2-Cys peroxiredoxins contribute to thylakoid lipid unsaturation by affecting ω-3 fatty acid desaturase 8.

Fatty acid unsaturation levels affect chloroplast function and plant acclimation to environmental cues. However, the regulatory mechanism(s) controlling fatty acid unsaturation in thylakoid lipids is poorly understood. Here, we have investigated the connection between chloroplast redox homeostasis and lipid metabolism by focusing on 2-Cys peroxiredoxins (Prxs), which play a central role in balancing the redox state within the organelle. The chloroplast redox network relies on NADPH-dependent thioredoxin reductase C (NTRC), which controls the redox balance of 2-Cys Prxs to maintain the reductive activity of redox-regulated enzymes. Our results show that Arabidopsis (Arabidopsis thaliana) mutants deficient in 2-Cys Prxs contain decreased levels of trienoic fatty acids, mainly in chloroplast lipids, indicating that these enzymes contribute to thylakoid membrane lipids unsaturation. This function of 2-Cys Prxs is independent of NTRC, the main reductant of these enzymes, hence 2-Cys Prxs operates beyond the classic chloroplast regulatory redox system. Moreover, the effect of 2-Cys Prxs on lipid metabolism is primarily exerted through the prokaryotic pathway of glycerolipid biosynthesis and fatty acid desaturase 8 (FAD8). While 2-Cys Prxs and FAD8 interact in leaf membranes as components of a large protein complex, the levels of FAD8 were markedly decreased when FAD8 is overexpressed in 2-Cys Prxs-deficient mutant backgrounds. These findings reveal a function for 2-Cys Prxs, possibly acting as a scaffold protein, affecting the unsaturation degree of chloroplast membranes.

Redox regulation of chloroplast metabolism.

Regulation of enzyme activity based on thiol-disulfide exchange is a regulatory mechanism in which the protein disulfide reductase activity of thioredoxins (TRXs) plays a central role. Plant chloroplasts are equipped with a complex set of up to 20 TRXs and TRX-like proteins, the activity of which is supported by reducing power provided by photosynthetically reduced ferredoxin (FDX) with the participation of a FDX-dependent TRX reductase (FTR). Therefore, the FDX-FTR-TRXs pathway allows the regulation of redox-sensitive chloroplast enzymes in response to light. In addition, chloroplasts contain an NADPH-dependent redox system, termed NTRC, which allows the use of NADPH in the redox network of these organelles. Genetic approaches using mutants of Arabidopsis (Arabidopsis thaliana) in combination with biochemical and physiological studies have shown that both redox systems, NTRC and FDX-FTR-TRXs, participate in fine-tuning chloroplast performance in response to changes in light intensity. Moreover, these studies revealed the participation of 2-Cys peroxiredoxin (2-Cys PRX), a thiol-dependent peroxidase, in the control of the reducing activity of chloroplast TRXs as well as in the rapid oxidation of stromal enzymes upon darkness. In this review, we provide an update on recent findings regarding the redox regulatory network of plant chloroplasts, focusing on the functional relationship of 2-Cys PRXs with NTRC and the FDX-FTR-TRXs redox systems for fine-tuning chloroplast performance in response to changes in light intensity and darkness. Finally, we consider redox regulation as an additional layer of control of the signaling function of the chloroplast.

Characterization of CYCLOPHILLIN38 shows that a photosynthesis-derived systemic signal controls lateral root emergence.

Photosynthesis in leaves generates fixed-carbon resources and essential metabolites that support sink tissues, such as roots. Two of these metabolites, sucrose and auxin, promote growth in root systems, but the explicit connection between photosynthetic activity and control of root architecture has not been explored. Through a mutant screen to identify pathways regulating root system architecture, we identified a mutation in the Arabidopsis thaliana CYCLOPHILIN 38 (CYP38) gene, which causes accumulation of pre-emergent stage lateral roots. CYP38 was previously reported to stabilize photosystem II (PSII) in chloroplasts. CYP38 expression is enriched in shoots, and grafting experiments show that the gene acts non-cell-autonomously to promote lateral root emergence. Growth of wild-type plants under low-light conditions phenocopies the cyp38 lateral root emergence defect, as does the inhibition of PSII-dependent electron transport or Nicotinamide adenine dinucleotide phosphate (NADPH) production. Importantly, these perturbations to photosynthetic activity rapidly suppress lateral root emergence, which is separate from their effects on shoot size. Supplementary exogenous sucrose largely rescued primary root (PR) growth in cyp38, but not lateral root growth. Auxin (indole-3-acetic acid (IAA)) biosynthesis from tryptophan is dependent on reductant generated during photosynthesis. Consistently, we found that wild-type seedlings grown under low light and cyp38 mutants have highly diminished levels of IAA in root tissues. IAA treatment rescued the cyp38 lateral root defect, revealing that photosynthesis promotes lateral root emergence partly through IAA biosynthesis. These data directly confirm the importance of CYP38-dependent photosynthetic activity in supporting root growth, and define the specific contributions of two metabolites in refining root architecture under light-limited conditions.

