Transcriptomic profiles and diagnostic biomarkers in the Mediterranean seagrasses Posidonia oceanica and Cymodocea nodosa reveal mechanistic insights of adaptative strategies upon desalination brine stress.

Seawater desalination by reverse osmosis is growing exponentially due to water scarcity. Byproducts of this process (e.g. brines), are generally discharged directly into the coastal ecosystem, causing detrimental effects, on benthic organisms. Understanding the cellular stress response of these organisms (biomarkers), could be crucial for establishing appropriate salinity thresholds for discharged brines. Early stress biomarkers can serve as valuable tools for monitoring the health status of brine-impacted organisms, enabling the prediction of long-term irreversible damage caused by the desalination industry. In this study, we conducted laboratory-controlled experiments to assess cellular and molecular biomarkers against brine exposure in two salinity-sensitive Mediterranean seagrasses: Posidonia oceanica and Cymodocea nodosa. Treatments involved exposure to 39, 41, and 43 psu, for 6 h and 7 days. Results indicated that photosynthetic performance remained unaffected across all treatments. However, under 43 psu, P. oceanica and C. nodosa exhibited lipid oxidative damage, which occurred earlier in P. oceanica. Additionally, P. oceanica displayed an antioxidant response at higher salinities by accumulating phenolic compounds within 6 h and ascorbate within 7 d; whereas for C. nodosa the predominant antioxidant mechanisms were phenolic compounds accumulation and total radical scavenging activity, which was evident after 7 d of brines exposure. Finally, transcriptomic analyses in P. oceanica exposed to 43 psu for 7 days revealed a poor up-regulation of genes associated with brassinosteroid response and abiotic stress response, while a high down-regulation of genes related to primary metabolism was detected. In C. nodosa, up-regulated genes were involved in DNA repair, cell cycle regulation, and reproduction, while down-regulated genes were mainly associated with photosynthesis and ribosome assembly. Overall, these findings suggest that 43 psu is a critical salinity-damage threshold for both seagrasses; and despite the moderate overexpression of several transcripts that could confer salt tolerance, genes involved in essential biological processes were severely downregulated.

Desalination brine effects beyond excess salinity: Unravelling specific stress signaling and tolerance responses in the seagrass Posidonia oceanica.

Desalination has been proposed as a global strategy for tackling freshwater shortage in the climate change era. However, there is a concern regarding the environmental effects of high salinity brines discharged from desalination plants on benthic communities. In this context, seagrasses such as the Mediterranean endemic and ecologically important Posidonia oceanica have shown high vulnerability to elevated salinities. Most ecotoxicological studies regarding desalination effects are based on salinity increments using artificial sea salts, although it has been postulated that certain additives within the industrial process of desalination may exacerbate a negative impact beyond just the increased salinities of the brine. To assess the potential effect of whole effluent brines on P. oceanica, mesocosm experiments were conducted within 10 days, simulating salinity increment with either artificial sea salts or brines from a desalination plant (at 43 psµ, 6 psµ over the natural 37 psµ). Morphometrical (growth and necrosis), photochemical (PSII chlorophyll a fluorometry), metabolic, such as hydrogen peroxide (H2O2), thiobarbituric reactive substances (TBARS) and ascorbate/dehydroascorbate (ASC/DHA), and molecular (expression of key tolerance genes) responses were analyzed in each different treatment. Although with a still positive leaf growth, associated parameters decreased similarly for both artificial sea salt and brine treatments. Photochemical parameters did not show general patterns, although only P. oceanica under brines demonstrated greater energy release through heat (NPQ). Lipid peroxidation and upregulation of genes related to oxidative stress (GR, MnSOD, and FeSOD) or ion exclusion (SOS3 and AKT2/3) were similarly incremented on both hypersalinity treatments. Conversely, the ASC/DHA ratio was significantly lower, and the expression of SOS1, CAT, and STRK1 was increased under brine influence. This study revealed that although metabolic and photochemical differences occurred under both hypersalinity treatments, growth (the last sign of physiological detriment) was similarly compromised, suggesting that the potential effects of desalination are mainly caused by brine-associated salinities and are not particularly related to other industrial additives.

A risk assessment on Zostera chilensis, the last relict of marine angiosperms in the South-East Pacific Ocean, due to the development of the desalination industry in Chile.

