HB Setif [A94(Gi)Asp+Tyr] in Malta

Since testing of newborn infants for abnormal hemoglobins (Hbs) was reinitiated in Malta in 1989, about 20,000 cord blood samples have been studied with an isoelectrofocusing (IEF) technique (1). Among these, 2% have an abnormal Hb, i.e. 1.8% have a y-globin variant, and 0.2% have an a-globin variant (2). P-Globin variants have been noted only sporadically. In a further survey for hemoglobinopathies in elderly members of the Maltese population, the presence of two a-globin variants that differed in amount and electrophoretic mobility, was noted (Fig. 1). One had a pl and peptide map consistent with that of Hb St. Luke’s [a95(G2)Pro+Arg] which occurs at levels of around 10% of total Hb in heterozygotes from Malta (3,4). The other had a slower electrophoretic mobility and was more abundant in the red cell lysate. It was present in the proband, a healthy 88-year-old male, and in his healthy 91 -year-old sister.peer-reviewe

Does quantitative heterogeneity of human fetal hemoglobin (Hb F) reveal friends or foes of KLF1 in globin gene switching?

The chemical heterogeneity of fetal hemoglobin (Hb F) due to variable ratios of the Gγ and Aγ globin subunits reflects genetic complexity because of common dimorphisms such as Hb F Sardegna (or Aγ75(E19) Ile>Thr; also known as AγT) in Caucasians, and common variants such the Gγ globin variant, Hb F Malta I (or Gγ117(G19) His>Arg) that is in tight linkage disequilibrium with the β globin variant Hb Valletta (or β87(F3) Thr>Pro) and is found in 1.8% of neonates from Malta.peer-reviewe

Georisks in the Mediterranean and their mitigation

An international scientific conference organised by the Seismic Monitoring and Research Unit, Department of Geoscience, Faculty of Science, Department of Civil and Structural Engineering and Department of Construction and Property Management, Faculty of the Built Environment, University of Malta.Part of the SIMIT project: Integrated civil protection system for the Italo-Maltese cross-border area. Italia-Malta Programme – Cohesion Policy 2007-2013This conference is one of the activities organised within the SIMIT strategic project (Integrated Cross-Border Italo-Maltese System of Civil Protection), Italia-Malta Operational Programme 2007 – 2013. SIMIT aims to establish a system of collaboration in Civil Protection procedures and data management between Sicilian and Maltese partners, so as to guarantee the safety and protection of the citizens and infrastructure of the cross-border area. It is led by the Department of Civil Protection of the Sicilian region, and has as other partners the Department of Civil Protection of Malta and the Universities of Palermo, Catania and Malta. SIMIT was launched in March 2013, and will come to a close in October 2015. Ever since the initial formulation of the project, it has been recognised that a state of national preparedness and correct strategies in the face of natural hazards cannot be truly effective without a sound scientific knowledge of the hazards and related risks. The University of Malta, together with colleagues from other Universities in the project, has been contributing mostly to the gathering and application of scientific knowledge, both in earthquake hazard as well as in building vulnerability. The issue of seismic hazard in the cross-border region has been identified as deserving foremost importance. South-East Sicily in particular has suffered on more than one occasion the effects of large devastating earthquakes. Malta, although fortunately more removed from the sources of such large earthquakes, has not been completely spared of their damaging effects. The drastic increase in the building density over recent decades has raised the level of awareness and concern of citizens and authorities about our vulnerability. These considerations have spurred scientists from the cross-border region to work together towards a deeper understanding of the underlying causes and nature of seismic and associated hazards, such as landslide and tsunami. The SIMIT project has provided us with the means of improving earthquake surveillance and analysis in the Sicily Channel and further afield in the Mediterranean, as well as with facilities to study the behaviour of our rocks and buildings during earthquake shaking. The role of the civil engineering community in this endeavour cannot be overstated, and this is reflected in the incorporation, from the beginning, of the civil engineering component in the SIMIT project. Constructing safer buildings is now accepted to be the major option towards human loss mitigation during strong earthquakes, and this project has provided us with a welcome opportunity for interaction between the two disciplines. Finally the role of the Civil Protection authorities must occupy a central position, as we recognize the importance of their prevention, coordination and intervention efforts, aided by the input of the scientific community. This conference brings together a diversity of geoscientists and engineers whose collaboration is the only way forward to tackling issues and strategies for risk mitigation. Moreover we welcome the contribution of participants from farther afield than the Central Mediterranean, so that their varied experience may enhance our efforts. We are proud to host the conference in the historic city of Valletta, in the heart of the Mediterranean, which also serves as a constant reminder of the responsibility of all regions to protect and conserve our collective heritage.peer-reviewe

