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San Fedele Superiore, Italy

D'Alessandro A.,University of Tuscia | Grazzini G.,Italian National Blood Center | Giardina B.,University of Milan | Giardina B.,CNR Institute of Neuroscience | Zolla L.,University of Tuscia
Stem Cell Reviews and Reports

Umbilical-cord blood (UCB) has growingly become an accepted alternative source of hematopoietic stem cells for transplantation purposes. However, the low cell dose limit within a single unit is still an obstacle hindering the way of a broader diffusion. The real deal is the lack of knowledge about the molecular processes governing the events of expansion and differentiation of these cells. In order to fill this void, several studies were focused on the identification of the peculiar whole protein profile of UCB-derived hematopoietic stem cells. In this review article we provide a referenced list of overall proteins from UCB-derived hematopoietic stem and progenitor cells. This list has been elaborated for pathway and network analyses, along with GO term enrichment for biological and molecular functions, in order to individuate main classes of proteins governing functioning of these cells. From these analyses it seems to emerge a central role for heat shock proteins in immature hematopoietic stem cells. Their role might be relevant in protecting crucial transcription factors which drive proliferation and differentiation towards a specific lineage (e.g. erythroid, myeloid). Hereby we also stress the helpfulness of interactomics elaboration in providing a unified overview of independent proteomics data. It appears that maturation, other than representing a bottleneck to protein expression, could sculpt interaction maps via reducing complexity of immature interactomics profiles. © 2010 Springer Science+Business Media, LLC. Source

Liumbruno G.M.,Immunohematology and Trasfusion Medicine | Vaglio S.,Immunohematology and Trasfusion Medicine | Grazzini G.,Italian National Blood Center | Spahn D.R.,University of Zurich | Biancofiore G.,Liver Transplant Anesthesia and Critical Care
Minerva Anestesiologica

The overall use of allogeneic blood transfusions in clinical practice remains relatively high and still varies widely among centres and practitioners. Moreover, allogeneic blood transfusions have historically been linked with risks and complications: some of them (e.g. transfusion reactions and transmission of pathogens) have been largely mitigated through advancements in blood banking whereas some others (e.g. immunomodulation and transfusion-related acute lung injury) appear to have more subtle etiologies and are more difficult to tackle. Furthermore, blood transfusions are costly and the supply of blood is limited. Finally, evidence indicates that a great number of the critically ill patients who are being transfused today may not be having tangible benefits from the transfusion. Patient blood management is an evidence-based, multidisciplinary, multimodal, and patient-tailored approach aimed at reducing or eliminating the need for allogeneic transfusion by managing anaemia, perioperative blood conservation, surgical haemostasis, and blood as well as plasma-derivative drug use. From this point of view, the reduction of allogeneic blood usage is not an end in itself but a tool to achieve better patient clinical outcome. This article focuses on the three-pillar matrix of patient blood management where the understanding of basic physiology and pathophysiology is at the core of evidence-based approaches to optimizing erythropoiesis, minimising bleeding and tolerating anemia. Anesthesiologists and critical care physicians clearly have a key role in patient blood management programmes are and should incorporate its principles into clinical practice-based initiatives that improve patient safety and clinical outcomes. Copyright © 2015 Edizioni Minerva Medica. Source

D'Amici G.M.,University of Tuscia | Timperio A.M.,University of Tuscia | Gevi F.,University of Tuscia | Grazzini G.,Italian National Blood Center | Zolla L.,University of Tuscia

Factor VIII is an important glycoprotein involved in hemostasis. Insertion of expression vectors containing either the full-length cDNA sequence of human factor VIII (FLrFVIII) or B-domain deleted (BDDrFVIII) into mammalian cell lines results in the production of recombinant factor VIII (rFVIII) for therapeutic usage. Three commercially available rFVIII concentrates (Advate®, Helixate NexGen® and Refacto®), either FLrFVIII or BDDrFVIII, were investigated by 1- and 2-DE and MS. The objective of this study was to compare the heterogeneity and the high purity of both rFVIII preparations before and after thrombin digestion. In particular, the 2-D gel was optimized to better highlight the presence of contaminants and many unexpected proteins. Recombinant strategies consisting of insertion of expression vectors containing BDDrFVIII and FLrFVIII resulted in homogeneous and heterogeneous protein products, respectively, the latter consisting in a heterogeneous mixture of various B-domain-truncated forms of the molecule. Thrombin digestion of all the three rFVIII gave similar final products, plus one unexpected fragment of A2 domain missing 11 amino acids. Regarding the contaminants, Helixate NexGen® showed the presence of impurities, such as Hsp70 kDa, haptoglobin and proapolipoprotein; Refacto® showed glutathione S-transferase and β-lactamase, whereas Advate® apparently did not contain any contaminants. The proteomic approach will contribute to improving the quality assurance and manufacturing processes of rFVIII concentrates. In this view, the 2-DE is mandatory for revealing the presence of contaminants. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA. Source

Lombardini L.,Italian National Transplant Center | Bosi A.,University of Florence | Grosz S.,FH Campus Wien, University of Applied Sciences | Pamphilon D.,Jacie Executive Committee | And 6 more authors.
Vox Sanguinis

There have been great advances over the last decades in haematopoietic stem cell (HSC) transplantation, using either bone marrow, peripheral blood or cord blood-derived stem cells. The coming into force of the European legislation on tissues and cells and the consequent transposition of Directives into national laws have required the health authorities in the Member States (MS) and the scientific societies to review the transplantation activities to ensure the circulation of safe HSC products. Here, the regulatory inspection process performed by the Competent Authorities and the professional voluntary accreditation process of the Transplant Programmes active in Italy is compared. © 2013 International Society of Blood Transfusion. Source

Rebulla P.,Blood Transfusion Center | Pupella S.,National Institute of Health | Santodirocco M.,Puglia Cord Blood Bank | Greppi N.,Blood Transfusion Center | And 15 more authors.
Blood Transfusion

Background. In addition to a largely prevalent use for bleeding prophylaxis, platelet concentrates from adult blood have also been used for many years to prepare platelet gels for the repair of topical skin ulcers. Platelet gel can be obtained by activation of fresh, cryopreserved, autologous or allogeneic platelet concentrates with calcium gluconate, thrombin and/or batroxobin. The high content of tissue regenerative factors in cord blood platelets and the widespread availability of allogeneic cord blood units generously donated for haematopoietic transplant but unsuitable for this use solely because of low haematopoietic stem cell content prompted us to develop a national programme to standardise the production of allogeneic cryopreserved cord blood platelet concentrates (CBPC) suitable for later preparation of clinical-grade cord blood platelet gel. Materials and methods. Cord blood units collected at public banks with total nucleated cell counts <1.5×109, platelet count >150×109/L and volume >50 mL, underwent soft centrifugation within 48 hours of collection. Platelet-rich plasma was centrifuged at high speed to obtain a CBPC with target platelet concentration of 800-1,200×109/L, which was cryopreserved, without cryoprotectant, below -40 °C. Results. During 14 months, 13 banks produced 1,080 CBPC with mean (± standard deviation) volume of 11.4±4.4 mL and platelet concentration of 1,003±229×109/L. Total platelet count per CBPC was 11.3±4.9×109. Platelet recovery from cord blood was 47.7±17.8%. About one-third of cord blood units donated for haematopoietic transplant could meet the requirements for preparation of CBPC. The cost of preparation was € 160.92/CBPC. About 2 hours were needed for one technician to prepare four CBPC. Discussion. This study yielded valuable scientific and operational information regarding the development of clinical trials using allogeneic CBPC. Source

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