Ekspresja izoform VEGF i ich receptorów w mięśniakach macicy kobiet

Objectives: The aim of the study was to determine the expression of VEGF (vascular endothelial growth factor) isoforms and their receptors in uterine myomas. Material and methods: The study included 40 women with myomas of reproductive age and 40 perimenopausal women (the study group). Myometrial samples (the control group) were taken from 10 women undergoing hysterectomy for ovarian tumors and 10 older women undergoing hysterectomy for uterine prolapse. Results: A significantly increased expression of VEGF-A has been found in myomas, both small and large, in the younger women, which may by a sign of increased angiogenesis and intensive tumor growth. In perimenopausal women, the increase of VEGF expression was observed only in the endothelium and vascular smooth muscle. Conclusion: An important conclusion of this study is that angiogenesis is independent of myoma size, which may suggest intensive tumor growth and the related increased angiogenesis. High expression of VEGF-A and VEGF-R1 receptors in large myomas can probably cause malignant transformation and more extensive growth, regardless of patient age.Cel pracy: Celem pracy było próba określenia ekspresji izoform VEGF (ang. vascular endothelial growth factor) i ich receptorów w mięśniakach macicy. Materiał i Metody: Badania przeprowadzono na 40 chorych z mięśniakami w wieku rozrodczym i 40 chorych z mięśniakami w okresie okołomenopauzalnym. Grupę kontrolną stanowiły próby pobrane od 10 kobiet poddanych histerektomii z powodu guzów jajnika i 10 starszych kobiet poddanych histerektomii po wypadnięciu macicy. Wyniki: Stwierdzono statystycznie wyższą ekspresję VEGF-A w komórkach mięśniaków małych i dużych u kobiet młodych. Może to skazywać na zwiększoną angiogenezę, a co za tym idzie wzmożony wzrost guza. U kobiet w wieku okołomenopauzalnym wzrost ekspresji VEGF stwierdzono jedynie w śródbłonku i mięśniówce naczyń. Wniosek: Ważnym podsumowaniem tej pracy, jest stwierdzenie, że angiogeneza jest niezależna od wielkości mięśniaka. Można zatem wnioskować, że wzmożony wzrost guza związany jest ze zwiększoną angiogenezą. Wysoka ekspresja VEGF-A i receptora VEGF-R1 w mięśniakach dużych, może być powiązana ze wzrostem złośliwości komórek i ich intensywnym rozwojem niezależnie od wieku kobiety.

Immunolocalization of ABC drug transporters in human placenta from normal and gestational diabetic pregnancies

Objectives: ABC transporters, P-gp, MDR3, BCRP and MRP1, can bind both endo- and exogenous ligands. The latter include immunosuppressive, anticancer, sedative, anticonvulsant, antiparasitic and cardiovascular drugs, as well as HIV protease inhibitors and antibiotics. Protein transporters are also involved in tissue distribution of orally administered medicines in combination therapy for gestational diabetes mellitus (GDM) and could be used during GDM treatment. The distribution depends on transporter specificity, its expression and subcellular localization. The aim: The aim of the study was to compare P-gp, MDR3, BCRP and MRP1 localization in placental tissues from normal and GDM diabetic pregnancies. Material and methods: Tissue samples were taken from 10 normal and 10 GDM placentas. Immunohistochemical reactions were performed with the use of adequate monoclonal antibodies. Avidin-biotin-peroxidase complex method was used for the visualization of antigen-antibody complexes. Results: P-gp, MDR3 and BCRP were found in all parts of normal human placenta i.e. the amniotic epithelium, cytotrophoblast, syncytiotrophoblast and decidual cells. P-gp and BCRP, but not MDR3 and MRP1, were also localized on the endothelial cells of fetal blood vessels in the chorionic plate, as well as stem and tertiary villi. MRP1 expression was observed in the cytotrophoblast and the syncytiotrophoblast. Its expression was very low or undetectable in the amniotic epithelium and the majority of decidual cells. Immunohistochemical reactions within the syncytiotrophoblast showed apical (P-gp, BCRP), apical and basal (MRP1) or diffuse (MDR3) distribution. The main changes observed in GDM placentas included weaker MRP1 and MDR3 positive reactions in the syncytiotrophoblast, slightly lower expression of P-gp in the decidual and amniotic epithelial cells, and MDR3 in the amniotic epithelium. Conclusions: Our results indicate that GDM-related changes in the environment of placental cells do not substantially influence tissue and subcellular location of ABC transporters. Nevertheless, the expression of P-gp, MDR3 and MRP1 may be lower in comparison to normal placentas. Basal syncytiotrophoblast transporters, MRP1 and MDR3, seem to be more sensitive to the influence of GDM than apical proteins, what may result in altered biodisposition of endogenous substrates and drugs

The aromatase expression in myomas and myometriums of women in reproduction and perimenopausal age.

