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A conditional knock-in mouse was generated in which the TAK1 catalytic subunit was largely replaced by the kinase-inactive TAK1[D175A] mutant in immune cells. The activation of p38α MAP kinase, c-Jun N-terminal kinases 1 and 2 (JNK1/2) and the canonical IKK complex induced by stimulation with several TLR-activating ligands was reduced in bone marrow-derived macrophages (BMDM) from TAK1[D175A] mice. TLR signalling in TAK1[D175A] BMDM was catalysed by the residual wild-type TAK1 in these cells because it was abolished by either of two structurally unrelated TAK1 inhibitors (NG25 and 5Z-7-oxozeaenol) whose off-target effects do not overlap. The secretion of inflammatory mediators and production of the mRNAs encoding these cytokines induced by TLR ligation was greatly reduced in peritoneal neutrophils or BMDM from TAK1[D175A] mice. The Pam3CSK4- or LPS-stimulated activation of MAP kinases and the canonical IKK complex, as well as cytokine secretion, was also abolished in TAK1 knock-out human THP1 monocytes or macrophages. The results establish that TAK1 protein kinase activity is required for TLR-dependent signalling and cytokine secretion in myeloid cells from mice. We discuss possible reasons why other investigators, studying myeloid mice with a conditional knock-out of TAK1 or a different conditional kinase-inactive knock-in of TAK1, reported TAK1 to be a negative regulator of LPS-signalling and cytokine production in mouse macrophages and neutrophils.

Frosty pod rot (FPR), caused by Moniliophthora roreri (MR), is one of the most important diseases of cacao, causing losses of up to 90% in areas where it is present. A limited number of resistant clones have been identified, but additional sources of disease resistance are being sought by breeding programs. RNA-sequencing was used to identify differences in constitutive (mock-inoculated) and pathogen-induced gene expression in cacao pods from trees resistant and susceptible to MR. The greatest number of differentially expressed genes (DEGs) (n=2,039) was identified in susceptible material following inoculation with MR (7 days post inoculation), with the most common pathway categories being cell growth/death (n=102), signaling (n=308), and carbohydrate metabolism (n=141). Resistant material showed little overall change in gene expression (5 DEGs) following pathogen inoculation, suggesting that infection had not become established. For this reason, differences in constitutive gene expression are likely to provide the most relevant information on FPR resistance in this study. In the absence of the pathogen, 144 genes were expressed at a higher level in the resistant material, and 265 at a higher level in the susceptible material. The genes most highly upregulated in resistant material encode for disease resistance proteins in the leucine rich repeat family and cysteine-rich receptor-like protein kinases. Ten DEGs were located within QTL on chromosomes 2, 7, and 9 previously associated with FPR resistance in this population. This study provides the first analysis of constitutively expressed genes contributing to FPR resistance and identifies significant genes within known disease resistance QTL.

Thread blight disease (TBD) is a serious emerging threat to global cacao production, causing leaf blight symptoms and necrosis in the canopy of infected trees. Though previously thought to be caused primarily by the fungus Marasmiellus scandens, recent work has revealed that a variety of pathogenic Marasmiaceae are responsible for TBD, with Marasmius tenuissimus identified as the dominant causal agent of TBD in Ghanaian cacao plantations. M. tenuissimus has also been reported in remote organic cacao plantations in the Amazonas Department of Peru, infecting 90-100% of the trees surveyed. Here we have assembled and annotated the nuclear genome of M. tenuissimus isolate GH-37. This resource is a first attempt at a high-quality draft genome utilizing both second- and third-generation sequencing for M. tenuissimus and extends the current understanding of the evolution, ecology, and virulence of this pathogen.

As students, teachers, researchers, and citizens, how can we tell what’s true? For people outside of academic disciplines, peer review is seen as a core quality – part of the definition of science many people learn. It has become part of the social contract between scientists and the public. In our highly-contentious social and political milieu, where researchers are sometimes depicted as just another interest group competing to establish their own self-serving version of truth, peer review sends an important signal that scientists aren't afraid of independent scrutiny of our work. However, a lot of research labeled “peer reviewed” is not a reliable source of true information, especially in cases of deliberate manipulation. The pandemic clearly showed we can’t just choose to live in an idealized world of verified truths. But we should not elevate the formal peer review process to an unrealistically sacred status. The quality of our information is vitally important, and there are no easy solutions to the problem of deciding where to place our trust. Experts have to earn the trust they need to do their work effectively, and one way they do that is by engaging with the public in responsible and respectful ways.

