External quality assessment in the voluntary counseling and testing centers in the Brazilian Amazon using dried tube specimens: results of an effectiveness evaluation

INTRODUCTION : In 2011, the Brazilian Ministry of Health rolled out a program for the external quality assessment of rapid human immunodeficiency virus (HIV) tests using the dried tube specimen (DTS) method (EQA-RT/DTS-HIV). Our objective was to evaluate the implementation of this program at 71 voluntary counseling and testing centers (VCTCs) in the Brazilian Legal Amazonian area one year after its introduction. METHODS : Quantitative and qualitative study that analyzed secondary data and interviews with healthcare workers (HCWs) (n=39) and VCTC coordinators (n=32) were performed. The assessment used 18 key indicators to evaluate the three dimensions of the program's logical framework: structure, process, and result. Each indicator was scored from 1-4, and the aggregate results corresponding to the dimensions were expressed as proportions. The results were compared to the perceptions of the HCWs and coordinators regarding the EQA-RT/DTS-HIV program. RESULTS: The aggregate scores for the three dimensions of structure, process, and result were 91.7%, 78.6%, and 95%, respectively. The lowest score in each dimension corresponded to a different indicator: access to Quali-TR online system 39% (structure), registration in Quali-TR online system 38.7% (process), and VCTC completed the full process in the program's first round 63.4% (result). Approximately 36% of the HCWs and 52% of the coordinators reported enhanced trust in the program for its rapid HIV testing performance. CONCLUSIONS: All three program dimensions exhibited satisfactory results (>75%). Nevertheless, the study findings highlight the need to improve certain program components. Additionally, long-term follow-ups is needed to provide a more thorough picture of the process for external quality assessment.

Implementação da Equipa de Emergência Médica Intra-hospitalar nos hospitais do Serviço Nacional de Saúde : análise de custo-efetividade perante a incidência da paragem cárdio-respiratória intra-hospitalar

RESUMO - Introdução: No âmbito das emergências intra-hospitalares investigou-se a hipótese da presença da Equipa Emergência Médica Intra-hospitalar (EEMI) (DGS, 2010) num Centro Hospitalar (CH), contribuir para a redução do número de mortos por Paragem Cárdiorespiratória (PCR) intra-hospitalar, quando comparado com outro CH dotado de uma equipa tradicional de resposta à PCR. Metodologia: Tratou-se de um estudo observacional, retrospetivo (2010 a 2014), com base nos dados do Grupo de Diagnóstico Homogéneo (GDH), analisado numa perspetiva de custo-efetividade no impacto sobre incidência de PCR e taxa de mortalidade. Resultados: Observou-se que o CH com EEMI apresentou uma Redução Risco Absoluto (RRA) de 9,01% de morte por PCR. A taxa de mortalidade calculada foi de 2,82 casos por 1000 episódios de internamento em que a incidência de PCR foi de 28,24 casos por cada 10 000 habitantes, duas vezes menor que CH em comparação. Quando introduzidas manobras de Ressuscitação Cárdiopulmonar (RCP), o mesmo CH teve um maior número de PCR revertidas, com uma taxa de mortalidade 2 vezes menor que o CH sem EEMI. Conclusão: Resultados demonstraram que os dois CH apresentaram riscos diferentes, em que a probabilidade do doente hospitalizado de morrer após ocorrência de PCR foi menor no grupo exposto à EEMI, com OR = 0,496 [IC 95% (0,372 a 0,662)] para dados populacionais (p = 0,0013), e OR = 0,618 [IC 95% (0,298 a 1,281)] para dados individuais, (p = 0,194). Face a melhores resultados em Saúde, considerou-se a implementação da EEMI, uma medida custo-efetiva, uma vez que o principal requisito traduz-se por reorganização das equipas tradicionais para uma vertente de prevenção da PCR.

