The Potential of Asiatic Acid in the Reversion of Cyclophosphamide-Induced Hemorrhagic Cystitis in Rats.

The purpose of this study was to determine if asiatic acid may act efficiently in the model of cyclophosphamide (CYP)-induced cystitis in rats. We performed experiments after administration of CYP (single dose 200 mg/kg, intraperitoneally), asiatic acid (30 mg/kg/day for 14 consecutive days, by oral gavage), or CYP plus asiatic acid, during which conscious cystometry, measurements of urothelium thickness and bladder edema, as well as selected biomarkers analyses were conducted. In rats that received asiatic acid together with CYP, a drop in bladder basal pressure, detrusor overactivity index, non-voiding contraction amplitude, non-voiding contraction frequency, and the area under the pressure curve were observed, when compared to the CYP group. Furthermore, a significant increase in threshold pressure, voided volume, intercontraction interval, bladder compliance, and volume threshold to elicit NVC were found in that group accordingly. Administration of the asiatic acid successfully restored concentrations of biomarkers both in bladder urothelium (BDNF, CGRP, OCT-3, IL-1ß, IL-6, NGF, nitrotyrosine, malondialdehyde, TNF-α, SV2A, SNAP23, SNAP25, PAC-1, ORM1, occludin, IGFBP-3, HB-EGF, T-H protein, Z01, and HPX) and detrusor muscle (Rho kinase and VAChT) in CYP-treated rats. Finally, asiatic acid significantly decreased urothelium thickness and bladder oedema. Asiatic acid proved to be a potent and effective drug in the rat model of CYP-induced cystitis.

Peroxiredoxin-1 as a prognostic factor in patients with ovarian cancer.

INTRODUCTION AND OBJECTIVE: Peroxiredoxin-1 (PRDX-1) belongs to a family of antioxidant enzymes and has proved to be a versatile molecule regulating cell growth, differentiation and apoptosis. PRDX1-regulated signaling pathways play an important role in the progression and metastasis of human tumours, especially in breast, esophageal and lung cancers. The aim of the study was to evaluate the expression of PRDX-1 in ovarian cancer tissues, and to test the clinical value of PRDX-1 as a prognostic factor in this malignancy. MATERIAL AND METHODS: PRDX-1 expression was assessed by automated immunohistochemistry in tumours taken from 55 patients with ovarian cancer during primary surgery. Specimen were formalin-fixed and preserved in paraffin-embedded blocks. The results were correlated with clinicopathological data. RESULTS: A high expression of PRDX-1 was observed in 20% of cases, and was associated with worse compliance to chemotherapy protocol (P<0.002), worse response to chemotherapy (P<0.04), and higher levels of CA 125 after the 1st line treatment (P<0.004). PRDX-1 positive subjects had a significantly lower 5-year disease-free survival (9.1% vs. 42.6%, P<0.01) and a lower 5-year overall survival (9.1% vs. 56.7%; P<0.002). Multivariate analysis showed that a high expression of PRDX-1 is an independent prognostic factor of poor, overall survival (P<0.002) and a disease-free survival (P<0.01). CONCLUSIONS: Results of the study show that PRDX-1 expression in tumour tissues can be another biomarker of prognosis in patients with ovarian cancer.

Prognostic value of tissue plasminogen activator (tPA) in patients with epithelial ovarian cancer undergoing chemotherapy.

