Anti-Apoptotic Effect of Apelin in Human Placenta: Studies on BeWo Cells and Villous Explants from Third-Trimester Human Pregnancy.

Previously, we demonstrated the expression of apelin and G-protein-coupled receptor APJ in human placenta cell lines as well as its direct action on placenta cell proliferation and endocrinology. The objective of this study was to examine the effect of apelin on placenta apoptosis in BeWo cells and villous explants from the human third trimester of pregnancy. The BeWo cells and villous explants were incubated with apelin (2 and 20 ng/mL) alone or with staurosporine for 24 to 72 h. First, we analysed the dose- and time-dependent effect of apelin on the expression of apoptotic factors on the mRNA level by real-time PCR and on the protein level using Western blot. Next, we checked caspase 3 and 7 activity by Caspase-Glo 3/7, DNA fragmentation by the Cell Death Detection ELISA kit and oxygen consumption by the MitoXpress-Xtra Oxygen Consumption assay. We found that apelin increased the expression of pro-survival and decreased proapoptotic factors on mRNA and protein levels in both BeWo cells and villous explants. Additionally, apelin inhibited caspase 3 and 7 activity and DNA fragmentation in staurosporine-induced apoptosis as also attenuated oxidative stress by increasing extracellular oxygen consumption. The antiapoptotic effect of apelin in BeWo cells was mediated by the APJ receptor and mitogen-activated protein kinase (ERK1/2/MAP3/1) and protein kinase B (AKT). The obtained results showed the antiapoptotic effect of apelin on trophoblast cells, suggesting its participation in the development of the placenta.

Diagnostic and prognostic relevance of microparticles in peripheral and uterine blood of patients with endometrial cancer.

OBJECTIVES: Exosomes - microvesicles which are secreted by living cells - can be produced from different cell types and detected in various body fluids. They are the carriers of intercellular information which regulate tumor microenvironment and are considered to be involved in tumor progression and metastasis. Cancer cells can secrete more exosomes than healthy cells, and are expected to be potential tools for tumor diagnosis and treatment. MATERIAL AND METHODS: In this report, we present the results of microparticle analysis in peripheral and uterine blood of patients with endometrial cancer. To the best of our knowledge, this study has been the first to report microvesicle status in peripheral and uterine blood samples. The aim of the study was to determine the amount of total (TF+), endothelial (CD144+) and monocytic (CD14+) microparticles. The counting of the selected microparticles in citrate plasma was performed using flow cytometry on the BD Canto II cytometer. RESULTS: We found that the total amount of microparticles in cancer patients was much higher than in healthy controls. Moreover, microparticle count in uterine blood was higher than in peripheral blood of patients with endometrial cancer. We also demonstrated that the amount of microparticles correlates with the histologic grade and clinical stage of the tumor. CONCLUSIONS: The most interesting finding in this work was the high level of TF, CD144 and CD14 MPs in uterine blood samples. Thus we can consider the monocyte-macrophage-derived MPs as a candidate marker of endometrial cancer and maybe very critical part of the endometrial carcinogenesis.

The dynamics of autonomic nervous system activity and hemodynamic changes in pregnant women.

OBJECTIVE: The purpose of this research was to assess the dynamics of autonomic nervous system(ANS) and hemodynamic activity changes during uncomplicated pregnancy. METHODS: We enrolled 36 pregnant women (mean age 29 ± 4.8 years) and a control group of 10 non-pregnant women (mean age 25.9 ± 0.88 years). The examination was performed in the 1st, 2nd, and 3rd trimester. Continuous registration of BP, ECG, and cardioimpedance was performed with Task Force Monitor 3040i. ANS activity was measured using the following parameters: HRV, BPV, BRS at rest, and in response to autonomic tests. RESULTS: Compared to the 1st trimester, an increase in HR (73 vs. 92 bpm; p < 0.001) and mean BP (80 vs. 85 mmHg, p < 0.01) was observed in the 3rd trimester. In the 1st trimester, the BRS of pregnant women was insignificantly higher than in the controls (24.8 vs. 22.3 ms/mmHg); subsequently, it decreased significantly, to 13.4 ms/mmHg in the 3rd trimester (p = 0.0004). An increase in nLF (39.57 ± 13.75 vs. 58.73 ± 15.55; p = 0.001) and LF/HF ratio (1.03 ± 0.76 vs. 1.85 ± 0.8; p < 0.00002) was revealed in HRV analysis conducted in the 3rd trimester, as compared to the 1st tri- mester, along with a decrease in nHF (60.43 ± 13.71 vs. 41.26 ± 15.55; p < 0.001). An increase in LF/HF-sBPV (1.05 ± 0.48 vs. 1.58 ± 0.44; p = 0.01) was recorded in BPV analysis at rest in the 3rd trimester as compared to the respective 1st trimester value. CONCLUSION: Our findings suggest that pregnancy is associated with dynamic changes in autonomic balance, namely doubled dominance of the sympathetic component. Hypervolemia seems the major factor responsible for autonomic and hemodynamic changes observed during pregnancy, as it causes an increase in BP and simultaneous decrease in BRS.

