SERS biosensor with plastic antibodies for detection of a cancer biomarker protein.

Surface-enhanced Raman scattering (SERS) is a powerful method for detecting breast cancer-specific biomarkers due to its extraordinary enhancement effects obtained by localized surface plasmon resonance (LSPR) in metallic nanostructures at hotspots. In this research, gold nanostars (AuNSs) were used as SERS probes to detect a cancer biomarker at very low concentrations. To this end, we combined molecularly imprinted polymers (MIPs) as a detection layer with SERS for the detection of the biomarker CA 15-3 in point-of-care (PoC) analysis. This required two main steps: (i) the deposition of MIPs on a gold electrode, followed by a second step (ii) antibody binding with AuNSs containing a suitable Raman reporter to enhance Raman signaling (SERS). The MPan sensor was prepared by electropolymerization of the monomer aniline in the presence of CA 15-3. The template molecule was then extracted from the polymer using sodium dodecyl sulfate (SDS). In parallel, a control material was prepared in the absence of the protein (NPan). Surface modification for the control was performed using electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The performance of the sensor was evaluated using the SERS technique, in which the MPan sensor is first incubated with the protein and then exposed to the SERS probe. Under optimized conditions, the device showed a linear response to CA 15-3 concentrations from 0.016 to 248.51 U mL-1 in a PBS buffer at pH 7.4 in 1000-fold diluted serum. Overall, this approach demonstrates the potential of SERS as an optical reader and opens a new avenue for biosensing applications.

Colorimetric Paper-Based Sensors against Cancer Biomarkers.

Cancer is a major cause of mortality and morbidity worldwide. Detection and quantification of cancer biomarkers plays a critical role in cancer early diagnosis, screening, and treatment. Clinicians, particularly in developing countries, deal with high costs and limited resources for diagnostic systems. Using low-cost substrates to develop sensor devices could be very helpful. The interest in paper-based sensors with colorimetric detection increased exponentially in the last decade as they meet the criteria for point-of-care (PoC) devices. Cellulose and different nanomaterials have been used as substrate and colorimetric probes, respectively, for these types of devices in their different designs as spot tests, lateral-flow assays, dipsticks, and microfluidic paper-based devices (µPADs), offering low-cost and disposable devices. However, the main challenge with these devices is their low sensitivity and lack of efficiency in performing quantitative measurements. This review includes an overview of the use of paper for the development of sensing devices focusing on colorimetric detection and their application to cancer biomarkers. We highlight recent works reporting the use of paper in the development of colorimetric sensors for cancer biomarkers, such as proteins, nucleic acids, and others. Finally, we discuss the main advantages of these types of devices and highlight their major pitfalls.

Recent Advances in the Selection of Cancer-Specific Aptamers for the Development of Biosensors.

An early diagnosis has the potential to greatly decrease cancer mortality. For that purpose, specific cancer biomarkers have been molecularly targeted by aptamer sequences to enable an accurate and rapid detection. Aptamer-based biosensors for cancer diagnostics are a promising alternative to those using antibodies, due to their high affinity and specificity to the target molecules and advantageous production. Synthetic nucleic acid aptamers are generated by in vitro Systematic Evolution of Ligands by Exponential enrichment (SELEX) methodologies that have been improved over the years to enhance the efficacy and shorten the selection process. Aptamers have been successfully applied in electrochemical, optical, photoelectrochemical and piezoelectrical-based detection strategies. These aptasensors comprise a sensitive, accurate and inexpensive option for cancer detection being used as point-of-care devices. This review highlights the recent advances in cancer biomarkers, achievements and optimizations made in aptamer selection, as well as the different aptasensors developed for the detection of several cancer biomarkers.

Diel biochemical and photosynthetic monitorization of Skeletonema costatum and Phaeodactylum tricornutum grown in outdoor pilot-scale flat panel photobioreactors.

