Role of MEF2C in the Endothelial Cells Derived from Human Induced Pluripotent Stem Cells.

Human induced pluripotent stem cells (hiPSCs) not only provide an abundant source of vascular cells for potential therapeutic applications in vascular disease but also constitute an excellent model for understanding the mechanisms that regulate the differentiation and the functionality of vascular cells. Here, we reported that myocyte enhancer factor 2C (MEF2C) transcription factor, but not any other members of the MEF2 family, was robustly upregulated during the differentiation of vascular progenitors and endothelial cells (ECs) from hiPSCs. Vascular endothelial growth factors (VEGF) strongly induced MEF2C expression in endothelial lineage cells. The specific upregulation of MEF2C during the commitment of endothelial lineage was dependent on the extracellular signal regulated kinase (ERK). Moreover, knockdown of MEF2C with shRNA in hiPSCs did not affect the differentiation of ECs from these hiPSCs, but greatly reduced the migration and tube formation capacity of the hiPSC-derived ECs. Through a chromatin immunoprecipitation-sequencing, genome-wide RNA-sequencing, quantitative RT-PCR, and immunostaining analyses of the hiPSC-derived endothelial lineage cells with MEF2C inhibition or knockdown compared to control hiPSC-derived ECs, we identified TNF-related apoptosis inducing ligand (TRAIL) and transmembrane protein 100 (TMEM100) as novel targets of MEF2C. This study demonstrates an important role for MEF2C in regulating human EC functions and highlights MEF2C and its downstream effectors as potential targets to treat vascular malfunction-associated diseases.

Molecular survey of Zika virus in the animal-human interface in traditional farming.

Backyard animal husbandry is common in rural communities in developing countries and, given the conditions in which it occurs, it can increase the risk of disease transmission, such as arboviruses. To determine the presence of the Zika virus (ZIKV) and abundance of its arthropod vectors we evaluated the socioeconomic implications involved in its transmission in two highly vulnerable Mayan communities in the state of Yucatan that practice backyard farming. An analytical cross-sectional study was carried out throughout 2016 to understand socioeconomic variables and seasonal patterns in mosquito populations. We selected 20 households from each community. Social exclusion indicators were analyzed, human and domestic animals were sampled, and mosquitoes were collected and identified. Four out of eight indicators of social exclusion were higher than the reported national averages. We captured 5,825 mosquitoes from 16 species being Culex quinquefasciatus and Aedes aegypti the most abundant. The presence of chickens and human overcrowding in dwellings were the most significant factors (P = 0.026) associated with the presence of Ae. aegypti. Septic tanks (odds ratio = 6.64) and chickens (odds ratio = 27.41) in backyards were the main risk factors associated with the presence of immature states of Ae. aegypti in both communities. Molecular analysis to detect ZIKV was performed in blood samples from 416 humans, 1,068 backyard animals and 381 mosquito pools. Eighteen humans and 10 pig pools tested positive for ZIKV. Forty-three mosquito pools tested positive for flavivirus. Ten of the 43 pools of positive mosquitoes were sequenced, corresponding 3/10 to ZIKV and 1/10 to Dengue virus type 2. The findings obtained indicate the continuous circulation of Flavivirus (including ZIKV) in backyard environments in vulnerable communities, highlighting the importance of studying their transmission and maintenance in these systems, due that backyard animal husbandry is a common practice in these vulnerable communities with limited access to health services.

Electroanalytical Enzyme Biosensor Based on Cordia superba Enzyme Extract for the Detection of Phytomarkers in Kombucha.

