Macrophage-derived chemokine CCL22 and regulatory T cells in ovarian cancer patients.

The study was undertaken to evaluate macrophage-derived chemokine (CCL22) levels in the peritoneal fluid (PF) and plasma of patients with ovarian cancer (n = 93) in relation to regulatory T cells (Tregs; n = 75). The peritoneal fluid CCL22 concentrations were significantly higher in epithelial ovarian cancer (EOC) patients than in patients with benign tumors-serous cystadenoma (n = 32). There was no difference in plasma levels of CCL22 in EOC patients compared with the non-cancer and healthy volunteers (n = 10). There were no significant differences in the plasma and PF CCL22 levels based on tumor grade. However, women with stage IV FIGO (International Federation of Gynecologists and Obstetricians) had significantly higher plasma CCL22 levels than patients with stages I and III. Women with stage I FIGO had significantly higher PF CCL22 levels than patients with stages II and III. Women with endometrioid cystadenocarcinoma had higher PF CCL22 levels than women with undifferentiated carcinoma. The percentage of tumor-infiltrating Tregs (11.06 %) was significantly higher compared to PF (3.05 %) and peripheral blood (PB) (2.01 %). Moreover, the percentage of Tregs was higher in the PF than in the PB of EOC patients. There were no significant differences in the PB, PF, and tumor-infiltrating Tregs percentage based on tumor stage, grade, or histology. Elevated levels of CCL22 found in the ascites could create a chemokine gradient aiding in Treg cells migration. Increased Tregs percentage in the local microenvironment of ovarian cancer might be an important mechanism of immunosuppression.

Penetration studies of an extremely lipophilic active model substance from an oil-in-water emulsion: influence of the lipophilicity of the formulation in human skin - part 2.

The effect of the lipophilicity of a carrier on human skin penetration of an extremely lipophilic active model substance was evaluated by using Franz type diffusion cells. Oil-in-water model emulsions containing different amounts of the oily phase were prepared, and Myritol® PC (M-PC) was selected as lipophilic marker component of the oily phase. The penetrated amounts of the lipophilic model substance salicyloyl phytosphingosine (SP) were determined by high-performance liquid chromatography with ultraviolet detection, while M-PC was detected using gas chromatography coupled with mass spectrometry. It has been ascertained that the amount of the lipid phase within the emulsion influenced the penetration profile of the active ingredient SP. The emulsion containing the lowest proportion of the lipid phase provides the best conditions for SP penetration. Surprisingly, the penetration behavior of M-PC was influenced by the oily phase in the same way. Regarding the M-PC and the SP penetration profiles from each emulsion, a solvent drag mechanism can be assumed whereby M-PC acts as penetration enhancer. In conclusion, the penetration rate of the active ingredient SP and the marker component M-PC are in reverse proportion to the oil content of the formulations. The lipophilicity of SP and M-PC, their solubility and their thermodynamic activity within the vehicle could have an effect on their penetration behavior. Additionally, M-PC has the property to enhance the penetration rates of extremely lipophilic substances even at low concentrations.

Dendritic cell subsets in the peritoneal fluid and peripheral blood of women suffering from ovarian cancer.

BACKGROUND: Evaluation of immature myeloid and lymphoid dendritic cells (DCs) in the peritoneal fluid (PF) and peripheral blood (PB) mononuclears of women with ovarian carcinoma (n = 47) and benign ovarian tumors (n = 37). METHODS: Mononuclear cells were isolated from PF and PB, stained with monoclonal antibodies (mAbs) against DC antigens (anti-BDCA-1, anti-BDCA-2), and estimated using flow cytometry. RESULTS: The percentage of PF myeloid DC (MDC) in mononuclears was significantly lower in patients with ovarian cancer in comparison to the group of nonmalignant ovarian tumors (0.65% and 6.95%). The percentage of PF lymphoid DCs (LDCs) was higher in patients with ovarian cancer than in the reference group (0.64% and 0.09%). The percentage of PB MDCs and LDCs did not differ significantly between studied groups. In women suffering from ovarian cancer the percentage of both MDCs and LDCs was higher in the PF than in the PB. In the reference group the percentage of MDCs was higher but that of LDCs was lower in the PF than in the PB. In women with ovarian cancer, PF MDCs/LDCs ratio was lower in comparison to patients with serous cystadenoma. In PB the ratio of MDCs to LDCs did not differ significantly between studied groups. CONCLUSIONS: We concluded that MDCs population may be affected by the presence of malignant disease. LDC subsets may have influence on the local immune response in the PF of women with malignant tumors of the ovary. (c) 2008 Clinical Cytometry Society.

