Rules of Engagement: Journalists’ attitudes to industry influence in health news reporting.

Health-related industries use a variety of methods to influence health news, including the formation and maintenance of direct relationships with journalists. These interactions have the potential to subvert news reporting such that it comes to serve the interests of industry in promoting their products, rather than the public interest in critical and accurate news and information. Here we report the findings of qualitative interviews conducted in Sydney, Australia, in which we examined journalists’ experiences of, and attitudes towards, their relationships with health-related industries. Participants’ belief in their ability to manage industry influence and their perceptions of what it means to be unduly influenced by industry raise important concerns relating to the psychology of influence and the realities of power relationships between industry and journalists. The analysis also indicates ways in which concerned academics and working journalists might establish more fruitful dialogue regarding the role of industry in health-related news and the extent to which increased regulation of journalist-industry relationships might be needed.NHMR

Loss of Krüppel-like factor 9 deregulates both physiological gene expression and development

Abstract Krüppel-like factor 9 (Klf9) is a ubiquitously expressed transcription factor that is a feedforward regulator of multiple stress-responsive and endocrine signaling pathways. We previously described how loss of Klf9 function affects the transcriptome of zebrafish larvae sampled at a single time point 5 days post-fertilization (dpf). However, klf9 expression oscillates diurnally, and the sampled time point corresponded to its expression nadir. To determine if the transcriptomic effects of the klf9 −/− mutation vary with time of day, we performed bulk RNA-seq on 5 dpf zebrafish embryos sampled at three timepoints encompassing the predawn peak and midmorning nadir of klf9 expression. We found that while the major effects of the klf9 −/− mutation that we reported previously are robust to time of day, the mutation has additional effects that manifest only at the predawn time point. We used a published single-cell atlas of zebrafish development to associate the effects of the klf9 −/− mutation with different cell types and found that the mutation increased mRNA associated with digestive organs (liver, pancreas, and intestine) and decreased mRNA associated with differentiating neurons and blood. Measurements from confocally-imaged larvae suggest that overrepresentation of liver mRNA in klf9 −/− mutants is due to development of enlarged livers

Screen for regulators of P-granule accumulation.

<p>A) Screening strategy to isolate mutants with enlarged PGL-1::GFP granules. PGL-1::GFP worms were mutagenized, 2000 F1 progeny were cloned to individual plates, and F2 progeny were screened for homozygous (m/m) mutants with enlarged P granules B) PGL-1::GFP expression in the gonad arm of parental (P0) and mutant strains. C) Relative PGL-1 intensity in mutant germlines normalized to the parental control. D) Increased PGL-1::GFP size in mutants correlates with increased PGL-1 intensity.</p

Enlarged P-granule alleles from screen.

<p>Enlarged P-granule alleles from screen.</p

Gene expression analysis of <i>elli-1</i>.

<p>A) Volcano plot showing the fold change and significance of gene expression in <i>elli-1</i>. Significantly regulated genes above or below 1.2-fold are shown in red, with a subset of soma-enriched genes [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006611#pgen.1006611.ref043" target="_blank">43</a>] in green (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006611#pgen.1006611.s001" target="_blank">S1 Table</a>). B) Proportional Venn diagrams comparing overlap between the 1079 <i>elli-1</i> regulated genes and previously published <i>csr-1</i> and <i>pgl-1</i> regulated [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006611#pgen.1006611.ref013" target="_blank">13</a>], CSR-1 target [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006611#pgen.1006611.ref004" target="_blank">4</a>], and soma and germline enriched [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006611#pgen.1006611.ref043" target="_blank">43</a>] datasets. C) Increased average expression of CSR-1 complex and core P-granule components in the <i>elli-1</i> expression array (black) and <i>elli-1</i> qRTPCR (green). Pink line indicates the arbitrary 1.2-fold increased expression cutoff. Increased <i>ego-1</i> and <i>ekl-1</i> expression was statistically significant, but under the 1.2-fold cutoff. D) <i>elli-1</i> Volcano plot showing increased expression of genes required for RNAi-dependent gene silencing (green).</p

<i>elli-1</i> expression.

<p>A) Fluorescence In Situ Hybridization (FISH) of <i>elli-1</i> mRNA (red) shows germline expression. DAPI/DNA is blue. B) <i>elli-1</i>::<i>GFP</i>::<i>3xFLAG</i> showing diffuse cytoplasmic expression with some ELLI-1 foci in a dissected germline. 4.3% of ELLI-1 foci are docked to P granules (arrowheads), but most of these foci are in the central shared cytoplasm of the rachis. C) ELLI-1::GFP partially overlaps with the expression of the P-body component CGH-1 (red), primarily in the rachis instead of at the nuclear periphery of germ cells. D) <i>elli-1</i>::<i>GFP</i>::<i>3xFLAG</i> expression in primordial germ cells (arrows) during embryogenesis and the first larval stage. E) Embryonic lethality associated in <i>elli-1</i> worms.</p

Synthetic sterility and RNAi deficiency in <i>elli-1</i>.

<p>A) Larval arrest on control (+) and <i>his-44</i> (-) RNAi showing <i>elli-1(sam3)</i> enhances the RNAi resistant phenotype of <i>drh-3(sam27)</i> at permissive temperature (20°C). Same colored box plots represent duplicate experiments performed on different weeks. B) Brood sizes show that <i>elli-1(sam3)</i> enhances sterility (no brood) of <i>drh-3(sam27)</i> at permissive temperature. C) STYO14 staining of dissected germlines shows RNA pooling around the nuclear periphery and in the rachis of both <i>csr-1</i> and <i>elli-1</i> mutants. D) Model for CSR-1 and ELLI-1 function. P granules (yellow) reside on the nuclear periphery where they receive transcripts coming through the nuclear pore complex. The CSR-1 complex functions in P granules to recognize germline abundant or licensed transcripts. Our model is that CSR-1 and ELLI-1 function to move germline abundant or licensed transcripts through and away from P granules and into the cytoplasm. In mutants this dispersal of RNA is blocked, RNA and P granules accumulate, and fertility and RNAi efficacy is decreased. It remains unclear whether ELLI-1 directly interacts with RNA.</p

<i>elli-1</i> alleles and phenotypes.

<p>A) Location and flanking sequences of <i>elli-1</i> alleles. Red lines show early stop codons, grey lines show base pair substitutions and deletions. Yellow boxes indicate a codon in the reading frame. ELLI-1’s N-terminal predicted disordered region is shown by the green bars. B) <i>elli-1(sam3)</i> and <i>elli-1(RNAi)</i> cause bright expression of enlarged PGL-1::GFP granules. C) Fosmid rescue of bright PGL-1::GFP expression in <i>elli-1(sam3)</i> animals. Red body-wall muscles mark animals with the rescuing fosmid. D) Cross-section through pachytene germ cells on the surface of wild-type and <i>elli-1(sam3)</i> germlines stained with anti-PGL-1 (green), anti-Nuclear Pore Complex antibody mAb414 (red), and DAPI/DNA (blue). P granules in <i>csr-1</i> and <i>elli-1</i> germlines are enlarged and often detach from the nuclear periphery. E) Cross-section through <i>wild-type</i> and <i>elli-1</i> germlines with the same strains as in D. Arrows show PGL-1 accumulation in the shared cytoplasm. F) GFP tagging endogenous GLH-1 in germlines of <i>wild-type</i> and <i>elli-1(sam3)</i> worms.</p