Neonatal plasmacytoid dendritic cells at the interface of innate and adaptive immunity = Neonatale plasmazytoide dendritische Zellen - an der Schnittstelle zwischen angeborener und adaptiver Immunität

Human cord blood plasmacytoid dendritic cells (PDC) react to stimulation with CPG A and CPG B with an increase in cell surface activation and maturation markers and cytokine production, similar to adult PDC. Intracellular phosphorylation in neonatal PDC did not benefit from CPG stimulation, in contrast to adult PDC. Cord blood PDC primed with CPG A, CPG B and CD40L do not promote division of autologous T cells contrary to adult PDC. Priming of neonatal PDC with CPG A or CPG B does not induce a clear bias in T helper cell response towards Th1 or Th2 while adult PDC trend towards a Th2 response.

The interaction of Leishmania major parasites with human myeloid cells and its consequence for adaptive immunity

In der vorliegenden Arbeit fokussierten wir uns auf drei verschiedene Aspekte der Leishmanien-Infektion. Wir charakterisierten den Prozess des Zelltods „Apoptose“ bei Parasiten (1), untersuchten die Eignung von Makrophagen und dendritischen Zellen als Wirtszelle für die Entwicklung der Parasiten (2) und analysierten die Konsequenzen der Infektion für die Entstehung einer adaptiven Immunantwort im humanen System. Von zentraler Bedeutung für dieses Projekt war die Hypothese, dass apoptotische Leishmanien den Autophagie-Mechanismus ihrer Wirtszellen ausnutzen, um eine T-Zell-vermittelte Abtötung der Parasiten zu vermindern.rnWir definierten eine apoptotische Leishmanien-Population, welche durch eine rundliche Morphologie und die Expression von Phosphatidylserin auf der Parasitenoberfläche charakterisiert war. Die apoptotischen Parasiten befanden sich zudem in der SubG1-Phase und wiesen weniger und fragmentierte DNA auf, welche durch TUNEL-Assay nachgewiesen werden konnte. Bei der Interaktion der Parasiten mit humanen Makrophagen und dendritischen Zellen zeigte sich, dass die anti-inflammatorischen Makrophagen anfälliger für Infektionen waren als die pro-inflammatorischen Makrophagen oder die dendritischen Zellen. Interessanterweise wurde in den dendritischen Zellen jedoch die effektivste Umwandlung zur krankheitsauslösenden, amastigoten Lebensform beobachtet. Da sowohl Makrophagen als auch dendritische Zellen zu den antigenpräsentierenden Zellen gehören, könnte dies zur Aktivierung der T-Zellen des adaptiven Immunsystems führen. Tatsächlich konnte während der Leishmanien-Infektion die Proliferation von T-Zellen beobachtet werden. Dabei stellten wir fest, dass es sich bei den proliferierenden T-Zellen um CD3+CD4+ T-Zellen handelte, welche sich überraschenderweise als Leishmanien-spezifische CD45RO+ T-Gedächtniszellen herausstellten. Dies war unerwartet, da ein vorheriger Kontakt der Spender mit Leishmanien als unwahrscheinlich gilt. In Gegenwart von apoptotischen Parasiten konnte eine signifikant schwächere T-Zell-Proliferation in Makrophagen, jedoch nicht in dendritischen Zellen beobachtet werden. Da sich die T-Zell-Proliferation negativ auf das Überleben der Parasiten auswirkt, konnten die niedrigsten Überlebensraten in dendritischen Zellen vorgefunden werden. Innerhalb der Zellen befanden sich die Parasiten in beiden Zelltypen im Phagosom, welches allerdings nur in Makrophagen den Autophagie-Marker LC3 aufwies. Chemische Induktion von Autophagie führte, ebenso wie die Anwesenheit von apoptotischen Parasiten, zu einer stark reduzierten T-Zell-Proliferation und dementsprechend zu einem höheren Überleben der Parasiten.rnZusammenfassend lässt sich aus unseren Daten schließen, dass Apoptose in Einzellern vorkommt. Während der Infektion können sowohl Makrophagen, als auch dendritische Zellen mit Leishmanien infiziert und das adaptive Immunsystem aktivert werden. Die eingeleitete T-Zell-Proliferation nach Infektion von Makrophagen ist in Gegenwart von apoptotischen Parasiten reduziert, weshalb sie im Vergleich zu dendritischen Zellen die geeigneteren Wirtszellen für Leishmanien darstellen. Dafür missbrauchen die Parasiten den Autophagie-Mechanismus der Makrophagen als Fluchtstrategie um das adaptive Immunsystem zu umgehen und somit das Überleben der Gesamtpopulation zu sichern. Diese Ergebnisse erklären den Vorteil von Apoptose in Einzellern und verdeutlichen, dass der Autophagie-Mechanismus als potentielles therapeutisches Ziel für die Behandlung von Leishmaniose dienen kann.rn

