Portuguese Institute of Blood and Transplantation

Lisbon, Portugal

Portuguese Institute of Blood and Transplantation

Lisbon, Portugal
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Lourenco J.,University of Aveiro | Pereira R.,University of Aveiro | Pereira R.,University of Porto | Caetano T.,University of Aveiro | And 6 more authors.
Toxicology | Year: 2013

Environmental exposure to uranium and its daughter radionuclides, has been linked to several negative effects such as those related with important physiological processes, like hematopoiesis, and may also be associated with genotoxicity effects. Herein, genotoxic effects, immunotoxicity, trace elements and C reactive protein (CRP) analyses, were performed in peripheral blood samples collected from individuals of a population living near a deactivated uranium mine. C reactive protein analysis was performed to exclude candidates with active inflammatory processes from further evaluations. DNA damage and immunotoxicity (immunophenotyping and immune cell counts) were evaluated by comet assay and flow cytometry, respectively. Significant DNA damage was observed in the peripheral blood samples from volunteers living in the Cunha Baixa village. A significant decrease of NK and T lymphocytes counts were observed in the individuals from the Cunha Baixa village, when compared with individuals from the reference site. Uranium and manganese levels were significantly higher in the Cunha Baixa village inhabitants. On the other hand, zinc levels were significantly lower in those individuals when compared with the volunteers from the control village. Results suggest that inhabitants from Cunha Baixa have a higher risk of suffering from serious diseases such as cancer, since high DNA damages were observed in peripheral blood leukocytes and also decreased levels of NK and T cells, which play an essential role in the defense against tumor growth. © 2013 Elsevier Ireland Ltd.


Pereira P.,Portuguese Institute of Blood and Transplantation | Westgard J.O.,University of Wisconsin - Medical School | Encarnacao P.,Catholic University of Portugal | Seghatchian J.,International Consultancy in Blood Components Quality Safety Improvement
Transfusion and Apheresis Science | Year: 2015

The European Union regulation for blood establishments does not require the evaluation of measurement uncertainty in virology screening tests, which is required by ISO 15189 guideline following GUM principles. GUM modular approaches have been discussed by medical laboratory researchers but no consensus has been achieved regarding practical application. Meanwhile, the application of empirical approaches fulfilling GUM principles has gained support. Blood establishments' screening tests accredited by ISO 15189 need to select an appropriate model even GUM models are intended uniquely for quantitative examination procedures. Alternative (to GUM) models focused on probability have been proposed in medical laboratories' diagnostic tests. This article reviews, discusses and proposes models for diagnostic accuracy in blood establishments' screening tests. The output of these models is an alternative to VIM's measurement uncertainty concept. Example applications are provided for an anti-HCV test where calculations were performed using a commercial spreadsheet. The results show that these models satisfy ISO 15189 principles and that the estimation of clinical sensitivity, clinical specificity, binary results agreement and area under the ROC curve are alternatives to the measurement uncertainty concept. © 2014.


Pereira P.,Portuguese Institute of Blood and Transplantation | Westgard J.O.,University of Wisconsin - Medical School | Encarnacao P.,Catholic University of Portugal | Seghatchian J.,International Consultancy in Blood Components Quality Safety Improvement | de Sousa G.,Portuguese Institute of Blood and Transplantation
Transfusion and Apheresis Science | Year: 2015

The risk of uncertain results in infectious agents' tests is recognized in blood establishments, being particularly evident during the blood donor selection. The current risk-based approaches require risk assessment and "risk-based thinking". Accordingly, the blood establishment should consider the effect of uncertainty in all the technical decisions taken in a screening laboratory. Since the post-transfusion safety is one of the blood establishments' goals, the risk of post-transfusion infection should be evaluated and actions taken to decrease the chance of blood donations validation use false negative results. This article reviews and discusses the sources of uncertainty of infectious agents' reported results in blood establishments. It describes a set of sources of uncertainty that should be considered in screening immunoassay's decisions. The infectious agents' uncertainty concern is critical for reporting reliable results. © 2015 Elsevier Ltd.


