Amyloid ß-Oligomers Inhibit the Nuclear Ca2+ Signals and the Neuroprotective Gene Expression Induced by Gabazine in Hippocampal Neurons.

Hippocampal neuronal activity generates dendritic and somatic Ca2+ signals, which, depending on stimulus intensity, rapidly propagate to the nucleus and induce the expression of transcription factors and genes with crucial roles in cognitive functions. Soluble amyloid-beta oligomers (AßOs), the main synaptotoxins engaged in the pathogenesis of Alzheimer's disease, generate aberrant Ca2+ signals in primary hippocampal neurons, increase their oxidative tone and disrupt structural plasticity. Here, we explored the effects of sub-lethal AßOs concentrations on activity-generated nuclear Ca2+ signals and on the Ca2+-dependent expression of neuroprotective genes. To induce neuronal activity, neuron-enriched primary hippocampal cultures were treated with the GABAA receptor blocker gabazine (GBZ), and nuclear Ca2+ signals were measured in AßOs-treated or control neurons transfected with a genetically encoded nuclear Ca2+ sensor. Incubation (6 h) with AßOs significantly reduced the nuclear Ca2+ signals and the enhanced phosphorylation of cyclic AMP response element-binding protein (CREB) induced by GBZ. Likewise, incubation (6 h) with AßOs significantly reduced the GBZ-induced increases in the mRNA levels of neuronal Per-Arnt-Sim domain protein 4 (Npas4), brain-derived neurotrophic factor (BDNF), ryanodine receptor type-2 (RyR2), and the antioxidant enzyme NADPH-quinone oxidoreductase (Nqo1). Based on these findings we propose that AßOs, by inhibiting the generation of activity-induced nuclear Ca2+ signals, disrupt key neuroprotective gene expression pathways required for hippocampal-dependent learning and memory processes.

Cancer History Avoids the Increase of Senescence Markers in Peripheral Cells of Amnestic Mild Cognitive Impaired Patients.

Epidemiological studies show that having a history of cancer protects from the development of Alzheimer's Disease (AD), and vice versa, AD protects from cancer. The mechanism of this mutual protection is unknown. We have reported that the peripheral blood mononuclear cells (PBMC) of amnestic cognitive impairment (aMCI) and Alzheimer's Disease (AD) patients have increased susceptibility to oxidative cell death compared to control subjects, and from the opposite standpoint a cancer history is associated with increased resistance to oxidative stress cell death in PBMCs, even in those subjects who have cancer history and aMCI (Ca + aMCI). Cellular senescence is a regulator of susceptibility to cell death and has been related to the pathophysiology of AD and cancer. Recently, we showed that cellular senescence markers can be tracked in PBMCs of aMCI patients, so we here investigated whether these senescence markers are dependent on having a history of cancer. Senescence-associated ßeta-galactosidase (SA-ß-Gal) activity, G0-G1 phase cell-cycle arrest, p16 and p53 were analyzed by flow cytometry; phosphorylated H2A histone family member X (γH2AX) by immunofluorescence; IL-6 and IL-8 mRNA by qPCR; and plasmatic levels by ELISA. Senescence markers that were elevated in PBMCs of aMCI patients, such as SA-ß-Gal, Go-G1 arrested cells, IL-6 and IL-8 mRNA expression, and IL-8 plasmatic levels, were decreased in PBMCs of Ca + aMCI patients to levels similar to those of controls or of cancer survivors without cognitive impairment, suggesting that cancer in the past leaves a fingerprint that can be peripherally traceable in PBMC samples. These results support the hypothesis that the senescence process might be involved in the inverse association between cancer and AD.

Senescence Markers in Peripheral Blood Mononuclear Cells in Amnestic Mild Cognitive Impairment and Alzheimer's Disease.

