Current agri-environmental policies dismiss varied perceptions and discourses on management of traditional rural biotopes

Traditional rural biotopes (TRBs) are threatened habitats that host significant biodiversity and several ecosystem services, and depend on active management such as low-intensity grazing. The current study explores private landowners’ decision-making on TRB management and abandonment within a social-ecological system framework. We provide insight into supporting resilience of TRB systems in the face of agricultural modernization. Using a mixed methods approach with content analysis and Q analysis, we demonstrate that TRB management fosters cultural, biological, aesthetic, and utilitarian values. These are reflected in different ways through conservationist’s, profit-oriented farmer’s, landscape manager’s, and landscape admirer’s discourses on TRB management. Overall, management reinforces landowners’ place attachment, and reflects an approach to landscapes as spatial representations of cultural heritage and identity over multiple generations. Landowners consider TRB pasturage and its social-ecological outcomes motivating and rewarding. Giving up grazing cattle and perceived bureaucracy of national agri-environment scheme contribute to TRB abandonment. Landowners point out that current policies detach TRB management from what is seen as “regular agriculture”, and the focus on monetary compensation bypasses the multiple values tied to TRB management. Based on our results, we suggest that promoting TRBs requires reconfiguring the current arrangement of remedial management payments and adopting a more participatory governance approach. Locally, resilience of TRB systems relies on the connections between landowners and landscapes that foster sense of place and landscape identity, which can be supported by knowledge sharing and collaborative grazing efforts among landowners.peerReviewe

Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation-7

Sp, dashed area), secreted proteins without signal peptide (ex, white area) and intracellular proteins (in, dark area). The percentage of the proteins present in each group is reported in the scheme. One hundred per cent is referred to a total number of 73 identified proteins. . Venn diagram of proteins expressed under the different culture conditions; n represents protein number in each condition.<p><b>Copyright information:</b></p><p>Taken from "Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation"</p><p>http://www.biomedcentral.com/1471-2199/9/26</p><p>BMC Molecular Biology 2008;9():26-26.</p><p>Published online 26 Feb 2008</p><p>PMCID:PMC2279142.</p><p></p

Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation-1

Sp, dashed area), secreted proteins without signal peptide (ex, white area) and intracellular proteins (in, dark area). The percentage of the proteins present in each group is reported in the scheme. One hundred per cent is referred to a total number of 73 identified proteins. . Venn diagram of proteins expressed under the different culture conditions; n represents protein number in each condition.<p><b>Copyright information:</b></p><p>Taken from "Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation"</p><p>http://www.biomedcentral.com/1471-2199/9/26</p><p>BMC Molecular Biology 2008;9():26-26.</p><p>Published online 26 Feb 2008</p><p>PMCID:PMC2279142.</p><p></p

Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation-0

At day 3 and day 14 of differentiation as compared to day 0. The data are representative of three independent experiments. Microphotographs of hMADS cells differentiated into adipocytes and osteoblasts at day 14. Bar scale = 50 μm. Representative gel of secreted proteins from hMADS cells at day 0 and day 3 of differentiating adipocytes (adipo) and osteoblasts (osteo) after 6 h of incubation. The gel is representative of 3 independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation"</p><p>http://www.biomedcentral.com/1471-2199/9/26</p><p>BMC Molecular Biology 2008;9():26-26.</p><p>Published online 26 Feb 2008</p><p>PMCID:PMC2279142.</p><p></p

Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation-2

Ented in the pie chart. For each cluster the percentage of proteins included is reported.<p><b>Copyright information:</b></p><p>Taken from "Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation"</p><p>http://www.biomedcentral.com/1471-2199/9/26</p><p>BMC Molecular Biology 2008;9():26-26.</p><p>Published online 26 Feb 2008</p><p>PMCID:PMC2279142.</p><p></p

Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation-3

(), BIGH3 (), PTX3 () and PAI-1 () has been analyzed by Western blot. The bar graphs report the levels of expression of every single candidate as the mean of three independent experiments after 6 h of incubation. The values are indicated as arbitrary units. *: p < 0.05. Two μg of secreted proteins have been loaded for each gel.<p><b>Copyright information:</b></p><p>Taken from "Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation"</p><p>http://www.biomedcentral.com/1471-2199/9/26</p><p>BMC Molecular Biology 2008;9():26-26.</p><p>Published online 26 Feb 2008</p><p>PMCID:PMC2279142.</p><p></p

Adipocyte differentiation of hMADS cells requires the activation of both Epac- and PKA-dependent signaling pathways.

