Integrative genetic analysis illuminates ALS heritability and identifies risk genes.

Amyotrophic lateral sclerosis (ALS) has substantial heritability, in part shared with fronto-temporal dementia (FTD). We show that ALS heritability is enriched in splicing variants and in binding sites of 6 RNA-binding proteins including TDP-43 and FUS. A transcriptome wide association study (TWAS) identified 6 loci associated with ALS, including in NUP50 encoding for the nucleopore basket protein NUP50. Independently, rare variants in NUP50 were associated with ALS risk (P = 3.71.10-03; odds ratio = 3.29; 95%CI, 1.37 to 7.87) in a cohort of 9,390 ALS/FTD patients and 4,594 controls. Cells from one patient carrying a NUP50 frameshift mutation displayed a decreased level of NUP50. Loss of NUP50 leads to death of cultured neurons, and motor defects in Drosophila and zebrafish. Thus, our study identifies alterations in splicing in neurons as critical in ALS and provides genetic evidence linking nuclear pore defects to ALS.

Role of Oxidative Stress and Lipid Peroxidation in the Pathophysiology of NAFLD.

Non-alcoholic fatty liver disease (NAFLD) is characterised by an excess of hepatic fat that can progress to steatohepatitis, fibrosis, cirrhosis and hepatocarcinoma. The imbalance between lipid uptake/lipogenesis and lipid oxidation/secretion in the liver is a major feature of NAFLD. Given the lack of a non-invasive and reliable methods for the diagnosis of non-alcoholic steatohepatitis (NASH), it is important to find serum markers that are capable of discriminating or defining patients with this stage of NASH. Blood samples were obtained from 152 Caucasian subjects with biopsy-proven NAFLD due to persistently elevated liver enzyme levels. Metabolites representative of oxidative stress were assessed. The findings derived from this work revealed that NAFLD patients with a NASH score of ≥ 4 showed significantly higher levels of lipid peroxidation (LPO). Indeed, LPO levels above the optimal operating point (OOP) of 315.39 µM are an independent risk factor for presenting a NASH score of ≥ 4 (OR: 4.71; 95% CI: 1.68−13.19; p = 0.003). The area under the curve (AUC = 0.81, 95% CI = 0.73−0.89, p < 0.001) shows a good discrimination ability of the model. Therefore, understanding the molecular mechanisms underlying the basal inflammation present in these patients is postulated as a possible source of biomarkers and therapeutic targets in NASH.

Anatomical and genetic bases underlying the convergent evolution of fleshy and dry dehiscent fruits in Cestrum and Brugmansia (Solanaceae).

The mechanisms controlling evolutionary shifts between dry and fleshy fruits in angiosperms are poorly understood. In Solanaceae, Cestrum and Brugmansia represent cases of convergent evolution of fleshy and dry fruits, respectively. Here we study the anatomical and genetic bases of the independent origin of fleshy fruits in Cestrum and the reversion to dry dehiscent fruits in Brugmansia. We also characterize the expression of candidate fruit development genes, including ALCATRAZ/SPATULA, FRUITFULL, HECATE1/2/3, REPLUMLESS and SHATTERPROOF. We identify anatomical changes to establish developmental stages in the ovary-to-fruit transition in Cestrum nocturnum and Brugmansia suaveolens. We generate reference transcriptomes for both species, isolate homologs for all genes in the fruit genetic regulatory network (GRN) and perform gene expression analyses for ALC/SPT, FUL, HEC1/2/3, RPL and SHP throughout fruit development. Finally, we compare our results to expression patterns found in typical capsules of Nicotiana tabacum and berries of Solanum lycopersicum available in public repositories. We have identified homologous, homoplasious and unique anatomical features in C. nocturnum and B. suaveolens fruits, resulting in their final appearance. Expression patterns suggest that FUL, SHP and SPT might control homologous characteristics, while ALC and RPL likely contribute to homoplasious anatomical features. The fruit GRN changes considerably in these genera when compared to typical capsules and berries of Solanaceae, particularly in B. suaveolens, where expression of FUL2 and RPL1 is lacking.

