Mesenchymal Stem Cell Microvesicles from Adipose Tissue: Unraveling Their Impact on Primary Ovarian Cancer Cells and Their Therapeutic Opportunities.

Mesenchymal stem cells (MSCs) and their derivatives can be promising tools in oncology including ovarian cancer treatment. This study aimed to determine the effect of HATMSC2-MVs (microvesicles derived from human immortalized mesenchymal stem cells of adipose tissue origin) on the fate and behavior of primary ovarian cancer cells. Human primary ovarian cancer (OvCa) cells were isolated from two sources: post-operative tissue of ovarian cancer and ascitic fluid. The phenotype of cells was characterized using flow cytometry, real-time RT-PCR, and immunofluorescence staining. The effect of HATMSC2-MVs on the biological activity of primary cells was analyzed in 2D (proliferation, migration, and cell survival) and 3D (cell survival) models. We demonstrated that HATMSC2-MVs internalized into primary ovarian cancer cells decrease the metabolic activity and induce the cancer cell death and are leading to decreased migratory activity of tumor cells. The results suggests that the anti-cancer effect of HATMSC2-MVs, with high probability, is contributed by the delivery of molecules that induce cell cycle arrest and apoptosis (p21, tumor suppressor p53, executor caspase 3) and proapoptotic regulators (bad, BIM, Fas, FasL, p27, TRAIL-R1, TRAIL-R2), and their presence has been confirmed by apoptotic protein antibody array. In this study, we demonstrate the ability to inhibit primary OvCa cells growth and apoptosis induction after exposure of OvCa cells on HATMSC2-MVs treatment; however, further studies are needed to clarify their anticancer activities.

Is Maternal Selenium Status Associated with Pregnancy Outcomes in Physiological and Complicated Pregnancy?

Selenium is essential for the synthesis and function of various selenoenzymes, such as glutathione peroxidases, selenoprotein P, and thioredoxin reductase. These enzymes play a critical role in both antioxidant defense and in limiting oxidative damage. Numerous studies have reported associations between serum selenium concentration, obstetric complications and pregnancy outcomes. The aim of this study was to determine whether the dietary intake of selenium, its serum concentration, and the activity of glutathione peroxidase in subsequent trimesters of pregnancy affect the birth condition of newborns. This was assessed based on the APGAR score in the 1st and 5th minute of life, birth weight, body length and head and chest circumference in both physiological and complicated pregnancy courses. Twenty-seven pregnant women, with a mean age of 29.6 ± 4.8 years from the Lower Silesia region of Poland, participated in the study. Fifty-five percent of the study group experienced pregnancy complications. The median reported selenium intake and serum selenium content for Polish pregnant women in the first trimester was 56.30 µg/day and 43.89 µg/L, respectively. These figures changed in the second trimester to 58.31 µg/day and 41.97 µg/L and in the third trimester to 55.60 µg/day and 41.90 µg/L. In the subgroup of pregnant women with a physiological pregnancy course, a weak, positive correlation was observed in the first trimester between Se intake and the length (R = 0.48, p = 0.019) and the birth weight of newborns (R = 0.472, p = 0.022). In the second trimester, a positive correlation was noted with the APGAR score at the 1st (R = 0.680, p = 0.005) and 5th minutes (R = 0.55, p = 0.033), and in the third trimester with the APGAR score at the 1st minute (R = 0.658, p = 0.019). The glutathione peroxidase activity had a strong positive correlation with the APGAR score at the 1st min (R = 0.650, p = 0.008) in the second trimester and with the birth weight of the newborns (R = 0.598, p = 0.039) in the third trimester. No correlation was found between newborns' birth measurements and serum selenium concentration. In the subgroup of pregnant women with complications, a strong, negative correlation was found between Se intake in the second trimester and gestational age (R = -0.618, p = 0.032). In the third trimester, a positive correlation was noted between Se concentration in serum and head circumference (R = 0.587, p = 0.021). The results indicate that maternal selenium status during pregnancy, including dietary intake, serum concentration, and glutathione peroxidase activity, correlates with anthropometric parameters of the newborn, such as birth weight, length, and APGAR score, especially in pregnancies with a physiological course. However, these relationships diminish in importance when pregnancy complications occur.

