Neuropeptide Y in the noradrenergic neurones induces obesity and inhibits sympathetic tone in mice.

AIM: Neuropeptide Y (NPY) co-localized with noradrenaline in central and sympathetic nervous systems seems to play a role in the control of energy metabolism. In this study, the aim was to elucidate the effects and pathophysiological mechanisms of increased NPY in catecholaminergic neurones on accumulation of body adiposity. METHODS: Transgenic mice overexpressing NPY under the dopamine-beta-hydroxylase promoter (OE-NPY(DßH) ) and wild-type control mice were followed for body weight gain and body fat content. Food intake, energy expenditure, physical activity, body temperature, serum lipid content and markers of glucose homoeostasis were monitored. Thermogenic and lipolytic responses in adipose tissues, and urine catecholamine and tissue catecholamine synthesizing enzyme levels were analysed as indices of sympathetic tone. RESULTS: Homozygous OE-NPY(DßH) mice showed significant obesity accompanied with impaired glucose tolerance and insulin resistance. Increased adiposity was explained by neither increased food intake or fat absorption nor by decreased total energy expenditure or physical activity. Adipocyte hypertrophy and decreased circulating lipid levels suggested decreased lipolysis and increased lipid uptake. Brown adipose tissue thermogenic capacity was decreased and brown adipocytes filled with lipids. Enhanced response to adrenergic stimuli, downregulation of catecholamine synthesizing enzyme expressions in the brainstem and lower adrenaline excretion supported the notion of low basal catecholaminergic activity. CONCLUSION: Increased NPY in catecholaminergic neurones induces obesity that seems to be a result of preferential fat storage. These results support the role of NPY as a direct effector in peripheral tissues and an inhibitor of sympathetic activity in the pathogenesis of obesity.

Resorption rate assessment of adipose tissue-engineered constructs by intravital magnetic resonance imaging.

Engineering of adipose tissue by implantation of preadipocytes within biodegradable materials has already been extensively reported. However, a method that allows to accurately determine the resorption rate of adipose tissue constructs has not been described to date. The purpose of this study was to determine whether the non-invasive and non-destructive technique of magnetic resonance imaging (MRI) could be used to assess the resorption rate of adipose tissue substitutes after injection of human preadipocytes within fibrin into athymic nude mice. Different concentrations of undifferentiated preadipocytes were injected within fibrin into athymic nude mice. Two days, 3 months and 6 months post-implantation, the mice were anaesthetised and an MRI was performed using a 9.4 Tesla device in order to determine both volume and resorption rate of the implants. Subsequently, the specimens were explanted and qualitative analysis of adipose tissue formation was performed by histological examination. After implantation, a progressive resorption of all constructs was macroscopically observed. Implants could be easily visualised and delimited from the surrounding tissues by MRI. Magnetic resonance analysis demonstrated a resorption rate of the implants of 99-100% at 6 months, which was also confirmed by histological analysis. In the remaining implants, formation of human adipose tissue could be immunohistologically confirmed. Here, we show that MRI provides an efficient and non-invasive method for the assessment of implant resorption in adipose tissue engineering.

Biocompatible photocrosslinked poly(ester anhydride) based on functionalized poly(epsilon-caprolactone) prepolymer shows surface erosion controlled drug release in vitro and in vivo.

Star-shaped poly(epsilon-caprolactone) oligomers functionalized with succinic anhydride were used as prepolymers to prepare photocrosslinked poly(ester anhydride) to evaluate their in vivo drug delivery functionality and biocompatibility. Thus, in this work, erosion, drug release and safety of the photocrosslinked poly(ester anhydride) were examined in vitro and in vivo. A small water-soluble drug, propranolol HCl (M(w) 296 g/mol, solubility 50 mg/ml), was used as the model drug in an evaluation of the erosion controlled release. Drug-free and drug-loaded (10-60% w/w) poly(ester anhydride) discoids eroded in vitro (pH 7.4 buffer, +37 degrees C) linearly within 24-48 h. A strong correlation between the polymer erosion and the linear drug release in vitro was observed, indicating that the release had been controlled by the erosion of the polymer. Similarly, in vivo studies (s.c. implantation of discoids in rats) indicated that surface erosion controlled drug release from the discoids (drug loading 40% w/w). Oligomers did not decrease cell viability in vitro and the implanted discoids (s.c., rats) did not evoke any cytokine activity in vivo. In summary, surface erosion controlled drug release and the safety of photocrosslinked poly(ester anhydride) were demonstrated in this study.