Cellulose-based composites as functional conductive materials for printed electronics

In this work, cellulose-based electro and ionic conductive composites were developed for application in cellulose based printed electronics. Electroconductive inks were successfully formulated for screen-printing using carbon fibers (CFs) and multi-walled carbon nanotubes (MWCNTs) as conductive functional material and cellulose derivatives working as binder. The formulated inks were used to fabricate conductive flexible and disposable electrodes on paper-based substrates. Interesting results were obtained after 10 printing passes and drying at RT of the ink with 10 % wt. of pristine CFs and 3% wt. of carboxymethyl cellulose (CMC), exhibiting a resistivity of 1.03 Ωcm and a resolution of 400 μm. Also, a resistivity of 0.57 Ωcm was obtained for only one printing pass using an ink based on 0.5 % wt. MWCNTs and 3 % wt. CMC. It was also demonstrated that ionic conductive cellulose matrix hydrogel can be used in electrolyte-gated transistors (EGTs). The electrolytes revealed a double layer capacitance of 12.10 μFcm-2 and ionic conductivity of 3.56x10-7 Scm-1. EGTs with a planar configuration, using sputtered GIZO as semiconducting layer, reached an ON/OFF ratio of 3.47x105, a VON of 0.2 V and a charge carrier mobility of 2.32 cm2V-1s-1.

Avaliação do envolvimento das bombas de efluxo na fármaco-resistência ao Praziquantel em Schistosoma Mansonii

O Praziquantel (PZQ) é o fármaco de primeira linha no tratamento da schistosomose, com alta taxa de cura e sem efeitos secundários significativos. Têm sido reportados cada vez mais casos de resistência ou de aumento de tolerância a este fármaco, aumentando as preocupações de emergência de estirpes resistentes ao PZQ. O Verapamil, um bloqueador de canais de cálcio, inibe o fluxo de fármaco activo e tem sido descrito como um bom inibidor das glicoproteínas-P (P-gp), sendo, por isso, utilizado em diversos estudos de fármaco-resistência. No laboratório de Helmintologia da Unidade de Ensino e Investigação em Parasitologia Médica do Instituto de Higiene e Medicina Tropical, foi seleccionada uma linha de S. mansoni, resistente a 800 mg/Kg de PZQ, após pressão de fármaco constante e crescente ao longo de vários ciclos. Para confirmar a existência de diferenças polimórficas entre a estirpes sensível e resistente de S. mansoni, extraiu-se o DNA de parasitas adultos de ambas as estirpes de S. mansoni e analisou-se por Random Amplified Polymorphic DNAPolymerase Chain Reaction (RAPD-PCR). As diferenças polimórficas entre a estirpe sensível e a resistente foram observadas e calculou-se o coeficiente de similaridade (Dice’s coefficient). Após confirmar a existência de polimorfismos entre as duas estirpes, a atividade das bombas de efluxo foi avaliada em ambas as estirpes. A avaliação foi realizada num ensaio de acumulação usando o composto Brometo de Etídio na presença e ausência de Verapamil. O papel das bombas de efluxo na resistência ao PZQ, foi ainda investigado comparando a resposta dos parasitas da estirpe sensível e resistente ao fármaco na ausência e na presença de diferentes doses de Verapamil, em cultura in vitro. Os resultados obtidos foram reforçados comparando os níveis e expressão do gene SmMDR2 em ambas as estirpes isogénicas por Real-Time PCR (qPCR). A estirpe resistente de S. mansoni, necessitou de concentrações mais elevadas de inibidor quando comparada com a estirpe sensível para obter níveis significativos de fluorescência de Brometo de Etídio. A cultura in vitro mostrou uma dose letal de PZQ mais elevada na estirpe resistente do que na estirpe sensível na ausência de Verapamil. Na presença de Verapamil houve uma redução na dose letal de PZQ nos machos de ambas as estirpes, sendo esta redução mais acentuada nos machos da estirpe resistente. As fêmeas não mostraram alterações significativas na dose letal de PZQ na presença e ausência e inibidor. Os resultados foram reforçados pela observação dos níveis do gene SmMDR2, onde os machos da estirpe resistente mostraram ter os maiores níveis de expressão e pelo aumento de expressão nos machos de ambas as estirpes após exposição ao PZQ. As fêmeas de ambas as estirpes não tiveram diferenças significativas na expressão do gene após exposição ao PZQ e as fêmeas da estirpe resistente tiveram mostraram ter os níveis de expressão do gene SmMDR2 mais baixos entre os machos e fêmeas de ambas as estirpes. Os resultados obtidos neste trabalho mostraram que os machos da estirpe resistente têm maior atividade de bombas de efluxo do que os machos da estirpe sensível e que as bombas P-gp estão envolvidas na resposta e no aumento de tolerância dos machos de S. mansoni ao PZQ.

