Tato diplomová práce se věnuje tématu nehmotného majetku. V první části je porovnávána úprava této problematiky v české legislativě se standardem IAS 38. Text je doplněn řadou příkladů a zobrazení dané problematiky v účetnictví. V závěru teoretické části je zmíněn i daňový pohled a také zobrazení nehmotného majetku podle US GAAP. Praktická část byla vytvořena ve spolupráci se společností ZENTIVA a.s., která poskytla potřebné materiály a zodpověděla dotazy. Na konci diplomové práce je uveden seznam pojmů, které se často vyskytují v této problematice.

La parodontite est une maladie inflammatoire chronique prévalente due à la réponse de l'hôte aux bactéries orales telles que Porphyromonas Gingivalis, un pathogène clé. L’IL-1β, l’IL-6, le TNF-α, l’IL-17A sont des cytokines pro-inflammatoires cruciales dans la pathogenèse de la parodontite qui perpétuent l'inflammation gingivale et la résorption osseuse alvéolaire. Dans les parodontites réfractaires aux traitements conventionnels qui visent à réduire le stimulus bactérien, les thérapies qui modulent la réponse de l’hôte en bloquant ces cytokines proinflammatoires ne sont que partiellement efficaces. Il a été suggéré que d'autres membres de la famille de l’IL- 1, les cytokines IL-33, et IL-36 (IL-36α/IL-36γβ/IL- 36βγ/IL-36Ra/IL-38) pourraient présenter un intérêt dans la parodontite. Ce travail vise à analyser l'expression et le rôle de ces interleukines dans la destruction osseuse alvéolaire associée à la parodontite pour déterminer si elles pourraient constituer des cibles thérapeutiques intéressantes. Nos données suggèrent l’implication de l’IL-33 et des IL-36 (l’IL-36γ en particulier) dans la pathogenèse de la parodontite. L’IL-33 et IL-36γ semblent jouer un rôle central dans la réponse immunitaire innée au stimulus bactérien et pourrait favoriser l'ostéoclastogenèse en augmentant l’expression du RANKL dans les cellules gingivales. Les nouveaux traitements bloquant ces cytokines pourraient être prometteurs dans la parodontite. / Periodontitis is a prevalent chronic inflammatory disease due to the host response to oral bacteria such as Porphyromonas Gingivalis, a keystone pathogen. IL-1β, IL-6, TNF-α, IL-17A are crucial proinflammatory cytokines in the pathogenesis of periodontitis perpetuating the gingival inflammation and the alveolar bone resorption. In periodontitis refractory to conventional treatment which aims at reducing the bacterial challenge, host modulation therapies blocking these pro-inflammatory cytokines are only partially effective. It has been suggested that other members of the IL-1 family, IL-36 cytokines (IL-36α/IL-36β/IL-36γ/IL- 36Ra/IL-38) could be of interest in periodontitis. The objective of this thesis is to investigate the expression and role of these interleukin in the alveolar bone resorption associated to periodontitis to determine if they could represent interesting therapeutic targets. Our findings suggest an involvement of IL-33 and IL-36 (IL-36γ in particular) in the pathogenesis of periodontitis. IL-33 and IL-36γ seems to play a pivotal role in the innate immune response to bacterial challenge and could support osteoclastogenesis by enhancing RANKL expression in gingival cells. Novel therapeutics blocking these cytokines could be promising in periodontitis.