An unusual coincidence of giant cervical leiomyoma and incidental ovarian granulosa cell tumor: A case report.

RATIONALE: Leiomyomas are the most common benign tumors of smooth muscle origin in women. They are most frequently found in the submucosal tissue of the uterine corpus; however, they also occur in other areas of the uterus, including the cervix. Their size usually varies between 0.5 to 1.0 cm; however, they can reach great dimensions. A strong correlation between the onset and growth of leiomyomas and estrogen levels was observed. Granulosa cell tumor (GCT) is an infrequent sex cord-stromal ovarian neoplasm. Despite their malignancy, GCTs have a good long-term prognosis. In this study, we present a unique case of coincidence of 2 tumors: leiomyoma of rare location (cervix uteri) and extraordinary size (9, 04 cm diameter) with an adult granulosa cell tumor. PATIENT CONCERNS: A 67-year-old Caucasian woman was transported from an emergency ward to a gynecological surgery department due to a massive vaginal hemorrhage. DIAGNOSES: Preliminary examination showed a presence of an enormous uteri cervix tumor. INTERVENTIONS: Initially, the patient underwent physical and ultrasound examinations. To prevent further bleeding, an urgent surgery (hysterectomy) with bilateral salpingo-oophorectomy was performed. OUTCOME: Postoperative histopathological examination revealed a cervical leiomyoma and the incidental occurrence of an adult GCT in the right ovary. LESSONS: This case shares an interesting coincidence between a rare variant of leiomyoma and GCT. The study suggests that the potential reason for this can be estrogen secreted by the GCT, which causes the enormous size of the patient's cervical leiomyoma and the severe vaginal bleeding. Therefore, we advise it is important in abnormal cases to search for other hidden explanations, as in cases of GCT.

Effect of nickel on red blood cell parameters and on serum vitamin B12, folate and homocysteine concentrations during pregnancy with and without anemia.

BACKGROUND: Research to date suggests that nickel affects not only the metabolism of vitamin B12 but also folates and thus may affect hematopoiesis processes. OBJECTIVE: The aim of the study was to examine the relationship of nickel (Ni) status to red blood cell (RBC) parameters and serum vitamin B12, folate and homocysteine concentrations in the course of normal pregnancy and in pregnant women with anemia. METHODS: The study included fifty-three pregnant women recruited to the study from the Lower Silesia region of Poland, 17 % of whom developed anemia. Nickel concentration was determined in urine, whole blood and food samples by atomic absorption spectrometry. At the same time as the food and urine samples were taken, blood was also collected for the determination of RBC parameters and serum vitamin B12, homocysteine and folate concentrations. RESULTS: The median reported Ni intake, and the urinary and whole blood nickel contents for the studied pregnant women for the first trimester were respectively - 162.46 µg/day, 3.98 µg/L and 3.32 µg/L; for the second trimester - 110.48 µg/day, 6.86 µg/L and 1.04 µg/L; and for the third trimester - 132.20 µg/day, 3.41 µg/L and 0.70 µg/L. With regard to Ni concentration in whole blood (p = 0.0204) and in urine (p = 0.0003), the differences in the values for individual trimesters were statistically significant. The whole blood Ni level was significantly higher (9.28 vs 3.62 µg/L, p = 0.0114), while the concentration of homosysteine was significantly lower (4.09 vs 5.04 µmol/L, p = 0.0165) in pregnant women with anemia compared to those without anemia. The whole blood Ni concentration was negatively correlated with almost all RBC parameters in non-anemic pregnant women. CONCLUSIONS: Ni status changes with the development of normal pregnancy, and in the case of anemia, an increase in Ni concentration in whole blood is observed. The demonstrated correlations between the Ni status in pregnant women and RBC parameters as well as serum vitamin B12 and folate concentrations suggest that nickel is associated with the methionine-folate cycle, iron homeostasis and bacterial synthesis of vitamin B12 in humans.

Association of Dietary Inflammatory Index with Serum IL-6, IL-10, and CRP Concentration during Pregnancy.

Background: The mother's diet has a direct impact on fetal development and pregnancy, and can also be important in the course of the body's inflammatory response. An anti-inflammatory diet can be a promising way to counter an excessive inflammatory response in pregnancy. Objective: The aim of the study was to examine the association between the dietary inflammatory index (DII) and the pregnant women's serum interleukin 6 (IL-6) and 10 (IL-10) and C-reactive protein (CRP) concentration in the course of normal and complicated pregnancy. Research Methods and Procedures: The study included 45 Polish pregnant women recruited to the study. The DII, a literature-based dietary index to assess the inflammatory properties of diet, was estimated based on a seven-day 24-h recall and an food frequency questionnaire (FFQ) in each trimester of pregnancy. At the same time as the nutritional interviews, blood samples were collected for the determination of IL-6, IL-10, and CRP concentrations. The studied group was divided into subgroups with normal and complicated pregnancy and depending on the DII median. Results: With the development of pregnancy, the DII score slightly decreased in subsequent trimesters: -1.78 in the first trimester, -2.43 in the second trimester, and -2.71 in the third trimester (p = 0.092). Independent of the trimester of pregnancy and the occurrence of pregnancy complications, the DII score did not affect the differences in the serum concentrations of IL-6, IL-10, and CRP, with the exception of CRP level in the second trimester in women with complicated pregnancy (subgroup with DII < median had a lower CRP level than subgroup with DII > median). In the first and third trimesters, there was a weak but significant positive correlation between the DII score and CRP concentration. During the second trimester, in the group with normal pregnancy and DII below the median, a significant negative correlation between the DII score and the serum IL-6 and IL-10 concentration was noted as well as in the third trimester for IL-6. Conclusion: The anti-inflammatory potential of a pregnant woman's diet increases slightly with pregnancy development; however, its value has no permanent significant association with the level of CRP, IL-6, and IL-10.

Is maternal dietary selenium intake related to antioxidant status and the occurrence of pregnancy complications?

Selenium (Se) is a trace element essential for the appropriate course of vital processes in the human body. It is also a constituent of the active center of glutathione peroxidase and other antioxidant compounds which play an important role in red-ox processes. Associations between lower blood selenium concentration and obstetric complications has been reported in many studies. The aim of this study was to determine the dietary selenium intake and serum selenium content in pregnant Polish women and relate this to antioxidant status as whole blood glutathione peroxidase (GPX) activity, serum uric acid (UA) content and serum total antioxidant status (TAS) and pregnancy complications occurrence. Ninety-four pregnant women at a mean age 30.6 ± 5.4 years from the Lower Silesia region of Poland were recruited to the study, 37% of studied group had pregnancy complications. The mean reported Se intake and serum selenium content for Polish pregnant women was in the first trimester - 53.99 µg/day and 44.36 µg/l, the second trimester - 58.93 µg/day and 43.16 µg/l and the third trimester - 62.89 µg/day and 40.97 µg/l, respectively. Selenium intake below or above recommended value hadn't significant effect on GPX activity, TAS and UA levels. There were no statistical differences in selenium intake, serum selenium content, GPX activity and TAS and UA level between physiological and complicated pregnancy, but a positive correlation between Se intake and serum selenium content was observed during all period of gestation as well as in the second trimester of pregnancy between Se intake and GPX activity in group with physiological pregnancy where selenium intake was below the recommended level. Selenium intake above the recommended level was positively correlated also with serum UA level in first and second trimester of pregnancy. Despite weak, positive correlations in the first two trimesters of pregnancy between selenium supply and GPX activity and UA concentration we concluded that selenium intake does not significantly affect during pregnancy, both: markers of the antioxidant status of pregnant women and the occurrence of pregnancy complications.

Flow cytometric evaluation of intracellular cytokine synthesis in peripheral mononuclear cells of women with endometriosis.

