Importance of characteristics and modalities of physical activity and exercise in the management of cardiovascular health in individuals with cardiovascular risk factors: recommendations from the EACPR. Part II.

In a previous paper, as the first of a series of three on the importance of characteristics and modalities of physical activity (PA) and exercise in the management of cardiovascular health within the general population, we concluded that, in the population at large, PA and aerobic exercise capacity clearly are inversely associated with increased cardiovascular disease risk and all-cause and cardiovascular mortality and that a dose­response curve on cardiovascular outcome has been demonstrated in most studies. More and more evidence is accumulated that engaging in regular PA and exercise interventions are essential components for reducing the severity of cardiovascular risk factors, such as obesity and abdominal fat, high BP, metabolic risk factors, and systemic inflammation. However, it is less clear whether and which type of PA and exercise intervention (aerobic exercise, dynamic resistive exercise, or both) or characteristic of exercise (frequency, intensity, time or duration, and volume) would yield more benefit for each separate risk factor. The present paper, therefore, will review and make recommendations for PA and exercise training in the management of cardiovascular health in individuals with cardiovascular risk factors. The guidance offered in this series of papers is aimed at medical doctors, health practitioners, kinesiologists, physiotherapists and exercise physiologists, politicians, public health policy makers, and individual members of the public. Based on previous and the current literature overviews, recommendations from the European Association on Cardiovascular Prevention and Rehabilitation are formulated regarding type, volume, and intensity of PA and regarding appropriate risk evaluation during exercise in individuals with cardiovascular risk factors.

Reduced pH and contractility in failing rat cardiomyocytes.

AIM: To determine whether reduced cardiomyocyte contractility in heart failure is associated with reduced intracellular pH (pH(i)). Involvement of the Na(+)/H(+) exchanger and the H(+)/K(+) ATPase were investigated with specific blockers. METHODS: Myocardial infarction and subsequent heart failure in Sprague-Dawley rats were induced by chronic occlusion of the left coronary artery. 6 weeks post-ligation, contractility (cell shortening) and pH(i) (BCECF fluorescence) were recorded in freshly dissociated cardiomyocytes during 2-10 Hz electrical stimulation, with or without either Na(+)/H(+) exchanger or H(+)/K(+) ATPase inhibition. RESULTS: Elevated end-diastolic and reduced peak systolic pressures confirmed heart failure. Increased heart weights (20-30%; P < or = 0.01) and cardiomyocyte lengths and widths (22-25%; P < or = 0.01) confirmed substantial cardiac hypertrophy. In myocytes isolated from sham operated rats, a positive staircase response occurred with stimulation rates from 2 to 7 Hz; further increases in stimulation rate up to 10 Hz reduced contractility. In contrast, pH(i) fell progressively over the entire stimulation range. In failing myocytes, pH(i) was consistently 0.07 pH units lower and contractility 40% lower (P < or = 0.01) than sham control values; the shape of the contractility staircase remained similar to controls. At all stimulation frequencies, Na(+)/H(+) exchanger inhibition reduced pH(i) by 0.05 pH units (P < or = 0.01) and contractility by 22% (P < or = 0.05) in cardiomyocytes from the heart failure group. A significantly smaller decrease of pH(i) and reduction in contractility was observed after inhibition of Na(+)/H(+) exchanger (10 micro m HOE694) in sham myocytes. H(+)/K(+) ATPase inhibition (100 micro m SCH28080) had no effect on pH(i). CONCLUSION: Reduced pH(i) is accompanied by reduced cardiomyocyte contractility in isolated myocytes from post-MI heart failure. The data suggest compensatory Na(+)/H(+) exchanger activation in heart failure, whereas H(+)/K(+) ATPase does not appear to contribute significantly to pH(i) maintenance.

Trans-sodium crocetinate does not affect oxygen uptake in rats during treadmill running.