<p><strong>Sammanfattning</strong></p><p>Olikheter mellan länder och deras olika regelverk gör att företags års-redovisningar skiljer sig åt och intressenter får svårare att jämföra finansiell information. Olikheter leder i sin tur till olika definitioner och värderingar vid redovisning, vilket gör att jämförbarheten mellan företag och värdering av ett företag blir en komplicerad verksamhet. (FAR 2009). Immateriella tillgångar har diskuterats flitigt och det finns argument både för och emot huruvida företagen skall aktivera tillgången i balansräkningen (Sundgren et al 2007). FoU inom läkemedelsindustrin är en mycket riskfylld verksamhet, eftersom endast cirka 10 procent av alla läkemedel som når utvecklingsfasen kommer ut på marknaden till slut och cirka 20 procent av de marknadsförda läkemedlen är tillräckligt lönsamma för att så småningom täcka den investering som gjorts i dem (Nickisch et al 2009).</p><p>Undersökningen ämnar svara på följande frågor: Vilka är de huvudsakliga faktorer som begränsar företagen vid redovisning av FoU? I vilken omfattning aktiverar företagen kostnader för FoU som en tillgång? Vilka är de tänkbara orsakerna bakom läkemedelsföretagens val av redovisning av FoU?</p><p>Syftet med uppsatsen är att identifiera hur företagen inom läkemedels-branschen redovisar FoU i sina räkenskaper, samt vilka begränsningar och möjligheter företagen står inför i och med gällande regelverk och rekommen-dationer.</p><p>Uppsatsen är i huvudsak konstruerad med hjälp av den kvalitativa metoden. Dock har den även inslag av kvantitativ metod, eftersom omfattningen av tillgångsföring av kostnader för FoU inom läkemedelsindustrin är av stort intresserade.</p><p>För att skapa en rimlig omfattning på problemet och på undersökningen i övrigt avgränsades undersökningen till att omfatta svenska börsnoterade företag inom läkemedelsbranschen forskning, utveckling och tillverkning i egen regi.</p><p>I teorikapitlet beskrivs historik avseende det regelverk som läkemedelsföretagen redovisar enligt. Dessutom presenteras aktuella regelverk i dagsläget, teorier kring risk, värdering och läkemedelsindustrin samt de faser som framställningen av ett läkemedel kan befinna sig i.</p><p>Företagen tenderar till att vara mycket försiktiga vid redovisning av FoU. Kostnader för FoU aktiveras i så gott som obefintlig omfattning.</p>

Sammanfattning Olikheter mellan länder och deras olika regelverk gör att företags års-redovisningar skiljer sig åt och intressenter får svårare att jämföra finansiell information. Olikheter leder i sin tur till olika definitioner och värderingar vid redovisning, vilket gör att jämförbarheten mellan företag och värdering av ett företag blir en komplicerad verksamhet. (FAR 2009). Immateriella tillgångar har diskuterats flitigt och det finns argument både för och emot huruvida företagen skall aktivera tillgången i balansräkningen (Sundgren et al 2007). FoU inom läkemedelsindustrin är en mycket riskfylld verksamhet, eftersom endast cirka 10 procent av alla läkemedel som når utvecklingsfasen kommer ut på marknaden till slut och cirka 20 procent av de marknadsförda läkemedlen är tillräckligt lönsamma för att så småningom täcka den investering som gjorts i dem (Nickisch et al 2009). Undersökningen ämnar svara på följande frågor: Vilka är de huvudsakliga faktorer som begränsar företagen vid redovisning av FoU? I vilken omfattning aktiverar företagen kostnader för FoU som en tillgång? Vilka är de tänkbara orsakerna bakom läkemedelsföretagens val av redovisning av FoU? Syftet med uppsatsen är att identifiera hur företagen inom läkemedels-branschen redovisar FoU i sina räkenskaper, samt vilka begränsningar och möjligheter företagen står inför i och med gällande regelverk och rekommen-dationer. Uppsatsen är i huvudsak konstruerad med hjälp av den kvalitativa metoden. Dock har den även inslag av kvantitativ metod, eftersom omfattningen av tillgångsföring av kostnader för FoU inom läkemedelsindustrin är av stort intresserade. För att skapa en rimlig omfattning på problemet och på undersökningen i övrigt avgränsades undersökningen till att omfatta svenska börsnoterade företag inom läkemedelsbranschen forskning, utveckling och tillverkning i egen regi. I teorikapitlet beskrivs historik avseende det regelverk som läkemedelsföretagen redovisar enligt. Dessutom presenteras aktuella regelverk i dagsläget, teorier kring risk, värdering och läkemedelsindustrin samt de faser som framställningen av ett läkemedel kan befinna sig i. Företagen tenderar till att vara mycket försiktiga vid redovisning av FoU. Kostnader för FoU aktiveras i så gott som obefintlig omfattning.

Zugl.: Bochum, Universiẗat, Diss., 2004.

