Inflammation protein quantification by multiple reaction monitoring mass spectrometry in lipopolysaccharide-stimulated THP-1 cells

Rationale: Inflammation is a cascade of events mediated by a cytokine network triggering the cellular response. In order to monitor the modulation of the crucial inflammatory proteins, e.g., Tumour Necrosis Factor-α (TNF-α), Interferon-γ (INF-γ), Interleukin-8 (IL-8) and Interleukin-10 (IL-10), upon stimulation with endotoxins, differentiated and undifferentiated THP-1 cells were treated with lipopolysaccharides (LPSs) from E. coli, key cell wall components of Gram-negative bacteria. Methods: The multiple reaction monitoring mass spectrometry (MRM-MS) method was optimized by using the standard proteins to be quantified, in order to construct external calibration curves and define the analytical parameters. The developed method was used to quantify the above-mentioned inflammatory proteins in THP-1 differentiated cells upon stimulation with LPSs with high accuracy, sensitivity, and robustness. Results: The analysis of such proteins in MRM mode allowed the kinetics of stimulation along the time up to 24 h to be followed and the MS results were found to be comparable with those obtained by Western-blotting. A significant increase in TNF-α release triggered a cascade mechanism leading to the production of INF-γ and IL-8. IL-10, instead, was found to be constant throughout the process. Conclusions: The developed MRM-MS method allowed the quantification of TNF-α, INF-γ, IL-8 and IL-10 along a time-course from 2 to 24 h. Hence, a trace of the kinetics of the inflammatory response in THP-1 cells upon stimulation with E. coli LPSs was obtained. Finally, the extensibility of the developed MRM method to serum samples and other matrices demonstrated the versatility of the approach and the possibility to quantify multiple target proteins in different biological samples by using a few microliters in a single analysis

Tracking defect-induced ferromagnetism in GaN:Gd

We report on the magnetic properties of GaN:Gd layers grown by molecular beam epitaxy (MBE). A poor reproducibility with respect to the magnetic properties is found in these samples. Our results show strong indications that defects with a concentration of the order of 10^19 cm^-3 might play an important role for the magnetic properties. Positron annihilation spectroscopy does not support the suggested connection between the ferromagnetism and the Ga vacancy in GaN:Gd. Oxygen co-doping of GaN:Gd promotes ferromagnetism at room temperature and points to a role of oxygen for mediating ferromagnetic interactions in Gd doped GaN

Behavioral approaches to weight control: A review of current research

Weight management is a salient issue for women. Studies of behavioral, pharmacological and surgical interventions indicate that women comprise the majority of patients presenting for weight-loss treatment. In this review we discuss the health impact of obesity for women, review behavioral treatments for adult overweight and obesity, and address topics of particular relevance for women, including concerns that weight-loss treatment may precipitate the development of eating pathology, as well as time periods of high risk for weight gain such as pregnancy and menopause

The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins

: Antimicrobial peptides (AMPs) are a unique and diverse group of molecules endowed with a broad spectrum of antibiotics properties that are being considered as new alternative therapeutic agents. Most of these peptides are membrane-active molecules, killing bacteria by membrane disruption. However, recently an increasing number of AMPs was shown to enter bacterial cells and target intracellular processes fundamental for bacterial life. In this paper we investigated the mechanism of action of Maganin-2 (Mag-2), a well-known antimicrobial peptide isolated from the African clawed frog Xenopus laevis, by functional proteomic approaches. Several proteins belonging to E. coli macromolecular membrane complexes were identified as Mag-2 putative interactors. Among these, we focused our attention on BamA a membrane protein belonging to the BAM complex responsible for the folding and insertion of nascent β-barrel Outer Membrane Proteins (OMPs) in the outer membrane. In silico predictions by molecular modelling, in vitro fluorescence binding and Light Scattering experiments carried out using a recombinant form of BamA confirmed the formation of a stable Mag-2/BamA complex and indicated a high affinity of the peptide for BamA. Functional implications of this interactions were investigated by two alternative and complementary approaches. The amount of outer membrane proteins OmpA and OmpF produced in E. coli following Mag-2 incubation were evaluated by both western blot analysis and quantitative tandem mass spectrometry in Multiple Reaction Monitoring scan mode. In both experiments a gradual decrease in outer membrane proteins production with time was observed as a consequence of Mag-2 treatment. These results suggested BamA as a possible good target for the rational design of new antibiotics since this protein is responsible for a crucial biological event of bacterial life and is absent in humans

Propuesta de concientización sobre la importancia de la preservación de la fauna local de la vereda Santa Bárbara-El Playón

