Differences in plasminogen activator inhibitor 1 in subcutaneous versus omental adipose tissue in non-obese and obese subjects.

Human adipose tissue can produce plasminogen activator inhibitor-1 (PAI-1). It has been suggested that high levels of PAI-1 are of importance in enhanced cardiovascular disease observed among obese subjects, especially abdominally obese individuals. In the present study, we investigated the level of mRNA and production of PAI-1 in adipose tissue from two adipose tissue depots (omental vs. subcutaneous). Adipose tissue from both depots was obtained from obese (mean BMI, 46.9 kg/m 2) and non-obese (mean BMI, 23.9 kg/m 2) women. PAI-1 mRNA was measured both in fresh adipose tissue obtained immediately after surgery and after the adipose tissue (fragments) had been incubated for up to 72 h. In immediately frozen adipose tissue, PAI-1 mRNA expression was similar in omental and subcutaneous adipose tissue. No differences between obese and non-obese women were found. However, when adipose tissue fragments were cultured, PAI-1 mRNA and PAI-1 production were significantly higher in omental than in subcutaneous adipose tissue (p < 0.05). In the culture system, the production of PAI-1 in obese subjects was higher than in non-obese subjects in both subcutaneous (p < 0.05) and in omental adipose tissue (p = 0.19). In order to test whether these regional differences observed after incubation of the adipose tissue were due to differences in local accumulation of cytokines that may stimulate PAI-1 by a paracrine or autocrine manner, we investigated the expression of transforming growth factor beta1 (TGF-beta1) mRNA and tumor necrosis factor alpha (TNF-alpha) mRNA and protein. No differences between the two fat depots were found. In conclusion, no differences in PAI-1 expression between omental and subcutaneous adipose tissue were observed in biopsies frozen immediately after removal, but after incubation of adipose tissue (which somehow stimulates PAI-1 production), higher levels of PAI-1 were found in omental adipose tissue than in subcutaneous adipose tissue. Finally, PAI-1 production in adipose tissue from obese women was higher in non-obese women after incubation for 72 h.     

Potential for optimized production and use of rapeseed biodiesel. Based on a comprehensive real-time LCA case study in Denmark with multiple pathways

Purpose

Several factors contribute to the current increased focus on alternative fuels such as biodiesel, including an increasing awareness of the environmental impact of petrochemical (PC) oil products such as PC diesel, the continuously increasing price of PC oil, and the depletion of PC oil. For these reasons, the European Union has enacted a directive requiring each member state to ensure that the share of energy from renewable sources in transport be at least 10 % of the final consumption of energy by 2020 (The European Parliament and the Council 2009). This LCA study assesses the specific environmental impacts from the production and use of biodiesel as it is today (real-time), based on rapeseed oil and different types of alcohols, and using technologies that are currently available or will be available shortly. Different options are evaluated for the environmental improvement of production methods. The modeling of the LCA is based on a specific Danish biodiesel production facility.

Methods

The functional unit is “1,000 km transportation for a standard passenger car.” All relevant process stages are included, such as rapeseed production including carbon sequestration and N₂O balances, and transportation of products used in the life cycle of biodiesel. System expansion has been used to handle allocation issues.

Results and discussion

The climate change potential from the production and use of biodiesel today is 57 kg CO₂-eq/1,000 km, while PC diesel is 214 kg CO₂-eq/1,000 km. Options for improvement include the increased use of residual straw from rapeseed fields for combustion in a power plant where carbon sequestration is considered, and a change in transesterification from a conventional process to an enzymatic process when using bioethanol instead of PC methanol. This research also evaluates results for land use, respiratory inorganics potential, human toxicity (carc) potential, ecotoxicity (freshwater) potential, and aquatic eutrophication (N) potential. Different sources for uncertainty are evaluated, and the largest drivers for uncertainty are the assumptions embedded into the substitution effects. The results presented should not be interpreted as a blueprint for the increased production of biodiesel but rather as a benchmarking point for the present, actual impact in a well-to-wheels perspective of biodiesel, with options for improving production and use.

Conclusions

Based on this analysis, we recommend investigating additional options and incentives regarding the increased use of rape straw, particularly considering the carbon sequestration issues (from the perspective of potential climate change) of using bioalcohol instead of PC alcohol for the transesterification process.     

