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1.
Chemosphere ; 356: 141887, 2024 May.
Article in English | MEDLINE | ID: mdl-38583530

ABSTRACT

Microplastics pose risks to marine organisms through ingestion, entanglement, and as carriers of toxic additives and environmental pollutants. Plastic pre-production pellet leachates have been shown to affect the development of sea urchins and, to some extent, mussels. The extent of those developmental effects on other animal phyla remains unknown. Here, we test the toxicity of environmental mixed nurdle samples and new PVC pellets for the embryonic development or asexual reproduction by regeneration of animals from all the major animal superphyla (Lophotrochozoa, Ecdysozoa, Deuterostomia and Cnidaria). Our results show diverse, concentration-dependent impacts in all the species sampled for new pellets, and for molluscs and deuterostomes for environmental samples. Embryo axial formation, cell specification and, specially, morphogenesis seem to be the main processes affected by plastic leachate exposure. Our study serves as a proof of principle for the potentially catastrophic effects that increasing plastic concentrations in the oceans and other ecosystems can have across animal populations from all major animal superphyla.


Subject(s)
Invertebrates , Microplastics , Plastics , Water Pollutants, Chemical , Animals , Water Pollutants, Chemical/toxicity , Water Pollutants, Chemical/analysis , Plastics/toxicity , Invertebrates/drug effects , Microplastics/toxicity , Embryonic Development/drug effects
2.
Nat Commun ; 14(1): 3970, 2023 07 05.
Article in English | MEDLINE | ID: mdl-37407549

ABSTRACT

During early development of the sea urchin embryo, activation of ERK signalling in mesodermal precursors is not triggered by extracellular RTK ligands but by a cell-autonomous, RAS-independent mechanism that was not understood. We discovered that in these cells, ERK signalling is activated through the transcriptional activation of a gene encoding a protein related to Kinase Suppressor of Ras, that we named KSR3. KSR3 belongs to a family of catalytically inactive allosteric activators of RAF. Phylogenetic analysis revealed that genes encoding kinase defective KSR3 proteins are present in most non-chordate metazoa but have been lost in flies and nematodes. We show that the structure of KSR3 factors resembles that of several oncogenic human RAF mutants and that KSR3 from echinoderms, cnidarians and hemichordates activate ERK signalling independently of RAS when overexpressed in cultured cells. Finally, we used the sequence of KSR3 factors to identify activating mutations of human B-RAF. These findings reveal key functions for this family of factors as activators of RAF in RAS-independent ERK signalling in invertebrates. They have implications on the evolution of the ERK signalling pathway and suggest a mechanism for its co-option in the course of evolution.


Subject(s)
MAP Kinase Signaling System , Signal Transduction , Animals , Humans , Phylogeny , MAP Kinase Signaling System/genetics , Protein Kinases/metabolism , Proto-Oncogene Proteins c-raf/metabolism
3.
Int J Mol Sci ; 24(6)2023 Mar 13.
Article in English | MEDLINE | ID: mdl-36982516

ABSTRACT

Selenium (Se) is an essential micronutrient of fundamental importance to human health and the main Se source is from plant-derived foods. Plants mainly take up Se as selenate (SeO42-), through the root sulfate transport system, because of their chemical similarity. The aims of this study were (1) to characterize the interaction between Se and S during the root uptake process, by measuring the expression of genes coding for high-affinity sulfate transporters and (2) to explore the possibility of increasing plant capability to take up Se by modulating S availability in the growth medium. We selected different tetraploid wheat genotypes as model plants, including a modern genotype, Svevo (Triticum turgidum ssp. durum), and three ancient Khorasan wheats, Kamut, Turanicum 21, and Etrusco (Triticum turgidum ssp. turanicum). The plants were cultivated hydroponically for 20 days in the presence of two sulfate levels, adequate (S = 1.2 mM) and limiting (L = 0.06 mM), and three selenate levels (0, 10, 50 µM). Our findings clearly showed the differential expression of genes encoding the two high-affinity transporters (TdSultr1.1 and TdSultr1.3), which are involved in the primary uptake of sulfate from the rhizosphere. Interestingly, Se accumulation in shoots was higher when S was limited in the nutrient solution.