A chloroplast redox relay adapts plastid metabolism to light and affects cytosolic protein quality control.

In chloroplasts, thiol-dependent redox regulation is linked to light since the disulfide reductase activity of thioredoxins (Trxs) relies on photo-reduced ferredoxin (Fdx). Furthermore, chloroplasts harbor an NADPH-dependent Trx reductase (NTR) with a joint Trx domain, termed NTRC. The activity of these two redox systems is integrated by the redox balance of 2-Cys peroxiredoxin (Prx), which is controlled by NTRC. However, NTRC was proposed to participate in redox regulation of additional targets, prompting inquiry into whether the function of NTRC depends on its capacity to maintain the redox balance of 2-Cys Prxs or by direct redox interaction with chloroplast enzymes. To answer this, we studied the functional relationship of NTRC and 2-Cys Prxs by a comparative analysis of the triple Arabidopsis (Arabidopsis thaliana) mutant, ntrc-2cpab, which lacks NTRC and 2-Cys Prxs, and the double mutant 2cpab, which lacks 2-Cys Prxs. These mutants exhibit almost indistinguishable phenotypes: in growth rate, photosynthesis performance, and redox regulation of chloroplast enzymes in response to light and darkness. These results suggest that the most relevant function of NTRC is in controlling the redox balance of 2-Cys Prxs. A comparative transcriptomics analysis confirmed the phenotypic similarity of the two mutants and suggested that the NTRC-2-Cys Prxs system participates in cytosolic protein quality control. We propose that NTRC and 2-Cys Prxs constitute a redox relay, exclusive to photosynthetic organisms that fine-tunes the redox state of chloroplast enzymes in response to light and affects transduction pathways towards the cytosol.

NTRC-dependent redox balance of 2-Cys peroxiredoxins is needed for optimal function of the photosynthetic apparatus.

Thiol-dependent redox regulation allows the rapid adaptation of chloroplast function to unpredictable changes in light intensity. Traditionally, it has been considered that chloroplast redox regulation relies on photosynthetically reduced ferredoxin (Fd), thioredoxins (Trxs), and an Fd-dependent Trx reductase (FTR), the Fd-FTR-Trxs system, which links redox regulation to light. More recently, a plastid-localized NADPH-dependent Trx reductase (NTR) with a joint Trx domain, termed NTRC, was identified. NTRC efficiently reduces 2-Cys peroxiredoxins (Prxs), thus having antioxidant function, but also participates in redox regulation of metabolic pathways previously established to be regulated by Trxs. Thus, the NTRC, 2-Cys Prxs, and Fd-FTR-Trxs redox systems may act concertedly, but the nature of the relationship between them is unknown. Here we show that decreased levels of 2-Cys Prxs suppress the phenotype of the Arabidopsis thaliana ntrc KO mutant. The excess of oxidized 2-Cys Prxs in NTRC-deficient plants drains reducing power from chloroplast Trxs, which results in low efficiency of light energy utilization and impaired redox regulation of Calvin-Benson cycle enzymes. Moreover, the dramatic phenotype of the ntrc-trxf1f2 triple mutant, lacking NTRC and f-type Trxs, was also suppressed by decreased 2-Cys Prxs contents, as the ntrc-trxf1f2-Δ2cp mutant partially recovered the efficiency of light energy utilization and exhibited WT rate of CO2 fixation and growth phenotype. The suppressor phenotype was not caused by compensatory effects of additional chloroplast antioxidant systems. It is proposed that the Fd-FTR-Trx and NTRC redox systems are linked by the redox balance of 2-Cys Prxs, which is crucial for chloroplast function.

