Ras-Erk signaling induces phosphorylation of human TLE1 and downregulates its repressor function.

Signaling mediated by the Ras-extracellular signal-regulated kinase (Erk) pathway often leads to the phosphorylation of transcriptional regulators, thereby modulating their activity and causing concerted changes in gene expression. In Drosophila, the induction of multiple Ras-Erk pathway target genes depends on prior phosphorylation of the general co-repressor Groucho, a modification that downregulates its repressive function. Here, we show that TLE1, one of the four human Groucho orthologs, is similarly phosphorylated in response to Ras-Erk pathway activation, and that this modification attenuates its capacity to repress transcription. Specifically, unphosphorylated TLE1 dominantly suppresses the induction of Ras-Erk pathway target genes in cultured human cells, and the expression of an unphosphorylatable TLE1 derivative causes severe phenotypes in a transgenic Drosophila model system, whereas a phosphomimetic variant of TLE1 exerts only negligible effects. We present data indicating that TLE1 is rapidly excluded from the nucleus following epidermal growth factor receptor pathway activation, an effect that likely accounts for its inability to mediate effective repression under such conditions. Significantly, we find that unphosphorylated TLE1 blocks oncogenic phenotypes induced by mutated H-Ras in human mammary cells, both in vitro and following their implantation in mice. Collectively, our data strongly indicate that phosphorylation of TLE family members and the consequent downregulation of their repressor function is a key conserved step in the transcriptional responses to Ras-Erk signaling, and possibly a critical event in the tumorigenic effects caused by excessive Ras-Erk pathway activity.

Exertional muscle pain in familial Mediterranean fever patients evaluated by MRI and 31P magnetic resonance spectroscopy.

AIM: To evaluate the effect of physical activity on the structural, morphological, and metabolic characteristics of the gastrocnemius muscle in familial Mediterranean fever (FMF) patients, utilizing quantitative (31)P magnetic resonance spectroscopy (MRS), in order to elucidate the mechanism of their exertional leg pain. MATERIALS AND METHODS: Eleven FMF patients suffering from exertional leg pain (eight male, three female; mean age 33 years) and six healthy individuals (three male, three female; mean age 39 years) constituted the control group. All of the participants underwent magnetic resonance imaging (MRI) and non-selective (31)P MRS (3 T) of the leg muscles before and after graded exercise on a treadmill. Phosphocreatine (PCr):inorganic phosphate (Pi), PCr:adenosine triphosphate (ATP) ratios and the intracellular pH of the leg muscles were measured using (31)P MRS. RESULTS: For both groups, normal muscle mass with no signal alterations was observed on the MRI images after exercise. The normal range of pre- and post- exercise MRS muscle parameters was observed in both groups. However, the intracellular pH post-exercise, was significantly higher (less acidic) in the FMF group compared to the control group [pH (FMF) = 7.03 ± 0.02; pH (control) 7.00 ± 0.02; p < 0.0006]. CONCLUSIONS: The finding of a less prominent, post-exercise acidification of the gastrocnemius muscle in this FMF patient group suggests a forme fruste of glycogenosis. This preliminary observation should be further investigated in a future, larger-scale study.

In vivo molecular imaging of met tyrosine kinase growth factor receptor activity in normal organs and breast tumors.

Molecular imaging techniques allow visualization of specific gene products and their physiological processes in living tissues. In this study, we present a new approach for molecular imaging of endogenous tyrosine kinase receptor activity. Met and its ligand hepatocyte growth factor scatter factor (HGF/SF), which mediate mitogenicity, tumorigenicity, and angiogenesis, were used as a model. HGF/SF and Met play a significant role in the pathogenesis and biology of a wide variety of human epithelial cancers and, therefore, may serve as potential targets for cancer prognosis and therapy. We have shown previously that in vitro activation of Met by HGF/SF increases oxygen consumption. In this study, we demonstrate that Met activation in vivo by HGF/SF alters the hemodynamics of normal and malignant Met-expressing tissues. Tumor-bearing BALB/C mice were i.v. injected with HGF/SF and imaged using magnetic resonance imaging (MRI) and Doppler ultrasound. Organs and tumors expressing high levels of Met showed the most substantial alteration in blood oxygenation levels as measured by blood oxygenation level depended (BOLD)-MRI. No significant alteration was observed in tumors or organs that does not express Met. In the liver, which expresses high levels of Met, MRI signal alteration of about 60% was observed. In the kidneys, signal alteration was approximately 30%, and no change was observed in muscles. The extent of MRI signal alteration was also in correlation with HGF/SF doses. Injection of 7 and 170 ng/g body weight resulted in signal alteration of 5% and 30%, respectively, in tumors. Doppler ultrasound measurements demonstrated that these MRI changes are at least partially attributable to altered blood flow. These hemodynamic alterations, measured by MRI and Doppler ultrasound, were used in this study for the molecular imaging of Met activity in vivo. This novel molecular imaging technique may be used for in vivo diagnosis, prognosis, and therapy of Met-expressing tumors.