Subject(s)
Selenium , Triticum , Humans , Selenic Acid , Triticum/metabolism , Tetraploidy , Sulfates/metabolism , Selenium/metabolism , Genotype
4.
J Clin Med ; 11(12)2022 Jun 11.
Article in English | MEDLINE | ID: mdl-35743428

ABSTRACT

Anticholinergic burden (AB) is related to cognitive impairment (CI) and older complex chronic patients (OCCP) are more susceptible. Our objective was to evaluate the predictive value of ten anticholinergic scales to predict a potential CI due to anticholinergic pharmacotherapy in OCCP. An eight-month longitudinal multicentre study was carried out in a cohort of OCCP, in treatment with at least one anticholinergic drug and whose cognition status had been evaluated by Pfeiffer test twice for a period of 6-15 months. CI was considered when the Pfeiffer test increased 2 or more points. AB was detected using ten scales included on the Anticholinergic Burden Calculator. An ROC curve analysis was performed to assess the discriminative capacity of the scales to predict a potential CI and the cut-off point of AB that obtains better validity indicators. 415 patients were included (60.2% female, median age of 85 years (IQR = 11)). 190 patients (45.8%) manifested CI. Only the DBI (Drug Burden Index) showed statistically significant differences in the median AB between patients without CI and with CI (0.5 (1.00) vs. 0.67 (0.65), p = 0.006). At the ROC curve analysis, statistically significant values were obtained only with the DBI (AUC: 0.578 (0.523-0.633), p = 0.006). The cut-off point with the greatest validity selected for the DBI was an AB of 0.41 (moderate risk) (sensitivity = 81%, specificity = 36%, PPV = 51%). The DBI is the scale with the greatest discriminatory power to detect OCCP at risk of CI and the best cut-off point is a load value of 0.41.

5.
Theor Appl Genet ; 134(12): 4013-4024, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34477900

ABSTRACT

KEY MESSAGE: The suppression of the HYD-1 gene by a TILLING approach increases the amount of ß-carotene in durum wheat kernel. Vitamin A deficiency is a major public health problem that affects numerous countries in the world. As humans are not able to synthesize vitamin A, it must be daily assimilated along with other micro- and macronutrients through the diet. Durum wheat is an important crop for Mediterranean countries and provides a discrete amount of nutrients, such as carbohydrates and proteins, but it is deficient in some essential micronutrients, including provitamin A. In the present work, a targeting induced local lesions in genomes strategy has been undertaken to obtain durum wheat genotypes biofortified in provitamin A. In detail, we focused on the suppression of the ß-carotene hydroxylase 1 (HYD1) genes, encoding enzymes involved in the redirection of ß-carotene toward the synthesis of the downstream xanthophylls (neoxanthin, violaxanthin and zeaxanthin). Expression analysis of genes involved in carotenoid biosynthesis revealed a reduction of the abundance of HYD1 transcripts greater than 50% in mutant grain compared to the control. The biochemical profiling of carotenoid in the wheat mutant genotypes highlighted a significant increase of more than 70% of ß-carotene compared to the wild-type sibling lines, with no change in lutein, α-carotene and zeaxanthin content. This study sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat and provides new genotypes that represent a good genetic resource for future breeding programs focused on the provitamin A biofortification through non-transgenic approaches.


Subject(s)
Metabolic Engineering , Mixed Function Oxygenases/genetics , Provitamins/biosynthesis , Seeds/chemistry , Triticum/genetics , Vitamin A/biosynthesis , Carotenoids , Edible Grain/chemistry , Edible Grain/genetics , Food, Fortified , Gene Knockout Techniques , Genotype , Phylogeny , Plant Breeding , Triticum/chemistry , Xanthophylls , Zeaxanthins/biosynthesis
6.
Development ; 148(2)2021 01 20.
Article in English | MEDLINE | ID: mdl-33298464

ABSTRACT

During sea urchin development, secretion of Nodal and BMP2/4 ligands and their antagonists Lefty and Chordin from a ventral organiser region specifies the ventral and dorsal territories. This process relies on a complex interplay between the Nodal and BMP pathways through numerous regulatory circuits. To decipher the interplay between these pathways, we used a combination of treatments with recombinant Nodal and BMP2/4 proteins and a computational modelling approach. We assembled a logical model focusing on cell responses to signalling inputs along the dorsal-ventral axis, which was extended to cover ligand diffusion and enable multicellular simulations. Our model simulations accurately recapitulate gene expression in wild-type embryos, accounting for the specification of ventral ectoderm, ciliary band and dorsal ectoderm. Our model simulations further recapitulate various morphant phenotypes, reveal a dominance of the BMP pathway over the Nodal pathway and stress the crucial impact of the rate of Smad activation in dorsal-ventral patterning. These results emphasise the key role of the mutual antagonism between the Nodal and BMP2/4 pathways in driving early dorsal-ventral patterning of the sea urchin embryo.


