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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.

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