Monitoring psychoactive substance use at six European festivals through wastewater and pooled urine analysis.

The consumption of psychoactive substances is considered a growing problem in many communities. Moreover, new psychoactive substances (NPS) designed as (legal) substitutes to traditional illicit drugs are relatively easily available to the public through e-commerce and retail shops, but there is little knowledge regarding the extent and actual use of these substances. This study aims to gain new and complementary information on NPS and traditional illicit drug use at six music festivals across Europe by investigating wastewater and pooled urine. Samples were collected, between 2015 and 2018, at six music festivals across Europe with approximately 465.000 attendees. Wastewater samples were also collected during a period not coinciding with festivals. A wide-scope screening for 197 NPS, six illicit drugs and known metabolites was applied using different chromatography-mass spectrometric strategies. Several illicit drugs and in total 21 different NPS, mainly synthetic cathinones, phenethylamines and tryptamines, were identified in the samples. Ketamine and the traditional illicit drugs, such as amphetamine-type stimulants, cannabis and cocaine were most abundant and/or frequently detected in the samples collected, suggesting a higher use compared to NPS. The analyses of urine and wastewater is quick and a high number of attendees may be monitored anonymously by analysing only a few samples which allows identifying the local profiles of use of different drugs within a wide panel of psychoactive substances. This approach contributes to the development of an efficient surveillance system which can provide timely insight in the trends of NPS and illicit drugs use.

A20/TNFAIP3 heterozygosity predisposes to behavioral symptoms in a mouse model for neuropsychiatric lupus.

Background: Neuropsychiatric lupus (NPSLE) refers to the neurological and psychiatric manifestations that are commonly observed in patients with systemic lupus erythematosus (SLE). An important question regarding the pathogenesis of NPSLE is whether the symptoms are caused primarily by CNS-intrinsic mechanisms or develop as a consequence of systemic autoimmunity. Currently used spontaneous mouse models for SLE have already contributed significantly to unraveling how systemic immunity affects the CNS. However, they are less suited when interested in CNS primary mechanisms. In addition, none of these models are based on genes that are associated with SLE. In this study, we evaluate the influence of A20, a well-known susceptibility locus for SLE, on behavior and CNS-associated changes in inflammatory markers. Furthermore, given the importance of environmental triggers for disease onset and progression, the influence of an acute immunological challenge was evaluated. Methods: Female and male A20 heterozygous mice (A20+/-) and wildtype littermates were tested in an extensive behavioral battery. This was done at the age of 10±2weeks and 24 â€‹± â€‹2 weeks to evaluate the impact of aging. To investigate the contribution of an acute immunological challenge, LPS was injected intracerebroventricularly at the age of 10±2weeks followed by behavioral analysis. Underlying molecular mechanisms were evaluated in gene expression assays on hippocampus and cortex. White blood cell count and blood-brain barrier permeability were analyzed to determine whether peripheral inflammation is a relevant factor. Results: A20 heterozygosity predisposes to cognitive symptoms that were observed at the age of 10 â€‹± â€‹2 weeks and 24 â€‹± â€‹2 weeks. Young A20+/- males and females showed a subtle cognitive phenotype (10±2weeks) with distinct neuroinflammatory phenotypes. Aging was associated with clear neuroinflammation in female A20+/- mice only. The genetic predisposition in combination with an environmental stimulus exacerbates the behavioral impairments related to anxiety, cognitive dysfunction and sensorimotor gating. This was predominantly observed in females. Furthermore, signs of neuroinflammation were solely observed in female A20+/- mice. All above observations were made in the absence of peripheral inflammation and of changes in blood-brain barrier permeability, thus consistent with the CNS-primary hypothesis. Conclusions: We show that A20 heterozygosity is a predisposing factor for NPSLE. Further mechanistic insight and possible therapeutic interventions can be studied in this mouse model that recapitulates several key hallmarks of the disease.

Towards antimicrobial yet bioactive Cu-alginate hydrogels.