NADPH Thioredoxin Reductase C and Thioredoxins Act Concertedly in Seedling Development.

Thiol-dependent redox regulation of enzyme activity plays a central role in the rapid acclimation of chloroplast metabolism to ever-fluctuating light availability. This regulatory mechanism relies on ferredoxin reduced by the photosynthetic electron transport chain, which fuels reducing power to thioredoxins (Trxs) via a ferredoxin-dependent Trx reductase. In addition, chloroplasts harbor an NADPH-dependent Trx reductase, which has a joint Trx domain at the carboxyl terminus, termed NTRC. Thus, a relevant issue concerning chloroplast function is to establish the relationship between these two redox systems and its impact on plant development. To address this issue, we generated Arabidopsis (Arabidopsis thaliana) mutants combining the deficiency of NTRC with those of Trxs f, which participate in metabolic redox regulation, and that of Trx x, which has antioxidant function. The ntrc-trxf1f2 and, to a lower extent, ntrc-trxx mutants showed severe growth-retarded phenotypes, decreased photosynthesis performance, and almost abolished light-dependent reduction of fructose-1,6-bisphosphatase. Moreover, the combined deficiency of both redox systems provokes aberrant chloroplast ultrastructure. Remarkably, both the ntrc-trxf1f2 and ntrc-trxx mutants showed high mortality at the seedling stage, which was overcome by the addition of an exogenous carbon source. Based on these results, we propose that NTRC plays a pivotal role in chloroplast redox regulation, being necessary for the activity of diverse Trxs with unrelated functions. The interaction between the two thiol redox systems is indispensable to sustain photosynthesis performed by cotyledons chloroplasts, which is essential for early plant development.

[Breastfeeding and its influence into the cognitive process of Spanish school-children (6 years old), measured by the Wechsler Intelligence Scale]. / Lactancia materna y su influencia en los procesos cognitivos de escolares españoles (6 años de edad), valorada con la escala de inteligencia Wechsler.

Some scientific evidence support that a better cognitive development during the school age is related with breastfeeding. In this study, the potential benefit of breastfeeding duration is evaluated, related to Verbal Comprehension, Perceptual Reasoning, Working Memory and Processing Speed. A total of 103 children, first year of Primary School, six-year-old, (47 boys and 56 girls), were included from different schools in the province of Granada (Spain) at urban, semi-urban and rural areas. The global cognitive capability, as well as some specific intelligence domains which permit a more precise and deeper analysis of the cognitive processes, was evaluated through the Wechsler Intelligence Scale for Children--version IV. The results prove an association, statistically signnificative, between the best values of IQ and the other four WISC-IV indexes and a longer breastfeeding. There is a highly significant (p = 0.000) association between the best scores and those children who were breastfed during 6 months, which validates our hypothesis. The advice of breastfeeding during at least the first six months of life should be reinforced to reduce learning difficulties.

Plastid 2-Cys peroxiredoxins are essential for embryogenesis in Arabidopsis.

The redox couple formed by NADPH-dependent thioredoxin reductase C (NTRC) and 2-Cys peroxiredoxins (Prxs) allows fine-tuning chloroplast performance in response to light intensity changes. Accordingly, the Arabidopsis 2cpab mutant lacking 2-Cys Prxs shows growth inhibition and sensitivity to light stress. However, this mutant also shows defective post-germinative growth, suggesting a relevant role of plastid redox systems in seed development, which is so far unknown. To address this issue, we first analyzed the pattern of expression of NTRC and 2-Cys Prxs in developing seeds. Transgenic lines expressing GFP fusions of these proteins showed their expression in developing embryos, which was low at the globular stage and increased at heart and torpedo stages, coincident with embryo chloroplast differentiation, and confirmed the plastid localization of these enzymes. The 2cpab mutant produced white and abortive seeds, which contained lower and altered composition of fatty acids, thus showing the relevance of 2-Cys Prxs in embryogenesis. Most embryos of white and abortive seeds of the 2cpab mutant were arrested at heart and torpedo stages of embryogenesis suggesting an essential function of 2-Cys Prxs in embryo chloroplast differentiation. This phenotype was not recovered by a mutant version of 2-Cys Prx A replacing the peroxidatic Cys by Ser. Neither the lack nor the overexpression of NTRC had any effect on seed development indicating that the function of 2-Cys Prxs at these early stages of development is independent of NTRC, in clear contrast with the operation of these regulatory redox systems in leaves chloroplasts.