Pereira P.,Portuguese Institute of Blood and Transplantation | Westgard J.O.,University of Wisconsin - Medical School | Encarnacao P.,Catholic University of Portugal | Seghatchian J.,International Consultancy in Blood Components Quality Safety Improvement | de Sousa G.,Portuguese Institute of Blood and Transplantation
Transfusion and Apheresis Science | Year: 2015

The screening laboratory has a critical role in the post-transfusion safety. The success of its targets and efficiency depends on the management system used. Even though the European Union directive 2002/98/EC requires a quality management system in blood establishments, its requirements for screening laboratories are generic. Complementary approaches are needed to implement a quality management system focused on screening laboratories.This article briefly discusses the current good manufacturing practices and good laboratory practices, as well as the trends in quality management system standards.ISO 9001 is widely accepted in some European Union blood establishments as the quality management standard, however this is not synonymous of its successful application. The ISO "risk-based thinking" is interrelated with the quality risk-management process of the EuBIS "Standards and criteria for the inspection of blood establishments". ISO 15189 should be the next step on the quality assurance of a screening laboratory, since it is focused on medical laboratory.To standardize the quality management systems in blood establishments' screening laboratories, new national and European claims focused on technical requirements following ISO 15189 is needed. © 2015 Elsevier Ltd.


PubMed | Catholic University of Portugal, International Consultancy in Blood Components Quality Safety Improvement, University of Wisconsin - Medical School and Portuguese Institute of Blood and Transplantation
Type: Journal Article | Journal: Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis | Year: 2015

The risk of uncertain results in infectious agents tests is recognized in blood establishments, being particularly evident during the blood donor selection. The current risk-based approaches require risk assessment and risk-based thinking. Accordingly, the blood establishment should consider the effect of uncertainty in all the technical decisions taken in a screening laboratory. Since the post-transfusion safety is one of the blood establishments goals, the risk of post-transfusion infection should be evaluated and actions taken to decrease the chance of blood donations validation use false negative results. This article reviews and discusses the sources of uncertainty of infectious agents reported results in blood establishments. It describes a set of sources of uncertainty that should be considered in screening immunoassays decisions. The infectious agents uncertainty concern is critical for reporting reliable results.


PubMed | Catholic University of Portugal, International Consultancy in Blood Components Quality Safety Improvement, University of Wisconsin - Medical School and Portuguese Institute of Blood and Transplantation
Type: Journal Article | Journal: Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis | Year: 2015

The screening laboratory has a critical role in the post-transfusion safety. The success of its targets and efficiency depends on the management system used. Even though the European Union directive 2002/98/EC requires a quality management system in blood establishments, its requirements for screening laboratories are generic. Complementary approaches are needed to implement a quality management system focused on screening laboratories. This article briefly discusses the current good manufacturing practices and good laboratory practices, as well as the trends in quality management system standards. ISO 9001 is widely accepted in some European Union blood establishments as the quality management standard, however this is not synonymous of its successful application. The ISO risk-based thinking is interrelated with the quality risk-management process of the EuBIS Standards and criteria for the inspection of blood establishments. ISO 15189 should be the next step on the quality assurance of a screening laboratory, since it is focused on medical laboratory. To standardize the quality management systems in blood establishments screening laboratories, new national and European claims focused on technical requirements following ISO 15189 is needed.


Henriques A.,University of Salamanca | Henriques A.,Portuguese Institute of Blood and Transplantation | Rodriguez-Caballero A.,University of Salamanca | Criado I.,University of Salamanca | And 8 more authors.
Haematologica | Year: 2014

Chronic antigen-stimulation has been recurrently involved in the earlier stages of monoclonal B-cell lymphocytosis, chronic lymphocytic leukemia and other B-cell chronic lymphoproliferative disorders. The expansion of two or more B-cell clones has frequently been reported in individuals with these conditions; potentially, such coexisting clones have a greater probability of interaction with common immunological determinants. Here, we analyzed the B-cell receptor repertoire and molecular profile, as well as the phenotypic, cytogenetic and hematologic features, of 228 chronic lymphocytic leukemia-like and non-chronic lymphocytic leukemia-like clones comparing multiclonal (n=85 clones from 41 cases) versus monoclonal (n=143 clones) monoclonal B-cell lymphocytosis, chronic lymphocytic leukemia and other B-cell chronic lymphoproliferative disorders. The B-cell receptor of B-cell clones from multiclonal cases showed a slightly higher degree of HCDR3 homology than B-cell clones from mono clonal cases, in association with unique hematologic (e.g. lower B-lymphocyte counts) and cytogenetic (e.g. lower frequency of cytogenetically altered clones) features usually related to earlier stages of the disease. Moreover, a subgroup of coexisting B-cell clones from individual multiclonal cases which were found to be phylogenetically related showed unique molecular and cytogenetic features: they more frequently shared IGHV3 gene usage, shorter HCDR3 sequences with a greater proportion of IGHV mutations and del(13q14.3), than other unrelated B-cell clones. These results would support the antigen-driven nature of such multiclonal B-cell expansions, with potential involvement of multiple antigens/epitopes. © Ferrata Storti Foundation.


