The daily activity pattern of small felids according to human cost of access.

<p>Activity patterns and overlap between: a) the activity pattern of ocelots at sites with low human cost of access (N = 481 activity records) vs. the activity at sites with high human cost of access (N = 937 activity records), b) the activity pattern of jaguarundis at sites with low human cost of access (N = 55 activity records) vs. the activity at sites with high human cost of access (N = 32 activity records), c) the activity pattern of margays at sites with low human cost of access (N = 16 activity records) vs. the activity at sites with high human cost of access (N = 25 activity records), d) the activity pattern of southern tiger cats at sites with low human cost of access (N = 79 activity records) vs. the activity at sites with high human cost of access (N = 85 activity records).</p

Activity and overlap of the small and medium felids.

<p>Temporal activity and degree of overlap (shaded area) among: a) ocelot and margay, overlap coefficient (Δ) = 0.89 with 95% CI = 0.82–0.95; b) jaguarundi and margay, Δ = 0.17 with 95% CI = 0.09–0.25; c) ocelot and jaguarundi, Δ = 0.21 with 95% CI = 0.17–0.25; d) margay and southern tiger cat, Δ = 0.57 with 95% CI = 0.47–0.66; e) jaguarundi and southern tiger cat, Δ = 0.63 with 95% CI = 0.56–0.69; f) ocelot and southern tiger cat, Δ = 0.60 with 95% CI = 0.53–0.66.</p

Activity of small felids according to the occupancy probability of ocelots.

<p>Activity patterns and overlap between: a) the daily activity pattern of jaguarondis at sites with low ψ of ocelots (N = 31 activity records) vs. the activity at sites with high ψ of ocelots (N = 90 activity records), b) the daily activity pattern of margays at sites with low ψ of ocelots (N = 14 activity records) vs. the activity at sites with high ψ of ocelots (N = 41 activity records), c) the daily activity pattern of southern tiger cat at sites with low ψ of ocelots (N = 72 activity records) vs. the activity at sites with high ψ of ocelots (N = 119 activity records).</p

Frequency of camera trap records and number of stations with presence (in parentheses) of medium and small felids at different landscape conditions in northern Misiones province, Argentina, where a camera trap survey was conducted between May 2013 and December 2014.

<p>Frequency of camera trap records and number of stations with presence (in parentheses) of medium and small felids at different landscape conditions in northern Misiones province, Argentina, where a camera trap survey was conducted between May 2013 and December 2014.</p

Effect of the human cost of access and habitat type on the occupancy probability of ocelots (ψ) in northern Misiones province, Argentina.

<p>Effect of the human cost of access and habitat type on the occupancy probability of ocelots (ψ) in northern Misiones province, Argentina.</p

Study area.

<p>a) Location of the Upper Parana Atlantic Forest (UPAF), Atlantic Forest (AF) and Misiones province, Argentina (in red). b) Location of the camera-trap stations during a survey conducted in the north of Misiones province between May 2013 and December 2014. The cameras were placed at continuous forest (CF, N = 53), fragmented forest (FF, N = 69) and pine plantations (PP, N = 62). c) Camera-trap stations placed in Misiones province between 2003 and 2016 (N = 586).</p

Occupancy probability of southern tiger cats (ψB).

<p>Location of the camera-trap stations (N = 184) with low (0–0.33, yellow), intermediate (0.34–0.66, orange), and high (0.67–1.00, red) occupancy probability of southern tiger cats according to the model of co-occurrence with the ocelot. Triangles = stations located in continuous forest, circles = forest fragment stations, squares = pine plantations.</p

Cellular distribution of endothelial and fibrotic markers involved in endotoxin-induced endothelial fibrosis.