Pooled human platelet lysate versus fetal bovine serum—investigating the proliferation rate,chromosome stability and angiogenic potential of human adipose tissue-derived stem cells intended for clinical use

Background aimsBecause of an increasing focus on the use of adipose-derived stem cells (ASCs) in clinical trials, the culture conditions for these cells are being optimized. We compared the proliferation rates and chromosomal stability of ASCs that had been cultured in Dulbecco's modified Eagle's Medium (DMEM) supplemented with either pooled human platelet lysate (pHPL) or clinical-grade fetal bovine serum (FBS) (DMEM^pHPL versus DMEM^FBS).MethodsASCs from four healthy donors were cultured in either DMEM^pHPL or DMEM^FBS, and the population doubling time (PDT) was calculated. ASCs from two of the donors were expanded in DMEM^pHPL or DMEM^FBS and cultured for the final week before harvesting with or without the addition of vascular endothelial growth factor. We assessed the chromosomal stability (through the use of array comparative genomic hybridization), the expression of ASC and endothelial surface markers and the differentiation and angiogenic potential of these cells.ResultsThe ASCs that were cultured in pHPL exhibited a significantly shorter PDT of 29.6 h (95% confidence interval, 22.3–41.9 h) compared with those cultured in FBS, for which the PDT was 123.9 h (95% confidence interval, 95.6–176.2 h). Comparative genomic hybridization analyses revealed no chromosomal aberrations. Cell differentiation, capillary structure formation and cell-surface marker expression were generally unaffected by the type of medium supplement that was used or by the addition of vascular endothelial growth factor.ConclusionsWe observed that the use of pHPL as a growth supplement for ASCs facilitated a significantly higher proliferation rate compared with FBS without compromising genomic stability or differentiation capacity.     

The chromophore structure of the long-lived intermediate of the C128T channelrhodopsin-2 variant

The photocycle of the light-activated channel, channelrhodopsin-2 C128T, has been studied by resonance Raman (RR) spectroscopy focussing on the intermediates P380 and P353 that constitute a side pathway in the recovery of the parent state. The P353 species displays a UV–vis absorption spectrum with a fine-structure reminiscent of the reduced-retro form of bacteriorhodopsin, whereas the respective RR spectra differ substantially. Instead, the RR spectra of the P380/P353 intermediate couple are closely related to that of a free retinal in the all-trans configuration. These findings imply that the parent state recovery via P380/P353 involves the transient hydrolysis and re-formation of the retinal–protein linkage.     

Strong microsite control of seedling recruitment in tundra

The inclusion of environmental variation in studies of recruitment is a prerequisite for realistic predictions of the responses of vegetation to a changing environment. We investigated how seedling recruitment is affected by seed availability and microsite quality along a steep environmental gradient in dry tundra. A survey of natural seed rain and seedling density in vegetation was combined with observations of the establishment of 14 species after sowing into intact or disturbed vegetation. Although seed rain density was closely correlated with natural seedling establishment, the experimental seed addition showed that the microsite environment was even more important. For all species, seedling emergence peaked at the productive end of the gradient, irrespective of the adult niches realized. Disturbance promoted recruitment at all positions along the environmental gradient, not just at high productivity. Early seedling emergence constituted the main temporal bottleneck in recruitment for all species. Surprisingly, winter mortality was highest at what appeared to be the most benign end of the gradient. The results highlight that seedling recruitment patterns are largely determined by the earliest stages in seedling emergence, which again are closely linked to microsite quality. A fuller understanding of microsite effects on recruitment with implications for plant community assembly and vegetation change is provided.     

Seedling stage strategies as a means of habitat specialization in herbaceous plants

The regeneration niche has been little investigated in studies of community assembly and plant distribution. We examined adaptive associations between seedling traits and habitat specialization. Two habitat contrasts were investigated across several evolutionary lineages of angiosperms: species specialized to forest vs. open habitats and to dry vs. wet habitats. We also tested whether effects of shade and drought vary independently or, alternatively, if shade may amplify effects on drought-stressed plants. Seedling response in terms of growth rate, height, slenderness, specific leaf area (SLA) and degree of elongation (longest internode; petiole or leaf-sheath depending on species' morphology) to light and watering treatments was assessed. We used a factorial design involving three light regimes and two watering frequencies. The open-shaded habitat contrast and the dry-wet habitat contrast were investigated using six and five pairs of congeneric species, respectively. The congeneric species pair design controlled for confounding effects of evolutionary history prior to divergence in habitat specialization. Seedling growth rate generally decreased with shade and reduced watering frequency. Plant height was generally largest at intermediate light. Specialization to shaded habitats was associated with a more conservative growth strategy, i.e. showing a more modest growth response to increasing light. Species from all habitats showed the highest relative elongation at intermediate light, except for the moist-habitat species, for which elongation increased with shade. Contrary to our expectations, species from dry habitats grew bigger than species from moist habitats in all treatments. SLA responded to the light treatment, but not to watering regime. The contrasting light and moisture conditions across habitats appear to not have selected for differences in SLA. We conclude that seedling phase strategies of resource allocation in temperate herbs contribute to their habitat specialization. Habitat-specific seedling strategies and trade-offs in response to resource availability and environmental conditions may be important to adaptive specialization.     

