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The lemon flavonoid extract Eriomin® (LE), which is rich in eriocitrin, has demonstrated antioxidant and anti-inflammatory properties in both animal and human studies. Given the established interplay among aging, oxidative stress, and inflammation, this study investigated the influences of LE on the pituitary–adrenal (PA) axis in aged rats and its potential to mitigate age-related physiological changes in this system. The effects of LE (40 mg/kg/day suspended in sunflower oil) on the morphofunctional properties of the PA axis were studied in 24-month-old male Wistar rats following four weeks of oral treatment. Control groups included vehicle-treated (sunflower oil; CON) and untreated intact controls (ICON). Stereological and imaging analyses revealed no significant changes in pituitary ACTH cells; however, Pomc gene expression was significantly downregulated in the LE group compared to both controls (p ≤ 0.05). LE treatment resulted in a significant reduction in adrenal gland weight (p ≤ 0.05), adrenal gland volume (p ≤ 0.01), zona fasciculata (ZF) volume (p ≤ 0.01) and ZF cell volume (p ≤ 0.05). These changes were accompanied by a significant decrease in serum corticosterone levels (p ≤ 0.05). In conclusion, LE downregulated PA axis activity in aged rats. Considering the association between age-related increases in PA activity and adverse health outcomes, citrus flavonoid extracts such as LE may hold promise as anti-aging supplements aimed at mitigating age-related stress dysregulation.

The role of membrane fluidity in determining red blood cell (RBC) deformability has been suggested by a number of studies. The present investigation evaluated alterations of RBC membrane fluidity, deformability and stability in the presence of four linear alcohols (methanol, ethanol, propanol and butanol) using ektacytometry and electron paramagnetic resonance (EPR) spectroscopy. All alcohols had a biphasic effect on deformability such that it increased then decreased with increasing concentration; the critical concentration for reversal was an inverse function of molecular size. EPR results showed biphasic changes of near-surface fluidity (i.e., increase then decrease) and a decreased fluidity of the lipid core; rank order of effectiveness was butanol > propanol > ethanol > methanol, with a significant correlation between near-surface fluidity and deformability (r = 0.697; p<0.01). The presence of alcohol enhanced the impairment of RBC deformability caused by subjecting cells to 100 Pa shear stress for 300 s, with significant differences from control being observed at higher concentrations of all four alcohols. The level of hemolysis was dependent on molecular size and concentration, whereas echinocytic shape transformation (i.e., biconcave disc to crenated morphology) was observed only for ethanol and propanol. These results are in accordance with available data obtained on model membranes. They document the presence of mechanical links between RBC deformability and near-surface membrane fluidity, chain length-dependence of the ability of alcohols to alter RBC mechanical behavior, and the biphasic response of RBC deformability and near-surface membrane fluidity to increasing alcohol concentrations.

Soy isoflavone’s (genistein and daidzein in particular) biological significance has been thoroughly studied for decades, so we started from the premise that refreshed investigation approach in this field should consider identification of their new molecular targets. In addition to recently described epigenetic aspects of polyphenole action, the cell membrane constituents-mediated effects of soy isoflavones are worthy of special attention. Accordingly, the expanding concept of membrane steroid receptors and rapid signaling from the cell surface may include the prominent role of these steroid-like compounds. It was observed that daidzein strongly interacts with membrane estrogen receptors in adrenal medullary cells. At low doses, daidzein was found to stimulate catecholamine synthesis through extracellular signal-regulated kinase 1/2 or protein kinase A pathways, but at high doses, it inhibited catecholamine synthesis and secretion induced by acetylcholine. Keeping in mind that catecholamine excess can contribute to the cardiovascular pathologies and that catecholamine lack may lead to depression, daidzein application promises to have a wide range of therapeutic effects. On the other hand, it was shown in vitro that genistein inhibits LNCaP prostate cancer cells invasiveness by decreasing the membrane fluidity along with immobilization of the androgen receptor containing membrane lipid rafts, with down regulation of the androgen receptors and Akt signaling. These data are promising in development of the molecular pharmacotherapy pertinent to balanced soy isoflavone treatment of cardiovascular, psychiatric, and steroid-related malignant diseases.

