Basilia Piraino

Oxidative damage and genotoxicity biomarkers in transfused and untransfused thalassemic subjects.

Chronic anemia and tissue hypoxia increase intestinal iron absorption and mitochondrial impairment in thalassemic patients. Regular blood transfusions improve hemoglobin levels but determine an iron overload that induces reactive oxygen species (ROS) overproduction. The aim of this study was to assess cellular oxidative damage by detection of ROS, lipid peroxidation, 8-oxo-dG, and mitochondrial transmembrane potential (Δψ(m)) in transfused and untransfused thalassemic patients. We have also evaluated genotoxicity by CBMN and comet assay. Our data show that ROS and lipid hydroperoxides are significantly higher in thalassemic patients than in controls, especially in untransfused thalassemia intermedia patients. Moreover, the latter have a significant decrease in Δψ(m) that highlights the energetic failure in hypoxic state and the ROS overproduction in the respiratory chain. 8-OHdG levels are higher in thalassemics than in controls, but do not differ significantly between the two patient groups. Both genotoxicity biomarkers highlight the mutagenic potential of hydroxyl radicals released by iron in the Fenton reaction. Values for percentage of DNA in the comet tail and micronuclei frequency, significantly higher in transfused patients, could also be due to active hepatitis C virus infection and to the many drug treatments. Our biomonitoring study confirms the oxidative damage in patients with thalassemia major and shows an unexpected cellular oxidative damage in untransfused thalassemic patients. In addition to iron overload, the results highlight the important role played by hypoxia-driven mitochondrial ROS overproduction in determining oxidative damage in β-thalassemias.

Cardiac complications and diabetes in thalassaemia major: a large historical multicentre study

The relationship between diabetes mellitus (DM) and cardiac complications has never been systematically studied in thalassaemia major (TM). We evaluated a large retrospective historical cohort of TM to determine whether DM is associated with a higher risk of heart complications. We compared 86 TM patients affected by DM with 709 TM patients without DM consecutively included in the Myocardial Iron Overload in Thalassaemia database where clinical/instrumental data are recorded from birth to the first cardiovascular magnetic resonance (CMR) exam. All of the cardiac events considered were developed after the DM diagnosis. In DM patients versus non‐DM patients we found a significantly higher frequency of cardiac complications (46·5% vs. 16·9%, P < 0·0001), heart failure (HF) (30·2% vs. 11·7%, P < 0·0001), hyperkinetic arrhythmias (18·6% vs. 5·5%, P < 0·0001) and myocardial fibrosis assessed by late gadolinium enhancement (29·9% vs. 18·4%, P = 0·008). TM patients with DM had a significantly higher risk of cardiac complications [odds ratio (OR) 2·84, P < 0·0001], HF (OR 2·32, P = 0·003), hyperkinetic arrhythmias (OR 2·21, P = 0·023) and myocardial fibrosis (OR 1·91, P = 0·021), also adjusting for the absence of myocardial iron overload assessed by T2* CMR and for the covariates (age and/or endocrine co‐morbidity). In conclusion, DM significantly increases the risk for cardiac complications, HF, hyperkinetic arrhythmias and myocardial fibrosis in TM patients.

Increased protein carbonyl groups in the serum of patients affected by thalassemia major

High oxidative stress status is known to be one of the most important factors determining cell injury in thalassemic patients and causing other serious medical complications, including a continuous proinflammatory status. The quantification of protein carbonyl groups in peripheral blood is widely used to measure the extent of oxidative modification. Thus, we measured serum concentrations of protein carbonyl groups in 30 patients affected by thalassemia major and in 15 healthy subjects. Strongly higher levels of protein carbonyl groups were measured in the blood from thalassemic patients than in that from healthy controls. Our findings evidence that thalassemic patients suffer from protein oxidative stress; the possibility of a role for carbonyl stress in the progression and severity of the disease needs further investigation.

Endocrinopathies, metabolic disorders, and iron overload in major and intermedia thalassemia: serum ferritin as diagnostic and predictive marker associated with liver and cardiac T2* MRI assessment.

Endocrinopathies and metabolic disorders‐characterized β thalassemic (βT) patients and the prevention and treatment of these comorbidities are important targets to be achieved. The aim of the study was to analyze the diagnostic and prognostic role of ferritin for endocrinopathies and metabolic disorders in βT patients. The ability of iron chelators to treat iron overload and to prevent or reverse metabolic disorders and endocrinopathies was also evaluated.

Hallerman-Streiff syndrome: patient with decreased GH and insulin-like growth factor-1.

We observed a 10-year-old boy with Hallerman– Streiff syndrome (HSS). The proband was the second child of healthy non-consanguineous parents. Family history was unremarkable. Pregnancy was uncomplicated and delivery at term with normal birth weight and length. There was mild psychomotor retardation. On physical examination, the child showed short stature (123 cm, 2.4 SD) with a distinctive triangular bird face. The head was brachycephalic with frontal bossing sparse curly and thin scalp hair. The eyebrows were hypoplastic. The palpebral fissures were short and downslanted with telecanthus and sparse eyelashes. The nasal bridge was narrow, the nose small with a rather beaked appearance and hypoplastic alae nasi. The philtrum was long and mouth small with thin lips. Palate was high arched. There was a distinct maxillary and mandibular hypoplasia. Malocclusion, prominent upper incisors, and dental crowding were quite remarkable. The ears were slightly low set with prominent ear lobes. Hands and feet were small with clinodactylous of 5th fingers. Distal interphalangeal joints of fingers and toes were enlarged. Laboratory tests, including extensive metabolic studies and karyotype yielded normal results. X-ray of left hand and wrist documented retarded bone age (8.5 years vs. 9.6 of chronologic age). Repeated insulin-like growth factor-1 (IGF-1) blood determinations disclosed constantly decreased levels in the range of 39–66.8 ng/ml (normal levels 128–458 ng/ml). The GH response to pharmacological stimuli yielded abnormal results with maximal levels of 5.8 ng/ ml after insulin stimulus and 8.5 ng/ml after clonidine stimulus (the maximal response must exceed 10 ng/ml). An ophthalmologic examination documented myopia and disclosed normal fundus oculi (Figs. 1–3). Short stature is a common feature in this syndrome, but to our knowledge, a deficiency of GH and IGF-1 has

