Interaction Between ABA Signaling and Copper Homeostasis in Arabidopsis thaliana.

ABA is involved in plant responses to non-optimal environmental conditions, including nutrient availability. Since copper (Cu) is a very important micronutrient, unraveling how ABA affects Cu uptake and distribution is relevant to ensure adequate Cu nutrition in plants subjected to stress conditions. Inversely, knowledge about how the plant nutritional status can interfere with ABA biosynthesis and signaling mechanisms is necessary to optimize stress tolerance in horticultural crops. Here the reciprocal influence between ABA and Cu content was addressed by using knockout mutants and overexpressing transgenic plants of high affinity plasma membrane Cu transporters (pmCOPT) with altered Cu uptake. Exogenous ABA inhibited pmCOPT expression and drastically modified COPT2-driven localization in roots. ABA regulated SPL7, the main transcription factor responsive for Cu deficiency responses, and subsequently affected expression of its targets. ABA biosynthesis (aba2) and signaling (hab1-1 abi1-2) mutants differentially responded to ABA according to Cu levels. Alteration of Cu homeostasis in the pmCOPT mutants affected ABA biosynthesis, transport and signaling as genes such as NCED3, WRKY40, HY5 and ABI5 were differentially modulated by Cu status, and also in the pmCOPT and ABA mutants. Altered Cu uptake resulted in modified plant sensitivity to salt-mediated increases in endogenous ABA. The overall results provide evidence for reciprocal cross-talk between Cu status and ABA metabolism and signaling.

Modulation of copper deficiency responses by diurnal and circadian rhythms in Arabidopsis thaliana.

Copper homeostasis under deficiency is regulated by the SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7) transcription factor. The daily oscillating expression of two SPL7-dependent copper deficiency markers, COPPER TRANSPORTER (COPT2) and IRON SUPEROXIDE DISMUTASE (FSD1), has been followed by quantitative PCR and in promoter:LUCIFERASE transgenic plants. Both genes showed circadian and diurnal regulation. Under copper deficiency, their expression decreased drastically in continuous darkness. Accordingly, total copper content was slightly reduced in etiolated seedlings under copper deficiency. The expression of SPL7 and its targets COPT2 and FSD1 was differently regulated in various light signalling mutants. On the other hand, increased copper levels reduced the amplitude of nuclear circadian clock components, such as GIGANTEA (GI). The alteration of copper homeostasis in the COPT1 overexpression line and spl7 mutants also modified the amplitude of a classical clock output, namely the circadian oscillation of cotyledon movements. In the spl7 mutant, the period of the oscillation remained constant. These results suggest a feedback of copper transport on the circadian clock and the integration of rhythmic copper homeostasis into the central oscillator of plants.

Defective copper transport in the copt5 mutant affects cadmium tolerance.

Cadmium toxicity interferes with essential metal homeostasis, which is a problem for both plant nutrition and the consumption of healthy food by humans. Copper uptake is performed by the members of the Arabidopsis high affinity copper transporter (COPT) family. One of the members, COPT5, is involved in copper recycling from the vacuole toward the cytosolic compartment. We show herein that copt5 mutants are more sensitive to cadmium stress than wild-type plants, as indicated by reduced growth. Exacerbated cadmium toxicity in copt5 mutants is due specifically to altered copper traffic through the COPT5 transporter. Three different processes which have been shown to affect cadmium tolerance are altered in copt5 mutants. First, ethylene biosynthesis diminishes under copper deficiency and, in the presence of cadmium, ethylene production diminishes further. Copper deficiency responses are also attenuated under cadmium treatment. Remarkably, while copt5 roots present higher oxidative stress toxicity symptoms than controls, aerial copt5 parts display lower oxidative stress, as seen by reduced cadmium delivery to shoots. Taken together, these results demonstrate that copper transport plays a key role in cadmium resistance, and suggest that oxidative stress triggers an NADPH oxidase-mediated signaling pathway, which contributes to cadmium translocation and basal plant resistance. The slightly lower cadmium levels that reach aerial parts in the copt5 mutants, irrespective of the copper content in the media, suggest a new biotechnological approach to minimize toxic cadmium entry into food chains.