Subject(s)
Body Patterning , Embryo, Nonmammalian/metabolism , Paracentrotus/embryology , Signal Transduction , Transforming Growth Factor beta/metabolism , Animals , Blastula/metabolism , Body Patterning/drug effects , Body Patterning/genetics , Bone Morphogenetic Proteins/metabolism , Cell Lineage/drug effects , Cell Lineage/genetics , Computer Simulation , Embryo, Nonmammalian/drug effects , Gene Expression Regulation, Developmental/drug effects , Glycoproteins/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Models, Biological , Morpholinos/pharmacology , Nodal Protein/metabolism , Paracentrotus/drug effects , Paracentrotus/genetics , Phenotype , Probability , Signal Transduction/drug effects , Signal Transduction/genetics , Stochastic Processes
7.
Curr Top Dev Biol ; 140: 283-316, 2020.
Article in English | MEDLINE | ID: mdl-32591077

ABSTRACT

Specification of the main axes of polarity of the embryo is an essential process during embryonic development. In many species, this process is achieved by the localization of maternal factors into discrete regions of the egg. However, in other animals, like in amniotes and in echinoderms, the considerable plasticity of the early blastomeres seems to preclude the existence of maternal determinants and the mechanisms by which the radial symmetry of the egg is broken remain largely enigmatic. In this chapter, we review recent progress on the identification of maternal components involved in symmetry breaking and dorsal-ventral (D/V) axis formation of the sea urchin embryo. We will first review some key experiments on D/V axis formation from classical embryologists that provided evidence for a weak maternal D/V prepattern. We will then detail more recent molecular analyses that established the critical role played by Nodal signaling in allocating cell fates along the secondary axis and led to the discovery that maternal transcription factors such as the Sry-related HMG box B1 (SoxB1), the Octamer binding factor1/2 (Oct1/2), the T-cell factor/Lymphoid enhancer-binding factor (TCF/LEF) and the Erythroblastosis virus E26 Oncogene Homolog (ETS) domain transcriptional repressor Translocation-Ets-Leukemia virus protein (Yan/Tel) as well as maternal signaling molecules like Univin are essential for the initiation of nodal expression. Finally, we will describe recent advances that uncovered a role in symmetry breaking and dorsal-ventral axis orientation for the transforming growth factor beta (TGF-beta)-like factor Panda, which appears to be both necessary and sufficient for D/V axis orientation. Therefore, even in the highly regulative sea urchin embryo, the activity of localized maternal factors provides the embryo with a blueprint of the D/V axis.


Subject(s)
Blastomeres/metabolism , Body Patterning/genetics , Embryo, Nonmammalian/metabolism , Gene Expression Regulation, Developmental , Sea Urchins/genetics , Animals , Blastomeres/cytology , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/embryology , Maternal Inheritance/genetics , Models, Genetic , Nodal Protein/genetics , Nodal Protein/metabolism , Sea Urchins/embryology , Signal Transduction/genetics
8.
Afr J Reprod Health ; 24(1): 81-86, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32358939

ABSTRACT

Menstrual disorders are highly prevalent and generate discomfort, anxiety, and more gynecological visits. There has been an increase in the practice of physical exercise among the general population. This study determined the association between physical exercise and menstrual disorders. An observational analytical study was conducted during 2016 in women of reproductive age from southeast Spain. Information was collected on sociodemographic, gynecological, and nutritional status variables, and the International Physical Activity Questionnaire (IPAQ) was used to gather data on physical exercise. Quantitative/qualitative variables were compared using the non- parametric Mann-Whitney U test and categorical variables with the chi-square (χ2) or Fisher's exact test, followed by multivariate logistic regression analysis. The study included 122 women; physical exercise was practiced by 50%; menstrual disorders were reported by 53.28 %. The practice of physical exercise, h/week of physical exercise, body type, level of physical exercise, and marital status were associated with the presence of menstrual cycle disorders (p˂0.05). This association persisted (p=0.042) after adjustment of the analysis for impaired nutritional status, medication consumption, the presence of disease, age, marital status, age at menarche, and number of children. A high physical activity not related to the practice of sport is associated with menstrual disorders.