Systemic changes related to cytokine expression levels in women with endometriosis remain a subject of controversy. There are many studies concerning this topic showing differential serum cytokine levels; however, there are limited data presenting cytokine expression at the single-cell level. This study focused on this question by measuring intracellular cytokine staining of activated peripheral CD3+ and CD14+ cells from women with endometriosis (investigative group) compared with those with uterine leiomyoma (control group). Isolated peripheral blood mononuclear cells from women with endometriosis and uterine leiomyoma were stimulated with PMA and ionomycin or with LPS to induce intracellular synthesis of TNF-alpha, IFN-gamma, and IL-8 in subpopulations of CD3+ cells and TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 in CD14+ cells. Comparison of the total groups of patients showed no significant differences in any of the intracellular cytokines investigated in the T cells and monocytes of women with endometriosis compared with controls. When the group of women with endometriosis was divided with regard to severity of disease, a significantly lower percentage of CD3+CD8- lymphocytes stained for IFN-gamma and a significantly higher percentage of CD14+ cells stained for MCP-1 in advanced endometriosis patients compared with the control group were observed. We conclude that peripheral mononuclear cells in women with advanced endometriosis may have differential cytokine synthesis in vitro. These results support the idea that differing immune cell activity measured by intracellular cytokine profiles in women with advanced endometriosis may be more a consequence of the disease than a cause.

Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis.

OBJECTIVE: The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis. STUDY DESIGN: Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells. RESULTS: The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis. CONCLUSIONS: Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.

Studies on selected molecular factors in endometrial cancers.

BACKGROUND: Endometrial carcinomas (EC) differ in etiology, clinical course and prognosis. OBJECTIVES: This multi-center study aimed at a closer recognition of molecular factors linked to heterogeneity of EC by evaluating estrogen and progesterone receptors, proteins dependent on MMR genes, proteins linked to poor prognosis and metastases, and mutations in BRCA1. MATERIAL AND METHODS: Using sections of paraffin-embedded preparations, in 115 patients with EC type I and 31 with EC type II, expression of ERα, ERß1, PR, MLH1, and MSH2 proteins, as well as ARID1A, c-MET and BRCA1, was estimated by immunohistochemistry using specific antibodies. RESULTS: Expression of ERß1 was augmented in EC type II, in poorly differentiated cancers and with growing clinical advancement. An augmented expression of ERα was noted in well-differentiated EC and at lower clinical stage. An increased expression of PR and decreased of MLH1 were detected in type I EC. The expression of ARID1A and c-MET proteins showed no differences between the types of EC, stages of clinical advancement or grading. In 51.6% patients with type II EC, a loss of BRCA1 expression was disclosed; in this group of cancers a decreased expression of ERα was noted. CONCLUSIONS: An augmented expression of ERß1 was linked to type II EC. A higher expression of ERα in EC cancers was associated with a lower histopathological grade. A decreased expression of MLH1 protein was estimated in EC type I. Type II EC may be connected to BRCA1 mutation.

Evaluation of monocyte chemotactic protein-1 levels in peripheral blood of infertile women with endometriosis.

OBJECTIVE: To test whether serum monocyte chemotactic protein-1 (MCP-1) chemokine levels correlate with endometriosis in infertile women. STUDY DESIGN: A group of women with endometriosis (n = 18, infertile) was compared with patients with uterine leiomyoma (n = 16, fertile), unexplained infertility (n = 5, infertile), and healthy women (n = 16, fertile). MCP-1 expression levels were evaluated by ELISA assay. The data obtained were statistically analyzed using the Mann-Whitney test. P-Values <0.05 were considered as significant. RESULTS: MCP-1 concentrations (median; range of values) in serum were as follows: women with endometriosis (221; 101-635 pg/ml), women with unexplained infertility (167, 114-234 pg/ml), women with uterine leiomyoma (137; 88-200 pg/ml), and healthy donors (123; 98-194 pg/ml). Significant differences were observed in the women with endometriosis compared with those with uterine leiomyoma (p = 0.02) and healthy donors (p = 0.002). Among the women with endometriosis, the level of significance in MCP-1 level at rAFS stages III-IV was higher than that at rAFS stages I-II compared with healthy donors and women with leiomyoma (p = 0.002 and p = 0.02, respectively). CONCLUSIONS: These data show that an increased level of MCP-1 can characterize infertile women with endometriosis. However, further studies are needed to be able to determine whether increased MCP-1 chemokine expression can be related to infertility or is a result of endometriosis progress.

