Biomedical subjects
M Parsons
Publications and source records attributed to M Parsons.
Darifenacin in the treatment of overactive bladder.
Antimuscarinic drug therapy has been shown to be effective in the management of patients with symptoms of the overactive bladder syndrome (OAB), but the bothersome antimuscarinic adverse effects of dry mouth, constipation, somnolence and blurred vision often affect compliance with medication. The development of bladder selective M3 specific antagonists offers the possibility of increasing efficacy whilst minimising adverse effects. The M3 specific antagonist solifenacin has recently been marketed, and darifenacin will soon be available. The purpose of this article is to review the pharmacology and clinical trial data available for darifenacin, in addition to examining its role in the treatment of the OBS.
Refining the perfusion-diffusion mismatch hypothesis.
BACKGROUND AND PURPOSE: The Echoplanar Imaging Thrombolysis Evaluation Trial (EPITHET) tests the hypothesis that perfusion-weighted imaging (PWI)-diffusion-weighted imaging (DWI) mismatch predicts the response to thrombolysis. There is no accepted standardized definition of PWI-DWI mismatch. We compared common mismatch definitions in the initial 40 EPITHET patients. METHODS: Raw perfusion images were used to generate maps of time to peak (TTP), mean transit time (MTT), time to peak of the impulse response (Tmax) and first moment transit time (FMT). DWI, apparent diffusion coefficient (ADC), and PWI volumes were measured with planimetric and thresholding techniques. Correlations between mismatch volume (PWIvol-DWIvol) and DWI expansion (T2(Day 90-vol)-DWI(Acute-vol)) were also assessed. RESULTS: Mean age was 68+/-11, time to MRI 4.5+/-0.7 hours, and median National Institutes of Health Stroke Scale (NIHSS) score 11 (range 4 to 23). Tmax and MTT hypoperfusion volumes were significantly lower than those calculated with TTP and FMT maps (P<0.001). Mismatch > or =20% was observed in 89% (Tmax) to 92% (TTP/FMT/MTT) of patients. Application of a +4s (relative to the contralateral hemisphere) PWI threshold reduced the frequency of positive mismatch volumes (TTP 73%/FMT 68%/Tmax 54%/MTT 43%). Mismatch was not significantly different when assessed with ADC maps. Mismatch volume, calculated with all parameters and thresholds, was not significantly correlated with DWI expansion. In contrast, reperfusion was correlated inversely with infarct growth (R=-0.51; P=0.009). CONCLUSIONS: Deconvolution and application of PWI thresholds provide more conservative estimates of tissue at risk and decrease the frequency of mismatch accordingly. The precise definition may not be critical; however, because reperfusion alters tissue fate irrespective of mismatch.
Imaging protein-protein interactions in cell motility using fluorescence resonance energy transfer (FRET).
Protein-protein interactions and signal transduction pathways have traditionally been analysed using biochemical techniques or standard microscopy. Although invaluable in the delineation of protein hierarchy, these methods do not provide information on the true spatial and temporal nature of complex formation within the intact cell. Recent advances in microscopy have allowed the development of new methods to analyse protein-protein interactions at very high resolution in both fixed and live cells. The present paper provides a brief overview of using fluorescence resonance energy transfer to analyse directly molecular interactions and conformational changes in various proteins involved in the regulation of cell adhesion and motility.
Safety and efficacy of chimeric yellow Fever-dengue virus tetravalent vaccine formulations in nonhuman primates.
To construct chimeric YF/DEN viruses (ChimeriVax-DEN), the premembrane (prM) and envelope (E) genes of yellow fever (YF) 17D virus were replaced with those of each wild-type (WT) dengue (DEN) virus representing serotypes 1 to 4. ChimeriVax-DEN1-4 vaccine viruses were prepared by electroporation of Vero cells with RNA transcripts prepared from viral cDNA (F. Guirakhoo, J. Arroyo, K. V. Pugachev, C. Miller, Z.-X. Zhang, R. Weltzin, K. Georgakopoulos, J. Catalan, S. Ocran, K. Soike, M. Ratteree, and T. P. Monath, J. Virol. 75:7290-7304, 2001; F. Guirakhoo, K. Pugachev, J. Arroyo, C. Miller, Z.-X. Zhang, R. Weltzin, K. Georgakopoulos, J. Catalan, S. Ocran, K. Draper, and T. P. Monath, Virology 298:146-159, 2002). Progeny viruses were subjected to three rounds of plaque purifications to produce the Pre-Master Seed viruses at passage 7 (P7). Three further passages were carried out using U.S. current Good Manufacturing Practices (cGMP) to produce the Vaccine Lot (P10) viruses. Preclinical studies demonstrated that the vaccine candidates are replication competent and genetically stable and do not become more neurovirulent upon 20 passages in Vero cells. The safety of a tetravalent vaccine was determined and compared to that of YF-VAX in a formal monkey neurovirulence test. Brain lesions produced by the tetravalent ChimeriVax-DEN vaccine were significantly less severe than those observed with YF-VAX. The immunogenicity and protective efficacy of four different tetravalent formulations were evaluated in cynomolgus monkeys following a single-dose subcutaneous vaccination followed by a virulent virus challenge 6 months later. All monkeys developed low levels of viremia postimmunization, and all the monkeys that had received equal concentrations of either a high-dose (5,5,5,5) or a low-dose (3,3,3,3) formulation seroconverted against all four DEN virus serotypes. Twenty-two (92%) of 24 monkeys were protected as determined by lack of viremia post-challenge. This report is the first to demonstrate the safety of a recombinant DEN virus tetravalent vaccine in a formal neurovirulence test, as well as its protective efficacy in a monkey challenge model.