Vacuolating megalencephalic leukoencephalopathy in 12 Israeli patients.

Leukodystrophy with macrocephaly as the main features of infantile neurodegenerative disease are characteristics of Canavan's disease, L-2-hydroxyglutaric aciduria, type I glutaric aciduria, and Alexander's disease. Also occasionally described are occidental congenital muscular dystrophy, G(M)2-gangliosidosis, metachromatic leukodystrophy, Krabbe's disease, and mucopolysaccharidosis. Since 1995, over 60 patients with a new syndrome, vacuolating megalencephalic leukoencephalopathy, have been described. The syndrome is characterized by macrocephaly, a slowly progressive clinical course of ataxia, spastic paraparesis, and seizure disorder with relatively spared cognition. Unlike other leukodystrophies with macrocephaly (except Alexander's disease), no metabolic marker has been found. We describe a similar group of 12 patients from two different Jewish ethnic origins in whom consanguinity is prominent. These patients have neuroimaging features and magnetic resonance spectroscopy findings indicating that there is an initial increase in white-matter edema with subsequent cystic formation. Consistent with loss of tissue in these areas, brain metabolites are reduced. The familial incidence in this group of patients is suggestive of autosomal-recessive inheritance.

A descriptive study of stress management in a group of pediatric oncology nurses.

Pediatric oncology nursing is associated with highly stressful and emotional situations. This article describes and discusses major sources of occupational stress among a group of nurses participating in a stress management group. The stress sources for these nurses were preoccupation with death and dying, the professional image of the oncology nurse, the nurse as fighter in the war against disease and death, the nurses' perceived isolation from the medical staff, the nurses' perceived inferior professional status compared with that of physicians, emotional overinvolvement with patients and families, suppression of anger, and difficulties in balancing work and home demands. The following factors are suggested as major contributors to the nurses' stress and burnout: increased tendency for irrational-dysfunctional thinking styles (mainly "demandingness" and "awfulizing"), diffuse boundaries between nurses and patients, low professional self-efficacy, and wide prevalence of military metaphors.

Negative association between reported life events and cardiovascular disease risk factors in employed men: the CORDIS Study. Cardiovascular Occupational Risk Factors Determination in Israel.

There is evidence of a link between stressful life events and risk of cardiovascular disease, but the pathway has not been fully explored. The present study of 1859 employed men tested the association between reported intensity of life events and blood pressure and serum lipid levels, risk behaviors, and psychological distress symptoms. The findings revealed a striking disparity in the outcomes. Life events were negatively associated with systolic (p = 0.001) and diastolic (p = 0.038) blood pressure, triglycerides (p = 0.011), and uric acid (p = 0.05), even after controlling for job strain and other possible confounders. In contrast, life events were positively associated with somatic complaints (p < 0.0001), anxiety (p < 0.0001), irritability (p < 0.0001), and depression (p < 0.0001). In addition there was a linear trend between intensity level of life events and low exercise (p = 0.006), smoking (p = 0.007), and alcoholic intake (p = 0.035). The possibility that the above disparity is a product of powerful biases, such as repressive coping and negative affectivity disposition, is discussed.

Role of nuclear magnetic resonance spectroscopy (MRS) in cancer diagnosis and treatment: 31P, 23Na, and 1H MRS studies of three models of pancreatic cancer.

