A genetically inducible endothelial niche enables vascularization of human kidney organoids with multilineage maturation and emergence of renin expressing cells.

Vascularization plays a critical role in organ maturation and cell type development. Drug discovery, organ mimicry, and ultimately transplantation hinge on achieving robust vascularization of in vitro engineered organs. Here, focusing on human kidney organoids, we overcame this hurdle by combining a human induced pluripotent stem cell (iPSC) line containing an inducible ETS translocation variant 2 (ETV2) (a transcription factor playing a role in endothelial cell development) that directs endothelial differentiation in vitro, with a non-transgenic iPSC line in suspension organoid culture. The resulting human kidney organoids show extensive endothelialization with a cellular identity most closely related to human kidney endothelia. Endothelialized kidney organoids also show increased maturation of nephron structures, an associated fenestrated endothelium with de novo formation of glomerular and venous subtypes, and the emergence of drug-responsive renin expressing cells. The creation of an engineered vascular niche capable of improving kidney organoid maturation and cell type complexity is a significant step forward in the path to clinical translation. Thus, incorporation of an engineered endothelial niche into a previously published kidney organoid protocol allowed the orthogonal differentiation of endothelial and parenchymal cell types, demonstrating the potential for applicability to other basic and translational organoid studies.

'Love with Less Salt': evaluation of a sodium reduction mass media campaign in China.

OBJECTIVE: This study examines the impact of a salt reduction campaign on knowledge, attitudes, intentions, behaviours and barriers to behaviour change relating to salt consumption in two provinces of China. METHODS: In 2019, the 'Love with Less Salt' campaign ran on China Central Television and on local television channels in Shandong and Anhui provinces. Data for this study come from two representative household surveys conducted among a sample of adults aged 25-65 years in Shandong and Anhui provinces: precampaign (n=2000) and postcampaign (n=2015). Logistic regression was performed to estimate the effects of the campaign on knowledge, attitudes, intentions, behaviours and barriers to behaviour change. RESULTS: Overall, 13% of postcampaign respondents recalled seeing the campaign, and reactions towards the campaign were positive. Postcampaign respondents were more likely to plan to reduce their purchase of foods high in salt than precampaign respondents (OR=1.45, p=<0.05). Campaign-aware respondents were significantly more likely than campaign-unaware respondents to report higher levels of knowledge, attitudes and behaviours regarding salt reduction. CONCLUSIONS: Findings reveal that salt reduction mass media campaigns can be an effective public health tool to support efforts to reduce salt consumption in China. Continued and sustained mass media investments are likely to be effective in addressing high salt consumption nationwide.

What makes an effective antismoking campaign - insights from the trenches.

OBJECTIVES: This paper describes traditionally effective approaches for anti-smoking mass media advertising and explores challenges and future directions for campaign planning. The changing characteristics of the current smoking population and media landscape are examined. Type of program or service: Anti-smoking mass media advertising campaigns. METHODS: We present a commentary on the established creative and media strategies proven to be effective in prompting quit attempts among smokers, discuss new challenges facing anti-smoking campaign managers today and propose considerations for the future. RESULTS: Although evidence of effective approaches for tobacco control messaging and execution remains clear, the media landscape in Australia has changed dramatically in recent years with some audiences moving away from frequent and heavy television consumption towards online platforms and digital media channels. In addition, as smoking rates continue to fall, characteristics of current smokers are becoming increasingly relevant considerations for anti-smoking messaging and placement within a media environment that is becoming more expensive and fragmented. Funding anti-smoking advertising at the levels required to effectively prompt and maintain smoking cessation remains a high priority considering the extensive social and economic costs of smoking. LESSONS LEARNT: Although it is known that hard-hitting, emotional and/or testimonial anti-smoking advertisements can be effective in prompting quit attempts, optimal media channel selection and media mix for reaching and engaging audience segments is dynamic in an ever-changing media landscape. Targeting media channels popular with lower socio-economic status (SES) smokers can efficiently achieve wide population exposure as well as effectively reaching population groups with higher smoking prevalence. A collaborative approach between health professionals, researchers, creative directors and media buyers is required to ensure advertising materials are communicating the right message for the audience, and that it is being delivered effectively.

Is this health campaign really social marketing? A checklist to help you decide.

