Immunohistochemical detection of follicle stimulating hormone receptor (FSHR) in neuroendocrine tumours

Wstęp: Wiadomo, że ekspresja receptorów folitropiny (FSHR) występuje w gonadach i wywodzących się z gonad nowotworach. Ostatniowykazano ich obecność także w innych nowotworach endokrynnych, takich jak guzy nadnerczy i gruczolaki przysadki. Ponadtostwierdzono immunopozytywność dla FSHR w śródbłonkach około- i wewnątrzguzowych naczyń krwionośnych różnych nowotworówzłośliwych. W obecnej pracy wykazano obecność FSHR zarówno w komórkach nowotworowych, jak i niektórych wewnątrz-guzowychnaczyniach krwionośnych guzów neuroendokrynnych (NETs).Materiał i metody: Zbadano 16 wycinków pobranych od 14 pacjentów z NETs. Odczyny immunohistochemiczne wykonano z użyciemprzeciwciała skierowanego przeciw fragmentowi 1-190 ludzkiego FSHR oraz przeciwciała dla Ki-67.Wyniki: Dodatni odczyn z przeciwciałem anty-FSHR stwierdzono w cytoplazmie większości komórek badanych guzów. W połowie badanychNETs odnotowano także immunopozytywność dla FSHR w śródbłonkach wewnątrzguzowych naczyń krwionośnych. Dodatniodczyn dla FSHR obserwowano częściej w naczyniach krwionośnych nowotworów z wyższym indeksem Ki-67.Wnioski: Ektopowe FSHR, jeśli są one aktywne, mogą przekazywać sygnały nasilające dalszy wzrost NETs.(Endokrynol Pol 2013; 64 (4): 268–271)Introduction: Follicle stimulating hormone receptors (FSHR) are well known to be expressed in gonads and in gonadal tumours. Recently,their incidence has also been revealed in endocrine non-gonadal tumours such as adrenal and pituitary tumours. Moreover, FSHR immunostaininghas also been reported in endothelium of intra- and peritumoral blood vessels of a large series of cancers. The presentpaper reports on the incidence of FSHR in both tumoral cells and some intratumoral blood vessels of neuroendocrine tumours (NETs).Material and methods: Sixteen NETs samples were taken from 14 patients. The tumour samples were immunostained using the antibodyraised against 1-190 amino acid sequence from the human FSH-R and anti-Ki67 antibody.Results: In all the samples examined, the majority of tumoral cells were immunostained with anti-FSHR antibody. Positive immunostainingconcerned also the intratumoral blood vessels endothelia in a half of the examined samples. Immunopositive blood vessels were foundmore often in tumours with higher Ki-67 index.Conclusion: FSHR expressed in NETs, if they are functional, may mediate the signals which can enhance further tumour growth

Wpływ zahamowania syntazy tlenku azotu na wywołane dietylstilbestrolem hiperprolaktynemię i rozwój guza przysadki

Introduction: The overexpression of nitric oxide synthase (NOS) has been found in tumours, including pituitary adenomas. It has also been found that NOS is overexpressed in human spontaneous pituitary adenomas. The question arises whether NOS and its product, nitric oxide (NO), are involved in pituitary tumourigenesis. To investigate this question, in the present paper we examine the effects of NOS inhibition on the development of diethylstilbestrol (DES)-induced prolactin–secreting pituitary tumours in rats. Material and methods: Thirty male Fisher 344 rats, four weeks old, were submitted to subcutaneous implantation of a silastic capsule containing DES (10 mg/capsule) or of an empty capsule. Six weeks after implantation, some of the DES-treated animals received a NOS inhibitor, N-nitro-l-arginine methyl ester (NAME), 1 mg/mL, in their drinking water, for the subsequent 14 days. Eight weeks after the implantation, all the animals were sacrificed, their pituitaries were weighed, and samples of heart blood were collected for prolactin (PRL) and vascular endothelial growth factor (VEGF) measurements. Results: It was found that DES implantation significantly increased pituitary mass, as well as PRL and VEGF concentrations in blood serum. On the other hand, the administration of NAME did not affect significantly either VEGF concentration or pituitary mass. On the other hand, it did induce a further increase in PRL levels. Conclusions: These findings indicate that NO is involved in oestrogen-induced hyperprolactinaemia, but does not play a crucial role in oestrogen-induced pituitary tumourigenesis. (Pol J Endocrinol 2012; 63 (2): 115–118)Wstęp: W licznych nowotworach, w tym także gruczolakach przysadki, stwierdzono zwiększoną ekspresję syntazy tlenku azotu (NOS). Powstaje pytanie, czy NOS oraz jej produkt — tlenek azotu (NO) biorą udział w patogenezie guzów przysadki. W celu zbadania tego zagadnienia autorzy niniejszej pracy prześledzili wpływ zahamowania aktywności NOS na rozwój prolaktynowego guza przysadki wywołanego dietylstilbestrolem (DES) u szczurów. Materiał i metody: Czterotygodniowym szczurom samcom szczepu Fisher 344 implantowano podskórnie silastikowe kapsułki zawierające 10 mg DES w kapsułce lub puste kapsułki. Sześć tygodni po implantacji część zwierząt, którym implantowano kapsułki z DES, otrzymywała inhibitor NOS — ester metylowy N-nitro-l-argininy (NAME) w wodzie do picia, w stężeniu 1 mg/ml, przez 14 dni. Osiem tygodni po implantacji kapsułek zwierzęta uśmiercono, pobrano i zważono przysadki oraz próbki krwi z serca w celu oznaczenia stężeń prolaktyny (PRL) i czynnika wzrostu śródbłonków naczyniowych (VEGF). Wyniki: Stwierdzono, że implantacja DES znamiennie zwiększa masę przysadki oraz stężenia PRL i VEGF w surowicy krwi. Równoczesne podawanie NAME nie ma istotnego wpływu na masę przysadki ani stężenie VEGF, natomiast powoduje dalszy wzrost stężenia PRL. Wnioski: Uzyskane wyniki wskazują, że NO wpływa na hiperprolaktynemię indukowaną DES, ale nie odgrywa istotnej roli w rozwoju guza prolaktynowego indukowanego estrogenem. (Endokrynol Pol 2012; 63 (2): 115–118

