Structural characterisation of bisintercalation in higher-order DNA at a junction-like quadruplex

We report the single-crystal X-ray structure for the complex of the bisacridine bis-(9-aminooctyl(2-(dimethylaminoethyl)acridine-4-carboxamide)) with the oligonucleotide d(CGTACG)2 to a resolution of 2.4 Å. Solution studies with closed circular DNA show this compound to be a bisintercalating threading agent, but so far we have no crystallographic or NMR structural data conforming to the model of contiguous intercalation within the same duplex. Here, with the hexameric duplex d(CGTACG), the DNA is observed to undergo a terminal cytosine base exchange to yield an unusual guanine quadruplex intercalation site through which the bisacridine threads its octamethylene linker to fuse two DNA duplexes. The 4-carboxamide side-chains form anchoring hydrogen-bonding interactions with guanine O6 atoms on each side of the quadruplex. This higher-order DNA structure provides insight into an unexpected property of bisintercalating threading agents, and suggests the idea of targeting such compounds specifically at four-way DNA junctions.

Quantitative single-molecule localization microscopy combined with rule-based modeling reveals ligand-induced TNF-R1 reorganization toward higher-order oligomers

We report on the assembly of tumor necrosis factor receptor 1 (TNF-R1) prior to ligand activation and its ligand-induced reorganization at the cell membrane. We apply single-molecule localization microscopy to obtain quantitative information on receptor cluster sizes and copy numbers. Our data suggest a dimeric pre-assembly of TNF-R1, as well as receptor reorganization toward higher oligomeric states with stable populations comprising three to six TNF-R1. Our experimental results directly serve as input parameters for computational modeling of the ligand-receptor interaction. Simulations corroborate the experimental finding of higher-order oligomeric states. This work is a first demonstration how quantitative, super-resolution and advanced microscopy can be used for systems biology approaches at the single-molecule and single-cell level.

Brainstem oxytocinergic modulation of heart rate control in rats: effects of hypertension and exercise training

Oxytocinergic brainstem projections participate in the autonomic control of the circulation. We investigated the effects of hypertension and training on cardiovascular parameters after oxytocin (OT) receptor blockade within the nucleus tractus solitarii (NTS) and NTS OT and OT receptor expression. Male spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats were trained (55% of maximal exercise capacity) or kept sedentary for 3 months and chronically instrumented (NTS and arterial cannulae). Mean arterial blood pressure (MAP) and heart rate (HR) were measured at rest and during an acute bout of exercise after NTS pretreatment with vehicle or OT antagonist (20 pmol of OT antagonist (200 nl of vehicle)-1). Oxytocin and OT receptor were quantified (35S-oligonucleotide probes, in situ hybridization) in other groups of rats. The SHR exhibited high MAP and HR (P < 0.05). Exercise training improved treadmill performance and reduced basal HR (on average -11%) in both groups, but did not change basal MAP. Blockade of NTS OT receptor increased exercise tachycardia only in trained groups, with a larger effect on trained WKY rats (+31 +/- 9 versus +12 +/- 3 beats min-1 in the trained SHR). Hypertension specifically reduced NTS OT receptor mRNA density (-46% versus sedentary WKY rats, P < 0.05); training did not change OT receptor density, but significantly increased OT mRNA expression (+2.5-fold in trained WKY rats and +15% in trained SHR). Concurrent hypertension- and training-induced plastic (peptide/receptor changes) and functional adjustments (HR changes) of oxytocinergic control support both the elevated basal HR in the SHR group and the slowing of the heart rate (rest and exercise) observed in trained WKY rats and SHR.

Pseudolikelihood equations for Potts MRF model parameter estimation on higher order neighborhood systems

This letter presents pseudolikelihood equations for the estimation of the Potts Markov random field model parameter on higher order neighborhood systems. The derived equation for second-order systems is a significantly reduced version of a recent result found in the literature (from 67 to 22 terms). Also, with the proposed method, a completely original equation for Potts model parameter estimation in third-order systems was obtained. These equations allow the modeling of less restrictive contextual systems for a large number of applications in a computationally feasible way. Experiments with both simulated and real remote sensing images provided good results.

