Metabolic and satiating effects and consumer acceptance of a fibre-enriched Leberkas meal: a randomized cross-over trial.

PURPOSE: The Western diet is poor in dietary fibre and previous efforts to increase fibre intake were not successful. The aim of this study was to develop sensorically appealing, fibre-enriched convenience foods. As a showcase, we prepared a fibre-enriched, fat-reduced Leberkas served in a roll and compared the reformulated product with the standard product. METHODS: The design was a randomized, single-blinded cross-over study. A Leberkas meal enriched with 19.2 g of wheat fibre and resistant dextrin as well as fat- and energy-reduced (30% less calories) was served to 20 middle-aged healthy volunteers (10 male, 10 female) and compared to the standard product in a random order. Blood was repeatedly taken over a 4 h period to measure metabolic parameters as well as satiety hormones, such as glucagon-like-peptide 1, cholecystokinin, peptide YY. Satiety and consumer acceptance of the fibre-enriched meal were assessed by visual analogue scales and a questionnaire. RESULTS: The fibre-enriched meal showed very small significant effects at only single time points in postprandial blood glucose (at 120 min, p = 0.050) and glucoseAUC fibre 22,079 ± 2819, standard 22,912 ± 3583 (p = 0.030). The profiles of satiety hormones were comparable between both meals. No differences in subjective satiation, taste and consumer acceptance were observed between the two products, despite a marked reduction in fat and energy content of the reformulated product. CONCLUSION: It is possible to enrich a popular convenience product with dietary fibre and to markedly reduce energy content without loss of sensory qualities or satiety suggesting that development and promotion of healthier convenience foods may be a useful strategy to tackle obesity and other diet-related diseases.

Deep body composition phenotyping during weight cycling: relevance to metabolic efficiency and metabolic risk.

Weight cycling may lead to adverse effects on metabolic efficiency (i.e. adaptive thermogenesis or 'metabolic slowing') and metabolic risks (e.g. increased risk for insulin resistance and the metabolic syndrome). In order to investigate these topics, the partitioning of fat and lean mass (i.e. the change in the proportion of both compartments) needs to be extended to the organ and tissue level because metabolic risk differs between adipose tissue depots and lean mass is metabolically heterogeneous being composed of organs and tissues differing in metabolic rate. Contrary to data obtained with severe weight loss and regain in lean people, weight cycling most likely has no adverse effects on fat distribution and metabolic risk in obese patients. There is even evidence for an increased ability of fat storage in subcutaneous fat depots (at the trunk in men and at the limbs in women) with weight cycling that may provide a certain protection from ectopic lipid deposition and thus explain the preservation of a favourable metabolic profile despite weight regain. On the other hand, the mass-specific metabolic rate of lean mass may increase with weight gain and decrease with weight loss mainly because of an increase and respective decrease in the proportion (and/or activity) of metabolically active organ mass. Obese people could therefore have a higher slope of the regression line between resting energy expenditure (REE) and fat-free mass that leads to an overestimation of metabolic efficiency when applied to normalize REE data after weight loss. Furthermore, in addressing the impact of macronutrient composition of the diet on partitioning of lean and fat mass, and the old controversy about whether a calorie is a calorie, we discuss recent evidence in support of a low glycaemic weight maintenance diet in countering weight regain and challenge this concept for weight loss by proposing the opposite.

Protein normalization in different adipocyte models and dependence on cell size.

Various in vitro models are commonly used to study adipose tissue function. However, methods for protein normalization in dependence of differentiation and fat cell size are poorly defined. Therefore, the aim of this study was to characterize frequently used housekeepers during adipose differentiation in fat cells of varying cell size and between different subjects to allow a reliable and robust data interpretation. Human preadipocytes, SGBS, 3T3-L1, and mature cells were used to study the housekeeper profile. Glyceraldehyde-3-phosphate dehydrogenase was the most stable internal control in human preadipocytes, whereas all others showed substantial variation. In SGBS and 3T3-L1 cells none of the housekeepers revealed stable protein levels during differentiation. In mature adipocytes GAPDH and α-tubulin were found to be suitable as internal loading controls. Importantly, there was no substantial variation between different donors. However, Coomassie-staining turned out to represent an appropriate alternative as a stable and highly sensitive internal standard for all cell models tested. In conclusion, standard housekeeping proteins have substantial limitations in studies of adipogenesis and in dependence of fat cell size. Coomassie-staining turned out to be a sensitive and stable alternative as internal control for western blot studies during adipogenesis. However, inter-subject variability was low for the investigated housekeeper.

