Diagnostic accuracy of interleukin-6 levels in peritoneal fluid for detection of endometriosis.

PURPOSE: To determine, with extended receiver operating characteristic (ROC) curve analysis, the diagnostic value of cytokines showing significantly different peritoneal concentrations between women with and without endometriosis. METHODS: Multiplex cytokine concentration measurement of IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ levels in peritoneal fluid of women with minimal to mild (n = 10) and moderate to severe (n = 26) endometriosis, and 42 controls. RESULTS: Only IL-6 and IL-10 concentrations were significantly higher in endometriosis patients than in controls. Specifically, significantly higher IL-6 and IL-10 levels were found in moderate to severe but not in minimal to mild endometriosis as compared to controls. For evaluation of diagnostic significance, ROC analysis determined discriminating parameters for IL-6, while those calculated for IL-10 were useless. Importantly, ROC analysis for IL-6 levels limited to women with moderate to severe endometriosis showed the highest area under the curve with the sample size sufficient to achieve 90 % power of the test. Finally, extended ROC including cost of analysis for this group of patients determined the optimal cut-off leading to high specificity and positive likelihood ratio resulting in 79 % effectiveness of the test. CONCLUSIONS: While our outcomes show moderate usefulness of peritoneal IL-6 levels in discrimination of moderate to severe endometriosis, further studies might be needed to determine the usefulness of peritoneal IL-6 levels in detection of early stages of endometriosis, as such a finding would be more relevant in clinical decision making.

Occurrence of Mycoplasma genitalium in fertile and infertile women.

OBJECTIVE: To determine the frequency of occurrence of Mycoplasma genitalium in the reproductive organs of infertile women in comparison with a control group of healthy, fertile women. DESIGN: Prospective study. SETTING: Gynecology Clinic at the 2nd Department of Gynecology and Obstetrics of the Wroclaw Medical University, Poland. PATIENT(S): The study included 51 patients with primary infertility (24 women with idiopathic infertility) and 23 women with proven fertility. INTERVENTION(S): Cervical smear and smear from the peritoneal cavity, performed during laparoscopy. MAIN OUTCOME MEASURE(S): Presence of the genetic material of M. genitalium in the collected material analyzed using polymerase chain reaction (PCR). RESULT(S): M. genitalium was found in the cervical canal of 19.6% of all infertile patients and in 4.4% of fertile patients. In addition, the pathogen was discovered in the cervical canal of 29% patients with unexplained (idiopathic) infertility, which in comparison with the fertile group was a statistically significant difference. In the abdominal cavity, M. genitalium was found in 5.8% of patients from the infertile group (in 8.4% patients with idiopathic infertility), whereas it was not detected in the material obtained from the studied fertile patients. CONCLUSION(S): The results obtained may suggest that M. genitalium is a species having an impact on impaired fertility in women.

Serum and peritoneal evaluation of vitamin D-binding protein in women with endometriosis.

INTRODUCTION: Vitamin D-binding protein (also called DBP or Gc-globulin) is recognized as a multifunctional protein involved in the action scavenger system, the transport of vitamin D sterols, and the modulation of immune and inflammatory responses. This study evaluated total serum and peritoneal concentrations of vitamin D-binding protein in women with endometriosis, known as an inflammation-associated disease. MATERIALS/METHODS: The total concentration of DBP was measured with an enzyme-linked immunosorbent assay (ELISA) using a polyclonal antibody raised in a goat immunized with human DBP. Serum and peritoneal fluid were collected from women with endometriosis (n=26) and from patients with benign gynecological conditions serving as a control group (n=17). RESULTS: In general, the vitamin D-binding protein concentration was higher in serum than in peritoneal fluid. Women with endometriosis had higher serum but lower peritoneal levels of DBP compared with the control group; however, no significance was noted. When the endometriosis group was divided with regard to severity, an insignificantly higher serum level of DBP was observed in advanced endometriosis compared with the mild form of the disease, whereas the peritoneal concentration was not dependent on disease severity. CONCLUSIONS: It is concluded that serum and peritoneal DBP concentrations are not affected in women with endometriosis; however, based on the latest published data, it is possible that both the serum and peritoneal concentrations of vitamin D-binding protein may be dependent on Gc genotype, which results in differential modulation of monocyte/macrophage activity.

