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P J Morgan

Publications and source records attributed to P J Morgan.

At least 19 recordsLinked to original sources
PubMed2003

Precursor-protein convertase 1 gene expression in the mouse hypothalamus: differential regulation by ob gene mutation, energy deficit and administration of leptin, and coexpression with prepro-orexin.

The expression of precursor-protein convertase (PC)1, PC2 and paired basic amino acid cleaving enzyme four mRNA was studied by in situ hybridisation in regions of the hypothalamus involved in energy regulation in relation to obese (ob) gene mutation and energy deficit. PC1 gene was differentially expressed in hypothalamic nuclei of mice from different genetic backgrounds or energetic status, whereas no differences in expression were observed for either the PC2 or paired basic amino acid cleaving enzyme four genes. In obese ob/ob mice, PC1 mRNA levels were increased in the paraventricular nucleus, decreased in the lateral hypothalamus and unchanged in the ventromedial nucleus and arcuate nucleus relative to lean controls. In response to intraperitoneal injection of murine leptin, PC1 mRNA levels in obese ob/ob mice decreased in the arcuate nucleus, increased in the lateral hypothalamus and were unchanged in both the paraventricular nucleus and ventromedial nucleus. In mice deprived of food for 24 h, PC1 mRNA levels were reduced in the ventromedial nucleus, increased in the lateral hypothalamus and unchanged in the paraventricular nucleus and arcuate nucleus relative to ad libitum-fed controls. Overall, whilst the data show effects related to leptin and energetic status, they do not support a strong and consistent link between PC1 gene expression and energy balance. This suggests that if PC1 is important to the control of energy balance then protein expression and activity, rather than gene expression may be the more critical parameters of regulation. The relationship between PC1 and candidate energy balance-related genes in the lateral hypothalamus was investigated by dual in situ hybridisation. PC1 mRNA was localised in prepro-orexin mRNA expressing neurons in the lateral hypothalamus, which suggests a functional relationship.

Animals↗
PubMed2003

Sensitization: a mechanism for melatonin action in the pars tuberalis.

Sensitization of adenylate cyclase is a recently discovered phenomenon. Melatonin can induce a sensitized response of adenylate cyclase in ovine pars tuberalis cells where the receptor for melatonin is endogenously expressed. Although the mechanism is not fully understood, sensitization of adenylate cyclase may be an important part of the mechanism by which melatonin encodes daylength in the pars tuberalis of sheep and other animals. We used this as a hypothesis to search for a natural ligand that would activate adenylate cyclase in ovine pars tuberalis cells. The approach revealed pituitary adenylate cyclase activating polypeptide to be an indirect activator of adenylate cyclase in the ovine pars tuberalis. We discuss this in relation to the mechanism and importance of sensitization to the function to the pars tuberalis.

Adenylyl Cyclases↗
PubMed2003

Photoperiodic programming of body weight through the neuroendocrine hypothalamus.

The photoperiodic mammal undergoes quite remarkable changes in physiology as part of its natural adaptations to seasonal fluctuations in the environment. Changes in energy balance and body weight are among these adaptations. In some seasonal mammals, such as the Siberian hamster (Phodopus sungorus), these changes in body weight have been explored in detail, and there is evidence for tightly controlled systems of energy balance that are coordinated by photoperiod acting via the temporal pattern of melatonin secretion from the pineal gland. The pathways and systems involved appear to be quite distinct from the hypothalamic pathways identified to regulate energy balance in studies of both mice and rats thus far. Instead it appears that in the Siberian hamster a tightly regulated system under the control of photoperiod is able to reset the tone of the systems involved in energy balance regulation. Understanding how photoperiod and melatonin act within the hypothalamus to regulate energy balance offers potentially fundamental and important new insights into the control of energy balance. This review describes the current state of our knowledge.

Animals↗
PubMed2002

Evidence for the biosynthesis of a prolactin-releasing factor from the ovine pars tuberalis, which is distinct from thyrotropin-releasing hormone.

