Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations

Objective: A new protocol for fixation and slide preservation was evaluated in order to improve the quality of immunocytochemical reactions on cytology slides. Methods: The quality of immunoreactions was evaluated retrospectively on 186 cytology slides (130 direct smears, 56 cytospins) prepared from different cytology samples. Ninety-three of the slides were air dried, stored at -20 °C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 °C for at least 30 minutes, subsequently protected with polyethylene glycol (PEG) and stored at room temperature (Protocol 2). Immunocytochemical staining, with eight primary antibodies, was performed on a Ventana BenchMark Ultra instrument using an UltraView Universal DAB Detection Kit. The following parameters were evaluated for each immunoreaction: morphology preservation, intensity of specific staining, background and counterstain. The slides were blinded and independently scored by four observers with marks from 0 to 20. Results: The quality of immunoreactions was better on methanol-fixed slides protected with PEG than on air-dried slides stored in the freezer: X¯ = 14.44 ± 3.58 versus X¯ = 11.02 ± 3.86, respectively (P < 0.001). Conclusion: Immediate fixation of cytology slides in cold methanol with subsequent application of PEG is an easy and straightforward procedure that improves the quality of immunocytochemical reactions and allows the storage of the slides at room temperature.

Crianças com multideficiência sem linguagem oral expressiva: formas de comunicação mais utilizadas para formular pedidos

O estudo procurou saber quais as formas de comunicação usadas por cinco crianças com multideficiência, com idades compreendidas entre os três e os seis anos, na formulação de pedidos e se os adultos compreendiam essas formas de comunicação. Os dados foram recolhidos através da observação directa dos comportamentos das crianças e de entrevistas efectuadas aos educadores de infância. Os resultados revelaram que as crianças com multideficiência usavam, basicamente, formas de comunicação não simbólica para formular pedidos e raramente os faziam na ausência dos itens que desejavam. Os pedidos formulados relacionaram-se sobretudo com a regulação dos comportamentos e a interacção social. Os resultados indicaram ainda que os adultos tinham dificuldade em responder positivamente às formas de comunicação usadas pelas crianças, levando-as a ter relativamente poucas oportunidades comunicativas e, consequentemente, escassas oportunidades de aprendizagem.

O impacto do Modelo Touchpoints: das representações do educador de infância à construção de uma parentalidade confiante

Na esteira dos estudos sobre a relação creche-família e sobre o modelo Touchpoints, surgiu o interesse de elaborar uma investigação-ação, na qual se pretende implementar práticas reflexivas Touchpoints e avaliar o seu impacto na satisfação dos pais, das educadoras de infância, no desenvolvimento infantil, na qualidade do contexto educativo, nas rotinas da família e no desenvolvimento das crianças. Para o efeito e assumindo um duplo papel enquanto educadoras de infância e investigadoras, foram convidados a participar neste estudo 22 pais de 23 crianças, com idades compreendidas entre os 13 e os 33 meses, que frequentam as salas de creche onde exercemos a nossa atividade profissional, em duas instituições distintas. Esta investigação assentou em quatro fases de trabalho, de forma a cumprir os objetivos propostos: 1ª) a recolha de dados, antes da formação em Touchpoints, recorrendo aos seguintes instrumentos e observações: Escala de Desenvolvimento Infantil Growing Skills II, observação da interação pais-criança com a escala Care-Index, avaliação da satisfação dos pais e das educadoras com a Parent Caregiver Relationship Scale (PCRS), avaliação do contexto educativo com a ITERS-R, e os Mapas de Rotinas Semanais da família; 2ª) formação Touchpoints com uma equipa interdisciplinar; 3ª) Práticas reflexivas Touchpoints – as educadoras como agentes de mudança refletem sobre as suas práticas com guiões prévios e coaching (supervisão com formadores que visitaram a creche, a equipa educativa e a sala da educadora); 4ª) repetição da recolha de dados no final do projeto. O estudo indicou que o desenvolvimento infantil, as relações pais-filhos, as rotinas familiares, a satisfação pais-educadoras e educadoras-pais, assim como a própria prática das educadoras, mudaram após o treino com as práticas reflexivas em Touchpoints. Embora estes resultados não possam ser generalizados e o controlo das variáveis seja afetada por uma metodologia de investigação-ação (portanto, não possamos inferir que todos estes ganhos se devem às praticas reflexivas Touchpoints), os principais resultados obtidos neste estudo permitem-nos repensar formas de relacionamento entre a creche e a família e sobre o impacto que a prática reflexiva dos educadores de infância pode ter na qualidade destas relações. Julgamos que a presente investigação suscita questões sobre a relação educadores-pais e sobre o contributo das práticas reflexivas em Touchpoints nas práticas de creche.