Antioxidants are responsible for many beneficial health effects and are highly present in natural products, such as kombucha. Biosensors' development targeting antioxidants and phytomarkers are an active research field. This work aimed to propose a voltammetric polyphenolxidase (Cordia superba) biosensor for catechin and total phenolic compounds quantification in kombucha samples. Optimizations were performed on the biosensor of Cordia superba to improve the accuracy and selectivity, such as enzyme-substrate interaction time, analytical responses for different patterns and signal differences with the carbon paste and modified carbon paste electrode. Kombucha probiotic drink samples were fermented for 7 to 14 days at a controlled temperature (28 ± 2 °C). A linear curve was made for catechin with a range of 10.00 to 60.00 µM, with a limit of detection of 0.13 µM and limit of quantification of 0.39 µM. The biosensor proposed in this work was efficient in determining the patterns of phenolic compounds in kombucha.

Evaluation of Antioxidant Potential of Commercial Cinnamon Samples and Its Vasculature Effects.

Growing concerns on free radicals are the oxidative processes associated with physiological damage. The consumption of functional foods and use of plants with antioxidant capacity are widespread. Given the importance of determining antioxidant capacity in relation to the therapeutic effect, this study was aimed at evaluating cinnamon extract (Cinnamomum sp.) in commercial samples by spectrophotometric and voltammetric methods and assessing the vascular activity of some samples. The spectrophotometric methods performed were DPPH (1,1-diphenyl-2-picrihydrazine), ABTS (2,21-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)), and Folin-Ciocalteu radical sequestration assays. For the electrochemical experiments, a three-electrode system was used, consisting of carbon paste electrode, platinum wire, and Ag/AgCl/KClsat, representing the working, auxiliary, and reference electrodes, respectively. The electroanalytical methods used were differential pulse, square wave, and cyclic voltammetries. The extracts were prepared in hydroalcoholic solution. A calibration curve with gallic acid was calculated to quantify their equivalent amounts in the analyzed extract. The correlation between the electrochemical approach and the total phenols calculated by the ABTS, DPPH, and Folin-Ciocalteu methods was 0.63, 0.7, and 0.73, respectively, with 1 being an ideal directly proportional correlation. The correlation between spectrophotometric methods was 0.83. A biosensor was developed in a carbon paste electrode using the enzyme laccase, obtained by the fungus Marasmiellus colocasiae. It was observed that the antioxidant profile of the cinnamon samples had an analytical sign improvement of up to 4 times when compared with the electrode without the modification. The samples were analyzed by mass spectrometer, and the main chemical markers found were coumarin, cinnamaldehyde, and eugenol. Pharmacological trials showed that these samples also induce a significant vasorelaxant effect associated to antioxidant potential on vascular injury induced by oxidative stress. Thus, cinnamon showed a high antioxidant capacity, in agreement with the results obtained in other studies, emphasizing its importance as a functional food.

Enzymatic Electroanalytical Biosensor Based on Maramiellus colocasiae Fungus for Detection of Phytomarkers in Infusions and Green Tea Kombucha.

In this work, we developed an enzymatic voltammetric biosensor for the determination of catechin and gallic acid in green tea and kombucha samples. The differential pulse voltammetry (DPV) methodology was optimized regarding the amount of crude enzyme extract, incubation time in the presence of the substrates, optimal pH, reuse of the biosensor, and storage time. Samples of green tea and kombucha were purchased in local markets in the city of Goiânia-GO, Brazil. High performance liquid chromatography (HPLC) and Folin-Ciocalteu spectrophotometric techniques were performed for the comparison of the analytical methods employed. In addition, two calibration curves were made, one for catechin with a linear range from 1 to 60 µM (I = -0.152 * (catechin) - 1.846), with a detection limit of 0.12 µM and a quantification limit of 0.38 µM and one for gallic acid with a linear range from 3 to 60 µM (I = -0.0415 * (gallic acid) - 0.0572), with a detection limit of 0.14 µM and a quantification limit of 0.42 µM. The proposed biosensor was efficient in the determination of phenolic compounds in green tea.

Evidence of Zika Virus Infection in Pigs and Mosquitoes, Mexico.

