Nuclear/Cytoplasmic Fractionation of Proteins from Caenorhabditis elegans

C. elegans is widely used to investigate biological processes related to health and disease. To study protein localization, fluorescently-tagged proteins can be used in vivo or immunohistochemistry can be performed in whole worms. Here, we describe a technique to localize a protein of interest at a subcellular level in C. elegans lysates, which can give insight into the location, function and/or toxicity of proteinsNational Institutes of Health National Centre for Research Resources (NIH)European Research Council (ERC)USANIH National Center for Research Resources (NCRR)Japan National BioResource Projec

Filter Retardation Assay for Detecting and Quantifying Polyglutamine Aggregates Using Caenorhabditis elegans Lysates

Protein aggregation is a hallmark of several neurodegenerative diseases and is associated with impaired protein homeostasis. This imbalance is caused by the loss of the protein's native conformation, which ultimately results in its aggregation or abnormal localization within the cell. Using a C. elegans model of polyglutamine diseases, we describe in detail the filter retardation assay, a method that captures protein aggregates in a cellulose acetate membrane and allows its detection and quantification by immunoblotting

Evidence of learning on the insertion and care of peripheral venous catheters in nursing students: A mixed study

Objectives: 1) To assess nursing students' evidence-based knowledge on the use of PVCs, and 2) to examine the perception of learning and teaching strategies aimed at this skill. Background: Insertion and care of Peripheral Venous Catheters (PVCs) are essential skills in undergraduate nursing education. Appropriate knowledge of this skill is crucial to improve clinical practice and patient safety. Therefore, training becomes an enabler for safe practice. Design: A multi-centre convergent parallel mixed-methods. Setting and participants: A total of 675 second-, third- and fourth-year nursing students from 3 nursing schools took part in the study. Methods: Quantitative data collection used a validated 15-question survey on knowledge of PVC management, and a descriptive and inferential analysis was carried out. Qualitative data were collected via a questionnaire consisting of 4 open-ended questions assessing knowledge, teaching methodologies and scenarios, and points for improvement. Results: Most participants were female (74.04%), with a mean age of 22.45 (SD = 4.65), who had no experience in the health field (61.8%). They obtained a mean knowledge score of 7.27 (SD = 2.64) out of 15. The students who obtained higher scores had a mean professional experience of 7.96, SD = 2.66 (p 0.000) and were in their final year, with a mean of 8.59, SD = 2.56, (p 0.000). On the other hand, the students assessed their knowledge as basic but improving year by year. They also identified a need to apply more active and experiential methodologies that would allow for reflection. Conclusion: Level of educational level and experience is associated with increased knowledge. In order to improve knowledge, changes must be made in the training process to incorporate methodologies such as simulation and online training. There is a need to develop programmes that favour the alignment of theory with clinical practice

Neuronal accumulation of unrepaired DNA in a novel specific chromatin domain: structural, molecular and transcriptional characterization

There is growing evidence that defective DNA repair in neurons with accumulation of DNA lesions and loss of genome integrity underlies aging and many neurodegenerative disorders. An important challenge is to understand how neurons can tolerate the accumulation of persistent DNA lesions without triggering the apoptotic pathway. Here we study the impact of the accumulation of unrepaired DNA on the chromatin architecture, kinetics of the DNA damage response and transcriptional activity in rat sensory ganglion neurons exposed to 1-to-3 doses of ionizing radiation (IR). In particular, we have characterized the structural, molecular and transcriptional compartmentalization of unrepaired DNA in persistent DNA damaged foci (PDDF). IR induced the formation of numerous transient foci, which repaired DNA within the 24 h post-IR, and a 1-to-3 PDDF. The latter concentrate DNA damage signaling and repair factors, including ?H2AX, pATM, WRAP53 and 53BP1. The number and size of PDDF was dependent on the doses of IR administered. The proportion of neurons carrying PDDF decreased over time of post-IR, indicating that a slow DNA repair occurs in some foci. The fine structure of PDDF consisted of a loose network of unfolded 30 nm chromatin fiber intermediates, which may provide a structural scaffold accessible for DNA repair factors. Furthermore, the transcription assay demonstrated that PDDF are transcriptionally silent, although transcription occurred in flanking euchromatin. Therefore, the expression of ?H2AX can be used as a reliable marker of gene silencing in DNA damaged neurons. Moreover, PDDF were located in repressive nuclear environments, preferentially in the perinucleolar domain where they were frequently associated with Cajal bodies or heterochromatin clumps forming a structural triad. We propose that the sequestration of unrepaired DNA in discrete PDDF and the transcriptional silencing can be essential to preserve genome stability and prevent the synthesis of aberrant mRNA and protein products encoded by damaged genes

The state of the Martian climate

60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

Persistent accumulation of unrepaired DNA damage in rat cortical neurons: nuclear organization and ChIP-seq analysis of damaged DNA

