Black locust (Robinia pseudoacacia L.) range contraction and expansion in Europe under changing climate.

Robinia pseudoacacia is one of the most frequent non-native species in Europe. It is a fast-growing tree of high economic and cultural importance. On the other hand, it is an invasive species, causing changes in soil chemistry and light regime, and consequently altering the plant communities. Previously published models developed for the potential distribution of R. pseudoacacia concerned 2070, and were based mainly on data from Western and Central Europe; here we extended these findings and included additional data from Eastern Europe. To fill the gap in current knowledge of R. pseudoacacia distribution and improve the reliability of forecasts, we aimed to (i) determine the extent to which the outcome of range modeling will be affected by complementing R. pseudoacacia occurrence data with sites from Central, Southeastern, and Eastern Europe, (ii) identify and quantify the changes in the availability of climate niches for 2050 and 2070, and discuss their impacts on forest management and nature conservation. We showed that the majority of the range changes expected in 2070 will occur as early as 2050. In comparison to previous studies, we demonstrated a greater eastward shift of potential niches of this species and a greater decline of potential niches in Southern Europe. Consequently, future climatic conditions will likely favor the occurrence of R. pseudoacacia in Central and Northeastern Europe where this species is still absent or relatively rare. There, controlling the spread of R. pseudoacacia will require monitoring sources of invasion in the landscape and reducing the occurrence of this species. The expected effects of climate change will likely be observed 20 years earlier than previously forecasted. Hence we highlighted the urgent need for acceleration of policies aimed at climate change mitigation in Europe. Also, our results showed the need for using more complete distribution data to analyze potential niche models.

Black locust (Robinia pseudoacacia) beloved and despised: a story of an invasive tree in Central Europe.

Robinia pseudoacacia, invaded many countries a long time ago and is now a common part of the Central European landscape. Positive economic but negative environmental impacts of Robinia result in conflicts of interest between nature conservation, forestry, urban landscaping, beekeepers and the public when defining management priorities. Because current legislation will determine the future distribution of Robinia in the landscape, a comprehensive view of this species is necessary. Although this species is well studied, most of the scientific papers deal with the economic aspects. Other information is published in local journals or reports. Therefore we reviewed the ecological and socio-economic impact of Robinia placing particular emphasis on the species' history, vegetation ecology, invasiveness and management. In Central Europe, Robinia is limited climatically by late spring frost combined with a short vegetation period, soil hypoxia, shade and frequent major disturbances. The long historical tradition of using Robinia for afforestation has resulted in its popularity as a widespread forest tree and it being an important part of the economy in some countries. The main reasons are its fast growth, valuable and resistant wood, suitability for amelioration, reclamation of disturbed sites and erosion control, honey-making and recently dendromass production. On the other hand, a side-effect of planting this nitrogen-fixing pioneer tree, very tolerant of the nature of the substrate, is its propagation and spread, which pose a problem for nature conservation. Robinia is considered invasive, threatening especially dry and semi-dry grasslands, some of the most species-rich and endangered types of habitat in the region, causing extinction of many endangered light-demanding plants and invertebrates due to changes in light regime, microclimate and soil conditions. Other often invaded habitats include open dry forests and shrubland, alluvial habitats, agrarian landscapes, urban and industrial environments and disturbed sites, e.g. post-fire sites, forest clearings or degraded forestry plantations. Without forestry, black locust abundance would decrease during succession in forests with highly competitive and shade tolerant trees and in mature forests it occurs only as admixture of climax trees. The limited pool of native woody species, lack of serious natural enemies and a dense cover of grasses and sedges can suppress forest succession and favour the development of Robinia monodominant stands over 70 years old. A stratified approach, which combines both tolerance in some areas and strict eradication at valuable sites, provides the best option for achieving a sustainable coexistence of Robinia with people and nature.

Forest herb species with similar European geographic ranges may respond differently to climate change.