Previous research has employed an inadequate measure of marriage metabolism, but the concept may be useful for understanding the system of marriage. This paper addresses changes in the incidence of marital events in the United States from 2008 to 2021. I offer a measure, the Total Rate of Marital Events (TRME), of the projected lifetime experience of marital transitions (marriage, divorce, and widowhood) for a life table cohort. I find that the TRME declined steeply over this relatively short period: 22 percent for men and 19 percent for women. All three components declined in every age group below 90. I suggest that the slowing churn of the marriage system reflects the diminished social presence of marriage in daily life – if not its declining importance – which coincides with the increasingly selective status of married life. A higher status marriage system is a smaller, slower, and more stable marriage system.

Xanthomonas euvesicatoria (Xe) and X. vesicatoria (Xv) are two economically important causal agents of bacterial spot (BS) of tomato and pepper. Management of BS in the field requires rapid and accurate detection. This work therefore aimed to develop a pipeline to design a simple, fast, and reliable assay for the detection of Xe and Xv by Loop-Mediated Isothermal Amplification (LAMP). A total of 109 publicly available whole genomic sequences of 24 different species of bacterial pathogens were used to design primers that would amplify the DNA of the two target species. Laboratory testing of the assay was performed on pure bacterial cultures and artificially infected plants, and amplification was conducted with both a sophisticated laboratory instrument and a simple mobile platform. The testing of the assay confirmed its specificity with a sensitivity reaching 1 pg μL−1 for both pathogens with an assay duration of 40 minutes on a mobile detection platform. Our diagnostics development pipeline enables the easy and fast design of a reliable detection assay in the genomics age. By validating the pipeline with Xe and Xv pathogens, we have simultaneously developed an assay with high specificity, sensitivity, and speed, which will allow it to be deployed, contributing to successful management of BS.

The E3 ligase HOIL-1 forms ester bonds in vitro between ubiquitin and serine/threonine residues in proteins. Here, we exploit UbiSite technology to identify serine and threonine residues undergoing HOIL-1 catalysed ubiquitylation in macrophages stimulated with R848, an activator of the TLR7/8 heterodimer. We identify Thr12, Thr14, Ser20 and Thr22 of ubiquitin as amino acid residues forming ester bonds with the C-terminal carboxylate of another ubiquitin molecule. This increases from 8 to 12 the number of ubiquitin linkage types that are formed in cells. We also identify Ser175 of IRAK4, Ser136, Thr163 and Ser168 of IRAK2 and Thr141 of MyD88 as further sites of HOIL-1-catalysed ubiquitylation together with lysine residues in these proteins that also undergo R848-dependent ubiquitylation. These findings establish that the ubiquitin chains attached to components of myddosomes are initiated by both ester and isopeptide bonds. Ester bond formation takes place within the proline, serine, threonine-rich (PST) domains of IRAK2 and IRAK4 and the intermediate domain of MyD88. The ubiquitin molecules attached to Lys162, Thr163 and Ser168 of IRAK2 are attached to different IRAK2 molecules.

The E3 ligase HOIL-1 forms ester bonds in vitro between ubiquitin and serine/threonine residues in proteins. Here, we exploit UbiSite technology to identify serine and threonine residues undergoing HOIL-1 catalysed ubiquitylation in macrophages stimulated with R848, an activator of the TLR7/8 heterodimer. We identify Thr12, Thr14, Ser20 and Thr22 of ubiquitin as amino acid residues forming ester bonds with the C-terminal carboxylate of another ubiquitin molecule, increasing from 8 to 12 the different types of ubiquitin dimer formed in cells. We also identify Ser175 of IRAK4, Ser136, Thr163 and Ser168 of IRAK2 and Thr141 of MyD88 as further sites of HOIL-1-catalysed ubiquitylation together with lysine residues in these proteins that also undergo R848-dependent ubiquitylation. These findings establish that the ubiquitin chains attached to components of myddosomes are initiated by both ester and isopeptide bonds. Ester bond formation takes place within the proline, serine, threonine-rich (PST) domains of IRAK2 and IRAK4 and the intermediate domain of MyD88. The ubiquitin molecules attached to Lys162, Thr163 and Ser168 of IRAK2 are attached to different IRAK2 molecules.

