Differential Internalization of Amphotericin B - Conjugated Nanoparticles in Human Cells and the Expression of Heat Shock Protein 70

Although a variety of nanoparticles (NPs) functionalized with amphotericin B, an antifungal agent widely used in the clinic, have been studied in the last years their cytotoxicity profile remains elusive. Here we show that human endothelial cells take up high amounts of silica nanoparticles (SNPs) conjugated with amphotericin B (AmB) (SNP-AmB) (65.4 12.4 pg of Si per cell) through macropinocytosis while human fibroblasts internalize relatively low amounts (2.3 0.4 pg of Si per cell) because of their low capacity for macropinocytosis. We further show that concentrations of SNP-AmB and SNP up to 400 mg/mL do not substantially affect fibroblasts. In contrast, endothelial cells are sensitive to low concentrations of NPs (above 10 mg/mL), in particular to SNP-AmB. This is because of their capacity to internalize high concentration of NPs and high sensitivity of their membrane to the effects of AmB. Low-moderate concentrations of SNP-AmB (up to 100 mg/mL) induce the production of reactive oxygen species (ROS), LDH release, high expression of pro-inflammatory cytokines and chemokines (IL-8, IL-6, G-CSF, CCL4, IL-1b and CSF2) and high expression of heat shock proteins (HSPs) at gene and protein levels. High concentrations of SNP-AmB (above 100 ug/mL) disturb membrane integrity and kill rapidly human cells(60% after 5 h). This effect is higher in SNP-AmB than in SNP

Investigacion en enfermeria: Brasil y Portugal en la construccion de la identidad profesional

Objective: to investigate the construction of professional identity in Brazilian and Portuguese nursing research, according to the perception of their researchers, adopting Gaston Bachelard as a theoretical framework. Method: descriptive and exploratory research, with a qualitative approach, undertaken in Brazil and Portugal. Semistructured interviews were held with 17 nurse researchers from both countries, with experience in scientific research and doctorates and/or post-doctorates. Analysis of the data was undertaken in the following order: exploratory phase, selection of the units of meaning and process of codification of the units of analysis, with the respective construction of aprioristic categories. Results: two categories emerged for discussion: object and lines of research in nursing; and the political attitude of the researchers. Conclusion: the weak points in the construction of the professional identity need to be broken with, based on the undertaking of experimental investigations, with immediate responses for the health services and which meet the social requirements. Professional satisfaction is the best advertisement for the profession. Through professional qualification in nursing it will be possible to occupy better positions and undertake the role of leadership and the political attitude with greater apprpriateness.Objetivo: compreender a construção da identidade profissional na pesquisa em enfermagem de Brasil e de Portugal segundo a percepção de seus pesquisadores, adotando-se Gaston Bachelard como referencial teórico. Método: pesquisa descritiva, exploratória, de abordagem qualitativa, realizada no Brasil e Portugal. Adotou-se entrevista semiestruturada com 17 enfermeiros pesquisadores de ambos os países, com experiência em pesquisa científica e título de doutorado e/ou pós-doutorado. A análise dos dados obedeceu a seguinte ordem: fase exploratória, seleção das unidades de significado e processo de codificação das unidades de análise com a respectiva construção de categorias não-apriorísticas. Resultados: emergiram duas categorias para discussão: Objeto e linhas de pesquisa em enfermagem; e, Atitude política dos pesquisadores. Conclusão: as fragilidades na construção da identidade profissional precisam ser rompidas a partir do desenvolvimento de investigações experimentais, com respostas imediatas aos serviços de saúde e que atendam às demandas sociais. Profissional satisfeito é o maior marketing da profissão. Por meio da qualificação profissional na enfermagem será possível ocupar melhores cargos e desenvolver com maior propriedade o papel de liderança e a atitude política.percepción de sus investigadores, adoptándose Gaston Bachelard como referencial teórico. Método: investigación descriptiva, exploratoria, de abordaje cualitativo, realizado en Brasil y Portugal. Se adoptó entrevista semiestructurada con 17 enfermeros investigadores de ambos países, con experiencia en investigación científica y titulo de doctorado y/o post-doctorado. El análisis de los datos obedeció el siguiente orden: fase exploratoria, selección de las unidades de significado y proceso de codificación de las unidades de análisis con la respectiva construcción de categorías no apriorísticas. Resultados: emergieron dos categorías para discusión: Objeto y líneas de investigación en enfermería y; actitud política de los investigadores. Conclusión: las fragilidades en la construcción de la identidad profesional precisan ser rotas a partir del desarrollar de investigaciones experimentales, con respuestas inmediatas a los servicios de salud y que atiendan a las demandas sociales. Profesional satisfecho es el mayor marketing de la profesión Por medio de la calificación profesional en enfermería será posible ocupar mejores cargos y desarrollar con mayor propiedad el papel de liderazgo y la actitud política.info:eu-repo/semantics/publishedVersio