The simplest approach to enhance alginate hydrogel characteristics and functional properties is to replace the calcium in the process of alginate gelation with other metallic ions which are essential for living systems. Gelling of alginate with other ions and using modern encapsulation techniques can provide new delivery systems with required properties. Hence, in this study Cu-alginate hydrogels in the form of microbeads were produced by electrostatic extrusion using gelling solutions with Cu(II) concentrations in the range 13.5-270 mM and comprehensively characterized in vitro. The variation of gelling solution concentration influenced the microbead Cu(II) content, size, biomechanical properties, Cu(II) release and subsequently potential biomedical application. The formulations chosen for biomedical evaluation showed potential for antimicrobial and tissue engineering applications. Microbeads with higher Cu(II) loading (~100 µmol g(-1)) induced immediate bactericidal effects against Escherichia coli and Staphylococcus aureus. Conversely, Cu(II) release from microbeads with the Cu(II) content of ~60 µmol g(-1) was slower and they were suitable for promoting and maintaining chondrogenic phenotype of bovine calf chondrocytes in 3D culture. Results of this study have shown possibilities for tuning Cu-alginate properties for potential biomedical applications such as antimicrobial wound dressings, tissue engineering scaffolds or articular cartilage implants.

Daidzein administration positively affects thyroid C cells and bone structure in orchidectomized middle-aged rats.

SUMMARY: Thyroid C cells hormone, calcitonine, inhibits bone resorption. We have demonstrated that daidzein treatment of orchidectomized rats (model for osteoporosis) stimulated C cells and increased trabecular bone mass. These results suggest that, besides direct action, daidzein may also affect bone structure indirectly through enhancement of thyroid C cell activity. INTRODUCTION: Thyroid C cells produce calcitonin (CT) which acts as an inhibitor of bone resorption. In this study, the influence of daidzein treatment on thyroid C cells, bone structure, and bone function in orchidectomized (Orx) middle-aged rats was investigated. METHODS: Sixteen-month-old Wistar rats were divided into Orx and sham-operated (SO) groups. Half the Orx rats were given subcutaneous injections of daidzein (30 mg/kg b.w./day) for 3 weeks. CT-immunopositive thyroid C cells were morphometrically analyzed. The metaphyseal region of the proximal tibia was measured histomorphometrically, and cancellous bone area (B.Ar), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) were calculated. Serum samples were analyzed for CT and osteocalcin (OC), calcium (Ca) and phosphorus concentrations, and urine samples for Ca levels. RESULTS: Treatment of Orx animals with daidzein significantly increased volume of C cells compared to the Orx rats. Daidzein also enhanced B.Ar, Tb.Th, and Tb.N and reduced Tb.Sp. The serum OC and urinary Ca concentrations decreased significantly in comparison with the Orx group. CONCLUSIONS: These findings indicate that daidzein treatment stimulates thyroid C cells, increase trabecular bone mass, and decrease bone turnover in Orx middle-aged rats, which is the model of male osteoporosis.

Maternal dexamethasone treatment reduces ovarian follicle number in neonatal rat offspring.

Elevated glucocorticoid levels in the gravid female circulation affect a number of endocrine functions in the fetuses and neonates. The aim of this study was to examine the effects of maternal dexamethasone (Dx) administration during late pregnancy on the ovaries of neonatal offspring. On the 16th day of pregnancy, experimental dams received subcutaneously 1.0 mg Dx/kg b.w., followed by 0.5 mg Dx/kg b.w./day on the 17th and 18th days of gestation. The control gravid females received the same volume of saline vehicle. Left ovaries from 5-day-old female pups were stereologically analyzed. The ovary volumes were estimated using Cavalieri's principle. The number of healthy and atretic primordial and primary follicles was estimated using a fractionator-physical dissector method. The number of secondary follicles was determined by exact counts of every fourth section encompassing whole cross-sections of the ovary. The ovary volume was significantly decreased (by 44.4%; P < 0.05) in the group of female pups from Dx-treated mothers comparing to the controls. The numbers of healthy primordial and atretic follicles were 38.8% (P < 0.05) and 50.9% (P < 0.05), respectively, reduced in the ovaries of pups from the Dx-treated mothers, when compared with the control values. There were 53.4% (P < 0.05) fewer healthy primary and 41.8% (P < 0.05) fewer healthy secondary follicles as well. The numbers of atretic primary and secondary follicles were reduced by 60.0% (P < 0.05) and 61.7% (P < 0.05), respectively. It can be concluded that fetal exposure to glucocorticoids decreased the pool of non-growing follicles in the neonatal ovary, whereas the processes of folliculogenesis and atresia remained unaffected.

Multiple treatments with SRIH-14 or octreotide affect adrenal zona glomerulosa in adult male rats.