The role of nuclear magnetic resonance spectroscopy (MRS) in pancreatic cancer diagnosis and its treatment were assessed in three models of pancreatic neoplasms. Perfused MIA PaCa-2 human pancreatic cancer cells, s.c. implanted pancreatic tumors in hamsters, and pancreatic tumors induced in situ in rats by direct application of the carcinogen 7,12-dimethyl benzanthracene, were studied by phosphorous ((31)P), sodium ((23)Na), and proton ((1)H) MRS. (31)P spectra of pancreatic cancer were qualitatively similar to those of intact organs. There were, however, variations in peak intensities and ratios. Phosphomonoester signals were prominent in both normal pancreases and tumors, but their levels depended on the proliferation rate and on environmental conditions. Thus, the phosphomonoester:beta-nucleoside triphosphate ratio was 1.15 +/- 0.32 in 90% confluency and 1.31 +/- 0.43 in 70% confluency, and this ratio increased upon lowering the perfusion rate. Total (intra- and extracellular) sodium concentrations, measured in the solid tumors, were 39-40 micromol/g wet weight in normal pancreases. Contrary to a previous hypothesis that malignant transformation is associated with increased sodium content, our (23)Na MRS data showed that there were no significant differences between pancreatic tumors and intact organs. Proton spectra of perchloric acid extracts revealed several differences between tumors and control pancreases. The principal findings were elevated levels of the amino acid taurine, from 1.17 +/- 0.39 micromol/g wet weight in healthy pancreases, to 2.79 +/- 0.71 micromol/g wet weight in pancreatic carcinoma in rats, and lactate levels that increased from 0.92 +/- 0.2 to 6.19 +/- 1.93 micromol/g wet weight, respectively. On the other hand, creatine and glutamate were higher in the normal pancreases. Pancreatic cancer is usually resistant to chemotherapy, and we evaluated the effects of the metabolic inhibitors 2-deoxyglucose and lonidamine on the human pancreatic cancer cells by MRS and cytotoxicity studies. The IC50 of Adriamycin and 2-deoxyglucose were 1.49 +/- 0.18 x 10(6) and 136 +/- 17 microg/ml, respectively. These results were similar to data obtained previously in multidrug-resistant human breast cancer cells, which were highly resistant (33-fold) to Adriamycin but were more susceptible (9-fold) to 2-deoxyglucose than their parental cells.

Health responsibility and workplace health promotion among women: early detection of cancer.

The importance of health responsibility as one aspect of a health-promoting lifestyle has been emphasized repeatedly. Yet there are only a few empirical studies of its role in preventive behavior. We examined the relationship between health responsibility and early-detection practices for breast and cervical cancer. A group of 253 women employees of a large industrial company participated in a cancer screening program subsidized by the employer. They completed questionnaires assessing health responsibility and reported early-detection practices: frequency of breast self-examination and physician breast examinations, frequency of Pap tests, and time lapsed since last Pap test and breast examinations. Health responsibility was a significant independent predictor of breast examination indicators but not of Pap tests. Education level was an important predictor for Pap tests, and age predicted most early-detection practices. The findings lend some support to the role of health responsibility in initiating breast examinations. Better prediction of early-detection practices could be achieved by adding cognitive and emotional components to the existing responsibility scale and by distinguishing between retrospective and prospective responsibility.

Blood lipids and lipoproteins in married and formerly married women.

As part of the Cardiovascular Occupational Risk Factors Determination in Israel (CORDIS) study, the association between marriage termination (divorce/separation or widowhood) and blood lipids and lipoproteins was examined in a sample of 351 healthy women employed in industry. Eighty-seven former spouses were each matched with three married women (N = 264) for age, number of children, smoking status, and type of job (blue/white collar). After controlling for age, number of cigarettes per day, leisure sport participation, and daily coffee consumption, former spouses in the younger women group (younger than 45 years) had significantly higher total cholesterol, total cholesterol ratio, and LDL levels than married women. Among older women (> or = 45 years), there were no significant differences. Significantly more younger former spouses had abnormally high cholesterol and LDL levels. In both age groups, former spouses smoked more cigarettes daily. These differences between the marital status groups may be explained by stress effects and changes in primary prevention practices. If replicated, such findings would delineate a population in need of intervention to reduce disease risk.

Burnout and risk factors for cardiovascular diseases.

The burnout syndrome denotes a constellation of physical fatigue, emotional exhaustion, and cognitive weariness resulting from chronic stress. Although it overlaps considerably with chronic fatigue as defined in internal medicine, its links with physical illness have not been systematically investigated. This exploratory study, conducted among 104 male workers free from cardiovascular disease (CVD), tested the association between burnout and two of its common concomitants--tension and listlessness--and cardiovascular risk factors. After ruling out five possible confounders (age, relative weight, smoking, alcohol use, and sports activity), the authors found that scores on burnout plus tension (tense-burnout) were associated with somatic complaints, cholesterol, glucose, triglycerides, uric acid, and, marginally, with ECG abnormalities. Workers scoring high on tense-burnout also had a significantly higher low density lipoprotein (LDL) level. Conversely, scores on burnout plus listlessness were significantly associated with glucose and negatively with diastolic blood pressure. The findings warrant further study of burnout as a predictor of cardiovascular morbidity and mortality.