Trans-sodium crocetinate (TSC), the isomer of the carotenoid compound crocetin, is found markedly to increase survival in hemorrhagic shock subsequent to 50-60% blood loss, mainly via restored resting oxygen consumption (VO(2)), blood pressure and heart rate. The proposed mechanism is that TSC increases oxygen diffusivity, and thus availability, in plasma. If this were found to be a prominent feature in the oxygen transfer from blood to skeletal muscle fiber mitochondria, increased VO(2) during exercise would be expected because of reduced partial pressure of venous oxygen (increased utilization), which we aimed to elucidate in this study. Male Sprague-Dawley rats were intravenously injected with 0.3 mL kg(-1) TSC (40 microg mL(-1)) or placebo and immediately thereafter tested on a ramped treadmill test protocol. Rats were introduced to the experimental protocols beforehand. Administration of TSC had a neutral effect on submaximal and maximal VO(2) (VO(2max)) as well as running performance measured as maximal running time and maximal aerobic running velocity. Thus, in this study we cannot report any effects of TSC on steady-state submaximal VO(2) or VO(2max) at exhaustive exercise.

Identification and regulation of the gastric H+/K+-ATPase in the rat heart.

UNLABELLED: Previous reports indicate that H+/K+-adenosine triphosphatase (ATPase) might be expressed in the heart. AIMS: The objectives of the present study were to explore the presence of H+/K+-ATPase protein and gene expression in the rat heart and to investigate whether the enzyme could contribute to potassium transport across the sarcolemma. METHODS AND RESULTS: We performed reverse transcription-polymerase chain reaction (RT-PCR) on mRNA from myocardium and isolated cardiomyocytes using primers specific for the gastric H+/K+-ATPase alpha-subunit. The PCR products were sequenced and the predicted gastric H+/K+-ATPase sequence was verified. Western blots from myocardium detected a 34-kDa band and a 94-kDa band, indicating the beta-subunit and alpha-subunit of the gastric H+/K+-ATPase, respectively. Immunocytochemistry detected significant immunoreactivity of the beta-subunit in cardiomyocytes. H+/K+-ATPase-dependent potassium transport was assessed by 86Rb+-uptake in isolated cardiomyocytes. Both ouabain and the selective H+/K+-ATPase inhibitor Schering 28080 reduced 86Rb+-uptake at maximum specific inhibition, by 70 and 25%, respectively; the effects were additive. Competitive RT-PCR analysis indicated a significant upregulation of the myocardial H+/K+-ATPase in heart failure after myocardial infarction. CONCLUSION: The gastric isoform of H+/K+-ATPase is expressed in rat cardiac myocytes, both at transcript and protein levels. Functional studies indicate that the enzyme could contribute to potassium and pHi regulation in cardiomyocytes.

Cardiomyocyte contractility and calcium handling partially recover after early deterioration during post-infarction failure in rat.

The aim of the study was to determine whether progression of heart failure is associated with deterioration of cardiomyocyte function. Cell dimensions, contractility and calcium transients were measured in cardiomyocytes isolated from the left ventricle of female Wistar rats 1, 4, and 13 weeks after coronary artery ligation or sham-operation. Relative cardiomyocyte shortening decreased from 26% in controls to 11% 1 week after myocardial infarction and recovered to 18 and 20% after 4 and 13 weeks, respectively. Diastolic and systolic calcium concentrations increased markedly 1 week after myocardial infarction with subsequent reduction after 4 and 13 weeks. Time to 50% relaxation was prolonged by 31% after 1 week and 20% after 4 and 13 weeks with corresponding changes in diastolic calcium clearance. Cardiomyocyte length increased by 6, 24, and 26% after 1, 4 and 13 weeks, respectively, whereas myocyte width increased by 4, 11 and 27%. Cardiomyocytes adjacent to the infarct hypertrophied more and initially had more markedly impaired function than in the remote area. Left ventricular diastolic diameter assessed by echocardiography increased by 47, 66 and 84% after 1, 4 and 13 weeks, respectively, and systolic diameter increased by 120, 162 and 194%. Left ventricular end-diastolic pressure increased from 6 mmHg to 24, 25 and 36 mmHg. Whereas initial deterioration of cardiac function is associated with reduced cardiomyocyte contractile function, chronic heart failure progression is not accompanied by further impairment of intrinsic cardiomyocyte contractility in this model. Cardiomyocyte hypertrophy and dysfunction are more marked adjacent to the infarction.

