The Dually Acylated NH2-terminal Domain of Gi1α Is Sufficient to Target a Green Fluorescent Protein Reporter to Caveolin-enriched Plasma Membrane Domains: PALMITOYLATION OF CAVEOLIN-1 IS REQUIRED FOR THE RECOGNITION OF DUALLY ACYLATED G-PROTEIN α SUBUNITS IN VIVO

Here we investigate the molecular mechanisms that govern the targeting of G-protein α subunits to the plasma membrane. For this purpose, we used Gi1α as a model dually acylated G-protein. We fused full-length Gi1α or its extreme NH2-terminal domain (residues 1–32 or 1–122) to green fluorescent protein (GFP) and analyzed the subcellular localization of these fusion proteins. We show that the first 32 amino acids of Gi1α are sufficient to target GFP to caveolin-enriched domains of the plasma membrane in vivo, as demonstrated by co-fractionation and co-immunoprecipitation with caveolin-1. Interestingly, when dual acylation of this 32-amino acid domain was blocked by specific point mutations (G2A or C3S), the resulting GFP fusion proteins were localized to the cytoplasm and excluded from caveolin-rich regions. The myristoylated but nonpalmitoylated (C3S) chimera only partially partitioned into caveolin-containing fractions. However, both nonacylated GFP fusions (G2A and C3S) no longer co-immunoprecipitated with caveolin-1. Taken together, these results indicate that lipid modification of the NH2-terminal of Gi1α is essential for targeting to its correct destination and interaction with caveolin-1. Also, a caveolin-1 mutant lacking all three palmitoylation sites (C133S, C143S, and C156S) was unable to co-immunoprecipitate these dually acylated GFP-G-protein fusions. Thus, dual acylation of the NH2-terminal domain of Gi1α and palmitoylation of caveolin-1 are both required to stabilize and perhaps regulate this reciprocal interaction at the plasma membrane in vivo. Our results provide the first demonstration of a functional role for caveolin-1 palmitoylation in its interaction with signaling molecules

Microchemical and microscopic characterization of the pictorial quality of egg-tempera polyptych, late 14th century, Florence, Italy

This paper explores the added value of micro-chemical and microscopic approaches to gather scientific evidence that can technically explain the pictorial quality of an egg-tempera painting, and underpin assessments that otherwise would be based on naked eye observations only. Demonstration is here provided via the interdisciplinary investigation of the original technique used by Giovanni del Biondo in the polyptych Annunciation and Saints (1385 ca), Galleria dell'Accademia, Florence, Italy. The exquisite surface appearance makes this panel painting remarkable compared to artworks by coeval artists. Imaging techniques (UV, IR and IR false color), non-invasive single spot techniques (XRF and FORS spectrometry) and analytical investigations on eight selected micro-samples (ATR-FTIR, GC/MS and Py/GC–MS, ESEM-EDS) were combined to retrieve the palette and identify organic binding media and a superficial coating layer. Stratigraphic and micro-chemical data confirmed the use of a relatively simple egg-tempera technique applied on a ground made of gypsum mixed with animal glue, without complex stratigraphic superimposition of preparation and pictorial layers. Various pigments were identified, among which is the precious lapis lazuli. While Py/GC–MS highlight that the coating is made of dammar resin and honey mixed with animal glue, the results allow us to state that the painting was not intentionally varnished by Giovanni del Biondo. These outcomes shed a new light on the technical knowledge of this polyptych, and prove how challenging is the attempt to categorize egg-tempera recipes used by ancient painters at the turn of the 14th century

Molecular modeling studies, synthesis, configurational stability and biological activity of 8-chloro-2,3,5,6-tetrahydro-3,6-dimethyl-pyrrolo[1,2,3-de]-1,2,4-benzothiadiazine 1,1-dioxide

The potential therapeutic benefit of compounds able to activate AMPA receptors (AMPArs) has led to a search for new AMPAr positive modulators. Among them, 8-chloro-2,3,5,6-tetrahydro-3,6-dimethyl-pyrrolo[1,2,3-de]-1,2,4-benzothiadiazine 1,1-dioxide (1) has attracted particular attention, because it is one of the most active benzothiadiazine\u2013derived positive modulators of the AMPA receptor. It possesses two stereogenic centers, C3 and C6, thus it can exist as four stereoisomers. In this work, preliminary in silico studies suggested that 1 interacts stereoselectively with AMPArs. Single stereoisomers of 1 were prepared in order to evaluate their biological activity. However, studies regarding the configurational stability of the investigated compounds suggested a rapid epimerization at C3 in aqueous solvents, and we can expect the same reaction in vivo. Thus, electrophysiological experiments were performed on the two epimeric mixtures, (3 17,6R)- and (3 17,6S)- 8-chloro-2,3,5,6-tetrahydro-3,6-dimethyl-pyrrolo[1,2,3-de]-1,2,4-benzothiadiazine 1,1-dioxide, in order to evaluate their activities as positive allosteric modulators of AMPArs. The obtained data suggest that the (3 17,6S) epimeric mixture is the most active in positively modulating AMPArs, confirming in silico results