Myeloid and lymphoid dendritic cells in the peritoneal fluid of women with ovarian cancer.

PURPOSE: The aim of the study was to estimate the myeloid and lymphoid subpopulation of dendritic cells (DCs) in the peritoneal fluid (PF) of women with ovarian tumors. MATERIAL AND METHODS: We studied 34 patients with serous cystadenocarcinoma and 29 women with serous cystadenoma. Dendritic cells were isolated from peritoneal fluid, stained with monoclonal antibodies anti-BDCA-1 and anti-BDCA-2 and estimated using flow cytometry. RESULTS: Peritoneal fluid myeloid DCs constituted 0.59% of mononuclear cells in patients with ovarian cancer and 7.2% in women with serous cystadenoma. Lymphoid DCs constituted 0.39% of PF mononuclears in women with ovarian cancer and 0.07% in patients with serous cystadenoma. The percentage of lymphoid DCs was higher in patients with ovarian cancer than in women with serous cystadenoma. The BDCA-1/BDCA-2 DCs ratio in peritoneal fluid of patients with serous cystadenoma was significantly higher in comparison to ovarian cancer. CONCLUSIONS: Decreased BDCA-1/BDCA-2 DCs ratio in patients with ovarian cancer may favour Th2 lymphocyte differentiation and/or induction of immunological tolerance.

Peritoneal fluid leptin concentration in infertile patients.

Leptin may play a role in the regulation of menstrual cycle acting either directly on ovaries or at the level of the hypothalamic-pituitary axis. Peritoneal fluid is a biologically active environment that influences ovarian function but, on the other hand, concentration of many substances in peritoneal fluid can reflect the functional status of ovaries. In our study, we estimated leptin concentration in peritoneal fluid in relation to serum leptin concentration in infertile patients. A study group consisted of 31 infertile patients that underwent laparoscopy. In 15 patients, laparoscopy did not solve the problem and they were grouped as unexplained infertility, in eight patients we found endometriosis and eight patients were diagnosed as polycystic ovary syndrome (PCOS). We found significantly higher peritoneal fluid leptin concentrations in patients with unexplained infertility and endometriosis compared to those with PCOS. The plasma leptin concentration did not differ between the groups studied. There was no correlation between peritoneal fluid and plasma leptin concentrations in any patient group. Taking into account our results, the role of leptin in reproduction should be emphasized but further investigation is needed to determine its mechanism of action.

[Lipid peroxides, tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) in peritoneal fluid from infertile women with minimal and mild endometriosis]. / Produkty peroksydacji lipidów, czynnik martwicy nowotworów alpha (TNF-alpha) oraz interferon gamma (IFN-gamma) w plynie otrzewnowym nieplodnych kobiet z lagodna postacia endometriozy.

OBJECTIVES: To assess the concentrations of tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma) and lipid peroxides (the marker of free radicals activity) in peritoneal fluid (PF) of infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: 19 women were studied, including 9 infertile women with minimal or mild endometriosis and 10 patients with tubal occlusion (the reference group). Lipid peroxides (malonyldialdehyde and 4-hydroxynonenal), TNF-alpha and IFN-gamma concentrations were measured in the PF using commercially available kits. RESULTS: Concentration of IFN-gamma was detectable in PF of 7 (77.8%) women with endometriosis and in PF from 3 (30%) patients with tubal occlusion. Neither TNF-alpha or lipid peroxides PF concentration differed significantly (p < 0.05) between the groups. In the group with endometriosis we have found a positive correlation (R = 0.77, p = 0.04) between the concentrations of TNF-alpha and IFN-gamma. CONCLUSIONS: Our results suggest that oxidative stress in the PF doesn't appear to play a role in endometriosis-associated infertility.

