Assay of G protein-coupled receptor kinase activity by rhodopsin phosphorylation.
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Animals↗
Biomedical subjects
E Kelly
Publications and source records attributed to E Kelly.
Development of strategies to identify the learning needs of baccalaureate nursing students.
Meeting the learning needs of nursing students today requires a shift in paradigms and the use of multimethods of teaching. This article describes case-control research undertaken to measure the effects of learning strategies on the final grades of first semester junior level, baccalaureate nursing students. The strategies used and how the students learned to use the strategies are presented. Research results did not provide statistical significance because of the small sample size. However, several trends related to ethnicity, student age, and the content of nursing courses were found. These findings suggest the need for further nursing education research in the areas of information processing and the use of learning strategies within academic and clinical settings.
Adult↗
Ultrasonographic guidance for needle biopsy of breast lesions.
The use of imaging-guided needle biopsies of the breast to establish the histology of lesions has become an important tool in the clinical management of breast pathology. Avoiding excision of benign lesions and allowing definitive surgery of malignant lesions to be planned from the outset are to the benefit of the patient and also decrease the cost of breast health care. The use of ultrasonographic guidance for such procedures provides a safe and effective real time technique to sample tissue while the patient lies in a comfortable supine position. The operator who has a good understanding of the procedure will be able to obtain reliable specimens efficiently and consistently.
Biopsy, Needle↗
The effects of pharmacological modulation of KATP on the guinea-pig isolated diaphragm.
The purpose of the present study was to investigate the functional consequences of KATP modulation in the normal and the metabolically inhibited guinea-pig isolated diaphragm using the K+ channel openers cromakalim, pinacidil, RP49356 (N-methyl-2-(3-pyridil)-tetrahydrothiopyran-2-carbothiami de-1-oxide) and ZM260384 (2-(2,2-bis(difluoromethyl)-6-nitro-3,4-dihydro-2H-1,4-benzoxazine -4-yl)pyridine-N-oxide) and the K+ channel inhibitors glibenclamide, phentolamine and ciclazindol. All K+ channel openers accelerated the decline in function induced by intermittent tetanic contractions following metabolic inhibition and delayed the development of contracture. Cromakalim also improved the recovery of twitch tension following 10 min intermittent tetanic stimulation in the hypoxic guinea-pig diaphragm preparation. Of the K+ channel inhibitors tested, only ciclazindol, at the highest concentration tested (10 microM), significantly delayed the decline in tetanic tension following metabolic inhibition in the guinea-pig isolated diaphragm. None of the inhibitors significantly accelerated the development of contracture. All inhibitors however, antagonised the actions of the K+ channel opener, cromakalim. The results indicate that opening of KATP can accelerate the decline in function following metabolic inhibition in the guinea-pig isolated diaphragm. In the absence of K+ channel openers however, KATP does not appear to contribute to this decline under the conditions of the present study.
Adenosine Triphosphate↗
Laparoscopic cholecystectomy in the transplant population.
BACKGROUND: The results of laparoscopic cholecystectomy in a group of transplant recipients were reviewed to determine the safety and efficacy of the procedure in the setting of immunosuppression. METHODS: All solid-organ-transplant recipients who underwent laparoscopic cholecystectomy over a 3-year period were reviewed. Indication for operation, conversion to open procedure, length of stay, and complications were characterized. These results were compared to the registry data of all laparoscopic cholecystectomies performed at the same institution. RESULTS: There were 26 transplant patients who underwent laparoscopic cholecystectomy including renal, heart, double lung, and heart-lung recipients. The mean age was 47 years. Symptomatic cholelithiasis was the most common indication in 73% of patients followed by acute cholecystitis in 11%. Seven patients (27%) underwent conversion to an open procedure. Three patients (11.5%) experienced a minor complication in hospital. Median length of stay was 2.5 days. One patient died during a subsequent unrelated operation. These results compared favorably to the registry experience at the same institution where the mean age was 49 years, 24% of cases were performed for acute cholecystitis, there was a 10% complication rate, median length of stay was 2 days, and 3 deaths occurred in hospital. The only statistically significant difference was a lower conversion rate (11% vs 27%) in the registry vs transplant group. CONCLUSIONS: This experience confirms that laparoscopic cholecystectomy is as safe in the transplant population as the general population. Despite a slightly higher conversion rate to an open procedure, the advantages of short hospital stay, low morbidity, and early return to preoperative routines remain equivalent.