Many phenological studies have shown that spring geophytes are very sensitive to climate change, responding by shifting flowering and fruiting dates. However, there is a gap in knowledge about climatic drivers of their distributions and range shifts under climate change. Here we aimed to estimate climate niche shifts for four widely distributed and common geophytes of the nemoral zone of Europe (Anemone nemorosa, Anemone ranunculoides, Convallaria majalis and Maianthemum bifolium) and to assess the threat level under various climate change scenarios. Using MaxEnt species distribution models and future climate change scenarios we found that the precipitation of the warmest quarter was the most important factor shaping their ranges. All species studied will experience more loss in the 2061-2080 period than in 2041-2060, and under more pessimistic scenarios. M. bifolium will experience the highest loss, followed by A. nemorosa, A. ranunculoides, and the smallest for C. majalis. A. ranunculoides will gain the most, while M. bifolium will have the smallest potential range expansion. Studied species may respond differently to climate change despite similar current distributions and climatic variables affecting their potential distribution. Even slight differences in climatic niches could reduce the overlap of future ranges compared to present. We expect that due to high dependence on the warmest quarter precipitation, summer droughts in the future may be particularly severe for species that prefer moist soils. The lack of adaptation to long-distance migration and limited availability of appropriate soils may limit their migration and lead to a decline in biodiversity and changes in European forests.

Catecholamine and insulin control of lipolysis in subcutaneous adipose tissue during long-term diet-induced weight loss in obese women.

The aim of this study was to investigate the evolution of the adrenergic and insulin-mediated regulation of lipolysis during different phases of a 6-mo dietary intervention. Eight obese women underwent a 6-mo dietary intervention consisting of a 1-mo very low-calorie diet (VLCD) followed by a 2-mo low-calorie diet (LCD) and 3-mo weight maintenance (WM) diet. At each phase of the dietary intervention, microdialysis of subcutaneous adipose tissue (SCAT) was performed at rest and during a 3-h hyperinsulinemic euglycemic clamp. Responses of dialysate glycerol concentration (DGC) were determined at baseline and during local perfusions with adrenaline or adrenaline and phentolamine before and during the last 30 min of the clamp. Dietary intervention induced a body weight reduction and an improved insulin sensitivity. DGC progressively decreased during the clamp, and this decrease was similar during the different phases of the diet. The adrenaline-induced increase in DGC was higher at VLCD and LCD compared with baseline condition and returned to prediet levels at WM. In the probe with adrenaline and phentolamine, the increase in DGC was higher than that in the adrenaline probe at baseline and WM, but it was not different at VLCD and LCD. The results suggest that the responsiveness of SCAT to adrenaline-stimulated lipolysis increases during the calorie-restricted phases due to a reduction of the α(2)-adrenoceptor-mediated antilipolytic action of adrenaline. At WM, adrenaline-stimulated lipolysis returned to the prediet levels. Furthermore, no direct relationship between insulin sensitivity and the diet-induced changes in the regulation of lipolysis was found.

Post-prandial endothelial dysfunction is ameliorated following weight loss in obese premenopausal women.

BACKGROUND: Endothelial dysfunction and postprandial hyperglycemia represent independent risk factors for cardiovascular diseases. Obesity is connected with endothelial impairments; however, it is unclear whether weight loss can modify endothelial function during the postprandial period. The aim of this study was to evaluate endothelial response (post-ischemic forearm blood flow, PIFBF) in a fasted state and following ingestion of 75 g glucose before and after very low caloric diet (VLCD). MATERIAL/METHODS: 40 obese premenopausal women (age 39.6 ± 7.8 years, BMI 34.3 ± 3.2 kg/m2) participated in 4-week very low caloric diet (VLCD, 800 kcal/day). Before and after VLCD, the baseline blood flow and PIFBF were measured using a mercury strain gauge plethysmography in fasting state as well as 1 hour after ingestion of 75 g glucose. RESULTS: Dietary intervention resulted in a 7% weight loss (p<0.05) and a decrease in insulin resistance index HOMA-IR (2.44 ± 1.25 vs. 1.66 ± 0.81, p<0.05). Before VLCD intervention, PIFBF following oral glucose challenge decreased by 8.2 ± 9.1 ml/min/100 g tissue, while after weight loss identical stimulus increased PIFBF by 4.2 ± 8.9 ml/min/100 g tissue (p<0.05). Plasma ICAM-1 and VCAM-1 decreased by 8% and 10%, respectively, throughout the study. CONCLUSIONS: Postprandial endothelial dysfunction is ameliorated following weight loss in obese women. This finding demonstrates the beneficial effects of weight reduction on atherosclerosis risk.