Amplificação em imunohistoquímica: estudo comparativo de sistemas de polímeros

Desde o início da utilização da imunohistoquímica em anatomia patológica, um dos objetivos tem sido detetar as quantidades mais ínfimas de antigénio, tornando-o visível ao microscópio ótico. Vários sistemas de amplificação têm sido aplicados de forma a concretizar este objetivo, tendo surgido um grupo genérico de métodos simples e que apresentam uma amplificação superior: são os denominados métodos do polímero indireto. Tendo em conta a variedade de métodos disponíveis, o autor propõe-se a comparar a qualidade de quatro sistemas de amplificação, que recorrem ao método do polímero indireto com horseradish peroxidase (HRP). Foram utilizadas lâminas de diferentes tecidos, fixados em formol e incluídos em parafina, nos quais se procedeu à identificação de 15 antigénios distintos. Na amplificação recorreu-se a quatro sistemas de polímero indireto (Dako EnVision+ System – K4006; LabVision UltraVision LP Detection System – TL-004-HD; Leica NovoLink – RE7140-k; Vector ImmPRESS Reagent Kit – MP-7402). A observação microscópica e classificação da imunomarcação obtida foram feitas com base num algoritmo que enquadra intensidade, marcação específica, marcação inespecífica e contraste, num score global que pode tomar valores entre 0 e 25. No tratamento dos dados, para além da estatística descritiva, foi utilizado o teste one-way ANOVA com posthoc de tukey (alfa=0.05). O melhor resultado obtido, em termos de par média/desvio-padrão, dos scores globais foi o do NovoLink (22,4/2,37) e o pior foi o do EnVision+ (17,43/3,86). Verificou-se ainda que existe diferença estatística entre os resultados obtidos pelo sistema NovoLink e os sistemas UltraVision (p=.004), ImmPRESS (p=.000) e EnVision+ (p=.000). Concluiu-se que o sistema que permitiu a obtenção de melhores resultados, neste estudo, foi o Leica NovoLink.

Contributo do iPad® para o desenvolvimento de crianças com necessidades educativas especiais

O estudo analisou o papel que o iPad® pode desempenhar no desenvolvimento de crianças com atraso global do desenvolvimento psicomotor (AGDPM), em contexto terapêutico. Neste sentido procurou explorar o potencial do iPad® para promover o desenvolvimento cognitivo e motor de crianças com AGDPM; descrever o modo como estas reagem à utilização do iPad® e conhecer as perspetivas de pais e profissionais face à utilização do iPad®. Participaram no estudo de caso nove crianças com AGDPM, com uma média de idade de 32 meses, nove pais e um profissional de terapia ocupacional. As crianças encontravam-se em período de intervenção em terapia ocupacional, num contexto hospitalar. Efetuaram-se entrevistas semidiretivas aos pais e ao terapeuta ocupacional para conhecer as suas opiniões, procedeu-se a pesquisa documental para caracterizar as crianças e a observações naturalistas para se caracterizar o envolvimento e atividade e participação das crianças com o iPad®. Os resultados evidenciam reações muito positivas por parte das crianças à utilização do iPad®, salientando-se os elevados níveis de envolvimento com o iPad® e de atividade e participação, mormente na dimensão «experiências sensoriais intencionais». Os pais e o profissional de saúde fizeram um balanço muito positivo da utilização do iPad® pelas crianças. Os resultados sugerem que a utilização do iPad® em contexto terapêutico poderá contribuir para o desenvolvimento cognitivo e motor de crianças com diagnóstico de AGDPM.

A divulgação sobre instrumentos derivados : evidência empírica do setor da indústria português

Mestrado em Contabilidade

Insulin resistance does not play a major role in lipid and apolipoprotein profile in obese women