As a consequence of global warming, the increase in the average annual temperature is observed, while the living organisms actively adapt to these changes. High environmental temperature initiates numerous physiological, autonomic, and behavioral responses, and activates the stress response. Thus, the aim of the study was to investigate effect of a moderate increase in ambient temperature on the activity of the hypothalamic-pituitary-adrenocortical (HPA) axis by determining histological changes in adrenal glands and hormonal levels in adult male rats. In this experimental study, the morpho-functional state of adrenal glands was estimated by stereological evaluation of parameters, including the adrenal volume, adrenocortical cell/nuclear size and number, and the volume density of vascular tissues after four days of exposure to a moderate increase in ambient temperature of 35 ± 1˚C. Novelli histochemical and vascular endothelial growth factor (VEGF) immunohistochemical staining provided insight into the adrenal gland vascular network. Additionally, the adrenal levels of aldosterone, corticosterone, and pituitary adrenocorticotropic hormone (ACTH) were determined as crucial indicators of the hypothalamic-pituitaryadrenocortical (HPA) axis activity. Prolonged exposure to a moderate increase in ambient temperature for four days resulted in a significant increase in ACTH level up to 24%, which altered adrenal glands both structurally and functionally. The adrenocortical volume and number of cells in all cortical zones were markedly increased (P<0.05). A statistically significant increase was shown in the level of aldosterone (16%) and corticosterone (25%) in serum levels of individuals. Increased activity of the HPA axis reflects the response to a moderate increase in ambient temperature during four days, showing the capacity of the HPA axis to adapt the organism to daily temperature changes.

Global warming causes an increased ambient temperature and prolonged heatwaves during the summer, which represent stressogenic factors affecting the hypothalamo-pituitary-adrenocortical (HPA) axis in mammals. The aim of this study was to investigate the effects of long-term (7-60 days) exposure to moderately elevated ambient temperature (35 ± 1°C) on the histological aspect and secretory ability of pituitary adrenocorticotropic (ACTH) cells, as well as on the corticosterone output, in adult rats. Stereological parameters of ACTH cells were estimated upon immunohistochemistry. The blood concentrations of ACTH and corticosterone were determined by immunoassays. The volume of ACTH cells in rats exposed to moderately high temperature for 7, 14, 21, 30 and 60 days decreased (p<0.05) by 18.1%, 14.5%, 13.5%, 8.6% and 14.2% respectively, compared to the same parameter in the controls. The volume density of ACTH cells in the groups exposed to elevated temperature for 7, 14, 21, 30 and 60 days decreased (p<0.05) by 40.0%, 33.3%, 26.7%, 13.3% and 26.7% respectively, in comparison with control rats. The plasma concentration of ACTH varied differently (p<0.05) with the duration of exposure to the elevated temperature. The serum concentration of corticosterone was decreased (p<0.05) by 54.9%, 24.4%, 29.9%, 21.1% and 24.4% in groups subjected to moderately high temperature for 7, 14, 21, 30 and 60 days respectively, all compared to the control value. Despite some signs of functional recovery of ACTH cells during the treatment, the impression is that the long-term character of this stressor overcomes the capacity of the HPA axis for resistance.

Soy isoflavones are diphenolic compounds that are frequently used for alternative treatment of ageing symptoms in both genders. They operate at principally two hierarchical levels of functional organization – cellular and molecular, while these ‘types’ of action appear to have indefinite borders. Soy isoflavone action at the cellular level involves inter alia the effects on cell mechanics. This epigenetic and modular determinant of cell function and fate is defined by: the anchorage to extracellular matrix (ECM) and neighboring cells, cytoskeleton organization, membrane tension and vesicle trafficking. Soy isoflavones have been reported to: (i) generally fashion an inert cell phenotype in some cancers and enhance the cell anchorage in connective tissues, via the effects on ECM proteins, focal adhesion kinases-mediated events and matrix metalloproteinases inhibition; (ii) affect cytoskeleton integrity, the effects being related to Ca 2+ ions fluxes and involving cell retraction or differentiation/proliferation-related variations in mechanical status; (iii) increase, remain “silent” or decrease membrane tension/fluidity, which depends on polarity and a number and arrangement of functional groups in applied isoflavone; (iv) provoke inhibitory effects on vesicle trafficking and exo-/endocytosis, which are usually followed by changed cell morphology. Here we present and discuss the abundance of effects arising from cells’ “encounter” with soy isoflavones, focusing on different morphofunctional definers of cell mechanics.

The aim of the present study was to determine and elaborate on all changes in old-aged (OA) versus young-aged (YA) rat thyroids by using stereological, ultrastructural, hormonal, and gene expression analyses. We used 4- and 24-month-old male Wistar rats in our evaluation, presenting all changes in comparison with YA rats. Results showed that the thyroid parenchyma was characterized by higher absolute volumes of the gland, colloid, epithelium, and interstitium by 135, 135, 140, and 142% (p < 0.05) respectively, while the relative volumes of colloid and glands were unchanged. Ultrastructural analysis revealed less active glands, with smaller amounts of lysosomes, thyroglobulin (Tg) granules, and microvilli in the luminal colloid. Optical density values for thyroid peroxidase (TPO), Tg, and vascular-endothelial growth factor immunostaining remained unchanged; however, TPO and Tg exhibited visually stronger expression in small active follicles. Thyroxine (T4)-Tg, the relative intensity of fluorescence (RIF), serum T4, and the sodium-iodide symporter immunohistochemical and gene expressions decreased by 20, 40, 29, and 31% (p < 0.05), respectively, in OA thyroids. Pituitary thyroid-stimulating hormone (TSH) RIF increased by 44% (p < 0.05), but the TSH serum concentration remained unchanged. In conclusion, the obtained results indicate depression of the thyroid gland synthetic and secretory capacity with advanced age.