Diatoms are currently considered valuable feedstocks for different biotechnological applications. To deepen the knowledge on the production of these microalgae, the diel pattern of batch growth, photosystem II performance, and accumulation of target metabolites of two commercially relevant diatoms, Phaeodactylum tricornutum and Skeletonema costatum, were followed outdoors in 100-L flat panel photobioreactors. S. costatum presented a higher light-to-biomass conversion resulting in higher growth than P. tricornutum. Both fluorescence data and principal component analysis pointed to temperature as a limiting factor for the growth of P. tricornutum. Higher protein and carbohydrate contents were found in P. tricornutum, whereas S. costatum fatty acids were characterized by a higher unsaturation degree. Higher productivities were found at 1 p.m. for protein, lipid, and ash in the case of S. costatum. Overall, S. costatum showed great potential for outdoor cultivation, revealing a broader temperature tolerance and increased biomass productivity than P. tricornutum.

Senses and meanings of medical professionalism for gynecology and obstetrics residents / Sentidos e significados de profissionalismo médico para residentes de ginecologia e obstetrícia

Abstract: Introduction: Professionalism is an essential condition for the doctor's performance. The specialty of Gynecology and Obstetrics (GO) has been marked by a growing demand for disciplinary proceedings. Faced with this reality, the American College of Gynecology and Obstetrics (ACOG) recommended recording destructive or hostile behaviors that affect patient safety and the quality of care, remediate them and educate the clinical staff of institutions about it. Despite its importance for the training of future professionals, the lack of consensus about its definition and measurement methods make it difficult to implement targeted actions aimed at this purpose. Objective: This study´s objective was to know the senses and meanings attributed by Gynecology and Obstetrics (GO) residents to medical professionalism. Method: An exploratory, qualitative study was carried out through focal groups with 21 GO residents from Fortaleza, Ceará, Brazil. The analysis of content in the thematic modality supported the data analysis and the interpretation was supported by Symbolic Interactionism. Results: The senses and meanings attributed to professionalism by the residents were expressed in four topics: set of conducts for the benefit of the patient; doctor-patient relationship and relationship with the work team; set of professional attributes; and moral and ethical exercise of the profession. Conclusion: Professionalism is understood as a set of characteristics that include the comprehensive training of the physician, based on skills, scientific knowledge, values, technical skills, clinical reasoning, enabling the exercise of Medicine with respect, morals and ethics.

Resumo: Introdução: Profissionalismo é condição essencial para a atuação do médico. A especialidade de ginecologia e obstetrícia (GO) tem sido marcada por uma demanda crescente de processos disciplinares. Diante dessa realidade, o Colégio Americano de Ginecologia e Obstetrícia (ACOG) recomendou registrar comportamentos destrutivos ou hostis que afetem a segurança do paciente e a qualidade do cuidado, remediá-los e educar o corpo clínico de instituições sobre isso. Apesar da importância do profissionalismo para a formação dos futuros profissionais, a ausência de consenso sobre a definição e os modos de mensuração desse aspecto dificulta a implementação de ações direcionadas voltadas a esse fim. Objetivo: Este estudo teve como objetivo conhecer os sentidos e significados atribuídos pelos residentes de GO ao profissionalismo médico. Método: Realizou-se estudo exploratório, qualitativo, por meio de grupos focais, com 21 residentes de GO de Fortaleza, no Ceará, no Brasil. A análise de conteúdo na modalidade temática respaldou a análise dos dados, e a interpretação amparou-se no interacionismo simbólico. Resultado: Os sentidos e significados atribuídos pelos residentes ao profissionalismo expressaram-se em quatro temáticas: conjunto de condutas em benefício do paciente; relação médico-paciente e com a equipe de trabalho; conjunto de atributos profissionais; e exercício moral e ético da profissão. Conclusão: O profissionalismo é entendido como um conjunto de características que incluem a formação integral do médico, a partir de competências, conhecimentos científicos, valores, habilidades técnicas e raciocínio clínico, possibilitando o exercício da medicina com respeito, moral e ética.

In situ monitoring of chlorophyll a fluorescence in Nannochloropsis oceanica cultures to assess photochemical changes and the onset of lipid accumulation during nitrogen deprivation.

