Regulatory T lymphocytes and transforming growth factor beta in epithelial ovarian tumors-prognostic significance.

BACKGROUND: Regulatory T lymphocytes (Treg) are characterized by the presence of CD4+ surface antigen. Today the transcription factor FOXP3 is considered to be the most specific marker of Treg cells. The aim of the study was to estimate the percentage of Treg in peripheral blood and the tissue of the epithelial ovarian tumor and blood serum TGF-beta concentrations and relationships between them. Moreover, the aim of the study was to answer the question whether the percentage of Treg lymphocytes affects the time of survival in patients with ovarian cancer. METHODS: The patients were divided into four groups, depending on the histopathological examination result: I--a group without any pathology within the ovaries (C; n = 20), II--a group with benign tumors (B; n = 25), III - with borderline tumors (BR; n = 11), IV--a group with cancer of the ovary (M; n = 24). The percentage of Treg lymphocytes in peripheral blood and the tissue was assessed using the flow cytometry method. TGF-beta cytokine concentration was estimated with the ELISA immunoenzymatic test. Statistical analysis of the results was conducted using the computer program Statistica 10.0PL (StatSoft, Inc). RESULTS: No significant differences were found in percentages of Treg lymphocytes in peripheral blood between individual groups of patients (p = 0.11). However, we observed marked differences in the tissue of malignant and non-malignant tumors between individual groups of patients (p = 0.003). The analysis with the post hoc test revealed significantly higher TGF-beta concentration in the group of women with malignant tumors. Moreover, no relationship was found between TGF-beta concentration and the percentage of Treg cells in peripheral blood and tumors of the ovary. No correlation was found between the percentage of Treg lymphocytes in peripheral blood (p = 0.4) and the tissue of ovarian tumors (p = 0.3) and the time of survival of patients with ovarian cancer. CONCLUSIONS: The recruitment of Treg lymphocytes toward the tumor is one of the mechanisms of escape of neoplasm from the response of the immune system. The percentage of Treg lymphocytes in peripheral blood and the neoplastic tissue does not influence the time of survival of patients with ovarian cancer.

8-oxo-7,8-dihydroguanine level - the DNA oxidative stress marker - recognized by fluorescence image analysis in sporadic uterine adenocarcinomas in women.

OBJECTIVES: In the case of carcinogenesis in human endometrium no information exists on tissue concentration of 8-oxo-7,8-dihydroguanine, the DNA oxidative stress marker This was the main reason to undertake the investigation of this DNA modification in human uterine estrogen-dependent tissue cancers. MATERIAL AND METHODS: In order to estimate the level of oxidative damage, 8-oxo-7,8-dihydroguanine was determined directly in cells of tissue microscope slides using OxyDNA Assay Kit, Fluorometric. Cells were investigated under confocal microscope. Images of individual cells were captured by computer-interfaced digital photography and analyzed for fluorescence intensities (continuous inverted 8-bit gray-scale = 0 [black]-255 [white]). Fluorescence scores were calculated for each of 13 normal endometrial samples and 31 uterine adenocarcinoma specimens. Finally the level of the oxidative stress marker was also analyzed according to histological and clinical features of the neoplasms. RESULTS: The obtained data revealed that: 8-oxo-7,8-dihydroguanine levels were higher in uterine adenocarcinomas than in normal endometrial samples (48,32 vs. 38,64; p<0,001); in contrast to normal endometrium there was no correlation between age and DNA oxidative modification content in uterine cancer; highest mean fluorescence intensity was recognized in G2 endometrial adenocarcinomas; level of 8-oxo-7,8-dihydroguanine does not depend on Body Mass Index (BMI) and cancer uterine wall infiltration or tumor FIGO stage. CONCLUSIONS: Our study indicates that accumulation of the oxidized DNA base may contribute to the development of endometrial neoplasia, however oxidative DNA damage does not seem to increase with tumor progression.

PARP-1 activity in normal and cancerous human endometrium and its relationship with quantity of abasic sites (AP).

