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Abstract Aims Hyperactivity of Ca2+/calmodulin‐dependent protein kinase II (CaMKII) has emerged as a central cause of pathologic remodelling in heart failure. It has been suggested that CaMKII‐induced hyperphosphorylation of the ryanodine receptor 2 (RyR2) and consequently increased diastolic Ca2+ leak from the sarcoplasmic reticulum (SR) is a crucial mechanism by which increased CaMKII activity leads to contractile dysfunction. We aim to evaluate the relevance of CaMKII‐dependent RyR2 phosphorylation for CaMKII‐induced heart failure development in vivo. Methods and results We crossbred CaMKIIδC overexpressing [transgenic (TG)] mice with RyR2‐S2814A knock‐in mice that are resistant to CaMKII‐dependent RyR2 phosphorylation. Ca2+‐spark measurements on isolated ventricular myocytes confirmed the severe diastolic SR Ca2+ leak previously reported in CaMKIIδC TG [4.65 ± 0.73 mF/F0 vs. 1.88 ± 0.30 mF/F0 in wild type (WT)]. Crossing in the S2814A mutation completely prevented SR Ca2+‐leak induction in the CaMKIIδC TG, both regarding Ca2+‐spark size and frequency, demonstrating that the CaMKIIδC‐induced SR Ca2+ leak entirely depends on the CaMKII‐specific RyR2‐S2814 phosphorylation. Yet, the RyR2‐S2814A mutation did not affect the massive contractile dysfunction (ejection fraction = 12.17 ± 2.05% vs. 45.15 ± 3.46% in WT), cardiac hypertrophy (heart weight/tibia length = 24.84 ± 3.00 vs. 9.81 ± 0.50 mg/mm in WT), or severe premature mortality (median survival of 12 weeks) associated with cardiac CaMKIIδC overexpression. In the face of a prevented SR Ca2+ leak, the phosphorylation status of other critical CaMKII downstream targets that can drive heart failure, including transcriptional regulator histone deacetylase 4, as well as markers of pathological gene expression including Xirp2, Il6, and Col1a1, was equally increased in hearts from CaMKIIδC TG on a RyR WT and S2814A background. Conclusions S2814 phosphoresistance of RyR2 prevents the CaMKII‐dependent SR Ca2+ leak induction but does not prevent the cardiomyopathic phenotype caused by enhanced CaMKIIδC activity. Our data indicate that additional mechanisms—independent of SR Ca2+ leak—are critical for the maladaptive effects of chronically increased CaMKIIδC activity with respect to heart failure.

Abstract Distinguishing the independent contributions of tectonic denudation and climate‐driven erosion on the production and supply of clastic materials of Chinese eolian deposits is important in understanding the dynamic links between global climate changes, tectonics, and Asian dust emission. Here, multi‐proxy rock magnetic records of detrital fractions of Chinese eolian deposits since ∼6 Ma, combined with further comparison with dust accumulation rates and geochemical data, suggest that limited production and supply of clastic materials of Chinese eolian deposits were observed between 5.6 and 4.4 Ma in a warmer climate, and thereafter an increasing trend is evident during a cooler world. We propose that intense physical erosion under cooler conditions caused increasing production of fresh detrital components, exerting dominant influences on the sediment supply to Asian eolian deposits. This study provides direct evidence of dynamic coupling between intensified physical erosion and Asian dust activity under late Cenozoic global cooling.

