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Societal Impact Statement Debates about the impacts of human settlement on Madagascar's habitat have missed the Malagasy perspective. Using indigenous and local knowledge in the form of toponyms, we find many regions across today's treeless grasslands are named after forest/trees, suggesting they may be novel. Where observed habitat does not match toponyms, erosion is significantly more likely at landscape scales. This suggests rapid expansion of erosion following human removal and burning of endemic forest, savanna woodland, heathland and grass assemblages. Findings also provide a timely warning: current practices may be unsustainable and may impact not only biodiversity but also human wellbeing unless urgently addressed. Summary The debate surrounding the extent of Madagascar's treeless grasslands at human settlement is important because introduced disturbance can negatively impact the biodiversity and productivity of systems that evolved under different regimes. Indigenous and local knowledge (ILK), often overlooked, can provide information about past vegetation structure. To test whether clearance of forest and trees, frequent fires and pasture preparation have accelerated today's island‐wide erosion, we use vegetation toponyms and assess whether mismatches between these and current vegetation types are significantly more likely to be associated with erosion. Using Malagasy and Imerina linguistic records spanning 150 years, we mapped forest‐related and grassland‐related extensive toponyms in current grassland and forest, respectively. We then assessed whether remotely‐sensed erosion was more likely when toponyms and current habitat did not match. We found 316 sites in treeless grasslands, named after forest/clusters of trees, but no grassland‐named sites in forest. Globally, natural forest and grasslands both constrain erosion. Forest toponyms in grassland were significantly more likely to reflect erosion than sites in extant forest. These findings concur with palaeoecological, geomorphological, molecular and rangeland palatability studies. Malagasy ILK, hitherto largely ignored as a source of information, strongly suggests vegetation clearance and human disturbance have exacerbated the degradation of terrestrial, freshwater and marine ecosystems through topsoil loss and siltation and selected fire‐adapted, less‐palatable grasslands. Malagasy ILK in the form of toponyms highlights the need to address the negative impacts of burning and land‐clearance practices (e.g., loss of biodiversity and ecological function, decreased agricultural productivity, collapsed fisheries), given island‐wide challenges to food security and conservation.

Abstract A significant effort in biomedical sciences has been made to examine relationships between sex and the mechanisms underlying various disease states and behaviors, including sleep. Here, we investigated biological sex differences in sleep using male and female C57BL/6J mice (n = 267). Physiological parameters were recorded for 48-h using non-invasive piezoelectric cages to determine total sleep, non-rapid eye movement (NREM) sleep, rapid eye movement (REM)-like sleep, and wakefulness (WAKE). We fit hierarchical generalized linear mixed models with nonlinear time effects and found substantial sex differences in sleep. Female mice slept less overall, with less NREM sleep compared to males. Females also exhibited more REM-like sleep and WAKE and had shorter NREM sleep bout lengths. We also conducted a simulation exercise where we simulated a hypothetical treatment that altered the sleep of female mice, but not male mice. In models that included an appropriate sex by treatment interaction, a female-specific treatment response was accurately estimated when sample sizes were equal but was not detected when sample sizes were unequal, and females were underrepresented. Failure to include both sexes in experimental designs or appropriately account for sex during analysis could lead to inaccurate translational recommendations in pre-clinical sleep studies.

Abstract Background Juvenile idiopathic arthritis (JIA) comprises a heterogeneous group of conditions that can cause marked disability and diminished quality of life. Data on predictors of clinical response are insufficient to guide selection of the appropriate biologic agent for individual patients. This study aimed to investigate the propensity of S100A8/9 and S100A12 as predictive biomarkers of abatacept response in polyarticular-course juvenile idiopathic arthritis (pJIA). Methods Data from a phase 3 trial (NCT01844518) of subcutaneous abatacept in patients with active pJIA (n = 219) were used in this exploratory analysis. Association between biomarker levels at baseline and improvements in JIA-American College of Rheumatology (ACR) criteria responses or baseline disease activity (measured by Juvenile Arthritis Disease Activity Score in 27 joints using C-reactive protein [JADAS27-CRP]) were assessed. Biomarker level changes from baseline to month 4 were assessed for disease outcome prediction up to 21 months. Results At baseline, 158 patients had available biomarker samples. Lower baseline S100A8/9 levels (≤ 3295 ng/mL) were associated with greater odds of achieving JIA-ACR90 (odds ratio [OR]: 2.54 [95% confidence interval (CI): 1.25–5.18]), JIA-ACR100 (OR: 3.72 [95% CI: 1.48–9.37]), JIA-ACR inactive disease (ID; OR: 4.25 [95% CI: 2.03–8.92]), JADAS27-CRP ID (OR: 2.34 [95% CI: 1.02–5.39]) at month 4, and JIA-ACR ID (OR: 3.01 [95% CI: 1.57–5.78]) at month 16. Lower baseline S100A12 levels (≤ 176 ng/mL) were associated with greater odds of achieving JIA-ACR90 (OR: 2.52 [95% CI: 1.23–5.13]), JIA-ACR100 (OR: 3.68 [95% CI: 1.46–9.28]), JIA-ACR ID (OR: 3.66 [95% CI: 1.76–7.61]), JIA-ACR90 (OR: 2.03 [95% CI: 1.07–3.87]), JIA-ACR100 (OR: 2.14 [95% CI: 1.10–4.17]), and JIA-ACR ID (OR: 4.22 [95% CI: 2.15–8.29]) at month 16. From baseline to month 4, decreases in S100A8/9 and S100A12 generally exceeded 50% among JIA-ACR90/100/ID responders. Conclusion Lower baseline levels of S100A8/9 and S100A12 proteins predicted better response to abatacept treatment than higher levels and may serve as early predictive biomarkers in pJIA. Decreases in these biomarker levels may also predict longer-term response to abatacept in pJIA.