SummaryADP-heptose activates the protein kinase ALPK1 triggering TIFA phosphorylation at Thr9, the recruitment of TRAF6 and the subsequent production of inflammatory mediators. Here, we demonstrate that ADP-heptose also stimulates the formation of Lys63- and Met1-linked ubiquitin chains to activate the TAK1 and canonical IKK complexes, respectively. We further show that the E3 ligases TRAF6 and c-IAP1 operate redundantly to generate the Lys63-linked ubiquitin chains required for pathway activation, which we demonstrate are attached to TRAF6, TRAF2 and c-IAP1, and that c-IAP1 is recruited to TIFA by TRAF2. ADP-heptose also induces activation of the kinase TBK1 by a TAK1-independent mechanism, which require TRAF2 and TRAF6. We establish that ALPK1 phosphorylates TIFA directly at Thr177 as well as Thr9 in vitro. Thr177 is located within the TRAF6-binding motif and its mutation to Asp prevents TRAF6 but not TRAF2 binding, indicating a role in restricting ADP-heptose signalling. We conclude that ADP-heptose signalling is controlled by the combined actions of TRAF2/c-IAP1 and TRAF6.

I argue against a perception of normal modernity that is on some kind of track that we can try to get a handle on while we work around distractions like a pandemic, climate change, diverging and growing inequality, the centrifugal tendency of identities to be fragmenting, and policy incoherence. Instead, I suggest doing family demography in light of disasters, inequalities, identities, and policies as messy and coherent, unpredictable features of society – not as bugs in the ointment of modern progress.

Protein kinases regulate nearly all aspects of cell life, and alterations in their expression, or mutations in their genes, cause cancer and other diseases. Here, we review the remarkable progress made over the past 20 years in improving the potency and specificity of small-molecule inhibitors of protein and lipid kinases, resulting in the approval of more than 70 new drugs since imatinib was approved in 2001. These compounds have had a significant impact on the way in which we now treat cancers and non-cancerous conditions. We discuss how the challenge of drug resistance to kinase inhibitors is being met and the future of kinase drug discovery., Twenty years have passed since the first small-molecule protein kinase inhibitor, imatinib, gained FDA approval. Here, Cohen et al. review advances in improving the potency and specificity of small-molecule protein kinase inhibitors and assess approaches to overcome the challenge of drug resistance. Applications of these compounds in cancers and other disorders, as well as future directions in the field, are discussed.

The salt-inducible kinases, SIK1, SIK2 and SIK3, most closely resemble the AMP-activated protein kinase (AMPK) and other AMPK-related kinases, and like these family members they require phosphorylation by LKB1 to be catalytically active. However, unlike other AMPK-related kinases they are phosphorylated by cyclic AMP-dependent protein kinase (PKA), which promotes their binding to 14-3-3 proteins and inactivation. The most well-established substrates of the SIKs are the CREB-regulated transcriptional co-activators (CRTCs), and the Class 2a histone deacetylases (HDAC4/5/7/9). Phosphorylation by SIKs promotes the translocation of CRTCs and Class 2a HDACs to the cytoplasm and their binding to 14-3-3s, preventing them from regulating their nuclear binding partners, the transcription factors CREB and MEF2. This process is reversed by PKA-dependent inactivation of the SIKs leading to dephosphorylation of CRTCs and Class 2a HDACs and their re-entry into the nucleus. Through the reversible regulation of these substrates and others that have not yet been identified, the SIKs regulate many physiological processes ranging from innate immunity, circadian rhythms and bone formation, to skin pigmentation and metabolism. This review summarises current knowledge of the SIKs and the evidence underpinning these findings, and discusses the therapeutic potential of SIK inhibitors for the treatment of disease.

Background The prevalence of burnout and depressive symptoms is high among physician trainees. Research Question What is the burden of burnout and depressive symptoms among fellows training in pulmonary and critical care medicine (PCCM) and what are associated individual fellow, program, and institutional characteristics? Study Design and Methods We conducted a cross-sectional electronic survey of fellows enrolled in pulmonary, PCCM, and critical care medicine training programs in the United States to assess burnout and depressive symptoms. Burnout symptoms were measured using the Maslach Burnout Index two-item measure. The two-item Primary Care Evaluation of Mental Disorders Procedure was used to screen for depressive symptoms. For each of the two outcomes (burnout and depressive symptoms), we constructed three multivariate logistic regression models to assess individual fellow characteristics, program structure, and institutional polices associated with either burnout or depressive symptoms. Results Five hundred two of the 976 fellows who received the survey completed it—including both outcome measures—giving a response rate of 51%. Fifty percent of fellows showed positive results for either burnout or depressive symptoms, with 41% showing positive results for depressive symptoms, 32% showing positive results for burnout, and 23% showing positive results for both. Reporting a coverage system in the case of personal illness or emergency (adjusted OR [aOR], 0.44; 95% CI, 0.26-0.73) and access to mental health services (aOR, 0.14; 95% CI, 0.04-0.47) were associated with lower odds of burnout. Financial concern was associated with higher odds of depressive symptoms (aOR, 1.13; 95% CI, 1.05-1.22). Working more than 70 hours in an average clinical week and the burdens of electronic health record (EHR) documentation were associated with a higher odds of both burnout and depressive symptoms. Interpretation Given the high prevalence of burnout and depressive symptoms among fellows training in PCCM, an urgent need exists to identify solutions that address this public health crisis. Strategies such as providing an easily accessible coverage system, access to mental health resources, reducing EHR burden, addressing work hours, and addressing financial concerns among trainees may help to reduce burnout or depressive symptoms and should be studied further by the graduate medical education community.

