Intercomparação e calibração de medidores de atividade utilizados em serviços de medicina nuclear

OBJETIVO: O objetivo deste trabalho foi estabelecer um padrão de trabalho para intercomparação e calibração de medidores de atividade (calibradores de dose) utilizados na maioria dos serviços de medicina nuclear, para determinação da atividade dos radionuclídeos administrados aos pacientes nos exames específicos ou nos procedimentos terapêuticos. MATERIAIS E MÉTODOS: Foi utilizado um calibrador de dose comercial, um conjunto de fontes radioativas padrões, além de seringas, frascos e ampolas contendo soluções de radionuclídeos utilizados em medicina nuclear. RESULTADOS: O calibrador de dose comercial foi calibrado com soluções de radionuclídeos utilizados em medicina nuclear. Os testes simples do instrumento, tais como o teste de linearidade e da variação da resposta com o volume da fonte a uma concentração de atividade constante, foram descritos e realizados. CONCLUSÃO: O instrumento estudado pode agora ser utilizado como sistema de referência para intercomparação e calibração de outros instrumentos medidores de atividade, como um método de controle da qualidade de calibradores de dose utilizados em serviços de medicina nuclear.

Proposta de método de inspeção de radioproteção aplicada em instalações de medicina nuclear

O objetivo principal deste trabalho é implantar um método de inspeção imparcial e eficiente, visando à utilização segura e correta das radiações ionizantes no campo da medicina nuclear. Este método de inspeção aqui proposto foi aplicado em 113 serviços de medicina nuclear do país, obedecendo a uma freqüência de análise bienal (1996, 1998, 2000 e 2002). Foram estabelecidos 82 itens de radioproteção com pesos para cada item, baseados nos fatores de risco, de acordo com as normas da Comissão Nacional de Energia Nuclear (CNEN) e as recomendações da Agência Internacional de Energia Atômica. Na análise do serviço, cada item de não conformidade com as normas da CNEN gerou uma irregularidade de radioproteção, com seu peso associado. O somatório dos pesos deu a cada serviço uma pontuação final que o classificou dentro de três faixas de valores que determinaram uma tomada de decisão por parte do órgão regulador e fiscalizador: funcionamento sem restrição - menor que 100 pontos; funcionamento com restrição - igual ou maior que 100 e menor que 300 pontos; funcionamento suspenso - igual ou maior que 300 pontos. Para o caso de irregularidades reincidentes, criou-se um fator multiplicativo para a pontuação. A reincidência em qualquer item de radioproteção (irregularidade) teve seu peso multiplicado por 2n, onde n era o número de vezes em que o item encontrava-se irregular. O estabelecimento prévio de itens de radioproteção, com atribuição de pesos para cada item, procurou minimizar os valores subjetivos e pessoais presentes no julgamento e na avaliação técnica das instituições inspecionadas.