Uterine myomas represent one of the most common female pathologies. Uterine smooth muscle myomas or fibromas are benign tumours which respond to hormones and their etiology induces wide interest. The myomas were found to contain aromatase and, in addition, cells of the myomas were found to synthesize estrogen. This study was conducted on patients with the myomas, in either generative age or in the perimenopausal period. Expression of aromatase was detected in patients of various age, with large or small uterine myomas, using an immunohistochemical technique. In addition expression of the enzyme was examined at the periphery of every myoma

Immunohistochemical localization of selected pro-inflammatory factors in uterine myomas and myometrium in women of various ages

Uterine myomas represent one of the most frequently manifested benign tumors in women. They originate from smooth muscle cells of myometrium or its blood vessels. Many studies suggest that inflammation and pro-inflammatory factors may play a role in the carcinogenesis with an involvement of the transcription factor NF-kappaB which activity can be controlled by various environmental factors, including many cytokines. The aim of the study was to investigate the expression of NF-B, interleukin-1β (IL-1β), tumor necrosis factor a (TNF-α), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) in myometrium and uterine myomas of women of various age. The expression of NF-kappaB, selected cytokines and enzymes was estimated in women of reproductive or perimenopausal age by semiquantitative immunohistochemistry. The expression of the examined proteins was higher in myomas than in control myometrium and was dependent on the size of myomas and the age of women. However, the expression of the cytoplasmic NF-kappaB observed in uterine myomas was independent on the size of myomas and no significant differences were observed in the number of stained nuclei between control and myoma groups. Thus, the expression of proinflammatory factors in myomas was not accompanied by the nuclear activation of NF-kappaB p65. The results of our study indicate that the examined factors may be involved in the pathogenesis of benign tumors and not only malignant diseases. (Folia Histochemica et Cytobiologica 2013, Vol. 51, No. 1, 73–83

The relation between plasma α-synuclein level and clinical symptoms or signs of Parkinson's disease

Introduction Parkinson disease (PD) is the common neurodegenerative disease. α-Synuclein (ASN), main aggregating protein in neural cells of CNS in PD, was found in peripheral fluids. Testing ASN in plasma is potential test for diagnose PD, but previous studies are controversial. The aim of this study was to investigate if plasma ASN level may be a valuable biomarker, is the level of plasma ASN concentration different in various motor subtypes of diseases, is there a relation between the level of plasma ASN and the severity of motor symptoms. Methods Patients with PD hospitalized in Neurology Department, Medical College were performed sequencing the 8th and 9th exon of GBA gene. Next plasma ASN level was tested in 58 patients with sequenced GBA gene and in 38 healthy volunteers (HV), matched by the age (respectively 68.43 vs. 64.57 years of age) and sex (female %, respectively: 43.10 vs.44.74). Patients were assessed with the scales: UPDRS (II, III, IV), Hoehn–Yahr (HY) and qualified to PIGD or TD subtype. For homogeneity of the group patients with GBA mutation were excluded from the analysis. Results The ASN level did not differ between patients and HV (respectively: 4.53 vs. 3.73ng/ml) and between patients with different subtypes. There was inverse correlation between ASN and HY in PIGD subtype. Conclusions Plasma ASN level is not valuable marker of the disease. It does not differ in subtypes of the disease. There is relation between plasma ASN level and the severity of the disease in PIGD subtype

The TRAF2 and TRAF6 expression in myomas and myometrium of women in reproduction and perimenopausal age.