Perfusion thresholds in acute stroke thrombolysis.
BACKGROUND AND PURPOSE: Perfusion-weighted MRI has been shown to be useful in the early identification of cerebral tissue at risk of infarction during acute ischemia. Identification of threshold perfusion measures that predict infarction may assist in the selection of patients for thrombolysis. METHODS: Mean transit time (MTT), regional cerebral blood flow (rCBF), and regional cerebral blood volume (rCBV) maps were generated in 35 acute stroke patients (17 treated with tissue plasminogen activator and 18 control patients) imaged within 6 hours from symptom onset. Day 90 outcome infarcts (T2-weighted MRI) were superimposed on acute MTT, rCBF, and rCBV maps. Perfusion-weighted MRI measures were then calculated for 2 regions: infarcted and salvaged tissue. RESULTS: MTT was prolonged by 22% in infarcted regions relative to salvaged tissue (P<0.001). rCBF was 10% lower in infarcted tissue than in salvaged regions (P<0.01). rCBV did not differ significantly between infarcted and salvaged regions. When reperfusion occurred, tissue with more severely prolonged MTT was salvaged from infarction relative to patients with persistent hypoperfusion (P<0.05). In contrast, rCBF in salvaged regions did not differ between patients with and without reperfusion. In reperfused patients, an inverse correlation (R=0.93, P<0.001) was found between time of initial MRI scan and MTT delay in salvaged tissue. CONCLUSIONS: Both increases in MTT and decreases in rCBF predict infarction. Differences in MTT also predict salvage in more severely hypoperfused tissue after reperfusion, suggesting that it is the most clinically useful quantitative perfusion measure. Perfusion thresholds for infarction need to be assessed in the context of symptom duration.
Is Doppler ultrasound sufficient as the sole investigation before carotid endarterectomy?
BACKGROUND AND AIMS: Doppler ultrasound (DU) of the extracranial carotid arteries has been advocated as the sole imaging modality in carotid endarterectomy (CE) candidates. However this approach fails to identify patients with potentially significant intracranial disease, at high risk of stroke and death. Therefore, many stroke clinicians recommend angiography after screening DU. We aimed to identify the proportion of cases referred for CE in whom the identification of intracranial disease could have altered management. METHODS: Two neuroradiologists, blinded to the clinical history, reviewed the films of 111 CE candidates, predominantly of Caucasian background, who had undergone carotid angiography after screening DU. Intracranial stenoses >50% luminal diameter, incidental aneurysms and non-atherosclerotic lesions were documented. Demographic and epidemiological data were collected. RESULTS: Of the 111 patients, 87 had >50% extracranial stenoses although two thirds were asymptomatic. Intracranial stenotic lesions were recorded in 29% of patients. Over half of these were tandem lesions, distal to an extracranial stenosis. Aneurysms were found in 4.5% of patients. CONCLUSIONS: DU alone would have failed to detect significant intracranial disease in nearly a third of cases. These patients are at high risk of stroke. The identification of this group allows more aggressive stroke prevention therapy.
Integrin-protein kinase C relationships.
The integrins have an ability to interact with extracellular matrix proteins to confer adhesive and motile properties on cells. The means by which these activities operate and the manner in which they are integrated with cell functions is of particular relevance to many biological processes. In the present paper, the developing understanding of the bi-directional relationship between the protein kinase C family of signal transducers and integrins is discussed.
The enhanced spontaneous activity of the diabetic colon is not the consequence of impaired inhibitory control mechanisms.