Recent studies suggest that cellular senescence plays a role in Alzheimer's Disease (AD) pathogenesis. We hypothesize that cellular senescence markers might be tracked in the peripheral tissues of AD patients. Senescence hallmarks, including altered metabolism, cell-cycle arrest, DNA damage response (DDR) and senescence secretory associated phenotype (SASP), were measured in peripheral blood mononuclear cells (PBMCs) of healthy controls (HC), amnestic mild cognitive impairment (aMCI) and AD patients. Senescence-associated ßeta-galactosidase (SA-ß-Gal) activity, G0-G1 phase cell-cycle arrest, p16 and p53 were analyzed by flow cytometry, while IL-6 and IL-8 mRNA were analyzed by qPCR, and phosphorylated H2A histone family member X (γH2AX) was analyzed by immunofluorescence. Senescent cells in the brain tissue were determined with lipofuscin staining. An increase in the number of senescent cells was observed in the frontal cortex and hippocampus of advanced AD patients. PBMCs of aMCI patients, but not in AD, showed increased SA-ß-Gal compared with HCs. aMCI PBMCs also had increased IL-6 and IL8 mRNA expression and number of cells arrested at G0-G1, which were absent in AD. Instead, AD PBMCs had significantly increased p16 and p53 expression and decreased γH2Ax activity compared with HC. This study reports that several markers of cellular senescence can be measured in PBMCs of aMCI and AD patients.

Analyzing Olfactory Neuron Precursors Non-Invasively Isolated through NADH FLIM as a Potential Tool to Study Oxidative Stress in Alzheimer's Disease.

Among all the proposed pathogenic mechanisms to understand the etiology of Alzheimer's disease (AD), increased oxidative stress seems to be a robust and early disease feature where many of those hypotheses converge. However, despite the significant lines of evidence accumulated, an effective diagnosis and treatment of AD are not yet available. This limitation might be partially explained by the use of cellular and animal models that recapitulate partial aspects of the disease and do not account for the particular biology of patients. As such, cultures of patient-derived cells of peripheral origin may provide a convenient solution for this problem. Peripheral cells of neuronal lineage such as olfactory neuronal precursors (ONPs) can be easily cultured through non-invasive isolation, reproducing AD-related oxidative stress. Interestingly, the autofluorescence of key metabolic cofactors such as reduced nicotinamide adenine dinucleotide (NADH) can be highly correlated with the oxidative state and antioxidant capacity of cells in a non-destructive and label-free manner. In particular, imaging NADH through fluorescence lifetime imaging microscopy (FLIM) has greatly improved the sensitivity in detecting oxidative shifts with minimal intervention to cell physiology. Here, we discuss the translational potential of analyzing patient-derived ONPs non-invasively isolated through NADH FLIM to reveal AD-related oxidative stress. We believe this approach may potentially accelerate the discovery of effective antioxidant therapies and contribute to early diagnosis and personalized monitoring of this devastating disease.

Sequential extraction of phenolics and pectin from mango peel assisted by ultrasound.

The present work evaluated the use of ultrasound to extract sequentially phenolics and pectin from mango peel. Initially, the influence of ethanol and ultrasound on the phenolics extraction was investigated. The results showed that the ultrasound did not affect the extraction yield of these compounds. The best total phenolics yield (67%) was obtained with an extraction solution consisting of 50% of ethanol in water (v/v) and without ultrasound application, according to the experimental design. As an innovative extraction methodology, the residue of this extraction was then used to extract pectin assisted by ultrasound. The use of ultrasound increased over than 50% of the pectin extraction yield and did not affect its quality, measured by the galacturonic acid content and the degree of esterification. The sequential extraction of phenolics and pectin shows to be an alternative to use the whole residue from mango peel.

The signaling pathways underlying BDNF-induced Nrf2 hippocampal nuclear translocation involve ROS, RyR-Mediated Ca2+ signals, ERK and PI3K.

