cessation of associated complications. This study analyzes the liver fibrosis status and the determinants of its progression in ex-thalassemic patients. The liver fibrosis status of 108 pediatric transfusion-dependent β-thalassemia major patients was evaluated before and one year after allo-HSCT using transient elastography (TE). All patients achieved normal hematopoiesis. In univariate analyses, not in all, but in patients developing significant post-HSCT iron overload or hepatic graft-versus-host disease (GvHD), as well as recipients of bone marrow stem cells (BMSC), significant TE increment occurred. In multivariable analyses, through a model with large effect size (Adj.R2 = 26%, F(3,104) = 13.53, P < 0.001), post-HSCT serum ferritin and hepatic GvHD were ascertained as independent determinants of significant TE increase, and the effect of stem cell graft source approached the level of significance. Excluding the patients with intermediate/high Lucarelli risk classes, the TE increase was significantly greater only in BMSC recipients (P = 0.033). Although the risk impact of allograft source on liver fibrosis progression requires further evaluation; hepatic status of ex-thalassemic patients can be preserved after HSCT, if hepatic GvHD is controlled and adequate post-transplantation iron depletion is ensured.