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Publisher Summary This chapter focuses on the importance of uptake and release of elements by leaves and other aerial parts and discusses the practical aspects of foliar fertilization and the ecological consequences of nutrient uptake and release. The surface of plants is covered by the cuticle, which is a bio-polymer synthesized by epidermal cells and primarily consists of cutin, a polyester matrix of polymerized long-chain fatty acids in which waxes are embedded. It protects the leaf from excessive water loss by transpiration and against excessive leaching of inorganic and organic solutes by rain. The hydrophobic nature of the cuticle makes it an effective barrier against the penetration by hydrophilic polar solutes, whereas lipophilic molecules may penetrate cuticles at much higher rates. Stomata act as adjustable apertures in the leaf surface, which optimize the trade-off between CO − uptake and water loss of plants. Various environmental conditions during plant growth, such as shading, high temperatures, and humidity, as well as nutrient deficiency, affect the structure and anatomy of the leaf surface, thereby affecting the uptake and release of elements by the leaves. Foliar sprays are widely used in agricultural production as an alternative or complementary strategy to soil fertilization, and the plant responses to elements supplied via foliar sprays are normally more rapid than to soil treatment. Thus it can help in preserving crop yields and quality, with low environmental impact.

Objective To estimate the SARS-CoV-2 antibody seroprevalence in healthcare workers (HCWs) at a university hospital in Mallorca, Spain. Methods All HCWs received an e-mail inviting them to take part in the study. Participants had a nasopharyngeal swab test performed for reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and serological tests to detect SARS-CoV-2 antibodies (primary study). Additionally, they were invited to complete a questionnaire on their exposure to COVID-19 individuals and their COVID-19-related symptoms (secondary study). Prevalence of antibodies (IgG, IgM, or both) and 95% confidence intervals (CIs) were calculated. Results Seventy-nine percent of the hospitalʼs HCWs (N = 2210) took part in the primary study. Antibodies were detected in 61 participants, a prevalence of 2.8% (95% CI: 2.5–3.1). The prevalence was slightly higher in nurses (3.4%), registrars (3.9%), and wardens (3.4%). Thirty-nine percent of the primary study participants completed the secondary study questionnaire. Those with positive antibody test results had closer contact with COVID-19 individuals (60% vs. 92%; p Conclusion After the first wave of the COVID-19 pandemic in Spain, the seroprevalence of SARS-CoV-2 antibodies in our university hospital HCWs was around 2.8%, which is slightly higher than the seroprevalence in the general population in our region. We believe it would be advisable to perform additional seroprevalence studies during the second wave of the epidemic.

The absorption of water and solutes by plant leaves has been recognised since more than two centuries. Given the polar nature of water and solutes, the mechanisms of foliar uptake have been proposed to be similar for water and electrolytes, including nutrient solutions. Research efforts since the XIX Century focussed on characterising the properties of cuticles and applying foliar sprays to crop plants as tool for improving crop nutrition. This was accompanied by the development of hundreds of studies aimed at characterising the chemical and structural nature of plant cuticles from different species and the mechanisms of cuticular and, to a lower extent, stomatal penetration of water and solutes. The processes involved are complex and will be affected by multiple environmental, physico-chemical and physiological factors which are only partially clear to date. During the last decades, there is growing evidence that water transport across leaf surfaces of native species may contribute to water balances (absorption and loss) at an ecosystem level. Given the potential importance of foliar water absorption for many plant species and ecosystems as shown in recent studies, the aim of this review is to first integrate current knowledge on plant surface composition, structure, wettability and physico-chemical interactions with surface-deposited matter. The different mechanisms of foliar absorption of water and electrolytes and experimental procedures for tracing the uptake process are discussed before posing several outstanding questions which should be tackled in future studies. Published

The cuticle is a lipid-rich layer that protects aerial plant organs against multiple stress factors such as dehydration. In this study, cuticle composition and structure in relation to water loss are examined in a broad ecophysiological context, taking into consideration leaf age and side from Ilex aquifolium (holly) in comparison with Eucalyptus globulus (eucalypt) and Prunus laurocerasus (cherry laurel). Enzymatically isolated cuticular membranes from holly leaves were studied under three treatment conditions: natural (no chemical treatment), after dewaxing, and after methanolysis, and the rate of water loss was assessed. Structural and chemical changes were evaluated using different microscopy techniques and by Fourier transform infrared (FTIR) spectroscopy. The potential mechanisms of solute absorption by holly leaves were additionally evaluated, also testing if its prickly leaf margin may facilitate uptake. The results indicate that the treatment conditions led to structural changes, and that chemical composition was hardly affected because of the occurrence of cutan. Structural changes led to more hydrophilic adaxial surfaces, which retained more water and were more efficient than natural cuticles, while changes were not significant for abaxial surfaces. Across natural cuticles, age was a significant factor for eucalypt but not for holly. Young eucalypt cuticles were the group that absorbed more water and had the lowest water loss rate. When comparing older leaf cuticles of the three species, cherry laurel was found to absorb more water, which was, however, lost more slowly, compared with the other species. Evidence was gained that holly leaves can absorb foliar-applied solutes (traced after calcium chloride application) through the adaxial and abaxial surfaces, the adaxial mid veins, and to a lower extent, the spines. In conclusion, for the species examined, the results show variations in leaf cuticle composition and structure in relation to leaf ontogeny, and water sorption and desorption capacity.

Developing neurons form synapses at a high rate. Synaptic transmission is very energy-demanding and likely requires ATP production by mitochondria nearby. Mitochondria might be targeted to active synapses in young dendrites, but whether such motility regulation mechanisms exist is unclear. We investigated the relationship between mitochondrial motility and neuronal activity in the primary visual cortex of young mice in vivo and in slice cultures. During the first 2 postnatal weeks, mitochondrial motility decreases while the frequency of neuronal activity increases. Global calcium transients do not affect mitochondrial motility. However, individual synaptic transmission events precede local mitochondrial arrest. Pharmacological stimulation of synaptic vesicle release, but not focal glutamate application alone, stops mitochondria, suggesting that an unidentified factor co-released with glutamate is required for mitochondrial arrest. A computational model of synaptic transmission-mediated mitochondrial arrest shows that the developmental increase in synapse number and transmission frequency can contribute substantially to the age-dependent decrease of mitochondrial motility.

There is still around 50% of the familial breast cancer (BC) cases with an undefined genetic cause, here we have used next-generation sequencing (NGS) technology to identify new BC susceptibility genes. This approach has led to the identification of RECQL5, a member of RECQL-helicases family, as a new BC susceptibility candidate, which deserves further study. We have used a combination of whole exome sequencing in a family negative for mutations in BRCA1/2 throughout (BRCAX), in which we found a probably deleterious variant in RECQL5, and targeted NGS of the complete coding regions and exon-intron boundaries of the candidate gene in 699 BC Spanish BRCAX families and 665 controls. Functional characterization and in silico inference of pathogenicity were performed to evaluate the deleterious effect of detected variants. We found at least seven deleterious or likely deleterious variants among the cases and only one in controls. These results prompt us to propose RECQL5 as a gene that would be worth to analyze in larger studies to explore its possible implication in BC susceptibility.

