Endometriosis is a chronic gynecological disorder defined as the presence of endometrial tissue within extra-uterine sites. The primary symptoms are infertility and chronic pain. The inflammatory environment and aberrant immune responses in women with endometriosis may be directly associated with the initiation and progression of endometriotic lesions. In the present study, the secretion of inflammatory cytokines was evaluated in cultures of primary endometrial cells (ECs) isolated from the endometrium of women with and without endometriosis. The presence of endometriotic cells leads to alterations in the secretory profile of healthy ECs. The expression of the inflammatory cytokines interleukin (IL)-6 and IL-8 was significantly increased in endometriotic and co-cultured cells compared with healthy ECs. IL-6 expression was strongly correlated with IL-8 expression in endometriotic cells. IL-1β expression was increased on day 10 of co-culture to 48.30 pg/ml and may be associated with the long-term co-culture, rather than IL-6 and IL-8 expression. IL-6 expression was strongly correlated with cell number, whereas IL-8 expression was moderately correlated with cell number. Additionally, it was observed that co-cultured cells exhibited a different population of cells, with expression of the mesenchymal stem cell marker cell surface glycoprotein MUC18, indicating a putative role of endometrial mesenchymal stem cells in the secretion of cytokines and disease development. These results indicate a predominant role of primary endometriotic cells in the secretion of cytokines, which contributes to the disrupted peritoneal and endometrial environment observed in the women with endometriosis.

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This study evaluated the in vitro antiviral activity of propolis and Baccharis sp. extracts on three animal herpesviruses (bovine, equine and swine). The propolis samples were produced by two species of bees. There was red and green propolis, which came from africanized Apis melifera, and a third type obtained from a native bee species, Tetragonisca angustula (jatai). The Baccharis extracts were obtained from four different species: B. oblongifolia, B. burchellii, B. dracunculifolia and B. uncinella. The maximum non-toxic concentration of the extracts was determined when no visible morphological changes were observed on the cells. These non-toxic concentrations were used in the antiviral tests. Antiviral activity was evaluated using a reduction assay of the cytopathic effect, which calculated the difference between treated and control virus titer by statistical analysis. Red propolis was active against the three herpesviruses and green propolis showed inhibition against the equine and swine herpesviruses. Conversely, jataí propolis showed no antiviral activity. Most extracts coming from male and female individuals of all of the Baccharis species showed antiviral activity against bovine and swine herpesviruses. Only the extract of the female specimen of B. oblongifolia was an inhibitor against equine herpesvirus.

Background and ObjectivesGerbich (GE) blood group system carries high‐frequency antigens and the absence of them leads to rare phenotypes: GE:−2,3,4, GE:−2,−3,4 and GE:−2,−3,−4. Their serological differentiation is limited and misclassification of Gerbich phenotypes may occur, but this can be avoided by molecular characterization. This study aimed to characterize the molecular background responsible for rare Gerbich phenotypes in Brazilian population.Materials and MethodsWe selected eight samples from patients with anti‐Ge, six from their relatives and nine samples with normal expression of Gerbich antigens. Serological tests were performed in gel and red blood cells (RBCs) were tested with anti‐Ge2 and anti‐Ge3. Monocyte monolayer assay (MMA) was performed. Molecular investigation was performed with allele‐specific polymerase chain reaction and DNA sequencing.ResultsPatient plasma samples reacted with all commercial RBCs. Patient RBCs showed negative results with anti‐Ge2 and anti‐Ge3. Using MMA two of eight antibodies were clinically significant. Exon 3 was not amplified in any of the patient samples and in two samples from relatives, suggesting the presence of GE*01.‐03/GE*01.‐03. By sequencing, we identified the genetic variability that interferes with the definition of deletion breakpoints, thus two options of genetic structure were suggested to be responsible for the GE:−2,−3,4 phenotype.ConclusionThis study showed for the first time the genetic diversity of GYPC alleles for carriers of Gerbich‐negative phenotypes in a Brazilian population and showed an unexpected prevalence of the GE:−2,−3,4 phenotype. It also demonstrated the importance of using molecular tools to correctly classify Gerbich phenotypes for selection of variants in antigen‐matched transfusions.

Endometriosis is a chronic gynecological disorder defined as the presence of endometrial tissue within extra‐uterine sites. The proliferation of topic and ectopic endometrial tissue was demonstrated in several trials. In endometriotic tissues and cultured endometriotic cells the expression profile of cell cycle and inflammatory proteins are modified, particularly p27 protein – reduced expression – and IL‐1β – increased expression –, both leading to an increased cell proliferation. The therapeutic use of an adenoviral p27 super‐expression vector (Adp27EGFP) in endometriotic cell cultures induces the cells back to normal proliferative levels. In order to improve the therapeutic potential of this gene therapy we combined it with cell therapy using human umbilical cord mesenchymal stem cells (hUCMSC). The levels of IL‐1β were evaluated in the culture medium of normal and endometriotic endometrial cells transfected or not with Adp27EGFP and Adnull, and treated with hUCMSC. Treatment with hUCMSC leads to increased IL‐1β expression reaching endometriotic levels after 14 days of treatment even in the normal endometrial cells. The therapeutic effects of Adp27EGFP were totally inhibited in the endometriotic cells treated with hUCMSC. These results suggest that the mesenchymal stem cells can have a role in the progression of the endometriosis and cannot be used to treat the disease.

Endometriosis seems to be a precursor of epithelial ovarian cancer, especially clear cell and endometrioid adenocarcinomas. The features shared by endometriosis and cancer include the ability to evade apoptosis, the stem cell‐like ability and angiogenic potential. As such characteristics are encoded by the cell's genetic constitution; acquired mutations are responsible for the malignant transformation of endometriosis. So, a number of differences between endometriosis and cancer are found at the molecular level and miRNAs are recognized as key regulators of gene expression. Besides, aberrant expression of these small RNAs has been linked to human disease, including those related to estrogen‐dependent gynecological disorders and cancer. Total RNA of 09 type IV endometriosis patients, as well as, 07 normal endometrioid tissue was extracted, enriched for small RNAs and used for microRNA expression analysis on SOLiD4 sequencer. Sequences were mapped against the miRBase using CLC Genomics Workbench 6.0.1. An average of 5 million reads per sample was mapped on known miRNAs. After mapping and normalization of reads, we used EdgeR to identify differently expressed miRNAs, those with p<0.01 and FDR < 0.05. With this methodology we could obtain differentially expressed miRNAs between the samples which indicates that this approach is feasible for new biomarkers search.