This study demonstrates the presence of two prolactin-releasing (PR) factors in media conditioned by primary pars tuberalis cells prepared from dispersed pars tuberalis tissue. One factor was identified as thyrotropin-releasing hormone (TRH) on the basis of immunoreactivity and following purification by high-performance liquid chromatography and mass spectrometry. The origin of TRH in the pars tuberalis conditioned media was investigated by measuring the expression of glutaminyl-cyclase (QC) by in situ hybridization. QC expression was not detected in pars tuberalis-specific cells, but was relatively abundant in cells in the pars distalis and hypothalamic paraventricular nucleus. These data suggest that TRH is not synthesized by the ovine pars tuberalis and more likely originated from the hypothalamic neuronal processes from the paraventricular nucleus that terminate in the median eminence. The second component of the conditioned media PR bioactivity was insensitive to the TRH-antiserum, less than 1 kDa and was not retained by the C18 reverse-phase column. The biosynthesis of the PR bioactivity by pars tuberalis cells was investigated using cycloheximide, forskolin and melatonin. Cycloheximide reduced the level of PR bioactivity produced by the pars tuberalis cells. Melatonin inhibited the increased level of PR bioactivity stimulated by forskolin. Collectively, these data demonstrate the synthesis of at least one regulator of prolactin secretion by ovine pars tuberalis-specific cells.

Animals↗
PubMed2002

Comparison between medical students' experience, confidence and competence.

OBJECTIVES: This study was undertaken to determine whether or not breadth of clinical experience and student levels of confidence were indicators of competency on standardized simulator performance-based assessments. METHODS: All students (n=144) attending an educational session were asked to complete a 25-point questionnaire regarding specific clinical experiences and levels of confidence in their ability to manage patient problems. For enumeration of clinical experiences, students were asked to estimate the number of times a situation had been encountered or a skill had been performed. For level of confidence, each response was based on a 5-point Likert scale where 1=novice and 5=expert. Students then participated in a standardized simulated performance test. Median and range were calculated and data analysed using Spearman rank correlations. A P-value <0.05 was considered significant. Level of confidence data were compared to performance during clinical rotation and to marks in the anaesthesia final examination. RESULTS: A total of 144 students attended the session, completed the questionnaire and participated in the standardized test. There were wide ranges of experience and confidence in the 25 listed items. Analysis of data showed good correlation between clinical experience and level of confidence. There was no correlation between clinical experience, level of confidence and performance in a standardized simulation test. Neither was there any correlation between level of confidence and clinical grades or written examination marks. CONCLUSIONS: Clinical experience and level of confidence have no predictive value in performance assessments when using standardized anaesthesia simulation scenarios.

Canada↗
PubMed2001

Serine residues 110 and 114 are required for agonist binding but not antagonist binding to the melatonin MT(1) receptor.

Site-directed mutation of serine 110 (Ser(3.35)) and serine 114 (Ser(3.39)) in the human melatonin MT(1) receptor to alanine residues reduced ligand binding affinities of seven known melatonin receptor agonists and partial agonists by 3- to 15-fold. These mutants also displayed a relative reduction in their affinities for melatonin-mediated functional responses of 30- and 14-fold, respectively. In contrast to the observed effects of the agonists and partial agonists, the melatonin receptor antagonist luzindole was found to bind to mutants Ser(3.35)Ala and Ser(3.39)Ala with affinities equivalent to that determined for the wild-type melatonin MT(1) receptor. Luzindole was subsequently confirmed as an antagonist of melatonin-mediated functional responses for both mutant receptors. These studies have identified that in the human melatonin MT(1) receptor, Ser(3.35) and Ser(3.39), in transmembrane domain 3, are critical for the formation of the high-affinity ligand binding site for agonists and partial agonists but not for the antagonist luzindole.

Acetamides↗
PubMed2001

Validity and reliability of undergraduate performance assessments in an anesthesia simulator.

