A Single Gene Target of an ETS-Family Transcription Factor Determines Neuronal CO2-Chemosensitivity

Many animals possess neurons specialized for the detection of carbon dioxide (CO2), which acts as a cue to elicit behavioral responses and is also an internally generated product of respiration that regulates animal physiology. In many organisms how such neurons detect CO2 is poorly understood. We report here a mechanism that endows C. elegans neurons with the ability to detect CO2. The ETS-5 transcription factor is necessary for the specification of CO2-sensing BAG neurons. Expression of a single ETS-5 target gene, gcy-9, which encodes a receptor-type guanylate cyclase, is sufficient to bypass a requirement for ets-5 in CO2-detection and transforms neurons into CO2-sensing neurons. Because ETS-5 and GCY-9 are members of gene families that are conserved between nematodes and vertebrates, a similar mechanism might act in the specification of CO2-sensing neurons in other phyla

Fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin with gemtuzumab ozogamicin improves event-free survival in younger patients with newly diagnosed aml and overall survival in patients with npm1 and flt3 mutations

Purpose To determine the optimal induction chemotherapy regimen for younger adults with newly diagnosed AML without known adverse risk cytogenetics. Patients and Methods One thousand thirty-three patients were randomly assigned to intensified (fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin [FLAG-Ida]) or standard (daunorubicin and Ara-C [DA]) induction chemotherapy, with one or two doses of gemtuzumab ozogamicin (GO). The primary end point was overall survival (OS). Results There was no difference in remission rate after two courses between FLAG-Ida + GO and DA + GO (complete remission [CR] + CR with incomplete hematologic recovery 93% v 91%) or in day 60 mortality (4.3% v 4.6%). There was no difference in OS (66% v 63%; P = .41); however, the risk of relapse was lower with FLAG-Ida + GO (24% v 41%; P < .001) and 3-year event-free survival was higher (57% v 45%; P < .001). In patients with an NPM1 mutation (30%), 3-year OS was significantly higher with FLAG-Ida + GO (82% v 64%; P = .005). NPM1 measurable residual disease (MRD) clearance was also greater, with 88% versus 77% becoming MRD-negative in peripheral blood after cycle 2 (P = .02). Three-year OS was also higher in patients with a FLT3 mutation (64% v 54%; P = .047). Fewer transplants were performed in patients receiving FLAG-Ida + GO (238 v 278; P = .02). There was no difference in outcome according to the number of GO doses, although NPM1 MRD clearance was higher with two doses in the DA arm. Patients with core binding factor AML treated with DA and one dose of GO had a 3-year OS of 96% with no survival benefit from FLAG-Ida + GO. Conclusion Overall, FLAG-Ida + GO significantly reduced relapse without improving OS. However, exploratory analyses show that patients with NPM1 and FLT3 mutations had substantial improvements in OS. By contrast, in patients with core binding factor AML, outcomes were excellent with DA + GO with no FLAG-Ida benefit

COVID-19 Pandemic Impact on Percutaneous Coronary Intervention for Acute Coronary Syndromes: An Australian Tertiary Centre Experience

BACKGROUND: Countries who suffered large COVID-19 outbreaks reported a decrease in acute coronary syndrome (ACS) presentations and percutaneous coronary intervention (PCI). The impact of the pandemic in countries like Australia, with relatively small outbreaks yet significant social restrictions, is relatively unknown. There is also limited and conflicting data regarding the impact on clinical outcomes, symptom-to-door time (STDT) and door-to-balloon time (DTBT). METHODS: Consecutive ACS patients treated with PCI were prospectively recruited from a tertiary hospital network in Melbourne, Australia. The pre-pandemic period (11 March 2019-10 March 2020) was compared to the pandemic period (11 March 2020-10 May 2020) using an interrupted time series analysis with a primary endpoint of number PCI-treated ACS per day. Secondary endpoints included STDT, DTBT, total mortality and major adverse cardiac events (MACE). RESULTS: A total 984 ACS patients (14.8% during the pandemic period) received PCI. Mean number of PCI-treated ACS per day did not differ between the two periods (2.3 vs 2.4, p=0.61) with no difference in STDT [+51.3 mins, 95% confidence interval (CI) -52.4 to 154.9, p=0.33], 30-day mortality (5% vs 5.3%, p=0.86) or MACE (5.2% vs 6.1%, p=0.68). DTBT was significantly longer during the pandemic versus the pre-pandemic period (+18.1 mins, 95% CI 1.6-34.5, p=0.03) and improved with time (slope estimate: -0.76, 95% CI -1.62 to 0.10). CONCLUSIONS: Despite significant social restrictions imposed in Melbourne, numbers of ACS treated with PCI and 30-day outcomes were similar to pre-pandemic times. DTBT was significantly longer during the COVID-19 pandemic period, likely reflecting infection control measures, which reassuringly improved with time

Sex disparities in myocardial infarction : Biology or bias?

