Microvesicles released from ectopic endometrial foci as a potential biomarker of endometriosis.

OBJECTIVES: Angiogenesis is engaged in endometriosis. It is regulated by regulatory factors and cytokines, transported in microvesicles. The purpose was to investigate the presence of MVs with vascular endothelial growth factor (VEGF) and metalloproteinase-9 (MMP-9) in peripheral blood and peritoneal fluid of women operated on for endometrioma or teratoma Material and methods:Microvesicles (MVs) were determined in blood samples and peritoneal fluid samples collected from women aged 20-60 years operated on for endometriosis (test group) and teratoma (control group). The final investigations were performed on 47 patients, who qualified for the study based on the meticulous inclusion criteria. MVs were analyzed by flow cytometry (FACS) using annexin V, antibodies for molecules characteristic of cells from endometriosis foci (keratin 18 (K18), CD105, CD146), and antibodies for intraepithelial vascular growth factor VEGF and metalloproteinase-9 (MMP-9). The sample was double "reading" using flow cytometry (FACSCantoII). RESULTS: Cytometry analysis confirmed MVs' presence in plasma and peritoneal fluid collected from patients with both endometriosis and teratomas. A statistically significant higher level of AnnexinV (+) MVs were observed in plasma samples of endometriosis patients. In the control group, there was a higher percentage of double-positive VEGF (+)/MMP-9 (+) and single MMP-9 (+) positive MVs in the serum. In the peritoneal fluid higher frequency of double-positive VEGF (+)/MMP-9 (+) MVs were found in the control group. However, the amount of VEGF (+) / MMP-9 (+) MVs object did not enable to differentiate between the test and control groups. The study was the first, in which MVs were confirmed in plasma and peritoneal fluid in benign adnexa tumors. CONCLUSIONS: Microvesicles are present in peripheral blood and peritoneal fluid samples collected from patients with endometriosis and teratomas. Microvesicles with proangiogenic factors (VEGF and MMP-9) are more abundant in blood and peritoneal fluid samples from patients with teratomas.

Is serum anti-Müllerian hormone (AMH) assay a satisfactory measure for ovarian reserve estimation? A comparison of serum and peritoneal fluid AMH levels.

BACKGROUND: Clinical cases have been reported with women who got pregnant with confirmed low serum anti-Müllerian hormone (AMH) concentrations, thus demonstrating that low serum AMH concentration cut-points could be fairly specific for poor ovarian response (POR) to gonadotrophin stimulation, but not for pregnancy. That observation prompted the question whether serum AMH concentration accurately corresponded to the whole amount of AMH secreted by granulosa cells. OBJECTIVES: To measure AMH levels in peritoneal fluid and their correlations with serum AMH concentrations. MATERIAL AND METHODS: The reported study involved 48 female patients, aged 18-40 years, diagnosed with benign ovarian cysts and qualified for a laparoscopic cystectomy. Prior to surgery, the ovarian reserve was assessed using serum AMH concentration assay. The peritoneal fluid was also collected during the laparoscopy and AMH concentrations in peritoneal fluid were measured. RESULTS: The AMH present in the peritoneal fluid strongly correlated with AMH levels in blood serum (r = 0.54; p < 0.001) and higher serum AMH concentrations corresponded to higher AMH concentrations in the peritoneal fluid. There was also a significant correlation between AMH levels in serum and in peritoneal fluid, collected from patients with endometrioma and other benign cysts (r = 0.61; p = 0.001 vs r = 0.43; p = 0.03). CONCLUSIONS: The AMH is present in the peritoneal fluid and its concentrations significantly correlate with AMH levels in serum. The assessment of AMH concentration in the peritoneal fluid may be a valuable complement to the evaluation of ovarian reserve and the diagnosis of infertility after adnexal surgery.

The significance of markers in the diagnosis of endometrial cancer.