Comparison of pre-adipocyte yield, growth and differentiation characteristics from excised versus aspirated adipose tissue.

Adipose tissue precursor cells (pre-adipocytes) are part of a stromal vascular fraction that can be easily isolated from fat tissue. Adipose tissue can be harvested by 2 methods: aspiration and excision. We analyzed whether the pre-adipocyte yield, growth characteristics and ability to differentiate into mature adipose tissue are influenced by the type of harvesting procedure. Adipose tissue was simultaneously harvested from the abdomen by surgical excision or aspiration according to the Coleman procedure in 10 individuals. This permitted inter- and intra-individual comparisons. Cell viability and yield were determined directly after isolation of pre-adipocytes. The growth kinetics were investigated in culture. Furthermore, pre-adipocytes were cultured under adipogenic conditions to compare their differentiation potential. The number of viable pre-adipocytes was significantly higher after excision of adipose tissue compared to aspiration. The proliferation kinetic was not influenced by the type of harvesting. No differences were observed in the differentiation potential of the pre-adipocytes between both groups. Compared to excision, aspiration of adipose tissue negatively affects the yield of pre-adipocytes. However, growth characteristics and differentiation potential of viable cultured cells are not influenced by the type of surgical harvesting. Due to its reduced donor site morbidity, we conclude that aspiration of adipose tissue is a valid harvesting method for isolation of pre-adipocytes.

Serum adiponectin and resistin levels in major depressive disorder.

OBJECTIVE: To examine the role of the adipose-tissue-derived low-grade inflammation markers adiponectin and resistin in major depressive disorder (MDD) in a population-based sample. METHOD: Serum levels of adiponectin and resistin were measured from 70 DSM-IV MDD subjects and 70 healthy controls. Depression severity was assessed with the 29-item Hamilton Depression Rating Scale. RESULTS: The MDD group had lowered serum adiponectin levels. Regression modelling with adjustments for age, gender, overweight, several socioeconomic and lifestyle factors, coronary heart disease and metabolic syndrome showed that each 5.0 microg/ml decrease in serum adiponectin increased the likelihood of MDD by approximately 20% (P = 0.01). The resistin levels correlated with atypical (P = 0.02), but not with typical depressive symptoms (P = 0.12). CONCLUSION: Our findings suggest that the lowered adiponectin levels in MDD are depression-specific and not explained by conventional low adiponectin-related factors such as such as coronary heart disease and metabolic disorders.

In vivo delivery of a peptide, ghrelin antagonist, with mesoporous silicon microparticles.

Peptides may represent potential treatment options for many severe illnesses. However, they need an effective delivery system to overcome rapid degradation after their administration. One possible way to prolong peptide action is to use particulate drug delivery systems. In the present study, thermally hydrocarbonized mesoporous silicon (THCPSi) microparticles (38-53 microm) were studied as a peptide delivery system in vivo. D-lys-GHRP6 (ghrelin antagonist, GhA) was used as a model peptide. The effects of GhA-loaded THCPSi microparticles on food intake (s.c., GhA dose 14 mg/kg) and on blood pressure (s.c., GhA dose 4 mg/kg) were examined in mice and rats, respectively. In addition, the effects of THCPSi microparticles (2 mg) on cytokine secretion in mice after single s.c. administration were examined by determining several cytokine plasma concentrations. The present results demonstrate that GhA can be loaded into THCPSi microparticles with a high loading degree (20% w/w). GhA loaded THCPSi microparticles inhibited food intake for a prolonged time, and increased blood pressure more slowly than encountered with a GhA solution. Furthermore, THCPSi microparticles did not increase cytokine activity. The present results suggest that THCPSi might be used as a drug delivery system for peptides.

Effect of protein, fat, carbohydrate and fibre on gastrointestinal peptide release in humans.

Short-term regulation of food intake controls what, when and how much we eat within a single day or a meal. This regulation results from an integrated response to neural and humoral signals that originate from the brain, gastrointestinal (GI) tract and adipose tissue. In the GI tract, multiple sites including the stomach, duodenum, distal small intestine, colon, and pancreas are involved in this process. Ingested food evokes satiety by mechanical stimulation and by release of peptides in the GI tract. The intestine in particular plays a key role in satiety through various peptides secreted in response to food. Many of the intestinal peptides inhibit also gastric emptying thus enhancing gastric mechanoreceptor stimulation. In this review, the current knowledge about the effects of different macronutrients and fibre on the release of GI satiety-related peptides in humans is discussed.