Dynamic Adaptive Streaming over Http: implementazione e analisi sperimentale di algoritmi di rate adaptation su client Android

Il lavoro svolto in questa tesi consiste nell'implementare un'applicazione Android per lo streaming video, conforme allo standard MPEG-DASH. L'obiettivo è quello di fornire un valido strumento al fine di eseguire delle analisi sperimentali su algoritmi particolari, detti di rate adaptation. MPEG-Dynamic Adaptive Streaming over Http è uno standard emergente ed è considerato da molti il futuro dello streaming multimediale. Questa tecnologia consente di auto-regolare la qualità del video in base alle condizioni della rete, la capacità del dispositivo o le preferenze dell'utente. Inoltre, essendo uno standard, permette di rendere interoperabili i server e i device dei vari fornitori di contenuti multimediali. Nei primi capitoli introduttivi verrà presentato lo standard e i lavori correlati, successivamente verrà descritta la mia proposta applicativa: DashPlayer. In conclusione, verrà compiuta una valutazione sperimentali sugli algoritmi sopracitati che costituiscono la parte logico-funzionale dell'applicazione.

Evaluation of new HTTP Adaptive Streaming algorithms: the clients’ and network’s perspectives

Lo streaming è una tecnica per trasferire contenuti multimediali sulla rete globale, utilizzato per esempio da servizi come YouTube e Netflix; dopo una breve attesa, durante la quale un buffer di sicurezza viene riempito, l'utente può usufruire del contenuto richiesto. Cisco e Sandvine, che con cadenza regolare pubblicano bollettini sullo stato di Internet, affermano che lo streaming video ha, e avrà sempre di più, un grande impatto sulla rete globale. Il buon design delle applicazioni di streaming riveste quindi un ruolo importante, sia per la soddisfazione degli utenti che per la stabilità dell'infrastruttura. HTTP Adaptive Streaming indica una famiglia di implementazioni volta a offrire la migliore qualità video possibile (in termini di bit rate) in funzione della bontà della connessione Internet dell'utente finale: il riproduttore multimediale può cambiare in ogni momento il bit rate, scegliendolo in un insieme predefinito, adattandosi alle condizioni della rete. Per ricavare informazioni sullo stato della connettività, due famiglie di metodi sono possibili: misurare la velocità di scaricamento dei precedenti trasferimenti (approccio rate-based), oppure, come recentemente proposto da Netflix, utilizzare l'occupazione del buffer come dato principale (buffer-based). In questo lavoro analizziamo algoritmi di adattamento delle due famiglie, con l'obiettivo di confrontarli su metriche riguardanti la soddisfazione degli utenti, l'utilizzo della rete e la competizione su un collo di bottiglia. I risultati dei nostri test non definiscono un chiaro vincitore, riconoscendo comunque la bontà della nuova proposta, ma evidenziando al contrario che gli algoritmi buffer-based non sempre riescono ad allocare in modo imparziale le risorse di rete.

Implementazione e analisi di un’applicazione Android per il Dynamic Adaptive Streaming cooperativo tramite Http e WiFi-Direct

Questo lavoro ha lo scopo di presentare l’implementazione e la valutazione di un’applicazione Android che permetta la riproduzione di uno streaming auto-adattante conforme allo standard DASH sfruttando le funzionalità offerte dal player ”ExoPLayer” a cui viene aggiunta la funzionalità di caching e condivisione dei relativi segmenti tramite WiFi-Direct. Questo al fine di raggiungere diversi obiettivi come la riduzione dell’utilizzo di reti mobili, l’aumento della qualità e/o una riproduzione più fluida. Si è inoltre sviluppato un simulatore in C++ che permette di valutare il comportamento dell’applicazione, l’algoritmo usato per la scelta dei segmenti, e i vantaggi correlati.

