Hormones and depression in the advanced age.

INTRODUCTION: In older people, depression and mental disability are more frequently present than in younger subjects. It is found that depressionis a risk factor for dementia, just as dementia is a risk factor for depression. In turn, both disturbances are known to be influenced by hormones. The present study aimed to see whether the hormonal changes in subjects over 75 years old correlate with the symptoms of depression measured using the Geriatric Depression Scale (GDS). MATERIAL AND METHODS: In a group of patients aged over 75 years, concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol, testosterone, dehydroepiandrosterone sulphate (DHEAs), and cortisol were measured in serum with the use of chemiluminescence. The symptoms of depression were estimated by GDS, and the mental functions were assessed by the Mini-Mental State Examination (MMSE). The correlations between the obtained results were estimated by Spearman's test. RESULTS AND CONCLUSIONS: A significant correlation between GDS and MMSE scores was observed in the investigated patients. Some statistically significant correlations concerning cortisol and testosterone with GDS were observed in women, but not in men. On the other hand, no significant correlations between concentrations of FSH, LH, DHEAs, and oestradiol with GDS were noticed. Our data support the role of cortisol (possibly secreted during chronic stress) in the risk of depression. The gender difference in the mechanism of depression and stress in older age could be also hypothesized.

Serum testosterone level correlates with left ventricular hypertrophy in older women.

Introduction: Sex hormones may play an important role in age-related cardiac remodeling. However, their impact on cardiac structure and function in females of advanced age still remains unclear. The aim of this study is to evaluate the relationship between sex hormones level and echocardiographic parameters in older women with concomitant cardiovascular diseases. Materials and Methods: The study group included 52 community-dwelling women with mean age 79.5 ± 2.8 years, consecutive patients of an outpatient geriatric clinic. In all the subjects, a transthoracic echocardiogram was performed and serum testosterone, estradiol, follicle-stimulating hormone, luteinising hormone, dehydroepiandrosterone sulphate, and cortisol levels were determined. Results: Testosterone level correlated positively with interventricular septum diastolic dimension (IVSd) (rS=0.293, p<0.05), left ventricular mass index (rS=0.285, p<0.05), E/E' ratio (rS=0.301, p<0.05), and negatively with E' (rS=-0.301, p<0.05). Estradiol level showed a positive correlation with the posterior wall dimension (rS=0.28, p<0.05). Besides, no significant correlations between clinical or echocardiographic parameters and other hormones were observed. Female subjects with diagnosed left ventricular hypertrophy (LVH) (n=34) were characterized by a significantly higher rate of hypertension (p=0.011), higher waist-to-height ratio (p=0.009), higher testosterone level (0.82 vs. 0.48 nmol/L, p=0.024), higher testosterone/estradiol ratio (16.4 vs. 9.9, p=0.021), and received more anti-hypertensive drugs (p=0.030). In a multiple stepwise logistic regression, the best determinants of LVH were the presence of hypertension (OR=6.51; 95% CI 1.62-26.1), and testosterone level (OR= 6.6; 95% CI 1.19-36.6). Conclusions: Higher serum testosterone levels may contribute to pathological cardiac remodeling, especially in hypertensive women. Estradiol, gonadotropins, DHEAS, and cortisol were not related to echocardiographic parameters.

Gonadotropins at Advanced Age - Perhaps They Are Not So Bad? Correlations Between Gonadotropins and Sarcopenia Indicators in Older Adults.

Many hormones fluctuate during the aging process. It has been suggested that gonadotropins, which increase with age, contribute to the occurrence of many diseases and syndromes in older life, such as cardiovascular diseases, obesity, frailty syndrome and osteoporosis. This study aims to assess the relationship between circulating gonadotropins and other hormones potentially contributing to age-related functional decline and sarcopenia indicators in 39 male and 61 female community-dwelling seniors, mean age 80 years. According to the definition developed by the second European Working Group on Sarcopenia in Older People (EWGSOP2), the following indicators of the sarcopenia were assessed: bioimpedance-measured body composition, gait speed, handgrip strength, timed up and go test (TUG), chair stand test, Short Physical Performance Battery (SPPB). Blood levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, testosterone, dehydroepiandrosterone sulphate (DHEAS) and cortisol were also measured. In the men, FSH and partially LH correlated positively with muscle mass percentage, gait speed, handgrip strength and SPPB, and negatively with percent body fat. Additionally, testosterone and DHEAS correlated negatively with the percentage of fat mass in men. Whereas in the women, FSH and LH were mainly negatively associated with body mass and adipose tissue measures. Cortisol did not show any relationship with the examined indicators. The study shows that the indicators of sarcopenia are strongly associated with levels of gonadotropins, sex hormones and DHEAS, especially in older men. The obtained results, after being confirmed in a larger group, may modify prevention and treatment strategies of sarcopenia.