© 2020 by the authors., Background: Tumor-associated macrophages (TAMs) play a crucial role in suppressing the immunosurveillance function of the immune system that prevents tumor growth. Indeed, macrophages can also be targeted by different chemotherapeutic agents improving the action over immune checkpoints to fight cancer. Here we describe the effect of trabectedin and lurbinectedin on human macrophage cell viability and function. Methods: Blood monocytes from healthy donors were differentiated into macrophages and exposed to different stimuli promoting functional polarization and differentiation into tumor-associated macrophages. Cells were challenged with the chemotherapeutic drugs and the effects on cell viability and function were analyzed. Results: Human macrophages exhibit at least two different profiles in response to these drugs. One-fourth of the blood donors assayed (164 individuals) were extremely sensitive to trabectedin and lurbinectedin, which promoted apoptotic cell death. Macrophages from other individuals retained viability but responded to the drugs increasing reactive oxygen production and showing a rapid intracellular calcium rise and a loss of mitochondrial oxygen consumption. Cell-membrane exposure of programmed-death ligand 1 (PD-L1) significantly decreased after treatment with therapeutic doses of these drugs, including changes in the gene expression profile of hypoxia-inducible factor 1 alpha (HIF-1α)-dependent genes, among other. Conclusions: The results provide evidence of additional onco-therapeutic actions for these drugs., This research was supported by the Ministerio Economía, Industria y Competitividad (SAF2017-82436-R/RTC2017-6283-1 and PID2019-108674RB-100), Centro de Investigación Biomédica en Red en Enfermedades Cardiovasculares (CB16/11/00222), PharmaMar (contract 164249), Fundación Ramón Areces (CIUP18A3864), Consorcio de Investigación en Red de la Comunidad de Madrid, S2017/BMD-3686 and Fondo Europeo de Desarrollo Regional.

Growing conditions at different tree canopy positions may significantly vary and lead to foliar changes even within the same tree. An assessment of foliar anatomy, including also epidermal features, can help us understand how plants respond to environmental factors. Working with two model tree species (i.e., Quercus petraea and Fagus sylvatica) grown at their southernmost European distribution area in Central Spain, the influence of irradiation and canopy height was examined by sampling lower canopy leaves and comparing them with fully irradiated, top canopy leaves and shaded top canopy leaves grown for months within a bag made of shade netting fabric before they sprouted. At the end of the summer, samples were collected, and several parameters were analysed. The results indicate that SLA (specific leaf area) differences are significant both between species and groups. Leaf and cuticle thickness differed significantly between groups while stomatal densities only between species. Regarding mineral concentrations, differences between species were significant for K, Mn, N and N: P ratios. It is concluded that leaf responses to environmental conditions may be variable both within the same tree and between species.

A 90-d feeding trial was conducted in which five groups of gilthead seabream (11.96 g initial body weight) were fed with a microalgae-free diet (control group, C) or four diets containing the microalgae Nannochloropsis gaditana at two inclusion levels (2.5% or 5%), either raw (R2.5 and R5 batches) or cellulose-hydrolyzed (H2.5 and H5 batches), to study their effect on the body and muscle growth. At 40 days, the highest values of body length and weight were reached in R5 group, but at 64 and 90 days, these were reached in R2.5. However, feed conversion rate, specific growth, daily intake, and survival (100%) were similar in all the groups. The acquisition of a discoid body shape was accelerated depending on the inclusion level of N. gaditana in the diets. Moreover, H5 diet affected the fish geometric morphology compared to R5 diet. The white muscle transverse area was similar in all groups at 40 days, with the exception of H2.5 group, which showed the lowest area. At day 90, C and R2.5 displayed the highest muscle growth, attributable to increased hyperplasia in C, and higher hypertrophy in R2.5. However, the highest proportion of small and medium fibers was observed in R5 and H5, ALGAFISH, Evaluación de los efectos de las microalgas, incluidas en dietas de dorada, sobre el crecimiento, supervivencia y resistencia a infecciones., SI

The contribution of the nucleotide-binding oligomerization domain protein NOD1 to obesity has been investigated in mice fed a high fat diet (HFD). Absence of NOD1 accelerates obesity as early as 2 weeks after feeding a HFD. The obesity was due to increases in abdominal and inguinal adipose tissues. Analysis of the resting energy expenditure showed an impaired function in NOD1-deficient animals, compatible with an alteration in thyroid hormone homeostasis. Interestingly, free thyroidal T4 increased in NOD1-deficient mice fed a HFD and the expression levels of UCP1 in brown adipose tissue were significantly lower in NOD1-deficient mice than in the wild type animals eating a HFD, thus contributing to the observed adiposity in NOD1-deficient mice. Feeding a HFD resulted in an alteration of the proinflammatory profile of these animals, with an increase in the infiltration of inflammatory cells in the liver and in the white adipose tissue, and an elevation of the circulating levels of TNF-α. In addition, alterations in the gut microbiota in NOD1-deficient mice correlate with increased vulnerability of their ecosystem to the HFD challenge and affect the immune-metabolic phenotype of obese mice. Together, the data are compatible with a protective function of NOD1 against low-grade inflammation and obesity under nutritional conditions enriched in saturated lipids. Moreover, one of the key players of this early obesity onset is a dysregulation in the metabolism and release of thyroid hormones leading to reduced energy expenditure, which represents a new role for these hormones in the metabolic actions controlled by NOD1., This work was supported by Grants SAF2017-82436R, AGL2017-88801-P and SAF2016-75004R from MINECO/AEI/FEDER/EU, S2017/BMD-3686 from Comunidad de Madrid, CIVP18A3864 from Fundación Ramón Areces and CIBERCV and CIBERHED (funded by the Instituto de Salud Carlos III) and Fondos FEDER.

Nutrient resorption is crucial for mineral element conservation and efficiency of forest species, but knowledge on its significance and the mechanisms involved is still limited for most species and habitats. Focusing on the harsh conditions for plant growth and survival of southern Patagonia, a field study for comparing the rate of foliar resorption of macro-, micro-nutrients, and trace elements in coexisting Nothofagus pumilio and Nothofagus antarctica forests was performed. Forests located in three contrasting productivity sites (with different soil and climatic conditions) were selected, and mature, functional versus senescent leaves of both species were collected at two different dates of the growing season. Macro- (N, P, Ca, K, S, and Mg), micronutrients (B, Cu, Fe, Mn, Zn, and Ni), and trace elements (Al, Li, Pb, Rb, Sr, Ti, and Tl) were determined in foliar tissues. The mineral element concentrations of mature and senescent leaves were used for calculating the nutrient resorption efficiency (NuR). In general, and making an average of all sites and species, macro-nutrient resorption showed a decreasing trend for N > S = K > P > Mg, being Ca the only macro-nutrient with negative values (i.e., no resorption). Resorption of the majority of the elements did not vary between species in any of the evaluated sites. Variation across sites in nutrient resorption efficiency for most macronutrients, some micronutrients, and trace elements was observed for N. antarctica, whereas N. pumilio had a similar NuR for all experimental sites. On the other hand, regardless of the site or the species, some elements were not resorbed (e.g., B, Cu, Fe, Mn, Al, and Ti). It is concluded that both Nothofagus species performed similarly concerning their nutrient conservation strategy, when coexisting in the same mixed forest. However, no evidence was gained for an increased rate of foliar NuR in association with the sites subjected to more limiting soil and climatic conditions for plant growth. EEA Santa Cruz Fil: Bahamonde, Héctor Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Universidad Nacional de la Patagonia Austral; Argentina Fil: Fernández, Victoria. Universidad Tecnológica de Madrid. School of Forest Engineering. Forest Genetics and Ecophysiology Research Group; España Fil: Gyenge, Javier. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Agencia de Extensión Rural Tandil; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mattenet, Francisco Javier. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina Fil: Peri, Pablo Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Universidad Nacional de la Patagonia Austral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

© 2021 The Author(s)., Nucleotide-binding oligomerization domain 1 (NOD1), a pattern recognition receptor (PRR) that detects bacterial peptidoglycan fragments and other danger signals, has been linked to inflammatory pathologies. NOD1, which is expressed by immune and non-immune cells, is activated after recognizing microbe-associated molecular patterns (MAMPs). This recognition triggers host defense responses and both immune memory and tolerance can also be achieved during these processes. Since the gut microbiota is currently considered a master regulator of human physiology central in health and disease and the intestine metabolizes a wide range of nutrients, drugs and hormones, it is a fact that dysbiosis can alter tissues and organs homeostasis. These systemic alterations occur in response to gastrointestinal immune adaptations that are not yet fully understood. Even if previous evidence confirms the connection between the microbiota, the immune system and metabolic disorders, much remains to be discovered about the contribution of NOD1 to low-grade inflammatory pathologies such as obesity, diabetes and cardiovascular diseases. This review compiles the most recent findings in this area, while providing a dynamic and practical framework with future approaches for research and clinical applications on targeting NOD1. This knowledge can help to rate the consequences of the disease and to stratify the patients for therapeutic interventions., The review was written by authors that work in a laboratory where projects are supported by: Ministerio de Economía, Industria y Competitividad/Agencia Estatal de Investigación 10.13039/501100011033 (RTI2018-094727-B100, SAF2017-82436-R, BIO2016-77639-P, PID2019-108977-RBI00 and PID2020-113238-RBI00), Centro de Investigación Biomédica en Red en Enfermedades Cardiovasculares (CB16/11/00222), Consorcio de Investigación en Red de la Comunidad de Madrid, S2017/BMD-3686 and Fondo Europeo de Desarrollo Regional.