Background: Obesity is associated with increased atherogenesis through alterations in lipids, among other potential factors. Some of those abnormalities might be mediated by insulin resistance (IR). Aims: To compare lipid and apolipoprotein profile between lean and obese women; to evaluate the influence of IR on lipid and apolipoprotein profile, in obese women. Methods: We studied 112 obese and 100 normal-weight premenopausal women without known cardiovascular disease. Both groups were characterized for anthropometrics and a fasting blood sample was collected for assessment of glucose, insulin, triglycerides, cholesterol (total, LDL and HDL), and apolipoproteins A-I, A-II, B, C-II, C-III, and E; IR was assessed by the homeostatic model assessment (HOMA-IR). We compared lipids between obese and lean women; we looked for correlation of those levels with anthropometrics and IR (independently from anthropometrics) in obese women. Results: Obese women were characterized by mean age=34.6±8.3 years, BMI=43.6±7.9 kg/m2, waist circumference (Wc)=117.5±15.1 cm, and HOMA-IR=4.28±3.5. Lean women (age=34.2±8.3 years, BMI=21.4±1.7 kg/m2, Wc=71.7±5.8 cm, and HOMA-IR=1.21±0.76) presented with significantly lower levels of total cholesterol (P=0.001), LDL-cholesterol (P<0.001), and triglycerides (P<0.001); they presented higher levels of HDL-cholesterol (P<0.001), Apo A-I (P<0.001) and Apo A-II (P=0.037). HOMA-IR showed no significant association with apolipoproteins. HOMA-IR was inversely associated with HDL-cholesterol (P=0.048; r=−0.187) but that association disappeared when we adjusted for waist circumference. Only triglycerides were directly associated with HOMA-IR (P<0.001; r=0.343) independently from anthropometrics. Conclusion: We confirm that obese women present worst lipid and apolipoprotein profile. However, with the exception for triglycerides, insulin resistance per se does not play a major role in lipid and apolipoprotein abnormalities observed in obese women.

Genotoxic assessment in different exposure groups working with antineoplastic agents

Antioneoplastic drugs are widely used in treatment of cancer, and several studies suggest acute and long-term effects associated to antineoplastic drug exposures, namely associating workplace exposure with health effects. Cytokinesis blocked micronucleus (CBMN) assay is one promising short-term genotoxicity assays for human risk assessment and their combination is recommended to monitor populations chronically exposed to genotoxic agents. The aim of this investigation is the genotoxicity assessment in different professionals that handle cytostatics drugs. This research is case-control blinded study constituted by 46 non-exposed subjects and 44 workers that handle antineoplastic drugs, such as pharmacists, pharmacy technicians, and nurses. It was found statistically significant increases in the genotoxicity biomarkers in exposed comparising with controls (p<0.05). The findings address the need for regular biomonitoring of personnel occupationally exposed to these drugs, confirming to an enhanced health risk assessment.

Avoiding healthy cells extinction in a cancer model

We consider a dynamical model of cancer growth including three interacting cell populations of tumor cells, healthy host cells and immune effector cells. For certain parameter choice, the dynamical system displays chaotic motion and by decreasing the response of the immune system to the tumor cells, a boundary crisis leading to transient chaotic dynamics is observed. This means that the system behaves chaotically for a finite amount of time until the unavoidable extinction of the healthy and immune cell populations occurs. Our main goal here is to apply a control method to avoid extinction. For that purpose, we apply the partial control method, which aims to control transient chaotic dynamics in the presence of external disturbances. As a result, we have succeeded to avoid the uncontrolled growth of tumor cells and the extinction of healthy tissue. The possibility of using this method compared to the frequently used therapies is discussed. (C) 2014 Elsevier Ltd. All rights reserved.

Consumo de benzodiazepinas por idosos institucionalizados: inapropriação, qualidade de sono e contributo para a segurança na medicação

A racionalização do uso de medicamentos constitui-se como um fator contribuinte para a melhoria da segurança do doente, no que respeita à segurança da medicação e do medicamento, especialmente quando este é administrado a idosos. A avaliação do uso de medicamentos potencialmente inapropriados no idoso constitui-se como uma medida que concorre para evitar, prevenir ou corrigir eventos adversos associados ao seu uso. As benzodiazepinas (BZD) são uma das classes de medicamentos mais prescritas em idosos. No entanto, e apesar da sua utilidade clínica algumas são consideradas inapropriadas nesta faixa etária por potenciarem o efeito sedativo e aumentar a incidência de quedas e fraturas. A longo prazo, na promoção da qualidade do sono, a sua efetividade é discutível já que a toma de uma benzodiazepina para a resolução de um problema como o sono, muitas vezes pontual, passa a ser um problema crónico de exigência de toma contínua, sem que a qualidade deste seja restabelecida, pondo em risco a segurança do doente.