Are physical and mental abilities of older people related to gonadotropins and steroid hormones levels?

BACKGROUND: Aging is characterized by deep alterations of hormone secretion. In majority, hormone secretion, except gonadotropins, undergoes a pronounced decrease which is thought to contribute to the progression of aging. The recent data indicate that gonadotropin excess may also by itself influence the aging process. The aim of the present study was to investigate the relations between gonadotropins and steroid hormones with physical and mental abilities of older people. MATERIAL AND METHODS: In a group of patients aged over 75 years, concentrations of FSH, LH, estradiol, testosterone, DHEAs and cortisol were measured. The mental ability was estimated by MMSE and CDT and the physical ability by TUG and SPPB tests. RESULTS AND CONCLUSIONS: The positive correlation between SPPB scores and FSH and the negative correlations of SPPB with LH/FSH ratio were observed in men. The correlation of TUG scores and estradiol levels was also noted in men. The positive correlation between CDT scores and FSH in women and the negative correlation between CDT and LH/FSH ratio in men were found. The correlation between the results of CDT and cortisol levels in men was also observed. Thus, we did not confirm the simple deleterious effect of gonadotropins on cognitive abilities. FSH and LH seem exert different (antagonistic?) effects on cognitive functions, but this hypothesis needs further studies.

Gonadotropins and steroid hormones in older people: their mutual connections and relations to body mass indices.

INTRODUCTION: Aging in mammals, including man, is accompanied by deep changes in hormone secretion. In the majority of cases, hormone secretion (mostly of gonadal steroids and adrenocortical hormone dehydroepiandrosterone - DHEA) undergoes pronounced decrease. This decrease is thought to contribute to the progression of aging. In contrast, the secretion of gonadotropins is sharply increased in older adults, as a result of gonadal deficiency. Recent data indicate that gonadotropin excess may also, by itself, influence the aging process. The aim of the present study was to investigate the mutual relation between steroid hormones and gonadotropins and their effect on body mass indices in older people. MATERIAL AND METHODS: In a group of 100 patients (61 women and 39 men) aged over 75 years, blood serum concentrations of folliclestimulating hormone (FSH), luteinising hormone (LH), oestradiol (E2), testosterone, dehydroepiandrosterone sulphate (DHEAs), and cortisol were measured. All the patients were measured for the following: body weight (kg), body mass index (BMI) (kg/m²), and waist-to-height ratio (WtHR). The differences of the numerical data were evaluated by Student's t-test and the correlations between them by means of Pearson's test. RESULTS AND CONCLUSIONS: The most interesting finding of this study was to show that FSH and LH are negatively correlated with body mass and indices such as BMI and WtHR in older women. Because in older women the mediation of ovary is unlikely, we conclude that gonadotropins may influence the body mass by their direct extra-gonadal action.

Expression of α-Klotho protein in human thyroid cancers - an immunohistochemical study.

INTRODUCTION: The α-Klotho protein was discovered as a gene controlling the process of aging, but further studies indicated that it also plays the role of a tumour suppressor. Although numerous studies were performed on the role of the α-Klotho gene and protein in neoplasia, the data on α-Klotho protein expression in thyroid cancers are very scarce. Our study presents the immunohistochemical investigation of α-Klotho expression in benign and malignant thyroid tumours. MATERIAL AND METHODS: The material included samples of benign (nodular hyperplasia, follicular adenoma), differentiated (follicular and papillary) cancers and aggressive thyroid cancers of low differentiation grade. The samples were immunostained using two different monoclonal anti-α-Klotho antibodies. RESULTS: From the two antibodies used in this study, one (EPR6856) reacted probably with the soluble form of Klotho and immunostained mostly the colloid filling thyroid follicles and intravascular or extravascular serum deposits. The other (A-9 antibody) immunostained the follicular epithelium in benign thyroid lesions as well as the epithelial tumoural cells in differentiated thyroid (follicular and papillary cancers). In the thyroid cancers of high malignancy, the immunostaining with A-9 anti-α-Klotho antibody was (except in one case) negative or very weak. CONCLUSION: Our results indicate that lowered expression of a- Klotho is involved in the process of thyroid neoplasia.