LIGHT (TNFSF14) inhibits adipose differentiation without affecting adipocyte metabolism.

OBJECTIVE: The member of the tumor necrosis factor family LIGHT (lymphotoxin-like inducible protein that competes with glycoprotein D for herpesvirus entry on T cells; TNFSF14 (tumor necrosis factor super family protein 14) is primarily expressed in lymphocytes, in which it induces the expression of pro-inflammatory cytokines and alterations of lipid homeostasis. Recently, the protein was shown to be upregulated in obesity and to induce cytokine secretion from adipocytes. RESEARCH METHODS AND PROCEDURES: Using an automated complementary DNA (cDNA) screen, LIGHT was identified to inhibit adipose differentiation. As cellular models for adipogenesis mouse 3T3-L1, human SGBS (Simpson-Golabi-Behmel syndrome) and primary human preadipocytes differentiated in vitro were used as well as primary human adipocytes to study adipocyte functions. Analysis of lipid deposition by Oil Red O staining, mRNA expression by quantitative reverse transcriptase-PCR, nuclear factor (NF)-κB activation as well as protein secretion by enzyme linked immunosorbent assay and Luminex technology was performed. RESULTS: LIGHT was found to inhibit lipid accumulation in the three models of preadipocytes in a dose-dependent manner without cytotoxic effects. This inhibition of differentiation was probably because of interference at early steps of adipogenesis, as early exposure during differentiation showed the strongest effect, as assessed by decreased peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer-binding protein-α (C/EBPα) mRNA expression. In contrast to TNFα, basal and insulin-stimulated glucose uptake and lipolysis of terminally differentiated mature adipocytes were not altered in the presence of LIGHT. At a concentration sufficient to inhibit differentiation, secretion of proinflammatory cytokines was not significantly induced and NF-κB activity was only modestly induced compared with TNFα. CONCLUSION: LIGHT is a novel inhibitor of human adipocyte differentiation without adversely influencing central metabolic pathways in adipocytes.

Expression and secretion of RANTES (CCL5) in human adipocytes in response to immunological stimuli and hypoxia.

Obesity and related disorders represent states of systemic low-grade inflammation. Chemokine secretion by adipocytes may initiate leukocyte infiltration in obese adipose tissue and thus mediate an important step in the establishment of chronic immune activation. The chemokine RANTES (regulated upon activation normal T cell expressed and secreted)/CCL5 is a chemoattractant for various leukocyte subsets. This study was designed to examine whether RANTES is expressed and released by human adipocytes and how its expression is regulated. RANTES expression under basal conditions was studied in mature adipocytes. Cells were therefore challenged with lipopolysaccharide (LPS), interferon (IFN)-gamma, interleukin (IL)-4, monocyte chemoattractant protein (MCP)-1 or exposed to low oxygen pressure. RANTES was expressed and secreted constitutively in most samples of mature adipocytes from the omental and the subcutaneous depot. RANTES release was dependent on adipocyte size and also seemed to be higher from cells of obese donors. Hypoxia (4% O (2)) caused an approximately 36% increase of RANTES release. Human adipocytes express the chemokine RANTES and are thus identified as a novel cellular source of this immune mediator. LPS and IFNgamma do not seem to play a significant role for the expression of RANTES in contrast to moderate hypoxia, which points to a distinct role in the innate immune system.

Effect of conditioned media from mature human adipocytes on insulin-stimulated Akt/PKB phosphorylation in human skeletal muscle cells: role of BMI and fat cell size.

Obesity is associated with chronic low-grade inflammation. It is currently hypothesized that products secreted from fat cells not only cause this proinflammatory condition but may also have a direct impact on the development of insulin resistance in skeletal muscle. The aim of this study was to investigate if adipocyte-conditioned media interfere with insulin-stimulated Akt/PKB phosphorylation in in vitro differentiated human skeletal muscle cells. Primary human skeletal muscle cells were exposed to adipocyte-conditioned media from subjects with a wide range of BMI. Insulin-induced phosphorylation of the signaling proteins Akt/PKB and ERK-2 was analyzed using a bead-based fluorescence detection system and was correlated to BMI and fat cell size. Adipocyte-conditioned media reduced insulin-stimulated Akt/PKB phosphorylation in a manner depending on BMI and fat cell volume. This inhibition in serine phosphorylation was comparable to that observed in TNF-alpha-treated control cultures. Conditioned media from omental adipocytes reduced Akt/PKB phosphorylation moderately to a greater extent compared to media from subcutaneous fat cells from the same donors (p<0.05). Furthermore, there were significant associations between the concentration of selected adipokines and Akt/PKB phosphorylation. These data provide first direct evidence that secreted factors from freshly isolated mature fat cells separated according fat cell size reduce insulin-stimulated Akt/PKB phosphorylation in human skeletal muscle cells and may contribute to the pathogenesis of obesity-associated insulin resistance.