Characterization of the HIV protease inhibitor Nelfinavir cellular targets

The HIV protease inhibitors (HIV-PIs) are among the most potent antiviral drugs for the HIV infection. Treatment of patients with HIV-PIs has been linked with development of side effects including dyslipidemia, lipodystrophy syndrome and cardiovascular complications. Moreover, these drugs have shown anti-tumoral activity in non-infected patients but little is known about the involved molecular mechanism for these off-target effects. Here we propose that the HIV-PI Nelfinavir could block a cellular protease thus causing the observed phenotypes. We firstly focus our attention on a cellular protein, DDI2, showing sequence and structural conservation with the HIV protease. We applied cellular and in vitro approaches to produce a correctly folded recombinant protein in order to investigate the presence of a proteolytic activity. Despite the fact that we could identify two techniques that can be applied to produce a folded recombinant DDI2, no proteolytic activity has been identified in the present study. However, we could observe that decreasing the DDI2 levels recapitulated some phenotype observed in presence of HIV-PIs, including the phosphorylation of the protein translation regulators eIF2a and eEF2. As an alternative approach to identify cellular targets for HIV-PIs, we applied a proteomic screening called Slice-SILAC. We focused our attention on the defective maturation of Lamin A, a member of the nuclear lamina, induced by the block of the cellular protease Zmpste24. We demonstrated that Nelfinavir induced accumulation of Prelamin A and nuclear shape defects and in addition caused presence of cytosolic DNA, probably due to TREX1 downregulation. We showed that these phenotypes correlated with activation of the AIM2 inflammasome and IL-lß release. These findings suggest that DDI2 and Zmpste24 are direct or indirect cellular targets for the HIV-PIs and indicate a possible role for these proteins in promoting off-target effects and anti¬tumoral activity observed in HIV-PI treated patients. -- Les inhibiteurs de la protéase du VIH (IP-VIH) sont parmi les médicaments antiviraux les plus efficaces pour l'infection par le VIH. Le traitement des patients avec les IP-VIH cause des effets secondaires comprenant la dyslipidémie, le syndrome de lipodystrophie et les complications cardio-vasculaires. De plus, ces médicaments ont montré une activité anti-tumorale chez les patients non infectés, toutefois le mécanisme moléculaire impliqué dans ces effets hors-cible reste inconnu. Nous proposons que l'IP-VIH Nelfinavir puisse bloquer une protéase cellulaire provoquant les phénotypes observés. De ce fait, nous avons concentré notre attention sur une protéine cellulaire, DDI2, qui possède une séquence et une structure proche de celle de la protéase du VIH. Nous avons appliqué des approches cellulaire et in vitro pour produire une protéine recombinante correctement repliée afin d'étudier son activité protéolytique. Malgré le fait que nous avons pu identifier deux techniques qui peuvent être appliquées pour produire une protéine DDI2 recombinante correctement repliée, aucune activité protéolytique n'a été identifiée dans la présente étude. De plus, nous avons pu observer que la réduction de DDI2 récapitule les phénotypes observé avec le IP-VIH, y compris les phosphorylations de eIF2a et eEF2, impliquées dans la régulation de la traduction protéique. Une approche alternative, appelée Slice-SILAC, a été utilisée afin d'identifier de nouvelles cibles cellulaires du Nelfinavir. Nous avons concentré notre attention sur la maturation défectueuse de la Lamine A, un membre de la lamine nucléaire, induite par l'inhibition de la protéase cellulaire Zmpste24. Nous avons démontré que le Nelfinavir induit une accumulation de Prélamine A déformant la membrane nucléaire et la présence d'ADN cytosolique, probablement en raison de la régulation négative de TREX1. Nous avons montré que ces phénotypes causent l'activation de l'inflammasome AIM2 et la sécrétion d'IL-lß. Ces résultats suggèrent que DDI2 et Zmpste24 sont des cibles cellulaires pour les IP-VIH et indiquent un possible rôle pour ces protéines dans l'apparition d'effets secondaires ainsi que dans l'activité anti-tumorale observée chez les patients traités avec les IP-VIH.

Neutron skin of 208Pb, nuclear symmetry energy, and the parity radius experiment

A precise determination of the neutron skin thickness of a heavy nucleus sets a basic constraint on the nuclear symmetry energy (the neutron skin thickness is the difference of the neutron and proton rms radii of the nucleus). The parity radius experiment (PREX) may achieve it by electroweak parity-violating electron scattering (PVES) on 208Pb. We investigate PVES in nuclear mean field approach to allow the accurate extraction of the neutron skin thickness of 208Pb from the parity-violating asymmetry probed in the experiment. We demonstrate a high linear correlation between the parity-violating asymmetry and the neutron skin thickness in successful mean field forces as the best means to constrain the neutron skin of 208Pb from PREX, without assumptions on the neutron density shape. Continuation of the experiment with higher precision in the parity-violating asymmetry is motivated since the present method can support it to constrain the density slope of the nuclear symmetry energy to new accuracy.

Proteasome inhibition reduces proliferation, collagen expression, and inflammatory cytokine production in nasal mucosa and polyp fibroblasts

Proteasome inhibitors, used in cancer treatment for their proapoptotic effects, have anti-inflammatory and antifibrotic effects on animal models of various inflammatory and fibrotic diseases. Their effects in cells from patients affected by either inflammatory or fibrotic diseases have been poorly investigated. Nasal polyposis is a chronic inflammatory disease of the sinus mucosa characterized by tissue inflammation and remodeling. We tested the hypothesis that proteasome inhibition of nasal polyp fibroblasts might reduce their proliferation and inflammatory and fibrotic response. Accordingly, we investigated the effect of the proteasome inhibitor Z-Leu-Leu-Leu-B(OH)2 (MG262) on cell viability and proliferation and on the production of collagen and inflammatory cytokines in nasal polyp and nasal mucosa fibroblasts obtained from surgery specimens. MG262 reduced the viability of nasal mucosa and polyp fibroblasts concentration- and time-dependently, with marked effects after 48 h of treatment. The proteasome inhibitor bortezomib provoked a similar effect. MG262-induced cell death involved loss of mitochondrial membrane potential, caspase-3 and poly(ADP-ribose) polymerase activation, induction of c-Jun phosphorylation, and mitogen-activated protein kinase phosphatase-1 expression. Low concentrations of MG262 provoked growth arrest, inhibited DNA replication and retinoblastoma phosphorylation, and increased expression of the cell cycle inhibitors p21 and p27. MG262 concentration-dependently inhibited basal and transforming growth factor-β-induced collagen mRNA expression and interleukin (IL)-1β-induced production of IL-6, IL-8, monocyte chemoattractant protein-1, regulated on activation normal T cell expressed and secreted, and granulocyte/macrophage colony-stimulating factor in both fibroblast types. MG262 inhibited IL-1β/tumor necrosis factor-α-induced activation of nuclear factor-κB. We conclude that noncytotoxic treatment with MG262 reduces the proliferative, fibrotic, and inflammatory response of nasal fibroblasts, whereas high MG262 concentrations induce apoptosis.