Endometrial cancer is one of the most common cancers experienced by women throughout the world. It is also the most common malignancy within the female reproductive system, representing 37.7% of all disorders. The incidence increases with age, and is diagnosed most frequently in women between 45 and 65 years old. In the last few years, numerous studies have been performed to identify tumour biomarkers. Biomarkers include not only protein routinely used as tumour markers but also genes and chromosomes. The limiting factor in the use of markers in the diagnosis of endometrial cancer is their lack of specificity. However, specific markers for endometrial cancer are the subject of much research attention. Although moderately elevated levels of markers are present in a number of inflammatory or non-malignant diseases, significantly increased levels of markers indicate the development of cancer. Recently, research has been focused on the identification of molecular changes leading to different histological subtypes of endometrial cancer. In this paper the authors reviewed several currently investigated markers. Progress in these investigations is very important in the diagnostics and treatment of endometrial cancer. In particular, the identification of novel mutations and molecular profiles should enhance our ability to personalise adjuvant treatment with genome-guided targeted therapy.

2D/3D ultrasonography for endometrial evaluation in a cohort of 118 postmenopausal women with abnormal uterine bleedings.

OBJECTIVES: 2D/3D transvaginal ultrasonography in evaluation of endometrium in postmenopausal women with abnormal uterine bleedings (AUB). MATERIAL AND METHODS: 2D/3D transvaginal ultrasonography (TVU) was performed in 118 menopausal women with AUB. Endometrial volume and thickness, uterine volume and endometrial vascularity were evaluated. Complete histologic evaluation of the endometrium was obtained through dilatation & curettage (D&C) and/or hysteroscopy. Accordingly, patients were divided into 3 groups: controls (no endometrial pathology, n = 49), GI (benign endometrial pathology, n = 37), GII (endometrial carcinoma, n = 32). RESULTS: GII had greater thickness and volume of the endometrium, compared to GI and controls. The presence of arterial vascular flow was identified only in GI and GII (51.35% and 93.75%, respectively). Endometrial volume merged together with uterine volume measurements (TVU-3D) showed a strong, statistical significance between GI and GII, allowing differentiation of begin and malignant endometrial pathologies in postmenopausal women. CONCLUSIONS: In TVU diagnostics of postmenopausal women with AUB the following play the most significant role: 1) endometrial thickness (TVU-2D); 2) endometrial volume (TVU-3D); 3) uterine plus endometrial volume (TVU-3D); 4) vascularization within the endometrium, allowing to differentiate between pathological and normal endometrium (TVU-2/3D). Evaluation of the endometrial vascularity, both in TVU-2D and TVU-3D technique, does not allow for reliable differentiation between benign lesions and endometrial cancer.

Endometrial and cervical cancer: incidence and mortality among women in the Lodz region.

INTRODUCTION: By the early 21(st) century the most common cancer of female genitals in Poland was cervical cancer. Now endometrial cancer ranks first. The aim of this study was to analyse the incidence and mortality of endometrial and cervical cancer among women in the Lodz region. MATERIAL AND METHODS: Data on the incidence and mortality of endometrial and cervical cancer among inhabitants of the Lodz region were obtained from the National Cancer Registry and Bulletin of Cancer Cases in the Lodz region. The analysis covered ten consecutive years beginning in 2001. RESULTS: The number of new cases reported in 2010 exceeded that observed in 2001 by 181. The standardized incidence rate of endometrial cancer increased by 6.3, while the standardized incidence rate of cervical cancer decreased by 1.4. CONCLUSIONS: In the years 2001-2010, the incidence of endometrial cancer increased by 88.3% and that of cervical cancer decreased by 6.5% among inhabitants of the Lodz region. In the years 2001-2010, mortality of endometrial cancer increased by 24.5% and that of cervical cancer decreased by 12.6%. In 2010, the highest crude incidence rates in the Lodz region of both endometrial and cervical cancer at 39.1 were recorded in the district town of Piotrków.

The toxic impact of local anaesthetics in menopausal women: causes, prevention and treatment after local anaesthetic overdose. Local anaesthetic systemic toxicity syndrome.