Adult↗
Metallothionein and HSP-72 are induced in the liver by hemorrhagic shock and resuscitation but not by shock alone.
BACKGROUND: Previous reports have indicated that HSP-72 and metallothionein mRNA undergo induction in the liver after resuscitated hemorrhagic shock. In this study we investigated whether unresuscitated shock triggers induction and whether protein induction also occurs. METHODS: Rats were subjected to resuscitated and unresuscitated shock protocols of varying severity; livers were isolated and processed for Northern, Western, and immunohistochemical analysis. Cadmium binding assay was used to measure metallothionein protein. RESULTS: Unresuscitated shock led to no induction of HSP-72 or metallothionein. Severe resuscitated shock led to prompt induction of HSP-72 mRNA and protein in hepatocytes, up to 20-fold over sham group; metallothionein mRNA induction appeared later than HSP-72 and did not lead to elevated protein levels. Mild resuscitated shock had little effect. CONCLUSIONS: These findings indicate resuscitated severe shock, not shock alone, leads to induction of HSP-72 and metallothionein in the liver. Metallothionein expression lags behind HSP-72 expression.
Animals↗
Cytomegalovirus infections in Toronto child-care centers: a prospective study of viral excretion in children and seroconversion among day-care providers.
OBJECTIVES: To prospectively determine the rate of cytomegalovirus shedding in children and the rate of seroconversion to cytomegalovirus in providers at 38 infant-toddler day care centers in Toronto, Canada. METHODS: Urine was collected for shell vial assay in 471 children between the ages of 3 and 42 months. Providers (n = 206) were tested for the presence of cytomegalovirus antibody by latex agglutination. Of the 68 providers who were seronegative, 56 were retested approximately 1 year later. RESULTS: Viruria was documented in 79 (17%) children and antibody in 67% of providers. Seropositivity was significantly related to country of birth outside Canada, presence of children at home < 5 years of age and increased household size. Seroconversion was documented in 12.5% (n = 7). Of these providers 71% worked at centers where workers never wore gloves for diaper changing vs. 33% of those who did not seroconvert (P = 0.06), and all were younger than 30 years vs. 59% of those who did not seroconvert (P = 0.04). In centers with viruria the association of seroconversion with lack of glove use was enhanced (P = 0.04). Seroconversion was marginally more likely in providers working with infants only than with infants and toddlers or with toddlers alone. Logistic regression confirmed that seroprevalence was more likely in providers who were born outside Canada, had children younger than age 5 years in the household and with an increased number of people in the household. Seroconversion was more likely if the provider worked at centers not using gloves for diaper changes, worked with infants only rather than with toddlers and infants and was < 30 years old, with each factor contributing independently to the model. CONCLUSIONS: Cytomegalovirus infection is common in children and providers in Toronto day-care centers.
Adolescent↗
Use of CD14 and lipopolysaccharide binding protein mRNA expression as markers for acute rejection in rat small bowel transplantation.
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Acute-Phase Proteins↗
Inhibition of ADP-ribosyltransferase increases synthesis of Gs alpha in neuroblastoma x glioma hybrid cells and reverses iloprost-dependent heterologous loss of fluoride-sensitive adenylate cyclase.
