Texturas e propriedades reológicas dos minérios de ferro do quadrilátero ferrífero (MG) e sua utilização em beneficiamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Contração de polimerização e expansão higroscópica de resinas compostas: análise pela técnica de video-imagem

To evaluate the volumetric changes due to polymerization and thermocycling on different resin-composites. Methods: Thirteen A2 Universal Dentin shade resin-composites (n = 10) from eight manufacturers were evaluated (4Seasons, Grandio, Venus, Amelogen Plus, P90, Z350, Esthet-X, Amaris, Vita-l-escence, Natural-Look, Charisma, Z250 and Opallis). The polymerization shrinkage percentage (PS) was calculated using an image measurement device (ACUVOL - Bisco Dental). Equal volumes of material, standardized by a semisphere polyurethane matrix (d = 3mm) were used and, after 5 minutes of relaxation, the baseline volume measurements were obtained with 18 J of energy dose from the LED light-curing unit. Measurements were obtained after 5 minutes and PS values calculated. Specimens were stored in a drydark environment for 24 hours and re-measured. Specimens were then thermocycled in distilled water between 5oC and 55oC for 20,000 cycles, subjected to another volume measurement at 5,000 cycle intervals. Specimens were gently dried prior to each measurement. Results: Repeated measurements were made using ANOVA (α = 0.05) showed that all resin-composite volumes were influenced by the number of cycles. Volumes at 5 minutes post-polymerization (12.47 ± 0.08) were significantly lower than those at baseline (12.80 ± 0.09). Volumes at 24 hours (12.43 ± 0.19) were insignificantly lower than those at 5 minutes postpolymerization. With regards to the impact of thermocycling, all specimens showed statistically significant increases in volume after 5,000 cycles (13.04 ± 0.22). Although statistically different from those after 5,000 cycles, there was no statistically significant difference between volumes measured at 10,000 (12.87±0.21), 15,000 (12.92±0.24), and 20,000 (12.84±0.23) cycles. Conclusion: According to the video-imaging analysis, thermocycling caused a significant expansion in resin-composites tested, the volume increase was not able to...

Análise quantitativa das descargas epileptiformes generalizadas e da neuroimagem de pacientes com epilepsia generalizada idiopática

Pós-graduação em Fisiopatologia em Clínica Médica - FMB

Regulation of SRC-3 localization and dynamics by phosphorylation during ERα-dependent transcriptional activation