The neurotrophin Brain-Derived Neurotrophic Factor (BDNF) induces complex neuronal signaling cascades that are critical for the cellular changes underlying synaptic plasticity. These pathways include activation of Ca2+ entry via N-methyl-D-aspartate receptors and sequential activation of nitric oxide synthase and NADPH oxidase, which via generation of reactive nitrogen/oxygen species stimulate Ca2+-induced Ca2+ release mediated by Ryanodine Receptor (RyR) channels. These sequential events underlie BDNF-induced spine remodeling and type-2 RyR up-regulation. In addition, BDNF induces the nuclear translocation of the transcription factor Nrf2, a master regulator of antioxidant protein expression that protects cells against the oxidative damage caused by injury and inflammation. To investigate the possible BDNF-induced signaling cascades that mediate Nrf2 nuclear translocation in primary hippocampal cultures, we tested here whether reactive oxygen species, RyR-mediated Ca2+ release, ERK or PI3K contribute to this response. We found that pre-incubation of cultures with inhibitory ryanodine to suppress RyR-mediated Ca2+ release, with the reducing agent N-acetylcysteine or with inhibitors of ERK or PI3K activity, prevented the nuclear translocation of Nrf2 induced by incubation for 6 h with BFNF. Based on these combined results, we propose that the key role played by BDNF as an inducer of neuronal antioxidant responses, characterized by BDNF-induced Nfr2 nuclear translocation, entails crosstalk between reactive oxygen species and RyR-mediated Ca2+ release, and the participation of ERK and PI3K activities.

Calcium Release Mediated by Redox-Sensitive RyR2 Channels Has a Central Role in Hippocampal Structural Plasticity and Spatial Memory.

AIMS: Previous studies indicate that hippocampal synaptic plasticity and spatial memory processes entail calcium release from intracellular stores mediated by ryanodine receptor (RyR) channels. In particular, RyR-mediated Ca2+ release is central for the dendritic spine remodeling induced by brain-derived neurotrophic factor (BDNF), a neurotrophin that stimulates complex signaling pathways leading to memory-associated protein synthesis and structural plasticity. To examine if upregulation of ryanodine receptor type-2 (RyR2) channels and the spine remodeling induced by BDNF entail reactive oxygen species (ROS) generation, and to test if RyR2 downregulation affects BDNF-induced spine remodeling and spatial memory. RESULTS: Downregulation of RyR2 expression (short hairpin RNA [shRNA]) in primary hippocampal neurons, or inhibition of nitric oxide synthase (NOS) or NADPH oxidase, prevented agonist-mediated RyR-mediated Ca2+ release, whereas BDNF promoted cytoplasmic ROS generation. RyR2 downregulation or inhibitors of N-methyl-d-aspartate (NMDA) receptors, or NOS or of NADPH oxidase type-2 (NOX2) prevented RyR2 upregulation and the spine remodeling induced by BDNF, as did incubation with the antioxidant agent N-acetyl l-cysteine. In addition, intrahippocampal injection of RyR2-directed antisense oligodeoxynucleotides, which caused significant RyR2 downregulation, caused conspicuous defects in a memorized spatial memory task. INNOVATION: The present novel results emphasize the key role of redox-sensitive Ca2+ release mediated by RyR2 channels for hippocampal structural plasticity and spatial memory. CONCLUSION: Based on these combined results, we propose (i) that BDNF-induced RyR2-mediated Ca2+ release and ROS generation via NOS/NOX2 are strictly required for the dendritic spine remodeling and the RyR2 upregulation induced by BDNF, and (ii) that RyR2 channel expression is crucial for spatial memory processes. Antioxid. Redox Signal. 29, 1125-1146.

RyR2-Mediated Ca2+ Release and Mitochondrial ROS Generation Partake in the Synaptic Dysfunction Caused by Amyloid ß Peptide Oligomers.