Modulation of the secretome of hBMSCs by tailoring the macromolecular gradient in hydrogels to generate tissue-to-tissue interfaces

Tissue-to-tissue interfaces are commonly present in all tissues exhibiting structural, biological and chemical gradients serving a wide range of physiological functions. These interfaces are responsible for mediation of load transfer between two adjacent tissues. They are also important structures in sustaining the cellular communications to retain tissueâ s functional integration and homeostasis. [1] All cells have the capacity to sense and respond to physical and chemical stimulus and when cultured in three-dimensional (3D) environments they tend to perform their function better than in two-dimensional (2D) environments. Spatial and temporal 3D gradient hydrogels better resemble the natural environment of cells in mimicking their extracellular matrix. [2] In this study we hypothesize that differential functional properties can be engineered by modulation of macromolecule gradients in a cell seeded threedimensional hydrogel system. Specifically, differential paracrine secretory profiles can be engineered using human Bone Marrow Stem Cells (hBMSCâ s). Hence, the specific objectives of this study are to: assemble the macromolecular gradient hydrogels to evaluate the suitablity for hBMSCâ s encapsulation by cellular viability and biofunctionality by assessing the paracrine secretion of hBMSCâ s over time. The gradient hydrogels solutions were prepared by blend of macromolecules in one solution such as hyaluronic (HA) acid and collagen (Col) at different ratios. The gradient hydrogels were fabricated into cylindrical silicon moulds with higher ratio solutions assembled at the bottom of the mould and adding the two solutions consecutively on top of each other. The labelling of the macromolecules was performed to confirm the gradient through fluorescence microscopy. Additionally, AFM was conducted to assess the gradient hydrogels stiffness. Gradient hydrogels characterization was performed by HA and Col degradation assay, degree of crosslinking and stability. hBMSCâ s at P3 were encapsulated into each batch solution at 106 cells/ml solution and gradient hydrogels were produced as previously described. The hBMSCâ s were observed under confocal microscopy to assess viability by Live/Dead® staining. Cellular behaviour concerning proliferation and matrix deposition was also performed. Secretory cytokine measurement for pro-inflammatory and angiogenesis factors was carried out using ELISA. At genomic level, qPCR was carried out. The 3D gradient hydrogels platform made of different macromolecules showed to be a suitable environment for hBMSCâ s. The hBMSCâ s gradient hydrogels supported high cell survival and exhibited biofunctionality. Besides, the 3D gradient hydrogels demonstrated differentially secretion of pro-inflammatory and angiogenic factors by the encapsulated hBMSCâ s. References: 1. Mikos, AG. et al., Engineering complex tissues. Tissue Engineering 12,3307, 2006 2. Phillips, JE. et al., Proc Natl Acad Sci USA, 26:12170-5, 2008

Establishing guidelines for obtaining optimal cell sources to use in tendon tissue engineering strategies