[Concentrations of MMP-1, TIMP-1, MMP-1/TIMP-1 and I CTP complexes in follicular fluid as related to fertilization rate in women treated with in-vitro fertilization]. / Stezenie MMP-1, TIMP-1, kompleksów MMP-1/TIMP-1 i I CTP w plynie pecherzykowym w odniesieniu do wskaznika zaplodnienia komórek jajowych u kobiet leczonych metoda zaplodnienia pozaustrojowego.

OBJECTIVES: Metalloproteinases and their inhibitors appear to control connective tissue remodelling during follicular rupture. The aim of the study was to establish if oocytes fertilisation rate after ovulation induction depends on the concentrations of MMP-1, its inhibitor TIMP-1, MMP-1/TIMP-1 complex and I CTP in follicular fluid (FF). MATERIAL AND METHODS: FF were collected from 37 infertile patients undergoing ovulation induction using either short or long protocol. FF was obtained 36 hours after administration of hCG (Pregnyl). The level of MMP-1, TIMP-1, MMP-1/TIMP-1 complex were measured using ELISA kits and I CTP, E2, FSH, LH, using RIA assay kits. RESULTS: Statistically significant (p < 0.05) difference was found in TIMP-1, E2, and FSH concentration, being higher in the group with more than 75% fertilisation rate: TIMP-1 728.8 + 100.1 ng/ml vs 666.3 + 94.5 ng/ml; E2 477.3 +/- 160.0 ng/ml vs 368.0 +/- 190.0 ng/ml and FSH 7.27 +/- 1.45 mIU/ml vs 6.24 +/- 1.6 mIU/ml. CONCLUSIONS: Statistically significant increase in TIMP-1 concentration observed among patients with fertilisation rate above 75% indicates an important role of this substance in ovulation process.

Total antioxidant status of peritoneal fluid in infertile women.

OBJECTIVE: To determine whether impairment of the antioxidant systems of peritoneal fluid might be a factor responsible for infertility. STUDY DESIGN: Total antioxidant status was measured in peritoneal fluid obtained from 18 infertile women suffering from minimal or mild endometriosis, 23 patients with unexplained infertility, 12 women with tubal infertility and 13 fertile women. RESULTS: Total antioxidant status was significantly lower in peritoneal fluid from women with unexplained infertility (0.49+/-0.21 mmol/l) compared to both fertile patients (0.67+/-0.24 mmol/l, P=0.02) and women with tubal infertility (0.76+/-0.26 mmol/l, P=0.001). Peritoneal fluid total antioxidant status did not differ significantly between patients with endometriosis (0.61+/-0.2 mmol/l), tubal infertility and the fertile group (P>0.05). CONCLUSIONS: Our results suggest that low antioxidant status in peritoneal fluid may play a role in the pathogenesis of infertility.

[Peritoneal fluid and plasma 4-hydroxynonenal and malonyldialdehyde concentrations in infertile women]. / Stezenie 4-hydroksynonenalu i dialdehydu malonowego w plynie otrzewnowym i osoczu krwi nieplodnych kobiet.