Subject(s)
Exercise/physiology , Menstrual Cycle/physiology , Menstruation Disturbances/epidemiology , Adolescent , Adult , Female , Humans , Leisure Activities , Menstruation Disturbances/etiology , Prevalence , Socioeconomic Factors , Surveys and Questionnaires
9.
Int J Mol Sci ; 20(22)2019 Nov 14.
Article in English | MEDLINE | ID: mdl-31739436

ABSTRACT

Macro- and micronutrients, essential for the maintenance of human metabolism, are assimilated daily through the diet. Wheat and other major cereals are a good source of nutrients, such as carbohydrates and proteins, but cannot supply a sufficient amount of essential micronutrients, including provitamin A. As vitamin A deficiency (VAD) leads to several serious diseases throughout the world, the biofortification of a major staple crop, such as wheat, represents an effective way to preserve human health in developing countries. In the present work, a key enzyme involved in the branch of carotenoids pathway producing ß-carotene, lycopene epsilon cyclase, has been targeted by a Targeting Induced Local Lesions in Genomes (TILLING) approach in a "block strategy" perspective. The null mutant genotype showed a strong reduction in the expression of the lcyE gene and also interesting pleiotropic effects on an enzyme (ß-ring hydroxylase) acting downstream in the pathway. Biochemical profiling of carotenoids in the wheat mutant lines showed an increase of roughly 75% in ß-carotene in the grains of the complete mutant line compared with the control. In conclusion, we describe here the production and characterization of a new wheat line biofortified with provitamin A obtained through a nontransgenic approach, which also sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat.


Subject(s)
Biofortification , Genetic Engineering , Triticum/genetics , Triticum/metabolism , Vitamin A/metabolism , Alleles , Base Sequence , Carotenoids/metabolism , Gene Expression Regulation, Plant , Gene Targeting , Genetic Engineering/methods , Genomics/methods , Humans , Intramolecular Lyases/genetics , Intramolecular Lyases/metabolism , Mutation , Phylogeny , Plants, Genetically Modified
10.
Methods Cell Biol ; 151: 239-282, 2019.
Article in English | MEDLINE | ID: mdl-30948011

ABSTRACT

With the completion of the genome sequencing projects, a new challenge for developmental biologists is to assign a function to the thousands of genes identified. Expression of exogenous mRNAs is a powerful, versatile and rapid technique that can be used to study gene function during development of the sea urchin. This chapter describes how this technique can be used to analyze gene function in echinoderm embryos, how it can be combined with cell transplantation to perform mosaic analysis and how it can be applied to identify downstream targets genes of transcription factors and signaling pathways. We describe specific examples of the use of overexpression of mRNA to analyze gene function, mention the benefits and current limitations of the technique and emphasize the importance of using different controls to assess the specificity of the effects observed. Finally, this chapter details the different steps, vectors and protocols for in vitro production of mRNA and phenotypic analysis.


Subject(s)
Gene Expression Regulation, Developmental/genetics , Genomics/methods , RNA, Messenger/genetics , Sea Urchins/genetics , Animals , Embryo, Nonmammalian , Gene Regulatory Networks/genetics , Sea Urchins/growth & development , Signal Transduction/genetics , Transcription Factors/genetics
11.
Nutrients ; 11(3)2019 Feb 26.
Article in English | MEDLINE | ID: mdl-30813572

ABSTRACT

Gluten proteins, major determinants of the bread-making quality of wheat, are related to several digestive disorders. Advances in plant genetic breeding have allowed the production of wheat lines with very low gliadin content through the use of RNAi and gene editing technologies. In this review, we carried out a comprehensive study of the application of these cutting-edge technologies towards the development of wheat lines devoid of immunogenic gluten, and their genetic, nutritional and clinical characterization. One line, named E82, showed outstanding nutritional properties, with very low immunogenic gluten and a low stimulation capacity of T-cells from celiac patients. Moreover, a clinical trial with non-celiac wheat sensitivity (NCWS) patients showed that the consumption of bread made with this E82 low gliadin line induced positive changes in the gut microbiota composition.


Subject(s)
Glutens/metabolism , Triticum/genetics , Triticum/metabolism , Diet, Gluten-Free , Glutens/chemistry , Humans , Plants, Genetically Modified , Triticum/chemistry
13.
Nefrología (Madrid) ; 38(4): 394-400, jul.-ago. 2018. tab, graf
Article in Spanish | IBECS | ID: ibc-177518