Tecniche Cloud Avanzate per Adaptive Hadoop Load Balancing tramite Sahara

Ogni giorno vengono generati grandi moli di dati attraverso sorgenti diverse. Questi dati, chiamati Big Data, sono attualmente oggetto di forte interesse nel settore IT (Information Technology). I processi digitalizzati, le interazioni sui social media, i sensori ed i sistemi mobili, che utilizziamo quotidianamente, sono solo un piccolo sottoinsieme di tutte le fonti che contribuiscono alla produzione di questi dati. Per poter analizzare ed estrarre informazioni da questi grandi volumi di dati, tante sono le tecnologie che sono state sviluppate. Molte di queste sfruttano approcci distribuiti e paralleli. Una delle tecnologie che ha avuto maggior successo nel processamento dei Big Data, e Apache Hadoop. Il Cloud Computing, in particolare le soluzioni che seguono il modello IaaS (Infrastructure as a Service), forniscono un valido strumento all'approvvigionamento di risorse in maniera semplice e veloce. Per questo motivo, in questa proposta, viene utilizzato OpenStack come piattaforma IaaS. Grazie all'integrazione delle tecnologie OpenStack e Hadoop, attraverso Sahara, si riesce a sfruttare le potenzialita offerte da un ambiente cloud per migliorare le prestazioni dell'elaborazione distribuita e parallela. Lo scopo di questo lavoro e ottenere una miglior distribuzione delle risorse utilizzate nel sistema cloud con obiettivi di load balancing. Per raggiungere questi obiettivi, si sono rese necessarie modifiche sia al framework Hadoop che al progetto Sahara.

Blood glucose and prognosis in children with presumed severe malaria: is there a threshold for 'hypoglycaemia'?

OBJECTIVES: Hypoglycaemia (glucose <2.2 mmol/l) is a defining feature of severe malaria, but the significance of other levels of blood glucose has not previously been studied in children with severe malaria. METHODS: A prospective study of 437 consecutive children with presumed severe malaria was conducted in Mali. We defined hypoglycaemia as <2.2 mmol/l, low glycaemia as 2.2-4.4 mmol/l and hyperglycaemia as >8.3 mmol/l. Associations between glycaemia and case fatality were analysed for 418 children using logistic regression models and a receiver operator curve (ROC). RESULTS: There was a significant difference between blood glucose levels in children who died (median 4.6 mmol/l) and survivors (median 7.6 mmol/l, P < 0.001). Case fatality declined from 61.5% of the hypoglycaemic children to 46.2% of those with low glycaemia, 13.4% of those with normal glycaemia and 7.6% of those with hyperglycaemia (P < 0.001). Logistic regression showed an adjusted odds ratio (AOR) of 0.75 (0.64-0.88) for case fatality per 1 mmol/l increase in baseline blood glucose. Compared to a normal blood glucose, hypoglycaemia and low glycaemia both significantly increased the odds of death (AOR 11.87, 2.10-67.00; and 5.21, 1.86-14.63, respectively), whereas hyperglycaemia reduced the odds of death (AOR 0.34, 0.13-0.91). The ROC [area under the curve at 0.753 (95% CI 0.684-0.820)] indicated that glycaemia had a moderate predictive value for death and identified an optimal threshold at glycaemia <6.1 mmol/l, (sensitivity 64.5% and specificity 75.1%). CONCLUSIONS: If there is a threshold of blood glucose which defines a worse prognosis, it is at a higher level than the current definition of 2.2 mmol/l.