Neurons are highly vulnerable to DNA damage induced by genotoxic agents such as topoisomerase activity, oxidative stress, ionizing radiation (IR) and chemotherapeutic drugs. To avert the detrimental effects of DNA lesions in genome stability, transcription and apoptosis, neurons activate robust DNA repair mechanisms. However, defective DNA repair with accumulation of unrepaired DNA are at the basis of brain ageing and several neurodegenerative diseases. Understanding the mechanisms by which neurons tolerate DNA damage accumulation as well as defining the genomic regions that are more vulnerable to DNA damage or refractory to DNA repair and therefore constitute potential targets in neurodegenerative diseases are essential issues in the field. In this work we investigated the nuclear topography and organization together with the genome-wide distribution of unrepaired DNA in rat cortical neurons 15 days upon IR. About 5% of non-irradiated and 55% of irradiated cells accumulate unrepaired DNA within persistent DNA damage foci (PDDF) of chromatin. These PDDF are featured by persistent activation of DNA damage/repair signaling, lack of transcription and localization in repressive nuclear microenvironments. Interestingly, the chromatin insulator CTCF is concentrated at the PDDF boundaries, likely contributing to isolate unrepaired DNA from intact transcriptionally active chromatin. By confining damaged DNA, PDDF would help preserving genomic integrity and preventing the production of aberrant proteins encoded by damaged genes.ChIP-seq analysis of genome-wide ?H2AX distribution revealed a number of genomic regions enriched in ?H2AX signal in IR-treated cortical neurons. Some of these regions are in close proximity to genes encoding essential proteins for neuronal functions and human neurodegenerative disorders such as epm2a (Lafora disease), serpini1 (familial encephalopathy with neuroserpin inclusion bodies) and il1rpl1 (mental retardation, X-linked 21). Persistent ?H2AX signal close to those regions suggests that nearby genes could be either more vulnerable to DNA damage or more refractory to DNA repair.This work was supported by the following grants: “Dirección General de Investigación” (BFU2014–54754-P) and “Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas” (CIBERNED; CB06/05/0037) Spain

Jardins per a la salut

Facultat de Farmàcia, Universitat de Barcelona. Ensenyament: Grau de Farmàcia. Assignatura: Botànica farmacèutica. Curs: 2014-2015. Coordinadors: Joan Simon, Cèsar Blanché i Maria Bosch.Els materials que aquí es presenten són el recull de les fitxes botàniques de 128 espècies presents en el Jardí Ferran Soldevila de l’Edifici Històric de la UB. Els treballs han estat realitzats manera individual per part dels estudiants dels grups M-3 i T-1 de l’assignatura Botànica Farmacèutica durant els mesos de febrer a maig del curs 2014-15 com a resultat final del Projecte d’Innovació Docent «Jardins per a la salut: aprenentatge servei a Botànica farmacèutica» (codi 2014PID-UB/054). Tots els treballs s’han dut a terme a través de la plataforma de GoogleDocs i han estat tutoritzats pels professors de l’assignatura. L’objectiu principal de l’activitat ha estat fomentar l’aprenentatge autònom i col·laboratiu en Botànica farmacèutica. També s’ha pretès motivar els estudiants a través del retorn de part del seu esforç a la societat a través d’una experiència d’Aprenentatge-Servei, deixant disponible finalment el treball dels estudiants per a poder ser consultable a través d’una Web pública amb la possibilitat de poder-ho fer in-situ en el propi jardí mitjançant codis QR amb un smartphone

Antibacterial Residue Excretion via Urine as an Indicator for Therapeutical Treatment Choice and Farm Waste Treatment

Many of the infectious diseases that affect livestock have bacteria as etiological agents. Thus, therapy is based on antimicrobials that leave the animal’s tissues mainly via urine, reaching the environment through slurry and waste water. Once there, antimicrobial residues may lead to antibacterial resistance as well as toxicity for plants, animals, or humans. Hence, the objective was to describe the rate of antimicrobial excretion in urine in order to select the most appropriate molecule while reducing harmful effects. Thus, 62 pigs were treated with sulfamethoxypyridazine, oxytetracycline, and enrofloxacin. Urine was collected through the withdrawal period and analysed via LC-MS/MS. Oxytetracycline had the slowest rate of degradation (a half-life time of 4.18 days) and the most extended elimination period in urine (over 2 months), followed by enrofloxacin (a half-life time of 1.48 days, total urine elimination in ca. 3 weeks) and sulfamethoxypyridazine (a half-life time of 0.49 days, total urine elimination in ca. 1 week). Bacterial sensitivity and recommendations for responsible use are limiting when selecting the treatment. Nevertheless, with similar effectiveness, sulfamethoxypyridazine would be the choice, as waste treatment would only need to be implemented for 1 week after treatment. Thus, more in-depth knowledge regarding antibacterial elimination would improve resource management, while protecting animals and consumers’ health

Relación del bajo peso al nacer con el desarrollo psicomotor del niño de Puriscal

Artículo científico -- Universidad de Costa Rica, Instituto de Investigaciones en Salud. 1985This is a study of psycho-motor development of 84 eighteen months old children, living in the districts of Barbacoas, Grifo Alto and Candelarita in the canton of Puriscal. Gesell and Casati-Lezine tests were employed to study motricity, adaptation, languaje and several other areas of psycho-motor development. A total developmental index (D.I) was obtained and related with children's gestational age and weight at birth. Preterrn babies showed lowered D. I. than babies born at term (p=0,003). Direct relationship between weight at birth and D. I. (r=0,39, p=0,001). D. I. was even lower when low birth weight and prematurity were considered togetherSe realizó un estudio del desarrollo psicomotor en 84 niños de los distritos de Barbacoas, Grifo Alto y Candelarita de Puriscal, a los 18 meses de edad. El estudio comprendió las áreas de motricidad, adaptación, lenguaje, personal- social, utilización de recursos, búsqueda del objeto y exploración, evaluadas por media de pruebas de desarrollo psicomotor de Gesell y Casati-Lezine. Luego de evaluar las áreas se obtuvo un índice de desarrollo (l.D.) global que fue relacionado con la edad gestacional y el peso al nacer de los niños. Los niños de pre término mostraron I.D. inferiores en relación con los nacidos a término (13=0,003). Además, se encontró una relación directa entre el peso al nacer y el índice de desarrollo, pues cuanto mes bajo era el peso, más bajo fue el Índice (r=0,39, p=0,001). Al combinarse el bajo peso al nacer con la prematuridad el I.D. fue aún más bajo.Universsidad de Costa Rica, Instituto de Investigaciones en SaludUniversidad de Costa Rica, Escuela de PsicologíaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA