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A F Junod

Publications and source records attributed to A F Junod.

At least 37 records · Page 2Linked to original sources
PubMed1997

Hyperoxia, unlike phorbol ester, induces glutathione peroxidase through a protein kinase C-independent mechanism.

Human selenium-dependent glutathione peroxidase (GP) is implicated as a mechanism of resistance against oxygen free radicals. The 5' flanking sequence upstream from the coding region of GP contained an oxygen-responsive element termed ORE1 that is responsive to hypoxia, as well as several copies of the activator protein-1 (AP-1)- and AP-1-like-binding sites. In this study, we sought to define the molecular events that lead to GP gene transcription in response to hyperoxia in human umbilical-vein endothelial cells, and asked whether such induction is mimicked and sustained by activation of protein kinase C (PKC) by phorbol esters. Treatment of cells with 100 nM phorbol 12,13-dibutyrate (PdBu) induced a delayed (24-48 h) but significant (2-fold) increase in steady-state GP mRNA levels. Steady-state GP mRNA levels also rose after exposure to 95% O2, again after considerable delay (48-72 h). For both PdBu and oxygen, induction was transcriptionally regulated, as demonstrated by nuclear run-on experiments. The simulations by PdBu and oxygen were additive. In contrast with PdBu, hyperoxia did not stimulate translocation of PKC from the cytosol to the particulate fraction, although the specific activity of both cytosolic and particulate-associated PKC was increased 2-fold in cells exposed to 95% O2 for 5 days. In addition, gel mobility-shift assays using double-stranded tumour-promoting-agent-responsive element (TRE) and nuclear extracts derived from phorbol- and oxygen-treated cells revealed that PdBu, but not hyperoxia, increased AP-1 DNA-binding activity. On the other hand, the up-regulation of GP expression by oxygen could not be accounted for by the ORE1 core sequence, since no specific protein-DNA binding activity could be detected using nuclear extracts from hyperoxic cells and ORE1. Taken together, these results suggest that there may be different molecular mechanisms controlling GP expression. After exposure to PdBu, GP undergoes transcriptional activation via a process that can be readily explained by a classic AP-1 interaction with the TRE sites in the GP promoter. During hyperoxia, GP also undergoes transcriptional activity, but via a process that appears to involve neither TRE nor ORE1.

Binding Sites↗
PubMed1997

Risk factors for a medically inappropriate admission to a Department of Internal Medicine.

OBJECTIVE: To identify patient- and admission-related risk factors for a medically inappropriate admission to a department of internal medicine. METHODS: Cross-sectional study of a systematic sample of 500 admissions to the department of internal medicine of an urban teaching hospital. The appropriateness of each admission and reasons for inappropriate admissions were assessed using the Appropriateness Evaluation Protocol. Risk factors included the time (day of week and holidays) and manner (through emergency department or direct admission) of admission, patient age and sex, health status of patient and spouse, living arrangements, formal home care services, and informal support from family or friends. RESULTS: Overall, 76 (15.2%) hospital admissions were rated as medically inappropriate by the Appropriateness Evaluation Protocol. In multivariate analysis, the likelihood of an inappropriate admission was increased by better physical functioning of the patient (odds ratio [OR], 1.5; 95% confidence interval [CI], 1.1-2.1 [for 1 SD in Physical Functioning scores]), lower mental health status of the patient's spouse (OR, 2.6; 95% CI, 1.3-5.6), receipt of informal help from family or friends (OR, 3.3; 95% CI, 1.5-7.2), and hospitalization by one's physician (OR, 3.6; 95% CI, 1.7-7.5). Receiving formal adult home care was not associated with inappropriateness of hospitalization. CONCLUSIONS: Inappropriate admissions to internal medicine wards are determined by a mix of factors, including the patient's health and social environment. In addition, the private practitioners' discretionary ability to hospitalize their patients directly may also favor medically inappropriate admissions.

Adolescent↗
PubMed1996

Role of oxygen free radicals in cancer development.

In aerobic life, oxidative stress arises from both endogenous and exogenous sources. Despite antioxidant defence mechanisms, cell damage from oxygen free radicals (OFR) is ubiquitous. OFR-related lesions that do not cause cell death can stimulate the development of cancer. This review discusses the effects of oxidative stress at the different stages of carcinogenesis. Mutagenesis through oxidative DNA damage is widely hypothesised to be a frequent event in the normal human cell. A large body of evidence suggests important roles of OFR in the expansion of tumour clones and the acquisition of malignant properties. In view of these facts, OFR may be considered as an important class of carcinogens. Therefore, the ineffectiveness of preventive antioxidant treatments, as documented in several recent clinical trials, is surprising. However, the difficulties of antioxidant intervention are explained by the complexity of both free radical chemistry and cancer development. Thus, reducing the avoidable endogenous and exogenous causes of oxidative stress is, for the present, the safest option. In the near future, new insights in the action of tumour suppressor genes and the DNA repair mechanisms may lead the way to additional tools against carcinogenesis from OFR.

Antioxidants↗
PubMed1996

Detecting acute thoracic aortic dissection in the emergency department: time constraints and choice of the optimal diagnostic test.

STUDY OBJECTIVES: To compare diagnostic strategies for the emergency assessment of patients with suspected acute thoracic aortic dissection and to measure the effect of delays related to the availability of these tests on the selection of the most appropriate one. METHODS: We carried out a decision analysis representing the risks of performing one or two sequential tests, the tests' accuracy, the risks and benefits of treatment, and the time-dependent mortality rate in untreated patients with dissection (1%/hour). Data were drawn from a Medline search. Our subjects were patients who presented to the emergency department with chest pain in whom acute thoracic aortic dissection was suspected. For different clinical probabilities of aortic dissection, we compared the risks and benefits of testing using the following procedures (alone and in combinations): aortography, computed tomography (CT), magnetic resonance imaging (MRI), and both transesophageal (TEE) and transthoracic echocardiography (TTE). We then measured the effect of delays in these tests on the selection of the appropriate procedure. The outcome studied was 30-day survival. RESULTS: We determined that the "threshold" clinical probability of aortic dissection above which the benefits of testing outweigh its risks is low. It ranges from 2% with the most reliable procedure (MRI) to 9% with the least (TTE). At low probability of dissection (< 15%), the accuracy of all tests except TTE is sufficient to rule out dissection. Delays have negligible effect on these results. When the likelihood of dissection is higher, the preferred option is to order a second diagnostic test if the results of the first are negative. The threshold probabilities above which to order a second test range from 15% (CT, then aortography) to 35% (MRI, then aortography). Excessive delays may affect the selection of tests when the likelihood of dissection is high (eg, 50%). Thus, although it is less accurate, a CT scan obtained within 2 hours or a TEE obtained within 6 hours of presentation to the ED yields a higher survival rate than an MRI obtained within 9 hours. Similarly, the benefits of ordering a second test, if the result of the first are negative, outweigh the risks only if the delay in obtaining the test does not exceed 10 hours. CONCLUSION: All patients in whom aortic dissection is suspected, even if the index of suspicion is very low, should undergo one of the available diagnostic procedures (except TTE). A patient with a moderate to high probability of disease should undergo a second investigation if the findings of the first are negative. When the probability of dissection is high, the physician must consider delays in obtaining specific diagnostic tests and order those that will be the most quickly available.

Aortic Dissection↗
PubMed1995

Differential regulation of glutathione peroxidase by selenomethionine and hyperoxia in endothelial cells.

We have studied the effect of selenomethionine (SeMet) and hyperoxia on the expression of glutathione peroxidase (GP) in human umbilical vein endothelial cells. Incubation of HUVEC with 1 x 10(-6) M SeMet for 24 h and 48 h caused a 65% and 86% increase in GP activity respectively. The same treatment did not result in significant changes in GP gene transcription and mRNA levels. Pactamycin, a specific inhibitor of the initiation step of translation, prevented the rise in GP activity induced by SeMet and caused an increase in GP mRNA in both cells grown in normal and SeMet-supplemented medium. Interestingly, SeMet supplementation stimulated the recruitment of GP mRNA from an untranslatable pool on to polyribosomes, so that the concentration of GP mRNA in polyribosomal translatable pools was 50% higher in cells grown in SeMet-supplemented medium than in cells grown in normal medium. On the other hand, cells exposed to 95% O2 for 3 days in normal medium showed a 60%, 394% and 81% increase in GP gene transcription rate, mRNA levels and activity respectively. Hyperoxia also stabilized GP mRNA. Hyperoxic cells grown in SeMet-supplemented medium did not show any change in GP gene transcription and mRNA levels, but expressed an 81% and 100% increase in GP activity and amount of GP mRNA associated with polyribosomes respectively, when compared with hyperoxic cells maintained in normal medium. Thus, GP appeared to be regulated post-transcriptionally, most probably co-translationally, in response to selenium availability, and transcriptionally and post-transcriptionally in response to oxygen.

