Comparison of the whole slide imaging and conventional light microscopy in the grading of oral epithelial dysplasia: a multi-institutional study.

BACKGROUND: Whole Slide Imaging (WSI) is an alternative method to light microscopy (LM). However, few studies have compared the diagnostic agreement between WSI and LM, especially to grade oral epithelial dysplasia (OED). The purpose of this study was to evaluate the variability in grading OED by the World Health Organization grading system, using WSI and conventional LM, and to investigate whether the access to clinical information, and psychologic or physical states of the pathologists could interfere with the diagnosis. MATERIAL AND METHODS: eleven experienced pathologists from seven Brazilian universities independently evaluated twenty-five OED cases. The analyses were performed in duplicate for each method, with an interval of at least 30 days, and the time consumed in each analysis was measured. Physical and psychologic states were evaluated by blood pressure levels, heart rate and two questionnaires: State-Trait Anxiety Inventory and Perceived Stress Scale. Clinical information was provided after the second evaluation using WSI and the pathologist could change their diagnostic decision or not. RESULTS: LM showed a higher inter-examiner agreement (k=0.53) than WSI (k=0.45) and a smaller time consumed by the pathologists (mean of 65.53 seconds compared to 91.02 seconds in WSI). In the first analysis using conventional microscopy, there was a positive correlation between kappa values and anxiety (r=0.47, p=0.02), and stress (r=0.64, p<0.01), and an inverse correlation with heart rate (r=-0.48, p=0.02). In the digital analysis, there was also a positive correlation between kappa values and anxiety (r=0.75, p<0.001). After clinical information was given, there was a slight change in 11.3% of the cases, and a great discrepancy in 1.1% of the cases, mainly increasing the OED grade. CONCLUSIONS: both microscopy systems had similar results, although LM had slightly higher kappa values, and WSI was more time consuming.

CD4+ T helper cells and regulatory T cells in active lupus nephritis: an imbalance towards a predominant Th1 response?

The objective of this study was to evaluate the frequency of CD4+ T cell subsets in peripheral blood mononuclear cells (PBMC), urine and renal tissue from patients with lupus nephritis (LN). PBMC and urinary cells were collected from 17 patients with active LN, 20 disease controls (DC) with primary glomerulonephritis and 10 healthy controls (HC) and were analysed by flow cytometry with markers for T helper type 1 (Th1), Th2, Th17 and regulatory T cells (Treg ) cells. T cell subsets were assessed by immunohistochemistry from LN biopsy specimens from 12 LN patients. T cell subtypes in PBMC were re-evaluated at 6 months of therapy. CD4+ T cells were decreased in PBMC in LN compared with DC and HC (P = 0·0001). No differences were observed in urinary CD4+ T cell subsets between LN and DC. The frequency of urinary Th17 cells was higher in patients with non-proliferative than in proliferative LN (P = 0·041). CD3+ and T-box 21 ( Tbet+) cells were found in glomeruli and interstitium of LN patients, while forkhead box protein 3 (FoxP3), retinoid-related orphan receptor gamma (ROR-γ) and GATA binding protein 3 (GATA-3) were present only in glomeruli. Th1 cells in PBMC were correlated negatively with urinary Th1 cells (Rho = -0·531; P = 0·028) and with Tbet in renal interstitium (Rho = -0·782; P = 0·004). At 6 months, LN patients showed an increase in Th17 cells in PBMC. In conclusion, the inverse association between Th1 cells from PBMC and urinary/renal tissue indicate a role for Th1 in LN pathophysiology. Urinary Th17 cells were associated with less severe LN, and Th17 increased in PBMC during therapy. Urinary CD4+ T cells were not different between LN and DC.

DNA interaction with DAPI fluorescent dye: Force spectroscopy decouples two different binding modes.

In this work, we use force spectroscopy to investigate the interaction between the DAPI fluorescent dye and the λ-DNA molecule under high (174 mM) and low (34 mM) ionic strengths. Firstly, we have measured the changes on the mechanical properties (persistence and contour lengths) of the DNA-DAPI complexes as a function of the dye concentration in the sample. Then, we use recently developed models in order to connect the behavior of both mechanical properties to the physical chemistry of the interaction. Such analysis has allowed us to identify and to decouple two main binding modes, determining the relevant physicochemical (binding) parameters for each of these modes: minor groove binding, which saturates at very low DAPI concentrations ( CT ∼ 0.50 µM) and presents equilibrium binding constants of the order of ∼107 M-1 for the two ionic strengths studied; and intercalation, which starts to play a significant role only after the saturation of the first mode, presenting much smaller equilibrium binding constants (∼105 M-1 ).

Marandu grass pastures with the same average height and different horizontal structures change the feeding behavior of grazing sheep? / Pastos de capim-marandu com mesma altura média e diferentes estruturas horizontais alteram o comportamento ingestivo de ovinos em pastejo?

