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).
Given my views on play, you won’t be surprised to learn that I asked the presenters about whether play experiences could qualify as the kinds of responsive, positive, engaged, and empowering interactions they described. I was attending professional development aimed at helping library staff support families from refugee backgrounds, and we were discussing the role that libraries and childcare staff (mostly the latter, based on the mix of the group) can play in helping often highly traumatised communities. A small increase in risk of harm, replicated many times on a massive scale, means harm is certainly occurring. As COVID-19 lockdowns unfolded around the world, and sales of games, puzzles, toys, and other playful artforms spiked, a friend asked me if I felt a sense of validation. And worst of all, a combination of the two, where other players essentially become supporting cast for a scripted victory that is neither earned, nor fun, nor remotely playful – a psychodrama rather than a game, toying with their victims rather than playing with them. Controlling bullies who insist everyone play their way, forcing people to make deals they don’t want and to keep playing even when they aren’t having fun.
- Paul encourages you to check in with yourself about your feelings about your AF journey.
- 1A because the ain-1 mutations reduce, but do not eliminate, miRISC functions.
- 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).
- (C) Fluorescence and differential interference contrast (DIC) images showing that the age-1 3′UTR reporter was repressed in mir-71(+) worms (3/4 transgenic lines) but not in mir-71(lf) worms (4/4 transgenic lines).
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In December, the band announced that they were removing a selection of their old songs from all platforms due to lyrics that had „become increasingly problematic in modern times“. Since she got sober at 19, she has been revisiting fun at her current revery play stage of life. Paul encourages you to check in with yourself about your feelings about your AF journey.
- At Khiron Clinics, we provide group therapy in carefully structured formats designed to support nervous system regulation, emotional resilience, and embodied healing.
- Furthermore, miR-71 plays a prominent role in developmental recovery from L1 diapause partly through repressing the expression of certain heterochronic genes.
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- In fact, one of the great, almost pre-verbal lessons of play is precisely that the magic circle of play exists only with, is in large part comprised of, the willing participation of everyone in and around it.
- 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.
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On 12 February 2020, they announced that lead singer Biesler would be leaving the band. In March, the band toured Europe supported by Iwrestledabearonce, Her Bright Skies, and To the Rats and Wolves. We Are the Mess peaked at No. 8 on Germany’s official long-play charts and No. 64 in Austria.
For younger players
Knocking down lit-1 by RNAi in mir-71(lf); lin-42(lf) double mutants caused no significant suppression of the VPC timing defects of mir-71(lf) worms. To determine the functional relationship of miR-71 with LIN-42 and LIT-1, mir-71(lf); lin-42(lf) L1 worms were starved for 4 d and recovered on lit-1(RNAi) plates. The strong suppression of the mir-71(lf) defect by hbl-1(RNAi), and the relatively weak effect of miR-71 on hbl-1 expression, are consistent with the idea that miR-71 exerts its role by modulating activities of multiple genes related to hbl-1 function in developmental timing. We then compared the expression of a hbl-1 3′UTR reporter (18) in the mir-71(lf) mutants with that in wild type and found that the expression of this reporter was slightly derepressed at L3 in the mir-71 mutant (Fig. 4 F and G).
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It’s also notorious among tabletop enthusiasts for making some people loathe the entire medium of board games… I’ve got several posts to write on this topic, but I want to start by honouring Gene’s remarks with a ringing endorsement of his point about supporting those with trauma responses to play, and building on what that means more broadly. I’d like to thank whoever chose the GLAM Blog Club theme for this month, and highlight some of my past posts as potential inspiration for anyone thinking of writing about play and games in a libraries/GLAM context – specifically my Talking Points series, especially the second one, on the powers of play. We detected that you have an open account recovery request. Penalties are applied no matter who was playing at the time.
Playing The Tape Forward: A Foundation For My Sobriety
(H and I) Fluorescence images (H) and statistical data (I) showing that the M cell diveded in fed animals but remained undivided in 4-, 7-, or 11-d–starved L1 wild-type and mir-71(lf) worms. (E) Fluorescence and DIC images showing that the unc-31 3′UTR reporter was repressed in mir-71(+)worms (2/2 transgenic lines) but not in mir-71(lf) worms (4/4 transgenic lines). We found that the reduced survival rate of ain-1 was suppressed by either reduction of age-1 function or loss of unc-31 function (Fig. 1 B and C), suggesting that a significant portion of the overall miRNA functions in L1 diapause is upstream of, or in parallel to, the InsR pathway. Previous studies indicate that the InsR pathway plays a dominant role in regulating L1 starvation survival and that reducing the activity of the insulin receptor daf-2, the PI3Kinase age-1, or the upstream regulator unc-31 results in increased L1 starvation survival rate (2, 3). The two ain-1 loss-of-function alleles displayed significant reductions in L1 starvation survival rate. We found that ain-1 but not ain-2 mutants displayed a significant reduction in L1 starvation survival rate compared with that of wild type (Fig. 1 A and D).
The overall effect of miRNAs on L1 starvation survival is expected to be significantly stronger than that reflected by the data in Fig. These results suggest that miRNAs act in the intestine, and possibly in other tissues, to promote L1 starvation survival. However, the mechanisms that coordinate the long-term survival, overall developmental arrest, and reinitiation remain to be investigated.
The transcript level of unc-31 was increased in mir-71(lf) worms, compared with that of wild-type controls that were normalized to the value of 1. MiR-71 represses the expression of age-1 and unc-31 through the actions on their 3′UTR, but miR-71 is not required for arresting M cell division during L1 diapause. We found that the poor survival rate of daf-16(mu86)(lf) was further decreased by mir-71(lf) (Fig. 2C), consistent with the notion that a portion of miR-71 activities regulate genes that act in parallel to UNC-31–mediated InsR/PI3K signaling for long-term survival during L1 diapause. (B) The severely reduced survival rate of the mir-71(lf) mutant was suppressed by a null allele of unc-31(e928).
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Elegans Genetic Center for many mutants of miRNA and other genes. Images were pseudocolored in Photoshop CS3 (Adobe) and assembled in Illustrator CS3 (Adobe). The data for 3′UTR expression and for VPC timing were analyzed using χ2 test. To determine viability, 20-μL aliquots (60–100 worms) were placed every 3 d onto two 6-cm nematode growth medium (NGM) plates seeded with OP50, and the numbers of L1 worms were recorded as number of plated worms (Np). A total of 16–24 h later, the density of newly hatched L1 worms was adjusted to three to five worms per microliter S-basal. The eggs were transferred to plates seeded with HB101 and bleached again 3 d later.
This feels like one of those posts where if I had more time I could make it shorter… but I don’t. Fun seems self-evident, but I would just gently remind readers that fun is non-trivial; is in fact the active face of joy, as beauty is the passive. As above, play without consent, treating others as toys without independent will rather than as fellow players, is oppression and abuse. In fact, one of the great, almost pre-verbal lessons of play is precisely that the magic circle of play exists only with, is in large part comprised of, the willing participation of everyone in and around it. That curation isn’t only about having collections available for loan or in the library space; it’s also about making actual play experiences available.
I’m fighting for a reimbursement, but I’m posting this so you don’t lose the resources I did.“ Support isn’t just unhelpful; they aren’t even in the same reality as the players. „To the people calling me an ‚EA Spokesman’—take a look at my latest interaction with them. This study is supported by National Institutes of Health Grant GM47869 (to M.H.).