Robinia pseudoacacia-dominated vegetation types of Southern Europe: Species composition, history, distribution and management.

Knowledge of the species composition of invaded vegetation helps to evaluate an ecological impact of aliens and design an optimal management strategy. We link a new vegetation analysis of a large dataset to the invasion history, ecology and management of Robinia pseudoacacia stands across Southern Europe and provide a map illustrating Robinia distribution. Finally, we compare detected relationships with Central Europe. We show that regional differences in Robinia invasion, distribution, habitats and management are driven both by local natural conditions (climate and soil properties, low competitive ability with native trees) and socioeconomic factors (traditional land-use). Based on the classification of 467 phytosociological relevés we distinguished five broad vegetation types reflecting an oceanity-continentality gradient. The stands were heterogeneous and included 824 taxa, with only 5.8% occurring in more than 10% of samples, representing mainly hemerobic generalists of mesophilous, nutrient-rich and semi-shady habitats. The most common were dry ruderal stands invading human-made habitats. Among native communities, disturbed mesic and alluvial forests were often invaded throughout the area, while dry forests and scrub dominated in Balkan countries. Continuous, long-term and large-scale cultivation represent a crucial factor driving Robinia invasions in natural habitats. Its invasion should be mitigated by suitable management taking into account adjacent habitats and changing cultivation practices to select for native species. Robinia invasion has a comparable pattern in Central and Southern Europe, but there is a substantial difference in management and utilization causing heterogeneity of many South-European stands.

Effect of hyperinsulinemia and very-low-calorie diet on interstitial cytokine levels in subcutaneous adipose tissue of obese women.

Type 2 diabetes and obesity are associated with an enhanced release of a number of adipocytokines. Hyperinsulinemia, frequently present in type 2 diabetes and obesity, might be one of the drivers of the enhanced production of adipocytokines. The aim of this study was to investigate the interstitial levels of cytokines in subcutaneous adipose tissue (SCAT) in response to hyperinsulinemia and the effect of weight-reducing hypocaloric diet on this regulation in obese subjects. Thirteen obese premenopausal women participated in the study. Concentrations of seven cytokines were measured in plasma and in AT interstitial fluid collected by microdialysis during a euglycemic-hyperinsulinemic clamp and during control infusion of physiological saline. A subgroup of six women underwent a 4-wk very-low-calorie diet (VLCD). Microdialysis during the clamp was performed before and at the end of VLCD. Hyperinsulinemia induced an increase of monocyte chemoatractant protein (MCP-1) and IL-6 SCAT interstitial and plasma levels and elevated IL-8 levels in SCAT. The relative changes of IL-6 levels in the dialysate correlated with changes of IL-8 and MCP-1. The interstitial and plasma levels of IL-1ß, IL-10, TNFα, and plasminogen activator inhibitor (PAI-1) remained unchanged in response to hyperinsulinemia. VLCD resulted in enhancement of the hyperinsulinemia-induced augmentation of MCP-1, IL-6, and IL-8 interstitial levels. In conclusion, hyperinsulinemia upregulates the interstitial levels of MCP-1, IL-6, and IL-8 in SCAT in obese women, whereas it does not affect IL-1ß, IL-10, TNFα, and PAI-1 levels. Hypocaloric diet associated with weight reduction enhances the hyperinsulinemia-induced upregulation of MCP-1, IL-6, and IL-8 in SCAT.

Plasma levels and adipose tissue messenger ribonucleic acid expression of retinol-binding protein 4 are reduced during calorie restriction in obese subjects but are not related to diet-induced changes in insulin sensitivity.