Les IL-36α, β et γ sont des cytokines appartenant à la famille de l’IL-1. Elles sont exprimées notamment dans la peau et sont impliquées dans la pathogenèse du psoriasis. Leurs inhibiteurs connus ou hypothétiques l’IL-36Ra et l’IL-38 agissent en limitant l’inflammation. Dans la polyarthrite rhumatoïde (PR) et la maladie de Crohn, l’expression et le rôle de ces cytokines sont encore débattues. Les travaux de cette thèse se consacrent à l’étude du profil d’expression des IL-36 et de leurs inhibiteurs ainsi qu’à déterminer l’impact d’une surexpression de l’IL-38 in vivo et in vitro. La première étude a été réalisée chez les patients atteints de PR en comparaison avec les patients psoriasiques et ceux atteints de maladie de Crohn ainsi que dans les modèles murins correspondants. Cette étude a montré que ces cytokines étaient exprimées majoritairement par les kératinocytes et les macrophages mais possédaient cependant des profils d’expression distincts. Elle a également permis d’identifier un sous-groupe de patients pour lesquelles les IL-36 pourraient avoir un rôle important et pourraient représenter une cible en clinique. La seconde partie de la thèse a permis de montrer un effet anti-inflammatoire d’une surexpression de l’inhibiteur IL-38 dans différents modèles d’arthrite in vivo et dans des macrophages in vitro. Les thérapies basées sur l’inhibition de cytokines ont déjà prouvé leur efficacité en clinique et sont une cible thérapeutique très prometteuse. Cette thèse a pour objectifs d’une part de mieux comprendre la biologie des nouvelles cytokines de la famille de l’IL-1 et d’autre part de participer à la découverte de nouvelles cibles thérapeutiques dans les pathologies inflammatoires chroniques telles que la PR. / IL-36α, β and γ are cytokines belonging to the IL-1 family. IL- 36 are expressed especially in the skin and are involved in the pathogenesis of psoriasis. Their inhibitors (well-known or hypothetical) IL-36Ra and IL-38 act in reducing inflammation. In rheumatoid arthritis (RA) and Crohn's disease, the expression and role of these cytokines need to be elucidated. This thesis is dedicated to the study of IL-36 agonists and antagonists expression profile and the better understanding of in vivo and in vitro impact of IL-38 overexpression. The firts study was conducted in patients with RA in comparison with psoriasis and Crohn's disease patients as well as in the corresponding relevant murine models. This study showed that these cytokines were expressed predominantly by keratinocytes and macrophages but had distinct expression profiles. A subgroup of patients for which the IL-36 could have an important role and may represent a clinical target was also identified. In the second part of the thesis, we demonstrated an anti-inflammatory effect following overexpression of the inhibitor IL-38 in several in vivo arthritis models and in macrophages in vitro. Cytokine neutralization based therapies have already proven effective in the clinic and are still a promising therapeutic target. This thesis aims firstly to better understand the biology of new IL-1 family cytokines and also to participate in the discovery of new therapeutic targets in chronic inflammatory diseases such as RA.

Pediatric acute myeloid leukemia (AML) is a molecularly heterogeneous disease that arises from genetic alterations in pathways that regulate self-renewal and myeloid differentiation. While the majority of patients carry recurrent chromosomal translocations, almost 20% of childhood AML do not show any recognizable cytogenetic alteration and are defined as cytogenetically normal (CN)-AML. CN-AML patients have always showed a great variability in response to therapy and overall outcome, underlining the presence of unknown genetic changes, not detectable by conventional analyses, but relevant for pathogenesis, and outcome of AML. The development of novel genome-wide techniques such as next-generation sequencing, have tremendously improved our ability to interrogate the cancer genome. Based on this background, the aim of this research study was to investigate the mutational landscape of pediatric CN-AML patients negative for all the currently known somatic mutations reported in AML through whole-transcriptome sequencing (RNA-seq). RNA-seq performed on diagnostic leukemic blasts from 19 pediatric CN-AML cases revealed a considerable incidence of cryptic chromosomal rearrangements, with the identification of 21 putative fusion genes. Several of the fusion genes that were identified in this study are recurrent and might have a prognostic and/or therapeutic relevance. A paradigm of that is the CBFA2T3-GLIS2 fusion, which has been demonstrated to be a common alteration in pediatric CN-AML, predicting poor outcome. Important findings have been also obtained in the identification of novel therapeutic targets. On one side, the identification of NUP98-JARID1A fusion suggests the use of disulfiram; on the other, here we describe alteration-activating tyrosine kinases, providing functional data supporting the use of tyrosine kinase inhibitors to specifically inhibit leukemia cells. This study provides new insights in the knowledge of genetic alterations underlying pediatric AML, defines novel prognostic markers and putative therapeutic targets, and prospectively ensures a correct risk stratification and risk-adapted therapy also for the “all-neg” AML subgroup.