Gynaecologists often use local anaesthetics in their medical practice. Some concomitant diseases during the menopausal period may cause problems during the qualification of postmenopausal women for general anaesthesia in gynaecological surgery. Many authors suggest the application of local analgesia for particular kinds of gynaecological surgery procedures performed on postmenopausal women, taking into consideration health determinants. While applying local anaesthetics, the possibility of their overdose has to be taken into account. Generalised toxic symptoms which appeared after the local anaesthesia are rare, but potentially are lethal complications. Toxic symptoms after local anaesthetic administration are manifested after accidental administration of a medicine into a blood vessel, when extravascular administration of a large volume of a local anaesthetic is absorbed into a bloodstream or with the reproducible doses of local anaesthetics which are administered when metabolism does not work sufficiently and cannot eliminate these substances. Clinical overdose of local anaesthetics is manifested by disorders in two systems. Firstly, the pathological symptoms come from the central nervous system (CNS). In the second phase, the pathological symptoms will additionally appear in the cardiovascular system. The aim of the present thesis is to remind clinical manifestations of the local anaesthetic overdose and suggest the management of patients with the aforementioned symptoms, especially in the case of intravenous lipid emulsions which have the status of an antidote in life-threatening conditions caused by cardiotoxic effects of local anaesthetics.

Hemostatic disorders of the menopausal period: the role of microRNA.

Adverse changes in hemostasis of menopausal women, observed e.g. in atherosclerotic or neoplastic cases, are of multicausal origin. It is believed that in the development and regulation of these processes, an important role is played by microRNA particles, which presence is ascertained in endothelial cells, atherosclerotic plaques and systemic circulation. Discovered for the first time over 20 years ago, up to now over two and a half thousand types of microRNA have been identified in the human body. MicroRNAs are single stranded RNA molecules of 20-24 nucleotides, encoded by the cell's genome and then transcribed by polymerase II. They regulate the expression of a large gene pool, approximately 30% of all genes, in the human body. MicroRNA molecules, like other bioactive molecules - RNA, protein - both play important roles in tumor invasion, metastasis, inflammation, coagulation, and regeneration. What is important, they can be detected not only in tissues (e.g. tumor tissues), but also in circulation (blood serum), where they are released. Accurate understanding of the role played by certain types of microRNA (e.g. miR-126, miR-17-92, miR-33, miR-613, miR-27a/b, miR-143, miR-335, miR-370, miR-122, miR-19b, miR-520, or miR-220) in hemostatic processes may allow in the future for their use not only as specific biomarkers of cardiovascular diseases but also as the target for innovative gene therapies.

Current clinical application of serum biomarkers to detect ovarian cancer.

For the last decades, hundreds of potential serum biomarkers have been assessed in diagnosing of ovarian cancer including the wide spectrum of cytokines, growth factors, adhesion molecules, proteases, hormones, coagulation factors, acute phase reactants, and apoptosis factors but except CA125 none of them have been applied to everyday clinical practice. Nowadays, the growing number of evidence suggests that the classic marker CA125 should be accompanied by HE4 and in fact, Risk of Ovarian Malignancy Algorithm (ROMA) is becoming more and more widespread in clinical practice for the evaluation of adnexal masses. Early ovarian cancer is often asymptomatic, so the challenge still exists to develop serum markers suitable for early diagnosis and screening. Current knowledge strongly points to different mechanisms of pathogenesis, genetic disturbances and clinical course of major histological subtypes of ovarian cancer. Thus, future biomarker/multimarker panels should take into consideration the implications of different molecular patterns and biological behavior of various subtypes of ovarian cancer. Very promising are studies on miRNAs - small non-protein coding gene-regulatory RNA molecules functionally involved in the pathogenesis of cancers acting as oncogenes (oncomirs) or tumor suppressors. The studies devoted to ovarian cancer tissue miRNA profiling have shown that miRNAs could be useful in diagnosing and predicting the OC outcome. They also confirmed that OC is a highly heterogeneous disease, gathering four distinct histological tumor subtypes characterized not only by distinct origin, behavior and response to chemotherapy but also by different patterns of miRNA expression.

Microvesicles as a potential biomarker of neoplastic diseases and their role in development and progression of neoplasm.