Three administrative texts from the time of the Third Dynasty of Ur in an anonymous collection in Poland

The article is a full edition (photography, autography, transliteration, translation and commentary) of three previously unpublished Neo-Sumerian administrative documents, which are in one of the anonymous collections in Poland. The tablets come from the two provincial archives of the kingdom of Third Dynasty of Ur - Puzriš-Dagan and Girsu-Lagaš, and their content is typical of this group of cuneiform texts.The article is a full edition (photography, autography, transliteration, translation and commentary) of three previously unpublished Neo-Sumerian administrative documents, held in one of the anonymous collections in Poland. The tablets come from two provincial archives of the kingdom of the Third Dynasty of Ur, Puzriš-Dagan and Girsu-Lagaš, and their content is typical of this group of cuneiform texts

Immunohistochemical detection of FSH receptors in pituitary adenomas and adrenal tumors

<p><strong> </strong></p><p><strong><em>Objectives</em></strong>. Follicle stimulating hormone (FSH) receptors (FSHR) are physiologically expressed in the ovary and testis. It is well known that FSHR are also expressed in gonadal cancers, but the data on their incidence in extra-gonadal tumors are scarce. Recently, the expression of FSHR in the vascular endothelium within different human cancers was found, but nothing is known on FSHR appearance in non-gonadal endocrine tumors. The present paper reports on the immunohistochemical detection of FSHR in human pituitary adenomas and adrenal tumors.</p><p><strong>Materials and methods</strong>. The study included samples of 28 pituitary adenomas and 36 adrenal tumors. Moreover, 2 samples of non-tumoral adrenal glands were also studied.</p><p>FSH receptor immunostaining was performed on paraffin sections using the rabbit anti-human FSH-R polyclonal antibody raised against 1-190 amino acid sequence from the human FSH-R (sc-13935). The pituitary adenomas were immunostained to reveal the pituitary hormones and the proliferation marker Ki-67.</p><p><strong>Results</strong>. In the pituitary adenomas, positive immunostaining with anti-FSHR antibody occurred in the adenoma cells cytoplasm and endothelia of the intra- and peritumoral blood vessels. The cytoplasmic immunostaining was found in the majority of investigated tumors but the intensity of staining was weak to moderate. There is some tendency towards the higher cytoplasmic FSHR score in tumors with higher Ki-67 index (atypical adenomas). In contrast to the cytoplasm, the FSHR immunostaining in blood vessels is strong and concerns all the investigated samples. Strong FSHR immunostaining is present in the endothelium of intra- and/or peritumoral blood vessels in the majority of pheochromocytomas, approximatively one half of the adrenocortical adenomas and both cases of the adrenal cancers. The immunostaining is detectable also in the tumoral cell cytoplasm in all but one examined pheochromocytomas.. All the investigated adrenocortical adenomas presented strong immunostaining of cell membranes. No immunostained cell membranes were found. in adrenal cancers. The positive immunostaining was found in glandular cells, but not in blood vessels, of non-tumoral adrenal cortex and medulla.</p><strong>Conclusions</strong>. Immunostaining of FSHR often occurs in the endothelium of intra- and/or peritumoral blood vessels of pituitary adenomas and benign and malignant adrenal tumors. The immunostaining may be also present in tumoral cells. A role of FSHR expression in these tumors (stimulation of angiogenesis? stimulation of cell growth?) needs further studies to be clarified

Immunohistochemical detection of FSH receptors in pituitary adenomas and adrenal tumors

