Pilot Study: FSHR Expression in Neuroendocrine Tumors of the Appendix.

Appendix neuroendocrine neoplasm (ANEN) treatment is based on tumor size and proliferation markers. Recently, the role of the follicle-stimulating hormone receptor (FSHR) from the clinical perspective has also been increasingly discussed. The FSHR is expressed in the endothelial cells of both intratumoral and peritumoral blood vessels, where it contributes to neoangiogenesis and blood vessel remodeling. FSHR expression is associated with a range of tumor types, such as gastrointestinal tumors, and it is not detected in healthy tissues located more than 10 mm from the tumor site or in tumor lymphatics. In this study, we evaluated the expression of FSHR and CD31 in the blood vessels of ANENs in females and males with confirmed histopathology. We conducted a quantitative analysis of the immunohistochemical reactions and found a higher number of microvessels in the mucosa and submucosa of neuroendocrine tumors in the appendix. A higher level of FSHR expression was observed in women. Future research should consider whether an elevated number of blood vessels along with a strong pattern of FSHR expression may influence future treatment strategies.

International evidence to inform decision making on implementing urgent response social protection measures.

This paper summarises evidence from a rapid review of international findings on the effects of cash transfers. The learnings were used to inform the design of urgent response social protection during the COVID-19 pandemic. The summary demonstrates that in response to widespread disruption, cash transfers have broad benefits for children, adults, and the wider economy. First, cash grants for child support have been shown to reduce hunger, increase dietary diversity, and reduce secondary school dropout. Transfers may also reduce child malnutrition. Second, there is some evidence that cash grants could encourage job search and economic activity. By contrast, there is little evidence that grants discourage adults from working, or increase spending on temptation goods (alcohol, tobacco). Third, for the wider economy, there is little evidence that grants will increase inflation, while some studies find that transfers create a fiscal multiplier and stimulate the local economy. Finally, we review evidence on design considerations and find that unconditional cash transfers (UCTs) are particularly well suited to rapid response when compared to conditional cash transfers (CCTs). Outside crisis settings, there is some evidence that getting recipients to enrol children in school or attend health check-ups improves these outcomes more than unconditional grants. However, the differences are small and the additional costs of implementing conditions outweigh these benefits in circumstances where response is required urgently. Comparing evidence on cash grants and food vouchers suggests both achieve similar improvements in nutrition; however, cash transfers are likely to be more cost-effective for governments, especially where a system to distribute grants is already set up.

Estrogen and Progesterone Receptor Immunoexpression in Fallopian Tubes among Postmenopausal Women Based on Time since the Last Menstrual Period.

Existing data on the expression of estrogen receptor (ERα) and progesterone receptor (PR) in fallopian tubes in postmenopausal women are mostly inconclusive. Therefore, we assessed ERα and PR immunoexpression in the oviducts of these women. One hundred postmenopausal women were divided into three groups based on time elapsed since the last menstrual period: (A) 1-5 years, (B) 6-10 years, and (C) ≥11 years. In all groups, both in the glandular epithelium and stroma of the ampulla and isthmus of the oviduct, immunolocalization of ERα and PR were noted. The glandular epithelium of the ampulla showed a higher percentage of PR-positive cells than the isthmus in each group. Regarding ERα, there were no significant differences. In the glandular epithelium in both the ampulla and isthmus, the percentage of ERα- and PR-positive cells was significantly higher than that in the stroma in each study group and higher in the A group than in the C group. In conclusion, in postmenopausal women, time elapsed since the last menstrual period in the fallopian tubes was positively correlated with the following: (1) the epithelium showed vacuolation of cytoplasm with greater frequency, (2) the proportion of ciliated cells decreased, and (3) the percentage of ERα- and PR-positive cells also decreased. The obtained results indicate a significant decrease in ERα and PR expression depending on the time that has elapsed since the last menstruation, which is undoubtedly related to the loss of the reproductive function of the patients.

Markers of Inflammation and Vascular Parameters in Selective Progesterone Receptor Modulator (Ulipristal Acetate)-Treated Uterine Fibroids.

