Developing research skills in medical students: AMEE Guide No. 69.

This Guide has been written to provide guidance for individuals involved in curriculum design who wish to develop research skills and foster the attributes in medical undergraduates that help develop research. The Guide will provoke debate on an important subject, and although written specifically with undergraduate medical education in mind, we hope that it will be of interest to all those involved with other health professionals' education. Initially, the Guide describes why research skills and its related attributes are important to those pursuing a medical career.

Endophilin I expression is increased in the brains of Alzheimer disease patients.

Alzheimer patients have increased levels of both the 42 amyloid-beta-peptide (Abeta) and the amyloid binding alcohol dehydrogenase (ABAD), which is an intracellular binding site for Abeta. The overexpression of Abeta and ABAD in transgenic mice has shown that the binding of Abeta to ABAD results in amplified neuronal stress and impairment of learning and memory. From a proteomic analysis of the brains from these animals, we have identified for the first time that the protein endophilin I increases in Alzheimer diseased brain.

Interaction of amyloid binding alcohol dehydrogenase/Abeta mediates up-regulation of peroxiredoxin II in the brains of Alzheimer's disease patients and a transgenic Alzheimer's disease mouse model.

Alzheimer's patients have increased levels of both the 42 beta amyloid-beta-peptide (Abeta) and amyloid binding alcohol dehydrogenase (ABAD) which is an intracellular binding site for Abeta. The over-expression of Abeta and ABAD in transgenic mice has shown that the binding of Abeta to ABAD results in exaggerating neuronal stress and impairment of learning and memory. From a proteomic analysis of the brains from these animals we identified that peroxiredoxin II levels increase in Alzheimer's diseased brain.

The vomeronasal organ in the human embryo, studied by means of three-dimensional computer reconstruction.

The human vomeronasal organ is of interest because of its potential role in sex pheromone detection. Due to the scarcity of early human material, studies of its development have concentrated on fetal rather than embryonic stages. The availability of embryonic specimens in the Walmsley Collection has enabled us to study the development of the vomeronasal organ (VNO) in human embryos between Carnegie Stages 17 and 23.

Computer-aided interactive three-dimensional reconstruction of the embryonic human heart.

Despite the fact that development of the human embryo heart is of considerable clinical importance, there is still disagreement over the process and the timing of events. It is likely that some of the conflicting accounts may have arisen from difficulties in describing and visualising 3-dimensional structures from 2-dimensional sections. To help overcome this problem and to improve our understanding of the development of the heart, we have devised techniques for the production of interactive 3D models reconstructed from serial histological sections of human embryos.

Cartilaginous development of the human craniovertebral junction as visualised by a new three-dimensional computer reconstruction technique.

Serial transverse histological sections of the human craniovertebral junction (CVJ) of 4 normal human embryos (aged 45 to 58 d) and of a fetus (77 d) were used to create 3-dimensional computer models of the CVJ. The main components modelled included the chondrified basioccipital, atlas and axis, notochord, the vertebrobasilar complex and the spinal cord. Chondrification of the component parts of CVJ had already begun at 45 d (Stage 18).

The study of early human embryos using interactive 3-dimensional computer reconstructions.

Tracings of serial histological sections from 4 human embryos at different Carnegie stages were used to create 3-dimensional (3D) computer models of the developing heart. The models were constructed using commercially available software developed for graphic design and the production of computer generated virtual reality environments. They are available as interactive objects which can be downloaded via the World Wide Web.

World Wide Web access to the British Universities Human Embryo Database.

The British Universities Human Embryo Database has been created by merging information from the Walmsley Collection of Human Embryos at the School of Biological and Medical Sciences, University of St Andrews and from the Boyd Collection of Human Embryos at the Department of Anatomy, University of Cambridge. The database has been made available electronically on the Internet and World Wide Web browsers can be used to implement interactive access to the information stored in the British Universities Human Embryo Database. The database can, therefore, be accessed and searched from remote sites and specific embryos can be identified in terms of their location, age, developmental stage, plane of section, staining technique, and other parameters.

The World-Wide Web: an interface between research and teaching in bioinformatics.