Evolution of Class II TCP genes in perianth bearing Piperales and their contribution to the bilateral calyx in Aristolochia.

Controlled spatiotemporal cell division and expansion are responsible for floral bilateral symmetry. Genetic studies have pointed to class II TCP genes as major regulators of cell division and floral patterning in model core eudicots. Here we study their evolution in perianth-bearing Piperales and their expression in Aristolochia, a rare occurrence of bilateral perianth outside eudicots and monocots. The evolution of class II TCP genes reveals single-copy CYCLOIDEA-like genes and three paralogs of CINCINNATA (CIN) in early diverging angiosperms. All class II TCP genes have independently duplicated in Aristolochia subgenus Siphisia. Also CIN2 genes duplicated before the diversification of Saruma and Asarum. Sequence analysis shows that CIN1 and CIN3 share motifs with Cyclin proteins and CIN2 genes have lost the miRNA319a binding site. Expression analyses of all paralogs of class II TCP genes in Aristolochia fimbriata point to a role of CYC and CIN genes in maintaining differential perianth expansion during mid- and late flower developmental stages by promoting cell division in the distal and ventral portion of the limb. It is likely that class II TCP genes also contribute to cell division in the leaf, the gynoecium and the ovules in A. fimbriata.

Expression and function of the bHLH genes ALCATRAZ and SPATULA in selected Solanaceae species.

The genetic mechanisms underlying fruit development have been identified in Arabidopsis and have been comparatively studied in tomato as a representative of fleshy fruits. However, comparative expression and functional analyses on the bHLH genes downstream the genetic network, ALCATRAZ (ALC) and SPATULA (SPT), which are involved in the formation of the dehiscence zone in Arabidopsis, have not been functionally studied in the Solanaceae. Here, we perform detailed expression and functional studies of ALC/SPT homologs in Nicotiana obtusifolia with capsules, and in Capsicum annuum and Solanum lycopersicum with berries. In Solanaceae, ALC and SPT genes are expressed in leaves, and all floral organs, especially in petal margins, stamens and carpels; however, their expression changes during fruit maturation according to the fruit type. Functional analyses show that downregulation of ALC/SPT genes does not have an effect on gynoecium patterning; however, they have acquired opposite roles in petal expansion and have been co-opted in leaf pigmentation in Solanaceae. In addition, ALC/SPT genes repress lignification in time and space during fruit development in Solanaceae. Altogether, some roles of ALC and SPT genes are different between Brassicaceae and Solanaceae; while the paralogs have undergone some subfunctionalization in the former they are mostly redundant in the latter.

Role of the PNPLA3 polymorphism rs738409 on silymarin + vitamin E response in subjects with non-alcoholic fatty liver disease.

BACKGROUND: non-alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease in developed countries. Lifestyle changes are the pillar of the treatment, although a pharmacological approach is sometimes required in the case of a failure to respond/adhere to the diet. OBJECTIVE: the aim of this study was to evaluate the effect of silymarin and the influence of the patatin-like phospholipase domain-containing protein 3 (PNPLA3) variant on the response to treatment in patients with NAFLD in a pilot study. METHODS: a total of 54 patients with a NAFLD proven biopsy were enrolled in an open prospective study and were treated with Eurosil 85® (silymarin + vitamin E) for six months. Biochemical parameters and cardiovascular risk factors (diabetes mellitus, dyslipidemia, hypertriglyceridemia, arterial hypertension and HOMA-IR > 2.5) were recorded before and after six months of treatment. Non-invasive indexes (fatty liver index, lipid accumulation product and NAFLD-fibrosis score) were also calculated. The rs738409 PNPLA3 gene polymorphism status was also determined. RESULTS: significant statistical changes from baseline values after six months of treatment were observed in transaminases levels but not in non-invasive index markers. Twenty patients (37.1%) were G allele carriers and had a higher percentage of lobular inflammation and ballooning on the basal liver biopsy. Patients with the G allele had a smaller decrease in transaminases levels after treatment with silymarin + vitamin E than non-G-allele carriers. CONCLUSIONS: treatment with silymarin + vitamin E produced a decrease in transaminases after six months of treatment without an accompanying weight loss. PNPLA3 G-allele carriers responded poorly to the treatment.

