The formative impact of general practice appraisals: a post-appraisal questionnaire survey.

Since their national implementation in 2002, GP appraisals are being extensively applied. There is a positive link between appraisals and patient care in hospitals. There also appears to be no uniform model for appraisals, which has led to confusion between their 'stand-alone' role as a formative assessment tool and their contribution to summative revalidation. This is a case study into the formative element of appraisals as carried out at a primary care trust in Wessex. We offer an insight into the diversity of learning outcomes and developmental potential of the formative element of appraisals (excluding revalidation) attained through the analysis of emerging themes from responses to a purposely designed questionnaire aimed at appraisees (questions appear in the tables). To our knowledge this is one of the few research articles that deal with the formative educational element of GP appraisals.

The Foundation Programme in general practice: the value added of the attachment - a Wessex experience.

The general practice (GP) educational arena offers a range of learning opportunities for junior doctors in the Foundation Programme (FP). In this study we explored what might be the value added of the experience in GP for current FP trainees in their second year of training (F2s) in Wessex. This is particularly important in view of the changes introduced by the new form of training which include a competence-based curriculum, new assessment forms, and shorter length of training than previously experienced by GP trainers in Wessex (four months). The aim of our study was therefore to explore what might be the contribution of the GP attachment to the formation of FP doctors in Wessex. We collected data from trainers and trainees on their perspectives of the GP attachment using two distinct qualitative questionnaires. We show that trainees in Wessex benefit from the experience in a variety of ways. Variations on the learning opportunities offered in GP may depend on the location of the practice, social environment, cases reviewed, teaching styles, perceptions on the new curriculum and assessment, and the general management of the practice.

Cell shape-dependent Control of Ca2+ influx and cell cycle progression in Swiss 3T3 fibroblasts.

The ability of adherent cells such as fibroblasts to enter the cell cycle and progress to S phase is strictly dependent on the extent to which individual cells can attach to and spread on a substratum. Here we have used microengineered adhesive islands of 22 and 45 mum diameter surrounded by a nonadhesive substratum of polyhydroxyl methacrylate to accurately control the extent to which individual Swiss 3T3 fibroblasts may spread. The effect of cell shape on mitogen-evoked Ca2+ signaling events that accompany entry into the cell cycle was investigated. In unrestricted cells, the mitogens bombesin and fetal calf serum evoked a typical biphasic change in the cytoplasmic free Ca2+ concentration. However, when the spreading of individual cells was restricted, such that progression to S phase was substantially reduced, both bombesin and fetal calf serum caused a rapid transient rise in the cytoplasmic free Ca2+ concentration but failed to elicit the normal sustained influx of Ca2+ that follows Ca2+ release. As expected, restricting cell spreading led to the loss of actin stress fibers and the formation of a ring of cortical actin. Restricting cell shape did not appear to influence mitogen-receptor interactions, nor did it influence the presence of focal adhesions. Because Ca2+ signaling is an essential component of mitogen responses, these findings implicate Ca2+ influx as a necessary component of cell shape-dependent control of the cell cycle.

A nonradioactive assay for nitric oxide synthase activity in tissue extracts.

We describe a fluorescence assay for nitric oxide synthase activity based on a new indicator, 4,5-diaminofluorescein (DAF-2). The method offers the advantage of being safer and more convenient than the citrulline radioassay in common use. The rapid and irreversible binding of DAF-2 to oxidized nitric oxide (NO) enables NO production to be measured in real time. The protocol is applied to the measurement of nitric oxide synthase in crude extracts of skeletal muscle.

Atrial natriuretic peptide attenuates Ca2+ oscillations and modulates plasma membrane Ca2+ fluxes in rat hepatocytes.

BACKGROUND & AIMS: Oscillations in cytosolic free Ca2+ concentration are a fundamental mechanism of intracellular signaling in hepatocytes. The aim of this study was to examine the effects of atrial natriuretic peptide (ANP) on cytosolic Ca2+ oscillations in rat hepatocytes. METHODS: Cyclic guanosine monophosphate (cGMP) was measured by enzyme immunoassay. Cytosolic Ca2+ oscillations were recorded from single aequorin-injected hepatocytes. Ca2+ efflux from hepatocyte populations was measured by using extracellular fura-2. Ca2+ influx was estimated by Mn2+ quench of fluorescence of fura-2 dextran injected into single hepatocytes. RESULTS: ANP attenuated cytosolic Ca2+ oscillations through a decrease in their frequency. In addition, ANP dramatically stimulated plasma membrane Ca2+ efflux and modestly inhibited basal Ca2+ influx. All of the observed effects of ANP were mimicked by the cGMP analogue 8-bromo-cGMP (8-Br-cGMP), and were prevented by inhibition of protein kinase G. In contrast, activation of cytosolic guanylyl cyclase by sodium nitroprusside had no effect on Ca2+ efflux, Ca2+ influx, or Ca2+ oscillations. CONCLUSIONS: ANP decreases the frequency of Ca2+ oscillations and modulates plasma membrane Ca2+ fluxes in rat hepatocytes. Attenuation of oscillatory Ca2+ signaling in hepatocytes may represent a key role for ANP in vivo.