Analysis of the role of Pellino2 in the Activation of the NLRP3 Inflammasome.

The innate immune system is the first line of defence against invading pathogens and is responsible for both initial pathogen recognition and mounting an effective immunological response. The recognition of conserved pathogen-associated molecular patterns can be carried out by a number of families of innate immune receptors, including Toll-like receptors. Toll-like receptors can respond to a diverse range of microbial signals and elicit a targeted immune response against the invading pathogen. A key regulator of inflammation is IL-1β, which can be secreted from activated cells and mediate a strong inflammatory response. The regulation of its release is a tightly controlled signalling event, as its dysregulation has been linked with the pathogenesis of a variety of diseases including Alzheimer’s disease and rheumatoid arthritis. The signalling complex which controls IL-1β release is known as the inflammasome, which requires two signals to facilitate its activation. The work in this thesis demonstrates a role for the E3 ubiquitin ligase Pellino2 in the specific activation of the NLRP3 inflammasome. Using a newly generated Pellino2-deficient mouse, it was determined that Pellino2 played no role during TLR induced NF-κB or MAPK signalling. However, Peli2-/- mice were more resistant to the lethal effects of LPS induced septic shock and this correlated with reduced serum IL-1β levels. Pellino2-deficient mice were shown to exhibit diminished NLRP3 inflammasome activation, correlating with reduced levels of NLRP3 ubiquitination and thus, lower levels of IL-1β secretion. While Pellino2 does not directly mediate the ubiquitination of NLRP3, it was observed that Pellino2-deficiency promoted the increased interaction of NLRP3 with hyperphosphorylated IRAK1. The data presented in this thesis defines for the first time a physiological role for Pellino2 in innate immune signalling with a particular function in mediating activation of the NLRP3 inflammasome

The influence of motivation and comfort-level on learning to program

This paper documents a study, carried out in the academic year 2004-2005, on the role of motivation and comfort-level in a first year object-oriented programming module. The study found that intrinsic motivation had a strong correlation with programming performance as did self-efficacy for learning and performance, r=0.512, p < 0.01 and r=0.567, p < 0.01 respectively. Aspects of comfort level were found to have significant correlations with performance with an instrument on programming-esteem rendering the most interesting results. A regression model based upon these factors was able to account for 60% of the variance in programming performance results

The influence of motivation and comfort-level on learning to program

This paper documents a study, carried out in the academic year 2004-2005, on the role of motivation and comfort-level in a first year object-oriented programming module. The study found that intrinsic motivation had a strong correlation with programming performance as did self-efficacy for learning and performance, r=0.512, p < 0.01 and r=0.567, p < 0.01 respectively. Aspects of comfort level were found to have significant correlations with performance with an instrument on programming-esteem rendering the most interesting results. A regression model based upon these factors was able to account for 60% of the variance in programming performance results

Towards Automatic Blotch Detection for Film Restoration by Comparison of Spatio-Temporal Neighbours

In this paper, a new method of blotch detection for digitised film sequences is proposed. Due to the aging of film stocks, their poor storage and/or repeated viewing, it is estimated that approximately 50% of all films produced prior to 1950 have either been destroyed or rendered unwatchable [1,2]. To prevent their complete destruction, original film reels must be scanned into digital format; however, any defects such as blotches will be retained. By combining a variation of a linear time, contour tracing technique with a simple temporal nearest neighbour algorithm, a preliminary detection system has been created. Using component labelling of dirt and sparkle the overall performance of the completed system, in terms of time and accuracy, will compare favourably to traditional motion compensated detection methods. This small study (based on 13 film sequences) represents a significant first step towards automatic blotch detection

Effect of Hydrocortisone on Mortality and Organ Support in Patients With Severe COVID-19: The REMAP-CAP COVID-19 Corticosteroid Domain Randomized Clinical Trial.