Plant cuticles have been traditionally classified on the basis of their ultrastructure, with certain chemical composition assumptions. However, the nature of the plant cuticle may be misinterpreted in the prevailing model, which was established more than 150 years ago. Using the adaxial leaf cuticle of Ficus elastica, a study was conducted with the aim of analyzing cuticular ultrastructure, chemical composition and the potential relationship between structure and chemistry. Gradual chemical extractions and diverse analytical and microscopic techniques were performed on isolated leaf cuticles of two different stages of development (i.e. young and mature leaves). Evidence for the presence of cutan in F. elastica leaf cuticles has been gained after chemical treatments and tissue analysis by infrared spectroscopy and electron microscopy. Significant calcium, boron and silicon concentrations were also measured in the cuticle of this species. Such mineral elements which are often found in plant cell walls may play a structural role and their presence in isolated cuticles further supports the interpretation of the cuticle as the most external region of the epidermal cell wall. The complex and heterogeneous nature of the cuticle, and constraints associated with current analytical procedures may limit the chance for establishing a relationship between cuticle chemical composition and structure also in relation to organ ontogeny.

Plant surfaces have a considerable degree of chemical and physical variability also in relation to different environmental conditions, organs and state of development. The potential changes on plant surface properties in association with environmental variations have been little explored so far. Using two model tree species (i.e., Quercus petraea, sessile oak and Fagus sylvatica, beech) growing in ‘Montejo de la Sierra Forest,’ we examined various traits of the abaxial and adaxial surface of leaves of both species collected at a height of approximately 15 m (top canopy), versus 3.5–5.5 m for beech and sessile oak, lower canopy leaves. Leaf surface ultra-structure was analyzed by scanning and transmission electron microscopy, and the surface free energy and related parameter were estimated after measuring drops of 3 liquids with different degrees of polarity and apolarity. The permeability of the adaxial and abaxial surface of top and bottom canopy leaves to CaCl2 was estimated by depositing 2 drops of 3–4 ml per cm2 and comparing the concentration of Ca in leaf tissues 24 h after treatment, and also Ca and Cl concentrations in the washing liquid. Higher Ca concentrations were recorded after the application of CaCl2 drops onto the veins and adaxial blade of top canopy beech leaves, while no significant evidence for foliar Ca absorption was gained with sessile oak leaves. Surprisingly, high amounts of Cl were recovered after washing untreated, top canopy beach and sessile oak leaves with deionised water, a phenomenon which was not traced to occur on lower canopy leaves of both species. It is concluded that the surface of the two species analyzed is heterogeneous in nature and may have areas favoring the absorption of water and solutes as observed for the veins of beech leaves. Fil: Bahamonde, Héctor Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Universidad Nacional de la Patagonia Austral; Argentina Fil: Fernández, Victoria. Universidad Politécnica de Madrid. Escuela Técnica Superior de Ingeniería de Montes, Forestal y del Medio Natural. Departamento de Sistemas y Recursos Naturales; España Fil: Gil, Luis. Universidad Politécnica de Madrid. Escuela Técnica Superior de Ingeniería de Montes, Forestal y del Medio Natural. Departamento de Sistemas y Recursos Naturales; España

Fruits may be exposed to several unfavorable mechanical, climatic, and biological factors during ripening. Harvest and storage also challenge fruit integrity before fruits reach the consumer. In order to preserve fruit properties/characteristics it is essential that the structural and chemical integrity of the cuticle is maintained throughout fruit development and expansion. In addition, cuticles serve as protection against multiple biotic and abiotic stress factors and primarily act as a barrier to prevent water loss. Despite the important functions attributed to the cuticle, little is known about fruit cuticle biosynthesis and assembly, which is highly relevant when coping with adverse conditions. Presently, drought and heat pose severe constraints to the fruit industry via penalties in yield and fruit quality. Available climate change models suggest a scenario in which the impact of these environmental factors will negatively affect the fruit industry. A comprehensive understanding of the physiological and biochemical effects of limited water availability on fruit traits is a prerequisite for implementing breeding and knowledge-based strategies that enhance fruit crop tolerance to limited water availability. To address some of these questions, this chapter aimed to revise the existing information on cuticle physiology, composition, structure, and properties, also considering its impact on fruit under abiotic stresses, with an emphasis on water deficit. We also address the recent molecular progress in cuticle biosynthesis pathways and highlight some of the major research questions that will have to be dealt with in the future.

ResumenEn la actualidad se conocen 8.000 enfermedades genéticas monogénicas. La mayoría de ellas son heterogéneas, por lo que el diagnóstico molecular por técnicas convencionales de secuenciación suele ser largo y costoso debido al gran número de genes implicados. El tiempo estimado para el diagnóstico molecular se encuentra entre 1 y 10 años, y este retraso impide que los pacientes reciban medidas terapéuticas y de rehabilitación específicas, que sus familiares entren en programas preventivos y que reciban asesoramiento genético.La secuenciación masiva está cambiando el modelo de diagnóstico molecular de los afectos, sin embargo, los médicos y profesionales de la salud se enfrentan al dilema de la selección del método más eficiente, con el menor coste sanitario y con la mayor precisión de sus resultados. El objetivo de este trabajo es revisar la tecnología de secuenciación masiva y definir las ventajas y los problemas en su utilización.SummaryCurrently 8000 monogenic genetic diseases are known. Most of them are heterogeneous, so their molecular diagnosis by conventional sequencing techniques is labour intensive and time consuming due to the large number of genes involved. The estimated time is between 1 and 10 years for molecular diagnosis and this delay prevents patients from receiving therapy and rehabilitation measures, and their families from entering prevention programs and being given genetic counselling.Next generation sequencing (NGS) is changing the model of molecular diagnosis of patients; however, doctors and health professionals are faced with the dilemma of choosing the most efficient method, with lower health care costs and the most accurate results. The aim of this paper is to review the NGS technology and define the advantages and problems in the use of this technology.

European white elm (Ulmus laevis Pallas) populations are scarce, small and fragmented in the Iberian Peninsula. Due to these characteristics the indigenous status of the species in the region has been questioned, whilst the species’ role in Iberian riparian forest ecology has been neglected. Herein we review past studies regarding this species’ distribution and ecology in the Iberian Peninsula, with special emphasis on the establishment of conservation priorities. We first present a collection of palaeogeographic, historic and genetic data suggesting that the Iberian Peninsula was a glacial refuge for U. laevis. Secondly, we analyse U. laevis distribution in relation to soil physico- chemical properties and water availability in Spain. Following this, we focus on the reproductive biology of the species, and investigate the effect of masting and empty seed production on predation and regeneration establishment. Finally, based on this knowledge, we propose conservation policies for U. laevis in the Iberian Peninsula.

Novel next-generation sequencing procedures have rapidly emerged into the preimplantation genetic screening framework. This work presents the design and validation of a new low-coverage whole-genome sequencing assay for aneuploidy detection in single blastomeres and trophectodermal samples from preimplantation embryos. The validation ensures analytical sensitivity, specificity, robustness, precision, limit of detection, resolution, and reproducibility. Specific parameters to measure the performance are defined, and the results are compared with a standardized array-based method to stablish the concordance. From the single cell genomics point of view, the main novelties are the length of reads of the libraries (150 nucleotides) together with a paired-end strategy and the design of an original algorithm and copy number viewer. A total of 129 samples were included in six experimental runs using a MiSeq Illumina platform. Samples included: single amniocytes, single blastomeres (cleavage-stage embryos), trophectoderm samples (blastocyst), and diluted DNA. Sensitivity and specificity were calculated per chromosome yielding 96% and 99%, respectively. The percentage of concordant samples was 98.2% and all of the aneuploid samples were confirmed. In conclusion, the validation yields highly reliable and reproducible results, representing an accurate and cost-effective strategy for the routine detection of aneuploidy in human embryos.