Prostate cancer is a frequent malignancy in older men and has a very high 5‐year survival rate if diagnosed early. The prognosis is much less promising if the tumor has already spread outside the prostate gland. Targeted treatments mainly aim at blocking androgen receptor (AR) signaling and initially show good efficacy. However, tumor progression due to AR‐dependent and AR‐independent mechanisms is often observed after some time, and novel treatment strategies are urgently needed. Dysregulation of the PI3K/AKT/mTOR pathway in advanced prostate cancer and its implication in treatment resistance has been reported. We compared the impact of PI3K/AKT/mTOR pathway inhibitors with different selectivity profiles on in vitro cell proliferation and on caspase 3/7 activation as a marker for apoptosis induction, and observed the strongest effects in the androgen‐sensitive prostate cancer cell lines VCaP and LNCaP. Combination treatment with the AR inhibitor darolutamide led to enhanced apoptosis in these cell lines, the effects being most pronounced upon cotreatment with the pan‐PI3K inhibitor copanlisib. A subsequent transcriptomic analysis performed in VCaP cells revealed that combining darolutamide with copanlisib impacted gene expression much more than individual treatment. A comprehensive reversal of the androgen response and the mTORC1 transcriptional programs as well as a marked induction of DNA damage was observed. Next, an in vivo efficacy study was performed using the androgen‐sensitive patient‐derived prostate cancer (PDX) model LuCaP 35 and a superior efficacy was observed after the combined treatment with copanlisib and darolutamide. Importantly, immunohistochemistry analysis of these treated tumors showed increased apoptosis, as revealed by elevated levels of cleaved caspase 3 and Bcl‐2‐binding component 3 (BBC3). In conclusion, these data demonstrate that concurrent blockade of the PI3K/AKT/mTOR and AR pathways has superior antitumor efficacy and induces apoptosis in androgen‐sensitive prostate cancer cell lines and PDX models.

Abstract Pulmonary arterial hypertension (PAH) is characterized by progressive pulmonary vascular remodeling with resultant abnormal increase in pulmonary artery pressure and right heart dysfunction. There is evidence that PAH includes cognitive impairment. However, the cognitive impairment syndrome has not been well described, and both the underlying mechanism and the relationship between cardiopulmonary and cognitive dysfunction in PAH are unknown. We performed cognitive evaluations and same day sub‐maximum cardiopulmonary exercise testing on adult subjects with PAH. A frontal–subcortical syndrome suggestive of vascular cognitive impairment was found in 26% of subjects and was associated with noninvasive markers of pulmonary vascular remodeling.

Abstract Background Blood transfusions are performed frequently in goats, but crossmatches are rarely performed. Hypothesis/Objectives Determine differences in the frequency of agglutination and hemolytic crossmatch reactions between large and small breed goats. Animals Healthy adult goats, 10 large and 10 small breed. Methods Two hundred eighty major and minor agglutination and hemolytic crossmatches: 90 large breed donor to large breed recipient (L‐L), 90 small breed donor to small breed recipient (S‐S), 100 large breed donor to small breed recipient (L‐S). A linear mixed model with treatment group (L‐L, S‐S, L‐S) as a fixed effect and individual crossmatch as a random effect was used to identify variations in reaction frequency among groups and individuals. Results Frequency of major agglutination reactions for L‐L, S‐S, and L‐S were 3/90 (3.3%), 7/90 (7.8%), and 10/100 (10.0%), respectively. Frequency of major hemolytic reactions for L‐L, S‐S, and L‐S were 27/84 (32.1%), 7/72 (9.7%), and 31/71 (43.7%). Individual pairings and groupings had no effect on agglutination reactions. Individual pairings had no effect on the frequency of hemolytic reactions. For major hemolytic crossmatches, pairwise comparisons identified higher frequencies of reactions when comparing L‐L to S‐S (P = .007) and L‐S to S‐S (P

Abstract Hydroxychloroquine (HCQ) is Food and Drug Administration (FDA)‐approved for malaria, systemic and chronic discoid lupus erythematosus, and rheumatoid arthritis. Because HCQ has a proposed multimodal mechanism of action and a well‐established safety profile, it is often investigated as a repurposed therapeutic for a range of indications. There is a large degree of uncertainty in HCQ pharmacokinetic (PK) parameters which complicates dose selection when investigating its use in new disease states. Complications with HCQ dose selection emerged as multiple clinical trials investigated HCQ as a potential therapeutic in the early stages of the COVID‐19 pandemic. In addition to uncertainty in baseline HCQ PK parameters, it was not clear if disease‐related consequences of SARS‐CoV‐2 infection/COVID‐19 would be expected to impact the PK of HCQ and its primary metabolite desethylhydroxychloroquine (DHCQ). To address the question whether SARS‐CoV‐2 infection/COVID‐19 impacted HCQ and DHCQ PK, dried blood spot samples were collected from SARS‐CoV‐2(−)/(+) participants administered HCQ. When a previously published physiologically based pharmacokinetic (PBPK) model was used to fit the data, the variability in exposure of HCQ and DHCQ was not adequately captured and DHCQ concentrations were overestimated. Improvements to the previous PBPK model were made by incorporating the known range of blood to plasma concentration ratios (B/P) for each compound, adjusting HCQ and DHCQ distribution settings, and optimizing DHCQ clearance. The final PBPK model adequately captured the HCQ and DHCQ concentrations observed in SARS‐CoV‐2(−)/(+)participants, and incorporating COVID‐19‐associated changes in cytochrome P450 activity did not further improve model performance for the SARS‐CoV‐2(+) population.