Abstract Greater exposure to stressors over the life course is believed to promote striatum-dependent over hippocampus-dependent learning and memory processes under stressful conditions. However, little research in this context has actually assessed lifetime stressor exposure and, moreover, it remains unknown whether greater cumulative lifetime stressor exposure exerts comparable effects on striatum-dependent learning and hippocampus-dependent learning in non-stressful contexts. To investigate this issue, we used the Stress and Adversity Inventory for Adults (Adult STRAIN) and Multicued Search Task to investigate the relation between cumulative lifetime stressor exposure and striatum-dependent stimulus–response learning and hippocampus-dependent contextual learning under non-stressful conditions among healthcare professionals (N = 205; 157 females, 48 males; Age: M = 34.23, SD 9.3, range 20–59 years). Individuals with moderate, but not low, cumulative lifetime stressor exposure exhibited impaired learning for stimulus–response associations. In contrast, learning for context associations was unrelated to participants' lifetime stressor exposure profiles. These results thus provide first evidence that cumulative lifetime stressor exposure may have negative consequences on human striatum-dependent stimulus–response learning under non-stressful environmental conditions.

This paper examines the interplay of environmental change and human demographic shifts among late Pleistocene and Holocene populations in the hyper-arid context of the Central Namib Gravel plains, Western Namibia. This paper applies a structural equation modeling (SEM) approach to arid-zone hunter-gatherer mobility patterns and settlement systems based on variability among ethnographically and historically observed groups. It then applies this model in examining issues related to the Middle-to-Later Stone Age transition at the sites of Erb Tanks and the Mirabib rock shelter. This model suggests that, among modern arid-zone foragers, population density is by far the strongest factor influencing mobility and settlement systems, with higher population densities favoring longer residential site usages and shorter residential moves within smaller territories. In contrast, environmental variables having to do with annual rainfall, rainfall seasonality, and effective temperature affect mobility and settlement systems in ambiguous and statistically insignificant ways. This paper closes by arguing that regional-scale population increases signaled by various features of Later Stone Age archaeological patterning led to local population expansions in the Central Namib Desert, and that this in turn caused a range of recognizable cultural shifts that were strongly linked with adaptations to aridity.

Societal Impact Statement To better understand the impact of human settlement on vegetation distribution and disturbance regimes in Madagascar, the 2021 Malagasy Grassy Biomes Workshop called for an examination of the role of extinct megafauna in determining the historic distribution of habitat mosaics across regions that are today dominated by vast treeless grasslands. A systematic literature review facilitated the reconstruction and evaluation of giant tortoise habitat, feeding habitats, diets and Aldabrachelys tortoise impacts on disturbance regimes. In conclusion, reintroduction of mixed‐feeding Aldabrachelys (the diets of which were often dominated by C3 woody material, but never by C4 grasses, across its range), will likely impact plant species composition, select a tree‐rich habitat mosaic, reduce fire frequency, re‐establish lost ecological functions and improve landscape productivity at a range of scales, including rangeland palatability, benefitting both biodiversity and human wellbeing. Summary Grasslands with little tree cover today comprise 80% of Madagascar's habitat. Determining their extent at human settlement can guide ecological restoration and enhance human well‐being, so the 2021 Malagasy Grassy Biomes Workshop identified the role of extinct megafauna in determining habitat as a critical knowledge gap. Using a systematic literature review, combined with extracted datasets, we address this, examining anticipated habitat selection by giant tortoises following reintroduction to Madagascar (where the Aldabran giant tortoise, Aldabrachelys gigantea, provides ecological functions lost when A. abrupta and A. grandidieri went extinct). When comparing current and historical tortoise selection of habitat across the Mascarenes and Aldabra with contemporary Malagasy habitat, areas in Madagascar where giant tortoises historically ranged, today have a significantly different habitat composition to the forested habitat that supported giant tortoises on other islands. Dietary 13C isotope ratios show that Malagasy Aldabrachelys and Mascarene tortoises were mixed feeders, with diets often dominated by C3 woody intake, but never by C4 grasses. Across systems, giant tortoises required and selected, tree‐rich habitat mosaics, different to current pastoralist fire‐selected Malagasy grasslands characterized by sparse tree cover. Furthermore, Aldabran Aldabrachelys tortoise turf, restricted to small areas (large tracts of unshaded vegetation present physiological challenges to Aldabrachelys' survival), is compositionally different to Malagasy and African obligate C4 grazing lawns. Ecological, palaeoecological, geomorphological and molecular evidence support a lost Malagasy habitat mosaic where hippo and tortoise diets were C3‐dominated, because they inhabited closed‐canopy systems, with abutting open‐canopy areas harbouring endemic‐rich, C4 grassy understories and limited grasslands. The review suggests that rewilding with A. gigantea will help restore ecological functions, productivity and landscape‐scale degradation lost through cattle‐based pastoralism, re‐establish tree‐rich habitat mosaics, and mitigate against frequent bushfires, benefiting biodiversity and humans at multiple scales.

Societal Impact Statement The relationship between rainfall, fire and habitat can display incongruencies. The 2021 Malagasy Grassy Biomes Workshop identified understanding fire regimes as a knowledge gap. This study pinpoints regions where anthropogenic fire has the potential to transform or has transformed habitat to treeless‐grassland, by identifying landscape‐scale, island‐wide fire anomalies across half of Madagascar. Its eastern forests burn like savannas, and central‐western grasslands burn frequently and intensely despite receiving rainfall usually associated with forest and fire‐absence. Recognising the incongruity and better understanding its drivers can mitigate against landscape‐scale degradation, improving ecological function, and human well‐being. Summary Data show that since 1953, human‐lit fires on Madagascar have transformed clear‐cut forest to treeless‐grasslands. To address the extent of Malagasy treeless‐grasslands at human settlement, the 2021 Malagasy Grassy Biomes Workshop identified the role of fire as a critical knowledge‐gap for understanding ecological function. The relationship between mean annual precipitation (MAP), fire and habitat is well established across mesic systems. Anthropogenically transformed habitats often deviate from expected ecological patterns, so we tested for landscape‐scale, island‐wide MAP‐related fire and habitat anomalies. We collated Malagasy fire, habitat and MAP datasets, identifying location and scale of incongruities relative to global fire‐habitat‐MAP expectations. Next, we tested for mismatches in fire regimes (frequency, timing, extent and intensity of fires) between Malagasy and equivalent global biomes, using global, comprehensive landscape‐scale fire regime data. Across half of Madagascar, fire frequency and habitat are decoupled from MAP, and fire regimes across Malagasy ecoregions differ significantly from those in shared biomes elsewhere in the world. Landscape‐scale incongruities span Malagasy eastern forests (which burn like savanna systems) and central‐western treeless‐grasslands, which burn frequently and intensely despite receiving MAP typical of forest presence and fire‐absence, globally. Fire‐MAP incongruities identify potentially transformed areas, or those undergoing transformation by fire, and establish a platform for investigating the nuanced social, political and ecological dynamics that may contribute to and perpetuate these anomalies. Incongruities also highlight the anthropogenic landscape degradation associated with fire anomalies. Addressing these impacts can facilitate restoration of ecological function, productivity and food security, benefiting biodiversity and humans at multiple scales.