Seagrasses, which are considered among the most ecologically valuable and endangered coastal ecosystems, have a narrowly limited distribution in the south-east Pacific, where Zostera chilensis is the only remaining relict. Due to water scarcity, desalination industry has grown in the last decades in the central-north coasts of Chile, which may be relevant to address in terms of potential impacts on benthic communities due to their associated high-salinity brine discharges to subtidal ecosystems. In this work, we assessed ecophysiological and cellular responses to desalination-extrapolable hypersalinity conditions on Z. chilensis. Mesocosms experiments were performed for 10 days, where plants were exposed to 3 different salinity treatments: 34 psu (control), 37 psu and 40 psu. Photosynthetic performance, H2O2 accumulation, and ascorbate content (reduced and oxidized) were measured, as well as relative gene expression of enzymes related to osmotic regulation and oxidative stress; these, at 1, 3, 6 and 10 days. Z. chilensis showed a decrease in photosynthetic parameters such as electron transport rate (ETRmax) and saturation irradiance (EkETR) under hypersalinity treatments, while non-photochemical quenching (NPQmax) presented an initial increment and a subsequent decline at 40 psu. H2O2 levels increased with hypersalinity, while ascorbate and dehydroascorbate only increased under 37 psu, although decreased along the experimental period. Increased salinities also triggered the expression of genes related to ion transport and osmolyte syntheses, but salinity-dependent up-regulated genes were mostly those related to the reactive oxygen species metabolism. The relict seagrass Z. chilensis has shown to withstand increased salinities that may be extrapolable to desalination effects in the short-term. As the latter is not fully clear in the long-term, and considering the restricted distribution and ecological importance, direct brine discharges to Z. chilensis meadows may not be recommended.

Macroalgae metal-biomonitoring in Antarctica: Addressing the consequences of human presence in the white continent.

Marine ecosystems in the Arctic and Antarctica were once thought pristine and away from important human influence. Today, it is known that global processes as atmospheric transport, local activities related with scientific research bases, military and touristic maritime traffic, among others, are a potential source of pollutants. Macroalgae have been recognized as reliable metal-biomonitoring organisms due to their accumulation capacity and physiological responses. Metal accumulation (Al, Cd, Cu, Fe, Pb, Zn, Se, and Hg) and photosynthetic parameters (associated with in vivo chlorophyll a fluorescence) were assessed in 77 samples from 13 different macroalgal species (Phaeophyta; Chlorophyta; Rhodophyta) from areas with high human influence, nearby research and sometimes military bases and a control area, King George Island, Antarctic Peninsula. Most metals in macroalgae followed a pattern influenced by rather algal lineage than site, with green seaweeds displaying trends of higher levels of metals as Al, Cu, Cr and Fe. Photosynthesis was also not affected by site, showing healthy organisms, especially in brown macroalgae, likely due to their great dimensions and morphological complexity. Finally, data did not demonstrate a relationship between metal accumulation and photosynthetic performance, evidencing low anthropogenic-derived impacts associated with metal excess in the area. Green macroalgae, especially Monostroma hariotti, are highlighted as reliable for further metal biomonitoring assessments. In the most ambitious to date seaweed biomonitoring effort conducted towards the Austral pole, this study improved by 91% the overall knowledge on metal accumulation in macroalgae from Antarctica, being the first report in species as Sarcopeltis antarctica and Plocamium cartilagineum. These findings may suggest that human short- and long-range metal influence on Antarctic coastal ecosystems still remains under control.

Ulva compressa from Copper-Polluted Sites Exhibits Intracellular Copper Accumulation, Increased Expression of Metallothioneins and Copper-Containing Nanoparticles in Chloroplasts.

