363 research outputs found

    Uncovering the physiological mechanisms underlying the roe deer (Capreolus capreolus) testicular cycle: Analyses of gelatinases and VEGF patterns and correlation with testes weight and testosterone

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    The roe deer (Capreolus capreolus) represents a spontaneous model of testicular inactivation: During winter, bucks show a suspension of spermatogenesis that starts again in spring and peaks during the breeding season (July\u2013August). The underlying mechanisms to the regulation of the cyclic testicular changes are still not fully clear but seem to be imputable to the spermatogenic cell line since other testicular cell populations remain stable without apoptotic phenomena. The aim of the study was to investigate apoptosis, gelatinases (MMP2 and 9), their inhibiting factors (TIMP 1-2), and two isoforms of vascular endothelial growth factor (VEGF121 and 165) with its receptors (VEGFR1-2) in testes collected during pre-and post-rut periods, and to correlate them with testicular weight (TW) and testosterone (TEST). Testes from 18 adult sexually mature bucks were collected in Bologna Apennines (Italy). Samples were weighed and parenchyma collected. Radioimmunoassay, real-time PCR, and zymography were performed. The results showed a post-rut decrease in TW and TEST and an increase in proMMP2, also highlighting a correlation between the gelatinases and the testicular functionality. The VEGF pattern did not show modifications nor correlation with TW and TEST. Overall, gelatinases and their inhibitors, described herein for the first time in roe deer testes, seem to play an important role in the testicular cycle

    Ex vivo effect of vascular wall stromal cells secretome on enteric ganglia

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    BACKGROUND Mesenchymal stromal cell (MSC)-based therapy is currently under study to treat inflammatory bowel diseases. MSC bioactive products could represent a valid alternative to overcome issues associated with systemic whole-cell therapies. However, MSC anti-inflammatory mechanisms differ between rodents and humans, impairing the reliability of preclinical models. AIM To evaluate the effect of conditioned medium (CM) derived from porcine vascular wall MSCs (pVW-MSCs) on survival and differentiation of porcine and Guinea pig enteric ganglia exposed to lipopolysaccharide (LPS). METHODS Primary cultures of enteric ganglia were obtained by mechanic and enzymatic digestion of ileum resections from Guinea pigs (Cavia porcellus) (GPEG) and pigs (Suus scrofa) (PEG). pVW-MSCs were derived by enzymatic digestion from vascular wall resections of porcine aorta and tested by immunoflowcytometry for MSC immune profile. Enteric ganglia were treated with increasing concentrations of LPS, CM derived by pVW-MSCs or a combination of CM and LPS 1 \u3bcg/mL. Cell count and morphometric analysis of HuD positive neurons and glial fibrillary acidic protein positive glial cells were performed by immunofluorecent staining of cultured ganglia. RESULTS PEG showed a higher number of neurons compared to GPEG. Overall, CM exerted a protective role on LPS-treated enteric ganglia. CM in combination with LPS increased the number of glial cells per ganglion in both cultures evoking glial cells differentiation in porcine cultures. CONCLUSION These findings suggest an immunomodulating activity of pVW-MSCs mediators on the enteric nervous system in inflammatory conditions

    Preliminary assessment of the mucosal toxicity of tea tree (Melaleuca alternifolia) and rosemary (rosmarinus officinalis) essential oils on novel porcine uterus models

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    Antimicrobial resistance, an ever-growing global crisis, is strongly linked to the swine production industry. In previous studies, Melaleuca alternifolia and Rosmarinus officinalis essential oils have been evaluated for toxicity on porcine spermatozoa and for antimicrobial capabilities in artificial insemination doses, with the future perspective of their use as antibiotic alternatives. The aim of the present research was to develop and validate in vitro and ex vivo models of porcine uterine mucosa for the evaluation of mucosal toxicity of essential oils. The in vitro model assessed the toxicity of a wider range of concentrations of both essential oils (from 0.2 to 500 mg/mL) on sections of uterine tissue, while the ex vivo model was achieved by filling the uterine horns. The damage induced by the oils was assessed by Evans Blue (EB) permeability assay and histologically. The expression of ZO-1, a protein involved in the composition of tight junctions, was assessed through immunohistochemical and immunofluorescence analysis. The results showed that low concentrations (0.2\u20130.4 mg/mL) of both essential oils, already identified as non-spermicidal but still antimicrobial, did not alter the structure and permeability of the swine uterine mucosa. Overall, these findings strengthen the hypothesis of a safe use of essential oils in inseminating doses of boar to replace antibiotics

    Constitutive and LPS-stimulated secretome of porcine Vascular Wall-Mesenchymal Stem Cells exerts effects on in vitro endothelial angiogenesis

