Who on IRF are you? IRF8 deficiency redirects cDC1 lineage commitment.

Interferon regulatory factor 8 (IRF8) has long been associated with conventional dendritic cell type I (cDC1) development. In a recent study, Lança et al. demonstrate that IRF8 is also crucial in cells already committed to the cDC1 lineage. Here, deletion of IRF8 from the XCR1-expressing pre-cDC1 stage onward leads to a loss of commitment and reprogramming of the cells toward a cDC2-like phenotype.

Psychosocial support and resilience building among health workers in Sierra Leone: interrelations between coping skills, stress levels, and interpersonal relationships.

BACKGROUND: In low- and middle-income countries, a shortage of properly trained, supervised, motivated and equitably distributed health workers often hinder the delivery of lifesaving interventions. Various health workforce bottlenecks can be addressed by tackling well-being and interpersonal relationships of health workers with their colleagues and clients. This paper uses data from the Helping Health Workers Cope (HHWC) project in a rural district of Sierra Leone to achieve three objectives. First, we describe the effect of counseling and psychosocial training on coping skills, stress levels, and provider-provider and provider-client relationships. Second, we examine whether a change in coping skills is associated with a change in relationships. Finally, we qualitatively identify key ways through which the uptake of coping skills is linked to a change in relationships. METHODS: The HHWC project was implemented from February 2012 to June 2013 in Kono district in the Eastern province of Sierra Leone, with the neighboring district of Tonkolili selected as the control site. The evaluation followed a mixed-methods approach, which included a quantitative survey, in-depth interviews and focus group discussions with health workers and clients. Mean values of the variables of interest were compared across sub-populations, and correlation analyses were performed between changes in coping skills, stress levels, and changes in relationships. RESULTS: Overall, the results demonstrate that the HHWC intervention had a positive effect on coping skills, stress levels and provider-provider and provider-client relationships. Furthermore, associations were observed between changes in coping skills and changes in relationships as well as changes in stress management skills and changes in relationships. CONCLUSIONS: Psychosocial education can have major impacts on health worker well-being and the quality of health care delivery. Integrating psychosocial counseling and training interventions into health worker pre-service and in-service curricula would allow the positive effects of the HHWC intervention to be scaled up across Sierra Leone and beyond. A roll out of the HHWC approach alongside health system strengthening initiatives could have major implications for improving health and chances of survival.

Peer-driven quality improvement among health workers and traditional birth attendants in Sierra Leone: linkages between providers' organizational skills and relationships.

BACKGROUND: Sierra Leone has among the poorest maternal and child health indicators in the world and investments in public health have been predominately to increase demand for services, with fewer initiatives targeting supply side factors that influence health workers' work environment. This paper uses data from the Quality Circles project in a rural district of Sierra Leone to achieve three objectives. First, we examine the effect of the intervention on organizational skills and relationships among coworkers as well as between health workers and traditional birth attendants. Second, we examine whether changes in organizational skills are associated with changes in relationships among and between formal and informal health providers and between health providers and clients. Third, we aim to further understand these changes through the perspectives of health workers and traditional birth attendants. METHODS: The Quality Circles project was implemented in Kailahun District in the Eastern province of Sierra Leone from August 2011 to June 2013, with adjacent Tonkolili District serving as the control site. Using a mixed-methods approach, the evaluation included a quantitative survey, in-depth interviews and focus group discussions with health workers and traditional birth attendants. Mean values of the variables of interest were compared across sub-populations, and correlation analyses were performed between changes in organizational skills and changes in relationships. RESULTS: The results demonstrate that the Quality Circles intervention had positive effects on organizational skills and relationships. Furthermore, improvements in all organizational skill variables - problem-solving, strategizing and negotiation skills - were strongly associated with a change in the overall relationship variable. CONCLUSIONS: The Quality Circles approach has the potential to support health workers to improve their organizational skills and relationships, which in turn can contribute to improving the interpersonal dimensions of the quality of care in low-resource contexts. This method brings together peers in a structured process for constructive group work and individual skill development, which are important in low-resource contexts where active participation and resourcefulness of health workers can also contribute to better health service delivery.