In order to analyze the mechanisms involved in copper accumulation in Ulva compressa, algae were collected at control sites of central and northern Chile, and at two copper-polluted sites of northern Chile. The level of intracellular copper, reduced glutathione (GSH), phytochelatins (PCs), PC2 and PC4, and transcripts encoding metallothioneins (MTs) of U. compressa, UcMT1, UcMT2 and UcMT3, were determined. Algae of control sites contained around 20 µg of copper g-1 of dry tissue (DT) whereas algae of copper-polluted sites contained 260 and 272 µg of copper g-1 of DT. Algae of control sites and copper-polluted sites did not show detectable amounts of GSH, the level of PC2 did not change among sites whereas PC4 was increased in one of the copper-polluted sites. The level of transcripts of UcMT1 and UcMT2 were increased in algae of copper-polluted sites, but the level of UcMT3 did not change. Algae of a control site and a copper-polluted site were visualized by transmission electron microscopy (TEM) and the existence of copper in electrodense particles was analyzed using energy dispersive x-ray spectroscopy (EDXS). Algae of copper-polluted sites showed electrodense nanoparticles containing copper in the chloroplasts, whereas algae of control sites did not. Algae of a control site, Cachagua, were cultivated without copper (control) and with 10 µM copper for 5 days and they were analyzed by TEM-EDXS. Algae cultivated with copper showed copper-containing nanoparticles in the chloroplast whereas control algae did not. Thus, U. compressa from copper-polluted sites exhibits intracellular copper accumulation, an increase in the level of PC4 and expression of UcMTs, and the accumulation of copper-containing particles in chloroplasts.

Becas Salud Investiga a proyectos de Investigación 2020-2021. Estudio exploratorio descriptivo sobre la accesibilidad a la interrupción legal del embarazo en dos áreas programáticas de la provincia del Chubut, en el marco de la Ley XV/14. Estudio en dos etapas, desde la experiencia de personas que accedieron a la práctica y desde la experiencia de los equipos de salud

La presente investigación tuvo el objetivo de caracterizar la accesibilidad existente para la interrupción legal del embarazo (ILE) en el marco de la Ley XV 14 en dos Áreas Programáticas de la provincia del Chubut desde la sanción de la misma hasta la actualidad desde la perspectiva de las personas que accedieron a la práctica y de los equipos de salud. Busca identificar los obstáculos y facilitadores en el acceso a la práctica. Chubut posee una Ley pionera en el país, que se sancionó en el año 2010 y que da marco a la práctica de la ILE. Sin embargo, continúan presentándose diferentes dificultades en el ejercicio del derecho allí consagrado. Se realizó una investigación cualitativa a través del diseño de un estudio exploratorio descriptivo, transversal y retrospectivo dado que se trató de una primera aproximación al tema en Chubut y era necesario conocer la situación en profundidad. Primero, se trabajó con una muestra de la población de personas que hayan solicitado la ILE en ambas Áreas Programáticas buscando estudiar la accesibilidad desde una concepción vincular. Luego, se buscó conocer la experiencia de los equipos de salud. Se realizaron entrevistas a partir de una guía semi estructurada elaborada en función de los objetivos y principales categorías de análisis. Se realizó un análisis temático de la información. En las Áreas Programáticas estudiadas se objetivaron diferentes obstáculos en el acceso a la ILE en sus dimensiones geográfica, económica, organizacional, simbólica e informativa. Se identificaron facilitadores que se relacionan, principalmente, con la existencia de redes informales de cuidado que se ponen en juego durante el proceso. Incorporar el estudio de la accesibilidad desde una perspectiva vincular permite incluir la mirada de diferentes actores sociales que se vinculan para el acceso al derecho a la ILE en el territorio y da lugar a su comprensión como proceso de salud-enfermedad-cuidado inmerso en redes formales e informales de cuidado.

Antarctic intertidal macroalgae under predicted increased temperatures mediated by global climate change: Would they cope?

The Antarctic Peninsula is one of the regions to be most affected by increase in sea surface temperatures (SSTs) mediated by Global Climate Change; indeed, most negative predictions imply an up to 6 °C increment by the end of the XXI century. Temperature is one of the most important factors mediating diversity and distribution of macroalgae, although there is still no consensus as to the likely effects of higher SSTs, especially for polar seaweeds. Some available information suggests that potential strategies to withstand future increases in SSTs will be founded upon the glutathione-ascorbate cycle and the induction of chaperone-functioning heat shock proteins (HSPs); however, their eventual role, even for general stress responses, is unclear. The intertidal green, brown and red macroalgae species Monostroma hariotii, Adenocystis utricularis and Pyropia endiviifolia, respectively, from King George Island, Antarctic Peninsula, were exposed to 2 °C (control) and 8 °C (climate change scenario) for up to 5 days (d). Photosynthetic activity (αETR and ETRmax, and EkETR), photoinhibition (Fv/Fm) and photoprotection processes (αNPQ, NPQmax, and EkNPQ) provided no evidence of negative ecophysiological effects. There were moderate increases in H2O2 production and levels of lipid peroxidation with temperature, results supported by stable levels of total glutathione and ascorbate pools, with mostly higher levels of reduced ascorbate and glutathione than oxidized forms in all species. Transcripts of P. endiviifolia indicated a general upregulation of all antioxidant enzymes and HSPs genes studied under warmer temperature, although with different levels of activation with time. This pioneering investigation exploring different levels of biological organization, suggested that Antarctic intertidal macroalgae may be able to withstand future rise in SSTs, probably slightly altering their latitudinal distribution and/or range of thermal tolerance, by exhibiting robust glutathione-ascorbate production and recycling, as well as the induction of associated antioxidant enzymatic machinery and the syntheses of HSPs.

