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Biomedical subjects

P J Mitchell

Publications and source records attributed to P J Mitchell.

At least 55 records · Page 3Linked to original sources
PubMed1999

Nimodipine and perfusion changes after stroke.

BACKGROUND AND PURPOSE: Meta-analysis of previous trials of oral nimodipine in acute stroke has suggested a benefit when commenced within 12 hours of onset. We sought to study the effect of oral nimodipine on reperfusion after acute stroke and the relation between reperfusion and outcome. METHODS: Fifty patients with acute middle cerebral artery territory cortical infarction were blindly randomized within 12 hours of onset to either oral nimodipine (30 mg every 6 hours) or placebo. Treatment was continued for 2 weeks. Cerebral blood flow was assessed with the use of 99mTc-hexamethylpropyleneamine oxime single-photon emission CT before therapy, 24 hours later, and at 3 months. Hypoperfusion was measured by a validated volumetric technique. Neurological impairment and functional outcome were assessed with the Canadian Neurological Scale and Barthel Index, respectively. Tissue loss was measured with CT at 3 months. Four patients were excluded from analysis for technical reasons. RESULTS: Twenty-three patients received nimodipine, and 23 received placebo. In the nimodipine group, there was early reperfusion that was not maintained at outcome (P=0.01). In the placebo group, mean infarct hypoperfusion volumes showed no overall change. Nonnutritional reperfusion in nimodipine-treated patients was associated with adverse neurological (P=0.05) and functional outcome (P=0.06). There was, however, no difference in clinical outcome between the 2 groups. CONCLUSIONS: Oral nimodipine administered within 12 hours enhanced acute reperfusion, but this was largely nonnutritional. Larger studies using a shorter treatment delay are required to evaluate the clinical efficacy of nimodipine in acute ischemic stroke.

Acute Disease↗
PubMed1999

Regulation of Hoxa2 in cranial neural crest cells involves members of the AP-2 family.

Hoxa2 is expressed in cranial neural crest cells that migrate into the second branchial arch and is essential for proper patterning of neural-crest-derived structures in this region. We have used transgenic analysis to begin to address the regulatory mechanisms which underlie neural-crest-specific expression of Hoxa2. By performing a deletion analysis on an enhancer from the Hoxa2 gene that is capable of mediating expression in neural crest cells in a manner similar to the endogenous gene, we demonstrated that multiple cis-acting elements are required for neural-crest-specific activity. One of these elements consists of a sequence that binds to the three transcription factor AP-2 family members. Mutation or deletion of this site in the Hoxa2 enhancer abrogates reporter expression in cranial neural crest cells but not in the hindbrain. In both cell culture co-transfection assays and transgenic embryos AP-2 family members are able to trans-activate reporter expression, showing that this enhancer functions as an AP-2-responsive element in vivo. Reporter expression is not abolished in an AP-2(alpha) null mutant embryos, suggesting redundancy with other AP-2 family members for activation of the Hoxa2 enhancer. Other cis-elements identified in this study critical for neural-crest-specific expression include an element that influences levels of expression and a conserved sequence, which when multimerized directs expression in a broad subset of neural crest cells. These elements work together to co-ordinate and restrict neural crest expression to the second branchial arch and more posterior regions. Our findings have identified the cis-components that allow Hoxa2 to be regulated independently in rhombomeres and cranial neural crest cells.

Animals↗
PubMed1998

Recognition and management of catheter-induced pulmonary artery rupture.

BACKGROUND: Catheter-induced pulmonary artery rupture is a well-recognized complication of invasive monitoring, but the risk has not diminished. Although commonly associated with cardiopulmonary bypass, injuries also occur in intensive care. Definitive proof requires pulmonary angiography or autopsy. Many cases are never reported, and lesser injuries are probably under-diagnosed. METHODS: Seven cases fulfilling accepted diagnostic criteria discovered over 2 years are described in four groups illustrating the common modes of presentation: hemoptysis with hypoxemia, exsanguination, delayed recurrent hemorrhage, and bleeding with cardiopulmonary bypass. RESULTS: One patient had a planned elective operation deferred. Four patients were being monitored in intensive care. Two of them died of pulmonary artery rupture. Two other patients had bleeding on weaning from cardiopulmonary bypass. One settled with conservative treatment, the other survived after extracorporeal life support. Recognition and management are discussed, emphasizing means of avoiding pulmonary resection. CONCLUSIONS: Catheter-induced pulmonary artery rupture is unavoidable. Constant awareness is essential. A plan of management is presented. Extracorporeal life support may help to avoid pulmonary resection. Early pulmonary angiography is advocated for accurate diagnosis and to enable treatment by embolization.

