Immunohistochemical evidence for the presence of a Vasoactive Intestinal Peptide, Neuropeptide Y, and Substance P, in rat adrenal cortex after acute heat stress

Immunohistochemistry revealed the presence of Vasoactive Intestinal Peptide (VIP), Neuropeptide Y (NPY), and the absence of Substance P (SP) immunoreactivity in the rat adrenal cortex. VIP- and NPY-immunoreactivity were detected in nerve fibers around the small blood vessels projecting into the capsule and cortical zones surrounding blood vessels and cortical cells. After acute heat stress, VIP- and NPY-immunoreactivities in the nerve fibers were reduced, probably as a result of the release of these peptides

Morfometrijsko ispitivanje nabora crevne sluzokože šarana hranjenog različitom dodatnom hranom u poluintenzivnom sistemu

U poluintenzivnom sistemu gajenja šarana u poslednjoj deceniji dolazi do promena u vrsti dodatne hrane koje povećavaju održivost proizvodnje. Sa žitarica koje su se godinama koristile kao dodata hrana, sada se prelazi na peletiranu i ekstrudiranu hranu. Ova zamena uzrokuje i promene u fiziologiji varenja riba, a kao posledica se dešavaju morfološke promene na crevima. Najčešće se na organima za varenje riba gajenih u akvakulturi ne nalaze teže histopatološke promene, pa se pribegava kvantifikaciji i morfometriji histoloških parametara koje mogu da ukažu na odstupanja od normalne građe/funkcije. Metod evaluacije dužine crevnih nabora je korišćen u velikom broju studija koje su se bavile efektima različitih tipova hrane i/ili aditiva na crevo riba. Cilj rada je da se utvrdi efekat različitih tipova hrane (žitarice, peletirana, ekstrudirana hrana) na crevo šarana merenjem dužine crevnih nabora kod riba koje su gajene poluintenzivno. Dužina crevnih nabora je korelisana sa prirastom ribe. Kretanje dužine crevnih nabora po jezerima/vrsti dodate hrane je imalo pravilan tok (Fig. 1). Najkraći crevni nabori su izmereni kod riba hranjenih žitaricama. U sva tri jezera primetan je pad dužine nabora u junu. U ovom mesecu su vrednosti najmanje, manje čak i od inicijalnih na početku eksperimenta (u aprilu), kao i vrednosti dobijenih u maju. Od juna dužina nabora creva konstantno raste kod riba u svim jezerima, bez obzira na tip dodate hrane. Rezultati su pokazali visok stepen statistički značajne korelacije između mase tela ribe i dužine crevnih nabora u svim jezerima (Tab. 2). Ukoliko se posmatraju pojedinačna jezera, vrednosti se kreću između 0.82 kod riba hranjenih peletiranom hranom, preko 0.83 kod riba hranjenih žitaricama do 0.86 kod riba hranjenih ekstrudiranom hranom. Dužina crevnih nabora je koristan parametar u eksperimentima ishrane riba i često se primenjuje u proceni efekta određene hrane na crevo riba. Smanjenje dužine nabora može da ukaže na pojavu enteritisa, na nedovoljnu ishranu ili gladovanje. Naši rezultati pokazuju pozitivnu korelaciju između mase tela ribe i dužine crevnih nabora. Oni su u skladu i sa sličnim istraživanjima koja su sprovedena na drugim životinjskim vrstama, mahom na domaćim životinjama. Crevni nabori na početku eksperimenta ne rastu, čak pokazuju trend pada sve do juna meseca i to iako ribe pokazuju konstantan prirast u svim jezerima, što se objašnjava količinom prirodne hrane koja je u to doba najveća u jezerima. Naime, na osnovu analize prirodne hrane tokom ovog perioda možemo videti da postoji velika biomasa zooplanktona u toku aprila, maja i juna dok je zoobentos na niskom nivou u svim jezerima u toku celog eksperimenta i ne prelazi 10 g/m2. Ovo uzrokuje promene u načinu ishrane ribe od juna meseca. Do tada se šaran hranio uglavnom zooplanktonom, koga je bilo u izobilju, a nakon toga, početkom juna, zbog nedostatka prirodne hrane,riba počinje da sve više konzumira dodatu, što uzrokuje i povećanje dužine crevnih nabora, s obzirom da su pelete, kojom se ribe hrane voluminoznije i većih dimenzija od zooplanktona. Hranjenjem, ribe pune creva i samim tim ih i šire, što omogućava povećanje apsorpcione površine povećanjem dužine crevnih nabora. Ovaj fenomen u stvari predstavlja adaptaciju creva na različite tipove hrane kod riba. Naše istraživanje je pokazalo da praćenje histologije šarana može da ukaže kako se odvija proces varenja zavisno od dela sezone/razvoja zajednice zooplanktona, ali i vrste dodate hrane u poluintenzivnom sistemu