Single-cell phenotypes of peripheral blood immune cells in early and late stages of non-alcoholic fatty liver disease.

BACKGROUND/AIMS: Immune and inflammatory cells respond to multiple pathological hits in the development of nonalcoholic steatohepatitis (NASH) and fibrosis. Relatively little is known about how their type and function change through the non-alcoholic fatty liver disease (NAFLD) spectrum. Here we used multi-dimensional mass cytometry and a tailored bioinformatic approach to study circulating immune cells sampled from healthy individuals and people with NAFLD. METHODS: Cytometry by time of flight using 36 metal-conjugated antibodies was applied to peripheral blood mononuclear cells (PBMCs) from biopsy-proven NASH fibrosis (late disease), steatosis (early disease), and healthy patients. Supervised and unsupervised analyses were used, findings confirmed, and mechanisms assessed using independent healthy and disease PBMC samples. RESULTS: Of 36 PBMC clusters, 21 changed between controls and disease samples. Significant differences were observed between diseases stages with changes in T cells and myeloid cells throughout disease and B cell changes in late stages. Semi-supervised gating and re-clustering showed that disease stages were associated with fewer monocytes with active signalling and more inactive NK cells; B and T cells bearing activation markers were reduced in late stages, while B cells bearing co-stimulatory molecules were increased. Functionally, disease states were associated with fewer activated mucosal-associated invariant T cells and reduced toll-like receptor-mediated cytokine production in late disease. CONCLUSION: A range of innate and adaptive immune changes begin early in NAFLD, and disease stages are associated with a functionally less active phenotype compared to controls. Further study of the immune response in NAFLD spectrum may give insight into mechanisms of disease with potential clinical application.

G protein-coupled receptor kinase 5 regulates thrombin signaling in platelets via PAR-1.

The interindividual variation in the functional response of platelets to activation by agonists is heritable. Genome-wide association studies (GWASs) of quantitative measures of platelet function have identified fewer than 20 distinctly associated variants, some with unknown mechanisms. Here, we report GWASs of pathway-specific functional responses to agonism by adenosine 5'-diphosphate, a glycoprotein VI-specific collagen mimetic, and thrombin receptor-agonist peptides, each specific to 1 of the G protein-coupled receptors PAR-1 and PAR-4, in subsets of 1562 individuals. We identified an association (P = 2.75 × 10-40) between a common intronic variant, rs10886430, in the G protein-coupled receptor kinase 5 gene (GRK5) and the sensitivity of platelets to activate through PAR-1. The variant resides in a megakaryocyte-specific enhancer that is bound by the transcription factors GATA1 and MEIS1. The minor allele (G) is associated with fewer GRK5 transcripts in platelets and the greater sensitivity of platelets to activate through PAR-1. We show that thrombin-mediated activation of human platelets causes binding of GRK5 to PAR-1 and that deletion of the mouse homolog Grk5 enhances thrombin-induced platelet activation sensitivity and increases platelet accumulation at the site of vascular injury. This corroborates evidence that the human G allele of rs10886430 is associated with a greater risk for cardiovascular disease. In summary, by combining the results of pathway-specific GWASs and expression quantitative trait locus studies in humans with the results from platelet function studies in Grk5-/- mice, we obtain evidence that GRK5 regulates the human platelet response to thrombin via the PAR-1 pathway.

Effects of central and peripheral administration of an acute-phase protein, α-1-acid-glycoprotein, on feed intake and rectal temperature in sheep.