"Silent" thyrotropin (TSH) expression in acromegaly and clinically non-functioning pituitary adenomas.

INTRODUCTION: The pituitary adenomas secreting thyrotropin (TSH) are considered the rarest type of hormonally active pituitary tumour. In spite of that, many cases are described in the literature. On the other hand, the observations of the co-expression of TSH with other pituitary hormones (mostly with growth hormone [GH]) and "silent" expression of TSH in clinically non-functioning pituitary adenomas (CNFPA) are rather scarce. MATERIAL AND METHODS: Among 93 examined pituitary adenomas, 22 of them were diagnosed as active acromegaly and 71 as clinically non-functioning pituitary adenomas (CNFPA). All of them were immunostained with antibodies against pituitary hormones, including the anti-TSH antibody. TSH-immunopositive adenomas are immunostained also to detect somatostatin receptor subtypes (SSTR 1-5). RESULTS: TSH immunopositivity was found in 4.2% of CNFPA (3/71 tumours) and in 13.6% (3/22) cases of somatotropinomas manifesting as active acromegaly. All of the examined TSH-immunopositive adenomas expressed SSTR subtypes except SSTR4. The symptoms of hyperthyroidism were not observed in any of the acromegalic patients co-expressing TSH with GH. CONCLUSIONS: Our data confirm the relative rarity of TSH expression or co-expression of TSH in pituitary tumours. In most cases TSH is co-expressed with GH in patients with acromegaly and is not accompanied by hyperthyroidism. The "silent" expression of TSH may occur also, although rarely in CNFPA. The strong expression of SSTR in TSH-immunopositive CNFPA ("silent thyrotropinoma") indicates the possibility of the treatment of these tumours with somatostatin analogues. (Endokrynol Pol 2016; 67 (5): 515-518).

An evaluation of the effects of somatostatin analogue therapy in non-functioning pituitary adenomas in comparison to acromegaly.

INTRODUCTION: Non-functioning pituitary adenomas (NFPA) are often diagnosed late as invasive macroadenomas. The surgical resection is usually incomplete and about 50% of patients require additional surgery. Recent data suggest that somatostatin analogues (SSA), so important in the pharmacotherapy of acromegaly, can also be effective in the management of NFPA. MATERIAL AND METHODS: We analysed data of patients who had been treated up to 10 years previously with SSA: 40 with acromegaly (23 - primary, 17 - recurrent tumours) and 22 with NFPA (4 - primary, 18 - recurrent tumours). Hormonal profile, dynamics of tumour size change, ophthalmic syndromes, somatostatin receptor (SSTR) scintigraphy, and immunohistochemistry of SSTR subtypes of operated tumours as well as side effects were investigated. RESULTS: Biochemical cure of acromegaly was achieved in 57.5% of patients, while reduction of tumour size was observed in 37% of patients and it was more frequent in not-operated cases. Regarding NFPA, stabilisation of tumour size was noticed in 68% of patients. Tumour shrinkage was reported in 9% of cases, but in 23% of the study group the adenoma size increased with indication for reoperation. CONCLUSIONS: The efficacy of SSA in NFPA is much lower in comparison to their well-established effects in the treatment of acromegaly. Stabilisation of tumour size, which is observed in the majority of NFPA, is significantly more frequent in comparison to the natural history of untreated NFPA and our previous studies as well. Analysis of SSTR subtypes is an argument in favour of introduction of novel broad-spectrum SSA that may be more effective in the treatment of NFPA. Referring to acromegaly, adenoma size decrease was reported more frequently in primary therapy. Considering recurrent tumours better outcomes were achieved in patients who were pre-treated with SSA before planned surgery. (Endokrynol Pol 2016; 67 (3): 292-298).