Constitutive and regulated expression and secretion of interferon-gamma-inducible protein 10 (IP-10/CXCL10) in human adipocytes.

OBJECTIVE: Chemokine secretion by adipocytes has been postulated to initiate leukocyte infiltration in adipose tissue and might mediate an important step in the establishment of obesity-related chronic immune activation. The chemokine interferon (IFN)gamma-inducible protein-10 (IP-10/CXCL10) is a chemoattractant for various leukocyte subsets and has been implicated in the pathogenesis of atherosclerosis. This study investigates whether IP-10 is expressed in human adipocytes and whether its release is regulated by body mass index (BMI) or immunological stimuli. METHODS: In cultures of human mature adipocytes and in vitro differentiated adipocytes, IP-10 expression under basal conditions and in the presence of IFNgamma, lipopolysaccharide (LPS) or interleukin (IL)-4 was characterized by reverse transcriptase-polymerase chain reaction, Luminex technology and immunofluorescence. RESULTS: IP-10 was expressed and secreted constitutively in most cultures of mature adipocytes from omental and subcutaneous (s.c.) depots. The association between IP-10 release and donor BMI was not significant. In in vitro differentiated adipocytes from s.c. and mammary depots and in mature s.c. adipocytes, IP-10 secretion was strongly upregulated by IFNgamma, whereas LPS or IL-4 did not affect IP-10 expression in s.c. mature adipocytes. Immunofluorescence confirmed IP-10 expression in adipocytes with abundant lipid droplets. CONCLUSION: Mature human adipocytes express and secrete the chemokine IP-10 and are thus identified as a novel cellular source of this disease-related immune mediator. IP-10 expression could be significantly induced by IFNgamma, but not by LPS, which points to both similar reactivities and functional differences between adipocytes and innate immune cells.

[Secretory activity of the adipocytes and comorbidities of obesity]. / Sekretorische Aktivität des Fettgewebes. Vom Energiespeicher zum endokrinen Organ.

Fatty tissue synthesizes and secretes a wide range of products that may be directly involved in the pathogenesis of the complications associated with obesity. These so-called adipokines may trigger or sustain a chronic inflammatory process. By manipulating the secretory function of fat cells, it might in future be possible to prevent the development of the metabolic and cardiovascular complications of obesity. Current data already suggest that weight reduction and certain substances with an anti-inflammatory action reduce the risk for the metabolic and cardiovascular complications of obesity. To date, however, the evidence available is only indirect, and is insufficient to definitively establish causal relationships between certain secretory products of adipocytes and the comorbidities of adiposity. Further clinical studies are needed.

Obesity and impaired fibrinolysis: role of adipose production of plasminogen activator inhibitor-1.

Obesity is the central promoter of the metabolic syndrome which also includes disturbed fibrinolysis in addition to hypertension, dyslipidaemia and impaired glucose tolerance/type 2 diabetes mellitus. Plasminogen activator inhibitor-1 (PAI-1) is the most important endogenous inhibitor of tissue plasminogen activator and uro-plasminogen activator, and is a main determinant of fibrinolytic activity. There is now compelling evidence that obesity and, in particular, an abdominal type of body fat distribution are associated with elevated PAI-1 antigen and activity levels. Recent studies established that PAI-1 is expressed in adipose tissue. The greater the fat cell size and the adipose tissue mass, the greater is the contribution of adipose production to circulating PAI-1. Experimental data show that visceral adipose tissue has a higher capacity to produce PAI-1 than subcutaneous adipose tissue. Studies in human adipocytes indicate that PAI-1 synthesis is upregulated by insulin, glucocorticoids, angiotensin II, some fatty acids and, most potently, by cytokines such as tumour necrosis factor-alpha and transforming growth factor-beta, whereas catecholamines reduce PAI-1 production. Interestingly, pharmacological agents such as thiazolidinediones, metformin and AT(1)-receptor antagonists were found to reduce adipose expression of PAI-1. In addition, weight loss by dietary restriction or comprehensive lifestyle modification is effective in lowering PAI-1 plasma levels. In conclusion, impaired fibrinolysis in obesity is probably also due to an increased expression of PAI-1 in adipose tissue. An altered function of the endocrine system and an impaired auto-/paracrine function at the fat cell levels may mediate this disturbance of the fibrinolytic system and thereby increase the risk for cardiovascular disease..