Surgical manipulation, but not moderate exercise, is associated with increased cytokine mRNA expression in the rat soleus muscle.

Interleukin (IL)-6 production in contracting skeletal muscle and IL-6 concentration in plasma are increased after prolonged and strenuous exercise. However, as tissue stress or damage are unspecific triggers of increased cytokine levels, we examined whether moderate muscle activity is an independent stimulus for cytokine expression, and to which extent invasive procedures might affect the results. Soleus muscles were isolated from sedentary rats or from rats that had been running on a treadmill at moderate intensity (70% of maximal oxygen uptake) for 1 h. In another group the soleus muscle was prepared in situ and stimulated intermittently at 5 Hz for 1 h, so that maximal developed force declined by 30%. In situ prepared soleus muscles not subjected to electrical stimulation were used as controls. Messenger RNA (mRNA) expression of 11 cytokines was analysed in the soleus muscles using multiprobe RNAse protection assay, and IL-6 plasma concentration was measured by enzyme-linked immunosorbent assay. Treadmill exercise did not affect the mRNA expression of any of the measured cytokines in the soleus muscle. Irrespective of electrical stimulation, mRNA expression of IL-6 and IL-1beta were significantly increased in the surgically manipulated soleus muscles. Interleukin-6 plasma concentration was not affected by treadmill running or electrical stimulation. Conclusion, gentle surgical manipulation is a strong stimulus for IL-6 and IL-1beta mRNA synthesis in skeletal muscle, whereas exercise or electrical muscle stimulation at moderate intensity does not independently affect cytokine mRNA levels in the contracting soleus.

Chronic exposure to carbon monoxide and nicotine: endothelin ET(A) receptor antagonism attenuates carbon monoxide-induced myocardial hypertrophy in rat.

The aims of the present study were to determine the effects of endothelin ET(A) receptor antagonism on carbon monoxide (CO)-induced cardiac hypertrophy and endothelin-1 (ET-1) expression and to compare myocardial effects of chronic nicotine with CO exposure. Female Sprague-Dawley rats (n = 84) were randomized to three groups exposed 20 h/day to CO (200 ppm), nicotine (500 microg/m3), or air for 14 consecutive days. In each exposure group, animals were randomized to ET(A) receptor antagonist LU 135252 in drinking water (0.5 mg/ml) or placebo. Myocardial ET-1 and atrial natriuretic peptide (ANP) expression was measured by competitive RT-PCR and plasma ET-1 by immunoassay. Carboxyhemoglobin was 22.1 +/- 0.3% in CO-exposed animals and 2.8 +/- 0.3% in controls. Plasma nicotine was 57 +/- 7 ng/ml and plasma cotinine was 590 +/- 23 ng/ml in nicotine-exposed animals and below detection levels in controls. CO exposure induced a 21% increase in right ventricular hypertrophy (p < 0.01), a 7% increase in left ventricular hypertrophy (p < 0.01), a 25% increase in right ventricular ET-1 expression (p < 0.05), and an eightfold increase in ANP expression (p = 0.08). ET(A) receptor antagonism reduced right ventricular hypertrophy by 60% (p < 0.05) with no significant effect on left ventricular hypertrophy or myocardial ET-1 expression. Chronic nicotine exposure did not significantly affect cardiac weights or ANP and ET-1 expression. We conclude that ET(A) receptor antagonism reduces right ventricular hypertrophy induced by chronic CO exposure, whereas CO-induced myocardial ET-1 expression remains unchanged.

Atrioventricular plane displacement in female endurance athletes.

