A Critical Analysis of the Available In Vitro

Angiogenesis is a biological process with a central role in retinal diseases. The choice of the ideal method to study angiogenesis, particularly in the retina, remains a problem. Angiogenesis can be assessed through in vitro and in vivo studies. In spite of inherent limitations, in vitro studies are faster, easier to perform and quantify, and typically less expensive and allow the study of isolated angiogenesis steps. We performed a systematic review of PubMed searching for original articles that applied in vitro or ex vivo angiogenic retinal assays until May 2017, presenting the available assays and discussing their applicability, advantages, and disadvantages. Most of the studies evaluated migration, proliferation, and tube formation of endothelial cells in response to inhibitory or stimulatory compounds. Other aspects of angiogenesis were studied by assessing cell permeability, adhesion, or apoptosis, as well as by implementing organotypic models of the retina. Emphasis is placed on how the methods are applied and how they can contribute to retinal angiogenesis comprehension. We also discuss how to choose the best cell culture to implement these methods. When applied together, in vitro and ex vivo studies constitute a powerful tool to improve retinal angiogenesis knowledge. This review provides support for researchers to better select the most suitable protocols in this field

Physical properties of single-crystalline fibers of the colossal-magnetoresistance manganite La0.7Ca0.3MnO3

We have grown high-quality single crystals of the colossal-magnetoresistance (CMR) material La0.7Ca0.3MnO3 by using the laser heated pedestal growth (LHPG) method. Samples were grown as fibers of different diameters, and with lengths of the order of centimeters. Their composition and structure were verified through X-ray diffraction, scanning electron microcopy with EDX (Energy Dispersive X-ray Analysis) and by Rietveld analysis. The quality of the crystalline fibers was confirmed by Laue and EBSD (Electron Backscatter Diffraction) patterns. Rocking curves performed along the fiber axis revealed a half-height width of 0.073 degrees. The CMR behavior was confirmed by electrical resistivity and magnetization measurements as a function of temperature.Comment: 11 pages (including 3 figures); to appear in Appl. Phys. Let

Moderate exercise training provides left ventricular tolerance to acute pressure overload

Moreira-Goncalves D, Henriques-Coelho T, Fonseca H, Ferreira RM, Amado F, Leite-Moreira A, Duarte JA. Moderate exercise training provides left ventricular tolerance to acute pressure overload. Am J Physiol Heart Circ Physiol 300: H1044-H1052, 2011. First published December 24, 2010; doi: 10.1152/ajpheart.01008.2010.-The present study evaluated the impact of moderate exercise training on the cardiac tolerance to acute pressure overload. Male Wistar rats were randomly submitted to exercise training or sedentary lifestyle for 14 wk. At the end of this period, the animals were anaesthetized, mechanically ventilated, and submitted to hemodynamic evaluation with biventricular tip pressure manometers. Acute pressure overload was induced by banding the descending aorta to induce a 60% increase of peak systolic left ventricular pressure during 120 min. This resulted in the following experimental groups: 1) sedentary without banding (SED + Sham), 2) sedentary with banding (SED + Band), and 3) exercise trained with banding (EX + Band). In response to aortic banding, SED + Band animals could not sustain the 60% increase of peak systolic pressure for 120 min, even with additional narrowing of the banding. This was accompanied by a reduction of dP/dt(max) and dP/dt(min) and a prolongation of the time constant tau, indicating impaired systolic and diastolic function. This impairment was not observed in EX + Band (P < 0.05 vs. SED + Band). Additionally, compared with SED + Band, EX + Band presented less myocardial damage, exhibited attenuated protein expression of active caspase-3 and NF-kappa B (P < 0.016), and showed less protein carbonylation and nitration (P < 0.05). These findings support our hypothesis that exercise training has a protective role in the modulation of the early cardiac response to pressure overload

Identification of suitable adjuvant for vaccine formulation with the Neospora caninum antigen NcSRS2.

The parasite Neospora caninum is the main cause of abortion in cattle in many countries around the world, so a vaccine is a rational approach method for the control of the disease. An effective vaccine should be able to prevent both, the horizontal and vertical transmission of N. caninum. In this study, the immune vaccinal response of the recombinant protein rNcSRS2 of N. caninum expressed in Pichia pastoris and formulated with water-in-oil emulsion, xanthan gum, and alum hydroxide was assessed in an experimental murine model. Groups of 10 Balb/c mice were subcutaneously inoculated with two doses of prNcSRS2 twenty-one days apart. After the second immunization, four mice from each group were euthanized, and splenocytes were stimulated ex vivo with recombinant protein. The IgG dynamics were evaluated by indirect ELISA, and the splenocytes cytokines transcription by qPCR. All groups elicited specific antibodies against prNcSRS2, with the water-in-oil group showing significantly (p .05) elevated titers compared to the other groups. The prNcSRS2 protein alone did not induce a significant ex vivo splenic transcription level of IFN-c, TNF-a, IL-4, IL-10, and IL-12 cytokines, except for IL-17A, and the adjuvant associations with the prNcSRS2 protein induced different cytokine transcription profiles. The water-inoil emulsion modulated the expression of TNF-a; the xanthan gum modulated IL-4, IL-10, and IL-12; and alum hydroxide modulated IFN-c, TNF-a, IL-4, IL-10, and IL-12. In conclusion, it was found that the association of the recombinant prNcSRS2 protein with different adjuvants induced different levels of specific antibody, and a distinct splenic cytokine profile in an adjuvant-dependent manner. The mechanisms of adjuvancity activity is complex, so adjuvant formulation may help in the design of efficient vaccine to control Neosporosis

Monitoramento e determinação do nível de ação do ácaro-rajado na cultura da uva.