The plastid-localized pfkB-type carbohydrate kinases FRUCTOKINASE-LIKE 1 and 2 are essential for growth and development of Arabidopsis thaliana.

BACKGROUND: Transcription of plastid-encoded genes requires two different DNA-dependent RNA polymerases, a nuclear-encoded polymerase (NEP) and plastid-encoded polymerase (PEP). Recent studies identified two related pfkB-type carbohydrate kinases, named FRUCTOKINASE-LIKE PROTEIN (FLN1 and FLN2), as components of the thylakoid bound PEP complex in both Arabidopsis thaliana and Sinapis alba (mustard). Additional work demonstrated that RNAi-mediated reduction in FLN expression specifically diminished transcription of PEP-dependent genes. RESULTS: Here, we report the characterization of Arabidopsis FLN knockout alleles to examine the contribution of each gene in plant growth, chloroplast development, and in mediating PEP-dependent transcription. We show that fln plants have severe phenotypes with fln1 resulting in an albino phenotype that is seedling lethal without a source of exogenous carbon. In contrast, fln2 plants display chlorosis prior to leaf expansion, but exhibit slow greening, remain autotrophic, can grow to maturity, and set viable seed. fln1 fln2 double mutant analysis reveals haplo-insufficiency, and fln1 fln2 plants have a similar, but more severe phenotype than either single mutant. Normal plastid development in both light and dark requires the FLNs, but surprisingly skotomorphogenesis is unaffected in fln seedlings. Seedlings genetically fln1-1 with dexamethasone-inducible FLN1-HA expression at germination are phenotypically indistinguishable from wild-type. Induction of FLN-HA after 24 hours of germination cannot rescue the mutant phenotype, indicating that the effects of loss of FLN are not always reversible. Examination of chloroplast gene expression in fln1-1 and fln2-1 by qRT-PCR reveals that transcripts of PEP-dependent genes were specifically reduced compared to NEP-dependent genes in both single mutants. CONCLUSIONS: Our results demonstrate that each FLN protein contributes to wild type growth, and acting additively are absolutely essential for plant growth and development.

Exploring the Functional Relationship between y-Type Thioredoxins and 2-Cys Peroxiredoxins in Arabidopsis Chloroplasts.

Thioredoxins (Trxs) are small, ubiquitous enzymes that catalyze disulphide-dithiol interchange in target enzymes. The large set of chloroplast Trxs, including f, m, x and y subtypes, use reducing equivalents fueled by photoreduced ferredoxin (Fdx) for fine-tuning photosynthetic performance and metabolism through the control of the activity of redox-sensitive proteins. Although biochemical analyses suggested functional diversity of chloroplast Trxs, genetic studies have established that deficiency in a particular Trx subtype has subtle phenotypic effects, leading to the proposal that the Trx isoforms are functionally redundant. In addition, chloroplasts contain an NADPH-dependent Trx reductase with a joint Trx domain, termed NTRC. Interestingly, Arabidopsis mutants combining the deficiencies of x- or f-type Trxs and NTRC display very severe growth inhibition phenotypes, which are partially rescued by decreased levels of 2-Cys peroxiredoxins (Prxs). These findings indicate that the reducing capacity of Trxs f and x is modulated by the redox balance of 2-Cys Prxs, which is controlled by NTRC. In this study, we explored whether NTRC acts as a master regulator of the pool of chloroplast Trxs by analyzing its functional relationship with Trxs y. While Trx y interacts with 2-Cys Prxs in vitro and in planta, the analysis of Arabidopsis mutants devoid of NTRC and Trxs y suggests that Trxs y have only a minor effect, if any, on the redox state of 2-Cys Prxs.

Chloroplast Redox Regulatory Mechanisms in Plant Adaptation to Light and Darkness.