Importance: Evidence regarding corticosteroid use for severe coronavirus disease 2019 (COVID-19) is limited. Objective: To determine whether hydrocortisone improves outcome for patients with severe COVID-19. Design, Setting, and Participants: An ongoing adaptive platform trial testing multiple interventions within multiple therapeutic domains, for example, antiviral agents, corticosteroids, or immunoglobulin. Between March 9 and June 17, 2020, 614 adult patients with suspected or confirmed COVID-19 were enrolled and randomized within at least 1 domain following admission to an intensive care unit (ICU) for respiratory or cardiovascular organ support at 121 sites in 8 countries. Of these, 403 were randomized to open-label interventions within the corticosteroid domain. The domain was halted after results from another trial were released. Follow-up ended August 12, 2020. Interventions: The corticosteroid domain randomized participants to a fixed 7-day course of intravenous hydrocortisone (50 mg or 100 mg every 6 hours) (n = 143), a shock-dependent course (50 mg every 6 hours when shock was clinically evident) (n = 152), or no hydrocortisone (n = 108). Main Outcomes and Measures: The primary end point was organ support-free days (days alive and free of ICU-based respiratory or cardiovascular support) within 21 days, where patients who died were assigned -1 day. The primary analysis was a bayesian cumulative logistic model that included all patients enrolled with severe COVID-19, adjusting for age, sex, site, region, time, assignment to interventions within other domains, and domain and intervention eligibility. Superiority was defined as the posterior probability of an odds ratio greater than 1 (threshold for trial conclusion of superiority >99%). Results: After excluding 19 participants who withdrew consent, there were 384 patients (mean age, 60 years; 29% female) randomized to the fixed-dose (n = 137), shock-dependent (n = 146), and no (n = 101) hydrocortisone groups; 379 (99%) completed the study and were included in the analysis. The mean age for the 3 groups ranged between 59.5 and 60.4 years; most patients were male (range, 70.6%-71.5%); mean body mass index ranged between 29.7 and 30.9; and patients receiving mechanical ventilation ranged between 50.0% and 63.5%. For the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively, the median organ support-free days were 0 (IQR, -1 to 15), 0 (IQR, -1 to 13), and 0 (-1 to 11) days (composed of 30%, 26%, and 33% mortality rates and 11.5, 9.5, and 6 median organ support-free days among survivors). The median adjusted odds ratio and bayesian probability of superiority were 1.43 (95% credible interval, 0.91-2.27) and 93% for fixed-dose hydrocortisone, respectively, and were 1.22 (95% credible interval, 0.76-1.94) and 80% for shock-dependent hydrocortisone compared with no hydrocortisone. Serious adverse events were reported in 4 (3%), 5 (3%), and 1 (1%) patients in the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively. Conclusions and Relevance: Among patients with severe COVID-19, treatment with a 7-day fixed-dose course of hydrocortisone or shock-dependent dosing of hydrocortisone, compared with no hydrocortisone, resulted in 93% and 80% probabilities of superiority with regard to the odds of improvement in organ support-free days within 21 days. However, the trial was stopped early and no treatment strategy met prespecified criteria for statistical superiority, precluding definitive conclusions. Trial Registration: ClinicalTrials.gov Identifier: NCT02735707

Circulating microRNAs in sera correlate with soluble biomarkers of immune activation but do not predict mortality in ART treated individuals with HIV-1 infection: A case control study

Introduction: The use of anti-retroviral therapy (ART) has dramatically reduced HIV-1 associated morbidity and mortality. However, HIV-1 infected individuals have increased rates of morbidity and mortality compared to the non-HIV-1 infected population and this appears to be related to end-organ diseases collectively referred to as Serious Non-AIDS Events (SNAEs). Circulating miRNAs are reported as promising biomarkers for a number of human disease conditions including those that constitute SNAEs. Our study sought to investigate the potential of selected miRNAs in predicting mortality in HIV-1 infected ART treated individuals. Materials and Methods: A set of miRNAs was chosen based on published associations with human disease conditions that constitute SNAEs. This case: control study compared 126 cases (individuals who died whilst on therapy), and 247 matched controls (individuals who remained alive). Cases and controls were ART treated participants of two pivotal HIV-1 trials. The relative abundance of each miRNA in serum was measured, by RTqPCR. Associations with mortality (all-cause, cardiovascular and malignancy) were assessed by logistic regression analysis. Correlations between miRNAs and CD4+ T cell count, hs-CRP, IL-6 and D-dimer were also assessed. Results: None of the selected miRNAs was associated with all-cause, cardiovascular or malignancy mortality. The levels of three miRNAs (miRs -21, -122 and -200a) correlated with IL-6 while miR-21 also correlated with D-dimer. Additionally, the abundance of miRs -31, -150 and -223, correlated with baseline CD4+ T cell count while the same three miRNAs plus miR- 145 correlated with nadir CD4+ T cell count. Discussion: No associations with mortality were found with any circulating miRNA studied. These results cast doubt onto the effectiveness of circulating miRNA as early predictors of mortality or the major underlying diseases that contribute to mortality in participants treated for HIV-1 infection

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Development and Validation of a Risk Score for Chronic Kidney Disease in HIV Infection Using Prospective Cohort Data from the D:A:D Study

