Oscillating Aquaporin Phosphorylation and 14-3-3 Proteins Mediate the Circadian Regulation of Leaf Hydraulics.

The circadian clock regulates plant tissue hydraulics to synchronize water supply with environmental cycles and thereby optimize growth. The circadian fluctuations in aquaporin transcript abundance suggest that aquaporin water channels play a role in these processes. Here, we show that hydraulic conductivity (K ros) of Arabidopsis (Arabidopsis thaliana) rosettes displays a genuine circadian rhythmicity with a peak around midday. Combined immunological and proteomic approaches revealed that phosphorylation at two C-terminal sites (Ser280, Ser283) of PLASMA MEMBRANE INTRINSIC PROTEIN 2;1 (AtPIP2;1), a major plasma membrane aquaporin in rosettes, shows circadian oscillations and is correlated with K ros Transgenic expression of phosphodeficient and phosphomimetic forms of this aquaporin indicated that AtPIP2;1 phosphorylation is necessary but not sufficient for K ros regulation. We investigated the supporting role of 14-3-3 proteins, which are known to interact with and regulate phosphorylated proteins. Individual knockout plants for five 14-3-3 protein isoforms expressed in rosettes lacked circadian activation of K ros Two of these [GRF4 (14-3-3Phi); GRF10 (14-3-3Epsilon)] showed direct interactions with AtPIP2;1 in the plant and upon coexpression in Xenopus laevis oocytes and activated AtPIP2;1, preferentially when the latter was phosphorylated at its two C-terminal sites. We propose that this regulatory mechanism assists in the activation of phosphorylated AtPIP2;1 during circadian regulation of K ros.

Coordinating light responses between the nucleus and the chloroplast, a role for plant cryptochromes and phytochromes.

To promote photomorphogenesis, including plastid development and metabolism, the phytochrome (phy) and the cryptochrome (cry) photoreceptors orchestrate genome-wide changes in gene expression in response to Red (R)- and Blue (B)-light cues. While phys and crys have a clear role in modulating photosynthesis, their role in the coordination of the nuclear genome and the plastome, essential for functional chloroplasts, remains underexplored. Using publicly available genome datasets for WT and phyABCDE or cry1cry2 Arabidopsis seedlings, grown, respectively, under R- or B-light, we bioinformatically analyzed the influence of light inputs and photoreceptors in the control of nuclear genes with a function in the chloroplast, and evaluated the role of phyB in the modulation of plastome-encoded genes. We show gene co-induction by R-phys and B-crys for genes with a chloroplastic function, and also apparent photoreceptor-driven preferential responses. Evidence from phyB in Arabidopsis together with published evidence from CRY2 in tomato also supports the participation of both photoreceptor families in the global modulation of the plastome genes. To begin addressing how these light-sensors orchestrate changes in an organellar genome, we evaluated their effect over genes with potential functions in plastid gene-expression regulation based on their TAIR annotation. Results indicate that both crys and phys modulate 'plastome-regulatory genes' with enrichment in the contribution of crys to all processes and of phys to post-transcription and transcription. Furthermore, we identified a new role for HY5 as a relevant light-signaling component in photoreceptor-based anterograde signaling leading to plastome gene regulation.

Regulation of Arabidopsis leaf hydraulics involves light-dependent phosphorylation of aquaporins in veins.

The water status of plant leaves depends on the efficiency of the water supply, from the vasculature to inner tissues. This process is under hormonal and environmental regulation and involves aquaporin water channels. In Arabidopsis thaliana, the rosette hydraulic conductivity (Kros) is higher in darkness than it is during the day. Knockout plants showed that three plasma membrane intrinsic proteins (PIPs) sharing expression in veins (PIP1;2, PIP2;1, and PIP2;6) contribute to rosette water transport, and PIP2;1 can fully account for Kros responsiveness to darkness. Directed expression of PIP2;1 in veins of a pip2;1 mutant was sufficient to restore Kros. In addition, a positive correlation, in both wild-type and PIP2;1-overexpressing plants, was found between Kros and the osmotic water permeability of protoplasts from the veins but not from the mesophyll. Thus, living cells in veins form a major hydraulic resistance in leaves. Quantitative proteomic analyses showed that light-dependent regulation of Kros is linked to diphosphorylation of PIP2;1 at Ser-280 and Ser-283. Expression in pip2;1 of phosphomimetic and phosphorylation-deficient forms of PIP2;1 demonstrated that phosphorylation at these two sites is necessary for Kros enhancement under darkness. These findings establish how regulation of a single aquaporin isoform in leaf veins critically determines leaf hydraulics.