Neoplastic diseases together with cardiovascular diseases are the most frequent causes of death in the Polish population. Cancers of reproductive organs with breast cancer are responsible for the highest morbidity and mortality in women suffering from neoplasm diseases. Asymptomatic dynamics of the development of a neoplasm and no deviations from the normal level of laboratory results contribute to the fact that malignant diseases are diagnosed too late. The aim of modern medicine is to diagnose cancer at the earliest stage, however, there is no sufficiently sensitive and specific biomarker which can be used for diagnostic, prognostic and therapeutic purposes. Cellular interactions play the main role in the development, angiogenesis and invasiveness of a tumor. Recent research suggests the possibility of microvesicles (MVs) involvement in communication between cells. The MVs ability to fuse with various cells is used in cell-to-cell contact. Microvesicles cargo may include growth factors, their receptors, protease, adhesion molecules, signaling molecules and the sequence of DNA, mRNA, and micro-RNA. Larger quantities of MVs released from neoplastic cells affect both the local environment and systematic range causing metastases and progression. The research on molecular mechanisms of MVs' release and the presence of characteristic oncogenes in blood of patients with neoplasms is being carried out. Confirmation of MVs presence in patients' serum can potentially serve as useful information for therapeutic purposes and as the biomarker of a neoplastic disease.

Leiomyomatosis peritonealis disseminata of unusual course with malignant transformation: case report.

Leiomyomatosis peritonealis disseminata is a very rare, benign entity of unknown pathogenesis, characterized by the presence of multiple subperitoneal or peritoneal smooth muscle nodules throughout the peritoneal surface. Mostly the course is asymptomatic and it is found incidentally during laparotomy, laparoscopy or cesarean section. Non-specific symptoms such as abdominal pain, vaginal bleeding, abdominal mass or gastrointestinal signs are described. Rare cases of malignant transformation have been reported. We present a case of disseminated peritoneal leiomyomatosis with an unusual course and transformation to endometrial sarcoma in a 26-year-old previously healthy woman, where the appearance of peritoneal nodules was preceded by multiple incidents of fast fibroid growth and delivery of myomatous growth into the cervical canal.

The use of panniculectomy in morbidly obese patients undergoing gynecological surgery.

Panniculectomy is a surgical procedure that involves removal of the skin and fat excess which facilitates access to the peritoneal cavity. In the paper we present three cases of morbidly obese women (BMI: 46.3-59.5) who were treated in the Department of Gynecology and Oncological Gynecology in Lodz. One of the patients underwent an operation due to the presence of a large cervical myoma. Two another women were treated for endometrial cancer. During all of the three procedures panniculectomy was the first stage of the operation.

The role of angiogenic factors in endometrial cancer.

Endometrial cancer is the most common malignancy within the female reproductive system (37.7%). The incidence increases with age. Frequently this type of cancer is diagnosed in peri- and post-menopausal women. 60-70% of cancers occur in women over 60 years of age, and less than 5% in women below 40 years of age. Angiogenesis is a process of formation of new microvessels from existing capillaries. There are four different mechanisms of new vessel growth: sprouting, intussusception, vessel elongation and incorporation of endothelial progenitor cells into new microvessels. Angiogenesis plays important roles in growth of endometrial cancers. This process is controlled by many angiogenic factors, for example vascular endothelial growth factor (VEGF). VEGF is the most powerful and most specific endothelial cell growth factor. It plays a crucial role in the initiation of physiological and pathological angiogenesis, lymphangiogenesis, and vasculogenesis. The VEGF family consists of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F and PLGF (placental growth factor). The effects of VEGF are mediated through binding to the two specific and homologous receptors VEGFR-1 (FLT-1) and VEGFR-2 (KDR). Placental growth factor (PLGF) belongs to the VEGF family and it is also a very important growth factor. So far four isoforms of PLGF have been identified: PLGF-1 (PLGF131), PLGF-2 (PLGF152), PLGF-3 (PLGF203) and PLGF-4 (PLGF224).

Incidence of and mortality from breast cancer among women in Poland in the years 2001-2010.

INTRODUCTION: Breast cancer is the most common malignant tumour among women. About 15,000 new cases of breast cancer are diagnosed and more than 5,000 women die in Poland every year. The aim of this study was to analyse the incidence and mortality rate of breast cancer among women in Poland in the years 2001-2010. MATERIAL AND METHODS: Analysed data concerning the incidence of and mortality from cancer among women were obtained from the National Cancer Registry. RESULTS: The number of new cases reported in 2010 exceeded that reported in 2001 by 3,666. The mortality from breast cancer among women increased by 15.1% by 2009, to subsequently drop by 0.3% in 2010. The standardized incidence rate increased by 7.4 and the standardized mortality rate fell by 1.3 in 2001-2010. CONCLUSIONS: In the years 2001-2010 the incidence of breast cancer in women in Poland rose by 30.3%, with an increase of 7.4 in the incidence rate. The highest rise in the incidence and mortality of women due to breast cancer in Poland is reported in the Lodz voivodeship. In the years 2001-2009 the number of women's deaths due to breast cancer increased slightly, while the mortality rate dropped.