ISSUE ADDRESSED: Social marketing (SM) campaigns can be a powerful disease prevention and health promotion strategy but health-related campaigns may simply focus on the "promotions" communication activities and exclude other key characteristics of the SM approach. This paper describes the application of a checklist for identifying which lifestyle-related chronic disease prevention campaigns reported as SM actually represent key SM principles and practice. METHODS: A checklist of SM criteria was developed, reviewed and refined by SM and mass media campaign experts. Papers identified in searches for "social marketing" and "mass media" for obesity, diet and physical activity campaigns in the health literature were classified using the checklist. RESULTS: Using the checklist, 66.6% of papers identified in the "SM" search and 39% of papers identified from the "mass media" search were classified as SM campaigns. Inter-rater agreement for classification using the abstract only was 92.1%. CONCLUSIONS: Health-related campaigns that self-identify as "social marketing" or "mass media" may not include the key characteristics of a SM approach. Published literature can provide useful guidance for developing and evaluating health-related SM campaigns, but health promotion professionals need to be able to identify what actually comprises SM in practice. SO WHAT?: SM could be a valuable strategy in comprehensive health promotion interventions, but it is often difficult for non-experts to identify published campaigns that represent a true SM approach. This paper describes the application of a checklist to assist policy makers and practitioners in appraising evidence from campaigns reflecting actual SM in practice. The checklist could also guide reporting on SM campaigns.

A Systematic Search and Review of Adult-Targeted Overweight and Obesity Prevention Mass Media Campaigns and Their Evaluation: 2000-2017.

Mass media campaigns are a commonly used strategy in public health. However, no review has assessed whether the design and evaluation of overweight and obesity campaigns meets best practice recommendations. This study aimed to fill this gap. We systematically searched five databases for peer-reviewed articles describing adult-targeted obesity mass media campaigns published between 2000 and 2017, complemented by reference list searches and contact with authors and agencies responsible for the campaigns. We extracted data on campaign design, implementation, and evaluation from eligible publications and conducted a qualitative review of 29 publications reporting on 14 campaigns. We found a need for formative research with target audiences to ensure campaigns focus on the most salient issues. Further, we noted that most campaigns targeted individual behaviors, despite calls for campaigns to also focus upstream and to address social determinants of obesity. Television was the dominant communication channel but, with the rapid advance of digital media, evaluation of other channels, such as social media, is increasingly important. Finally, although evaluation methods varied in quality, the evidence suggests that campaigns can have an impact on intermediate outcomes, such as knowledge and attitudes. However, evidence is still limited as to whether campaigns can influence behavior change.

An H3K9/S10 methyl-phospho switch modulates Polycomb and Pol II binding at repressed genes during differentiation.

Methylated histones H3K9 and H3K27 are canonical epigenetic silencing modifications in metazoan organisms, but the relationship between the two modifications has not been well characterized. H3K9me3 coexists with H3K27me3 in pluripotent and differentiated cells. However, we find that the functioning of H3K9me3 is altered by H3S10 phosphorylation in differentiated postmitotic osteoblasts and cycling B cells. Deposition of H3K9me3/S10ph at silent genes is partially mediated by the mitogen- and stress-activated kinases (MSK1/2) and the Aurora B kinase. Acquisition of H3K9me3/S10ph during differentiation correlates with loss of paused S5 phosphorylated RNA polymerase II, which is present on Polycomb-regulated genes in embryonic stem cells. Reduction of the levels of H3K9me3/S10ph by kinase inhibition results in increased binding of RNAPIIS5ph and the H3K27 methyltransferase Ezh1 at silent promoters. Our results provide evidence of a novel developmentally regulated methyl-phospho switch that modulates Polycomb regulation in differentiated cells and stabilizes repressed states.

Naive pluripotency is associated with global DNA hypomethylation.

Naive pluripotent embryonic stem cells (ESCs) and embryonic germ cells (EGCs) are derived from the preimplantation epiblast and primordial germ cells (PGCs), respectively. We investigated whether differences exist between ESCs and EGCs, in view of their distinct developmental origins. PGCs are programmed to undergo global DNA demethylation; however, we find that EGCs and ESCs exhibit equivalent global DNA methylation levels. Inhibition of MEK and Gsk3b by 2i conditions leads to pronounced reduction in DNA methylation in both cell types. This is driven by Prdm14 and is associated with downregulation of Dnmt3a and Dnmt3b. However, genomic imprints are maintained in 2i, and we report derivation of EGCs with intact genomic imprints. Collectively, our findings establish that culture in 2i instills a naive pluripotent state with a distinctive epigenetic configuration that parallels molecular features observed in both the preimplantation epiblast and nascent PGCs.

Polycomb associates genome-wide with a specific RNA polymerase II variant, and regulates metabolic genes in ESCs.

Polycomb repressor complexes (PRCs) are important chromatin modifiers fundamentally implicated in pluripotency and cancer. Polycomb silencing in embryonic stem cells (ESCs) can be accompanied by active chromatin and primed RNA polymerase II (RNAPII), but the relationship between PRCs and RNAPII remains unclear genome-wide. We mapped PRC repression markers and four RNAPII states in ESCs using ChIP-seq, and found that PRC targets exhibit a range of RNAPII variants. First, developmental PRC targets are bound by unproductive RNAPII (S5p(+)S7p(-)S2p(-)) genome-wide. Sequential ChIP, Ring1B depletion, and genome-wide correlations show that PRCs and RNAPII-S5p physically bind to the same chromatin and functionally synergize. Second, we identify a cohort of genes marked by PRC and elongating RNAPII (S5p(+)S7p(+)S2p(+)); they produce mRNA and protein, and their expression increases upon PRC1 knockdown. We show that this group of PRC targets switches between active and PRC-repressed states within the ESC population, and that many have roles in metabolism.

