Cytological characterization of isolated sperm cells of perennial ryegrass (Lolium perenne L.).

The cytoplasmic content and the distribution of intramembrane particles (IMPs) of the plasma membrane of isolated sperm cells of perennial ryegrass (Lolium perenne L.) have been characterized using flow cytometry, transmission electron microscopy, confocal scanning laser microscopy and freeze-fracture studies. The isolated haploid sperm cells contain a variety of cell organelles with the exception of microtubules. Proplastids and plastids with starch were observed, although only rarely. Vacuoles containing remnants of organelles and stacked lamellae of endoplasmic reticulum with cytoplasmic inclusions were observed frequently, indicating that autophagy takes place. The number of mitochondria varies from 11 to 26 with an average of 17. Generally, the nucleus has a lobed shape and displays various interphasic stages of chromatin condensation. The analysis of the number of mitochondria and the nuclear state did not show evidence of sperm cell dimorphism. The cytological variability observed, could be explained by differences in developmental stages already present in vivo at the moment of isolation. No correlation between the number of mitochondria and the nuclear cross-sectioned area and/or the condensation state of the chromatin could be found. The density of intramembrane particles of the plasma membrane on the exoplasmic fracture face is more than twice that on the protoplasmic fracture face. That is the opposite of what was found for sporophytic cells of perennial ryegrass. These results are discussed in relation to the potential use of these cells for biotechnology and developmental studies

Analysis of biomarker expression in severe endometriosis and determination of possibilities for targeted intraoperative imaging

<p>Objective: To evaluate the expression of biomarkers in endometriotic tissue in order to determine the most promising molecules for targeted intraoperative imaging. Methods: Tissue samples were obtained from 18 patients with endometriosis. The intensity and pattern of expression of the following biomarkers were assessed by immunohistochemistry: C-X-C chemokine receptor type 4 (CXCR4), epithelial cell adhesion molecule (EpCAM), estrogen receptor (ER), folate receptor alpha (FR-alpha), hypoxia-inducible factor 1-alpha (HIF-1 alpha), progesterone receptor (PR), and vascular endothelial growth factor A (VEGF-A). The Target Selection Criteria scoring system was used to select the most promising biomarkers for intraoperative imaging. Results: Expression of OCCR4, EpCAM, ER, PR, and VEGF-A was scored as strong in endometriotic epithelium. Expression of FR-alpha was detected in 94.4% of samples, whereas HIF-1 alpha was expressed in just 5.6% of samples. Of note, CXCR4, ER, and VEGF-A were also expressed in surrounding healthy tissue, thus reducing the target-to-background ratio. Conclusion: Of the 7 biomarkets assessed in the present study, EpCAM, FR-alpha, and VEGF-A seem the most promising for targeted intraoperative imaging of endometriosis. (C) 2012 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.</p>