Somatostatin analogues are currently used to treat various disorders such as hypersecretion and different neuroendocrine tumors. In this study we examined the effects on the adrenal cortex of somatostatin (SRIH-14) and octreotide administered subcutaneously twice daily for 5 days to adult male rats. Control rats received saline under the same regime. After sacrifice, the adrenal glands were removed and examined morphometrically using the M(42) multipurpose test system. Blood samples were prepared for biochemical tests. Both SRIH-14 and octreotide induced morphofunctional changes in adrenal zona glomerulosa. We found significant decreases (p < 0.05) in the absolute cell and nuclear volumes of zona glomerulosa in both experimental groups in comparison to the control. The serum aldosterone level was 11% lower (p < 0.05) in the SRIH-14 and 13% (p < 0.05) lower in the octreotide-treated group in comparison with the control group. Morphometric parameters of zona fasciculata and zona reticulata and corticosterone levels were not altered significantly (p > 0.05) in either treated group. It may therefore be concluded that both SRIH-14 and octreotide affected zona glomerulosa in the same manner by decreasing morphofunctional characteristics.

The effects of chronic SRIH-14 and octreotide administration on the pituitary-adrenal axis in adult male rats.

The effects of chronic treatments with SRIH-14 and octreotide on pituitary corticotropes (ACTH cells) and on the adrenal cortex of male Wistar rats were examined. Adult males received two daily s.c. injections of 20 microg/100 g of body weight of either SRIH-14 or octreotide for 28 consecutive days. ACTH cells were studied using a peroxidase-antiperoxidase immunocytochemical procedure. Morpho-metry was used to evaluate the changes in cell and nuclear volumes (microm3) and volume densities (%) of ACTH-immunoreactive cells. The adrenal cortex was analyzed by histological and morphometric methods. A significant (p<0.05) decrease in body weight and in the absolute weights of the pituitary and adrenal glands was observed in both treated groups. Morphometric parameters of ACTH cells in both treated groups were not significantly (p>0.05) different than in control rats. The absolute volumes of the adrenal gland and adrenal cortex were significantly (p<0.05) decreased in both treated groups. The absolute and relative volumes of the zona glomerulosa (ZG), as well as the cellular and nuclear volumes of the ZG were significantly (p<0.05) decreased in the both treated groups. In rats treated with SRIH-14 and octreotide, the absolute and relative volumes of the zona fasciculata (ZF) and zona reticularis (ZR), as well as their stereological parameters, did not change significantly (p>0.05). The aldosterone levels in the SRIH-14 and ocreotide-treated groups were significantly (p<0.05) decreased - by 13% and 19%, respectively. The concentration of ACTH and corticosterone did not change significantly. Together, these findings show that SRIH-14 and octreotide administration affected the morphological characteristics of the adrenal ZG in a similar manner, and brought about a decrease in plasma aldosterone concentration. These treatments did not affect pituitary ACTH cells or adrenal ZF and ZR functioning.

Influence of maternal dexamethasone treatment on morphometric characteristics of pituitary GH cells and body weight in near-term rat fetuses.

Growth hormone (GH) and glucocorticoids have a powerful influence on controlling fetal growth, differentiation and maturation of numerous tissues. In the present study, the effect of maternal dexamethasone (Dx) treatment on GH cells and body weight in 19- and 21-day-old rat fetuses was investigated using immunocytochemical and morphometric methods. Pregnant female rats received daily injections of 1.0-0.5-0.5 mg Dx/kg b.w. on days 16-18 of pregnancy (experimental group), while the control group received an equal volume of saline. Dx treatment of pregnant rats enhanced immunostaining intensity and significantly increased (p<0.05) GH nuclear and cell volume, as well as volume density and number of GH cells per square millimeter in 19-day-old fetuses compared to the controls. In 21-day-old fetuses after maternal Dx administration, immunoreactivity, volume density and number of GH cells remained significantly increased (p<0.05). Dx treatment of pregnant rats resulted in marked body weight reduction of 21-day-old but not 19 days old fetuses in comparison with the corresponding controls. The presented results demonstrate that maternal Dx application has pronounced effect on morphometric parameters of GH cells of 19- and 21-day-old fetuses. Also, in near-term rat fetuses body weight was largely independent of pituitary GH cell activity.

Multiple dexamethasone treatment affects morphometric parameters of gonadotrophic cells in adult female rats.

