Examination of the Lack of Endogenous Pituitary Adenylate Cyclaseactivating Polypeptide (PACAP) in Knockout Mice in the Auditory System, Vascular System and During Tooth Development

A Ph.D. értekezésem alapjául szolgáló kutatások során hypophysis adenilát-cikláz aktiváló polipeptid (PACAP)-génkiütött egerekben vizsgáltuk, hogy az endogén PACAP hiánya milyen módon befolyásolja különböző szervrendszerek működését. Értekezésemben ezen vizsgálatok közül a hallórendszerrel, az érrendszerrel és a fogfejlődéssel kapcsolatos legújabb eredmények kerülnek leírásra

Operációkutatási módszerek műszaki informatikai rendszerek analízisében és verifikációjában = Operation Research Methods for the Analysis and Verification of Information Technology Systems

Kidolgoztuk a Petri-hálók és produkciós hálók (PNS) egységes szemléletű leírását. Megfogalmaztuk az "optimális trajektória generálásának" problémáját Petri-hálós modellekre. A megoldásként kidolgozott és implementált algoritmus egyúttal temporális logikai követelményeket is vizsgál a modellen. Az algoritmust gyorsítottuk a PNS logikai bázisa fölötti kereséssel. A SPIN modellellenőrzőt magát használva egy másik megoldást is adtunk a problémára, valamint gráftranszformációs rendszerek optimalizálására. Megadtuk a lineáris korlátozási feltételekkel adott szeparábilis konkáv minimalizálási feladat egy elégséges optimalitási kritériumát, mely a Branch-and-Bound típusú algoritmusban használható fel megállási kritériumként. A magasszintű leírásokból a Petri-hálós modellbe történő transzformációkat matematikai alapokon definiáltuk, megvalósításukra automatikus modelltranszformációs megoldást dolgoztunk ki: egy algoritmust, amely GRM profillal adott modellből generálja a Petri-hálót, és egy általános algoritmust, amely UML modellekből származtat a diagnosztika alapjául szolgáló modelleket. Megvizsgáltuk ezen modellek illeszthetőségét a szabványokhoz. Multiprocesszoros rendszerek diagnosztizálására egy PNS technikákat használó algoritmust adtunk, melynek várható hatékonyságát igazoltuk. Munkálatok folytak a diagnosztika tesztalapú megközelítésére, és diagnosztikai modellek kísérletes paraméterezésére. Kísérleteket végeztünk az IBM Holosofx ipari workflow modellező eszköz illesztésére. | A unified treatment for Petri nets and process network (PNS) problems was defined. The 'optimal trajectory generation problem' for Petri nets was defined. Elaboration and implementation of an algorithm that is able not only to give the optimal trajectory but to verify temporal logic requirements for Petri nets. This algorithm was accelerated using Branch-and-Bound method over the logical basis of the feasible process networks. Another algorithm to solve the problem using only the SPIN model checker was elaborated. The optimization of graph transformation systems with time was solved based on the same technique. A sufficient optimality criteria was given for constrained, concave minimization problems. The precise mathematics of the model transformation from high-level models to Petri nets was defined, and automatic model transformations were carried out to realize these transformations: a transformation from UML models given by the GRM profile to Petri nets and a general algorithm that delivers models to diagnose from UML models. The conformancy of these models to standards was investigated. The probabilistic diagnosis problem in multiprocessor systems was solved using PNS techniques. The efficiency of the method was shown. There were efforts to elaborate a test-based approach of diagnostics, and to parameterize diagnostics models based on dependability experiments. Experiments were carried out to transform IBM Holosofx models to Petri nets

Alpha-MSH induces vasodilatation and exerts cardioprotection via the heme-oxygenase pathway in rat hearts

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Intestinal Microbiota Changes in Mice Lacking Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) - Bifidobacteria Make the Difference