Cell-based approaches in tissue engineering (TE) have been barely explored for the treatment of tendon and ligament (T/L) tissues, requiring the establishment of a widely available cell source with tenogenic potential. As T/L cells are scarce, stem cells may provide a good alternative. Understanding how resident cells behave in vitro, might be useful for recapitulating the tenogenic potential of stem cells for tendon TE applications. Therefore, we propose to isolate and characterize human T/L-derived cells (hTDCs and hLDCs) and compare their regenerative potential with stem cells from adipose tissue (hASCs) and amniotic fluid (hAFSCs)(1). T/L cells were isolated using different procedures and stem cells isolated as described elsewhere(1). Moreover, T/L cells were stimu- lated into the three mesenchymal lineages, using standard differentia- tion media. Cells were characterized for the typical stem cell markers as well as T/L related markers, namely tenascin-C, collagen I and III, decorin and scleraxis, using different complementary techniques such as real time RT-PCR, immunocytochemistry and flow cytometry. No differences were observed between T/L in gene expression and protein deposition. T/L cells were mostly positive for stem ness markers (CD73/CD90/CD105), and have the potential to differentiate towards osteogenesis, chondrogenesis and adipogenesis, demonstrated by the positive staining for AlizarinRed, SafraninO, ToluidineBlue and OilRed. hASCs and hAFSCs exhibit positive expression of all tenogenic mark- ers, although at lower levels than hTDCs and hLDCs. Nevertheless, stem cells availability is key factor in TE strategies, despite that it’s still required optimization to direct their tenogenic phenotype.

Cryopreservation of Cell Sheets of Adipose Stem Cells: Limitations and Successes

Cell Sheets of hASCs (hASCs-CS) have been previously proposed for wound healing applications(1, 2) and despite the concern for production time reduction, the possibility of having these hASCs-CS off-the-shelf is appealing. The goal of this work was to define a cryopreservation methodology allowing to preserve cells viability and the properties CS matrix. hASCs-CS obtained from three different donors were created in UP-cell thermoresponsive dishes(Nunc, Germany) as previously reported(1,2). Different cryopreservation conditions were considered: i)FBS plus DMSO(5% and10%); ii)0.4M of Trehalose plus DMSO (5% and 10%); iii)cryosolution PLL (Akron Biotech, USA); and iv)vitrification. The cryopreservation effect was first assessed for cellular viability by flow cytometry using 7-AAD, and after dissociating the hASCs-CS with collagenase and trypsin-EDTA 0.25%. The expression (RT-PCR) and deposition (western blot and immunocytochemistry) of collagen type I, laminin and fibronectin, and the organization (TEM) of the extracellular matrix was further assessed before and after hASCs-CS cryopreservation to determine a potential effect of the method over matrix composition and integrity. The obtained results confirmed that cell viability is affected by the cryopreservation methodology, as shown before for different CS(3). Interestingly, the matrix properties were not significantly altered and the typical cell sheetâ s easiness of manipulation for transplantation was not lost.

Relatório de atividade profissional

Dissertação de mestrado em Ciências - Formação Contínua de Professores (área de especialização em Biologia e Geologia)

Relatório de atividade profissional

Dissertação de mestrado em Ciências – Formação Contínua de Professores (área de especialização em Biologia e Geologia)

The influence of kaolin application on key metabolic pathways associated with grape berry quality: a molecular and biochemical analysis

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Hot-dip galvanized steel dip-coatedwith ureasilicate hybrid in simulated concrete pore solution: Assessment of coating morphology and corrosion protection efficiency

