Fractal evaluation in retinographies of individuals with different grades of diabetic retinopathy / Avaliação fractal de retinografias de indivíduos com diferentes graus de retinopatia diabética

ABSTRACT Purpose To evaluate the retinal blood vascular network of the retinographies of patients with different grades of diabetic retinopathy. Methods Ninety Retinographies (MESSIDOR database) were used, with different grades of diabetic retinopathy divided into 4 groups: no retinopathy (n=23), grade one (n=20), grade two (n=20) and grade three (n=27) diabetic retinopathy. The grades of diabetic retinopathy were classified according to the number of microaneurysms, number of hemorrhages and the presence of neovascularization. The images were skeletonized and quantified by fractal methods: dimension of box-counting (Dbc) and information (Dinf). Results The means of Dbc values of groups were around 1.25, without statistically significant difference in the dimension values between groups for whole retina. There was also no statistical difference in Dinf values between groups, whose means ranged between 1.294 ± 0.013 (group of grade 1) and 1.3 ± 0.017 (group of grade 3). The retinographies were divided into regions of equal areas. The fractal values of some retinal regions showed statistical differences, but these differences were not enough to show the sensitivity of fractal methods in identifying diabetic retinopathy. Conclusion The fractal methods were not able to identify the different grades of diabetic retinopathy in retinographies.

RESUMO Objetivo Avaliar a rede vascular sanguínea da retina a partir de retinografias de pacientes com diferentes graus de retinopatia diabética. Métodos Foram utilizadas 90 retinografias (banco de dados MESSIDOR), com diferentes graus de retinopatia diabética divididas em quatro grupos: sem retinopatia (n=23), retinopatia diabética de grau um (n=20), grau dois (n=20) e grau três (n=27). Os graus de retinopatia foram classificados conforme o número de microaneurismas, número de hemorragias e presença de neovascularização. As imagens foram esqueletizadas e quantificadas pelos métodos fractais: dimensão da contagem de caixas e informação. Resultados As médias dos valores das dimensões de contagem de caixas para todos os grupos foram próximas a 1,25, sem diferença estatisticamente significativa nos valores das dimensões entre os grupos para retina inteira. Também não houve diferença estatística nos valores da dimensão de informação entre os grupos, cujas médias variaram entre 1,294 ± 0,013 (grupo do grau 1) e 1,3 ± 0,017 (grupo do grau 3). As imagens retinianas foram divididas em regiões de áreas iguais. Os valores fractais de algumas regiões retinais mostraram diferenças estatísticas, mas estas não foram suficientes para mostrar a sensibilidade dos métodos fractais na identificação da retinopatia diabética. Conclusão Os métodos fractais não foram capazes de identificar os diferentes graus de retinopatia diabética em retinografias.

Can a hypercholesterolemic diet change the basal brain electrical activity and during status epilepticus in rats?

The brain is an organ rich in lipids, including cholesterol, in which these lipids are associated to structure and brain function. Thus alterations in lipid levels of diets may interfere in the brain electrical activity. Our aim was to evaluate the interference of hypercholesterolemic diets in the brain electrical activity in normal individuals and with epilepsy. Histological analysis and electrocorticograms (ECoG) were performed in animals fed with and without hypercholesterolemic diet before and during the status epilepticus induced by pilocarpine. The power spectrum of ECoG was used to estimate the contribution of different brain rhythms in ECoG signal. The animals submitted to the status epilepticus showed cell death, vacuolization with destructuration of the cell layers. Both animal groups, those with status epilepticus and status epilepticus with hypercholesterolemic diet, showed cellular lesions similar. The hyperlipid diet promoted increase of brain electrical activity, this was revealed by increase in the average power of beta wave (14-30 Hz) and decrease in the average power of the delta wave (0,5-4 Hz). This increase of brain electrical activity was even higher when the animals were fed a hypercholesterolemic diet and submitted to status epilepticus. Animals fed with hypercholesterolemic diet and submitted to status epilepticus presented a higher increase in brain excitability compared to control animals. We observed that hypercholesterolemic diet favored a greater severity of the status epilepticus.