Effect of the angiotensin II receptor blocker candesartan on fibrinolysis in patients with mild hypertension.

AIM: Impaired fibrinolysis is frequently observed in patients with the metabolic syndrome. Aim of the study was to examine the short-term effect of angiotensin II receptor blockade on the fibrinolytic system. METHODS: Seventy-four patients with mild hypertension were randomly assigned to a 7-day treatment period with either 16 mg candesartan cilexetil or placebo. Several variables of the fibrinolytic system such as plasminogen activator inhibitor-1 (PAI-1) antigen and activity, tissue plasminogen activator (t-PA) antigen and activity as well as circulating t-PA/PAI-1 complexes were determined. RESULTS: At baseline, the body mass index but not blood pressure was positively associated with PAI-1 antigen (r=0.314, p<0.01) and PAI-1 activity (r=0.425, p<0.01) but negatively with t-PA activity (r=-0.187, p < 0.05). A 7-day treatment with 16 mg candesartan cilexetil resulted in a significant greater reduction of diastolic blood pressure (-10.3 +/- 10.8 mmHg vs.-5.8 +/- 8.5 mmHg, p=0.03). However, there was no significant effect of candesartan on all parameters of the fibrinolytic system under investigation, i.e. circulating PAI-1 antigen, PAI-1 activity, t-PA antigen, t-PA activity and t-PA/PAI-1 complexes. Furthermore, candesartan did not affect the characteristic circadian pattern of the variables of the fibrinolytic system. CONCLUSION: We conclude that short-term blockade of the angiotensin II receptor subtype 1 with candesartan does not have an impact on fibrinolysis in patients with mild hypertension.

HMG-CoA reductase inhibitor cerivastatin inhibits interleukin-6 expression and secretion in human adipocytes.

Human adipose tissue is a main contributor to plasma levels of pro-inflammatory cytokine IL-6. How IL-6 expression is regulated in adipocytes remains unclear. In the current study, we investigated the effect of the HMG-CoA reductase inhibitor, cerivastatin, on the production of IL-6 from cultured human adipocytes. Cerivastatin reduced both IL-6 mRNA and secretion in a dose- and time-dependent manner. The inhibitory effect on IL-6 mRNA was prevented by the intermediates of the cholesterol synthesis pathway, mevalonate and geranyl-geranyl-phyrophosphate (GGPP) but not by farnesyl-pyrophosphate. This suggests the involvement of geranylgeranyl-modified intermediates in the effect of cerivastatin on IL-6. Moreover, cerivastatin induced an inactivation of the phosphorylation of the p65 subunit of NFkappaB which was prevented by GGPP. Our data suggest that cerivastatin exerts an anti-inflammatory effect by down-regulating IL-6 levels in adipocytes, which seems to be mediated by reduced production of GGPP and interference with the NFkappaB pathway.

Effect of BMI and age on adipose tissue cellularity and differentiation capacity in women.