Evidence suggests that pigs seroconvert after experimental exposure to Zika virus and are potential sentinels. We demonstrate that pigs are also susceptible to natural Zika virus infection, shown by the presence of antibodies in domestic pigs in Yucatan, Mexico. Zika virus RNA was detected in 5 species of mosquitoes collected inside pigpens.

Mechanoregulation of p38 activity enhances endoplasmic reticulum stress-mediated inflammation by arterial endothelium.

Endothelial up-regulation of VCAM-1 at susceptible sites in arteries modulates the recruitment efficiency of inflammatory monocytes that initiates atherosclerotic lesion formation. We reported that hydrodynamic shear stress (SS) mechanoregulates inflammation in human aortic endothelial cells through endoplasmic reticulum (ER) stress via activation of the transcription factor x-box binding protein 1 (XBP1). Here, a microfluidic flow channel that produces a linear gradient of SS along a continuous monolayer of endothelium was used to delve the mechanisms underlying transcriptional regulation of TNF-α-stimulated VCAM-1 expression. High-resolution immunofluorescence imaging enabled continuous detection of platelet endothelial cell adhesion molecule 1 (PECAM-1)-dependent, outside-in signaling as a function of SS magnitude. Differential expression of VCAM-1 and intercellular adhesion molecule 1 (ICAM-1) was regulated by the spatiotemporal activation of MAPKs, ER stress markers, and transcription factors, which was dependent on the mechanosensing of SS through PECAM-1 and PI3K. Inhibition of p38 specifically abrogated the rise to peak VCAM-1 at low SS (2 dyn/cm2), whereas inhibition of ERK1/2 attenuated peak ICAM-1 at high SS (12 dyn/cm2). A shear stress-regulated temporal rise in p38 phosphorylation activated the nuclear translocation of XBP1, which together with the transcription factor IFN regulatory factor 1, promoted maximum VCAM-1 expression. These data reveal a mechanism by which SS sensitizes the endothelium to a cytokine-induced ER stress response to spatially regulate inflammation promoting atherosclerosis.-Bailey, K. A., Moreno, E., Haj, F. G., Simon, S. I., Passerini, A. G. Mechanoregulation of p38 activity enhances endoplasmic reticulum stress-mediated inflammation by arterial endothelium.

Voltammetric Evaluation of Diclofenac Tablets Samples through Carbon Black-Based Electrodes.

Diclofenac (DIC) is a non-steroidal anti-inflammatory drug of wide use around the world. Electroanalytical methods display a high analytical potential for application in pharmaceutical samples but the drawbacks concerning electrode fouling and reproducibility are of major concern. Henceforth, the aim of this work was to propose the use of alternative low-cost carbon black (CB) and ionic liquid (IL) matrix to modify the surface of pencil graphite electrodes (PGE) in order to quantify DIC in raw materials, intermediates, and final products, as well as in stability assays of tablets. The proposed method using CB+IL/PGE displayed good recovery (99.4%) as well as limits of detection (LOD) of 0.08 µmol L-1 and limits of quantification (LOQ) of 0.28 µmol L-1. CB+IL/PGE response was five times greater than the unmodified PGE. CB+IL-PGE stands as an interesting alternative for DIC assessment in different pharmaceutical samples.

Ecotoxicological assessment and electrochemical remediation of doxorubicin.