Women have generally worse outcomes after myocardial infarction (MI) compared to men. The reasons for these disparities are multifactorial. At the beginning is the notion—widespread in the community and health care providers—that women are at low risk for MI. This can impact on primary prevention of cardiovascular disease in women, with lower use of preventative therapies and lifestyle counselling. It can also lead to delays in presentation in the event of an acute MI, both at the patient and health care provider level. This is of particular concern in the case of ST elevation MI (STEMI), where “time is muscle”. Even after first medical contact, women with acute MI experience delays to diagnosis with less timely reperfusion and percutaneous coronary intervention (PCI). Compared to men, women are less likely to undergo invasive diagnostic testing or PCI. After being diagnosed with a STEMI, women receive less guideline-directed medical therapy and potent antiplatelets than men. The consequences of these discrepancies are significant—with higher mortality, major cardiovascular events and bleeding after MI in women compared to men. We review the sex disparities in pathophysiology, risk factors, presentation, diagnosis, treatment, and outcomes for acute MI, to answer the question: are they due to biology or bias, or both

IRK-1 Potassium Channels Mediate Peptidergic Inhibition of Caenorhabditis elegans Serotonin Neurons via a Gₒ Signaling Pathway

To identify molecular mechanisms that function in G-protein signaling, we have performed molecular genetic studies of a simple behavior of the nematode Caenorhabditis elegans, egg laying, which is driven by a pair of serotonergic neurons, the hermaphrodite-specific neurons (HSNs). The activity of the HSNs is regulated by the Gₒ-coupled receptor EGL-6, which mediates inhibition of the HSNs by neuropeptides. We report here that this inhibition requires one of three inwardly rectifying K+ channels encoded by the C. elegans genome: IRK-1. Using ChannelRhodopsin-2-mediated stimulation of HSNs, we observed roles for egl-6 and irk-1 in regulating the excitability of HSNs. Although irk-1 is required for inhibition of HSNs by EGL-6 signaling, we found that other Gₒ signaling pathways that inhibit HSNs involve irk-1 little or not at all. These findings suggest that the neuropeptide receptor EGL-6 regulates the potassium channel IRK-1 via a dedicated pool of Gₒ not involved in other Gₒ-mediated signaling. We conclude that G-protein-coupled receptors that signal through the same G-protein in the same cell might activate distinct effectors and that specific coupling of a G-protein-coupled receptor to its effectors can be determined by factors other than its associated G-proteins.National Institutes of Health (U.S.) (NIH Grant R01-GM024663)National Institutes of Health (U.S.) (NIH Grant R01-GM098320

Comparison of late cardiac death and myocardial infarction rates in women vs men with ST-elevation myocardial infarction

Women and patients with incomplete revascularization (IR) have a worse prognosis after ST elevation myocardial infarction (STEMI). However, the extent to which IR affects outcomes for women with STEMI compared with men is not well characterized. Thus, we examined late outcomes of 589 consecutive STEMI patients who received percutaneous coronary intervention and assessed SYNTAX scores (SS), both at baseline and after all procedures (residual SS). A residual SS >8 defined IR. The primary end point was cardiac death or myocardial infarction (MI), with median follow-up of 3.6 years [interquartile range [IQR] 2.6 to 4.7]. Women (n = 123) had lower baseline SSs 15.0 [IQR 9 to 20], than men (n = 466), 16.0 [IQR 9 to 20; p = 0.02. After all planned procedures, the residual SS was 5.0 [IQR 0 to 9] in women and 5.0 (IQR 1 to 11] in men, p = 0.37. Cardiac death or MI occurred in (97/589) patients (16%), 24% (30/123) in women and 14% (67/466) in men (hazard ratio [HR] 1.75; 95% confidence intervals [CI] 1.14 to 2.69; p = 0.01). In patients with residual SYNTAX score (rSS) >8 cardiac death or MI occurred in 43% (15/35) of women and 23% 36/158 men (HR 2.14; 95% CI 1.17 to 3.91; p = 0.01). In patients with rSS = 0 to 8 cardiac death or MI occurred in 17% (15/88) of women and 10% of men (31/308) (HR 1.68; 95% CI 0.91 to 3.12; p = 0.10; interaction p value 0.58). Multivariate analysis found women were 1.77 times more likely than men to experience cardiac death or MI (95% CI 1.13 to 2.77; p = 0.01). In conclusion, we found despite a lower burden of disease at presentation and no difference in rates of IR between men and women, outcome differences were substantial. Women with rSS >8 were twice as likely as men with the same rSS to experience cardiac death or MI post-STEMI. Differences remained significant postrisk adjustment