The POT1 protein binds and protects telomeres. Germline variants in the POT1 gene have recently been shown to be associated with risk of developing tumors in different tissues such as familial chronic lymphocytic leukemia, colorectal, glioma and melanoma tumors. Recently, we uncovered a variant in the POT1 gene (p.R117C) as causative of familial cardiac angiosarcomas (CAS) in Li-Fraumeni-like (LFL) syndrome families. Our in silico studies predicted that this protein had lost the ability to interact with TPP1 and single-stranded DNA. In vitro studies corroborated this prediction and showed that this lack of function leads to abnormally long telomeres. To better understand the POT1 gene and its role with tumorigenesis, we extended the study to LFL (with and without members affected with angiosarcomas (AS)) and sporadic AS and cardiac sarcomas. We found POT1 variants in the 20% of the families with members affected with AS and 10% of sporadic AS and sarcomas. In silico studies predicted that these new variants were damaging in the same manner as previously described for the POT1 p.R117C variants. The wide spectrum of variants in the POT1 gene leading to tumorigenesis in different tissues demonstrates its general importance. Study of the POT1 gene should be considered as routine diagnostic in these cancers.

Few data are available concerning the role of risk markers for Alzheimer's disease (AD) in progression to AD dementia among subjects with mild cognitive impairment (MCI). We therefore investigated the role of well-known AD-associated single-nucleotide polymorphism (SNP) in the progression from MCI to AD dementia. Four independent MCI data sets were included in the analysis: (a) the German study on Aging, Cognition and Dementia in primary care patients (n=853); (b) the German Dementia Competence Network (n=812); (c) the Fundació ACE from Barcelona, Spain (n=1245); and (d) the MCI data set of the Amsterdam Dementia Cohort (n=306). The effects of single markers and combined polygenic scores were measured using Cox proportional hazards models and meta-analyses. The clusterin (CLU) locus was an independent genetic risk factor for MCI to AD progression (CLU rs9331888: hazard ratio (HR)=1.187 (1.054-1.32); P=0.0035). A polygenic score (PGS1) comprising nine established genome-wide AD risk loci predicted a small effect on the risk of MCI to AD progression in APOE-ϵ4 (apolipoprotein E-ϵ4) carriers (HR=1.746 (1.029-2.965); P=0.038). The novel AD loci reported by the International Genomics of Alzheimer's Project were not implicated in MCI to AD dementia progression. SNP-based polygenic risk scores comprising currently available AD genetic markers did not predict MCI to AD progression. We conclude that SNPs in CLU are potential markers for MCI to AD progression.

Sand dune quarries are a location of common aggregate mining activity developed in coastal areas, especially in the southeast Buenos Aires province, Argentina. In this article, spontaneous plant development after extraction activity ceased was evaluated. Five areas (three quarried and two natural/conservation areas) were sampled for plant cover and composition as well as sediment characterization. Different indexes, principal component analysis, and cluster analyses were applied to compare the areas. The dominant families observed in four of the five areas were Asteraceae, Poaceae, and Cyperaceae, and most of the species are commonly found in sandy and humid soils and/or modified/anthropized ones. Percentages of plant cover increased with time because of the cessation of active aggregate extraction. Indexes and multivariate analyses showed that it was possible to distinguish quarried and natural areas based on composition and vegetation cover. The distribution of plant species among the four areas responded to the presence of mining activity, but it also responded to the topographical position and consequently the depth of the groundwater level. Besides these differences, the four areas shared many native species. The results might indicate that once the activity has ceased, quarried areas may spontaneously and quickly develop a plant community with some similarities to those present in the nonquarried areas. However, given that the extracting activity involves the removal of the soil, revegetation of this type of environment depends on the presence of natural areas in the surroundings, which can serve as a source of seeds and propagules for plant regeneration. Fil: Fernandez Montoni, Maria Victoria. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina Fil: Fernández Honaine, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina Fil: del Rio, Julio Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina

Trabectedin is more active in nucleotide excision repair (NER)-efficient and homologous recombination repair (HRR)-deficient cells. As up to 25% of sporadic breast tumors present somatic inactivation of the HRR pathway (BRCAness phenotype), we sought to characterize trabectedin effect in BRCA1-proficient and BRCA1-null breast cancer cell lines. We evaluated whether HRR and NER gene expression correlates with trabectedin sensitivity and explored the response predictive value of the CUL4A ubiquitin ligase, which ubiquitinates NER pathway members. We characterized trabectedin cytotoxicity, cell-cycle effects, and BRCA1, BRCA2, XRCC3, XPG, ERCC1, and CUL4A expression in 10 breast cancer cell lines. Gene expression and trabectedin sensitivity association were determined in cell lines. Survival assays after trabectedin treatment were conducted in CUL4A-silenced BRCA1-proficient and -deficient cells. Because of limited phase II clinical trials evaluating trabectedin efficacy in patients with breast cancer, we assessed CUL4A immunohistochemical staining in a retrospective series of 118 sarcomas from trabectedin-treated patients to validate in vivo our in vitro observations. In cell lines, greater trabectedin sensitivity was associated with higher CUL4A expression and lower BRCA1/ERCC5, BRCA1/CUL4A, and XRCC3/CUL4A expression ratios. In agreement, BRCA1-deficient CUL4A-knockdown cells presented higher cell survival after trabectedin exposure than did scramble control cells. Lack of effect in BRCA1-proficient cells suggests that HRR impairment is key in CUL4A-mediated trabectedin sensitivity. High CUL4A expression in nontranslocation-related patients with sarcoma predicted improved progression-free survival [PFS; HR, 0.37; 95% confidence interval (CI), 0.20–0.68, P = 0.001] and overall survival (OS; HR, 0.44; 95% CI, 0.21–0.93, P = 0.026). Our observations support the notion of greater trabectedin activity in tumors exhibiting BRCAness and reveal CUL4A as a potential biomarker for definition of trabectedin target patients. Mol Cancer Ther; 12(4); 530–41. ©2013 AACR.

Plant surface properties influence solid-liquid interactions and matter exchange between the organs and their surrounding environment. In the case of fruits, surface processes may be of relevance for seed production and dispersal. To gain insight into the relationship between surface structure, chemical composition and function of aerial reproductive organs, we performed diverse experiments with the dry, winged fruits, or samaras, of Ulmus laevis Pall. and Ulmus minor Mill. both at the time of full maturity (green samaras) and of samara dispersal (dry samaras). Samaras of both elm species showed positive photosynthetic rates and absorbed water through their epidermal surfaces. The surface wettability, free energy, polarity and solubility parameter were lower in U. laevis than in U. minor and decreased for dry samaras in both species. Ulmus laevis samaras had a high degree of surface nano-roughness mainly conferred by cell wall folds containing pectins that substantially increased after hydration. The samaras in this species also had a thicker cuticle that could be isolated by enzymatic digestion, whereas that of U. minor samaras had higher amounts of soluble lipids. Dry samaras of U. laevis had higher floatability and lower air sustentation than those of U. minor. We concluded that samaras contribute to seed development by participating in carbon and water exchange. This may be especially important for U. minor, whose samaras develop before leaf emergence. The trichomes present along U. laevis samara margin may enhance water absorption and samara floatability even in turbulent waters. In general, U. minor samaras show traits that are consistent with a more drought tolerant character than U. laevis samaras, in line with the resources available both at the tree and ecosystem level for these species. Samara features may additionally reflect different adaptive strategies for seed dispersal and niche differentiation between species, by favoring hydrochory for U. laevis and anemochory for U. minor.

