Diffusion dans un hydrogel : applications aux biocapteurs et optimisation de la technique de spectroscopie par corrélation de fluorescence (FCS)

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The difficulty of measuring culture and diversity. Comparison of three international models of cultural measurement: FCS-2009, ESSnet-2012 and CAB-2015

The growing interest in quantifying the cultural and creative industries, visualize the economic contribution of activities related to culture demands first of all the construction of internationally comparable analysis frameworks. Currently there are three major bodies which address this issue and whose comparative study is the focus of this article: the UNESCO Framework for Cultural Statistics (FCS-2009), the European Framework for Cultural Statistics (ESSnet-Culture 2012) and the methodological resource of the “Convenio Andrés Bello” group for working with the Satellite Accounts on Culture in Ibero-America (CAB-2015). Cultural sector measurements provide the information necessary for correct planning of cultural policies which in turn leads to sustaining industries and promoting cultural diversity. The text identifies the existing differences in the three models and three levels of analysis, the sectors, the cultural activities and the criteria that each one uses in order to determine the distribution of the activities by sector. The end result leaves the impossibility of comparing cultural statistics of countries that implement different frameworks.

Aferição das dificuldades/competências na área da Endodontia percepcionadas pelos alunos do 4º e 5º anos da FCS-UFP

Introdução: Com uma consciencialização cada vez maior das populações para a preservação dos dentes naturais, a Endodontia tem assumido uma importância crescente, tendo por objectivo principal a manutenção de dentes funcionais, sem prejudicar a saúde dos pacientes. Estes devem beneficiar de um tratamento segundo o “standard of care”, proporcionado por profissionais competentes. Vários estudos demonstram que a Endodontia é considerada uma área difícil e stressante para os estudantes, exibindo um sentimento de menor confiança na prática clínica, sobretudo no que diz respeito a procedimentos mais complexos, nomeadamente, no tratamento de dentes posteriores. Em 2010, na tentativa de homogeneizar as competências adquiridas pelos Médicos Dentistas (MD), a Associação para a Educação Dentária Europeia (AEDE) definiu critérios para a sua formação pré-graduada, indicando que o MD recém-graduado deve ter adquirido competências e capacidades técnicas que lhe permita começar a sua prática clínica de forma independente. Objectivos: Analisar os conhecimentos adquiridos e as dificuldades sentidas na área da Endodontia pelos alunos do 4º e 5º anos do Mestrado Integrado em Medicina Dentária da Faculdade de Ciências de Saúde (FCS) da Universidade Fernando Pessoa (UFP), fazendo, igualmente, uma abordagem ao ensino ministrado na área da Endodontia face aos resultados obtidos. Materiais e Métodos: Este trabalho está dividido em duas partes: revisão bibliográfica e investigação científica. A revisão bibliográfica do tema engloba o ensino graduado em Medicina Dentária, o uso de isolamento absoluto (IA) na prática clínica de Endodontia, a instrumentação manual versus rotatória, a influência no sucesso do tratamento endodôntico (TE) da sua execução por estudantes do 4º e 5º ano, as Guidelines para o TE e o ensino de Endodontia na UFP. A pesquisa bibliográfica foi efectuada através da base de dados PubMed, tendo sido utilizadas, em diferentes combinações, as seguintes palavras-chave: ”Endodontics”, “teaching”; “pre-clinical”; “undergraduate”; “Clinical”; “treatment” e “Europe”. O trabalho de investigação consistiu na elaboração, aprovação pela Comissão de Ética da UFP e posterior aplicação de um questionário destinado aos alunos do 4º e 5º ano do Mestrado Integrado em Medicina Dentária da FCS-UFP. A população estimada de alunos do 4º ano foi de 190 alunos e a de 5º ano de 150 alunos, o que perfaz um total de 340 alunos. Dos inquéritos respondidos, apenas foram considerados 338, uma vez que dois foram anulados pois continham respostas inválidas. Resultados: A “condição socioeconómica do paciente” foi considerada a causa menos relevante para avaliar um caso endodôntico, tendo sido apontada por 34,3% dos alunos. Assinala-se que 92% dos alunos de 4º ano e 93,3% dos alunos de 5º ano consideram que os molares superiores são os dentes mais difíceis de tratar, sendo a visibilidade uma das principais razões para esta opinião. O grau de dificuldade para a colocação do IA é definido como “Elevado” para a maioria dos alunos de 4º ano. A maior parte dos alunos de 5º ano considera que o grau de dificuldade para a “Determinação do tipo de reconstrução/prótese fixa mais indicada” é “Elevado”. O passo do TE onde os alunos do 4º ano se sentem mais confiantes é no “Diagnóstico de cárie”, sendo que os alunos do 5º ano se sentem mais confiantes ao realizar o “TE em dentes com 1 ou 2 canais”. O “Conteúdo leccionado nas aulas teóricas” foi considerado o principal aspecto positivo do ensino endodôntico. Por outro lado, o “Número de actos clínicos realizados” foi considerado o principal aspecto negativo, tanto para alunos de 4º como de 5º ano. Em ambos os anos, os principais pontos que os alunos acham que devem ser melhorados são o “Número de pacientes nas aulas clínicas” e a “Aprendizagem da técnica de instrumentação mecanizada/obturação termoplástica”. As principais preocupações referidas, tanto por alunos de 4º como de 5º anos, foram a “Insegurança na prática clínica” e a “Dificuldade em encontrar ofertas profissionais”. Relativamente à qualidade dos seus TE existe um número significativo de alunos do 5º ano que já se auto-avaliam como “Bons” (33,6%), por comparação com os do 4º ano (21,2%), sendo a auto-avaliação de “Razoável” dominante em ambos os anos de formação. Conclusões: Conclui-se que na disciplina de Endodontia na FCS-UFP, são seguidas as Guidelines da European Society of Endodontology (ESE), sendo que estas indicam o protocolo mais correcto a seguir durante o Tratamento Endodôntico Não-Cirúrgico (TENC). Contudo, o reduzido número de actos clínicos e a consequente falta de prática faz com que os alunos se sintam pouco confiantes ao iniciar a sua actividade profissional.