[The 1691 G > A (factor V Leiden) and 1328 T > C V coagulation factor polymorphisms and recurrent miscarriages]. / Polimorfizmy 1691 G > A (czynnik Leiden) i 1328 T > C genu V czynnika krzepniecia a wystepowanie poronien nawracajacych.

Objectives: Inherited thrombophilia might lead to recurrent pregnancy loss (RPL). The aim of the study was to estimate the prevalence of V coagulation factor polymorphisms related with inherited thrombophilia among women in Malopolska region.Material and methods: Group of 136 women, who experienced at least 2 unexplained, idiopathic pregnancy loss. 106 healthy women having at least one uncomplicated pregnancy and delivered healthy children constituted a control group. Each patient were examined for factor V Leiden (FVL) and mutation 1328 T>C of factor V gene with use of real ­time PCR and Taq-Man probes.Results: Among patients with RPL inhabiting region of Malopolska compared to control group occurred higher prevalence of FVL and mutation 1328 T>C. There is coincidence of polymorphism 1328 T>C of factor V gene and FVL in group of early and late RPL.Conclusions: TC genotype of 1328 T>C mutation carriers reveal tendency toward RPL below 7 weeks of pregnancy.Based on results of these findings inherited thrombophilia evaluation in patients after two or more RPL should be recommended.

Evening not morning plasma cortisol level is higher in women with polycystic ovary syndrome.

INTRODUCTION: The aim of our study was to assess the morning and evening cortisol plasma levels in women with polycystic ovary syndrome (PCOS). MATERIAL AND METHOD: 95 patients gave their informed consent to participate in the study and were divided into 2 groups. Group A consisted of 40 PCOS patients and group B consisted of 55 women without features of PCOS. Between day 5 and 8 of the menstrual cycle, morning (7 am), fasting blood samples were taken for the assessment of luteinizng hormone (LH), follicle stimulating hormone (FSH), estradiol, cortisol, prolactin, thyreotropin (TSH), testosterone and dehydroepeiandrosterone (DHEAS). Evening (5 pm) blood samples were also taken for the evaluation of plasma cortisol level RESULTS: There were no differences in mean age, body mass index (BMI), FSH, SHBG, PRL, estradiol and TSH levels between group A and group B. Mean plasma LH level was higher in group A compared to group B (10.7 ± 6.8 IU/l vs 6.6 ± 4.5 LU/l, p < 0.02). Mean plasma testosterone and DHEAS levels were also higher in PCOS patients (3.8 ± 0.6 nmol/l vs 1.63 ± 0.6 nmol/l; 427.7 ± 162.9 vs 236.6 ± 97.8 respectively, p < 0.001). Mean evening plasma cortisol level was higher in PCOS patients (11.8 ± 4.1 ug/dl vs 4.7 ± 1.3 ug/dl, p < 0.02). Mean morning plasma cortisol levels did not differ between groups. CONCLUSION: PCOS women showed the increased evening plasma cortisol level with impacted diurnal secretion rate.

The vascular architecture of the supravaginal and vaginal parts of the human uterine cervix: a study using corrosion casting and scanning electron microscopy.

The aim of this study was to visualize and describe the vascular architecture of the vaginal and supravaginal parts of the human uterine cervix. Uteri collected at autopsy (n = 42) were perfused via the afferent vessels with fixative followed by Mercox resin. After polymerization of the resin, corrosion was performed. The obtained vascular casts of the cervix, visualizing all vessels including capillaries, were examined using scanning electron microscopy. Both in the vaginal and supravaginal parts of the cervix, four distinct vascular zones were distinguished - the outer zone containing large arteries and veins, the arteriole and venule zone, the endocervical mucosal capillaries zone and the pericanalar zone containing small veins and capillaries. In the pericanalar zone ran small veins, responsible for draining the mucosal capillaries. Both in the muscular layer, as well as in the pericanalar zone, arterioles and venules passed close to each other, often adjoining. This study introduces the idea of two systems responsible for draining blood from the mucosal capillaries. It is also the first to suggest the possible existence of a countercurrent transport between adjoining veins and arteries.