In situ chlorophyll a fluorescence measurements were applied to monitor changes in the photochemical variables of Nannochloropsis oceanica cultures under nitrogen-deplete and nitrogen-replete (control) conditions. In addition, growth, lipid, fatty acid, and pigment contents were also followed. In the control culture, growth was promoted along with pigment content, electron transport rate (ETR), and polyunsaturated fatty acids, while total lipid content and fatty acid saturation level diminished. Under nitrogen-deplete conditions, the culture showed a higher de-epoxidation state of the xanthophyll cycle pigments. Fast transients revealed a poor processing efficiency for electron transfer beyond QA , which was in line with the low ETR due to nitrogen depletion. Lipid content and the de-epoxidation state were the first biochemical variables triggered by the change in nutrient status, which coincided with a 20% drop in the in situ effective quantum yield of PSII (ΔF'/Fm '), and a raise in the Vj measurements. A good correlation was found between the changes in ΔF'/Fm ' and lipid content (r = -0.96, p < 0.01). The results confirm the reliability and applicability of in situ fluorescence measurements to monitor lipid induction in N. oceanica.

Isolation and Characterization of Novel Chlorella Vulgaris Mutants With Low Chlorophyll and Improved Protein Contents for Food Applications.

Microalgae are widely used as food supplements due to their high protein content, essential fatty acids and amino acids as well as carotenoids. The addition of microalgal biomass to food products (e.g., baked confectioneries) is a common strategy to attract novel consumers. However, organoleptic factors such as color, taste and smell can be decisive for the acceptability of foods supplemented with microalgae. The aim of this work was to develop chlorophyll-deficient mutants of Chlorella vulgaris by chemically induced random mutagenesis to obtain biomass with different pigmentations for nutritional applications. Using this strategy, two C. vulgaris mutants with yellow (MT01) and white (MT02) color were successfully isolated, scaled up and characterized. The changes in color of MT01 and MT02 mutant strains were due to an 80 and 99% decrease in their chlorophyll contents, respectively, as compared to the original wild type (WT) strain. Under heterotrophic growth, MT01 showed a growth performance similar to that of the WT, reaching a concentration of 5.84 and 6.06 g L-1, respectively, whereas MT02 displayed slightly lower growth (4.59 g L-1). When grown under a light intensity of 100 µmol m-2 s-1, the pigment content in MT01 increased without compromising growth, while MT02 was not able to grow under this light intensity, a strong indication that it became light-sensitive. The yellow color of MT01 in the dark was mainly due to the presence of the xanthophyll lutein. On the other hand, phytoene was the only carotenoid detected in MT02, which is known to be colorless. Concomitantly, MT02 contained the highest protein content, reaching 48.7% of DW, a 60% increase as compared to the WT. MT01 exhibited a 30% increase when compared to that of the WT, reaching a protein content of 39.5% of DW. Taken together, the results strongly suggest that the partial abrogation of pigment biosynthesis is a factor that might promote higher protein contents in this species. Moreover, because of their higher protein and lower chlorophyll contents, the MT01 and MT02 strains are likely candidates to be feedstocks for the development of novel, innovative food supplements and foods.

O profissionalismo e suas formas de avaliação em estudantes de Medicina: uma revisão integrativa / El profesionalismo y sus formas de evaluación en estudiantes de Medicina: una revisión integradora / Professionalism and its forms of assessment in medical students: An integrative review

Com o objetivo de sistematizar informações sobre avaliação de profissionalismo em estudantes de Medicina, foi realizada em maio de 2018, revisão integrativa nas bases EBSCO, LILACS, MEDLINE, via PubMed no período de 2013 a 2018, sendo encontrados 12 artigos e três eixos temáticos: profissionalismo: constructo multidimensional e indefinido; ensino de profissionalismo médico; avaliando o profissionalismo: estratégias múltiplas de avaliação. A avaliação por meio de domínios como altruísmo, responsabilidade, cuidado, trabalho em equipe, é frequente, podendo ser utilizado cenários ou pacientes simulados; questionários online ou aplicativos. Ressalta-se a importância da modelagem no ensino de profissionalismo e o uso do feedback associado a outros métodos de avaliação. Não existe conceito único de profissionalismo. Percebe-se a necessidade de associação de métodos para uma melhor avaliação.(AU)