Analytical, anthropometric and dietary factors associated with the development of fibrosis in patients with nonalcoholic fatty liver disease.

BACKGROUND: a prolonged non-alcoholic steatohepatitis (NASH) condition can lead to advanced stages of liver disease and the development of hepatocellular carcinoma. AIM: to evaluate analytical, anthropometric and dietary factors associated with the presence of fibrosis as this is the factor that most influences survival and evolution. METHODS: seventy-six patients with liver biopsy-diagnosed non-alcoholic fatty liver disease (NAFLD) were included. Biopsies were scored considering the NASH criteria of Kleiner. Analytical, anthropometric and dietary (survey) parameters were obtained. NAFLD-FS is a non-invasive fibrosis index and was assessed for each patient. Leptin, adiponectin, resistin and TNF-alpha serum levels were determined. RESULTS: fifty-six patients were male (73.7%) and the mean age was 44.5 ± 11.3 years of age (19-68). Thirty-nine (51.3%) (F1-F2: 84.6%; F3-4: 15.4%) patients had fibrosis in the liver biopsy. Seventeen females (85%) had fibrosis versus 22 males (39%), which was statistically significant by univariate analysis (p < 0.01). Patients with advanced fibrosis were older, with lower platelet counts, lower serum albumin, greater homeostatic model assessment insulin resistance (HOMA-IR), lower dietary lipids percentage, higher serum leptin levels and higher NAFLD Fibrosis Score (NAFLD-FS) values. This index had a negative predictive value of 98% and a positive predictive value of 60% for the detection of fibrosis. Variables independently associated with fibrosis (logistic regression) included male gender (protective factor) (0.09, 95% CI 0.01-0.7; p < 0.05) and HOMA-IR (1.7, 95% CI, 1.03-2.79; p < 0.05). CONCLUSIONS: gender and HOMA-IR were the only independent factors associated with fibrosis. NAFLD-FS could be considered as an accurate scoring system to rule out advanced fibrosis.

Evolution and Expression Patterns of TCP Genes in Asparagales.

CYCLOIDEA-like genes are involved in the symmetry gene network, limiting cell proliferation in the dorsal regions of bilateral flowers in core eudicots. CYC-like and closely related TCP genes (acronym for TEOSINTE BRANCHED1, CYCLOIDEA, and PROLIFERATION CELL FACTOR) have been poorly studied in Asparagales, the largest order of monocots that includes both bilateral flowers in Orchidaceae (ca. 25.000 spp) and radially symmetrical flowers in Hypoxidaceae (ca. 200 spp). With the aim of assessing TCP gene evolution in the Asparagales, we isolated TCP-like genes from publicly available databases and our own transcriptomes of Cattleya trianae (Orchidaceae) and Hypoxis decumbens (Hypoxidaceae). Our matrix contains 452 sequences representing the three major clades of TCP genes. Besides the previously identified CYC specific core eudicot duplications, our ML phylogenetic analyses recovered an early CIN-like duplication predating all angiosperms, two CIN-like Asparagales-specific duplications and a duplication prior to the diversification of Orchidoideae and Epidendroideae. In addition, we provide evidence of at least three duplications of PCF-like genes in Asparagales. While CIN-like and PCF-like genes have multiplied in Asparagales, likely enhancing the genetic network for cell proliferation, CYC-like genes remain as single, shorter copies with low expression. Homogeneous expression of CYC-like genes in the labellum as well as the lateral petals suggests little contribution to the bilateral perianth in C. trianae. CIN-like and PCF-like gene expression suggests conserved roles in cell proliferation in leaves, sepals and petals, carpels, ovules and fruits in Asparagales by comparison with previously reported functions in core eudicots and monocots. This is the first large scale analysis of TCP-like genes in Asparagales that will serve as a platform for in-depth functional studies in emerging model monocots.