Biomass production of four Cynara cardunculus clones and lignin composition analysis

Four Cynara cardunculus clones, two from Portugal and two from Spain were studied for biomass production and their lignin was characterized. The clones differed in biomass partitioning: Spanish clones produced more capitula (54.5% vs. 43.9%), and Portuguese clones more stalks (37.2% vs. 25.6%). The heating values (HHV0) of the stalks were similar, ranging from 17.1 to 18.4 MJ/kg. Lignin was studied by analytical pyrolysis (Py-GC/MS(FID)), separately in depithed stalks (stalksDP) and pith. StalksDP had in average higher relative proportions of lignin derived compounds than pith (23.9% vs. 21.8%) with slightly different lignin monomeric composition: pith samples were richer in syringyl units as compared to stalksDP (64% vs. 53%), with S/G ratios of 2.1 and 1.3, respectively. The H:G:S composition was 7:40:53 in stalksDP and 7:29:64 in pith. The lignin content ranged from 18.8% to 25.5%, enabling a differentiation between clones and provenances. © 2015 Elsevier Ltd. All rights reserved.

Liquid-vapor interfaces of patchy colloids

We investigate the liquid-vapor interface of a model of patchy colloids. This model consists of hard spheres decorated with short-ranged attractive sites ("patches") of different types on their surfaces. We focus on a one-component fluid with two patches of type A and nine patches of type B (2A9B colloids), which has been found to exhibit reentrant liquid-vapor coexistence curves and very low-density liquid phases. We have used the density-functional theory form of Wertheim's first-order perturbation theory of association, as implemented by Yu and Wu [J. Chem. Phys. 116, 7094 (2002)], to calculate the surface tension, and the density and degree of association profiles, at the liquid-vapor interface of our model. In reentrant systems, where AB bonds dominate, an unusual thickening of the interface is observed at low temperatures. Furthermore, the surface tension versus temperature curve reaches a maximum, in agreement with Bernardino and Telo da Gama's mesoscopic Landau-Safran theory [Phys. Rev. Lett. 109, 116103 (2012)]. If BB attractions are also present, competition between AB and BB bonds gradually restores the monotonic temperature dependence of the surface tension. Lastly, the interface is "hairy," i.e., it contains a region where the average chain length is close to that in the bulk liquid, but where the density is that of the vapor. Sufficiently strong BB attractions remove these features, and the system reverts to the behavior seen in atomic fluids.

Vertex component analysis: a geometric-based approach to unmix hyperspectral data

