Xanthine oxidoreductase regulates macrophage IL1β secretion upon NLRP3 inflammasome activation.

Activation of the NLRP3 inflammasome by microbial ligands or tissue damage requires intracellular generation of reactive oxygen species (ROS). We present evidence that macrophage secretion of IL1β upon stimulation with ATP, crystals or LPS is mediated by a rapid increase in the activity of xanthine oxidase (XO), the oxidized form of xanthine dehydrogenase, resulting in the formation of uric acid as well as ROS. We show that XO-derived ROS, but not uric acid, is the trigger for IL1β release and that XO blockade results in impaired IL1β and caspase1 secretion. XO is localized to both cytoplasmic and mitochondrial compartments and acts upstream to the PI3K-AKT signalling pathway that results in mitochondrial ROS generation. This pathway represents a mechanism for regulating NLRP3 inflammasome activation that may have therapeutic implications in inflammatory diseases

STING gain-of-function disrupts lymph node organogenesis and innate lymphoid cell development in mice

STING gain-of-function causes autoimmunity and immunodeficiency in mice and STING-associated vasculopathy with onset in infancy (SAVI) in humans. Here, we report that STING gain-of-function in mice prevents development of lymph nodes and Peyer\u27s patches. We show that the absence of secondary lymphoid organs is associated with diminished numbers of innate lymphoid cells (ILCs), including lymphoid tissue inducer (LTi) cells. Although wild-type (WT) α4β

New measures to capture end of life concerns in Huntington disease: Meaning and Purpose and Concern with Death and Dying from HDQLIFE (a patient-reported outcomes measurement system).

PURPOSE: Huntington disease (HD) is an incurable terminal disease. Thus, end of life (EOL) concerns are common in these individuals. A quantitative measure of EOL concerns in HD would enable a better understanding of how these concerns impact health-related quality of life. Therefore, we developed new measures of EOL for use in HD. METHODS: An EOL item pool of 45 items was field tested in 507 individuals with prodromal or manifest HD. Exploratory and confirmatory factor analyses (EFA and CFA, respectively) were conducted to establish unidimensional item pools. Item response theory (IRT) and differential item functioning analyses were applied to the identified unidimensional item pools to select the final items. RESULTS: EFA and CFA supported two separate unidimensional sets of items: Concern with Death and Dying (16 items), and Meaning and Purpose (14 items). IRT and DIF supported the retention of 12 Concern with Death and Dying items and 4 Meaning and Purpose items. IRT data supported the development of both a computer adaptive test (CAT) and a 6-item, static short form for Concern with Death and Dying. CONCLUSION: The HDQLIFE Concern with Death and Dying CAT and corresponding 6-item short form, and the 4-item calibrated HDQLIFE Meaning and Purpose scale demonstrate excellent psychometric properties. These new measures have the potential to provide clinically meaningful information about end-of-life preferences and concerns to clinicians and researchers working with individuals with HD. In addition, these measures may also be relevant and useful for other terminal conditions

Methods for Collecting Milk from Mice

Mouse models offer unique opportunities to study mammary gland biology and lactation. Phenotypes within the mammary glands, especially those caused by genetic modification, often arise during lactation, and their study requires the collection of adequate volumes of milk. We describe two approaches for collecting milk from lactating mice. Both methods are inexpensive, are easy to use in the laboratory or classroom, are non-invasive, and yield adequate volumes of milk for subsequent analyses

Cardiovascular/stroke risk predictive calculators: a comparison between statistical and machine learning models

Background: Statistically derived cardiovascular risk calculators (CVRC) that use conventional risk factors, generally underestimate or overestimate the risk of cardiovascular disease (CVD) or stroke events primarily due to lack of integration of plaque burden. This study investigates the role of machine learning (ML)-based CVD/stroke risk calculators (CVRCML) and compares against statistically derived CVRC (CVRCStat) based on (I) conventional factors or (II) combined conventional with plaque burden (integrated factors). Methods: The proposed study is divided into 3 parts: (I) statistical calculator: initially, the 10-year CVD/stroke risk was computed using 13 types of CVRCStat (without and with plaque burden) and binary risk stratification of the patients was performed using the predefined thresholds and risk classes; (II) ML calculator: using the same risk factors (without and with plaque burden), as adopted in 13 different CVRCStat, the patients were again risk-stratified using CVRCML based on support vector machine (SVM) and finally; (III) both types of calculators were evaluated using AUC based on ROC analysis, which was computed using combination of predicted class and endpoint equivalent to CVD/stroke events. Results: An Institutional Review Board approved 202 patients (156 males and 46 females) of Japanese ethnicity were recruited for this study with a mean age of 69±11 years. The AUC for 13 different types of CVRCStat calculators were: AECRS2.0 (AUC 0.83, P<0.001), QRISK3 (AUC 0.72, P<0.001), WHO (AUC 0.70, P<0.001), ASCVD (AUC 0.67, P<0.001), FRScardio (AUC 0.67, P<0.01), FRSstroke (AUC 0.64, P<0.001), MSRC (AUC 0.63, P=0.03), UKPDS56 (AUC 0.63, P<0.001), NIPPON (AUC 0.63, P<0.001), PROCAM (AUC 0.59, P<0.001), RRS (AUC 0.57, P<0.001), UKPDS60 (AUC 0.53, P<0.001), and SCORE (AUC 0.45, P<0.001), while the AUC for the CVRCML with integrated risk factors (AUC 0.88, P<0.001), a 42% increase in performance. The overall risk-stratification accuracy for the CVRCML with integrated risk factors was 92.52% which was higher compared all the other CVRCStat. Conclusions: ML-based CVD/stroke risk calculator provided a higher predictive ability of 10-year CVD/ stroke compared to the 13 different types of statistically derived risk calculators including integrated model AECRS 2.0