Measuring The Hydrodynamic Radius Of Quantum Dots By Fluorescence Correlation Spectroscopy.

Fluorescence Correlation Spectroscopy (FCS) is an optical technique that allows the measurement of the diffusion coefficient of molecules in a diluted sample. From the diffusion coefficient it is possible to calculate the hydrodynamic radius of the molecules. For colloidal quantum dots (QDs) the hydrodynamic radius is valuable information to study interactions with other molecules or other QDs. In this chapter we describe the main aspects of the technique and how to use it to calculate the hydrodynamic radius of quantum dots (QDs).

Measurement Of The Hydrodynamic Radius Of Quantum Dots By Fluorescence Correlation Spectroscopy Excluding Blinking.

One of the most important properties of quantum dots (QDs) is their size. Their size will determine optical properties and in a colloidal medium their range of interaction. The most common techniques used to measure QD size are transmission electron microscopy (TEM) and X-ray diffraction. However, these techniques demand the sample to be dried and under a vacuum. This way any hydrodynamic information is excluded and the preparation process may alter even the size of the QDs. Fluorescence correlation spectroscopy (FCS) is an optical technique with single molecule sensitivity capable of extracting the hydrodynamic radius (HR) of the QDs. The main drawback of FCS is the blinking phenomenon that alters the correlation function implicating in a QD apparent size smaller than it really is. In this work, we developed a method to exclude blinking of the FCS and measured the HR of colloidal QDs. We compared our results with TEM images, and the HR obtained by FCS is higher than the radius measured by TEM. We attribute this difference to the cap layer of the QD that cannot be seen in the TEM images.

Biofilm And Saliva Affect The Biomechanical Behavior Of Dental Implants.

Friction coefficient (FC) was quantified between titanium-titanium (Ti-Ti) and titanium-zirconia (Ti-Zr), materials commonly used as abutment and implants, in the presence of a multispecies biofilm (Bf) or salivary pellicle (Pel). Furthermore, FC was used as a parameter to evaluate the biomechanical behavior of a single implant-supported restoration. Interface between Ti-Ti and Ti-Zr without Pel or Bf was used as control (Ctrl). FC was recorded using tribometer and analyzed by two-way Anova and Tukey test (p<0.05). Data were transposed to a finite element model of a dental implant-supported restoration. Models were obtained varying abutment material (Ti and Zr) and FCs recorded (Bf, Pel, and Ctrl). Maximum and shear stress were calculated for bone and equivalent von Misses for prosthetic components. Data were analyzed using two-way ANOVA (p<0.05) and percentage of contribution for each condition (material and FC) was calculated. FC significant differences were observed between Ti-Ti and Ti-Zr for Ctrl and Bf groups, with lower values for Ti-Zr (p<0.05). Within each material group, Ti-Ti differed between all treatments (p<0.05) and for Ti-Zr, only Pel showed higher values compared with Ctrl and Bf (p<0.05). FC contributed to 89.83% (p<0.05) of the stress in the screw, decreasing the stress when the FC was lower. FC resulted in an increase of 59.78% of maximum stress in cortical bone (p=0.05). It can be concluded that the shift of the FC due to the presence of Pel or Bf is able to jeopardize the biomechanical behavior of a single implant-supported restoration.