L’impatto del microbiota intestinale(MI) sulla mortalità correlata al trapianto allogenico di cellule staminali emopoietiche(TCSE) è stato recentemente dimostrato. Questa osservazione corrobora l’idea di un ruolo significativo del MI nella ricostruzione immunologica successiva al TCSE e nella genesi della Graft-versus-Host-Disease acuta(GvHD). Abbiamo pertanto condotto il primo studio longitudinale sul ruolo del MI nella genesi di GvHD in pazienti pediatrici sottoposti a TCSE. Sono stati arruolati 10pazienti, di cui 5 con GvHD. Per ogni paziente sono stati raccolti campioni fecali seriati ogni 10-15 giorni fino a 100 giorni dopo il TCSE. Il profilo filogenetico del MI è stato caratterizzato mediante pyrosequencing 454 della regione ipervariabile V4 della subunità 16S dell’rRNA. Il profilo funzionale è stato valutato mediante l’analisi degli acidi-grassi-a-corta-catena utilizzando la gas cromatografia-spettroscopia di massa. Dopo il TCSE è stata osservata una profonda distruzione strutturale e funzionale del normale assetto mutualistico dell’ecosistema intestinale. La traiettoria di ricostruzione del MI dopo il TCSE è risultata essere significativamente differente tra i pazienti con e senza GvHD. In particolare, nei pazienti senza GvHD è stata evidenziata prima del TCSE una precisa signature del MI, caratterizzata da un’elevata concentrazione di Bacteroidetes e Parabacteoidetes(p<0.05, Fig. 1). Parallelamente nei pazienti senza GVHD è stato osservato un aumento significativo degli acidi-grassi-a-corta-catena e di propionato in particolare(p<0.05). Questa caratteristica signature si è proiettata dopo il TCSE, persistendo alla distruzione dell’ecosistema intestinale e dimostrando l’elevata adattabilità di questi germi. I nostri dati indicano che le dinamiche dell’ecosistema microbico intestinale possono essere un fattore in grado di influenzare l’insorgenza di GvHD. In particolare, la presenza di un profilo mutualistico pre-TCSE del MI, caratterizzato dalla presenza di germi produttori di acidi-grassi-a-corta-catena con riconosciute proprietà immunomodulatorie, sembra mitigare il rischio di sviluppare GVHD. Questi risultati aprono quindi nuove prospettive sulla possibilità di manipolare il MI pre-TCSE per modulare la ricostruzione del sistema immunitario. / The impact of the gut microbiota(MI) on allogeneic hematopoietic stem cell transplantation(HSCT) has been recently demonstrated. This observation supported the idea of a significant role of MI in the immunological reconstruction after HSCT and in the genesis of acute Graft-versus-Host-Disease(GvHD). We therefore conducted the first longitudinal study on the role of MI in the onset of GvHD in pediatric patients undergoing HSCT. 10 patients were enrolled, including 5 with GvHD. For each patient were collected seriated fecal samples every 10-15 days up to 100 days after HSCT. The phylogenetic profile of MI was characterized by pyrosequencing 454 of the hypervariable V4 region of the 16S rRNA subunit. The functional profile was evaluated by analysis of fatty acid-fat-to-short-chain using the gas chromatography-mass spectroscopy. After HSCT a profound structural and functional destruction of the normal position mutualistic gut ecosystem was observed. The reconstruction of the trajectory of the MI after HSCT was significantly different between patients with and without GVHD. In particular, in patients without GVHD, a precise of the MI signature before the HSCT was highlighted, characterized by a high concentration of Bacteroidetes and Parabacteoidetes(p <0.05, Fig. 1). In parallel in patients without GVHD it was observed a significant increase of the fatty-acid-to-short-chain and in particular propionate(p <0.05). This characteristic signature was projected after HSCT, persisting to the intestinal ecosystem destruction and demonstrating the high adaptability of these germs. Our data indicate that the intestinal microbial ecosystem dynamics can be a factor influencing the onset of GvHD. In particular, the presence of a mutualistic profile pre-HSCT of the MI, characterized by the presence of germs producers of acid-fat-to-short-chain recognized with immunomodulatory, seems to mitigate the risk of developing GVHD. These results therefore open new perspectives on the possibility of manipulating the MI pre-HSCT to modulate the reconstruction of the immune system.