Cells, Cultured↗
PubMed1995

Effects of oxidative stress and Ca2+ agonists on molecular chaperones in human umbilical vein endothelial cells.

Endothelial cell dysfunction is a key factor in oxidative stress-related pathology. Disruption of Ca2+ homeostasis is thought to be responsible for much of the endothelial cell dysfunction in oxidative stress. The expression of molecular chaperones (MC), which stabilize protein structures in normal and in stress conditions, reflects the Ca(2+)-dependent and -independent stress effects in the different cell compartments. By two-dimensional (2-D) gel electrophoresis combined with immunoblotting or microsequencing, we have identified 12 major MC in human umbilical vein endothelial cells (HUVEC): (i) the endoplasmic reticulum-located MC GRP78, GRP94, protein disulfide isomerase, and calreticulin; (ii) the mitochondrial MC HSP65 and GRP75; and (iii) the cytosolic/nuclear MC HSP27, HSC70, HSP70, HSP90, cyclophilin, and ubiquitin. To differentiate oxidative stress- and Ca(2+)-mediated effects, HUVEC were exposed to 1) xanthine oxidase plus hypoxanthine to generate oxidative stress, 2) ionomycin plus ethylene glycol-bis(beta-aminoethylether)-N,N,N', N'-tetraacetic acid (EGTA) to deplete intracellular Ca2+ stores, or 3) thrombin to increase cytosolic Ca2+. De novo protein synthesis after exposure was quantified by the incorporation of [35S]methionine. Image processing with the MELANIE system was used to create and compare the 2-D maps of [35S]methionine-labeled proteins under conditions 1)-3) with those of the controls. In a total of 24 2-D gels, 9 different MC were detected in at least 5 out 6 experimental replicates and were subjected to numeric analysis. The statistics showed a > 10% increase in GRP78 (p < 0.05), HSP27, cyclophilin, and ubiquitin after oxidative stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium↗
PubMed1995

Differential effects of superoxide, hydrogen peroxide, and hydroxyl radical on intracellular calcium in human endothelial cells.

Changes in intracellular Ca2+ homeostasis are thought to contribute to cell dysfunction in oxidative stress. The hypoxanthine-xanthine oxidase system (X-XO) mobilizes Ca2+ from intracellular stores and induces a marked rise in cytosolic calcium in different cell types. To identify the reactive O2 species involved in the disruption of calcium homeostasis by X-XO, we studied the effect of X-XO on [Ca2+]i by spectrofluorimetry with fura-2 in human umbilical vein endothelial cells (HUVEC). The [Ca2+]i response to X-XO was essentially diminished by superoxide dismutase (SOD) (200 U/ml) and catalase (CAT) (200 U/ml), which scavenge the superoxide anion, O2-, or H2O2, respectively. The [Ca2+]i increase stimulated by 10 nmol H2O2/ml/min, generated from the glucose-glucose oxidase system, or 10 microM H2O2, given as bolus, was about a third of that induced by X-XO (10 nmol O2-/ml/min) but was comparable to that induced by X-XO in the presence of SOD. The X-XO-stimulated [Ca2+]i increase was significantly reduced by 100 microM o-phenanthroline, which inhibits the iron-catalysed formation of the hydroxyl radical. On the other hand, the [Ca2+]i response to low dose X-XO (1 nmol O2-/ml/min) was markedly enhanced in the presence of 1 microM H2O2, which itself had no effect on [Ca2+]i. More than 50% of this synergistic effect was prevented by o-phenanthroline. These results indicate that the effect of X-XO on calcium homeostasis appears to result from an interaction of O2- and H2O2, which could be explained by the formation of the hydroxyl radical.