CPK13, a noncanonical Ca2+-dependent protein kinase, specifically inhibits KAT2 and KAT1 shaker K+ channels and reduces stomatal opening.

Ca(2) (+)-dependent protein kinases (CPKs) form a large family of 34 genes in Arabidopsis (Arabidopsis thaliana). Based on their dependence on Ca(2+), CPKs can be sorted into three types: strictly Ca(2+)-dependent CPKs, Ca(2+)-stimulated CPKs (with a significant basal activity in the absence of Ca(2+)), and essentially calcium-insensitive CPKs. Here, we report on the third type of CPK, CPK13, which is expressed in guard cells but whose role is still unknown. We confirm the expression of CPK13 in Arabidopsis guard cells, and we show that its overexpression inhibits light-induced stomatal opening. We combine several approaches to identify a guard cell-expressed target. We provide evidence that CPK13 (1) specifically phosphorylates peptide arrays featuring Arabidopsis K(+) Channel KAT2 and KAT1 polypeptides, (2) inhibits KAT2 and/or KAT1 when expressed in Xenopus laevis oocytes, and (3) closely interacts in plant cells with KAT2 channels (Förster resonance energy transfer-fluorescence lifetime imaging microscopy). We propose that CPK13 reduces stomatal aperture through its inhibition of the guard cell-expressed KAT2 and KAT1 channels.

Coordinated post-translational responses of aquaporins to abiotic and nutritional stimuli in Arabidopsis roots.

In plants, aquaporins play a crucial role in regulating root water transport in response to environmental and physiological cues. Controls achieved at the post-translational level are thought to be of critical importance for regulating aquaporin function. To investigate the general molecular mechanisms involved, we performed, using the model species Arabidopsis, a comprehensive proteomic analysis of root aquaporins in a large set of physiological contexts. We identified nine physiological treatments that modulate root hydraulics in time frames of minutes (NO and H2O2 treatments), hours (mannitol and NaCl treatments, exposure to darkness and reversal with sucrose, phosphate supply to phosphate-starved roots), or days (phosphate or nitrogen starvation). All treatments induced inhibition of root water transport except for sucrose supply to dark-grown plants and phosphate resupply to phosphate-starved plants, which had opposing effects. Using a robust label-free quantitative proteomic methodology, we identified 12 of 13 plasma membrane intrinsic protein (PIP) aquaporin isoforms, 4 of the 10 tonoplast intrinsic protein isoforms, and a diversity of post-translational modifications including phosphorylation, methylation, deamidation, and acetylation. A total of 55 aquaporin peptides displayed significant changes after treatments and enabled the identification of specific and as yet unknown patterns of response to stimuli. The data show that the regulation of PIP and tonoplast intrinsic protein abundance was involved in response to a few treatments (i.e. NaCl, NO, and nitrate starvation), whereas changes in the phosphorylation status of PIP aquaporins were positively correlated to changes in root hydraulic conductivity in the whole set of treatments. The identification of in vivo deamidated forms of aquaporins and their stimulus-induced changes in abundance may reflect a new mechanism of aquaporin regulation. The overall work provides deep insights into the in vivo post-translational events triggered by environmental constraints and their possible role in regulating plant water status.

Política nacional de humanização nas ações do acolhimento dos profissionais de saúde em unidade de terapia intensiva / National humanization policy in welcoming healthcare professionals in intensive care units

Atualmente a humanização é ponto de pauta nos encontros nacionais e internacionais da área da saúde, assim, cada vez mais enfermeiros nas Unidades de Terapia Intensiva têm se preoucupado com sua prática e implementação em suas unidades. Considerando esta preoucupação, houve através das políticas públicas, a necessidade de promover um ambiente que proporcionasse melhores condições de bem-estar, integridade física e mental do paciente e também de seus familiares. O presente estudo foi desenvolvido com o objetivo de descrever a implementação da politica nacional de humanização nas ações do acolhimento, avaliando fatores que interferem para que uma efetiva humanização da assistência. O propósito é proporcionar, de forma humanizada, acolhimento ao paciente e também á sua família nesses ambientes. Para o desenvolvimento do tema foi escolhida como forma de trabalho de revisão bibliográfica sistemática. Foi feito levantamento de artigos publicados na base de dados da Biblioteca Virtual de Saúde (BVS), utilizando-se os descritores "Enfermagem", "Terapia intensiva" e "Humanização" no período de 2003 a 2013. Ao final do levantamento, obteve-se um total de dez artigos sendo que destes, sete foram analisados por satisfazerem aos critérios de inclusão estabelecidos. Os fatores encontrados estavam relacionados ao paciente e seus familiares, a questões da equipe de enfermagem e a questões estruturais das instituições de saúde, revelando o caráter multidimensional da humanização. Conclui-se que a humanização é um aspecto fundamental aos seres humanos. Portanto, não deve apenas estar na pauta os discursos, mas efetivamente ser aplicada no contexto de trabalho das ações/intervenções de enfermagem. Para tanto, embora haja vários fatores interferindo, é preciso compreender as ações em várias esferas para resolvê-los.(AU)