Exposure of NG108-15 cells to 50 mM nicotinamide [an inhibitor of mono(ADP-ribosyl)transferase] for 18 hr led to an increase in membrane associated Gs alpha measured either as cholera toxin substrate or by immunoblotting with a specific antiserum. Prolonged exposure of NG108-15 cells to iloprost is followed by homologous loss of iloprost sensitivity, and heterologous loss of fluoride-dependent activation of adenylate cyclase. Nicotinamide reversed the loss of fluoride sensitivity, but failed to restore iloprost-dependent activation of adenylate cyclase. These results with nicotinamide in NG108-15 cells contrasted with those from platelets, which also exhibit heterologous desensitization of fluoride sensitivity following prolonged exposure to iloprost. Treatment of platelets with 50 mM nicotinamide for 18 hr led to an increase of 75.0 +/- 19.4% in the amount of membrane associated cholera toxin substrate. However, there was no associated increase in the abundance of Gs alpha as determined by immunoblotting. Furthermore, in platelets there was no restoration by nicotinamide of the iloprost-dependent loss of fluoride-sensitive adenylate cyclase activity. It follows that heterologous desensitization in platelets is accompanied by inactivation of Gs alpha, which is retained within the plasma membrane in its inactive state. The nicotinamide-dependent increase in the abundance of membrane associated cholera toxin substrate and immunoreactive Gs alpha in NG108-15 cells is associated with an increase of 72.0 +/- 20.3% in the levels of mRNA encoding Gs alpha. The capacity of nicotinamide to increase the abundance of membrane associated Gs alpha was reversed when the cells were cultured in the presence of 20 micrograms/mL cycloheximide. These results suggest that the ability of nicotinamide to increase the abundance of Gs alpha in NG108-15 cells is mediated by de novo protein synthesis.
ADP Ribose Transferases↗
Tuberculin sensitivity and the BCG scar in tuberculosis contacts.
SETTING: Tuberculosis clinic, St Vincents Hospital, Dublin. OBJECTIVE: To determine the influence of past BCG vaccination on tuberculin sensitivity in tuberculosis contacts. DESIGN: Prospective cohort study of tuberculin sensitivity using the Mantoux technique (5 TU). RESULTS: In 1050 contacts of 96 cases of tuberculosis, BCG scars were noted in 76% of the contacts. Approximately 50%, 30% and 20% of the contacts respectively had negative (0-4 mm), positive (5-14 mm) and strongly positive (15+ mm) Mantoux responses. The presence of a BCG scar was not found to significantly influence the degree of tuberculin sensitivity. The degree of tuberculin sensitivity increased after 20 years and decreased after 60 years of age. Contacts of sputum-positive cases of tuberculosis had larger Mantoux responses. CONCLUSIONS: The study suggests that age of the contact and sputum status of the index case are important determinants of the degree of tuberculin sensitivity. Attributing a 'positive' Mantoux response to past BCG vaccination may be encouraging a false sense of security in contacts recently exposed to an infectious case of tuberculosis. Contact management guidelines may need to be reviewed and perhaps modified to reflect this finding.
Adolescent↗
Hemorrhagic shock.
A great deal has been learned about the pathophysiologic condition of hemorrhagic shock. The response of the hormonal and inflammatory mediator systems in patients in hemorrhagic shock appears to represent a distinct set of responses different from those of other forms of shock. The classic neuroendocrine response to hemorrhage attempts to maintain perfusion to the heart and brain, often at the expense of other organ systems. This intense vasoconstriction occurs via central mechanisms. The response of the peripheral microcirculation is driven by local tissue hypoperfusion that results in vasodilation in the ischemic tissue bed. Activation of the systemic inflammatory response by hemorrhage and tissue injury is an important component of the pathophysiologic condition of hemorrhagic shock. Activators of this systemic inflammatory response include ischemia/reperfusion injury and neutrophil activation. Capillary "no-flow" with prolonged ischemia and "no-reflow" with reperfusion may initiate neutrophil activation in patients in hemorrhagic shock. The mechanisms that lead to decompensated and irreversible hemorrhagic shock include (1) "arteriolar hyposensitivity" as manifested by progressive arteriolar vasodilation and decreased responsiveness of the microcirculation to alpha-agonists, and (2) cellular injury and activation of both proinflammatory and counterinflammatory mechanisms. These changes represent a failure of the microcirculation. Redistribution of cardiac output and persistent gut ischemia after adequate resuscitation may also contribute to the development of irreversible hemorrhagic shock. Treatment of hemorrhagic shock includes rapid operative resuscitation to limit activation of the mediator systems and abort the microcirculatory changes that result from hemorrhagic shock. Volume resuscitation and control of hemorrhage, should occur simultaneously. The end point in volume resuscitation of hemorrhagic shock must be maintenance of organ system and cellular function. Whether we use adequate urine output, correction of lactic acidemia, optimization of oxygen delivery, or oxygen consumption as our specific goal, the general objective is to provide adequate crystalloid solution and packed red blood cells to achieve and maintain normal organ and cellular perfusion and function.