AbstractA growing body of work has found that a mismatch between early and recent life stress, more than a cumulative influence of stress, contributes to detrimental stress-related health outcomes. To date, however, no work has examined how such a mismatch might relate to stress-related cognitive outcomes. We addressed this gap in the current study by assessing participants’ (N = 154, Mage = 18.7, 104 female) early and recent life stress using the same inventory, and subsequently assessing their inhibitory control in a hybrid stop-signal/flanker task. Surprisingly, we found that a greater degree of stressor mismatch was associated with better response inhibition (i.e. smaller stop-signal reaction time) across a number of analytic approaches. Cognitive inhibition (i.e. the flanker interference effect) was not associated with stressor mismatch. These results thus show that a greater degree of mismatch between early and recent life stress is related to response inhibition in the same way as acute stress affects response inhibition, suggesting that response inhibition may be an important cognitive process for navigating both acute stress and general environmental conditions that do not match the conditions in which expected stress occurrence was established.

IntroductionInsufficient or disturbed sleep is strongly associated with adverse health conditions, including various neurodegenerative disorders. While the relationship between sleep and neurodegenerative disease is likely bidirectional, sleep disturbances often predate the onset of other hallmark clinical symptoms. Neuronal waste clearance is significantly more efficient during sleep; thus, disturbed sleep may lead to the accumulation of neuronal proteins that underlie neurodegenerative diseases. Key pathological features of neurodegenerative diseases include an accumulation of misfolded or misprocessed variants of amyloid beta (Aβ), tau, alpha synuclein (α-syn), and TarDNA binding protein 43 (TDP-43). While the presence of fibrillar protein aggregates of these neuronal proteins are characteristic of neurodegenerative diseases, the presence of small soluble toxic oligomeric variants of these different proteins likely precedes the formation of the hallmark aggregates.MethodsWe hypothesized that sleep deprivation would lead to accumulation of toxic oligomeric variants of Aβ, tau, α-syn, and TDP-43 in brain tissue of wild-type mice. Adult mice were subjected to 6 h of sleep deprivation (zeitgeber 0–6) for 5 consecutive days or were left undisturbed as controls. Following sleep deprivation, brains were collected, and protein pathology was assessed in multiple brain regions using an immunostain panel of reagents selectively targeting neurodegenerative disease-related variants of Aβ, tau, α-syn, and TDP-43.ResultsOverall, sleep deprivation elevated levels of all protein variants in at least one of the brain regions of interest. The reagent PDTDP, targeting a TDP-43 variant present in Parkinson’s disease, was elevated throughout the brain. The cortex, caudoputamen, and corpus callosum brain regions showed the highest accumulation of pathology following sleep deprivation.DiscussionThese data provide a direct mechanistic link between sleep deprivation, and the hallmark protein pathologies of neurodegenerative diseases, such as Alzheimer’s and Parkinson’s diseases.

Diffuse large B-cell lymphoma (DLBCL) is the most common malignancy that develops in patients with ataxia-telangiectasia, a cancer-predisposing inherited syndrome characterized by inactivating germline ATM mutations. ATM is also frequently mutated in sporadic DLBCL. To investigate lymphomagenic mechanisms and lymphoma-specific dependencies underlying defective ATM, we applied ribonucleic acid (RNA)-seq and genome-scale loss-offunction clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 screens to systematically interrogate B-cell lymphomas arising in a novel murine model (Atm-/-nu-/-) with constitutional Atm loss, thymic aplasia but residual T-cell populations. Atm-/-nu-/-lymphomas, which phenotypically resemble either activated B-cell-like or germinal center Bcell-like DLBCL, harbor a complex karyotype, and are characterized by MYC pathway activation. In Atm-/-nu-/-lymphomas, we discovered nucleotide biosynthesis as a MYCdependent cellular vulnerability that can be targeted through the synergistic nucleotidedepleting actions of mycophenolate mofetil (MMF) and the WEE1 inhibitor, adavosertib (AZD1775). The latter is mediated through a synthetically lethal interaction between RRM2 suppression and MYC dysregulation that results in replication stress overload in Atm-/-nu-/-lymphoma cells. Validation in cell line models of human DLBCL confirmed the broad applicability of nucleotide depletion as a therapeutic strategy for MYC-driven DLBCL independent of ATM mutation status. Our findings extend current understanding of lymphomagenic mechanisms underpinning ATM loss and highlight nucleotide metabolism as a targetable therapeutic vulnerability in MYC-driven DLBCL.

Societal Impact Statement The 26th United Nations Climate Change Conference emphasised the need to modify practices that negatively impact biodiversity and food security in the context of global change. Following Madagascar's drought‐induced famine, our systematic review supports the theory that grasslands of the Malagasy Central Highland that are subjected to human‐lit fires are anthropogenically derived. Furthermore, these overly frequent fires that characterise much of the Malagasy Central Highland grasslands select poorly palatable grasses. Given the reliance on pastoralism as insurance against crop failure in Madagascar, fire‐dependent practices that degrade rangeland emerge as a threat to food security and biodiversity. Education can mitigate against future humanitarian crises. Summary Food insecurity is greatest in countries where impacts of global change are predicted to be severe. Many, like Madagascar, rely on livestock‐based pastoralism (and consequently palatable rangelands) for insurance against natural disasters and crop failure. It is recognised that derived grasslands can impact climate and biodiversity. Furthermore, the well‐established palatability‐flammability trade‐off predicts that overly‐frequent fires select increasingly unpalatable, fire‐adapted grassland. The drought‐induced Malagasy famine of 2021 highlights the need to identify factors that threaten food security. Given the ubiquitous practice of rangeland preparation through annual, landscape‐scale human‐lit fires, we evaluate whether Malagasy grasslands are derived and then test for fire‐driven selection of increasingly degraded and unpalatable rangelands across Madagascar's largest grassland system, the Malagasy Central Highland (MCH). We conducted a systematic literature review, evaluating for a palatability‐flammability trade‐off, by determining dominant Malagasy grass species, and then applying functional traits, and palatability ratings to these species. Data were extracted using a suite of relevant search terms, and of 1977 studies identified, 145 were directly relevant to the questions posed. Evidence from the review is compelling for much of the Malagasy highland grassland being derived. Furthermore, Malagasy dominant grass species are fire‐adapted with poor forage‐value, suggesting current burning practices negatively impact both biodiversity and pastoralism. Decreasing rangeland palatability caused by human‐lit fires in a society suffering food insecurity emphasises the need to re‐evaluate pastoralist burning practices. Identifying optimal fire frequencies can avert breaching fire‐induced tipping points to rangeland palatability and the humanitarian crises that may follow.

