Non-classical ProIL-1beta activation during mammary gland infection is pathogen-dependent but caspase-1 independent

Infection of the mammary gland with live bacteria elicits a pathogen-specific host inflammatory response. To study these host-pathogen interactions wild type mice, NF-kappaB reporter mice as well as caspase-1 and IL-1beta knockout mice were intramammarily challenged with Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The murine mastitis model allowed to compare the kinetics of the induced cytokine protein profiles and their underlying pathways. In vivo and ex vivo imaging showed that E. coli rapidly induced NF-kappaB inflammatory signaling concomitant with high mammary levels of TNF-alpha, IL-1 alpha and MCP-1 as determined by multiplex analysis. In contrast, an equal number of S. aureus bacteria induced a low NF-kappaB activity concomitant with high mammary levels of the classical IL-1beta fragment. These quantitative and qualitative differences in local inflammatory mediators resulted in an earlier neutrophil influx and in a more extensive alveolar damage post-infection with E. coli compared to S. aureus. Western blot analysis revealed that the inactive proIL-1beta precursor was processed into pathogen-specific IL-1beta fragmentation patterns as confirmed with IL-1beta knockout animals. Additionally, caspase-1 knockout animals allowed to investigate whether IL-1beta maturation depended on the conventional inflammasome pathway. The lack of caspase-1 did not prevent extensive proIL-1beta fragmentation by either of S. aureus or E. coli. These non-classical IL-1beta patterns were likely caused by different proteases and suggest a sentinel function of IL-1beta during mammary gland infection. Thus, a key signaling nodule can be defined in the differential host innate immune defense upon E. coli versus S. aureus mammary gland infection, which is independent of caspase-1

Inherited human IRAK-1 deficiency selectively impairs TLR signaling in fibroblasts

Most members of the Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) families transduce signals via a canonical pathway involving the MyD88 adapter and the interleukin-1 receptor-associated kinase (IRAK) complex. This complex contains four molecules, including at least two (IRAK-1 and IRAK-4) active kinases. In mice and humans, deficiencies of IRAK-4 or MyD88 abolish most TLR (except for TLR3 and some TLR4) and IL-1R signaling in both leukocytes and fibroblasts. TLR and IL-1R responses are weak but not abolished in mice lacking IRAK-1, whereas the role of IRAK-1 in humans remains unclear. We describe here a boy with X-linked MECP2 deficiency-related syndrome due to a large de novo Xq28 chromosomal deletion encompassing both MECP2 and IRAK1 Like many boys with MECP2 null mutations, this child died very early, at the age of 7 mo. Unlike most IRAK-4- or MyD88-deficient patients, he did not suffer from invasive bacterial diseases during his short life. The IRAK-1 protein was completely absent from the patient's fibroblasts, which responded very poorly to all TLR2/6 (PAM2CSK4, LTA, FSL-1), TLR1/2 (PAM3CSK4), and TLR4 (LPS, MPLA) agonists tested but had almost unimpaired responses to IL-1β. By contrast, the patient's peripheral blood mononuclear cells responded normally to all TLR1/2, TLR2/6, TLR4, TLR7, and TLR8 (R848) agonists tested, and to IL-1β. The death of this child precluded long-term evaluations of the clinical consequences of inherited IRAK-1 deficiency. However, these findings suggest that human IRAK-1 is essential downstream from TLRs but not IL-1Rs in fibroblasts, whereas it plays a redundant role downstream from both TLRs and IL-1Rs in leukocytes

EU:n tietosuoja-asetus 2016/679 (GDPR) ohjelmistoyrityksessä

Insinöörityössä tavoitteena oli tutustua EU:n tietosuoja-asetukseen 2016/679 sekä selvittää, millaisia vaatimuksia se aiheuttaa työn tilanneessa ohjelmistoyrityksessä ja suunnitella ja toteuttaa osa muutostöistä. Työn tavoitteet suunniteltiin yhdessä tilaajayrityksen kanssa. Työn aikana selvitettiin tietosuoja-asetuksen sisältöä ja sitä, miltä osin se kohdistuu yritykseen ja kuinka asetuksen ehdot käytännössä toteutettaisiin. Selvityksen aikana pyrittiin muodostamaan selkeä kuva siitä, missä asetuksen määrittämissä rooleissa yritys toimii ja mitkä niistä muodostuvat vastuut ovat. Pyrittiin myös muodostamaan käsitys yrityksen asiakkaiden tulevista tarpeista, joiden perusteella yrityksen ohjelmistotuote voitaisiin valmistella vastaamaan asetuksen vaatimuksia. Selvityksen perusteella yritykselle tehtiin nykytila-analyysi, jolla pyrittiin dokumentoimaan yrityksen tietoturvakäytännöt ja määrittämään, mitä puutteita yrityksen hallinnollisissa ja teknisissä tietoturva ja -suojakäytännöissä on. Analyysin perusteella priorisoitiin kehitysprojekteja yrityksen toimintatapojen ja ohjelmistotuotteen parantamiseen. Työn tuloksena luotiin uusia ominaisuuksia työn tilaajan ohjelmistotuotteeseen. Näitä olivat rekisteröityjen suostumuksen pyyntö ja tiedotus heidän oikeuksistaan, oikeus tulla unohdetuksi ja tietojen siirto. Lisäksi tarkastettiin ja laajennettiin tilaajayrityksen tietoturvasuunnitelmaa, tietoturvaesitettä ja tietoturvaohjeistusta yrityksen henkilöstölle ja asiakkaille.Aim of this thesis was to get acquainted with EU data protection regulation 2016/679, find out which of its requirements are notable in a software company that ordered the thesis and implement a part of the changes needed. Goals of the thesis were planned together with the company. Contents of the Data protection regulation were studied and a viewed from the roles the company is in as a data controller and a data processor. Company’s clients were studied to predict what requirements they will have for the company’s main software product soon. Based on the theoretical study, a current state analysis was done to document company’s managerial and technical data protection practices and find their flaws. Based on the analysis a few development projects were formed and prioritized especially focusing on the company’s main software product. Thesis’ results were data protection documentation and guidelines for the company’s personnel and clients and software feature implementations which enable data subjects’ rights to be forgotten, to transfer their data and to gather their consent and inform them of their rights