Effect of diet with omega-3 in basal brain electrical activity and during status epilepticus in rats.

Western diets are high in saturated fat and low in omega-3. Certain animals cannot produce omega-3 from their own lipids, making it necessary for it to be acquired from the diet. However, omega-3s are important components of the plasma membrane, and altering their proportions can promote physical and chemical alterations in the membranes, which may modify neuronal excitability. These alterations occur in healthy individuals, as well as in patients with epilepsy who are more sensitive to changes in brain electrical activity. This study evaluated the effect of a diet supplemented with omega-3 on the basal brain electrical activity both before and during status epilepticus in rats. To evaluate the brain electrical activity, we recorded electrocorticograms (ECoG) of animals both with and without omega-3 supplementation before and during status epilepticus induced by pilocarpine. Calculation of the average brain wave power by a power spectrum revealed that omega-3 supplementation reduced the average power of the delta wave by 20% and increased the average power of the beta wave by 45%. These effects were exacerbated when status epilepticus was induced in the animals supplemented with omega-3. The animals with and without omega-3 supplementation exhibited increases in basal brain electrical activities during status epilepticus. The two groups showed hyperactivity, but no significant difference between them was noted. Even though the brain activity levels observed during status epilepticus were similar between the two groups, neuron damage to the animals supplemented with omega-3 was more slight, revealing the neuroprotective effect of the omega-3.

Fractal analysis of extra-embryonic vessels of chick embryos under the effect of glucosamine and chondroitin sulfates.

Like heparan sulfate proteoglycans, some monosaccharides and glycosaminoglycans, such as sulfated glucosamine (GS) and chondroitin (CS), integrate the vascular extracellular matrix and may influence vascular endothelial cell growth. To assess the effects of these substances on blood vessel formation, we used the chick yolk sac membrane (YSM) model and fractal geometry quantification, which provided an objective in vivo method for testing potential agents that promote vasculogenesis and angiogenesis. An image processing method was developed to evaluate YSM capillary vessels after they were implanted in a methylcellulose disk of GS or CS at a concentration between 0.001-0.1mg/disk (performed on 2-day old embryos). This method resulted in a binary image of the microvascular network (white vessels on a black background). Fractal box-counting (DBC) and information (DINF) dimensions were used to quantify the activity of GS and CS in vasculogenesis and angiogenesis. YSM treated with GS (0.001-0.1mg) and CS (0.03-0.1mg) showed an increase in fractal dimensions that corresponded to vitelline vessel growth compared to the control group (vehicle), with GS displaying higher fractal dimension values.

Multifractal dimension and lacunarity of yolk sac vasculature after exposure to magnetic field.

Several studies have reported about the effects of magnetic fields (MFs) on vascular tissue. Extremely low frequency magnetic fields (ELF-MFs) can promote either inhibition or stimulation of vasculogenesis and angiogenesis, depending upon the intensity and time of exposure to the MF. To investigate the possible effects of ELF-MF on vascular processes, it is necessary to employ methods that allow parameterization of the vascular network. Vascular network is a structure with fractal geometry; therefore, fractal methods have been used to evaluate its morphometric complexity. Here, we used the lacunarity parameter (complementary method of fractal analysis) and multifractal analyses to investigate angiogenesis and vasculogenesis in the embryonic yolk sac membrane (YSM) of Japanese quails (Coturnix japonica) with and without exposure to an external MF of 1 mT and 60 Hz. Lacunarity results showed that the vascular density was lower for the group exposed to the magnetic field for 9 h/day. In addition, multifractal analysis showed reduced vascularization in the experimental groups (6 h/day and 9 h/day of exposure to MF). Furthermore, multifractal analysis showed difference between the groups exposed for 12 and 24 h/day. Using multifractal methods (generalized dimensions and singularity spectrum), it was possible to characterize the vascular network of the quail embryo YSM as a multifractal object, therefore proving this method to be a more appropriate application than the traditional monofractal methods.