INTRODUCTION: A novel hypothesis for increased ventricular pumping describes the heart as a displacement pump, in which atrioventricular plane displacement (AVPD) is an important mechanism. The hypothesis predicts that AVPD increases at high heart rates. The aim of the present study was to determine whether AVPD increases during exercise at high heart rates. A secondary aim was to study the left ventricular function and dimensions in endurance-trained young female athletes. METHODS: Eight female cross-country skiers (18.5 +/- 0.9 yr, 169.3 +/- 2.9 cm, 55.7 +/- 4.2 kg, and 64.8 +/- 3.7 mL x kg(-1) x min(-1) in maximal oxygen uptake) were compared with seven sedentary female controls (18.0 +/- 0.6 yr, 175.0 +/- 2.5 cm, 71.1 +/- 4.1 kg, and 42.8 +/- 3.0 mL x kg(-1) x min(-1) in maximal oxygen uptake). Cardiac anatomy and function were assessed by echocardiography. RESULTS: Whereas left ventricular ejection fraction (LVEF) increased in both groups, AVPD fell significantly during exercise. AVPD did not correlate with LVEF, and there were no differences in AVPD between groups either at rest or during exercise. Cardiac output index (mL x lean body mass(-0.67)) and stroke volume index (mL x lean body mass(-1)) were higher in the trained group. The trained group had a larger left ventricular mass, and left ventricular internal dimensions, scaled to lean body mass. CONCLUSIONS: This first study to examine AVPD at rest and during exercise in endurance-trained athletes did not indicate that AVPD is an important mechanism of increased cardiac pumping during exercise. Endurance-trained female athletes have larger left ventricular dimensions and increased function as compared with sedentary subjects.

Increased contractility and calcium sensitivity in cardiac myocytes isolated from endurance trained rats.

OBJECTIVE: Regular exercise enhances cardiac function and modulates myocyte growth in healthy individuals. The purpose of the present study was to assess contractile function and expression of selected genes associated with intracellular Ca2+ regulation after intensity controlled aerobic endurance training in the rat. METHODS: Female Sprague-Dawley rats were randomly assigned to sedentary control (SED) or treadmill running (TR) 2 h per day, 5 days per week for 2, 4 or 13 weeks. Rats ran 8-min intervals at 85-90% of VO2max separated by 2 min at 50-60%. Myocyte length, intracellular Ca2+ (Fura-2), and intracellular pH (BCECF) were measured in dissociated cells in response to electrical stimulation at a range of stimulation rates. RESULTS: The increase in VO2max plateaued after 6-8 weeks, 60% above SED. After 13 weeks, left and right ventricular weights were 39 and 36% higher than in SED. Left ventricular myocytes were 13% longer, whereas width remained unchanged. After 4 weeks training, myocyte contractility was approximately 20% higher in TR. Peak systolic intracellular Ca2+ and time for the decay from systole were 20-35 and 12-17% lower, respectively. These results suggest that increased myofilament Ca2+ sensitivity is the dominant effect responsible for enhanced myocyte contractility in TR. Intracellular pH progressively decreased as stimulation frequency was increased in the SED group. This decrease was markedly attenuated in TR and the intracellular pH was significantly higher in the TR group at a stimulation rate of 5-10 Hz. This effect may contribute to the increased contractility observed at the higher stimulation frequencies in TR. A higher intrinsic myofilament Ca2+ sensitivity was observed in permeabilised myocytes from the TR group under conditions of constant pH and [Ca2+]. Western blot analysis indicated 21 and 46% higher myocardial SERCA-2 and phospholamban, but unaltered Na+/Ca(2+)-exchanger levels. Competitive RT-PCR revealed that TR significantly increased Na+/H(+)-exchanger mRNA. CONCLUSION: Intensity controlled interval training increases cardiomyocyte contractility. Higher myofilament Ca(2+)-sensitivity, and enhanced Ca(2+)-handling and pH-regulation are putative mechanisms. Our results suggest that physical exercise induces adaptive hypertrophy in cardiac myocytes with improved contractile function.

Attenuated endothelin- mRNA expression with endothelin- receptor blockade during hypoxaemia and reoxygenation in newborn piglets.