Ectopic and eutopic stromal endometriotic cells have a damaged ceramide signaling pathway to apoptosis.

OBJECTIVE: To investigate whether sphingosine analogues, which activate the ceramide signaling pathway to apoptosis, can cause the death of ectopic (EEC) and eutopic stromal endometriotic cells (EEU), as well as healthy eutopic stromal endometrial cells (HEU). DESIGN: The EEC, EEU, and HEU isolated from fertile and infertile women with endometriosis were cultured for 48 hours in RPMI medium with 10% fetal calf serum (FCS) and with 2.5-10 microM sphingosine analogues. SETTING: A clinic for the treatment of endometriosis and basic research laboratories. PATIENT(S): Nineteen women with follicular cyst and 16 women with endometriosis. MAIN OUTCOME MEASURE(S): The percentage of proliferating cells was determined by 93-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis and cell cycle were detected by fluorescence-activated cell sorter (FACS) Calibur flow cytometer. RESULT(S): The viability of EEC after exposure to 10 microM sphingosine analogues was 59.5% +/- 9.7% for D-sphingosine and 77.65 +/- 9.7% for DL-erythro-sphingosine, the viability of EEU was 69.2% +/- 14.2% and 42.0% +/- 15.5%, whereas the viability of comparative HEU was 9.0% +/- 4.8% and 18.8% +/- 8.3%, respectively. The differences were significant using the Mann-Whitney test. The apoptotic level of the cells treated with 10 microM sphingosine analogues for comparative HEU was 42.8% +/- 7.5% for D-sphingosine and 42.5% +/- 10.5% for DL-erythro-sphingosine, whereas for EEC this was 16.7% +/- 5.5% for D-sphingosine and 14.1% +/- 4.4% for DL-erythro-sphingosine and for EEU this was 14.3% +/- 4.7% and 22.9% +/- 8.9%, respectively. CONCLUSION(S): Ectopic and eutopic stromal endometrial cells from women with endometriosis have a damaged ceramide-downstream pathway to apoptosis.

The influence of extracellular matrix proteins on T-cell proliferation and apoptosis in women with endometriosis or uterine leiomyoma.

PROBLEM: Interactions between the extracellular matrix (ECM) and peripheral blood T cells in women with endometriosis and leiomyoma are hardly unknown. We have investigated the influence of two major ECM components, collagen IV (C-IV) and fibronectin (Fn), on T-cell proliferation and apoptosis in women with endometriosis and uterine leiomyoma. beta1 integrin expression, responsible for interactions with ECM proteins, was also studied. METHOD OF STUDY: Peripheral blood lymphocytes were obtained from 53 women (17 with uterine leiomyomas, 18 with endometriosis, and 18 from healthy donors). T cells were exposed to ECM proteins co-immobilized with monoclonal antibody anti-CD3 for 72 hr. Apoptosis and S phase of the cell cycle of the T cells were studied by DNA analysis using flow cytometry. The proliferation of T cells was evaluated by MTT assay. The percentage of CD3+ cells expressing CD29 (beta1 integrin chain) was evaluated by double-color flow cytometry. Results were analyzed statistically using the Mann-Whitney test. RESULTS AND CONCLUSIONS: (1) A general increase in the percentage of T cells in S phase could be seen in women with endometriosis and uterine leiomyoma in all culture conditions what may suggest general activation of T cells. (2) A significant increase in the percentage of cells in S phase was shown only in the case of T cells exposed to anti-CD3 + C-IV in both women with uterine leiomyoma and endometriosis. (3) However, no apoptotic cells were observed. (4) T cells from patients with uterine leiomyoma exhibited significantly increased level of proliferation after culture with anti-CD3 + C-IV. (5) More T cells expressed beta1 integrin in women with endometriosis or uterine leiomyoma than in healthy donors. Our data may suggest that increased beta1 integrin expression may enhance T-cell-ECM interactions, which may be responsible for the increased proliferation of T cells but not for apoptosis. Therefore, it is possible that interactions of T cells with ECM proteins, especially with C-IV, may contribute to the pathogenesis of endometriosis and uterine leiomyoma.