We further examined worms recovering from 4 d of L1 starvation and found that around 90% of the mir-71(lf) mutants displayed retarded vulval precursor cell (VPC) division, compared with less than 5% in wild type (Fig. 4A). In contrast, the nuclear-localized GFP expression under the control of the 3′UTR of age-1(Fig. 3 C and D) or unc-31 (Fig. 3 E and F) was strongly repressed in the control worms, but prominently derepressed in mir-71(lf) mutant worms. If the 3′UTR of age-1 or unc-31 is repressed by miR-71, the GFP expression will be repressed in tissues where miR-71 is expressed in wild-type worms, but derepressed in the same tissues of mir-71(lf) worms. Among short-lived miRNA mutants, a mir-71 deletion mutant, mir-71(n4115) (referred to as mir-71(lf) hereafter), displayed a severe reduction in L1 starvation survival rate (Table S1 and Fig. 2A).
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- If this were true, the starved mir-71(lf); daf-16(lf) double-mutant worms should show a slow growth phenotype similar to that of daf-16(lf) worms, but no specific VPC timing defect.
- Mutating miR-71 drastically reduces the survival rate of animals in L1 diapause, and the effect can be suppressed by mutations of insulin receptor pathway genes age-1 and unc-31.
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- Because miRNA-mediated gene silencing may cause translational inhibition or mRNA degradation or both (19), the relatively small increase of UNC-31 in mir-71(lf) animals was still consistent with unc-31 being a target of miR-71.
Cell cycle follows “pause and play” mechanism in environment stress recovery in diverse plant species
The reporter is strongly expressed in H and V cells in both wild-type and mir-71(lf) worms. Because miRNA-mediated gene silencing may cause translational inhibition or mRNA degradation or both (19), the relatively small increase of UNC-31 in mir-71(lf) animals was still consistent with unc-31 being a target of miR-71. To test whether the activity of the InsR pathway was down-regulated by miR-71, we first examined the endogenous expression of components of the InsR pathway in mir-71(lf). (C) The poor survival rate of daf-16(mu86, null) was enhanced by mir-71(lf).
REDIRECTING TO HACKED ACCOUNT RECOVERY
MiR-71 regulates vulval cell division during recovery of starved L1 worms. We thus asked whether miR-71 was required for the reinitiation of developmental programs during the recovery phase after L1 starvation. These results indicate that miR-71 is not essential for arresting seam cell or M-cell divisions during L1 diapause, suggesting that miR-71 function is distinct from DAF-16 function. DAF-16 (the FOXO homolog in C. elegans) has been shown to play an important role in cell cycle arrest and developmental progression partly by promoting cki-1 expression in some somatic cells during L1 arrest (2). S1A indicated a dominant role of intestinal miRNAs in regulating L1 starvation survival.
We further found that this survival rate reduction of ain-1 mutants was overcome by ectopic expression of the AIN-2 protein in the intestine but not in the muscle (Fig. 1A and Fig. S1A). In this study, we addressed the questions of whether and how miRNAs impact developmental arrest and the long-term survival of early L1 stage worms in response to food starvation. Furthermore, a recent study suggests that the expression of certain miRNAs is differentially regulated by starvation-induced dauer diapause (15). Consistent with these ideas, several recent lines of evidence suggest that miRNA let-7 and the heterochronic genes lin-42 and hbl-1 are required to regulate the starvation-induced dauer diapause (10–12) and that a number of miRNAs including lin-4 and mir-71 are involved in regulating life span (13, 14).
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- (H) Fluorescence and DIC images showing that a lin-42 3′UTR reporter was repressed in mir-71(+) worms (2/2 transgenic lines) and prominently derepressed in mir-71(−) worms (2/2 transgenic lines).
- Individual GFP reporter constructs for candidate genes (4 ng/μL) and the mCherry internal control plasmid (4 ng/μL) were mixed with unc-119 rescuing plasmid (20 ng/μL) and pBluescript KS+ (72 ng/μL) and coinjected into unc-119(ed3) and mir-71(n4115); unc-119(ed3) worms following standard protocols (32).
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Furthermore, miR-71 plays a prominent role in developmental recovery from L1 diapause partly through repressing the expression of certain heterochronic genes. Here we show that compromising overall microRNA (miRNA) functions or mutating certain individual miRNAs impairs the long-term survival of nematodes during starvation-induced L1 diapause. But offering people opportunities to explore games and play in a safe, non-abusive, self-directed context like a library seems to me to matter, and for folks who are more likely than most to have social anxieties, finding those opportunities can be extremely hard. This certainly accords with the rise of therapists using games as therapy – a web search for “Dungeons & Dragons therapy” shows the recent explosion in ways in which people are finding healing in play. That means that when sales are up, more people are playing this and other older, less-fun games instead of games with the advantage of more than a century’s advancements in the art and revery play science of play, including co-operative games, social games, nomic games, and more. The PlayStation Family app lets you manage your family from your mobile device, and offers additional options to monitor your children’s play time.
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There may be one (or more) that are uncomfortable or downright piss you off. Stacking together concepts, ideas, and ways of coping that don’t include alcohol as a way to navigate to a better life and get closer to our true selves. There are as many ways to live free of drugs and alcohol as there are stories of successful sober people. Create your account and connect with a world of communities.
Waking up not knowing what you did last night and that when we promise we’ll never drink again, it’s quite fine when we reach for the wine and wine glass the next weekend when happy hour hits. And for those of us who have taken a step into the other side, regardless of sobriety time, see it. A mechanism through which we can move through our addiction and keep saying YES to an AF life.
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We identified a conserved “pause and play” mechanism of cell cycle during environmental stress and recovery. On 31 October, the band announced the 2025–2026 Tanzneid World Tour and released the song „Tanzneid“ with a music video featuring Zummo on drums for the first time. On 29 April, Friedrich announced his departure from Electric Callboy after 13 years with the band, stating his intention to pursue a new direction in life. In March 2022, they announced that they had changed their name from Eskimo Callboy to Electric Callboy after learning that „Eskimo“ can be seen as an outdated term and slur for the Inuit and Yupik people.
Writing it out, playing both tapes forward, as plain as day on paper, did wonderful things. One who loves spending time with their family during the holidays and cooking delicious meals, despite the stress it sometimes brings on. The space for me to, for the first time in my adult life, truly decide how I want to live.