The exact mechanism of selective progesterone receptor modulator action in leiomyoma still challenges researchers. The aim of the study was to assess the effects of ulipristal acetate (UPA) on immunoexpression of inflammatory markers and vascularization in fibroids. UPA-treated patients were divided into three groups: (1) good response (≥25% reduction in volume of fibroid), (2) weak response (insignificant volume reduction), (3) and no response to treatment (no decrease or increase in fibroid volume). The percentage of TGFß, IL6, IL10, CD117, and CD68-positive cells were significantly lower in the group with a good response to treatment vs. the control group. Moreover, the percentage of IL10 and CD68-positive cells in the group with a good response to treatment were also significantly lower compared to the no response group. Additionally, a significant decrease in the percentage of IL10-positive cells was found in the good response group vs. the weak response group. There were no statistical differences in the percentage of TNFα-positive cells and vessel parameters between all compared groups. The results of the study indicate that a good response to UPA treatment may be associated with a decrease of inflammatory markers, but it does not influence myoma vascularization.

Initial assessment of suitability of MCF-7 and HepG2 cancer cell lines for AQP3 research in cancer biology.

Cancer cell lines are widely used as in vitro models to elucidate biological processes in cancer, and as a tool to evaluate anticancer agents. In fact, the use of an appropriate cancer cell line in cancer research is crucial for investigating new, potential factors involved in carcinogenesis. One of them is aquaporin-3 (AQP3), which is a small, hydrophobic, integral membrane protein with a predominant role in water and glycerol transport. Recently, altered expression of AQP3 has been reported in many types of cancer. Increasing evidence strongly suggests that AQP3 plays a key role in cancer cell proliferation, migration and invasion. In this study, we performed an insightful characteristic of AQP3 location and its protein expression in MCF-7 human breast adenocarcinoma and HepG2 hepatocellular carcinoma cell lines in the context of cancer biology using immunocytochemistry, immunofluorescence and Western blot analyses. AQP3 was found to be located in the cell membrane and cytoplasm of MCF-7 cells, and in the cytoplasm and nuclear membrane of HepG2 cells. Immunoblotting of proteins derived from both cell lines revealed a clear band with a molecular weight of approx. 30 kDa representing an unglycosylated form of AQP3. However, the expression of this protein was higher in MCF-7 than in HepG2. Concluding, our results clearly indicated variability in both the expression levels and subcellular location of the AQP3 protein in MCF-7 and HepG2 cell lines. This leads to the possibility that the expression patterns and subcellular location of AQP3 in the tested cancer cell lines are tissue-of-origin specific, and may be related to the aggressiveness of cancer cells and their mobility.

Markers of Cellular Proliferation, Apoptosis, Estrogen/Progesterone Receptor Expression and Fibrosis in Selective Progesterone Receptor Modulator (Ulipristal Acetate)-Treated Uterine Fibroids.

There appear to be very few data on the exact mechanisms of a selective progesterone receptor modulator action in myomas. The aim of the study was to assess the effects of ulipristal acetate (UPA) on fibroids, especially on estrogen receptor (ER) and progesterone receptor (PR) immunoexpression, proliferation, apoptosis and tissue fibrosis, and to compare the above parameters in untreated (surgical attention only) and UPA-treated leiomyomas. UPA-treated patients were divided into three groups: (1) good response (≥25% reduction in volume of fibroid), (2) weak response (insignificant volume reduction) and (3) no response to treatment (no decrease or increase in fibroid volume). The study observed a significant decrease in the percentage of collagen volume fraction and ER and PR immunoexpression in the good response group, in the percentage of proliferating cell nuclear antigen (PCNA)- and Ki67-positive cells in the groups with good and weak reactions vs. control group; significantly higher apoptotic index (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive cells) in the good reaction group vs. control group. The results of the study indicate that a good response to UPA, manifested by a volume reduction of myoma, may be associated with a decrease in fibrosis, ER/PR and PCNA and Ki67 immunoexpression and an increase in cell apoptosis within the myoma.