Abstract The use of arbuscular mycorrhizal fungi (AMF) as biofertilizers has proven successful in boosting the yield and nutritional quality of a variety of crops. AMF associate with plant roots and exchange soil nutrients for photosynthetically derived C in the form of sugars and lipids. Past research has shown that not all AMF species are equal in their benefit to nutrient uptake and crop health, and that the most beneficial AMF species appear to vary by host species. Although an important human food staple, especially in developing regions where nutrient deficiency is a prevalent threat to public health, little work has been done to test the effectiveness of AMF in enhancing the nutritional quality of common bean (Phaseolus vulgaris L.). Therefore, our objective was to determine the most beneficial AMF species for inoculation of this important crop. We inoculated black beans (Phaseolus vulgaris black turtle beans) with eight individual AMF species and one mixed species inoculum in an outdoor pot trial over 3 months and assessed the extent to which they altered yield, mineral nutrient and anthocyanin concentration of seeds and leaf tissues. Despite seeing no yield effects from inoculation, we found that across treatments percent root length colonized by AMF was positively correlated with plant tissue P, Cu, and Zn concentration. Underlying these broad benefits, seeds from plants inoculated with three AMF species, Claroideoglomus claroideum (+15%), Funneliformis mosseae (+13%), and Gigaspora rosea (+11%) had higher P concentration than non‐mycorrhizal plants. C. claroideum also increased seed potassium (K) and copper (Cu), as well as leaf aluminum (Al) concentration making it a promising candidate to further test the benefit of individual AMF species on black bean growth in field trials.

Abstract Objectives Dominant‐activating (DA) lesions in RAC2 have been reported in 18 individuals to date. Some have required haematopoietic stem cell transplantation (HSCT) for their (severe) combined immunodeficiency syndrome phenotype. We aimed to investigate clinical and cellular features of a kindred harbouring a novel variant in RAC2 p.Ile21Ser (I21S) to better understand DA lesions' phenotypic spectrum. Methods Clinical and immunological information was collated for seven living individuals from the same kindred with RAC2 p.I21S. We evaluated neutrophil morphology, RAC2 protein expression and superoxide production using freshly isolated neutrophils stimulated with phorbol‐12‐myristate‐13‐acetate (PMA) and N‐formyl‐MetLeuPhe (fMLP). Results Patient 1 (P1, aged 11, male) has a history of bacterial suppurative otitis media, viral and bacterial cutaneous infections. P1's siblings (P2, P3), mother (P4), maternal aunt (P5) and uncle (P6) have similar infection histories. P1's maternal cousin (P7) presented with Burkitt's lymphoma at age 9. All affected individuals are alive and none has required HSCT to date. They have chronic lymphopenia affecting the CD4+T and B‐cell compartments. P1–3 have isolated reduction in IgM levels whereas the adults universally have normal immunoglobulins. Specific antibody responses are preserved. Affected individuals have neutrophil vacuolation, and their neutrophils have enhanced superoxide production compared to healthy controls. Conclusion RAC2 p.I21S is an activating variant causing notable morphological and functional abnormalities similar to other reported DA mutations. This novel variant expands the broad clinical phenotypic spectrum of RAC2 DA lesions, emphasising the need to tailor clinical management according to patients' disease phenotype and severity.