[Translabial ultrasonography in pelvic floor prolapse and urinary incontinence diagnostics]. / Przezkroczowa ultrasonograficzna diagnostyka zaburzen statyki narzadu rodnego i nietrzymania moczu.

Technological advances in the construction of sonographic devices and increasingly universal access to such tests considerably widens the range of diagnostic application of the sonographic examination. This situation also refers to pelvic organs prolapse. At present, sonographic sets used in everyday obstetrical-gynecological practice allow for insight into the structures forming the female pelvic floor, and the obtained images constitute a valuable addition to the physical examination. Positioning the sonographic transducer on a the perineum enables to visualize the three compartments of the female pelvis minor. After freezing the image, it is possible to assess the position of anatomical structures in relation to bones and designated surfaces, establish mutual distances and measure appropriate angles. Most information can be obtained in this manner within the range of the frontal compartment, whose damage is often linked with urinary incontinence. The examination standards developed so far, including the analysis of the quantitative parameters, greatly minimize the potential subjectivity of the assessment of the existing disorders. Apart from its low costs, the main value of the sonographic examination of the pelvic floor is the possibility to dynamically assess the changes in statics which take place during functional testing. Not only does it have a cognitive significance, but also it allows to adjust the scope of the surgical correction to the existing damages. Thus, indirectly it can contribute to the reduction of a number of subsequent remedial surgeries. Three-dimensional sonography allows to thoroughly examine the construction and functioning of the anal levators and to detect their possible damage. It is the trauma to these muscles--occurring, among others, during childbirth--that is one of the major causes of pelvic organs descent and prolapse in women. Sonographic examination also enables to visualize the artificial material, the use of which is increasingly frequent in the surgical treatment of pelvic organs prolapse. The remedial kits currently in use are not visible in classic X-ray examinations and CT however, they can be easily visualized by means of a sonographic test. This way it is possible to locate the positioned tapes or meshes and to establish their orientation towards the urethra, the bladder and the anus. It is also possible to observe the complications (e.g. hematomas) following the surgical procedures via the transvaginal access. Transperineal sonography is a relatively inexpensive method that may be performed in almost every ultrasonographic laboratory Its value, approaching the MRI diagnostic value in the assessment of the pelvic floor defects and its low cost, place ultrasonography on the first place among the methods of imaging the pelvic and uro-genital diaphragm. The results of diagnostics of pelvic organs prolapse obtained by placing the sonographic head on the patient's perineum may also help in establishing the changes of the support apparatus for the uterus and the vagina, bladder urethra and perineum during functional testing and, thus, to verify the clinically established degree of the prolapse. It seems, also, that this method may be used during peri-urethral injections of specific substances and post-application follow-up, an increasingly frequently employed method of urinary incontinence treatment.

[Antioxidative system in pregnant women infected by Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum]. / Wplyw zakazen kanalu szyjki macicy u ciezarnych wywolanych przez Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum na uklad antyoksydacyjny.

OBJECTIVES: The study was performed to evaluate the frequency of cervical infections with Chlamydia trachomatis, Ureaplasma urealyticum and Mycoplasma hominis in women with normal pregnancy and with symptoms of spontaneous abortion or preterm delivery. Antioxidant status of pregnant women was assessed. Elevated oxidative status is claimed to be one of the factors causing threatened miscarriage and premature labor. MATERIAL AND METHODS: From 73 women endocervical culture was investigated for Chlamydia trachomatis (method of rRNA Hybridization), Ureaplasma urealyticum and Mycoplasma hominis (Mycoplasma IST 2 test). Gestational age ranged from 14-30 weeks. They were categorized into 2 groups: patients not presenting these symptoms without history of pregnancy complications and patients with threatened abortion or symptoms of preterm delivery Following oxidative status factors were measures: Superoxide Dysmutase, Catalase and Glutathion Peroxidase in blood and saliva of 49 pregnant women of both groups were elevated. Statistica: the Student's T test, chi-square test, U Manna-Whithney's test. RESULTS: The results show difficulty in assessment of direct influence of Chlamydia infection on oxidative stress. Ch. trachomatis was present in 9.6%, M. hominis in 6.8%, U. urealyticum in 28.8% of vaginal cultures. Chlamydiasis statistically increases production of Reactive Oxygen Forms what results in unfavorable outcome of the pregnancy Increase of the following parameters was observed: Superoxide Dysmutase (5583.2 vs 8980.2U/ml/sec) and Catalase (15.1 vs 18.3U/ml/sec). CONCLUSION: Infection of Chlamydia trachomatis could be a significant factor in pathogenesis of threatened abortion and preterm delivery as it increases oxidative stress.