Cancer-associated abdominal vein thrombosis

Cancer is associated with an increased risk of developing venous thromboembolism, due to its direct influence on the three pillars of Virchow’s triad (e.g., compression on the blood vessels by the tumour, blood vessels invasion, and cytokine release), together with the effect of exogenous factors (such as chemotherapy, radiotherapy, surgery). In cancer patients, the risk of thrombosis at unusual sites, such as splanchnic, ovarian and renal vein thrombosis, is also increased. Abdominal vein thromboses are frequently incidental findings on abdominal imaging performed as part of the diagnostic/staging workup or the follow-up care of malignancies. There is little evidence on the management of unusual site venous thromboembolism in cancer patients since there are only a few specific recommendations; thus, the management follows the general principles of the treatment of cancer-associated deep vein thrombosis and pulmonary embolism. This narrative review summarises the latest evidence on cancer-associated abdominal vein thrombosis, i.e., thrombosis of the splanchnic, ovarian and renal veins.peer-reviewe

Development of in vitro erythroblast cultures for transfusion purposes

Introduction: The demand for Red Blood Cell (RBC) transfusion is continuously increasing while the supply is not always sufficient. As there is no appropriate alternative to RBC transfusion, the in vitro manufacture of RBCs is a potential means to ensure an adequate and safe supply of blood products. Our previous studies developed an erythropoiesis model to culture human erythroblasts in vitro using media supplemented with Erythropoietin (Epo), Stem Cell Factor (SCF) and Dexamethasone (Dex) to allow survival, proliferation and self renewal capacity. Aim: To enhance erythroblast growth capacity in vitro by constitutive activation of SCF and Epo signaling and investigating the cooperative mechanisms with Dex resulting in induced self renewal potential. Methodology: Haematopoietic progenitors were isolated using Magnetic cell sorting. The CD34+ fraction was cultured in specific media and characterised by flow cytometry. The CD34- fraction was also kept in culture for erythroblast commitment and proliferation. The growth curves were calculated using the CASY cell counter and analyser. Cells were factor deprived for 4 hours and stimulated for 2 hours with Epo, SCF, Dex alone or selected combinations. Cells were harvested and the nuclear cell lysates isolated. The activation of the transcription factors NFkB, AP-1, CREB, NFAT and GR were investigated using Luminex technology. In addition, an electroporation protocol was optimised for expression studies. Currently Jak2 and cKit coding sequences are being amplified from cDNA samples. Results: The erythroblast culture originating from the CD34- fraction was improved by a co-culture of a stromal cell layer. This co-culture showed enhanced proliferation and retained a mean erythroblast diameter of 9.5µm. In the absence of the stromal layer an erythroblast diameter of 8.5µm was obtained, with continuous cell purification. The transcription factor multiplex profile of cultured erythroblasts showed a specific transcription activation upon the addition of Epo and Dex inducing CREB and NFAT activity. The CD34+ cell culture retained an immature cell morphology and the capacity to differentiate into erythroblasts upon culturing in a selective media (ESD). Conclusion: The identification of the co-culture system merits further investigation. Studying the mechanism of self renewal of erythroblasts and expression of Jak2 and cKit will assess the possibility of factor-independent growth of the blasts. The outcome could provide key insights into mass production of erythrocytes in culture which would eventually allow additional research for production of transfusion compatible erythrocyte units.peer-reviewe

