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Several impacts over ecosystem services have been produced by land use/cover changes, placing it as one of the main factors driving global environmental change. In the present study, the SWAT model was used to assess the effect of land use/cover changes on the hydrology response in the Andalien river basin from the south-central zone of Chile. Three land use/cover scenarios (LU_1986, LU_2001, and LU_2011) were compared over a period of 30 years (1984−2013) to remove the effect of climate variability on hydrology. The results show a significant decrease in total annual flows among the three LU scenarios. The greater differences in the annual flows of 25.05 m3/s were observed between LU_1986 and LU_2011 scenarios. The hydrological cycle dynamics in the basin show an increasing trend of evapotranspiration and surface flows with a significant decrease in percolation and lateral flow on a monthly and seasonal scale. This behavior can be explained by the increasing percentage of the basin area covered by exotic plantations, from 35.22% to 63.93% during the period. The evidence of these changes and the evaluation of their effects are particularly relevant for the long-term sustainable management of water resources.

Summary: Paxillin is a scaffold protein that participates in focal adhesion signaling in mammalian cells. Fission yeast paxillin ortholog, Pxl1, is required for contractile actomyosin ring (CAR) integrity and collaborates with the β-glucan synthase Bgs1 in septum formation. We show here that Pxl1’s main function is to recruit calcineurin (CN) phosphatase to the actomyosin ring; and thus the absence of either Pxl1 or calcineurin causes similar cytokinesis defects. In turn, CN participates in the dephosphorylation of the Cdc15 F-BAR protein, which recruits and concentrates Pxl1 at the CAR. Our findings suggest the existence of a positive feedback loop between Pxl1 and CN and establish that Pxl1 is a crucial component of the CN signaling pathway during cytokinesis. : Martín-García et al. show that fission yeast paxillin binds and recruits calcineurin (CN) phosphatase to the cytokinetic actomyosin ring (CAR). There, CN collaborates with Bgs1 in septum ingression and promotes the concentration of Pxl1 at the CAR in a feedback loop likely mediated through CN dephosphorylation of the Cdc15 F-BAR protein, essential for cytokinesis. Keywords: Schizosaccharomyces pombe, cytokinesis, calcineurin, paxillin, septum, actomyosin ring, Bgs1, Cdc15

Introdução: O médico de família (MF) é com frequência o primeiro contacto do utente com os cuidados de saúde, lidando com a doença numa fase precoce e cujos sintomas são ainda inespecíficos. Com este caso pretende-se promover a reflexão sobre o papel do MF nas várias etapas do processo diagnóstico, bem como as características da medicina geral e familiar e dos cuidados de saúde primários que o facilitam. Descrição do caso: Mulher de 48 anos, pertencente a uma família reconstruída e índice socioeconómico III de Graffar. Tem antecedentes pessoais de síndroma depressivo (medicada com fluoxetina), encontrando-se em climatério. Recorreu à consulta por náuseas e enfartamento com dez dias de evolução, episódio de vómito isolado na véspera de Natal e prurido generalizado. Objetivamente apresentava dor discreta à palpação epigástrica. Foi medicada sintomaticamente e pediu-se avaliação analítica e ecografia abdominal. Regressou três dias depois por icterícia das escleróticas e acolia, mantendo náuseas. Negava colúria ou febre. Os exames complementares de diagnóstico vieram a revelar analiticamente uma hiperbilirrubinemia conjugada; ecograficamente apresentava marcada dilatação da via biliar principal, imagem nodular no corpo/cauda pancreáticos e múltiplos conglomerados adenopáticos adjacentes aos grandes vasos. Não apresentava sintomas consumptivos evidentes. Pediu-se tomografia computorizada abdominal que revelou conglomerado adenopático envolvendo a cabeça do pâncreas, traduzindo provável doença linfoproliferativa. Foi referenciada com caráter urgente à consulta de hemato-oncologia onde se confirmou o diagnóstico: linfoma não-Hodgkin B difuso de grandes células. Cumpriu oito ciclos de quimioterapia e atualmente encontra-se em remissão. Comentário: Apesar de a tomada de decisão do MF dever basear-se no padrão de morbilidade da comunidade e no contexto sociocultural do doente, não deve ser desvalorizada a diversidade de hipóteses de diagnóstico que poderão resultar da evolução natural da doença. O MF deve suportar-se das vantagens da continuidade e da acessibilidade aos cuidados para este processo, potenciando assim o seu papel diagnóstico.