TIPOS DE PARTO E O PAPEL DO ENFERMEIRO

A expectativa das mulheres a respeito da escolha do parto tem relação com o conhecimento sobre o assunto e as informações que são passadas durante a gestação. O componente curricular Saúde da Mulher II, do Curso de graduação em Enfermagem, aborda assuntos relacionados ao ciclo grávido-puerperal. A gestação significa um momento especial para a mulher, e a decisão pela via de parto é influenciada por inúmeros fatores, como os riscos e os benefícios, possíveis complicações e repercussões futuras. Atualmente, por meio da humanização do parto, os profissionais da área da enfermagem possuem um papel importante durante o pré-natal e, consequentemente, com a gestante. Esse material foi elaborado com o objetivo de descrever os diversos tipos de parto, podendo ser de escolha da gestante ou por necessidade, suas vantagens e desvantagens e métodos aplicados. Os tipos de parto descritos foram: parto normal, parto normal sem dor, parto natural, parto na água, parto Laboyer, parto domiciliar, parto de cócoras e parto cesariana. Concluiu-se que o nascimento de um bebê é um momento muito especial na vida da mulher e da família, exatamente por isso é normal que existam várias dúvidas em torno da escolha ao realizar o parto, sendo fundamental que a mulher saiba sobre os diferentes tipos, as vantagens e desvantagens, informando e esclarecendo o que será melhor para o bem-estar dela e do seu filho. Encontra-se, então, o importante papel do profissional Enfermeiro, participando na realização do trabalho; a equipe multiprofissional tem uma importância enorme no momento em que a mãe e a família escolhem um tipo de parto para o nascimento de seu filho. Torna-se relevante o Enfermeiro entender quais os fatores que realmente influenciam as gestantes na decisão da via de parto, pois a partir desse conhecimento se pode direcionar a atenção e o cuidado à saúde da mãe e do bebê.Palavras-chave: Gestação. Tipos de parto. Enfermeiro. Cuidado mãe-bebê

Efficacy of Systemically Administered Retargeted Oncolytic Herpes Simplex Viruses—Clearance and Biodistribution in Naïve and HSV-Preimmune Mice

We investigated the anticancer efficacy, blood clearance, and tissue biodistribution of systemically administered retargeted oncolytic herpes simplex viruses (ReHVs) in HSV-naive and HSV-preimmunized (HSV-IMM) mice. Efficacy was tested against lung tumors formed upon intravenous administration of cancer cells, a model of metastatic disease, and against subcutaneous distant tumors. In naive mice, HER2- and hPSMA-retargeted viruses, both armed with mIL-12, were highly effective, even when administered to mice with well-developed tumors. Efficacy was higher for combination regimens with immune checkpoint inhibitors. A significant amount of infectious virus persisted in the blood for at least 1 h. Viral genomes, or fragments thereof, persisted in the blood and tissues for days. Remarkably, the only sites of viral replication were the lungs of tumor-positive mice and the subcutaneous tumors. No replication was detected in other tissues, strengthening the evidence of the high cancer specificity of ReHVs, a property that renders ReHVs suitable for systemic administration. In HSV-IMM mice, ReHVs administered at late times failed to exert anticancer efficacy, and the circulating virus was rapidly inactivated. Serum stability and in vivo whole blood stability assays highlighted neutralizing antibodies as the main factor in virus inactivation. Efforts to deplete mice of the neutralizing antibodies are ongoing

Design, stereoselective synthesis, configurational stability and biological activity of 7-chloro-9-(furan-3-yl)-2,3,3a,4-tetrahydro-1H-benzo[e]pyrrolo[2,1-c][1,2,4]thiadiazine 5,5-dioxide

Chiral 5-arylbenzothiadiazine derivatives have recently attracted particular attention because they exhibit an interesting pharmacological activity as AMPA receptor (AMPAr) positive modulators. However, investigations on their configurational stability suggest a rapid enantiomerization in physiological conditions. In order to enhance configurational stability, preserving AMPAr activity, we have designed the novel compound (R,S)-7-chloro-9-(furan-3-yl)-2,3,3a,4-tetrahydro-1H-benzo[e]pyrrolo[2,1-c][1,2,4]thiadiazine 5,5-dioxide bearing a pyrrolo moiety coupled with the 5-(furan-3-yl) substituent on benzothiadiazine core. A stereoselective synthesis was projected to obtain single enantiomer of the latter compound. Absolute configuration was assigned by X-ray crystal structure. Patch clamp experiments evaluating the activity of single enantiomers as AMPAr positive allosteric modulator showed that R stereoisomer is the active component. Molecular modeling studies were performed to explain biological results. An on-column stopped-flow bidimensional recycling HPLC procedure was applied to obtain on a large scale the active enantiomer with enantiomeric enrichment starting from the racemic mixture of the compound