ß amyloid protein precursor-like (Appl) is a Ras1/MAPK-regulated gene required for axonal targeting in Drosophila photoreceptor neurons.

In a genome-wide expression profile search for genes required for Drosophila R7 photoreceptor development we found ß amyloid protein precursor-like (Appl), the ortholog of human APP, which is a key factor in the pathogenesis of Alzheimer's disease. We analyzed Appl expression in the eye imaginal disc and found that is highly accumulated in R7 photoreceptor cells. The R7 photoreceptor is responsible for UV light detection. To explore the link between high expression of Appl and R7 function, we have analyzed Appl null mutants and found reduced preference for UV light, probably because of mistargeted R7 axons. Moreover, axon mistargeting and inappropriate light discrimination are enhanced in combination with neurotactin mutants. R7 differentiation is triggered by the inductive interaction between R8 and R7 precursors, which results in a burst of Ras1/MAPK, activated by the tyrosine kinase receptor Sevenless. Therefore, we examined whether Ras1/MAPK is responsible for the high Appl expression. Inhibition of Ras1 signaling leads to reduced Appl expression, whereas constitutive activation drives ectopic Appl expression. We show that Appl is directly regulated by the Ras/MAPK pathway through a mechanism mediated by PntP2, an ETS transcription factor that specifically binds ETS sites in the Appl regulatory region. We also found that zebrafish appb expression increased after ectopic fgfr activation in the neural tube of zebrafish embryos, suggesting a conserved regulatory mechanism.

Comparative anatomical and developmental analysis of dry and fleshy fruits of Solanaceae.

PREMISE OF THE STUDY: An anatomical examination of dry and fleshy fruits within the Solanaceae was carried out to identify comparable stages throughout development as well as features exclusive to each type of fruit. We studied fruit development of Nicotiana and Petunia, which have the plesiomorphic capsular fruit; Solanum and Iochroma, characterized by a derived fleshy berry; Cestrum, an independent origin of a fleshy fruit; and Datura, a reversion to a dry fruit. • METHODS: Pre- and postanthesis carpels and fruits of all species were collected, sectioned, stained, and examined using light microscopy. • KEY RESULTS: Comparable stages of carpel and fruit development were identified in all species. Furthermore, anatomical and developmental features were identified that characterize capsules in Solanaceae, including lack of increase in the number of pericarp cell layers, formation of a sclerified endocarp, and elongation of the epidermal cells of the placenta. Pericarps of fleshy fruits of the Solanoideae are characterized by abundant collenchyma, an increase in the number of cell layers, and a parenchymatous endocarp often expanding into the locules. Anatomical data show that early developmental stages of the fruit of Cestrum, a berry, are similar to the capsular fruits of Petunia and Nicotiana; similarly, Datura, one of the few capsular members of the Solanoideae, shares several anatomical features with closely related berried taxa. • CONCLUSIONS: Ontogenetically, all fleshy or all dry fruits do not necessarily share a common developmental ground plan. Independent evolution of fleshiness, sclerification, dryness, and dehiscence are discussed in a phylogenetic context.

Parte iii de hesslow a hooker: darwin y los inicios del pensamiento evolutivo en botánica / From Henslow to Hooker: Darwin and the Early Evolutionary Thought in Botany

Aunque no lo suficientemente conocida y difundida, la obra botánica de Darwin aportó una gran cantidad de evidencia empírica fundamental para el establecimiento de la revolución darwinista. Se describe el desarrollo de esta obra, en especial con relación a los dos mentores de Darwin en botánica: J. S. Henslow y J. D. Hooker. Además de numerosos artículos y notas en sus diarios de viaje, su correspondencia y numerosos apartes de sus dos obras magnas El origen de las especies y selección natural, Darwin escribió siete libros relacionados con diversos aspectos de la botánica, incluída la polinización en orquídeas, la morfología y fisiología de plantas trepadoras, la domesticación, las plantas insectívoras, la polinización, las formas florales, y los movimientos de las plantas. Cada uno de estos libros es ahora clásico en cada tema. La introducción de la teoría evolutiva en la sistemática de plantas enriqueció los distintos sistemas de clasificación en los 70 años que siguieron a la publiación de El origen, lo cual está en estrecha relación con las preguntas, aún vigentes, acerca del origen y la diversificación temprana de las angiospermas. A la vez, se revisa la influencia de las contribuciones botánicas de Darwin en las obras de autores en diversos países de Europa y América, y en disciplinas tan diversas y actuales como la biogeografía, la biología reproductiva en muy diversos grupos de plantas con flor, la citología y mecanismos de herencia en la célula vegetal, la teratología vegetal, las variaciones debidas a domesticación, y la reciente integración de evolución, genética y desarrollo en la disciplina conocida como evo-devo.

