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Endometriosis, the presence of ectopic endometrium, has an unclear etiology and is commonly associated with endocrine, genetic, and immunological imbalance. This study determined whether immunomodulation by the RESAN vaccine could alter the potentially pathogenic gene expression profiles in the cells of the eutopic endometrium in an animal model of endometriosis. Preventing these changes could inhibit the early development of the illness and support the success of surgical treatment. Wistar rats were divided into three groups: prophylaxis (vaccinated before ectopic endometrium implantation, n = 23), therapeutic (vaccinated at the time of the ectopic excision, n = 23) and control (n = 10). During the first laparotomy, autotransplantation of the endometrium to the peritoneum was performed in the prophylaxis and therapeutic groups. The second laparotomy was carried out three months later in all groups to examine endometriotic foci and adhesions. Suspected endometriosis foci were removed. Three months later, the third laparotomy was performed in all animals, followed by suspected foci excision. Fragments of the eutopic endometrium were collected from all animals during the first and third laparotomies. All samples were analysed by real-time PCR to assess the expression of Bcl2, Bax, Bax/Bcl2 ratio, Mki67, and Tert genes. Endometrial foci were found in abdominal peritoneum at the second laparotomy in 1 animal in the prophylaxis group, compared to 16 animals in the therapeutic group. The prophylaxis group showed a high expression of Bax while the therapeutic group showed high expression of Bax, Tert and Mki67 genes. Additional analysis revealed that throughout the six months of the experiment, the expression of the Bax, Tert, and Mki67 genes decreased significantly in the prophylaxis group, Mki67 gene expression decreased in the therapeutic group, and Tert, Mki67, and Bcl2 gene expression decreased in the control group. The results indicate that immunomodulation affects the balance between apoptosis and proliferation in the eutopic endometrium and may prevent the onset and recurrence of endometriosis. [ABSTRACT FROM AUTHOR]

Abstract Background Metastasis is a common feature of many advanced stage cancers and metastatic spread is thought to be responsible for cancer progression. Most cancer cells are localized in the primary tumor and only a small population of circulating tumor cells (CTC) has metastatic potential. CTC amount reflects the aggressiveness of tumors, therefore their detection can be used to determine the prognosis and treatment of cancer patients. The aim of this study was to evaluate human chorionic gonadotropin beta subunit (CGB) and gonadoliberin type 1 (GNRH1) expression as markers of tumor cells circulating in peripheral blood of gynecological cancer patients, indicating the metastatic spread of tumor. Methods CGB and GNRH1 expression level in tumor tissue and blood of cancer patients was assessed by real-time RT-PCR. The data was analyzed using the Mann-Whitney U and Spearman tests. In order to distinguish populations with homogeneous genes' expression the maximal likelihood method for one- and multiplied normal distribution was used. Result Real time RT-PCR results revealed CGB and GNRH1 genes activity in both tumor tissue and blood of gynecological cancers patients. While the expression of both genes characterized all examined tumor tissues, in case of blood analysis, the transcripts of GNRH1 were found in all cancer patients while CGB were present in 93% of patients. CGB and GNRH1 activity was detected also in control group, which consisted of tissue lacking cancerous changes and blood of healthy volunteers. The log-transformation of raw data fitted to multiplied normal distribution model showed that CGB and GNRH1 expression is heterogeneous and more than one population can be distinguished within defined groups. Based on CGB gene activity a critical value indicating the presence of cancer cells in studied blood was distinguished. In case of GNRH1 this value was not established since the results of the gene expression in blood of cancer patients and healthy volunteers were overlapping. However one subpopulation consists of cancer patient with much higher GNRH1 expression than in control group was found. Conclusions Assessment of CGB and GNRH1 expression level in cancer patients' blood may be useful for indicating metastatic spread of tumor cells.