Ultrafiltration - an alternative method to polyethylene glycol precipitation for macroprolactin detection.

INTRODUCTION: The aim of the study was to evaluate two methods of macroprolactin (MaPRL) detection - precipitation with polyethylene glycol (PEG) and ultrafiltration and to compare these techniques with "gold standard" - gel filtration chromatography (GFC). MATERIAL AND METHODS: The study was conducted on 245 patients - 45 with organic and 200 with functional hyperprolactinaemia. In all the subjects MaPRL was detected by precipitation with PEG and ultrafiltration. Additionally, gel filtration chromatography was performed in some of the serum samples. RESULTS: Macroprolactinaemia was detected in 27 patients - 8 with prolactinoma and 19 with functional hyperprolactinaemia. Assessing positive and negative results for MaPRL, we observed high diagnostic agreement (95.9%) and positive correlation (r = 0.506, p < 0.001) between the methods. The results of precipitation and ultrafiltration positive for MaPRL were concordant in 63%. The dominance of MaPRL detected with precipitation and/or ultrafiltration was confirmed by GFC in 76% of cases (all patients with functional hyperprolactinaemia). Among 6 examined patients with prolactinoma, GFC showed four false-positive results - 1 case of precipitation and 3 cases of ultrafiltration. CONCLUSIONS: Efficacy of MaPRL detection with precipitation and ultrafiltration is comparable especially in cases of functional hyperprolactinaemia. In patients with prolactinoma, precipitation seems to be a more efficient separation method.

Expression of follicle stimulating hormone receptors (FSHR) in thyroid tumours - a marker of malignancy?

BACKGROUND: In normal conditions FSHR are expressed in granulosa cells of the ovary and Sertoli cells of the testis. They can be expressed also in gonadal tumours. However, recently the expression of FSHR was found in tumoral cells and intra-tumoral blood vessels of many other tumours, including thyroid tumours. Aim of this study was to see whether the expression of FSHR can be useful in the differentiation of benign and malignant thyroid lesions. METHODS: 44 samples of surgically excised thyroids were immunostained with anti- FSHR antibody raised against 1-190 amino acid sequence from the human FSHR. RESULTS: Non-neoplastic thyroid follicles (i.e. the follicles situated outside the tumour) do not show the immunostaining for FSHR. The same concerns the majority of follicular adenomas. In contrast, 87.5% of follicular cancers, the same percentage of papillary cancers and all the examined undifferentiated cancers showed the FSHR immunopositivity of tumoral cells. A tendency towards the higher frequency of FSHR - positive blood vessels also concerns malignant thyroid tumours. CONCLUSIONS: The ectopic FSHR immunostaining seems to be useful to differentiate malignant from benign lesions, especially follicular cancers from follicular adenomas. However, the further studies on larger material are needed.

Ectopic expression of follicle-stimulating hormone receptors in thyroid tumors.

INTRODUCTION: In normal conditions follicle-stimulating hormone receptors (FSHR) are expressed in the ovary and the testis. They can also be expressed in gonadal tumors. However, recently we have found FSHR immunostaining in pituitary adenomas, adrenal tumors and neuroendocrine tumors (carcinoids). The aim of this study was to determine whether the same occurs in thyroid tumors. MATERIAL AND METHODS: Thirty-six samples of surgically excised thyroids were examined. Follicle-stimulating hormone receptors immunostaining was performed on paraffin sections using the rabbit anti-human FSHR polyclonal antibody raised against a 1-190 amino acid sequence from the human FSHR (sc-13935, Santa Cruz). RESULTS: Normal thyroid follicles do not show immunopositivity for FSHR. The same concerns the majority of benign lesions, diagnosed as hyperplasia nodularis or thyroid adenomas. However, positive FSHR immunostaining in some follicles was observed. In all but one thyroid cancer (15 papillary, 10 follicular cancers and one case of anaplastic thyroid cancer) 10-100% of tumor cells exhibit positive FSHR immunostaining. In about 40% of samples FSHR immunoreactivity can be observed also in the endothelia of intrathyroidal blood vessels. This immunopositivity was more frequent in the samples of thyroid cancers (13/27) than in benign lesions (2/9). CONCLUSIONS: Ectopic positive FSHR immunostaining is also present in thyroid cancers, and, to a lesser degree, in benign lesions but not in the normal thyroid epithelium.