Aged↗
PubMed1998

DAP-2, the Drosophila homolog of transcription factor AP-2.

Transcription factor AP-2 is essential for craniofacial, nervous system, and limb development in the mouse. We report here the cloning and expression pattern analysis of DAP-2, the Drosophila homolog of AP-2 family genes. DAP-2 is expressed in discrete regions of procephalic neuroectoderm, the brain, ventral nerve cord, and maxillary segment during Drosophila embryogenesis, and in the brain, optic lobes, ventral nerve cord, antenno-maxillary complex, and antennal and leg imaginal disks in third instar larvae. Protein sequence conservation and parallels between the embryonic expression patterns of DAP-2 and mammal ian AP-2 family genes indicate that transcription factor AP-2 has been structurally and functionally conserved during metazoan evolution.

Animals↗
PubMed1998

Detection of renal arteries with fast spin-echo magnetic resonance imaging.

With the increasing use of non-invasive imaging with MR and volumetric CT to evaluate renal arteries, the ability to accurately detect the number and state of native renal arteries becomes critical if conventional angiography is to be supplanted in these settings. The present study evaluated the utility of a fast spin-echo (FSE) T2-weighted sequence to detect the number and course of renal arteries and their ostia compared to conventional angiography. Ten patients underwent conventional catheter angiography either for renal artery stenosis evaluation or as potential renal donors. Each patient then had an MR study of the renal arteries and kidneys with FSE MR (TR = 4000 ms, TE = 102 ms, eight-echo train length, 5-mm-thick interleaved 128 phase encodes, superior and inferior saturation pulses, number of excitations (NEX) = 4, on a 1.5-T superconducting magnet (GE, Milwaukee, WI, USA). Images were reviewed by two 'blinded' radiologists and renal arteries were counted and their ostia were evaluated. Results were compared with angiography and inter- and intra-observer statistics were calculated. All 10 patients underwent MR successfully, nine for renal artery stenosis (RAS) evaluation and one was a renal donor. A total of 24 renal arteries were imaged in 19 kidneys. Fast spin-echo MR is 95% accurate (95% CI: 88-100%) in detection of renal arteries, with no statistical difference between FSE MR and catheter angiography (McNemar P = 0.0). Inter- and intra-observer statistics demonstrate good-to-excellent agreement in renal artery detection (kappa: 0.63-0.90). In one case of RAS evaluation an incidental adrenal mass was detected as the aetiology of the patient's hypertension. Fast spin-echo MR can be a useful adjunct as part of the imaging for renal arteries with MRI.

Female↗
PubMed1997

Characterisation and chromosome mapping of the human non receptor tyrosine kinase gene, brk.

The brk gene encodes a non-receptor protein tyrosine kinase that consists of single SH3, SH2 and catalytic domains. Although BRK shows strongest sequence similarity to members of the SRC family of PTKs, there are several key structural and regulatory differences that place it on its own amongst non-receptor PTKs. In this study we have isolated genomic DNA clones corresponding to the human brk locus and used these to determine the intron-exon structure of the brk gene. The genomic structure of brk consists of 8 exons, whose boundaries are distinct from other non-receptor PTK family members, again indicating a structural and functional divergence. Alternate splicing of the primary brk transcript generates a distinct mRNA which encodes a truncated protein consisting of an SH3 domain and a novel C-terminal proline rich sequence. Using an antiserum raised to the SH3 domain, we have demonstrated that the product of this alternate brk transcript is expressed in the human breast tumour cell line T-47D. We have previously reported that expression of a tumour derived brk cDNA in mouse embryonic fibroblasts and human mammary epithelial cells supports anchorage independent growth, and in the latter potentiates the mitogenic response to epidermal growth factor. The protein encoded by the genomic sequence derived from normal human tissue is identical to that encoded by the tumour derived cDNA, and therefore the altered growth regulation is not associated with mutations within brk. In addition, we have identified a 5' genomic region that has promoter activity. The brk gene has been assigned to chromosome 20q 13.3 [corrected] using fluorescence in situ hybridisation (FISH).

Amino Acid Sequence↗
PubMed1997

Loss of expression of receptor tyrosine kinase family genes PTK7 and SEK in metastatic melanoma.

