Biomechanical and biochemical investigation of erythrocytes in late stage human leptospirosis

Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira, which can cause lipid changes in the erythrocyte membrane. Optical tweezers were used to characterize rheological changes in erythrocytes from patients with leptospirosis in the late stage. Biochemical methods were also used for quantification of plasma lipid, erythrocyte membrane lipid, and evaluation of liver function. Our data showed that the mean elastic constant of erythrocytes from patients with leptospirosis was around 67% higher than the control (healthy individuals), indicating that patient's erythrocytes were less elastic. In individuals with leptospirosis, several alterations in relation to control were observed in the plasma lipids, however, in the erythrocyte membrane, only phosphatidylcholine showed a significant difference compared to control, increasing around 41%. With respect to the evaluation of liver function of individuals with leptospirosis, there was a significant increase in levels of alanine transaminase (154%) and aspartate transaminase (150%), whereas albumin was 43.8% lower than control (P<0.01). The lecithin-cholesterol acyltransferase fractional activity was 3.6 times lower in individuals with leptospirosis than in the healthy individuals (P<0.01). The decrease of the erythrocyte elasticity may be related to the changes of erythrocyte membrane phospholipids composition caused by disturbances that occur during human leptospirosis, with phosphatidylcholine being a strong candidate in the erythrocyte rheological changes.

Long-term correlation of the electrocorticogram as a bioindicator of brain exposure to ionizing radiation

Understanding the effects of radiation and its possible influence on the nervous system are of great clinical interest. However, there have been few electrophysiological studies on brain activity after exposure to ionizing radiation (IR). A new methodological approach regarding the assessment of the possible effects of IR on brain activity is the use of linear and nonlinear mathematical methods in the analysis of complex time series, such as brain oscillations measured using the electrocorticogram (ECoG). The objective of this study was to use linear and nonlinear mathematical methods as biomarkers of gamma radiation regarding cortical electrical activity. Adult Wistar rats were divided into 3 groups: 1 control and 2 irradiated groups, evaluated at 24 h (IR24) and 90 days (IR90) after exposure to 18 Gy of gamma radiation from a cobalt-60 radiotherapy source. The ECoG was analyzed using power spectrum methods for the calculation of the power of delta, theta, alpha and beta rhythms and by means of the α-exponent of the detrended fluctuation analysis (DFA). Using both mathematical methods it was possible to identify changes in the ECoG, and to identify significant changes in the pattern of the recording at 24 h after irradiation. Some of these changes were persistent at 90 days after exposure to IR. In particular, the theta wave using the two methods showed higher sensitivity than other waves, suggesting that it is a possible biomarker of exposure to IR.

Crescimento vascular em membrana do saco vitelínico e desenvolvimento embrionário de codornas japonesas (Coturnix japonica) expostas a campo magnético de baixa frequência / Vascular growth in yolk sac membrane and embryonic development of Japanese quail (Coturnix japonica) exposed to low frequency magnetic field

O objetivo deste trabalho foi observar a influência do campo magnético (CM) de baixa frequência na membrana do saco vitelínico (MSV) e no desenvolvimento do embrião de codornas japonesas (Coturnix japonica) em 72 horas de incubação. Ovos fertilizados foram expostos a nove horas consecutivas de CM, sendo um grupo a partir das 48 horas e o outro a partir das 63 horas de incubação. A quantificação da vascularização da MSV foi determinada pela obtenção da dimensão fractal por meio dos métodos de box-counting e de dimensão de informação, enquanto o peso corporal e o percentual de comprimento cefálico dos embriões foram utilizados como parâmetros de desenvolvimento embrionário. O CM não causou diferenças significativas na densidade vascular da MSV nem no desenvolvimento embrionário, quando comparados ao grupo controle...

The aim of this study was to observe the influence of the low frequency magnetic field (MF) on the yolk sac membrane (YSM) and embryonic development of Japanese quail (Coturnix japonica) in 72 hours of incubation. Fertilized eggs were exposed to 9 consecutive hours of MF, with a group from 48 hours and the second group from 63 hours of incubation. The evaluation of YSM vascularization was determined by the fractal dimensions obtained through box-counting method and information dimension, while body weight of the embryo and percentage of cephalic length were used as parameters for embryo development. The MF caused no significant differences in vessel density in the YSM, nor in the embryonic development considering the body weight and percentage cephalic length, when were compared to the control group...

Efeito dos ácidos graxos essenciais sobre lipidemia e vascularização da membrana vitelina de codornas japonesas / Effect of essential fatty acids on lipidemia and vitelline membrane vascularization in Japanese quails

Verificou-se o efeito de diferentes fontes de ácidos graxos essenciais ômega-3 e ômega-6 sobre o perfil lipídico de codornas japonesas e sobre o crescimento vascular na membrana vitelina dos embriões de aves suplementadas com óleo de peixe, 2% e 4%, e óleo de soja, 2% e 4%, em relação à dieta-controle, sem suplementação lipídica. Foi usado o método enzimático com reação colorimétrica para estimar o perfil lipídico sérico de colesterol total, triglicerídeos e HDL das aves. A vascularização na membrana vitelina foi quantificada por meio da dimensão fractal, utilizando-se o método de box-counting. A concentração de colesterol dos grupos controle e tratados não diferiu entre si. Para o HDL, o grupo que recebeu maior proporção de óleo de peixe, 4%, diferiu dos outros grupos. Os níveis de triglicerídeos das codornas suplementadas com óleo de peixe foram superiores aos dos demais grupos. A dimensão fractal da vascularização da membrana vitelina dos embriões de codornas dos grupos que receberam 4% de óleo de peixe e 4% de óleo de soja foi significativamente menor que a dos demais grupos, indicando efeitos antiangiogênicos no processo de formação vascular.

