Blood inflammatory indices in goats around kidding

AbstractThe transition period of goats is often characterized by serious metabolic problems, mainly before parturition. These troubles are related to negative energy balance status, however all causes are not totally defined. To improve the knowledge about pathogenesis in this phase we have monitored the changes of some blood indices of lipomobilization and inflammation. Six blood samples were collected from 10 primiparous and 25 multiparous "Camosciata delle Alpi" goats after morning milking. Samples were collected around 20 and 7 days before parturition and on days 0, 3, 6 and 12 of lactation. Albumin, total protein, haptoglobin, ceruloplasmin, total cholesterol, NEFA, β-OH-butyrate (BHB), Ca, Mg and Zn were determined. Goats were grouped according to their BHB level before parturition: low (≤0.6 mmol/l; LOB), average (0.6÷1.09; AVB) and high (≥1.09 mmol/l; HIB) level. Furthermore, the AVB group was divided according to plasma haptoglobin level before parturition: low (<0.5 g/l) or high. The statistical..

Comparative analyses of respiratory rates induced by different substrates among <i>A</i>. <i>aegypti</i> sexes.

<p>Oxygen consumption rates from isolated mitochondria (A-C) and whole permeabilized flight muscle (D and E) from females (solid bars) and males (hatched bars) were plotted from values shown in Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.t003" target="_blank">3</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.t004" target="_blank">4</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.s010" target="_blank">S2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.s011" target="_blank">S3</a>. Data are expressed as mean ± SD of at least six different experiments. Comparisons between groups were done by Student´s t- test. Figure (A): ^<i>a</i><i>p</i><0.005 and ^<i>b</i><i>p</i><0.05 relative to their equivalent metabolic state in female. Figure (D): ^<i>a</i><i>p</i><0.0001 and ^<i>b</i><i>p</i><0.05 relative to their equivalent metabolic state in female. Figure (E): ^<i>a</i><i>p</i><0.001 and ^<i>b</i><i>p</i><0.05 relative to their equivalent metabolic state in female.</p

Schematic representation of substrate utilization pathways driving respiration and O2•¯ formation in <i>A</i>. <i>aegypti</i> flight muscle mitochondria.

<p>The dehydrogenases directly involved on mitochondrial electron transfer from nutrient oxidation to respiration are depicted in their respective colors utilized throughout this work, as following: complex I (light green), ProDH (dark green), G3PDH (red) and ETF:QOR (blue). The contribution of dehydrogenases to respiration are represented by their boxes, fonts, and lines sizes. Electron leak and O₂^•¯ formation induced by different substrates are represented by steam clouds, obeying the same color and size pattern described for dehydrogenases. Noteworthy, the steam cloud location in this scheme does not represent the exact site of O₂^•¯ production, since we were unable to precisely define these sites in this work. CACT, carnitine-acylcarnitine transferase; CPT2, carnitinepalmitoyl transferase 2; palm-CoA, palmitoyl-CoA; αKG, alpha-ketoglutarate; Δ1PC, Δ-1-pyrroline-5-carboxylate; DHAP, dihydroxyacetone phosphate; PDH, pyruvate dehydrogenase; IMS, intermembrane space; MM, mitochondrial matrix.</p

Topology of H₂O₂ formation in <i>A</i>. <i>aegypti</i> flight muscle mitochondria.

<p>Values were expressed as mean ± SD of pmol hydrogen peroxide produced/min/mg protein in three different mitochondrial metabolic states using: 10 mM pyruvate + 10 mM proline (Pyr+pro), 20 mM <i>sn</i> glycerol-3 phosphate (G3P) or 10 μM palmitoylcarnitine + 5 mM malate (PC+Mal) followed by 2 mM ADP, 4 μg/mL oligomycin, 2 μM FCCP, 0.5 μM rotenone, 2.5 μg/mL antimycin A. Statistical analyses within groups were performed by using Mann Whitney test (superscript letters), Student t test (superscript symbols) or Kruskal-Wallis test followed by <i>a posteriori</i> Dunn´s test (superscript open squares, □ or closed circles ^●).</p><p>^a, p = 0.0079, relative to female Pyr+pro FCCP;</p><p>^b, p = 0.0079, relative to male Pyr+pro FCCP;</p><p>^c, p = 0.0079, relative to female PC+Mal FCCP;</p><p>^d, p = 0.03, relative to male PC+Mal FCCP;</p><p>*, p<0.0001 relative to female G3P Rot;</p><p>#, p<0.0001 relative to male G3P Rot.</p><p>^□, p = 0.004, relative to male and female Pyr+pro and G3P antimycin.</p><p>^●, p = 0.005, relative to female Pyr+pro and G3P and male G3P antimycin.</p><p>Topology of H₂O₂ formation in <i>A</i>. <i>aegypti</i> flight muscle mitochondria.</p