Transcription is controlled by promoter-selective transcriptional factors (TFs), which bind to cis-regulatory enhancers elements, termed hormone response elements (HREs), in a specific subset of genes. Regulation by these factors involves either the recruitment of coactivators or corepressors and direct interaction with the basal transcriptional machinery (1). Hormone-activated nuclear receptors (NRs) are well characterized transcriptional factors (2) that bind to the promoters of their target genes and recruit primary and secondary coactivator proteins which possess many enzymatic activities required for gene expression (1,3,4). In the present study, using single-cell high-resolution fluorescent microscopy and high throughput microscopy (HTM) coupled to computational imaging analysis, we investigated transcriptional regulation controlled by the estrogen receptor alpha (ERalpha), in terms of large scale chromatin remodeling and interaction with the associated coactivator SRC-3 (Steroid Receptor Coactivator-3), a member of p160 family (28) primary coactivators. ERalpha is a steroid-dependent transcriptional factor (16) that belongs to the NRs superfamily (2,3) and, in response to the hormone 17-ß estradiol (E2), regulates transcription of distinct target genes involved in development, puberty, and homeostasis (8,16). ERalpha spends most of its lifetime in the nucleus and undergoes a rapid (within minutes) intranuclear redistribution following the addition of either agonist or antagonist (17,18,19). We designed a HeLa cell line (PRL-HeLa), engineered with a chromosomeintegrated reporter gene array (PRL-array) containing multicopy hormone response-binding elements for ERalpha that are derived from the physiological enhancer/promoter region of the prolactin gene. Following GFP-ER transfection of PRL-HeLa cells, we were able to observe in situ ligand dependent (i) recruitment to the array of the receptor and associated coregulators, (ii) chromatin remodeling, and (iii) direct transcriptional readout of the reporter gene. Addition of E2 causes a visible opening (decondensation) of the PRL-array, colocalization of RNA Polymerase II, and transcriptional readout of the reporter gene, detected by mRNA FISH. On the contrary, when cells were treated with an ERalpha antagonist (Tamoxifen or ICI), a dramatic condensation of the PRL-array was observed, displacement of RNA Polymerase II, and complete decreasing in the transcriptional FISH signal. All p160 family coactivators (28) colocalize with ERalpha at the PRL-array. Steroid Receptor Coactivator-3 (SRC-3/AIB1/ACTR/pCIP/RAC3/TRAM1) is a p160 family member and a known oncogenic protein (4,34). SRC-3 is regulated by a variety of posttranslational modifications, including methylation, phosphorylation, acetylation, ubiquitination and sumoylation (4,35). These events have been shown to be important for its interaction with other coactivator proteins and NRs and for its oncogenic potential (37,39). A number of extracellular signaling molecules, like steroid hormones, growth factors and cytokines, induce SRC-3 phosphorylation (40). These actions are mediated by a wide range of kinases, including extracellular-regulated kinase 1 and 2 (ERK1-2), c-Jun N-terminal kinase, p38 MAPK, and IkB kinases (IKKs) (41,42,43). Here, we report SRC-3 to be a nucleocytoplasmic shuttling protein, whose cellular localization is regulated by phosphorylation and interaction with ERalpha. Using a combination of high throughput and fluorescence microscopy, we show that both chemical inhibition (with U0126) and siRNA downregulation of the MAP/ERK1/2 kinase (MEK1/2) pathway induce a cytoplasmic shift in SRC-3 localization, whereas stimulation by EGF signaling enhances its nuclear localization by inducing phosphorylation at T24, S857, and S860, known partecipants in the regulation of SRC-3 activity (39). Accordingly, the cytoplasmic localization of a non-phosphorylatable SRC-3 mutant further supports these results. In the presence of ERalpha, U0126 also dramatically reduces: hormone-dependent colocalization of ERalpha and SRC-3 in the nucleus; formation of ER-SRC-3 coimmunoprecipitation complex in cell lysates; localization of SRC-3 at the ER-targeted prolactin promoter array (PRL-array) and transcriptional activity. Finally, we show that SRC-3 can also function as a cotransporter, facilitating the nuclear-cytoplasmic shuttling of estrogen receptor. While a wealth of studies have revealed the molecular functions of NRs and coregulators, there is a paucity of data on how these functions are spatiotemporally organized in the cellular context. Technically and conceptually, our findings have a new impact upon evaluating gene transcriptional control and mechanisms of action of gene regulators.

Surgical hip dislocation for osteochondral transplantation as a salvage procedure for a femoral head impaction fracture

Obturator anterior hip dislocation is very rare. Poor results are described in patients with additional large transchondral fractures and treatment of these injuries remains challenging. Appropriate treatment recommendations are missing in the literature. This case report introduces surgical hip dislocation for osteochondral autograft transplantation with graft harvest from the nonweightbearing area of the head-neck junction as a salvage procedure in a large femoral head defect. We report the treatment and outcome of a 48-year-old man who sustained an anterior dislocation of the left hip after a motorcycle accident. After initial closed reduction in the emergency room, imaging analysis revealed a large osteochondral defect of the femoral head within the weightbearing area (10 × 20 mm, depth: 5 mm). The hip was exposed with a surgical hip dislocation using a trochanteric osteotomy. An osteochondral autograft was harvested from a nonweightbearing area of the femoral head and transferred into the defect. The patient was prospectively examined clinically and radiologically. Two years postoperatively, the patient was free of pain and complaints. The function of the injured hip was comparable to that of the contralateral, healthy hip and showed satisfying radiologic results. Surgical hip dislocation with a trochanteric flip osteotomy is a simple, one-step technique that allows full inspection of the hip to treat osteochondral femoral defects by osteochondral transplantation. The presented technique, used as a salvage procedure in a large femoral head defect, yielded good clinical and satisfying radiologic outcomes at the midterm.