Efficacy of Artistic Actions in Raising Awareness of Gender Equality and Sexual Diversity in University Contexts.

The university community faces problems of gender inequality and intolerance toward sexual diversity that are based on heteronormativity and machismo and particularly affect women and LGTBI individuals. This study proposes that, through arts-based public pedagogy, it is possible to increase the sensitivity of the university community (PDI, PAS, and students) to gender equality and sexual diversity. This research evaluates the efficacy of a series of artistic actions carried out in the public space of three Spanish universities (UV, UPV, and UMH), based on the participants' self-assessments of their attitudes before and after witnessing these actions. Differences are analyzed by gender, university collective, and age. The results show that artistic actions have significant positive effects on the awareness of the members of the university community at cognitive, affective, and moral levels. This influence varies somewhat depending on the gender, age, and university collective. Implications of these differences are discussed, as well as the relevance of opportunities to gain knowledge in explaining them. Finally, keys to future awareness campaigns are presented.

Copper uptake mechanism of Arabidopsis thaliana high-affinity COPT transporters.

Copper (Cu) is an essential plant micronutrient. Under scarcity, Cu2+ is reduced to Cu+ and taken up through specific high-affinity transporters (COPTs). In Arabidopsis, the COPT family consists of six members, either located at the plasma membrane (COPT1, COPT2, and COPT6) or in internal membranes (COPT3 and COPT5). Cu uptake by COPT proteins has been mainly assessed through complementation studies in corresponding yeast mutants, but the mechanism of this transport has not been elucidated. To test whether Cu is incorporated by an electrogenic mechanism, electrophysiological changes induced by Cu addition were studied in Arabidopsis thaliana. Mutant (T-DNA insertion mutants, copt2-1 and copt5-2) and overexpressing lines (COPT1OE and COPT5OE) with altered expression of COPT transporters were compared to wild-type plants. No significant changes of the membrane potential (Em) were detected, regardless of genotype or Cu concentration supplied. In contrast, membrane depolarization was detected in response to iron supply in both wild-type and in mutant or transgenic plants. Similar results were obtained for trans-plant potentials (TPP). GFP fusions of the plasma membrane COPT2 and the internal COPT5 transporters were expressed in Xenopus laevis oocytes to potentiate Cu uptake signals, and the cRNA-injected oocytes were tested for electrical currents upon Cu addition using two-electrode voltage clamp. Results with oocytes confirmed those obtained in plants. Cu accumulation in injected oocytes was measured by ICP-OES, and a significant increase in Cu content with respect to controls occurred in oocytes expressing COPT2:GFP. The possible mechanisms driving this transport are discussed in this manuscript.

Ni2+ toxicity in rice: effect on membrane functionality and plant water content.

The heavy metal nickel is an essential mineral trace nutrient found at low concentrations in most natural soils. However, it may reach toxic levels in certain areas and affect a number of biochemical and physiological processes in plants. Wilting and leaf necrosis have been described as typical visible symptoms of Ni(2+) toxicity. The plasma membrane (PM) of root cells constitutes the first barrier for the entry of heavy metals but also a target of their toxic action. This work studies the relationship between disturbances of membrane functionality and the development of the typical symptoms of Ni(2+) toxicity. Rice plants (Oryza sativa L. cv. Bahia) grown in nutrient medium containing 0.5mM Ni(2+) showed a significant decrease in water content as a consequence of the stress. Addition of Ni(2+) to the solution bathing the roots induced a concentration-dependent PM depolarization but the activity of the PM-H(+)-ATPase was not inhibited by the presence of Ni(2+) and the initial resting potential recovered in less than 1h. In the short term (hours), membrane permeability of root cells was not significantly affected by Ni(2+) treatments. However, in the long term (days) a drastic loss of K(+) was measured in roots and shoots, which should be responsible for the changes in the water content measured, since stomatal conductance and the transpiration rate remained unaffected by Ni(2+) treatment. The effects induced by Ni(2+) were not permanent and could be reverted, at least in part, by transferring the plants to a medium without Ni(2+).

