OsRMC, a negative regulator of salt stress response in rice, is regulated by two AP2/ERF transcription factors

High salinity causes remarkable losses in rice productivity worldwide mainly because it inhibits growth and reduces grain yield. To cope with environmental changes, plants evolved several adaptive mechanisms, which involve the regulation of many stress-responsive genes. Among these, we have chosen OsRMC to study its transcriptional regulation in rice seedlings subjected to high salinity. Its transcription was highly induced by salt treatment and showed a stress-dose-dependent pattern. OsRMC encodes a receptor-like kinase described as a negative regulator of salt stress responses in rice. To investigate how OsRMC is regulated in response to high salinity, a salt-induced rice cDNA expression library was constructed and subsequently screened using the yeast one-hybrid system and the OsRMC promoter as bait. Thereby, two transcription factors (TFs), OsEREBP1 and OsEREBP2, belonging to the AP2/ERF family were identified. Both TFs were shown to bind to the same GCC-like DNA motif in OsRMC promoter and to negatively regulate its gene expression. The identified TFs were characterized regarding their gene expression under different abiotic stress conditions. This study revealed that OsEREBP1 transcript level is not significantly affected by salt, ABA or severe cold (5 °C) and is only slightly regulated by drought and moderate cold. On the other hand, the OsEREBP2 transcript level increased after cold, ABA, drought and high salinity treatments, indicating that OsEREBP2 may play a central role mediating the response to different abiotic stresses. Gene expression analysis in rice varieties with contrasting salt tolerance further suggests that OsEREBP2 is involved in salt stress response in rice.

Protection against oxidative stress through SUA7/TFIIB regulation in Saccharomyces cerevisiae

The general transcription factor TFIIB, encoded by SUA7 in Saccharomyces cerevisiae, is required for transcription activation but apparently of a specific subset of genes, for example, linked with mitochondrial activity and hence with oxidative environments. Therefore, studying SUA7/TFIIB as a potential target of oxidative stress is fundamental. We found that controlled SUA7 expression under oxidative conditions occurs at transcriptional and mRNA stability levels. Both regulatory events are associated with the transcription activator Yap1 in distinct ways: Yap1 affects SUA7 transcription up regulation in exponentially growing cells facing oxidative signals; the absence of this activator per se contributes to increase SUA7 mRNA stability. However, unlike SUA7 mRNA, TFIIB abundance is not altered on oxidative signals. The biological impact of this preferential regulation of SUA7 mRNA pool is revealed by the partial suppression of cellular oxidative sensitivity by SUA7 overexpression, and supported by the insights on the existence of a novel RNA-binding factor, acting as an oxidative sensor, which regulates mRNA stability. Taken together the results point out a primarily cellular commitment to guarantee SUA7 mRNA levels under oxidative environments.

Expression of YAP4 in Saccharomyces cerevisiae under osmotic stress

YAP4, a member of the yeast activator protein (YAP) gene family, is induced in response to osmotic shock in the yeast Saccharomyces cerevisiae. The null mutant displays mild and moderate growth sensitivity at 0.4 M and 0.8 M NaCl respectively, a fact that led us to analyse YAP4 mRNA levels in the hog1 (high osmolarity glycerol) mutant. The data obtained show a complete abolition of YAP4 gene expression in this mutant, placing YAP4 under the HOG response pathway. YAP4 overexpression not only suppresses the osmosensitivity phenotype of the yap4 mutant but also relieves that of the hog1 mutant. Induction, under the conditions tested so far, requires the presence of the transcription factor Msn2p, but not of Msn4p, as YAP4 mRNA levels are depleted by at least 75% in the msn2 mutant. This result was further substantiated by the fact that full YAP4 induction requires the two more proximal stress response elements. Furthermore we find that GCY1, encoding a putative glycerol dehydrogenase, GPP2, encoding a NAD-dependent glycerol-3-phosphate phosphatase, and DCS2, a homologue to a decapping enzyme, have decreased mRNA levels in the yap4 -deleted strain. Our data point to a possible, as yet not entirely understood, role of the YAP4 in osmotic stress response.

Crescimento de Ulva rigida C. Agardh em condições de stress de macronutrientes

Dissertação de Mestrado em Biodiversidade e Biotecnologia Vegetal

Characterization of a polyubiquitin gene in T. thermophila and of ubiquitin gene expression during sexual reproduction and under stress conditions

A 5-unit polyubiquitin gene, TTU3, was isolated from a T. thermophila genomic library and sequenced. This gene presents an extra triplet coding for Phe, a AGAGA motif and a putative HSE element in its 5'-non-coding region. The ubiquitin gene expression in this ciliate was investigated by Northern blot hybridization in conjugating cells or cells under stress conditions. Exponentially growing cells express two ubiquitin mRNAs of 0.75 and 1.8 kb and a new species of 1.4 kb is induced under hyperthermic stress. During sexual reproduction of the cells (conjugation) the 1.8-kb mRNA is still transcribed whereas the steady-state population of the 0.75 mRNA transcripts is strongly diminished. Southern blot analysis suggests that ubiquitin in T. thermophila constitutes a large family of about ten members.