Development of a novel fMRI compatible visual perception prototype battery to test older people with and without dementia

Background: Visuoperceptual deficits in dementia are common and can reduce quality of life. Testing of visuoperceptual function is often confounded by impairments in other cognitive domains and motor dysfunction. We aimed to develop, pilot, and test a novel visuocognitive prototype test battery which addressed these issues, suitable for both clinical and functional imaging use. Methods: We recruited 23 participants (14 with dementia, 6 of whom had extrapyramidal motor features, and 9 age-matched controls). The novel Newcastle visual perception prototype battery (NEVIP-B-Prototype) included angle, color, face, motion and form perception tasks, and an adapted response system. It allows for individualized task difficulties. Participants were tested outside and inside the 3T functional magnetic resonance imaging (fMRI) scanner. Functional magnetic resonance imaging data were analyzed using SPM8. Results: All participants successfully completed the task inside and outside the scanner. Functional magnetic resonance imaging analysis showed activation regions corresponding well to the regional specializations of the visual association cortex. In both groups, there was significant activity in the ventral occipital-temporal region in the face and color tasks, whereas the motion task activated the V5 region. In the control group, the angle task activated the occipitoparietal cortex. Patients and controls showed similar levels of activation, except on the angle task for which occipitoparietal activation was lower in patients than controls. Conclusion: Distinct visuoperceptual functions can be tested in patients with dementia and extrapyramidal motor features when tests use individualized thresholds, adapted tasks, and specialized response systems.

Mesenchymal stem cells and collagen patches for anterior cruciate ligament repair

AIM: To investigate collagen patches seeded with mesenchymal stem cells (MSCs) and/or tenocytes (TCs) with regards to their suitability for anterior cruciate ligament (ACL) repair. METHODS: Dynamic Intraligamentary Stabilization (DIS) utilizes a dynamic screw system to keep ACL remnants in place and promote biological healing, supplemented by collagen patches. How these scaffolds interact with cells and what type of benefit they provide has not yet been investigated in detail. Primary ACL-derived TCs and human bone marrow derived MSCs were seeded onto two different types of 3D collagen scaffolds, Chondro-Gide® (CG) and Novocart® (NC). Cells were seeded onto the scaffolds and cultured for 7 days either as a pure populations or as “premix” containing a 1 : 1 ratio of TCs to MSCs. Additionally, as controls, cells were seeded in monolayers and in co-cultures on both sides of porous high-density membrane inserts (0.4µm). We analyzed the patches by real time polymerase chain reaction (RT-PCR), glycosaminoglycan (GAG), DNA and hydroxy-proline (HYP) content, was determined. To determine cell spreading and adherence in the scaffolds microscopic imaging techniques, i.e. confocal laser scanning microscopy (cLSM) and scanning electron microscopy (SEM), were applied. RESULTS: CLSM and SEM imaging analysis confirmed cell adherence onto scaffolds. The metabolic cell activity revealed that patches promote adherence and proliferation of cells. The most dramatic increase in absolute metabolic cell activity was measured for CG samples seeded with tenocytes or a 1:1 cell premix. Analysis of DNA content and cLSM imaging also indicated MSCs were not proliferating as nicely as tenocytes on CG. The HYP to GAG ratio significantly changed for the premix group, resulting from a slightly lower GAG content, demonstrating that the cells are modifying the underlying matrix. Real-time quantitative polymerase chain reaction data indicated that MSCs showed a trend of differentiation towards a more tenogenic-like phenotype after 7 days. CONCLUSION: CG and NC are both cyto-compatible with primary MSCs and TCs; TCs seemed to perform better on these collagen patches than MSCs.