Abstract Multi-system inflammatory syndrome in children (MIS-C) is a severe post-acute sequela of SARS-CoV-2 infection in children, and there is a critical need to unfold its highly heterogeneous disease patterns. Our objective was to characterize the illness spectrum of MIS-C for improved recognition and management. We conducted a retrospective cohort study using data from March 1, 2020–September 30, 2022, in 8 pediatric medical centers from PEDSnet. We included 1139 children hospitalized with MIS-C and used their demographics, symptoms, conditions, laboratory values, and medications for analyses. We applied heterogeneity-adaptive latent class analyses and identified three latent classes. We further characterized the sociodemographic and clinical characteristics of the latent classes and evaluated their temporal patterns. Class 1 (47.9%) represented children with the most severe presentation, with more admission to the ICU, higher inflammatory markers, hypotension/shock/dehydration, cardiac involvement, acute kidney injury and respiratory involvement. Class 2 (23.3%) represented a moderate presentation, with 4–6 organ systems involved, and some overlapping features with acute COVID-19. Class 3 (28.8%) represented a mild presentation. Our results indicated that MIS-C has a spectrum of clinical severity ranging from mild to severe and the proportion of severe or critical MIS-C decreased over time.

Acute stress has been well-established to impair working memory. However, less is known about how writing about an unresolved stressor may influence working memory or working memory processes. We addressed these issues in the present study (N = 282) by randomly assigning participants to write about an unresolved stressful experience (stressful writing condition or the events of the previous day). We then both measured performance on a change detection task and used computational modeling to estimate the processes underlying performance: attention, capacity, and guessing bias. We found that, relative to the control condition, writing about a stressful experience impaired change detection task performance and significantly impaired task attention. These results show that the effects of writing about an unresolved stressor may mimic the effects of acute stress on working memory, rather than conforming to expectations from mood-as-information theory.

Our density functional theory calculations (by GAUSSIAN) on the Si4O104− moiety and Na4Si4O10 molecule reproduce the experimental Raman frequencies and relative intensities for the three A1 modes in the experimental Raman spectra of Na2Si2O5 and BaSi2O5 crystals and glasses. Taking the center of mass into consideration, accompanied by visualization using GaussView, these calculations provide a firm assignment of these three peaks: the 1060 cm−1 peak to the Si-NBO stretch (NBO = Si–O or Si–O–Na), the ∼600 cm−1 peak to the Si-BO stretch (BO = Si–O–Si), and the ∼500 cm−1 peak to the Si-BO-Si bend. Our GAUSSIAN calculations on modified “non-equilibrium” SiO44− tetrahedra, constrained to have the same Si–O bond lengths and O–Si–O bond angles as in the Si4O104− moiety, confirm the above-mentioned assignment and show reasonable agreement with the experimental Raman frequencies and relative intensities. We then examine the regular trends in the Si-NBO and Si-BO stretch frequencies for all Qn (n = 0–4) species in silicate glasses and crystals. The large systematic change in the differences between these frequencies (Δ1) is explained by electronic changes within the SiO4 tetrahedron. For example, the larger Δ1 for the Q3 tetrahedra than the Q2 tetrahedra is due to the changes in Si, BO, and NBO electron densities on the tetrahedral Si and O atoms, as measured by Si 2p and O 1s x-ray photoelectron spectra. This trend strongly suggests that the Si-BO stretch frequency for the Q4 species in v-SiO2 should be ∼500 cm−1, close to the A1 breathing mode frequency of 495 cm−1 for four membered rings previously assigned.

Our new density functional theory calculations by Gaussian reproduce the bond lengths and Raman spectra of a number of model tetrahedral Si and P compounds [the ten compounds in the SiHxD4−x and SiFxCl4−x (x = 0–4) series, and the tetramer P4O10]. The number of symmetric A1 peaks is determined by group theory, and the center of mass is especially important for the assignment of the three A1 peaks for P4O10 to the two P–O stretches and the O–P–O bend. Additional Gaussian calculations on isolated C3v PO4 tetrahedra, using the P–O bond lengths and O–P–O bond angles from the P4O10 optimization, also reproduce the three peak P4O10 Raman spectra and provide an important guide to the assignment of the two low energy peaks at ∼500 and ∼700 cm−1 to the P–BO (BO = P–O–P) stretch and BO–P–BO bend, respectively, in contrast to all earlier assignments. In the SiFxCl4−x series, there is a regular increase in the Si–F A1 frequency and a decrease in the Si–Cl A1 frequency across the series. Similar trends are seen in the SiClx(CH3)4−x series, and these trends are due to changes in the electron densities in the tetrahedron when an electronegative F or Cl is replaced by a less electronegative Cl or CH3, as measured by previous Si 2p, F 1s, Cl 2p, and C 1s x-ray photoelectron spectroscopy. These considerations serve as a guide for the interpretation of the Raman spectra of silicate glasses in Paper II [Bancroft et al., AIP Adv. 13, 125216 (2023)].

Abstract Macrophage activation syndrome (MAS) is a life-threatening complication of pediatric rheumatic diseases, occurring most commonly in children with systemic juvenile idiopathic arthritis (SJIA). Despite several classes of currently available treatment options for SJIA, including biologic agents targeting IL-1 or IL-6, there remain severe cases suffering from refractory disease and recurrent MAS. The phenotype of MAS is similar to hemophagocytic lymphohistiocytosis (HLH), but the underlying pathophysiology of MAS complicating SJIA or other disorders has not been fully clarified. These facts make it challenging to develop and utilize animal models to study MAS. To date, there is no “perfect” model replicating MAS, but several models do demonstrate aspects of SJIA and/or MAS. In this review, we examine the proposed animal models of SJIA and MAS, focusing on how they reflect these disorders, what we have learned from the models, and potential future research questions. As we better understand the key features of each, animal models can be powerful tools to further define the pathophysiology of SJIA and MAS, and develop new treatment targets and strategies.

