Investigation of Stress Adaptation Systems of Pituitary Adenylate-cyclase Activating Polypeptide Deficient Mice in Acute and Chronic Models

János Selye defined the classic concept of stress as a „nonspecific response of the body to any threatening demand” (1936). More recently, McEwen (2000) defined stress as a real or interpreted threat to the physiological or psychological integrity on an individual that results in physiological and/or behavioral responses”. The prevalence of mood disorders in the western type of communities is continuously increasing: major depression is now the most common reason for chronic disability causing significant social and financial load on the healthcare systems (WHO, 2016). The complex neurobiological and neurochemical background of depression is not fully understood. Despite the fact that multiple groups of medicines are available for the treatment, there are patients who do not respond to the pharmacotherapy. Additionally, due to the wide range of side effects sometimes it is not possible to introduce the optimal therapeutic strategy. Therefore, to find more effective therapeutical strategies it is essential to deeper understand the pathophysiological background of the psychopathology. To reach this purpose the identification of new key mediators and drug targets is required. This may be possible if the imperative need for new well characterized and more reliable animal models will be fulfilled. Numerous groups neuropeptides and their receptors have been found to exert a tonic regulatory role in stress adaptation-response both in animals and humans. Clinical trials have been lunched to test if newly designed pharmacological agents targeted to the neuropeptide receptors may be applied as useful tools in the management of depression. (Lin 2012, Kormos and Gaszner 2013, Catena-Dell’Osso at al., 2013). The main goal of my PhD research program was to examine the stress adaptation systems in acute and chronic mouse models with the hope that our new findings may contribute to gain deeper insight into the neurobiological background of depression and may help to identify new therapeutical approaches

The anti-inflammatory effect of dimethyl trisulfide in experimental acute pancreatitis

Various organosulfur compounds, such as dimethyl trisulfide (DMTS), display anti-inflammatory properties. We aimed to examine the effects of DMTS on acute pancreatitis (AP) and its mechanism of action in both in vivo and in vitro studies. AP was induced in FVB/n mice or Wistar rats by caerulein, ethanol-palmitoleic acid, or L-ornithine-HCl. DMTS treatments were administered subcutaneously. AP severity was assessed by pancreatic histological scoring, pancreatic water content, and myeloperoxidase activity measurements. The behaviour of animals was followed. Pancreatic heat shock protein 72 (HSP72) expression, sulfide, and protein persulfidation were measured. In vitro acinar viability, intracellular Ca 2+ concentration, and reactive oxygen species production were determined. DMTS dose-dependently decreased the severity of AP. It declined the pancreatic infiltration of leukocytes and cellular damage in mice. DMTS upregulated the HSP72 expression during AP and elevated serum sulfide and low molecular weight persulfide levels. DMTS exhibited cytoprotection against hydrogen peroxide and AP-inducing agents. It has antioxidant properties and modulates physiological but not pathophysiological Ca 2+ signalling. Generally, DMTS ameliorated AP severity and protected pancreatic acinar cells. Our findings indicate that DMTS is a sulfur donor with anti-inflammatory and antioxidant effects, and organosulfur compounds require further investigation into this potentially lethal disease

Characterization of Neurons Expressing the Novel Analgesic Drug Target Somatostatin Receptor 4 in Mouse and Human Brains

Somatostatin is an important mood and pain-regulating neuropeptide, which exerts analgesic, anti-inflammatory, and antidepressant effects via its Gi protein-coupled receptor subtype 4 (SST4) without endocrine actions. SST4 is suggested to be a unique novel drug target for chronic neuropathic pain, and depression, as a common comorbidity. However, its neuronal expression and cellular mechanism are poorly understood. Therefore, our goals were (i) to elucidate the expression pattern of Sstr4/SSTR4 mRNA, (ii) to characterize neurochemically, and (iii) electrophysiologically the Sstr4/SSTR4-expressing neuronal populations in the mouse and human brains. Here, we describe SST4 expression pattern in the nuclei of the mouse nociceptive and anti-nociceptive pathways as well as in human brain regions, and provide neurochemical and electrophysiological characterization of the SST4-expressing neurons. Intense or moderate SST4 expression was demonstrated predominantly in glutamatergic neurons in the major components of the pain matrix mostly also involved in mood regulation. The SST4 agonist J-2156 significantly decreased the firing rate of layer V pyramidal neurons by augmenting the depolarization-activated, non-inactivating K+ current (M-current) leading to remarkable inhibition. These are the first translational results explaining the mechanisms of action of SST4 agonists as novel analgesic and antidepressant candidates

Vegyszeres nyomelőhívási eljárások és kábítószeranalízis a rendőrségen

Vegyszeres nyomelőhívási eljárások részletes bemutatása. Vérnyomok előhívása, kutatása. Porózus és nem porózus felületek kezelése. Kábítószeranalízis. Metalográfia.BscBiomérnökg

A tebainban gazdag, pelletált Tebona mákgubó extrakciója oldószeresen

A szakdolgozatom kísérleti témájának kiindulópontja az volt, hogy hogyan is lehetne a pelletált Tebona mákgubóból hatékonyabban, gazdaságosabban a tebain hatóanyagot kinyerni. Mivel a gyár célja a pelletált állagú mákgubóból való kiindulás, és a morfin mellet a mákban található egyéb alkaloidok hatékonyabb kioldása, ezért szükség volt egy alternatív eljárás kidolgozására, mely eljárás a kísérletem alapját szolgáltatta.MSc/MABiomérnökg

