5 research outputs found

    European Society for Sexual Medicine Consensus Statement on the Use of the Cavernous Nerve Injury Rodent Model to Study Postradical Prostatectomy Erectile Dysfunction

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    Introduction: Rodent animal models are currently the most used in vivo model in translational studies looking into the pathophysiology of erectile dysfunction after nerve-sparing radical prostatectomy. Aim: This European Society for Sexual Medicine (ESSM) statement aims to guide scientists toward utilization of the rodent model in an appropriate, timely, and proficient fashion. Methods: MEDLINE and EMBASE databases were searched for basic science studies, using a rodent animal model, looking into the consequence of pelvic nerve injury on erectile function. Main outcome measures: The authors present a consensus on how to best perform experiments with this rodent model, the details of the technique, and highlight possible pitfalls. Results: Owing to the specific issue—basic science—Oxford 2011 Levels of Evidence criteria cannot be applied. However, ESSM statements on this topic will be provided in which we summarize the ESSM position on various aspects of the model such as the use of the Animal Research Reporting In Vivo Experiments guideline and the of common range parameter for nerve stimulation. We also highlighted the translational limits of the model. Conclusion: The following statements were formulated as a suggestive guidance for scientists using the cavernous nerve injury model. With this, we hope to standardize and further improve the quality of research in this field. It must be noted that this model has its limitations

    Box plot of the quantitation of expression of WNT4, MMP7, CD1 and c-MYC in PeCa tissue classified according to pathological grade.

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    <p>97% of the samples used in this study were classified as grade I or II. The data presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0124395#pone.0124395.t001" target="_blank">Table 1</a> was segregated into expression according to pathological grades and plotted as box plots. Filled circles are 99% value and horizontal line within the box is the median value. Protein expression (mean gray value per core) in control samples (green, n = 37); malignant tumor sample classified as grade I by a pathologist (blue, n = 82) or grade II (purple, n = 18). All proteins showed a significant difference in expression between control and malignant tumor samples. Significance of difference was measured using Mann-Whitney U test: * = p<0.001 and <sup>§</sup> = p<0.01 between control and malignant grade I or grade II samples or within grade I and grade II. There was no significant difference in protein expression for any marker between grade I and grade II samples.</p

    Box plot of differential pattern for WNT4, MMP7, CD1 and c-MYC co-expression in control and malignant penile tissue.

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    <p>Quantitative co-localization was performed from high magnification images (e.g. as represented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0124395#pone.0124395.g003" target="_blank">Fig 3</a>) and image files used for quantitation as described in materials and methods in WCIF ImageJ software co-localization plugin to measure the Pearson coefficient of co-localization. 35 individual tissue cores that included control (gradient from centre ) and malignant grade I (gradient from left to right ) and grade II (gradient from right to left ) were used for each fluorophore to calculate the co-localization of expression of WNT4, MMP7, CD1 and c-MYC, generating 12 comparisons: CD1 and WNT4 (orange/blue), MMP7 and WNT4 (green/blue), c-MYC and WNT4 (pink/blue), CD1 and c-MYC (orange/pink), MMP7 and c-MYC (green/pink), MMP7 and CD1 (green/orange). Significance of difference in the Pearson coefficient of co-localization between control and grades I and II (*0.02–0.04) and between grade I and grade II (§ p<0.04, range 0.04–0.0001) samples was calculated using the Mann-Whitney U test; boxes showing comparisons without symbols are not significantly different.</p

    Representative images from Zeiss AxioScan Z1 slide scanner.

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    <p>Composite, overlay, of four fluorophores (FITC, Alexa Fluor-405, Cy3 and Cy5 for MMP7, Wnt4, CD1 and c-MYC, respectively) in (A) PeCa tissue cores (core B2 of PE241) and (B) control (core D5 of PE241, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0124395#sec002" target="_blank">methods</a>). C-F are individual images used to construct the composite of PeCa tissue core, for MMP7 (C, green), Wnt4 (D, blue), CD1 (E, orange) and c-MYC (F, red). G-J are individual images used to construct the composite of control tissue core, for MMP7 (G, green), Wnt4 (H, blue), CD1 (I, orange) and c-MYC (J, red). Quantitative analysis was performed on the non-enhanced, original, gray images for which all settings were identical in all tissue core images analyzed. Scale bar = 250 μm.</p
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