Con el objetivo de sistematizar informaciones sobre evaluación de profesionalismo en estudiantes de Medicina, en mayo de 2018 se realizó una revisión integradora en las bases EBSCO, LILACS, MEDLINE, vía PubMed en el período de 2013 a 2018, habiéndose encontrado 12 artículos y tres ejes temáticos: profesionalismo: construcción multidimensional e indefinida; enseñanza de profesionalismo médico; evaluación del profesionalismo: estrategias múltiples de evaluación. La evaluación por medio de dominios tales como altruismo, responsabilidad, cuidado y trabajo en equipo es frecuente, pudiendo utilizarse escenarios o pacientes simulados; cuestionarios online o aplicaciones. Se subraya la importancia del modelado en la enseñanza del profesionalismo y el uso del feedback asociado a otros métodos de evaluación. No hay concepto único de profesionalismo. Se percibe la necesidad de asociación de métodos para una mejor evaluación.(AU)

With the objective of systematizing information on the assessment of professionalism among medical students, an integrative review was carried out in May 2018 in the databases EBSCOhost, LILACS and MEDLINE, via PubMed, from 2013 to 2018. Overall, 12 articles and three thematic axes were found: Professionalism: a multidimensional and undefined construct; The teaching of medical professionalism; and Assessing professionalism: multiple assessment strategies. Assessment through domains such as altruism, responsibility, care and teamwork is frequent, and simulated scenarios, standardized patients, online questionnaires or applications may be used. The importance of modeling in the teaching of professionalism is emphasized, as well as the use of feedback associated with other assessment methods. We concluded that there is not one single concept of professionalism and noticed that better assessments are produced when different methods are associated.(AU)

Dual biorecognition by combining molecularly-imprinted polymer and antibody in SERS detection. Application to carcinoembryonic antigen.

This work reports the innovative combination of a molecularly-imprinted polymer (MIP) and a natural antibody for the accurate surface-enhanced Raman spectroscopy (SERS) detection of carcinoembryonic antigen (CEA). The MIP material acted as a pre-concentration scheme for the target protein, while the natural antibody was responsible to signal the presence of CEA on the MIP platform. Gold-based screen-printed electrodes were used as substrate and gallic acid (GA) was used herein for the first time in the assembly of a MIP film, by electropolymerization, in the presence of CEA. This layer was further covered by a second ultra-thin film of electropolymerized benzoic acid (BA), to avoid non-specific binding. The rebinding features of the MIP film were evaluated by electrochemical impedance spectroscopy (EIS) and a linear response was observed from 1 to 1000 ng/mL. For a sensitive SERS detection, the MIP film was first incubated in sample containing CEA and next incubated in SERS tag. For the SERS tag, gold nanostars (AuNSs) were employed as metal support, coupled to 4-aminothiophenol (4-ATP) as Raman reporter and to a natural antibody for CEA as recognition element. The overall system showed a sensitive response down to 1.0 ng/mL, which was different from the blank signal. Overall, the innovative approach presented herein combines the advantages of using two different targeting elements for CEA. The costs and time of MIP production were substantially low due to selection of electropolymerization approach and the proposal described herein may be extended to other target molecules.

Professionalism in the Training of Medical Specialists: an Integrative Literature Review / Profissionalismo na Formação do Especialista Médico: Revisão Integrativa da Literatura

ABSTRACT Introduction The construct professionalism is increasingly being recognized as an important component of medical education; fundamental to the physician's role in society Objective: To group in a concise and systemized way the information available in scientific productions on the concept of medical professionalism and its applications in medical residency programs. Methods A total of 85 articles were found in 2018, of which 13 were excluded for duplication using the Mendeley platform. Of the 72 remaining articles, 35 were excluded for not answering the research question. Of the 37 articles selected for reading of the abstract, eight were not available, leaving 29 articles. After reading all articles for the final selection, 11 articles were excluded. Six papers did not fulfill the goals of the research, and five were opinion articles. Results The production included 18 articles, from which analysis emerged three thematic categories: (a) professionalism: multidimensional construct; (b) teaching of professionalism: role modeling and the curriculum; (c) evaluation of professionalism: multiple strategies in the curriculum. The most frequently cited domains were: altruism, responsibility, care, teamwork, self-control, ethical principles and clinic excellence. Ten articles highlighted the importance of role modeling by preceptors and members of the health team in the Medical Residency Program for teaching professionalism. Regarding the most commonly used strategies for the selection, feedback practices and the OSCE were highlighted in scenarios to evaluate the domains of professionalism. Conclusion Professionalism is a dynamic construct that is worked on in the form of teaching and assessment in Medical Residency Programs. There is no consensus on the professionalism concept, and thus, multiple strategies are used for the teaching, among which role modeling can be highlighted. The teaching of medical professionalism, as a construct, in the training of specialists can reduce lapses and foster assistance based on respect for the autonomy of people and social responsibility.