Boneca terapêutica – ciclo grávido-puerperal

Apesar de a gravidez ser um fenômeno biologicamente normal, encerra profundas alterações e reestruturações em nível físico, em nível das estruturas internas e autopercepções, bem como das relações com os outros. Este estudo fez parte de uma atividade do Componente Curricular Saúde da Mulher III, do Curso de Enfermagem da Unoesc Joaçaba, com a qual se teve como objetivos formar profissionais que reflitam e sejam críticos no processo de ensino-aprendizagem, estimular o desenvolvimento da criatividade e de sua autonomia na aquisição de conhecimento, e utilizar o lúdico para despertar a curiosidade dos pacientes em entender o que é explicado. A confecção da boneca foi totalmente realizada pelos acadêmicos, mostrando que o enfermeiro pode e deve incluir o lúdico na sua rotina de trabalho. A técnica do brinquedo terapêutico busca minimizar o medo e a insegurança das mulheres, permitindo-lhes expressar seus anseios, sanar suas dúvidas e vivenciar de forma lúdica as situações que lhes causam angústia. A boneca tem por finalidade mostrar as fases da gravidez, as alterações no corpo da mulher e o crescimento do feto. Representa um período de intensas modificações físicas e psicológicas, transformando a vida social, familiar e profissional da gestante e de toda a família que está ao redor. Concluiu-se com a realização da atividade que os profissionais de saúde precisam olhar a mulher como um ser único, respeitando suas vontades e direitos, reconhecendo a mulher e o seu filho como peças fundamentais no evento do nascimento e compreendendo que não basta somente proporcionar à mulher um parto por via natural, se não forem considerados os sentimentos e desejos da parturiente e seus familiares, é de vital importância que se façam investimentos para estruturar melhor os hospitais e maternidades, remunerar melhor os profissionais e organizar espaços de aperfeiçoamento sobre técnicas e diretrizes para uma assistência humanizada; o processo gravídico puerperal demanda uma assistência digna e de qualidade que não se limita à expulsão ou extração de um feto do ventre da mulher, é um fenômeno que necessita a implementação de uma assistência verdadeiramente humanizada, com todos os profissionais da saúde, considerando os sentimentos e valores da mulher. A utilização da boneca nas atividades do enfermeiro contribui com esse processo de educação na comunidade.Palavras-chave: Enfermagem. Educação em saúde. Enfermagem obstétrica

Wilson Disease Protein ATP7B Utilizes Lysosomal Exocytosis to Maintain Copper Homeostasis

SummaryCopper is an essential yet toxic metal and its overload causes Wilson disease, a disorder due to mutations in copper transporter ATP7B. To remove excess copper into the bile, ATP7B traffics toward canalicular area of hepatocytes. However, the trafficking mechanisms of ATP7B remain elusive. Here, we show that, in response to elevated copper, ATP7B moves from the Golgi to lysosomes and imports metal into their lumen. ATP7B enables lysosomes to undergo exocytosis through the interaction with p62 subunit of dynactin that allows lysosome translocation toward the canalicular pole of hepatocytes. Activation of lysosomal exocytosis stimulates copper clearance from the hepatocytes and rescues the most frequent Wilson-disease-causing ATP7B mutant to the appropriate functional site. Our findings indicate that lysosomes serve as an important intermediate in ATP7B trafficking, whereas lysosomal exocytosis operates as an integral process in copper excretion and hence can be targeted for therapeutic approaches to combat Wilson disease

Perspectives on Open Science and scientific data sharing::an interdisciplinary workshop

Looking at Open Science and Open Data from a broad perspective. This is the idea behind "Scientific data sharing: an interdisciplinary workshop", an initiative designed to foster dialogue between scholars from different scientific domains which was organized by the Istituto Italiano di Antropologia in Anagni, Italy, 2-4 September 2013.We here report summaries of the presentations and discussions at the meeting. They deal with four sets of issues: (i) setting a common framework, a general discussion of open data principles, values and opportunities; (ii) insights into scientific practices, a view of the way in which the open data movement is developing in a variety of scientific domains (biology, psychology, epidemiology and archaeology); (iii) a case study of human genomics, which was a trail-blazer in data sharing, and which encapsulates the tension that can occur between large-scale data sharing and one of the boundaries of openness, the protection of individual data; (iv) open science and the public, based on a round table discussion about the public communication of science and the societal implications of open science. There were three proposals for the planning of further interdisciplinary initiatives on open science. Firstly, there is a need to integrate top-down initiatives by governments, institutions and journals with bottom-up approaches from the scientific community. Secondly, more should be done to popularize the societal benefits of open science, not only in providing the evidence needed by citizens to draw their own conclusions on scientific issues that are of concern to them, but also explaining the direct benefits of data sharing in areas such as the control of infectious disease. Finally, introducing arguments from social sciences and humanities in the educational dissemination of open data may help students become more profoundly engaged with Open Science and look at science from a broader perspective