The effect of different sources of omega-3 and omega-6 essential fatty acids on serum lipid profile in Japanese quails and on vascular growth in the embryos vitelline membrane was verified. The birds were supplemented with fish oil, 2% and 4%, and soybean oil, 2% and 4%, and one control diet, without lipid supply. The enzymatic method with colorimetric reaction was used to estimate the profile of serum total cholesterol, triglycerides and HDL of the birds. Vitelline membrane vascularization was measured using the box-counting fractal dimension. The cholesterol concentration of the treated and control groups showed no difference. For HDL, the group treated with a higher proportion of fish oil, 4%, differed from other groups. The triglycerides levels of quails supplemented with fish oil were higher than the other groups. The fractal dimension of quail embryos vitelline membrane from the groups treated with 4% fish oil and 4% soybean oil was lower than the other groups, indicating antiangiogenic effects on the vascular formation process.

Can the fractal dimension be applied for the early diagnosis of non-proliferative diabetic retinopathy?

The fractal dimension has been employed as a useful parameter in the diagnosis of retinal disease. Avakian et al. (Curr Eye Res 2002; 24: 274-280), comparing the vascular pattern of normal patients with mild to moderate non-proliferative diabetic retinopathy (NPDR), found a significant difference between them only in the macular region. This significant difference in the box-counting fractal dimension of the macular region between normal and mild NPDR patients has been proposed as a method of precocious diagnosis of NPDR. The aim of the present study was to determine if fractal dimensions can really be used as a parameter for the early diagnosis of NPDR. Box-counting and information fractal dimensions were used to parameterize the vascular pattern of the human retina. The two methods were applied to the whole retina and to nine anatomical regions of the retina in 5 individuals with mild NPDR and in 28 diabetic but opthalmically normal individuals (controls), with age between 31 and 86 years. All images of retina were obtained from the Digital Retinal Images for Vessel Extraction (DRIVE) database. The results showed that the fractal dimension parameter was not sensitive enough to be of use for an early diagnosis of NPDR.

Diameter of the mammalian porin channel in open and "closed" states: direct measurement at the single channel level in planar lipid bilayer

Porin isolated from bovine skeletal muscle was reconstituted in planar lipid bilayers under voltage clamp conditions. A set of non-electrolytes were used as molecular probes for determining the pore diameter. The maximal diameter of the open channel was estimated to be 3.02 + 0.26 nm. As observed for other porin channels, a large transmembrane potential drove the channel into a "closed" state. The channel transition to the low conductance (closed) state was followed by a decrease in the maximal diameter of the channel to 2.4 +- 0.08 nm.

The diameter of water pores formed by colicin la in planar lipid bilayers

The effective size of colicin Ia channel was tested by a recently described method 9FEMS, Microbiology and Immunology (1992). 105: 93-100) in which the nonelectrolyte molecules with different hydrodynamic diameters (0.52 to 5.0nm) were used as molecular tools. We have shown that despite low conductance (55-105 pS at 1.5 MKCl, pH 7.0) the ion channels formed by colicin Ia have a fairly large water pore diameter equal to 1.66-1 1.88nm. The results are discussed in terms of an energetic barrier for ions passing into the channel lumen

Hurst analysis in the study of ion channel kinetics

Ion channels are protein molecules which can assume distinct open and closed conformational states. The transitions between these states can be controlled by the electrical field, ions and/or drugs. Records of unitary current events show that short open-time intervals are frequently adjacent to much longer closed-time intervals, and vice-versa, suggesting that the kinetic process has memory, i.e., the intervals are correlated in time. here the rescaled range analysis (R/S Hurst analysis) is proposed as a method to test for correlation. Simulations were performed with a two-state Markovian model, which has no memory. The calculated Hurst coefficients (H) presented a mean + or - SD value of 0.493 + or - 0.025 (N = 100). For the Ca2+ -activated K+ channels of Leydig cells, H wass equal to 0.75, statistically different (1 percent level) from that calculated for the memoryless proces. Randomly shuffling the experimental data resulted in an H = 0.55, not significantly different (1 percent level) from that found for the two-state Markovian model. For a linear three-state Markovian model, H was equal to 0.548 + or - 0.017 (N = 15), agin not significantly different (1 percent level) from that of the memoryless proces. Although the tree-state Markovian model adequately describes the open-and closed-time distributions, it does not account for the correlation found in this Ca2+ -activatedK+ channel. Our results ilustrate the efficacy of the R/S analysis in determining whether successive opening and closing events are correlated in time and can be of help in deciding which odel should be used to describe the kinetics of ion channels

Antigen F1 from yersinia pestis forms aqueous channels in lipid bilayer membranes

Antigen F1 is a protein of 17 kDa produced by Yersinia pestis it is cultured at 37 grade C. When incorporated into planar lipid bilayer membranes this protein induces fluctuations on membrane conductance typical of the formation of ionic channels. These fluctuations reveal two distinct unitary conductance sizes, one in the range of 800 to 1400 pS and the other in the range of 140 to 600 pS. Zero current potential measaurements in the presence of a salt gradient show that the channell is not significantly ion selective. The reversal potential measured in the presence of 0.5 MKCl on the cis side and 0.1 MKCl on the trans side was 3.58 ñ 3.98 mV (N=7). The non-selectivity of the channel, in addition to its large conductance, suggests that it forms large aqueous pores. The present results, taken together with other data showing that nantigen F1 inhibits the activity of phagocytic cells, suggest that antigen F1 acts by forming aqueous pores in the membrane of these target cells