CONTEXT: Retinol-binding protein 4 (RBP4) may play a role in the development of insulin resistance. OBJECTIVE: We investigated whether RBP4 adipose tissue mRNA expression and plasma level are related to insulin sensitivity during a diet-induced weight loss. DESIGN, SETTING, PATIENTS, AND INTERVENTION: Obese women followed a dietary intervention composed of a 4-wk very low-calorie diet (VLCD), a 2-month low-calorie diet, and 3-4 months of a weight maintenance (WM) phase. MAIN OUTCOME MEASURES: Clinical investigation was performed before and at the end of each phase. Insulin sensitivity was assessed with the euglycemic hyperinsulinemic clamp. Adipose tissue mRNA and plasma levels of RBP4 were determined using reverse transcription-quantitative PCR and ELISA, respectively. RESULTS: Weight and fat mass decreased during VLCD and were stabilized during WM. Glucose disposal rate increased during VLCD and remained elevated thereafter. Plasma levels of RBP4 decreased after VLCD and, although increasing at subsequent phases, remained lower than prediet values. Adipose tissue mRNA levels were diminished after VLCD, and increased during low-calorie diet and WM to reach basal values. Basal RBP4 levels or diet-induced variations of RBP4 were not different in lean women and two groups of obese women with high- and low-insulin sensitivity. CONCLUSIONS: Severe calorie restriction promotes a reduction in adipose tissue and plasma levels of RBP4. The study does not bring evidence for a role for RBP4 in the regulation of diet-induced changes in insulin sensitivity.

An increase in plasma adiponectin multimeric complexes follows hypocaloric diet-induced weight loss in obese and overweight pre-menopausal women.

Adiponectin is involved in the regulation of glucose and fatty acid metabolism, influences whole-body insulin sensitivity and protects arterial walls against the development of atherosclerosis. Plasma adiponectin is decreased in obese, insulin-resistant and Type 2 diabetic patients. Adiponectin circulates in plasma as high-, medium- and low-molecular-weight ('mass') forms (HMW, MMW and LMW respectively). The HMW form is believed to be closely associated with insulin sensitivity. The aim of the present study was to investigate whether diet-induced changes in body weight and insulin sensitivity were associated with changes in the quantity of adiponectin multimeric complexes. A total of 20 overweight or obese women (age, 39.4+/-9.5 years; body mass index, 32.2+/-6.4 kg/m(2)) underwent 12 weeks of low caloric diet (600 kcal/day less than energy requirements; where 1 kcal is approximately 4.184 kJ). Plasma samples were drawn before and after the study for biochemical analysis and Western blot detection of adiponectin multimeric complexes. The hypocaloric diet resulted in a weight reduction (89.8+/-16.4 kg compared with 83.1+/-15.6 kg; P<0.001) and an improvement in whole-body insulin sensitivity, as measured by HOMA (homoeostasis model assessment index; 1.9+/-0.8 compared with 1.5+/-0.7; P=0.013). Increases in the quantities of the HMW, MMW and LMW forms by 5.5, 8.5 and 18.1% respectively, were observed (P<0.05 for all of the forms). Total plasma adiponectin was increased by 36% with borderline significance (P=0.08). No correlations between changes in adiponectin complexes and changes in indices of insulin sensitivity were observed. In conclusion, diet-induced weight loss improved insulin sensitivity as well as increased the amount of HMW, MMW and LMW adiponectin complexes in plasma.

Timing Is Important: Unmanned Aircraft vs. Satellite Imagery in Plant Invasion Monitoring.