Preference towards proline oxidation in <i>A</i>. <i>aegypti</i> female mitochondria.

<p>Oxygen consumption rates from isolated mitochondria (A-E) and whole permeabilized flight muscle (F-H) from females (solid bars) and males (hatched bars) were calculated from values shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.g003" target="_blank">Fig. 3</a>. Data are expressed as mean ± SD of at least seven different experiments. Comparisons between groups were done by Student´s t tests. Figure (B): ^<i>b</i><i>p</i><0.001 relative to female; Figure (G): ^<i>a</i><i>p</i><0.0001 relative to female; Figure (H): ^<i>b</i><i>p</i><0.038 relative to female.</p

Contribution of different substrates to mitochondrial H₂O₂ production in <i>A</i>. <i>aegypti</i> flight muscle.

<p>Values were expressed as mean ± SD of pmol hydrogen peroxide produced/min/mg protein in two different mitochondrial metabolic states using: 10 mM pyruvate + 10 mM proline (Pyr+pro), 20 mM <i>sn</i> glycerol-3 phosphate (G3P), or 10 μM palmitoylcarnitine + 5 mM malate (PC+Mal) followed by 2 mM ADP, 4 μg/mL oligomycin (Oligo), and 2 μM FCCP. For all G3P experiments, measurements were carried out after addition of 0.5 μM rotenone. Statistical analyses were carried out between the groups of different substrates and mitochondrial metabolic state within the same sex, and were performed by using Kruskal-Wallis test followed by <i>a posteriori</i> Dunn´s test (indicated by superscript letters). Significant difference in “Oligo” was</p><p>^<i>a</i><i>p = 0</i>.<i>0052</i>, relative to Pyr+pro and G3P,</p><p>^<i>b</i><i>p = 0</i>.<i>0045</i>, relative to G3P. Significant differences in “FCCP” was;</p><p>^{<i>c </i>}<i>p = 0</i>.<i>0065</i>, relative to G3P;;</p><p>^<i>d</i><i>p = 0</i>.<i>0016</i>, relative to G3P. Analyses were also conducted between the two mitochondrial metabolic states (oligo vs. FCCP) within the same substrate and sex and were performed by using Mann Whitney test (indicated by superscript symbols). Significant differences in G3P were ** <i>p</i><0.0001 relative to female FCCP;</p><p>* <i>p</i> = 0.016 relative to male FCCP. Significant difference in PC+Mal were</p><p># <i>p</i> = 0.03 relative to female FCCP;</p><p>Contribution of different substrates to mitochondrial H₂O₂ production in <i>A</i>. <i>aegypti</i> flight muscle.</p

Mitochondrial Physiology in the Major Arbovirus Vector <i>Aedes aegypti</i>: Substrate Preferences and Sexual Differences Define Respiratory Capacity and Superoxide Production