Murine colitis models are crucial tools for understanding intestinal homeostasis and inflammation. However, most current models utilize a highly inbred strain of mice, and often only one sex is employed to limit bias. This targeted approach, which in itself is biased, means that murine genetic diversity and sex-related differences are ignored, making it even more difficult to extend findings to humans, who are highly heterogeneous. Furthermore, most models do not examine the chronic form of colitis, an important fact taking into account the chronic nature of the inflammatory bowel diseases (IBD). Here, we attempted to create a more realistic murine colitis model by addressing these three issues. Using chemically induced chronic colon inflammation in an outbred strain of mice (RjOrl:SWISS [CD-1]), we (i) mimicked the relapsing nature of the disease, (ii) better represented normal genetic variability, and (iii) employed both female and male mice. Colitis was induced by intrarectal administration of dinitrobenzene sulfonic acid (DNBS). After a recovery period and 3 days before the mice were euthanized, colitis was reactivated by a second administration of DNBS. Protocol length was 24 days. Colitis severity was assessed using body mass, macroscopic scores, and histological scores. Myeloperoxidase (MPO) activity, cytokine levels, and lymphocyte populations were also characterized. Our results show that the intrarectal administration of DNBS effectively causes colitis in both female and male CD-1 mice in a dose-dependent manner, as reflected by loss of body mass, macroscopic scores and histological scores. Furthermore, colon cytokine levels and mesenteric lymph node characteristics indicate that this model involves immune system activation. Although some variables were sex-specific, most of the results support including both females and males in the model. Our ultimate goal is to make this model available to researchers for testing candidate anti-inflammatory agents, such as classical or next-generation probiotics; we also aim for the results to be more easily transferrable to human trials.

Faecalibacterium prausnitzii represents approximately 5% of the total fecal microbiota in healthy adults being one of the most abundant bacterium in the human intestinal microbiota of healthy adults. Furthermore, this bacterium has been proposed to be a sensor and a major actor of the human intestinal health because of its importance in the gut ecosystem. In this context, F. prausnitzii population levels have been found to be reduced in patients suffering from several syndromes and diseases such as inflammatory bowel diseases. These diseases are characterized by a breakage of the intestinal homeostasis called dysbiosis and the use of F. prausnitzii as a next generation probiotic (also called live biotherapeutics) has been proposed as a natural tool to restore such dysbiosis within the gut. Nevertheless, despite the potential importance of this bacterium in human health, little is known about its main effectors underlying its beneficial effects. In this perspective note, we aim to present the actual state in the research about F. prausnitzii effectors and the future milestones in this field.

ABSTRACT Mucosa-associated lymphoid tissue (MALT) lymphoma is an extranodal lymphoma arising in a number of epithelial tissues, including stomach, salivary gland and the lung. Pulmonary MALT lymphoma originates in the parenchyma and/or bronchi and is a rare disease, representing 0.5-1% of primary pulmonary malignancies. Although the time to diagnosis can be delayed because most patients are asymptomatic and investigations are usually driven by the accidental finding of abnormal lung imaging studies, it has an indolent course and a good prognosis. Here we report the case of an asymptomatic female patient with a nodular lung opacity that was found to persist five months after an episode of community-acquired pneumonia. A final diagnosis of pulmonary MALT lymphoma was obtained after an extensive diagnostic work-up. RESUMO: O linfoma do tecido linfóide associado à mucosa (MALT) é um linfoma extranodal que surge em vários tecidos epiteliais, incluindo o estômago, as glândulas salivares e o pulmão. O linfoma MALT pulmonar tem origem no parênquima e/ou brônquios e é raro, representando 0.5-1% dos tumors primários do pulmão. Pode haver um atraso no tempo até ao diagnóstico porque a maioria dos pacientes é assintomática, sendo as investigações geralmente conduzidas pelo achado acidental de exames de imagem pulmonares alterados. Este linfoma tem um curso indolente e um bom prognóstico. Aqui expomos o caso de uma paciente assintomática com uma opacidade nodular que persistiu 5 meses após um episódio de pneumonia adquirida na comunidade. O diagnóstico final de linfoma MALT pulmonar foi obtido após uma investigação diagnóstica extensa.