UNLABELLED: We investigated the cause of decreased plasma endothelin-1 (ET-1) during hypoxaemia and reoxygenation in newborn piglets subjected to simultaneous blocking of the ET-1 receptors. Changes in plasma ET-1 and prepro-ET-1 mRNA expression in the main pulmonary artery and the left lower lobe in the lung were studied in 1-2-d-old piglets. Ten minutes prior to hypoxaemia, the hypoxaemia group (n = 10) was given saline, two groups (both n = 9) were given 1 and 5 mg/kg i.v. SB 217242 (an ET-1 receptor antagonist). Two groups served as normoxic controls, with and without SB 217242 5 mg/kg i.v. Hypoxaemia was induced by ventilating with 8% O2 until base excess was <-20 mmol/l or mean arterial blood pressure was <20 mmHg. Reoxygenation was performed for 2 h with room air. During hypoxaemia, plasma ET-1 decreased in the hypoxaemia group, remained unchanged in the 1-mg group and increased in the 5-mg group. At the end of reoxygenation, plasma ET-1 was above baseline in the 1-mg and 5-mg groups. In the pulmonary artery, the hypoxaemia group showed 2- to 5-fold higher prepro-ET- 1 mRNA expression compared to all the other groups (p < 0.05). There were trends for higher prepro-ET-1 mRNA expression in pulmonary tissue in the hypoxaemia group compared to the two receptor-blocking groups (p < 0.07). CONCLUSIONS: We conclude that hypoxaemia and reoxygenation increase prepro-ET-1 mRNA expression in the pulmonary artery in newborn piglets. These observations suggest that the half-life of ET-1 is decreased during hypoxaemia and reoxygenation in newborn piglets.

Attenuated endothelin-1 mRNA expression with endothelin-1 receptor blockade during hypoxaemia and reoxygenation in newborn piglets.

We investigated the cause of decreased plasma endothelin-1 (ET-1) during hypoxaemia and reoxygenation in newborn piglets subjected to simultaneous blocking of the ET-1 receptors. Changes in plasma ET-1 and prepro-ET-1 mRNA expression in the main pulmonary artery and the left lower lobe in the lung were studied in 1-2-d-old piglets. Ten minutes prior to hypoxaemia, the hypoxaemia group (n = 10) was given saline, two groups (both n = 9) were given 1 and 5 mg/kg i.v. SB 217242 (an ET-1 receptor antagonist). Two groups served as normoxic controls, with and without SB 217242 5 mg/kg i.v. Hypoxaemia was induced by ventilating with 8% O2 until base excess was 20mmol/l or mean arterial blood pressure was < 20mmHg. Reoxygenation was performed for 2h with room air. During hypoxaemia, plasma ET-1 decreased in the hypoxaemia group, remained unchanged in the 1-mg group and increased in the 5-mg group. At the end of reoxygenation, plasma ET-1 was above baseline in the 1-mg and 5-mg groups. In the pulmonary artery, the hypoxaemia group showed 2- to 5-fold higher prepro-ET-1 mRNA expression compared to all the other groups (p < 0.05). There were trends for higher prepro-ET-1 mRNA expression in pulmonary tissue in the hypoxaemia group compared to the two receptor-blocking groups (p < 0.07). CONCLUSIONS: We conclude that hypoxaemia and reoxygenation increase prepro-ET-1 mRNA expression in the pulmonary artery in newborn piglets. These observations suggest that the half-life of ET-1 is decreased during hypoxaemia and reoxygenation in newborn piglets.

Chronic carbon monoxide exposure in vivo induces myocardial endothelin-1 expression and hypertrophy in rat.