[Interactive patterns detection in family communication with adolescents]. / Detección de patrones interactivos en la comunicación de familias con hijos adolescentes.

Interactive patterns detection in family communication with adolescents. Nondistant communication is a relevant indicator for family functionality valuation. The goal of this study is to analyze this communication in order to identify specific kinds of leadership, interaction patterns and the relation between verbal and nonverbal elements in communication. The observational design exposed is an idiographic one, punctual and multidimensional, which uses field format as observation instrument. Participants were seven standardized families made up of both ancestors and an adolescent son or daughter. According to the family models analyzed, results show a predominantly democratic communication style in adults with recurrent support expressions. The sequential analysis incorporates only categories from the emitter point of view, and detects relevant sequences which show symmetric interaction between all three family members. Verbal and nonverbal channels provide complementary information. Depending on adolescents' gender different patterns in behaviour can be identified as well.

Daily rhythmicity of high affinity copper transport.

A differential demand for copper (Cu) of essential cupro-proteins that act within the mitochondrial and chloroplastal electronic transport chains occurs along the daily light/dark cycles. This requires a fine-tuned spatiotemporal regulation of Cu delivery, becoming especially relevant under non-optimal growth conditions. When scarce, Cu is imported through plasma membrane-bound high affinity Cu transporters (COPTs) whose coding genes are transcriptionally induced by the SPL7 transcription factor. Temporal homeostatic mechanisms are evidenced by the presence of multiple light- and clock-responsive regulatory cis elements in the promoters of both SPL7 and its COPT targets. A model is presented here for such temporal regulation that is based on the synchrony between the basal oscillatory pattern of SPL7 and its targets, such as COPT2. Conversely, Cu feeds back to coordinate intracellular Cu availability on the SPL7-dependent regulation of further Cu acquisition. This occurs via regulation at COPT transporters. Moreover, exogenous Cu affects several circadian-clock components, such as the timing of GIGANTEA transcript abundance. Together we propose that there is a dynamic response to Cu that is integrated over diurnal time to maximize metabolic efficiency under challenging conditions.

A relationship between tolerance to dehydration of rice cell lines and ability for ABA synthesis under stress.

Plant dehydration is commonly caused by some adverse environmental conditions such as salinity, drought and freezing. As the plant hormone abscisic acid (ABA) is involved in responses to water stress, we studied its putative relationship with the degree of tolerance to these abiotic stresses. For this purpose we used cell lines that had been established from mature embryos of rice (Oryza sativa L, cvs. Bahia and Bomba), and selected by their high (L-T) or low (L-S) levels of tolerance to each type of stress. Tolerance of rice calli to either osmotic, saline, or freezing stress was generally improved by a previous treatment with ABA. This ABA effect was evident in those callus lines with low tolerance (L-S), as their ability to recover from stress increased up to three fold. Independent of the cultivar used, there were no significant differences in the endogenous ABA contents between untreated L-T and L-S lines. However, upon stress, the increase in endogenous ABA was higher in L-T than in L-S lines. These results, together with those obtained by using Fluridone, an inhibitor of ABA synthesis, show that differences in the level of cell tolerance to osmotic, saline and freezing stress are related to their different capacity of ABA synthesis under stress conditions.

Temporal aspects of copper homeostasis and its crosstalk with hormones.