Faecalibacterium prausnitzii is a major member of the Firmicutes phylum and one of the most abundant bacteria in the healthy human microbiota. F. prausnitzii depletion has been reported in several intestinal disorders, and more consistently in Crohn's disease (CD) patients. Despite its importance in human health, only few microbiological studies have been performed to isolate novel F. prausnitzii strains in order to better understand the biodiversity and physiological diversity of this beneficial commensal species. In this study, we described a protocol to isolate novel F. prausnitzii strains from feces of healthy volunteers as well as a deep molecular and metabolic characterization of these isolated strains. These F. prausnitzii strains were classified in two phylogroups and three clusters according to 16S rRNA sequences and results support that they would belong to two different genomospecies or genomovars as no genome sequencing has been performed in this work. Differences in enzymes production, antibiotic resistance and immunomodulatory properties were found to be strain-dependent. So far, all F. prausnitzii isolates share some characteristic such as (i) the lack of epithelial cells adhesion, plasmids, anti-microbial, and hemolytic activity and (ii) the presence of DNAse activity. Furthermore, Short Chain Fatty Acids (SCFA) production was assessed for the novel isolates as these products influence intestinal homeostasis. Indeed, the butyrate production has been correlated to the capacity to induce IL-10, an anti-inflammatory cytokine, in peripheral blood mononuclear cells (PBMC) but not to the ability to block IL-8 secretion in TNF-α-stimulated HT-29 cells, reinforcing the hypothesis of a complex anti-inflammatory pathway driven by F. prausnitzii. Altogether, our results suggest that some F. prausnitzii strains could represent good candidates as next-generation probiotic.

The extra-cellular microenvironment has a fundamental role in tumor growth and progression, strongly affecting the migration strategies adopted by single cancer cells during metastatic invasion. In this study, we use a novel microfluidic device to investigate the ability of mesenchymal and epithelial breast tumor cells to fluidize and migrate through narrowing microstructures upon chemoattractant stimulation. We compare the migration behavior of two mesenchymal breast cancer cell lines and one epithelial cell line, and find that the epithelial cells are able to migrate through the narrowest microconstrictions as the more invasive mesenchymal cells. In addition, we demonstrate that migration of epithelial cells through a highly compressive environment can occur in absence of a chemoattractive stimulus, thus evidencing that they are just as prone to react to mechanical cues as invasive cells.

La investigación tiene como objetivo conocer el uso que las agencias de comunicación publicitaria hacen de las redes sociales, los sectores que ya las utilizan y los sectores para los que pueden tener mayor potencia, y los fines y recursos estratégicos como canal comunicativo. Se han encuestado al 75% de las agencias socias de la Asociación Española de Comunicación Publicitaria (AEACP) utilizando técnicas del cuestionario y del análisis de contenido cualitativo. Los resultados confirman que el uso que hacen de las redes sociales se reflejan en las percepciones que tienen sobre las potencialidades del medio y que se las considera eficaces para desarrollar los objetivos de marca en los sectores que se dirigen a los jóvenes, las marcas relacionadas con el entretenimiento/ocio y para lanzar nuevos productos, desarrollar estrategias promocionales y fidelizar a los consumidores.

Excitotoxicity due to excessive activation of glutamate receptors is a primary mediator of cell death in acute and chronic neurological disorders, and NMDA-type glutamate receptors (NMDARs) are thought to be involved. NMDARs assemble from heteromeric combinations of GluN1, GluN2 and GluN3 subunits, yielding a variety of receptor subtypes that differ in biophysical properties, signaling, and synaptic targeting. Inclusion of inhibitory GluN3 subunits reduces Ca2+ influx via NMDAR channels and alters their synaptic targeting, thus modifying the two hallmarks of NMDARs that are critical for their roles on neuronal death and survival. Here we evaluated the neuroprotective potential of GluN3A subunits by analyzing the susceptibility to striatal excitotoxic damage of transgenic mice overexpressing GluN3A. We found that mild GluN3A overexpression protected susceptible striatal neurons from lesions induced by the neurotoxin 3-nitropropionic acid (3-NP), an inhibitor of mitochondrial complex II/succinate dehydrogenase. GluN3A-mediated neuroprotection was dose-dependent, and correlated with the levels of transgenic GluN3A expressed by two different mice strains. Neuroprotection was associated with a potent reduction of the activation of calpain, a Ca2+-dependent protease, which was measured as a decrease in 3-NP-induced fodrin and STEP cleavage in GluN3A transgenic mice relative to controls. We further show that transgenic GluN3A subunits incorporate into extrasynaptic compartments in mouse striatum, suggesting that reductions of toxic calpain activation might be linked to inhibition by GluN3A of pathological extrasynaptic NMDAR activity.