OBJECTIVES: To investigate the possible role of oxidative stress in the pathogenesis of female infertility, we examined the peritoneal fluid (PF) and plasma lipid peroxides levels in infertile women. MATERIALS AND METHODS: 4-hydroxynonenal (4-HNE) and malonyldialdehyde (MDA) concentrations were measured in peritoneal fluid and plasma samples obtained from: 10 infertile women with minimal and mild endometriosis, 24 patients with unexplained infertility, 11 women with PCOS and 14 patients with tubal infertility. RESULTS: PF lipid peroxides level was significantly higher in women with idiopathic infertility compared to other groups. Plasma 4-HNE and MDA concentrations did not differ significantly between the infertile groups. No significant difference or relationship was observed between peritoneal fluid and plasma lipid peroxides levels. CONCLUSIONS: Increased free radicals activity in the peritoneal fluid environment may be the factor responsible for some cases of "unexplained infertility".

[Expression of the survivin gene in the scar endometriosis and in normal human endometrium]. / Ekspresja genu surviviny w tkance endometriozy w bliznie oraz w prawidlowym endometrium.

UNLABELLED: The survivin gene encoding a novel inhibitor of apoptosis protein (IAP) is located on chromosome (17q25). It is expressed during development and in human cancer in vivo. Endometriosis in cicatrix after cesarean section represents the only localization of endometriotic implant which etiology is known. It is believed that during the surgical procedure when the uterine cavity is open there is a high risk of decidualized endometrium implantation. The aim of the study was to estimate the expression of the survivin mRNA in the scar endometriosis after cesarean section (n = 6) and perineum endometriosis (n = 2) and in normal human endometrium (n = 12, reference group) using reverse transcribed-polymerase chain reaction (RT-PCR). METHODS: Total cellular RNA was isolated from the tissues using TriReagent according to the manufacturer's protocol. Total cellular mRNA was transcribed into cDNA with reverse transcriptase (RT) then the survivin gene fragment (430 bp) was amplified with PCR using specific primers. The products of the PCR reaction were separated on 2% agarose gel. The survivin gene was expressed in 4 of 6 cases of scar endometriosis after cesarean section and in 2/2 cases of perineum endometriosis. In all cases of normal endometrium we found the expression of survivin with a peak in the late proliferative phase. The antiapoptosis function of this gene product may play an important role in the endometriotic implant growth promotion.

[Glutathione peroxidase concentration in the peritoneal fluid from infertile women]. / Stezenie peroksydazy glutationowej w plynie otrzewnowym nieplodnych kobiet.

OBJECTIVES: To assess the concentration of Plasma Glutathione Peroxidase (plGPx) in the peritoneal fluid (PF) of patients with unexplained infertility and infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: 33 women were studied, including 8 infertile women with minimal or mild endometriosis, 15 patients with unexplained infertility and 10 patients with tubal occlusion (a reference group). Concentration of plGPx was measured in the PF using a commercially available ELISA kit (Oxis Inc.). RESULTS: The plGPx concentration was significantly (p = 0.04) lower in PF from women with unexplained infertility (846 +/- 177 ng/ml) compared to the reference group (1023 +/- 238 ng/ml), but did not differ significantly (p = 0.25) between women with endometriosis (918 +/- 81 ng/ml) and patients with tubal infertility. CONCLUSIONS: Our results suggest that low peritoneal plGPx concentration may play a role in the pathogenesis of infertility.

[Serum C-reactive protein concentration in women after hysterectomy without peritoneal suturing]. / Ocena stezenia bialka C-reaktywnego w surowicy krwi kobiet po operacjach na narzadzie plciowym bez zszywania otrzewnej.

OBJECTIVES: To assess the postoperative concentration of C-reactive protein (CRP) in serum from patients after hysterectomy with and without peritoneal suturing. MATERIALS AND METHODS: 13 women after total abdominal hysterectomy and bilateral salpingooophorectomy were studied, including 8 women with nonclosed peritoneum and 5 patients after peritoneal suturing. Concentration of CRP was measured in serum obtained before, 8, 16, 48 and 192 hours after surgery, using a commercially available ELISA kit (Eurogenetics). RESULTS: The serum concentration of CRP did not differ significantly (p < 0.05) between the groups. CONCLUSIONS: Our results suggest that nonclosure of the peritoneum layer doesn't affect the serum C-reactive protein concentration.