<p>Representative images from experiments of vehicle-treated (A–H) or endotoxin (20 μg/mL LPS)-treated (I–P) ECs for 72 h. Endothelial markers CD31 or VE-cadherin (red), and the fibrotic markers α-SMA, or FSP-1 (green) were detected. In vehicle-treated cells: the box depicted in (A, C, E, and G) indicates the magnification shown in (B, D, F, and H), respectively. Arrows indicate CD31 (B, F) or VE-cadherin (D, H) labeling at the plasma membrane, whereas arrowheads indicate α-SMA (B, F) or FSP-1 (D, H) staining, indicating basal expression of fibrotic markers (B, D). In endotoxin-treated cells: the box depicted in (I, K, M, and O), indicates the magnification shown in (J, L, N, and P) respectively. Arrows indicate α-SMA (J, N) or FSP-1 (L. P), whereas arrowheads indicate CD31 (J, N) or VE-cadherin (L, P) staining from residual endothelial marker expression indicating EndMT. Nuclei were stained using DAPI. Bar scale represents 10 μm.</p

Endotoxin-induced endothelial fibrosis through changes in endothelial and fibrotic markers is dependent on TRPM7 expression.

<p>(A–B) TRPM7 expression downregulation by siRNA. ECs were transfected with a specific siRNA against the human TRPM7 isoform (siRNA-TRPM7) or a non-targeting siRNA (siRNA-CTRL). (A) Representative images from western blot experiments performed for detection of TRPM7 in cells transfected with siRNA-TRPM7 or siRNA-CTRL. (B) Densitometric analyses from several experiments, as shown in (A). Protein levels were normalized against tubulin, and the data are expressed relative to cells transfected with siRNA-CTRL condition. Statistical differences were assessed by student's t-test (Mann-Whitney). ***: <i>p</i><0.001. Graph bars show the mean ± SD (<i>N</i> = 3). (C–J) ECs were exposed to LPS for 72 h and protein expression was analyzed. (C–F) Representative images from western blot experiments performed for detection of endothelial markers CD31 (C) and VE-cadherin (VE-Cad) (D), and fibrotic markers α-SMA (E) and FSP-1 (F). (G–J) Densitometric analyses of the experiments shown in (C–F) respectively. Protein levels were normalized against tubulin and data are expressed relative to siRNA-CTRL transfected cells without endotoxin condition. Statistical differences were assessed by a one-way analysis of variance (ANOVA) (Kruskal–Wallis) followed by Dunn's post hoc test. *: <i>p</i><0.05 and **: <i>p</i><0.01 against to siRNA-CTRL transfected cells without endotoxin condition. NS: non-significant. Graph bars show the mean ± SD (<i>N</i> = 3–6).</p

Endotoxin-induced intracellular Ca²⁺ increase is mediated by TRPM7 in endothelial cells.

<p>(A) Representative time courses for normalized florescence in ECs loaded with Fura-2 and treated with vehicle (empty circles) or 20 μg/ml LPS (filled circles). The arrow represent the time for external solution change containing vehicle (empty circles) or LPS (filled circles). (B) Representative time courses for normalized florescence in ECs transfected with non-targeting siRNA (siRNA-CTRL, filled circles) or with siRNA for TRPM7 (siRNA-TRPM7, empty circles) and loaded with Fura-2. The arrow indicates the time for LPS application. (C) Summary bar graph for maximal amplitude of normalized florescence in wild type ECs (WT) or transfected with siRNAs. (D) Summary of the data for the time constant (τ) of calcium return to basal levels, obtained after fitting the data to a single exponential for wild type ECs (WT) or transfected with siRNAs. (E–H) Endothelial cells were incubated in the absence (−) or presence (+) of 20 μg/mL endotoxin and the calcium overloading was evaluated by means of the Ca²⁺-sensitive fluorescent dye fluo-4. (E) ECs were transfected with siRNA-TRPM7 and siRNA-CTRL and incubated in the absence (−) or presence (+) of endotoxin. (F) ECs were preincubated with 0.5 mM MCI-186, 1 mM NAC or 2 mM GSH for 1 hr and then incubated in the absence (−) or presence (+) of endotoxin. (G) ECs were transfected with siRNA-TRPM7 and siRNA-CTRL and incubated in the absence (−) or presence (+) of 10 μM H₂O₂. (H) ECs were preincubated with 5 mM L-NAME, 1 mM L-NMMA, 50 μM cobinamide, or 500 μM PTIO for 1 hr and then incubated in the absence (−) or presence (+) of endotoxin. Statistical differences were assessed by a one-way analysis of variance (ANOVA) (Kruskal–Wallis) followed by Dunn's post hoc test. *: <i>p</i><0.05; **: <i>p</i><0.01 against vehicle-treated condition. Graph bars show the mean ± SD (<i>N</i> = 3–4).</p