Light is probably the most important environmental stimulus for plant development. As sessile organisms, plants have developed regulatory mechanisms that allow the rapid adaptation of their metabolism to changes in light availability. Redox regulation based on disulfide-dithiol exchange constitutes a rapid and reversible post-translational modification, which affects protein conformation and activity. This regulatory mechanism was initially discovered in chloroplasts when it was identified that enzymes of the Calvin-Benson cycle (CBC) are reduced and active during the day and become rapidly inactivated by oxidation in the dark. At present, the large number of redox-sensitive proteins identified in chloroplasts extend redox regulation far beyond the CBC. The classic pathway of redox regulation in chloroplasts establishes that ferredoxin (Fdx) reduced by the photosynthetic electron transport chain fuels reducing equivalents to the large set of thioredoxins (Trxs) of this organelle via the activity of a Fdx-dependent Trx reductase (FTR), hence linking redox regulation to light. In addition, chloroplasts harbor an NADPH-dependent Trx reductase with a joint Trx domain, termed NTRC. The presence in chloroplasts of this NADPH-dependent redox system raises the question of the functional relationship between NTRC and the Fdx-FTR-Trx pathways. Here, we update the current knowledge of these two redox systems focusing on recent evidence showing their functional interrelationship through the action of the thiol-dependent peroxidase, 2-Cys peroxiredoxin (2-Cys Prx). The relevant role of 2-Cys Prxs in chloroplast redox homeostasis suggests that hydrogen peroxide may exert a key function to control the redox state of stromal enzymes. Indeed, recent reports have shown the participation of 2-Cys Prxs in enzyme oxidation in the dark, thus providing an explanation for the long-lasting question of photosynthesis deactivation during the light-dark transition.

2-Cys Peroxiredoxins Participate in the Oxidation of Chloroplast Enzymes in the Dark.

Most redox-regulated chloroplast enzymes are reduced during the day and oxidized during the night. While the reduction mechanism of light-dependent enzymes is well known, the mechanism mediating their oxidation in the dark remains unknown. The thiol-dependent peroxidases, 2-Cys peroxiredoxins (Prxs), play a key role in light-dependent reduction of chloroplast enzymes. Prxs transfer reducing equivalents of thiols to hydrogen peroxide, suggesting the participation of these peroxidases in enzyme oxidation in the dark. Here, we have addressed this issue by analyzing the redox state of well-known redox-regulated chloroplast enzymes in response to darkness in Arabidopsis thaliana mutants deficient in chloroplast-localized Prxs (2-Cys Prxs A and B, Prx IIE, and Prx Q). Mutant plants lacking 2-Cys Prxs A and B, and plants overexpressing NADPH-dependent thioredoxin (Trx) reductase C showed delayed oxidation of chloroplast enzymes in the dark. In contrast, the deficiencies of Prx IIE or Prx Q exerted no effect. In vitro assays allowed the reconstitution of the pathway of reducing equivalents from reduced fructose 1,6-bisphosphatase to hydrogen peroxide mediated by Trxs and 2-Cys Prxs. Taken together, these results suggest that 2-Cys Prxs participate in the short-term oxidation of chloroplast enzymes in the dark.

An event of alternative splicing affects the expression of the NTRC gene, encoding NADPH-thioredoxin reductase C, in seed plants.

The NTRC gene encodes a NADPH-dependent thioredoxin reductase with a joint thioredoxin domain, exclusive of photosynthetic organisms. An updated search shows that although most species harbor a single copy of the NTRC gene, two copies were identified in different species of the genus Solanum, Glycine max and the moss Physcomitrella patens. The phylogenetic analysis of NTRCs from different sources produced a tree with the major groups of photosynthetic organisms: cyanobacteria, algae and land plants, indicating the evolutionary success of the NTRC gene among photosynthetic eukaryotes. An event of alternative splicing affecting the expression of the NTRC gene was identified, which is conserved in seed plants but not in algae, bryophytes and lycophytes. The alternative splicing event results in a transcript with premature stop codon, which would produce a truncated form of the enzyme. The standard splicing/alternative splicing (SS/AS) transcripts ratio was higher in photosynthetic tissues from Arabidopsis, Brachypodium and tomato, in line with the higher content of the NTRC polypeptide in these tissues. Moreover, environmental stresses such as cold or high salt affected the SS/AS ratio of the NTRC gene transcripts in Brachypodium seedlings. These results suggest that the alternative splicing of the NTRC gene might be an additional mechanism for modulating the content of NTRC in photosynthetic and non-photosynthetic tissues of seed plants.