[Expression of the survivin gene in normal and hyperplastic endometrium]. / Ekspresja genu surviviny w prawidlowym i rozrostowym endometrium.

OBJECTIVE: Surviving a novel anti-apoptotic gene is expressed during fetal development and in numerous human cancers. MATERIAL AND METHODS: We analyzed the expression of surviving in human normal endometrium: proliferative n = 8, secretive n = 4 cases and hyperplastic endometrium 6 cases. RESULTS: RT-PCR revealed surviving expression in all cases of normal and hyperplastic endometrium. The highest expression of surviving gene was observed in hyperplastic endometrium and in the late proliferative phase.

[Total antioxidant status and activity of an extracellular superoxide dismutase in peritoneal fluid and plasma from women with unexplained infertility]. / Calkowity potencjal antyoksydacyjny oraz aktywnosc zewnatrzkomórkowej dysmutazy ponadtlenkowej w plynie otrzewnowym oraz osoczu kobiet z niewyjasniona nieplodnoscia.

OBJECTIVES: To evaluate the activity of an extracellular superoxide dismutase (EC SOD) and total antioxidant status in peritoneal fluid (PF) and plasma from women with unexplained infertility (UI). MATERIALS AND METHODS: PF and plasma samples were collected from 10 women with UI and 10 patients with tubal infertility (reference group). TAS was measured using Randox diagnostic reagent system. The activity of EC SOD was estimated using Calbiochem assay kit. RESULTS: TAS was significantly lower in PF from women with UI. Plasma TAS did not differ significantly between the groups. Peritoneal TAS was significantly lower compared to plasma TAS and there was a positive correlation between PF and plasma total antioxidant status. Activity of EC SOD did not differ significantly between the groups in either PF and plasma samples. CONCLUSIONS: These data demonstrate that low antioxidant status of PF but not that of blood may be responsible for idiopathic infertility. The activity of EC SOD doesn't appear to play a role in UI.

[Histopathological verification of the cervical pap smears based on material of the obstetrical-gynecological department of the provincial hospital in Kielce]. / Histopatologiczna weryfikacja rozmazów cytologicznych szyjki macicy.

The results of the cervical Pap. smear taken before cold knife conization and the histopathological results of the cones removed in 336 women, were compared. In cytological examination we found 10.8% (13/120) false negative results, which did not confirm of the existence the precancerous lesion within cervical epithelium. The mild dysplasia according to Pap. smears was established among 15.9% (15/94) of women with CIN III and CIN II diagnosed pathologically. In part of the women the histopathological results of the postoperative specimens and bioptic cervical material were compared. Clinical value and correlation between histopathological and cytological results in the estimation of CIN (cervical intraepithelial neoplasia) were discussed with particular regard to CIN I according to Bethesda system.

[Malonyldialdehyde and total antioxidant status in the peritoneal fluid of infertile women]. / Stezenie dialdehydu malonowego oraz calkowity potencjal antyoksydacyjny w plynie otrzewnowym nieplodnych kobiet.

OBJECTIVE: To estimate the concentration of malonyldialdehyde (MDA) and total antioxidant status in the peritoneal fluid (PF) of patients with unexplained infertility (UI) and infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: PF was obtained during laparoscopy from 8 women with UI, 12 infertile women with endometriosis (I degree and II degrees rAFS) and 10 women with benign noninflammatory ovarian tumours. All laparoscopies were performed in the follicular phase of the cycle. MDA concentration was measured according to Ledwozyw method, TAS was measured spectrophotometrically using RANDOX diagnostic reagent system. RESULTS: We found significantly higher concentration of MDA in PF from both patients with UI (p = 0.03) and with endometriosis (p = 0.046) compared to the control group. TAS was significantly (p = 0.027) higher in PF of women with UI but did not differ significantly (p = 0.49) between patients with endometriosis and controls. CONCLUSIONS: Our results show that an imbalance between lipid peroxides and the antioxidant system in PF environment may be one of the main factors responsible for the UI. In the group with endometriosis a marginally significant difference in MDA levels, no significant differences in TAS and data from the literature, suggest that accelerated lipid peroxidation in PF doesn't appear to play a role in the endometriosis associated infertility.

