Assessment of breast cancer progression and metastasis during a hypercoagulable state induced by silencing of antithrombin in a xenograft mouse model.

Local coagulation activation has been shown to impact both primary tumor growth and metastasis in mice. It is well known that components of the blood clotting cascade such as tissue factor and thrombin play a role in tumor progression by activating cellular receptors and local formation of fibrin. However, whether venous thromboembolism (VTE) or a hypercoagulable state has a direct impact on cancer progression is unknown. Here we have combined an orthotopic murine breast cancer model, using female Nod-SCID mice, with siRNA-mediated silencing of antithrombin (siAT) leading to the induction of a systemic hypercoagulable state. We show that, compared to control siRNA-treated (not experiencing a hypercoagulable state) tumor-bearing mice, siAT treated tumor-bearing mice do not show enhanced tumor growth nor enhanced metastasis. We conclude that, in this murine model for hypercoagulability, induction of a hypercoagulable state does not contribute to breast cancer progression.

Genes associated with venous thromboembolism in colorectal cancer patients.

Essentials The underlying pathophysiological mechanisms behind cancer-associated thrombosis are unknown. We compared expression profiles in tumor cells from patients with and without thrombosis. Tumors from patients with thrombosis showed significant differential gene expression profiles. Patients with thrombosis had a proinflammatory status and increased fibrin levels in the tumor. SUMMARY: Background Venous thromboembolism (VTE) is a frequent complication in patients with cancer, and is associated with significant morbidity and mortality. However, the mechanisms behind cancer-associated thrombosis are still incompletely understood. Objectives To identify novel genes that are associated with VTE in patients with colorectal cancer (CRC). Methods Twelve CRC patients with VTE were age-matched and sex-matched to 12 CRC patients without VTE. Tumor cells were isolated from surgical samples with laser capture microdissection approaches, and mRNA profiles were measured with next-generation RNA sequencing. Results This approach led to the identification of new genes and pathways that might contribute to VTE in CRC patients. Application of ingenuity pathway analysis indicated significant links with inflammation, the methionine degradation pathway, and increased platelet function, which are all key processes in thrombus formation. Tumor samples of patients with VTE had a proinflammatory status and contained higher levels of fibrin and fibrin degradation products than samples of those without VTE. Conclusion This case-control study provides a proof-of-principle that tumor gene expression can discriminate between cancer patients with low and high risks of VTE. These findings may help to further unravel the pathogenesis of cancer-related VTE. The identified genes could potentially be used as candidate biomarkers to select high-risk CRC patients for thromboprophylaxis.

Interplay between alternatively spliced Tissue Factor and full length Tissue Factor in modulating coagulant activity of endothelial cells.

BACKGROUND: Full length Tissue factor (flTF) is a key player in hemostasis and also likely contributes to venous thromboembolism (VTE), the third most common cardiovascular disease. flTF and its minimally coagulant isoform, alternatively spliced TF (asTF), have been detected in thrombi, suggesting participation of both isoforms in thrombogenesis, but data on participation of asTF in hemostasis is lacking. Therefore, we assessed the role of asTF in flTF cofactor activity modulation, using a co-expression system. OBJECTIVE: To investigate the interplay between flTF and asTF in hemostasis on endothelial cell surface. METHODS: Immortalized endothelial (ECRF) cells were adenovirally transduced to express asTF and flTF, after which flTF cofactor activity was measured on cells and microvesicles (MVs). To study co-localization of flTF/asTF proteins, confocal microscopy was performed. Finally, intracellular distribution of flTF was studied in the presence or absence of heightened asTF levels. RESULTS: Levels of flTF antigen and cofactor activity were not affected by asTF co-expression. asTF and flTF were found to localize in distinct subcellular compartments. Only upon heightened overexpression of asTF, lower flTF protein levels and cofactor activity were observed. Heightened asTF levels also induced a shift of flTF from non-raft to lipid raft plasma membrane fractions, and triggered the expression of ER stress marker BiP. Proteasome inhibition resulted in increased asTF - but not flTF - protein expression. CONCLUSION: At moderate levels, asTF appears to have negligible impact on flTF cofactor activity on endothelial cells and MVs; however, at supra-physiological levels, asTF is able to reduce the levels of flTF protein and cofactor activity.

The evaluation of hormonal and psychological parameters that affect bone mineral density in postmenopausal women.

OBJECTIVE: In this study we aimed to investigate the relationships between serum levels of DHEAS, reproductive hormones and low bone mineral density (BMD) in postmenopausal women. We also examined the relationship between psychological status of patients and their BMD results. PATIENTS AND METHODS: This study included postmenopausal female patients. BMD measurements were performed with dual-energy X-ray absorptiometry (DEXA). Psychological assessments of all cases were performed using the Hamilton Anxiety and Hamilton Depression scales. All patients provided fasting venous blood samples in order to determine serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and DHEAS. RESULTS: There were 33 cases (45.2%) with normal BMD levels and 40 cases (54.8%) with abnormal BMD levels (osteopenia and osteoporosis). DHEAS levels did not show any statistically significant difference according to BMD results (p = 0.431). The Hamilton Anxiety and Depression scores also did not show statistically significant differences in accordance with the BMD results (p = 0.889 and p = 0.706, respectively). CONCLUSIONS: According to our results, anxiety, depression and circulating DHEAS levels are not significantly associated with low levels of BMD, particularly at osteopenic levels. So these parameters are not useful for clinical practice in patients with low BMD in the middle-aged postmenopausal women.