Currently humanization is point agenda in national and international meetings of health, thus increasing nurses in Intensive Care Units have been worried for its practice and implementation in their units. Considering this preoucupação, was through public policies, the need to promote an environment that would provide better welfare, physical and mental integrity of the patient and also their families. The presesnte study was developed with the aim of describing the implementation of the national policy of humanization in the actions of the host, assessing factors that influence that an effective quality care. The purpose is to provide, in a humane way, welcome to the patient and also to her family in these environments. To develop the theme was chosen as a way to work a systematic literature review. A survey was done of papers published in the database of the Virtual Health Library (VHL), using the key words "Nursing", "Intensive care" and "Humanization" in the period 2003-2013. At the end of the survey, we obtained a total of ten items and of these, seven were analyzed to meet the established criteria for inclusion. The factors found were related to the patient and their family, the issues of the nursing staff and the structural issues of health institutions, revealing the multidimensional nature of humanization. We conclude that humanization is a key aspect to humans. So it should not just be on the agenda of the talks, but effectively be applied in the context of labor actions / nursing interventions. Therefore, although there are several factors interfering, it takes compeender actions in various spheres to solve them.(AU)

La humanización está actualmente en la agenda de las reuniones nacionales e internacionales de salud, por lo que cada vez más enfermeras de las Unidades de Cuidados Intensivos se han preocupado por su práctica e implementación en sus unidades. Considerando esta preocupación, las políticas públicas han hecho necesario promover un ambiente que proporcione mejores condiciones para el bienestar y la integridad física y mental de los pacientes y sus familiares. Este estudio se realizó con el objetivo de describir la implementación de la política nacional de humanización en las áreas de acogida, evaluando los factores que interfieren en la humanización efectiva de la atención. El objetivo es proporcionar una acogida humanizada a los pacientes y sus familias en estos entornos. Para desarrollar el tema, se optó por una revisión bibliográfica sistemática. Se realizó una encuesta de artículos publicados en la base de datos de la Biblioteca Virtual en Salud (BVS), utilizando los descriptores "Enfermería", "Cuidados Intensivos" y "Humanización" de 2003 a 2013. Al final de la encuesta, se obtuvieron un total de diez artículos, siete de los cuales fueron analizados por cumplir los criterios de inclusión establecidos. Los factores encontrados estaban relacionados con los pacientes y sus familias, cuestiones del personal de enfermería y cuestiones estructurales de las instituciones sanitarias, lo que revela el carácter multidimensional de la humanización. La conclusión es que la humanización es un aspecto fundamental para el ser humano. Por lo tanto, no debe figurar únicamente en el orden del día de los discursos, sino aplicarse efectivamente en el contexto de las acciones/intervenciones de enfermería. Para ello, aunque existan varios factores que interfieren, es necesario comprender las acciones en diversas esferas para resolverlos.(AU)

Auditoria de contas: impacto de glosas ocorrido a falta de anotações de enfermagem / Audit of accounts: impact of glosses due to lack of nursing notes

A auditoria foi instituída no sistema de saúde para organizar, planejar e direcionar os recursos financeiros, tendo como maior parte das glosas hospitalares, justificada por ausência de anotações, principalmente ações das equipes de enfermagem e médica. É importante lembrar que os registros de enfermagem estão vinculados à grande parte do pagamento de materiais, medicamentos e procedimentos, principais fontes de lucratividade das instituições hospitalares. Sendo Assim, o principal meio de assegurar o recebimento do valor gasto é através das corretas anotações de enfermagem. Este estudo teve como objetivo de realizar um levantamento bibliográfico, sobre os impactos causado pelo registro de enfermagem referente as glosas hospitalares. Trata-se de uma pesquisa de revisão integrativa, classifica-se como qualitativa, do tipo exploratória e retrospectiva, foram avaliados vários artigos publicados com a temática sobre anotações de enfermagem e glosas. Conforme análise dos artigos selecionados para o estudo, evidenciou-se que existe a falta de anotações, checagem e carimbo por parte da equipe, refletindo em glosas durante o processo de faturamento hospitalar.(AU)