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.

[Frequency of detection of Ureaplasma urealyticum and Mycoplasma hominis in cervical canal and the Douglas pouch of infertile and fertile women]. / Czestosc wykrywania Ureaplasma urealyticum i Mycoplasma hominis w kanale szyjki macicy i jamie otrzewnej u plodnych i nieplodnych kobiet.

The group of organisms commonly referred to as genital mycoplasmas comprise species most often found in genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can possibly cause many different pathologies like: non-gonococcal urethritis, bacterial vaginosis, cervicitis, endometritis or pelvic inflammatory disease. The problem of their morbidity and the possible influence they have on human fertility is still not clear. The aim of this study was to find out whether two investigated species- Ureaplasma urealyticum and Mycoplasma hominis can be detect more often in a group of infertile women. 74 women participated in the study and were assigned to one of 2 groups of patients: infertile women and fertile women without any sign of genital tract infection. Swabs from the cervical canal of the uterus and the fluid from the Douglas pouch were taken during the gynecological examination and laparoscopic procedure. Two diagnostic methods were used: biochemical method- commercial diagnostic kit- Mycoplasma IST 2 and PCR method. The results showed that Ureaplasma urealyticum and Mycoplasma hominis were detected among both fertile and infertile women with nearly the same frequency, much more often in cervical canal than in the Douglas pouch. Ureaplasma urealyticum was more common pathogen than Mycoplasma hominis in both groups and locations. The achieved results point out that the role of genital mycoplasmas in human infertility is still unclear and require further investigations.

Bicornuate rudimentary uterine horns with functioning endometrium and complete cervical-vaginal agenesis coexisting with ovarian endometriosis: a case report.

OBJECTIVE: To report a patient with bicornuate rudimentary uterine horns with functioning endometrium and complete cervical-vaginal agenesis coexisting with ovarian endometriosis. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 37-year-old woman with an extremely rare müllerian anomaly of the uterus and vagina coexisting with ovarian endometriosis. INTERVENTION(S): Resection of the rudimentary uterine horns with bilateral salpingo-oophorectomy. MAIN OUTCOME MEASURE(S): Relief from pelvic pain after the operative procedure. RESULT(S): The definite diagnosis and removal of the müllerian anomaly and endometriosis foci. CONCLUSION(S): Cyclic pelvic pain since the age of 14 was due to cryptomenorrhoea in the presence of the bicornuate rudimentary uterine horns with functioning endometrium and cervical-vaginal agenesis. Ovarian endometriosis developed as a result. In such cases, invasive procedures, such as laparoscopy or laparotomy, should be considered to establish the diagnosis. Removing the functioning rudimentary uterine horns just after menarche should prevent the development of endometriosis and hematometra.

[The presence of Mycoplasma hominis and Ureaplasma urealyticum in the cervical canal of uterus]. / Obecnosc Mycoplasma hominis i Ureaplasma urealyticum w kanale szyjki macicy kobiet.

OBJECTIVES: Mycoplasma hominis and Ureaplasma species are common commensal inhabitants of the lower genitourinary tract in adolescents and adults who are sexually active. A lot of authors points out that these microorganisms can play an important role in pathology of genital tract like pelvic inflammatory disease, sterility or non-gonococcal urethritis. MATERIALS AND METHODS: In the study samples from cervical canal of the uterus were obtained from 222 women. The first group consist of 132 women who were examined in II Gynecological Clinic in Wroclaw for different, probably infectious, gynecological pathologies (adnexitis, sterility, bacterial vaginosis). 90 women without infectious diseases were in a control group. All swabs taken from cervix were tested for Mycoplasma hominis and Ureaplasma urealyticum. RESULTS: The prevalence of Ureaplasma urealyticum was 31.8% in the first tested group (42 of 132 women were positive) and 8.8% in control group (8 of 90 were positive). 3% (4 of 132) of patients were positive to Mycoplasma hominis in the first group and only 1.1% (1 of 90) in a control group. CONCLUSIONS: Ureaplasma urealyticum was found most often in such genital tract pathologies like acute or recurrent adnexitis, sterility or bacterial vaginosis. No statistically significant correlation was found between the age of the patients and the incidence of mycoplasmas.