Objectives. Follicle stimulating hormone (FSH) receptors (FSHR) are physiologically expressed in the ovary and testis. It is well known that FSHR are also expressed in gonadal cancers, but the data on their incidence in extra-gonadal tumors are scarce. Recently, the expression of FSHR in the vascular endothelium within different human cancers was found, but nothing is known on FSHR appearance in non-gonadal endocrine tumors. The present paper reports on the immunohistochemical detection of FSHR in human pituitary adenomas and adrenal tumors.Materials and methods. The study included samples of 28 pituitary adenomas and 36 adrenal tumors. Moreover, 2 samples of non-tumoral adrenal glands were also studied.FSH receptor immunostaining was performed on paraffin sections using the rabbit anti-human FSH-R polyclonal antibody raised against 1-190 amino acid sequence from the human FSH-R (sc-13935). The pituitary adenomas were immunostained to reveal the pituitary hormones and the proliferation marker Ki-67.Results. In the pituitary adenomas, positive immunostaining with anti-FSHR antibody occurred in the adenoma cells cytoplasm and endothelia of the intra- and peritumoral blood vessels. The cytoplasmic immunostaining was found in the majority of investigated tumors but the intensity of staining was weak to moderate. There is some tendency towards the higher cytoplasmic FSHR score in tumors with higher Ki-67 index (atypical adenomas). In contrast to the cytoplasm, the FSHR immunostaining in blood vessels is strong and concerns all the investigated samples. Strong FSHR immunostaining is present in the endothelium of intra- and/or peritumoral blood vessels in the majority of pheochromocytomas, approximatively one half of the adrenocortical adenomas and both cases of the adrenal cancers. The immunostaining is detectable also in the tumoral cell cytoplasm in all but one examined pheochromocytomas.. All the investigated adrenocortical adenomas presented strong immunostaining of cell membranes. No immunostained cell membranes were found. in adrenal cancers. The positive immunostaining was found in glandular cells, but not in blood vessels, of non-tumoral adrenal cortex and medulla.Conclusions. Immunostaining of FSHR often occurs in the endothelium of intra- and/or peritumoral blood vessels of pituitary adenomas and benign and malignant adrenal tumors. The immunostaining may be also present in tumoral cells. A role of FSHR expression in these tumors (stimulation of angiogenesis? stimulation of cell growth?) needs further studies to be clarified

Expression of somatostatin receptor subtypes in human thyroid tumors: the immunohistochemical and molecular biology (RT-PCR) investigation

Human endocrine tumors often express the somatostatin receptors SSTR 1–5 with different intensity. It has been widely investigated their distribution in pituitary adenomas, brain tumors, adrenal tumors and neuroendocrine tumors in gastrointestinal tract (NET). Some of studies also concern the expression of SSTRs in thyroid tumors but they are mainly limited to parafollicular C cells – derived medullary thyroid carcinomas (MTC). Results of SSTR 1–5 detection in other thyroid pathologies like follicular adenomas and papillary cancers are still scarce and often controversial, depending of investigation method used. The aim of this study was to report the presence of all the 5 subtypes of SSTR (including 2A and 2B SSTR isoforms) in some surgically treated human thyroid tumors by means of immunohistochemistry and real-time PCR method and to correlate the results obtained with both techniques. SSTR 1 protein was expressed in 88.8% of investigated cases, SSTR 2A and 2B both in 44.4%, SSTR 3 in 55.5%, SSTR 4 in 11.2% and SSTR 5 in 33.3%. SSTR 1 is the dominant form in the thyroid gland tumor and hyperplasia. We found positive confirmation of both methods in 88.8% for SSTR 1, 2A, 3 subtypes, in 22.2% for SSTR 4 and in 100% for SSTR 5. It suggests that somatostatin multiligand analogs or selective SSTR 1 agonists may be used in thyroid tumors treatment

Stiffness memory of EA.hy926 endothelial cells in response to chronic hyperglycemia

Background: Glycemic memory of endothelial cells is an effect of long-lasting hyperglycemia and is a cause of various diabetics complications, that arises despite of the treatment targeted towards returning low glucose level in blood system. On the other hand, endothelial dysfunction, which is believed to be a main cause of cardiovascular complications, is exhibited in the changes of mechanical properties of cells. Although formation of the glycemic memory was widely investigated, its impact on the mechanical properties of endothelial cells has not been studied yet. Methods: In this study, nanoindentaion with a tip of an atomic force microscope was used to probe the long-term changes (through 26 passages, c.a. 80 days) in mechanical properties of EA.hy926 endothelial cells cultured in hyperglycemic conditions. As a complementary method, alterations in the structure of actin cytoskeleton were visualized by fluorescent staining of F-actin. Results: We observed a gradual stiffening of the cells up to 20th passage for cells cultured in high glucose (25 mM). Fluorescence imaging has revealed that this behavior resulted from systematic remodeling of the actin cytoskeleton. In further passages, a drop in stiffness had occurred. The most interesting finding was recorded for cells transferred after 14 passages from high glucose to normal glucose conditions (5mM). After the transfer, the initial drop in stiffness was followed by a return of the cell stiffness to the value previously observed for cells cultured constantly in high glucose. Conclusions: Our results indicate that glycemic memory causes irreversible changes in stiffness of endothelial cells. The formation of the observed "stiffness memory" could be important in the context of vascular complications which develop despite the normalization of the glucose level

Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry

The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found