Adverse effect of alcohol drinking: the role of oxidative stress

Atualmente, o álcool tem um papel importante na saúde pública e surge como um dos principais problemas sociais no mundo, dado que é a droga mais viciante aceite em encontros sociais. Provavelmente, por essa razão, os riscos do consumo abusivo do álcool são subestimados pelos jovens, mulheres grávidas e idosos. O álcool, quando ingerido em altas proporções, pode afetar todos os órgãos e desencadear inúmeras doenças, tais como a doença cardíaca coronariana, doença neurodegenerativa, as doenças crónicas e câncer. O álcool afeta ainda o estado psicológico, induzindo a violência, o estado antissocial e situações de risco de comportamentos. Por estas razões, o álcool tornou-se um foco principal da investigação, avaliando os seus efeitos sobre o corpo humano. Nesta pesquisa, foram suscitadas amostras de sangue de um grupo de pacientes em tratamento psicológico e/ou farmacêutico que serão analisadas com quatro métodos: Teste de Radicais Livres do Oxigénio (FORT), Defesa contra Radicais Livres do Oxigénio (FORD), cromatografia gasosa (GC) e cromatografia líquida de alta pressão (HPLC). Ambos os métodos FORT e FORD avaliam o stress oxidativo pela quantificação de radicais livres e a capacidade de antioxidantes em eliminar esses radicais livres, respetivamente. O stress oxidativo é o efeito do excesso de consumo de álcool, que é reduzido pela capacidade de ação dos antioxidantes. A boa reprodutibilidade, precisão e exatidão de ambos os métodos indicam que estes podem ser aplicados em rápidos diagnósticos. Para o método FORT e considerando o início do tratamento, os pacientes alcoólicos apresentaram uma média de 3,59±1.01mmol/LH2O2 e o grupo de controlo uma média de 1,42±0.53mmol/LH2O2, o que mostra uma diferença significativa entre os dois grupos (P=0,0006). Para o método FORD, pacientes alcoólicos apresentam uma média de 1,07±0.53mmol/LH2O2 e o grupo de controlo, uma média de 2,81±0.46mmol/LH2O2, mostrando também uma média significativa (P=0,0075). Após 15 dias de tratamento observou-se que há uma diferença entre os dois grupos de pacientes alcoólicos, mas não há nenhum melhoramento em relação ao grupo de pacientes em tratamento. No método FORT os grupos mostram uma diferença significativa (P=0,0073), tendo os pacientes sem tratamento farmacêutico melhores resultados (2.37±0.44mmol/LH2O2) do que os pacientes com tratamento (3.72±1,04mmol/LH2O2). O oposto ocorre no método FORD, os pacientes em tratamento farmacêutico presentam melhores resultados (1.16±0.65mmol/LH2O2) do que o outro grupo (0.75±0.22mmol/LH2O2), não sendo, no entanto, uma diferença significativa entre os dois grupos (P=0.16). Os resultados obtidos para a concentração de MDA pelo método de HPLC mostraram que o grupo de controlo tem valores mais baixos do que os pacientes alcoólicos, embora a diferença não seja muito significativa (P = 0,084), mas é ainda elevada. Além disso, os dois grupos de pacientes não apresentaram uma diferença significativa entre os seus resultados no início (P=0,77) e no fim (P=0,79) do tratamento. De acrescentar ainda que, os resultados da concentração de álcool no sangue determinados pelo método de CG mostraram que só alguns pacientes sem tratamento consumiram álcool durante o período de tratamento, o que influencia negativamente a conclusão sobre o efeito do tratamento. Contudo, outros fatores externos podem ainda influenciar os resultados finais, tais como o estado nutricional e estado psicológico dos pacientes, se o paciente continua a beber durante o tempo de tratamento ou até mesmo se o paciente é exposto a outros tipos de substâncias nocivas. Existe ainda a possibilidade de o tempo de aplicação do tratamento não ser suficiente para apresentar um efeito positivo em relação ao stress oxidativo e este é um outro fator que contribui para a impossibilidade de confirmar sobre o efeito, quer seja positivo ou negativo, do tratamento antioxidante.

Efeito de diferentes concentrações de antioxidantes na proteção contra o stress oxidativo e na viabilidade espermática durante a criopreservação do Sémen Equino

Dissertação de Mestrado, Engenharia Zootécnica, 4 de Janeiro de 2016, Universidade dos Açores.