Abstract Background Corydalis cava Schweigg. & Koerte, the plant of numerous pharmacological activities, together with the studied earlier by our group Chelidonium majus L. (Greater Celandine), belong to the family Papaveraceae. The plant grows in Central and South Europe and produces the sizeable subterraneous tubers, empty inside, which are extremely resistant to various pathogen attacks. The Corydalis sp. tubers are a rich source of many biologically active substances, with the extensive use in European and Asian folk medicine. They have analgetic, sedating, narcotic, anti-inflammatory, anti-allergic and anti-tumour activities. On the other hand, there is no information about possible biological activities of proteins contained in Corydalis cava tubers. Methods Nucleolytic proteins were isolated from the tubers of C. cava by separation on a heparin column and tested for DNase activity. Protein fractions showing nucleolytic activity were tested for cytotoxic activity in human cervical carcinoma HeLa cells. Cultures of HeLa cells were conducted in the presence of three protein concentrations: 42, 83 and 167 ng/ml during 48 h. Viability of cell cultures was appraised using XTT colorimetric test. Protein fractions were separated and protein bands were excised and sent for identification by mass spectrometry (LC-ESI-MS/MS). Results The studied protein fractions showed an inhibiting effect on mitochondrial activity of HeLa cells, depending on the administered dose of proteins. The most pronounced effect was obtained with the highest concentration of the protein (167 ng/ml) - 43.45 ± 3% mitochondrial activity of HeLa cells were inhibited. Mass spectrometry results for the proteins of applied fractions showed that they contained plant defense- and pathogenesis-related (PR) proteins. Conclusions The cytotoxic effect of studied proteins toward HeLa cell line cells has been evident and dependent on increasing dose of the protein. The present study, most probably, represents the first investigations on the effect of purified PR proteins from tuber extracts of a pharmacologically active plant on cell lines.

Aim. To assess the role of the Th1 and Th2 cellular response in the etiology of endometriosis observed in a rat model, with the use of the RESAN immunomodulator. Materials and Methods. A comparative analysis of cytokines in blood serum typical of Th1 (TNF-α and INF-γ) and Th2 (IL-4, IL-6, IL-10) cell response in groups of rats, in which RESAN preparation was used as prophylaxis (Gr. I) or treatment (Gr. II) of endometriosis. Results. The results indicated an increase in the level of cytokines in blood serum typical of Th2 cell response by comparing the second and third stages of the experiment in the second group of rats and a decrease in IL-4 and IL-10 between III and IV stages. There was a significant difference in cytokine levels during the third stage of the experiment by comparing I and II groups of rats. In the III group of rats, levels of IL-10 significantly increased between the II and III stages of the experiment. Conclusion. RESAN preparation shows Th2 cell response, inhibiting the development of endometriosis in a rat model. Due to successful prophylactic action, one may speculate that RESAN vaccine may be effective as a complementary treatment after surgical excision. [ABSTRACT FROM AUTHOR]

Objectives: The presence of the endometrium outside the uterine cavity affects about 10% of women of childbearing age. Studies of the progression of endometriosis to cancer have been supported by numerous evidences of gene expression or gene defect caused by oxidative stress and inflammation. We decided to check the expression of selected factors responsible for the proliferation, as in the stages of neoplasia., Material and Methods: A group of 80 women with ovary localization of endometriosis was qualified for research. The control group was 90 patients with ovarian simplex or follicular cysts. The DNA isolation, immunohistochemical analysis of IGF 1, IGF-R, TSG 101, and LSF expressions with a quantitative scoring of slides and electron microscopy was performed., Results: The IGF-1-immunopositive cells in the reference group were in statistically significantly higher number compared to the cells forming the foci of endometriosis (p = 0.0282). However, the number of IGF-R-immunopositive cells was comparable to the endometriosis (p = 0.1264). In the control group, the number of LSF-immunopositive cells was statistically significantly higher in comparison to endometriosis foci (p = 0.000001), but the number of TSG 101-immunositive cells was comparable to endometriosis foci (p = 0.3834). A weak negative correlation between the number of cells expressing the TSG 101 factor and the IGF-1 receptor was found in the endometriosis group (r = -0.26, p = 0.0196). The analysis of CA single nucleotide polymorphism in the DNA isolated from both groups showed a comparable incidence of MSS and MSI-L genotypes (chi2 p = 0,9160)., Conclusions: How these factors affect the development of endometriosis and whether they could be helpful in the diagnosis requires further research.