The objective was to investigate the feeding behavior of sheep in marandu grass (Brachiaria brizantha cv. Marandu) pastures under continuous stocking with the same average height and different horizontal structures. The treatments were composed of less heterogeneous pastures (P-H, 24% coefficient of variation of plant heights and more heterogeneous pastures (P+H, 46% coefficient of variation of plant heights) and the locations of the picket, front and back, were also evaluated. A randomized block design was used, with split plots in space and four replications in two years. Crossbred lambs were used, managed under continuous stocking and variable stocking rate, to maintain average height of pastures at 30 cm. Greater masses of total forage, live leaf, live stem and dead material, and greater time of sheep in rumination, occurred in the front, in relation to the paddock background. In P-H, grazing time (GT) did not vary between regions. In P+H, GT was lower at the front than at the background. Idle time (IT) was similar between P-H and P+H. However, the animals remained more IT in the front than in the background. The feeding behavior of grazing sheep is modified in marandu grass pastures under continuous stocking with the same average height, but with different spatial variability of the vegetation.

Objetivou-se investigar o comportamento ingestivo de ovinos em pastos de capim-marandu (Brachiaria brizantha cv. Marandu) sob lotação contínua com mesma altura média e diferentes estruturas horizontais. Os tratamentos foram compostos de pastos menos heterogêneo (P-H, 24% de coeficiente de variação das alturas das plantas) e pastos mais heterogêneo (P+H, 46% de coeficiente de variação das alturas das plantas), também foi avaliado os locais do piquete, frente e fundo. O delineamento utilizado foi de blocos ao acaso, com parcelas subdivididas no espaço e quatro repetições em dois anos. Foram utilizadas borregas mestiças, manejadas sob lotação continua e com taxa de lotação variável, para manter altura média dos pastos em 30cm. Maiores massas de forragem total, folha viva, colmo vivo e material morto, e maior tempo dos ovinos em ruminação, ocorreram na frente, em relação ao fundo do piquete. No P-H, o tempo em pastejo (TP) não variou entre as regiões. No P+H, o TP foi menor na frente do que no fundo. O tempo em ócio (TO) foi semelhante entre os P-H e P+H. Porém, os animais permaneceram mais TO na frente que no fundo. O comportamento ingestivo de ovinos em pastejo é modificado em pastos de capim-marandu sob lotação contínua com mesma altura média, porém com diferentes variabilidades espaciais da vegetação.

Chemical composition of Andropogon gayanus cv. planaltina predicted through nirs and analyzed through wet chemistry / Composição químico-bromatológica de Andropogon gayanus, cultivar Planaltina, predita pelo NIRS e analisada por via úmida

Objetivou-se comparar os teores dos nutrientes do capim Andropogon gayanus obtidos no equipamento NIRS com os valores obtidos por análises de via úmida, para identificar se os modelos globais de calibração do equipamento NIRS são adequados para predizer a composição químico-bromatológica. As análises de via seca no NIRS foram executadas no Laboratório de Bromatologia e Nutrição Animal (Laban), pertencente à Faculdade de Medicina Veterinária (Famev) da Universidade Federal de Uberlândia (UFU), enquanto as análises de via úmida foram realizadas no Laboratório de Química Analítica de Plantas da Embrapa Cerrados, localizado em Brasília-DF. As amostras utilizadas foram de capim Andropogon gayanus, cultivar Planaltina, as quais, após corte de 20cm acima do solo, foram secas e moídas no tamanho de 1mm. Após a moagem, foram analisadas por via úmida para os teores de matéria seca (MS), matéria mineral (MM), proteína bruta (PB), fibra insolúvel em detergente neutro (FDN) e fibra insolúvel em detergente ácido (FDA), segundo metodologias propostas por Detmann et al. (2012). As amostras moídas foram colocadas em cubetas próprias do equipamento e escaneadas em espectrômetro de refletância no infravermelho próximo (NIRS) NIR, modelo Spectra Star 2600 XT series of Near Infrared Analyzers (Unity Scientific®), em duplicata. Para obtenção dos teores de MS, MM, PB, FDN e FDA, empregaram-se modelos de calibração do próprio equipamento NIRS, utilizando-se a curva de calibração própria do equipamento. As médias dos teores de MS, MM, PB, FDN e FDA obtidas pelas análises de bancada (via úmida) e pelo uso do equipamento NIRS foram comparadas pelo teste T (dados pareados), ao nível de 5% de probabilidade, empregando-se o delineamento em blocos ao acaso. Houve diferença significativa nos nutrientes preditos pelo NIRS e analisados por via úmida (P≤0,05). O NIRS superestimou os teores de matéria seca e matéria mineral, mas subestimou os teores de FDN, FDA e PB. Esse resultado mostra que, quando do uso de modelos globais de calibração, esse equipamento não é capaz de predizer corretamente a composição químico-bromatológica de forrageiras de clima tropical.