In rodents, an acute-phase protein, α-1-acid-glycoprotein (AGP), was shown to provide a link between inflammation and suppression of feed intake by acting as a leptin receptor agonist. The objective of this study was to determine the effects of AGP on feed intake and rectal temperature in sheep. Ewes were ovariectomized, implanted with a cannula into a lateral ventricle of the brain, and kept indoors in individual pens. Feed intake and rectal temperature were determined for sheep in all experiments. In the first experiment, ewes (n = 4) received 1 of 4 treatments [0 (control), 0.012 (low), 0.06 (medium), or 0.30 (high) mg/kg BW AGP] into the lateral ventricle (ICV). All sheep received all treatments in a Latin square design balanced for carryover effects with 10 d between treatments. In the second experiment, ewes (n = 10) received 1 of 2 treatments (0 and 3 mg/kg BW of AGP) intravenously (IV) in a completely randomized design. In the third experiment, ewes (n = 19) received peripheral treatments (IV) of an antipyretic [0 (control) or 2.2 mg/kg BW flunixin meglumine (FLU)] 30 min before receiving central AGP [0 (control) or 0.3 mg/kg BW of AGP] in a completely randomized design. All data were analyzed using a mixed model analysis of variance and tested for effects of treatment, time, and the interaction of treatment and time. Cumulative 48-h feed intake after administration of treatments was also determined. In the first experiment, there was no effect of ICV treatment (P = 0.37) on feed intake rate or on cumulative feed intake (P = 0.31). There was an effect of ICV treatment (P = 0.002) on rectal temperatures, which were greater (P < 0.05) after the high dose of centrally administered AGP. In the second experiment, there was no effect of AGP administration IV on feed intake rate (P = 0.98), on cumulative feed intake (P = 0.41) or on rectal temperature (P = 0.71). In the third experiment, there was an effect of central AGP treatment (P < 0.0001) and an interaction of central AGP and time (P < 0.0001) on rectal temperature, whereas FLU had no effect (P = 0.93), demonstrating that AGP increased rectal temperatures regardless of antipyretic treatment. These results indicate that central AGP increases rectal temperature in sheep by pathways that do not involve prostaglandins. Further research is needed to determine whether AGP may be an important integrator of energy balance and inflammation.

ADAM17-Mediated Reduction in CD14++CD16+ Monocytes ex vivo and Reduction in Intermediate Monocytes With Immune Paresis in Acute Pancreatitis and Acute Alcoholic Hepatitis.

Impaired immune responses and increased susceptibility to infection characterize acute inflammatory conditions such as pancreatitis and alcoholic hepatitis and are major causes of morbidity and mortality. However, the mechanisms that drive this apparent immune paresis remain poorly understood. Monocytes mediate host responses to damage and pathogens in health and disease, and three subsets of monocytes have been defined based on CD14 and CD16 expression. We sought to determine the changes in monocyte subsets in acute pancreatitis (AP) and acute alcoholic hepatitis (AAH), together with functional consequences and mechanisms that underlie this change. Peripheral blood mononuclear cells (PBMCs) from patients with AP or AAH were compared with healthy controls. Monocyte subsets were defined by HLA-DR, CD14, and CD16 expression. Changes in surface and intracellular protein expression and phosphorylation were determined by flow cytometry. Phenotype and function were assessed following stimulation with lipopolysaccharide (LPS) or other agonists in the presence of specific inhibitors of TNFα and a disintegrin and metalloproteinase 17 (ADAM17). Patients with AP and AAH had reduced CD14++CD16+ intermediate monocytes compared to controls. Reduction of intermediate monocytes was recapitulated ex vivo by stimulating healthy control PBMCs with Toll-like receptor (TLR) agonists LPS, flagellin or polyinosilic:polycytidylic acid (poly I:C). Stimulation caused shedding of CD14 and CD16, which could be reversed using the ADAM17 inhibitor, TMI005 but not direct inhibitors of TNFα, a known ADAM17-target. Culturing PBMCs from healthy controls resulted in expansion of intermediate monocytes, which did not occur when LPS was in the culture medium. Cultured intermediate monocytes showed reduced expression of CX3CR1, CCR2, TLR4, and TLR5. We found reduced migratory responses, intracellular signaling and pro-inflammatory cytokine production, and increased expression of IL-10. Stimulation with TLR agonists results in ADAM17-mediated shedding of phenotypic markers from CD16+ monocytes, leading to apparent "loss" of intermediate monocytes. Reduction in CD14++CD16- monocytes and increased CD14++CD16+ is associated with altered responses in functional assays ex vivo. Patients with AP and AAH had reduced proportions of CD14++CD16+ monocytes and reduced phosphorylation of NFκB and IL-6 production in response to bacterial LPS. Together, these processes may contribute to the susceptibility to infection observed in AP and AAH.