In 2016 the National Forum for the Enhancement of Teaching and Learning in Higher Education funded three Irish academic libraries to review, implement and evaluate a new National Profession...

HOIL-1, a component of the linear ubiquitin chain assembly complex (LUBAC), ubiquitylates serine and threonine residues in proteins by esterification. Here, we report that mice expressing an E3 ligase-inactive HOIL-1[C458S] mutant accumulate polyglucosan in brain, heart and other organs, indicating that HOIL-1's E3 ligase activity is essential to prevent these toxic polysaccharide deposits from accumulating. We found that HOIL-1 monoubiquitylates glycogen and α1:4-linked maltoheptaose in vitro and identify the C6 hydroxyl moiety of glucose as the site of ester-linked ubiquitylation. The monoubiquitylation of maltoheptaose was accelerated 100-fold by the interaction of Met1-linked or Lys63-linked ubiquitin oligomers with the RBR domain of HOIL-1. HOIL-1 also transferred pre-formed ubiquitin oligomers to maltoheptaose en bloc, producing polyubiquitylated maltoheptaose in one catalytic step. The Sharpin and HOIP components of LUBAC, but not HOIL-1, bound to unbranched and infrequently branched glucose polymers in vitro, but not to highly branched mammalian glycogen, suggesting a potential function in targeting HOIL-1 to unbranched glucosaccharides in cells. We suggest that monoubiquitylation of unbranched glucosaccharides may initiate their removal from cells, preventing precipitation as polyglucosan.

ObjectiveThe aim of the study was to investigate whether the IRAK1/JAK2/Flt3 inhibitor pacritinib prevents disease development in the lupus-prone ABIN1[D485N] knock-in mouse.MethodsABIN1[D485N] knock-in mice aged 8 weeks were fed for 10 weeks on a diet containing pacritinib. Body weight was monitored, and serum collected at the end to measure pacritinib, autoantibody and immunoglobulin levels. Splenic immune cell populations were analysed, and the kidney, liver and lungs examined for pathology.ResultsPacritinib prevented multiple facets of the lupus phenotype in ABIN1[D485N] knock-in mice, including splenomegaly, expansion of splenic germinal centre B cells, follicular T helper cells, and neutrophils, elevated serum levels of double-stranded DNA antibodies and immunoglobulins, glomerular IgA and lung inflammation.ConclusionsPacritinib may be useful for the treatment of multiorgan inflammation in patients with lupus.

Background: Wearing high-heeled shoes is associated with injury risk. During the COVID-19 pandemic, changes in work and social behavior may have reduced women’s use of such footwear. The aim of the study was to investigate how the trend in high-heeled shoe related injuries (HHSRIs) among U.S. women may have been affected by the COVID-19 pandemic. Methods: This used 2016-2020 data from the U.S. Consumer Product Safety Commission’s National Electronic Injury Surveillance System (NEISS) to assess HHSRIs among U.S. women. The study included injuries with the “Footwear” product code and selected those with narratives using words suggesting the involvement of high heels. Analysis is based on frequency of such reports by date. Results: In 2020 there were an estimated 6,290 high-heel related emergency department visits involving women ages 15-69, down from 16,000 per year in 2016-2019. There were 5.40 HHSRIs per 100,000 women ages 15-69 in 2020 (CI: 3.95 to 6.86), significantly below the peak in 2017. Analysis by date shows the 2020 decline began after the start of the COVID-19 shutdowns on March 15. There was no significant change in the percentage of fractures or hospital admissions. Conclusion: The COVID-19 pandemic was associated with a decline in reported injuries related to high-heeled shoes among US women. Among these, there was a non-significant increase in more serious injuries, which might suggest pressure to stay away from hospitals for less serious injuries during the pandemic. However, if fewer women wearing such shoes, the result may be fewer injuries in the future.