Smoking is associated with endothelial dysfunction and increased plasma levels of endothelin-1. The component of tobacco smoke inducing these effects is unknown. Carbon monoxide induces hypoxia, and there is evidence of carbon monoxide acting as a local mediator in both endothelial and smooth muscle cells. The purpose of this study was to determine whether chronic carbon monoxide exposure similar to that experienced by smokers affects myocardial endothelin-1 expression. Sprague-Dawley female rats were exposed to carbon monoxide 100 ppm for one week or to 100 ppm for one week and 200 ppm for a second week. Carboxyhaemoglobin was 12+/-0.9% in the low and 23+/-1.1% in the high carbon monoxide exposure group. Endothelin-1 expression was measured by competitive reverse transcriptase polymerase chain reaction. High carbon monoxide exposure increased endothelin-1 mRNA by 54+/-12% (P<0.001) in the left ventricle and by 53+/-12% (P<0.001) in the right ventricle. In the low carbon monoxide exposure group corresponding changes were 43+/-14% (P=0.06) and 12+/-16%(P=0.29). Right ventricular weight increased by 18+/-7% (P=0.02) after high and by 16+/-5% (P=0.02) after low exposure. Left ventricular weight was elevated by 5+/-2% (P=0.05) when both exposure groups were compared to controls. We conclude that chronic carbon monoxide exposure leading to carboxyhaemoglobin levels similar to those observed in smokers increases endothelin-1 gene expression and induces myocardial hypertrophy in the rat.

Expression of calcium channels in adult cardiac myocytes is regulated by calcium.

In addition to playing a significant role in cardiac excitation-contraction coupling, intracellular Ca2+ ([Ca2+]i) can regulate gene expression. While the mechanisms regulating expression of Ca2+ channels are not entirely defined, some evidence exists for Ca2+-dependent regulation. Using an adult ventricular myocyte culture system, we determined the effects of Ca2+ on: (1) abundance of mRNA for L-type Ca2+ channel alpha1 subunit (DHP receptor); (2) amount of DHP receptors; and (3) whole-cell Ca2+ current (ICa). Rat ventricular myocytes were cultured for 1-3 days in serum-free medium containing either normal (1.8 mM) or high (4.8 mM) Ca2+. Exposing myocytes to high Ca2+ rapidly elevated [Ca2+]i as determined by fura-2. Northern blot analysis revealed that culturing cells in high Ca2+ produced 1.5-fold increase in mRNA levels for the DHP receptor. The abundance of DHP receptors, determined by ligand binding, was two-fold greater in myocytes after 3 days in high Ca2+. Moreover, peak ICa was larger in myocytes cultured for 3 days in high Ca2+ (-17.8+/-1.5 pA/pF, n=26) than in control cells (-11.0+/-1.0 pA/pF, n=23). Voltage-dependent activation and inactivation, rates of current decay, as well as percent increases in ICa elicited by Bay K8644 were similar in all groups. Therefore, larger ICa is likely to represent a greater number of functional channels with unchanged kinetics. Our data support the conclusion that transient changes in [Ca2+]i can modulate DHP receptor mRNA and protein abundance, producing a corresponding change in functional Ca2+ channels in adult ventricular myocytes.

Na,K-pump concentration in hypertrophied human hearts.

Several studies have associated myocardial dysfunction with reduced myocardial Na,K-pump concentration, but whether impaired Na,K-pump capacity is a pathogenetic factor or an epiphenomenon related to accompanying cardiac hypertrophy is not established. We measured Na,K-pump concentrations in 10 hypertrophied and 11 normal weighted hearts obtained at autopsy using [3H]ouabain as ligand. Specific [3H] ouabain binding site concentration (OBC) in the left ventricle (LV) averaged 449 +/- 40 (pmol.g-1 wet weight; mean +/- SEM) in hypertrophied and 598 +/- 36 in normal weighted ventricles (P = 0.02). A trend towards lower LV OBC (-19%; P = 0.25) was found in hypertrophied hearts from patients with congestive heart failure as compared with non-failing hypertrophied hearts. In multivariate analysis with 18 variables including age and heart failure, only LV weight correlated independently with LV OBC (r = -0.61; P = 0.003). When OBC was related to either dry weight or to protein content, a 25-35% reduction was consistently found in hypertrophied LV, whereas RV OBC was similar in both groups. In conclusion, myocardial Na,K-pump concentration and thus the capacity to maintain homeostasis is reduced in LV, but not in RV, of hypertrophied hearts. Whether the moderately reduced myocardial Na,K-pump concentration is a pathogenetic factor in LV dysfunction remains to be determined.

Continual electric field stimulation preserves contractile function of adult ventricular myocytes in primary culture.