Calcium↗
PubMed1995

Effects of hypoxanthine-xanthine oxidase on Ca2+ stores and protein synthesis in human endothelial cells.

We have investigated the effects of reactive O2 metabolites generated by the hypoxanthine-xanthine oxidase (HX-XO) system on intracellular Ca2+ and its relation with protein synthesis in human umbilical vein endothelial cells (HUVECs). Spectrofluorometry with fura 2 showed that the oxidative stress induced a rapid transient rise in cytosolic [Ca2+], followed by a sustained elevation above the baseline value. In the presence of La3+, which blocks Ca2+ influx from the extracellular medium, a transient [Ca2+] increase was still observed, but the sustained rise was suppressed. The HX-XO-related [Ca2+] changes were completely prevented by pretreatment with thapsigargin, which depletes intracellular Ca2+ stores. Hence, the effects of HX-XO on Ca2+ homeostasis were due to mobilization of Ca2+ from the intracellular stores with subsequent influx of extracellular Ca2+. HX-XO mobilized more of sequestered Ca2+ than did thrombin, a receptor agonist that depletes only a part of the intracellular Ca2+ stores (the hormone-sensitive stores). To determine the relevance of the HX-XO-related depletion of Ca2+ stores for cell function, we investigated the role of Ca2+ mobilization in the regulation of protein synthesis. Overall protein synthesis in HUVECs was markedly reduced by thapsigargin, which depletes both hormone-sensitive and -insensitive stores, but was not substantially affected by thrombin. Manipulation of the refilling of the Ca2+ stores via the availability of extracellular Ca2+ significantly influenced the thapsigargin-related and the HX-XO-related inhibition of overall protein synthesis. A corresponding effect of extracellular [Ca2+] was seen in polyribosome distribution profiles, which reflected an inhibition of translation initiation in both treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium↗
PubMed1995

[Abdominal aortic aneurysm discovered in a patient with renal insufficiency. A decision analysis].

Decision analysis is an explicit and quantitative technique which is particularly well suited for medical problems involving multiple complex clinical features. In this article, a decision analysis model was used to measure the risks and benefits between different therapeutic options, for a patient with a 4 cm diameter asymptomatic abdominal aortic aneurysm and concomitant end-stage renal failure. A Markov model was built to stimulate the natural history of abdominal aortic aneurysm, its risk of rupture, and the age-dependent dialysis-related mortality. Despite end-stage renal disease that limits life expectancy and increases the surgical risk, the model suggests that surgical resection of the aneurysm (immediate, or delayed if the diameter exceeds 5 cm in diameter) would result in greater life expectancy compared with therapeutic abstention unless the surgical risk exceeds 8.5% and 22%, respectively. Moreover, the gain in life expectancy between immediate and delayed surgery is not significant, and individual decision-making between these two options should include additional elements representing patients' preference. In conclusion, this model emphasizes the usefulness of decision analysis as a decision aid to medical problem-solving. The Markov simulation, which represents in a realistic manner the natural history of aortic aneurysm, and sensitivity analyses reinforce the credibility of this approach.

Aortic Aneurysm, Abdominal↗
PubMed1994

Applicability of decision analysis to everyday clinical practice: a controlled feasibility trial.

OBJECTIVE: To determine whether decision analysis is applicable to routine management of suspected pulmonary embolism in an emergency care setting. DESIGN: Controlled feasibility trial. SETTING: Emergency center of a university hospital. PATIENTS: Outpatients (n = 84) admitted with clinical and scintigraphic evidence of pulmonary embolism. INTERVENTIONS: Patients were treated either with the usual clinical work-up for pulmonary embolism (control group) or using a decision analysis model with three options: no action: angiography followed by treatment if positive; treatment without angiography. RESULTS: All six senior residents in the decision analysis group agreed to fully participate for the 16 months of the study. Summarizing the decision analysis model in a graph was critical to obtain acceptance from all the physicians. Decision analysis (n = 43) and control (n = 41) patients underwent similar numbers of angiographies. However, angiographies for patients who had intermediate clinical probabilities of pulmonary embolism, between 25 and 75%, were more frequent in the decision analysis group (9/13 = 69%) than in the control group (7/20 = 35%). Agreement between clinical probability and lung-scan result was stronger in the decision analysis group. CONCLUSIONS: Decision analysis was successfully used to manage all patients suspected of having pulmonary embolism admitted to an emergency center during the 16-month trial. There was no insuperable obstacle to acceptance of clinical decision analysis by the physicians. Decision analysis may have resulted in a better discrimination between low and intermediate clinical probabilities of pulmonary embolism.