Con la intención de generar conciencia en la comunidad de la localidad de la vereda Santa Bárbara, del municipio de El Playón, del departamento de Santander, se ha propuesto esta intervención pedagógica para trabajar con los estudiantes de básica secundaria de la escuela rural Santa Bárbara. El proyecto consiste en el diseño de una propuesta de concientización sobre la biodiversidad del entorno, mediante instrumentos como la encuesta y la observación participante, con la finalidad de que los estudiantes puedan interactuar de forma más directa con la fauna silvestre local y, de esta manera, se fomente un sentido de pertenencia hacia el ambiente que los rodea, y se promuevan actitudes más responsables con los animales. La presente propuesta plantea una solución potencial relacionada con el estudio y el cuidado de la fauna silvestre local y se esperan cambios cada vez más grandes y significativos, partiendo desde la escuela, con el fin de que no sólo se beneficien los estudiantes de la institución educativa, sino también sus familias y demás habitantes de la vereda Santa Bárbara.With the intention of generating awareness in the community of the village of Santa Bárbara, in the municipality of El Playón, in the department of Santander, this pedagogical intervention has been proposed to work with high school students of the Santa Bárbara rural school. The project consists of designing a proposal to raise awareness about the biodiversity of the environment, using instruments such as surveys and participant observation, so that students can interact more directly with the local wildlife and, in this way, foster a sense of belonging to the environment that surrounds them, and promote more responsible attitudes towards animals. The present proposal proposes a potential solution related to the study and care of the local wildlife and it is expected to bring about greater and more significant changes, starting from the school, in order to benefit not only the students of the educational institution, but also their families and other inhabitants of the Santa Bárbara village

The Value of Removing Daily Obstacles via Everyday Problem-Solving Theory: Developing an Applied Novel Procedure to Increase Self-Efficacy for Exercise

The objective of the study was to develop a novel procedure to increase self-efficacy for exercise. Gains in one’s ability to resolve day-to-day obstacles for entering an exercise routine were expected to cause an increase in self-efficacy for exercise. Fifty-five sedentary participants (did not exercise regularly for at least 4 months prior to the study) who expressed an intention to exercise in the near future were selected for the study. Participants were randomly assigned to one of three conditions: (1) an Experimental Group in which they received a problem-solving training session to learn new strategies for solving day-to-day obstacles that interfere with exercise, (2) a Control Group with Problem-Solving Training which received a problem-solving training session focused on a typical day-to-day problem unrelated to exercise, or (3) a Control Group which did not receive any problem-solving training. Assessment of obstacles to exercise and perceived self-efficacy for exercise were conducted at baseline; perceived self-efficacy for exercise was reassessed post-intervention (1 week later). No differences in perceived challenges posed by obstacles to exercise or self-efficacy for exercise were observed across groups at baseline. The Experimental Group reported greater improvement in self-efficacy for exercise compared to the Control Group with Training and the Control Group. Results of this study suggest that a novel procedure that focuses on removing obstacles to intended planned fitness activities is effective in increasing self-efficacy to engage in exercise among sedentary adults. Implications of these findings for use in applied settings and treatment studies are discussed

Cuidados Paliativos no Âmbito do SUS

Os cuidados paliativos são uma necessidade urgente e humanitária de natureza global para as pessoas com câncer ou doenças ameaçadoras da vida. Recomenda-se que devam ser ofertados a partir do diagnóstico, associados ou não à terapia curativa, priorizando o manejo dos sintomas físicos, psíquicos, sócio familiares e o apoio espiritual. Para tal, deve ser adaptado às necessidades dos pacientes e suas famílias, acompanhando a progressão da doença até sua eventual fase final. E pode continuar após a morte do paciente através do apoio às famílias em seu processo de luto. A proposta de construir uma politica de cuidados paliativos no âmbito do SUS deve ter uma abordagem que promove a qualidade de vida de pacientes e seus familiares, que enfrentam doenças que ameacem a continuidade da vida, por meio da prevenção e do alívio do sofrimento. Versando na identificação precoce, avaliação e tratamento impecável da dor e outros problemas de natureza física, psíquica, social e espiritual. Os cuidados paliativos ocorrem em diversos níveis de atenção, sendo imprescindível a articulação entre os diversos serviços e as ações realizadas de forma que, independentemente do local onde sejam prestados, estejam voltados ao alcance de um objetivo comum, a melhoria da qualidade de vida e a manutenção da dignidade dos cidadãos brasileiros ao longo do processo de finitude. É necessário que haja articulação da atenção básica com os serviços domiciliares, ambulatoriais especializados e hospitalares, favorecendo o diagnóstico rápido das necessidades dos pacientes e a oferta de tratamento adequad

Fertility preservation in gynaecologic cancers

Due to substantial improvement in the diagnosis and treatment of gynaecologic cancers, a better understanding of patient care needs to be revised. We reviewed the literature related to fertility preservation strategies in gynaecological cancer and discussed current general management approaches. New technical modalities and patients’ own desire for motherhood should be integral and paramount in the clinical evaluation to significantly contribute to preserving fertility in those women diagnosed with gynaecologic cancers during the reproductive years