PubMed | Sharif University of Technology, Iran Polymer And Petrochemical Institute, Biocant Biotechnology Innovation Center, Portuguese Institute of Blood and Transplantation and University of Coimbra
Type: Journal Article | Journal: ACS applied materials & interfaces | Year: 2016

Superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with antimicrobial agents are promising infection-targeted therapeutic platforms when coupled with external magnetic stimuli. These antimicrobial nanoparticles (NPs) may offer advantages in fighting intracellular pathogens as well as biomaterial-associated infections. This requires the development of NPs with high antimicrobial activity without interfering with the biology of mammalian cells. Here, we report the preparation of biocompatible antimicrobial SPION@gold core-shell NPs based on covalent immobilization of the antimicrobial peptide (AMP) cecropin melittin (CM) (the conjugate is named AMP-NP). The minimal inhibitory concentration (MIC) of the AMP-NP for Escherichia coli was 0.4 g/mL, 10-times lower than the MIC of soluble CM. The antimicrobial activity of CM depends on the length of the spacer between the CM and the NP. AMP-NPs are taken up by endothelial (between 60 and 170 pg of NPs per cell) and macrophage (between 18 and 36 pg of NPs per cell) cells and accumulate preferentially in endolysosomes. These NPs have no significant cytotoxic and pro-inflammatory activities for concentrations up to 200 g/mL (at least 100 times higher than the MIC of soluble CM). Our results in membrane models suggest that the selectivity of AMP-NPs for bacteria and not eukaryotic membranes is due to their membrane compositions. The AMP-NPs developed here open new opportunities for infection-site targeting.


PubMed | University of Coimbra and Portuguese Institute of Blood and Transplantation
Type: Journal Article | Journal: Clinical and experimental medicine | Year: 2016

In view of its heterogeneous presentation and unpredictable course, clinical management of systemic lupus erythematosus (SLE) is difficult. There is a need for biomarkers and diagnostic aids to monitor SLE disease activity and severity prior to, during and after treatment. We undertook this study to search for unique phenotypic patterns in each peripheral blood (PB) B cell subset, capable of distinguishing SLE patients with inactive disease versus SLE patients with active disease versus controls by using an automated population separator (APS) visualization strategy. PB was collected from 41 SLE patients and 28 age- and gender-matched controls. We analyzed the cell surface markers (in a tube CD20/CD27/CD19/CD45/CD38/CD81/BAFFR combination) expression on PB B cell subsets using principal component analysis, implemented in the APS software tool. Overall, our analysis indicates that active SLE can be distinguished from inactive SLE on the basis of a single tube analysis, focused on the decreased expression of CD38, CD81 and BAFFR in transitional B cells. The cluster analysis of immunophenotypic profiles of B cell subsets highlighted disease-specific abnormalities on transitional B cells that emerge as promising surrogate markers for disease activity. Further validation is needed with larger samples and prospective follow-up of patients.


PubMed | Portuguese Institute of Blood and Transplantation
Type: Journal Article | Journal: Inflammation research : official journal of the European Histamine Research Society ... [et al.] | Year: 2015

Here we evaluated whether allergic rhinitis to Dermatophagoides pteronyssinus induces alterations on circulating B cell subsets.Circulating B cell subsets and isotype expression on antigen-experienced B cells from allergic patients under conventional pharmacological treatment (NO-SIT, n = 15) and under subcutaneous immunotherapy (SCIT, n = 33), and non-allergic subjects (NC, n = 25) were analyzed by flow cytometry.In allergic patients, we found a significant decrease in IgM(+) and IgG(+) memory B cells and an increase in IgA(+) memory B cells. Additionally, the numbers of circulating IgA(+) plasmablasts in allergic patients were also increased, while those cells expressing IgM were reduced.Allergic patients have a disturbed B cell subsets distribution which seems to underlie rhinitis pathogenesis and remain unchanged after SCIT.

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