Neuronal Nitric Oxide Synthase Mediates the Effect of Ethanol on IgA

Aims: We showed previously that the acute effect of ethanol on intestinal immunoglobulin A (IgA) expression might be mediated by endogenous nitric oxide (NO). To extend these findings, this study was designed to investigate a possible role of neuronal NO synthase (nNOS) in the observed phenomenon, using 7-nitroindazole (7-NI), a selective inhibitor of its activity. Methods: Adult male Wistar rats were treated with: (a) ethanol (4 g/kg, intraperitoneally, i.p.), (b) 7-NI (25 mg/kg, i.p.) followed by ethanol (4 g/kg, i.p.) 30 min later and (c) 7-NI (25 mg/kg, i.p.) followed by saline 30 min later. Untreated rats were used as controls. The concentrations of serum and intestinal IgA were measured by enzyme-linked immunosorbent assay, while the expression of nNOS was determined using western blot and immunohistochemistry. Results: Acute ethanol treatment significantly increased the concentration of IgA in the ileal extracts, whereas it decreased its serum level. Inhibition of nNOS activity by 7-NI abolished this action of alcohol on IgA. Additionally, western blot analysis revealed that the acute alcohol administration induced an increase in the expression of intestinal nNOS. Furthermore, nNOS-immunoreactive cells, observed within the lamina propria of small intestine, were numerous in ethanol-treated rats. Conclusion: Taken together, these results extended our previous findings suggesting that nNOS mediates the acute effect of ethanol on IgA and supported an immunomodulatory role of this enzyme isoform

Effects of methimazole-induced hypothyroidism on immunohistochemical, stereomorphometric and some ultrastructural characteristics of pancreatic β-cells

The function of pancreatic β-cells is to produce and secrete insulin, a crucial hormone in carbohydrate metabolism. The transcription factor PDX1 is required for insulin gene transcription and mature β-cell function. Since this factor is regulated by triiodothyronine, a disturbance in insulin biosynthesis and/or secretion is usually related to a deficiency of this hormone. In the present study, we used methods of immunohistochemistry, stereology and electron microscopy to explore the correlation between altered thyroid status and insulin synthesis/secretion in a model of methimazole-induced hypothyroidism in rats. In hypothyroid animals fewer functional PDX1-positive β-cells were detected in the islets of Langerhans, while insulin immunostaining was stronger. Stereological analysis of β-cell granules revealed more numerous immature insulin granules in hypothyroid rats. Taken together, these data suggest that the applied treatment caused impaired insulin synthesis and secretion. Rare cells with granules characteristic for both α- and β-cells observed in hypothyroid animals could provide functional compensation for diminished insulin synthesis

Calcium-SANDOZ®-induced erythrocyte exovesiculation and internalization of hemichromic material into rat brown adipocytes