Adrenal Glands↗
Methodological issues in patient satisfaction surveys.
Examines some of the methodological problems encountered in conducting patient satisfaction surveys, including the sampling frames, quality of survey data and instruments, non-response problems, and reporting and interpretation of results. Proposes guidelines and lays out an agenda for future research.
Data Collection↗
Effects of acute and chronic ethanol on cyclic AMP accumulation in NG108-15 cells: differential dependence of changes on extracellular adenosine.
1. This study investigated the effects of acute and chronic ethanol on basal, agonist- and forskolin-stimulated cyclic AMP formation in NG108-15 mouse neuroblastoma x rat glioma hybrid cells, and examined the role of changes in extracellular adenosine concentrations on the effects observed. 2. NG108-15 cells incubated acutely with ethanol (1-200 mM) displayed concentration-dependent increases in basal and iloprost-stimulated (300 nM; a prostanoid IP receptor agonist) cyclic AMP accumulation but a concentration-dependent decrease in forskolin-stimulated (10 microM) accumulation. 3. Cells treated chronically with ethanol (200 mM) for 48 h displayed increases over control in basal, iloprost- (0.001-10 microM) and forskolin (0.01-100 microM)-stimulated cyclic AMP formation. However, chronic ethanol did not affect [3H]-iloprost binding to cell membranes. 4. Inclusion of adenosine deaminase (ADA; 1 unit ml-1) during the incubation period to measure cyclic AMP accumulation completely abolished the increase in basal accumulation following chronic ethanol, but did not affect the increase in iloprost stimulation. On the other hand ADA partially reversed the increase in forskolin stimulation following chronic ethanol, but even in the presence of high concentrations of ADA (5 units ml-1) the forskolin stimulation remained elevated above control. 5. Cells treated chronically with the adenosine receptor agonist 5'-(N-ethylcarboxamido)-adenosine (NECA; 10 microM for 48 h) displayed a reduction in subsequent NECA- and forskolin-stimulated cyclic AMP accumulation, but iloprost stimulation was not affected. ADA included acutely during the incubation period to measure cyclic AMP accumulation abolished the reduction in forskolin but not NECA stimulation produced by the chronic NECA pretreatment. 6. We have previously noted that ethanol inhibits NG108-15 cell proliferation and alters cell morphology.To mimic this, cells were incubated in the absence of foetal calf serum for 48 h. Following this time, basal, iloprost- and forskolin-stimulated cyclic AMP formation was enhanced over that in cells grown in the presence of serum.7. These results indicate that chronic ethanol enhances cyclic AMP formation in intact NG108-15 cells by more than one mechanism: one involves increased extracellular adenosine concentrations and the other a change in the transduction system beyond the receptor, possibly involving the adenylyl cyclase enzyme. Furthermore the ethanol-induced changes in cyclic AMP accumulation may relate to alterations in NG108-15 cell growth and development.
Adenosine↗
Measurement of adenylyl cyclase activity in cell membranes.
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Adenylyl Cyclases↗
Gs alpha-dependent and -independent desensitisation of prostanoid IP receptor-activated adenylyl cyclase in NG108-15 cells.