Exposure to glucocorticoids leads to numerous changes in various biological systems including the reproductive system. The aim of the present work was to find out whether dexamethasone (Dx) treatment of adult female rats would influence the histological and morphometric characteristics of the pituitary gonadotrophic cells (luteinizing--LH cells and follicle stimulating--FSH cells). One group of female Wistar rats received Dx injections on three consecutive days in doses 1.0, 0.5 and 0.5 mg/kg b.w. respectively, while the control rats were treated with equivalent volumes of saline. Experimental and control animals were sacrificed 24 h and 72 h after the last injection. The peroxidase-antiperoxidase (PAP) immunocytochemical procedure was used to study the LH and FSH cells. The stereological and morphometric analyses showed that multiple Dx treatments of female rats significantly decreased the volume of LH cells and the volume of their nuclei 24 h and 72 h after the last Dx injection in comparison with control values. At 24 h after Dx treatment, the volume density of LH cells was significantly increased, but at 72 h differences between the experimental and control groups were insignificant. The increase in number of LH cells per unit area (mm2) was significant at both timepoints (24 h and 72 h). Stereologic and morphometric characteristics of FSH cells was changed after Dx treatment in the same manner as that of LH cells, except for the volume density, where a significant increase was established 24 h and 72 h after the last Dx application. These results clearly demonstrate that 24 h and 72 h after the last of three Dx injections there were changes in the immunocytochemical and morphometric features of gonadotrophic cells.

Effects of ovariectomy and chronic estradiol administration on pituitary-thyroid axis in adult rats.

The effects of ovariectomy (Ovx) and of ovariectomy followed by chronic estradiol dipropionate administration (Ovx EDP) on the structure and function of the pituitary-thyroid axis were examined in the rat. Pituitary TSH cells and thyroid tissue were histologically, immunohistochemically and stereologically investigated. Serum TSH and T(4) levels were determined by RIA. Ovx did not affect pituitary weight, but subsequent treatment with EDP led to its more than two-fold increase (p<0.05). After ovariectomy, the cellular volume of pituitary TSH-immunoreactive cells increased by 28%, p<0.05 compared to sham-operated animals (SO). Treatment of Ovx rats with EDP partially reversed this change. However, the relative volume density of thyrotrophs decreased in comparison to the Ovx and SO groups (by 18% and 23%, p<0.05, respectively). No statistically significant differences in serum TSH levels were observed between the experimental groups. In thyroid tissue both peripheral and central follicles responded to Ovx and EDP treatments. Compared to SO rats, the relative volume densities of the follicles and colloid were increased (by 14% and 30%, p<0.05, respectively) in Ovx rats. Chronic EDP treatment of Ovx rats reversed these changes to the pre-ovariectomy state. Hyperplasia of thyroid follicular cells and a significant reduction (by 21%, p<0.05) of the serum level of T(4) were detected. In conclusion, estradiol deficiency and chronic treatment affected pituitary TSH cells and thyroid tissue. The sum effect of Ovx on the pituitary-thyroid axis was slightly stimulatory. Subsequent EDP treatment decreased thyroid functioning but at the same time preserved serum TSH at the control level.

Chronic exposure to constant light affects morphology and secretion of adrenal zona fasciculata cells in female rats.

The effect of chronic exposure to light of adult Wistar rats on growth and function of adrenal zona glomerulosa (ZG) and zona fasciculata (ZF) were examined. The females were exposed to continuous light of 600 lux for 95 days, starting on day 30 of age. The controls were kept under a 12:12 h light-dark cycle, at ambient temperature. The rats were sacrificed by decapitation and the left adrenal gland of each animal was dissected out and prepared for morphometric analyses. In animals exposed to chronic lighting, the absolute and relative volume of ZG were insignificantly increased by 5% (p>0.05) compared to controls. The volume of ZG cells and their nuclei were insignificantly changed by 1% (p>0.05) in comparison with corresponding controls. The absolute and relative volume of ZF were significantly increased (by 14 and 9%, respectively; p<0.05), as compared to controls. The volume of ZF cells and their nuclei were significantly increased (by 12 and 9%, respectively; p<0.05). Serum concentration of corticosterone was also significantly (p<0.05) increased by 13% in light-exposed group in comparison with control rats. These findings suggest that continuous exposure of female rats to constant light increased growth and secretory activity of ZF cells.

The effects of synthetic salmon calcitonin on thyroid C and follicular cells in adult female rats.