ABSTRACT

INTRODUCCIÓN Y OBJETIVOS: La evidencia de la eficacia y seguridad de la anticoagulación oral con dicumarínicos en pacientes en hemodiálisis con fibrilación auricular (FA) es controvertida. El objetivo de nuestro estudio es evaluar las implicaciones a nivel pronóstico a largo plazo de la anticoagulación con dicumarínicos en una cohorte de pacientes con FA no valvular en programa de hemodiálisis debido a insuficiencia renal terminal. MÉTODOS: Estudio observacional retrospectivo con inclusión consecutiva de 74 pacientes en hemodiálisis con FA. El periodo de inclusión fue de enero de 2005 a octubre de 2016. Las variables principales fueron mortalidad por todas las causas, reingresos no programados y sangrados. RESULTADOS: La edad media fue de 75 ± 10 años; el 66,2% fueron hombres y 43 pacientes (58,1%) recibieron acenocumarol. Durante una mediana de seguimiento de 2,40 años (IQR = 0,88-4,15), el acenocumarol no demostró beneficio en supervivencia [HR = 0,76, IC 95% (0,35-1,66), p = 0,494]. Sin embargo, los pacientes anticoagulados presentaron más riesgo de hospitalizaciones cardiovasculares recurrentes [IRR=3,94, IC 95% (1,06-14,69), p = 0,041]. Hubo una tendencia a un aumento de hospitalizaciones repetidas de causa isquémica en los pacientes anticoagulados [IRR = 5,80, IC 95% (0,86-39,0), p = 0,071]. Se observó una tendencia estadística hacia un mayor riesgo de sangrados totales recurrentes en los anticoagulados [IRR = 4,43, IC 95% (0,94-20,81), p = 0,059]. CONCLUSIONES: En el presente estudio, la anticoagulación oral con acenocumarol en pacientes en hemodiálisis con FA no supuso un aumento de la supervivencia, y sin embargo, se asoció con un mayor riesgo de hospitalizaciones de causa cardiovascular y una tendencia a mayor riesgo de sangrados totales


INTRODUCTION AND OBJECTIVES: Evidence for the efficacy and safety of oral anticoagulation with dicumarines in patients with atrial fibrillation (AF) on hemodialysis is controversial. The aim of our study is to evaluate the long-term prognostic implications of anticoagulation with dicumarines in a cohort of patients with non-valvular AF on a hemodialysis program due to end-stage renal disease. METHODS: Retrospective, observational study with consecutive inclusion of 74 patients with AF on hemodialysis. The inclusion period was from January 2005 to October 2016. The primary variables were all-cause mortality, non-scheduled readmissions and bleeding during follow-up. RESULTS: Mean age was 75 ± 10 years; 66.2% were men and 43 patients (58.1%) received acenocoumarol. During a median follow-up of 2.40 years (IQR = 0.88-4.15), acenocoumarol showed no survival benefit [HR = 0.76, 95% CI (0.35-1.66), p = 0.494]. However, anticoagulated patients were at increased risk of recurrent cardiovascular hospitalizations [IRR = 3.94, 95% CI (1.06-14.69), p = 0.041]. There was a trend towards an increase in repeated hospitalizations of ischemic cause in anticoagulated patients [IRR = 5.80, 95% CI (0.86-39.0), p = 0.071]. There was a statistical trend towards a higher risk of recurrent total bleeding in patients treated with acenocoumarol [IRR = 4.43, 95% CI (0.94-20.81), p = 0.059]. CONCLUSIONS: In this study, oral anticoagulation with acenocoumarol in patients with AF on hemodialysis did not increase survival. However, it was associated with an increased risk of hospitalizations of cardiovascular causes and a tendency to an increased risk of total bleeding


Subject(s)
Humans , Male , Female , Aged , Atrial Fibrillation/drug therapy , Anticoagulants/therapeutic use , Kidney Failure, Chronic/therapy , Renal Dialysis , Atrial Fibrillation/mortality , Follow-Up Studies , Time Factors , Prognosis
14.
Nefrologia (Engl Ed) ; 38(4): 394-400, 2018.
Article in English, Spanish | MEDLINE | ID: mdl-29426785

ABSTRACT

INTRODUCTION AND OBJECTIVES: Evidence for the efficacy and safety of oral anticoagulation with dicumarines in patients with atrial fibrillation (AF) on hemodialysis is controversial. The aim of our study is to evaluate the long-term prognostic implications of anticoagulation with dicumarines in a cohort of patients with non-valvular AF on a hemodialysis program due to end-stage renal disease. METHODS: Retrospective, observational study with consecutive inclusion of 74 patients with AF on hemodialysis. The inclusion period was from January 2005 to October 2016. The primary variables were all-cause mortality, non-scheduled readmissions and bleeding during follow-up. RESULTS: Mean age was 75±10 years; 66.2% were men and 43 patients (58.1%) received acenocoumarol. During a median follow-up of 2.40 years (IQR=0.88-4.15), acenocoumarol showed no survival benefit [HR=0.76, 95% CI (0.35-1.66), p=0.494]. However, anticoagulated patients were at increased risk of recurrent cardiovascular hospitalizations [IRR=3.94, 95% CI (1.06-14.69), p=0.041]. There was a trend towards an increase in repeated hospitalizations of ischemic cause in anticoagulated patients [IRR=5.80, 95% CI (0.86-39.0), p=0.071]. There was a statistical trend towards a higher risk of recurrent total bleeding in patients treated with acenocoumarol [IRR=4.43, 95% CI (0.94-20.81), p=0.059]. CONCLUSIONS: In this study, oral anticoagulation with acenocoumarol in patients with AF on hemodialysis did not increase survival. However, it was associated with an increased risk of hospitalizations of cardiovascular causes and a tendency to an increased risk of total bleeding.