To cope with the dual nature of copper as being essential and toxic for cells, plants temporarily adapt the expression of copper homeostasis components to assure its delivery to cuproproteins while avoiding the interference of potential oxidative damage derived from both copper uptake and photosynthetic reactions during light hours. The circadian clock participates in the temporal organization of coordination of plant nutrition adapting metabolic responses to the daily oscillations. This timely control improves plant fitness and reproduction and holds biotechnological potential to drive increased crop yields. Hormonal pathways, including those of abscisic acid, gibberellins, ethylene, auxins, and jasmonates are also under direct clock and light control, both in mono and dicotyledons. In this review, we focus on copper transport in Arabidopsis thaliana and Oryza sativa and the presumable role of hormones in metal homeostasis matching nutrient availability to growth requirements and preventing metal toxicity. The presence of putative hormone-dependent regulatory elements in the promoters of copper transporters genes suggests hormonal regulation to match special copper requirements during plant development. Spatial and temporal processes that can be affected by hormones include the regulation of copper uptake into roots, intracellular trafficking and compartmentalization, and long-distance transport to developing vegetative and reproductive tissues. In turn, hormone biosynthesis and signaling are also influenced by copper availability, which suggests reciprocal regulation subjected to temporal control by the central oscillator of the circadian clock. This transcriptional regulatory network, coordinates environmental and hormonal signaling with developmental pathways to allow enhanced micronutrient acquisition efficiency.

Is ABA involved in tolerance responses to salinity by affecting cytoplasm ion homeostasis in rice cell lines?

The ability of plant cells to maintain cytoplasm ion homeostasis under saline stress is among the main mechanisms involved in salt tolerance. To cope with excess Na(+), cells extrude it from the cytoplasm, which requires expenditure of metabolic energy, provided by H(+) gradients generated by membrane-bound H(+)-pumps. ABA is well-known to be involved in physiological processes elicited or enhanced by stresses causing cell dehydration. In this work we studied the possible implication of this plant hormone in the control of salt-induced cellular mechanisms conducting to Na(+) extrusion from the cytoplasm. We used rice (Oryza sativa L.) cell lines selected for their different tolerance to salinity to measure the response to ABA of H(+)-pumps and Na(+)/H(+)-antiporters associated to the plasma membrane and the tonoplast. Our results show that ABA generally enhances H(+)-pumping under salt stress but not under control conditions. This effect occurs to a higher extent across the tonoplast in the more tolerant lines (L-T). Na(+)/H(+) antiport activity is practically undetectable in calli under control conditions, pre-treated or not with ABA, but shows a strong activation under salinity across the tonoplast, particularly in L-T lines (cv Bahia) and also across de plasma membrane in cv Bomba. In these lines, prior treatments with ABA tend to reduce the NaCl enhanced activity of both antiporters. Overall, under saline conditions ABA seems to affect synergistically H(+) pumping and antagonistically Na(+) extrusion. A complex network of positive and negative regulatory signals seems involved in restoring ion cell homeostasis under salt stress.

Differential salinity-induced variations in the activity of H⁺-pumps and Na⁺/H⁺ antiporters that are involved in cytoplasm ion homeostasis as a function of genotype and tolerance level in rice cell lines.

The characterisation of cellular responses to salinity in staple crops is necessary for the reliable identification of physiological markers of salinity tolerance. Under saline conditions, variations in proton gradients that are generated by membrane-bound H⁺ pumps are crucial for maintaining cytoplasm homeostasis. We examined short (15 h) and longer term effects (4 days) of NaCl stress on the H⁺ pumping activities that are associated with the plasma membrane (P-ATPase) and the tonoplast (V-ATPase and V-PPase) in rice (Oryza sativa L.) callus lines that displayed different levels of NaCl tolerance and were established from two japonica rice cultivars. The applied stress conditions were based on those that were used in the induction of a stress-responsive polyubiquitin gene promoter (UBI1) in transgenic rice calli. The most remarkable effect of NaCl stress on H⁺ pumping was the rapid activation of tonoplast-bound pumps; this was particularly observed in cv. Bomba, in which the response of the P-ATPase was slower and showed a higher level of activity after 4 days of stress. The responses were cultivar-dependent; however, in general, a stronger activation occurred in the lines that had a higher tolerance (L-T) than in the less-tolerant (L-S) lines. Substrate hydrolysis was less affected than H⁺ pumping, and it yielded higher H⁺/substrate coupling ratios, which is indicative of an enhanced H⁺ pumping efficiency under saline conditions. The Na⁺/H⁺ antiport activity was generally limited to salt-stressed calli, and higher values and stronger activation of the tonoplast antiporter were observed in the L-T lines than in the L-S lines. The results that were obtained with the NaCl-stressed transgenic lines confirmed the close relationship between metabolic activity, H⁺ pumping and the induction of Na⁺/H⁺ exchange activities.