Systemic autoimmune and autoinflammatory diseases are characterized by genetic and cellular heterogeneity. While current single-cell genomics methods provide insights into known disease subtypes, these analysis methods do not readily reveal novel cell-type perturbation programs shared among distinct patient subsets. Here, we performed single-cell RNA-Seq of PBMCs of patients with systemic juvenile idiopathic arthritis (SJIA) with diverse clinical manifestations, including macrophage activation syndrome (MAS) and lung disease (LD). We introduced two new computational frameworks called UDON and SATAY-UDON, which define patient subtypes based on their underlying disrupted cellular programs as well as associated biomarkers or clinical features. Among twelve independently identified subtypes, this analysis uncovered what we believe to be a novel complement and interferon activation program identified in SJIA-LD monocytes. Extending these analyses to adult and pediatric lupus patients found new but also shared disease programs with SJIA, including interferon and complement activation. Finally, supervised comparison of these programs in a compiled single-cell pan-immune atlas of over 1,000 healthy donors found a handful of normal healthy donors with evidence of early inflammatory activation in subsets of monocytes and platelets, nominating possible biomarkers for early disease detection. Thus, integrative pan-immune single-cell analysis resolved what we believe to be new conserved gene programs underlying inflammatory disease pathogenesis and associated complications.

A comprehensive two-dimensional gas chromatography instrument with quadrupole mass spectrometry (GC×GC-qMS) is developed and evaluated, by retrofitting a commercial GC-qMS instrument with a high temperature diaphragm valve (HT-DV) modulator. A porous layer open tubular (PLOT) column in the first dimension (1D), and a more polar wall coated open tubular (WCOT) column in the second dimension (2D) were used. The HT-DV can modulate analytes of any volatility and the combination of the PLOT and WCOT columns provides ample separation selectivity for samples containing an extended range of volatility from highly volatile analytes to analytes of more moderate volatility. In addition to instrument development, we describe data visualization considerations when upgrading from GC-qMS to GC×GC-qMS. Unlike time-of-flight mass spectrometry (TOFMS), the mass spectrum cycle time for qMS (defined as the sum of scan time and interscan delay) is a linear function of the range of mass channels (m/z) analyzed. If the cycle time does not divide evenly into the modulation period, PM, visual distortions will appear in the GC×GC chromatogram. We present two approaches to address this GC×GC chromatogram distortion issue. The first method does not require post-run computations; however, care must be taken in selecting an appropriate mass range to minimize the visual distortion issue. A half of the modulation period of total shift in the 2D separations with a PM of 2 s was determined tolerable for a 15 min separation, with 44 mass ranges (and their accompanying scan rate) deemed suitable out of 350 possible mass ranges. Also, a computational “resampling” method, which can be used to correct any qMS scan rate used was implemented. Figures-of-merit for the HT-DV modulated GC×GC-qMS instrument were also examined. Reproducible peak height (average RSDheight = 9.2%, s = 3.5%) and areas (average RSDarea = 9.3%, s = 2.8%) were obtained with low LODs (average LOD = 4.2 mg/L, s = 2.5 mg/L).

The SMC5/6 complex is critical for genome stability. Here, the authors identify mutations in SLF2 and SMC5 as cause of Atelís Syndrome characterized by microcephaly, short stature, anemia, segmented chromosomes and mosaic variegated hyperploidy.

Attentional biases toward or away from emotionally evocative stimuli have been well documented and are known to be clinically relevant, making it important to understand how various factors contribute to them. Some work has suggested that acute stress modulates attentional biases, but this work has produced inconsistent results. For example, many studies have found that stress enhances attentional bias, others that stress decreases attentional bias, and others still that there is no effect of stress at all. Methodological differences may explain these inconsistencies. For example, discrepancies exist between studies in participant sex (e.g., mixed sample vs. all men) and in the type of attentional bias paradigm. We addressed these gaps by examining the effects of an acute social stressor (vs. control) on attentional bias assessed via facial dot probe, focusing on potential sex differences in these effects (N = 141). We found that, overall, participants were significantly biased towards threat, but biases did not differ by stress condition or sex. These findings help to clarify the existing discrepancy in the literature, as we found that stress exerts little if any effect on attentional bias assessed via a facial dot probe.

Axons crossing the CNS midline regulate their responsiveness to both attractive and repulsive cues. In this issue, Dailey-Krempel et al. find different modes of action for DCC isoforms and uncover evidence against the silencing model of axon guidance.

Although protein hydroxylation is a relatively poorly characterized posttranslational modification, it has received significant recent attention following seminal work uncovering its role in oxygen sensing and hypoxia biology. Although the fundamental importance of protein hydroxylases in biology is becoming clear, the biochemical targets and cellular functions often remain enigmatic. JMJD5 is a “JmjC-only” protein hydroxylase that is essential for murine embryonic development and viability. However, no germline variants in JmjC-only hydroxylases, including JMJD5, have yet been described that are associated with any human pathology. Here we demonstrate that biallelic germline JMJD5 pathogenic variants are deleterious to JMJD5 mRNA splicing, protein stability, and hydroxylase activity, resulting in a human developmental disorder characterized by severe failure to thrive, intellectual disability, and facial dysmorphism. We show that the underlying cellular phenotype is associated with increased DNA replication stress and that this is critically dependent on the protein hydroxylase activity of JMJD5. This work contributes to our growing understanding of the role and importance of protein hydroxylases in human development and disease.