The importance of grasses and graminoids for sheep nutrition in Argentinean Patagonia is widely recognized. Focusing on sheep nutrition, we assessed the concentration of mineral elements in grasses growing in three ecological areas of southern Patagonia, representing a vegetation and climate gradient. With the aim of establishing potential relationships, tissue concentrations of several essential and non-essential elements for plants were determined, analyzing also soil properties. Soil and plant tissue mineral element concentrations varied between ecological areas and also depending on the particular element. The results obtained provide novel information about the nutritional characteristics of the main feeding source for sheep in southern Patagonia, but further trials will be required for improving our understanding of mineral element nutrition in relation to sheep production. EEA Santa Cruz Fil: Bahamonde, Héctor Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Universidad Nacional de la Patagonia Austral; Argentina Fil: Fernández, Victoria. Universidad Politécnica de Madrid. Escuela Técnica Superior de Ingeniería de Montes, Forestal y del Medio Natural. Departamento de Sistemas y Recursos Naturales; España Fil: Mattenet, Francisco Javier. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina Fil: Peri, Pablo Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Universidad Nacional de la Patagonia Austral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

7 Pags.- 5 Tabls.- 1 Fig., In this study we evaluated the leaf surface properties of maize populations native to different water availability environments. Leaf surface topography, wettability and gas exchange performance of five maize populations from the Sahara desert, dry (south) and humid (north-western) areas of Spain were analysed. Differences in wettability, stomatal and trichome densities, surface free energy and solubility parameter values were recorded between populations and leaf sides. Leaves from the humid Spanish population with special regard to the abaxial side, were less wettable and less susceptible to polar interactions. The higher wettability and hydrophilicity of Sahara populations with emphasis on the abaxial leaf surfaces, may favour dew deposition and foliar water absorption, hence improving water use efficiency under extremely dry conditions. Compared to the other Saharan populations, the dwarf one had a higher photosynthesis rate suggesting that dwarfism may be a strategy for improving plant tolerance to arid conditions. The results obtained for different maize populations suggest that leaf surfaces may vary in response to drought, but further studies will be required to examine the potential relationship between leaf surface properties and plant stress tolerance., This project was financed by the Spanish National Plan for Research and Development (project code AGL2013-48852-C3-1-R).

Antecedentes. La distrofia muscular cintura-cadera tipo 1B es una enfermedad con herencia autosómica dominante y secundaria a una mutación en el gen LMNA. Esta enfermedad se caracteriza por su afectación a nivel neuromuscular y cardiaco.Objetivo. Realizar diagnóstico clínico y confirmatorio molecular en una paciente con debilidad muscular proximal y sintomatología cardíaca a través de secuenciación exómica.Materiales y métodos. Se presenta el caso de una paciente de 57 años de edad con cuadro de debilidad muscular proximal progresiva principalmente en extremidades y posterior afectación cardíaca; adicionalmente, la paciente tiene múltiples familiares con la misma sintomatología. Se realizó estudio de secuenciación exómica con confirmación, por método de Sanger, de la mutación hallada y posteriormente el análisis bioinformático de esta.Resultados. La detección de la mutación R377L en el gen LMNA por secuenciación exómica con confirmación por Sanger, junto con la sintomatología clínica de la paciente y el análisis bioinformático de la mutación hallada, permitió realizar diagnóstico confirmatorio de distrofia muscular cintura-cadera tipo 1B.Conclusión. Es difícil realizar un diagnóstico clínico debido a la heterogeneidad genética del fenotipo de distrofias musculares cintura-cadera. La aproximación diagnóstica es compleja y requiere clasificar las distrofias musculares según el patrón de afectación y el patrón de herencia de la enfermedad. Adicionalmente, debido a los múltiples genes que pueden generar clínica semejante a las diferentes distrofias musculares, se recomienda realizar secuenciación exómica solicitando especial énfasis en los genes candidatos de distrofias musculares cintura-cadera. Background. Limb-girdle muscular dystrophy type 1B has a dominant autosomal inheritance pattern and is caused by a mutation in the LMNA gene. This disease has a major neuromuscular and cardiac compromise; furthermore, it belongs to the limb-girdle muscular dystrophies.Objective. To make a clinical and molecular confirmatory diagnosis in a patient with proximal muscular weakness and cardiac symptoms using whole exome sequencing.Materials and Methods. This is the case of a 57 year old patient with a slowly progressive proximal muscular weakness and cardiac compromise; furthermore, the patient has many relatives with the same clinical history. Whole exome sequencing with Sanger confirmation and bioinformatics analysis was performed on the found mutation.Results. The detection of mutation R377L in the LMNA gen by whole exome sequencing with Sanger confirmation, the bioinformatic analysis of the mutation and the symptoms exhibited by the patient allowed the confirmatory diagnosis of limb-girdle muscular dystrophy type 1b.Conclusion. Due to genetic heterogeneity in the phenotype of limb-girdle muscular dystrophies it is difficult to make a clinical diagnosis. The diagnostic approach is complex and requires classification of the muscular dystrophies according to the pattern of muscular weakness and to identify the disease inheritance pattern. Additionally, due to the multiple genes that can generate similar symptoms in the different muscular dystrophies, the authors recommend the use of whole exome sequencing with a special emphasis on the candidate genes for limb-girdle muscular dystrophies.

With this work we addressed some molecular systematic issues within the Mugil cephalus species complex. Particular attention was paid to the debated situations of: (i) Mugil liza, occurring in partial sympatry with Mugil cephalus in the northwestern Atlantic, and (ii) Mugil platanus, considered by some authors a synonymy of the former species and distributed in the southwestern Atlantic. We sequenced 79 individuals of a 465-bp portion of the mitochondrial control region (CR) from 8 western Atlantic and 2 Mediterranean localities. In addition, all CR sequences available from GenBank for the studied taxa were added to our dataset, for a total of 323 individuals. Overall, 229 haplotypes corresponding to 8 divergent monophyletic lineages were detected. Results of phylogenetic analyses were consistent with the occurrence of past speciation events producing the observed lineages. Of these lineages, 7 correspond to cryptic species and one is constituted by M. liza and M. platanus. As a matter of fact, these 2 taxa constitute a single lineage within the M. cephalus species complex. However, individuals of M. liza/M. platanus lineage analyzed by means of the 18 mitochondrial markers available in GenBank exhibited a degree of genetic diversity consistent with highly divergent populations. Of the 8 lineages detected, the Mediterraean one (type locality) corresponds to M. cephalus; the lineage M. liza/M. platanus should be named M. liza, under the priority principle, and the left 6 lineages need formal description.

The adaptive benefit of stomatal crypts remains a matter of controversy. This work studies the effect on gas exchange of cuticular rims that overarch the stomatal pore in the Mediterranean species Quercus coccifera L. growing under Mediterranean (lower relative humidities and high summer temperatures) or oceanic conditions (higher daily relative humidities and mild temperatures). After microscopic assessment of the leaf surfaces and stomatal architecture, the impact of the cuticular 'cup' on gas exchange was evaluated by employing three-dimensional finite element models. Here, we provide evidence for a high plasticity of the Q. coccifera cuticular cup, with much larger vents under oceanic conditions compared to small vents under Mediterranean conditions. This structure adds a substantial fixed resistance thereby strongly decreasing gas exchange under Mediterranean conditions. The cuticular cup, which also increases leaf internal humidity, might buffer the rapid changes in vapour pressure deficit (VPD) often observed under Mediterranean conditions. Since water loss of guard and adjacent epidermal cells regulates stomatal aperture, we suggest that this structure allows an efficient regulation of stomatal conductance and optimum use of resources under high VPD. This study provides evidence that plasticity of stomatal architecture can be an important structural component of hydraulic adaptation to different climate conditions.

A potentially fatal complication of influenza infection is the development of pneumonia, caused either directly by the influenza virus, or by secondary bacterial infection. Pneumonia related to the 2009 influenza A pandemic was found to be underestimated by commonly used pneumonia severity scores in many cases, and to be rapidly progressive, leading to respiratory failure. Confirmation of etiology by laboratory testing is warranted in such cases. Rapid antigen and immunofluorescence testing are useful screening tests, but have limited sensitivity. Confirmation of pandemic H1N1 influenza A infection can only be made by real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) or viral culture. The most effective preventive measure is annual influenza vaccination in selected individuals. Decisions to administer antiviral medications for influenza treatment or chemoprophylaxis should be based upon clinical and epidemiological factors, and should not be delayed by confirmatory laboratory testing results. Neuraminidase inhibitors (NI) are the agents of choice.