Asociación entre caracteres biométricos y calidad fisiológica en semillas de tomate (Solanum lycopersicum L.)

La clasificación de las semillas de especies olerícolas se realiza principalmente por peso y tamaño, con criterios similares a los aplicados en cereales y leguminosas, en que se asocia positivamente estos atributos físicos con la calidad fisiológica. No obstante lo anterior, en diversas especies de hortalizas la información es escasa y contradictoria al respecto, lo que motiva la realización de la presente investigación. En semillas de tomate (Solanum lycopersicum L.) se determinó el efecto del peso y tamaño sobre la calidad fisiológica expresada como germinación y vigor. Además, se correlacionaron los resultados de las pruebas de evaluación de calidad fisiológica y se describieron variables del crecimiento y desarrollo. Se utilizaron lotes de diferentes variedades de semillas híbridas de cuatro temporadas, producidas en un clima templado cálido con lluvias invernales y estación seca prolongada (32º 54’ y 34° 21´ latitud Sur). Se midió peso y tamaño de semillas, además en dos temporadas se evaluaron las características internas de área y peso de embrión y área de endospermo. Se determinó la calidad de las semillas con la prueba de germinación y según fuera el año de estudio se midió vigor con las pruebas de envejecimiento acelerado, de plantas útiles al trasplante y de plántulas emergidas. Con análisis de imágenes y rayos X se extrajeron datos del tamaño externo e interno de las semillas y plántulas. Los lotes se compararon mediante análisis de varianza y las medias con la prueba de Tukey, la asociación entre dos variables se determinó con correlaciones de Pearson, las variables de peso y tamaño de la semilla y su relación con las pruebas de calidad, se analizaron mediante regresiones múltiples. Se utilizó un nivel de significación de 0,05 de probabilidad. Los resultados indicaron que el tamaño y no el peso de las semillas de tomate, diferenciaron calidad entre lotes en las diversas variedades. La prueba de germinación tuvo una baja sensibilidad para discriminar lotes, además de una escasa correlación con las características físicas de las semillas, cuando hubo asociación, la relación fue débil y negativa. La prueba de vigor de envejecimiento acelerado diferenció lotes y presentó escasa asociación con las características físicas de las semillas. El número de semillas germinadas en la prueba de envejecimiento acelerado se explicó por el efecto del tamaño de las semillas, mientras que las fracciones de descarte se asociaron con el peso de las mismas. La prueba de vigor de plantas útiles al trasplante no discriminó entre lotes. Tuvo una asociación débil con el peso y tamaño de las semillas. El modelo asociado a esta relación explicó con un alto coeficiente de determinación que el peso de la semilla influyó sobre la emergencia temprana, mientras que la relación fue menor y negativa con plantas de mayor desarrollo. La prueba de vigor de plántulas emergidas discriminó lotes de semillas con plántulas de 3 a 5 días después de siembra. Hubo escasa y débil asociación entre esta prueba y las características de peso y tamaño las semillas. El modelo de predicción de plántulas emergidas fue particular en cada temporada, cuando hubo un coeficiente de determinación alto influyó negativamente el peso o tamaño de la semilla. Entre las pruebas de calidad fisiológica evaluadas en semillas de tomate hubo escasas correlaciones significativas. Entre germinación y vigor las correlaciones significativas fueron débiles y sólo se encontraron en algunas temporadas de evaluación. Entre las pruebas de vigor no hubo asociación. En las pruebas de vigor de plantas útiles al trasplante y de plántulas emergidas, los cotiledones alcanzaron el mayor porcentaje de materia seca y se correlacionaron fuertemente con la materia seca total. En la prueba de plántulas emergidas la materia seca de las radículas diferenció parcialmente lotes de semillas al igual que la longitud total y de las radículas. La longitud de la radícula se correlacionó fuertemente con la longitud total de plántulas. ABSTRACT Seed selection for olericultural species is mainly carried out considering weight and size with similar criteria to those applied in cereals and legumes where size and physiological quality are favorably associated. However, information about several species is limited and contradictory regarding the above, leading to the present research. In tomato (Solanum lycopersicum L.) seeds, the effect of weight and size on the physiological quality expressed as germination and vigor was determined. In addition, results of quality evaluation tests were correlated and variables of growth and development were described. Batches of hybrid seeds from four seasons were used. These seeds were produced in a mild warm climate with winter rainfalls and long dry season (32º 54’ and 34° 21´South Latitude). Seed weight and size were determined, additionally internal characteristics such as embryo area and weight as well as endosperm area were evaluated in two seasons. The quality of seeds was established using the germination test and, depending on the year of the study, vigor was measured through accelerated aging tests for plants useful for transplanting and emerged seedlings. Using imaging analysis and X rays, data regarding external and internal size of seeds and seedlings were obtained. Batches were compared through ANOVA and means using Tukey’s test; the association between both variables was determined with Pearson correlations, whereas variables of seed weight and size and their relation to quality tests were analyzed through multiple regressions. A significance level of 0.05 probability was used. Results showed that the size (but not the weight) of tomatoes differentiates quality between batches from several seasons. The germination test was not sensitive enough to discriminate batches in addition to having a limited correlation with the characteristics of seeds, when they were associated, the relation was weak and unfavorable. Vigor test for accelerated aging made the difference between batches and presented low association with physical characteristics of the seeds. The number of germinated seeds in the accelerated aging test was explained by the effect of the seed size, whereas cull fractions were associated with their weight. The vigor test of plants useful for transplanting did not discriminate between batches. The association with seed weight and size was weak. The model associated to this relation explained, with a high coefficient determination, that the seed weight had influence on early emergence, whereas the relation was minor and unfavorable with more developed plants. Vigor test of emerged seedlings discriminated batches of seeds with seedlings of 3 to 5 days after sowing. There was a limited and weak association between this test and the characteristics of seed weight and size. The prediction model for seedlings emerged was particular in each season, when the determination coefficient was high, seed weight and size influenced negatively. Among the physiological quality tests evaluated in tomato seeds, significant correlations were negligible. Between germination and vigor, significant correlations were poor, being only found in some evaluation seasons. There was no association in the vigor tests. In vigor tests for plants useful for transplanting and emerged seedlings, cotyledons reached the highest percentage of dry matter and were strongly correlated with total dry matter. In the test of emerged seedlings, dry matter of radicles partially differentiated batches of seeds as well as total length and radicles. Radicle length was strongly correlated with total seedlings length.