Physiological and metabolic responses to hypersalinity reveal interpopulation tolerance in the green macroalga Ulva compressa with different pollution histories.

There is scarce investigation addressing interpopulation tolerance responses to address the influence of a history of chronic stress exposure, as that occurring in polluted environments, in photoautotrophs. We evaluated ecophysiological (photosynthetic activity) and metabolic (oxidative stress and damage) responses of two populations of green macroalga Ulva compressa from polluted (Ventanas) and non-polluted (Cachagua) localions of central Chile, and exposed to controlled hypersalinity conditions of 32 (control), 42, 62 and 82 psu (practical salinity units) for 6 h, 48 h and 6 d. Both primary production (ETRmax) and photosynthetic efficiency (αETR) were generally higher in the population from Cachagua compared to Ventanas at all times and salinities. Moreover, at most experimental times and salinities the population from Ventanas had greater levels of H2O2 and lipid peroxidation that individuals from Cachagua. Total ascorbate was higher in the population of Cachagua than Ventanas at 42 and 82 psu after 6 and 48 h, respectively, while at 6 d concentrations were similar between both populations at all salinities. Total glutathione was greater in both populations after 6 h at all salinities, but at 48 h its concentrations were higher only in the population from Cachagua, a trend that was maintained at 6 d under 82 psu only. Reduced and oxidized ascorbate (ASC and DHA, respectively) and glutathione (GSH and GSSG, respectively) demonstrated similar patterns between U. compressa populations, with an increase oxidation with greater salinities but efficient recycling to maintain sufficient batch of ASC and GSH. When assessing the expression of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR), while the population of Ventanas displayed a general trend of upregulation with increasing salinities along the experiments, U. compressa from Cachagua revealed patterns of downregulation. Results demonstrated that although both populations were still viable after the applied hypersalinities during all experimental times, biological performance was usually more affected in the population from the Ventanas than Cachagua, likely due to a depressed baseline metabolism after a long history of exposition to environmental pollution.

Assessment of the anti-inflammatory and engraftment potential of horse endometrial and adipose mesenchymal stem cells in an in vivo model of post breeding induced endometritis.

Horse mesenchymal stem cells (MSC) are potential anti-inflammatory tools for post-breeding induced endometritis (PBIE). In this research MSCs isolated from the endometrium or subcutaneous fat of the same donors were infused iu into mares with PBIE for assessment of their anti-inflammatory action and engraftment. PBIE was induced in nine gynecologically healthy mares by iu infusion of 500 million dead sperm in saline. Inflammatory markers were analyzed in uterine lavages and biopsies immediately before (phase I) and 3 h after infusion of sperm (phase II). Measurements: polymorph nuclear cells (PMN), proteins IL-6 and TNFα (ELISA in the lavages) and immunostaining in biopsies, transcripts of IL-1α, 6, 8, 10, TNFα and COX2 (qPCR of pelleted lavages). At 24 h after sperm deposition (phase III), mares were instilled iu with 20 ml of saline containing 2 × 107 adipose MSCs (n = 3, group 1) or endometrial MSCs (n = 3, group 2). Cells were labeled previously with carboxyfluorescein diacetate succinimidyl ester (CFDA SE). A third group (n = 3) received 20 mL of sterile saline alone. After 48 h another biopsy/lavage were done and the same parameters analyzed. For engraftment, additional biopsies were taken at days 10 and 30 of sperm infusion and analyzed by confocal microscopy. Dead sperm in saline markedly increased PMNs counts, IL-6 and TNFα expression in the ELISA (p < 0.05) and immunostaining. In phase III a significant reduction (p < 0.0001) of PMN was found in all samples, including control mares. A decrease (p < 0.05) of IL-6 and TNF-α was detected by ELISA, in the groups that received MSC, but not in control group. In the aMSC-treated group, a significant decrease was found in the expression of (IL1α, p = 0.0003; IL-6 p 0.04; IL-8, p = 0.006, TNFα p = 0.004). Expression of IL-10 and COX2 remained unchanged (p = 0.08). In the mares that received eMSC, IL-6 and 8 decreased significantly (p = 0.01), IL-10 increased (p = 0.009), while TNFα, COX2 and IL1α did not significantly change their expression. In the engraftment experiment CFDA label was found sparingly in all the samples analyzed until day 30, mainly at the stromal compartment of the endometrium. No differences in the engraftment pattern was found among cell origins. We conclude that inoculation of MSCs significantly reduced inflammation independently of the origin of the cells and that cells perform limited engraftment detectable after one month of infusion. These findings can be of help for the design of new anti-inflammatory therapies of uterine diseases in mares.

