Kinase-independent function of RIP1, critical for mature T-cell survival and proliferation.

The death receptor, Fas, triggers apoptotic death and is essential for maintaining homeostasis in the peripheral lymphoid organs. RIP1 was originally cloned when searching for Fas-binding proteins and was later shown to associate also with the signaling complex of TNFR1. Although Fas exclusively induces apoptosis, TNFR1 primarily activates the pro-survival/pro-inflammatory NF-κB pathway. Mutations in Fas lead to lymphoproliferative (lpr) diseases, and deletion of TNFR1 results in defective innate immune responses. However, the function of RIP1 in the adult lymphoid system has not been well understood, primarily owing to perinatal lethality in mice lacking the entire RIP1 protein in germ cells. This current study investigated the requirement for RIP1 in the T lineage using viable RIP1 mutant mice containing a conditional and kinase-dead RIP1 allele. Disabling the kinase activity of RIP1 had no obvious impact on the T-cell compartment. However, T-cell-specific deletion of RIP1 led to a severe T-lymphopenic condition, owing to a dramatically reduced mature T-cell pool in the periphery. Interestingly, the immature T-cell compartment in the thymus appeared intact. Further analysis showed that mature RIP1(-/-) T cells were severely defective in antigen receptor-induced proliferative responses. Moreover, the RIP1(-/-) T cells displayed greatly increased death and contained elevated caspase activities, an indication of apoptosis. In total, these results revealed a novel, kinase-independent function of RIP1, which is essential for not only promoting TCR-induced proliferative responses but also in blocking apoptosis in mature T cells

RIPK1 protects from TNF-α-mediated liver damage during hepatitis

Cell death of hepatocytes is a prominent characteristic in the pathogenesis of liver disease, while hepatolysis is a starting point of inflammation in hepatitis and loss of hepatic function. However, the precise molecular mechanisms of hepatocyte cell death, the role of the cytokines of hepatic microenvironment and the involvement of intracellular kinases, remain unclear. Tumor necrosis factor alpha (TNF-alpha) is a key cytokine involved in cell death or survival pathways and the role of RIPK1 has been associated to the TNF-alpha-dependent signaling pathway. We took advantage of two different deficient mouse lines, the RIPK1 kinase dead knock-in mice (Ripk1K45A) and the conditional knockout mice lacking RIPK1 only in liver parenchymal cells (Ripk1LPC-KO), to characterize the role of RIPK1 and TNF-alpha in hepatitis induced by concanavalin A (ConA). Our results show that RIPK1 is dispensable for liver homeostasis under steady-state conditions but in contrast, RIPK1 kinase activity contributes to caspase-independent cell death induction following ConA injection and RIPK1 also serves as a scaffold, protecting hepatocytes from massive apoptotic cell death in this model. In the Ripk1LPC-KO mice challenged with ConA, TNF-alpha triggers apoptosis, responsible for the observed severe hepatitis. Mechanism potentially involves both TNF-independent canonical NF-kappa B activation, as well as TNF-dependent, but canonical NF-kappa B-independent mechanisms. In conclusion, our results suggest that RIPK1 kinase activity is a pertinent therapeutic target to protect liver against excessive cell death in liver diseases

Astro-WISE: Chaining to the Universe

The recent explosion of recorded digital data and its processed derivatives threatens to overwhelm researchers when analysing their experimental data or when looking up data items in archives and file systems. While current hardware developments allow to acquire, process and store 100s of terabytes of data at the cost of a modern sports car, the software systems to handle these data are lagging behind. This general problem is recognized and addressed by various scientific communities, e.g., DATAGRID/EGEE federates compute and storage power over the high-energy physical community, while the astronomical community is building an Internet geared Virtual Observatory, connecting archival data. These large projects either focus on a specific distribution aspect or aim to connect many sub-communities and have a relatively long trajectory for setting standards and a common layer. Here, we report "first light" of a very different solution to the problem initiated by a smaller astronomical IT community. It provides the abstract "scientific information layer" which integrates distributed scientific analysis with distributed processing and federated archiving and publishing. By designing new abstractions and mixing in old ones, a Science Information System with fully scalable cornerstones has been achieved, transforming data systems into knowledge systems. This break-through is facilitated by the full end-to-end linking of all dependent data items, which allows full backward chaining from the observer/researcher to the experiment. Key is the notion that information is intrinsic in nature and thus is the data acquired by a scientific experiment. The new abstraction is that software systems guide the user to that intrinsic information by forcing full backward and forward chaining in the data modelling.Comment: To be published in ADASS XVI ASP Conference Series, 2006, R. Shaw, F. Hill and D. Bell, ed

Role of the caspase-1 inflammasome in Salmonella typhimurium pathogenesis

Caspase-1 is activated by a variety of stimuli after the assembly of the “inflammasome,” an activating platform made up of a complex of the NOD-LRR family of proteins. Caspase-1 is required for the secretion of proinflammatory cytokines, such as interleukin (IL)-1β and IL-18, and is involved in the control of many bacterial infections. Paradoxically, however, its absence has been reported to confer resistance to oral infection by Salmonella typhimurium. We show here that absence of caspase-1 or components of the inflammasome does not result in resistance to oral infection by S. typhimurium, but rather, leads to increased susceptibility to infection

Structure Guided Design of Potent and Selective Ponatinib-Based Hybrid Inhibitors for RIPK1