Auditoria Energética à Escola Secundária Padre Benjamim Salgado - Vila Nova de Famalicão

A dependência energética das grandes economias mundiais alertou o mundo para a necessidade de mudar o comportamento relativo ao consumo de energia. O aumento dos custos energéticos tem induzido ao uso racional da energia por parte das organizações, implicando uma monitorização permanente nas instalações. A eficiência energética é um tema que tem vindo a assumir grande importância nos dias de hoje na nossa sociedade, não só pelos compromissos internacionais assumidos, como também pelo combate ao desperdício energético. Esta dissertação relata o estágio curricular na Agência de Energia do Ave, que inclui uma auditoria energética à escola Padre Benjamim Salgado. O estágio visou essencialmente a contribuição para um modelo de desenvolvimento sustentável, na procura de alternativas com menor impacto ambiental, introduzindo conceitos de eficiência energética e ambiental nos processos de planeamento. Acções de sensibilização e auditorias energéticas foram os projetos que, durante o período curricular, tiveram maior intervenção por parte do autor desta dissertação, tendo em vista a mudança de hábitos de consumo energético na sociedade. A auditoria energética à escola Padre Benjamim Salgado, Joane Vila Nova de Famalicão, apresenta uma abordagem para a determinação da redução dos custos de energia, mantendo o conforto para os seus ocupantes. A escola em análise configura um Grande Edifício de Serviços (GES) à luz de uma análise térmica feita ao edifício escolar, com os valores de eferência atualizados, com entrada em vigor a 31 de dezembro de 2015, ao abrigo da nova regulamentação (decreto-lei n.º118/2013, 20 de Agosto) verificando-se o cumprimento dos requisitos mínimos definidos na Portaria n.º 349/2013 do atual decreto. Foi ainda efetuada, após um levantamento exaustivo, uma análise detalhada das faturas de energia, gás e água, e de todos os equipamentos consumidores de energia que se encontram instalados na instituição.