Usefulness of PTH measurements in FNAB washouts in the identification of pathological parathyroids--analysis of the factors that influence the effectiveness of this method.

INTRODUCTION: The aim of this investigation was to assess the usefulness of the measurement of PTH concentration in the material obtained during FNAB (PTH-FNAB) in the identification of pathological parathyroids in patients with frequently coexisting thyroid abnormalities (nodular goitre, chronic thyroiditis, previous thyroidectomy). Additionally, the influence of the size of goitre, parathyroid localisation and size on the results of PTH-FNAB measurement was examined. MATERIAL AND METHODS: Fifty patients with primary hyperparathyroidism and sonographically detected focal lesion that was suggestive of parathyroid gland were included in this study. PTH-FNAB results were correlated with the outcome of routine cytological examination and biochemical indices of hyperparathyroidism, SPECT-CT (33 patients) and histopathological examination (20 patients). RESULTS: Positive PTH-FNAB was observed in 80% of patients, and in more than 70% of persons with non-diagnostic smears or smears 'contaminated' with thyroid follicular cells. In the group of operated patients, sensitivity of PTH-FNAB (95.0%) was higher than SPECTCT (64.3%, p < 0.05). Presence of nodular goitre and/or chronic thyroiditis exerts a two times stronger negative effect on percentage of negative results of SPECT-CT than of PTH-FNAB. On the other hand, lower frequency of positive PTH-FNAB but not SPECT-CT was observed when the thickness of the thyroid was ≥ 20 mm (50% v. 87.5%, p < 0.05) and when the thickness of a lesion suspected of parathyroid pathology was ≤ 5 mm (66.7% v. 93.3%, p < 0.05). CONCLUSIONS: In patients with thyroid abnormalities, PTH-FNAB measurements show advantages over routine biopsy and SPECT-CT in the identification of typically located pathological parathyroids.

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.

Ghrelin concentration is correlated with IGF-I/IGFBP-3 molar ratio but not with GH secretion in children with short stature.

OBJECTIVES: In children with growth hormone deficiency (GHD) and neurosecretory dysfunction (NSD) ghrelin concentrations are significantly higher than in children with idiopathic short stature (ISS), however the correlation between serum ghrelin and growth hormone (GH) is not observed. The aim of the study was to compare ghrelin concentrations with IGF-I/IGFBP3 molar ratio in children with short stature due to different etiology. MATERIAL: Analysis comprised 136 children (58 girls and 78 boys), aged 3.86-16.82 years with short stature (below -2.0 SD); in 21 of them GHD was diagnosed, in 23 - NSD and 92 - ISS. In each child, fasting ghrelin, insulin-like growth factor type I (IGF-I) and its binding protein type 3 (IGFBP-3) concentrations were measured. The results were analysed separately in younger and in older children. Depending on IGF-I/IGFBP-3 molar ratio, children were divided into two (2) groups: with lower IGF-I/IGFBP-3 and with higher IGF-I/IGFBP-3 ratio value. RESULTS: Both in younger and in the older age groups, ghrelin concentration was significantly higher in children with lower IGF-I/IGFBP-3 ratio than in children with higher IGF-I/IGFBP-3 value (1937.3±1232.4 vs 1365.3±632.1 pg/ml in younger children and 1205.4±548.8 vs 867.4±282.9 pg/ml in older children). The negative correlation between ghrelin and IGF-I/IGFBP-3 ratio was observed in both age groups. Not only children with GHD and NSD, but also as much as 39% out of all children with ISS were qualified into the subgroups with lower IGF-I/IGFBP-3 ratio. CONCLUSIONS: Ghrelin secretion is elevated in children with lower IGF-I/IGFBP-3 ratio. It seems that lower bioactivity of IGF-I is stimulating factor for ghrelin synthesis.

Effects of nitric oxide synthase inhibition on diethylstilbestrol-induced hyperprolactinaemia and pituitary tumourigenesis in rats.

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.

The detection of macroprolactin by precipitation and ultrafiltration methods.