Background. Protective facemasks are important for preventing the spread of COVID-19, and almost all Americans have worn them at least some of the time during the pandemic. There are reasonable concerns about some ill effects of mask-wearing, especially for people who wear masks for extended periods, and for the risk of falling as a result of visual obstruction. But there are also unsupported fears and objections stemming from misinformation and fueled by political disputes. Methods. The study analyzed the Consumer Product Safety Commission’s National Electronic Injury Surveillance System (NEISS) for 2020, using the product code for Respiratory Protection Devices, and calculated population incidence rates using Census data. Results. The NEISS included 128 cases, representing an estimated 5122 reported injuries in the population (95% CI 3322, 6922). The overall rate of injury reports was 1.54 per 100,000 U.S. residents (95% CI 1.00, 2.08). People over age 75 had higher rates than the population overall, with 5.27 injuries per 100,000 (95% CI 2.17, 8.37). The most common type of incidents involved facial injuries, rashes, falls, and those that might be considered anxiety-related. Conclusion. Wearing protective face masks is extremely safe, especially in comparison with other common household products, and in light of their protective benefits with regard to prevent the spread of COVID-19. This information may be useful for public health messaging, and for practitioners trying to increase compliance with mask-wearing guidance.

Idiopathic pulmonary fibrosis is a progressive and normally fatal disease with limited treatment options. The tyrosine kinase inhibitor nintedanib has recently been approved for the treatment of idiopathic pulmonary fibrosis, and its effectiveness has been linked to its ability to inhibit a number of receptor tyrosine kinases including the platelet-derived growth factor, vascular endothelial growth factor, and fibroblast growth factor receptors. We show here that nintedanib also inhibits salt-inducible kinase 2 (SIK2), with a similar IC

SUMMARYThe sleep and circadian systems act in concert to regulate sleep-wake timing, yet the molecular mechanisms that underpin their interaction to induce sleep remain unknown. Synaptic protein phosphorylation, driven by the kinase SIK3, correlates with sleep pressure, however it is unclear whether these phosphoproteome changes are causally responsible for inducing sleep. Here we show that the light-dependent activity of SIK1 controls the phosphorylation of a subset of the brain phosphoproteome to induce sleep in a manner that is independent of sleep pressure. By uncoupling phosphorylation and sleep induction from sleep pressure, we establish that synaptic protein phosphorylation provides a causal mechanism for the induction of sleep under different environmental contexts. Furthermore, we propose a framework that details how the salt-inducible kinases regulate the synaptic phosphoproteome to integrate exogenous and endogenous stimuli, thereby providing the molecular basis upon which the sleep and circadian systems interact to control the sleep-wake cycle.

HOIL-1, a component of the Linear Ubiquitin Assembly Complex (LUBAC), ubiquitylates serine and threonine residues in proteins, forming ester bonds (Kelsall et al, 2019, PNAS 116, 13293-13298). Here we report that mice expressing the E3 ligase-inactive HOIL-1[C458S] mutant accumulate polyglucosan in brain, cardiac muscle and other organs, indicating that HOIL-1’s E3 ligase activity is essential to prevent these toxic polysaccharide deposits from accumulating. We found that HOIL-1 monoubiquitylates glycogen and α1:4-linked maltoheptaose in vitro and identify the C6 hydroxyl moiety of glucose as the site of ester-linked ubiquitylation. The HOIL-1-catalysed monoubiquitylation of maltoheptaose was accelerated >100-fold by Met1-linked or Lys63-linked ubiquitin oligomers, which interact with the catalytic RBR domain of HOIL-1. HOIL-1 also transferred preformed ubiquitin oligomers to maltoheptaose en bloc, producing polyubiquitylated maltoheptaose in one catalytic step. The Sharpin and HOIP components of LUBAC, but not HOIL-1, bound to amylose resin in vitro, suggesting a potential function in targeting HOIL-1 to unbranched glucosaccharides in cells. We suggest that monoubiquitylation of unbranched glucosaccharides may initiate their removal by glycophagy to prevent precipitation as polyglucosan.

The rapid formation and activation of the NLRP3 inflammasome is induced by co‐stimulation with LPS and nigericin. It requires the LPS‐stimulated activation of IKKβ, which exerts its effects independently of de novo gene transcription, protein translation and other protein kinases activated by IKKβ. IKKβ is not required for the nigericin‐induced dispersion of the trans‐Golgi network (TGN), but to bring NLRP3 in proximity with TGN38. The nigericin‐induced dispersion of the Golgi is enhanced by co‐stimulation with LPS, and this enhancement is IKKβ‐dependent. Prolonged stimulation with LPS to increase the expression of NLRP3, followed by stimulation with nigericin, produced larger TGN38‐positive puncta, and the ensuing activation of the NLRP3 inflammasome was also suppressed by IKKβ inhibitors added prior to stimulation with nigericin. IKKβ therefore has a key role in recruiting NLRP3 to the dispersed TGN, leading to the formation and activation of the NLRP3 inflammasome., Formation of the NLRP3 inflammasome requires LPS to activate IKKβ and nigericin to disperse the trans‐Golgi network. IKKβ functions to recruit NLRP3 to the dispersed TGN promoting inflammasome formation and activation.