Protein tyrosine kinases (PTKs) have been implicated in the development of many common human tumours including melanoma. Previously we isolated PTK gene sequences expressed in normal melanocytes. Here we examined expression of 9 of these genes in cell lines derived from defined stages of melanoma progression, by Northern blotting and in some cases immunoblotting. We also tested cells from 2 animal models of particular stages in progression, as well as uncultured biopsies of metastatic melanoma. The expression of 2 receptor kinase family members found in melanocytes, PTK7/CCK-4 and SEK/TYRO1, was decreased or lost in advanced melanomas. PTK7 mRNA was found in only 54% of melanoma cell lines and 20% of melanoma biopsies. Similarly, expression was lost in 2 advanced cell lines selected from an early melanoma line that did express PTK7 mRNA. SEK/TYRO1 expression was observed in 75% and 17% of cell lines from primary and metastastic melanomas, respectively. Conversely, mRNA for the non-receptor kinase PTK6/BRK was not detected in normal melanocytes or primary melanoma lines, but was found in 9% of metastatic melanoma cell lines.

Cell Adhesion Molecules↗
PubMed1997

Potentiation of the time-dependent, antidepressant-induced changes in the agonistic behaviour of resident rats by the 5-HT1A receptor antagonist, WAY-100635.

Acute and chronic antidepressant drug treatments respectively decrease and increase the aggressive behaviour of resident rats during encounters with unfamiliar conspecifics. We have now examined the effect of the 5-hydroxytryptamine1A receptor antagonist, WAY-100635, on fluoxetine-, paroxetine- or venlafaxine-induced changes in aggression. WAY-100635 (0.1 mg/kg), which did not modify behaviour when given alone, potentiated the venlafaxine (5.54 mg/kg)-induced reduction in aggression after acute treatment and, during chronic treatment, accelerated the fluoxetine (0.34 mg/kg/day)-induced increase in aggression, from day 5 to day 2. A similar change in time course was seen with paroxetine (0.33 mg/kg/day), although the increase in aggression was smaller. Venlafaxine (5.54 mg/kg/day, alone or co-administered with WAY-100635) increased aggression by day 2. During chronic treatment, therefore, venlafaxine, at the dose used, had a more rapid onset of action than either fluoxetine or paroxetine, whereas the fluoxetine- and paroxetine-, but not the venlafaxine-, induced increase in aggression was accelerated by WAY-100635. These studies further support the hypothesis that selective blockade of the 5-hydroxytryptamine1A receptor augments the effects of antidepressant drugs in an animal model predictive of antidepressant activity, presumably by concomitant blockade of the somatodendritic 5-hydroxytryptamine1A autoreceptor-mediated negative feedback system of serotonergic neurones.

Agonistic Behavior↗
PubMed1997

Conflicting bright light exposure during night shifts impedes circadian adaptation.

This simulated night shift field study compared high-intensity ("bright") light exposures designed to either facilitate or conflict with adaptation to a 9-h phase shift of the sleep/dark schedule. There were 7 days of baseline with night sleep followed by 8 night shifts with day sleep in a 2 x 2 design with factors bright light (facilitating vs. conflicting) and direction of shifted sleep/dark (delayed vs. advanced). A total of 32 subjects (8 in each group) were exposed to 3 h of bright light (about 5,000 lux) and 5 h of ordinary indoor room light of "dim" light (< 500 lux) during each 8-h night shift. The bright light was timed according to the light phase-response curve (PRC) to delay or advance rhythms; it was timed to occur either before or after the baseline body temperature minimum, which served as an estimate of the PRC crossover point between delays and advances. Core body temperature was measured continuously and demasked to determine daily temperature minima. Significantly more subjects showed large temperature rhythm phase shifts (> or = 6 h during the last 4 night shifts relative to baseline) with facilitating bright light compared to conflicting bright light as well as with delayed sleep/dark compared to advanced sleep/dark. The combination of facilitating bright light and delayed sleep/dark produced large phase delay shifts in all subjects tested. By contrast, the combination of conflicting bright light and advanced sleep/dark resulted in very small phase shifts in most subjects. Because bright light timed to delay usually was not able to phase shift rhythms when sleep/dark was advanced, it appears that the timing of sleep/dark was as important as the timing of the bright light. There was a relationship between the amount of phase shift and the individual's baseline phase when sleep/dark was delayed. Larger phase delays were achieved by subjects with later baseline temperature minima and greater eveningness on the Morningness-Eveningness Questionnaire. These results show that it is important to time bright light appropriately to achieve circadian adaptation to the night shift and that individual differences play an important role in the ability of the circadian system to phase shift.