Societal Impact Statement Evidence has emerged that the antioxidant ergothioneine may be important in preventing many inflammatory diseases in humans. However, ergothioneine is not produced by humans or plants and is only made by fungi and some bacteria in soils. As such, humans get ergothioneine from eating fungi (mushrooms) or plants that take it up from the soil. In this study, we found that growing plants with beneficial fungi called arbuscular mycorrhizal fungi increased the amount of ergothioneine in plant tissues. This suggests that promoting agricultural practices that maintain healthy populations of beneficial soil fungi may improve the nutritional quality of crops. Summary The amino acid ergothioneine (ERGO) has recently gained attention as an antioxidant that benefits human health. ERGO is produced by fungi and mycobacteria in soils and is acquired only from diet. The mechanism by which ERGO is transferred from soil to plant is unknown. Recent work has shown that tillage reduces the amount of ERGO in crops. As tillage also reduces arbuscular mycorrhizal fungi (AMF) populations, we examined the relationship between AMF and plant ERGO uptake. We grew asparagus, black beans, wheat, and oats with a variety of single and mixed species AMF inocula and compared ERGO levels of these plants to plants that were uninoculated. Mycorrhizal inoculation enhanced ERGO content across all plants. There was a positive correlation between AMF colonization level and plant ERGO content. AMF appear to be important mediators of plant ERGO uptake. Future research is needed to identify the mechanism that leads to higher ERGO in plants colonized by AMF in order to promote farming practices that enhance AMF populations and increase crop ERGO concentration in field settings.

Abstract Objective Body volumes (BV) are used for calculating body composition to perform obesity assessments. Conventional BV estimation techniques, such as underwater weighing, can be difficult to apply. Advanced machine learning techniques enable multiple obesity‐related body measurements to be obtained using a single‐camera image; however, the accuracy of BV calculated using these techniques is unknown. This study aims to adapt and evaluate a machine learning technique, synthetic training for real accurate pose and shape (STRAPS), to estimate BV. Methods The machine learning technique, STRAPS, was applied to generate three‐dimensional (3D) models from simulated two‐dimensional (2D) images; these 3D models were then scaled with body stature and BV were estimated using regression models corrected for body mass. A commercial 3D scan dataset with a wide range of participants (n = 4318) was used to compare reference and estimated BV data. Results The developed methods estimated BV with small relative standard errors of estimation (

Abstract The first step in phage infection is the recognition of, and adsorption to, a receptor located on the host cell surface. This reversible host adsorption step is commonly followed by an irreversible event, which involves phage DNA delivery or release into the bacterial cytoplasm. The molecular components that trigger this latter event are unknown for most phages of Gram‐positive bacteria. In the current study, we present a comparative genome analysis of three mutants of Lactococcus cremoris 3107, which are resistant to the P335 group phage TP901‐1 due to mutations that affect TP901‐1 DNA release. Through genetic complementation and phage infection assays, a predicted lactococcal three‐component glycosylation system (TGS) was shown to be required for TP901‐1 infection. Major cell wall saccharidic components were analysed, but no differences were found. However, heterologous gene expression experiments indicate that this TGS is involved in the glucosylation of a cell envelope‐associated component that triggers TP901‐1 DNA release. To date, a saccharide modification has not been implicated in the DNA delivery process of a Gram‐positive infecting phage.

Abstract Background Neuroinflammation has been shown to be an important pathophysiological disease mechanism in frontotemporal dementia (FTD). This includes activation of microglia, a process that can be measured in life through assaying different glia‐derived biomarkers in cerebrospinal fluid. However, only a few studies so far have taken place in FTD, and even fewer focusing on the genetic forms of FTD. Methods We investigated the cerebrospinal fluid concentrations of TREM2, YKL‐40 and chitotriosidase using immunoassays in 183 participants from the Genetic FTD Initiative (GENFI) study: 49 C9orf72 (36 presymptomatic, 13 symptomatic), 49 GRN (37 presymptomatic, 12 symptomatic) and 23 MAPT (16 presymptomatic, 7 symptomatic) mutation carriers and 62 mutation‐negative controls. Concentrations were compared between groups using a linear regression model adjusting for age and sex, with 95% bias‐corrected bootstrapped confidence intervals. Concentrations in each group were correlated with the Mini‐Mental State Examination (MMSE) score using non‐parametric partial correlations adjusting for age. Age‐adjusted z‐scores were also created for the concentration of markers in each participant, investigating how many had a value above the 95th percentile of controls. Results Only chitotriosidase in symptomatic GRN mutation carriers had a concentration significantly higher than controls. No group had higher TREM2 or YKL‐40 concentrations than controls after adjusting for age and sex. There was a significant negative correlation of chitotriosidase concentration with MMSE in presymptomatic GRN mutation carriers. In the symptomatic groups, for TREM2 31% of C9orf72, 25% of GRN, and 14% of MAPT mutation carriers had a concentration above the 95th percentile of controls. For YKL‐40 this was 8% C9orf72, 8% GRN and 0% MAPT mutation carriers, whilst for chitotriosidase it was 23% C9orf72, 50% GRN, and 29% MAPT mutation carriers. Conclusions Although chitotriosidase concentrations in GRN mutation carriers were the only significantly raised glia‐derived biomarker as a group, a subset of mutation carriers in all three groups, particularly for chitotriosidase and TREM2, had elevated concentrations. Further work is required to understand the variability in concentrations and the extent of neuroinflammation across the genetic forms of FTD. However, the current findings suggest limited utility of these measures in forthcoming trials.