Adrenal gland volume assessed by magnetic resonance imaging in women with polycystic ovary syndrome.

PURPOSE: To determine adrenal gland volume (AGV) in women with polycystic ovary syndrome (PCOS) by comparison with healthy control subjects and to investigate the relationship between AGV and hormonal status. PATIENTS AND METHODS: AGV was measured on transverse sections of T1-weighted MRI imaging data in 27 PCOS patients and 40 age-matched control subjects for this prospectively designed study. A comparative analysis of AGV in PCOS and controls was performed and possible correlations between AGV and hormonal parameters were evaluated. RESULTS: PCOS patients had significantly larger AGV compared to controls ((11.7±4.4 cm(3), 7.2±1.9 cm(3), respectively, P<0.001). A significant positive correlation was found between total AGV and dehydroepiandrosterone sulfate, 17-OH progesterone, and total and free testosterone levels in the PCOS group (r=+0.51, +0.48, +0.43, +0.62, respectively; P values<0.05). In addition, AGV was significantly negatively correlated with LH and LH/FSH ratio in the PCOS group (r= -0.55, P=0.02; r=-0.51, P=0.01, respectively). CONCLUSIONS: PCOS patients have significantly increased AGV as well as a positive correlation of AGV and androgens. We conclude that the assessment of AGV with MRI shows a significant correlation with the androgenic activity of the gland, and that hypertrophy of the adrenal gland may be involved in the pathogenesis of PCOS.

Alternatively spliced tissue factor synergizes with the estrogen receptor pathway in promoting breast cancer progression.

BACKGROUND: Procoagulant full-length tissue factor (flTF) and its minimally coagulant alternatively spliced isoform (asTF), promote breast cancer (BrCa) progression via different mechanisms. We previously showed that flTF and asTF are expressed by BrCa cells, resulting in autoregulation in a cancer milieu. BrCa cells often express hormone receptors such as the estrogen receptor (ER), leading to the formation of hormone-regulated cell populations. OBJECTIVE: To investigate whether TF isoform-specific and ER-dependent pathways interact in BrCa. METHODS: Tissue factor isoform-regulated gene sets were assessed using ingenuity pathway analysis. Tissues from a cohort of BrCa patients were divided into ER-positive and ER-negative groups. Associations between TF isoform levels and tumor characteristics were analyzed in these groups. BrCa cells expressing TF isoforms were assessed for proliferation, migration and in vivo growth in the presence or absence of estradiol. RESULTS: Ingenuity pathway analysis pointed to similarities between ER- and TF-induced gene expression profiles. In BrCa tissue specimens, asTF expression was associated with grade and stage in ER-positive but not in ER-negative tumors. flTF was only associated with grade in ER-positive tumors. In MCF-7 cells, asTF accelerated proliferation in the presence of estradiol in a ß1 integrin-dependent manner. No synergy between asTF and the ER pathway was observed in a migration assay. Estradiol accelerated the growth of asTF-expressing tumors but not control tumors in vivo in an orthotopic setting. CONCLUSION: Tissue factor isoform and estrogen signaling share downstream targets in BrCa; the concomitant presence of asTF and estrogen signaling is required to promote BrCa cell proliferation.

In contrast to leptin, serum concentrations of ghrelin are not related to premenstrual syndrome.

OBJECTIVE: Premenstrual syndrome (PMS) is a disorder related to mood and appetite changes during the premenstrual phase. Unfortunately, the understanding of the pathophysiology of PMS is quite poor. Though, ghrelin and leptin play important roles in the control of food intake. The aim of this study was to evaluate leptin and ghrelin serum concentrations in PMS patients. PATIENTS AND METHODS: Forty-five PMS patients diagnosed according to ICD-10 diagnostic criteria and 45 healthy women as a control group, were included in the study. These groups were matched for age, body mass index and duration of menstrual cycle. Symptoms of the patients were evaluated using "Menstrual Distress Questionnaires". Serum leptin and ghrelin serum concentrations were measured using ELISA in the postmenstrual phase (5-9 days) and 2-3 days before menstruation. Mann-Whitney U test, independent sample t-test and Wilcoxon test were used for statistical analyses. RESULTS: In the PMS group, there was no difference in the serum concentrations of ghrelin; however, leptin serum concentrations were 31.05 (± 14.16) and 16.42 (± 15.81) ng/ml during the premenstrual and postmenstrual periods, respectively (p < 0.05). Ghrelin serum concentrations in the premenstrual period were 6.9 (± 9.3) ng/ml in the PMS group and 8.8 (± 9.3) ng/ml in the control group, but this difference was not statistically significant (p = 0.79). CONCLUSIONS: Ghrelin serum concentrations were not associated with PMS, while leptin serum concentrations were found to be higher in the premenstrual period in PMS patients. Though, these two hormones work antagonistically to control the food intake and body weight, we suggest that this function is not relevant to PMS.