Nutritional and metabolic parameters of sheep on deferred pasture with different strategy changes during the dry season / [Parâmetros nutricionais e metabólicos de ovinos em pasto diferido com diferentes estratégias mudam ao longo do período seco]

To verify if strategies to reduce the height of Marandu grass pasture at beginning of the deferral period change intake and metabolic parameters in sheep during the dry season, 18 crossbred females, ½ Dorper + ½ Santa Inês were used, distributed in 9 Marandu grass paddock handles to 3 drawdown strategies at the beginning of deferral period: 1) maintenance of grass with 15cm for 5 months before deferral start (15/15cm); 2) maintenance of marandu grass with 25 cm for 5 months, but at the beginning of deferral it was reduced to 15cm (25/15cm); 3) maintenance of 35 cm grass for 5 months, but at the beginning of deferral it was reduced to 15cm (35/15cm). The experiment was conducted in a completely randomized design (CRD), with 6 repetitions and repeated measures over time. Strategies to reduce the height of Marandu grass pasture, at the beginning of the deferral period, do not change nutrient intake and energy and protein metabolism. However, changes occurring in the pasture during the dry season alter these variables, and deferral is an interesting strategy to be used for forage production in winter, however, to optimize its use, it is necessary to use supplementation with concentrated rich in fermentable carbohydrates.

Para verificar se estratégias de redução da altura do pasto de capim-marandu, no início do período de diferimento, alteram consumo e parâmetros metabólicos em ovinos durante o período seco, foram utilizadas 18 fêmeas, mestiças, ½ Dorper + ½ Santa Inês, que foram distribuídas em nove piquetes de capim-marandu, com três estratégias de rebaixamento no início do período de diferimento: 1) manutenção do capim com 15cm, por cinco meses, antes do início do diferimento (15/15cm); 2) manutenção do capim-marandu com 25cm, por cinco meses, rebaixado, no início do diferimento, para 15cm (25/15cm); e 3) manutenção do capim com 35cm durante cinco meses, rebaixado, no início do diferimento, para 15cm (35/15cm). O experimento foi conduzido em delineamento inteiramente ao acaso (DIC), com seis repetições e medidas repetidas no tempo. Estratégias de redução da altura do pasto, no início do período de diferimento, não alteram o consumo de nutrientes e metabolismo energético e proteico. Porém, as mudanças no pasto durante o período seco alteram essas variáveis, sendo o diferimento uma estratégia interessante para utilizar na produção de forragem no inverno. Para otimizar seu uso, faz-se necessária, entretanto, a utilização da suplementação com concentrados ricos em carboidratos rapidamente fermentáveis.

Extracellular vesicles: structure, function, and potential clinical uses in renal diseases.

Interest in the role of extracellular vesicles in various diseases including cancer has been increasing. Extracellular vesicles include microvesicles, exosomes, apoptotic bodies, and argosomes, and are classified by size, content, synthesis, and function. Currently, the best characterized are exosomes and microvesicles. Exosomes are small vesicles (40-100 nm) involved in intercellular communication regardless of the distance between them. They are found in various biological fluids such as plasma, serum, and breast milk, and are formed from multivesicular bodies through the inward budding of the endosome membrane. Microvesicles are 100-1000 nm vesicles released from the cell by the outward budding of the plasma membrane. The therapeutic potential of extracellular vesicles is very broad, with applications including a route of drug delivery and as biomarkers for diagnosis. Extracellular vesicles extracted from stem cells may be used for treatment of many diseases including kidney diseases. This review highlights mechanisms of synthesis and function, and the potential uses of well-characterized extracellular vesicles, mainly exosomes, with a special focus on renal functions and diseases.

DNA interaction with diaminobenzidine studied with optical tweezers and dynamic light scattering.

We have studied the interaction of the DNA molecule with the ligand 3,3'-diaminobenzidine (DAB) by performing single molecule stretching experiments with optical tweezers and dynamic light scattering (DLS) on the DNA-DAB complexes. In the stretching experiments, the persistence and contour lengths of the complexes were measured as a function of DAB concentration, allowing one to infer the main binding mechanism and also to determine the physicochemical parameters of the interaction. In the DLS experiments, the effective size of the complexes, measured as the hydrodynamic radius, was monitored as a function of DAB concentration. We found a qualitative agreement between the results obtained from the two techniques by comparing the behaviors of the hydrodynamics radius and the radius of gyration, since this last one can be expressed as a function of the persistence and contour lengths.

Extracellular vesicles: structure, function, and potential clinical uses in renal diseases

Interest in the role of extracellular vesicles in various diseases including cancer has been increasing. Extracellular vesicles include microvesicles, exosomes, apoptotic bodies, and argosomes, and are classified by size, content, synthesis, and function. Currently, the best characterized are exosomes and microvesicles. Exosomes are small vesicles (40-100 nm) involved in intercellular communication regardless of the distance between them. They are found in various biological fluids such as plasma, serum, and breast milk, and are formed from multivesicular bodies through the inward budding of the endosome membrane. Microvesicles are 100-1000 nm vesicles released from the cell by the outward budding of the plasma membrane. The therapeutic potential of extracellular vesicles is very broad, with applications including a route of drug delivery and as biomarkers for diagnosis. Extracellular vesicles extracted from stem cells may be used for treatment of many diseases including kidney diseases. This review highlights mechanisms of synthesis and function, and the potential uses of well-characterized extracellular vesicles, mainly exosomes, with a special focus on renal functions and diseases.