Sex Differences in Outcome and Prescribing Practice in ST-elevation MI Patients with Multivessel Disease and Incomplete Revascularisation

Objective: To investigate the extent to which multivessel disease, incomplete revascularisation and prescribing differences contribute to sex-based outcome disparities in patients with ST-elevation MI (STEMI) and establish whether differences in cardiac death and MI (CDMI) rates persist at long-term follow-up. Methods and results: This observational study evaluates sex-based outcome differences (median follow-up 3.6 years; IQR [2.4–5.4]) in a consecutive cohort of patients (n=2,083) presenting with STEMI undergoing percutaneous coronary intervention). Of the studied patients 20.3% (423/2,083) were women and 38.3% (810/2,083) had multivessel disease (MVD). Incomplete revascularisation was common. The median residual SYNTAX score (rSS) was 5.0 (IQR [0–9]) in women and 5.0 (IQR [1–11]) in men (p=0.369), and in patients with MVD it was 9 (IQR [6–17]) in women and 10 (IQR [6–15]) in men (p=0.838). The primary endpoint CDMI occurred in 20.3% of women (86/423) and in 13.2% of men (219/1,660) (p=0.028). Differences persisted following multivariable risk adjustment: female sex was independently associated with CDMI (aHR 1.33; IQR [1.02–1.74]). Women with MVD had CDMI more often than all other groups (p8. Observed differences in P2Y12 prescribing practices may contribute to poor outcomes for women with MVD and incomplete revascularisation

Expression of the ETS-5 target gene <i>gcy-9</i> restores CO₂-chemosensitivity to <i>ets-5</i> mutants.

<p>(A) <i>gcy-36</i> and <i>gcy-18</i> promoters, which are active in oxygen-sensing and thermosensory neurons, respectively, are not regulated by ETS-5. Shown are lateral views of wild-type and <i>ets-5</i> mutant animals carrying either a <i>gcy-36</i> reporter, which is expressed by the oxygen-sensing URX neurons, or a <i>gcy-18</i> reporter, which is expressed by the thermosensory AFD neurons. A: anterior, V: ventral, scale bar is 20 µm. The <i>Prom_gcy-36::cameleon</i> transgene used was <i>wzEx39</i> and the <i>Prom_gcy-18::gfp</i> transgene was <i>wzEx40</i>. (B) Expression of <i>gcy-9</i> in either the URX oxygen sensors or the AFD thermosensors rescues the behavioral defect of <i>ets-5</i> mutants. The left plot shows the mean fraction of animals ± SEM that reversed during a four second exposure to either control atmosphere (0% CO₂, 20% O₂, balance N₂) or CO₂-enriched atmosphere (10% CO₂, 20% O₂, balance N₂). The right plot shows the effect of CO₂ on reversals as measured with an avoidance index, as in Fig. 2C. Plotted are the mean avoidance indices for each of the four strains tested ± SEM. P values were calculated by one-way ANOVA. <i>N</i> = 3–5 populations of 30–50 animals. The <i>ets-</i>5 mutant strain used was FX1734. The <i>Prom</i>_{gcy<i>-36</i>}<i>::gcy-9</i> transgene used was <i>wzIs97</i> and the <i>Prom_gcy-18::gcy-9</i> transgene was <i>wzEx34</i>. (C) Expression of <i>gcy-9</i> in the URX oxygen sensors confers sensitivity to CO₂. The ratiometric calcium indicator cameleon was expressed in the URX neurons. Wild-type URX neurons (top panel) showed a small decrease in the ratio of YFP:CFP emissions in response to 10 s CO₂ pulses indicating decreases in cell calcium. URX neurons expressing <i>gcy-9</i> showed increases in cell calcium in response to CO₂ stimuli, with an average ratio change of greater than 20% (lower panel). The cameleon expression transgene used was <i>wzIs96[Prom_gcy-32::cameleon]</i> and the <i>gcy-9</i> expression transgene was <i>wzIs97[Prom_gcy-36::gcy-9].</i> (D) Expression of <i>gcy-9</i> in the AFD thermosensors confers sensitivity to CO₂. Calcium responses of wild-type AFD neurons (top panel) and AFD neurons that express <i>gcy-9</i> (bottom panel) in responses to a 10% CO₂ stimulus. The cameleon expression transgene was <i>fxIs105[Prom_gcy-8::cameleon]</i>, and the <i>Prom_gcy-18::gcy-9</i> transgene was <i>wzEx34.</i> For panels C and D, plots are mean YFP/CFP emissions ratios normalized to the pre-stimulus ratio R₀ (<i>N</i> = 16–22 animals). Red shaded areas represent SEM.</p