RESUMO Introdução O constructo profissionalismo vem, cada vez mais, sendo reconhecido como importante componente da educação médica, sendo fundamental na atuação do médico na sociedade. Objetivo Reunir de forma sucinta e sistematizada as informações disponíveis nas produções científicas sobre o conceito de profissionalismo médico e sua aplicação nos programas de residência médica. Métodos Foram encontrados 85 artigos em maio de 2018, dos quais 13 foram excluídos por duplicidade com o uso da plataforma Mendeley. Dos 72 artigos restantes, 35 foram excluídos por não responderem à pergunta de pesquisa. Dos 37 artigos selecionados pela leitura dos resumos, 8 não estavam disponíveis, restando 29 artigos. Após leitura integral de todos os artigos para a seleção definitiva, foram eliminadas 11 publicações. Destas, 6 artigos não responderam aos objetivos da pesquisa e 5 eram artigos de opinião. Resultados A amostra incluiu 18 artigos, de cuja análise emergiram três categorias temáticas: (a) profissionalismo: constructo multidimensional; (b) ensino de profissionalismo: papel da modelagem e do currículo; (c) avaliação de profissionalismo: estratégias múltiplas no currículo. Os domínios mais citados foram: altruísmo, responsabilidade, cuidado, trabalho em equipe, autocontrole, princípios éticos e excelência clínica. Dez artigos destacaram a importância da modelagem por preceptores e membros da equipe de saúde no programa de residência médica para o ensino de profissionalismo. Em relação às estratégias mais utilizadas para avaliação, destacaram-se as práticas de feedback e Osce em cenários para avaliar os domínios de profissionalismo. Conclusão Profissionalismo é um constructo dinâmico que é trabalhado na forma de domínios no ensino e avaliação nos Programas de Residência Médica. Não há consenso sobre o conceito de profissionalismo, sendo utilizadas estratégias múltiplas para o ensino, com destaque para a modelagem. O ensino do profissionalismo médico, enquanto constructo, na formação de especialistas pode reduzir lapsos e fomentar uma assistência pautada no respeito à autonomia das pessoas e na responsabilização social.

GST transcriptional changes induced by a toxic Microcystis aeruginosa strain in two bivalve species during exposure and recovery phases.

Previous studies have demonstrated the modulation of glutathione transferases (GSTs) induced by microcystin (MC) alone or in combination with other cyanobacterial secondary metabolites in bivalves. However, interspecies information about which and how GST isoforms are affected by these secondary metabolites is still scarce, especially considering the dynamic process involving their uptake and elimination routes. In this context, the role of GSTs gene expression changes in response to a toxic Microcystis aeruginosa extract were examined for Mytilus galloprovincialis and Ruditapes philippinarum during exposure and recovery phases. The expression levels of sigma 1, sigma 2, pi and mu-class GST genes were analyzed in the hepatopancreas of both bivalve species during cyanobacteria extract exposure (24 h) and post-exposure (24 and 72 h). Only a significant induction of sigma 1-class GST expression was observed for R. philippinarum upon 24-hour exposure of both bivalve species to Microcystis extract. During the recovery phase, GST transcriptional changes for M. galloprovincialis were characterized by an early induction (24 h) of sigma 1 and sigma 2 transcripts. On the other hand, GST transcriptional changes for R. philippinarum during post-exposure phase were characterized by an early induction (24 h) of sigma 1 and mu transcripts and a later induction (72 h) of the four analyzed GST transcripts. Such differences reflect variable GST response mechanisms to cope with MC-producing cyanobacterial blooms exposure between these two bivalve species, revealing a higher sensitivity of R. philippinarum to Microcystis-induced stress than M. galloprovincialis. The results also suggest a much higher level of activity of the GST detoxification system during the recovery phase compared to the period of the stress exposure for both bivalve species.

Canine leishmaniasis: Genome-wide analysis and antibody response to Lutzomyia longipalpis saliva.