The rapid spread of invasive plants makes their management increasingly difficult. Remote sensing offers a means of fast and efficient monitoring, but still the optimal methodologies remain to be defined. The seasonal dynamics and spectral characteristics of the target invasive species are important factors, since, at certain time of the vegetation season (e.g., at flowering or senescing), plants are often more distinct (or more visible beneath the canopy). Our aim was to establish fast, repeatable and a cost-efficient, computer-assisted method applicable over larger areas, to reduce the costs of extensive field campaigns. To achieve this goal, we examined how the timing of monitoring affects the detection of noxious plant invaders in Central Europe, using two model herbaceous species with markedly different phenological, structural, and spectral characteristics. They are giant hogweed (Heracleum mantegazzianum), a species with very distinct flowering phase, and the less distinct knotweeds (Fallopia japonica, F. sachalinensis, and their hybrid F. × bohemica). The variety of data generated, such as imagery from purposely-designed, unmanned aircraft vehicle (UAV), and VHR satellite, and aerial color orthophotos enabled us to assess the effects of spectral, spatial, and temporal resolution (i.e., the target species' phenological state) for successful recognition. The demands for both spatial and spectral resolution depended largely on the target plant species. In the case that a species was sampled at the most distinct phenological phase, high accuracy was achieved even with lower spectral resolution of our low-cost UAV. This demonstrates that proper timing can to some extent compensate for the lower spectral resolution. The results of our study could serve as a basis for identifying priorities for management, targeted at localities with the greatest risk of invasive species' spread and, once eradicated, to monitor over time any return. The best mapping strategy should reflect morphological and structural features of the target plant and choose appropriate spatial, spectral, and temporal resolution. The UAV enables flexible data acquisition for required time periods at low cost and is, therefore, well-suited for targeted monitoring; while satellite imagery provides the best solution for larger areas. Nonetheless, users must be aware of their limits.

Black locust--successful invader of a wide range of soil conditions.

Black locust (Robinia pseudoacacia, BL), a species native to North America, has successfully invaded many types of habitats over the world. This study provides an overall assessment of BL soil conditions to determine the range of physical-chemical soil properties it can tolerate. 511 BL stands (for the soil types) and 33 permanent plots (for the soil chemistry) were studied in the Czech Republic. Relationships among different environmental variables (physical-chemical soil properties, vegetation characteristics and habitat conditions) were investigated and variables with the highest effect on species composition were detected. The results were compared with data in the literature for other parts of the secondary and native distributions of this species. This assessment showed that BL is able to tolerate extremely diverse soil physical-chemical conditions, from extremely acid to strongly alkaline, and from medium to highly base saturated soils with a gradient of different subsurface stoniness. Soil nitrate, N mineralization and nitrification rates also varied considerably and the concentrations of exchangeable phosphorus and ammonium were consistently low. N mineralization rate, incubated inorganic nitrogen and nitrates were positively correlated with base saturation and cation exchange capacity. The most common soil types were young soils (Cambisols, Leptosols, Arenosols, and coarsely textured Fluvisols). BL seems to be limited by water supply and soil aeration and prefers well aerated and drained soils, and tolerates desiccation but avoids compact soils and areas where the soils are frequently waterlogged. On steep slopes, BL was less vigorous, stunted and less competitive. By contrast, the tallest BL trees were found on sandy soils in a flat landscape. Number and share of nitrophytes in the herb layer were positively related to basic bedrock, soil reaction and N-NO3/N ratio. Soil reaction was determined as the most important environmental characteristic explaining the variability in BL species composition in the Czech Republic.

Global Invader Impact Network (GIIN): toward standardized evaluation of the ecological impacts of invasive plants.

Terrestrial invasive plants are a global problem and are becoming ubiquitous components of most ecosystems. They are implicated in altering disturbance regimes, reducing biodiversity, and changing ecosystem function, sometimes in profound and irreversible ways. However, the ecological impacts of most invasive plants have not been studied experimentally, and most research to date focuses on few types of impacts, which can vary greatly among studies. Thus, our knowledge of existing ecological impacts ascribed to invasive plants is surprisingly limited in both breadth and depth. Our aim was to propose a standard methodology for quantifying baseline ecological impact that, in theory, is scalable to any terrestrial plant invader (e.g., annual grasses to trees) and any invaded system (e.g., grassland to forest). The Global Invader Impact Network (GIIN) is a coordinated distributed experiment composed of an observational and manipulative methodology. The protocol consists of a series of plots located in (1) an invaded area; (2) an adjacent removal treatment within the invaded area; and (3) a spatially separate uninvaded area thought to be similar to pre-invasion conditions of the invaded area. A standardized and inexpensive suite of community, soil, and ecosystem metrics are collected allowing broad comparisons among measurements, populations, and species. The method allows for one-time comparisons and for long-term monitoring enabling one to derive information about change due to invasion over time. Invader removal plots will also allow for quantification of legacy effects and their return rates, which will be monitored for several years. GIIN uses a nested hierarchical scale approach encompassing multiple sites, regions, and continents. Currently, GIIN has network members in six countries, with new members encouraged. To date, study species include representatives of annual and perennial grasses; annual and perennial forbs; shrubs; and trees. The goal of the GIIN framework is to create a standard yet flexible platform for understanding the ecological impacts of invasive plants, allowing both individual and synthetic analyses across a range of taxa and ecosystems. If broadly adopted, this standard approach will offer unique insight into the ecological impacts of invasive plants at local, regional, and global scales.