The Effects of Long-Term Immunosuppressive Therapies on the Structure of the Rat Prostate.

Background: Little is known about the overall impact of immunosuppressive drugs on the prostate. The study aimed to determine the impact of different protocols of immunosuppressive treatment on the structure of the rat ventral prostate. Methods: For 6 months, 48 male Wistar rats received immunosuppressive drugs: cyclosporin A, tacrolimus, mycophenolate mofetil, rapamycin, and prednisone, according to three-drug protocols. Light and transmission electron microscopic studies, and quantitative evaluation of immunohistochemical expression of selected intermediate filaments, CD117+ mast cells, and CD138+ plasma cells were performed in the rat ventral prostate. Results: In all experimental groups, acini focal hyperplasia, changes to the ultrastructure of the glandular epithelium, changes in the expression of cytokeratins and desmin, and numerous mast and plasma cells in the prostate stroma were found. In cyclosporine-A-based groups, atrophy and numerous intracellular vacuoles were observed. In groups where a three-drug treatment was replaced with rapamycin, morphological alterations were less severe compared to those without conversion. Conclusions: In the rat ventral prostate, (1) immunosuppressive protocols affect the morphology and immunohistochemical expression of intermediate filaments, (2) morphological alterations, expression, and localization of selected proteins are not connected with adenocarcinoma development, and (3) conversion of the treatment to rapamycin may prevent hyperplastic abnormalities.

Investigating cellular location of aquaporins in the bovine kidney. A new view on renal physiology in cattle.

To date, 13 aquaporin isoforms (AQPs) have been discovered in mammals, of which as many as 9 are located in epithelial cells lining the individual sections of the nephron and collecting tubules. Detailed analysis of the location and expression of AQPs in the kidneys of laboratory animals and humans allowed to define the key role of these proteins in renal excretion of water and other small molecules. Unfortunately, despite the significant advances in knowledge in this area, still little is known about this subject in livestock, including cattle. Therefore, the aim of the study was to determine the expression and AQPs location in the nephron segment in the bovine kidney by immunohistochemistry and Western blot. The distribution of a total of 8 aquaporins was determined as a result of the conducted experiments. The results obtained in the present study clearly indicate that aquaporins in cattle are involved in the renal regulation of water excretion and maintenance of proper acid-base balance. Undoubtedly, changes in the distribution and expression of AQPs in bovine kidneys may be the cause of water balance disorders and disruption of the normal body fluid composition. Kidney diseases in cattle are poorly described in veterinary medicine. Knowledge of cellular location and expression of all AQPs in the bovine kidney under normal physiological condition allows a deeper understanding of the renal regulation of body homeostasis. It creates new perspective for diagnosis and pharmacotherapy in cattle in the future.

Effects of an immunosuppressive treatment on the rat prostate.

The aim of this study was to determine the influence of different combinations of immunosuppressive drugs on the morphology, ultrastructure, and expression of proliferating cell nuclear antigen and cytoskeleton proteins in the rat dorsolateral prostate. The studies were conducted on 48 male Wistar rats. The animals were divided into eight groups: a control group and seven experimental groups. For 6 months, the animals in the experimental groups were administered a combination of drugs including rapamycin (Rapa), cyclosporin A, tacrolimus (Tac), mycophenolate mofetil, and prednisone (Pred), according to the standard three-drug regimens for immunosuppressive therapy used in clinical practice. An evaluation of the morphology and ultrastructure was conducted, and a quantitative evaluation of the expression of proliferating cell nuclear antigen and desmin- and cytokeratin-positive cells with weak, moderate, and strong expression was performed. The combination of Rapa, Tac, and Pred caused the smallest morphological and ultrastructural changes in the rat prostate cells. In the case of rats whose treatment was switched to Rapa monotherapy, a decreased percentage of proliferating cells of both the glandular epithelium and the stroma was found. Decreases in body weight and changes in the expression of cytokeratin and desmin were observed in all the experimental rats. The combination of Rapa, Tac, and Pred would seem to be the most beneficial for patients who do not suffer from prostate diseases. Our results justify the use of inhibitors of the mammalian target of Rapa in the treatment of patients with prostate cancer. The changes in the expression of cytoskeleton proteins may be the result of direct adverse effects of the immunosuppressive drugs, which are studied in this article, on the structure and organization of intermediate filament proteins.