[Assessment of two frailty scales for the preoperative period]. / Fragilidad: en busca de herramientas de evaluación preoperatoria.

BACKGROUND: In the perioperative context, a frailty evaluation scale must consider certain characteristics such as validation, execution speed, simplicity, the capacity to measure multiple dimensions and not being dependent on a cognitive or physical test that could not be performed prior to surgery. The test should select patients that could benefit from interventions aimed to improve their postoperative outcomes. AIM: To validate two frailty evaluation scales for the perioperative period. MATERIAL AND METHODS: The Risk Analysis Index with local modifications (RAI-M) were applied to 201 patients aged 73 ± 7 years (49% women) and the Edmonton frailty scale were applied in 151 patients aged 73 ± 7 years (49% women) in the preoperative period. Their results were compared with the Rockwood frailty index. RESULTS: The Edmonton frail scale showed adequate psychometric properties and assessed multiple dimensions through 8 of the 11 original questions, achieving a discrimination power over 80% compared to the Rockwood Index. The RAI- M, demonstrated solid psychometric properties with a tool that examines 4 dimensions of frailty through 15 questions and reviewing the presence of 11 medical comorbidities. This scale had a discrimination power greater than 85% and it was significantly associated with prolongation of the planned hospital stay and mortality. CONCLUSIONS: RAI-M is a short and easily administered scale, useful to detect frailty in the preoperative period.

Fragilidad: en busca de herramientas de evaluación preoperatoria / Assessment of two frailty scales for the preoperative period

Background: In the perioperative context, a frailty evaluation scale must consider certain characteristics such as validation, execution speed, simplicity, the capacity to measure multiple dimensions and not being dependent on a cognitive or physical test that could not be performed prior to surgery. The test should select patients that could benefit from interventions aimed to improve their postoperative outcomes. Aim: To validate two frailty evaluation scales for the perioperative period. Material and Methods: The Risk Analysis Index with local modifications (RAI-M) were applied to 201 patients aged 73 ± 7 years (49% women) and the Edmonton frailty scale were applied in 151 patients aged 73 ± 7 years (49% women) in the preoperative period. Their results were compared with the Rockwood frailty index. Results: The Edmonton frail scale showed adequate psychometric properties and assessed multiple dimensions through 8 of the 11 original questions, achieving a discrimination power over 80% compared to the Rockwood Index. The RAI- M, demonstrated solid psychometric properties with a tool that examines 4 dimensions of frailty through 15 questions and reviewing the presence of 11 medical comorbidities. This scale had a discrimination power greater than 85% and it was significantly associated with prolongation of the planned hospital stay and mortality. Conclusions: RAI-M is a short and easily administered scale, useful to detect frailty in the preoperative period.