Stress diário e saúde ocular – Corioretinopatia serosa central: estudo de caso

Objectivos - Na sociedade atual as pessoas são diariamente expostas a inúmeras situações de stress. As exaltações vividas são transformadas em energia pulsional que por sua vez deve ser despendida segundo três vias: psíquica, motora e visceral. Se esta energia não for despendida pelas duas primeiras vias, acumula-se como carga psíquica originando tensão psíquica. A tensão gerada vai repercutir-se na saúde de cada indivíduo. Uma das causas da Coriorretinopatia Central Serosa (CRCS) é o stress. A CRCS é uma patologia de carácter esporádico, caracterizada por um descolamento seroso espontâneo da retina neuro sensorial na região macular, com consequente comprometimento da acuidade visual (AV). Sendo uma patologia de carácter esporádico desencadeada por situações de stress, pretende-se caracterizar as manifestações consequentes desta patologia, bem como abordar os factores diagnósticos da mesma, no caso em estudo. Metodologia - Estudo de caso. Análise de um paciente do sexo masculino, de 36 anos de idade, com diagnóstico de CRCS. Resultados - A CRCS caracteriza-se por um descolamento seroso da retina neuro sensorial na região macular, com comprometimento da visão. O paciente chegou à consulta com queixa de baixa de acuidade visual súbita do olho esquerdo. Realizaram-se exames para concluir o diagnóstico sendo estes, OCT, angiografia e avaliação da acuidade visual. Na angiografia verifica-se afeção da área macular, ao OCT verifica-se descolamento seroso da retina neuro sensorial na região macular. A acuidade visual era de 6/10 no olho esquerdo e 10/10 no olho direito. Apresentava ainda queixas de metamorfopsias. Discussão / Conclusão - Ainda que a etiologia da CRCS não seja totalmente conhecida, encontra-se associada a situações de stress, a hipertensão arterial, a comportamentos decorrentes da forma exagerada e ansiosa de viver, à competitividade, gravidez e medicação sistémica de esteróides. O tratamento inicial recomendado é repouso absoluto, caso se verifique insucesso o tratamento passará pela realização de terapia fotodinâmica.

Sleep, stress and compensatory behaviors in Australian nurses and midwives

OBJECTIVE: To describe sleep, stress and compensatory behaviors in nurses and midwives. METHODS: The study included 41 midwives and 21 nurses working in Australian hospitals between 2005 and 2009. Participation was voluntary. All participants recorded on a daily basis their work and sleep hours, levels of stress and exhaustion, caffeine intake and use of sleep aids for a month (1,736 days, 1,002 work shifts). RESULTS: Participants reported moderate to high levels of stress and exhaustion on 20-40% of work days; experienced sleep disruption on more than 50% of work days; struggled to remain awake on 27% of work days; and suffered extreme drowsiness or experienced a near accident while travelling home on 9% of workdays. Age, perceived sleep duration and work hours were significant predictors of caffeine intake. About 60% of participants reported using sleep aids (about 20% reported taking prescription medications and 44% of nurses and 9% of midwives reported alcohol use as a sleep aid at least once during the study). Stress and workdays were significant predictors of sedative use. Overall, 22% reported being indifferent or mildly dissatisfied with their job. CONCLUSIONS: Sleep problems, high levels of stress and exhaustion and low job satisfaction are prevalent among nurses and midwives. The use of alcohol and sleeping pills as sleep aids, and the use of caffeine to help maintain alertness is also common. Nurses and midwives may use caffeine to compensate for reduced sleep, especially on workdays, and sleeping pills to cope with their daily work-related stress.

Severe stress urinary incontinence: pelvic floor muscle training program

International guidelines recommend a first line therapy in the treatment of female stress urinary incontinence (SUI), the pelvic floor muscle (PFM) training. This case report assesses the effects of the PFM training program in treating women with severe SUI. The urodynamic parameters allow diagnosed intrinsic sphincter deficiency and urethral hypermobility. The subjective and objective parameters were assessed at the beginning and after six-month of PFM training program. This case report confirms the efficiency of the intensive training program in severe SUI. The medical implications of the PFM training as first treatment option reflect favourable individual results and additionally contribute to the selection of the non-invasive treatment, the reduction of the incidence collateral effects, low costs and that does not prevent future treatment options.