[Breastfeeding and cognitive development; interference evaluation by "5 digits test"]. / Lactancia materna y desarrollo cognitivo; valoración de la respuesta a la interferencia mediante el "test de los 5 dígitos"

BACKGROUND AND OBJECTIVE: Different publications have analyzed the possible correlation between breastfeeding length and a better cognitive development during school age. The present study evaluates the possible long term beneficial effects of breastfeeding during the first months of life on the cognitive development, as well as its possible role on the attention deficit and hyperactivity prevention. Currently this evaluation is of special interest considering the increasing number in western societies of childhood attention deficits, with or without, hyper - activity disorders. MATERIAL AND METHODS: 103 children, during the first year of elementary education, 6 years of age (47 boys and 56 girls), were recruited in different schools of the Granada province, including urban, semi-urban and rural areas. The speed of cognitive processing and the ability to focus attention and reorient after an incident of distraction, using the five digits test (5-DGT), - variant of the Stroop interference test- were analyzed. RESULTS: Data show a linear correlation between longer breastfeeding and better scores in the test. Particularly significant (P ≤ 0.001) are the reading and alternation tests for children who were breastfed for 6 months when compared with those that only received this nutrition during their first month of life. CONCLUSION: This study validates the initial hypothesis, showing faster resolution speed and less interference in the group of children breastfed, at least during the first 6 months of life. Further well designed studies are necessary for reinforced breastfeeding recommendation, at least during the first 6 months of life, for children underclinical suspicion of possible ADHD development.

Introducción y objetivo: Numerosos estudios han intentado demostrar que la duración de la lactancia materna se relaciona con un mejor desarrollo cognitivo en la edad escolar. En el presente estudio se evalúa el potencial efecto beneficioso a largo plazo de la alimentación con leche materna durante los primeros meses de vida, no sólo sobre el desarrollo cognitivo, sino también como posible prevención del déficit de atención e hiperactividad. Esta valoración resulta de especial interés en la sociedad actual en la que se han incrementado de forma muy notable los déficits de atención en la infancia, unidos o no a trastornos de hiperactividad. Material y Métodos: Un total de 103 niños, escolarizados en primer curso de Educación Primaria, 6 años de edad, (47 niños y 56 niñas), fueron reclutados en diferentes colegios de la provincia de Granada de áreas urbanas, semiurbanas y rurales. Se analizó la velocidad de procesamiento cognitivo y la capacidad de enfocar la atención y reorientarla tras un suceso de distracción, mediante el test de los cinco dígitos (5-DGT), variante del test de interferencia de Stroop. Resultados: Los datos demuestran una correlación lineal entre una mayor duración de la lactancia materna y mejores resultados en todas las pruebas del test. Resultan altamente significativas (P ≤0,001) las comparaciones para las pruebas de lectura y alternancia, entre aquellos niños que fueron alimentados con leche materna durante 6 meses frente a los que sólo recibieron esta alimentación durante su primer mes de vida. Conclusión: El estudio valida la hipótesis inicial, demostrando una mayor velocidad de resolución y una menor interferencia en el grupo de niños alimentados con pecho al menos durante los 6 primeros meses. Los datos obtenidos deberían refrendarse en un posterior estudio, con una mayor muestra, ya que resulta de suma importancia reforzar el consejo de lactancia materna durante al menos los 6 primeros meses de vida ante neonatos con sospecha clínica de posible desarrollo de TDAH.

A proposed reaction mechanism for rice NADPH thioredoxin reductase C, an enzyme with protein disulfide reductase activity.

NADPH thioredoxin reductase C (NTRC) is an interesting NTR with a thioredoxin (Trx) domain at the C-terminus, able to conjugate both activities for 2-Cys peroxiredoxin (Prx) reduction. NTRC is dimeric in the presence of NADPH and interacted with dimeric 2-Cys Prx through the Trx module by a mixed disulfide between Cys377 of NTRC and Cys61 of the 2-Cys Prx. NTRC variants of both NTR and Trx active sites were inactive, but 1:1 mixtures of both variants allowed partial recovery of activity suggesting inter-subunit transfer of electrons during catalysis. Based on these results we propose a model for the reaction mechanism of NTRC.

The quaternary structure of NADPH thioredoxin reductase C is redox-sensitive.

