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A Easter

Publications and source records attributed to A Easter.

8 recordsLinked to original sources
PubMed2006

Optimisation and validation of a medium-throughput electrophysiology-based hERG assay using IonWorks HT.

INTRODUCTION: Regulatory and competitive pressure to reduce the QT interval prolongation risk of potential new drugs has led to focus on methods to test for inhibition of the human ether-a-go-go-related gene (hERG)-encoded K+ channel, the primary molecular target underlying this safety issue. Here we describe the validation of a method that combines medium-throughput with direct assessment of channel function. METHODS: The electrophysiological and pharmacological properties of hERG were compared using two methods: conventional, low-throughput electrophysiology and planar-array-based, medium-throughput electrophysiology (IonWorks HT). A pharmacological comparison was also made between IonWorks HT and an indirect assay (Rb+ efflux). RESULTS: Basic electrophysiological properties of hERG were similar whether recorded conventionally (HEK cells) or using IonWorks HT (CHO cells): for example, tail current V1/2 -12.1+/-5.0 mV (32) for conventional and -9.5+/-6.0 mV (46) for IonWorks HT (mean+/-S.D. (n)). A key finding was that as the number of cells per well was increased in IonWorks HT, the potency reported for a given compound decreased. Using the lowest possible cell concentration (250,000 cells/ml) and 89 compounds spanning a broad potency range, the pIC50 values from IonWorks HT (CHO-hERG) were found to correlate well with those obtained using conventional methodology (HEK-hERG)(r=0.90; p<0.001). Further validation using CHO-hERG cells with both methods confirmed the correlation (r=0.94; p<0.001). In contrast, a comparison of IonWorks HT and Rb+ efflux data with 649 compounds using CHO-hERG cells showed that the indirect assay consistently reported compounds as being, on average, 6-fold less potent, though the differences varied depending on chemical series. DISCUSSION: The main finding of this work is that providing a relatively low cell concentration is used in IonWorks HT, the potency information generated correlates well with that determined using conventional electrophysiology. The effect on potency of increasing cell concentration may relate to a reduced free concentration of test compound owing to partitioning into cell membranes. In summary, the IonWorks HT hERG assay can generate pIC50 values based on a direct assessment of channel function in a timeframe short enough to influence chemical design.

Animals↗
PubMed2001

Management of patients with multiple rib fractures.

Multiple rib fractures in trauma patients are associated with significant morbidity and mortality. Delayed morbidity for patients with rib fractures is often a result of hypoventilation leading to atelectasis, pneumonia, and respiratory failure. Pain management was first recognized as an important factor in preventing complications in these patients. Later, management of the respiratory system became more widely recognized as a major factor in patients' care. It is now known that patients with multiple rib fractures benefit most from adequate pain control, rapid mobilization, and meticulous respiratory care to prevent complications. A protocol based on a synthesis of the existing literature is developed. Development of such a protocol for decisions about rapid mobilization, respiratory support, and pain management is the first step in testing the hypothesis that these interventions will decrease the length of patients' stay in intensive care units.

Age Factors↗
PubMed2000

Modulation of calcium current by recombinant GABA(B) receptors.

Two GABA(B) receptor subunits have been cloned: GABA(B1) and GABA(B2). In this study we investigate the coupling of recombinant GABA(B) receptors to calcium channels in differentiated NG108-15 cells, which exhibit many similarities to neurones but in which functional GABA(B) receptors are normally absent. Transfection of GABA(B1) and GABA(B2) subunit cDNAs enables baclofen-mediated inhibition of different calcium channel subtypes and a component of this modulation is voltage-dependent. When transfected individually, GABA(B2), but not GABA(B1), is able to enhance calcium current inhibition over background levels. Further, an antisense oligodeoxynucleotide to GABA(B1) reduces the average functional response in cells transfected with GABA(B2) alone. Assuming that the functional receptor is heteromeric, this suggests that GABA(B1), but not GABA(B2), is expressed endogenously in NG108-15 cells.

Animals↗
PubMed2000

Construct analysis of four modes of being present.