Amyloid ß peptide oligomers (AßOs), toxic aggregates with pivotal roles in Alzheimer's disease, trigger persistent and low magnitude Ca2+ signals in neurons. We reported previously that these Ca2+ signals, which arise from Ca2+ entry and subsequent amplification by Ca2+ release through ryanodine receptor (RyR) channels, promote mitochondrial network fragmentation and reduce RyR2 expression. Here, we examined if AßOs, by inducing redox sensitive RyR-mediated Ca2+ release, stimulate mitochondrial Ca2+-uptake, ROS generation and mitochondrial fragmentation, and also investigated the effects of the antioxidant N-acetyl cysteine (NAC) and the mitochondrial antioxidant EUK-134 on AßOs-induced mitochondrial dysfunction. In addition, we studied the contribution of the RyR2 isoform to AßOs-induced Ca2+ release, mitochondrial Ca2+ uptake and fragmentation. We show here that inhibition of NADPH oxidase type-2 prevented the emergence of RyR-mediated cytoplasmic Ca2+ signals induced by AßOs in primary hippocampal neurons. Treatment with AßOs promoted mitochondrial Ca2+ uptake and increased mitochondrial superoxide and hydrogen peroxide levels; ryanodine, at concentrations that suppress RyR activity, prevented these responses. The antioxidants NAC and EUK-134 impeded the mitochondrial ROS increase induced by AßOs. Additionally, EUK-134 prevented the mitochondrial fragmentation induced by AßOs, as previously reported for NAC and ryanodine. These findings show that both antioxidants, NAC and EUK-134, prevented the Ca2+-mediated noxious effects of AßOs on mitochondrial function. Our results also indicate that Ca2+ release mediated by the RyR2 isoform causes the deleterious effects of AßOs on mitochondrial function. Knockdown of RyR2 with antisense oligonucleotides reduced by about 50% RyR2 mRNA and protein levels in primary hippocampal neurons, decreased by 40% Ca2+ release induced by the RyR agonist 4-chloro-m-cresol, and significantly reduced the cytoplasmic and mitochondrial Ca2+ signals and the mitochondrial fragmentation induced by AßOs. Based on our results, we propose that AßOs-induced Ca2+ entry and ROS generation jointly stimulate RyR2 activity, causing mitochondrial Ca2+ overload and fragmentation in a feed forward injurious cycle. The present novel findings highlight the specific participation of RyR2-mediated Ca2+ release on AßOs-induced mitochondrial malfunction.

O que pensam os universitários masculinos do nordeste brasileiro sobre sexo? / What do male university students in northeastern Brazil think about sex?

Objetivou-se identificar a percepção dos universitários masculinos nordestinos sobre o termo sexo. Estudo descritivo com abordagem qualitativa. Participaram 28 universitários masculinos acima de 18 anos, matriculados no curso de direito, através de entrevista semiestruturada, gravada, transcrita e analisadas mediante construção de duas temáticas,sexo e seus múltiplos olhares e sexo seguro, usando como referencial, Bardin. Percebeu-se que falar sobre sexo é ainda hoje considerado um tabu, visto ser difícil o diálogo aberto e direto sobre o tema, mesmo que, com todas as mudanças de comportamento, entre as famílias modernas. Mesmo com todas as transformações da sociedade e da banalidade do tem apelos meios de comunicação percebe-se que falar sobre o assunto sem timidez é algo raro. Portanto faz-se necessário que os profissionais da saúde assumam a tarefa de informar e ensinar, para que os jovens sejam sensibilizados para realização de uma prática sexual saudável.

Compreendendo as experiências vivenciadas por gestantes diabéticas durante insulinoterapia / Understanding experiences experienced by diabetic pregnant women during insulin therapy

A gestação é um período de instabilidade emocional, mudanças hormonais, e quando associada ao diagnóstico de diabetes observa-se mudança e quebra na harmonia orgânica interferindo na vida familiar e comunitária. O estudo objetivou compreender as experiências vivenciadas pelas gestantes durante a insulinoterapia. Trata-se de pesquisa com abordagem qualitativa, realizada com oito gestantes hospitalizadas por diabetes gestacional, através de entrevistas semiestruturadas numa maternidade pública, referência em atendimento a gestante de risco em Fortaleza, Ceará, nos meses de março a junho de 2013. Os depoimentos permitiram a construção de temáticas. A pesquisa obteve parecer ético nº 118/09. A descoberta do diagnóstico de diabetes gestacional revelou sentimentos e queixas da gestante em relação à necessidade de internação e tratamento com insulinoterapia. Proporcionou nas gestantes do estudo dificuldades e angústias acerca do afastamento da família; a vivência de sentimentos como medo, insegurança, além de incertezas em relação à saúde do seu filho.