Ristola M. on työryhmien DAD Study Grp ; Royal Free Hosp Clin Cohort ; INSIGHT Study Grp ; SMART Study Grp ; ESPRIT Study Grp jäsen.Background Chronic kidney disease (CKD) is a major health issue for HIV-positive individuals, associated with increased morbidity and mortality. Development and implementation of a risk score model for CKD would allow comparison of the risks and benefits of adding potentially nephrotoxic antiretrovirals to a treatment regimen and would identify those at greatest risk of CKD. The aims of this study were to develop a simple, externally validated, and widely applicable long-term risk score model for CKD in HIV-positive individuals that can guide decision making in clinical practice. Methods and Findings A total of 17,954 HIV-positive individuals from the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D) study with >= 3 estimated glomerular filtration rate (eGFR) values after 1 January 2004 were included. Baseline was defined as the first eGFR > 60 ml/min/1.73 m2 after 1 January 2004; individuals with exposure to tenofovir, atazanavir, atazanavir/ritonavir, lopinavir/ritonavir, other boosted protease inhibitors before baseline were excluded. CKD was defined as confirmed (>3 mo apart) eGFR In the D:A:D study, 641 individuals developed CKD during 103,185 person-years of follow-up (PYFU; incidence 6.2/1,000 PYFU, 95% CI 5.7-6.7; median follow-up 6.1 y, range 0.3-9.1 y). Older age, intravenous drug use, hepatitis C coinfection, lower baseline eGFR, female gender, lower CD4 count nadir, hypertension, diabetes, and cardiovascular disease (CVD) predicted CKD. The adjusted incidence rate ratios of these nine categorical variables were scaled and summed to create the risk score. The median risk score at baseline was -2 (interquartile range -4 to 2). There was a 1: 393 chance of developing CKD in the next 5 y in the low risk group (risk score = 5, 505 events), respectively. Number needed to harm (NNTH) at 5 y when starting unboosted atazanavir or lopinavir/ritonavir among those with a low risk score was 1,702 (95% CI 1,166-3,367); NNTH was 202 (95% CI 159-278) and 21 (95% CI 19-23), respectively, for those with a medium and high risk score. NNTH was 739 (95% CI 506-1462), 88 (95% CI 69-121), and 9 (95% CI 8-10) for those with a low, medium, and high risk score, respectively, starting tenofovir, atazanavir/ritonavir, or another boosted protease inhibitor. The Royal Free Hospital Clinic Cohort included 2,548 individuals, of whom 94 individuals developed CKD (3.7%) during 18,376 PYFU (median follow-up 7.4 y, range 0.3-12.7 y). Of 2,013 individuals included from the SMART/ESPRIT control arms, 32 individuals developed CKD (1.6%) during 8,452 PYFU (median follow-up 4.1 y, range 0.6-8.1 y). External validation showed that the risk score predicted well in these cohorts. Limitations of this study included limited data on race and no information on proteinuria. Conclusions Both traditional and HIV-related risk factors were predictive of CKD. These factors were used to develop a risk score for CKD in HIV infection, externally validated, that has direct clinical relevance for patients and clinicians to weigh the benefits of certain antiretrovirals against the risk of CKD and to identify those at greatest risk of CKD.Peer reviewe

Analysis of the role of Pellino2 in the Activation of the NLRP3 Inflammasome.

The innate immune system is the first line of defence against invading pathogens and is responsible for both initial pathogen recognition and mounting an effective immunological response. The recognition of conserved pathogen-associated molecular patterns can be carried out by a number of families of innate immune receptors, including Toll-like receptors. Toll-like receptors can respond to a diverse range of microbial signals and elicit a targeted immune response against the invading pathogen. A key regulator of inflammation is IL-1β, which can be secreted from activated cells and mediate a strong inflammatory response. The regulation of its release is a tightly controlled signalling event, as its dysregulation has been linked with the pathogenesis of a variety of diseases including Alzheimer’s disease and rheumatoid arthritis. The signalling complex which controls IL-1β release is known as the inflammasome, which requires two signals to facilitate its activation. The work in this thesis demonstrates a role for the E3 ubiquitin ligase Pellino2 in the specific activation of the NLRP3 inflammasome. Using a newly generated Pellino2-deficient mouse, it was determined that Pellino2 played no role during TLR induced NF-κB or MAPK signalling. However, Peli2-/- mice were more resistant to the lethal effects of LPS induced septic shock and this correlated with reduced serum IL-1β levels. Pellino2-deficient mice were shown to exhibit diminished NLRP3 inflammasome activation, correlating with reduced levels of NLRP3 ubiquitination and thus, lower levels of IL-1β secretion. While Pellino2 does not directly mediate the ubiquitination of NLRP3, it was observed that Pellino2-deficiency promoted the increased interaction of NLRP3 with hyperphosphorylated IRAK1. The data presented in this thesis defines for the first time a physiological role for Pellino2 in innate immune signalling with a particular function in mediating activation of the NLRP3 inflammasome