Radiographers practice concerning breast compression in mammography: gradual versus no gradual

Purpose: Evaluate the type of breast compression (gradual or no gradual) that provides less discomfort to the patient. Methods and Materials: The standard projections were simulated [craniocaudal/(CC) and mediolateral-oblique/(MLO)] with the two breast compressions in 90 volunteers women aged between 19 and 86. The women were organised in groups according to the breast density. The intensity of discomfort was evaluated using the scale that have represented several faces (0-10) proposed by Wong Baker in the end of each simulation. It was also applied an interview using focus group to debate the score that were attributed during pain evaluation and to identify the criteria that were considered to do the classification. Results: The women aged between 19-29y (with higher breast density) classified the pain during no gradual compression as 4 and the gradual compression as 2 for both projections. The MLO projection was considered the most uncomfortable. During the focus group interview applied to this group was highlighted that compression did not promoted pain but discomfort. They considered that the high expectations of pain did not correspond to the discomfort that they felt. Similar results were identified for the older women (30-50y; > 50y). Conclusion: The radiographers should considerer the technique for breast compression. The gradual compression was considered for the majority of the women as the most comfortable regardless of breast density. The MLO projection was considered as uncomfortable due to the positioning (axila and inclusion of pectoral muscle) and due to the higher breast compression compared to the CC projection.

Occupational stress and coping resources in physiotherapists: a survey of physiotherapists in three general hospitals

Objectives - To identify occupational stressors and coping resources in a group of physiotherapists, and to analyse interactions between subjective levels of stress, efficacy in stress resolution and coping resources used by these professionals. Design - A sample of 55 physiotherapists working in three general hospitals in Portugal completed the Coping Resources Inventory for Stress, the Occupational Stressors Inventory and two subjective scales for stress and stress resolution. Main results - Most physiotherapists perceived that they were moderately stressed (19/55, 35%) or stressed (20/55, 36%) due to work, and reported that their efficacy in stress resolution was moderate (25/54, 46%) or efficient (23/54, 42%). Issues related to lack of professional autonomy, lack of organisation in the hierarchical command chain, lack of professional and social recognition, disorganisation in task distribution and interpersonal conflicts with superiors were identified as the main sources of stress. The most frequently used coping resources were social support, stress monitoring, physical health and structuring. Perceived efficacy in stress resolution was inversely related to perceived level of occupational stress (r = −0.61, P < 0.01). Significant correlations were found between several coping resources and the perceived level of stress and efficacy in stress resolution. Associations between problem solving, cognitive restructuring and stress monitoring and both low levels of perceived stress and high levels of perceived efficacy were particularly strong. Implications for practice - The importance of identifying stressors and coping resources related to physiotherapists’ occupational stress, and the need for the development of specific training programmes to cope with stress are supported.

Buckling strength of adhesively-bonded single and double-strap repairs on carbon-epoxy structures

This work reports on an experimental and finite element method (FEM) parametric study of adhesively-bonded single and double-strap repairs on carbon-epoxy structures under buckling unrestrained compression. The influence of the overlap length and patch thickness was evaluated. This loading gains a particular significance from the additional characteristic mechanisms of structures under compression, such as fibres microbuckling, for buckling restrained structures, or global buckling of the assembly, if no transverse restriction exists. The FEM analysis is based on the use of cohesive elements including mixed-mode criteria to simulate a cohesive fracture of the adhesive layer. Trapezoidal laws in pure modes I and II were used to account for the ductility of most structural adhesives. These laws were estimated for the adhesive used from double cantilever beam (DCB) and end-notched flexure (ENF) tests, respectively, using an inverse technique. The pure mode III cohesive law was equalled to the pure mode II one. Compression failure in the laminates was predicted using a stress-based criterion. The accurate FEM predictions open a good prospect for the reduction of the extensive experimentation in the design of carbon-epoxy repairs. Design principles were also established for these repairs under buckling.

Mechanical behavior and localized failure modes in a porous basalt from the Azores

Basaltic rocks are the main component of the oceanic upper crust, thus of potential interest for water and geothermal resources, storage of CO2 and volcanic edifice stability. In this work, we investigated experimentally the mechanical behavior and the failure modes of a porous basalt, with an initial connected porosity of 18%. Results were acquired under triaxial compression experiments at confining pressure in the range of 25-200 MPa on water saturated samples. In addition, a purely hydrostatic test was also performed to reach the pore collapse critical pressure P*. During hydrostatic loading, our results show that the permeability is highly pressure dependent, which suggests that the permeability is mainly controlled by pre-existing cracks. When the sample is deformed at pressure higher than the pore collapse pressure P*, some very small dilatancy develops due to microcracking, and an increase in permeability is observed. Under triaxial loading, two modes of deformation can be highlighted. At low confining pressure (Pc < 50 MPa), the samples are brittle and shear localization occurs. For confining pressure > 50 MPa, the stress-strain curves are characterized by strain hardening and volumetric compaction. Stress drops are also observed, suggesting that compaction may be localized. The presence of compaction bands is confirmed by our microstructure analysis. In addition, the mechanical data allows us to plot the full yield surface for this porous basalt, which follows an elliptic cap as previously observed in high porosity sandstones and limestones.

The influence of geometric factors on the wall shear stress distribution in realistic human coronary arteries

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia Biomédica. A presente dissertação foi desenvolvida no Erasmus Medical Center em Roterdão, Holanda