Extracellular Hsp72 concentration relates to a minimum endogenous criteria during acute exercise-heat exposure

Extracellular heat-shock protein 72 (eHsp72) concentration increases during exercise-heat stress when conditions elicit physiological strain. Differences in severity of environmental and exercise stimuli have elicited varied response to stress. The present study aimed to quantify the extent of increased eHsp72 with increased exogenous heat stress, and determine related endogenous markers of strain in an exercise-heat model. Ten males cycled for 90 min at 50% O2peak in three conditions (TEMP, 20°C/63% RH; HOT, 30.2°C/51%RH; VHOT, 40.0°C/37%RH). Plasma was analysed for eHsp72 pre, immediately post and 24-h post each trial utilising a commercially available ELISA. Increased eHsp72 concentration was observed post VHOT trial (+172.4%) (P<0.05), but not TEMP (-1.9%) or HOT (+25.7%) conditions. eHsp72 returned to baseline values within 24hrs in all conditions. Changes were observed in rectal temperature (Trec), rate of Trec increase, area under the curve for Trec of 38.5°C and 39.0°C, duration Trec ≥ 38.5°C and ≥ 39.0°C, and change in muscle temperature, between VHOT, and TEMP and HOT, but not between TEMP and HOT. Each condition also elicited significantly increasing physiological strain, described by sweat rate, heart rate, physiological strain index, rating of perceived exertion and thermal sensation. Stepwise multiple regression reported rate of Trec increase and change in Trec to be predictors of increased eHsp72 concentration. Data suggests eHsp72 concentration increases once systemic temperature and sympathetic activity exceeds a minimum endogenous criteria elicited during VHOT conditions and is likely to be modulated by large, rapid changes in core temperature

Polycation-π Interactions Are a Driving Force for Molecular Recognition by an Intrinsically Disordered Oncoprotein Family

Molecular recognition by intrinsically disordered proteins (IDPs) commonly involves specific localized contacts and target-induced disorder to order transitions. However, some IDPs remain disordered in the bound state, a phenomenon coined "fuzziness", often characterized by IDP polyvalency, sequence-insensitivity and a dynamic ensemble of disordered bound-state conformations. Besides the above general features, specific biophysical models for fuzzy interactions are mostly lacking. The transcriptional activation domain of the Ewing's Sarcoma oncoprotein family (EAD) is an IDP that exhibits many features of fuzziness, with multiple EAD aromatic side chains driving molecular recognition. Considering the prevalent role of cation-π interactions at various protein-protein interfaces, we hypothesized that EAD-target binding involves polycation- π contacts between a disordered EAD and basic residues on the target. Herein we evaluated the polycation-π hypothesis via functional and theoretical interrogation of EAD variants. The experimental effects of a range of EAD sequence variations, including aromatic number, aromatic density and charge perturbations, all support the cation-π model. Moreover, the activity trends observed are well captured by a coarse-grained EAD chain model and a corresponding analytical model based on interaction between EAD aromatics and surface cations of a generic globular target. EAD-target binding, in the context of pathological Ewing's Sarcoma oncoproteins, is thus seen to be driven by a balance between EAD conformational entropy and favorable EAD-target cation-π contacts. Such a highly versatile mode of molecular recognition offers a general conceptual framework for promiscuous target recognition by polyvalent IDPs. © 2013 Song et al

Quantification of in-vivo : expression of the β-IVS-I-NT#6 thalassaemia mutation

A recently described β chain variant, Hb Valletta (or α2β287PRO; Felice .at aJ., BLOOD, 74, 7, suppl. 1, 141a, 1989) has been observed among 1.8% of the Maltese population among whom β+ & βo thal occur with a combined heterozygous incidence of 2.4 %. This provides an opportunity to quantify objectively, through the proportion of Hb A or of β(A) chains in double heterozygotes, the degree of functional deficit due to the β+ thal mutations which prevail in this area.peer-reviewe