An ultramicroscopic study of brown adipose tissue (BAT) of rats treated with Ca-SANDOZ® (480 mg/l) for 3 days, revealed erythrocyte exovesiculation and migratory erythrocytic complexes from the capillaries to adipocyte cytoplasm and mitochondria. Two types of erythrocytic material transfer were observed: (i) numerous exocytic vesicles with electron dense material leaving the erythrocytes; (ii) furcated complexes with microholes, embedded in amorphous material. The content of red blood cell (RBC) complexes passed through the capillaries and transferred to the brown adipocytes where it was detectable in the cytoplasm and mitochondria. Light microscopy confirmed sphenoechinocytic transformation of the RBCs in the blood smears of the Ca-SANDOZ® treated rats

BRCA1 and TOP2A gene amplification and protein expression in four molecular subtypes of breast cancer

We studied TOP2A amplification (using FISH methods), and TOP2A and BRCA1 protein overexpression (immunohistochemistry) in four molecular subtypes of breast cancer. Of 53 patients, 32 showed TOP2A and 38 showed BRCA1 overexpression. The highest percentage of TOP2A amplification (47.8%) and deletion (13%) was detected in Luminal B subtypes. Of 11 Luminal B tumors with TOP2A amplification, 9 (81.8%) overexpressed TOP2A. BRCA1 protein overexpression showed significant positive correlation with TOP2A protein expression. BRCA1 and TOP2A proteins exhibited similar patterns of expression in Luminal B and triple-negative breast cancer, suggesting the same prognosis in those patients

Blokada sinteze azot-monoksida stimuliše aktivnost korteksa nadbubrežne žlezde

Objective. Although it is known that nitric oxide modulates the activity of hypothalamic-pituitary-adrenal axis, its functional significance has not been elucidated. Additionally, there is no information on the effect of nitric oxide on cortical expression of glucocorticoid receptor. The purpose of this study was to investigate the influence of endogenous nitric oxide on structure and function of rat adrenal cortex and adrenocortical expression of glucocorticoid receptor. Methods. Adult female Wistar rats showing diestrus day 1 were treated with Nω-nitro-L-arginine-methyl ester (LNAME), an inhibitor of all three isoforms of nitric oxide synthase, at a dose of 30 mg/kg, subcutaneously. The concentrations of adrenocorticotropic hormone (ACTH) and corticosterone were determined by radioimmunoassay. Stereological and immunohistochemical analyses were performed on paraffin sections. Results. Blockade of nitric oxide production significantly increased blood levels of ACTH and corticosterone. Stereological analysis showed that treatment with L-NAME significantly decreased numerical density of the cells in all cortical zones. Consistent with the decreased numerical density, L-NAME significantly increased the volume of cells in cortical zones. Inhibition of nitric oxide synthesis decreased expression of glucocorticoid receptor in zona fasciculata and zona reticularis. Conclusion. Obtained results indicate that endogenous nitric oxide inhibits activity of adrenal cortex and modulates expression of glucocorticoid receptor.Cilj. Iako je poznato da azot-monoksid moduliše aktivnost hipotalamo-hipofizno-nadbubrežne osovine, funkcionalni značaj tog delovanja nije rasvetljen. Pored toga, dejstvo azot-monoksida na kortikalnu ekspresiju glukokortikoidnog receptora nije istraženo. Cilj ovog rada bio je da se ispita uticaj endogenog azot-monoksida na strukturu i funkciju kore nadbubrežne žlezde pacova i adrenokortikalnu ekspresiju glukokortikoidnog receptora. Metode. Odrasle ženke pacova Wistar soja u metestrusnoj fazi ciklusa tretirane su Nω-nitro-L-arginin metil esterom (L-NAME), inhibitorom sve tri izoforme azotmonoksid sintaze, u dozi 30 mg/kg, subkutano. Koncentracije adrenokortikotropnog hormona (ACTH) i kortikosterona u krvi određene su radioimunološkom metodom. Na parafinskim presecima nadbubrežne žlezde izvršena je stereološka i imunohistohemijska analiza korteksa. Rezultati. Blokada sinteze azot-monoksida značajno povećava koncentracije ACTH i kortikosterona u krvi. Stereološka analiza je pokazala da tretman sa L-NAME značajno smanjuje numeričku gustinu ćelija u svim kortikalnim zonama. U skladu sa smanjenom numeričkom gustinom, L-NAME značajno povećava volumen ćelija u sve tri zone korteksa. Inhibicija sinteze azot-monoksida smanjuje ekspresiju glukokortikoidnog receptora u zoni fascikulati i zoni retikularis. Zaključak. Dobijeni rezultati ukazuju na mogućnost da endogeni azot-monoksid inhibiše aktivnost kore nadbubrežne žlezde i da moduliše ekspresiju glukokortikoidnog receptora

