RESUMEN
Serum albumin contamination of cells cultured in vitro significantly impedes the mass spectrometric analysis of proteins secreted by the cells. Here we report a novel washing and culturing technique for rat vascular endothelial cells that considerably reduces the concentration of the commonly used additive for cell culture, bovine serum albumin (BSA), in the secretome of these cells. Cells are rinsed stringently and cultured for 24 h in serum-free media without appreciably impeding cell growth or viability. The percentage of BSA scans identified by tandem mass spectrometry (LC-MS/MS) in stringently rinsed cells (average 13.2%) was significantly lower than either the moderately rinsed or no rinse cell treatments (average 35.2% and 45.2% respectively). Furthermore, the stringent wash treatment allowed the confident identification of a larger portion of the secretome of rat endothelial cells by LC-MS/MS.
Asunto(s)
Técnicas de Cultivo de Célula , Células Endoteliales/química , Espectrometría de Masas , Proteoma/análisis , Proteómica/métodos , Animales , Bovinos , Células Cultivadas , Medios de Cultivo/química , Células Endoteliales/metabolismo , Endotelio Vascular/citología , Ratas , Albúmina Sérica Bovina/análisisRESUMEN
BACKGROUND: and Aims Nassella trichotoma is an unpalatable perennial grass weed that invades disturbed native grasslands in temperate regions of south-eastern Australia. This experiment investigated whether elevated N levels, often associated with disturbance, increases the competitiveness of N. trichotoma relative to C3 and C4 native Australian grasses. METHODS: A pot experiment investigated competitive interactions between four native grasses, two C3 species (Microlaena stipoides and Austrodanthonia racemosa) and two C4 species (Themeda australis and Bothriochloa macra), and N. trichotoma at three different N levels (equivalent to 0, 60 and 120 kg ha-1) and three competing densities (zero, one and eight neighbouring plants), using an additive design. KEY RESULTS: All native grasses were competitive with N. trichotoma at low N levels, but only M. stipoides was competitive at high N. High densities of native grasses (8:1) had a major competitive effect on N. trichotoma at all N levels. The competitive ranking of native grasses, across all N levels, on N. trichotoma was: M. stipoides>A. racemosa>B. macra>T. australis. The C3 species were generally more competitive than the C4 species and C4 grasses were not inherently more productive at low N levels, in contrast to the results of other studies. CONCLUSION: To resist invasion from N. trichotoma, these native grasses need to be maintained at a high density and/or biomass. The results do not support the theory that species such as N. trichotoma, with high tissues density, are always less competitive than those of low tissue density; in this case competitiveness depended on N levels. The ability of N. trichotoma to accumulate biomass at a higher rate than these native grasses, helps to explain why it is a major weed in disturbed Australian native grasslands.
Asunto(s)
Nitrógeno/metabolismo , Poaceae/metabolismo , Australia , Biomasa , Suelo , Especificidad de la EspecieRESUMEN
Endothelial cells (ECs) are subjected to oxidative stress during many pathological processes, including ischemia/reperfusion and general inflammation. In the present study, we examined the effects of oxidative stress on rates of apoptosis in EC cultures. We treated large and microvessel ECs with menadione for 1 h in vitro to simulate the most common physiological form of oxidative stress, exposure to O2*-. Capillary ECs were resistant to menadione-induced apoptosis when compared with large-vessel ECs. Treatment with 35 microM menadione resulted in an apoptotic rate of approximately 5% in capillary EC cultures compared with approximately 45% in large-vessel EC cultures. At higher concentrations of menadione (35-75 microM), both types of ECs exhibited a concentration-related increase in apoptosis. Necrotic cell death only became evident at menadione concentrations ranging from 75-100 microM for both cell types. The timing of the apoptotic response to a 1 h menadione exposure was very specific. For both EC types, peaks of apoptosis occurred in two distinct waves, at 6-8 and 18-22 h after treatment. Analysis of the events leading up to the first peak of apoptosis indicated that specific matrix metalloproteinases (MMPs) were activated, suggesting that MMPs may be involved in initiating the apoptotic process.
Asunto(s)
Apoptosis , Endotelio Vascular/metabolismo , Estrés Oxidativo , Superóxidos/metabolismo , Animales , Apoptosis/efectos de los fármacos , Bovinos , Células Cultivadas , Medios de Cultivo Condicionados/química , Relación Dosis-Respuesta a Droga , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/enzimología , Glutatión/metabolismo , Metaloproteinasas de la Matriz/metabolismo , Microscopía Fluorescente , Microscopía de Contraste de Fase , Estrés Oxidativo/efectos de los fármacos , Fenantrolinas/farmacología , Acetato Fenilmercúrico/análogos & derivados , Acetato Fenilmercúrico/farmacología , Factores de Tiempo , Vitamina K/farmacologíaRESUMEN
In the review of the literature, the author notes several discussions concerning collaboration between nursing education and service agencies for updating faculty clinical practice. This article addresses the Preceptor-Preceptee model for Faculty "Cross-training," including the permission process, method, needs analysis, and evaluation. A concluding statement by the author strongly recommends the utilization of the faculty practice model to enhance the faculty's competence in clinical nursing areas.
Asunto(s)
Modelos de Enfermería , Práctica del Docente de Enfermería , Preceptoría , Competencia Clínica , Bachillerato en Enfermería , Humanos , Medicina Interna/educación , Enfermería Perioperatoria/educaciónRESUMEN
Parathyroid carcinoma is an uncommon endocrine tumor. Its usual clinical presentation is that of primary hyperparathyroidism with elevated serum calcium and parathormone levels. Nonfunctioning carcinoma of the parathyroid gland with normal serum calcium levels is very rare. There are 12 reported cases in the literature. This paper is a case report of a nonfunctioning parathyroid carcinoma and a review of the literature.