<div><p>Adult females of <i>Aedes aegypti</i> are facultative blood sucking insects and vectors of Dengue and yellow fever viruses. Insect dispersal plays a central role in disease transmission and the extremely high energy demand posed by flight is accomplished by a very efficient oxidative phosphorylation process, which take place within flight muscle mitochondria. These organelles play a central role in energy metabolism, interconnecting nutrient oxidation to ATP synthesis, but also represent an important site of cellular superoxide production. Given the importance of mitochondria to cell physiology, and the potential contributions of this organelle for <i>A</i>. <i>aegypti</i> biology and vectorial capacity, here, we conducted a systematic assessment of mitochondrial physiology in flight muscle of young adult <i>A</i>. <i>aegypti</i> fed exclusively with sugar. This was carried out by determining the activities of mitochondrial enzymes, the substrate preferences to sustain respiration, the mitochondrial bioenergetic efficiency and capacity, in both mitochondria-enriched preparations and mechanically permeabilized flight muscle in both sexes. We also determined the substrates preferences to promote mitochondrial superoxide generation and the main sites where it is produced within this organelle. We observed that respiration in <i>A</i>. <i>aegypti</i> mitochondria was essentially driven by complex I and glycerol 3 phosphate dehydrogenase substrates, which promoted distinct mitochondrial bioenergetic capacities, but with preserved efficiencies. Respiration mediated by proline oxidation in female mitochondria was strikingly higher than in males. Mitochondrial superoxide production was essentially mediated through proline and glycerol 3 phosphate oxidation, which took place at sites other than complex I. Finally, differences in mitochondrial superoxide production among sexes were only observed in male oxidizing glycerol 3 phosphate, exhibiting higher rates than in female. Together, these data represent a significant step towards the understanding of fundamental mitochondrial processes in <i>A</i>. <i>aegypti</i>, with potential implications for its physiology and vectorial capacity.</p></div

Schematic representation of the electron transport system, showing the sites of action of oxidative phosphorylation (OXPHOS) modulators (brown), the different substrates utilized throughout this study (pyruvate and proline, green; glycerol 3 phosphate, red; palmitoylcarnitine and malate, blue) and the known sites of superoxide (O2•¯) production (purple).

<p>Schematic representation of the electron transport system, showing the sites of action of oxidative phosphorylation (OXPHOS) modulators (brown), the different substrates utilized throughout this study (pyruvate and proline, green; glycerol 3 phosphate, red; palmitoylcarnitine and malate, blue) and the known sites of superoxide (O2•¯) production (purple).</p

Contribution of different electron leak sites to H₂O₂ generation in isolated <i>A</i>. <i>aegypti</i> flight muscle mitochondria.

<p>The contribution of site I_F, ProDH+other dehydrogenases, G3PDH+other dehydrogenases and ETF:QOR+other dehydrogenases sites to H₂O₂ generation in <i>A</i>. <i>aegypti</i> mitochondria isolated from females (A, solid colors) and males (B, hatched bars) were calculated from data shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.t008" target="_blank">Table 8</a>. Data are expressed as mean ± SD of at least five different experiments. Comparisons between groups were done by ANOVA and <i>a posteriori</i> Tukey´s tests. Figure (A): ^<i>a</i><i>p</i><0.001 relative to I_F (Pyr+pro); ^<i>b</i><i>p</i><0.001 relative to ETF:QOR+other dehydrogenases; ^<i>c</i><i>p</i><0.001 relative to I_F (PC+Mal); ^<i>d</i><i>p</i><0.05 relative to G3PDH+other dehydrogenases; Figure (B): ^<i>a</i><i>p</i><0.001 relative to I_F (Pyr+pro); ^<i>b</i><i>p</i><0.001 relative to ETF:QOR+other dehydrogenases; ^<i>c</i><i>p</i><0.001 relative to I_F (PC+Mal); ^<i>d</i><i>p</i><0.05 relative to G3PDH+other dehydrogenases.</p

Sexual differences in the contribution of different electron leak sites to H₂O₂ generation in isolated <i>A</i>. <i>aegypti</i> flight muscle mitochondria.

<p>The contribution of site I_F, ProDH+other dehydrogenases, G3PDH+other dehydrogenases and ETF:QOR+other dehydrogenases sites were calculated in <i>A</i>. <i>aegypti</i> mitochondria isolated from females (solid colors) and males (hatched bars) from data shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120600#pone.0120600.t008" target="_blank">Table 8</a>. Data are expressed as mean ± SD of at least five different experiments. Comparisons between groups were done by Mann Whitney test. Figure (D): ^<i>a</i><i>p</i><0.01 relative to female I_F (PC+Mal).</p