OBJECTIVES: Poly (ADP-ribose) polymerase (PARP-1) is involved in the processes of DNA repair contributing to the maintenance of genomic stability. Recent data suggest that polymerase is involved in the development of endometrial adenocarcinomas and more advanced tumors displaying lowest enzyme protein expression. Data on PARP-1 activity regarding carcinogenesis in human endometrium are scarce. That was the reason why the authors of the present work wished to investigate the enzyme activity in human uterine hormone-dependent cancer and to compare the results with those obtained for normal endometrial tissue. The next aim was to check whether enzyme activity in normal and cancerous endometrium depends on the number of AP sites, which are widely known as oxidative stress DNA damage markers and PARP-1 activity stimulators. MATERIAL AND METHODS: Universal Colorimetric PARP Assay Kit was used to estimate the enzyme activity in units/ mg protein. Apurinic sites/105 base pairs (bp) were measured by Oxidative DNA Damage Kit Quantitative. Results were calculated for 47 endometrial samples and 15 uterine adenocarcinomas specimens. Finally the PARP-1 activity was analyzed for histological and some clinical features of neoplasms. RESULTS AND CONCLUSIONS: 1. no differences in PARP-1 activity were found in non-cancerous types of human endometrium; 2. mean enzyme activity was lower in sporadic endometrial cancers than in noncancerous endometrial specimens (2.89 +/- 0.55 vs 6.39 +/- 0.06; p < 0.005); 3. mean PARP-1 activity in lower grade neoplasms was higher than in G3 tumors and was lower in adenocarcinomas displaying deep uterine wall infiltration; 4. there was no relationship between PARP-1 activity and AP level.

Estrogen and progesterone receptor expression in HPV-positive and HPV-negative cervical carcinomas.

Human papillomavirus (HPV) is widely accepted as the main cause of cervical cancer. However, the presence of HPV DNA does not inescapably lead to the development of the cancerous phenotype of the infected cell. Therefore, it is considered that the induction of full cancerous expression of HPV requires additional cofactors. The aim of this study was to assess the expression of estrogen receptor α (ERα) and progesterone receptor (PR) in archived tissue blocks of squamous cell carcinoma and adenocarcinoma of the uterine cervix and to ascertain whether expression of these receptors is associated with the presence of HPV DNA. The investigation was performed using formalin-fixed, paraffin-embedded cervical cancer specimens obtained from 250 women who underwent surgery for histologically confirmed neoplastic lesions. The control group consisted of normal cervical tissues obtained from 50 patients who underwent myomectomy. The results of this study revealed that the expression of ER and PR in planoepithelial cancers and adenocarcinomas of the cervix were decreased to undetectable levels. Only in singular cases in the pattern of staining the expression of ER and PR was noted. In stromal cells of the tested neoplasms, higher expression of both types of receptors was found. Comparison of the expression of ER and PR in the staining pattern and stroma of both squamous cell carcinoma and adenocarcioma of the cervix, showed statistically higher expression in the stromal cells. Strong expression (+1, +2, +3) of ER and PR was noted in the stromal cells irrespective of HPV infection, histopathological type of cancer, and clinical and histopathological grade.

[Vesicovaginal fistulas--current therapeutical strategy]. / Przetoki pecherzowo-pochwowe--aktualna strategia postepowania.

OBJECTIVES: Clinical evaluation of Latzko operation in primary and recurrent vesicovaginal fistulas and an update on current therapeutical strategy. MATERIAL AND METHODS: Evaluation of clinical efficacy has been performed in a group of 5 patients with confirmed vesicovaginal fistulas and in 1 woman with urethrovaginal fistula. Prior to the operation, four patients had undergone eight fistula operations in our department with unsuccessful outcome. RESULTS: Fistula closure by Latzko operation has been achieved in 5 patients, including one patient who had surgery treatment repeated 3 months after the primary fistula operation. In total, additional Martius flap has been introduced twice. In patient, where fistula has been operated after 45 days since primary TVM and IVSO4 surgery closure therapy failed. CONCLUSIONS: Surgical treatment by vaginal approach offers great therapeutical efficacy in the primary and recurrent vesicovaginal fistulas, even in cases when previous abdominal surgical treatment had failed.

[Apurinic/apyrimidinic sites (AP)--stress oxidative marker--in normal and neoplastic human endometrium]. / Liczba miejsc apurynowych/apirymidynowych (AP)--markera stresu oksydacyjnego--w prawidlowym i nowotworowo zmienionym endometrium u kobiet.

OBJECTIVES: To evaluate apurinic/apyrimidinic sites (AP), one of the oxidative stress markers in normal and cancerous human endometrium and determine whether AP sites could be a molecular marker of endometrial cancer advancement. MATERIAL AND METHODS: AP sites were investigated in DNAs of 33 endometrial cancer (EC) and 20 noncancerous endometrial samples using Oxidative DNA Damage Kit Quantitative (Kamiya Biomedical Company). RESULTS: Mean AP sites in noncancerous endometrium was 6.0 +/- 1.21 per 105 base pair. In EC group the mean AP sites level was greater than estimated in the reference group containing proliferative, secretory and hyperplastic endometrial samples (p < 0.05). There was no relation between AP sites and EC FIGO grading or the depth of the uterine wall neoplastic invasion. CONCLUSIONS: In EC mean AP sites level is greater than estimated in noncancerous endometrium. AP sites are not a molecular marker of EC advancement.