PURPOSE: To examine the validity and reliability of performance assessment of undergraduate students using the anesthesia simulator as an evaluation tool. METHODS: After ethics approval and informed consent, 135 final year medical students and 5 elective students participated in a videotaped simulator scenario with a Link-Med Patient Simulator (CAE-Link Corporation). Scenarios were based on published educational objectives of the undergraduate curriculum in anesthesia at the University of Toronto. During the simulator sessions, faculty followed a script guiding student interaction with the mannequin. Two faculty independently viewed and evaluated each videotaped performance with a 25-point criterion-based checklist. Means and standard deviations of simulator-based marks were determined and compared with clinical and written evaluations received during the rotation. Internal consistency of the evaluation protocol was determined using inter-item and item-total correlations and correlations of specific simulator items to existing methods of evaluation. RESULTS: Mean reliability estimates for single and average paired assessments were 0.77 and 0.86 respectively. Means of simulator scores were low and there was minimal correlation between the checklist and clinical marks (r = 0.13), checklist and written marks (r = 0.19) and clinical and written marks (r = 0.23). Inter-item and item-total correlations varied widely and correlation between simulator items and existing evaluation tools was low. CONCLUSIONS: Simulator checklist scoring demonstrated acceptable reliability. Low correlation between different methods of evaluation may reflect reliability problems with the written and clinical marks, or that different aspects are being tested. The performance assessment demonstrated low internal consistency and further work is required.

Anesthesiology↗
PubMed2001

Photoperiodic regulation of prolactin gene expression in the Syrian hamster by a pars tuberalis-derived factor.

Syrian hamsters exhibit a marked seasonal variation in prolactin secretion. The aim of this study was to analyse the nature of the photoperiodic regulation of prolactin gene expression, and to define the role of melatonin and the pars tuberalis of the anterior pituitary in this process. Pituitary prolactin gene expression, restricted to the pars distalis, was increased in hamsters maintained in long daylengths (16 h : 8 h, light : dark) compared to hamsters exposed to short daylengths (8 h : 16 h, light : dark) for 8-12 weeks. Analysis of single cells by in situ hybridization showed that photoperiod had no effect on the percentage of pars distalis cells expressing prolactin mRNA, but shifted the frequency distribution of prolactin mRNA expression per cell, such that in long photoperiods a greater proportion of cells were recruited to a higher expressing population. In vitro coculture of hamster pars tuberalis fragments increased prolactin promoter-driven luciferase activity in stably transfected GH3 cells in a dose- and duration-dependent manner. Conditioned medium from hamster and ovine pars tuberalis also activated the prolactin promoter. Furthermore, basal and forskolin-stimulated conditioned medium from hamster pars tuberalis increased prolactin mRNA expression in primary cultures of pars distalis cells. Melatonin attenuated the activity of pars tuberalis-conditioned medium but had no direct effect on either prolactin mRNA expression or secretion in pars distalis cell cultures. Finally, pars tuberalis fragments from long photoperiod hamsters stimulated prolactin gene promoter activity to a greater extent than those from short photoperiod hamsters. In conclusion, this study provides the first evidence in a seasonal mammal that the synthesis of prolactin depends on photoperiodic modulation of a pars tuberalis-derived factor. Our data support further the hypothesis that seasonal modulation of prolactin gene expression depends upon a melatonin-dependent paracrine action of the pars tuberalis on pars distalis lactotrophic cells.

Animals↗
PubMed2001

The differential regulation of CART gene expression in a pituitary cell line and primary cell cultures of ovine pars tuberalis cells.

The cocaine-amphetamine regulated transcript (CART) encodes for a protein which has an important role in the regulation of appetite and body weight. To date, no details of the molecular events and signal transduction pathways which regulate this gene are available. We report the identification of CART gene expression in the GH3 pituitary cell line. We have used activators of the cAMP or protein kinase C (PKC) signal transduction pathways to show that, in GH3 cells, CART is transcriptionally up-regulated by activators of the cAMP signal transduction pathway. We also identify CART gene expression in ovine pars tuberalis (PT) tissue and primary cell cultures. In PT cells in contrast to GH3 cells, CART gene expression is upregulated by activators of the PKC signal transduction pathway. Cultured cells have provided a valuable resource for the detailed analysis of specific regulatory mechanisms underlying transcriptional or translational regulation of genes, signal transduction events and many other cellular processes. GH3 and PT cells may therefore provide a resource for the further detailed molecular analysis of the events regulating CART gene expression and processing.

Animals↗
PubMed2001

Localization of the melatonin-related receptor in the rodent brain and peripheral tissues.