Analysis of the Relationship between Estradiol and Follicle-Stimulating Hormone Concentrations and Polymorphisms of Apolipoprotein E and LeptinGenes in Women Post-Menopause.

BACKGROUND: Menopause is the permanent cessation of menstruation due to loss of ovarian follicular activity. A review of the available literature indicates that correlations between the changes that take place in a woman's body after menopause and different genetic variants are still being sought. METHODS: The study was conducted in 252 women who had completed physiological menopause. The women were divided into groups according to the time elapsed since menopause. The total concentrations of estradiol and follicle-stimulating hormone were determined by means of electrochemiluminescence. The apolipoprotein E (APOE) and lepitn (LEP) genotypes were determined by real-time PCR and polymerase chain reaction-restriction fragment length polymorphism, respectively. RESULTS: We observed that people with the APOE3/E3 genotype entered menopause insignificantly later compared to other genotypes. Additionally, in the group of patients with the APOE3/E3 genotypes, differences in the E2 concentration were significantly related to the time since their last menstruation. There is no association found in the literature between these polymorphisms of the LEP gene and hormones. CONCLUSIONS: To date, attempts to formulate a model describing the association between E2 and FSH concentration with the polymorphisms of various genes of menopause in women have not been successful. This relationship is difficult to study because of the number of nongenetic factors. Environmental factors can explain variation in postmenopausal changes in hormone levels.

Hormone concentration, metabolic disorders and immunoexpression of androgen and estrogen-alpha receptors in men with benign prostatic hyperplasia and testosterone deficiency syndrome.

INTRODUCTION: A slight decrease in blood testosterone level in men is a physiological state associated with the aging. The aim of our study was to evaluate the occurrence of hormone and metabolic disorders, as well as the immunolocalization and immunoexpression of androgen receptors (AR) and estrogen-alpha receptors (ERa) in the prostates of men with benign prostatic hyperplasia (BPH) and coexisting testosterone deficiency syndrome (TDS). MATERIAL AND METHODS: The study involved 150 men, diagnosed with and receiving pharmacological treatment for BPH. Concentrations of glucose, total cholesterol (TCh), high-density lipoproteins (HDL), low-density lipoproteins (LDL), and triglycerides (TG) were determined in blood serum. Serum concentrations of total testosterone (TT), free testosterone (FT), estradiol (E2), luteinizing hormone (LH), insulin (I), sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS), and insulin-like growth factor 1 (IGF-1) were measured by ELISA. The number of AR-positive cells and ERa-positive cells were measured in prostate sections of men with BPH. RESULTS: Patients eligible for transurethral resection of the prostate and TDS were significantly more likely to have higher abdominal circumference and higher serum levels of insulin and IGF-1 as well as lower levels of FT and SHBG than control subjects with BPH and no TDS. Quantitative analysis revealed 35.8% AR-positive colum-nar epithelial cells and 24.3% AR-positive stromal cells in prostates of BPH patients with TDS and 30.5% and 23.0%, respectively, in BPH patients without TDS. However, the differences between the study and the control groups were statistically not significant. In prostates of BPH patients with TDS the immunoexpression of ERa was observed in 2.88% of the columnar epithelial cells and 0.39% of stromal cells. In BPH patients without TDS ERa-positive cells were only found in 0.04% of columnar epithelial cells and 0.62% of prostatic stromal cells. CONCLUSIONS: Considering the statistically significantly higher levels of I and IGF-1 and larger abdominal circumference of men with BPH and TT deficiency, it can be supposed that visceral obesity and carbohydrate disorders may contribute to the reduction of testosterone concentration. The results of our study indicate a relationship between TT concentration in the plasma of patients with BPH and the percentage of AR-positive cells in the prostate.