Urinary metabolomics in Fxr-null mice reveals activated adaptive metabolic pathways upon bile acid challenge

Farnesoid X receptor (FXR) is a nuclear receptor that regulates genes involved in synthesis, metabolism, and transport of bile acids and thus plays a major role in maintaining bile acid homeostasis. In this study, metabolomic responses were investigated in urine of wild-type and Fxr-null mice fed cholic acid, an FXR ligand, using ultra-performance liquid chromatography (UPLC) coupled with electrospray time-of-flight mass spectrometry (TOFMS). Multivariate data analysis between wild-type and Fxr-null mice on a cholic acid diet revealed that the most increased ions were metabolites of p-cresol (4-methylphenol), corticosterone, and cholic acid in Fxr-null mice. The structural identities of the above metabolites were confirmed by chemical synthesis and by comparing retention time (RT) and/or tandem mass fragmentation patterns of the urinary metabolites with the authentic standards. Tauro-3alpha,6,7alpha,12alpha-tetrol (3alpha,6,7alpha,12alpha-tetrahydroxy-5beta-cholestan-26-oyltaurine), one of the most increased metabolites in Fxr-null mice on a CA diet, is a marker for efficient hydroxylation of toxic bile acids possibly through induction of Cyp3a11. A cholestatic model induced by lithocholic acid revealed that enhanced expression of Cyp3a11 is the major defense mechanism to detoxify cholestatic bile acids in Fxr-null mice. These results will be useful for identification of biomarkers for cholestasis and for determination of adaptive molecular mechanisms in cholestasis.

Mitochondrial tRNA(Leu(UUR)) mutation m.3302A > G presenting as childhood-onset severe myopathy: threshold determination through segregation study

Mitochondrial tRNA(Leu(UUR)) mutation m.3302A > G is associated with respiratory chain complex I deficiency and has been described as a rare cause of mostly adult-onset slowly progressive myopathy. Five families with 11 patients have been described so far; 5 of them died young due to cardiorespiratory failure. Here, we report on a segregation study in a family with an index patient who already presented at the age of 18 months with proximal muscular hypotonia, abnormal fatigability, and lactic acidosis. This early-onset myopathy was rapidly progressive. At 8 years, the patient is wheel-chair bound, requires nocturnal assisted ventilation, and suffers from recurrent respiratory infections. Severe complex I deficiency and nearly homoplasmy for m.3302A > G were found in muscle. We collected blood, hair, buccal swabs and muscle biopsies from asymptomatic adults in this pedigree and determined heteroplasmy levels in these tissues as well as OXPHOS activities in muscle. All participating asymptomatic adults had normal OXPHOS activities. In contrast to earlier reports, we found surprisingly little variation of heteroplasmy levels in different tissues of the same individual. Up to 45% mutation load in muscle and up to 38% mutation load in other tissues were found in non-affected adults. The phenotypic spectrum of tRNA(Leu(UUR)) m.3302A > G mutation seems to be wider than previously described. A threshold of more than 45% heteroplasmy in muscle seems to be necessary to alter complex I activity leading to clinical manifestation. The presented data may be helpful for prognostic considerations and counseling in affected families.

Low-tube-voltage, high-tube-current multidetector abdominal CT: improved image quality and decreased radiation dose with adaptive statistical iterative reconstruction algorithm--initial clinical experience

To investigate whether an adaptive statistical iterative reconstruction (ASIR) algorithm improves the image quality at low-tube-voltage (80-kVp), high-tube-current (675-mA) multidetector abdominal computed tomography (CT) during the late hepatic arterial phase.

Properties of low-threshold motor axons in the human median nerve

This study investigated the excitability and accommodative properties of low-threshold human motor axons to test whether these motor axons have greater expression of the persistent Na(+) conductance, I(NaP). Computer-controlled threshold tracking was used to study 22 single motor units and the data were compared with compound motor potentials of various amplitudes recorded in the same experimental session. Detailed comparisons were made between the single units and compound potentials that were 40% or 5% of maximal amplitude, the former because this is the compound potential size used in most threshold tracking studies of axonal excitability, the latter because this is the compound potential most likely to be composed entirely of motor axons with low thresholds to electrical recruitment. Measurements were made of the strength-duration relationship, threshold electrotonus, current-voltage relationship, recovery cycle and latent addition. The findings did not support a difference in I(NaP). Instead they pointed to greater activity of the hyperpolarization-activated inwardly rectifying current (I(h)) as the basis for low threshold to electrical recruitment in human motor axons. Computer modelling confirmed this finding, with a doubling of the hyperpolarization-activated conductance proving the best single parameter adjustment to fit the experimental data. We suggest that the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel(s) expressed on human motor axons may be active at rest and contribute to resting membrane potential.