Abstract Eolian loess deposits contain valuable information about past climate changes and erosion history in dust source regions. In contrast to the extensive investigations of paleoclimatic implications of Eurasian loess, continental erosion information in the loess deposits has received less attention, particularly on the global or hemispheric scales. The present study combines citrate‐bicarbonate‐dithionite (CBD) magnetic mineral extraction procedures and linear regression of magnetic parameters to analyze Eurasian loess. We find that lithogenic susceptibility of Quaternary loess deposits in Central Asia, Europe and the Chinese Loess Plateau shows synchronous long‐term increases since ∼0.6–0.5 Ma, suggesting intensive glacial erosion and/or river incision to have occurred in the surrounding mountains of dust source regions after the mid‐Pleistocene climate Transition. The dramatic increase in lithogenic susceptibility of Eurasian loess provides new insights into the close relationships between global climate changes and dust source erosion since the late Cenozoic.

Abstract We have modeled the high‐frequency seismogram envelopes of the large event S1222a and four recently identified near impacts recorded by the InSight mission by introducing a stratification of velocity and attenuation into a multiple‐scattering approach. We show that a simple conceptual model composed of a strongly diffusive, weakly attenuating layer overlying a transparent medium captures the essential features of the observed envelopes. The attenuation profiles reveal that the minimal extension of heterogeneities at depth is of the order of 20 km in the vicinity of InSight and 60 km on the path to S1222a. We interpret this result as an indication that the Martian crust as a whole is at the origin of the strong scattering. Our heterogeneity model suggests that the sources of a number of distant Very‐high‐Frequency seismic events are shallow and located to the south or in close vicinity of the Martian dichotomy.

Uveal melanoma (UM) is the most common intraocular tumor in adults. Recurrent mutations in BRCA1‐associated protein 1 (BAP1) and splicing factor 3B subunit 1 (SF3B1) display a mutually exclusive pattern in UM, but the underlying mechanism is unknown. We show that combined BAP1 deficiency and SF3B1 hotspot mutation lead to senescence and growth arrest in human UM cells. Although p53 protein expression is induced, deletion of TP53 (encoding p53) only modestly rescues the observed senescent phenotype. UM cells with BAP1 loss or SF3B1 mutation are more sensitive to chemotherapeutic drugs compared with their isogenic parental cells. Transcriptome analysis shows that DNA‐repair genes are downregulated upon co‐occurrence of BAP1 deletion and SF3B1 mutation, thus leading to impaired DNA damage response and the induction of senescence. The co‐occurrence of these two mutations reduces invasion of UM cells in zebrafish xenograft models and suppresses growth of melanoma xenografts in nude mice. Our findings provide a mechanistic explanation for the mutual exclusivity of BAP1 and SF3B1 mutations in human UM.

Background Peanut allergy is a frequent cause of food allergy and potentially life‐threatening. Within this interdisciplinary research approach, we aim to unravel the complex mechanisms of peanut allergy. As a first step were applied in an exploratory manner the analysis of peanut allergic versus non‐allergic controls. Methods Biosamples were studied regarding DNA methylation signatures, gut microbiome, adaptive and innate immune cell populations, soluble signaling molecules and allergen‐reactive antibody specificities. We applied a scalable systems medicine computational workflow to the assembled data. Results We identified combined cellular and soluble biomarker signatures that stratify donors into peanut‐allergic and non‐allergic with high specificity. DNA methylation profiling revealed various genes of interest and stool microbiota differences in bacteria abundances. Conclusion By extending our findings to a larger set of patients (e.g., children vs. adults), we will establish predictors for food allergy and tolerance and translate these as for example, indicators for interventional studies.