Astaxanthin Protects Primary Hippocampal Neurons against Noxious Effects of Aß-Oligomers.

Increased reactive oxygen species (ROS) generation and the ensuing oxidative stress contribute to Alzheimer's disease pathology. We reported previously that amyloid-ß peptide oligomers (AßOs) produce aberrant Ca(2+) signals at sublethal concentrations and decrease the expression of type-2 ryanodine receptors (RyR2), which are crucial for hippocampal synaptic plasticity and memory. Here, we investigated whether the antioxidant agent astaxanthin (ATX) protects neurons from AßOs-induced excessive mitochondrial ROS generation, NFATc4 activation, and RyR2 mRNA downregulation. To determine mitochondrial H2O2 production or NFATc4 nuclear translocation, neurons were transfected with plasmids coding for HyperMito or NFATc4-eGFP, respectively. Primary hippocampal cultures were incubated with 0.1 µM ATX for 1.5 h prior to AßOs addition (500 nM). We found that incubation with ATX (≤10 µM) for ≤24 h was nontoxic to neurons, evaluated by the live/dead assay. Preincubation with 0.1 µM ATX also prevented the neuronal mitochondrial H2O2 generation induced within minutes of AßOs addition. Longer exposures to AßOs (6 h) promoted NFATc4-eGFP nuclear translocation and decreased RyR2 mRNA levels, evaluated by detection of the eGFP-tagged fluorescent plasmid and qPCR, respectively. Preincubation with 0.1 µM ATX prevented both effects. These results indicate that ATX protects neurons from the noxious effects of AßOs on mitochondrial ROS production, NFATc4 activation, and RyR2 gene expression downregulation.

Tecnologias do cuidado utilizadas pela enfermagem na assistência ao paciente politraumatizado: Revisão integrativa / Healthcare technologies used in nursing to care for polytraumatized patients: An integrative review / Tecnologías del cuidado utilizadas por la enfermería en la asistencia al paciente politraumatizado: Revisión integrativa

Objetivou-se identificar as tecnologias do cuidado utilizadas pelo enfermeiro na assistência ao paciente politraumatizado. Revisão integrativa, com busca de artigos em três bases de dados, no período de maio a julho de 2014. Foram selecionados 19 artigos, compreendidos no período de 2009 a 2014, distribuídos nas três categorias tecnológicas do cuidado: leves, leve-duras e duras. Verificou-se que os profissionais de enfermagem utilizam os três tipos de tecnologias do cuidado na assistência ao paciente politraumatizado, com ênfase às leve-duras. Entre as tecnologias leves: apoio e educação em saúde do paciente e familiares/cuidadores e a capacitação da equipe de enfermagem; tecnologias leve-duras: gerência do cuidado, acolhimento do paciente com classificação de risco, avaliação e tratamento da dor, processo de enfermagem e elaboração de protocolos; e tecnologias duras: sistemas de informação. Percebeu-se a melhoria assistencial proporcionada pelas tecnologias do cuidado, por estas abrangerem todos os aspectos do cuidar (AU).

The aim of this study was to identify healthcare technologies used by nurses in the care for polytraumatized patients. This is an integrative review, with articles found in three databases in the period from May to July 2014. Nineteen articles from the period between 2009 and 2014 were selected and distributed among the three categories of healthcare technology: soft, soft-hard and hard. It was found that nursing workers adopted the three types of healthcare technology when caring for polytraumatized patients, emphasizing the soft-hard category. Soft technologies included health support and education of patient and family members/caregivers and training of the nursing team; soft-hard technologies: care management, admission of patients under risk, assessment and treatment of pain, nursing process and development of protocols; and hard technologies: information systems. Care improvements caused by healthcare technologies were found, since they reach all aspects of care (AU).