Terapia de reposição hormonal não altera a variabilidade da frequência cardíaca em mulheres pós-menopáusicas

INTRODUÇÃO. Mulheres pós-menopáusicas apresentam maior risco de desenvolvimento de doença arterial coronariana. Estudos observacionais demonstraram que a terapia de reposição hormonal produz efeitos benéficos no perfil lipídico e na modulação autonômica cardíaca. O aumento da variabilidade da freqüência cardíaca (VFC), até então atribuído à reposição hormonal, não foi testado em estudos randomizados, placebo-controlados, delineados para permitir a comparação entre as duas formas mais utilizadas de reposição hormonal. A VFC de 24 horas calculada pelo método não linear Mapa de Retorno Tridimensional permite avaliar tanto a modulação vagal como a simpática. OBJETIVOS Avaliar a modulação autonômica cardíaca de mulheres pósmenopáusicas através da análise da VFC no domínio do tempo e dos índices do Mapa de Retorno Tridimensional no ECG de 24 horas. Testar a hipótese de que a reposição hormonal contínua, seja com estradiol isolado (TRE), seja com estradiol associado à noretisterona (TRH), por um período de três meses, aumenta a VFC nessas mulheres. MÉTODOS Quarenta mulheres pós-menopáusicas (46 a 63 anos; média = 54,6 ± 4,2) foram randomizadas para um dos três tratamentos, de forma contínua: TRH, estrogenioterapia (TRE) ou placebo, por três meses consecutivos. Previamente, todas as mulheres foram submetidas a exames clínico, ginecológico e laboratorial (glicose, estradiol, HDL, LDL, triglicerídios; mamografia e ultrassonografia transvaginal). O ECG de 24 horas foi gravado em cada paciente, antes e após o tratamento, para calcular os índices da VFC. RESULTADOS Não houve diferença estatisticamente significativa entre os três grupos, após 3 meses de tratamento, nos índices da VFC e do Mapa de Retorno Tridimensional. A TRH diferiu da TRE apenas quanto ao perfil lipídico. A associação com a noretisterona provocou uma redução de 12,4 % no HDL (p = 0,008). CONCLUSÃO Em mulheres pós-menopáusicas, a terapia de reposição hormonal contínua com estradiol, ou com estradiol associado à noretisterona, por um período de 3 meses, não altera a modulação autonômica cardíaca avaliada pela VFC.

A Higher order and stable method for the numerical integration of Random Differential Equations

Trabalho apresentado no Congresso Nacional de Matemática Aplicada à Indústria, 18 a 21 de novembro de 2014, Caldas Novas - Goiás

Holter electrocardiography in dogs showing doxorubicin-induced dilated cardiomyopathy

O estudo consistiu na identificação precoce da ocorrência de arritmias em cães com cardiomiopatia dilatada experimental induzida pela doxorrubicina (DOX). Utilizaram-se 10 cães adultos, sadios, distribuídos nos grupos A (n=5) e B (n=5). O grupo B recebeu 30mg/m² de DOX, via intravenosa, a cada 21 dias, até a dose cumulativa de 180 ou 240mg/m². No grupo A (controle), administrou-se solução salina 0,9%, via intravenosa, nos mesmos intervalos do grupo B. Ao se evidenciar o quadro de disfunção miocárdica nos cães do grupo B, caracterizado pela fração de encurtamento menor que 20%, aumento da separação septal do ponto E acima de 0,7cm e aumento do índice volumétrico do ventrículo esquerdo ao final da sístole (61,4ml/m²), realizaram-se os eletrocardiogramas por 24 horas. Os resultados demonstraram aumentos de 44,6% e 41,7% nas freqüências cardíacas mínima e média, respectivamente, e presença, com maior freqüência, de arritmias supraventriculares do que ventriculares nos animais do grupo B. Concluiu-se que o Holter é eficaz e demonstra, com precocidade e melhor definição, as alterações da freqüência e do ritmo cardíaco de cães com disfunção miocárdica induzida pela doxorrubicina.