<p>Two-day post-confluent hMADS cells were induced to differentiate with 1 µM Dex, 0.86 µM insulin (DI), in the presence of 200 µM of the Epac-selective cAMP analog 8-pCPT-2′-O-Me-cAMP (007) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#pone.0034114-Christensen1" target="_blank">[37]</a>, 100 µM of PKA selective cAMP analog 6-MB-cAMP (MB) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#pone.0034114-Christensen1" target="_blank">[37]</a>, either separately or in combination (007+MB) from day 0 to day 3. Rosi (0.5 µM) was present from day 3–9. Undifferentiated hMADS cells were taken as control. (A) The panel shows cells on day 14 stained with Oil-Red-O. The photographs and micrographs shown are representative of 3 independent experiments. (B) RNA was isolated on day 14, and expression of <i>PPARG2</i>, <i>CEBPA</i>, <i>FABP4</i> and <i>LPL</i> was determined by RT-qPCR. Date presented were normalized to the value of DI treated cells. Significant differences are indicated by asterisks, *p<0.05, **p<0.01, ***p<0.001, n = 9. (C) Whole cell extracts were prepared and analyzed for FABP4 protein level by Western blotting. (D and E) Two-day post-confluent hMADS cells were treated for 15 min with 200 µM 8-pCPT-2′-O-Me-cAMP (007) or 100 µM 6-MB-cAMP (MB) or an equal volume of water (vehicle) in medium with Dex and Insulin. (D) GTP bound RAP1 was measured by a RAP1 activation pull-down assay as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#s4" target="_blank">Materials and Methods</a>”. (E) PKA activity in cell lysates was determined as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#s4" target="_blank">Materials and Methods</a>”. Significant differences relative to DI treated cells are indicated by asterisks, *p<0.05, **p<0.01, ***p<0.001, n = 4.</p

Carbaprostacyclin rescues adipocyte differentiation in the absence of other cAMP elevating agents.

<p>Two-day post-confluent hMADS cells were maintained in induction media with 0.86 µM insulin in the presence or absence of 1 µM Dex, 0.5 mM IBMX or 0.2 or 1 µM carbaprostacyclin (cPGI) as indicated from day 0 to day 3. Thereafter, 0.5 µM rosiglitazone was added until day 9. GPDH activities were determined on day 14.</p

Dominant negative Epac1 attenuates adipocyte differentiation of hMADS cells.

<p>hMADS cells were infected with retroviruses expressing a dominant-negative form of Epac1 (dnEpac1) or an empty vector and grow to confluence. (A) GTP-bound Rap1 was measured by a Rap1 activation pull-down assay. (B) Cells were induced to differentiate with Dex, insulin, IBMX and Rosi as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#s4" target="_blank">Materials and Methods</a>”. On day 14, cells were stained with Oil-Red-O and photographed. The photographs and micrographs shown are representative of 4 independent experiments. (C) <i>PPARG2</i>, <i>CEBPA</i>, <i>FABP4</i> and <i>LPL</i> mRNA levels were determined by RT-qPCR. Expression was normalized to cells transduced with empty vector. Significant differences are indicated by asterisks, **p<0.01, ***p<0.001, n = 6.</p

Expression of <i>Epac</i> and <i>Rap</i> mRNAs in hMADS cells.

<p>Cells were differentiated as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034114#s4" target="_blank">Materials and Methods</a>”. RNA was isolated on the indicated days. The levels of <i>RAPGEF3 (Epac1)</i>, <i>RAPGEF4 (Epac2)</i> mRNAs (A) and <i>RAP1A</i>, <i>RAP1B</i>, <i>RAP2A</i>, <i>RAP2B</i>, <i>RAP2C</i> mRNAs (B) were determined by RT-qPCR. Data presented in (A) were normalized to <i>RAPGEF3</i> on day 0; in (B) data were normalized to <i>RAP1B</i> on day0. Significant differences are indicated by asterisks, *p<0.05, **p<0.01, ***p<0.001, n = 8.</p