Immunoassay for biochanin A.

Two variants of immunoassay for the determination of biochanin A (5,7-dihydroxy 4'-methoxy isoflavone), i.e., a radioimmunoassay (RIA) and an indirect ELISA, have been developed and evaluated. Both methods employ the same rabbit antiserum to a 7-O-carboxymethyl-5-hydroxy-4'-methoxyisoflavone-bovine serum albumin (BSA) conjugate. A 125I-labeled hapten-tyrosine methyl ester (TME) conjugate was used as a radioligand for the RIA. The indirect ELISA uses immunogen-coated microtitration plates and a peroxidase-labeled antirabbit Ig antibody. Both methods are specific for biochanin A with a comparable sensitivity (3.1 pg/tube for RIA; 5.3 pg/well for ELISA); however, their sensitivity to individual cross-reactants differs. The main cross-reactants are sissotrin (the cross-reactivity 15.7% for RIA; 120% for ELISA), 5-hydroxy, 4',7-dimethoxy isoflavone (51.5% for RIA; 46.5% for ELISA), prunetin (4.5% for RIA; 5.0% for ELISA), genistein (0.8% for RIA; 2.8% for ELISA) and formononetin (0.4% for RIA; 0.3% for ELISA). These methods were used for the analysis of biochanin A in alfalfa and in several nonleguminous plants.

Synthesis of hapten and conjugates of coumestrol and development of immunoassay.

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.

Weight loss improves the adipogenic capacity of human preadipocytes and modulates their secretory profile.

Calorie restriction-induced weight loss is accompanied by profound changes in adipose tissue characteristics. To determine the effect of weight loss on differentiation of preadipocytes and secretory capacity of in vitro differentiated adipocytes, we established cultures of these cells from paired subcutaneous adipose tissue biopsies obtained before and at the end of weight-reducing dietary intervention (DI) in 23 obese women. Based on lipid accumulation and the expression of differentiation markers, in vitro adipogenesis increased after weight loss and it was accompanied by enhanced expression of genes involved in de novo lipogenesis. This effect of weight loss was not driven by changes of peroxisome proliferator-activated receptor γ sensitivity to rosiglitazone. Weight loss also enhanced the expression of adiponectin and leptin while reducing that of monocyte chemoattractant protein 1 and interleukin-8 by cultured adipocytes. Thus, the weight-reducing (DI) increased adipogenic capacity of preadipocytes and shifted their secretion toward lower inflammatory profile. Reprogramming of preadipocytes could represent an adaptation to weight loss leading to partial restoration of preobese adipose tissue traits and thus contribute to the improvement of metabolic status. However, enhanced adipogenesis could also contribute to the unwanted weight regain after initial weight loss.

The Impact of Full-Length, Trimeric and Globular Adiponectin on Lipolysis in Subcutaneous and Visceral Adipocytes of Obese and Non-Obese Women.