BACKGROUND: Prolactin (PRL) exists in human blood in several molecular forms. Macroprolactin (MaPRL), which most often consists of monomeric PRL and immunoglobulin G, has the highest molecular weight but no biological activity. Immunoassays do not distinguish MaPRL from monomeric PRL, what can lead to an incorrect diagnosis of hyperprolactinaemia. The most commonly used technique to separate the isoforms of PRL is precipitation with polyethylene glycol (PEG). Another technique - ultrafiltration - seems to be useful in MaPRL detection. The aim of this study was to evaluate the occurrence of MaPRL in hyperprolactinaemic patients and to compare the results obtained by precipitation and ultrafiltration. MATERIAL AND METHODS: The study was conducted on 120 sera obtained from patients hospitalised in the Department of Clinical Endocrinology, Medical University of Lodz, in whom PRL concentration was above 30 ng/mL Of these 120 patients, 25 had pituitary adenoma, 52 had polycystic ovary syndrome (PCOS), and 43 had idiopathic hyperprolactinaemia (HPRL). Macroprolactin was detected using two methods: precipitation with PEG and ultrafiltration. Concentration of PRL was measured by Immulite 1000 immunoassay (Siemens). RESULTS: We detected a predominance of MaPRL in ten patients (three with macroprolactinoma, three with PCOS and four with HPRL) using precipitation and ultrafiltration. Positive correlation and diagnostic concordance between the results of precipitation and ultrafiltration were noted, especially in the group with functional hyperprolactinaemia. In half of the patients with macroprolactinaemia, and in 12 of the 110 subjects without significant amounts of MaPRL, real PRL concentration was within the reference range. CONCLUSIONS: MaPRL is not a significant clinical problem in the studied population. However, in patients with hyperprolactinaemia, especially non-organic, screening for macroprolactinaemia should be performed. The effectiveness of the precipitation and ultrafiltration methods for detecting MaPRL is comparable in functional hyperprolactinaemia, but the usefulness of ultrafiltration in patients with pituitary adenoma requires further examination.

Immunohistochemical detection of dopamine D2 receptors in neuroendocrine tumours.

BACKGROUND: Recently, dopamine D2 receptors (RD2) have been found to be expressed in neuroendocrine tumours (NET), the tumours which arise from the diffuse neuroendocrine cells. Moreover, successful trials of the treatment of NET with cabergoline - D2 agonist, have been reported. These findings increase the interest of investigating RD2 expression in NET. MATERIAL AND METHODS: The expression of RD2 was investigated immunohistochemically using the antibody which recognises both short (S) and long (L) isoforms of the receptor in 17 NET samples taken from 15 patients. RESULTS: In 17 NET samples, a positive reaction with the anti-RD2 antibody occurred in 11 cases. In six cases, the localisation of the immunostaining was cytoplasmic and in nine cases it was nuclear. Only in one case was the receptor cell membrane-located, and in two cases the immunoreaction was also localised in the blood vessels walls. The relation between RD2 expression and the grade of malignancy examined by means of Ki-67 antigen expression needs further study. However, preliminary observations indicate that the nuclear localisation of RD2 is linked to higher tumour malignancy. The next investigated question was the co-expression of somatostatin and dopamine receptors. This question seems important because of the perspectives of somatostatin-dopamine chimeras application in NET treatment. In the samples examined by us, RD2 were co-expressed in 5/10 cases with sstr1, in 3/10 with sstr2A, in 2/9 with sstr2B, in 3/10 with sstr3, and in 5/10 with sstr5. CONCLUSION: Dopamine D2 receptors are revealed by means of immunohistochemistry in the majority of NET. They exhibit cytoplasmic and/or nuclear localisations, the latter being possibly linked to a higher grade of malignancy, and are often co-expressed with somatostatin receptors (mostly with subtypes1 and 5).

Differential expression of somatostatin receptor subtype-related genes and proteins in non-functioning and functioning adrenal cortex adenomas.