NADPH thioredoxin reductase C (NTRC) is a chloroplast enzyme able to conjugate NADPH thioredoxin reductase (NTR) and thioredoxin (TRX) activities for the efficient reduction of 2-Cys peroxiredoxin (2-Cys PRX). Because NADPH can be produced in chloroplasts during darkness, NTRC plays a key role for plant peroxide detoxification during the night. Here, it is shown that the quaternary structure of NTRC is highly dependent on its redox status. In vitro, most of the enzyme adopted an oligomeric state that disaggregated in dimers upon addition of NADPH, NADH, or DTT. Gel filtration and Western blot analysis of protein extracts from Arabidopsis chloroplast stroma showed that native NTRC forms aggregates, which are sensitive to NADPH and DTT, suggesting that the aggregation state might be a significant aspect of NTRC activity in vivo. Moreover, the enzyme is localized in clusters in Arabidopsis chloroplasts. NTRC triple and double mutants, A164G-V182E-R183F and A164G-R183F, replacing key residues of NADPH binding site, showed reduced activity but were still able to dimerize though with an increase in intermediary forms. Based on these results, we propose that the catalytically active form of NTRC is the dimer, which formation is induced by NADPH.

Lactancia materna y desarrollo cognitivo; valoración de la respuesta a la interferencia mediante el "test de los 5 dígitos" / Breastfeeding and cognitive development; interference evaluation by "5 digits test"

Introducción y objetivo: Numerosos estudios han intentado demostrar que la duración de la lactancia materna se relaciona con un mejor desarrollo cognitivo en la edad escolar. En el presente estudio se evalúa el potencial efecto beneficioso a largo plazo de la alimentación con leche materna durante los primeros meses de vida, no sólo sobre el desarrollo cognitivo, sino también como posible prevención del déficit de atención e hiperactividad. Esta valoración resulta de especial interés en la sociedad actual en la que se han incrementado de forma muy notable los déficits de atención en la infancia, unidos o no a trastornos de hiperactividad. Material y Métodos: Un total de 103 niños, escolarizados en primer curso de Educación Primaria, 6 años de edad, (47 niños y 56 niñas), fueron reclutados en diferentes colegios de la provincia de Granada de áreas urbanas, semiurbanas y rurales. Se analizó la velocidad de procesamiento cognitivo y la capacidad de enfocar la atención y reorientarla tras un suceso de distracción, mediante el test de los cinco dígitos (5-DGT), variante del test de interferencia de Stroop. Resultados: Los datos demuestran una correlación lineal entre una mayor duración de la lactancia materna y mejores resultados en todas las pruebas del test. Resultan altamente significativas (P ≤ 0,001) las comparaciones para las pruebas de lectura y alternancia, entre aquellos niños que fueron alimentados con leche materna durante 6 meses frente a los que sólo recibieron esta alimentación durante su primer mes de vida. Conclusión: El estudio valida la hipótesis inicial, demostrando una mayor velocidad de resolución y una menor interferencia en el grupo de niños alimentados con pecho al menos durante los 6 primeros meses. Los datos obtenidos deberían refrendarse en un posterior estudio, con una mayor muestra, ya que resulta de suma importancia reforzar el consejo de lactancia materna durante al menos los 6 primeros meses de vida ante neonatos con sospecha clínica de posible desarrollo de TDAH (AU)

Background and objective: Different publications have analyzed the possible correlation between breastfeeding length and a better cognitive development during school age. The present study evaluates the possible long term beneficial effects of breastfeeding during the first months of life on the cognitive development, as well as its possible role on the attention deficit and hyperactivity prevention. Currently this evaluation is of special interest considering the increasing number in western societies of childhood attention deficits, with or without, hyper - activity disorders. Material and methods: 103 children, during the first year of elementary education, 6 years of age (47 boys and 56 girls), were recruited in different schools of the Granada province, including urban, semi-urban and rural areas. The speed of cognitive processing and the ability to focus attention and reorient after an incident of distraction, using the five digits test (5-DGT), - variant of the Stroop interference test- were analyzed. Results: Data show a linear correlation between longer breastfeeding and better scores in the test. Particularly significant (P ≤ 0.001) are the reading and alternation tests for children who were breastfed for 6 months when compared with those that only received this nutrition during their first month of life. Conclusion: This study validates the initial hypothesis, showing faster resolution speed and less interference in the group of children breastfed, at least during the first 6 months of life. Further well designed studies are necessary for reinforced breastfeeding recommendation, at least during the first 6 months of life, for children underclinical suspicion of possible ADHD development (AU)