Accesibilidad a la Interrupción Legal del Embarazo en Chubut desde las experiencias de personas que accedieron a la práctica y de los equipos de salud / Accessibility to the Legal Interruption of Pregnancy in Chubut from the experiences of people who accessed the practice and health teams

La presentee investigación tuvo el objetivo de caracterizar la accesibilidad existente para la interrupción legal del embarazo (ILE) en el marco de la ley XV 14 en dos Áreas Programáticas de la provincia del Chubut desde la sanción de la misma hasta la actualidad desde la perspectiva de las personas que accedieron a la práctica y de los equipos de salud. Busca identificar los obstáculos y facilitadores en el acceso a la práctica. Chubut posee una Ley pionera en el país, que se sancionó en el año 2010 y que da marco a la práctica de la ILE. Sin embargo, continúan presentándosee diferentes dificultades en el ejercicio del derecho allí consagrado. Se realizó una investigación cualitativa a través del diseño de un estudio exploratorio descriptivo, transversal y retrospectivo dado que se trató de una primera aproximación al tem en Chubut y era necesario conocer la situación en profundidad. Primero, se trabajó con una muestra de la población de personas que hayan solicitado la ILE en ambas Áreas Programáticas buscando estudiar la accesibilidad desde una concepción vincular. Luego, se buscó conocer la experiencia de los equipos de salud. Se realizaron entrevistas a partir de una guía semi estructurada elaborada en función de los objetivos y principales categorías de análisis. Se realizó un análisis temático de la información. En las Áreas Programáticas estudiadas se objetivaron diferentes obstáculos en el acceso a la ILE en sus dimensiones geográfica, económicca, organizacional, simbólica e informativa. Se identificaron facilitadores que se relacionan, principalmente, con la existencia de redes informales de cuidado que se ponen en juego durante el proceso. Incorporar el estudio de la accesibilidad desde una perspectiva vincular permite incluir la mirada de diferentes actores sociales que se vinculan para el acceso al derecho a la ILE en el territorio y da lugar a su comprensión como proceso de salud-enfermedad-cuidado inmerso en redes formales e informales de cuidado

MAPK Pathway under Chronic Copper Excess in Green Macroalgae (Chlorophyta): Involvement in the Regulation of Detoxification Mechanisms.

Following the physiological complementary/parallel Celis-Plá et al., by inhibiting extracellular signal regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and cytokinin specific binding protein (p38), we assessed the role of the mitogen-activated protein kinases (MAPK) pathway in detoxification responses mediated by chronic copper (10 µM) in U. compressa. Parameters were taken at 6, 24, and 48 h, and 6 days (d). H2O2 and lipid peroxidation under copper and inhibition of ERK, JNK, or p38 alone increased but recovered by the sixth day. By blocking two or more MAPKs under copper, H2O2 and lipid peroxidation decayed even below controls. Inhibition of more than one MAPK (at 6 d) caused a decrease in total glutathione (reduced glutathione (GSH) + oxidised glutathione (GSSG)) and ascorbate (reduced ascorbate (ASC) + dehydroascorbate (DHA)), although in the latter it did not occur when the whole MAPK was blocked. Catalase (CAT), superoxide dismutase (SOD), thioredoxin (TRX) ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), and glutathione synthase (GS), were downregulated when blocking more than one MAPK pathway. When one MAPK pathway was blocked under copper, a recovery and even enhancement of detoxification mechanisms was observed, likely due to crosstalk within the MAPKs and/or other signalling processes. In contrast, when more than one MAPK pathway were blocked under copper, impairment of detoxification defences occurred, demonstrating that MAPKs were key signalling mechanisms for detoxification in macroalgae.

MAPK Pathway under Chronic Copper Excess in Green Macroalgae (Chlorophyta): Influence on Metal Exclusion/Extrusion Mechanisms and Photosynthesis.

There is currently no information regarding the role that whole mitogen activated protein kinase (MAPK) pathways play in counteracting environmental stress in photosynthetic organisms. To address this gap, we exposed Ulva compressa to chronic levels of copper (10 µM) specific inhibitors of Extracellular Signal Regulated Kinases (ERK), c-Jun N-terminal Kinases (JNK), and Cytokinin Specific Binding Protein (p38) MAPKs alone or in combination. Intracellular copper accumulation and photosynthetic activity (in vivo chlorophyll a fluorescence) were measured after 6 h, 24 h, 48 h, and 6 days of exposure. By day 6, when one (except JNK) or more of the MAPK pathways were inhibited under copper stress, there was a decrease in copper accumulation compared with algae exposed to copper alone. When at least two MAPKs were blocked, there was a decrease in photosynthetic activity expressed in lower productivity (ETRmax), efficiency (αETR), and saturation of irradiance (EkETR), accompanied by higher non-photochemical quenching (NPQmax), compared to both the control and copper-only treatments. In terms of accumulation, once the MAPK pathways were partially or completely blocked under copper, there was crosstalk between these and other signaling mechanisms to enhance metal extrusion/exclusion from cells. Crosstalk occurred among MAPK pathways to maintain photosynthesis homeostasis, demonstrating the importance of the signaling pathways for physiological performance. This study is complemented by a parallel/complementary article Rodríguez-Rojas et al. on the role of MAPKs in copper-detoxification.