Adaptation, Physiological↗
PubMed1996

Brk, a breast tumor-derived non-receptor protein-tyrosine kinase, sensitizes mammary epithelial cells to epidermal growth factor.

brk (breast tumor kinase) shows homology to the src family of non-receptor protein-tyrosine kinases and is expressed in breast carcinomas. In order to investigate the role of brk in breast tumor development, we have examined the growth and transformation properties of human mammary epithelial cells engineered to overexpress Brk. Interestingly, like c-Src, overexpression of Brk leads to sensitization to EGF, and also results in a partially transformed phenotype. Further investigation of the latter activity was attempted by mutational analysis, targeting key residues known to affect tyrosine kinase activity in Src-like kinases. Mutation of amino acid residue Lys-219 to Met, by analogy to Src, abolished both kinase activity and transformation capacity. Mutation of amino acid residue Tyr-447 to Phe, however, resulted in a decrease in transforming potential without affecting kinase activity. These results suggest that while Src and Brk share some functional properties, they act differently during transformation. These differences are discussed in the context of the mechanisms underlying breast cancer development.

3T3 Cells↗
PubMed1996

Transcription factor AP-2 essential for cranial closure and craniofacial development.

During closure of the neural tube in the mouse, transcription factor AP-2 is expressed in ectoderm and in neural-crest cells migrating from the cranial neural folds. Cranial neural crest cells provide patterning information for craniofacial morphogenesis, generate most of the skull bones, and together with placodal ectoderm, form the cranial ganglia. To study the role of AP-2 during embryogenesis, we undertook a targeted mutagenesis of the AP-2 gene in the mouse. Here we report that AP-2(-/-) mice died perinatally with cranio-abdominoschisis and severe dismorphogenesis of the face, skull, sensory organs and cranial ganglia. Failure of cranial closure between 9 and 9.5 days postcoitum coincided with increased apoptosis in the midbrain, anterior hindbrain and proximal mesenchyme of the first branchial arch, but did not involve loss of expression of twist or Pax-3, two other regulatory genes known to be required for cranial closure.

Animals↗
PubMed1996

Brain cell type specificity and gliosis-induced activation of the human cytomegalovirus immediate-early promoter in transgenic mice.

Human cytomegalovirus (HCMV) can cause debilitating, sometimes fatal, opportunistic infections in congenitally infected infants and in immunodeficient individuals such as patients with the acquired immunodeficiency syndrome (AIDS). Molecular mechanisms that determine cell type specificity of HCMV infection and latency are poorly understood. We recently described a transgenic mouse model for analysis of HCMV major immediate-early (IE) promoter regulation and showed that sites of IE promoter activity during murine embryogenesis correlate with known target tissues of congenital HCMV infection in human fetuses (Koedood et al., 1995). Among various permissive human tissues, the brain is a site where HCMV infections can be devastating. Here, we have used immunohistochemical double-labeling analysis to identify specific cell types with HCMV-IE promoter activity in brains of transgenic mice at several postnatal stages. IE promoter activity was restricted to some endothelial cells, ependymal cells, choroid plexus epithelia, and neurons at discrete locations in the forebrain, brainstem, and cerebellum. Endothelial cells and neurons with activity were proportionately more abundant in neonatal than in adult brains. Although the IE promoter was normally silent in most astrocytes, activity was strongly induced in reactive astrocytes in response to a neocortical stab lesion. The findings support a model, consistent with clinical literature on HCMV encephalitis, whereby tissue damage and gliosis caused by HCMV infection of endothelial and ependymal cells progressively renders adjacent permissive neurons and reactive astrocytes accessible to infection. This transgenic model system should facilitate identification of factors that regulate the HCMV IE promoter with regard to infection permissivity and reactivation from latency.

Animals↗
PubMed1996

Effects of the 5-HT1A antagonist (+)-WAY-100135 on murine social and agonistic behavior.