This study aims to measure diversity in scholarly journals’ editorial board structure and characterize patterns of editorial diversity across types of journals. To accomplish these aims, we integrate multiple sources of data at the journal and editor level to assemble a novel database describing the composition of editors and editorial boards for more than six thousand journals internationally, characterized by discipline, commercial publishing model, and research transparency. We then apply name-based gender imputation, geo-entity extraction analysis, and standardized dispersion measures to evaluate each group’s diversity. This analysis reveals that editorial leadership is more homogenous than editorial boards, and that diversity across both boards and leadership varies substantially (and robustly) across disciplines. Open-access journal’s boards exhibit less gender diversity and more international diversity than their closed-access counterparts. These results also suggest that open access, open science, and diversity, and equity, and inclusion are not strongly correlated and thus require separate measurements.

Lung and bladder cancers are mostly incurable because of the early development of drug resistance and metastatic dissemination. Hence, improved therapies that tackle these two processes are urgently needed to improve clinical outcome. We have identified RSK4 as a promoter of drug resistance and metastasis in lung and bladder cancer cells. Silencing this kinase, through either RNA interference or CRISPR, sensitized tumor cells to chemotherapy and hindered metastasis in vitro and in vivo in a tail vein injection model. Drug screening revealed several floxacin antibiotics as potent RSK4 activation inhibitors, and trovafloxacin reproduced all effects of RSK4 silencing in vitro and in/ex vivo using lung cancer xenograft and genetically engineered mouse models and bladder tumor explants. Through x-ray structure determination and Markov transient and Deuterium exchange analyses, we identified the allosteric binding site and revealed how this compound blocks RSK4 kinase activation through binding to an allosteric site and mimicking a kinase autoinhibitory mechanism involving the RSK4's hydrophobic motif. Last, we show that patients undergoing chemotherapy and adhering to prophylactic levofloxacin in the large placebo-controlled randomized phase 3 SIGNIFICANT trial had significantly increased (P = 0.048) long-term overall survival times. Hence, we suggest that RSK4 inhibition may represent an effective therapeutic strategy for treating lung and bladder cancer.

Significance The formation of isopeptide bonds between the C-terminal carboxylate of ubiquitin and ε-amino groups of lysine residues on another protein is a major mechanism for regulating protein function. Ubiquitin can also form peptide bonds with the N-terminal α-amino group of another ubiquitin, a reaction catalysed by the HOIP component of the linear ubiquitin assembly complex (LUBAC). Here, we identify the HOIL-1 component of LUBAC as an unusual ligase that catalyses the formation of oxyester bonds between the C-terminal carboxylate of ubiquitin and serine and threonine residues in other proteins. We identify components of the Myddosome as physiological substrates of HOIL-1, indicating a role for HOIL-1 in regulating innate immunity., The linear ubiquitin assembly complex (LUBAC) comprises 3 components: HOIP, HOIL-1, and Sharpin, of which HOIP and HOIL-1 are both members of the RBR subfamily of E3 ubiquitin ligases. HOIP catalyses the formation of Met1-linked ubiquitin oligomers (also called linear ubiquitin), but the function of the E3 ligase activity of HOIL-1 is unknown. Here, we report that HOIL-1 is an atypical E3 ligase that forms oxyester bonds between the C terminus of ubiquitin and serine and threonine residues in its substrates. Exploiting the sensitivity of HOIL-1–generated oxyester bonds to cleavage by hydroxylamine, and macrophages from knock-in mice expressing the E3 ligase-inactive HOIL-1[C458S] mutant, we identify IRAK1, IRAK2, and MyD88 as physiological substrates of the HOIL-1 E3 ligase during Toll-like receptor signaling. HOIL-1 is a monoubiquitylating E3 ubiquitin ligase that initiates the de novo synthesis of polyubiquitin chains that are attached to these proteins in macrophages. HOIL-1 also catalyses its own monoubiquitylation in cells and most probably the monoubiquitylation of Sharpin, in which ubiquitin is also attached by an oxyester bond. Our study establishes that oxyester-linked ubiquitylation is used as an intracellular signaling mechanism.