OBJECTIVES: Tissue plasminogen activator (tPA) is a key enzyme for fibrin degradation and the proteolytic defense against formation of the thrombotic endothelial deposits. tPA is involved in carcinogenesis but its exact role in tumor biology is not very well understood and a prognostic value of tPA remains ambiguous in different cancers. The aim of the study was to assess the prognostic value of plasma tPA in patients with epithelial ovarian cancer (EOC) in the course of the first line chemotherapy. MATERIAL AND METHODS: the study covered 60 patients with EOC who underwent the 1st line chemotherapy. Plasma tPA was assessed at onset, after 3 and 6 cycles of chemotherapy. The groups were stratified according to tPA level at onset of chemotherapy (low tPA group < 6.5 mg/L, N = 37 and high tPA group > 6.5 mg/L, N = 23). Survival analysis was repeated for the cut-off of tPA level at 6.5 mg/L and 5.1 mg/L after 3 and 6 cycles. RESULTS: Only subjects with tPA > 6.5 mg/L at onset of chemotherapy had a significantly lower probability of a 5-year survival (34.8% vs. 72.7%, P < 0.006) and lower chance for disease free survival within 5 years (39.3% vs. 72.7%, P < 0.014). tPA < 6.5 mg/L plasma level evaluated at onset of chemotherapy was an independent marker of better overall survival (RR = 0.44, 95%CI = 0.19-0.98) but not disease-free survival. CONCLUSIONS: Plasma tPA may serve as a marker of survival if assessed at onset of the first line chemotherapy in patients with ovarian cancer.

Plasminogen Activator Inhibitor Type 1 in Blood at Onset of Chemotherapy Unfavorably Affects Survival in Primary Ovarian Cancer.

Plasminogen activator inhibitor type 1 (PAI-1) belongs to the family of the plasminogen activator system. PAI-1 stimulates fibrinolysis and also promotes tumor progression. The aim of this study was to evaluate the prognostic value of blood plasma PAI-1 content in patients with epithelial ovarian cancer who start the first-line chemotherapy. PAI-1 content was measured in the blood of 61 patients with epithelial ovarian cancer at onset of first-line chemotherapy. The patients were further stratified into the low PAI-1 group (≤20 ng/mL; 33 patients) and the high PAI-1 group (>20 ng/mL; 28 patients). We found that the greater plasma PAI-1 content was associated with a significantly lower probability of a 5-year-long survival compared to that when PAI-I content was lower (45.5% vs. 69.5%, respectively; p = 0.04). However, the risk of cancer recurrence within 5 years failed to differ appreciably. A multivariate analysis revealed that the lower PAI-1 plasma content was an independent factor of longer overall survival (death risk ratio of 0.36, 95%CI = 0.16-0.79; p < 0.01). We conclude that PAI-1 is yet another biomarker of survival in patients with ovarian cancer.

Peroxiredoxin-5 is a negative survival predictor in ovarian cancer.

OBJECTIVES: Peroxiredoxins (PRDXs) constitute a family of antioxidant enzymes which are also involved in the process of carcinogenesis. They are composed of six identified isoforms (PRDX-1-6) and are supposed to play different roles in tumor progression, depending on type of cancer and member of the PRDX family. The aim of the study was to assess the prog- nostic value of PRDXs in ovarian cancer. MATERIAL AND METHODS: a dataset of patients with ovarian cancer from The Cancer Genome Atlas was analyzed. Expression of PRDX-1 to 6 mRNA was evaluated in 260 samples. The prognostic value of PRDXs was assessed using the Cox regression model which included the following clinical and pathological data: age, clinical stage, tumor grade, and residual disease. RESULTS: Within the PRDXs family, only higher expression of PRDX-5 was associated with worse overall survival both, in unselected patients and > 50-year-olds. PRDX-5 expression and residual disease were independent negative prognostic factors of patient survival. CONCLUSIONS: PRDX-5 is a negative predictor of survival in ovarian cancer.

Magnetic resonance diffusion-weighted imaging in diagnostics of primary fallopian tube carcinoma - is it useful?