This work reports for the first time on a new class of flexible polyurethane foam hybrids (PUFs) synthesized with the use of less toxic aliphatic hexamethylene diisocyanate (HDI), which have been chemically modified by POSS moieties. The flexible polyurethane foam hybrids (PUFs) chemically modified by functionalized polyhedral oligomeric silsesquioxanes: octa(3-hydroxy-3-methylbutyldimethylsiloxy)POSS (OCTA-POSS) and 1,2-propanediolizo-butylPOSS (PHI-POSS), was obtained. The resulting foams, which contain 0 to 15 wt % POSS, were characterized in terms of their structure, morphology, density and compressive strength. The FT-IR results indicate the chemical incorporation of both OCTAPOSS and PHIPOSS into the polyurethane matrix. SEM-EDS analysis showed that both OCTAPOSS and PHIPOSS nanoparticles are distributed homogeneously in the foam structure; at 15 wt % load PHIPOSS characteristic "crosses" are formed. With the increase of PHIPOSS content in the matrix, the formation of agglomerates is observed, as revealed by WAXD spectra. The introduction of POSS compounds reduces the porosity of the polyurethane, with the number of pores increasing as the amount of modifier increases. Mechanical tests - compressive strength - show that the hardness of modified materials (5 wt % POSS) increases compared to the reference material. An incubation was carried out in a simulated physiological fluid (SBF) to pre-assess the bioactivity of the materials obtained. The obtained results confirmed the formation of a hydroxyapatite layer on the PUF-POSS surface. Cytotoxicity, cell cycle and apoptosis of osteoblast cells and fibroblasts were assessed in the presence of the PUF-POSS materials. Test materials have a cytotoxic effect on both established cell lines. PUF-PHIPOSS samples showed better biocompatibility than reference and PUF-OCTAPOSS samples, as they caused lower mortality of the examined cells., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Cervical microbial communities serve a crucial role in the persistence and development of oncogenic human papilloma virus (HPV) infections. In the present study, the authors hypothesised that disturbed heterogeneity of microbial flora was associated with HPV-induced carcinogenesis. Swabs of the cervical microbiota were collected from 250 women and the 16S ribosomal DNA was sequenced using a high throughput assay. The swabs of cervical microbiota were grouped according to the community state types (CSTs) as follows: Healthy cervical swabs; swabs taken from low-grade squamous intra-epithelial lesions (LSIL) and swabs taken from high-grade squamous intra-epithelial lesions (HSIL). Analysis of the bacterial classes revealed that the CST cervical swabs of the volunteers were characterised by Lactobacillus crispatus, Lactobacillus iners and Lactobacillus taiwanensis , however, Gardnerella vaginalis and Lactobacillus acidophilus were absent. In the CST of patients with LSIL the predominant type of bacteria was Lactobacillus acidophilus and Lactobacillus iners , however Lactobacillus crispatus was not detected. Swabs from CST women diagnosed with HSIL exhibited abundant Gardnerella vaginalis and Lactobacillus acidophilus , however, lacked Lactobacillus taiwanensis, Lactobacillus iners and Lactobacillus crispatus . The abundance of Lactobacillus acidophilus in swabs from the healthy women was compared with the swabs from the women with LSIL. The results of the present study indicated that the development of HPV-induced cancer is associated with a high diversity of vaginal microbiota, which is involved in the control of viral persistence, and is therefore indicative of disease prognosis.

Endometrial carcinoma (EC) is the most common type of gynecological malignancy. Studies have demonstrated that the insulin growth factor (IGF) pathway is implicated in the development of endometrial tumors and that the serum levels of IGF‑1 are affected by estrogen. Most EC cells with high microsatellite instability (MSI‑H) accumulate mutations at a microsatellite sequence in the IGF‑1 gene. The present study investigated the CA repeat polymorphism in the P1 promoter region of the IGF‑1 gene among Caucasian females with endometrial hyperplasia, EC and healthy control subjects, whose blood serum and surgical tissue specimens were analyzed. Differences or correlations between the analyzed parameters [serum levels of IGF-1 and IGF binding protein (IGFBP)‑1 and IGFBP‑3 as well as estrogens among the polymorphisms] were verified using the χ2, Mann-Whitney U, Kruskal-Wallis or Spearman's rank correlation tests. A PCR amplification and DNA sequencing analysis was used for identification of (CA)n repeats in the P1 region of IGF‑1. ELISA was used to determine the blood serum levels of IGF‑1, IGFBP‑1, IGFBP‑3 and estrogens. Furthermore, IGF-1 was assessed in endometrial tissues by immunohistochemical analysis. The present study indicated no statistically significant differences between serum levels of IGF‑1, IGFBP‑1, IGFBP‑3 and estrone, estriol and estradiol in the control and study groups. A significant correlation was identified between the IGF-1 levels and estrone levels in the MSI-H polymorphism (r=-0.41, P=0.012) as well as a highly negative correlation between IGF-1 levels and the estradiol levels in the MSI-H polymorphism (r=-0.6, P=0.002). Genotypes without the 19 CA allele were predominantly found in EC. Furthermore, statistical analysis indicated that the number of IGF-1-expressing cells was significantly elevated in MSI-H type 18-20 (P=0.0072), MSI-L type 19-20 (P=0.025) and microsatellite-stable MSS type 19-19 (P=0.024) compared with those in the MSI-H 20-20 genotype. The present study suggested that it is rather likely that the polymorphisms in the IGF-1 promoter are associated with EC in Caucasian females with regard to its development. In the present study, polymorphisms of the IGF-1 promoter may have been introduced during the genesis of EC and contributed to it by leading to aberrant expression of IGF-1.