Abstract Background In dogs, antiepileptic drugs (AED) cause lethargy but quantitative data regarding the effects of AED on activity levels are not available, and little is known about how AEDs affect sleep quality. Objective To quantitatively compare activity levels and nocturnal activity in dogs previously diagnosed with idiopathic epilepsy (IE) receiving AEDs compared to age‐ and breed‐matched control dogs. Animals Sixty‐two dogs with IE and 310 control dogs. Methods This is a 3‐month prospective parallel observational study. An activity monitoring device for dogs was used to measure daily activity levels and sleep scores in all dogs. Results Dogs with IE treated with AEDs had an 18% average lower baseline activity level compared to control dogs (P = .005; point estimate = 0.82, 95% confidence interval [CI], 0.75‐0.90). The combination of phenobarbital and potassium bromide (KBr) was associated with an average 28% decrease in activity in dogs with IE compared to control dogs (P = .03; point estimate = 0.72; CI, 0.62‐0.82). Mean sleep scores were not significantly different in dogs with IE receiving AEDs compared to control dogs (P = .43). However, higher dosages of KBr were associated with lower sleep scores (P = .01). Conclusions Dogs with IE receiving AEDs have lower activity levels, but no difference in sleep scores, compared to controls. The combination of phenobarbital and KBr had the largest decrease in activity between groups. Higher doses of KBr may affect nocturnal activity in epileptic dogs.

Abstract Four, mature, client‐owned goats were presented to referral hospitals for recurrent diarrhea despite treatment for intestinal parasitism. Common clinical findings included diarrhea, poor condition, neutrophilia, and hypoalbuminemia. Testing for common infectious causes of diarrhea in goats was negative. Ultrasonography and computed tomography in 2 cases was suggestive of enteritis, including thickened intestinal walls and fluid filled, dilated small intestines, respectively. Lymphoplasmacytic and eosinophilic enteritis (LEE) was ultimately diagnosed on intestinal biopsy histopathology based on the presence of small intestinal villous blunting and increased numbers of lymphocytes and eosinophils predominantly within the lamina propria. Numerous globule leukocytes were also noted on histopathology in 3 cases. All goats responded favorably to corticosteroid treatment with weight gain and resolution of diarrhea and clinicopathologic abnormalities. Relapses occurred, and complete cure was difficult to achieve. Reported in other species, this series describes the clinical presentation, diagnosis, and treatment of LEE in adult goats.

Abstract Aims White matter lesions (WML) are common structural alterations in the white matter of the brain and their prevalence increases with age. They are associated with cerebral ischaemia and cognitive dysfunction. Patients with heart failure (HF) are at risk for cognitive decline. We hypothesized that the presence and duration of HF are associated with WML. Methods and results The LIFE‐Adult Study is a population‐based study of 10 000 residents of Leipzig, Germany. WML were quantitated in 2490 participants who additionally underwent cerebral MRI using the Fazekas score. Mean age was 64 years, and 46% were female; 2156 (86.6%) subjects had Fazekas score of 0–1, and 334 (13.4%) had Fazekas score of 2–3. Thirty participants had a medical history of HF, 1019 had hypertension, and 51 had a history of stroke. Median left ventricular ejection fraction of the participants with HF was 57% (interquartile ranges 54–62). Age, troponin T, NT‐proBNP, body mass index, history of acute myocardial infarction, stroke, HF, and diabetes were positively associated with WML in univariate analysis. On multivariate analysis, age, hypertension, stroke, and HF were independently associated with WML. The odd's ratio for the association of WML (Fazekas 2–3) with HF was 2.8 (95% CI 1.2–6.5; P = 0.019). WML increased with longer duration of HF (P = 0.036 for trend). Conclusions In addition to age, hypertension, and stroke, the prevalence and duration of HF are independently associated with WML. This observation sets the stage to investigate the prognostic value of WML in HF and the impact of HF therapies on WML.