Spectral counting assessment of protein dynamic range in cerebrospinal fluid following depletion with plasma-designed immunoaffinity columns

<p>Abstract</p> <p>Background</p> <p>In cerebrospinal fluid (CSF), which is a rich source of biomarkers for neurological diseases, identification of biomarkers requires methods that allow reproducible detection of low abundance proteins. It is therefore crucial to decrease dynamic range and improve assessment of protein abundance.</p> <p>Results</p> <p>We applied LC-MS/MS to compare the performance of two CSF enrichment techniques that immunodeplete either albumin alone (IgYHSA) or 14 high-abundance proteins (IgY14). In order to estimate dynamic range of proteins identified, we measured protein abundance with APEX spectral counting method.</p> <p>Both immunodepletion methods improved the number of low-abundance proteins detected (3-fold for IgYHSA, 4-fold for IgY14). The 10 most abundant proteins following immunodepletion accounted for 41% (IgY14) and 46% (IgYHSA) of CSF protein content, whereas they accounted for 64% in non-depleted samples, thus demonstrating significant enrichment of low-abundance proteins. Defined proteomics experiment metrics showed overall good reproducibility of the two immunodepletion methods and MS analysis. Moreover, offline peptide fractionation in IgYHSA sample allowed a 4-fold increase of proteins identified (520 vs. 131 without fractionation), without hindering reproducibility.</p> <p>Conclusions</p> <p>The novelty of this study was to show the advantages and drawbacks of these methods side-to-side. Taking into account the improved detection and potential loss of non-target proteins following extensive immunodepletion, it is concluded that both depletion methods combined with spectral counting may be of interest before further fractionation, when searching for CSF biomarkers. According to the reliable identification and quantitation obtained with APEX algorithm, it may be considered as a cheap and quick alternative to study sample proteomic content.</p

Evaluation of polygenic risk scores for breast and ovarian cancer risk prediction in BRCA1 and BRCA2 mutation carriers

Background: Genome-wide association studies (GWAS) have identified 94 common single-nucleotide polymorphisms (SNPs) associated with breast cancer (BC) risk and 18 associated with ovarian cancer (OC) risk. Several of these are also associated with risk of BC or OC for women who carry a pathogenic mutation in the high-risk BC and OC genes BRCA1 or BRCA2. The combined effects of these variants on BC or OC risk for BRCA1 and BRCA2 mutation carriers have not yet been assessed while their clinical management could benefit from improved personalized risk estimates. Methods: We constructed polygenic risk scores (PRS) using BC and OC susceptibility SNPs identified through population-based GWAS: for BC (overall, estrogen receptor [ER]-positive, and ER-negative) and for OC. Using data from 15 252 female BRCA1 and 8211 BRCA2 carriers, the association of each PRS with BC or OC risk was evaluated using a weighted cohort approach, with time to diagnosis as the outcome and estimation of the hazard ratios (HRs) per standard deviation increase in the PRS. Results: The PRS for ER-negative BC displayed the strongest association with BC risk in BRCA1 carriers (HR = 1.27, 95% confidence interval [CI] = 1.23 to 1.31, P = 8.2 x 10(53)). In BRCA2 carriers, the strongest association with BC risk was seen for the overall BC PRS (HR = 1.22, 95% CI = 1.17 to 1.28, P = 7.2 x 10(-20)). The OC PRS was strongly associated with OC risk for both BRCA1 and BRCA2 carriers. These translate to differences in absolute risks (more than 10% in each case) between the top and bottom deciles of the PRS distribution; for example, the OC risk was 6% by age 80 years for BRCA2 carriers at the 10th percentile of the OC PRS compared with 19% risk for those at the 90th percentile of PRS. Conclusions: BC and OC PRS are predictive of cancer risk in BRCA1 and BRCA2 carriers. Incorporation of the PRS into risk prediction models has promise to better inform decisions on cancer risk management