Estado nutricional em nitrogênio da grama esmeralda avaliado por meio do teor foliar, clorofilômetro e imagem digital, em área adubada com lodo de esgoto

A intensidade da cor verde da folha pode ser alternativa para estimar a concentração de N na planta, devido à relação entre o teor de clorofila e o de N no tecido foliar. Objetivou-se neste trabalho avaliar índices da cor verde da grama esmeralda obtidos da análise da imagem digital e pelo uso do clorofilômetro para predizer o estado nutricional em N fornecido pelo lodo de esgoto. O experimento foi instalado e desenvolvido em uma propriedade comercial de grama esmeralda, localizada na cidade de Itapetininga (SP). O delineamento experimental foi em blocos casualizados, com quatro repetições e cinco doses de lodo de esgoto: 0, 10, 20, 30 e 40 Mg ha-1, base seca. As doses de lodo aplicadas correspondem a 100, 200, 300 e 400 kg ha-1 de nitrogênio disponível. Foram avaliadas as concentrações de N e a intensidade de coloração verde da folha pelo uso do clorofilômetro (ICV) e por meio da análise da imagem digital (G, H e ICVE) aos 45, 105 e 165 dias após a aplicação do lodo. Os valores de intensidade obtidos foram correlacionados com a concentração de N na lâmina foliar e com a taxa de cobertura do solo determinada nas mesmas épocas. A aplicação de doses de lodo de esgoto proporcionou aumento dos índices de cor verde e da concentração de N nas folhas da grama esmeralda. A concentração de N na lâmina foliar pode auxiliar a adubação nitrogenada em cobertura, pois proporcionou altas correlações com a taxa de cobertura do solo. O matiz (H) obtido com a imagem digital e a intensidade de cor verde da folha (ICV) obtida com o clorofilômetro correlacionaram-se com a concentração de N e com a taxa de cobertura do solo e, dessa forma, podem servir como índices na recomendação da adubação nitrogenada.

Úlceras crônicas dos membros inferiores: avaliação pela fotografia digital

OBJETIVO: Avaliar o desempenho da análise de imagem digital na estimativa da área acometida pelas úlceras crônicas dos membros inferiores. MÉTODOS: Estudo prospectivo em que foram mensuradas úlceras empregando o método planimétrico clássico, utilizando desenho dos seus contornos em filme plástico transparente, medida sua área posteriormente por folha milimetrada. Esses valores foram utilizados como padrão para a comparação com a estimativa de área pelas fotografias digitais padronizadas das úlceras e dos desenhos das mesmas em filme plástico. Para criar um referencial de conversão dos pixels em milímetros, foi empregado um adesivo com tamanho conhecido, adjacente à úlcera. RESULTADOS: foram avaliadas 42 lesões em 20 pacientes portadores de úlceras crônicas de membros inferiores. As áreas das úlceras variaram de 0,24 a 101,65cm². Observou-se forte correlação entre as medidas planimétricas e as fotos das úlceras (R²=0,86 p<0,01), porém a correlação das medidas planimétricas com as fotos digitais dos desenhos das úlceras foi ainda maior (R²=0,99 p<0,01). CONCLUSÃO: A fotografia digital padronizada revelou-se método rápido, preciso e não-invasivo capaz de estimar a área afetada por úlceras. A avaliação das medidas fotográficas dos contornos das úlceras deve ser preferida em relação à análise de sua fotografia direta.

Ischemia and reperfusion in skin flaps: effects of mannitol and vitamin C in reducing necrosis area in a rat experimental model

OBJETIVO: Neste trabalho foi padronizado modelo experimental de isquemia e reperfusão em retalho cutâneo em ratos no qual estudou-se possibilidade de uma solução antioxidante, composta por Ringer lactato, vitamina C e manitol de reduzir a área de necrose. MÉTODOS: O modelo consistiu de levantamento de retalho cutâneo axial de 6,0 x 3,0cm, submetido à isquemia de 8 horas e reperfusão de 7 dias. Os animais foram divididos em quatro grupos: grupos S1, S2 (10 animais cada), C e T (14 animais cada). Nos grupos S1 e S2 todos os procedimentos dos demais grupos foram efetuados, exceto a isquemia e reperfusão: S1 recebeu apenas Ringer lactato e S2 a solução antioxidante. Os grupos C e T foram submetidos à isquemia. O grupo C recebeu somente Ringer lactato e o grupo T a solução antioxidante. No 7(0) dia de pós-operatório as áreas de necrose e pele viável do retalho foram delineadas em decalque de acetato, os quais foram por sua vez analisados em sistema computadorizado KS-300 (Carl Zeiss). RESULTADOS: A análise estatística mostrou que não houve diferenças significativas entre o grupo tratado e controle quanto à área de necrose. CONCLUSÃO: Concluiu-se que o modelo experimental é consistente, determinando área de necrose limitada e uniforme nos animais não tratados e que as drogas usadas, nessa posologia e modo de aplicação, não foram efetivas em diminuir a área de necrose no modelo experimental em questão.

