Differential Binding of NLRP3 to non-oxidized and Ox-mtDNA mediates NLRP3 Inflammasome Activation.

The NLRP3 inflammasome is a key mediator of the innate immune response to sterile tissue injury and is involved in many chronic and acute diseases. Physically and chemically diverse agents activate the NLRP3 inflammasome. Here, we show that NLRP3 binds non-oxidized and Ox-mtDNA differentially, with a half maximum inhibitory concentration (IC50) for non-oxidized and Ox-mtDNA of 4 nM and 247.2 nM, respectively. The NLRP3 Neonatal-Onset Multisystem Inflammatory Disease (NOMIDFCAS) gain of function mutant could bind non-oxidized mtDNA but had higher affinity for Ox-mtDNA compared to WT with an IC50 of 8.1 nM. NLRP3 lacking the pyrin domain can bind both oxidized and non-oxidized mtDNA. Isolated pyrin domain prefers Ox-mtDNA. The NLRP3 pyrin domain shares a protein fold with DNA glycosylases and generate a model for DNA binding based on the structure and sequence alignment to Clostridium acetobutylicum and human OGG1, an inhibitor of Ox-mtDNA generation, 8-oxoguanine DNA glycosylases. We provide a new model for how NLRP3 interacts with Ox-mtDNA supported by DNA binding in the presence of a monoclonal antibody against the pyrin domain. These results give new insights into the mechanism of inflammasome assembly, and into the function of reactive oxygen species in establishing a robust immune response.

Cryo-EM for Small Molecules.

In recent years, protein structure analysis using cryo-electron microscopy(cryo-EM) has expanded and improved. In this review, we discuss many recent improvements to the field, the problems those improvements hope to solve, and some of the still unanswered questions. Most notably, this review will discuss improvements in resolving small or fragmented protein structures, as well as methods to improve the signal-to-noise ratio of the data by increasing image contrast using carbon-based systems. We will also describe how, in the last 5 years, methodological improvements have allowed for better 3D image resolution by capturing a continuum of 3D images. We will provide examples of these methods in practice and discuss how these improved methods may be used in small-molecule drug discovery and development. © 2022 Wiley Periodicals LLC.

Improved sanitation and income are associated with decreased rates of hospitalization for diarrhoea in Brazilian infants.

Diarrhoeal diseases remain a major cause of morbidity and mortality in Brazilian children. However, from 1992 to 2001 there was a significant decline in hospitalizations for acute diarrhoea in children below 1 year of age in Brazil. A significant improvement in child health was also observed in the state of Rio Grande do Norte (RN), with a decrease in child mortality from 70 to 40 deaths per 1000. Using distributed lag analysis we analysed a number of factors possibly connected with decreased hospitalization in RN and found that hospitalization was correlated up to lag 3 with poverty (P<0.001) and inflation (P<0.001). Improvements in public health infrastructure such as better waste collection, presence of city water supply and increased sanitation, socio-economic variables such as education and literacy, and increased investment in health services were all important in reducing severe early childhood diarrhoeas and thus directly associated with the decrease in hospitalization. We also observed a positive seasonal correlation between rainfall and hospitalizations with an increased in rainfall impacting positively on hospitalization in all lags. The data suggests that increased buying power and reductions in poverty played a crucial role in reducing hospitalizations for acute diarrhoea in infants in RN.

Genetic predisposition to self-curing infection with the protozoan Leishmania chagasi: a genomewide scan.

The protozoan Leishmania chagasi can cause disseminated, fatal visceral leishmaniasis (VL) or asymptomatic infection in humans. We hypothesized that host genetic factors contribute to this variable response to infection. A family study was performed in neighborhoods of endemicity for L. chagasi near Natal in northeastern Brazil. Study subjects were assessed for the presence of VL or asymptomatic infection, which was defined by a positive delayed-type hypersensitivity (DTH) skin test response to Leishmania antigen without disease symptoms. A genomewide panel of 385 autosomal microsatellite markers in 1254 subjects from 191 families was analyzed to identify regions of linkage. Regions with potential linkage to the DTH response on chromosomes 15 and 19, as well as a novel region on chromosome 9 with potential linkage to VL, were identified. Understanding the genetic factors that determine whether an individual will develop symptomatic or asymptomatic infection with L. chagasi may identify proteins essential for immune protection against this parasitic disease and reveal strategies for immunotherapy or prevention.