The anti-inflammatory properties of sand fly saliva favor the establishment of the Leishmania infantum infection. In contrast, an antibody response against Lutzomyia longipalpis saliva is often associated with a protective cell-mediated response against canine visceral leishmaniasis. Genetic studies may demonstrate to what extent the ability to secrete anti-saliva antibodies depends on genetic or environmental factors. However, the genetic basis of canine antibody response against sand fly saliva has not been assessed. The aim of this study was to identify chromosomal regions associated with the anti-Lu. longipalpis salivary IgG response in 189 dogs resident in endemic areas in order to provide information for prophylactic strategies. Dogs were classified into five groups based on serological and parasitological diagnosis and clinical evaluation. Anti-salivary gland homogenate (SGH) IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). Genomic DNA was isolated from blood samples and genotyped using a SNP chip with 173,662 single nucleotide polymorphism (SNP) markers. The following linear regression model was fitted: IgG level = mean + origin + sex + age + use of a repellent collar, and the residuals were assumed as pseudo-phenotypes for the association test between phenotypes and genotypes (GWA). A component of variance model that takes into account polygenic and sample structure effects (EMMAX) was employed for GWA. Phenotypic findings indicated that anti-SGH IgG levels remained higher in exposed and subclinically infected dogs than in severely diseased dogs even in regression model residuals. Five associated markers were identified on chromosomes 2, 20 and 31. The mapped genes included CD180 (RP105) and MITF related to the rapid activation of B lymphocytes and differentiation into antibody-secreting plasma cells. The findings pointed to chromosomal segments useful for functional confirmation studies and a search for adjuvant molecules of the anti-saliva response.

The Goat (Capra hircus) Mammary Gland Mitochondrial Proteome: A Study on the Effect of Weight Loss Using Blue-Native PAGE and Two-Dimensional Gel Electrophoresis.

Seasonal weight loss (SWL) is the most important limitation to animal production in the Tropical and Mediterranean regions, conditioning producer's incomes and the nutritional status of rural communities. It is of importance to produce strategies to oppose adverse effects of SWL. Breeds that have evolved in harsh climates have acquired tolerance to SWL through selection. Most of the factors determining such ability are related to changes in biochemical pathways as affected by SWL. In this study, a gel based proteomics strategy (BN: Blue-Native Page and 2DE: Two-dimensional gel electrophoresis) was used to characterize the mitochondrial proteome of the secretory tissue of the goat mammary gland. In addition, we have conducted an investigation of the effects of weight loss in two goat breeds with different levels of adaptation to nutritional stress: Majorera (tolerant) and Palmera (susceptible). The study used Majorera and Palmera dairy goats, divided in 4 sets, 2 for each breed: underfed group fed on wheat straw (restricted diet, so their body weight would be 15-20% reduced by the end of experiment), and a control group fed with an energy-balanced diet. At the end of the experimental period (22 days), mammary gland biopsies were obtained for all experimental groups. The proteomic analysis of the mitochondria enabled the resolution of a total of 277 proteins, and 148 (53%) were identified by MALDI-TOF/TOF mass spectrometry. Some of the proteins were identified as subunits of the glutamate dehydrogenase complex and the respiratory complexes I, II, IV, V from mitochondria, as well as numerous other proteins with functions in: metabolism, development, localization, cellular organization and biogenesis, biological regulation, response to stimulus, among others, that were mapped in both BN and 2DE gels. The comparative proteomics analysis enabled the identification of several proteins: NADH-ubiquinone oxidoreductase 75 kDa subunit and lamin B1 mitochondrial (up-regulated in the Palmera breed), Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2 (up-regulated in the Majorera breed) and cytochrome b-c1 complex subunit 1, mitochondrial and Chain D, Bovine F1-C8 Sub-Complex Of Atp Synthase (down-regulated in the Majorera breed) as a consequence of weight loss.

The prion protein selectively binds to and modulates the content of purinergic receptor P2X4R.