The surface of most aerial plant organs is covered with a cuticle that provides protection against multiple stress factors including dehydration. Interest on the nature of this external layer dates back to the beginning of the 19th century and since then, several studies facilitated a better understanding of cuticular chemical composition and structure. The prevailing undertanding of the cuticle as a lipidic, hydrophobic layer which is independent from the epidermal cell wall underneath stems from the concept developed by Brongniart and von Mohl during the first half of the 19th century. Such early investigations on plant cuticles attempted to link chemical composition and structure with the existing technologies, and have not been directly challenged for decades. Beginning with a historical overview about the development of cuticular studies, this review is aimed at critically assessing the information available on cuticle chemical composition and structure, considering studies performed with cuticles and isolated cuticular chemical components. The concept of the cuticle as a lipid layer independent from the cell wall is subsequently challenged, based on the existing literature, and on new findings pointing toward the cell wall nature of this layer, also providing examples of different leaf cuticle structures. Finally, the need for a re-assessment of the chemical and structural nature of the plant cuticle is highlighted, considering its cell wall nature and variability among organs, species, developmental stages, and biotic and abiotic factors during plant growth.

Natalizumab is a monoclonal antibody shown to be highly effective in the treatment of relapsing–remitting multiple sclerosis (RRMS). Patients treated with natalizumab can develop antibodies directed against this agent that may affect the efficacy and safety of the drug. In this observational study, the kinetics of the appearance and the incidence of anti-natalizumab antibodies were followed prospectively for 18 months in a cohort of 64 consecutive patients treated with natalizumab for relapsing MS. Blood samples were drawn immediately before starting natalizumab therapy and each month afterwards. The presence of antibodies against natalizumab was assessed by enzyme-linked immunosorbent assay (ELISA) in all patients. Anti-natalizumab antibodies were detected in nine (14.1%) natalizumab-treated patients, three (4.68%) of whom were transiently positive while six (9.37%) were persistently positive (these patients discontinued natalizumab). All positive titres were observed during the first 4 months of treatment. One patient with a hypersensitivity reaction also had persistent antibodies. We conclude that antibodies against natalizumab develop early, within the first 6 months of therapy with natalizumab. Although no antibodies were detected after 4 months of therapy in this particular study, this does not rule out their development later on in exceptional cases.

Alzheimer disease (AD), Frontotemporal lobar degeneration (FTD), Amyotrophic lateral sclerosis (ALS) and Parkinson disease (PD) have a certain degree of clinical, pathological and molecular overlap. Previous studies indicate that causative mutations in AD and FTD/ALS genes can be found in clinical familial AD. We examined the presence of causative and low frequency coding variants in the AD, FTD, ALS and PD Mendelian genes, in over 450 families with clinical history of AD and over 11,710 sporadic cases and cognitive normal participants from North America. Known pathogenic mutations were found in 1.05% of the sporadic cases, in 0.69% of the cognitively normal participants and in 4.22% of the families. A trend towards enrichment, albeit non-significant, was observed for most AD, FTD and PD genes. Only PSEN1 and PINK1 showed consistent association with AD cases when we used ExAC as the control population. These results suggest that current study designs may contain heterogeneity and contamination of the control population, and that current statistical methods for the discovery of novel genes with real pathogenic variants in complex late onset diseases may be inadequate or underpowered to identify genes carrying pathogenic mutations., Author summary In this study we provide further genetic evidence of the clinical, pathological and molecular overlap between neurodegenerative diseases. We screened the known Mendelian genes in Alzheimer disease (AD), Frontotemporal Dementia (FTD), Parkinson disease (PD) and Amyotrophic Lateral Sclerosis (ALS) disease in over 13,292 individuals. We report the presence of known pathogenic mutations in AD, FTD and PD genes in both sporadic and familial late onset AD cases and even cognitively normal individuals, pointing to contamination of control cohorts with asymptomatic cases. We also observed an enrichment, although not statistically significant, in most of AD, FTD and PD genes, reinforcing the idea of pathologic crossover among these diseases. However, the fact that genes carrying known pathogenic mutation do not show significant association calls for a reevaluation of current statistical methods.

BACKGROUND: A strategy to supply Ca directly to fruits as a tool for improving peach quality has been devised and tested under field conditions. Since peaches in the area of study (Calanda, Spain) are routinely bagged shortly after thinning, a method based on the application of Ca gels to the fruit surface was introduced. The effect of surface treatments was assessed in terms of quality, nutrient balance and surface deposition. RESULTS: Application of Ca-containing formulations increased mesocarp and exocarp Ca concentrations, providing evidence for the penetration of Ca through the peach skin. Surface Ca treatments had a particular mode of deposition and in some instances improved the shelf life of fruits without affecting their quality. CONCLUSION: Surface treatment with Ca-containing gels appears to be a viable approach to increase fruit Ca, quality and storability of bagged peach cultivars and should be optimised in future studies. Copyright © 2009 Society of Chemical Industry

The final version is available at: http://www.springerlink.com/content/y62g0r6712184032/fulltext.pdf, Plants grown in calcareous, high pH soils develop Fe deficiency chlorosis. While the physiological parameters of Fe-deficient leaves have been often investigated, there is a lack of information regarding structural leaf changes associated with such abiotic stress. Iron-sufficient and Fe-deficient pear and peach leaves have been studied, and differences concerning leaf epidermal and internal structure were found. Iron deficiency caused differences in the aspect of the leaf surface, which appeared less smooth in Fe-deficient than in Fe-sufficient leaves. Iron deficiency reduced the amount of soluble cuticular lipids in peach leaves, whereas it reduced the weight of the abaxial cuticle in pear leaves. In both plant species, epidermal cells were enlarged as compared to healthy leaves, whereas the size of guard cells was reduced. In chlorotic leaves, bundle sheaths were enlarged and appeared disorganized, while the mesophyll was more compacted and less porous than in green leaves. In contrast to healthy leaves, chlorotic leaves of both species showed a significant transient opening of stomata after leaf abscission (Iwanoff effect), which can be ascribed to changes found in epidermal and guard cells. Results indicate that Fe-deficiency may alter the barrier properties of the leaf surface, which can significantly affect leaf water relations, solute permeability and pest and disease resistance., This study was supported by the Spanish Ministry of Science and Education (MEC, grants AGL2006-01416 and AGL2007-61948, co-financed with FEDER), the European Commission (ISAFRUIT project, Thematic Priority 5-Food Quality and Safety of the 6th Framework Programme of RTD; Contract no. FP6-FOOD-CT-2006-016279) and the Aragón Government (group A03). V.F. was supported by a “Juan de la Cierva”-MEC post-doctoral contract, co-financed by the European Social Fund. T.E. was supported by the CAI Europa XXI for a short term stay at the EEAD-CSIC.