Mechanical, biochemical, and structural effects of vitamin D deficiency on the chick heart.

The effects of vitamin D deprivation on the chick heart were investigated from three aspects: cardiac contractility (+/- dP/dT), intracellular high-energy phosphorus compounds, and structural differences. Four-week-old vitamin D-deficient chicks were divided into four groups: Group A served as the normal group and received subcutaneous injections of cholecalciferol; Groups B and C were vitamin D-deficient hearts but perfused differently; Group D received daily subcutaneous injections of 5 micrograms of 1,25(OH)2D3. When the isolated spontaneously beating hearts (modified Langendorff preparation) were perfused with Krebs-Henseleit (KH) solution containing a calcium concentration of 2.5mM, the myocardial contractility of the vitamin D-deficient hearts was significantly increased when compared with group A. After the isolated heart had beaten for one hour, the myocardial contractility in the vitamin D-deficient hearts was found to decline to significantly lower values. Presacrifice administration of 1,25(OH)2D3 improved cardiac performance. Vitamin D deficiency resulted in an enhanced rate of decline of the intracellular high-energy phosphorus compounds. No differences were found in the microscopic study. These observations suggest that vitamin D has a role in cardiac function.

The protective effect of enriched branched chain amino acid formulation in the ischemic heart: a phosphorous-31 nuclear magnetic resonance study.

Branched chain amino acids (BCAA) have been found to have a protective effect on the ischemic myocardium. Isolated rat hearts were perfused with phosphate-free Krebs-Henseleit (KH) solution with or without BCAA. A Latex balloon-tipped catheter was inserted into the left ventricle to measure intracavitary pressures. Hearts were subjected to 18 minutes of 'no flow' global ischemia and then reperfused for 30 mins at 37 degrees C. Metabolism of high energy phosphates during ischemia and recovery was studied by P-31 NMR. Intracellular pH was calculated from the chemical shift of Pi. Pressure recovery was better with KH + BCAA (89 +/- 16%) than with KH (41 +/- 26%) (P = 0.0001); dP/dt recovery was also improved with BCAA (84 +/- 19% vs 27 +/- 27% for KH) (P = 0.0003). After 18 mins of ischemia, ATP levels in the BCAA group were higher than in the KH perfused hearts (33 +/- 20 vs 17 +/- 10% of pre-ischemic value) (P = 0.02). No significant difference was found in the intracellular pH at the end of the ischemic period. Following reperfusion the recovery of pH was better in the BCAA group (7.09 +/- 0.06 vs 7.04 +/- 0.06) (P = 0.03). These results show that BCAA protect the heart from myocardial ischemic injury, decrease depletion of ATP during ischemia, and enhance post-ischemic hemodynamic function.

Acute pancreatitis in rats: a 31P nuclear magnetic resonance study.

High resolution 31P nuclear magnetic resonance (NMR) was used to evaluate the severity of acute pancreatitis in rats. Experimental pancreatitis was induced by intraparenchymal injection of 10% sodium taurocholate. pancreases were removed at various time periods and the NMR spectrum of the whole organ was recorded. Metabolic changes taking place during the progression of the disease were measured and correlated with the pathologic changes. Gradual depletion of the high energy compounds, adenosine triphosphate and phosphocreatine, was observed. The NMR spectral changes paralleled the extension of the pathologic lesions and were found to constitute a reliable indicator of the severity of acute pancreatitis. It is suggested that high resolution NMR may be used to evaluate the pathogenesis and therapy of various forms of experimental pancreatitis.

Smoking and drinking as psychological tools in stressful social situations: assumptions of fiction writers.

The paper reports a study of one potential source of information concerning use and effectiveness of alcohol and cigarettes, which may facilitate habit formation through social learning. A total of 250 episodes from contemporary books of fiction involving either smoking or drinking were analyzed according to various situational and psychological variables characterizing them, including the context, motives preceding consumption, and effects. It was found that smoking and drinking occurred most often in social situations in which intense negative emotions were aroused. The behavioral and emotional consequences of consumption were usually positive. It is suggested that writers tend to attribute to smoking and drinking two main functions: facilitation of skilled performance (either social, cognitive, or technical) and service as psychological tools for coping with emotionally stressful situations. Smoking and drinking occurring in stressful situations, furthermore, are often portrayed as acts of drug prescription, either by the self or by other people.