Adult↗
PubMed1994

[Is non-allergenic bronchial hyperreactivity a good diagnostic test for asthma?].

The measurement of non-allergic bronchial hyperreactivity brings limited information in the diagnosis of asthma, whether for epidemiological purposes or for an individual patient. Its sensitivity and specificity in the general population amount to 46 and 89% respectively. It follows that the positive and negative predictive values are of little use when the prevalence of asthma lies between 1 and 10%. The situation is different, more complex, but not necessarily more favorable when one deals with one given patient. The characteristics of the test may vary as a function of the severity of the disease (concept of spectrum bias). Furthermore, the differential diagnosis will be made for other diseases that can also be associated with bronchial hyperreactivity. The negative and positive predictive values of this test will therefore be sufficiently informative only when the diagnosis is already certain.

Asthma↗
PubMed1993

Satellite PET and lung cancer: a prospective study in surgical patients.

Positron emission tomography (PET) appears to be an innovative method for imaging the proliferative activity of malignant tissue, in particular by means of 18F-labelled fluorodeoxyglucose (FDG). The potential role of PET scanning was investigated in a satellite centre as an adjunct to conventional methods for estimating the likelihood of pulmonary malignancy. Therefore the sensitivity of detection of lung cancer in candidates was determined prior to exploratory or therapeutic thoracotomy by FDG PET imaging. The study involved 36 patients with abnormal chest roentgenogram and suspected lung cancer who were due for thoracotomy. The PET scans were evaluated qualitatively and semiquantitatively. Pulmonary malignancy was found in 31/36 patients and 29 had a focal increase in FDG pulmonary uptake. Benign pulmonary lesions were found in 5/36 patients, three of whom had a negative PET scan. The sensitivity of detection of lung cancer by FDG PET was therefore 93.5%. Bayesian study shows that FDG PET could be the most useful method in a population with a low prevalence of lung cancer. As illustrated by our study, a simple FDG PET scanning protocol in a satellite PET centre could provide adequate clinical information and help in deciding subsequent patient management.

Adenocarcinoma↗
PubMed1993

Variable glutathione levels and expression of antioxidant enzymes in human endothelial cells.

We have investigated the relationship between intracellular glutathione levels and the inducibility of the mRNAs encoding the major antioxidant enzymes Cu,Zn superoxide dismutase (Cu,Zn SOD), catalase (CAT), glutathione peroxidase (GP), and the stress protein heme oxygenase (HO) following exposure of human umbilical vein endothelial cells (HUVEC) to either hypoxanthine-xanthine oxidase or 95% O2. Treatment of HUVEC with 2 and 200 microM buthionine sulfoximine (BSO) for 16 h reduced total glutathione (GSH) levels by 51 and 95%, respectively, whereas treatment with 100 microM diethylmaleate (DEM) for 24 h increased the cellular GSH content by 58%. None of these treatments affected the responsiveness of HUVEC to a subsequent oxidant challenge, in terms of antioxidant enzymes activities and mRNA levels. On the contrary, HO mRNA was significantly induced by both BSO and DEM, as well as by hyperoxia, albeit to a different extent. We conclude that intracellular redox changes do not appear to regulate the expression of the mRNAs encoding Cu,Zn SOD, CAT, and GP. Furthermore, factors other than endogenous thiols may play a role in the control of HO mRNA expression.

Blotting, Northern↗
PubMed1993

Effects of inhibition of catalase and superoxide dismutase activity on antioxidant enzyme mRNA levels.