Surface analysis of lipids by mass spectrometry : More than just imaging

Mass spectrometry is now an indispensable tool for lipid analysis and is arguably the driving force in the renaissance of lipid research. In its various forms, mass spectrometry is uniquely capable of resolving the extensive compositional and structural diversity of lipids in biological systems. Furthermore, it provides the ability to accurately quantify molecular-level changes in lipid populations associated with changes in metabolism and environment; bringing lipid science to the "omics" age. The recent explosion of mass spectrometry-based surface analysis techniques is fuelling further expansion of the lipidomics field. This is evidenced by the numerous papers published on the subject of mass spectrometric imaging of lipids in recent years. While imaging mass spectrometry provides new and exciting possibilities, it is but one of the many opportunities direct surface analysis offers the lipid researcher. In this review we describe the current state-of-the-art in the direct surface analysis of lipids with a focus on tissue sections, intact cells and thin-layer chromatography substrates. The suitability of these different approaches towards analysis of the major lipid classes along with their current and potential applications in the field of lipid analysis are evaluated. © 2013 Elsevier Ltd. All rights reserved.

Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis

Systems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution.

Medical imaging and Biomechanical analysis of scoliosis progression in the growing adolescent spine

Progression of spinal deformity in children was studied with Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) to identify how gravity affects the deformity and to determine the full three-dimensional character of the deformity. The CT study showed that gravity is significant in deformity progression in some patients which has implications for clinical patient management. The world first MRI study showed that the standard clinical measure used to define the extent of the deformity is inadequate and further use of three-dimensional MRI should be considered by spinal surgeons.

Meta-analysis of structural imaging findings in Attention-Deficit/Hyperactivity Disorder

Background Although there are many structural neuroimaging studies of attention-deficit/hyperactivity disorder (ADHD) in children, there are inconsistencies across studies and no consensus regarding which brain regions show the most robust area or volumetric reductions relative to control subjects. Our goal was to statistically analyze structural imaging data via a meta-analysis to help resolve these issues. Methods We searched the MEDLINE and PsycINFO databases through January 2005. Studies must have been written in English, used magnetic resonance imaging, and presented the means and standard deviations of regions assessed. Data were extracted by one of the authors and verified independently by another author. Results Analyses were performed using STATA with metan, metabias, and metainf programs. A meta-analysis including all regions across all studies indicated global reductions for ADHD subjects compared with control subjects, standardized mean difference equal to .408, p less than .001. Regions most frequently assessed and showing the largest differences included cerebellar regions, the splenium of the corpus callosum, total and right cerebral volume, and right caudate. Several frontal regions assessed in only two studies also showed large significant differences. Conclusions This meta-analysis provides a quantitative analysis of neuroanatomical abnormalities in ADHD and information that can be used to guide future studies.

Analysis of focal adhesion turnover: A quantitative live-cell imaging example

Recent advances in optical and fluorescent protein technology have rapidly raised expectations in cell biology, allowing quantitative insights into dynamic intracellular processes like never before. However, quantitative live-cell imaging comes with many challenges including how best to translate dynamic microscopy data into numerical outputs that can be used to make meaningful comparisons rather than relying on representative data sets. Here, we use analysis of focal adhesion turnover dynamics as a straightforward specific example on how to image, measure, and analyze intracellular protein dynamics, but we believe this outlines a thought process and can provide guidance on how to understand dynamic microcopy data of other intracellular structures.