Polymorphism of the ERalpha and CYP1B1 genes in endometrial cancer in a Polish subpopulation.

AIM: Metabolic activation of estrogens may play a role in endometrial carcinogenesis; and polymorphism of the genes (whose product is involved in this process) may be associated with the modulation of the risk of endometrial cancer. CYP1B1 plays a major role in the metabolism of estrogens, which must firstly bind their receptors, estrogen receptor alpha (ERalpha) or ER beta. In the present study we investigated the association of two polymorphisms of the CYP1B1 gene (Arg48Gly [142C > G] and Leu432Val [4326C > G]) and a polymorphism of the ERalpha gene (975C > G) as well as a combination between them with endometrial cancer occurrence. METHODS: Genotypes were determined in DNA from peripheral blood lymphocytes of 100 endometrial cancer patients and 100 age- and ethnically-matched cancer-free controls by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). RESULTS: We found an association between endometrial cancer occurrence and the Arg48Arg (142C > C)-CYP1B1 variant (odds ratio [OR] 2.10; 95% confidence interval [CI] 1.17-3.79) and the 975C > G -ERalpha gene polymorphism (OR 3.84; 95%CI 2.08-7.10). Gene-gene interaction between the Arg48Arg (142C > C) and Leu432Val (4326C > G) variants and between Arg48Gly (142C > G) -CYP1B1 and 975C > G -ERalpha as well as Gly48Gly (142G > G)-CYP1B1 and 975C > G-ERalpha increased the risk of endometrial cancer (OR 2.70; 95%CI 1.12-6.49; OR 2.52; 95%CI 1.04-6.11 and OR 3.62; 95%CI 1.27-10.30, respectively). Additionally interaction between the 975C > G-ERalpha and Leu432Leu (4326C > C)-CYP1B1 or Leu432Val (4326C > G)-CYP1B1 variants also increased the risk (OR 4.68; 95%CI 1.81-12.07 and OR 6.00; 95%CI 2.19-16.47, respectively). CONCLUSIONS: The CYP1B1 and ERalpha genes may play a role in endometrial cancer and the Arg48Gly (142C > G) -CYP1B1 and 975C > G-ERalpha polymorphisms may be considered as independent, early diagnostic markers in this disease.

DNA damage and repair in children with Down's syndrome.

Down's syndrome (DS) is associated with the presence of a third 21 chromosome and is generally considered as a non-cancer-prone genetic disease. However, leukaemias occur more frequently in children with the syndrome than in general population and there is an open question, whether the presence of an additional chromosome may contribute to genomic instability, which, in turn, may play a role in a higher susceptibility to cancer and leukaemias in particular. In order to assess genomic instability associated with the presence of a third 21 chromosome, we determined the level of endogenous DNA damage and susceptibility to a genotoxic stress-inducing factor, hydrogen peroxide and N-methyl-N'-nitro-N-nitrosoguanidyne (MNNG) as well as the ability to remove DNA damage in the peripheral blood lymphocytes of children with DS and healthy kids. The level of DNA damage and the kinetics of DNA repair were evaluated by alkaline comet assay. Oxidative DNA damage was assayed with DNA repair enzymes: endonuclease III-like NTH1 and formamidopyrimidine-DNA glycosylase. The cells taken from children with DS did not display an effective DNA repair after treatment with 10 mM hydrogen peroxide. No difference in the sensitivity to DNA-damaging agents and the efficacy of DNA repair due to age and gender in DS children was observed. These results suggest that children with DS may be characterized by the increased sensitivity to the DNA-damaging agents impaired cellular reaction to DNA damage, which, in turn, may increase the probability of cancers in these children. Therefore, a special care to avoid exposure to potential mutagenic factor my be considered in these children.

The role of BRCA1 gene mutations and apoptosis phenomenon in sporadic breast cancer.