The audit was instituted in the health system to organize, plan and direct financial re-sources, with most of the hospital glosses, justified by the absence of notes, mainly ac-tions of the nursing and medical teams. It is important to remember that nursing records are linked to a large part of the payment for materials, medications and procedures, the main sources of profitability for hospital institutions. Therefore, the main means of ensur-ing receipt of the amount spent is through the correct nursing notes. This study aimed to carry out a bibliographic survey on the impacts caused by the nursing record referring to hospital glosses. This is an integrative review research, it is classified as qualitative, ex-ploratory and retrospective, several articles published with the theme of nursing notes and glosses were evaluated. According to the analysis of the articles selected for the study, it was evidenced that there is a lack of notes, checking and stamping by the team, reflecting in glosses during the hospital billing process.(AU)

La auditoría fue instituida en el sistema de salud para organizar, planificar y direccionar los recursos financieros, con la mayor parte de las glosas hospitalarias, justificadas por la ausencia de notas, principalmente de las acciones de los equipos de enfermería y médicos. Es importante recordar que los registros de enfermería están vinculados a gran parte del pago de materiales, medicamentos y procedimientos, principales fuentes de rentabilidad de las instituciones hospitalarias. Por lo tanto, el principal medio de garantizar la recepción del importe gastado es a través de las notas de enfermería correctas. Este estudio tuvo como objetivo realizar una pesquisa bibliográfica sobre los impactos causados por el registro de enfermería referente a las glosas hospitalarias. Se trata de una investigación de revisión integradora, se clasifica como cualitativa, ex-ploratoria y retrospectiva, se evaluaron varios artículos publicados con el tema de notas de enfermería y glosas. De acuerdo con el análisis de los artículos seleccionados para el estudio, se evidenció la falta de anotaciones, verificación y sellado por parte del equipo, reflejándose en glosas durante el proceso de facturación hospitalaria.(AU)

The Arabidopsis guard cell outward potassium channel GORK is regulated by CPK33.

A complex signaling network involving voltage-gated potassium channels from the Shaker family contributes to the regulation of stomatal aperture. Several kinases and phosphatases have been shown to be crucial for ABA-dependent regulation of the ion transporters. To date, the Ca2+ -dependent regulation of Shaker channels by Ca2+ -dependent protein kinases (CPKs) is still elusive. A functional screen in Xenopus oocytes was launched to identify such CPKs able to regulate the three main guard cell Shaker channels KAT1, KAT2, and GORK. Seven guard cell CPKs were tested and multiple CPK/Shaker couples were identified. Further work on CPK33 indicates that GORK activity is enhanced by CPK33 and unaffected by a nonfunctional CPK33 (CPK33-K102M). Furthermore, Ca2+ -induced stomatal closure is impaired in two cpk33 mutant plants.

Regulation of leaf hydraulics: from molecular to whole plant levels.

The water status of plant leaves is dependent on both stomatal regulation and water supply from the vasculature to inner tissues. The present review addresses the multiple physiological and mechanistic facets of the latter process. Inner leaf tissues contribute to at least a third of the whole resistance to water flow within the plant. Physiological studies indicated that leaf hydraulic conductance (K leaf) is highly dependent on the anatomy, development and age of the leaf and can vary rapidly in response to physiological or environmental factors such as leaf hydration, light, temperature, or nutrient supply. Differences in venation pattern provide a basis for variations in K leaf during development and between species. On a short time (hour) scale, the hydraulic resistance of the vessels can be influenced by transpiration-induced cavitations, wall collapses, and changes in xylem sap composition. The extravascular compartment includes all living tissues (xylem parenchyma, bundle sheath, and mesophyll) that transport water from xylem vessels to substomatal chambers. Pharmacological inhibition and reverse genetics studies have shown that this compartment involves water channel proteins called aquaporins (AQPs) that facilitate water transport across cell membranes. In many plant species, AQPs are present in all leaf tissues with a preferential expression in the vascular bundles. The various mechanisms that allow adjustment of K leaf to specific environmental conditions include transcriptional regulation of AQPs and changes in their abundance, trafficking, and intrinsic activity. Finally, the hydraulics of inner leaf tissues can have a strong impact on the dynamic responses of leaf water potential and stomata, and as a consequence on plant carbon economy and leaf expansion growth. The manipulation of these functions could help optimize the entire plant performance and its adaptation to extreme conditions over short and long time scales.