An ETS-family transcription factor is required for the specification of <i>C. elegans</i> CO₂-chemosensitive BAG neurons.

<p>(A) A 31 basepair DNA element comprising a single ETS-binding motif (top) drives expression of GFP specifically in the BAG chemosensitive neurons (bottom). (B) One of ten ETS-family transcription factors encoded by the <i>C. elegans</i> genome is required for specification of BAG neurons. Shown is percent of animals mutant for each of ten ETS-family transcription factors encoded by the <i>C. elegans</i> genome that are BAGL/R ON (green circles) and BAGL/R OFF (open circles) for expression of a <i>Prom_flp-19::gfp</i> reporter transgene. <i>N</i> = number of animals scored. # We found one <i>lin-1(e1777)</i> mutant in which <i>Prom_flp-19::gfp</i> was not expressed in BAGR. (C) Fluorescence micrographs of <i>Prom_flp-19::gfp</i> expression in a wild-type animal, an <i>ets-5</i> mutant and an <i>ets-5</i> mutant carrying a wild-type copy of the <i>ets-5</i> locus in a fosmid-derived transgene. BAGL/R neuron positions are marked by red circles and cells previously identified as AWAL/R <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034014#pone.0034014-Kim1" target="_blank">[19]</a> are marked by blue circles. The nerve ring is indicated by an arrowhead. The scale bar in lower panel is 20 µm. A: anterior, L: left. The <i>ets-5</i> mutant allele was <i>tm1734</i>. The <i>Prom_flp-19::gfp</i> transgene was <i>ynIs34</i> and the <i>ets-5</i> rescuing transgene was <i>rpEx246</i>.</p

ETS-5 directly interacts with the <i>gcy-9</i> promoter.

<p>(A) ETS-5::GFP associates with the <i>gcy-9</i> promoter <i>in vivo</i>. Anti-GFP immunoprecipitates were prepared from cross-linked extracts of wild-type animals or animals carrying a functional <i>ets-5::gfp</i> transgene and interrogated for the presence of <i>gcy-9</i> promoter sequences by PCR. Immunoprecipitates from transgenic animals were enriched for <i>gcy-9</i> promoter sequences that contained the ETS-binding site at −202 bp. Control sequences at −5000 bp were not enriched in immunoprecipitates from transgenic animals. The <i>ets-5::gfp</i> transgene used was <i>wzIs80</i>. (B) ETS-5 binds to <i>gcy-9</i> promoter sequences <i>in vitro</i>. A mobility shift assay was performed with recombinant GST::ETS-5 and a 45 bp biotinylated DNA duplex probe containing the ETS-binding site from the <i>gcy-9</i> promoter. Recombinant GST::ETS-5 but not GST alone altered the electrophoretic mobility of the probe. The interaction between GST::ETS-5 and the probe was blocked by a molar excess of unlabeled probe but not by an excess of scrambled probe with the same nucleotide composition. Excess unlabeled wild-type and scrambled competitor probe was added in the following molar ratios: 10×, 50×, 100×, 500×.</p