ABSTRACT Variants of concern (VOC) in SARS-CoV-2 refer to viruses whose viral genomes differ from the ancestor virus by ≥3 single-nucleotide variants (SNVs) and that show the potential for higher transmissibility and/or worse clinical progression. VOC have the potential to disrupt ongoing public health measures and vaccine efforts. Still, too little is known regarding how frequently new viral variants emerge and under what circumstances. We report a study to determine the degree of SARS-CoV-2 sequence evolution in 94 patients and to estimate the frequency at which highly diverse variants emerge. Two cases accumulated ≥9 SNVs over a 2-week period and one case accumulated 23 SNVs over 3 weeks, including three nonsynonymous mutations in the spike protein (D138H, E554D, D614G). The remainder of the infected patients did not show signs of intra-host evolution. We estimate that in as much as 2% of hospitalized COVID-19 cases, variants with multiple mutations in the spike glycoprotein emerge in as little as 1 month of persistent intra-host virus replication. This suggests the continued local emergence of variants with multiple nonsynonymous SNVs, even in patients without overt immune deficiency. Surveillance by sequencing for (i) viremic COVID-19 patients, (ii) patients suspected of reinfection, and (iii) patients with diminished immune function may offer broad public health benefits. IMPORTANCE New SARS-CoV-2 variants can potentially disrupt ongoing public health measures and vaccine efforts. Still, little is known regarding how frequently new viral variants emerge and under what circumstances. Based on this study, we estimate that in hospitalized COVID-19 cases, variants with multiple mutations may emerge locally in as little as 1 month, even in patients without overt immune deficiency. Surveillance by sequencing for continuously shedding patients, patients suspected of reinfection, and patients with diminished immune function may offer broad public health benefits.

Macrophage activation syndrome (MAS) is a subtype of haemophagocytic lymphohistiocytosis (HLH), and a well-described complication of systemic juvenile idiopathic arthritis (SJIA), triggered by disease onset or flare, infection, or some medications. Here, we report a 20-year-old man with previously well-controlled SJIA, who developed first time MAS after acute Epstein-Barr virus (EBV) infection, with MAS recurrence due to a drug reaction, ‘3-week sulfasalazine syndrome’, secondary to prophylactic trimethoprim/sulfamethoxazole. Both episodes of MAS were minimally responsive to pulse corticosteroids. Initial EBV-driven MAS was treated with multiple doses of emapalumab prior to resolution, while MAS secondary to sulfasalazine-induced 3-week syndrome required the initiation of ruxolitinib. This case exhibits two rare but life-threatening causes of MAS/secondary HLH in a single patient and the difficulties in their diagnosis and management.

Abstract Uncertainty is inherent in most decisions humans make. Economists distinguish between two types of decision-making under non-certain conditions: those involving risk (i.e., known outcome probabilities) and those that involve ambiguity (i.e., unknown outcome probabilities). Prior research has identified individual differences that explain risk preferences, but little is known about factors associated with ambiguity aversion. Here, we hypothesized that cumulative exposure to major psychosocial stressors over the lifespan might be one factor that predicts individuals’ ambiguity aversion. Across two studies (Study 1: n = 58, M age = 25.7; Study 2: n = 188, M age = 39.81), we used a comprehensive lifetime stressor exposure inventory (i.e., the Stress and Adversity Inventory for Adults, or STRAIN) and a standard economic approach to quantify risk and ambiguity preferences. Greater lifetime stressor exposure as measured by the STRAIN, particularly in early life, was associated with higher aversion to ambiguity but not risk preferences.

The adenovirus C5 E1B-55K protein is crucial for viral replication and is expressed early during infection. It can interact with E4orf6 to form a complex that functions as a ubiquitin E3 ligase. This complex targets specific cellular proteins and marks them for ubiquitination and, predominantly, subsequent proteasomal degradation. E1B-55K interacts with various proteins, with p53 being the most extensively studied, although identifying binding sites has been challenging. To explain the diverse range of proteins associated with E1B-55K, we hypothesized that other binding partners might recognize the simple p53 binding motif (xWxxxPx). In silico analyses showed that many known E1B-55K binding proteins possess this amino acid sequence; therefore, we investigated whether other xWxxxPx-containing proteins also bind to E1B-55K. Our findings revealed that many cellular proteins, including ATR, CHK1, USP9, and USP34, co-immunoprecipitate with E1B-55K. During adenovirus infection, several well-characterized E1B-55K binding proteins and newly identified interactors, including CSB, CHK1, and USP9, are degraded in a cullin-dependent manner. Notably, certain binding proteins, such as ATR and USP34, remain undegraded during infection. Structural predictions indicate no conservation of structure around the proposed binding motif, suggesting that the interaction relies on the correct arrangement of tryptophan and proline residues.

Abstract Objective To determine whether septal perforations have an effect on nasal swell body (NSB) size. Study Design Retrospective cohort study. Setting Two tertiary academic medical centers. Methods Computed tomography maxillofacial scans of 126 patients with septal perforation and 140 control patients from November 2010 to December 2020 were evaluated. Perforation etiology was determined. Measurements included perforation length and height and swell body width, height, and length. Swell body volume was calculated. Results The width and volume of the NSB are significantly smaller in perforation patients when compared to controls. The swell body is significantly smaller and thinner in perforations exceeding 14 mm in height compared to small perforations. Perforation etiology groupings into prior septal surgery, septal trauma, septal inflammatory, and mucosal vasoconstriction categories all demonstrated decreased swell body volume and width compared to controls. Inflammatory etiology had the greatest decrease in swell body size. The hemi‐swell body on the contralateral side of a septal deviation is significantly thicker than the ipsilateral side. Conclusion The NSB is smaller in patients with septal perforation regardless of perforation size or etiology.

Post-translational modifications are critical for regulating the DNA damage response. Here, the authors identify a methylation-deubiquitination crosstalk between methyltransferase PRMT1 and deubiquitinase USP11, showing that the enzymes regulate each other’s functions in DNA repair.