PURPOSE: Primary fallopian tube carcinoma (PFTC) is the rarest form of female genital malignancy. The imaging applied for suspected adnexal masses includes transvaginal ultrasound (US), computed tomography (CT), and magnetic resonance imaging (MRI), but the vast majority of PFTC is recognised intraoperatively. MATERIAL AND METHODS: The study group consisted of seven women with postoperatively histopathological diagnosis of PFTC. To recognise characteristic findings for PFTC, retrospective analysis of preoperative MRI was performed. All patients underwent MRI of the pelvis and abdomen using a 1.5T MR system. Based on the results of the above imaging, suspected adnexal masses were recognised. MRI protocol contained T2-weighted images, fat-suppressed T2-weighted, T2-TIRM, DW EPI, pre- and postcontrast dynamic 3D T1 GRE in transverse orientation, with diffusion weightings of 0, 50, 100, 150, 200, 400, 800, and 1200 s/mm2. Regions of interest were outlined by a radiologist, who documented the character of adnexal masses on diffusion-weighted (DW) images and apparent diffusion coefficient (ADC) maps. RESULTS: In all seven patients with PFTC unilateral tumour was found. On all DW images (with ß values of 0, 50, 100, 150, 200, 400, 800, and 1200 s/mm2) the mean signal intensities of solid parts of tumour were significantly higher than the mean signal intensities of normal ovarian tissue (p = 0.0001). There were no statistically significant differences between eight ß values applied for ADC calculations. CONCLUSIONS: Preoperative diagnostics of PFTC is difficult and mainly based on morphological features. Previous research did not show characteristics of PFTC in post-contrast dynamic imaging. In our material a clear increasing of signal intensity in DW imaging occurred independently of the ß value.

Potentialization of N-a-tosyl-L-phenylalanine chloromethyl ketone (TPCK) cytotoxic activity by 2-(1-adamantylamino)-6-methylpyridine (AdAMP) in human ovarian cancer cells.

OBJECTIVES: TNF is one of the key cytokines involved in cancer development. TNF signaling can result in both stimulating and inhibitory signals that can result in opposite biological effects in cancerogenesis. 2-(1-adamantylamino)-6-methylpyridine (AdAMP) enhances TNF secretion whereas N-a-tosyl-L-phenylalanine chloromethyl ketone (TPCK) is a NF-κB inhibitor potentially stimulating proapoptotic TNF signals. The aim of the study was to assess the effect of TPCK in combination with AdAMP on human ovarian cells. MATERIAL AND METHODS: CAOV-1 human ovarian cell line was incubated with TPCK and AdAMP for 24 hours. The cytotoxic effect was evaluated in a crystal violet assay. A monoclonal antibody against TNF, Infliximab, was added to examine the possible mechanism of interactions. RESULTS: Depending on concentration, AdAMP potentialized cytotoxic activity of TPCK or had a synergistic effect with TPCK. Infliximab did not reverse cytotoxicity of AdAMP and TPCK and in some cytotoxic and non-cytotoxic concentrations even enhanced their cytotoxicity. CONCLUSIONS: AdAMP and TPCK cytotoxicity seems to be dependent on TNF signaling, however, the exact mechanism of interactions remains unclear.

Results of optimal debulking surgery with bowel resection in patients with advanced ovarian cancer.

BACKGROUND: The surgical treatment of patients with advanced-stage ovarian cancer is based on maximal cytoreduction with widening the debulking on the extra-ovarian tissues and infiltrated organs. The purpose of the study was to assess the outcome after optimal cytoreduction with partial bowel resection and to find the risk factors of relapse. Another goal was the quantitative and qualitative assessment of intra- and postoperative complications in the studied group. METHODS: The analysis of debulking procedures with intestinal resection and postoperative period in 33 ovarian cancer patients, The International Federation of Gynecology and Obstetrics (FIGO) stages III and IV, was performed. RESULTS: The optimal cytoreduction defined as less than 1.0 cm residual disease was achieved in all patients including the following: 26 patients (78.8%) with no macroscopic residual disease, 4 patients (12.1%) with the largest residual tumor less than 0.5, and 3 patients (9.1%) with 0.5 cm to less than 1.0 cm residual disease. The rectosigmoid resection was the most common surgical procedure (n = 27). The risk of relapse was significantly higher in subjects who had the macroscopic residual tumor left during the primary operation (57.1 vs. 11.5%, P = 0.035). A primary bowel tumor size was another predictor of relapse. The maximum tumor diameter was significantly larger (14.9 ± 6.7 cm vs. 10.3 ± 4.7 cm, P = 0.047) in patients who developed the relapse. CONCLUSIONS: As presented in the article, our outcomes and other authors' observations indicate that debulking surgery with bowel resection in patients with advanced ovarian cancer brings good results. Complications connected with bowel surgery are to be accepted. The interesting thing is that a primary bowel tumor size was a predictor of relapse.