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Frailty-Independent Undertreatment Negative Impact on Survival in Older Patients With Breast Cancer

Purpose: The management of older adults with breast cancer (BC) remains controversial. The challenging assessment of aging idiosyncrasies and the scarce evidence of therapeutic guidelines can lead to undertreatment. Our goal was to measure undertreatment and assess its impact on survival. Methods: Consecutive patients with BC aged 70 years or older were prospectively enrolled in 2014. Three frailty screening tools (G8, fTRST, and GFI) and two functional status scales (Karnofsky performance score and Eastern Cooperative Oncology Group Performance Status) were applied. Disease characteristics, treatment options, and causes of mortality were recorded during a 5-year follow-up. In addition, we defined undertreatment and correlated its survival impact with frailty. Results: A total of 92 patients were included in the study. The median age was 77 (range 70-94) years. The prevalence of frailty was discordant (G8, 41.9%; fTRST, 74.2%; GFI, 32.3%). Only 47.8% of the patients had a local disease, probably due to a late diagnosis (73.9% based on self-examination). Thirty-three patients (35.6%) died, of which 15 were from BC. We found a considerably high proportion (53.3%) of undertreatment, which had a frailty-independent negative impact on the 5-year survival (hazard ratio [HR], 5.1; 95% confidence interval [CI], 2.1-12.5). Additionally, omission of surgery had a frailty-independent negative impact on overall survival (HR, 3.9; 95% CI, 1.9-7.9). Conclusion: BC treatment in older adults should be individualized. More importantly, assessing frailty (not to treat) is essential to be aware of the risk-benefit profile and the patient's well-informed willingness to be treated. Undertreatment in daily practice is frequent and might have a negative impact on survival, as we report.This article publication was supported by National Funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within CINTESIS, R&D Unit (reference UIDB/4255/2020)

Impact of the 2016 ASE/EACVI recommendations on the prevalence of diastolic dysfunction in the general population

Aims: Diastolic dysfunction (DD) is frequent in the general population; however, the assessment of diastolic function remains challenging. We aimed to evaluate the impact of the recent 2016 American Society of Echocardiography (ASE)/European Association of Cardiovascular Imaging (EACVI) recommendations in the prevalence and grades of DD compared with the 2009 guidelines and the Canberra Study Criteria (CSC). Methods and results: Within a population-based cohort, a total of 1000 individuals, aged ≥45 years, were evaluated retrospectively. Patients with previously known cardiac disease or ejection fraction <50% were excluded. Diastolic function was assessed by transthoracic echocardiography. DD prevalence and grades were determined according to the three classifications. The mean age was 62.0 ± 10.5 years and 37% were men. The prevalence of DD was 1.4% (n = 14) with the 2016 recommendations, 38.1% (n = 381) with the 2009 recommendations, and 30.4% (n = 304) using the CSC. The concordance between the updated recommendations and the other two was poor (from k = 0.13 to k = 0.18, P < 0.001). Regarding the categorization in DD grades, none of the 14 individuals with DD by the 2016 guidelines were assigned to Grade 1 DD, 64% were classified as Grade 2, 7% had Grade 3, and 29% had indeterminate grade. Conclusion: The application of the new 2016 ASE/EACVI recommendations resulted in a much lower prevalence of DD. The concordance between the classifications was poor. The updated algorithm seems to be able to diagnose only the most advanced cases

Left atrial deformation analysis by speckle tracking echocardiography to predict exercise capacity after myocardial infarction

Introduction: Left atrial (LA) size and function are associated with outcome after myocardial infarction (MI). In this study we aimed to assess the impact of LA function as a predictor of exercise capacity through speckle tracking echocardiography. Methods: A total of 94 patients (mean age 54.8 +/- 11.0 years; 82% male) were enrolled one month after MI. Echocardiography was used to assess LA volumes and various indices of LA conduit, contraction and reservoir function. LA deformation was assessed by two-dimensional speckle tracking to calculate strain and strain rate at different phases of the cardiac cycle. Exercise capacity was assessed by oxygen uptake (VO2) on cardiopulmonary exercise testing. Results: Increased LA volumes, especially LA volume before atrial contraction, were correlated with reduced peak VO2 and reduced VO2 at anaerobic threshold. Decreased peak VO2 was associated with reduced LA conduit function (p=0.24; p=0.02), but not with LA booster function (p=-0.07; p=0.53). Lower peak atrial longitudinal strain was associated with worse exercise capacity (p=0.24; p=0.02). Conclusions: After MI, increased LA volumes were markers of decreased functional capacity that was associated with decreased LA conduit function, but not with LA contractile function. In these patients, LA longitudinal strain analysis may be useful to predict reduced exercise capacity. (C) 2018 Sociedade Portuguesa de Cardiologia. Published by Elsevier Espana, S.L.U.This work was supported by the Portuguese Foundation for Science and Technology grants PEst-C/SAU/UI0051/2011 and EXCL/BIM-MEC/0055/2012 through the Cardiovascular R&D Unit and by European Commission grant FP7-Health-2010; MEDIA-261409