Uterine myomas represent one of the most common female diseases. Uterine myomas or fibromas are benign, hormone-responding tumours of, respectively, smooth muscles and fibroblasts and their aetiology induces a significant interest. In myomas the presence of aromatase was detected and, in addition, oestrogen was found to be synthesized in myoma cells. The studies were performed on myoma patients of generative age and those in peri-menopausal age. Expression of TRAF2 and TRAF6 proteins was examined using immunohistochemistry and Western blot approach in small and large uterine myomas isolated from women of various age. In addition, the evaluation was conducted at the periphery of every myoma. We indicated that the level of both tested proteins in myomas is higher than in control. TRAF2 level in myometrium was lower than in myomas but higher than in control. In the case of TRAF6 those changes were ambiguous. Age didn't have influence the level of expression in both tested TRAF in studied structures

Immunohistochemical visualization of pro-inflammatory cytokines and enzymes in ovarian tumors

Epithelial ovarian cancer represents one of the most deadly gynaecological neoplasms in developed countries and is a highly heterogeneous disease. Epidemiological studies show that anti-inflammatory drugs reduce the incidence and mortality of several types of cancer, indicating the potential role of pro-inflammatory factors in carcinogenesis. The expression of pro-inflammatory factors in various cancer types, including ovarian cancer, was assessed in many studies, yielding in consistent results, often due to the histological heterogeneity of various cancers. The aim of the study was to investigate the expression of IL-1, IL-6, TGF-β, TNF-α, COX-2,iNOS, and NF-kB in serous and mucinous ovarian cancers. Ninety cases of ovarian tumors classified into mucous and serous type (45 patients in each group) were selected. Each group was classified into subgroups according to the three stages of tumor differentiation, i.e. into (i) benign, (ii) borderline and (iii) malignant tumors. The presence of proteins of interest in paraffin sections was analysed by immunohistochemistry. The expression of most of the studied factors depended on the histological tumor subtype and the degree of malignancy. Expression of NF-κB appears to be related to the level of the neoplastic differentiation only in the group of serous tumors, while the presence of IL-6 in the mucinous tumor subtype was observed only in the case of benign lesions. Expression of IL-1, TNF-α and COX-2 increased with the stage of the disease in both serous and mucinous tumors. The highest level of TGF-β expression was observed in serous borderline tumors. The different levels of iNOS immunoreactivity between the groups of serous and mucinous tumors were observed only in borderline tumors. The results of our study may be helpful in designing therapeutic strategies depending on the type of ovarian cancer

Changes in the subcellular and tissue location of estrogen and progesterone receptors in rat uterus after long-term treatment with analogs of gonadoliberin

Objectives: Certain therapies with the use of analogs of gonadotropin-releasing hormone (GnRH, gonadoliberin) aim at achieving the effect of desensitization of the pituitary gland that causes inhibition of the hypothalamicpituitary-gonadal axis. The resulting hormonal changes may influence the location and expression of estrogen and progesterone receptors, as well as their endogenous functions. The aim: The aim of the study was to investigate whether long-term administration of low doses of dalarelin (GnRH agonist) and cetrorelix (GnRH antagonist) affected subcellular and tissue-specific location of ERα and ERβ estrogen receptors and progesterone receptor (PR) in rat uterus, as well as explore the extent to which the changes were reversible. Material and methods: Analogs were administered to SPD adult females in the course of 3 months, at a dose of 6 μg/kg b.w. Afterwards, the ovaries and the uterus were resected – in the course of 4 weeks after treatment completion. Tissue paraffin-embedded samples were stained with hematoxyline-eosin for morphological studies or incubated with specific antibodies for the immunohistochemical studies (ABC method). Results: GnRH analogs induced desensitization, resulting in specific and relatively persistent histological changes in the ovaries and the uterus. Strong nuclear reaction for ERα in the lining and the glandular epithelial cells in dalarelintreated rats, and lack of expression changes in cetrorelix-treated rats, were observed in the uterus. Epithelial ERα expressions were accompanied by diminished ERβ and elevated PR expression, as well as diminished ERα andERβ expression, and unchanged PR expression in the stromal and muscle cells, in both dalarelin- and cetrorelixtreatedrats. The majority of the changes were reversible after treatment discontinuation. Conclusions: Long-term exposure to low doses of GnRH analogs causes morphological changes in the uterine tissues, accompanied by reversible changes of the ERα, ERβ and PR expression, possibly influencing tissue sensitivity. These changes indicate that agonist and antagonist regulate ERα expression by means of different mechanisms. A functional interaction between the receptors, depending on ERβ expression, direct influence of analogs on the local hormonal axes, and dose-dependent effects, cannot be excluded. After discontinuation of the analog treatment, the time needed for stabilization of ER and PR expression is shorter than the period of time required to restore histological structure of the uterus