Pituitary adenylate cyclase activating polypetide (PACAP) constitutes a neuropeptide that is widely distributed in the host exerting essential cytoprotective properties, whereas PACAP-/- mice display increased susceptibility to distinct immunopathological conditions. The orchestrated interplay between the gut microbiota and the host is pivotal in immune homeostasis and resistance to disease. Potential pertubations of the intestinal microbiota in PACAP-/- mice, however, have not been addressed so far. For the first time, we performed a comprehensive survey of the intestinal microbiota composition in PACAP-/- and wildtype (WT) mice starting 2 weeks postpartum until 18 months of age applying quantitative culture-independent techniques. Fecal enterobacteria and enterococci were lower in PACAP-/- than WT mice aged 1 month and >/=6 months, respectively. Whereas Mouse Intestinal Bacteroides were slightly higher in PACAP-/- versus WT mice aged 1 and 6 months, this later in life held true for Bacteroides/Prevotella spp. (>/=12 months) and lactobacilli (>15 months of age). Strikingly, health-beneficial bifidobacteria were virtually absent in the intestines of PACAP-/- mice, even when still breastfed. In conclusion, PACAP deficiency is accompanied by distinct changes in fecal microbiota composition with virtually absent bifidobacteria as a major hallmark that might be linked to increased susceptibility to disease

Pituitary adenylate cyclase-activating polypeptide ameliorates vascular dysfunction induced by hyperglycaemia

BACKGROUND: Short-lasting hyperglycaemia occurs frequently in prediabetes and poorly controlled diabetes mellitus leading to vascular damage. Pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to play a protective role in vascular complications of diabetes; moreover, antioxidant effects of PACAP were also described. Therefore, we hypothesized that PACAP exerts protective effects in short-term hyperglycaemia-induced vascular dysfunctions. METHODS: After short-term hyperglycaemia, acetylcholine-induced and sodium nitroprusside-induced vascular relaxation of mouse carotid arteries were tested with a myograph with or without the presence of PACAP or superoxide dismutase. Potential direct antioxidant superoxide-scavenging action of pituitary adenylate cyclase-activating peptide was tested with pyrogallol autoxidation assay; furthermore, the effect of pituitary adenylate cyclase-activating peptide or superoxide dismutase was investigated on hyperglycaemia-associated vascular markers. RESULTS: PACAP administration resulted in reduced endothelial dysfunction after a 1-h hyperglycaemic episode. PACAP was able to restore acetylcholine-induced relaxation of the vessels and improved sodium nitroprusside-induced relaxation. This effect was comparable to the protective effect of superoxide dismutase, but PACAP was unable to directly scavenge superoxide produced by autoxidation of pyrogallol. Endothelial dysfunction was associated with elevated levels of fibroblast growth factor basic, matrix metalloproteinase 9 and nephroblastoma overexpressed gene proteins. Their release was reduced by PACAP administration. CONCLUSION: These results suggest a strong protective role of PACAP in the vascular complications of diabetes

VPAC1 receptors play a dominant role in PACAP-induced vasorelaxation in female mice

PACAP and VIP are closely related neuropeptides with wide distribution and potent effect in the vasculature. We previously reported vasomotor activity in peripheral vasculature of male wild type (WT) and PACAP-deficient (KO) mice. However, female vascular responses are still unexplored. We hypothesized that PACAP-like activity is maintained in female PACAP KO mice and the mechanism through which it is regulated differs from that of male PACAP KO animals.We investigated the vasomotor effects of VIP and PACAP isoforms and their selective blockers in WT and PACAP KO female mice in carotid and femoral arteries. The expression and level of different PACAP receptors in the vessels were measured by RT-PCR and Western blot.In both carotid and femoral arteries of WT mice, PACAP1-38, PACAP1-27 or VIP induced relaxation, without pronounced differences between them. Reduced relaxation was recorded only in the carotid arteries of KO mice as compared to their WT controls. The specific VPAC1R antagonist completely blocked the PACAP/VIP-induced relaxation in both arteries of all mice, while PAC1R antagonist affected relaxation only in their femoral arteries.In female WT mice, VPAC1 receptors appear to play a dominant role in PACAP-induced vasorelaxation both in carotid and in femoral arteries. In the PACAP KO group PAC1R activation exerts vasorelaxation in the femoral arteries but in carotid arteries there is no significant effect of the activation of this receptor. In the background of this regional difference, decreased PAC1R and increased VPAC1R availability in the carotid arteries was found

