A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis.

Phagocytic clearance of dying cells, termed efferocytosis, is essential for maintaining tissue homeostasis, yet our understanding of efferocytosis regulation remains incomplete. Here we perform a FACS-based, genome-wide CRISPR knockout screen in primary mouse macrophages to search for novel regulators of efferocytosis. The results show that Wdfy3 knockout in macrophages specifically impairs uptake, but not binding, of apoptotic cells due to defective actin disassembly. Additionally, WDFY3 interacts with GABARAP, thus facilitating LC3 lipidation and subsequent lysosomal acidification to permit the degradation of apoptotic cell components. Mechanistically, while the C-terminus of WDFY3 is sufficient to rescue the impaired degradation induced by Wdfy3 knockout, full-length WDFY3 is required to reconstitute the uptake of apoptotic cells. Finally, WDFY3 is also required for efficient efferocytosis in vivo in mice and in vitro in primary human macrophages. This work thus expands our knowledge of the mechanisms of macrophage efferocytosis, as well as supports genome-wide CRISPR screen as a platform for interrogating complex functional phenotypes in primary macrophages.

A Collaborative Team Teaching Model for a MSW Capstone Course.

This exploratory study was embedded in a formative process for the purposes of improving content delivery to an evidence-based practice class, and improving students' performance on a comprehensive exam. A learning and teaching model was utilized by faculty from a three-university collaborative graduate social work program to examine the extent to which course texts and assignments explicitly supported the process, application, and evaluation of evidence-based practices. The model was grounded in a collaborative culture, allowing each faculty to share their collective skills and knowledge across a range of practice settings as they revised the course curriculum. As a result, faculty found they had created a unique community that allowed a wider context for learning and professional development that translated into the classroom. Students enrolled in the revised course across all three universities showed improvement on the comprehensive exam. When faculty themselves invest in collaborative learning and teaching, students benefit.

Lack of effects of guanfacine on executive and memory functions in healthy male volunteers.

RATIONALE: Guanfacine is an alpha2-adrenergic receptor agonist that has been shown to have beneficial effects on working memory and attentional functions in monkeys and in patients with attention deficit hyperactivity disorder. OBJECTIVES: The aim of this study was to further investigate the cognitive-enhancing properties of guanfacine using an established battery of tasks measuring executive and memory functions. METHODS: Sixty healthy male volunteers were randomised into three groups. Cognitive testing was performed from +2 to +4 h after double-blind administration of a single oral dose of 1 or 2 mg of guanfacine or placebo. RESULTS: Systolic blood pressure was significantly reduced by both doses of guanfacine at the end of the testing session. There were no statistically significant effects on any of the cognitive measures. Two trend effects were observed with poorer performance on digit span backward and slower 'Go' reaction times after guanfacine. CONCLUSION: This study found no improvement of prefrontal memory or executive functions after guanfacine. Negative effects on blood pressure and trend effects on digit span backward and go reaction time indicate a mild sedative effect of guanfacine at these doses, possibly via mechanisms of autoreceptor down-regulation.

In vitro uptake and metabolism of testosterone by goldfish, Carassius auratus, mesenteric adipose tissue.

Mesenteric adipose tissue was removed from adult goldfish, Carassius auratus, and incubated in vitro with (1alpha,2alpha)-3[H]testosterone (T). Total radioactivity in the medium decreased and tissue total radioactivity increased in a time-dependent fashion between 1 and 6 h of incubation with maximum uptake occurring between 4 and 6 h. After ether extraction and thin layer chromatography the amount of radioactivity comigrating with authentic T standard decreased over time in both medium and tissue samples. Radioactivity in the aqueous fraction remaining after ether extraction increased in a time-dependent manner, suggesting the presence of water-soluble conjugated steroids. Acid hydrolysis of the aqueous fraction yielded radioactivity that principally comigrated with T standard following a second ether extraction and TLC, along with a small, unidentified peak of poorly migrating radioactivity. Aromatization of T was assessed after charcoal stripping samples and measuring radioactivity remaining in the form of 3H(2)O. Small but significant amounts of 3H(2)O were present in both incubation medium and tissue samples. Incubation with the aromatase inhibitor 4-androsten-4-ol-3,17-dione acetate (ATD) decreased the formation of 3H(2)O in a dose-dependent manner. These results suggest that goldfish mesenteric adipose tissue is capable of converting T to several metabolites including water-soluble conjugates and estrogen.