We examined the effects of inhibition of Cu,Zn superoxide dismutase (Cu,Zn SOD) and catalase (Cat) activities on the steady-state mRNA levels of the three major antioxidant enzymes [Cu,Zn SOD, Cat, and glutathione peroxidase (GP)] in human umbilical vein endothelial cells under normoxia and hyperoxia. Inhibition of Cat activity by aminotriazole was not associated with alteration of the other antioxidant enzymes or with potentiation of cell injury. On the other hand, inhibition of Cu,Zn SOD activity by N-N'-diethyl-dithiocarbamate (DDC) was associated with an increase in Cu,Zn SOD mRNA level and a decrease in Cat and GP mRNA level. The combination of DDC and hyperoxia treatments was associated with an additive effect on Cu,Zn SOD message. We propose that these coordinate mRNA changes might be an adaptation to the oxidative environment. This proposal supports the concept that the intracellular O2 metabolites themselves could be the signals that trigger the antioxidant enzymes gene expression.

Amitrole↗
PubMed1993

Alterations in inspiratory and leg muscle force and recovery pattern after a marathon.

We measured lung mechanics and metabolic parameters, first in a series of 15 marathon (M) and 12 half-marathon (H) runners, before and 2.5 h after the race, in order to assess the running effect on respiratory muscles. Maximal static inspiratory (Pi) and expiratory (Pe) pressures, maximal voluntary ventilation (MVV), blood lactate (LAC), arterialized blood gases (ABG), plasma glucose (GLU), and, with a visual analog scale, subjective tiredness were assessed. Pi decreased in M and H without change in Pe, MVV, or lung mechanics; it was still abnormal 2.5 h after the race and did not correlate with LAC, ABG, training parameters, running times, or subjective tiredness. A second study of six other M compared Pi, Pe, adductor pollicis longus (Ppol), and triceps surae (Ptri) forces, aiming to: 1) assess the reproducibility of the prolonged Pi decrease without Pe change, 2) prove that such a decrease corresponded to a real reduction in inspiratory muscle force and was not due to a poor collaboration of the subjects, 3) correlate Pi decrease with the force loss of a potentially tired leg muscle: the triceps surae. We observed a significant and similar decrease in Pi and Ptri, but no change in Ppol and Pe. Moreover, the high correlation between Pi and Ptri diminutions suggested a similar recovery pattern for inspiratory and leg muscles. In conclusion, the prolonged decrease in Pi after such races is compatible with inspiratory muscle fatigue, which, as well as its recovery, is similar to leg muscle tiredness.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Spontaneous heat shock protein synthesis by alveolar macrophages in interstitial lung disease associated with phagocytosis of eosinophils.

Synthesis of heat shock proteins (HSPs) is induced in all cells and tissues after exposure to elevated temperatures, or a variety of other types of injury, including oxidative injury. We have previously reported that stress proteins are induced in monocytes-macrophages during phagocytosis of red blood cells. Receptor-mediated phagocytosis is associated with activation of the respiratory burst, generation of the lipid mediators of inflammation, and increased production of cytokines. Similar activation events have been described in the alveolar macrophage (AM) during pulmonary fibrosis. We therefore analysed the pattern of proteins synthesized by human AMs recovered by bronchoalveolar lavage (BAL) in interstitial lung disease, both under basal conditions and after in vitro exposure to heat or hydrogen peroxide (H2O2). In two out of the 17 cases studied, we observed a high alveolar eosinophilia (10 and 24%, respectively) and phagocytosis, by the AMs, of eosinophilic material. Whereas exposure to heat or H2O2 induced in all AMs the synthesis of the classical HSPs, in these two cases, we found spontaneous synthesis of HSPs and of a 32 kD oxidation-specific stress protein, haeme oxygenase (HO). Exposure of AM to purified eosinophil-derived proteins, such as major basic protein (MBP), eosinophil peroxidase (EPO), eosinophil-derived neurotoxin (EDN), alone or in combination, did not induce stress protein synthesis, further suggesting that phagocytosis is involved in this induction. Stress protein synthesis by AMs may represent a new cellular marker of pulmonary injury and eosinophilic inflammation, and an autoprotective mechanism against oxidative stress.

Adult↗
PubMed1992

Costovertebral joint dysfunction: another misdiagnosed cause of atypical chest pain.

The diagnostic work-up of atypical chest pain frequently leads to invasive procedures. However, this painful symptomatology can sometimes be of benign origin and respond to simple therapeutic manoeuvres. A number of musculoskeletal conditions such as costovertebral joint dysfunctions should be carefully considered. We report five cases in which patient discomfort and high costs could have been avoided if awareness of these conditions had led to a correct diagnosis upon initial physical examination.

Adult↗