Continuous-flow synthesis of 3,5-disubstituted pyrazoles via sequential alkyne homocoupling and Cope-type hydroamination

A flow chemistry-based approach is presented for the synthesis of 3,5-disubstituted pyrazoles via sequential copper-mediated alkyne homocoupling and Cope-type hydroamination of the intermediary 1,3-diynes in the presence of hydrazine as nucleophilic reaction partner. The proposed multistep methodology offers an easy and direct access to valuable pyrazoles from cheap and readily available starting materials and without the need for the isolation of any intermediates

Structural and Morphometric Comparison of Lower Incisors in PACAP-Deficient and Wild-Type Mice

Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide with widespread distribution. PACAP plays an important role in the development of the nervous system, it has a trophic and protective effect, and it is also implicated in the regulation of various physiological functions. Teeth are originated from the mesenchyme of the neural crest and the ectoderm of the first branchial arch, suggesting similarities with the development of the nervous system. Earlier PACAP-immunoreactive fibers have been found in the odontoblastic and subodontoblastic layers of the dental pulp. Our previous examinations have shown that PACAP deficiency causes alterations in the morphology and structure of the developing molars of 7-day-old mice. In our present study, morphometric and structural comparison was performed on the incisors of 1-year-old wild-type and PACAP-deficient mice. Hard tissue density measurements and morphometric comparison were carried out on the mandibles and the lower incisors with micro-CT. For structural examination, Raman microscopy was applied on frontal thin sections of the mandible. With micro-CT morphometrical measurements, the size of the incisors and the relative volume of the pulp to dentin were significantly smaller in the PACAP-deficient group compared to the wild-type animals. The density of calcium hydroxyapatite in the dentin was reduced in the PACAP-deficient mice. No structural differences could be observed in the enamel with Raman microscopy. Significant differences were found in the dentin of PACAP-deficient mice with Raman microscopy, where increased carbonate/phosphate ratio indicates higher intracrystalline disordering. The evaluation of amide III bands in the dentin revealed higher structural diversity in wild-type mice. Based upon our present and previous results, it is obvious that PACAP plays an important role in tooth development with the regulation of morphogenesis, dentin, and enamel mineralization. Further studies are required to clarify the molecular background of the effects of PACAP on tooth development

Examination of PACAP-Like Immunoreactivity in Urogenital Tumor Samples.

Numerous studies investigated the localization of pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors in different tumors and described the effects of analogs on tumor growth to show its potential role in oncogenesis. Recently, our research group has found significantly lower levels of PACAP27-like immunorreactivity (LI) and PACAP38-LI in different human samples of primary small cell lung cancer and colon cancer compared to normal healthy tissues. There are only few human studies showing the presence of PACAP and its receptors in urogenital tumors; therefore, the aim of the present study was to compare PACAP-LI in different healthy and pathological human samples from urogenital organs (kidney, urinary bladder, prostate, testis) with radioimmunoassay (RIA) method. Similar to our earlier observations, the PACAP27-LI was significantly lower compared to PACAP38-LI in all samples. We did not find significant alterations in PACAP-LI between healthy and tumoral samples from the urinary bladder and testis. On the other hand, we found significantly lower PACAP38-LI level in kidney tumors compared with healthy tissue samples, and we showed higher PACAP27-LI in prostatic cancer compared to samples from benign prostatic hyperplasia. These data indicate that PACAP levels of different tissue samples are altered under pathological conditions suggesting a potential role of PACAP in the development of different urogenital tumors