Can first trimester placental protein-13 and pregnancy-associated plasma protein-A predict pre-eclampsia in Turkish women?

The aim of study was to evaluate placental protein-13 (PP-13) and pregnancy-associated plasma protein-A (PAPP-A) in first trimester maternal serum, for predicting pre-eclampsia. A prospective case-control study included 30 pre-eclampsia patients and 90 control pregnant women. Pre-eclampsia patients were divided into two subgroups: early- and late-onset (9 vs 21), and PP-13 and PAPP-A levels were compared between the groups and the comparison of risks for pre-eclampsia were calculated. Results showed that there was a significant inverse correlation between PAPP-A and late pre-eclampsia (p = 0.003), with a cut-off value of 0.805 (ROC analysis area under curve = 0.751). There was a significant reverse correlation between PAPP-A and early pre-eclampsia (p = 0.02). There was no significant relationship between PP-13 and early pre-eclampsia, nor with late pre-eclampsia (p = 0.7, p = 0.6, respectively). It was concluded that neither of these markers can serve as a sufficient and reliable screening test of pre-eclampsia because of inadequate sensitivity in the Turkish pregnant population.

Maternal serum pregnancy-associated plasma protein-A levels in hyperemesis gravidarum: a prospective case control study.

PURPOSE: To investigate if maternal serum pregnancy-associated plasma protein-A levels are affected in hyperemesis gravidarum (HG). MATERIALS AND METHODS: A prospective case control study was conducted in 169 HG cases who had one or more antepartum hospitilization for HG. The control pregnancies were 132 and were selected randomly among all women who had first trimester prenatal screening in antenatal outpatient clinic between 2011 and 2012. RESULTS: Maternal serum pregnancy-associated plasma protein-A levels were significantly higher in hyperemesis gravidarum group compared with control group (p = 0.002 p < 0.05 95% CI). Power analysis of independent sample t-test, two-sided, for pregnancy-associated plasma protein-A was 0.88. Maternal serum free beta-human chorionic gonadotropin values were not different between two groups (p > 0.05). CONCLUSION: Increased pregnancy-associated plasma protein-A levels associated with HG, even after excluding potential cofounders.

Role of omentectomy and appendectomy in surgical staging of endometrioid endometrial cancer.

PURPOSE: The purpose of this study was to determine whether it was necessary to add omentectomy and appendectomy to the surgical staging of endometrioid endometrial cancer. MATERIALS AND METHODS: Records were reviewed from June 2005 to June 2009 for endometrioid endometrial cancer patients who underwent total abdominal hysterectomy, bilateral salpingo-oophorectomy, pelvic and para-aortic lymphadenectomy, infracolic omentectomy and appendectomy. RESULTS: In total, 186 patients were included in the analysis. Disease was limited to uterus in 93% of patients and 87% of patients had Stage I disease. There was only one omental metastasis and no appendix metastasis in all stages. CONCLUSION: Routine omentectomy and appendectomy are unnecessary in surgical staging of endometrioid endometrial cancer unless there is suspicion of gross metastases during intraoperative examination.

Comparative evaluation of the nutrient and COD removal performances of various sequentially operated biofilm reactors.

In this paper, the performances of Fixed Bed (FXB), Fluidized Bed (FLB), and Hybrid (HYB, i.e. fluidized bed followed by fixed) operational conditions of biofilm reactors were investigated. The COD (chemical oxygen demand), TN (total nitrogen) and TP (total phosphorus) were taken as performance-indicator parameters. For that purpose, a pilot experimental setup allowing FXB, FLB and HYB operations in a single reactor was run under varying COD (between 500 to 2000 mg COD l(-1)), TN (between 25 to 100 mg l(-1)) and TP (between 5 to 20 mg l(-1)) influent concentrations. The system was operated as sequentially batch (SBR) and a filtering process was added at the end of each operational phase in order to achieve liquid-solid separation. Results indicated that FXB and FLB are two upper and lower cases, and HYB plays a role between them and respond as a best alternative. For example, for the lower influent concentrations (around 500 mg l(-1)), 84% of COD was removed to final averages of 78 +/- 15.4 mg l(-1); simultaneously, TN reduced to 18.55 +/- 3.48 mg l(-1) (corresponding to 31% TN removal) and 60% of TP was treated to final averages of 1.92 +/- 0.2 mg l(-1) in the HYB operation. When the influent concentrations were increased to 1000 mg l(-1) , COD removal efficiency of the HYB reactor was reduced to 73% (COD reduced from 1005.3 +/- 6.13 mg l(-1) to 270.57 +/- 13.18 mg l(-1)) and, due to incomplete organic matter degradation and oxygen deficiency, 40% of TN was removed to final averages of 34.03 +/- 5.04 mg l(-1) and 56% of TP treated to final averages of 3.98 +/- 0.28 mg l(-1). Performance of the HYB reactor was decreased when influent concentration increased to 2000 mg COD l(-1).