Adrenocortical adenomas display highly variable expressions of somatostatin receptor (SSTR) subtypes, whose expression is mandatory (although not always sufficient) to achieve the positive effects of somatostatin (SST) analog therapy. Immunohistochemistry (IHC) is the main method used to investigate receptor protein expression. The molecular biology method - polymerase chain reaction (PCR) - is also often used to investigate receptor expression. Nevertheless, the expression of receptor mRNA and the respective receptor protein is not always synchronized. The aim of this study was to investigate SSTR expression by IHC in adrenal adenomas, to compare the results to data obtained by real-time PCR and to determine whether hormonally functioning and non-functioning adenomas differ in this respect. Adrenocortical adenomas were removed surgically from 13 females and 2 males. The tissues were obtained from 9 non-functioning and 6 functioning adenomas. The intensity of IHC reaction was scored semiquantitatively by two independent observers. Real-time PCR was performed using pairs of primers in a reaction amplified along a gradient of temperatures. Amplified DNA was measured by monitoring SYBR-Green fluorescence. In non-functioning tumors, compatibility between IHC and PCR results was observed for SSTR 1 and 2 in 62.5% of the samples. Fifty percent of patients demonstrated compatibility for SSTR 4 and 5 and 37.5% for SSTR 3. In hormonally active adenomas, total compatibility of both methods was noted for SSTR 2 (100%). The compatibility obtained for SSTR 5 was 66.6%. We conclude that receptor gene and respective receptor protein expression are not always synchronized. Messenger RNA detection alone is not sufficient to predict the presence of the receptor protein acting as a target for SST and its analogs.

Expression of somatostatin receptor subtypes in primary and recurrent gonadotropinomas: are somatostatin receptors involved in pituitary adenoma recurrence?

OBJECTIVE: Surgical treatment of pituitary macroadenomas often fails because of tumor recurrence after the operation. The causes of tumor recurrence are complex, but one of them may be the high growth potential of the adenoma. As somatostatin receptors mediate antiproliferative, anti-angiogenic and pro-apoptotic actions, it seemed reasonable to investigate their expression in dependence on the adenoma recurrence. METHODS: Samples of primary and recurrent gonadotropinomas excised surgically from patients were examined. This type of pituitary adenomas was chosen because of its relatively high recurrence rate. The adenoma phenotype and expression of somatostatin receptor subtypes 1-5 (SSTR 1-5) were investigated by immunohistochemistry, and the level of SSTR expression was semiquantitatively scored. RESULTS: It was found that the adenomas undergoing the early recurrence have lower expression of SSTR 2A and 3 in comparison to those which did not recur during 5 years lasting observation. On the other hand, the recurrent tumors show higher expression of SSTR 1, 2A, 3 and 5 subtypes than their primary counterparts. CONCLUSIONS: It is hypothesized that SSTR may, at least in part, counteract adenoma recurrence. On the other hand, it can be also presumed that the recurrent gonadotropinomas may be more sensitive to somatostatin analog treatment than primary ones. These hypotheses need to be confirmed in further studies.

SSTR1 and SSTR5 subtypes are the dominant forms of somatostatin receptor in neuroendocrine tumors.

The effectiveness of the long acting somatostatin analogues like octreotide and lanreotide depends on the expression of specific somatostatin receptors on the target cells. The immunohistochemical method performed on surgically removed tumors searches the expression of receptors at the level of receptor protein and gives us insight into receptor's cellular localization. The aim of study was to assess the presence of all the 5 subtypes of SSTR 1-5 (including 2A and 2B SSTR isoforms) in surgically treated human neuroendocrine tumors (NETs) to establish which receptor subtype is the dominant form of somatostatin receptor in particular tumor and thus to be able to predict which somatostatin analog will be effective in NETs treatment. 18 samples of neuroendocrine tumors (surgically excised tumors or biopsies) were immunostained with specific antibodies. Expression of SSTR was scored semiquantitatively. Only strong or moderate immunostaining was considered as positive reaction. The summarized expression pattern of SSTR in the investigated neuroendocrine tumors in our material was: SSTR 1> SSTR 5> SSTR 3> SSTR 2A> SSTR 2B. The receptors were distributed mainly in the area of cells cytoplasm with a few specimens showing only membranous or mixed: membranous--cytoplasmic localization. The observed pattern suggests that apart from octreotide and lanreotide, newly synthesized multiligand analogs such as SOM 230, KE 108 or SSTR 1 and SSTR 5 selective analogs could be effective in NETs treatment.