Differences in the Composition of Gut Microbiota between Patients with Parkinson's Disease and Healthy Controls: A Cohort Study.

Gut microbiome and colonic inflammation can be associated with the predisposition and progression of Parkinson's disease (PD). The presented study aimed to compare gastrointestinal microbiota composition between patients diagnosed with PD and treated only with Levodopa to healthy controls. In this prospective study, patients were recruited in 1 academic hospital from July 2019 to July 2020. The detailed demographic data and medical history were collected using a set of questionnaires. Fecal samples were obtained from all participants. Next-Generation Sequencing was used to assess the microbiota composition. The endpoint was the difference in composition of the gut microbiota. In this study, we enrolled 27 hospitalized PD patients with well-controlled symptoms. The control group included 44 healthy subjects matched for age. Among PD patients, our results presented a higher abundance of Bacteroides phylum, class Corynebacteria among phylum Actinobacteria, class Deltaproteobacteria among phylum Proteobacteria, and genera such as Butyricimonas, Robinsoniella, and Flavonifractor. The species Akkermansia muciniphila, Eubacterium biforme, and Parabacteroides merdae were identified as more common in the gut microbiota of PD patients. In conclusion, the patients diagnosed with PD have significantly different gut microbiota profiles in comparison with healthy controls.

Metastatic and non-metastatic sentinel inguinofemoral lymph nodes in vulvar cancer show an increased lymphangiogenesis.

INTRODUCTION AND OBJECTIVE: Lymph node involvement is a strong predictor of disease recurrence and patient survival in vulvar cancer. The aim of the study was to evaluate the feasibility of sentinel lymph node (SLN) screening, the incidence of skip metastases, and lymph node lymphangiogenesis. MATERIAL AND METHODS: Fifty-five patients participated in this prospective, single centre study. A double SLN screening method was employed using radiocolloid (technetium-99 sulfur colloid) and 1.0% Isosulfan Blue. Immunohistochemistry, using a mouse monoclonal antibody against D2-40, was used to evaluate lymphatic vessel density (LVD). All calculations were performed using STATISTICA software v. 10 (StatSoft, USA, 2011); p < 0.05 was considered significant. RESULTS: Using both methods of SLN detection, 100% accuracy was achieved, and skip metastases were diagnosed in only one woman (1.82%). Peri-tumour median LVD was significantly increased compared with matched intra-tumour samples (p < 0.001), while median LVD was significantly lower in negative, compared with positive SLN, regardless of whether matched non-SLN were negative (p < 0.001) or positive (p = 0.005). Metastatic SLN exhibited significantly higher median LVD compared with matched negative non-SLN (p = 0.015), while no significant difference in median LVD was detected between positive SLN and matched positive non-SLN. However, negative SLN had a significantly higher median LVD compared with matched negative non-SLN (p = 0.012). CONCLUSIONS: SLN detection is a safe and feasible procedure in vulvar cancer. In patients without nodular involvement, SLN, compared with non-SLN, exhibited significantly higher median LVD, which may be an indication of its preparation to host metastases, and thus requires further investigation.

Significance of adding progesterone to the Risk of Ovarian Malignancy Algorithm for early stage ovarian cancer detection in patients with a pelvic mass: A single-center case-control study.

OBJECTIVE: To evaluate the clinical significance of the combination of cancer antigen-125 (CA-125), human epididymis protein 4 (HE4), and progesterone for the identification of ovarian masses in patients with suspected early stage ovarian cancer (OC). MATERIALS AND METHODS: This was a case-control, single-center study of 225 women with a pelvic mass of suspected ovarian origin, including 75 patients with Stage I/II OC and 150 controls. Diagnostic procedures included pelvic and rectal examinations, transvaginal ultrasound, evaluation of CA-125 and HE4 levels alone and in the Risk of Ovarian Malignancy Algorithm (ROMA), and a new algorithm combining ROMA and progesterone. RESULTS: Median CA-125 and HE4 levels were significantly higher in patients with OC compared with women with benign ovarian tumors, irrespective of menopausal status. The highest median progesterone levels occurred in premenopausal women with benign ovarian tumors, compared with premenopausal women with OC with or without benign ovarian disease. The combination of ROMA and progesterone was significantly more accurate at detecting OC compared with ROMA or CA-125 or HE4 alone, but only in premenopausal patients. CONCLUSION: Different algorithms should be used for diagnosing OC, and the addition of progesterone might improve the performance of ROMA for the diagnosis of pelvic masses in premenopausal women.