Caloric restriction increases ratio of estrogen to androgen receptors expression in murine ovaries--potential therapeutic implications.

Both estrogens and androgens are involved in the development and normal functioning of the ovaries. It is also known that ovarian function is regulated by diet. The goal of this study was to estimate the expression of sex hormone receptors in ovaries of mice that were on a 9-month caloric restriction (alternate-day feeding) as compared to normal control animals fed ad libitum. We found that prolonged caloric restriction in mouse ovaries led to increased expression of estrogen receptors (ERs) but did not affect expression of the androgen receptor (AR). This increase in ER:AR ration as result of caloric restriction may lead to higher sensitivity to estrogens and upon return to normal diet may increase ovulation. Thus our observation shed more light on a role of beneficial effect of calorie restriction on female reproduction.

Can metabolic disorders in aging men contribute to prostatic hyperplasia eligible for transurethral resection of the prostate (TURP)?

PURPOSE: The aim of this study was to evaluate the incidence and severity of metabolic disorders occurring in the metabolic syndrome in patients with benign prostatic hyperplasia eligible for surgical treatment. METHODS: The study group consisted men with diagnosed benign prostatic hyperplasia. The control group consisted patients recruited from basic health care units. Abdominal circumference, body weight and blood serum metabolic parameters were determined in the experimental and control groups. The concentrations of glucose were determined, as well as total cholesterol (ChT), low-density lipoprotein (LDL), high density lipoprotein (HDL) and triglycerides (TAG), by spectrophotometric method using reagent kits. RESULTS: In the study group 91 (60.3%) cases of metabolic syndrome (MetS) were diagnosed, while in the control group 71 (46.1%) men met the diagnostic criteria for this syndrome (p=0.018). The analysis shows a relationship between MetS in patients with BPH and concentration glucose, ChT, LDL, HDL, systolic blood pressure and diastolic blood pressure. We found no significant statistical relationship between body weight, abdominal circumference and concentration TAG, hypertension in patients and controls. CONCLUSIONS: in the study presented in this article, statistically significant relationships between BPH and the diagnostic parameters of the metabolic syndrome were demonstrated. These results indicate to the necessity of the modification of the lifestyle, taking preventive measures in diabetes, and evaluation of lipid metabolism disorders. It is recommended to assess symptoms that may suggest BPH (as a manifestation of LUTS) in men over 50 years of age with diagnoses of metabolic disorders (including MetS), and provide them with specialist urological care in order to prevent surgical treatment of the prostate.

The influence of immunosuppressants on the morphology, proliferating cell nuclear antigen (PCNA) and apoptosis in the rat ventral prostate.

AIM: Analysis of the impact of immunosuppressants on apoptosis and PCNA in the rat ventral prostate. METHOD: The studies were performed on 48 male Wistar rats. The animals were divided into a control group and 7 experimental groups. For 6 months, the rats were administered drugs such as: rapamycin (Rapa), cyclosporin A (CsA), tacrolimus (Tac), mycophenolate mofetil (MMF) and glucocorticosteroids (GS). During section of the rats, prostate ventral lobes were obtained. Morphological evaluation (HE, PAS), TUNEL assay, PCNA expression analysis and quantitative image computer analysis were performed. RESULTS: The highest percentage of apoptosis in epithelial cells was observed in groups which received two combinations of drugs: (V) CsA, MMF, GS and (VII) Tac, MMF, GS. A much lower percentage of apoptotic epithelial cells was found in the groups where the treatment schemes included rapamycin throughout the duration of the study. Interestingly, the conversion of the treatment to rapamycin caused a significant reduction of apoptosis in epithelial cells as well as in proliferation in both epithelial and stromal cells. CONCLUSIONS: On one hand, the obtained results may explain the anticancer activity of rapamycin in reducing the proliferation of epithelial cells, and on the other hand the adverse effect of rapamycin in the form of reduced regeneration of these cells. Taking into account the prostate in isolation from other organs/systems, the dosage scheme with Rapa, Tac and GS would appear to be the most favorable, due to the smallest morphological changes.