El presente artículo analiza con la finalidad de comprender mejor el ausentismo laboral: ¿Por qué no todos los individuos que enferman faltan al trabajo? ¿Qué tanto las ausencias laborales por enfermedad se relacionan realmente a la falta de salud? y ¿Qué tanto se relaciona el nivel de salud del trabajador con sus ausencias por causa diferente a enfermedad general? Asimismo, el artículo analiza en forma cualitativa cómo se mide el ausentismo de un trabajador cuando se trabaja a distancia (teletrabajo)

Femtosecond laser irradiation followed by chemical etching (FLICE) with hydrogen fluoride (HF) is an emerging technique for the fabrication of directly buried, three-dimensional microfluidic channels in silica. The procedure, as described in literature, consists of irradiating a silica slab followed by chemical etching using hydrogen fluoride. With aqueous HF the etching process is diffusion-limited and is self-terminating, leading to maximum microchannel lengths of about 1.5 mm, while the use of low-pressure gaseous HF etchant can quickly produce 3 mm long channels with an aspect ratio (Length/Diameter) higher than 25. By utilizing this methodology the aspect ratio is not constant, but depends on the length of the channel. When the microchannel is short the aspect ratio increases quickly until it reaches a maximum length at around 1400 µm. Thereafter the aspect ratio starts to decrease slowly. In this paper we present a variation of the low-pressure gaseous HF etching method, which is based on the dynamic displacement of the etchant. This method results in a 13% increase in the aspect ratio (L/D = 29) at the expense of a low etching speed (4 µm/min).

Microinjection moulding combined with the use of removable inserts is one of the most promising manufacturing processes for microfluidic devices, such as lab-on-chip, that have the potential to revolutionize the healthcare and diagnosis systems. In this work, we have designed, fabricated and tested a compact and disposable plastic optical stretcher. To produce the mould inserts, two micro manufacturing technologies have been used. Micro electro discharge machining (µEDM) was used to reproduce the inverse of the capillary tube connection characterized by elevated aspect ratio. The high accuracy of femtosecond laser micromachining (FLM) was exploited to manufacture the insert with perfectly aligned microfluidic channels and fibre slots, facilitating the final composition of the optical manipulation device. The optical stretcher operation was tested using microbeads and red blood cells solutions. The prototype presented in this work demonstrates the feasibility of this approach, which should guarantee real mass production of ready-to-use lab-on-chip devices.

Since the pioneering work of Ashkin and coworkers, back in 1970, optical manipulation gained an increasing interest among the scientific community. Indeed, the advantages and the possibilities of this technique are unsubtle, allowing for the manipulation of small particles with a broad spectrum of dimensions (nanometers to micrometers size), with no physical contact and without affecting the sample viability. Thus, optical manipulation rapidly found a large set of applications in different fields, such as cell biology, biophysics, and genetics. Moreover, large benefits followed the combination of optical manipulation and microfluidic channels, adding to optical manipulation the advantages of microfluidics, such as a continuous sample replacement and therefore high throughput and automatic sample processing. In this work, we will discuss the state of the art of these optofluidic devices, where optical manipulation is used in combination with microfluidic devices. We will distinguish on the optical method implemented and three main categories will be presented and explored: (i) a single highly focused beam used to manipulate the sample, (ii) one or more diverging beams imping on the sample, or (iii) evanescent wave based manipulation.

Se analizaron 16 muestras de maíz, 10 de maíz blanco nacional y 6 de maíz criollo blanco para determinar las especies de Fusarium presentes en los granos, en especial aquellas registradas como inductoras de pudriciones de mazorca y grano, principalmente las que producen micotoxinas. Las especies identificadas en orden del número de aislamientos obtenidos fueron: Fusarium oxysporum, F. subglutinans, F. moniliforme, F. graminearum, F. anthophilum, F. poae, F. tricinctum, F. sporotrichioides y F. proliferatum. Con excepción de F. oxysporum, F. tricinctum y F. anthophilum, las demás han sido registradas como inductoras de pudriciones de mazorca, grano y de tallo, y la mayoría son productoras de diversas micotoxinas, algunas de las cuales pueden ocasionar problemas a la salud humana y animal.Sixteen corn samples, 10 of national white corn and 6 of "criollo" white were analyzed to determine the Fusaria species present on the kernels, especially those reported as ear and kernel rot inducers, mainly those mycotoxin producers. The identified species, ordered in relation to the number of obtained isolates were F. oxysporum, F. subglutinans, F. moniliforme, F. graminearum, F. anthophilum, F. poae, F. tricinctum, F. sporotrichioides, and F. proliferatum. The species F. oxysporum, F. tricinctum, and F. anthophilum have not been reported causing ear or kernel rots. All of the others have been reported as ear and kernel rots inducers, as well as stalk rots. Besides, most of these species produce various mycotoxins, many of which are capable of causing health problems to humans and animals.