Proteasome inhibition reduces proliferation, collagen expression, and inflammatory cytokine production in nasal mucosa and polyp fibroblasts

Proteasome inhibitors, used in cancer treatment for their proapoptotic effects, have anti-inflammatory and antifibrotic effects on animal models of various inflammatory and fibrotic diseases. Their effects in cells from patients affected by either inflammatory or fibrotic diseases have been poorly investigated. Nasal polyposis is a chronic inflammatory disease of the sinus mucosa characterized by tissue inflammation and remodeling. We tested the hypothesis that proteasome inhibition of nasal polyp fibroblasts might reduce their proliferation and inflammatory and fibrotic response. Accordingly, we investigated the effect of the proteasome inhibitor Z-Leu-Leu-Leu-B(OH)2 (MG262) on cell viability and proliferation and on the production of collagen and inflammatory cytokines in nasal polyp and nasal mucosa fibroblasts obtained from surgery specimens. MG262 reduced the viability of nasal mucosa and polyp fibroblasts concentration- and time-dependently, with marked effects after 48 h of treatment. The proteasome inhibitor bortezomib provoked a similar effect. MG262-induced cell death involved loss of mitochondrial membrane potential, caspase-3 and poly(ADP-ribose) polymerase activation, induction of c-Jun phosphorylation, and mitogen-activated protein kinase phosphatase-1 expression. Low concentrations of MG262 provoked growth arrest, inhibited DNA replication and retinoblastoma phosphorylation, and increased expression of the cell cycle inhibitors p21 and p27. MG262 concentration-dependently inhibited basal and transforming growth factor-β-induced collagen mRNA expression and interleukin (IL)-1β-induced production of IL-6, IL-8, monocyte chemoattractant protein-1, regulated on activation normal T cell expressed and secreted, and granulocyte/macrophage colony-stimulating factor in both fibroblast types. MG262 inhibited IL-1β/tumor necrosis factor-α-induced activation of nuclear factor-κB. We conclude that noncytotoxic treatment with MG262 reduces the proliferative, fibrotic, and inflammatory response of nasal fibroblasts, whereas high MG262 concentrations induce apoptosis.

N-acetylcysteine treatment ameliorates the skeletal phenotype of a mouse model of diastrophic dysplasia.

Diastrophic dysplasia (DTD) is a recessive chondrodysplasia caused by mutations in SLC26A2, a cell membrane sulfate-chloride antiporter. Sulfate uptake impairment results in low cytosolic sulfate, leading to cartilage proteoglycan (PG) undersulfation. In this work, we used the dtd mouse model to study the role of N-acetyl-l-cysteine (NAC), a well-known drug with antioxidant properties, as an intracellular sulfate source for macromolecular sulfation. Because of the important pre-natal phase of skeletal development and growth, we administered 30 g/l NAC in the drinking water to pregnant mice to explore a possible transplacental effect on the fetuses. When cartilage PG sulfation was evaluated by high-performance liquid chromatography disaccharide analysis in dtd newborn mice, a marked increase in PG sulfation was observed in newborns from NAC-treated pregnancies when compared with the placebo group. Morphometric studies of the femur, tibia and ilium after skeletal staining with alcian blue and alizarin red indicated a partial rescue of abnormal bone morphology in dtd newborns from treated females, compared with pups from untreated females. The beneficial effect of increased macromolecular sulfation was confirmed by chondrocyte proliferation studies in cryosections of the tibial epiphysis by proliferating cell nuclear antigen immunohistochemistry: the percentage of proliferating cells, significantly reduced in the placebo group, reached normal values in dtd newborns from NAC-treated females. In conclusion, NAC is a useful source of sulfate for macromolecular sulfation in vivo when extracellular sulfate supply is reduced, confirming the potential of therapeutic approaches with thiol compounds to improve skeletal deformity and short stature in human DTD and related disorders.

Nuclear Factor I-C acts as a regulator of hepatocyte proliferation at the onset of liver regeneration.

