Increasing evidence shows that atherosclerosis is a low-grade chronic inflammatory disease. The interaction between the arterial wall and circulating blood cells like monocytes and T lymphocytes is an important step in this inflammatory process. This interaction is mediated via transmembrane glycoprotein receptors known as cellular adhesion molecules (CAM). The expression of these molecules is provoked by a variety of cardiovascular risk factors and is under the control notably of inflammatory cytokines, hemodynamic factors, bacterial endotoxins. Recently, data from large prospective studies have shown that an increased concentration of CAM predicts an increased cardiovascular risk. There is an increasing interest in CAM as a new therapeutic target in atherosclerosis.
In the present studies atrial natriuretic factor (ANF) was characterized immunocytochemically in the reproductive tract of immature female rats, and changes of ANF levels in response to different hormonal conditions were demonstrated. Administration of pregnant mare serum gonadotropin (PMSG) to immature animals has shown to be a useful method to synchronize growth, differentiation and atresia of ovarian follicles. ANF immunoreactivity was investigated in rat uterus and oviduct during follicular growth and estrogenic dominance (48 h after PMSG treatment) and during follicular atresia and progesterone dominance (96 h after PMSG treatment). Our immunocytochemical results showed that in rat uterus ANF was localized in endometrial mucosal and glandular epithelium and smooth muscle cells of the myometrium. In the oviduct ANF immunoreactivity was observed in mucosal cells and muscle layers. Immunocytochemical staining patterns and Western blot analysis revealed that ANF levels in rat uterus and oviduct are modulated by the hormonal status. ANF immunoreactivity was elevated during estrogenic dominance (48 h after PMSG) in uterus and oviduct. However, during progesterone dominance (96 h after PMSG) elevation of ANF immunoreactivity was observed in the uterus only. These results raise the possibility that ANF expression in rat oviduct is positively controlled by estrogen and negatively by progesterone. ANF staining in uterus during progesterone phase provides evidence that both estrogen and progesterone regulate ANF levels in uterus. The observed staining patterns indicate that ANF may have intracellular functions as well as a role in priming the extracellular environment. Accordingly, the possibility that ANF might be an important regulatory molecule for autocrine/paracrine communication within the female reproductive tract should be considered.
<p>PDF file - 557KB, Supplementary Methods Suppl. Table 1. Primers used for real time and conventional RT-PCR . Suppl. Figure S1. RhoE expression is down-regulated in skin SCC tumors. Suppl. Figure S2. Notch activation does not induce expression of members of the Rho GTPase family of proteins. Suppl. Figure S3. Endogenous activation of Notch signaling induces the expression of Hey2. Suppl. Figure S4. Depletion of RhoE inhibits downstream Notch target genes. Suppl. Figure S5. Real-time RT-PCR analysis of mRNA isolated from primary keratinocytes. Suppl. Figure S6. Levels of RhoE and Keratin1 are increased during keratinocyte differentiation. Suppl. Figure S7. RhoE ablation suppresses growth and commitment to differentiation. Suppl. Figure S8. Negative control for RhoE and Notch1 staining in SCC4 cells. Suppl. Figure S9. Depletion of RhoE abrogates the binding between NIIC and importin Beta1.</p>
SUMMARYA monoclonal antibody (Mab 2a8) has been generated against a DNA tight binding protein of eukaryotic cells. Its distribution was investigated in salivary gland polytene chromosomes of two Chironomid species, Endochironomus tendens and Glyptotendipes gripekoveni, by immunofluorescence. The results indicate: 1. Mab 2a8 is distributed in some distinct condensed bands along the entire lenght of chromosomes; 2. The banding pattern obtained with fluorescent antibody does almost strictly correspond to some Hoechst positive bands and to constitutive heterochromatin sites established by «C» banding method. Other condensed (Hoechst positive) bands on chromosomes were Mab 2a8 immunonegative. 3. No immunofluorescent was observed in genetically active regions of polytene chromosomes (puffs, Balbiani rings and nuclear organizer). The mode of distribution in defined bands of polytene chromosomes suggests the residence of this protein in polytene chromosomes, whose presence appears to be well in correlation with the centromere and intercalary heterochromatin regions. This protein exibits preferential binding to specific chromosome sites containing DNA sequences preferably involved in a specific interaction with target protein. This protein is probably conserved during the evolution being represented in species distant from a phylogenetical point of view (mammalian cells, polytene chromosomes of Chironomid).
A procedure was designed for purification of a human seminal plasma-specific antigen (HSP-antigen) identified by a monoclonal antibody produced in this laboratory (mAb 4E6). Pooled human seminal plasma was fractionated by consecutively applied methods: affinity chromatography on Lentil lectin sepharose, gel chromatography on Ultrogel AcA 34 and immunoaffinity on mAb 4E6 coupled CNBr-Sepharose 4B. The antigen-containing fraction obtained after this procedure was proved to be homogeneous when analysed by electrophoresis in polyacrylamide gel. After the consecutive purification procedures the degree of purification was 128 times as compared to the starting material. Electrophoretic analysis of the purified 4E6 antigen under reducing conditions showed that it consisted of 3 polypeptide subunits with molecular weight 70 kDa, 64 kDa and 60 kDa respectively. On the basis of the data obtained from competitive ELISA testing of sera from infertile patients it has been suggested that the identified antigen may be involved in pathogenesis of immunologic infertility.
Human endometrium is an object of extensive restructuring and remodeling during the female reproductive life and it is quite tempting to assume that these periodic changes happen with the participation of cells that should have the basic characteristics of multipotent cells. The aim of this study was to search for the presence of cells with plastic adherence, clonogenicity, and differentiation in human endometrium. To this end, human endometrial stromal cells were cultured in vitro for more than 15 passages. Flow cytometry analysis of the cultured cells showed that they were positive for CD29, CD73 and CD90, which are considered to be the markers of cells with mesenchymal origin. The cells were negative for the hematopoietic cell markers (CD45, CD34, CD14, CD3, CD19, CD16/56, and HLA-DR). Further, it was shown that the cultured cells had 15% clonogenic efficiency and could be induced to differentiate into adipogenic cells containing typical lipid-rich vacuoles. These results demonstrate that the human endometrium contains a low number of cells with the characteristics of endometrial stromal stem/progenitor cells, which seem to belong to the family of the mesenchymal stem cells. It can be speculated that these cells are engaged into the monthly restructuring and remodeling of human endometrium.