The pathological phenomenon of somatopause, noticeable in hypogonadal ageing subjects, is based on the growth hormone (GH) production and secretion decrease along with the fall in GH binding protein and insulin-like growth factor 1 (IGF-1) levels, causing different musculoskeletal, metabolic and mental issues. From the perspective of safety and efficacy, GH treatment is considered to be highly controversial, while some other therapeutic approaches (application of IGF-1, GH secretagogues, gonadal steroids, cholinesterase-inhibitors or various combinations) exhibit more or less pronounced weaknesses in this respect. Soy isoflavones, phytochemicals that have already demonstrated the health benefits in treated elderly, at least experimentally reveal their potential for the somatopausal symptoms remediation. Namely, genistein enhanced GHRH-stimulated cAMP accumulation and GH release in rat anterior pituitary cells; refreshed and stimulated the somatotropic system (hypothalamic nuclei and pituitary GH cells) function in a rat model of the mild andropause, and stimulated the GH output in ovariectomized ewes as well as the amplitude of GH pulses in the rams. Daidzein, on the other hand, increased body mass, trabecular bone mass and decreased bone turnover in the animal model of severe andropause, while both isoflavones demonstrated blood cholesterol-lowering effect in the same model. These data, which necessarily need to be preclinically and clinically filtered, hint some cautious optimism and call for further innovative designing of balanced soy isoflavone-based therapeutics.

In areas with moderate continental climate, increased average ambient temperature during the summer represents a stressogenic factor that affects the hypothalamo-pituitaryadrenocortical axis in mammals. Therefore, we wanted to examine the effects of 4 days of constant exposure to moderately elevated ambient temperature (35 ± 1oC) on the histomorphometric and immunofl uorescent characteristics, as well as on the hormonal secretion of pituitary corticotropes (ACTH) cells in adult male rats. In comparison with the controls kept at 20 ± 2oC, a signifi cant increase (p<0.05) of the absolute and relative pituitary weight (23.1% and 36.1%, respectively) was registered after exposure to heat. The localization, as well as the shape of the ACTH cells in the heat exposed group was not signifi cantly altered, but their immunopositivity was weaker. After 4 days of heat exposure, a weaker signal confi rmed the relative fl uorescence intensity of the ACTH cells (15.3%, p<0.05). In heat exposed rats, an increase of the cellular and nuclear volumes of immunolabelled ACTH cells and decrease of their volume density (6.9%, 14.3% and 20.0%, respectively; p<0.05) was registered. Observed histomorphometric and immunofl uorescent features of the pituitary ACTH cells were in accordance with the increased (p<0.05) value of plasma adrenocorticotropic hormone (ACTH) by 23.7% compared to the control rats. It can be concluded that the 4-day exposure to moderately elevated ambient temperature intensifi es pituitary ACTH secretion in adult male rats.

Andropause, the culminating phase of male ageing, is characterized by deregulation of the hypothalamic-pituitarygonadal axis and low circulating free testosterone. The aim of this study was to investigate the immunohistomorphometric characteristics of the pituitary gonadotropic . follicle-stimulating hormone (FSH) and the luteinizing hormone (LH) producing cells after testosterone application in a rat model of the andropause. Middle-aged Wistar rats were divided into orchidectomized (ORX; n=8) and testosterone treated orchidectomized (ORX+T; n=8) groups. Testosterone propionate (5 mg/kg b.m./day) was administered for three weeks, while the ORX group received the vehicle alone. Immunohistochemically stained FSH and LH cells underwent morphometric and optical density-related analysis, while circulating concentrations of the sex steroids were measured by immunoassays. Serum concentrations of testosterone and estradiol were significantly (p<0.05) increased by 24 and 2.7 fold respectively, compared to the ORX group. The volume of FSH and LH cells was significantly (p<0.05) decreased by 51.3% and 56.6% respectively, in comparison with ORX rats. Relative volume density of FSH and LH cells was also significantly (p<0.05) decreased by 54.0% and 72.8% respectively, compared to the ORX group. Results related to the optical density of gonadotropic cells (reflecting their hormonal content) were in line with the morphometric findings . this parameter of FSH and LH cells was significantly (p<0.05) decreased by 25.7% and 16.2% respectively, in comparison with ORX rats. Conclusion: In conclusion, applied testosterone increased the serum concentrations of sex steroids, as well as it decreased morphometric parameters and optical density of gonadotropic cells in ORX rats.