Reducción en masa de una hernia inguinal incarcerada. Abordaje urgente por laparoscopia / En masse reduction of an incarcerated inguinal hernia. Emergency laparoscopic management

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Distinctive phytotoxic effects of Cd and Ni on membrane functionality.

Metal ions essential for plant growth, such as Fe, Mn, Ni, Cu or Zn, are taken up by plants from the soil solution through metal transporters at the plasma membrane, mainly of the ZIP and Nramp families. These transport systems, however, can also give entry to other metals (Al, Cd, Hg, Pb). Non-nutritive elements, as well as the essential nutrients at higher than metabolic concentrations, can cause phytotoxicity. We have studied previously the effects of an essential (Ni) and a non essential (Cd) heavy metal on root cell plasma membranes, the first selective barrier encountered when entering the plant, using rice as model plant. Distinctive effects of Cd and Ni on membrane function (i.e., Em and membrane permeability) were observed in the short term. We have now confirmed the pattern of Em changes caused by Cd and Ni using barley roots and have also followed the effects of both metals in longer term in rice. Our data indicate that the distinct effects caused by Cd and Ni are due to differences in cellular responses, triggered when entering the cytoplasm (i.e., an efficient detoxifying mechanism for Cd), more than to different direct effects on membranes.

[Computerized physician order entry of pharmacotherapeutic protocols in a colorectal surgery unit]. / Prescripción electrónica asistida de protocolos farmacoterapéuticos en una unidad de cirugía coloproctológica.

INTRODUCTION: Computerized physician order entry was introduced in our hospital. We present the pharmacotherapeutic protocols that we use in patient care. MATERIAL AND METHODS: Six pharmaceutical protocols were designed by consensus in a colorectal surgery unit and were applied over a 2-year period. Patients undergoing ambulatory or urgent surgery were excluded. RESULTS: We treated 772 patients. Two hundred twenty patients (28.5%) were assigned to the preoperative protocol for minor proctologic surgery. After surgery, all 220 patients were included in the postoperative protocol for minor proctologic surgery. The remaining 552 patients (71.5%) were assigned to the protocol for major colorectal surgery. The preoperative protocol for major colorectal surgery was indicated in 542 patients (98.2%) and its variant for patients allergic to beta-lactam antibiotics was used in 10 patients (1.8%). The postoperative pharmacotherapeutic protocol assigned depended on whether a central venous line (317 patients; 57.4%) or only peripheral venous access (235 patients; 42.6%) was used. CONCLUSIONS: Each clinical unit has a duty to carry out and update consensus protocols (always based on the best scientific evidence available) that can be used in the processes managed in that unit. The use of these protocols shows a high degree of acceptance among physicians and nurses, allowing clinical practice to be standardized and healthcare processes to be homogenized.

Detección de patrones interactivos en la comunicación de familias con hijos adolescentes / Interactive patterns detection in family communication with adolescents

La comunicación próxima es indicador relevante para valorar la funcionalidad familiar. El objetivo es analizar esta comunicación para describir el tipo de liderazgo, los patrones de interacción y la relación entre elementos verbales y no verbales del discurso. El diseño observacional planteado es un diseño idiográfico, puntual y multidimensional, que utiliza el formato de campo como instrumento de observación. Los participantes han sido siete familias normalizadas formadas por los progenitores y un hijo adolescente. Los resultados, acordes con el tipo de familias analizadas, muestran un estilo de comunicación predominantemente democrático, ejercido por los adultos y con frecuentes expresiones de apoyo. El análisis secuencial incorpora únicamente las categorías del criterio emisor, y detecta secuencias significativas que muestran interacciones simétricas entre los tres miembros. Los canales verbal y no verbal aportan informaciones complementarias. Aparecen patrones diferentes en función del género del hijo/a adolescente