Regression of left ventricular hypertrophy in hypertensive patients after 1 year of treatment with rilmenidine: a double-blind, randomized, controlled (versus nifedipine) study.

OBJECTIVE: To assess the effect of 1-year treatment with rilmenidine, an oxazoline compound that exerts its antihypertensive effects through binding to imidazoline receptors in the brainstem, on left ventricular hypertrophy (LVH) secondary to essential, mild-to-moderate hypertension [supine diastolic blood pressure (DBP)95-115 mmHg]. METHODS: We performed a double-blind, randomized, controlled (versus slow-release nifedipine) trial. Adjustment of treatment took place every month (M) between inclusion (MO) and an evaluation after 6 months (M6), then during M9 and after 1 year (M12) to achieve supine DBP values < or = 90 mmHg. Patients were dropped from our study if they had DBP> 95mmHg during two consecutive visits or DBP>115 mmHg on one occasion. The daily dosage of rilmenidine was 1 mg, and could be increased to 2 mg/day. The daily dosage of slow-release nifedipine was started from the beginning at the maximum dosage of 40 mg/day, so that there was no true adjustment of treatment despite the allocation of patients to a different unit in the case of DBP> 95 mmHg. The primary criterion was the change in left ventricular mass index (LVMI, g/m2), assessed by echocardiography, between MO and M12 for patients who completed the trial. RESULTS: After a 1-month placebo run-in period, 76 patients were selected and 73 were included (35 treated with rilmenidine and 38 treated with nifedipine). Fifteen patients withdrew from the study and two completed the study with a major deviation from protocol, leaving 56 patients (24 treated with rilmenidine and 32 treated with nifedipine) for a per-protocol analysis. Baseline demographic characteristics and history of arterial hypertension for the rilmenidine and nifedipine groups were similar, for included patients and for those taken into account for the per-protocol analysis. Between MO and M12, DBP in members of the per-protocol population was adequately controlled for those in the rilmenidine group (102.7+/-4.6 versus 88.5+/-7.1 mmHg, respectively) and for those in the nifedipine group (102.7+/-5.1 versus 85.6+/-79 mmHg, respectively). During MO, LVMI of patients in the rilmenidine group (176.9+/-41.3 g/m2) was slightly higher than that of patients in the nifedipine group (172.6+/-35.1 g/m2). During M12, LVMI was observed to have decreased both for patients in the rilmenidine group (to 154.8+/-40.2 g/m2, a decrease of 22.1+/-23.3 g/m2, P< 0.001) and for those in the nifedipine group (to 145.6+/-36.4 g/m2, a decrease of 26.9+/-29.5 g/m2, P< 0.001) but the difference between these two groups was not significant (P= 0.5). CONCLUSION: One-year treatment with a daily dosage of 1 or 2 mg rilmenidine achieves a significant reduction of left ventricular mass, which is not statistically different than that occurring with a daily dosage of 40 mg of slow-release nifedipine.

[Multiple myeloma nephropathy as a cause of renal failure--diagnostic difficulties and prospects for treatment]. / Nefropatia szpiczakowa jako przyczyna niewydolnosci nerek--trudnosci diagnostyczne i mozliwosci leczenia.

We describe two patients admitted to our hospital because of renal failure. We diagnosed multiple myeloma in both. One patient had high peripheral blood eosinophilia, that normalized during therapy. The patients were treated with melphalan and prednisone, hemodialysis and one patient with recombinant human erythropoietin. Both patients responded to that treatment: after 7 months the number of plasma cell in bone marrow decreased from 65 to 10% in the first patient, and from 38 to 4% in the second patient. They returned to work and were on maintenance hemodialysis 2 times weekly. The patients have been observed for 21 months.