Abstract Background Preterm birth has been linked to an elevated risk of heart failure and cardiopulmonary disease later in life. With improved neonatal care and survival, most infants born preterm are now reaching adulthood. In this study, we used 4D flow cardiovascular magnetic resonance (CMR) coupled with an exercise challenge to assess the impact of preterm birth on right heart flow dynamics in otherwise healthy adolescents and young adults who were born preterm. Methods Eleven young adults and 17 adolescents born preterm (

Abstract Woody structure, particularly that created by large trees, is crucial to savanna biodiversity pattern and process. We assessed changes in vertical and horizontal canopy over 15 years of saplings of Vachellia (Acacia) erioloba (camelthorn), a keystone species and one of the only trees to grow large in arid savanna on low rainfall areas of Kalahari sands, southern Africa. We also assessed whether the fastest growing saplings over the 2003–2004 growing season were also the fastest growing over the following 15 years. We measured height, canopy extent, and diameter of largest stem at ground level of 120 saplings of V. erioloba in November 2003, remeasuring these in March 2004. In November 2019, we were able to find 67 of these saplings, and we measured them again to assess growth between 2004 and 2019. We modeled interindividual explanatory variables for growth (i.e., measures taken in 2004 of height, diameter, number of stems, evidence of fire, and largest stem diameter). We also compared residuals of growth between 2003 and 2004 with those between 2004 and 2019, to assess whether individuals showed similar progress relative to their peers over the two periods considered. Height increase was extremely variable, but mean increase over 15 years was only 45.4 mm, similar to that measured for the 2003–2004 growing season (43.1 mm). Mean horizontal canopy showed far greater growth (mean 120.2 mm over 15 years, compared with 7.12 mm over 2003–2004) than vertical growth. Individuals that had shown the fastest increase in horizontal canopy in 2003–2004 tended to be the same individuals that grew fastest over the following 15 years, but even the highest “achievers” showed increases of only 47.2 mm and 37.5 mm year−1 in average horizontal canopy extent and height, respectively. This study demonstrates how crucial long‐term studies are, particularly in arid systems. Many individuals may be trapped in the “sapling” class and could take decades to become large trees. Woody sapling growth was not cumulative, and modeling studies in savannas should be cautious in extrapolating growth from one season to several years.

The mammalian Ccr4–Not complex, carbon catabolite repression 4 (Ccr4)-negative on TATA-less (Not), is a large, highly conserved, multifunctional assembly of proteins that acts at different cellular levels to regulate gene expression. It is involved in the control of the cell cycle, chromatin modification, activation and inhibition of transcription initiation, control of transcription elongation, RNA export, and nuclear RNA surveillance; the Ccr4–Not complex also plays a central role in the regulation of mRNA decay. Growing evidence suggests that gene transcription has a vital role in shaping the landscape of genome replication and is also a potent source of replication stress and genome instability. Here, we have examined the effects of the inactivation of the Ccr4–Not complex, via the depletion of the scaffold subunit CNOT1, on DNA replication and genome integrity in mammalian cells. In CNOT1-depleted cells, the elevated expression of the general transcription factor TATA-box binding protein (TBP) leads to increased RNA synthesis, which, together with R-loop accumulation, results in replication fork slowing, DNA damage, and senescence. Furthermore, we have shown that the stability of TBP mRNA increases in the absence of CNOT1, which may explain its elevated protein expression in CNOT1-depleted cells. Finally, we have shown the activation of mitogen-activated protein kinase signalling as evidenced by ERK1/2 phosphorylation in the absence of CNOT1, which may be responsible for the observed cell cycle arrest at the border of G1/S.

Background: Meditation retreats are characterized by intensive or concentrated periods of meditation practice, commonly undertaken in a residential setting. Although research indicates that meditation training can positively influence physical and mental health outcomes, the biological consequences of meditation retreat interventions are relatively understudied. In this study, we examined the influence of a month-long, silent meditation retreat on the expression of genes involved in epigenetic modulation and immune processes. Method: We assessed gene expression changes in experienced meditators attending a month-long Insight meditation retreat (n = 28), as compared to a community control group (n = 34) of experienced practitioners living their everyday lives. Blood samples were collected on day two of the retreat (Time 1) and again 3 weeks later (Time 2). Control participants were also assessed across a 3-week interval, during which they maintained their regular daily routines. Results: As compared to controls, retreat participants showed differential changes in the expression of several genes involved in chromatin modulation and inflammation. The most substantive finding was downregulation of the TNF pathway in retreat participants, which was not observed in controls. Conclusions: These findings indicate that meditation retreat participation may influence some of the inflammatory mechanisms involved in the development of chronic diseases, and that this style of psychosocial intervention may have therapeutic potential, particularly in experienced practitioners.

Although blunted sensitivity to reward is thought to play a key role in promoting risk for depression, most research on this topic has utilized monetary reward paradigms and focused on currently depressed adults. To address this issue, we analyzed neural reward and β-endorphin data from the Psychobiology of Stress and Adolescent Depression (PSY SAD) Study, which recruited a well-characterized sample of adolescent girls at high vs. low risk for major depressive disorder (MDD) (N = 52, Mage = 14.90, SD = 1.35) based on their mothers’ lifetime history of MDD. As hypothesized, greater striatal activity while receiving positive (vs. neutral) social evaluation was associated with lower depression symptom severity as independently assessed by the Kiddie Schedule for Affective Disorders and Schizophrenia (K-SADS). This association was present for girls at high but not low risk for MDD, suggesting that this neural response may represent a pre-clinical marker of risk for depression. Consistent with these results, higher post-social evaluation levels of a peripheral marker of reward sensitivity, β-endorphin, were related to lower clinician-rated depression symptom severity. Together, these results indicate that neural and peripheral markers of responsivity to social reward are both related to depression severity, which may have implications for understanding the pathophysiology of depression.

Abstract Stress is a significant risk factor for the development of major depressive disorder (MDD), yet the underlying mechanisms remain unclear. Preclinically, adaptive and maladaptive stress-induced changes in glutamatergic function have been observed in the medial prefrontal cortex (mPFC). Here, we examine stress-induced changes in human mPFC glutamate using magnetic resonance spectroscopy (MRS) in two healthy control samples and a third sample of unmedicated participants with MDD who completed the Maastricht acute stress task, and one sample of healthy control participants who completed a no-stress control manipulation. In healthy controls, we find that the magnitude of mPFC glutamate response to the acute stressor decreases as individual levels of perceived stress increase. This adaptative glutamate response is absent in individuals with MDD and is associated with pessimistic expectations during a 1-month follow-up period. Together, this work shows evidence for glutamatergic adaptation to stress that is significantly disrupted in MDD.

Minichromosome maintenance protein 10 (MCM10) is critical for eukaryotic DNA replication. Here, by modelling MCM10 variants in human cell lines, the authors reveal a mechanism of MCM10-associated disease, finding that loss of MCM10 function constrains telomerase activity.