The effects of anesthesia with a combination of intramuscular xylazine-diazepam-ketamine on heart rate, respiratory rate and cloacal temperature in roosters

Objective To examine the anesthetic effects of a xylazine-diazepam-ketamine (XDK) combination in roosters.Study design Prospective experimental trial.Animals Six healthy white Leghorn roosters weighing 2.03 +/- 0.08 kg.Methods Each rooster was pre-medicated with xylazine (3 mg kg(-1), IM) and after 15 minutes anesthesia was induced with a diazepam (4 mg kg(-1)) and ketamine (25 mg kg(-1)) combination injected into the pectoral muscles. Heart and respiratory rates were recorded before anesthesia and every 15 minutes after induction for 165 minutes. Cloacal temperature was measured before and 15 minutes after pre-medication and every 75 minutes thereafter during anesthesia. Quality of induction and recovery were scored subjectively; duration of loss of righting reflex, abolition of response to a painful stimulus and palpebral reflex were also recorded.Results Intramuscular injection of xylazine smoothly induced loss of the righting reflex within 3-4 minutes. Loss of response to a painful stimulus occurred at 13.1 +/- 2.9 minutes (mean +/- SD) after the administration of the D-K combination, and lasted for 63.0 +/- 5.3 minutes. Roosters anesthetized with this combination had a significant decrease in heart and respiratory rates and cloacal temperature. The recovery period lasted for up to 4 hours (227.5 +/- 15.4 minutes). Quality of recovery was satisfactory for four roosters but excitation was noted in two birds.Conclusions and clinical relevance The XDK combination was a useful anesthetic technique for typhlectomy in roosters. Nevertheless this drug combination should be used with caution and cardiopulmonary parameters monitored carefully. Under the conditions of this experiment it was associated with a decreased cloacal temperature and prolonged recoveries.

Efeito do treinamento resistido na função autônomica cardíaca, nos parâmetros bioquímicos e antropométricos de pessoas vivendo com HIV/AIDS

Introduction: The emergence of High Active Antiretroviral Therapy (HAART) increase the life expectancy of the persons living with HIV/AIDS (PLHIV), therefore the prolonged use cause metabolic implications and influences on body fat distribution and increase the cardiovascular diseases prevalence. Aims: Evaluate the effect of resistance training on heart rate variability, biochemical parameters and somatotype on PLHIV. Methods: Participated this study seven sedentary men, with age above 25 years old, living with HIV/AIDS, under HAART use. Were submitted a 16 week intervention with resistance training. Evaluated the heart rate variability, biochemical parameters and somatotype, before, after 8 weeks and 16 weeks, all in paired form. It was found the data normality by Shapiro-Wilk test and conducted the Anova one way combined with Tukey post hoc to samples in each evaluate moment, adopting significance level p<0,05. Also were calculated percentage change deltas. For somatotype was used the somatotype spatial distance (DES), obeying the significance value DES≥1. Results: Was found significance differences only in variable final heart rate delta 60s (p=0,01), however, is not showed changes on heart rate variability, biochemical parameters and somatotype components. Conclusion: 16 weeks of resistance training showed improvement on heart rate recovery after submaximal effort and, despite is not enough to produce significance differences on biochemical parameters and somatotype components, could be realize improvement on average value of fasting glucose and lipid profile, as well as reducing the endomorphic component