The purpose of this construct analysis is to delineate the manner or modes of presence as used by nurses with their patients. The scope of the term presence encompasses four separate, distinct modes of presence. The term concept does not encompass the scope of the term. Therefore, the term construct is applied to demonstrate the differences in the four modes of being present. Model cases were extrapolated from personal experiences. Methods used in the construct development and construction include components of the Wilsonian method, an evolutionary view, and the hybrid model. Due to the breath and complexity of the interrelatedness of the four modes of being present, the author presents only a model case. To illustrate the attributes and consequences for the patient and the nurse in the interpersonal relationship for each of the four modes of being present, the author created models.

Empathy↗
PubMed1999

Mechanisms contributing to the deficits in hippocampal synaptic plasticity in mice lacking amyloid precursor protein.

Abnormal processing of amyloid precursor protein (APP), in particular the generation of beta-amyloid (Abeta) peptides, has been implicated in the pathogenesis of Alzheimer's disease. This study examined the consequences of deleting the APP gene on hippocampal synaptic plasticity, and upon the biophysical properties of morphologically identified neurones in APP-null mice. The hippocampus of APP-null mice had a characteristic increase in gliosis throughout the CA1 region and a disruption of staining for the dendritic marker MAP2 and the presynaptic marker synaptophysin. The disruption of MAP2 staining was associated with a significant reduction in overall dendritic length and projection depth of biocytin labeled CA1 neurones. In two groups of APP-null mice that were examined at 8-12 months, and 20-24 months of age, there was an impairment in the formation of long-term potentiation (LTP) in the CA1 region compared to isogenic age matched controls. This LTP deficit was not associated with an alteration in the amplitude of EPSPs at low stimulus frequencies (0.033 Hz) or facilitation during a 100 Hz stimulus train, but was associated with a reduction in post-tetanic potentiation. Paired-pulse depression of GABA-mediated inhibitory post-synaptic currents was also attenuated in APP-null mice. These data demonstrate that the impaired synaptic plasticity in APP deficient mice is associated with abnormal neuronal morphology and synaptic function within the hippocampus.

Aging↗
PubMed1997

Modulation of long-term potentiation in CA1 region of mouse hippocampal brain slices by GABAA receptor benzodiazepine site ligands.

Enhancement of GABAA receptor function with benzodiazepine (BZ) site agonists can disrupt memory formation and hippocampal synaptic plasticity. To investigate this further the effects of the agonist, flunitrazepam, were contrasted with that of the inverse agonist, methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM), on NMDA-dependent LTP induction in the CA1 region of mouse hippocampus. Under control conditions, a priming stimulus (10 stimuli at 100 Hz) potentiated e.p.s.p. slopes by 198%, and subsequent burst stimuli (4 x 10 events at 100 Hz every 20 sec) by 306%. This potentiation was blocked by the non-competitive NMDA receptor antagonist MK-801 and the glycine site antagonist L-701,324. Flunitrazepam (1 microM) alone caused a slight but significant reduction in e.p.s.p.s to 83% of control, suppressed LTP induced by priming stimuli (133%) and burst stimuli (188%), but not that induced by sustained high-frequency stimulation (2 x 100 events at 100 Hz, 20 sec apart). The suppression of LTP induction by flunitrazepam was blocked by the benzodiazepine site antagonist flumazenil. In contrast, the inverse agonist DMCM (100 nM) potentiated LTP formed by both priming (to 283%) and burst stimuli (to 477%). This was associated with an enhancement of paired pulse facilitation during the induction phase and the subsequent appearance of paroxysmal burst discharges. Therefore, in addition to improvements in learning and memory as a result of improved vigilance, benzodiazepine inverse agonists can have direct effects on synaptic processes thought to contribute to memory formation.

Animals↗
PubMed1997

The state of research on the effects of therapeutic touch.

Therapeutic Touch is investigated using an integrative review of the literature. Using Ganong's (1987) methodology, the article explores the research question, What is the state of development of research regarding Therapeutic Touch? by analyzing primary research reports from 23 articles in 14 referred journals. The findings of the review indicate positive regard for the use of Therapeutic Touch. All research points to the need for further study in this area. Research methods used are satisfactory, but more rigorous methodologies would promote a more scientific contribution to the body of literature on Therapeutic Touch.

Data Interpretation, Statistical↗