Lactancia materna y su influencia en los procesos cognitivos de escolares españoles (6 años de edad), valorada con la escala de inteligencia Wechsler / Breastfeeding and its influence into the cognitive process of Spanish school-children (6 years old), measured by the Wechsler intelligence scale

Numerosos estudios han intentado demostrar que la duración de la lactancia materna se relaciona con un mejor desarrollo cognitivo en la edad escolar. En el presente estudio se evalúa el potencial efecto beneficioso a largo plazo de la alimentación con leche materna durante los primeros meses de vida, mejorando el desarrollo cognitivo y aquellas funciones relacionadas con la Comprensión Verbal, Razonamiento Perceptivo, Memoria de Trabajo y Velocidad de Procesamiento. Un total de 103 niños, escolarizados en primer curso de Educación Primaria, 6 años de edad, (47 niños y 56 niñas), en diferentes colegios de la provincia de Granada, en áreas urbanas, semiurbanas y rurales. Se evaluó la capacidad cognitiva global y algunos dominios específicos de inteligencia a través de la escala de inteligencia Wechsler para niños -IV. Los resultados demuestran una asociación estadísticamente significativa, entre las mejores puntuaciones en el cociente intelectual (CI) y en los cuatro índices medidos mediante el test WISC-IV, con una mayor duración de la lactancia. Resulta altamente significativas (p=0,000) la asociación entre las mejores puntuaciones en el test y 6 meses de lactancia, lo que valida nuestra hipótesis. Como medio de prevención de diferentes déficit intelectuales se debe reforzar el consejo de lactancia materna exclusiva durante al menos los 6 primeros meses de vida.

Some scientific evidence support that a better cognitive development during the school age is related with breastfeeding. In this study, the potential benefit of breastfeeding duration is evaluated, related to Verbal Comprehension, Perceptual Reasoning, Working Memory and Processing Speed. A total of 103 children, first year of Primary School, six-year-old, (47 boys and 56 girls), were included from different schools in the province of Granada (Spain) at urban, semi-urban and rural areas. The global cognitive capability, as well as some specific intelligence domains which permit a more precise and deeper analysis of the cognitive processes, was evaluated through the Wechsler Intelligence Scale for Children - version IV. The results prove an association, statistically signnificative, between the best values of IQ and the other four WISC-IV indexes and a longer breastfeeding. There is a highly significant (p=0,000) association between the best scores and those children who were breastfed during 6 months, which validates our hypothesis. The advice of breastfeeding during at least the first six months of life should be reinforced to reduce learning difficulties.

Rice NTRC is a high-efficiency redox system for chloroplast protection against oxidative damage.

One of the mechanisms plants have developed for chloroplast protection against oxidative damage involves a 2-Cys peroxiredoxin, which has been proposed to be reduced by ferredoxin and plastid thioredoxins, Trx x and CDSP32, the FTR/Trx pathway. We show that rice (Oryza sativa) chloroplast NADPH THIOREDOXIN REDUCTASE (NTRC), with a thioredoxin domain, uses NADPH to reduce the chloroplast 2-Cys peroxiredoxin BAS1, which then reduces hydrogen peroxide. The presence of both NTR and Trx-like domains in a single polypeptide is absolutely required for the high catalytic efficiency of NTRC. An Arabidopsis thaliana knockout mutant for NTRC shows irregular mesophyll cell shape, abnormal chloroplast structure, and unbalanced BAS1 redox state, resulting in impaired photosynthesis rate under low light. Constitutive expression of wild-type NTRC in mutant transgenic lines rescued this phenotype. Moreover, prolonged darkness followed by light/dark incubation produced an increase in hydrogen peroxide and lipid peroxidation in leaves and accelerated senescence of NTRC-deficient plants. We propose that NTRC constitutes an alternative system for chloroplast protection against oxidative damage, using NADPH as the source of reducing power. Since no light-driven reduced ferredoxin is produced at night, the NTRC-BAS1 pathway may be a key detoxification system during darkness, with NADPH produced by the oxidative pentose phosphate pathway as the source of reducing power.