The GPI-anchored prion protein (PrP(C)) is involved in neurodegeneration, either through misfolding in the Transmissible Spongiform Encephalopathies (TSE), or as a mediator of the neurotoxicity of peptide oligomers in Alzheimer's Disease. PrP(C) has been attributed pleiotropic functions, and appears to scaffold a variety of cell surface signaling modules, for example through its binding to several neurotransmitter receptors. Here we used transfected HEK293 cells to test for an interaction of PrP(C) with purinergic receptor P2X4R. The prion protein bound P2X4R in both overlay and co-immunoprecipitation assays, and co-localized mostly intracellularly, but occasionaly at the cell surface in confocal micrographs. Functional PrP(C):P2X4R interaction was tested by the uptake of a P2X4R-permeant compound, and by modulation of intracellular calcium. Unexpectedly, however, this interaction was traced to a selective effect of PrP(C) upon the content of co-transfected P2X4R. The results suggest a role of PrP(C) in proteostasis, dysfunctions of which may be involved in the pathogenesis of neurodegenerative diseases such as TSE and Alzheimer's Disease.

Glutathione Transferases Responses Induced by Microcystin-LR in the Gills and Hepatopancreas of the Clam Venerupis philippinarum.

A multi-method approach was employed to compare the responses of Glutatione Transferases (GSTs) in the gills and hepatopancreas of Venerupis philippinarum to microcystins (MCs) toxicity. In this way, using the cytosolic fraction, the enzymatic activity of GSTs, superoxide dismutase (SOD), serine/threonine protein phosphatases (PPP2) along with the gene expression levels of four GST isoforms (pi, mu, sigma1, sigma2) were investigated in both organs of the clams exposed for 24 h to 10, 50 and 100 µg L(-1) of MC-LR. Cytosolic GSTs (cGSTs) from both organs of the high dose exposed clams were purified by glutathione-agarose affinity chromatography, characterized kinetically and the changes in the expression of cGSTs of the gills identified using a proteomic approach. MC-LR caused an increase in GST enzyme activity, involved in conjugation reactions, in both gills and hepatopancreas (100 µg L(-1) exposure). SOD activity, an indicator of oxidative stress, showed significantly elevated levels in the hepatopancreas only (50 and 100 µg L(-1) exposure). No significant changes were found in PPP2 activity, the main target of MCs, for both organs. Transcription responses revealed an up-regulation of sigma2 in the hepatopancreas at the high dose, but no significant changes were detected in the gills. Kinetic analysis evidenced differences between gills of exposed and non-exposed extracts. Using proteomics, qualitative and quantitative differences were found between the basal and inducible cGSTs. Overall, results suggest a distinct role of GST system in counteracting MCs toxicity between the gills and the hepatopancreas of V. philippinarum, revealing different roles between GST isoforms within and among both organs.

Proteomic analysis of anatoxin-a acute toxicity in zebrafish reveals gender specific responses and additional mechanisms of cell stress.

Anatoxin-a is a potent neurotoxin produced by several genera of cyanobacteria. Deaths of wild and domestic animals due to anatoxin-a exposure have been reported following a toxic response that is driven by the inhibition of the acetylcholine receptors at neuromuscular junctions. The consequent neuron depolarization results in an overstimulation of the muscle cells. In order to unravel further molecular events implicated in the toxicity of anatoxin-a, a proteomic investigation was conducted. Applying two-dimensional gel electrophoresis (2DE) and MALDI-TOF mass spectrometry, we report early proteome changes in brain and muscle of zebrafish (Danio rerio) caused by acute exposure to anatoxin-a. In this regard, the test group of male and female zebrafish received an intraperitoneal (i.p.) injection of an anatoxin-a dose of 0.8µgg(-1) of fish body weight (bw) in phosphate buffered saline solution (PBS), while the control received an i.p. injection of PBS only. Five minutes after i.p. injection, brain and muscle tissues were collected, processed and analyzed with 2DE. Qualitative and quantitative analyzes of protein abundance allowed the detection of differences in the proteome of control and exposed fish groups, and between male and female fish (gender specific responses). The altered proteins play functions in carbohydrate metabolism and energy production, ATP synthesis, cell structure maintenance, cellular transport, protein folding, stress response, detoxification and protease inhibition. These changes provide additional insights relative to the toxicity of anatoxin-a in fish. Taking into account the short time of response considered (5min of response to the toxin), the changes in the proteome observed in this work are more likely to derive from fast occurring reactions in the cells. These could occur by protein activity regulation through degradation (proteolysis) and/or post-translational modifications, than from a differential regulation of gene expression, which may require more time for proteins to be synthesized and to produce changes at the proteomic level.

Transcriptional responses of glutathione transferase genes in Ruditapes philippinarum exposed to microcystin-LR.