Structural and morphometric features of thyroid C and follicular cells were studied in adult rat females after treatment with synthetic salmon calcitonin (CT). The animals were chronically treated with either a low (10 IU/kg b.w) or a high (100 IU/kg b.w) dose of CT. A stereological method was applied to determine the volume density and the number of immunoreactive C cells. The height and volume density of follicular epithelium, colloid, interstitium and the follicles (epithelium plus colloid), as well as the index of activation rate were calculated. A significant decrease in body weight, as well as the volume density of immunoreactive C cells and the number of C cells per mm2, was observed in rats treated with both doses of CT. The height and volume density of follicular epithelium and follicles, as well as the index of activation rate were significantly increased in the animals given the high CT dose, while the volume densities of colloid and interstitium were reduced. No significant changes in the examined morphometric parameters were detected after treatment with the low CT dose. According to these results it can be concluded that the structural features of thyroid C and follicular cells were affected by the high dose CT treatment in the opposite manner, while the low dose CT treatment influenced only C cells.

The effect of chronic calcium treatment on thyroid C cells in ovariectomized rats.

The purpose of this study was to investigate the influence of chronic calcium treatment on the structure and function of thyroid C cells in ovariectomzed adult female rats. Eighteen 3-month-old, female Wistar rats were divided into three groups. The first group was used as the sham-operated control, and the other two were surgically ovariectomized (Ovx). One month after gonadectomy, one group of Ovx rats was injected with 28.55 mg Ca-glucoheptonate (Ca)/kg b.w., while the other two groups were chronically treated with vehicle alone (Ovx and sham control). Two months after surgery, the animals were killed. In the thyroid C cells, calcitonin (CT) was localized with the peroxidase-antiperoxidase method. Stereology was used to evaluate morphometric changes in the volume of C cells, their nuclei and relative volume density. The number of C cells per unit area was calculated. Serum CT content was determined by radioimmunoassay. After chronic Ca treatment C cells were numerous with darker cytoplasm than in C cells of sham-operated control animals, but more degranulated than the corresponding cells of Ovx rats. Their volume was significantly decreased by 14% (p < 0.05), while the number was increased by 47% (p < 0.05) in comparison with corresponding controls. Serum CT concentration was decreased by 27% (n.s.) in comparison to sham-operated control. Calcium treatment of Ovx rats led to a 32% increase of serum CT concentration in relation to untreated Ovx animals. These results suggest that chronic Ca treatment of Ovx female rats positively affected CT release from thyroid C cells, without any significant changes in morphometric parameters.

Chronic somatostatin treatment affects pituitary gonadotrophs, ovaries and onset of puberty in rats.

The effects of chronic somatostatin (SRIH-14) treatment on the pituitary gonadotrophs (FSH and LH cells) and ovaries of female Wistar rats were examined. Females were given 20 microg/100 g b.w. twice per day from the immature (23rd day) till the adult period of life (71st day). The onset of puberty was determined by daily examination for vaginal opening. The peroxidase-antiperoxidase immunocytochemical procedure was used to study the gonadotrophs. Changes in the number per unit area (mm2), cell volume and volume densities of LH- and FSH-immunoreactive cells were evaluated by morphometry and stereology. Ovaries were analysed by simple point counting of follicles and corpora lutea (CL). Follicles were divided by size according to the classification of Gaytán and Osman. The mitotic indexes of granulosa and theca cells in the follicles were estimated at all stages of folliculogenesis. The number, volume and the volume density of FSH- and LH-immunoreactive cells decreased after chronic SRIH-14 treatment, particularly the latter. In the ovary, SRIH-14 treatment decreased the number of healthy follicles at all phases of folliculogenesis, lowered the mitotic indexes of granulosa and theca cells but increased the number of atretic follicles. Healthy CL were fewer in number, while regressive CL were increased. Vaginal opening occurred at a later age in treated females. It can be concluded that chronic SRIH-14 treatment markedly inhibited LH cells and to a lesser extent FSH cells. In the ovary SRIH-14 inhibited folliculogenesis, enhanced atretic processes and lowered proliferative activity of granulosa and theca cells. It also delayed puberty onset.

The thyroid C cells of ovariectomized rats treated with estradiol.