Hyperspectral remote sensing exploits the electromagnetic scattering patterns of the different materials at specific wavelengths [2, 3]. Hyperspectral sensors have been developed to sample the scattered portion of the electromagnetic spectrum extending from the visible region through the near-infrared and mid-infrared, in hundreds of narrow contiguous bands [4, 5]. The number and variety of potential civilian and military applications of hyperspectral remote sensing is enormous [6, 7]. Very often, the resolution cell corresponding to a single pixel in an image contains several substances (endmembers) [4]. In this situation, the scattered energy is a mixing of the endmember spectra. A challenging task underlying many hyperspectral imagery applications is then decomposing a mixed pixel into a collection of reflectance spectra, called endmember signatures, and the corresponding abundance fractions [8–10]. Depending on the mixing scales at each pixel, the observed mixture is either linear or nonlinear [11, 12]. Linear mixing model holds approximately when the mixing scale is macroscopic [13] and there is negligible interaction among distinct endmembers [3, 14]. If, however, the mixing scale is microscopic (or intimate mixtures) [15, 16] and the incident solar radiation is scattered by the scene through multiple bounces involving several endmembers [17], the linear model is no longer accurate. Linear spectral unmixing has been intensively researched in the last years [9, 10, 12, 18–21]. It considers that a mixed pixel is a linear combination of endmember signatures weighted by the correspondent abundance fractions. Under this model, and assuming that the number of substances and their reflectance spectra are known, hyperspectral unmixing is a linear problem for which many solutions have been proposed (e.g., maximum likelihood estimation [8], spectral signature matching [22], spectral angle mapper [23], subspace projection methods [24,25], and constrained least squares [26]). In most cases, the number of substances and their reflectances are not known and, then, hyperspectral unmixing falls into the class of blind source separation problems [27]. Independent component analysis (ICA) has recently been proposed as a tool to blindly unmix hyperspectral data [28–31]. ICA is based on the assumption of mutually independent sources (abundance fractions), which is not the case of hyperspectral data, since the sum of abundance fractions is constant, implying statistical dependence among them. This dependence compromises ICA applicability to hyperspectral images as shown in Refs. [21, 32]. In fact, ICA finds the endmember signatures by multiplying the spectral vectors with an unmixing matrix, which minimizes the mutual information among sources. If sources are independent, ICA provides the correct unmixing, since the minimum of the mutual information is obtained only when sources are independent. This is no longer true for dependent abundance fractions. Nevertheless, some endmembers may be approximately unmixed. These aspects are addressed in Ref. [33]. Under the linear mixing model, the observations from a scene are in a simplex whose vertices correspond to the endmembers. Several approaches [34–36] have exploited this geometric feature of hyperspectral mixtures [35]. Minimum volume transform (MVT) algorithm [36] determines the simplex of minimum volume containing the data. The method presented in Ref. [37] is also of MVT type but, by introducing the notion of bundles, it takes into account the endmember variability usually present in hyperspectral mixtures. The MVT type approaches are complex from the computational point of view. Usually, these algorithms find in the first place the convex hull defined by the observed data and then fit a minimum volume simplex to it. For example, the gift wrapping algorithm [38] computes the convex hull of n data points in a d-dimensional space with a computational complexity of O(nbd=2cþ1), where bxc is the highest integer lower or equal than x and n is the number of samples. The complexity of the method presented in Ref. [37] is even higher, since the temperature of the simulated annealing algorithm used shall follow a log( ) law [39] to assure convergence (in probability) to the desired solution. Aiming at a lower computational complexity, some algorithms such as the pixel purity index (PPI) [35] and the N-FINDR [40] still find the minimum volume simplex containing the data cloud, but they assume the presence of at least one pure pixel of each endmember in the data. This is a strong requisite that may not hold in some data sets. In any case, these algorithms find the set of most pure pixels in the data. PPI algorithm uses the minimum noise fraction (MNF) [41] as a preprocessing step to reduce dimensionality and to improve the signal-to-noise ratio (SNR). The algorithm then projects every spectral vector onto skewers (large number of random vectors) [35, 42,43]. The points corresponding to extremes, for each skewer direction, are stored. A cumulative account records the number of times each pixel (i.e., a given spectral vector) is found to be an extreme. The pixels with the highest scores are the purest ones. N-FINDR algorithm [40] is based on the fact that in p spectral dimensions, the p-volume defined by a simplex formed by the purest pixels is larger than any other volume defined by any other combination of pixels. This algorithm finds the set of pixels defining the largest volume by inflating a simplex inside the data. ORA SIS [44, 45] is a hyperspectral framework developed by the U.S. Naval Research Laboratory consisting of several algorithms organized in six modules: exemplar selector, adaptative learner, demixer, knowledge base or spectral library, and spatial postrocessor. The first step consists in flat-fielding the spectra. Next, the exemplar selection module is used to select spectral vectors that best represent the smaller convex cone containing the data. The other pixels are rejected when the spectral angle distance (SAD) is less than a given thresh old. The procedure finds the basis for a subspace of a lower dimension using a modified Gram–Schmidt orthogonalizati on. The selected vectors are then projected onto this subspace and a simplex is found by an MV T pro cess. ORA SIS is oriented to real-time target detection from uncrewed air vehicles using hyperspectral data [46]. In this chapter we develop a new algorithm to unmix linear mixtures of endmember spectra. First, the algorithm determines the number of endmembers and the signal subspace using a newly developed concept [47, 48]. Second, the algorithm extracts the most pure pixels present in the data. Unlike other methods, this algorithm is completely automatic and unsupervised. To estimate the number of endmembers and the signal subspace in hyperspectral linear mixtures, the proposed scheme begins by estimating sign al and noise correlation matrices. The latter is based on multiple regression theory. The signal subspace is then identified by selectin g the set of signal eigenvalue s that best represents the data, in the least-square sense [48,49 ], we note, however, that VCA works with projected and with unprojected data. The extraction of the end members exploits two facts: (1) the endmembers are the vertices of a simplex and (2) the affine transformation of a simplex is also a simplex. As PPI and N-FIND R algorithms, VCA also assumes the presence of pure pixels in the data. The algorithm iteratively projects data on to a direction orthogonal to the subspace spanned by the endmembers already determined. The new end member signature corresponds to the extreme of the projection. The algorithm iterates until all end members are exhausted. VCA performs much better than PPI and better than or comparable to N-FI NDR; yet it has a computational complexity between on e and two orders of magnitude lower than N-FINDR. The chapter is structure d as follows. Section 19.2 describes the fundamentals of the proposed method. Section 19.3 and Section 19.4 evaluate the proposed algorithm using simulated and real data, respectively. Section 19.5 presents some concluding remarks.

Ventilação térmica "Féria" - tipo Shunt

As instalações de ventilação em estudo destinam-se a realizar o arejamento geral e permanente de habitações - pelo menos durante o período do ano em que a temperatura exterior obriga a manter as janelas fechadas - e a assegurar a evacuação de gases e fumos resultantes da combustão em aparelhos de aquecimento por queima (esquentadores e fogões (de uso corrente nas habitações)).