[Splenectomy as a part of debulking surgery in patients with advanced ovarian cancer]. / Splenektomia jako czesc operacji cytoredukcyjnej u pacjentek z zaawansowanym rakiem jajnika.

OBJECTIVES: The aim of the study was the assessment of perioperative complications in patients with advanced ovarian cancer who underwent splenectomy to achieve optimal debulking. MATERIAL AND METHODS: We analyzed eight debulking procedures with splenectomy and the postoperative period in ovarian cancer patients, FIGO stage III/B-IV. Preoperative diagnostics included multidetector computed tomography (MDCT) or diffusion-weighted echo-planar magnetic resonance (MR-DWI). The following factors were analyzed: size of the removed tumor, size of remains left, blood loss, packed red blood cell transfusion, quantity and reason for reoperations, pancreatic amylase concentrations in the drainage fluid, wound infection, fever over 38 degrees C, and length of hospitalization. RESULTS: Complete debulking was achieved in 8 patients, including 5 cases with no macroscopic residual lesions and 3 patients with lesion diameter of < 10 mm. Median operative time was 175 min. There was one case of reoperation caused by perforation of the stomach wall (histologically confirmed stress ulcer). Median blood loss was 1050 ml and the rate of packed red blood cells transfusion was 75%. Elevated amylase levels in the drainage fluid was noted in 6 patients. Amylase concentration was greater than 5 times the normal serum value during the first postoperative day. After postoperative day 3 it was lower than normal serum range. There were no cases of postoperative fever wound infections, or deaths. The length of hospitalization was 6 days. CONCLUSIONS: Splenectomy as a part of cytoreductive surgery for advanced ovarian cancer may contribute to achieving complete debulking and bring benefits, especially in cases with no macroscopic residual disease. The risk of intra- and postoperative complications related to splenectomy seems to be acceptable.

Synergistic cytotoxic effect of sulindac and pyrrolidine dithiocarbamate against ovarian cancer cells.

Sulindac, a non-steroidal anti-inflammatory drug, suppresses carcinogenesis and inhibits growth of tumor cells. Pyrrolidine dithiocarbamate (PDTC), a potent NF-κB inhibitor, has been also identified as a potential anti-neoplastic agent. We hypothesized that combination of sulindac and PDTC could result in augmentation of cytotoxicity against ovarian cancer cells. The effect of sulindac and PDTC was examined on several ovarian cancer lines. Tumor cell viability was assessed using the MTT assay. Annexin-V/PI staining was used to detect apoptosis, cell cycle distribution was analyzed in FACS, and expression of cellular proteins was detected by western blotting. Incubation of OVA-14, OVP-10 and CAOV-1 ovarian cancer cells with sulindac and PDTC resulted in significantly greater inhibition of cell viability compared to either compound alone. In a model of OVA-14 cells it was evident that this effect was not related to the expression of COX enzymes since both active (sulindac sulfide) and inactive (sulindac) in vitro compounds affected the growth of tumor cells to a similar extent and synergized in cytotoxicity with PDTC. Combination of sulindac and PDTC lead to G0 arrest and massive apoptosis in co-treated cultures. Western blotting analysis argued for induction of the mitochondrial apoptotic pathway. These data demonstrate the synergistic cytotoxic effect of sulindac and PDTC on ovarian cancer cells through apoptosis and cell cycle arrest and prompt to test the efficacy of this combination in animal models.