Cadmium and/or copper excess induce interdependent metal accumulation, DNA methylation, induction of metal chelators and antioxidant defences in the seagrass Zostera marina.

In this investigation, we assessed the effects of Cu and/or Cd excess on physiological and metabolic processes of the widespread seagrass Zostera marina. Adult were exposed to low Cd and Cu (0.89 and 0.8 µM, respectively) and high Cd and Cu (8.9 and 2.4 µM, respectively) for 6 d at: Control conditions; low Cu; high Cu; low Cd; high Cd; low Cd and low Cu; and high Cd and high Cu. Photosynthetic performance decreased under single and combined treatments, although effects were more negative under Cu than Cd. Total Cu accumulation was higher than Cd, under single and combined treatments; however, their accumulation was generally lower when applied together, suggesting competition among them. Levels of glutathione (GSH) and phytochelatins (PCs) followed patterns similar to metal accumulation, with up to PC5, displaying adaptations in tolerance. A metallothionein (MET) gene showed upregulation only at high Cd, low Cu, and high Cu. The expression of the enzymes glutathione reductase (GR), ascorbate peroxidase (APX), and catalase (CAT) was greatest at high Cu, and at high Cd and Cu together; the highest expression was under Cu, alone and combined. Both metals induced upregulation of the DNA methyltransferases CMT3 and DRM2, with the highest expression at single Cu. The DNA demethylation ROS1 was overexpressed in treatments containing high Cu, suggesting epigenetic modifications. The results show that under copper and/or cadmium, Z. marina was still biologically viable; certainly based, at least in part, on the induction of metal chelators, antioxidant defences and methylation/demethylation pathways of gene regulation.

Diferencias en temperamento según nivel socioeconómico en niños chilenos de 12 y 30 meses / Differences in temperament according to socioeconomic status in 12 to 30 months Chilean children

Resumen Se entiende el temperamento como el conjunto de diferencias individuales en reactividad y autorregulación, manteniéndose relativamente estable a través del tiempo, y siendo influenciado por la herencia, la maduración biológica y el contacto con el ambiente. En el presente estudio se buscó establecer si se dan diferencias en las dimensiones de temperamento de extraversión, afecto negativo y autorregulación según nivel socioeconómico, en niños chilenos asistentes a sala cuna. Se evaluaron 103 niños de ambos sexos, a los 12 y a los 30 meses de edad, con los cuestionarios IBQ-R-VSF y ECBQ-VSF, los cuales respondieron los padres. Los resultados mostraron que entre los 12 y 30 meses la extraversión se mantuvo constante, mientras que las dimensiones de afecto negativo y autorregulación disminuyeron de manera significativa. A los 12 meses los niños de NSE alto mostraron una mayor autorregulación, mientras que a los 30 meses los niños de NSE bajo evidenciaron un mayor afecto negativo. Se discuten los resultados en base a la evidencia encontrada sobre las diferentes realidades en los distintos niveles socioeconómicos.

Abstract Temperament is understood as the set of individual differences in reactivity and self-regulation, considered as stable through time, and influenced by the heritability, the biological maturation, and the interaction with the environment. The aim of this study was to determine if the dimensions of temperament (extraversion, negative affect, and self-regulation) show differences according to socioeconomic status in Chilean children attending to nurseries. For this study, 103 children from both genders were evaluated at 12 and 30 months old, using the IBQ-R-VSF and ECBQ-VSF questionnaires, which were answered by their parent. Results showed that between 12 and 30 months, extraversion remained constant, while the dimensions of negative affect and self-regulation decreased significantly. Children from high socioeconomic status showed higher self-regulation at age of 12 months, while the ones from low socioeconomic status decreased their negative affect at 30 months old. Results are discussed based on evidence about realities from different socioeconomic statuses.

Role of GAD peptides p217 and p290 in the repair of INS receptor in salivary tissues of type 1 diabetic mice.