Effect of medium- and long-chain fatty acid diets on PPAR and SREBP-1 expression and glucose homeostasis in ACBP-overexpressing transgenic rats.

AIM: Acyl-CoAs are important intermediates and regulators of lipid metabolism. Binding proteins like acyl-CoA binding protein (ACBP) can influence their regulatory functions. ACBP has also been shown to exert direct effects on gene regulation in vitro. As the physiological relevance of ACBP in the regulation of lipid metabolism under high fat diets is unclear, we investigated the influence of such diets on the metabolic responses in ACBP-overexpressing rats. METHODS: A transgenic rat line overexpressing the ACBP gene was used to study the effects of 4 weeks of feeding with medium- (MC) or long-chain (LC) fatty acid-containing diets. Glucose tolerance tests were performed. Expression of transcription factors was measured by quantitative RT-PCR and protein levels of AMP-activated protein kinase were determined by western blotting. RESULTS: Transgenic animals fed the MC diet had an improved glucose tolerance and lower serum insulin levels compared with controls. Their liver PPARgamma (by 43%) and SREBP-1 (by 35%) mRNA levels were found to be decreased, while adipose tissue PPARgamma expression was increased by 31%. Tg animals fed the LC diet did not exhibit changes in glucose or insulin levels but exhibited increased mRNA levels of liver PPARs and SREBP-1 (1.5-3.5 times) and decreased protein levels of AMPKalpha (by 48%). CONCLUSIONS: Our results demonstrate that ACBP overexpression affects metabolic responses to diets with distinct difference in their fatty acid chain lengths. The molecular regulatory mechanism behind these effects seems to be an ACBP-induced tissue-specific regulation of expression of PPARs and SREBP.

Evoked EEG patterns during burst suppression with propofol.

BACKGROUND: During EEG suppression with isoflurane or sevoflurane anaesthesia, median nerve stimulation causes cortical responses of two kinds: an N20 wave with a latency of 20 ms and an EEG burst with a latency of 200 ms. We tested the possibility that median nerve stimulation during EEG suppression with propofol would cause an EEG response that was consistent enough to be of use for neuromonitoring. METHODS: Eight patients were anaesthetized with propofol to allow burst suppression. Electrical stimulation of the median nerve was applied during general anaesthesia and the EEG was measured. RESULTS: The EEG response to a painful stimulus had four successive components: (i) N20 and P22 potentials, reflecting activation of fast somatosensory pathways; (ii) a high-amplitude negative wave, possibly reflecting activation of the somatosensory cortex SII bilaterally; (iii) a burst (i.e. a negative slow wave with superimposed 10 Hz activity, probably reflecting an arousal mechanism); and (iv) a 13-15 Hz spindle, probably originating from the thalamus, similar to sleep spindles. These could be seen separately and in different combinations. Bursts and spindles during burst suppression were also seen without stimulation. In deepening propofol anaesthesia, spindles were seen in the continuous EEG before burst suppression was achieved. In deep anaesthesia, spindles were seen when bursts had ceased, and painful stimuli evoked sharp waves without subsequent bursts. CONCLUSION: In addition to SSEP (somatosensory evoked potentials), three different evoked responses are noted that could be useful for clinical monitoring.

Tibial nerve somatosensory evoked potentials during EEG suppression in sevoflurane anaesthesia.

OBJECTIVES AND METHODS: Cortical tibial nerve somatosensory evoked potentials (TSEPs) were recorded from 10 subjects in sevoflurane anaesthesia in order to study TSEP during EEG suppression. RESULTS: With a stimulation frequency less than one per second the major component was a positive wave which had maximal amplitude parietally ipsilaterally to stimulus and mean latency of 46.1 ms. It probably corresponds to the P40 wave. It was preceded by a widespread smaller positive wave, which corresponds to the subcortical P30 wave. In two patients a high amplitude negative wave, a couple of milliseconds before the positive wave, and maximal parietally contralateral to stimulus, was seen. All later waves were absent. CONCLUSION: The results are in agreement with our previous results from median nerve SEPs showing that the first cortical response from primary somatosensory cortex is enhanced, and later waves are suppressed. Hence, recording TSEPs during EEG suppression provides a way to record the activity of the primary somatosensory cortex accurately and rapidly due to the very good signal to noise ratio, so that even single responses to stimuli can be seen without averaging. Our results suggest that new cortical generators, which are not recordable awake, may be discovered in some patients.