Combining SOA and BPM Technologies for Cross-System Process Automation

This paper summarizes the results of an industry case study that introduced a cross-system business process automation solution based on a combination of SOA and BPM standard technologies (i.e., BPMN, BPEL, WSDL). Besides discussing major weaknesses of the existing, custom-built, solution and comparing them against experiences with the developed prototype, the paper presents a course of action for transforming the current solution into the proposed solution. This includes a general approach, consisting of four distinct steps, as well as specific action items that are to be performed for every step. The discussion also covers language and tool support and challenges arising from the transformation

Bringing Model Checking Closer To Practical Software Engineering

Bal, H.E. [Promotor]Templon, J.A. [Copromotor]Willemse, T.A.C. [Copromotor

Telomere length in cardiovascular disease and type 2 diabetes

Telomere attrition during mitosis is thought to be a mechanism for cell senescence, which is induced when the mean length reduces below a critical value. Oxidative DNA damage has been suggested to cause greater telomere loss per cell division. Thus telomere length indicates the cells' replicative capacity and "biological age". Premature tissue ageing and senescence are major features of cardiovascular disease (CVD) and type 2 diabetes (T2D), and as such telomere length might play an important role in their pathogenesis. The aim of this thesis was to explore the association of telomere length with CVD and type 2 diabetes (T2D) as well as the CVD/T2D risk factors determining telomere length. The present study showed that telomere length is shorter in CVD and T2D patients compared to healthy subjects. As to what determines telomere length, I found that at least in the case of CVD, it can in part be partly attributed to inherited short telomeres, expressing the family history of the disease. At the same time, in diabetes patients I have observed that high oxidative stress, probably induced by the disease risk factors in the patients, is associated with greater telomere shortening. The later was consistent with the association found between variation in genes regulating reactive oxygen species levels, and shorter telomeres. I further examined the possible determinants of telomere shortening with in vitro experiments on ageing fibroblasts. In this case, the effect of glucose-induced oxidative stress and/or pro-inflammatory conditioning on telomere shortening was tested. The results showed that pro-inflammatory conditioning, partly by inducing an increased cell turnover, aggravates the shortening of telomeres in long-term culture. Telomere length may prove to be very useful in the management and possibly the prediction of CVD and diabetes, representing the contribution to their pathology of age, oxidative stress and chronic inflammation

Cytosolic nucleic acids as new markers of senescence and hyperglycemia-induced inflammation