Doxorubicin (DOX) is an anthracycline widely used in treatments of several cancers, so it has found in hospital effluents with a significant concentration (above 1 µg L-1). Electrochemical remediation is an alternative to promote its degradation. The aim of this work was to evaluate the ability of nanostructured graphite electrodes with metallic oxides to degrade DOX by electro-oxidation (EO). Graphite, TiO2@graphite and AuO-TiO2@graphite electrodes were used in medium with tap water or 10 mmol L-1 NaCl. DOX treatments at concentrations of 1.25-5 mg L-1 were carried out in a voltage source with 1.5-5 V. The cathode used was the platinum electrode. The treatment of DOX 1.25 mg L-1 with 10 mmol L-1 NaCl electrolyte using the AuO-TiO2@graphite electrode at 5 V and 1 mA was the best methodology to promote its degradation. Also, the modified electrode was efficient to DOX degradation after 17 cycles of reuse. An energy expenditure of 1.11 and 0.2 kWh m-3 were obtained for 3 and 50 mL of treatment, respectively. Fish embryo acute toxicity test with zebrafish (Danio rerio) were performed before and after treatment by EO using NaCl. This treatment caused no effect on embryo-larval development, however it induced significant damage in the DNA of the zebrafish larvae after 96 h of exposure, which emphasizes the importance of a depth ecotoxicological evaluation during the development of EO methodologies.

Nanostructured TiO2 Carbon Paste Based Sensor for Determination of Methyldopa.

Methyldopa is a catecholamine widely used in the treatment of mild to moderate hypertension whose determination in pharmaceutical formulae is of upmost importance for dose precision. Henceforth, a low-cost carbon paste electrode (CPE) consisting of graphite powder obtained from a crushed pencil stick was herein modified with nanostructured TiO2 (TiO2@CPE) aiming for the detection of methyldopa in pharmaceutical samples. The TiO2-modified graphite powder was characterized by scanning electron microscopy and X-ray diffraction, which demonstrated the oxide nanostructured morphology. Results evidenced that sensitivity was nonetheless increased due to electro-catalytic effects promoted by metal modification, and linear response obtained by differential pulse voltammetry for the determination of methyldopa (pH = 5.0) was between 10⁻180 µmol/L (Limit of Detection = 1 µmol/L) with the TiO2@CPE sensor. Furthermore, the constructed sensor was successfully applied in the detection of methyldopa in pharmaceutical formulations and excipients promoted no interference, that indicates that the sensor herein developed is a cheap, reliable, and useful strategy to detect methyldopa in pharmaceutical samples, and may also be applicable in determinations of similar compounds.

Antioxidant activity evaluation of dried herbal extracts: an electroanalytical approach

ABSTRACT The prevention of chronic and degenerative diseases, is a health concern deeply associated with oxidative stress. Such progressive phenomena can be avoided through exogenous antioxidant intake, which set up a reductant cascade, mopping up damaging free radicals. Medicinal herbs are commonly associated with high antioxidant potential, and hence their health benefits. The commerce of dried herbal extracts movements a big portion of developing countries economy. The determination of medicinal herbs the antioxidant activity capacity is of utmost importance. The assessment of antioxidant activity in phytotherapics is mostly achieved by spectrophotometric assays, however colored substances can produce interferences that do not occur in electroanalytical methods. Therefore, the aim of this paper is to compare spectrophotometric and voltammetric techniques to evaluate antioxidant activity in herbal drugs such as: Ginkgo biloba L., Camellia sinensis (L.) Kuntze, Theaceae; Hypericum perforatum L., Hypericaceae; Aesculus hippocastanum L., Sapindaceae; Rosmarinus officinalis L., Lamiaceae; Morinda citrifolia L., Rubiaceae; Centella asiatica (L.) Urb., Apiaceae; Trifolium pratense L., Fabaceae; Crataegus oxyacantha L., Rosaceae; and Vaccinium macrocarpon Aiton, Ericaceae. The spectrophotometric methods employed were DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and the Folin-Ciocalteu assays. The electroanalytical method used was voltammetry and it was developed a phenoloxidase based biosensor. The redox behavior observed for each herbal sample resulted in distinguishable voltammetric profiles. The highest electrochemical indexes were found to G. biloba and H. perforatum, corroborating to traditional spectrophotometric methods. Thus, the electroanalysis of herbal drugs, may be a promising tool for antioxidant potential assessment.

"Give me some space": exploring youth to parent aggression and violence.