SummaryRIPK1 and RIPK3, two closely related RIPK family members, have emerged as important regulators of pathologic cell death and inflammation. In the current work, we report that the Bcr-Abl inhibitor and anti-leukemia agent ponatinib is also a first-in-class dual inhibitor of RIPK1 and RIPK3. Ponatinib potently inhibited multiple paradigms of RIPK1- and RIPK3-dependent cell death and inflammatory tumor necrosis factor alpha (TNF-α) gene transcription. We further describe design strategies that utilize the ponatinib scaffold to develop two classes of inhibitors (CS and PN series), each with greatly improved selectivity for RIPK1. In particular, we detail the development of PN10, a highly potent and selective “hybrid” RIPK1 inhibitor, capturing the best properties of two different allosteric RIPK1 inhibitors, ponatinib and necrostatin-1. Finally, we show that RIPK1 inhibitors from both classes are powerful blockers of TNF-induced injury in vivo. Altogether, these findings outline promising candidate molecules and design approaches for targeting RIPK1- and RIPK3-driven inflammatory pathologies

Directly imaging damped Lyman-alpha galaxies at z>2. I: Methodology and First Results

We present the methodology for, and the first results from, a new imaging program aimed at identifying and characterizing the host galaxies of damped Lyman-alpha absorbers (DLAs) at z>2. We target quasar sightlines with multiple optically-thick HI absorbers and use the higher-redshift system as a "blocking filter" (via its Lyman-limit absorption) to eliminate all far-ultraviolet (FUV) emission from the quasar. This allows us to directly image the rest-frame FUV continuum emission of the lower-redshift DLA, without any quasar contamination and with no bias towards large impact parameters. We introduce a formalism based on galaxy number counts and Bayesian statistics with which we quantify the probability that a candidate is the DLA host galaxy. This method will allow the identification of a bona fide sample of DLAs that are too faint to be spectroscopically confirmed. The same formalism can be adopted to the study of other quasar absorption line systems (e.g. MgII absorbers). We have applied this imaging technique to two QSO sightlines. For the z~2.69 DLA towards J073149+285449, a galaxy with impact parameter b=1.54"=11.89 kpc and implied star formation rate (SFR) of ~5 M/yr is identified as the most reliable candidate. In the case of the z~2.92 DLA towards J211444-005533, no likely host is found down to a 3-sigma SFR limit of 1.4 M/yr. Studying the HI column density as a function of the impact parameter, including 6 DLAs with known hosts from the literature, we find evidence that the observed HI distribution is more extended than what is generally predicted from numerical simulation.Comment: 22 pages, 13 figures. Accepted for publication in MNRAS. Typos correcte

Southern Cosmology Survey II: Massive Optically-Selected Clusters from 70 square degrees of the SZE Common Survey Area

We present a catalog of 105 rich and massive (M>3\times10^{14}M_{\sun}) optically-selected clusters of galaxies extracted from 70 square-degrees of public archival griz imaging from the Blanco 4-m telescope acquired over 45 nights between 2005 and 2007. We use the clusters' optically-derived properties to estimate photometric redshifts, optical luminosities, richness, and masses. We complement the optical measurements with archival XMM-Newton and ROSAT X-ray data which provide additional luminosity and mass constraints on a modest fraction of the cluster sample. Two of our clusters show clear evidence for central lensing arcs; one of these has a spectacular large-diameter, nearly-complete Einstein Ring surrounding the brightest cluster galaxy. A strong motivation for this study is to identify the massive clusters that are expected to display prominent signals from the Sunyaev-Zeldovich Effect (SZE) and therefore be detected in the wide-area mm-band surveys being conducted by both the Atacama Cosmology Telescope and the South Pole Telescope. The optical sample presented here will be useful for verifying new SZE cluster candidates from these surveys, for testing the cluster selection function, and for stacking analyzes of the SZE data.Comment: 13 pages, 7 Figures. Accepted for publication to ApJSS. Full resolution plots and additional material available at http://peumo.rutgers.edu/~felipe/e-prints

Respiratory Syncytial Virus infection promotes necroptosis and HMGB1 release by airway epithelial cells

Rationale: Respiratory syncytial virus (RSV) bronchiolitis causes significant infant mortality. Bronchiolitis is characterized by airway epithelial cell (AEC) death; however, the mode of death remains unknown. Objectives: To determine whether necroptosis contributes to RSV b r onchiolitis pathogenesis via HMGB1 (high mobility group box 1) release. Methods: Nasopharyngeal samples were collected from children presenting to the hospital with acute respiratory infection. Primary human AECs and neonatal mice were inoculated with RSV and murine Pneumovirus, respectively. Necroptosis was determined via viability assays and immunohistochemistry for RIPK1 (receptor-interacting protein kinase-1), MLKL (mixed lineage kinase domain-like pseudokinase) protein, and caspase-3. Necroptosis was blocked using pharmacological inhibitors and RIPK1 kinase-dead knockin mice. Measurements and Main Results: HMGB1 levels were elevated in nasopharyngeal samples of children with acute RSV infection. RSV-induced epithelial cell death was associated with increased phosphorylated RIPK1 and phosphorylated MLKL but not active caspase-3 expression. Inhibition of RIPK1 or MLKL attenuated RSV-induced HMGBI translocation and release, and lowered viral load. MLKL inhibition increased active caspase-3 expression in a caspase-8/9-dependent manner. In susceptible mice, Pneumovirus infection upregulated RIPK1 and MLKL expression in the airway epithelium at 8 to 10 days after infection, coinciding with AEC sloughing, HMGB1 release, and neutrophilic inflammation. Genetic or pharmacological inhibition of RIPK1 or MLKL attenuated these pathologies, lowered viral load, and prevented type 2 inflammation and airway remodeling. Necroptosis inhibition in early life ameliorated asthma progression induced by viral or allergen challenge in later life. Conclusions: Pneumovirus infection induces AEC necroptosis. Inhibition of necroptosis may be a viable strategy to limit the severity of viral bronchiolitis and break its nexus with asthma