Contribution of individual adiponectin isoforms to lipolysis regulation remains unknown. We investigated the impact of full-length, trimeric and globular adiponectin isoforms on spontaneous lipolysis in subcutaneous abdominal (SCAAT) and visceral adipose tissues (VAT) of obese and non-obese subjects. Furthermore, we explored the role of AMPK (5'-AMP-activated protein kinase) in adiponectin-dependent lipolysis regulation and expression of adiponectin receptors type 1 and 2 (AdipoR1 and AdipoR2) in SCAAT and VAT. Primary adipocytes isolated from SCAAT and VAT of obese and non-obese women were incubated with 20 µg/ml of: A) full-length adiponectin (physiological mixture of all adiponectin isoforms), B) trimeric adiponectin isoform or C) globular adiponectin isoform. Glycerol released into media was used as a marker of lipolysis. While full-length adiponectin inhibited lipolysis by 22% in non-obese SCAAT, globular isoform inhibited lipolysis by 27% in obese SCAAT. No effect of either isoform was detected in non-obese VAT, however trimeric isoform inhibited lipolysis by 21% in obese VAT (all p<0.05). Trimeric isoform induced Thr172 p-AMPK in differentiated preadipocytes from a non-obese donor, while globular isoform induced Ser79 p-ACC by 32% (p<0.05) and Ser565 p-HSL by 52% (p = 0.08) in differentiated preadipocytes from an obese donor. AdipoR2 expression was 17% and 37% higher than AdipoR1 in SCAAT of obese and non-obese groups and by 23% higher in VAT of obese subjects (all p<0.05). In conclusion, the anti-lipolytic effect of adiponectin isoforms is modified with obesity: while full-length adiponectin exerts anti-lipolytic action in non-obese SCAAT, globular and trimeric isoforms show anti-lipolytic activity in obese SCAAT and VAT, respectively.

Adipose tissue secretion and expression of adipocyte-produced and stromavascular fraction-produced adipokines vary during multiple phases of weight-reducing dietary intervention in obese women.

CONTEXT: Obesity is associated with altered plasma levels of adipokines involved in the development of insulin resistance and obesity-related metabolic disturbances. OBJECTIVE: The aim was to investigate diet-induced changes in adipokine production in sc abdominal adipose tissue (SAT) during a 6-month, multiphase, weight-reducing dietary intervention. DESIGN, SETTING, PARTICIPANTS, AND INTERVENTIONS: Forty-eight obese women followed a dietary intervention consisting of a very low-calorie diet (VLCD) (1 month), followed by a weight-stabilization (WS) period, which consisted of a low-calorie diet (2 months), and a weight-maintenance diet (3 months). MAIN OUTCOME MEASURES: Before and at the end of the VLCD and WS, samples of plasma and SAT were obtained. In a subgroup of 26 women, secretion of adipokines was determined in SAT explants, and in a subgroup of 22 women, SAT mRNA expression was measured. RESULTS: Body weight decreased and insulin sensitivity increased during the intervention. Plasma levels, SAT mRNA expression, and secretion rates of adipocyte-produced adipokines (leptin, serum amyloid A, and haptoglobin) decreased during the VLCD and increased during the WS period. Adipokines produced mainly from stroma-vascular cells (IL-6, IL-8, IL-10, IL-1Ra, TNFα, plasminogen activator inhibitor-1, and monocyte chemoattractant protein-1) increased or remained unchanged during VLCD and decreased to levels equal to or lower than prediet levels during the WS period. The diet-induced changes in homeostasis model assessment of insulin resistance correlated with changes in leptin plasma levels during VLCD, WS, and the entire dietary intervention period. CONCLUSIONS: Diet-induced regulation of adipokine production in SAT differs according to their cellular origin (adipocytes vs. stroma-vascular cells) and diet phase (VLCD vs. WS). Insulin-sensitivity changes were associated only with those of plasma leptin.

The impacts of road and walking trails upon adjacent vegetation: effects of road building materials on species composition in a nutrient poor environment.

Roads represent an important landscape element affecting both biotic and abiotic components. Alteration of soil properties along roads (addition of nutrients) is assumed to have a great impact on vegetation structure especially in nutrient poor ecosystems. Existing studies focus mainly on road dust. In our study we assessed the overall effects of roads upon adjacent alpine tundra vegetation and soils in Krkonose Mts, Czech Republic. Our aims were to (1) reconstruct the road-related changes using aerial data and GPS mapping to study colonization of roadside plant species; (2) assess the road effects on physical-chemical soil properties and vegetation composition along transects; and (3) propose conservation measures to stop further damage. Changes were reconstructed from historical multispectral aerial photography (1986 to 1997), measured by GPS device (1997, 2004), and accompanied by detailed soil (1998, 2000 and 2001) and vegetation (2000 and 2004) surveys along transects. Along alkaline roads, fast and profound shifts in physical-chemical soil properties (pH increased from 3.9 up to 7.6, base saturation from 9-30% up to 100%), and species composition were recorded. The roadside vegetation doubled in area during the studied decade. Stress-tolerant tundra species were replaced by meso- to nitrophilous species and species preferring man-made habitats. The intensity of changes depended significantly on the type of road material and the position relative to the road (slope position, distance from the road). Our findings support the assumption that alkaline gravel is the main cause of changes along roads in the area, and indicate the leading role of water transport in the soil and consequent vegetation alteration. To prevent the further damage we recommended replacement of alkaline gravel by granite, even though expensive and technically complicated. Based on our recommendations, the National Park authorities started to reconstruct the trails, although recovery is expected to be slow.