The coexistence of ERBB2, INT2, and CMYC oncogene amplifications and PTEN gene mutations in endometrial carcinoma.

PURPOSE: To evaluate the frequency of ERBB2, INT2 and CMYC oncogene amplifications and their coexistence with PTEN gene mutations in endometrial carcinomas. METHODS: In 54 endometrial carcinomas amplification of ERBB2, INT2 and CMYC was determined using differential polymerase chain reaction (dPCR), and mutations in all exons of PTEN were investigated by PCR-SSCP and direct sequencing methods. Results were correlated with clinicopathological features of tumors. RESULTS: In 31 out of 54 endometrial carcinomas (57.4%) genetic defects were found within the examined genes. Of all identified defects, mutations in PTEN and the amplification of CMYC were most frequent (26/54-48.1% and 10/54-18.5%, respectively). INT2 was amplified in 5.6% (3/54) of cases. In no case did we find ERBB2 amplification. In 77.4% (24/31) of cases only one gene was damaged. Of these, 20 cases showed only PTEN mutations, three cases only CMYC, and one case only INT2 amplification. In another seven out of 31 tumors (22.5%) defects in two or three genes coexisted simultaneously: PTEN and CMYC in five cases, CMYC and INT2 in one case, and PTEN, CMYC, and INT2 in one case. We found a number of interesting relations between the location of mutations within the PTEN sequence and the presence (+) or lack (-) of CMYC amplification. In the PTEN+CMYC- tumors the PTEN mutations were most frequent in exons 1-5, and less frequent in exons 7-8 (66.7% and 33.3%, respectively). In contrast, in the PTEN+CMYC+ carcinomas the PTEN mutations were found mainly in exons 7-8 (85.7%). PTEN mutations were equally frequent in both early and more advanced endometrial carcinomas. The CMYC amplification, however, was more frequent in advanced as compared to early tumors, although this difference did not reach statistical significance. CONCLUSIONS: Our data suggest that differences in the presence of genetic defects may reflect the different molecular pathways of endometrial carcinogenesis. These data also suggest that location of intragenic PTEN mutations and their coexistence with the CMYC amplification may play a crucial part in the development of various subtypes of endometrial carcinoma, but this preliminary suggestion requires further research.

[Ovarian cancer in a pregnant woman and limited treatment strategy--a case report]. / Rak jajnika u ciezarnej a oszczedzajace postepowanie terapeutyczne--opis przypadku.

We present the case of 31-year-old woman in 13th week of pregnancy with the diagnosis of the ovarian cancer (cystadenocarcinoma papillare serosum G1) in the FIGO stage Ia. Taking into consideration this data and the patient's wish of preserving the pregnancy and the childhood bearing possibility in the future we performed only left salpingooophorectomy. Postoperative and further pregnancy course was uncomplicated. The pregnancy was terminated in the 40th week by cesarean section--a well doing male newborn in term was delivered. The cesarean section was at the same time the second look procedure. All organs of peritoneal cavity were inspected--no macroscopic changes were observed, oncologic smears and specific biopsies were taken showing no neoplasmatic lesions present in the histology. Nowadays it is the 5th year of systematic patient's control, who is in complete remission. This case report is the example of the limited treatment strategy success based on prognosing factors (tumor histology, grading, staging) as well as patient's wishes of preserving the fertility.

[Course of pregnancy in a patient with systemic lupus erythematosus--case report]. / Przebieg ciazy u pacjentki z toczniem ukladowym--opis przypadku.

Authors describe a successful pregnancy outcome after prednisone and aspirin therapy in 19-year-old woman with systemic lupus erythematosus with the history of recurrent urinary tract infections, pericarditis and encephalitis.

[Evaluation of expression and correlation between P53, BCL-2 and BAX proteins in ovarian neoplasms]. / Ocena ekspresji i wzajemnych zaleznosci pomiedzy bialkami P53, BCL-2 i BAX w rakach jajnika.