Halb-und-Halb-Nägel wurden ursprünglich als transversale Leukonychie bei Patienten mit chronischer Nierenerkrankung beschrieben; später wurden auch andere Erkrankungen (z.B. Morbus Behçet und Morbus Crohn) und Medikamente (wie Isoniazid) mit diesen Veränderungen der Nägel in Verbindung gebracht. Es liegen allerdings auch Berichte über longitudinale Halb-und-Halb-Nägel (mit Veränderung des medialen Anteils der Nägel) an den Großzehen einer älteren Frau sowie, wie nachstehend vorgestellt, an den Daumen eines älteren Mannes vor, die beide weder unter Morbus Behçet, Morbus Crohn oder einer Nierenerkrankung litten noch Isoniazid anwandten. Bei der Frau lagen eine echte Leukonychie, bilaterale Hallux-valgus-Deformität und chronisches Trauma der medialen Nagelfalze vor. Der Mann hatte eine nicht traumatische apparente Leukonychie und bilaterale Koilonychie; der nicht weiße laterale Anteil seines linken Daumennagels wies auch eine lineare Rille (infolge einer digitalen mukoiden Zyste an der proximalen Nagelfalz) und subunguale Hyperkeratose auf. Zusammenfassend lässt sich sagen, dass longitudinale Halb-und-Halb-Nägel ein seltenes Phänomen sind, das bisher mit keiner systemischen Erkrankung oder Medikation in Verbindung gebracht worden ist. Die erworbene Nagelveränderung kann sich entweder als idiopathischer Befund oder nach chronischem Trauma manifestieren.

The atypical E3 ligase HOIL-1 forms ester bonds between ubiquitin and serine/threonine residues in proteins, but the physiological roles of this unusual modification are unknown. We now report that IL-18 signalling leading to the production of interferon γ (IFNγ) and granulocyte-macrophage colony-stimulating factor (GM-CSF) is enhanced in cytotoxic T cells from knock-in mice expressing the E3 ligase-inactive HOIL-1[C458S] mutant, demonstrating that the formation of HOIL-1-catalysed ester-linked ubiquitin bonds restricts the activation of this pathway. We show that the interaction of IRAK2 with TRAF6 is required for IL-18-stimulated IFN-γ and GM-CSF production, and that the increased production of these cytokines in cytotoxic T cells from HOIL-1[C458S] mice correlates with an increase in both the number and size of the Lys63/Met1-linked hybrid ubiquitin chains attached to IRAK2 in these cells. In contrast, the secretion of IL-12 and IL-6 and the formation of il-12 and il-6 mRNA induced in bone marrow-derived macrophages (BMDMs) by prolonged stimulation with TLR-activating ligands that signal via myddosomes, which also requires the interaction of IRAK2 with TRAF6, were not increased but modestly reduced in HOIL-1[C458S] BMDM. The decreased production of these cytokines correlated with reduced ubiquitylation of IRAK2. Our results establish that changes in HOIL-1-catalysed ester-linked ubiquitylation can promote or reduce cytokine production depending on the ligand, receptor and immune cell and may be explained by differences in the ubiquitylation of IRAK2.

The United States experienced a 3.8 percent decline in births for 2020 compared with 2019, but the rate of decline was much faster at the end of the year (8 percent in December), suggesting dramatic early effects of the COVID-19 pandemic, which began affecting social life in late March 2020. Using birth data from Florida and Ohio counties through February 2021, this analysis examines whether and how much falling birth rates were associated with local pandemic conditions, specifically infection rates and reductions in geographic mobility. Results show that the vast majority of counties experienced declining births, suggestive of a general influence of the pandemic, but also that declines were steeper in places with greater prevalence of COVID-19 infections and more extensive reductions in mobility. The latter result is consistent with more direct influences of the pandemic on family planning or sexual behavior. The idea that social isolation would cause an increase in subsequent births receives no support.

Cytokines and chemokines are important regulators of airway hyper-responsiveness, immune cell infiltration, and inflammation and are produced when mast cells are stimulated with interleukin-33 (IL-33). Here, we establish that the salt-inducible kinases (SIKs) are required for the IL-33-stimulated transcription of il13, gm-csf and tnf and hence the production of these cytokines. The IL-33-stimulated secretion of IL-13, granulocyte-macrophage colony stimulating factor, and tumor necrosis factor was strongly reduced in fetal liver-derived mast cells from mice expressing a kinase-inactive mutant of SIK3 and abolished in cells expressing kinase-inactive mutants of SIK2 and SIK3. The IL-33-dependent secretion of these cytokines and several chemokines was also abolished in SIK2/3 double knock-out bone marrow-derived mast cells (BMMC), reduced in SIK3 KO cells but little affected in BMMC expressing kinase-inactive mutants of SIK1 and SIK2 or lacking SIK2 expression. In SIK2 knock-out BMMC, the expression of SIK3 was greatly increased. Our studies identify essential roles for SIK2 and SIK3 in producing inflammatory mediators that trigger airway inflammation. The effects of SIKs were independent of IκB kinase β, IκB kinase β-mediated NF-κB-dependent gene transcription, and activation of the mitogen-activated protein kinase family members p38α and c-jun N-terminal kinases. Our results suggest that dual inhibitors of SIK2 and SIK3 may have therapeutic potential for the treatment of mast cell-driven diseases.