High oncogenic risk human papillomaviruses (HPVs) are closely associated with cancer of the cervix. However, HPV infection alone may not be sufficient to cause cervical cancer, and other factors or cofactors may have a cumulative effect on the risk of progression from cervical HPV infection to cancer. The present study investigates the cytosine‑adenine (CA) repeat polymorphism in the P1 promoter region of the insulin‑like growth factor‑1 (IGF‑1) gene among cervical precancerous and cancer patients and healthy control females. The association between these polymorphisms, tissue and blood serum levels of IGF‑1, and cervical cancer risk and progression is evaluated. The material for analysis consisted of blood cells and postoperative tissues from patients diagnosed with low‑grade squamous intraepithelial lesions (L‑SILs), high‑grade squamous intraepithelial lesions (H‑SILs) and invasive cervical cancer (ICC). A polymerase chain reaction amplification and the sequencing of DNA were used for the identification of (CA)n repeats in the IGF‑1 P1 region and detection of HPV DNA. The blood serum concentration of IGF was determined by enzyme‑linked immunosorbent assay. The identification of the IGF‑1 protein in the cervical tissues was performed by immunohistochemical analysis. The range of the length of the CA repeats in the study DNA was 11 to 21. However, the most common allele length and genotype in the control and study patients from serum and tissues was 19 CA repeats and a homozygous genotype of CA19/19. Statistically significant differences in the concentration of IGF‑1 in the blood serum were observed between H‑SILs and controls, only (p=0.047). However, the concentration of IGF‑1 in the group of females with CA19/19, CA19<19 and CA19>19 was significantly higher in the group of patients with H‑SIL (P=0.041) and ICC (P=0.048) in comparison with the control group. An association was detected between CA repeat length <19 and/or >19, IGF concentration in blood serum and tissues and the development of cervical cancer.

Current techniques of in vitro cell cultures are able to mimic the in vivo environment only to a limited extent, as they enable cells to grow only in two dimensions. Therefore cell culture approaches should rely on scaffolds that provide support comparable to the extracellular matrix. Here we demonstrate the advantages of novel nanostructured three-dimensional grids fabricated using electro-spinning technique, as scaffolds for cultures of neoplastic cells. The results of the study show that the fibers allow for a dynamic growth of HeLa cells, which form multi-layer structures of symmetrical and spherical character. This indicates that the applied scaffolds are nontoxic and allow proper flow of oxygen, nutrients, and growth factors. In addition, grids have been proven to be useful in in situ examination of cells ultrastructure.

Objectives: In our study, with the use of GH3 cells line we decided to examine 1) what is the relation between the dose of bromocriptine and the development of apoptosis in GH3 cells 2) whether the induction of apoptosis is accompanied by alterations in bcl-2 and p53 content and 3) whether dibutyryl-cAMP or phorbol esters affect the initiation of apoptosis in GH3 cells., Results: The current study demonstrated the absence of alterations in GH3 cells incubated for 24 h with bromocriptine at the concentrations of up to 15 micromol/l. Apoptotic and necrotic changes were observed after 48 h incubation with bromocriptine at the concentrations of 25 micromol/l. The ratio of necrotic to apoptotic cells increased at 40 micromol/l of bromocriptine concentration. An inhibitory effect of bromocriptine on cell proliferation was also observed. Phorbol-12-myristate-13-acetate (PMA), at concentrations ranging between 25 ng to 200 ng/ml, reduced the amount of apoptotic cells., Conclusions: Application of dibutyryl-cAMP at the concentration of 1 to 8 mmol/l resulted in an inhibition of apoptosis, followed by an increase in the number of cultured cells. Ultrastructural studies showed evident apoptotic lesions in the cells.