Compounds previously identified as 5-HT1A antagonists have subsequently been demonstrated to possess partial agonistic properties in models assessing somatodendritic autoreceptor function. This study examined the influences of (+)-WAY-100135, claimed to be the first selective 5-HT1A antagonist, on offensive behaviour in male mice. Employing a resident-intruder paradigm, administration of (+)-WAY-100135 (1.0-10.0 mg/kg s.c.) enhanced elements of resident offensive behaviour at 2.5 and 5.0 mg/kg but reduced such behaviour at 10.0 mg/kg. In comparison, resident defensive postures remained unchanged except for a significant increase in defensive sideways behaviour at 10.0 mg/kg. These effects were accompanied by reduced rearing behaviour across the dose range tested. Attend/approach behaviour was significantly reduced at the lowest, but increased at the highest, doses tested. Such results may reflect response competition rather than concomitant motor impairment. Given the dynamic behavioural interactions occurring in this paradigm, the increased offensive behaviour of the resident mice leads to enhanced defence and counter-attack by the intruder conspecifics. The results are discussed with reference to the current literature concerning the behavioural effects of other 5-HT1A antagonists.

Agonistic Behavior↗
PubMed1996

Streptokinase increases luxury perfusion after stroke.

BACKGROUND AND PURPOSE: Recent acute stroke trials have reported that intravenous streptokinase is associated with an increased risk of adverse outcomes. We aimed to study the effect of streptokinase on the nature of reperfusion and the relation between reperfusion and clinical outcome. METHODS: We studied 24 patients in the Australian Streptokinase Trial with acute middle cerebral cortical infarction using 99mTc-hexamethylpropyleneamine oxime single-photon emission CT. Eleven of the 24 patients were scanned before therapy and again 24 hours later. The remaining 13 were scanned once either before therapy (1 patient) or after therapy (12 patients). All patients had outcome scans after 3 months. Infarct hypoperfusion was measured with a validated volumetric technique. Neurological impairment and functional outcome were assessed with the Canadian Neurological Scale and the Barthel Index, respectively. RESULTS: Fifteen patients received streptokinase and 9 received placebo. There was no difference in early reperfusion between streptokinase and placebo. However, streptokinase was associated with a greater amount of nonnutritional reperfusion than was placebo (P = .04). This luxury perfusion was associated with poor functional outcome (P = .02). CONCLUSIONS: This study suggests that streptokinase augments luxury perfusion after stroke. Luxury perfusion is associated with a worse outcome, which might be due in part to reperfusion injury.

Adult↗
PubMed1996

Microemboli during carotid angiography. Association with stroke risk factors or subsequent magnetic resonance imaging changes?

BACKGROUND AND PURPOSE: Carotid angiography is associated with a 1% risk of major stroke. Recently, transcranial Doppler ultrasonography (TCD) has shown cerebral microemboli during carotid angiography. To determine their significance, we correlated the number of microemboli during angiography with clinical characteristics, angiography findings, and preangiography and postangiography cerebral magnetic resonance imaging (MRI). METHODS: One middle cerebral artery was monitored with TCD in 24 patients during angiography for carotid territory ischemia. The number of microemboli was correlated with angiographic and clinical characteristics. T2-weighted cerebral MRI was performed before and < or = 48 hours after angiography, and the number of new ischemic lesions was determined in a blinded review. RESULTS: Microemboli were seen in all patients, with an average of 51 per procedure (range, 12 to 154). The majority of microemboli had signal characteristics typical of air. Sixteen of the 24 patients had both preangiography and postangiography MRI. One of 24 patients had an angiographic stroke, with a single new thalamic lesion on MRI. No other patient had a new lesion. The microembolus count correlated with the angiographic contrast volume (P < .001) but not with any other radiological or clinical characteristic. CONCLUSIONS: This study confirmed the presence of numerous cerebral microemboli during carotid angiography. The microembolic signal characteristics and the correlation with contrast volume indicate that introduced air is the cause. These microemboli are usually clinically silent and do not lead to new changes on cerebral MRI.

Adult↗
PubMed1995

Pez: a novel human cDNA encoding protein tyrosine phosphatase- and ezrin-like domains.

We have isolated cDNAs from normal human breast tissue and breast tumour cells that encode a protein (pez) with features of a novel non-receptor tyrosine phosphatase possessing N-terminal sequence homology to the ezrin-band 4.1-merlin-radixin protein family. Northern blot analysis indicates that pez is expressed in a variety of human tissues including kidney, skeletal muscle, lung and placenta. Fluorescence in situ hybridization has mapped pez to chromosome 1 region q32.2-41. Sequence identity to a characterized polymorphic marker confirms this localization.

Amino Acid Sequence↗