In this research article, we draw upon major reports from a cross-section of disciplines related to large scale scientific information ecosystems to characterize the most significant research challenges, and promising potential approaches. We explore two themes that emerge across research areas: the need to align research approaches and methods with core ethical principles; and the promise of approaches that are transdisciplinary and cross-sectoral.

Components of bacteria and viruses activate Toll-Like Receptors in host cells, triggering the formation of the Myddosome and a signalling network that culminates in the production and release of the inflammatory mediators required to combat pathogenic infection. The Myddosome initiates signalling by recruiting and activating five E3 ligases that generate hybrid ubiquitin chains and attach them to components of the Myddosome. These ubiquitin chains act as a scaffold for the recruitment and activation of ubiquitin-binding proteins, which include the "master" protein kinases TAK1 and IKKβ that drive inflammatory mediator production, as well as other proteins like ABIN1 and A20 that restrict activation of the network to prevent the overproduction of these substances that can lead to autoimmunity and organ damage. Here we review recent developments in our understanding of this network, focusing on the unexpected discovery that the E3 ligase HOIL-1 initiates the formation of hybrid ubiquitin chains by forming an ester bond between the first ubiquitin and the protein components of the Myddosome.

Resume La Haute Autorite de sante a edite des regles concernant la prise en charge medicale des patients transsexuels. Ces patients doivent etre evalues par un psychiatre et un endocrinologue avant la chirurgie. Un delai d’au moins un an doit etre respecte entre la demande et l’intervention selon le Conseil national de l’Ordre des medecins. Les patients operes en milieu hospitalier public peuvent etre pris en charge par le systeme de l’assurance maladie sous couvert du respect de la procedure de prise en charge par un plasticien, un endocrinologue et un psychiatre durant au moins deux ans. Les medecins peuvent faire jouer la clause de conscience si le patient peut etre refere a un autre medecin de son choix. Il est recommande que le patient soit adresse en consultation d’anesthesie avec une lettre detaillee de son parcours medical anterieur.

Nobel-winning biochemist who discovered a ubiquitous cell-regulatory mechanism. Nobel-winning biochemist who discovered a ubiquitous cell-regulatory mechanism.

The adaptor protein MyD88 is required for signal transmission by toll-like receptors and receptors of the interleukin-1 family of cytokines. MyD88 signalling triggers the formation of Lys63-linked and Met1-linked ubiquitin (K63-Ub, M1-Ub) chains within minutes. The K63-Ub chains, which are formed by the E3 ubiquitin ligases TRAF6, Pellino1 and Pellino2, activate TAK1, the master kinase that switches on mitogen-activated protein (MAP) kinase cascades and initiates activation of the canonical IκB kinase (IKK) complex. The M1-Ub chains, which are formed by the linear ubiquitin chain assembly complex (LUBAC), bind to the NEMO (NF-κB essential modulator) component of the IKK complex and are required for TAK1 to activate IKKs, but not MAP kinases. An essential E3 ligase-independent role of TRAF6 is to recruit LUBAC into the MyD88 signalling complex, where it recognises preformed K63-Ub chains attached to protein components of these complexes, such as IRAK1 (IL-1 receptor-associated kinase), producing ubiquitin chains containing both types of linkage, termed K63/M1-Ub hybrids. The formation of K63/M1-Ub hybrids, which is a feature of several innate immune signalling pathways, permits the co-recruitment of proteins that interact with either K63-Ub or M1-Ub chains. Two likely roles for K63/M1-Ub hybrids are to facilitate the TAK1-dependent activation of the IKK complex and to prevent the hyperactivation of these kinases by recruiting A20 and A20-binding inhibitor of NF-κB1 (ABIN1). These proteins restrict activation of the TAK1 and IKK complexes, probably by competing with them for binding to K63/M1-Ub hybrids. The formation of K63/M1-Ub hybrids may also regulate the rate at which the ubiquitin linkages in these chains are hydrolysed. The IKK-catalysed phosphorylation of some of its substrates permits their recognition by the E3 ligase SCFβTRCP, leading to their Lys48-linked ubiquitylation and proteasomal degradation. Innate immune signalling is therefore controlled by the formation and destruction of three different types of ubiquitin linkage.