Alternative splicing is a key mechanism determinant for gene expression in metazoan. During alternative splicing, non-coding sequences are removed to generate different mature messenger RNAs due to a combination of sequence elements and cellular factors that contribute to splicing regulation. A different combination of splicing sites, exonic or intronic sequences, mutually exclusive exons or retained introns could be selected during alternative splicing to generate different mature mRNAs that could in turn produce distinct protein products. Alternative splicing is the main source of protein diversity responsible for 90% of human gene expression, and it has recently become a hallmark for cancer with a full potential as a prognostic and therapeutic tool. Currently, more than 15,000 alternative splicing events have been associated to different aspects of cancer biology, including cell proliferation and invasion, apoptosis resistance and susceptibility to different chemotherapeutic drugs. Here, we present well established and newly discovered splicing events that occur in different cancer-related genes, their modification by several approaches and the current status of key tools developed to target alternative splicing with diagnostic and therapeutic purposes.

Ultrafast laser microfabrication is a very powerful method for producing integrated devices in transparent materials [1].[...]

El consumo es un símbolo de integración porque se supone que cuando los nuevos residentes consumen se acercan al comportamiento de los nacionales. Por ello la publicidad sigue siendo una herramienta importante de inserción social a pesar de la crisis económica. Se hace un análisis detallado de las estrategias creativas y de la creatividad de los mensajes utilizados en las campañas dirigidas a nuevos residentes y se constata que se abusa de los estereotipos y que la mayoría de las campañas se concentran en tres sectores: comunicaciones, entidades financieras y alimentación y, en menor medida pero con un gran desarrollo en creatividad, en Codesarrollo tal y como se observa en el Festival Universitario de Publicidad Social Solidar&dar de la Universidad Rey Juan Carlos dirigido por Isidoro Arroyo.The consumption is a symbol to integrate inmigrants because inmigrants do the same things that natives. Advertisement continues been an important tool to integrate inmigrants. This article analices creative strategy and messages analysis in campaign manages to inmigrants. We consider that they exceeds of stereotypes. The most of the campaign are about communication, banks and food. They are about Co-development too as we could see in Social Universitary Advertisement Festival Solidar&dar in Rey Juan Carlos University whose chairman is Isidoro Arroyo.

Flash photolysis of caged compounds is one of the most powerful approaches to investigate the dynamic response of living cells. Monolithically integrated devices suitable for optical uncaging are in great demand since they greatly simplify the experiments and allow their automation. Here we demonstrate the fabrication of an integrated bio-photonic device for the optical release of caged compounds. Such a device is fabricated using femtosecond laser micromachining of a glass substrate. More in detail, femtosecond lasers are used both to cut the substrate in order to create a pit for cell growth and to inscribe optical waveguides for spatially selective uncaging of the compounds present in the culture medium. The operation of this monolithic bio-photonic device is tested using both free and caged fluorescent compounds to probe its capability of multipoint release and optical sensing. Application of this device to the study of neuronal network activity can be envisaged.

Este artículo presenta un estudio de caso sobre la comunicación que realizan, a través de Internet, las organizaciones no gubernamentales de desarrollo (ongd), especialmente aquellas que han tenido un reconocimiento administrativo a su gestión por la Agencia Española de Cooperación Internacional (aeci). Los resultados confirman el conocimiento que tienen las ongd de la necesidad de estar en la Red, de las posibilidades de mejora que se abren en el desarrollo de sus sitios web diseño, interactividad, accesibilidad, etc.- así como de la necesidad de seguir investigando en este ámbito para desarrollar estrategias de comunicación web acordes con las necesidades de las entidades sin ánimo de lucro.

Hemos revisado de forma retrospectiva 18 casos de tumor filodes, diagnosticados histopatológi-camente durante un período de 6 años, recogiendo los datos clínicos, radiológicos y evolutivos. Todos los casos se presentaron como nodulo palpable, fibroelástico e indoloro excepto uno que lo hizo como mastitis sobre tumoración. En mamografía mostraron densidad homogénea, bordes relativamente bien definidos, sin calcificaciones en su interior. Ultraso-nográficamente aparecieron como masas sólidas, heterogéneas, con refuerzo posterior y algunas de ellas con áreas quísticas características. En todos los casos se realizó tratamiento quirúrgico y en 15 pacientes se hizo punción aspirativa con aguja fina (PAAF), previo a la cirugía. Los datos evolutivos en relación a recidiva local y metástasis son similares a los descritos en la bibliografía. Recomendamos estrecha vigilancia clínico-radiológica debido a la historia natural del tumor y su alto número de recidivasWe have revised in retrospective study, 18 cases of phyllodes breast tumor diagnosed histolo-gically over a 6 year-period, gathering the clinical, radiological and evolutionary data. All the cases appeared as palpable, elastic and painless nodules. Mammography showed homogeneous density, relatively well defined borders, without internal calcifications. Ultrasonographically they appeared as solid masses, heterogeneous, with posterior enhancement and, some of them with characteristic cystic areas. Fine needle aspiration biopsy was made in 15 patients, before the surgical treatment that was carried out in all the cases. The information related to local relapse and metastasis is similar to those described in the bibliography. We recommend close clinical-radiological surveillance, due to the natural history of the tumour and their high number of relapses