Previous studies have provided a limited examination of the expression of the orphan melatonin-related receptor in the pituitary and hypothalamus of human and sheep and retinal tissue in the sheep. The present study reports evidence of conservation of expression in regions of the hypothalamus (dorsal medial hypothalamus, lateral hypothalamus, arcuate nucleus), the epithelial layer lining the third ventricle and the paraventricular thalamic nucleus of the mouse, rat and hamster. An extensive and detailed analysis of melatonin-related receptor mRNA expression in the mouse central nervous system and peripheral tissues is presented. Mapping the distribution throughout the entire mouse brain has revealed new sites of expression in a number of brain nuclei, including preoptic areas, parabrachial nuclei and widespread distribution in the olfactory bulb. Reverse transcriptase-polymerase chain reaction was performed with RNA isolated from peripheral tissues revealing expression of the melatonin-related receptor mRNA in the mouse kidney, adrenal gland, intestine, stomach, heart, lung, skin, testis and ovary. These results suggest a conserved function in neuroendocrine regulation and a potential role in coordinating physiological responses in the central nervous system and peripheral tissues.

Animals↗
PubMed2001

The regulation of body weight: lessons from the seasonal animal.

The hypothalamus is a major regulatory centre involved in the control of many important physiological axes. One of these axes is the regulation of ingestive behaviour. Recent work using a combination of genetic-mutant mouse models together with targeted gene deletions has contributed much to our understanding of how neural pathways of the hypothalamus are involved in the regulation of energy balance in animals. These pathways are also relevant to human energy homeostasis. as mutations in key genes are correlated with obesity. Many of the genes identified mediate the effects of leptin, and are therefore primarily involved in sensing and responding to peripheral signals. In seasonal animals, such as the Siberian hamster (Phodopus sungorus), there is evidence for a higher level of regulation. The systems involved regulate body weight around an apparent 'set-point' through the action of photoperiod via the neurohormone, melatonin. The ability to manipulate energy balance through photoperiod (and melatonin) in the seasonal-animal model offers novel opportunities to identify further fundamental aspects of the control mechanisms involved in the central control of energy homeostasis and body weight.

Animals↗
PubMed2001

The effects of an increase of central blood volume before spinal anesthesia for cesarean delivery: a qualitative systematic review.

UNLABELLED: We evaluated in this qualitative systematic review the efficacy of increasing central blood volume on the incidence of hypotension after spinal anesthesia for elective cesarean delivery. Randomized controlled trials investigating any method of increasing central blood volume before the initiation of obstetric spinal anesthesia were sought by using MEDLINE (1966-2000), Embase (January 1988-April 2000), and the Cochrane Library (Issue 1, 2000). Additional reports from retrieved and review articles, hand searching of non-MEDLINE journals, and abstracts of major anesthesia meetings (1994-1999) were located. The primary outcome was the incidence of hypotension. Secondary outcomes included: ephedrine use, Apgar scores, umbilical cord pH values, and maternal nausea and vomiting. Twenty-three articles met our inclusion criteria with the use of crystalloid preload, colloid preload, and mechanical methods of increasing central volume. Crystalloid preload was inconsistent in preventing hypotension, whereas colloid appeared to be effective in all but one study. Leg wrapping and thromboembolic stockings decreased the incidence of hypotension compared with leg elevation or control. Few differences in fetal outcomes or maternal nausea and vomiting were reported. Increasing central blood volume by using colloid and leg wrapping decreases but does not abolish the incidence of hypotension before spinal anesthesia for elective cesarean delivery. IMPLICATIONS: We performed a systematic review to determine whether fluid loading reduced the incidence of low blood pressure after spinal anesthesia for cesarean delivery. Although no technique totally eliminates the occurrence of hypotension, colloid administration (starch or gelatin containing fluids) and leg wrapping were the most effective.

Adult↗
PubMed2001

A comparison of global ratings and checklist scores from an undergraduate assessment using an anesthesia simulator.