Subject(s)
Acenocoumarol/therapeutic use , Anticoagulants/therapeutic use , Atrial Fibrillation/complications , Kidney Failure, Chronic/complications , Kidney Failure, Chronic/therapy , Renal Dialysis , Aged , Female , Humans , Male , Prognosis , Retrospective Studies , Time Factors
15.
J Proteomics ; 165: 102-112, 2017 08 08.
Article in English | MEDLINE | ID: mdl-28625740

ABSTRACT

Gluten proteins are major determinants of the bread making quality of wheat, but also of important wheat-related disorders, including coeliac disease (CD), and allergies. We carried out a proteomic study using the total grain proteins from two low-gliadin wheat lines, obtained by RNAi, and the untransformed wild type as reference. The impact of silencing on both target and on non-target proteins was evaluated. Because of the great protein complexity, we performed separate analyses of four kernel protein fractions: gliadins and glutenin subunits, and metabolic and CM-like proteins, by using a classical 2D electrophoresis gel based approach followed by RP-HPLC/nESI-MS/MS. As a result of the strong down-regulation of gliadins, the HMW-GS, metabolic and chloroform/methanol soluble proteins were over-accumulated in the transgenic lines, especially in the line D793, which showed the highest silencing of gliadins. Basing on these data, and considering that metabolic proteins and chloroform/methanol soluble proteins (CM-like), such as the α-amylase/trypsin inhibitor family, ß-amylase and serpins, were related to wheat allergens, further in vivo analysis will be needed, especially those related to clinical trials in controlled patients, to determine if these lines could be used for food preparation for celiac or other gluten intolerant groups. BIOLOGICAL SIGNIFICANCE: Several enteropathies and allergies are related to wheat proteins. Biotechnological techniques such as genetic transformation and RNA interference have allowed the silencing of gliadin genes, providing lines with very low gliadin content in the grains. We report a proteomic-based approach to characterize two low-gliadin transgenic wheat lines obtained by RNAi technology. These lines harbor the same silencing fragment, but driven by two different endosperm specific promoters (γ-gliadin and D-hordein). The comprehensive proteome analysis of these transgenic lines, by combining two-dimensional electrophoresis and RP-HPLC/nESI-MS/MS, provided a large number of spots differentially expressed between the control and the transgenic lines. Hence, the results of this study will facilitate further safety evaluation of these transgenic lines.


Subject(s)
Gliadin/genetics , Plants, Genetically Modified , Proteomics/methods , Triticum/chemistry , Bread , Chromatography, High Pressure Liquid , Electrophoresis, Gel, Two-Dimensional , Gene Silencing , Plant Proteins/analysis , Tandem Mass Spectrometry , Triticum/genetics
16.
Development ; 144(12): 2270-2281, 2017 06 15.
Article in English | MEDLINE | ID: mdl-28507001

ABSTRACT

Dorsal-ventral axis formation in the sea urchin embryo relies on the asymmetrical expression of the TGFß Nodal. The p38-MAPK pathway has been proposed to be essential for dorsal-ventral axis formation by acting upstream of nodal expression. Here, we report that, in contrast to previous studies that used pharmacological inhibitors of p38, manipulating the activity of p38 by genetic means has no obvious impact on morphogenesis. Instead, we discovered that p38 inhibitors strongly disrupt specification of all germ layers by blocking signalling from the Nodal receptor and by interfering with the ERK pathway. Strikingly, while expression of a mutant p38 that is resistant to SB203580 did not rescue dorsal-ventral axis formation or skeletogenesis in embryos treated with this inhibitor, expression of mutant Nodal receptors that are resistant to SB203580 fully restored nodal expression in SB203580-treated embryos. Taken together, these results establish that p38 activity is not required for dorsal-ventral axis formation through nodal expression nor for skeletogenesis. Our results prompt a re-evaluation of the conclusions of several recent studies that linked p38 activity to dorsal-ventral axis formation and to patterning of the skeleton.