Background. L’ asfissia perinatale determina spesso un quadro di encefalopatia associata a disfunzione cardiaca. L’ipotermia rappresenta l’unica terapia efficace nel ridurre il danno cerebrale. Non è noto se possa agire anche sul danno miocardico. Obiettivi. Definire il danno miocardico conseguente ad asfissia perinatale mediante metodiche TDI associate a tecniche ecocardiografiche tradizionali e valutarne l’ outcome a breve e lungo termine. Materiali e Metodi. Sono stati arruolati 15 neonati con encefalopatia ipossico-ischemica: 7 di grado severo-moderato raffreddati per 72 h e 8 di grado lieve non raffreddati. Entrambi i gruppi hanno eseguito dosaggio ematico di CPK, Troponina T e Pro-BPN entro le 6 h di vita, a 12, 24 h di vita e in 7° giornata; a tutti sono stati eseguiti ECG ed ecocardiografia tradizionale e TDI entro 6 h di vita, a 36 h, a 7 giorni ed a 12 mesi; nel gruppo 1 anche a 4 giorni e a 1, 6 e 18 mesi di vita. Risultati. Le concentrazioni sieriche dei markers studiati nelle diverse tempistiche sono risultate più elevate nei pazienti del gruppo 1 rispetto al gruppo 2. In maniera concorde tutti gli indici di funzionalità cardiaca (output ventricolari, TAPSE, velocità sistolica e diastoliche al TDI) sono risultati significativamente inferiori nel gruppo1 rispetto al gruppo2 nelle prime ore dopo l’ insulto ischemico per entrambi i ventricoli, per poi riallinearsi successivamente. In particolare per quanto riguarda la metodica TDI i rapporti E/Em ed Em/Am per la valvola mitrale sono risultati significativamente inferiori anche a 7 giorni nel gruppo 1. Conclusioni. Le misure eseguite con metodica TDI presentano maggiore sensibilità nel riconoscere alterazioni della funzionalità miocardica rispetto alle metodiche tradizionali; tali valori infatti si normalizzano più tardivamente. Riteniamo pertanto opportuno valutare la funzionalità miocardica almeno per alcuni giorni dopo il riscaldamento nei bambini affetti da encefalopatia moderato-severa sottoposti ad ipotermia. / Background. Perinatal asphyxia is commonly associated with hypoxic ischaemic encephalopathy and cardiovascular dysfunction. Therapeutic hypothermia has become standard treatment for moderate and severe neonatal hypoxic–ischemic encephalopathy to reduce cerebral morbidity and mortality. The effect on the heart is incompletely explored. Aim. To assess myocardial performance of infants with perinatal asphyxia using traditional and DTI echocardiographycs technique; evaluation of early and long-term myocardial outcome. Study design. Fifteen infants with hypoxic ischaemic encephalopathy were enrolled in the study: seven infants with moderate-severe hypoxic ischaemic encephalopathy cooled for 72 hours (group1); eight normothermic infants with mild hypoxic ischaemic encephalopathy (group2).
Biomarkers serum concentrations (CPK,TroponinT, PRO-BNP) were measured at 6, 12 , 24 h of life and on the seven day of life. ECG and echocardiographies (traditional and TDI mesaurements) were done in the first 6 h of life, at 36h, on day 7, at 12 month. In the cooled infants were done also on day 4, at 1, 6 and 18 month. Results. All sieric biomarkers were always higher in the group1 than group2. All functional myocardial indices (ventricular output, TAPSE, systolic velocity (Sm), early diastolic velocity (Em), late diastolic velocity (Am)) were lower for both ventricles in the first hours after hypoxic insult in the group1. Moreover we have also found that mitral E/Em ratio and Em/Am ratio were lower in the group 1 on day 7. Conclusion. The DTI technique appears more sensitive than traditional echocardiography to the subtle changes in cardiac performance that occur after perinatal asphyxia. In particular these value were lower for more time that conventional parameters. Moreover, in cooled infants with moderate-severe hypoxic ischaemic encephalopathy, it may be important examinated myocardial function at least for a few days after rewarming.