OBJECTIVE: To study the relation between body mass index (BMI) and age on the one hand and total number of human (pre-) adipocytes and preadipocyte differentiation capacity on the other hand. SUBJECTS: In total, 189 women undergoing surgical mammary reduction, age range 16-73 y, BMI range 19.7-39.7 kg/m(2). MEASUREMENTS: Differentiation of preadipocytes in primary culture was assessed by morphological criteria, and determination of glycerol-3-phosphate dehydrogenase after stimulation of the cells by standardized adipogenic conditions containing isobutyl-methylxanthine, troglitazone or both compounds. The total number of stromal cells (ie preadipocytes) and fat cells per gram of adipose tissue and per body as well as mature fat cell volume were calculated from isolated stromal cells and adipocytes, respectively, and anthropometric measures. RESULTS: BMI correlated positively to age, mature fat cell size and total number of adipocytes and stromal cells per body (r varying from 0.22 to 0.54, each P<0.05). In contrast, BMI correlated negatively to the number of adipocytes and stromal cells per gram of adipose tissue and the capacity of preadipocytes to differentiate (r varying from -0.20 to -0.37, each P<0.05). No significant correlation was observed between BMI and the ratio of stromal cells to adipocytes. The sample was also divided into three groups: BMI <25 kg/m(2) (lean), BMI 25-29.9 kg/m(2) (overweight) and BMI >/=30 kg/m(2) (obese). The overweight group showed a larger fat cell size but no increase in total fat cell or stromal cell number when compared to the lean subjects. The obese subjects showed larger stromal and fat cell numbers when compared to the lean subjects. Age did not independently correlate to the number of stromal cells or adipocytes per gram of adipose tissue or total body, nor with the capacity of preadipocytes to undergo differentiation and the ratio of stromal cells to adipocytes. CONCLUSION: There seems to be a constant ratio between the number of adipose tissue stromal cells and adipocytes independently of BMI and age in humans. During adipose tissue expansion, there seems to be both a continuous increase in fat cell size, and in stromal cell and adipocyte number, but the increase in fat cell size apparently precedes the increase in fat cell number. The differentiation capacity of the stromal cells appears to decrease with increasing BMI.

Relationship between IL-6, leptin and adiponectin and variables of fibrinolysis in overweight and obese hypertensive patients.

Impaired fibrinolysis is a common finding in obese humans. This condition is now considered as an established risk factor for thromboembolic complications. Furthermore, obesity is characterized by a specific pattern of circulating concentrations of fat-cell products interleukin-6 (IL-6), leptin, and adiponectin. The aim of our study was to investigate the relationship between these proteins and selected variables of the fibrinolytic system in 74 mildly hypertensive, overweight subjects. Circulating IL-6 and leptin levels showed a positive association with BMI (r = 0.24, p = 0.04 and r = 0.70, p < 0.0001), whereas adiponectin was not correlated to BMI. Interestingly, IL-6 was also positively associated with t-PA/PAI-1 complexes after adjustment for BMI and other anthropometric variables. Leptin was positively correlated with PAI-1 activity and antigen (r = 0.32, p = 0.006 and r = 0.37, p < 0.001, respectively) and negatively with t-PA activity (r = -0.27, p = 0.03). However, these associations lost significance after correction for BMI or HOMA, an insulin sensitivity index. In contrast, adiponectin levels were independently and negatively correlated with PAI-1 antigen (r = -0.26, p = 0.04, after correction for BMI). In conclusion, our study provides further evidence that IL-6, leptin, and adiponectin are associated with impaired fibrinolysis in overweight hypertensive humans.

Effects of leptin on the differentiation and metabolism of human adipocytes.

BACKGROUND: Leptin is an adipose protein regulating food intake in the hypothalamus. Animal studies have suggested that leptin also acts in an auto-/paracrine fashion on adipose cell function. OBJECTIVE: The aim of this study was to investigate the effects of leptin on the differentiation and metabolism of cultured human adipocytes. MATERIAL: Adipose tissue from young healthy, lean women (body mass index (BMI) <27 kg/m(2)) undergoing elective mammary reduction surgery and young obese individuals (BMI>40 kg/m(2)) undergoing laparoscopic gastric banding. METHODS: Human preadipocytes in primary culture were induced to undergo differentiation by defined adipogenic factors. Mature adipocytes were isolated by collagenase digestion and kept in culture suspension. Glycero-3-phosphate dehydrogenase (GPDH) activity was used as a marker of adipose differentiation; glucose uptake, lipolysis and PAI-1 secretion were measured as parameters of fat cell function. RESULTS: Human preadipocytes and adipocytes from lean and obese subjects expressed the long leptin receptor isoform and two of the three short forms as assessed by polymerase chain reaction (PCR). Leptin at a supraphysiological concentration induced a transient increase of GPDH activity, but had no effect on glucose uptake and PAI-1 secretion from human adipocytes. In addition, basal and isoproterenol-stimulated lipolysis as well as the antilipolytic action of insulin in human adipocytes was not significantly affected by leptin exposure. CONCLUSION: In contrast to animal data, the results of our experiments do not demonstrate significant effects of leptin on main metabolic functions of human adipocytes arguing against a local auto-/paracrine action of leptin in human adipose tissue.