[Conservative treatment of cervical pregnancy in women: a review of the literature with case report]. / Zachowawcze leczenie ciazy szyjkowej u kobiet: opis przypadku i przeglad literatury.

Diagnosis of cervical pregnancy depends on ultrasonographic image of the gestational sac within the cervix, rather than histopathologic examination. Diagnosis of a cervical pregnancy may be confirmed if the placenta and the entire chorionic sac containing a live fetus are located below the internal os. Majority of patients with cervical pregnancies are young women who wish to stay fertile and have the possibility to bear children. Ultrasonography offers the advantage of early clinical diagnosis and easy fallow-up. Methotrexate (MTX) has become the treatment of choice in cervical pregnancy management, especially for the hemodynamically stable patients. In the following article we have reported a the successful management of a 7 weeks gestation cervical pregnancy treated with the combination of MTX, prostaglandin and suction curettage. Due to conservative treatment emergency hysterectomy was avoided.

Is global DNA methylation in sporadic uterine adenocarcinomas in women a result of histological and clinical tumor advancement?

AIM: To find out the relationship, if any between the extent of the overall genomic DNA methylation, and clinical and pathological features of the sporadic endometrial adenocarcinomas in women. MATERIAL: Genomic DNA was isolated from 44 primary uterine cancer tissue specimens. There were eight G1, 24 G2 and twelve G3 tumors. METHODS: m5dC level was estimated after enzymatic digestion of DNA into nucleotides, 32P-postlabelling, two-dimensional thin-layer chromatography on cellulose plates and phosphorobioimaging. The overall m5dC of the uterine cancer DNAs expressed as a ratio: (pm5dC/pm5dC+pdC) x 100% was compared to results obtained for parallel investigated but published earlier normal human endometrium DNAs. RESULTS: Mean total cancer DNA methylation (3.48+/-0.46%) was significantly higher than that of the normal proliferative endometrium (2.94+/-0.4%, p=0.003) and lower than that of the secretory endometrium DNAs (3.75+/-0.47%, p=0.03). Among all endometrial cancer DNAs six were found to be hypomethylated, eight were hypermethylated, whereas the remaining 30 had m5dC within range of normal endometrium. Total DNA methylation was significantly higher in poorly differentiated (G3) than in lower grade neoplasms (3.94+/-0.46 vs. 3.3+/-0.32%, p=0.025). Lower levels of DNA methylation seemed to be associated with diminished tumor invasiveness. CONCLUSIONS: Our results suggest that alterations in overall DNA methylation seem to be a result of neoplastic transformation and could therefore be used as a prognostic molecular marker of endometrial cancer.

Changing trends in the surgical treatment of female stress urinary incontinence--twenty two years observation.

OBJECTIVES: The aim of this study was to analyze the changing trends in surgical treatment of female urinary incontinence (UI). MATERIAL AND METHODS: Medical records of all women admitted to II Department of Gynecology from 1985 to 2006 were analyzed in order to find out how the female SUI treatment changed over these years. RESULTS: During analyzed time 36819 patients were hospitalized in our Department and 77.6% (28568) of them were operated because of various indications. The number of SUI surgeries among all hospitalized women steadily rose from 1.93% in 1985 to 10.96% in 2006 reaching maximum in 2005 (13.73%). Clinical effectiveness of SUI surgeries markedly improved from 35% for anterior colporrhaphy to almost 90 % for suburethral slings. CONCLUSIONS: Introduction into clinical practice modern suburethral slings improved clinical efficacy of SUI treatment. The percentage of women admitted and treated surgically because of SUI steadily increased over the last years.

Activity of matrix metalloproteinase-2 and -9 and contents of their tissue inhibitors in uterine leiomyoma and corresponding myometrium.