Glutathione Transferases (GSTs) are phase II detoxification enzymes known to be involved in the molecular response against microcystins (MCs) induced toxicity. However, the individual role of the several GST isoforms in the MC detoxification process is still unknown. In this study, the time-dependent changes on gene expression of several GST isoforms (pi, mu, sigma 1, sigma 2) in parallel with enzymatic activity of total GST were investigated in gills and hepatopancreas of the bivalve Ruditapes philippinarum exposed to pure MC-LR (10 and 100 µg/L). No significant changes in GST enzyme activities were found on both organs. In contrast, MC-LR affected the transcriptional activities of these detoxification enzymes both in gills and hepatopancreas. GST transcriptional changes in gills promoted by MC-LR were characterized by an early (12 h) induction of mu and sigma 1 transcripts. On the other hand, the GST transcriptional changes in hepatopancreas were characterized by a later induction (48 h) of mu transcript, but also by an early inhibition (6 h) of the four transcripts. The different transcription patterns obtained for the tested GST isoforms in this study highlight the potential divergent physiological roles played by these isoenzymes during the detoxification of MC-LR.

Escaneamento de modelos para a confecção de setup virtual / Scanning models for virtual setup

O setup ortodôntico é uma excelente ferramenta para planejar o tratamento de um paciente. Quando bem executado, auxilia no planejamento do tratamento e da mecânica, e ajuda o paciente a entender as modificações dentárias propostas. O método mais frequentemente utilizado para a confecção do setup é a partir de modelos de gesso; entretanto, a possibilidade de obter modelos virtuais das arcadas dentárias permitiu o desenvolvimento de um novo método de confecção de setup: o método virtual. / Objetivo / Demonstrar, passo a passo, as etapas para confecção de um setup virtual a partir de modelos virtuais obtidos após o escaneamento de modelos de gesso. / Métodos / Um modelo de gesso de um paciente com indicação de extração de quatro pré-molares foi escaneado com aparelho 3D Shape R700. O arquivo foi importado pelo software OrthoAnalyzer 3D, e foi confeccionado um setup virtual para o caso em questão. / Conclusão / O presente artigo apresenta uma proposta viável e exequível de setup virtual realizado com o software OrthoAnalyzer 3D, após o escaneamento dos modelos de gesso com o aparelho 3D Shape R700.

Fisioterapia no pós-operatório de fratura proximal do fêmur em idosos: revisão da literatura / Physical therapy in the post-operative of proximal femur fracture in elderly: Literature review

As fraturas proximais do fêmur em idosos representam um sério problema de saúde publica. O tratamento cirúrgico dessa fratura serve para reduzir as morbidades, juntamente com a fisioterapia pós-operatória. O objetivo foi realizar uma revisão sistemática sobre protocolos de fisioterapia no pós-operatório de fraturas proximais de fêmur em idosos. Foram selecionados ensaios clínicos controlados e randomizados, em idosos, dos últimos 10 anos, nos idiomas português e inglês. Foram encontrados 14 artigos na literatura. A fisioterapia tem um papel importante na recuperação funcional desses idosos. Nível de Evidência I, Revisão Sistemática de ECRC (Estudos clínicos randomizados e controlados).

The proximal femoral fractures in the elderly is a serious public health problem. Surgical treatment of this fracture are used to reduce morbidity, together with postoperative physical therapy. The objective was to conduct a systematic review of physical therapy protocols in postoperative for fractures of the proximal femur in elderly. We selected randomized controlled trials in elderly in the past 10 years, in Portuguese and English. There were 14 articles in the literature. Physical therapy has an important role in functional recovery of the elderly. Level of Evidence I, Systematic Review RCTs (Study results were homogenous).

Physical therapy in the postoperative of proximal femur fracture in elderly. Literature review.

The proximal femoral fracture in the elderly is a serious public health problem. Surgical treatment of this fracture is used to reduce morbidity, together with postoperative physical therapy. The objective was to conduct a systematic review of physical therapy protocols in postoperative for fractures of the proximal femur in elderly. We selected randomized controlled trials in elderly in the past 10 years, in Portuguese and English. There were 14 articles in the literature. Physical therapy has an important role in functional recovery of the elderly. Level of Evidence I, Systematic Review RCTs (Study results were homogenous).