Long term evaluation of morphometric and ultrastructural changes of testes of alloxan-induced diabetic rats

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

Relief grating induced by photo-expansion in Ga-Ge-S and Ga-Ge-As-S glasses

We report the fabrication of relief diffraction gratings recorded on a surface of photosensitive Ga10Ge25S65 and Ga5Ge25As5S65 glasses by means of interference of two UV laser beams at 351 nm. The diffraction efficiency (eta) of first diffraction order was measured. Atomic-force-microscope (AFM) was used to perform a 3D imaging analysis of the sample surface topography that shows the superposition of an imprinted grating over the topography of the glass. The change in the absorption edge and the refractive index has been evaluated and a structural approach of the relief grating on the glass surface has been discussed.

Potencial Fisiológico de sementes de Crotalaria juncea L

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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

Testes de vigor em sementes de alface

O vigor da semente tem sido avaliado por meio de testes fisiológicos, bioquímicos e de tolerância a estresse. Mais recentemente, com o uso da análise computadorizada de imagens de plântulas, podem ser obtidas informações objetivas, em período relativamente curto, com menor interferência humana. O objetivo nesta pesquisa foi identificar testes eficientes para avaliação do vigor de sementes de alface, com ênfase para a análise de imagens digitais de plântulas. Sementes de 12 lotes (seis da cultivar Vanda e seis da Vera) foram avaliadas por meio de testes de germinação, germinação a baixa temperatura (15ºC), emergência de plântulas e envelhecimento acelerado tradicional e com solução salina (NaCl 40%) saturada, a 41ºC/48 h e 41ºC/72 h. Utilizou-se, também, o sistema de análise computadorizada de imagens de plântulas (SVIS®), desenvolvido pela Ohio State University (EUA), que forneceu dados referentes ao índice de vigor, uniformidade e crescimento de plântulas. Os testes foram realizados a cada três meses, durante 12 meses de armazenamento, sob condições ambiente. Com base nos resultados obtidos, concluiu-se que os testes de envelhecimento acelerado com uso de solução saturada de NaCl, a 41ºC por 48 e 72 horas e de velocidade de germinação, e os parâmetros obtidos com o software SVIS, índice de crescimento de plântulas e comprimento da raiz primária, são eficientes na avaliação do vigor de lotes de sementes de alface.

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.

Fine Mapping of qroot-yield-1.06, a QTL for Root, Plant Vigor and Yield in Maize

Root-yield-1.06 is a major QTL affecting root system architecture (RSA) and other agronomic traits in maize. The effect of this QTL has been evaluated with the development of near isogenic lines (NILs) differing at the QTL position. The objective of this study was to fine map qroot-yield-1.06 by marker-assisted searching for chromosome recombinants in the QTL interval and concurrent root phenotyping in both controlled and field conditions, through successive generations. Complementary approaches such as QTL meta-analysis and RNA-seq were deployed in order to help prioritizing candidate genes within the QTL target region. Using a selected group of genotypes, field based root analysis by ‘shovelomics’ enabled to accurately collect RSA information of adult maize plants. Shovelomics combined with software-assisted root imaging analysis proved to be an informative and relatively highly automated phenotyping protocol. A QTL interval mapping was conducted using a segregating population at the seedling stage grown in controlled environment. Results enabled to narrow down the QTL interval and to identify new polymorphic markers for MAS in field experiments. A collection of homozygous recombinant NILs was developed by screening segregating populations with markers flanking qroot-yield-1.06. A first set of lines from this collection was phenotyped based on the adapted shovelomics protocol. QTL analysis based on these data highlighted an interval of 1.3 Mb as completely linked with the target QTL but, a larger safer interval of 4.1 Mb was selected for further investigations. QTL meta-analysis allows to synthetize information on root QTLs and two mQTLs were identified in the qroot-yield-1.06 interval. Trascriptomics analysis based on RNA-seq data of the two contrasting QTL-NILs, confirmed alternative haplotypes at chromosome bin 1.06. qroot-yield-1.06 has now been delimited to a 4.1-Mb interval, and thanks to the availability of additional untested homozygous recombinant NILs, the potentially achievable mapping resolution at qroot-yield-1.06 is c. 50 kb.

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.