Higher order ionospheric effects in GNSS positioning in the European region

After removal of the Selective Availability in 2000, the ionosphere became the dominant error source for Global Navigation Satellite Systems (GNSS), especially for the high-accuracy (cm-mm) demanding applications like the Precise Point Positioning (PPP) and Real Time Kinematic (RTK) positioning.The common practice of eliminating the ionospheric error, e. g. by the ionosphere free (IF) observable, which is a linear combination of observables on two frequencies such as GPS L1 and L2, accounts for about 99% of the total ionospheric effect, known as the first order ionospheric effect (Ion1). The remaining 1% residual range errors (RREs) in the IF observable are due to the higher - second and third, order ionospheric effects, Ion2 and Ion3, respectively. Both terms are related with the electron content along the signal path; moreover Ion2 term is associated with the influence of the geomagnetic field on the ionospheric refractive index and Ion3 with the ray bending effect of the ionosphere, which can cause significant deviation in the ray trajectory (due to strong electron density gradients in the ionosphere) such that the error contribution of Ion3 can exceed that of Ion2 (Kim and Tinin, 2007).The higher order error terms do not cancel out in the (first order) ionospherically corrected observable and as such, when not accounted for, they can degrade the accuracy of GNSS positioning, depending on the level of the solar activity and geomagnetic and ionospheric conditions (Hoque and Jakowski, 2007). Simulation results from early 1990s show that Ion2 and Ion3 would contribute to the ionospheric error budget by less than 1% of the Ion1 term at GPS frequencies (Datta-Barua et al., 2008). Although the IF observable may provide sufficient accuracy for most GNSS applications, Ion2 and Ion3 need to be considered for higher accuracy demanding applications especially at times of higher solar activity.This paper investigates the higher order ionospheric effects (Ion2 and Ion3, however excluding the ray bending effects associated with Ion3) in the European region in the GNSS positioning considering the precise point positioning (PPP) method. For this purpose observations from four European stations were considered. These observations were taken in four time intervals corresponding to various geophysical conditions: the active and quiet periods of the solar cycle, 2001 and 2006, respectively, excluding the effects of disturbances in the geomagnetic field (i.e. geomagnetic storms), as well as the years of 2001 and 2003, this time including the impact of geomagnetic disturbances. The program RINEX_HO (Marques et al., 2011) was used to calculate the magnitudes of Ion2 and Ion3 on the range measurements as well as the total electron content (TEC) observed on each receiver-satellite link. The program also corrects the GPS observation files for Ion2 and Ion3; thereafter it is possible to perform PPP with both the original and corrected GPS observation files to analyze the impact of the higher order ionospheric error terms excluding the ray bending effect which may become significant especially at low elevation angles (Ioannides and Strangeways, 2002) on the estimated station coordinates.

Índices geométricos de variabilidade da frequência cardíaca na doença pulmonar obstrutiva crônica

Background: It was already evidenced decreased heart rate variability (HRV) in chronic obstructive pulmonary disease (COPD) patients at rest.Objective: In order to insert new elements in the literature regarding this issue, we evaluated geometric index of HRV in COPD subjects.Method: We analyzed data from 34 volunteers, divided into two groups according to spirometric values: COPD (17 volunteers, FEV1/FVC = 47.3 +/- 10.2; FEV1 = 50.8 +/- 15.7) and control (17 volunteers, FEV1/FVC = 78.8 +/- 10.8; FEV1 = 100.1 +/- 14.7). For analysis of HRV indexes the volunteers remained in the supine position for 30 minutes. We analyzed the following indexes: triangular index (RRtri), triangular interpolation of RR intervals (TINN) and Poincare plot (SD1, SD2 and SD1/SD2). Student t test for unpaired samples and Mann-Whitney test were used for data analysis.Results: We observed statistically significant reductions in geometric indexes in the COPD group: RRtri (0.043 +/- 0.01 vs. 0.059 +/- 0.02; p = 0.018), TINN (105.88 +/- 51.82 vs. 151.47 +/- 49.9; p=0.014), SD1 (9.76 +/- 4.66 vs. 14.55 +/- 6.04; p = 0.014) and SD2 (34.86 +/- 17.02 vs. 51.51 +/- 18.38; p = 0.010). SD1/5D2 (0.30 +/- 0.11 vs. 0.28 +/- 0.07; p = 0.605) were not significantly different between groups. Patients with COPD presented a visual analysis of Poincare plot of lower dispersion of RR intervals both beat to beat and the long term.Conclusion: Subjects with COPD present reduction of geometric indexes of HRV, indicating reduced heart rate variability. (C) 2010 Sociedade Portuguesa de Pneumologia. Published by Elsevier Espana, S.L. All rights reserved.