A entrada na Universidade para muitos jovens é marcada por uma fase de transição caracterizada por diversas mudanças que interferem em seu desenvolvimento psicossocial, além do profissional. O presente trabalho teve como objetivo a análise dos fatores que influenciaram na adaptação do aluno, egresso do ensino médio, à vida universitária, bem como a identificação das mudanças comportamentais ocorridas em função desse processo de transição acadêmica. O estudo foi realizado através de entrevistas semi-estrutadas com 8 estudantes pertencentes a diferentes cursos. Os dados foram analisados através da análise de conteúdo e categorizados em dois segmentos: Fatores que Influenciaram na Adaptação e Mudanças Comportamentais. Os resultados indicaram o destaque de sete categorias para o primeiro bloco – Infraestrutura; Informação sobre a Universidade e o curso; Fatores Externo à Universidade; Aspectos Organizacionais; Material de Estudo; Vínculos e Professores – e quatro categorias para o segundo bloco - Aspectos pessoais; Organização pessoal para o estudo; Tempo e Sintomas. O estudo contribui para pontuar as dificuldades encontradas pelos alunos ingressantes no contexto da universidade e proporcionou melhor compreensão sobre a transição acadêmica.

We have introduced a new hybrid fabrication method for lab-on-a-chip devices through the combination of femtosecond laser micromachining and removable insert micro-injection molding. This method is particularly suited for the fast prototyping of new devices, while maintaining a competitive low cost. To demonstrate the effectiveness of our approach, we designed, fabricated, and tested a completely integrated flow cytometer coupled to a portable media device. The system operation was tested with fluorescent plastic micro-bead solutions ranging from 100 beads/μL to 500 beads/μL. We demonstrated that this hybrid lab-on-a-chip fabrication technology is suitable for producing low-cost and portable biological microsystems and for effectively bridging the gap between new device concepts and their mass production.

The 3' untranslated region (3'UTR) of human astroviruses (HAstV) consists of two hairpin structures (helix I and II) joined by a linker harboring a conserved PTB/hnRNP1 binding site. The identification and characterization of cellular proteins that interact with the 3'UTR of HAstV-8 virus will help to uncover cellular requirements for viral functions. To this end, mobility shift assays and UV cross-linking were performed with uninfected and HAstV-8-infected cell extracts and HAstV-8 3'UTR probes. Two RNA-protein complexes (CI and CII) were recruited into the 3'UTR. Complex CII formation was compromised with cold homologous RNA, and seven proteins of 35, 40, 45, 50, 52, 57/60 and 75 kDa were cross-linked to the 3'UTR. Supermobility shift assays indicated that PTB/hnRNP1 is part of this complex, and 3'UTR-crosslinked PTB/hnRNP1 was immunoprecipitated from HAstV-8 infected cell-membrane extracts. Also, immunofluorescence analyses revealed that PTB/hnRNP1 is distributed in the nucleus and cytoplasm of uninfected cells, but it is mainly localized perinuclearly in the cytoplasm of HAstV-8 infected cells. Furthermore, the minimal 3'UTR sequences recognized by recombinant PTB are those conforming helix I, and an intact PTB/hnRNP1-binding site. Finally, small interfering RNA-mediated PTB/hnRNP1 silencing reduced synthesis viral genome and virus yield in CaCo2 cells, suggesting that PTB/hnRNP1 is required for HAstV replication. In conclusion, PTB/hnRNP1 binds to the 3'UTR HAstV-8 and is required or participates in viral replication.