Perifériás ideg epineuralis metilénkékfestése kadáverkézen

Bevezetés: A parciális aponeurectomia a Dupuytren-kontraktúra kezelésére leggyakrabban végzett műtéti eljárás. A betegségben kialakuló patológiás szövet megváltoztathatja a digitális ideg anatómiai elhelyezkedését, ami megne- hezíti az ideg műtét közbeni lokalizációját és dissectióját, és növeli a iatrogén idegsérülés kockázatát. Intraoperatív idegfestési eljárással az ideg lokalizációja megkönnyíthető lenne, ezáltal a iatrogén idegsérülés kockázata is csökkenne. Állatkísérleteinkben korábban igazoltuk, hogy metilénkékoldattal a perifériás ideg in vivo megfesthető az ideg struktúrájának és funkciójának károsítása nélkül. Célkitűzés: A patkány nervus ischiadicus modellen már sikeresen alkalmazott metilénkékoldattal végzett idegfestési eljárás hatékonyságának vizsgálata humán kadáver digitális idegen. Módszer: Vizsgálatunk első fázisában formalinnal fixált kézen négy digitális ideg epineuralis festését végeztük el 40 μl 1 : 80-as hígítású metilénkékoldattal. A második vizsgálatban fixáción át nem esett kadáverkézen hat digitális idegfestését végeztük. A megfesthető idegszakasz hosszának növelésére két ideg festéséhez 200 μl metilénkékoldatot használtunk. Eredmények: Az epineuralis idegjelölés formalinfixált idegeken nem működött ideálisan. Friss, formalinos fixáláson át nem esett humán kadáver digitális idegen az idegfestési eljárást sikeresen alkalmaztuk, a megfestett idegszakasz tekintetében állatkísérletes eredményeinket reprodukálni tudtuk. 40 μl 1 : 80-as hígítású metilénkékoldattal átlagosan 13 mm-es, míg 200 μl oldat használatával 18 mm-es idegszakaszt sikerült megfesteni. Következtetés : Formalinnal fixált digitális ideg festése a fixáció következtében fellépő szöveti zsugorodás miatt korlátozottan lehetséges. Formalinos fixáláson át nem esett digitális idegek esetén a megtartott anatómiai viszonyok mellett 18 mm-es idegszakasz megfesthető. További vizsgálatokat tervezünk Dupuytren-szövettel és hegszövettel körülvett digitális idegen, a technika kézsebészeti műtétek során történő alkalmazhatóságának megítélésére

The behavioral phenotype of pituitary adenylate-cyclase activating polypeptide-deficient mice in anxiety and depression tests is accompanied by blunted c-Fos expression in the bed nucleus of the stria terminalis, central projecting Edinger-Westphal nucleus, ventral lateral septum, and dorsal raphe nucleus

Pituitary adenylate-cyclase activating polypeptide (PACAP) has been implicated in the (patho)physiology of stress-adaptation. PACAP deficient (PACAP(-/-)) mice show altered anxiety levels and depression-like behavior, but little is known about the underlying mechanisms in stress-related brain areas. Therefore, we aimed at investigating PACAP(-/-) mice in light-dark box, marble burying, open field, and forced swim paradigms. We also analyzed whether the forced swim test-induced c-Fos expression would be affected by PACAP deficiency in the following stress- related brain areas: magno- and parvocellular paraventricular nucleus of the hypothalamus (PVN); basolateral (BLA), medial (MeA), and central (CeA) amygdaloid nuclei; ventral (BSTv), dorsolateral (BSTdl), dorsomedial (BSTdm), and oval (BSTov) nuclei of the bed nucleus of stria terminalis; dorsal (dLS) and ventral parts (vLS) of lateral septal nucleus, central projecting Edinger-Westphal nucleus (EWcp), dorsal (dPAG) and lateral (lPAG) periaqueductal gray matter, dorsal raphe nucleus (DR). Our results revealed that PACAP(-/-) mice showed greatly reduced anxiety and increased locomotor activity compared with wildtypes. In forced swim test PACAP(-/-) mice showed increased depression-like behavior. Forced swim exposure increased c-Fos expression in all examined brain areas in wildtypes, whereas this was markedly blunted in the DR, EWcp, BSTov, BSTdl, BSTv, PVN, vLS, dPAG, and in the lPAG of PACAP(-/-) mice vs. wildtypes, strongly suggesting their involvement in the behavioral phenotype of PACAP(-/-) mice. PACAP deficiency did not influence the c-Fos response in the CeA, MeA, BSTdm, and dLS. Therefore, we propose that PACAP exerts a brain area- specific effect on stress-induced neuronal activation and it might contribute to stress-related mood disorders

Aging Changes the Efficacy of Central Urocortin 2 to Induce Weight Loss in Rats

Middle-aged obesity and aging cachexia present healthcare challenges. Central responsiveness to body-weight-reducing mediators, e.g., to leptin, changes during aging in a way, which may promote middle-aged obesity and aging cachexia. Leptin is connected to urocortin 2 (Ucn2), an anorexigenic and hypermetabolic member of the corticotropin family. We aimed to study the role of Ucn2 in middle-aged obesity and aging cachexia. The food intake, body weight and hypermetabolic responses (oxygen consumption, core temperature) of male Wistar rats (3, 6, 12 and 18 months) were tested following intracerebroventricular injections of Ucn2. Following one central injection, Ucn2-induced anorexia lasted for 9 days in the 3-month, 14 days in the 6-month and 2 days in the 18-month group. Middle-aged 12-month rats failed to show anorexia or weight loss. Weight loss was transient (4 days) in the 3-month, 14 days in the 6-month and slight but long-lasting in the 18-month rats. Ucn2-induced hypermetabolism and hyperthermia increased with aging. The age-dependent changes in the mRNA expression of Ucn2 detected by RNAscope in the paraventricular nucleus correlated with the anorexigenic responsiveness. Our results show that age-dependent changes in Ucn2 may contribute to middle-aged obesity and aging cachexia. Ucn2 shows potential in the prevention of middle-aged obesity