BACKGROUND AND AIM: Matrix metalloproteinase-2 and -9 (MMP-2 and -9) are proteolytic enzymes degrading extracellular matrix proteins, mainly collagen type IV. Recent reports show that these proteases may be implicated in the growth of uterine leiomyoma. The aim of the present study was to evaluate the activity of MMP-2 and MMP-9, the contents of their tissue inhibitors (TIMP-1 and TIMP-2) and the immunolocalization of collagen type IV in uterine leiomyoma and corresponding myometrium. MATERIALS AND METHODS: Material for the study comprised specimens of uterine leiomyomas and corresponding myometrium derived from 20 hysterectomized women. The activity of MMP-2 and MMP-9 in tissue extracts was evaluated by semi-quantitative zymography. TIMPs were measured by enzyme-linked inmmunosorbent assay. Protein immunohistochemistry was applied for detection of collagen type IV. RESULTS: Activity and activation ratio of MMP-2 were significantly higher in leiomyomas than myometrium. The activity of MMP-9 was weak and did not differ between the investigated tissues. Contents of TIPM-1 and TIPM-2 were similar in both tissues. In both leiomyomas and myometrium, collagen type IV was localized in the extracellular matrix embedding bundles of smooth muscle cells, but was absent in areas of extracellular matrix accumulation within leiomyomas and in larger septa separating muscle fibers in normal myometrium. CONCLUSION: MMP-2 may be implicated in pathogenesis of leiomyoma.

[DNA adducts in human female genital organs]. / Addukty DNA w narzadach plciowych kobiety.

DNA adducts, one of genetic damages markers, precede and finally can lead to oncogenic mutations. They appear in genome as a result of DNA bases damages caused by various and numerous environmental factors eg. ultraviolet light, ionic radiation, toxins and also endogenic substances, for example estrogens. It is believed that the creation of DNA adducts is a necessary but insufficient process for the neoplastic transformation of the cell. The following review presents concise knowledge about the DNA adducts creation and their sequels served in healthy and cancerous tissues of the female genital organs, on the base of the available data.

Immunohistochemical detection of hMLH1 and hMSH2 proteins in vulvar carcinoma.

The immunohistochemical (IHC) detection of MMR proteins is an accurate and rapid method to predict the presence of defective DNA MMR genes. MMR protein expression could also serve as a prognostic indicator of human cancers. The results of many studies demonstrate the usefulness of IHC tests with monoclonal antibodies MSH2 and MLH1 in screening the microsatellite sequence instability within both spontaneous and hereditary malignant neoplasms. The aim of our study was to perform an IHC estimation of the hMLH1 and hMSH2 expression in a subset of vulvar carcinomas according to HPV 16/18 status. The level of MMR proteins was further analyzed in relation to histoclinical features of the disease in either HPV-positive or -negative cancers. We identified archival diagnostic phase tissue specimens from 46 cases of vulvar cancer. From the same paraffin blocks containing material from the margin of surgical section during vulvectomy, normal epithelial tissue fragments were collected and designated as the control group. The characteristic of the lesion was examined in comparison with the presence of HPV DNA. Identification of the HPV 16/18 types was performed using PCR. IgG1 monoclonal antibodies detecting those epitopes characteristic for hMLH1 and hMSH2 were used in the study. In the analyzed cases of vulvar cancer, we have observed increased expression of proteins of both hMSH2 and hMLH1 genes compared to the control group. A comparison of the hMLH1 and hMSH2 protein expression levels showed that hMSH2 expression was higher than that of hMLH1 in the case of vulvar carcinomas. The performed analysis of correlation between individual parameters did not reveal statistically significant relationship with both the gradient and status of HPV 16/18. hMSH2 and hMLH1 were definitely interrelated.

Global DNA methylation in relation to hMLH1 and hMSH2 protein immunoreactivity in sporadic human endometrial carcinomas.

Overall DNA methylation status was studied in a group of 28 sporadic human endometrial carcinomas (ECs) using the [32P]-postlabeling technique. Moreover, expression of the DNA mismatch repair proteins (hMLH1 and hMSH2) was investigated in ECs using immunohistochemistry. Mean 5-methyldeoxycytosine (m5dC) content in the studied group was 3.48+/-0.37% (range, 2.89-4.12%). The mean m5dC scores were significantly different between early (3.35+/-0.33%) and advanced (3.66+/-0.36%) endometrial neoplasms (chi2-test; p=0.03). There was a markedly increased overall DNA methylation with the degree of histological differentiation and with the infiltration of the myometrium (p<0.05). Loss of hMLH1 and hMSH2 expression was reported in 7 (25%) and 5 (18%) tumors, respectively, but the immunoreactivity did not correlate with the known clinicopathological variables of cancer. In addition, no obvious correlation was found between global m5dC content and the lack of hMLH1 and hMSH2 protein expression in human uterine tumors (p=0.97 and p=0.19 for hMLH1 and hMSH2, respectively; Spearman's rank correlation test). Our results clearly show that alterations in global DNA methylation may influence tumor progression, but they are not directly associated with the inactivation of the mismatch-repair machinery in sporadic human ECs.