The cobas 4800 BRAF V600 Mutation Test is a CE-marked and FDA-approved in vitro diagnostic assay used to select patients with metastatic melanoma for treatment with the selective BRAF inhibitor vemurafenib. We describe the pre-approval validation of this test in two external laboratories.Melanoma specimens were tested for BRAF V600 mutations at two laboratories with the: cobas BRAF Mutation Test; ABI BRAF test; and bidirectional direct sequencing. Positive (PPA) and negative (NPA) percent agreements were determined between the cobas test and the other assays. Specimens with discordant results were tested with massively parallel pyrosequencing (454). DNA blends with 5% mutant alleles were tested to assess detection rates.Invalid results were observed in 8/116 specimens (6·9%) with Sanger, 10/116 (8·6%) with ABI BRAF, and 0/232 (0%) with the cobas BRAF test. PPA was 97·7% for V600E mutation for the cobas BRAF test and Sanger, and NPA was 95·3%. For the cobas BRAF test and ABI BRAF, PPA was 71·9% and NPA 83·7%. For 16 cobas BRAF test-negative/ABI BRAF-positive specimens, 454 sequencing detected no codon 600 mutations in 12 and variant codon 600 mutations in four. For eight cobas BRAF test-positive/ABI BRAF-negative specimens, four were V600E and four V600K by 454 sequencing. Detection rates for 5% mutation blends were 100% for the cobas BRAF test, 33% for Sanger, and 21% for the ABI BRAF. Reproducibility of the cobas BRAF test was 111/116 (96%) between the two sites.It is feasible to evaluate potential companion diagnostic tests in external laboratories simultaneously to the pivotal clinical trial validation. The health authority approved assay had substantially better performance characteristics than the two other methods. The overall success of the cobas BRAF test is a proof of concept for future biomarker development.

Assessing the distribution of 16S rRNA gene sequences within a biological sample represents the current state-of-the-art for determination of human gut microbiota composition. Advances in dissecting the microbial biodiversity of this ecosystem have very much been dependent on the development of novel high-throughput DNA sequencing technologies, like the Ion Torrent. However, the precise representation of this bacterial community may be affected by the protocols used for DNA extraction as well as by the PCR primers employed in the amplification reaction. Here, we describe an optimized protocol for 16S rRNA gene-based profiling of the fecal microbiota.

El artículo presenta al teletrabajo como la última trasformación del trabajo a través de la historia del pensamiento económico. Se analiza desde diversos aspectos tales como su concepto y propuesta de clasificación, el derecho laboral, la motivación y el perfil de los teletrabajadores así como sus áreas de aplicación. El teletrabajo es una tendencia laboral actual que está trasformando la idea de que el ser humano debe salir de casa y trasladarse físicamente a un lugar para trabajar, hoy día el trabajo puede realizarse en cualquier lugar en el que se cuente con computadora e internet, este hecho requiere analizar los cambios en materia económica, administrativa y jurídica que conlleva.

The objective of this study is to compare two EGFR testing methodologies (a commercial real-time PCR kit and a specific EGFR mutant immunohistochemistry), with direct sequencing and to investigate the limit of detection (LOD) of both PCR-based methods. We identified EGFR mutations in 21 (16%) of the 136 tumours analyzed by direct sequencing. Interestingly, the Therascreen EGFR Mutation Test kit was able to characterize as wild-type one tumour that could not be analyzed by direct sequencing of the PCR product. We then compared the LOD of the kit and that of direct sequencing using the available mutant tumours. The kit was able to detect the presence of a mutation in a 1% dilution of the total DNA in nine of the 18 tumours (50%), which tested positive with the real-time quantitative PCR method. In all cases, EGFR mutation was identified at a dilution of 5%. Where the mutant DNA represented 30% of the total DNA, sequencing was able to detect mutations in 12 out of 19 cases (63%). Additional experiments with genetically defined standards (EGFR ΔE746-A750/+ and EGFR L858R/+) yielded similar results. Immunohistochemistry (IHC) staining with exon 19-specific antibody was seen in eight out of nine cases with E746-A750del detected by direct sequencing. Neither of the two tumours with complex deletions were positive. Of the five L858R-mutated tumours detected by the PCR methods, only two were positive for the exon 21-specific antibody. The specificity was 100% for both antibodies. The LOD of the real-time PCR method was lower than that of direct sequencing. The mutation specific IHC produced excellent specificity.

Las miradas que se lanzan a los países en vías de desarrollo desde los que no lo son siguen siendo miopes y estereotipadas. Se fomenta, además, a través de las representaciones que se ofrecen de ellos en los medios de comunicación de masas. La imagen que llega a los países del conocido como primer mundo pasa por el tamiz de la espectacularidad de las guerras, los desastres naturales y lo sugerente de la publicidad. Las ONG, las instituciones que más conocen estos países, sus culturas y sus gentes, aún deben trabajar para crear una percepción más cercana y certera de las realidades del Tercer Mundo. Las imágenes que utilizan en la actualidad han evolucionado y se han regulado por códigos deontológicos, pero no consiguen eliminar la imagen de estas sociedades como alejadas, dependientes e infantilizadas.