Effect of angiotensin peptides on PAI-1 expression and production in human adipocytes.

Angiotensin (Ang) II is the active component of the renin-angiotensin-system (RAS), but its degradation products have also been shown to exhibit biological activity. This system, which mainly controls blood pressure and electrolyte homeostasis, was recently found to be completely expressed in human adipose tissue. The major determinant in the fibrinolytic system is the plasminogen activator inhibitor-1 (PAI-1). Both PAI-1 and components of the RAS are over-expressed in the obese state. We have recently shown that Ang II is able to induce PAI-1 expression and release via the AT1-receptor in human fat cells in primary culture, and have provided the first evidence that two metabolites, Ang III and Ang IV, may have a similar stimulatory effect on PAI-1 release. We have now performed additional experiments to further characterize the role of the angiotensin peptides in the production of PAI-1. Ang III and Ang IV showed a time- and dose-dependent stimulation of PAI-1 protein release. Concomitantly, mRNA-levels were markedly elevated. Using specific receptor blockers, all angiotensin peptides seem to induce PAI-1 expression via the angiotensin receptor subtype 1. However, components of the renin-angiotensin-system seem to play an important role in the control of fibrinolysis in adipose tissue. We conclude that PAI-1 production by adipose tissue may contribute to the elevated thromboembolic risk in obesity.

Angiotensin II and its metabolites stimulate PAI-1 protein release from human adipocytes in primary culture.

Plasminogen activator inhibitor (PAI)-1 is the main inhibitor of the fibrinolytic system and was recently shown to be produced by adipose cells. Obesity is associated with an increased production and release of PAI-1 protein. The aim of this study was to investigate the role of angiotensin (Ang) II and its degradation products for PAI-1 release from human adipose cells. For this purpose, we used the model of in vitro differentiated human adipocytes in primary culture. Exposure of human adipocytes to Ang II resulted in a dose- and time-dependent stimulation of PAI-1 release into the culture medium. The maximum effect of Ang II was found at a concentration of 10(-5) mol/L for 48 hours, increasing PAI-1 release by 276+/-53% compared with control cultures (P<0.05). This stimulation was preceded by an increase in specific PAI-1 mRNA copies by 65+/-12% (P<0.05), with a maximum after 6 hours. Incubation of adipocytes with 10(-5) mol/L Ang III and Ang IV, respectively, also resulted in a stimulation of PAI-1 release into the medium by 195+/-60% (P<0.05) and 142+/-24% (P<0.05), respectively, compared with control cultures. Addition of the angiotensin-receptor subtype 1 (AT(1)) blocker candesartan abolished the stimulatory action of Ang II and its metabolites, indicating that this effect is mediated by AT(1). Addition of the AT(1) blocker alone to unstimulated cultures reduced PAI-1 release by 41%+/-25% (P<0.05), suggesting that endogenous Ang II synthesis contributes to PAI-1 secretion from adipose tissue in an autocrine/paracrine manner. In conclusion, Ang II and its metabolites promote PAI-1 production and release by human fat cells and may contribute to the impairment of the fibrinolytic system typical for obesity. AT(1) receptor blockade reduces basal and abolishes Ang II-stimulated PAI-1 release from human adipocytes.

Beta-Adrenoceptor agonists and other cAMP elevating agents suppress PAI-1 production of human adipocytes in primary culture.

Recent studies showed that catecholamines contribute to the regulation of plasminogen activator inhibitor-1 (PAI-1) expression, at least in endothelial cells. Aim of this study was to examine the role of catecholamines on PAI-1 production by human adipocytes and, in particular, to clarify which adrenoceptor (AR) subtypes are involved. Addition of the unselective AR agonist isoproterenol led to a dose- and time-dependent suppression of PAI-1 mRNA and protein release in adipocytes from the subcutaneous and omental depot of obese subjects. A similar degree of suppression was observed in subcutaneous mammary adipocytes of lean women. This effect was mainly mediated via the beta2-adrenoceptor according to experiments using selective agonists. Moreover, addition of cAMP-elevating agents such as dibutyryl-cAMP, forskolin and the phosphodiesterase inhibitors isobutyl-methylxanthine and milrinone resulted in a reduction of PAI-1 of varying degrees. In conclusion, the results of this study support the assumption that catecholamines are able to down-regulate PAI-1 expression and secretion in human adipocytes via beta-adrenergic receptors.