The feeding characteristics of the nase, based on its mouth morphology and feeding behavior related to aquatic habitat substrata sediments make this fish a biological uptake vector for microplastics in freshwater ecosystems. Fibers may have limited absorption through the gastrointestinal tract therefore unlikely to be found in fish gastrointestinal tissue and muscle tissue. The presence of microplastic fibers in the gastrointestinal content is proof of how difficult it is for these fibers to become embedded in other organs. The absence of microplastic fibers in muscle tissue and gastrointestinal tissue is key information for microplastic fabrication and management in aquatic ecosystems. The majority of fish have relatively low levels of microplastics; however, a few individuals have a higher dose. This is true for all types of microplastics analyzed, with the exception of fish that had just one microplastic present in the analyzed matrix. The microplastics are not concentrated in the fish muscle tissue, gastrointestinal tissue and gastrointestinal content in relation to fish age, which may be due to their different mobility in the ecosystem, or due to the recent appearance of this type of contamination and the scale at which microplastics bioaccumulate. Such a relatively common fish species must be included in the assessment and monitoring systems of the Eurasian lotic systems. The risks involved include the transfer of the freshwater environments microplastics into human tissues via the food web of fishing species with the nase as a key basis.

Abstract Background More than 75% of patients presenting to the Emergency Department are suffering symptoms of pain. Despite this, 67% will not receive any analgesia. Methoxyflurane is a fluorinated hydrocarbon gas which has analgesic properties when inhaled. Penthrox is a methoxyflurane autoinhaler recently licenced in Europe. Its ease of administration, safety, and fast onset of action make it of particular relevance to emergency medicine. Additionally, outside the hospital, it has the advantage of increased temperature stability and portability over current standard care. New evidence of its efficacy is emerging; however, currently, its use in Europe is not widespread. The objective of this study will be to systematically evaluate the evidence on inhaled methoxyflurane to determine if it is a superior analgesia in the acute trauma setting. Methods We designed and registered a study protocol for a systematic review and meta-analysis on randomised controlled trials, comparing inhaled methoxyflurane and either placebo or standard care. A comprehensive search will be conducted from database inception onwards in MEDLINE, Embase, and the Cochrane CENTRAL database, concurrent with a search of the grey literature for other relevant studies, including clinical trial databases. Only randomised controlled trials will be included. No limitations will be imposed on publication status or language of publication. The primary outcome will be mean difference in patient-reported pain at time points within the first 30 min of administration. Secondary outcomes will be mean difference in time to clinically significant pain relief and relative risk of adverse effects. Two reviewers will independently screen all returned studies and collect data. Disagreements will be resolved through discussion or referral to a third reviewer. Individual study methodological quality will be appraised using an appropriate tool. If feasible, we will conduct a random effects meta-analysis; if this is not possible, we will construct a narrative synthesis. Discussion This systematic review will summarise the best available evidence and definitively establish if inhaled methoxyflurane is a superior analgesia to standard care in the acute trauma setting. This knowledge will directly impact emergency care in the UK and worldwide and may require amendments to European pain relief guidelines. Systematic review registration PROSPERO CRD42020189119 .

Abstract High-altitude-adapted ectotherms can escape competition from dominant species by tolerating low temperatures at cooler elevations, but climate change is eroding such advantages. Studies evaluating broad-scale impacts of global change for high-altitude organisms often overlook the mitigating role of biotic factors. Yet, at fine spatial-scales, vegetation-associated microclimates provide refuges from climatic extremes. Using one of the largest standardised data sets collected to date, we tested how ant species composition and functional diversity (i.e., the range and value of species traits found within assemblages) respond to large-scale abiotic factors (altitude, aspect), and fine-scale factors (vegetation, soil structure) along an elevational gradient in tropical Africa. Altitude emerged as the principal factor explaining species composition. Analysis of nestedness and turnover components of beta diversity indicated that ant assemblages are specific to each elevation, so species are not filtered out but replaced with new species as elevation increases. Similarity of assemblages over time (assessed using beta decay) did not change significantly at low and mid elevations but declined at the highest elevations. Assemblages also differed between northern and southern mountain aspects, although at highest elevations, composition was restricted to a set of species found on both aspects. Functional diversity was not explained by large scale variables like elevation, but by factors associated with elevation that operate at fine scales (i.e., temperature and habitat structure). Our findings highlight the significance of fine-scale variables in predicting organisms’ responses to changing temperature, offering management possibilities that might dilute climate change impacts, and caution when predicting assemblage responses using climate models, alone.

Stem cells could form the basis of a novel, autologous treatment for chronic wounds like diabetic foot ulcers. Fat grafts contain adipose-derived stem cells (ADSC) but low survival of cells within the grafts is a major limitation. Platelet-rich plasma (PRP) may increase graft survival. This review examines the histology from animal studies on fat grafting, ADSC and PRP in wound healing. A literature review of major electronic databases was undertaken, and narrative synthesis performed. Data from 30 animal studies were included. ADSC increase angiogenesis over 14 days and often clinically accelerated wound healing. ADSC had a greater effect in animals with impaired wound healing (e.g. diabetes). Activated PRP increased viability of fat grafts. Despite the high number of studies, the quality is variable which weakens the evidence. It does suggest there is a benefit of ADSC, particularly in impaired wound healing. High-quality evidence in humans is required, to establish its clinical usefulness.

Despite being the first homolog of the bacterial RecQ helicase to be identified in humans, the function of RECQL1 remains poorly characterized. Furthermore, unlike other members of the human RECQ family of helicases, mutations in RECQL1 have not been associated with a genetic disease. Here, we identify 2 families with a genome instability disorder that we have named RECON (RECql ONe) syndrome, caused by biallelic mutations in the RECQL gene. The affected individuals had short stature, progeroid facial features, a hypoplastic nose, xeroderma, and skin photosensitivity and were homozygous for the same missense mutation in RECQL1 (p.Ala459Ser), located within its zinc binding domain. Biochemical analysis of the mutant RECQL1 protein revealed that the p.A459S missense mutation compromised its ATPase, helicase, and fork restoration activity, while its capacity to promote single-strand DNA annealing was largely unaffected. At the cellular level, this mutation in RECQL1 gave rise to a defect in the ability to repair DNA damage induced by exposure to topoisomerase poisons and a failure of DNA replication to progress efficiently in the presence of abortive topoisomerase lesions. Taken together, RECQL1 is the fourth member of the RecQ family of helicases to be associated with a human genome instability disorder.