Identification of α-thalassemia mutations in subjects from Eastern Sicily (Italy) with abnormal hematological indices and normal Hb A2

We analyzed the prevalence of α-thalassemia mutations in 298 subjects from Eastern Sicily (Italy) with reduced mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH), normal HbA2 and HbF, and normal serum iron. In 131 subjects (43.9%) we found six different genotypes of α-thalassemia: -α3.7/αα (36.6%), -α3.7/-α3.7 (27.5%), –MED/αα (10.0%), -α20.5/αα (9.1%), αHphIα/αα (8.4%), αHphIα/αHphIα (6.1%), and -α3.7/αHphIα (2.3%). Our data underline that in Eastern Sicily populations, the molecular screening of α-thalassemia mutations and/or deletions may be useful to better characterize the clinically asymptomatic subjects with a slightly reduced MCV and MCH and normal iron status.

HIF1α and Glut1 receptor in transfused and untransfused thalassemic patients

Keywords: hypoxia inducible factor 1; glucose transporter 1; hypoxia; transfusion; erythropoiesis

Thalassaemia major and infectious risk: High Mobility Group Box-1 represents a novel diagnostic and prognostic biomarker

High mobility group box ‐1 (HMGB1) represents a common causal agent for various types of diseases, including infective pathologies. This study aimed to investigate the role of HMGB1 in β‐thalassemia major (TM) by evaluating its diagnostic and prognostic role. Fifty‐one TM patients and 30 healthy subjects (HS) were enrolled. Receiver operating characteristics (ROC) analysis was employed to calculate the area under the curve (AUC) for HMGB1 to determine the best cut‐off values capable of identifying infectious episodes. Adjusted risk estimates for infective events were calculated using univariate followed by multivariate Cox proportional hazard regression analysis. Serum HMGB1 levels were higher in TM patients than in HS (14·6 ± 8·7 vs. 2·08 ± 0·9 ng/ml, P < 0·0001). Patients who underwent splenectomy were characterized by lower levels of HMGB1, when compared with patients with an intact spleen (10·2 ± 8 vs. 19·1 ± 7 ng/ml, P = 0·004). ROC analyses revealed an AUC for serum HMGB1 of 0·801, with a sensitivity and specificity of 92·3% and 68·2% to detect an infectious episode. Low HMGB1 levels predicted high risk of infective events (HR: 0·81; P = 0·006). HMGB1 represents a prognostic marker for TM patients and a predictive factor for infectious events.

Genotoxic effect of iron overload and disease complications in transfused β thalassaemic patients

In previously reported studies, we observed significantly high genotoxicity biomarkers in regularly transfused thalassaemic patients, thus, in this study, we better investigated the genotoxic effect of iron overload and of thalassaemia complications, including their drug treatments. The assessment was performed in 64 regularly transfused thalassaemic patients using cytokinesis-block micronucleus and comet assays. All patients were splenectomised and undergoing iron chelation therapy. To reduce hypoxia-induced oxidative damage, the patients with haemoglobin levels <9.5 g/dL were excluded. Serum concentrations of ferritin, iron, transferrin and the percentage of transferrin saturation, as well as cardiac and hepatic T2* magnetic resonance imaging, were considered to evaluate serum and organ siderosis.All genotoxic biomarkers significantly differed between patients and healthy subjects. Iron intake via blood transfusions was inversely related to percentage of DNA in tail. The disease complications affecting endpoints were active Hepatitis C virus infection, drug therapy for osteoporosis (i.e. bisphosphonates) and hormone replacement therapy for hypogonadism.The results, highlighting the combined effect of iron overload and, mainly, disease complications, including their respective pharmacological treatments, confirmed the increased cancer risk in thalassaemic patients.

Affective Control and Life Satisfaction in Thalassemics

Background. Thalassemia is a chronic disease that can lead to an impact on psychological functioning and social behavior of patients. However, still little is known about the specific psychological aspects of the disease, such as the degree of tension, life satisfaction and affective control, especially in adult patients.Aim. The purpose of this study is to investigate whether patients with thalassemia have specific psychological pattern relating to the dimensions of tension, satisfaction and quality of life, management of affection.Method. We evaluated 31 patients with thalassemia major and intermedia (19 women and 12 men) aged between 18 and 50 years (M = 34 + 16), belonging to the Complex Unit of Medical Genetics. For the evaluation were used the Profile of Mood States (POMS), the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Rorschach test.Results. The findings show an inverse relationship between the levels of self-reported tension and the affective control indicators at Rorschach. Life satisfaction, instead, seems to vary according to the severity of the disease - major vs. intermediate - and the type of therapy.Conclusions. An understanding of the psychological mechanisms involved in thalassemia, both self-reported and projective, can contribute to a wider patient take-over, by considering the subjective aspects related to the psychological and socio-emotional well-being, fundamental in the care compliance.