Nondistant communication is a relevant indicator for family functionality valuation. The goal of this study is to analyze this communication in order to identify specific kinds of leadership, interaction patterns and the relation between verbal and nonverbal elements in communication. The observational design exposed is an idiographic one, punctual and multidimensional, which uses field format as observation instrument. Participants were seven standardized families made up of both ancestors and an adolescent son or daughter. According to the family models analyzed, results show a predominantly democratic communication style in adults with recurrent support expressions. The sequential analysis incorporates only categories from the emitter point of view, and detects relevant sequences which show symmetric interaction between all three family members. Verbal and nonverbal channels provide complementary information. Depending on adolescents’ gender different patterns in behaviour can be identified as well

Prescripción electrónica asistida de protocolos farmacoterapéuticos en una unidad de cirugía coloproctológica / Computerized physician order entry of pharmacotherapeutic protocols in a colorectal surgery unit

Introducción. Tras la implantación en nuestro hospital de los sistemas de prescripción electrónica, presentamos los protocolos farmacoterapéuticos que empleamos en la atención de nuestros pacientes. Material y métodos. Diseño por consenso de 6 protocolos farmacoterapéuticos en una unidad de cirugía coloproctológica y aplicación de éstos durante un período de 2 años. Se excluye del estudio a los pacientes intervenidos de urgencias y a los intervenidos en el programa de cirugía sin ingreso. Resultados. Con este recurso se trató a 772 pacientes. El 28,5% se asignó al protocolo de preoperatorio en cirugía proctológica menor. Una vez intervenidos se incluyó a todos ellos en el protocolo de postoperatorio de cirugía proctológica menor. Se asignó a los 552 pacientes restantes a los protocolos de cirugía colorrectal mayor. El protocolo de preoperatorio en cirugía colorrectal mayor se indicó en 542 casos y su variante para alérgicos a los betalactámicos, en 10 casos. El protocolo postoperatorio asignado dependió de si el paciente tenía colocada una vía venosa central (57,4%) o si, por el contrario, sólo disponía de una o varias vías periféricas (42,6%). Conclusiones. Es responsabilidad de cada unidad clínica la realización y actualización, siempre con las mejores pruebas científicas disponibles, de los protocolos consensuados que se pueden aplicar a los procesos atendidos en ella. La utilización de éstos muestra un alto grado de aceptación por parte de los prescriptores y de enfermería, permitiendo estandarizar la práctica clínica y homogeneizar los procesos asistenciales (AU)

Introduction. Computerized physician order entry was introduced in our hospital. We present the pharmacotherapeutic protocols that we use in patient care. Material and methods. Six pharmaceutical protocols were designed by consensus in a colorectal surgery unit and were applied over a 2-year period. Patients undergoing ambulatory or urgent surgery were excluded. Results. We treated 772 patients. Two hundred twenty patients (28.5%) were assigned to the preoperative protocol for minor proctologic surgery. After surgery, all 220 patients were included in the postoperative protocol for minor proctologic surgery. The remaining 552 patients (71.5%) were assigned to the protocol for major colorectal surgery. The preoperative protocol for major colorectal surgery was indicated in 542 patients (98.2%) and its variant for patients allergic to beta-lactam antibiotics was used in 10 patients (1.8%). The postoperative pharmacotherapeutic protocol assigned depended on whether a central venous line (317 patients; 57.4%) or only peripheral venous access (235 patients; 42.6%) was used. Conclusions. Each clinical unit has a duty to carry out and update consensus protocols (always based on the best scientific evidence available) that can be used in the processes managed in that unit. The use of these protocols shows a high degree of acceptance among physicians and nurses, allowing clinical practice to be standardized and healthcare processes to be homogenized (AU)