Brain MRI-based Wilson disease tissue classification: An optimised deep transfer learning approach

Wilson's disease (WD) is caused by the excessive accumulation of copper in the brain and liver, leading to death if not diagnosed early. WD shows its prevalence as white matter hyperintensity (WMH) in MRI scans. It is challenging and tedious to classify WD against controls when comparing visually, primarily due to subtle differences in WMH. This Letter presents a computer-aided design-based automated classification strategy that uses optimised transfer learning (TL) utilising two novel paradigms known as (i) MobileNet and (ii) the Visual Geometric Group-19 (VGG-19). Further, the authors benchmark TL systems against a machine learning (ML) paradigm. Using four-fold augmentation, VGG-19 is superior to MobileNet demonstrating accuracy and area under the curve (AUC) pairs as 95.46 ± 7.70%, 0.932 (p < 0.0001) and 86.87 ± 2.23%, 0.871 (p < 0.0001), respectively. Further, MobileNet and VGG-19 showed an improvement of 3.4 and 13.5%, respectively, when benchmarked against the ML-based soft classifier - Random Forest

Is early center-based child care associated with tantrums and unmanageable behavior over time up to school entry?

Background. Existing research suggests that there is a relationship between greater exposure to center-based child care and child behavioral problems though the mechanism for the impact is unclear. However the measure used to document child care has usually been average hours, which may be particularly unreliable in the early months when fewer children are in center care. In addition individual trajectories for behavior difficulties have not been studied. Objective. The purpose of the current study was to examine whether the extent of exposure to center-based child care before two years predicted the trajectory of children’s difficult behavior (i.e., tantrums and unmanageable behavior) from 30 to 51 months controlling for child and maternal characteristics. Method. Data were drawn from UK-based Families, Children and Child Care (FCCC) study (n=1201). Individual growth models were fitted to test the relation between early center-based child care experiences and subsequent difficult behavior. Results. Children with more exposure to center-based care before two had less difficult behavior at 30 months, but more increase over time. Initial levels were predicted by higher difficult temperament and lower verbal ability. Higher difficult temperament and lower family socio-economic status predicted its change over time. Conclusion. Findings suggest that early exposure to center-based care before two years old is a risk factor for subsequent behavior problems especially when children have a longer period of exposure. A possible explanatory process is that child coping strategies to manage frustration are less well developed in a group context, especially when they lag behind in expressive language

Cisplatin-induced emesis: systematic review and meta-analysis of the ferret model and the effects of 5-HT3 receptor antagonists

PURPOSE: The ferret cisplatin emesis model has been used for ~30 years and enabled identification of clinically used anti-emetics. We provide an objective assessment of this model including efficacy of 5-HT(3) receptor antagonists to assess its translational validity. METHODS: A systematic review identified available evidence and was used to perform meta-analyses. RESULTS: Of 182 potentially relevant publications, 115 reported cisplatin-induced emesis in ferrets and 68 were included in the analysis. The majority (n = 53) used a 10 mg kg(−1) dose to induce acute emesis, which peaked after 2 h. More recent studies (n = 11) also used 5 mg kg(−1), which induced a biphasic response peaking at 12 h and 48 h. Overall, 5-HT(3) receptor antagonists reduced cisplatin (5 mg kg(−1)) emesis by 68% (45–91%) during the acute phase (day 1) and by 67% (48–86%) and 53% (38–68%, all P < 0.001), during the delayed phase (days 2, 3). In an analysis focused on the acute phase, the efficacy of ondansetron was dependent on the dosage and observation period but not on the dose of cisplatin. CONCLUSION: Our analysis enabled novel findings to be extracted from the literature including factors which may impact on the applicability of preclinical results to humans. It reveals that the efficacy of ondansetron is similar against low and high doses of cisplatin. Additionally, we showed that 5-HT(3) receptor antagonists have a similar efficacy during acute and delayed emesis, which provides a novel insight into the pharmacology of delayed emesis in the ferret

Abnormal Wnt and PI3Kinase Signaling in the Malformed Intestine of lama5 Deficient Mice

Laminins are major constituents of basement membranes and are essential for tissue homeostasis. Laminin-511 is highly expressed in the intestine and its absence causes severe malformation of the intestine and embryonic lethality. To understand the mechanistic role of laminin-511 in tissue homeostasis, we used RNA profiling of embryonic intestinal tissue of lama5 knockout mice and identified a lama5 specific gene expression signature. By combining cell culture experiments with mediated knockdown approaches, we provide a mechanistic link between laminin α5 gene deficiency and the physiological phenotype. We show that laminin α5 plays a crucial role in both epithelial and mesenchymal cell behavior by inhibiting Wnt and activating PI3K signaling. We conclude that conflicting signals are elicited in the absence of lama5, which alter cell adhesion, migration as well as epithelial and muscle differentiation. Conversely, adhesion to laminin-511 may serve as a potent regulator of known interconnected PI3K/Akt and Wnt signaling pathways. Thus deregulated adhesion to laminin-511 may be instrumental in diseases such as human pathologies of the gut where laminin-511 is abnormally expressed as it is shown here