This work is licensed under a Creative Commons Attribution 4.0 International License.-- et al., Cardiac angiosarcoma (CAS) is a rare malignant tumour whose genetic basis is unknown. Here we show, by whole-exome sequencing of a TP53-negative Li-Fraumeni-like (LFL) family including CAS cases, that a missense variant (p.R117C) in POT1 (protection of telomeres 1) gene is responsible for CAS. The same gene alteration is found in two other LFL families with CAS, supporting the causal effect of the identified mutation. We extend the analysis to TP53-negative LFL families with no CAS and find the same mutation in a breast AS family. The mutation is recently found once in 121,324 studied alleles in ExAC server but it is not described in any other database or found in 1,520 Spanish controls. In silico structural analysis suggests how the mutation disrupts POT1 structure. Functional and in vitro studies demonstrate that carriers of the mutation show reduced telomere-bound POT1 levels, abnormally long telomeres and increased telomere fragility., We are grateful to T. de Lange (The Rockefeller University) and K. Collins (The University of California) for providing POT1 and TPP1 plasmids, respectively. J.B.’s laboratory is partially funded by the Spanish Ministry of Health PI12/00070, the Spanish Ministry of Science and Innovation (INNPRONTA 2012) and the Spanish Research Network on Rare diseases (CIBERER). O.C. is granted by the CIBERER and C.B.-B. by the PI12/00070 supported by FEDER funds. P.G.-P. is partially supported by the Spanish Ministry of Health PI11/0699, PI12/01941 and RD12/0042/0066. M.A.B.’s laboratory is funded with the Spanish Ministry of Science and Innovation, projects SAF2008-05384 and 2007-A-200950 (TELOMARKER), European Research Council Advanced grant GA#232854, the Körber Foundation, Fundación Botín and Fundación Lilly. R.P.’ lab is partially funded by PI11/0949 Supported by FEDER funds.

p73 shares structural and functional homology to p53. p73 generates different proteins using alternative promoters and splicing which have different biological characteristics. We investigated the pattern (monoallelic or biallelic) of expression of TAp73 and DeltaNp73 in normal lymphocytes and lymphomas using two p73 polymorphisms. We found monoallelic expression of TAp73 in normal lymphocytes and tumors, and a selective expression of AT allele in all cases. Moreover, the quantitative expression analysis revealed DeltaNp73 over-expression in both B- and T-cell lymphomas comparing with normal lymphoid cells, suggesting a role in tumorigenesis. Finally, we have confirmed that although DeltaNp73 over-expression could be an alternative mechanism of p53 inactivation, both alterations may appear together.

Gastric neuroendocrine tumours (NETs) arise from enterochromaffin-like cells, which are located in oxyntic glands within the stomach. Type I tumours represent 70-80% of gastric NETs and are associated with hypergastrinaemia, chronic atrophic gastritis and achlorhydria. Gastrin is involved in the endocrine regulation of gastric acid production. Most type I gastric NETs are sporadic, have a good prognosis and their genetic basis are unknown. We performed an exome sequencing study in a family with consanguineous parents and 10 children, five of whom were affected by type I gastric NET. Atypical clinical traits included an earlier age of onset (around 30 years), aggressiveness (three had nodal infiltration requiring total gastrectomy and one an adenocarcinoma) and iron-deficiency rather than megaloblastic anaemia. We identified a homozygous missense mutation in the 14th exon of the ATP4A gene (c.2107C>T), which encodes the proton pump responsible for acid secretion by gastric parietal cells. The amino acid p.Arg703Cys is highly conserved across species and originates a change of one of the transmembrane domains that avoids the liberation of protons from cells to stomach. This is consistent with the achlorhydria that was observed in the affected individuals. No germline or somatic mutations in the ATP4A gene were found in sporadic gastric NET patients. Based on the results of this large family, it seems that this atypical form of gastric NET has an earlier age of onset, behaves more aggressively and has atypical clinical traits that differentiated from other studied cases.

Application of foliar iron (Fe) sprays is a common means of correcting Fe deficiency of agricultural crops. However, variable plant responses to iron sprays, ranging from no effect to defoliation, have often been described in the Fe-fertilization literature. Knowledge is still limited concerning the mechanisms of penetration of a leaf-applied, Fe-containing solution and the role of Fe in the leaf. The complex and multi-disciplinary character of the factors determining the effects of Fe sprays hinder the development of suitable foliar fertilization strategies, applicable under variable local conditions and for different plant types. This review describes some key factors involved on the process of penetration of a leaf-applied, Fe-containing solution before briefly analyzing the available foliar Fe-fertilization literature. Iron chemistry, leaf penetration, and plant-nutrition principles will be merged with the aim of clarifying the constraints, opportunities, and future perspectives of foliar Fe sprays to cure plant Fe deficiency.

Experiments were conducted to assess the distribution of foliar applied Fe-containing compounds using microbial siderophores. Fe was measured in leaf fluid obtained by centrifugation according to a determination method based on Fe chelation by desferrioxamine E and HPLC separation on a reversed phase column. To avoid sample Fe contamination, treatments were only applied to a part of the leaf following a systematic and reproducible procedure and iron concentration was exclusively determined in fluid obtained from non-treated leaf surfaces. The increase in leaf fluid Fe concentration associated with the distribution of leaf applied Fe-siderophores, Fe–EDTA and FeSO4 × 7H2O was evaluated using Vicia faba L., Nicotiana tabacum L. and Citrus madurensis Lour. plants. The method proved useful to investigate the process of leaf Fe penetration and its distribution within the plant. Evidence of the penetration and distribution of leaf applied Fe-rhizoferrin, Fe-coprogen hydrolysis products and Fe-dimerum acid is presented in this study.

Experiments to assess the re-greening and the distribution of leaf-applied iron (Fe) within the aerial organs were developed with tobacco (Nicotiana tabacum L.) plants. Fe (III)-citrate and Fe-dimerum acid were applied to a part of the leaf and plant re-greening was monitored for 6 weeks after treatment. Fluid Fe increments associated with foliar Fe application were measured within 3 days after application. Fe distribution from the site of application within the leaf and the untreated above-ground plant parts was evaluated. Leaf fluid Fe concentration was determined by a novel procedure based on Fe chelation by desferrioxamine E and high performance liquid chromatography separation on a reversed-phase column. The ferrioxamine E method enabled accurate determination of small amounts of Fe present in leaf fluid. Foliar Fe-dimerum acid and Fe (III)-citrate treatment to chlorotic tobacco plants induced new growth re-greening. It was concluded that the applied Fe-containing compounds penetrated the leaf, were distributed within the plant and that Fe supplied as both complexes could be utilized by the plant cell.

The human leukocyte antigen (HLA) class II DR2 haplotype (DRB1*1501, DQA1*0102, DQB1*0602) has been associated with multiple sclerosis (MS) in all ethnic groups and very strongly in Caucasians. To investigate the possible HLA class II (DRB1, DQA1 and DQB1) associations with MS in Malaga, southern Spain. We analysed the HLA class II sub-regions DRB1, DQA1 and DQB1 by polymerase chain reaction (PCR) and sequence-specific oligonucleotide probe hybridization (PCR/SSO) for DRB1 and DQB1 and with sequence-specific primers (PCR/SSP) for DRB1 subtypes and DQA1. Possible HLA class II associations with clinical MS characteristics were investigated in 149 subjects with and 160 without MS. Associations were detected between MS and the HLA class II alleles DRB1*1501 (45.6 % vs. 21.3%, p = 0.001), DQA1*0102 (44% vs. 29.4%, p = 0.001) and DQB1*0602 (45% vs. 20.6%, p = 0.001). The DR2 haplotype (DRB1*1501, DQA1*0102, DQB1*0602) was associated with MS (43.6 % vs. 20%, p = 0.002). DQB1*0602 was the only allele that maintained an association with MS in a logistic regression model. No HLA class II alleles or genotypes were significantly associated with any clinical characteristics of MS. Our results confirm the positive association of the DR2 haplotype with MS, particularly the allele DQB1*0602, in the population studied. DR4 was not associated with the disease in Malaga. HLA class II alleles or haplotypes were not associated with clinical or demographic characteristics, or clinical form or severity of MS.

The plant cuticle has traditionally been conceived as an independent hydrophobic layer that covers the external epidermal cell wall. Due to its complexity, the existing relationship between cuticle chemical composition and ultra-structure remains unclear to date. This study aimed to examine the link between chemical composition and structure of isolated, adaxial leaf cuticles of Eucalyptus camaldulensis and E. globulus by the gradual extraction and identification of lipid constituents (cutin and soluble lipids), coupled to spectroscopic and microscopic analyses. The soluble compounds and cutin monomers identified could not be assigned to a concrete internal cuticle ultra-structure. After cutin depolymerization, a cellulose network resembling the cell wall was observed, with different structural patterns in the regions ascribed to the cuticle proper and cuticular layer, respectively. Our results suggest that the current cuticle model should be revised, stressing the presence and major role of cell wall polysaccharides. It is concluded that the cuticle may be interpreted as a modified cell wall region which contains additional lipids. The major heterogeneity of the plant cuticle makes it difficult to establish a direct link between cuticle chemistry and structure with the existing methodologies.