NG108-15 mouse neuroblastoma x rat glioma cells were treated with the prostanoid IP receptor agonist iloprost (1 microM) and the time course of changes in the levels of prostanoid IP receptors, adenylyl cyclase activity, and the alpha-subunit of the stimulatory guanine nucleotide binding regulatory protein, Gs, were measured. Incubation of cells with iloprost produced a biphasic time course of desensitisation of prostanoid IP receptor-activated adenylyl cyclase. Parallel analysis of iloprost-induced loss of membrane Gs alpha, NaF-stimulated adenylyl cyclase and [3H]iloprost binding suggested only monophasic curves, with t0.5 values similar to the initial phase of desensitisation of iloprost-stimulated adenylyl cyclase activity. This suggests that the loss of receptor and Gs alpha occur at the same time and account for the initial period of desensitisation due to iloprost pretreatment. Pretreatment of NG108-15 cells with cholera toxin produced a near complete loss of membrane-associated Gs alpha, but the loss of [3H]iloprost binding due to iloprost treatment was not affected by pretreatment with cholera toxin, suggesting that prostanoid IP receptors can be down-regulated in the absence of any coupling to Gs. The second phase of desensitisation of iloprost-stimulated adenylyl cyclase activity, during which there was no further change in NaF-stimulated adenylyl cyclase or in the membrane levels of Gs alpha, was not due to protein kinase A activation, since elevating intracellular cyclic AMP levels with forskolin did not subsequently decrease iloprost-stimulated adenylyl cyclase activity or [3H]iloprost binding. These results demonstrate that iloprost pretreatment of NG108-15 cells induces two distinct phases of desensitisation; an initial desensitisation due to concurrent loss of prostanoid IP receptors and Gs alpha, and then a further desensitisation by an as yet uncharacterized mechanism during which there is no further loss of Gs alpha.
Adenylyl Cyclases↗
Compliance of the respiratory system in newborn infants pre- and postsurfactant replacement therapy.
Surfactant administration causes a rapid and dramatic improvement in gas exchange, but paradoxically, studies have failed to show an improvement in the mechanical properties of the lung. We have measured dynamic and static (passive flow-volume technique) compliance before and after a single dose of bovine lipid extract surfactant in 22 premature infants with RDS. This had no effect on the measured dynamic compliance. In contrast, surfactant significantly increased static compliance from 0.41 +/- 0.02 to 0.55 +/- 0.04 mL/cm H2O/kg. This improvement was the result of a substantial recruitment of lung volume after surfactant administration. This led us to reduce ventilator pressures, which produced an increase in both dynamic and static compliance, but did not recruit additional volume. We conclude that surfactant causes a substantial increase in static compliance due to volume recruitment, which is consistent with reports of increase in the measured FRC. However, despite this improvement, the compliance is still below our normal range.
Female↗
Use of heat shock for culturing Clostridium difficile from rectal swabs.
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Bacteriological Techniques↗
Chronic ethanol reduces immunologically detectable Gq alpha/11 alpha in NG108-15 cells.
Recent work has shown that chronic ethanol treatment inhibits receptor-stimulated phosphoinositide hydrolysis in NG108-15 cells and that ethanol exerts this effect primarily at the level of the guanine-nucleotide binding protein (G protein). Here we investigated the effects of ethanol exposure on the expression of Gq alpha/11 alpha, two highly homologous G protein alpha-subunits that have been implicated as regulators of phosphoinositidase C. Addition of ethanol (10-200 mM) to the culture medium for 48 h caused a concentration-dependent decrease in the immunologically detectable levels of Gq alpha/11 alpha. A small (approximately 15%) reduction in Gq alpha/11 alpha was observed after only 6 h of exposure to 200 mM ethanol, but membrane levels were reduced by 31% at 48 h. The ethanol-induced loss of Gq alpha/11 alpha was apparently independent of factors present in the foetal calf serum component of the culture medium. These results suggests that the decrease in receptor-mediated phosphoinositide hydrolysis following chronic ethanol treatment of NG108-15 cells may be mediated in part by a reduction in the membrane levels of Gq alpha/11 alpha.
Electrophoresis, Polyacrylamide Gel↗