Inducing Dendritic Growth in Cultured Sympathetic Neurons

The shape of the dendritic arbor determines the total synaptic input a neuron can receive ^1-3, and influences the types and distribution of these inputs ^4-6. Altered patterns of dendritic growth and plasticity are associated with impaired neurobehavioral function in experimental models ⁷, and are thought to contribute to clinical symptoms observed in both neurodevelopmental disorders ^8-10 and neurodegenerative diseases ^11-13. Such observations underscore the functional importance of precisely regulating dendritic morphology, and suggest that identifying mechanisms that control dendritic growth will not only advance understanding of how neuronal connectivity is regulated during normal development, but may also provide insight on novel therapeutic strategies for diverse neurological diseases.Mechanistic studies of dendritic growth would be greatly facilitated by the availability of a model system that allows neurons to be experimentally switched from a state in which they do not extend dendrites to one in which they elaborate a dendritic arbor comparable to that of their in vivo counterparts. Primary cultures of sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rodents provide such a model. When cultured in defined medium in the absence of serum and ganglionic glial cells, sympathetic neurons extend a single process which is axonal, and this unipolar state persists for weeks to months in culture ^14,15. However, the addition of either bone morphogenetic protein-7 (BMP-7) ^16,17 or Matrigel ¹⁸ to the culture medium triggers these neurons to extend multiple processes that meet the morphologic, biochemical and functional criteria for dendrites. Sympathetic neurons dissociated from the SCG of perinatal rodents and grown under defined conditions are a homogenous population of neurons ¹⁹ that respond uniformly to the dendrite-promoting activity of Matrigel, BMP-7 and other BMPs of the decapentaplegic (dpp) and 60A subfamilies ^17,18,20,21. Importantly, Matrigel- and BMP-induced dendrite formation occurs in the absence of changes in cell survival or axonal growth ^17,18.Here, we describe how to set up dissociated cultures of sympathetic neurons derived from the SCG of perinatal rats so that they are responsive to the selective dendrite-promoting activity of Matrigel or BMPs.     

Colocalization of (3H)-adenosine accumulation and GABA immunoreactivity in the chicken and rabbit retinas

Using combined autoradiography and immunohistochemistry, we have compared (3H)-adenosine accumulation and GABA immunoreactivity in the chicken and rabbit retinas. Colocalization of the two markers was observed in a subset of amacrine cells and in certain cell bodies in the ganglion cell layer in both species and in a few horizontal cells in the chicken retina. Cells that contained only (3H)-adenosine or GABA were also seen. The degree of colocalization differed greatly between the two species. The results demonstrate a morphological relationship between the adenosine and GABA systems and provides information on the possible anatomical substrates underlying at least some types of functional interactions.     

Isolation of serum amyloid A protein by small-scale hydrophobic interaction chromatography and two-dimensional electrophoresis

A recently introduced technique to isolate serum amyloid A protein is hydrophobic interaction chromatography combined with two-dimensional electrophoresis with immobilized pH gradients. A modification of the original version of this technique is presented. Mouse serum was subjected to hydrophobic interaction chromatography on a small scale, and the eluate was applied directly to two-dimensional electrophoresis. Simple electropherogramss with optimal resolution of serum amyloid A protein were obtained. The presented technique facilitates isolation of serum amyloid A protein from small blood volumes, and might also be adapted to alternative applications.     

Enhancement of local species richness in tundra by seed dispersal through guts of muskox and barnacle goose

The potential contribution of vertebrate-mediated seed rain to the maintenance of plant community richness in a High Arctic ecosystem was investigated. We analyzed viable seed content in dung of the four numerically most important terrestrial vertebrates in Northeast Greenland – muskox (Ovibos moschatus), barnacle goose (Branta leucopsis), Arctic fox (Alopex lagopus) and Arctic hare (Lepus arcticus). High numbers of plant propagules were found in the dung of muskox and barnacle goose. Seeds of many plant species were found in the faeces of one vertebrate species only. Propagule composition in barnacle goose droppings was relatively uniform over samples, with a high abundance of the nutritious bulbils of Polygonum viviparum (Bistorta vivipara), suggesting that geese have a narrow habitat preference and feed selectively. Propagule composition in muskox dung was diverse and heterogeneous among samples, suggesting a generalist approach in terms of food selection and the haphazard ingestion of plant propagules with foliage. The species composition of plant propagules in dung samples was different from that of the receiving plant communities (in terms of the Sørensen and Czekanowski dissimilarity indices), and dung deposition, especially by muskox, often brought new species to the receiving community. The results suggest that endozoochorous propagule dispersal in the Arctic has a great potential in the generation and maintenance of local species richness, albeit being little specialized. It is further suggested that endozoochory is an important means of long-distance dispersal and, thereby, of plant migration in response to climate change.