To model with greater fidelity the electromechanical function of freshly isolated heart muscle cells in primary culture, we describe a technique for the continual electrical stimulation of adult myocytes at physiological frequencies for several days. A reusable plastic cover was constructed to fit standard, disposable 175-cm2 tissue culture flasks and to hold parallel graphite electrodes along the long axis of each flask, which treated a uniform electric field that resulted in a capture efficiency of ventricular myocytes of 75-80%. Computer-controlled amplifiers were designed to be capable of driving a number of flasks concurrently, each containing up to 4 x 10(6) myocytes, over a range of stimulation frequencies (from 0.1 to 7.0 Hz) with reversal of electrode polarity after each stimulus to prevent the development of pH gradients around each electrode. Unlike quiescent, unstimulated myocytes, the amplitude of contraction, and velocities of shortening and relaxation did not change in myocytes paced at 3-5 Hz for up to 72 h. The maintenance of normal contractile function in paced myocytes required mechanical contraction per se, since paced myocytes that remained quiescent due to the inclusion of 2.5 microM verapamil in the culture medium for 48 h also exhibited a decline in contractility when paced after verapamil removal. Similarly, pacing increased peak calcium current compared with quiescent cells that had not been paced. Thus myocyte contraction at physiological frequencies induced by continual uniform electric field stimulation in short-term primary culture in defining medium maintains some biophysical parameters of myocyte phenotype that are similar to those observed in freshly isolated adult ventricular myocytes.

Mortality and causes of death in a 10-year follow-up of patients treated for self-poisonings in Oslo.

The present 10-year follow-up study includes all patients (N = 926; 50% females) treated in the medical departments in Oslo for self-poisonings during one year (1980). Seventeen percent were considered suicidal attempts upon admission, 25% among the non-substance abusers and 8% among the abusers. At follow-up, 207 patients (22%) were dead (62% males). The mortality rate was highest among the abusers. The most common causes of death were suicide (21%), heart disease (17%), opiate abuse (15%), and accidents/wounds (13%). Forty-one percent of the suicides occurred during the first two years of the follow-up period. The suicides were by poisoning (57%), hanging (20%), and other methods (23%). The female mortality rate decreased in the second half of the follow-up period whereas the male rate did not change. The risk of death within 10 years after discharge increased with age and was higher in men and in abusers, whereas social group and motive for suicide were not predictive factors. The females had an excess suicide rate of 182 (36-327, 95% CI) in the first year after the self-poisoning and 61 (36-87, 95% CI) in the total period. The corresponding figures for males were 70 (19-122) and 21 (12-30). The only factor associated with an increased suicide rate was a suicidal motive upon the admission for self-poisoning with a 3.1 (1.7-5.8, 95% CI) times increased risk of suicide in the 10-year follow-up period.

Adult rat ventricular myocytes cultured in defined medium: phenotype and electromechanical function.

We studied primary short-term cultures of adult rat ventricular myocytes in defined medium to determine whether phenotype and electromechanical function are maintained in rod-shaped, quiescent cells. Although > 80% of the myocytes retained their rod-shaped in vivo morphology for up to 72 h, contractile function as measured by cell edge motion declined 30-50% from 6 to 24 h, paralleling a 68% shortening of action potential duration. From 24 to 72 h, contractility remained unchanged. Ca2+ channel current density increased 55% after 24-48 h and then returned to the level of freshly isolated cells (9 +/- 1 pA/pF, mean +/- SE). Resting membrane potential (-71 +/- 1 mV) and action potential overshoot (34 +/- 3 mV) did not change. The ratio of alpha- to beta-myosin heavy chain mRNA and the level of cardiac alpha-actin mRNA were maintained for 8 days. Thus quiescent adult rat ventricular myocytes in defined medium undergo extensive phenotypic adaptation within 72 h of isolation, despite maintenance of a rod-shaped morphology and stable levels of contractile protein mRNA, which may limit their suitability for electrophysiological and contractile function studies.

Isolation and characterization of human and rat cardiac microvascular endothelial cells.