hnRNP A/B proteins modulate the alternative splicing of several mammalian and viral pre-mRNAs, and are typically viewed as proteins that enforce the activity of splicing silencers. Here we show that intronic hnRNP A/B-binding sites (ABS) can stimulate the in vitro splicing of pre-mRNAs containing artificially enlarged introns. Stimulation of in vitro splicing could also be obtained by providing intronic ABS in trans through the use of antisense oligonucleotides containing a non-hybridizing ABS-carrying tail. ABS-tailed oligonucleotides also improved the in vivo inclusion of an alternative exon flanked by an enlarged intron. Notably, binding sites for hnRNP F/H proteins (FBS) replicate the activity of ABS by improving the splicing of an enlarged intron and by modulating 5' splice-site selection. One hypothesis formulated to explain these effects is that bound hnRNP proteins self-interact to bring in closer proximity the external pair of splice sites. Consistent with this model, positioning FBS or ABS at both ends of an intron was required to stimulate splicing of some pre-mRNAs. In addition, a computational analysis of the configuration of putative FBS and ABS located at the ends of introns supports the view that these motifs have evolved to support cooperative interactions. Our results document a positive role for the hnRNP A/B and hnRNP F/H proteins in generic splicing, and suggest that these proteins may modulate the conformation of mammalian pre-mRNAs.

Acuabio 1 is a mixture of proteins and essential amino acids that exerts a stimulating effect in the development of aquatic organisms. It synchronizes and accelerates fish and shrimp larvae growth on early developmental stages, as well as increases the development and size of these animals. In this work, the stimulating effect of Acuabio 1 on the immune system of Litopennaeus vannamei shrimps was demonstrated by obtaining (5.4 x 10(6) hemocytes/mL) greater than the control group (4.3 x 10(6) hemocytes /mL). The hemolymph of treated animals showed higher lectin titers (16) than the control group (8). Animals treated with Acuabio 1 increased their phenoloxidase and peroxidase activities, indicating an enhanced oxidative defense. Furthermore, Acuabio 1 decreased the number of Vibrio spp. and Pseudomonas spp. colony forming units in the culture medium, suggesting a possible prebiotic function. All these properties make Acuabio 1 very attractive to improve the production, survival rate and quality of shrimp larvae.

Hemos revisado de forma retrospectiva 18 casos de tumor filodes, diagnosticados histopatológi-camente durante un período de 6 años, recogiendo los datos clínicos, radiológicos y evolutivos. Todos los casos se presentaron como nodulo palpable, fibroelástico e indoloro excepto uno que lo hizo como mastitis sobre tumoración. En mamografía mostraron densidad homogénea, bordes relativamente bien definidos, sin calcificaciones en su interior. Ultraso-nográficamente aparecieron como masas sólidas, heterogéneas, con refuerzo posterior y algunas de ellas con áreas quísticas características. En todos los casos se realizó tratamiento quirúrgico y en 15 pacientes se hizo punción aspirativa con aguja fina (PAAF), previo a la cirugía. Los datos evolutivos en relación a recidiva local y metástasis son similares a los descritos en la bibliografía. Recomendamos estrecha vigilancia clínico-radiológica debido a la historia natural del tumor y su alto número de recidivas

In teleost fish, secretion of the growth hormone (GH) is regulated by several hypothalamic factors that are influenced by the physiological state of the animal. GH in fish is involved in many physiological processes that are not overtly growth-related such as: saltwater osmoregulation, antifreeze protein synthesis, and the regulation of sexual maturation and immune functions. This study was conducted to characterize a decapeptide A233 (GKFDLSPEHQ) designed by molecular modeling to evaluate its function as a GH secretagogue (GHS). In pituitary cell culture, the peptide A233 induces GH secretion and it is also able to increase superoxide production in tilapia head-kidney leukocyte cultures. This effect is blocked by preincubation with the GHS receptor antagonist [D-Lys3]-GHRP6. Immunoneutralization of GH by addition of anti-tilapia GH monoclonal antibody blocked the stimulatory effect of A233 on superoxide production. These experiments suggest a GH-mediated mechanism for the action of A233. The in vivo biological action of the decapeptide was also demonstrated for growth stimulation in goldfish and tilapia larvae (p < 0.001). Superoxide dismutase levels, antiprotease activity, and lectin titer were enhanced in tilapia larvae treated with this novel molecule. The decapeptide A233 is able to function as GHS in teleosts and enhance parameters of the innate immune system in the fish larvae. This study won the Annual Award of the Academy of Sciences of Cuba in 2012.