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    Background: MSCs secretome is under investigation as an alternative to whole-cell-based therapies, since it is enriched of bioactive molecules: growth factors, cytokines and chemokines. Taking into account the translational value of the pig model, the leading aim of the present paper was to characterize the secretome of porcine Vascular Wall-Mesenchymal Stem Cells (pVW-MSCs) and its change in presence of LPS stimulation. Moreover, considering the importance of angiogenesis in regenerative mechanisms, we analysed the effect of pVW-MSCs secretome on in vitro angiogenesis. Results: Our results demonstrated that conditioned medium from unstimulated pVW-MSCs contained high levels of IL-8, GM-CSF, IFN-\u3b3 and other immunomodulatory proteins: IL-6 IL-18 IL-4 IL-2 IL-10. LPS modulates pVW-MSCs gene expression and secretome composition, in particular a significant increase of IL-6 and IL-8 was observed; conversely, the amount of GM-CSF, IFN-\u3b3, IL-2, IL-4, IL-10 and IL-18 showed a significant transient decrease with the LPS stimulation. Conditioned medium from unstimulated pVW-MSCs induced in vitro endothelial angiogenesis, which is more evident when the conditioned medium was from LPS stimulated pVW-MSCs. Conclusions: The lines of evidence here presented shed a light on possible future application of secretome derived by pVW-MSCs on research studies in translational regenerative medicine

    Some factors influencing quality of spontaneous or induced sputum for inflammatory cell analysis

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    predict the quality of the sputum samples obtained in a large group of asthmatic subjects. Methods. We compared the presence of sputum productive cough in the days preceding the test, easiness in expectoration during the test, and sputum macroscopic aspect (presence of visible plugs) with the quality of slides obtained from sputum processing. We also monitored changes in the quality in patients who repeated sputum collection several times, comparing those whose first sample was adequate with those whose first sample was inadequate. We analysed 547 sputum samples obtained from 238 asthmatic patients. Sputum was processed using the whole sample method. Results. Patients with productive cough in the days preceding the test and easy expectoration during the test produced a higher percentage of adequate samples than those without productive cough (86% vs 76%, p=0.01) and with difficulty in expectoration (85% vs 63%, p=0.0001). "Good" macroscopic samples were associated with better quality of slides (91% vs 38%, p=0.0001). Patients with inadequate first sample (n=40) had a higher percentage of inadequate samples (55%) in the subsequent tests than patients (n=115) with adequate first sample (8%). Conclusions. Patients with increased airway secretions in the days preceding the test, easy expectoration and "good" macroscopic aspect of the sputum are more likely to produce sputum sample adequate for inflammatory cell analysis. If the first sputum sample is adequate, subsequent samples are very likely to be adequate as well. If the first sputum sample is inadequate, the quality of subsequent samples cannot be predicted, since there are similar probabilities of having adequate or inadequate samples

    Effective Lagrangian Approach to the Theory of Eta Photoproduction in the N(1535)N^{*}(1535) Region

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    We investigate eta photoproduction in the N(1535)N^{*}(1535) resonance region within the effective Lagrangian approach (ELA), wherein leading contributions to the amplitude at the tree level are taken into account. These include the nucleon Born terms and the leading tt-channel vector meson exchanges as the non-resonant pieces. In addition, we consider five resonance contributions in the ss- and uu- channel; besides the dominant N(1535)N^{*}(1535), these are: N(1440),N(1520),N(1650)N^{*}(1440),N^{*}(1520),N^{*}(1650) and N(1710)N^{*}(1710). The amplitudes for the π\pi^\circ and the η\eta photoproduction near threshold have significant differences, even as they share common contributions, such as those of the nucleon Born terms. Among these differences, the contribution to the η\eta photoproduction of the ss-channel excitation of the N(1535)N^{*}(1535) is the most significant. We find the off-shell properties of the spin-3/2 resonances to be important in determining the background contributions. Fitting our effective amplitude to the available data base allows us to extract the quantity χΓηA1/2/ΓT\sqrt{\chi \Gamma_\eta} A_{1/2}/\Gamma_T, characteristic of the photoexcitation of the N(1535)N^{*}(1535) resonance and its decay into the η\eta-nucleon channel, of interest to precise tests of hadron models. At the photon point, we determine it to be (2.2±0.2)×101GeV1(2.2\pm 0.2)\times 10^{-1} GeV^{-1} from the old data base, and (2.2±0.1)×101GeV1(2.2\pm 0.1) \times 10^{-1} GeV^{-1} from a combination of old data base and new Bates data. We obtain the helicity amplitude for N(1535)γpN^{*}(1535)\rightarrow \gamma p to be A1/2=(97±7)×103GeV1/2A_{1/2}=(97\pm 7)\times 10^{-3} GeV^{-1/2} from the old data base, and A1/2=(97±6)×103GeV1/2A_{1/2}=(97\pm 6)\times 10^{-3} GeV^{-1/2} from the combination of the old data base and new Bates data, compared with the results of the analysis of pion photoproduction yielding 74±1174\pm 11, in the same units.Comment: 43 pages, RevTeX, 9 figures available upon request, to appear in Phys. Rev.

    A direct search for the CP-violating decay Ks->3p^0 with the KLOE detector at DAFNE

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    We have searched for the decay Ks->3p^0 with the KLOE experiment at DAFNE using data from e^+ e^- collisions at a center of mass energy W= m(phi) for an integrated luminosity L=450 pb^-1. The search has been performed with a pure Ks beam obtained by tagging with Kl interactions in the calorimeter and detecting six photons. We find an upper limit for the branching ratio of 1.2x10^-7 at 90% C.L.Comment: 15 pages, 6 figures. To be submitted to Physics Letter
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