Chronic psychological stress activates BMP4-dependent extramedullary erythropoiesis

Psychological stress affects different physiological processes including haematopoiesis. However, erythropoietic effects of chronic psychological stress remain largely unknown. The adult spleen contains a distinct microenvironment favourable for rapid expansion of erythroid progenitors in response to stressful stimuli, and emerging evidence suggests that inappropriate activation of stress erythropoiesis may predispose to leukaemic transformation. We used a mouse model to study the influence of chronic psychological stress on erythropoiesis in the spleen and to investigate potential mediators of observed effects. Adult mice were subjected to 2hrs daily restraint stress for 7 or 14 consecutive days. Our results showed that chronic exposure to restraint stress decreased the concentration of haemoglobin in the blood, elevated circulating levels of erythropoietin and corticosterone, and resulted in markedly increased number of erythroid progenitors and precursors in the spleen. Western blot analysis revealed significantly decreased expression of both erythropoietin receptor and glucocorticoid receptor in the spleen of restrained mice. Furthermore, chronic stress enhanced the expression of stem cell factor receptor in the red pulp. Moreover, chronically stressed animals exhibited significantly increased expression of bone morphogenetic protein 4 (BMP4) in the red pulp as well as substantially enhanced mRNA expression levels of its receptors in the spleen. These findings demonstrate for the first time that chronic psychological stress activates BMP4-dependent extramedullary erythropoiesis and leads to the prolonged activation of stress erythropoiesis pathways. Prolonged activation of these pathways along with an excessive production of immature erythroid cells may predispose chronically stressed subjects to a higher risk of leukaemic transformation

The Acute Effect of Ethanol on Adrenal Cortex in Female Rats-Possible Role of Nitric Oxide

Aims: The present study was designed to investigate a possible role of endogenous nitric oxide (NO) in the adrenal response to an acute alcohol administration in female rats. To this end, N(omega)-nitro-l-arginine-methyl ester (l-NAME), a competitive inhibitor of all isoforms of NO synthase, was used. Methods: Adult female Wistar rats showing diestrus Day 1 were treated with: (a) ethanol (2 or 4 g/kg, intraperitoneally); (b) l-NAME (30 or 50 mg/kg, subcutaneously) followed by either ethanol or saline 3 h later. Untreated and saline-injected rats were used as controls. The animals were killed 30 min after last injection. Adrenal cortex was analyzed morphometrically, and plasma levels of adrenocorticotropic hormone (ACTH) and serum concentrations of corticosterone were determined. Results: Acute ethanol treatment enhanced the levels of ACTH and corticosterone in a dose-dependent manner. Stereological analysis revealed that acute alcohol administration induced a significant increase in absolute volume of the cortex and the zona fasciculata (ZF). In addition, ethanol at a dose of 4 g/kg increased volume density and length of the capillaries in the ZF. However, other stereological parameters were unaffected by alcohol exposure. Pretreatment with both doses of l-NAME had no effect on ethanol-induced changes. Conclusion: Obtained findings indicate that acute ethanol treatment stimulates the activity of the adrenal cortex and that this effect is not mediated by endogenous NO in female rats under these experimental conditions