BRCA1 tumor suppressor gene encodes an 1863-amino acid gene product that is implicated in many cellular pathways including transcription, cell-cycle checkpoint control, apoptosis and DNA repair. A role of apoptosis and BRCA1 germ-line mutation in breast cancer appearance was investigated in this study by both apoptosis frequency analysis and mutation screening of BRCA1 among breast cancer cases. Blood was obtained from 40 women with node-negative and node-positive ductal breast carcinomas with uniform tumor size. The blood samples from age matched healthy women (n=42) served as control. BRCA1 gene mutations were determined by PCR-RFLP methods. The apoptotic peripheral blood cells were detected by agarose gel electrophoresis. The apoptotic cells were identified in 30% (12/40) of the patients. There were no significant differences in apoptosis frequencies between patients and controls (P > 0.05). Three mutations of BRCA1 gene were identified in apoptosis positive samples from breast cancer women; one Ex20insC and two ExII17delA. Our study implies that apoptosis may be involved not only in sporadic breast carcinoma without BRCA1 mutations, but also in BRCA1-associated breast carcinoma.

[Evaluation of endometrial biopsy in postmenopausal women before HRT]. / Przydatnosc biopsji aspiracyjnej endometrium u kobiet po menopauzie, kwalifikowanych do hormonalnej terapii zastepczej.

UNLABELLED: One of the reasons for not commencing or withdrawal of HRT in women is their fear for breast and endometrial cancers. Does ultrasonographic valuation guarantee sufficiently patient's safety? Most investigators do not recommend further endometrial diagnostics with endometrial thickness less than 4 mm. Endometrial biopsy is advised in cases of irregular uterine bleedings with normal endometrial image or when any endometrial pathology is suspected. In Poland the most frequently performed procedure of endometrial diagnostics is D&C. Our proposition was to offer endometrial biopsies to all patients who were taken HRT for more than 5 years or, despite having strong climacteric ailments, refused to take it because of their cancerophobia. AIM OF STUDY: To evaluate the usefulness of aspiration endometrial biopsy in women after menopause qualified for HRT. MATERIAL AND METHODS: In 84 females, being postmenopausal (amenorrhoeic for at least 12 months) and qualified for HRT, aspiration endometrial biopsies were performed in outpatient clinics. Age of women ranged 46-63 years, mean 57.3. Attained results were compared to ultrasonographic endometrial evaluation. Such factors as patient's age, menopausal age (years from last menstrual period) and previous administration of HRT were taken into our account (maintaining a 3-month wash-out interval). In 74% of women we have got a material being sufficient for histological evaluation. Form clinical point of view the most important for us was to exclude a proliferative or neoplastic process within endometrium. One of endometrial polyps was omitted in USG study, we were surprised also while detecting endometrial carcinoma cells in endometrium below 3.5 mm. In one postoperative slide the cells of endometrial ovarian carcinoma (with infiltrated Fallopian tube) were detected. CONCLUSION: Endometrial biopsy seems to be a useful, effective and cheap method of endometrial diagnostics also in women after menopause.

[Evaluation of the effect of obesity on selected haemostatic and fibrinolytic parameters in peri- and postmenopausal women]. / Ocena wplywu otylosci na wybrane parametry ukladu krzepniecia i fibrynolizy u kobiet w okresie okolo-i pomenopauzalnym.

Localization of adipose tissue influence on obesity-dependent diseases. In obese people many abnormalities are observed connected with coagulation factors and fibrinolysis. We studied 47 peri- and postmenopausal female patients, who were operated on for benign changes within the pelvis minor. The patients were divided according to BMI (group I--BMI < 25 kg/m2, group II--BMI > 29 kg/m2). In obese women lower SHBG concentration, higher insulin and glucose fasting blood levels, higher LDL, higher total fatty tissue content, higher fibrinogen level and elevated PAI-1 activity were found. A positive correlation between area under insulin curve and PAI-1 activity and a negative correlation between fasting insulin levels and tPA were also observed. Based on the studies performed it can be said that: 1. Fatty tissue content in postmenopausal women exerts an influence on PAI-1 activity. 2. Obesity influences coagulation and fibrinolysis system in peri- and postmenopausal women. 3. Carbohydrate metabolism disturbances have influence on PAI-1 activity in peri- and postmenopausal women.