[Antitumor effects of sulindac in ovarian cell cultures]. / Przeciwnowotworowy efekt sulindaku w hodowlach komórek raka jajnika.

OBJECTIVE: The purpose of our study was to assess susceptibility of cells of various ovarian cell lines on different nonsteroidal anti-inflammatory drugs (NSAIDs). MATERIALS AND METHODS: Cytotoxic effect of NSAIDs was tested using MTT colorimetric assay. RESULTS: Amongst 6 NSAIDs tested: sulindac, sulindac sulfide, sulindac sulfone, acetylsalicylic acid, nimesulide, and rofecoxib, viability of ovarian carcinoma cells was compromised most strongly by sulindac and sulindac sulfide and concerned all the cell lines tested: SKOV-3, MDAH 2774, OVCA-1, and OVP-10. Sulindac sulfone and rofecoxib also displayed some cytotoxic effect during prolonged 72-hour incubation. Other NSAIDs tested: nimesulide and acetylsalicylic acid were devoid of cytotoxic effect on ovarian cancer cells. CONCLUSION: Our results are encourage enough to conduct clinical trials that could allow to draw conclusions regarding potential application of sulindac in the adjuvant treatment of a standard chemotherapy of ovarian cancer.

Uteroplacental circulation in early pregnancy complicated by threatened abortion supplemented with vaginal micronized progesterone or oral dydrogesterone.

OBJECTIVE: To compare the influence of vaginal micronized progesterone and oral dydrogesterone supplementation on uteroplacental circulation in early pregnancy that is complicated by threatened abortion. DESIGN: Randomized, parallel group, double-blind, double dummy-controlled study. SETTING: Tertiary care university hospital. PATIENT(S): Fifty-three patients with threatened abortion and a living embryo. INTERVENTION(S): Three hundred milligrams of micronized vaginal progesterone or 30 mg of oral dydrogesterone daily supplementation for 6 weeks, serial transvaginal Doppler ultrasound measurement of pulsatility index, resistance index, and systolic/diastolic ratio of the spiral arteries, the uterine arteries, and the intrachorionic area. MAIN OUTCOME MEASURE(S): Uteroplacental blood flow. RESULT(S): The study demonstrated that vaginal progesterone administration, but not oral dydrogesterone treatment, results in the decrease in the spiral artery pulsatility and resistance index and systolic/diastolic ratio. Insignificant decrease in pulsatility index and resistance index of the uterine artery was observed at >9 weeks and was not associated with treatment regimen. Dydrogesterone treatment was only accompanied by the decrease in the uterine artery systolic/diastolic ratio. CONCLUSION(S): Vaginal progesterone and oral dydrogesterone supplementation have a different influence on the uteroplacental circulation in early pregnancy that is complicated by threatened abortion.

TNF-alpha production-enhancing activity of 2-(1-adamantylamino)-6-methylpyridine (AdAMP) in cultures of human normal and neoplastic cells.

The purpose of this study was to determine the TNF-alpha-stimulatory effect of a novel immunomodulator 2-(1-adamantylamino)-6-methylpyridine (AdAMP) on normal and neoplastic human cells. In a panel of several human ovarian cancer cell lines, almost half of them spontaneously secreted significant amounts of TNF-alpha. When incubated with AdAMP, a 3-fold enhancement of TNF-alpha production by cells was observed. Furthermore, the phorbol myristic acetate ester (PMA)-induced release of TNF-alpha in cultures of U937 cells was increased in the presence of AdAMP. Primary monocytes isolated from peripheral blood did not respond to AdAMP. Although cytokine release was not triggered in human peripheral blood monocytes, AdAMP co-stimulated these cells to produce TNF-alpha and IL-8 during incubation with lipopolysaccharide (LPS). No effect of AdAMP was found on IL-1beta and IL-6 production by monocytes. In cultures of peripheral blood T lymphocytes, AdAMP significantly decreased the adhesion of these cells to matrix proteins in an in vitro assay. The results suggest that AdAMP, as a stimulator of cytokine secretion, may have potential application in tumor therapy.