While plant roots are specialized organs for the uptake and transport of water and nutrients, the absorption of gaseous or liquid mineral elements by aerial plant parts has been recognized since more than one century. Nitrogen (N) is an essential macronutrient which generally absorbed either as nitrate (NO(-) 3) or ammonium (NH(+) 4) by plant roots. Gaseous nitrogen pollutants like N dioxide (NO2) can also be absorbed by plant surfaces and assimilated via the NO(-) 3 assimilation pathway. The subsequent NO(-) 3 flux may induce or repress the expression of various NO(-) 3-responsive genes encoding for instance, the transmembrane transporters, NO(-) 3/NO(-) 2 (nitrite) reductase, or assimilatory enzymes involved in N metabolism. Based on the existing information, the aim of this review was to theoretically analyze the potential link between foliar NO2 absorption and N transport and metabolism. For such purpose, an overview of the state of knowledge on the NO(-) 3 transporter genes identified in leaves or shoots of various species and their roles for NO(-) 3 transport across the tonoplast and plasma membrane, in addition to the process of phloem loading is briefly provided. It is assumed that a NO2-induced accumulation of NO(-) 3/NO(-) 2 may alter the expression of such genes, hence linking transmembrane NO(-) 3 transporters and foliar uptake of NO2. It is likely that NRT1/NRT2 gene expression and species-dependent apoplastic buffer capacity may be also related to the species-specific foliar NO2 uptake process. It is concluded that further work focusing on the expression of NRT1 (NRT1.1, NRT1.7, NRT1.11, and NRT1.12), NRT2 (NRT2.1, NRT2.4, and NRT2.5) and chloride channel family genes (CLCa and CLCd) may help us elucidate the physiological and metabolic response of plants fumigated with NO2.

The calcifuge and calcicole character of wild plants has been related to nutrient availability shortages, including iron (Fe)-deficiency. Surprisingly, just a few studies examined the relation between root Fe uptake and plant distribution in different soil types. We assessed the root Fe acquisition efficiency of two Ulmus species with calcareous (U. minor) and siliceous (U. laevis) soil distribution patterns in the Iberian Peninsula. Seedlings of both elm species were grown hydroponically with different Fe concentrations during six weeks. Plant physiological responses to Fe-limiting conditions were evaluated as were the ferric reductase activity and proton (H+) extrusion capacity of the roots. Iron deprived elm seedlings of both species were stunted and suffered severe Fe-chlorosis symptoms. After Fe re-supply leaf chlorophyll concentrations rose according to species-dependent patterns. While U. minor leaves and seedlings re-greened evenly, U. laevis did so along the nerves of new growing leaves. Ulmus minor had a higher root ferric reductase activity and H+-extrusion capability than U. laevis and maintained a better nutrient balance when grown under Fe-limiting conditions. The two elm species were found to have different iron acquisition efficiencies which may be related to their natural distribution in calcareous and siliceous soils of the Iberian Peninsula.

The presence and characteristics of cuticle polysaccharides have been demonstrated by staining and spectroscopic methods, but their location in the cuticle remains unclear. Furthermore, according to the prevailing model, polysaccharides are believed to be restricted to the cuticular layer and absent in the cuticle proper. With the aim of gaining insight into cuticular ultra-structure focussing on polysaccharides, cellulose and pectins have been identified and located in the transversal sections of isolated and intact adaxial leaf cuticles of Eucalyptus globulus, Populus × canescens and Pyrus communis by means of enzyme gold-labelling (Au-cellulase, EC 3.2.1.4, and -pectinase, EC 3.2.1.15) and transmission electron microscopy (TEM). The structure of the interface between the cuticle and the cell wall underneath was observed to influence the process of enzymatic isolation of leaf cuticles. Cellulose and pectins were detected for the first time in enzymatically isolated cuticles, sometimes appearing closely underneath the epicuticular wax layer. The location and presence of polysaccharides in intact and isolated leaf cuticles may have multiple implications, such as when estimating the bi-directional transport of substances between plant organs and the surrounding environment, or when interpreting organ ontogeny. The results are discussed within a plant ontological and ecophysiological context.

Little is known on the participation of immunoglobulin isotypes and subclasses in the pathogenesis of the severe disease caused by the pandemic influenza virus (influenza A(H1N1)pdm09).(1) To evaluate the association between plasma levels of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE and outcome in patients with severe pandemic influenza. (2) To evaluate the association between immunoglobulin and cytokine levels in these patients.40 critically ill patients with community acquired pneumonia and influenza A(H1N1)pdm09 infection were recruited from November 2010 to February 2011. Plasma samples were collected during the first 24h following admission to the ICU. Immunoglobulins and 17 major cytokines were profiled in plasma.15 patients died (37.5%). When the association between clinical variables and prognosis was assessed, prior immunosuppression, APACHE II score, levels of IgG2 and levels of IgM were associated with outcome in a univariate Cox regression analysis. Kaplan Meier analysis showed that patients with levels of IgG2 and IgM59 and58 mg/dl respectively died earlier. Multivariate Cox regression analysis showed that APACHE II score and levels of IgM were the best predictors of outcome, being levels of IgM a protective factor against mortality. IgM was the immunoglobulin showing the largest number of negative correlations with cytokine levels.Our results support a central role of IgM in preventing uncontrolled inflammatory response and mortality in severe pandemic influenza. Early assessment of IgM could contribute to guide clinical decisions in these patients.

The application of agrochemical sprays to the aerial parts of crop plants is an important agricultural practice world-wide. While variable effectiveness is often seen in response to foliar treatments, there is abundant evidence showing the beneficial effect of foliar fertilizers in terms of improving the metabolism, quality, and yields of crops. This mini-review is focused on the major bottlenecks associated with the uptake and translocation of foliar-applied nutrient solutions. A better understanding of the complex scenario surrounding the ultimate delivery of foliar-applied nutrients to sink cells and organs is essential for improving the effectiveness and performance of foliar fertilizers.

Background Gene expression profiling (GEP) in cells obtained from peripheral blood has shown that this is a very useful approach for biomarker discovery and for studying molecular pathogenesis of prevalent diseases. While there is limited literature available on gene expression markers associated with Chronic Obstructive Pulmonary Disease (COPD), the transcriptomic picture associated with critical respiratory illness in this disease is not known at the present moment. Findings By using Agilent microarray chips, we have profiled gene expression signatures in the whole blood of 28 COPD patients hospitalized with different degrees of respiratory compromise.12 of them needed of admission to the ICU, whilst 16 were admitted to the Respiratory Medicine Service. GeneSpring GX 11.0 software was used for performing statistical comparisons of transcript levels between ICU and non-ICU patients. Ingenuity pathway analysis 8.5 (IPA) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate and visualize genes by function and pathway (gene ontology). T-test showed evidence of 1501 genes differentially expressed between ICU and non-ICU patients. IPA and KEGG analysis of the most representative biological functions revealed that ICU patients had increased levels of neutrophil gene transcripts, being [cathepsin G (CTSG)], [elastase, neutrophil expressed (ELANE)], [proteinase 3 (PRTN3)], [myeloperoxidase (MPO)], [cathepsin D (CTSD)], [defensin, alpha 3, neutrophil-specific (DEFA3)], azurocidin 1 (AZU1)], and [bactericidal/permeability-increasing protein (BPI)] the most representative ones. Proteins codified by these genes form part of the azurophilic granules of neutrophils and are involved in both antimicrobial defence and tissue damage. This “neutrophil signature” was paralleled by the necessity of advanced respiratory and vital support, and the presence of bacterial infection. Conclusion Study of transcriptomic signatures in blood suggests an essential role of neutrophil proteases in COPD patients with critical respiratory illness. Measurement and modulation of the expression of these genes could present an option for clinical monitoring and treatment of severe COPD exacerbations.