El objetivo del estudio fue identificar las tecnologías del cuidado utilizadas por el enfermero en la asistencia al paciente politraumatizado. Revisión integrativa, cuya búsqueda de artículos ocurrió en tres bases de datos, en el periodo de mayo a julio de 2014. Fueron seleccionados 19 artículos, comprendidos en el periodo de 2009 a 2014, distribuidos en las tres categorías tecnológicas de cuidado: leves, leve-duras y duras. Se verificó que los profesionales de enfermería utilizan los tres tipos de tecnologías del cuidado en la asistencia al paciente politraumatizado, con énfasis a las leve-duras. Entre las tecnologías leves: apoyo y educación en salud del paciente y familiares/cuidadores y la capacitación del equipo de enfermería; tecnologías leve-duras: gerencia del cuidado, acojimiento del paciente con clasificación de riesgo, evaluación y tratamiento del dolor, proceso de enfermería y elaboración de protocolos; y tecnologías duras: sistemas de información. Se percibió la mejoría asistencial proporcionada por las tecnologías del cuidado, a causa de que estas abarcan todos los aspectos del cuidar (AU).

Adesão e permanência de mulheres no boxe em Curitiba-PR / Adhesion and permanence in women's boxing in Curitiba-PR / Adhesión y retención en el boxeo femenino en Curitiba-PR

O estudo procurou compreender quais motivações levam as mulheres a entrarem e permanecerem no boxe feminino em quatro academias de Curitiba/PR, uma vez que observa-se grande busca do sexo feminino por essa modalidade nessa realidade. Foram realizadas entrevistas semiestruturadas com 08 mulheres praticantes da modalidade há pelo menos seis meses, as quais foram analisadas de acordo com os seguintes eixos temáticos: saúde/estética, escolha do esporte, incentivo e socialização, esporte para homens. Constatou-se que os principais motivos para a adesão são a localização, sociabilidade e ambiente, e para a permanência, o alto gasto calórico (finalidades estéticas) e o dinamismo do esporte. Além de fatores que convergem para a adesão e aderência, como o ambiente e a diminuição e controle do estresse. Desta maneira, as mulheres procuram essa prática por anseios que vão muito além do boxe tradicional, e esse movimento de inserção tende a elevar-se cada vez mais.

The study sought to understand motivations which lead women to enter and remain in women's boxing in four fitness centers in Curitiba / PR, since there is great looking female of this way that reality. Were performed semi-structured interviews with 08 women practitioners of the sport were held for at least six months, which were analyzed according to the following themes: health / beauty, choice of sport, encouragement and socializing, sport for men. It was found that the main reasons for joining are the location, sociability and environment, and for the stay, the high energy expenditure (aesthetic purposes) and the dynamism of the sport. In addition to factors that converge to the adhesion and permanence, such as the environment and the reduction and control of stress. Thus, women seek this practice by anxieties that go far beyond the traditional boxing, and this insertion movement tends to rise more and more.

El estudio trata de entender qué motivaciones llevan a las mujeres a entrar y permanecer en el boxeo femenino en cuatro centros de fitness en Curitiba / PR. Se realizaron entrevistas semi-estructuradas con 08 mujeres que practican este deporte durante al menos seis meses, lo que se analizaron de acuerdo con los temas siguientes: salud / belleza, la elección del deporte, el estímulo y la socialización, el deporte para los hombres. Se encontró que las principales razones para adhésion son la ubicación, la sociabilidad y el medio ambiente, y por la retención, el alto gasto de energía (con fines estéticos) y el dinamismo del deporte. Para adhésion y retención los factores son el medio ambiente y la reducción y control del estrés. Por lo tanto, las mujeres buscan esta práctica por la ansiedad que van mucho más allá del boxeo tradicional, y este movimiento de inserción tiende a subir más.