The rapid expansion occurring in World-Wide Web activity is beginning to make the concepts of 'global hypermedia' and 'universal document readership realistic objectives of the new revolution in information technology. One consequence of this increase in usage is that educators and students are becoming more aware of the diversity of the knowledge base which can be accessed via the Internet. Although computerised databases and information services have long played a key role in bioinformatics these same resources can also be used to provide core materials for teaching and learning.

Incorporation of inositol into the phosphoinositides of lymphoblastoid cell lines established from bipolar manic-depressive patients.

Lymphoblastoid cell lines established from patients suffering from bipolar manic-depressive psychosis or from a control group have been used to study the metabolism of the polyphosphoinositides in these cells. Cells were incubated for up to 6 h in [3H]inositol and the extent of inositol incorporation into the mono-, di- and triphosphoinositides was measured after extracting the water- and lipid-soluble inositol-containing pools. Although both the uptake of inositol and the 'free' intracellular inositol pool sizes were similar in the two cell groups, the incorporation of [3H]inositol into the phosphoinositides of the cells derived from bipolar manic-depressives was significantly less (by around 50-60%) than that which occurred in the control cells.

Characterisation of atrial natriuretic peptide receptors in bovine ventricular sarcolemma.

Analysis of [125I]-ANP binding data in an isolated bovine ventricular sarcolemmal membrane fraction revealed a single high affinity binding site (Kd approximately 5 x 10(-11) M). The ring deleted ANP analogue des [QSGLG]-ANP (4-23)-NH2 bound with a 1000-fold lower affinity indicating the absence of C-type receptors in this preparation. ANP stimulated guanylate cyclase activity by up to 2-fold with half-maximal activation at approximately 10(-9) M.

Atrial natriuretic peptide receptors and activation of guanylate cyclase in rat cardiac sarcolemma.

Two classes of atrial natriuretic peptide (ANP) receptors are present in purified sarcolemmal membrane fractions isolated from rat ventricle. Scatchard analysis using [125I]-ANP reveals high affinity (Kd approximately 10(-11) M) and low affinity (Kd approximately 10(-9) M) binding sites. Basal guanylate cyclase activities associated with these membrane fractions range from 3.

Cation transport in lymphoblastoid cell lines established from bipolar manic-depressive patients.

Lymphoblastoid cell lines established from patients suffering from bipolar manic depression have been used to study the possible involvement of cation transport in the aetiology of this illness. No significant difference was found in the K+ fluxes mediated by the ouabain-sensitive sodium pump, the diuretic-sensitive cotransport system and the passive leak pathway of cell lines established from either control or bipolar subjects. The mean value for the specific binding of 3H-ouabain (sodium pump site number) was significantly higher in the bipolar group (approximately 30%) than in the control group.

Degradation of [125I]-atrial natriuretic peptide by a soluble metallopeptidase isolated from rat ventricular myocytes.

Atrial natriuretic peptide is rapidly degraded by a soluble, heat labile peptidase isolated from ventricular myocytes. Degradation of [125I]-ANP is antagonized by unlabelled ANP, bradykinin, glucagon, 1,10-phenanthroline, PCMB, EDTA and the bacterial antibiotic bacitracin, but not by phenylmethylsulphonyl fluoride, aprotinin, phosphoramidon, E-64, amastatin or the ACE inhibitor SQ 20881 and bradykinin potentiator C. In addition neither bovine serum albumin nor caesin afforded any protection against degradation.

Actions of atrial natriuretic peptide (ANP) on cyclic nucleotide concentrations and phosphatidylinositol turnover in ventricular myocytes.

Atrial natriuretic peptide (ANP) stimulates cGMP production in isolated rabbit ventricular myocytes incubated in the presence of the phosphodiesterase inhibitor isobutylmethylxanthine (1mM). Half maximal activation was found at 10(-8)M ANP. Cellular cGMP concentrations of around 0.

The effects of vanadate on rabbit ventricular muscle adenylate cyclase and sodium pump activities.