The structure and function of thyroid C cells were studied in ovariectomized (Ovx) adult female rats without and after chronic treatment with estradiol dipropionate (EDP). A peroxidase-antiperoxidase method was applied for localization of calcitonin (CT) in the C cells. Morphometric changes in their volume, nuclei, and relative volume density were evaluated in comparison with sham-operated control rats using a stereological method. The number of C cells was calculated. CT content in the sera was determined by radioimmunoassay. Ovariectomy (Ovx) led to a 21% increase in body weight ( P<0.005), while treatment of Ovx rats with EDP decreased body weight by 25% ( P<0.01). The immunoreactivity for CT in C cells of the Ovx rats was markedly increased. Significant decreases in the volume of C cells (by 13%; P<0.05) and serum CT (by 45%) were recorded, while the C cell number increased by 59% ( P<0.05) in relation to the corresponding controls. The treatment of Ovx rats with EDP caused conspicuous degranulation of the C cells. The cellular volume was increased by 11% and serum CT by 36% in comparison with Ovx animals. At the same time a decrease in C cell number by 29% ( P<0.05) was evident. It may be concluded that estradiol deficiency after Ovx reduced the synthesis and release of CT, while chronic treatment of these animals with EDP had a positive effect on the secretory activity of thyroid C cells.

Effects of multiple somatostatin treatment on rat gonadotrophic cells and ovaries.

The effect of multiple somatostatin (SRIH-14) treatment on the pituitary gonadotrophs, follicle stimulating harmone (FSH) and luteinizing harmone (LH), and ovaries of adult female Wistar rats was examined. Females received two 20 microg/100 g body wt. doses daily subcutaneously, for five consecutive days. FSH and LH cells were studied using a peroxidase-antiperoxidase immunocytochemical procedure. Morphometry and stereology were used to evaluate changes in the number per unit area (mm2), cell volume and volume densities of LH- and FSH-immunoreactive cells. Ovaries were analysed by simple point counting of follicles and corpora lutea. Follicles were divided by size according to the classification of Gaytán and Osman. Morphometric and stereological analysis of the pituitary showed that the number, volume and the volume density of FSH- and LH-immunoreactive cells were decreased after multiple SRIH-14 treatment, particularly in the latter. In the ovary, SRIH-14 induced decreases in the number of healthy follicles in all phases of folliculogenesis and corpora lutea, but the large antral follicle stage was most affected. The number of atretic follicles was increased. It can be concluded that multiple SRIH-14 treatment markedly inhibited LH cells, but affected FSH cells as well. In the ovary, SRIH- 14 acted by inhibiting folliculogenesis and enhancing atretic processes.

Effect of centrally administered somatostatin on pituitary thyrotropes in male rats.

The effects of intracerebroventricularly administered somatostatin (SRIH-14 or -28) on growth and function of pituitary thyrotropes (TSH-cells) were examined in adult male Wistar rats. The animals were implanted with an intracerebroventricular cannula and after a recovery period, administered three 1 microg doses of SRIH-14 or -28 dissolved in 5 microl saline every second day. Controls were treated in the same way with the same volume of saline only. TSH-producing cells were studied using the peroxidase-antiperoxidase immunohistochemical procedure. Blood samples were collected for hormone (TSH) analyses 5 days after the last injection. Both SRIH-treatments significantly decreased (p < 0.05) all morphometric parameters obtained for TSH-cells in comparison with controls. The volume of TSH-cells decreased by 27%, nuclei by 44% and volume density by 33% in animals treated with SRIH-14. In animals treated with SRIH-28, these parameters were also significantly decreased (p < 0.05) (22%, 31%, and 25% respectively) compared to control rats. Serum concentrations of TSH were significantly decreased (p < 0.05) by 15% in SRIH-14- and by 12% in SRIH-28-treated animals in comparison with the controls. These observations suggest that centrally administered SRIH- 14 or -28 is specifically involved in the control of growth and secretory activity of TSH cells.

Effects of estradiol and calcium on gonadotrophic cells in middle-aged female rats.