BACKGROUND & AIMS: Knockout studies of the murine Nuclear Factor I-C (NFI-C) transcription factor revealed abnormal skin wound healing and growth of its appendages, suggesting a role in controlling cell proliferation in adult regenerative processes. Liver regeneration following partial hepatectomy (PH) is a well-established regenerative model whereby changes elicited in hepatocytes lead to their rapid and phased proliferation. Although NFI-C is highly expressed in the liver, no hepatic function was yet established for this transcription factor. This study aimed to determine whether NFI-C may play a role in hepatocyte proliferation and liver regeneration. METHODS: Liver regeneration and cell proliferation pathways following two-thirds PH were investigated in NFI-C knockout (ko) and wild-type (wt) mice. RESULTS: We show that the absence of NFI-C impaired hepatocyte proliferation because of plasminogen activator I (PAI-1) overexpression and the subsequent suppression of urokinase plasminogen activator (uPA) activity and hepatocyte growth factor (HGF) signalling, a potent hepatocyte mitogen. This indicated that NFI-C first acts to promote hepatocyte proliferation at the onset of liver regeneration in wt mice. The subsequent transient down regulation of NFI-C, as can be explained by a self-regulatory feedback loop with transforming growth factor beta 1 (TGF-ß1), may limit the number of hepatocytes entering the first wave of cell division and/or prevent late initiations of mitosis. CONCLUSION: NFI-C acts as a regulator of the phased hepatocyte proliferation during liver regeneration.

Análise dosimétrica de acompanhantes de pacientes de medicina nuclear internados em quarto terapêutico

OBJETIVO: Este trabalho avaliou doses recebidas por acompanhantes que compartilharam o mesmo quarto terapêutico com pacientes tratados com câncer de tireóide ingerindo iodo-131 (131I) e comparou os resultados obtidos aos limites determinados pelas normas brasileiras de radioproteção. MATERIAIS E MÉTODOS: Avaliaram-se seis grupos de pessoas. Cada grupo foi formado por um acompanhante e um paciente, que compartilharam o mesmo quarto terapêutico. Realizaram-se, também, 23 outros experimentos, sendo que nestes um simulador foi usado em substituição à segunda pessoa no quarto terapêutico. As atividades de 131I administradas aos pacientes foram de 3.700 MBq ou 5.550 MBq. Dosímetros termoluminescentes na forma de pó, fluoreto de lítio dopado com magnésio, titânio e sódio foram usados para a avaliação das doses. RESULTADOS: Os resultados mostraram que uma pessoa que compartilha o mesmo quarto terapêutico, durante dois dias, com um paciente tratado com 3.700 MBq ou 5.550 MBq de 131I, seguindo as orientações de radioproteção fornecidas pela equipe médica, é exposta a uma dose média de (0,51 ± 0,02) mSv, para um nível de confiança de 99%. CONCLUSÃO: De acordo com as normas brasileiras, do ponto de vista da radioproteção, não há impedimento acompanhar um paciente de medicina nuclear durante esse tratamento.

Levantamento do controle de qualidade de calibradores de dose de radiofármacos em serviços de medicina nuclear na cidade de São Paulo

OBJETIVO: Realizar levantamento sobre quais testes de controle de qualidade são realizados nos calibradores de dose dos serviços de medicina nuclear da cidade de São Paulo, SP. Estudar a exatidão das medições de atividade de sete calibradores de dose no Laboratório de Calibração de Instrumentos do Instituto de Pesquisas Energéticas e Nucleares, usando fontes de radionuclídeos importantes clinicamente. MATERIAIS E MÉTODOS: O levantamento sobre o controle de qualidade foi realizado a partir de questionários enviados aos serviços. Foram utilizados, no estudo de exatidão dos sete calibradores de dose, fontes de Ga-67, Tc-99m e Tl-201 e um instrumento padrão secundário. RESULTADOS: Os resultados do levantamento sobre os testes de controle de qualidade mostraram algumas impropriedades, por exemplo, a falta da realização diária do teste de reprodutibilidade por todos os serviços. Os resultados do teste de exatidão para os sete calibradores de dose estudados mostraram-se dentro do limite de aceitação da norma nacional (±10%). CONCLUSÃO: A situação com relação ao controle de qualidade de calibradores de dose é insatisfatória, de acordo com o pequeno número de serviços que participaram do levantamento. O estudo da exatidão em sete calibradores de dose não indicou falhas de desempenho e estabeleceu uma calibração desses instrumentos para as fontes utilizadas.

Taxation of nuclear waste in Spain

Law 15/2012 established in Spain four new environmental taxes and extended the scope objective excise duties on mineral oils to tax the use of natural gas and coal as sources of electricity. One of the newly created taxes falls on all electric power producers, and has as tax base the turnover. The second one tax hydropower production, and the other two fall on the nuclear industry. So, there are two new taxes in Spain on the production of electricity from nuclear sources. The first one is a tax on nuclear waste production; the second one is a tax on the storage of nuclear waste. However, these are not the only levies in the Spanish tax system affecting nuclear waste. At the State level there are already several charges on nuclear waste. At the regional level, on the other hand, two Autonomous Communities were taxing nuclear waste. The creation of these new State taxes will finish with the regional taxes, but the State will be oblige to compensate these regions for losing revenues. The purpose of this work is to carry out a critical analysis of the Spanish system of taxation on nuclear waste.