1. Approximately 45% of patients with diabetes mellitus have gastrointestinal complications such as diarrhoea and constipation, but the underlying aetiology is unclear. The present study investigates alterations in spontaneous motility of the colon that may be, in part, responsible for these symptoms using an established animal model of diabetes. 2. Rats were rendered diabetic by a single intraperitoneal injection of streptozotocin and age-matched controls were injected with citrate buffer. Rats were sacrificed after 8-weeks and proximal colonic circular muscle tissue mounted in organ baths. 3. Spontaneous activity was observed in both control and diabetic tissues, but this activity was almost doubled in colonic tissue taken from diabetic rats. It was hypothesized that this increase was due to a deficit in inhibitory control of the colon in the diabetic state. 4. Possible alterations in nitrergic and vasoactive intestinal polypeptide (VIP)ergic control were investigated using a range of pharmacological tools. 5. Sodium nitroprusside, VIP and antioxidants (reduced glutathione, ascorbate and alpha-tocopherol) inhibited the spontaneous activity, but the level of inhibition observed was not different in diabetic tissue compared with control. 6. Arginine, [D-p-Cl-Phe6, Leu17]-VIP and alpha-chymotrypsin had no effect on spontaneous activity in either sets of tissue. 7. N omega-nitro-L-arginine produced a small, but significant, increase in the level of spontaneous activity, but the degree of increase was not different between control and diabetic tissues. 8. Western blots demonstrated that there was no inducible-nitric oxide synthase (iNOS) in control or diabetic tissues and that the levels of endothelial-NOS (eNOS) and neuronal-NOS (nNOS) detected were not statistically significantly different. The [3H]-citrulline assay established that the functionality of the NOS isoforms present were unaltered in the diabetic state. 9. This study demonstrates that there is a marked alteration in motility in the colon taken from diabetic animals. However, the change in motility is unlikely to be due to a change in inhibitory control mechanisms and may be due to an increased excitability.
Changes in expression of P2 receptors in rat and mouse pancreas during development and ageing.
In view of the evidence for a role for extracellular ATP in both pancreatic endocrine and exocrine functions, we have investigated the expression of P2X and P2Y receptors in this tissue in neonate and aged rat and mouse. Using immunohistochemistry it was shown that P2X(1), P2X(4), P2X(7), P2Y(1) and P2Y(2) receptors were present in different regions of the rat and mouse pancreas; P2X(3) and P2X(6) receptors were not found, and P2X(5) immunolabelling was only found in some nerves. The pancreatic vasculature of both rat and mouse expressed P2X(1), P2X(2), P2Y(1) and P2Y(2) receptors in the smooth muscle. P2X(1) and P2X(4) receptors were absent in the islets of the neonate pancreas, but were progressively upregulated with age after birth. In contrast, the greatest expression of P2Y(1) in cells from the duct system was in neonate pancreas, while there was no P2Y(1) expression in aged rat pancreas. P2X(7) receptors had a consistent pattern of distribution in all of the groups examined, being located in the outer periphery of the islet. Using antibodies raised against insulin, somatostatin and glucagon, double-labelling immunofluorescence was used to identify P2X(7)-positive cells in different islet of Langerhans cell populations. Our results demonstrated a clear immunoreaction to P2X(7) receptors in islet alpha cells, while no P2X(7) was expressed in beta and delta cells. The significance of the differential expression of P2 receptors in the pancreas during development and ageing, and a possible role for the proliferation and death of the islet cell population are discussed.
A three-item scale for the early prediction of stroke recovery.
BACKGROUND: Accurate assessment of prognosis in the first hours of stroke is desirable for best patient management. We aimed to assess whether the extent of ischaemic brain injury on magnetic reasonance diffusion-weighted imaging (MR DWI) could provide additional prognostic information to clinical factors. METHODS: In a three-phase study we studied 66 patients from a North American teaching hospital who had: MR DWI within 36 h of stroke onset; the National Institutes of Health Stroke Scale (NIHSS) score measured at the time of scanning; and the Barthel Index measured no later than 3 months after stroke. We used logistic regression to derive a predictive model for good recovery. This logistic regression model was applied to an independent series of 63 patients from an Australian teaching hospital, and we then developed a three-item scale for the early prediction of stroke recovery. FINDINGS: Combined measurements of the NIHSS score (p=0.01), time in hours from stroke onset to MR DWI (p=0.02), and the volume of ischaemic brain tissue on MR DWI (p=0.04) gave the best prediction of stroke recovery. The model was externally validated on the Australian sample with 0.77 sensitivity and 0.88 specificity. Three likelihood levels for stroke recovery-low (0-2), medium (3-4), and high (5-7)-were identified on the three-item scale. INTERPRETATION: The combination of clinical and MR DWI factors provided better prediction of stroke recovery than any factor alone, shortly after admission to hospital. This information was incorporated into a three-item scale for clinical use.
Ezrin is a downstream effector of trafficking PKC-integrin complexes involved in the control of cell motility.