[TVT procedure--the critical analysis of clinical effectiveness and complications among first 100 cases]. / Operacja TVT--analiza skutecznosci oraz powiklan pierwszych 100 operacji.

RATIONALE: Since its clinical debut in 1996 TVT procedure has been offering an excellent clinical tool both for patients and surgeons to treat SUI suffering women. Since the learning curve is an important factor influencing the rate of surgical complications as well as the clinical outcome of any new surgical technique we decided to analyse the first 100 cases of SUI women treated with TVT technique. AIM OF THE STUDY: To analyse clinical effectiveness and complications among first 100 consecutive cases of TVT procedures. MATERIAL AND METHODS: One hundred women aged from 30 to 89 years (old mean 55.6) were hospitalised in our Department from September 1999 to November 2000 because of SUI as confirmed by the complete urogynecologic assessment including history and physical examinations, catheterised residual volume determination, and multichannel urodynamic testing. TVT procedure was performed as originally described by Ulmsten et al. except the fact that 43 operations were performed not under local but epidural anesthesia. For 80 patients TVT was offered as the primary surgery for SUI treatment whereas for 14 it was the second and for 6 the third or more attempt. RESULTS: Operation was performed without any technical difficulties in all cases (mean time 25 min). Bladder injury (treated by 1 or 2 days catheterization) occurred in 9 cases, urinary retention in 13 (in 6 cases after 10-14 days tape was cut). Urinary tract infections occurred in 4 women despite intraoperative 2.0 g i.v. piperacillin prophylactics and fosfomycin trometamol 3.0 g p.o. in first postoperative day. Subsequent urge incontinence developed in 4 patients. Clinical efficacy based on medical history, cough test and Gaudenz questionnaire was 95% (observation period 3 to 16 months, mean 8 months). CONCLUSIONS: In our opinion TVT procedure is an operation of choice to treat SUI in women regardless of their age because of its excellent clinical effectiveness accompanied by its technical simplicity, reproducibility and low percentage of serious complications. Previous urogenital surgery is a risk factor for bladder perforation during TVT procedure.

Serum concentrations of markers of type I collagen metabolism in women taking monophasic oral contraceptives.

OBJECTIVES: To investigate serum concentrations of the carboxy-terminal propeptide of type I procollagen (PICP) and cross-linked carboxy-terminal telopeptide of type I collagen (ICTP), reflecting the rate of synthesis and degradation of the parent collagen respectively, in women using monophasic oral contraceptives. STUDY DESIGN: PICP and ICTP were estimated in 60 women taking 20 micrograms of ethinyl estradiol (EE) + 150 micrograms of desogestrel (DSG) or 30 micrograms of EE + 150 micrograms DSG over six months. RESULTS: Mean PICP concentration decreased in women receiving 20 micrograms of EE + 150 infinity g of DSG after three cycles of administration. However, after six months this value returned to a level comparable to the initial one. In women taking 30 micrograms of EE + 150 micrograms of DSG the mean concentrations of PICP did not change significantly throughout the period studied. None of the oral contraceptives noticeably influenced the concentrations of ICTP. CONCLUSION: Investigated oral contraceptives do not significantly affect the metabolism of type I collagen, however a transient decrease in its biosynthesis may be expected during the use of formulations containing 20 micrograms of EE.

Is diminished pubocervical fascia collagen content a risk factor for failure of surgical management of genuine stress urinary incontinence in women?

OBJECTIVE: The assessment of relationship between pubocervical collagen content and clinical results of surgical treatment of genuine stress urinary incontinence (GSUI) in women. METHODS: Twenty-four women treated for genuine stress urinary incontinence were included into the study. All women underwent the same surgical procedure. The samples of pubocervical fascia were taken at the time of surgery. The contents of acid soluble, pepsin soluble, insoluble fraction of collagen, total collagen and collagen crosslinks were measured. The study of pubocervical fascia collagen metabolism included also estimation of collagenase activity. At follow-up done 5 years following surgery, 20 patients reported symptoms of GSUI (study group). Four women were still without symptoms of urine leakage (control group). RESULTS: The biochemical parameters of pubocervical fascia did not show, statistically significant differences between compared groups. CONCLUSION: The pubocervical fascia collagen metabolism does not have impact on the results of anti-incontinence surgery.