Despite Darwin s botanical works are not sufficiently known, they provided a large amount of critical, empirical evidence in favor of the darwinian revolution. This paper describes the development of such works in connection to the influence of two of Darwin s mentors in botany, J. S. Henslow and J. D. Hooker. Besides numerous articles, field notes, extensive correspondence, and Darwin s main books, The Origin of Species and Natural Selection, he wrote seven books related to different botanical issues, including orchid pollination, morphology and physiology of climbers, domestication, insectivorous plants, cross-and self-pollination, floral forms, and plant movements. Each of these volumes became classic on each topic. The introduction of the evolutionary theory in plant systematics had a positive effect by increasing the systems of classification during the 70 years following the publication of The Origin, which is in close relation to the still unresolved questions about the origin and early diversification of angiosperms. In addition, we review the influence of Darwin s contributions in European and American authors, as well as in current disciplines such as biogeography, reproductive biology, citology and mechanisms of heredity in the plant cell, plant teratology, plant domestication, and the new integration of evolution, genetics and development under the discipline known as evo-devo.

El desarollo sociomoral de adolescentes de Costa Rica según el modelo de GIBBS / Socio-moral development among Costa Rican adolescents according to the Gibb's model

This study examines the reliability of the Questionnaire of Sociomoral Reflection in Costa Rican version, elaborated originally by John C. Gibbs and adapted by the author. The questionnaire is applied in short form. By means of three studies, with different samples, interindividual differences are evaluated, in age and gender variables, in sociomoral development of Costa Rican adolescents. The results show appropriate reliability coefficients in three studies. Also, it is carried out a preliminary validity analysis of questionnaire. Results allow to verify clear tendencies informed in the literature about the topic in gender individual differences. For variable age, there are results evidencing a clear developmental trend according to which the sociomoral development seems to show a narrow dependence of the influence of educational level of those who participate in this research.

Este estudio examina la fiabilidad del "Cuestionario de Reflexión Sociomoral" en su versión costarricense, elaborado originalmente por John C. Gibbs. El cuestionario es aplicado en su forma corta y mediante tres estudios, con muestras distintas, son evaluadas las diferencias interindividuales, en las variables de edad y género, en el desarrollo sociomoral de adolescentes costarricenses. Los resultados muestran adecuados índices de fiabilidad en los tres estudios. Además, se lleva a cabo un análisis preliminar de validez del cuestionario. En cuanto a las diferencias individuales por género, los resultados permiten verificar tendencias claras informadas en la literatura acerca del tema. En cuanto a la variable edad, hay resultados evidenciando una clara tendencia evolutiva según la cual el desarrollo sociomoral parece mostrar una estrecha dependencia de la influencia del nivel educativo de quienes participan en esta investigación.

Implicaciones médico-legales de las perforaciones corporales / Medical-legal implications of body piercings

La perforación corporal es una actividad cada vez más común en nuestro medio, por lo tanto es de importancia tanto para el médico como para el especialista en medicina legal pues ya se les exige que tengan un amplio conocimiento acerca de la técnica correcta de la perforación corporal, de los cuidados necesarios y las principales complicaciones de los piercings. En la prática hospitalaria hoy día es frecuente encontrarse con personas que consultan ya sea porque desean colocarse un piercing y quieren escuchar un consejo de su médico o personas que consultan a causa de las múltiples complicaciones que pueden ocurrir.