openAging is characterized by a gradual functional decline resulting from a complex interaction between genetic, epigenetic and stochastic factors and the rate of aging is recognized as the most important risk factor for the development of the most common age-related diseases (ARDs). Research on aging process is currently focused on understanding the molecular mechanisms underlying age-related features accompanying on one hand, the onset of ARDs and on the other hand, the chances to reach a successful aging. A chronic, systemic, low-grade, age-related pro-inflammatory status, called “Inflammaging” is correlated with ARDs development. The two main culprits of inflammaging are the repeated stimulation of immune system over time and the increased burden of senescent cells. Senescent cells are able to modify the microenvironment acquiring a senescence-associated secretory phenotype (SASP). In patients affected by ADRs, the rate of aging process in increased, as well as the burden of senescent cells with SASP. In this scenario, a better understanding of the molecular mechanisms that promote cellular senescence is of basic and clinical relevance. A number of mechanisms promoting senescence have been well characterized. Increasing evidence suggest that nuclear DNA fragility and a sub-functional DNA damage response (DDR) is associated with an increase release of nucleic acids in cytoplasm, including RNA:DNA hybrids. However, only few studies have tempted to assess the pro- or anti-inflammatory effects of this misplaced nucleic acid pool in different cellular models. In this framework, we have analysed the cytoplasmic pool of misplaced nucleic acids in a model of human endothelial cells (HUVECs), in two prototypical stress conditions related to inflammaging, such as “replicative cellular senescence” and “hyperglycemic condition”. A significant increased amount of misplaced nucleic acids was observed in the cytoplasm of senescent cells compared to the younger ones, and in young cells cultured for 1-week in hyperglycemic condition compared to young cells cultured in normoglycemic medium. The cytosolic pool of misplaced nucleic acids is composed of dsDNA, including micronuclei, bubbles and telomere sequences, dsRNA and hybrids. A reduced expression of RNaseH2, the enzyme involved in the degradation of RNA moiety of RNA:DNA hybrids was observed in senescent cells compared to the younger ones and in young cells treated with hyperglycemic medium. The amounts of mis incorporated ribonucleotides, biomarkers od genome instability, were inversely related to RNaseH2 expression level, suggesting that both “senescence” and “hyperglycemic condition” are associated with increased genomic instability. Since the presence of nucleic acids in cytoplasm can activate a number of cytosolic receptors inducing the antiviral response, characterized by increased release of type 1 Interferon (IFN-1), we analysed cGAS/STING/IRF3 axis and IFN-1 expression in “senescence” and “hyperglycemic condition”. Surprisingly, a strong reduction of cGAS expression was observed in senescent cells in normo and hyperglycemic conditions, in association with the absence of IFN-1 modulation, whereas the expression of proinflammatory cytokines, such as IL-1beta, IL-6 and IL-8 was significantly increased. Overall, our results suggest an imbalance between antiviral and proinflammatory response in senescent cells and in young cells under hyperglycemic conditions. Increasing evidence suggest that senescent cells have an enhanced susceptibility to viral infections. We hypothesized that such imbalance between antiviral and proinflammatory responses could be, almost in part, a direct consequence of their ability “to tolerate” a high cytosolic nucleic acids load. Overall, our results pave the way to explain why old subjects, especially the elderly patients affected by diabetes, are more susceptible to adverse outcomes of infectious diseases.L'invecchiamento è caratterizzato da un graduale declino funzionale derivante da una complessa interazione tra fattori genetici e epigenetici insieme ad un'aumentata velocità di invecchiamento, riconosciuta come il fattore di rischio più importante per lo sviluppo delle malattie associate all'età più comuni (ARD). La ricerca sul processo di invecchiamento è attualmente focalizzata sulla comprensione dei meccanismi molecolari alla base delle caratteristiche legate all'età che accompagnano, da un lato, l'insorgenza di ARD e dall'altro, le possibilità di raggiungere un invecchiamento di successo. Uno stato pro-infiammatorio cronico, sistemico, di basso grado e legato all'età, chiamato "Inflammaging" è correlato allo sviluppo degli ARD. I due principali responsabili dell'inflammaging sono la stimolazione ripetuta del sistema immunitario nel tempo e l'aumento del numero di cellule senescenti. Le cellule senescenti sono in grado di modificare il microambiente acquisendo un fenotipo secretorio associato alla senescenza (SASP). Nei pazienti affetti da ADR, la velocità del processo d’invecchiamento è aumentato, così come il numero delle cellule senescenti con SASP. In questo scenario, una migliore comprensione dei meccanismi molecolari che promuovono la senescenza cellulare è di rilevanza di base e clinica. Prove crescenti suggeriscono che la fragilità del DNA nucleare e una risposta sub-funzionale al danno al DNA (DDR) si associano a un aumento del rilascio di acidi nucleici nel citoplasma, inclusi gli ibridi RNA:DNA. In questa cornice, abbiamo analizzato il pool di acidi nucleici rilasciati nel citoplasma in un modello di cellule endoteliali umane (HUVEC), in due condizioni di stress legate all'inflammaging, come la "senescenza replicativa cellulare " e la "condizione iperglicemica". È stata osservata una quantità significativa di acidi nucleici nel citoplasma delle cellule senescenti rispetto a quelle più giovani e nelle cellule giovani coltivate per 1 settimana in condizioni iperglicemiche rispetto alle giovani cellule coltivate in mezzo normoglicemico. Il pool citosolico di acidi nucleici è composto da dsDNA, tra cui micronuclei, bubbles e sequenze telomeriche, dsRNA e ibridi. Una ridotta espressione di RNasi H2, l'enzima coinvolto nella degradazione della componente a RNA degli ibridi RNA:DNA è stato osservato nelle cellule senescenti rispetto a quelle più giovani e nelle cellule giovani trattate con mezzo iperglicemico. Le quantità di ribonucleotidi mal incorporati, biomarcatori dell’instabilità genomica, sono inversamente correlate al livello di espressione di RNasi H2, suggerendo che sia la "senescenza" che la "condizione iperglicemica" sono associate a una maggiore instabilità genomica. Poiché la presenza di acidi nucleici nel citoplasma può attivare un certo numero di recettori citosolici che inducono la risposta antivirale, caratterizzata da un maggiore rilascio di interferone di tipo 1 (IFN-1), abbiamo analizzato l'asse cGAS/STING/IRF3 e l'espressione IFN-1 in "senescenza" e "condizione iperglicemica". Sorprendentemente, una forte riduzione dell'espressione di cGAS è stata osservata nelle cellule senescenti in condizioni normo e iperglicemiche, in associazione con l'assenza di modulazione di IFN-1, mentre l'espressione di citochine pro-infiammatorie, come IL-1 beta, IL-6 e IL-8 è significativamente aumentata. Nel complesso, i nostri risultati suggeriscono uno squilibrio tra la risposta antivirale e pro-infiammatoria nelle cellule senescenti e nelle cellule giovani in condizioni iperglicemiche. Crescenti evidenze suggeriscono che le cellule senescenti hanno una maggiore suscettibilità alle infezioni virali. Abbiamo ipotizzato che tale squilibrio tra risposte antivirali e pro-infiammatorie potrebbe essere, almeno in parte, una conseguenza diretta della loro capacità di "tollerare" un elevato carico di acidi nucleici citosolici.SALUTE DELL'UOMOopenRamini, Debora