The effects of multiple treatment with estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) or a combination of the two on gonadotrophic cells in the pituitary pars distalis of middle-aged female rats were examined. The animals were treated daily for two weeks with EDP (0.625 mg i.p./kg body weight) or Ca (11.4 mg/kg body weight) or EDP + Ca. Luteinising (LH) and follicle stimulating hormone (FSH)-producing cells were examined by immunohistochemistry using antisera to the specific (beta) beta-subunits of LH and FSH and a peroxidase-anti-peroxidase immunohistochemical procedure. Plasma levels of FSH and LH were measured by radio-immune assay. A stereological method for determining morphometric parameters in immunopositive FSH and LH cells was used. The number of gonadotrophs per unit area (mm2), their cellular volume and relative volume densities, as well as plasma levels of FSH and LH, were decreased in all treated females in comparison with the controls. The most significant decrease of these parameters was observed in EDP-treated animals. Such changes were also expressed in Ca-treated animals, but the alterations were less distinct. These results demonstrate that multiple EDP or Ca application to middle-aged female rats is able to inhibit, directly or indirectly, the morphofunctional state of gonadotrophic cells in the pituitary pars distalis.

[Hyperprolactinemia and disorders of the menstrual cycle]. / Hiperprolaktinemija i poremecaji menstrualnog ciklusa.

INTRODUCTION: The authors presented physiological conditions associated with increased prolactin values (sleep, stress, hypoglycemia, nipples stimulation, pregnancy and lactation) as well as the causes of pathological hyperprolactinemia. MATERIAL AND METHODS: The results of radioimmunoassay study have been analyzed in concern to the values of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL) obtained from the serum of 100 women with increased PRL levels. The "HOECHT" sets were used for the analysis, whereas the values up to 20.9 microgrammes per litre were rated normal. The patients were distributed into 4 groups in concern to the level of PRL increase: 1st group--twenty (n = 20) patients with PRL values from 21-29.9 micrograms/l, 90 blood samples analyzed; 2nd group-forty (n = 40) patients with PRL values f; 30-49.9 micrograms/l, 183 blood samples analyzed; 3rd group--twenty (n = 20) patients with PRL values from 50-99.9 micrograms/l, 83 blood samples analyzed; 4th group--twenty patients (n = 20) with PRL values more than 100 micrograms/l, 78 blood samples analyzed. The values of FSH and LH recorded in the women with hyperprolactinemia were compared with mean values of the same hormones presented in IU/l from the follicular phase of the cycle in the control group which comprised 50 women of reproductive age having normal ovulatory menstrual cycle. RESULTS: The mean values of FSH and LH in the 1st group have not presented with statistically significant difference in relation to the control group. Prevalence of menstrual disorders was 30%, which was statistically significantly higher than in general population. FSH values in the 2nd group were almost the same as in the control group whereas the values of LH were significantly higher. The rate of polycistic ovary syndrome (PCOS) in this group has been significant, also the increased rate of anovulatory cycles from 30 to 67.5%. A mild increase of menstrual cycle rhythm disorders, from 35 to 40% has been recorded. The values of FSH and LH in the 3rd group were significantly lower than in the control group. The significance level was higher for FSH (p < 0.01) then LH (p < 0.05). There was a sudden increase of the cycle rhythm disorders in this group reaching 90%. The 4th group presented with significantly lower values of FSH and LH in relation to the control group, whereas the cycle rhythm disorders occurred in all patients. DISCUSSION: The obtained results were compared with the literature data and some explanations given. CONCLUSIONS: The values of FSH an LH were statistically significantly lower in the 3rd and 4th group. The 2nd group was characteristic for the sudden increase of the number of anovulatory cycles from 30 to 67.5%, whereas the 3rd group presented with the abrupt increase of menstrual cycle rhythm disorders, from 40 to 90%.

[Prolactin and the menstrual cycle]. / Prolaktin i menstrualni ciklus.

In order to estimate serum prolactin levels during particular menstrual cycle phases, the authors analyzed values of follicle-stimulating hormone, luteinizing hormone and prolactin in 50 women of reproductive age with normal menstrual cycles and established ovulation. Blood samples were taken 3-5 times during follicullar, ovulatory and lutheal phase of menstrual cycle. There were 582 radioimmunoassays performed (194 per each hormone), and the upper referent value for prolactin was 20.9 ug l. The 2nd group comprised women (250) with menstrual cycle disorders (olygomenorrhea and amenorrhea) in whom involvement of hyperprolactinemia in these conditions have been done. The obtained results showed that the mean prolactin value changed in each phase of the menstrual cycle. The highest one was recorded in the periovulatory period, whereas the difference was statistically significant in relation to the follicular and lutheal phase (p < 0.05). The difference between the follicular and lutheal phase was not statistically significant, 54 (21.6%) women from the group of menstrual cycle disorders presented with increased values of prolactin.