Subject(s)
Paracentrotus/embryology , Paracentrotus/enzymology , p38 Mitogen-Activated Protein Kinases/metabolism , Amino Acid Sequence , Animals , Body Patterning/drug effects , Body Patterning/genetics , Body Patterning/physiology , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Imidazoles/pharmacology , MAP Kinase Signaling System/drug effects , Morphogenesis/drug effects , Morphogenesis/genetics , Morphogenesis/physiology , Mutation , Nodal Signaling Ligands/genetics , Nodal Signaling Ligands/metabolism , Paracentrotus/genetics , Phenotype , Protein Kinase Inhibitors/pharmacology , Pyridines/pharmacology , Sequence Homology, Amino Acid , Signal Transduction/drug effects , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , p38 Mitogen-Activated Protein Kinases/genetics
17.
Front Plant Sci ; 8: 257, 2017.
Article in English | MEDLINE | ID: mdl-28289425

ABSTRACT

Gluten proteins are major determinants of the bread making quality of wheat but also of important gluten-related disorders. The gluten protein accumulation during grain filling is strongly influenced by nitrogen fertilization. We have characterized the gluten proteins in low-gliadin wheat lines as influenced by nitrogen treatments in two experiments. These transgenic lines, D783, D793, C655, D577, and E82 were obtained by using two different RNAi silencing fragments and two endosperm-specific promoters to drive the silencing fragments (d-hordein and γ-gliadin). In Experiment 1, we used three nitrogen fertilizer rates (120, 360, and 1080 mg N) added at sowing stage and combined with two sulfur rates (8 and 30 mg S); Experiment 2 included two nitrogen levels (120 and 1080 mg N), which were added according to the greatest demand per plant using split applications. The protein quantification was accomplished by Reverse-Phase High-Performance Liquid Chromatography and gluten content (ppm) determined using monoclonal antibody R5 (Competitive R5 ELISA). The results showed differences in protein accumulation between the two transgenic lines with the same silencing fragment but different promoter. Lines D793 and E82 showed low gliadin and an increment in glutenin content with increasing nitrogen. Competitive ELISA R5 showed a significant decrease in gluten content using split applications of nitrogen (Experiment 2) with 120 mg N compared to Experiment 1. In addition, line E82 ensures that variations in N fertilization will not result in increased gluten content.

18.
Aten. prim. (Barc., Ed. impr.) ; 48(10): 632-641, dic. 2016. tab, ilus
Article in English | IBECS | ID: ibc-158663

ABSTRACT

AIM: To determine the communicative profiles of family physicians and the characteristics associated with an improved level of communication with the patient. DESIGN: A descriptive multicentre study. LOCATION: Primary Healthcare Centres in Almeria, Granada, Jaen and Huelva. PARTICIPANT:119 family physicians (tutors and 4th year resident physicians) filmed and observed with patients. Principal measurements: Demographic and professional characteristics. Analysis of the communication between physicians and patients, using a CICAA (Connect, Identify, Understand, Agree and Assist, in English) scale. A descriptive, bivariate, multiple linear regression analysis was performed. RESULTS: There were 436 valid interviews. Almost 100% of physicians were polite and friendly, facilitating a dialogue with the patient and allowing them to express their doubts. However, few physicians attempted to explore the state of mind of the patient, or enquire about their family situation or any important stressful events, nor did they ask open questions. Furthermore, few physicians summarised the information gathered. The mean score was 21.43±5.91 points (maximum 58). There were no differences in the total score between gender, city, or type of centre. The linear regression verified that the highest scores were obtained from tutors (B: 2.98), from the duration of the consultations (B: 0.63), and from the age of the professionals (B: −0.1). CONCLUSION: Physicians excel in terms of creating a friendly environment, possessing good listening skills, and providing the patient with information. However the ability to empathise, exploring the psychosocial sphere, carrying out shared decision-making, and asking open questions must be improved. Being a tutor, devoting more time to consultations, and being younger, results in a significant improvement in communication with the patienT


OBJETIVO: Conocer el perfil comunicacional de los médicos de familia y las caracteristicas asociadas a una mejor comunicación con el paciente. DISEÑO: Estudio descriptivo multicéntrico. Emplazamiento. Centros de salud de atención primaria de Almería, Granada, Jaén y Huelva. PARTICIPANTES: Ciento diecinueve médicos de familia (tutores y residentes de 4. ° año) videograbados en consulta. MEDICIONES PRINCIPALES: Características demográficas y profesionales. Análisis de la comunicación médico-paciente mediante la escala Conectar, Identificar y Comprender, Acordar y Ayudar (CICAA). Se realizó un análisis descriptivo, bivariable y de regresión lineal múltiple. RESULTADOS: Se obtuvieron 436 entrevistas válidas. Casi el 100% se muestran corteses y amables, facilitan el discurso del paciente y permiten que exprese sus dudas. En cambio, pocos profesionales resumen la información, exploran el estado de ánimo del paciente, su entorno familiar, acontecimientos vitales estresantes o emplean preguntas abiertas. La puntuación media fue de 21.43±5.91 puntos (máximo 58). No existieron diferencias en la puntuación por sexo, ciudad o tipo de centro. Mediante regresión lineal múltiple se comprueba que una mayor puntuación se relaciona con ser tutor (B:2.98), el tiempo de consulta (B:0.63) y la edad del profesional (B: −0,1). CONCLUSIONES: Los médicos destacan por crear un clima cálido, buena escucha e informar al paciente; en cambio, deberían mejorarse la empatía, la exploración de la esfera psicosocial, realizar preguntas abiertas y la toma de decisiones compartidas. Ser tutor, mayor tiempo de consulta y ser más joven se relacionan con una mejor comunicación con el paciente (AU)