The barrier effect and the performance of an organic–inorganic hybrid (OIH) sol–gel coating are highlydependent on the coating deposition method as well as processing conditions. In this work, studies onthe influence of experimental parameters using the dip coating method were performed. Factors suchas residence time (Rt), a curing step between each dip step and the number of layers of sol–gel OIHfilms deposited on HDGS to prevent corrosion in highly alkaline environments were studied. These OIHcoatings were obtained using a functionalized siloxane, 3-isociantepropyltriethoxysilane that reactedwith a diamino-functionalized oligopolymer (Jeffamine®D-230). The barrier efficiency of OIH coatings insimulated concrete pore solutions (SCPS) was assessed in the first moments of contact, by electrochemicalimpedance spectroscopy and potentiodynamic methods. The durability and stability of the OIH coatings inSCPS was monitored during eight days by macrocell current density. The morphological characterizationof the surface was performed by scanning electronic microscopy before and after exposure to SCPS.Glow discharge optical emission spectroscopy was used to obtain quantitative composition profiles toinvestigate the thickness of the OIH coatings as a function of the number of layers deposited and theinfluence of the Rt in the coating thickness.

Relatório de atividade profissional

Dissertação de mestrado em Ciências – Formação Contínua de Professores (área de especialização em Biologia e Geologia)

Adipogenic cell sheets to recreate in vitro adipose tissue microenvironments

Cell sheet (CS) engineering, taking advantage of cellular self-matrix organized as in native tissue, has been largely explored, including by us, for different purposes [1â 3]. Herein we propose for the ï¬ rst time, the use of human adipose stem cells (hASCs)-derived CS to create adipose tissue analogues with different levels of maturation. hASCs were cultured on UpCellTM thermo-responsive dishes for 1, 3 and 5 days under basal conditions previously established by us [3]. The inï¬ uence of pre-differentiation time and respective cell number, over CS stability and differentiation was assessed. Mechanically robust CS were only obtained with 5 days pre-differentiation period. Adipogenesis was followed along the culture assessing the variation of expression of mesenchymal (CD73, CD105 but not CD90) and adipogenic (PPARg, FABP4 and LPL) markers by ï¬ ow cytometry, immunocytochemistry and RT-PCR. Increased ratio of differentiated cells was achieved for longer pre-differentiation periods, while maturation degree was modulated by the maintenance medium. Independently of the overall CS differentiation/maturation level, 3D constructs were fabricated by stacking and further culturing 3 CS. Thus, by varying the culture conditions, different 3D adipose tissue-like microenvironments were recreated, enabling future development of new tissue engineering strategies, as well as further study of adipose tissue role in the regeneration of different tissues.

MMP-9/RECK imbalance: a mechanism associated with high-grade cervical lesions and genital infection by Human Papillomavirus (HPV) and Chlamydia trachomatis

"Manuscript"

Reforma psiquiátrica e serviços residenciais terapêuticos

OBJETIVO: Apresentar os principais aspectos da reforma psiquiátrica ocorrida em diversos países e no Brasil, destacar a política de reinserção social dos pacientes e descrever os procedimentos da desospitalização ocorridos em Barbacena, Minas Gerais, com ênfase nas residências terapêuticas e nas intervenções de reabilitação psicossocial. MÉTODO: Pesquisa bibliográfica realizada nas fontes eletrônicas Medline e Lilacs (1990 a 2006), com os seguintes descritores: deinstitutionalization, psychiatric reform, community-based treatment, psychosocial rehabilitation. Descrição do processo de implantação das residências terapêuticas (RT) por meio de consulta aos documentos da Prefeitura de Barbacena e de observação direta das residências. RESULTADOS: A maioria dos trabalhos selecionados indicou que pacientes desospitalizados apresentam melhora na autonomia, na interação social, no nível global de funcionamento e na qualidade de vida. A desospitalização implica a existência de serviços comunitários com diversos níveis de atenção e requer a presença de profissionais qualificados e programas de reabilitação efetivos. Em Barbacena, o processo de desospitalização teve início em 2000. Atualmente existem 24 RTs no município. CONCLUSÃO: Apesar das dificuldades na sua implantação, os tratamentos de base comunitária tornaram-se o modelo dominante de cuidados psiquiátricos. Para os pacientes desospitalizados, as RTs têm importante papel no processo de reinserção social.