Protein kinase C (PKC) alpha has been implicated in beta1 integrin-mediated cell migration. Stable expression of PKCalpha is shown here to enhance wound closure. This PKC-driven migratory response directly correlates with increased C-terminal threonine phosphorylation of ezrin/moesin/radixin (ERM) at the wound edge. Both the wound migratory response and ERM phosphorylation are dependent upon the catalytic function of PKC and are susceptible to inhibition by phosphatidylinositol 3-kinase blockade. Upon phorbol 12,13-dibutyrate stimulation, green fluorescent protein-PKCalpha and beta1 integrins co-sediment with ERM proteins in low-density sucrose gradient fractions that are enriched in transferrin receptors. Using fluorescence lifetime imaging microscopy, PKCalpha is shown to form a molecular complex with ezrin, and the PKC-co-precipitated endogenous ERM is hyperphosphorylated at the C-terminal threonine residue, i.e. activated. Electron microscopy showed an enrichment of both proteins in plasma membrane protrusions. Finally, overexpression of the C-terminal threonine phosphorylation site mutant of ezrin has a dominant inhibitory effect on PKCalpha-induced cell migration. We provide the first evidence that PKCalpha or a PKCalpha-associated serine/threonine kinase can phosphorylate the ERM C-terminal threonine residue within a kinase-ezrin molecular complex in vivo.
A healthy old age: realistic or futile goal? Exercise programmes benefit even those who are most severely disabled.
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Biogenesis and function of peroxisomes and glycosomes.
Peroxisomes of higher eukaryotes, glycosomes of kinetoplastids, and glyoxysomes of plants are related microbody organelles that perform differing metabolic functions tailored to their cellular environments. The close evolutionary relationship of these organelles is most clearly evidenced by the conservation of proteins involved in matrix protein import and biogenesis. The glycosome can be viewed as an offshoot of the peroxisomal lineage with additional metabolic functions, specifically glycolysis and purine salvage. Within the parasitic protozoa, only kinetoplastids have been conclusively demonstrated to possess glycosomes or indeed any peroxisome-like organelle. The importance of glycosomal pathways and their compartmentation emphasizes the potential of the glycosome and glycosomal proteins as drug targets.
Policy or tradition: oral intake in labour.
Oral intake restrictions have varied over time and around the world with written hospital policies for this issue often being non-existent. As there are differing views on this issue within hospitals overseas, a survey was conducted of 109 maternity units in New South Wales, Australia during early 2000 to identify the trends across the state. In New South Wales 81.7% of hospitals did not have a written policy for oral intake in labour. The remaining 18.3% had written policies which varied in their oral intake allowances from ice only to whatever women feel like eating and drinking. Of the 109 hospitals in this survey 60.5% leave food and fluid requirements to the individual woman's discretion, providing they have no increased risk of general anaesthetic.
Evaluation of critical thinking outcomes of a BSN program.
Following a curriculum revision, which emphasized critical thinking, a school of nursing selected the California Critical Thinking Skills Test (CCTST) as a standardized outcomes measure for its bachelor's of science in nursing (BSN) program. Students in three tracks of the program were administered the CCTST on entry into the nursing curriculum and again on exit. Paired t tests for dependent samples were used to analyze pretest-posttest differences of all students (N = 136) in the program. Results for students in each of the three tracks demonstrated significantly improved (P < or = .05) CCTST scores on all subscales and total scores, with one exception. RN to BSN students' scores on the Analysis subscale approached but did not reach significance (P = .058). Implications for evaluation are discussed.
A novel nucleolar G-protein conserved in eukaryotes.
We describe here a novel, evolutionarily conserved set of predicted G-proteins. The founding member of this family, TbNOG1, was identified in a two-hybrid screen as a protein that interacts with NOPP44/46, a nucleolar phosphoprotein of Trypanosoma brucei. The biological relevance of the interaction was verified by co-localization and co-immunoprecipitation. TbNOG1 localized to the trypanosome nucleolus and interacted with domains of NOPP44/46 that are found in several other nucleolar proteins. Genes encoding proteins highly related to TbNOG1 are present in yeast and metazoa, and related G domains are found in bacteria. We show that NOG1 proteins in humans and Saccharomyces cerevisae are also nucleolar. The S. cerevisae NOG1 gene is essential for cell viability, and mutations in the predicted G motifs abrogate function. Together these data suggest that NOG1 may play an important role in nucleolar functions. The GTP-binding region of TbNOG1 is similar to those of Obg and DRG proteins, which, together with NOG, form a newly recognized family of G-proteins, herein named ODN. The ODN family differs significantly from other G-protein families, and shows several diagnostic sequence characteristics. All organisms appear to possess an ODN gene, pointing to the biological significance of this family of G-proteins.