Glutamate decarboxylase or glutamic acid decarboxylase (GAD) is a protein associated with autoimmune diseases, including type-1 diabetes. This disease is primarily associated with the occurrence of a specific isoform: GAD65. Conversely, some specific peptides of this protein may block autoimmunity in diabetes. In this respect, understanding the relationship between GAD and the development of diabetes is important, and it is necessary to understand the role of each GAD peptide to design effective autoimmune diabetes treatments. The purpose of the present study was to analyze the effects of treatment with GAD-derived peptides p217 and p290 on INS receptors in the salivary epithelium of nonobese diabetic (NOD) animals. Three groups of 7 mice each were studied: I, BALB/c mice (control); II, NOD mice; and III, NOD mice treated with peptides p290 and p217. Groups I and II only received buffered saline solution. Glucose levels were measured daily during the 21 days of the experiment. After the study, the animals were euthanized and the parotid and submandibular glands were removed for the analysis of INS-R by fluorescence microscopy. Therapy with two peptides together was associated with reduced glucose levels in NOD mice and intense INS-R expression in both salivary organs. Our approach of combining GAD p217 and p290 peptides contributed to hormonal balance and promoted the repair of INS-R.

Flavoprotein-Mediated Tellurite Reduction: Structural Basis and Applications to the Synthesis of Tellurium-Containing Nanostructures.

The tellurium oxyanion tellurite (TeO3 (2-)) is extremely harmful for most organisms. It has been suggested that a potential bacterial tellurite resistance mechanism would consist of an enzymatic, NAD(P)H-dependent, reduction to the less toxic form elemental tellurium (Te(0)). To date, a number of enzymes such as catalase, type II NADH dehydrogenase and terminal oxidases from the electron transport chain, nitrate reductases, and dihydrolipoamide dehydrogenase (E3), among others, have been shown to display tellurite-reducing activity. This activity is generically referred to as tellurite reductase (TR). Bioinformatic data resting on some of the abovementioned enzymes enabled the identification of common structures involved in tellurite reduction including vicinal catalytic cysteine residues and the FAD/NAD(P)(+)-binding domain, which is characteristic of some flavoproteins. Along this line, thioredoxin reductase (TrxB), alkyl hydroperoxide reductase (AhpF), glutathione reductase (GorA), mercuric reductase (MerA), NADH: flavorubredoxin reductase (NorW), dihydrolipoamide dehydrogenase, and the putative oxidoreductase YkgC from Escherichia coli or environmental bacteria were purified and assessed for TR activity. All of them displayed in vitro TR activity at the expense of NADH or NADPH oxidation. In general, optimal reducing conditions occurred around pH 9-10 and 37°C. Enzymes exhibiting strong TR activity produced Te-containing nanostructures (TeNS). While GorA and AhpF generated TeNS of 75 nm average diameter, E3 and YkgC produced larger structures (>100 nm). Electron-dense structures were observed in cells over-expressing genes encoding TrxB, GorA, and YkgC.

Characterization of cytokinin signaling and homeostasis gene families in two hardwood tree species: Populus trichocarpa and Prunus persica.

BACKGROUND: Through the diversity of cytokinin regulated processes, this phytohormone has a profound impact on plant growth and development. Cytokinin signaling is involved in the control of apical and lateral meristem activity, branching pattern of the shoot, and leaf senescence. These processes influence several traits, including the stem diameter, shoot architecture, and perennial life cycle, which define the development of woody plants. To facilitate research about the role of cytokinin in regulation of woody plant development, we have identified genes associated with cytokinin signaling and homeostasis pathways from two hardwood tree species. RESULTS: Taking advantage of the sequenced black cottonwood (Populus trichocarpa) and peach (Prunus persica) genomes, we have compiled a comprehensive list of genes involved in these pathways. We identified genes belonging to the six families of cytokinin oxidases (CKXs), isopentenyl transferases (IPTs), LONELY GUY genes (LOGs), two-component receptors, histidine containing phosphotransmitters (HPts), and response regulators (RRs). All together 85 Populus and 45 Prunus genes were identified, and compared to their Arabidopsis orthologs through phylogenetic analyses. CONCLUSIONS: In general, when compared to Arabidopsis, differences in gene family structure were often seen in only one of the two tree species. However, one class of genes associated with cytokinin signal transduction, the CKI1-like family of two-component histidine kinases, was larger in both Populus and Prunus than in Arabidopsis.