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Melatonin (MEL) acts as a powerful scavenger of free radicals and direct gonadal responses to melatonin have been reported in the literature. Few studies, however, have evaluated the effect of MEL during in vitro maturation (IVM) on bovine embryos. This study tested the addition of MEL to maturation medium (MM) with no gonadotropins on nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos. Cumulus-oocyte complexes were aspirated from abattoir ovaries and cultured in MM (TCM-199 medium supplemented with 10% fetal calf serum - FCS) at 39ºC and 5% CO2 in air. After 24 hours of culture in MM with 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH; 10-9 M MEL) or 10-9 M MEL, 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH, the oocytes were stained with Hoechst 33342 to evaluate nuclear maturation rate. After in vitro fertilization and embryo culture, development rates were evaluated and the blastocysts were assessed for DNA damage by Comet assay. There was no effect of melatonin added to the MM, alone or in combination with gonadotropins, on nuclear maturation, cleavage and blastocyst rates. These rates ranged between 88% to 90%, 85% to 88% and 42% to 46%, respectively. The extent of DNA damage in embryos was also not affected by MEL supplementation during IVM. The addition of 10-9 M MEL to the MM failed to improve nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos, but was able to properly substitute for gonadotropins during IVM.

Dietary salt restriction increases plasma lipoprotein and inflammatory marker concentrations in hypertensive patients

Background: Dietary salt restriction has been reported to adversely modify the plasma lipoprotein profile in hypertensive and in normotensive subjects. We investigated the effects of the low sodium intake (LSI) on the plasma lipoprotein profile and on inflammation and thrombosis biomarkers during the fasting and postprandial periods. Methods: Non-obese, non-treated hypertensive adults (n=41) were fed strictly controlled diets. An initial week on a control diet (CID, Na=160 mmol/day) was followed by 3 weeks on LSI (Na=60mmol/day). At admission and on the last day of each period, the 24-h ambulatory blood pressure was monitored and blood was drawn after an overnight fasting period and after a fat-rich test meal. Results: The dietary adherence was confirmed by 24-h urinary sodium excretion. Fasting triglyceride (TG), chylomicron-cholesterol, hsC-reactive protein (CRP), tumor necrosis factor-a (TNF-alpha). interleukin-6 (IL-6) concentrations, renin activity, aldosterone, insulin, and homeostasis model assessment insulin resistance (HOMA-IR) Values were higher, but non-esterified fatty acids (NEFA) were lower on LSI than on CD. For LSI, areas under the curve (AUC) of TG, chylomicron-cholesterol, apoB and the cholesterol/apoB ratio were increased, whereas AUC-NEFA was lowered. LSI did not modify body weight, hematocrit, fasting plasma cholesterol, glucose, adiponectin, leptin, fibrinogen and factor VII (FVII), and AUC of lipoprotein lipase and of lipoprotein remnants. Conclusion: LSI induced alterations in the plasma lipoproteins and in inflammatory markers that are common features of the metabolic syndrome. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Use of the frontal assessment battery in evaluating executive dysfunction in patients with Huntington`s disease

The frontal assessment battery (FAB) is a bedside cognitive scale designed to measure executive functions. Huntington`s disease (HD) is a neurodegenerative disorder characterized by motor, behavioral, and cognitive dysfunction. The aim of this study was to check the validity of the FAB for the evaluation of cognitive impairment in patients with HD. Forty-one patients diagnosed with HD and 53 healthy controls matched by education, sex and age were evaluated with a validated Brazilian version of the UHDRS, the VFT, the SDMT, the SIT, the MMSE, and the FAB. The diagnosis of HD was made by DNA analysis. FAB scores were lower in patients than in the controls (p < 0.001) and had significant correlations with the VFT (r = 0.79; p < 0.05), the SDMT (r = 0.80; p < 0.05), the SIT (r = 0.72; p < 0.05), the MMSE (r = 0.83; p < 0.05), the FCS (r = 0.79; p < 0.05) and the motor section of the UHDRS (r = -0.80; p < 0.05). The FAB differentiated between HD patients in the initial and later stages of the disease. The one-year longitudinal evaluation revealed a global trend toward a worsening in the second score of the FAB. The results demonstrate that the FAB presents good internal consistency and also convergent and discriminative validity; therefore it is a useful scale to assess executive functions and to evaluate cognitive impairment in patients with HD.

Vitrification with Glutamine Improves Maturation Rate of Vitrified/Warmed Immature Bovine Oocytes

Contents The current study examined the protective effects of l-glutamine and cytochalasin B during vitrification of immature bovine oocytes. Oocyte vitrification solution (PBS supplemented with 10% FCS, 25% EG, 25% DMSO and 0.5 m trehalose) was the vitrification control. Treatments were the addition of 7 mu g/ml cytochalasin B, 80 mm glutamine or both cytochalasin and glutaminine for 30 s. After warming, oocytes were matured in vitro for 24 h, fixed and stained with Hoechst (33342) for nuclear maturation evaluation. l-glutamine improved the vitrified/warmed immature bovine oocytes viability (32.8%), increasing the nuclear maturation rates compared to other treatments and the no treatment vitrified control (17.4%). There was, however, no effect of cytochalasin B on in vitro maturation (14.4%).

