HIF prolyl hydroxylase 2/3 deletion disrupts astrocytic integrity and exacerbates neuroinflammation

Astrocytes constitute the parenchymal border of the blood-brain barrier (BBB), modulate the exchange of soluble and cellular elements, and are essential for neuronal metabolic support. Thus, astrocytes critically influence neuronal network integrity. In hypoxia, astrocytes upregulate a transcriptional program that has been shown to boost neuroprotection in several models of neurological diseases. We investigated transgenic mice with astrocyte-specific activation of the hypoxia-response program by deleting the oxygen sensors, HIF prolyl-hydroxylase domains 2 and 3 (Phd2/3). We induced astrocytic Phd2/3 deletion after onset of clinical signs in experimental autoimmune encephalomyelitis (EAE) that led to an exacerbation of the disease mediated by massive immune cell infiltration. We found that Phd2/3-ko astrocytes, though expressing a neuroprotective signature, exhibited a gradual loss of gap-junctional Connexin-43 (Cx43), which was induced by vascular endothelial growth factor-alpha (Vegf-a) expression. These results provide mechanistic insights into astrocyte biology, their critical role in hypoxic states, and in chronic inflammatory CNS diseases

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2,3,4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Comparison of RNA isolation procedures for analysis of adult murine brain and spinal cord astrocytes

BACKGROUND: Molecular analyses of cell populations and single cells have been instrumental in the advancement of our understanding of the physiology and pathologic processes of the nervous system. However, the limitation of these methods is the dependence on a gentle, efficient and specific enrichment procedure for the target cell population. In particular, this has been challenging for tightly interconnected cells, for example central nervous system (CNS) endogenous cells such as astrocytes. NEW METHOD: Here we adopted one of the most common methods of cell extraction, namely, enzymatic tissue digestion followed by fluorescence-activated cell sorting (FACS) of individual cells. We evaluated different enzymatic/mechanical tissue dissociation procedures and analyzed different astrocyte lineage transgenic models. Furthermore, we compared the cell extraction efficiency from spinal cord vs. brain. RESULTS: Enzymatic digestion of CNS tissue of Glast-Cre(ERT2)xtdTomato(fl/fl) or Aldh-Cre(ERT2)xtdTomato(fl/fl) followed by FACS resulted in highly purified astrocytes. Automated tissue digestion strongly improved the isolated cell numbers. Aldh1l1-Cre(ERT2) identified more astrocytes than Glast-Cre(ERT2); isolation from brain yields higher numbers than from spinal cord. COMPARISON WITH EXISTING METHODS: We compared the efficiency and purity of the enzymatic dissociation/FACS approach with a more modern procedure consisting of tissue homogenization followed by translating ribosome affinity purification (TRAP). CONCLUSION: We found that both methods result in highly enriched astrocytic RNA. However, only TRAP isolation resulted in reliably detectable RNA concentrations from spinal cord tissue on a single animal level. Depending on the aim of the study both methods have advantages and disadvantages but both are acceptable for astrocytic RNA analysis

Beyond Spanish: Competencies for SLPs Working with Children from Diverse Cultures

This session is developed by, and presenters invited by Issues in Culturally and Linguistically Diverse Populations. This session will support SLPs’competencies for working with children with speech-language disorders across different cultural contexts. General principles and resources will be supplemented with examples from New York, Ghana, Bolivia, Brazil, Navajo Nation, China (Hong Kong), Jamaica, and Canada (Québec)

Beyond Spanish: Competencies for SLPs Working with Children from Diverse Cultures

This session is developed by, and presenters invited by Issues in Culturally and Linguistically Diverse Populations. This session will support SLPs’competencies for working with children with speech-language disorders across different cultural contexts. General principles and resources will be supplemented with examples from New York, Ghana, Bolivia, Brazil, Navajo Nation, China (Hong Kong), Jamaica, and Canada (Québec)

MULTIWAVELENGTH MONITORING OF THE NARROW-LINE SEYFERT 1 GALAXY ARAKELIAN 564. III. OPTICAL OBSERVATIONS AND THE OPTICAL--UV--X-RAY CONNECTION

We present the results of a 2 yr long optical monitoring program of the narrow-line Seyfert 1 galaxy Ark 564. The majority of this monitoring project was also covered by X-ray observations (RXT E), and for a period of ~50 days, we observed the galaxy in UV (HST) and X-rays (RXTE and ASCA) simultaneously with the ground-based observations. Rapid and large-amplitude variations seen in the X-ray band, on a daily and hourly timescale, were not detected at optical and UV wavelengths, which in turn exhibited much lower variability either on short (1 day) or long (several months) timescales. The only significant optical variations can be described as two 2È4 day events with ~10% flux variations. We detect no significant optical line variations and thus cannot infer a reverberation size for the broad-line region. Similarly, the large X-ray variations seem to vanish when the light curve is smoothed over a period of 30 days. The UV continuum follows the X-rays with a lag of ~0.4 days, and the optical band lags the UV band by ~2 days. No signiÐcant correlation was found between the entire X-ray data set and the optical band. Focusing on a 20 day interval around the strongest optical event we detect a significant X-ray--optical correlation with similar events seen in the UV and X-rays. Our data are consistent with reprocessing models on the grounds of the energy emitted in this single event. However, several large X-ray flares produced no corresponding optical emission