Le “RASopatie” includono un gruppo di patologie congenito-malformative a trasmissione autosomica-dominante causate da mutazioni eterozigoti germinali in geni che codificano per proteine del pathway RAS-MAPKinasi. Sono caratterizzate da dismorfismi faciali, iposomia, cardiopatia congenita, anomalie ectodermiche e scheletriche, coinvolgimento cognitivo e suscettibilità tumorale. La bassa statura è uno dei principali elementi distintivi sul quale si può agire sotto il profilo terapeutico, mediante somministrazione di Ormone della Crescita biosintetico (GH). In questo studio è stato analizzato l’andamento accrescitivo in un ampio gruppo di 88 pazienti affetti da Rasopatia con diagnosi molecolare confermata. Sono stati valutati: distribuzione per genotipo, prevalenza delle caratteristiche fenotipiche e correlazione genotipo-fenotipo sull’intero gruppo di studio ed in particolare andamento accrescitivo spontaneo, proporzioni corporee, sviluppo puberale, e dati di statura finale (FH) in 33 soggetti, di cui 16 trattati con GH per deficit secretivo (GHD). Valutati inoltre efficacia e sicurezza della terapia con GH, definito il rischio oncologico e quantificato il coinvolgimento psico-cognitivo. 33 pazienti hanno mostrato GHD e sono stati trattati con GH per 6,8 ± 4,8 anni. Prima dell’ inizio della terapia, la velocità di crescita era - 2.6 ± 1.3 SDS e i livelli basali di IGF1 pari a - 2.0 ± 1.1 SDS. La terapia con GH a lungo termine, iniziata precocemente durante l’infanzia, ha determinato un guadagno staturale positivo rispetto ai pazienti non trattati (+1,3 SDS), normalizzando la FH per gli standards di condizione ma non per la popolazione generale e il Target staturale parentale. Il timing di sviluppo puberale ha influenzato negativamente lo scatto di crescita puberale. 1/88 soggetti, non sottoposta a terapia con GH, ha sviluppato neoplasia. Sono emersi ritardo mentale (41.1%), deficit visuo-spaziali (41.8%), ADHD (38.8%), anomalie del SNC (24.4%) e anomalie del tracciato elettroencefalografico (6.7%). / RASopathies are developmental disorders caused by heterozygous germline mutations in genes encoding proteins in the RAS-MAPK signaling pathway. These conditions share facial dysmorphism, failure to thrive, congenital heart disease, ectodermal, and skeletal anomalies, variable cognitive involvement, and susceptibility to certain malignancies as major characteristics. This study analized clinical features, growth trend and body proportions in 88 patients affected by RASopathies with molecularly confirmed diagnosis, and FH reached in 33, including 16 treated with GH therapy for proven GH deficiency. Clinically, 69 patients (78.4%) had a diagnosis of Noonan syndrome (NS), seven of NS/Loose anagen hair (8.0%), six of CFC syndrome (6.8%), and two of Costello syndrome, Noonan syndrome with multiple lentigines (NSML) and Legius syndrome. Among them, 52 (59.1%) had PTPN11 mutations, while the other genotypes were less than 10%. Most patients were born at term, after a physiological pregnancy and presented regular neonatal adaptation. 75.9% patients had cardiac anomalies and 20.2% needed surgery for cardiac involvement. Thirty-three patients showed GH deficiency after pharmacological tests, and were GH-treated for an average period of 6.8±4.8 years. Before starting therapy, HV was -2.6±1.3 SDS, and mean basal IGF1 levels were -2.0±1.1 SDS. Long-term GH therapy, starting early during childhood, resulted in a positive height response compared with untreated patients (1.3 SDS in terms of height-gain), normalizing FH for Ranke standards but not for general population and Target Height. The delayed pubertal development and the inadequate pubertal catch-up growth could explain the impaired FH. Our patients on GH-therapy benefitted from the pharmacological treatment if started in pre-puberty and given for a long time. Probably, the prepubertal start of GH-treatment could compensate the lack of a pubertal growth. 1/88 patient had cancer, but she didn’t received GH. The patients presented cognitive impairment (41.1%), ADHD (38.8%), SNC anomalies (24.4%) and EEG alterations (6.7%).