PURPOSE: To determine the correlation between global ratings and criterion-based checklist scores, and inter-rater reliability of global ratings and criterion-based checklist scores, in a performance assessment using an anesthesia simulator. METHOD: All final-year medical students at the University of Toronto were invited to work through a 15-minute faculty-facilitated scenario using an anesthesia simulator. Students' performances were videotaped and analyzed by two faculty using a 25-point criterion-based checklist and a five-point global rating of competency (1 = clear failure, 5 = superior performance). Correlations between global ratings and checklist scores, as well as specific performance competencies (knowledge, technical skills, and judgment), were determined. Checklist and global scores were converted to percentages; means of the two marks were compared. Mean reliability of a single rater for both checklist and global ratings was determined. RESULTS: The correlation between checklist and global ratings was.74. Mean ratings of both checklist and global scores were low (58.67, SD = 14.96, and 57.08, SD = 24.27, respectively); these differences were not statistically significant. For a single rater, the mean reliability score across rater pairs for checklist scores was.77 (range.58-.93). Mean reliability score across rater pairs for global ratings was.62 (.40-.77). Global ratings correlated more highly with technical skills and judgment (r =.51 and r =.53, respectively) than with knowledge. (r =.24) CONCLUSION: Inter-rater reliability was higher for checklist scores than for global ratings; however, global ratings demonstrated acceptable inter-rater reliability and may be useful for competency assessment in performance assessments using simulators.

Anesthesia↗
PubMed2001

Decoding photoperiodic time and melatonin in mammals: what can we learn from the pars tuberalis?

The cellular and molecular mechanisms through which the melatonin signal is decoded to drive/synchronize photoperiodic responses remain unclear. Much of our current understanding of the processes involved in this readout derives from studies of melatonin action in the pars tuberalis of the anterior pituitary. Here, the authors review current knowledge and highlight critical gaps in our present understanding.

Animals↗
PubMed2001

Seasonally inappropriate body weight induced by food restriction: effect on hypothalamic gene expression in male Siberian hamsters.

Male Siberian hamsters undergo physiological weight change in changing photoperiod. Weight loss was induced by food restriction in long days to mimic short-day weight loss, or by food restriction superimposed on short-day weight loss, to test the hypothesis that the hypothalamus differentiates between weight change induced by imposed negative energy balance (inappropriate body weight) and seasonal, appropriate, body weight change, even when these are of similar magnitude. Short-day weight loss was accompanied by reduced POMC and leptin receptor (OB-Rb) mRNA in the arcuate nucleus but elevated cocaine- and amphetamine-regulated transcript. Melanocortin 3-receptor gene expression was reduced in the arcuate nucleus but elevated in the ventromedial nucleus compared with ad libitum-fed long-day controls. Weight loss in long-day restricted animals generated a gene expression profile typical of negative energy balance with low cocaine- and amphetamine-regulated transcript mRNA and elevated OB-Rb. Melanocortin 3-receptor mRNA levels were indistinguishable in short-day and long-day food-restricted hamsters. The hypothalamic correlates of food restriction in short days included up-regulated anabolic neuropeptides and increased OB-Rb mRNA. Low plasma leptin is integrated differently in short-day and long-day restricted animals, and seasonally-inappropriate body weight in either photoperiod engages the compensatory neuropeptide systems involved in the defense of body weight.

Animals↗
PubMed2000

Chimeric melatonin mt1 and melatonin-related receptors. Identification of domains and residues participating in ligand binding and receptor activation of the melatonin mt1 receptor.

Melatonin receptors bind and become activated by melatonin. The melatonin-related receptor, despite sharing considerable amino acid sequence identity with melatonin receptors, does not bind melatonin and is currently an orphan G protein-coupled receptor. To investigate the structure and function of both receptors, we engineered a series of 14 chimeric receptor constructs, allowing us to determine the relative contribution of each transmembrane domain to ligand binding and receptor function. Results identified that when sequences encoding transmembrane domains 1, 2, 3, 5, or 7 of the melatonin mt(1) receptor were replaced by the corresponding domains of the melatonin-related receptor, the resultant chimeric receptors all displayed specific 2-[(125)I]iodomelatonin binding. Replacement of sequences incorporating transmembrane domains 4 or 6, however, resulted in chimeric receptors that displayed no detectable 2-[(125)I]iodomelatonin binding. The subsequent testing of a "reverse" chimeric receptor in which sequences encoding transmembrane domains 4 and 6 of the melatonin-related receptor were replaced by the corresponding melatonin mt(1) receptor sequences identified specific 2-[(125)I]iodomelatonin binding and melatonin-mediated modulation of cyclic AMP levels. To further investigate these findings, site-directed mutagenesis was performed on residues within transmembrane domain 6 of the melatonin mt(1) receptor. This identified Gly(258) (Gly(6.55)) as a critical residue required for high affinity ligand binding and receptor function.

Animals↗