A role for cohesin in T-cell-receptor rearrangement and thymocyte differentiation.

Cohesin enables post-replicative DNA repair and chromosome segregation by holding sister chromatids together from the time of DNA replication in S phase until mitosis. There is growing evidence that cohesin also forms long-range chromosomal cis-interactions and may regulate gene expression in association with CTCF, mediator or tissue-specific transcription factors. Human cohesinopathies such as Cornelia de Lange syndrome are thought to result from impaired non-canonical cohesin functions, but a clear distinction between the cell-division-related and cell-division-independent functions of cohesion--as exemplified in Drosophila--has not been demonstrated in vertebrate systems. To address this, here we deleted the cohesin locus Rad21 in mouse thymocytes at a time in development when these cells stop cycling and rearrange their T-cell receptor (TCR) α locus (Tcra). Rad21-deficient thymocytes had a normal lifespan and retained the ability to differentiate, albeit with reduced efficiency. Loss of Rad21 led to defective chromatin architecture at the Tcra locus, where cohesion-binding sites flank the TEA promoter and the Eα enhancer, and demarcate Tcra from interspersed Tcrd elements and neighbouring housekeeping genes. Cohesin was required for long-range promoter-enhancer interactions, Tcra transcription, H3K4me3 histone modifications that recruit the recombination machinery and Tcra rearrangement. Provision of pre-rearranged TCR transgenes largely rescued thymocyte differentiation, demonstrating that among thousands of potential target genes across the genome, defective Tcra rearrangement was limiting for the differentiation of cohesin-deficient thymocytes. These findings firmly establish a cell-division-independent role for cohesin in Tcra locus rearrangement and provide a comprehensive account of the mechanisms by which cohesin enables cellular differentiation in a well-characterized mammalian system.

Culturally and linguistically diverse population health social marketing campaigns in Australia: a consideration of evidence and related evaluation issues.

ISSUE ADDRESSED: This paper describes a review of population health social marketing campaigns targeting culturally and linguistically diverse (CLD) communities in Australia in order to identify characteristics of effective CLD campaigns. METHODS: Literature on CLD population health social marketing was identified from electronic searches of databases in August 2004. At the same time, the grey literature was examined by searching the Internet and talking to Australian experts in the fields of CLD social marketing and CLD research. RESULTS: Eight studies met the search criteria, four from the published literature. Two studies that employed prepost evaluation designs provided tentative support for the potential efficacy of CLD social marketing strategies. The remaining studies did not allow for causal attribution as they used post-campaign only or process evaluations. Studies did, however, show that CLD communities access campaign-related information from both mainstream and ethnic media channels. In addition, Vietnamese respondents were more likely to access campaign messages through ethnic radio and Chinese respondents through ethnic press. CONCLUSIONS: There is insufficient evidence to clearly identify the characteristics of effective CLD campaigns. Campaign evaluation designs used to evaluate social marketing strategies targeting CLD communities in Australia are generally weak, but there is tentative evidence supporting the potential efficacy of these strategies in some Australian settings.

Collecting duct-specific gene inactivation of alphaENaC in the mouse kidney does not impair sodium and potassium balance.

Aldosterone controls the final sodium reabsorption and potassium secretion in the kidney by regulating the activity of the epithelial sodium channel (ENaC) in the aldosterone-sensitive distal nephron (ASDN). ASDN consists of the last portion of the distal convoluted tubule (late DCT), the connecting tubule (CNT), and the collecting duct (CD) (i.e., the cortical CD [CCD] and the medullary CD [MCD]). It has been proposed that the control of sodium transport in the CCD is essential for achieving sodium and potassium balance. We have tested this hypothesis by inactivating the alpha subunit of ENaC in the CD but leaving ENaC expression in the late DCT and CNT intact. Under salt restriction or under aldosterone infusion, whole-cell voltage clamp of principal cells of CCD showed no detectable ENaC activity, whereas large amiloride-sensitive currents were observed in control littermates. The animals survive well and are able to maintain sodium and potassium balance, even when challenged by salt restriction, water deprivation, or potassium loading. We conclude that the expression of ENaC in the CD is not a prerequisite for achieving sodium and potassium balance in mice. This stresses the importance of more proximal nephron segments (late DCT/CNT) to achieve sodium and potassium balance.