OBJECTIVES: Interactions between BCL-2 and other proteins regulating programmed cells death in ovarian carcinomas are poorly understood. DESIGN: The evaluation of expression of P53, BCL-2 and BAX proteins in ovarian carcinomas. The associations between oncoproteins studied and the histological structure, grade of differentiation and stage of disease were also analysed. MATERIAL AND METHODS: The expression of P53, BCL-2 and BAX was evaluated by immunoperoxidase technique (PAP) in tissue sections and corresponding cyst and/or ascitic fluid cells in individual patients with ovarian carcinoma. RESULTS: It was shown that the expression of P53 and BAX was comparable in tissue sections and cyst or ascitic fluid cells of individual patients, however the presence of BCL-2 was detected more frequently in tissue sections. No correlations between markers studied and histological subtypes and grade of carcinoma were found. Taking into account the relationship between P53, BCL-2 and BAX expression, it was possible to classify the studied ovarian carcinomas into four phenotypes. Ovarian cancers with phenotype P53+/BCL-2- were more frequent in III/IV degree FIGO stages whereas the phenotype P53-/BCL-2+ were identified mainly in patients in I/II degree FIGO stages. The progression of disease and death occurred more frequently in groups with phenotype P53+/BCL-2+ and P53-/BCL-2-. CONCLUSIONS: Our observations indicated that estimation of P53, BCL-2 and BAX protein expression may be important in the choice of the chemotherapy.

Assessment of the quality and frequency of mutations occurrence in PTEN gene in endometrial carcinomas and hyperplasias.

The quality and frequency of mutations in PTEN gene were assessed in 59 carcinomas and 6 hyperplasias of the endometrium in women. Screening for mutations was done in all exons of PTEN gene by the PCR-SSCP analysis and DNA sequencing. Results were correlated with histological status and clinical features of endometrial carcinomas. In 45.8% (27/59) of carcinomas, 36 somatic mutations were detected in PTEN gene. In seven carcinomas, two mutations and in one carcinoma three mutations coexisted simultaneously. Moreover in 33.3% (2/6) of hyperplasia cases mutations were shown. Most identified mutations (57.9%) were present in exons 5 and 8, less frequently in exons 2 (15.8%) and 7 (13.2%) and they were least frequent in exons 1 and 3 (5.3% each). No mutations were found in exons 4, 6 and 9. Of all identified mutations, 73.7% of those resulting in truncated protein were present due to deletions, insertions and nonsense mutations. Missense mutations accounted for 13.2% of mutations and they were present only in exon 5. One point mutation (2.5%) was in intronic splice site. The remaining 10.5% of mutations were neutral polymorphisms. No statistically significant correlation were found between the frequency of PTEN gene mutations and the clinical stage of endometrial carcinomas. However, evident statistically significant, reverse correlation were observed between the frequency of mutations and the grade of morphological differentiation of the diseases (chi(2)=7.2393, alpha=0.0071). In conclusion, our data support the view that PTEN gene mutations are frequent events involved in development of endometrial carcinomas in women.

[Own experiences in salvage chemotherapy of ovarian cancer--a preliminary report]. / Doswiadczenia wlasne w ratunkowej chemioterapii raka jajnika. Doniesienie wstepne.

OBJECTIVE: To compare the efficacy and side effects of the ovarian cancer salvage treatment. MATERIALS AND METHODS: 28 patients from 2nd Department of Gynecology of Wroclaw Medical University, treated for recurrent ovarian cancer in stage III and IV were analyzed. First line treatment after complete debulking operation was based on platinum analogue (CAP). Platinum-sensitive patients were administered carboplatin with paclitaxel in the second line treatment. Platinum-resistant cases were treated with paclitaxel, topotecan or liposomal doxorubicin only. RESULTS: The effectiveness of the salvage chemotherapy was similar. Side effects depended on used agent. CONCLUSIONS: We conclude that the results of salvage treatment in ovarian cancer remain unsatisfactory. Introducing new agents into the therapy slightly improved the overall survival time.