Influence of canopy fruit location on morphological, histochemical and biochemical changes in two oil olive cultivars

The influence of different irradiance conditions was evaluated under natural solar radiation by comparing well-exposed (in) and shaded fruit (out) in canopies of olive trees (Olea europaea L). Over a 2-year period, from 50 days after full bloom up to harvest time, “in” and “out” olive samples of two genotypes (“Frantoio Millennio” and “Coratina 5/19”) were periodically collected. Morphological, histochemical, and biochemical analysis were performed to study the changes on fruit morphometric traits, oil body accumulation, and b-glucosidase enzyme activity. Some parameters were modified by shading inside the canopy in which the proportion of incident photosynthetically active radiation intercepted by the crop was 47%. Shaded fruits developed at slow rate and were characterized by late darkgoing time, reduced size, with a tendency toward oblong shape. The rapid histochemical procedure proposed to estimate the oil body accumulation during fruit ripening showed that a reduced irradiance caused a decrease in oil body density. The canopy position influenced, in a different way, the b-glucosidase activity in relation to the fruit-ripening stage in both genotypes. These findings indicate that providing an adequate and uniform lighting of the olive canopy by careful choices of orchard management practices can be a key factor for several yield components

Endometrial apoptosis and neutrophil infiltration during menstruation exhibits spatial and temporal dynamics that are recapitulated in a mouse model.

Abstract Menstruation is characterised by synchronous shedding and restoration of tissue integrity. An in vivo model of menstruation is required to investigate mechanisms responsible for regulation of menstrual physiology and to investigate common pathologies such as heavy menstrual bleeding (HMB). We hypothesised that our mouse model of simulated menstruation would recapitulate the spatial and temporal changes in the inflammatory microenvironment of human menses. Three regulatory events were investigated: cell death (apoptosis), neutrophil influx and cytokine/chemokine expression. Well-characterised endometrial tissues from women were compared with uteri from a mouse model (tissue recovered 0, 4, 8, 24 and 48 h after removal of a progesterone-secreting pellet). Immunohistochemistry for cleaved caspase-3 (CC3) revealed significantly increased staining in human endometrium from late secretory and menstrual phases. In mice, CC3 was significantly increased at 8 and 24 h post-progesterone-withdrawal. Elastase+ human neutrophils were maximal during menstruation; Ly6G+ mouse neutrophils were maximal at 24 h. Human endometrial and mouse uterine cytokine/chemokine mRNA concentrations were significantly increased during menstrual phase and 24 h post-progesterone-withdrawal respectively. Data from dated human samples revealed time-dependent changes in endometrial apoptosis preceding neutrophil influx and cytokine/chemokine induction during active menstruation. These dynamic changes were recapitulated in the mouse model of menstruation, validating its use in menstrual research

Urokinase Plasminogen Receptor and the Fibrinolytic Complex Play a Role in Nerve Repair after Nerve Crush in Mice, and in Human Neuropathies

Remodeling of extracellular matrix (ECM) is a critical step in peripheral nerve regeneration. In fact, in human neuropathies, endoneurial ECM enriched in fibrin and vitronectin associates with poor regeneration and worse clinical prognosis. Accordingly in animal models, modification of the fibrinolytic complex activity has profound effects on nerve regeneration: high fibrinolytic activity and low levels of fibrin correlate with better nerve regeneration. The urokinase plasminogen receptor (uPAR) is a major component of the fibrinolytic complex, and binding to urokinase plasminogen activator (uPA) promotes fibrinolysis and cell movement. uPAR is expressed in peripheral nerves, however, little is known on its potential function on nerve development and regeneration. Thus, we investigated uPAR null mice and observed that uPAR is dispensable for nerve development, whereas, loss of uPAR affects nerve regeneration. uPAR null mice showed reduced nerve repair after sciatic nerve crush. This was a consequence of reduced fibrinolytic activity and increased deposition of endoneurial fibrin and vitronectin. Exogenous fibrinolysis in uPAR null mice rescued nerve repair after sciatic nerve crush. Finally, we measured the fibrinolytic activity in sural nerve biopsies from patients with peripheral neuropathies. We showed that neuropathies with defective regeneration had reduced fibrinolytic activity. On the contrary, neuropathies with signs of active regeneration displayed higher fibrinolytic activity. Overall, our results suggest that enforced fibrinolysis may facilitate regeneration and outcome of peripheral neuropathies