A small scale qualitative project, undertaken by an interdisciplinary domestic violence research group involving academic researchers and research assistants, with colleagues from Independent Domestic Abuse Services (IDAS), investigated youth aggression and violence against parents. Following the literature review, data was generated through several research conversations with young people (n = 2), through semi-structured interviews with mothers (n = 3) and practitioners (n = 5), and through a practitioner focus group (n = 8). Thematic analysis and triangulation of the data from parents, practitioners and young people, elicited interconnected and complex overarching themes. Young people could be both victim and perpetrator. The witnessing or experiencing of domestic aggression and violence raised the concept of 'bystander children'. The impact of young people experiencing familial violence was underestimated by parents. For practitioners, the effects of working with domestic violence was shown to be significant - both positively and negatively.

Loss of mouse cardiomyocyte talin-1 and talin-2 leads to ß-1 integrin reduction, costameric instability, and dilated cardiomyopathy.

Continuous contraction-relaxation cycles of the heart require strong and stable connections of cardiac myocytes (CMs) with the extracellular matrix (ECM) to preserve sarcolemmal integrity. CM attachment to the ECM is mediated by integrin complexes localized at the muscle adhesion sites termed costameres. The ubiquitously expressed cytoskeletal protein talin (Tln) is a component of muscle costameres that links integrins ultimately to the sarcomere. There are two talin genes, Tln1 and Tln2. Here, we tested the function of these two Tln forms in myocardium where Tln2 is the dominant isoform in postnatal CMs. Surprisingly, global deletion of Tln2 in mice caused no structural or functional changes in heart, presumably because CM Tln1 became up-regulated. Tln2 loss increased integrin activation, although levels of the muscle-specific ß1D-integrin isoform were reduced by 50%. With this result, we produced mice that had simultaneous loss of both CM Tln1 and Tln2 and found that cardiac dysfunction occurred by 4 wk with 100% mortality by 6 mo. ß1D integrin and other costameric proteins were lost from the CMs, and membrane integrity was compromised. Given that integrin protein reduction occurred with Tln loss, rescue of the phenotype was attempted through transgenic integrin overexpression, but this could not restore WT CM integrin levels nor improve heart function. Our results show that CM Tln2 is essential for proper ß1D-integrin expression and that Tln1 can substitute for Tln2 in preserving heart function, but that loss of all Tln forms from the heart-muscle cell leads to myocyte instability and a dilated cardiomyopathy.

Concentrations of Polychlorinated Biphenyls and Organochlorine Pesticides in Umbilical Cord Blood Serum of Newborns in Kingston, Jamaica.

To date much of the biomonitoring related to exposure to polychlorinated biphenyls (PCBs) and organochlorine (OC) pesticides is from middle to high income countries, including the U.S., Canada and Europe, but such data are lacking for the majority of low to middle income countries. Using data from 64 pregnant mothers who were enrolled in 2011, we aimed to assess the concentrations of the aforementioned toxins in umbilical cord blood serum of 67 Jamaican newborns. For 97 of the 100 PCB congeners and 16 of the 17 OC pesticides, all (100%) concentrations were below their respective limits of detection (LOD). Mean (standard deviation (SD)) lipid-adjusted concentrations in cord blood serum for congeners PCB-153, PCB-180, PCB-206 and total PCB were 14.25 (3.21), 7.16 (1.71), 7.30 (1.74) and 28.15 (6.03) ng/g-lipid, respectively. The means (SD) for the 4,4'-dichlorodiphenyldichloroethylene (DDE)-hexane fraction and total-DDE were 61.61 (70.78) and 61.60 (70.76) ng/g-lipid, respectively. Compared to the U.S. and Canada, the concentrations of these toxins were lower in cord-blood serum of Jamaican newborns. We discuss that these differences could be partly due to differences in dietary patterns in these countries. Despite limitations in our dataset, our results provide information on the investigated toxins in cord blood serum that could serve as a reference for Jamaican newborns.