Effective photoimmunotherapy of murine colon carcinoma induced by the combination of photodynamic therapy and dendritic cells.

PURPOSE: The unique mechanism of tumor destruction by photodynamic therapy (PDT), resulting from apoptotic and necrotic killing of tumor cells accompanied by local inflammatory reaction and induction of heat shock proteins (HSPs), prompted us to investigate the antitumor effectiveness of the combination of PDT with administration of immature dendritic cells (DCs). EXPERIMENTAL DESIGN: Confocal microscopy and Western blotting were used to investigate the influence of PDT on the induction of apoptosis and expression of HSP expression in C-26 cells. Confocal microscopy and flow cytometry studies were used to examine phagocytosis of PDT-treated C-26 cells by DCs. Secretion of interleukin (IL)-12 was measured with ELISA. Cytotoxic activity of lymph node cells was evaluated in a standard (51)Cr-release assay. The antitumor effectiveness of PDT in combination with administration of DCs was investigated in in vivo model. RESULTS: PDT treatment resulted in the induction of apoptotic and necrotic cell death and expression of HSP27, HSP60, HSP72/73, HSP90, HO-1, and GRP78 in C-26 cells. Immature DCs cocultured with PDT-treated C-26 cells efficiently engulfed killed tumor cells, acquired functional features of maturation, and produced substantial amounts of IL-12. Inoculation of immature DCs into the PDT-treated tumors resulted in effective homing to regional and peripheral lymph nodes and stimulation of cytotoxic activity of T and natural killer cells. The combination treatment with PDT and administration of DCs produced effective antitumor response. CONCLUSIONS: The feasibility and antitumor effectiveness demonstrated in these studies suggest that treatment protocols involving the administration of immature DCs in combination with PDT may have clinical potential.

Stimulation of TNF-alpha production by 2-(1-adamantylamino)-6-methylpyridine (AdAMP) - a novel immunomodulator with potential application in tumour immunotherapy.

The immunomodulatory effects of a recently synthesized adamantane derivative of aminopyridine - 2-(1-adamantylamino)-6-methylpyridine (AdAMP) - were tested on normal and neoplastic cells in vitro. When incubated with TNF-alpha gene-transduced mouse melanoma cells (B78/TNF), AdAMP significantly enhanced basal production of TNF-alpha by these cells, both by "high" and "moderate" TNF-alpha-producer cells. A similar TNF-alpha production-enhancing effect was observed in cultures of human ovarian carcinoma cells (CAOV1) which spontaneously produce TNF-alpha but not in cultures of tumour cells incapable of TNF-alpha secretion. RT-PCR analysis showed that the enhancement of TNF-alpha production by AdAMP was associated with an increase in TNF-alpha mRNA expression in the treated cells. The results of an electrophoretic mobility shift assay (EMSA) showed that AdAMP significantly activated nuclear factor kappaB (NF-kappaB) in both CAOV1 and B78/TNF cells. The role of NF-kappaB in enhancement of TNF-alpha production was confirmed in experiments in which MG132, an inhibitor of NF-kappaB activation, reversed the effect of AdAMP. Unexpectedly, dexamethasone, a potent antiinflammatory agent and a strong inhibitor of TNF-alpha production in vivo, increased both spontaneous and AdAMP-augmented production of TNF-alpha in in vitro cultures of ovarian carcinoma cells and B78/TNF cells. AdAMP also enhanced TNF-alpha secretion by LPS-induced monocytes. AdAMP-induced augmentation of TNF-alpha production by B78/TNF cells was accompanied by morphological changes in the treated cells and a decrease in their adherence to fibrinogen and collagen IV. In view of these properties, AdAMP seems to be a therapeutically promising compound with potential application as an adjuvant augmenting the efficacy of cancer vaccine-based therapies or in the local treatment of certain tumours.