Worsening of obesity and metabolic status yields similar molecular adaptations in human subcutaneous and visceral adipose tissue: decreased metabolism and increased immune response.

CONTEXT: It is not known whether biological differences reported between sc adipose tissue (SAT) and visceral adipose tissue (VAT) depots underlie the pathogenicity of visceral fat. OBJECTIVE: We compared SAT and VAT gene expression according to obesity, visceral fat accumulation, insulin resistance, and presence of the metabolic syndrome. DESIGN: Subjects were assigned into four groups (lean, overweight, obese, and obese with metabolic syndrome). SETTING: Subjects were recruited at a university hospital. PATIENTS: Thirty-two women were included. MAIN OUTCOME MEASURES: Anthropometric measurements, euglycemic-hyperinsulinemic clamps, blood analyses, and computed tomography scans were performed, and paired samples of SAT and VAT were obtained for DNA microarray-based gene expression profiling. RESULTS: Considering the two fat depots together, 1125 genes were more and 1025 genes were less expressed in lean compared with metabolic syndrome subjects. Functional annotation clustering showed, from lean to metabolic syndrome subjects, progressive down-regulation of metabolic pathways including branched-chain amino acid, fatty acid, carbohydrate, and mitochondrial energy metabolism and up-regulation of immune response genes involved in toll-like receptor, TNF, nuclear factor-κB, and apoptosis pathways. Metabolism and immune response genes showed an opposite correlation with fat mass, fat distribution, or insulin resistance indices. These associations were similar in SAT and VAT, although about 1000 genes showed differential expression between SAT and VAT. CONCLUSIONS: The increase in adiposity and the worsening of metabolic status are associated with a coordinated down-regulation of metabolism-related and up-regulation of immune response-related gene expression. Molecular adaptations in SAT prove as discriminating as those in VAT.

Secretion of adiponectin multimeric complexes from adipose tissue explants is not modified by very low calorie diet.

OBJECTIVE: Adiponectin is a protein abundantly secreted by the adipose tissue (AT). Plasma adiponectin levels are decreased in obese, insulin-resistant, and type 2 diabetic patients. Various multimeric complexes, i.e. high-, middle-, and low-molecular weight isoforms (HMW, MMW and LMW), are present in plasma. Here, we investigated the effect of weight reducing diet on the distribution of adiponectin isoforms in plasma and on their secretion in AT explants from obese subjects. DESIGN: A total of 20 obese subjects (age 37.8+/-7.3 years, body mass index 33.9+/-5.0 kg/m(2)) underwent eight weeks of very low-calorie diet (VLCD). A needle biopsy of subcutaneous abdominal AT and blood samples were taken before and after dietary intervention. AT explants were incubated in culture medium for 4 h. ELISA assay and western blot analyses were used to identify adiponectin complexes in culture media and in plasma. RESULTS: The distribution of adiponectin polymers in plasma was different from that secreted in human AT explants. Before VLCD, the relative amount of HMW isoform was 75.5+/-9.1% of total adiponectin in culture media and 52.2+/-11.2% in plasma. Despite the diet-induced weight loss and improvement of insulin sensitivity, VLCD neither induced change in total adiponectin level nor in the ratio of HMW to total adiponectin in plasma and in culture media of AT explants. CONCLUSIONS: The profile of adiponectin polymeric isoforms secreted by AT explants into culture media differs from the plasma profile. A dietary intervention leading to weight loss and improvement of insulin sensitivity was not associated with modifications of AT secretion of total or HMW adiponectin.