Vanadate in the +5 oxidation state has been reported to have a positive inotropic action on cardiac ventricular muscle. We have investigated the biochemical actions of vanadate on ventricular muscle adenylate cyclase and sodium pump activities in both intact or disrupted cell systems in an attempt to elucidate the mechanism(s) responsible for the physiological response. Vanadate at concentrations up to 100 microM (Ka = 2 microM) stimulated adenylate cyclase activity in sarcolemmal membrane preparations or disrupted myocytes isolated from rabbit ventricular muscle by 2-3-fold.

Role of passive potassium fluxes in cell volume regulation in cultured HeLa cells.

Cultured HeLa cells behave as ideal osmometers when subjected to hyperosmolar media, and show no volume regulatory behavior. In hypoosmolar solutions, cell swelling is not as great as predicted, and this is due largely to a loss of intracellular KCl. In hyperosmolar solutions there is a stimulation of the ouabain-insensitive but loop diuretic-sensitive 86Rb+ (K+) pathway.

An effect of piretanide upon the intracellular cation contents of cells subjected to partial chronic (Na-K) pump blockade by ouabain.

Cultured cells have been used to study the contribution made by the ouabain-insensitive but diuretic (piretanide)-sensitive K transport system (so-called cotransport) to the maintenance of intracellular Na+ and K+ contents in normal cells and in cells whose Na-pump sites have been subjected to chronic partial inhibition. In cells which have normally directed gradients of Na+ and K+, chronic incubation in piretanide (10(-4) M) for up to 24 hr has no significant effect on the internal ion contents of HeLa (human carcinoma), MDCK (dog kidney epithelium) or BC3H1 (mouse smooth muscle) cell lines. This observation is consistent with the notion that when the intracellular ion contents are in a normal steady state the net driving force acting upon the diuretic-sensitive K transport (Na + K + Cl cotransport system) is zero or very close to zero.

Biochemical and physiological adaptation to chronic propranolol treatment in the rat.

The biochemical and physiological aspects of isoprenaline sensitivity in normotensive rats were examined during and after abrupt withdrawal of chronic propranolol treatment. Serum propranolol concentrations in rats chronically treated for one month (0.125% propranolol in drinking water: 75-100 mg/kg/day) ranged from 7 to 23 ng/ml.

Effect of the serum concentration of the growth medium on the sodium pump site density of cultured HeLa cells.

The density of sodium pump sites in the plasma membrane of cultured HeLa cells has been measured as a function of the serum concentration of the cell growth medium. Growth in media containing increased concentrations of serum (from 1 to 20% v/v) leads to an increase in sodium pump site numbers (as measured by the specific binding of [3H]ouabain) and pump activity (as measured by the ouabain-sensitive 86Rb influx). Time-course studies show that (1) new sodium pump sites first appear some 3-6 h after transfer to medium containing an elevated serum concentration and (2) the serum-mediated increase in new sodium pump sites is completely abolished by the protein synthesis inhibitors, cycloheximide and actinomycin D.

Calcium elevation in cultured heart cells: its role in cell injury.

Inhibition of the Na+-K+ pump in cultured embryonic chick heart cells promotes the elevation of intracellular Ca2+ and is a useful manipulation to study the relationship between Ca2+ and myocardial cell injury. One hour of Na+-K+ pump inhibition resulted in a fourfold increase in cell Na+, a 50% decline in cell K+, and a 5- to 10-fold increase in cell Ca2+, 45% of which is mitochondrial. The degree of cell injury induced by Ca2+ loading was evaluated by monitoring the content of adenosine 5'-triphosphate (ATP) and the release of the intracellular enzyme lactate dehydrogenase (LDH).

Effect of ouabain upon diuretic-sensitive K+ transport in cultured cells. Evidence for separate modes of operation of the transporter.

(1) Unidirectional K+ (86Rb) influx and efflux were measured in subconfluent layers of MDCK renal epithelial cells and HeLa carcinoma cells. (2) In both MDCK and HeLa cells, the furosemide-inhibitable and chloride-dependent component of K+ influx/efflux was stimulated 2-fold by a 30 min incubation in 1 . 10(-3) M ouabain.