Although reciprocal intercellular signaling may occur between endocardial or microvascular endothelium and cardiac myocytes, suitable in vitro models have not been well characterized. In this report, we describe the isolation and primary culture of cardiac microvascular endothelial cells (CMEC) from both adult rat and human ventricular tissue. Differential uptake of fluorescently labeled acetylated low-density lipoprotein (Ac-LDL) indicated that primary isolates of rat CMEC were quite homogeneous, unlike primary isolates of human ventricular tissue, which required cell sorting based on Ac-LDL uptake to create endothelial cell-enriched primary cultures. The endothelial phenotype of both primary isolates and postsort subcultured CMEC and their microvascular origin were determined by characteristic histochemical staining for a number of endothelial cell-specific markers, by the absence of cells with fibroblast or pericyte-specific cell surface antigens, and by rapid tube formation on purified basement membrane preparations. Importantly, [3H]-thymidine uptake was increased 2.3-fold in subconfluent rat microvascular endothelial cells 3 days after coculture with adult rat ventricular myocytes because of release of an endothelial cell mitogen(s) into the extracellular matrix, resulting in a 68% increase in cell number compared with CMEC in monoculture. Thus biologically relevant cell-to-cell interactions can be modeled with this in vitro system.

Transcriptional regulation in cardiac muscle. Coordinate expression of Id with a neonatal phenotype during development and following a hypertrophic stimulus in adult rat ventricular myocytes in vitro.

The transcriptional regulatory mechanisms in heart muscle that direct cardiac development and allow for a flexible, adaptive response to physiologic stress are not well understood. We demonstrate that a negative regulator of gene transcription termed Id that has been described predominantly in proliferating cell lines and in undifferentiated tissue during growth, is expressed in freshly isolated terminally differentiated adult rat ventricular myocytes, in contrast to most other tissues in the adult rat. Id mRNA expression is regulated in ventricular myocytes during post-natal development, peaking at the transition from hyperplastic to hypertrophic growth at day 17 in the rat, declining subsequently to lower, stable levels in adult myocytes. Although Id mRNA becomes undetectable in adult ventricular myocytes 48 h following isolation in the absence of serum, it can be rapidly reinduced by an alpha-adrenergic agonist, accompanied by increased protein synthesis and the reexpression, in defined media, of the neonatal genes prepro-ANP and skeletal muscle alpha-actin. Thus, the differential regulation of Id during cardiac development, the presence of Id mRNA in normal cardiac myocytes, and its increased expression following a hypertrophic stimulus all suggest a role for this transcriptional regulator in the control of cardiac muscle cell phenotype.

Calcium-induced net potassium uptake of pig hearts in vivo.

To determine whether the catecholamine-induced myocardial potassium uptake could be mimicked by increasing extracellular and intracellular calcium concentrations in vivo, we measured changes in myocardial potassium balance in nine anaesthetized open-chest pigs with PVC-valinomycin electrodes in arterial and coronary sinus blood. CaCl2 infusion (200-400 mumol min-1) into the left coronary artery increased coronary sinus blood calcium concentration from 2.29 (2.19-2.42) to 4.63 (3.76-5.67) mmol l-1 (median, 95% confidence interval, P = 0.01) indicating a similar increment in myocardial extracellular calcium concentration. The contractility measure LV dP/dt increased 95 (76-147) %, indicating a substantial increment in intracellular calcium concentration. During the CaCl2 infusion coronary sinus potassium concentration declined to a nadir 0.12 (0.09-0.17) mmol l-1 below baseline (P = 0.008) whereas arterial concentration remained unchanged. Peak myocardial potassium uptake was 18 (7-32) mumol min-1 100 g-1 and occurred 150 (110-195) s after start of infusion. The response remained unaltered after adrenoceptor blockade by prazosin and propranolol. Prolonged CaCl2 infusion caused a net myocardial potassium loss which was accompanied by metabolic and haemodynamic indications of myocardial ischaemia. These findings are consistent with enhanced Na-K pump activity in the intact beating pig heart in response to increased extracellular and intracellular calcium concentrations.