Subject(s)
Humans , Male , Female , Aptitude , Health Knowledge, Attitudes, Practice , Internship and Residency , Physicians, Family/organization & administration , Physicians, Family/trends , Primary Health Care , Referral and Consultation/trends , Physician-Patient Relations , Communication , Family Practice , Linear Models , Health Surveys/methods , Data Analysis/methods , Analysis of Variance
19.
Aten Primaria ; 48(10): 632-641, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27157118

ABSTRACT

AIM: To determine the communicative profiles of family physicians and the characteristics associated with an improved level of communication with the patient. DESIGN: A descriptive multicentre study. LOCATION: Primary Healthcare Centres in Almeria, Granada, Jaen and Huelva. PARTICIPANTS: 119 family physicians (tutors and 4th year resident physicians) filmed and observed with patients. PRINCIPAL MEASUREMENTS: Demographic and professional characteristics. Analysis of the communication between physicians and patients, using a CICAA (Connect, Identify, Understand, Agree and Assist, in English) scale. A descriptive, bivariate, multiple linear regression analysis was performed. RESULTS: There were 436 valid interviews. Almost 100% of physicians were polite and friendly, facilitating a dialogue with the patient and allowing them to express their doubts. However, few physicians attempted to explore the state of mind of the patient, or enquire about their family situation or any important stressful events, nor did they ask open questions. Furthermore, few physicians summarised the information gathered. The mean score was 21.43±5.91 points (maximum 58). There were no differences in the total score between gender, city, or type of centre. The linear regression verified that the highest scores were obtained from tutors (B: 2.98), from the duration of the consultations (B: 0.63), and from the age of the professionals (B: -0.1). CONCLUSION: Physicians excel in terms of creating a friendly environment, possessing good listening skills, and providing the patient with information. However the ability to empathise, exploring the psychosocial sphere, carrying out shared decision-making, and asking open questions must be improved. Being a tutor, devoting more time to consultations, and being younger, results in a significant improvement in communication with the patient.


Subject(s)
Communication , Family Practice/education , Internship and Residency , Physician-Patient Relations , Primary Health Care , Adult , Female , Health Facilities , Humans , Male , Middle Aged
20.
PLoS One ; 11(3): e0152292, 2016.
Article in English | MEDLINE | ID: mdl-27018786

ABSTRACT

SCOPE: The aim of this work was to assess the ability of Near Infrared Spectroscopy (NIRS) to distinguish wheat lines with low gliadin content, obtained by RNA interference (RNAi), from non-transgenic wheat lines. The discriminant analysis was performed using both whole grain and flour. The transgenic sample set included 409 samples for whole grain sorting and 414 samples for flour experiments, while the non-transgenic set consisted of 126 and 156 samples for whole grain and flour, respectively. METHODS AND RESULTS: Samples were scanned using a Foss-NIR Systems 6500 System II instrument. Discrimination models were developed using the entire spectral range (400-2500 nm) and ranges of 400-780 nm, 800-1098 nm and 1100-2500 nm, followed by analysis of means of partial least square (PLS). Two external validations were made, using samples from the years 2013 and 2014 and a minimum of 99% of the flour samples and 96% of the whole grain samples were classified correctly. CONCLUSIONS: The results demonstrate the ability of NIRS to successfully discriminate between wheat samples with low-gliadin content and wild types. These findings are important for the development and analysis of foodstuff for celiac disease (CD) patients to achieve better dietary composition and a reduction in disease incidence.


Subject(s)
Celiac Disease , Food Analysis/methods , Gliadin/analysis , Spectroscopy, Near-Infrared , Triticum/chemistry , Diet, Gluten-Free , Discriminant Analysis , Enzyme-Linked Immunosorbent Assay , Food Analysis/standards , Gliadin/metabolism , Humans , Least-Squares Analysis , Plants, Genetically Modified/metabolism , RNA Interference
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