Notas sobre o mundo social do livro: a construção do editor e da edição

Personagem-filtro, intérprete, mas também interventor, prescrevendo, legitimando e ordenando o universo tipográfico, o editor surge como figura múltipla e socialmente investida de atributos e práticas mediadoras na sua relação com o dado textual. Produtor de valor e materialidade, o editor inscreve o projecto do livro num espaço social colaborativo de trabalho, o campo da edição. Este artigo procura sistematizar teoricamente alguns tópicos relativos à articulação do editor com a construção social do campo editorial e a edificação da cultura impressa. Empreender semelhante exploração é abdicar forçosamente de uma visão linear, unidimensional e historicamente asséptica do mundo social e cultural do livro, cuja morfologia e suportes conhecem crescentemente os desafios da desmaterialização.

Backside-surface imprinting as a new strategy to generate specific plastic antibody materials

A backside protein-surface imprinting process is presented herein as a novel way to generate specific synthetic antibody materials. The template is covalently bonded to a carboxylated-PVC supporting film previously cast on gold, let to interact with charged monomers and surrounded next by another thick polymer. This polymer is then covalently attached to a transducing element and the backside of this structure (supporting film plus template) is removed as a regular “tape”. The new sensing layer is exposed after the full template removal, showing a high density of re-binding positions, as evidenced by SEM. To ensure that the templates have been efficiently removed, this re-binding layer was cleaned further with a proteolytic enzyme and solution washout. The final material was named MAPS, as in the back-side reading of SPAM, because it acts as a back-side imprinting of this recent approach. It was able to generate, for the first time, a specific response to a complex biomolecule from a synthetic material. Non-imprinted materials (NIMs) were also produced as blank and were used as a control of the imprinting process. All chemical modifications were followed by electrochemical techniques. This was done on a supporting film and transducing element of both MAPS and NIM. Only the MAPS-based device responded to oxLDL and the sensing layer was insensitive to other serum proteins, such as myoglobin and haemoglobin. Linear behaviour between log(C, μg mL−1) versus charged tranfer resistance (RCT, Ω) was observed by electrochemical impedance spectroscopy (EIS). Calibrations made in Fetal Calf Serum (FCS) were linear from 2.5 to 12.5 μg mL−1 (RCT = 946.12 × log C + 1590.7) with an R-squared of 0.9966. Overall, these were promising results towards the design of materials acting close to the natural antibodies and applied to practical use of clinical interest.

Estudio de los mecanismos terapéuticos en la inducción de tolerancia immunológica en un modelo animal de esclerosis múltiple

En una serie de estudios previos demostramos que la infusión de células de médula ósea (MO) modificadas genéticamente para la expresión del autoantígeno MOG40-55 en ausencia de mieloablación inducía tolerancia antígenoespecífica en un modelo murino de esclerosis múltiple. También observamos que este efecto terapéutico no requería injerto hematopoyético. Nos propusimos estudiar si el efecto tolerogénico está inducido por una subpoblación de células generadas durante la transducción de la MO y el papel de las células T reguladoras en la inducción de la tolerancia. Las células de MO fueron cultivadas y transducidas usando medio complementado con 20% FCS y medios condicionados como fuente de stem cell factor (SCF) e IL-3 murinos. Las diferentes poblaciones celulares se separaron por citometría de flujo y se analizó la capacidad supresora de las poblaciones candidatas. Por otro lado se analizó la presencia de células T reguladoras en bazo y SNC de los ratones recuperados después de la infusión de células de MO transducidas. A los cinco días de cultivo, la mayoría de células presentaban fenotipo mieloide (Mac-1+Gr-1low/-:31,9+-10,2%; Mac-1+Gr-1high:26,0+-3,3%). Ambos fenotipos se corresponden con dos subpoblaciones de células mieloides supresoras (MDSC, tipo monocí¬tico y granulocítico respectivamente) descritas recientemente. Se estudió la capacidad de ambas poblaciones para suprimir la respuesta proliferativa específica de esplenocitos frente a MOG40-55 in vitro, observando una mayor capacidad de supresión de las MDSC monocíticas, que se correspondí¬a con niveles significativamente superiores de actividad de las enzimas arginasa-1 y sintasa de óxido nítrico (ambos mecanismos supresores característicos de las MDSC). A los 7 días del tratamiento no se observaron diferencias significativas en el porcentaje de células T reguladoras (Treg y Tr1) entre el grupo tratado (liM) y los grupos de control.