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Abstract Owing to its roles in cellular signal transduction, protein phosphorylation plays critical roles in myriad cell processes. That said, detecting and quantifying protein phosphorylation has remained a challenge. We describe the use of a novel mass spectrometer (Orbitrap Astral) coupled with data-independent acquisition (DIA) to achieve rapid and deep analysis of human and mouse phosphoproteomes. With this method, we map approximately 30,000 unique human phosphorylation sites within a half-hour of data collection. The technology is benchmarked to other state-of-the-art MS platforms using both synthetic peptide standards and with EGF-stimulated HeLa cells. We apply this approach to generate a phosphoproteome multi-tissue atlas of the mouse. Altogether, we detect 81,120 unique phosphorylation sites within 12 hours of measurement. With this unique dataset, we examine the sequence, structural, and kinase specificity context of protein phosphorylation. Finally, we highlight the discovery potential of this resource with multiple examples of phosphorylation events relevant to mitochondrial and brain biology.

A new muon beam line, the muon science innovative channel, was set up at the Research Center for Nuclear Physics, Osaka University, in Osaka, Japan, using the 392 MeV proton beam impinging on a target. The production of an intense muon beam relies on the efficient capture of pions, which subsequently decay to muons, using a novel superconducting solenoid magnet system. After the pion-capture solenoid, the first 36° of the curved muon transport line was commissioned and the muon flux was measured. In order to detect muons, a target of either copper or magnesium was placed to stop muons at the end of the muon beam line. Two stations of plastic scintillators located upstream and downstream from the muon target were used to reconstruct the decay spectrum of muons. In a complementary method to detect negatively charged muons, the x-ray spectrum yielded by muonic atoms in the target was measured in a germanium detector. Measurements, at a proton beam current of 6 pA, yielded (10.4±2.7)×10^{5} muons per watt of proton beam power (μ^{+} and μ^{-}), far in excess of other facilities. At full beam power (400 W), this implies a rate of muons of (4.2±1.1)×10^{8} muons s^{−1}, among the highest in the world. The number of μ^{-} measured was about a factor of 10 lower, again by far the most efficient muon beam produced. The setup is a prototype for future experiments requiring a high-intensity muon beam, such as a muon collider or neutrino factory, or the search for rare muon decays which would be a signature for phenomena beyond the Standard Model of particle physics. Such a muon beam can also be used in other branches of physics, nuclear and condensed matter, as well as other areas of scientific research.

Abstract AKS-452, a subunit vaccine comprising an Fc fusion of the ancestral wild-type (WT) SARS-CoV-2 virus spike protein receptor binding domain (SP/RBD), was evaluated without adjuvant in a single cohort, non-randomized, open-labelled phase II study (NCT05124483) at a single site in The Netherlands for safety and immunogenicity. A single 90 µg subcutaneous booster dose of AKS-452 was administered to 71 adults previously primed with a registered mRNA- or adenovirus-based vaccine and evaluated for 273 days. All AEs were mild and no SAEs were attributable to AKS-452. While all subjects showed pre-existing SP/RBD binding and ACE2-inhibitory IgG titers, 60–68% responded to AKS-452 via ≥2-fold increase from days 28 to 90 and progressively decreased back to baseline by day 180 (days 28 and 90 mean fold-increases, 14.7 ± 6.3 and 8.0 ± 2.2). Similar response kinetics against RBD mutant proteins (including omicrons) were observed but with slightly reduced titers relative to WT. There was an expected strong inverse correlation between day-0 titers and the fold-increase in titers at day 28. AKS-452 enhanced neutralization potency against live virus, consistent with IgG titers. Nucleocapsid protein (Np) titers suggested infection occurred in 66% (46 of 70) of subjects, in which only 20 reported mild symptomatic COVID-19. These favorable safety and immunogenicity profiles support booster evaluation in a planned phase III universal booster study of this room-temperature stable vaccine that can be rapidly and inexpensively manufactured to serve vaccination at a global scale without the need of a complex distribution or cold chain.

In this work, we optically resolve and detect individual rubidium atoms trapped in solid neon. Additionally, we optically pump the rubidium's spin state using polarized light and measure the spin state via laser-induced fluorescence. When combined with the previously demonstrated magnetic field sensing capabilities of matrix-isolated rubidium atoms, these results are very promising for nanoscale sensing and for performing nuclear magnetic resonance (NMR) spectroscopy of individual molecules cotrapped in the matrix.

IntroductionThe antigen-presenting cell function of insulin-reactive B cells promotes type 1 diabetes (T1D) in non-obese diabetic (NOD) mice by stimulating pathogenic T cells leading to destruction of insulin-producing β-cells of pancreatic islets.Methods/ResultsTo target insulin-reactive B cells, AKS-107, a human IgG1 Fc molecule fused with human insulin A and B chains, was engineered to retain conformational insulin epitopes that bound mouse and human B cell receptors but prevented binding to the insulin metabolic receptor. AKS-107 Fc-mediated deletion of insulin-reactive B cells was demonstrated via ex vivo and in vivo experiments with insulin-reactive B cell receptor transgenic mouse strains, VH125Tg/NOD and Tg125(H+L)/NOD. As an additional immune tolerance feature, the Y16A mutation of the insulin B(9-23) dominant T cell epitope was engineered into AKS-107 to suppress activation of insulin-specific T cells. In mice and non-human primates, AKS-107 was well-tolerated, non-immunogenic, did not cause hypoglycemia even at high doses, and showed an expectedly protracted pharmacokinetic profile. AKS-107 reproducibly prevented spontaneous diabetes from developing in NOD and VH125Tg/NOD mice that persisted for months after cessation of treatment, demonstrating durable immune tolerance.DiscussionThese preclinical outcomes position AKS-107 for clinical development in T1D prevention settings.

Improving the shelf life of steaks from beef chuck rolls and bone-in short ribs, items commonly exported, will increase the demand of beef in international markets. Our objectives were to determine the effect of topically applying combinations of acerola cherry powder and rosemary extract on steak color and lipid oxidation of beef chuck roll and bone-in short rib. USDA Choice beef chuck rolls (Institutional Meat Purchase Specifications [IMPS] 116A; N=9) andbone-in short ribs (IMPS 123A; N=18) were aged (0°C) 28 d and cut into 1.0 cm-thick steaks. Steak treatments included the following: untreated control (C) or topically sprayed (2 mL) with an acerola cherry powder solution (0.05% A), rosemary extract solution (0.10% R), or mixture of acerola cherry powder and rosemary extract (M1=0.05% A+0.1% R; M2=0.1% A+0.1% R; M3=0.05% A+0.2% R; M4=0.1% A+0.2% R). On day 0 of retail display, oxygen consumption, metmyoglobin-reducing activity, and lipid oxidation were measured. Lipid oxidation was evaluated again following 4 d of retail display. Chuck roll steak color was scored twice daily, whereas short rib lean and bone marrow color were scored once daily throughout 4 d of retail display. Treating chuck roll steaks with M2, M3, and M4 decreased lipid oxidation compared to C and A (P=0.004). In short rib steaks, lipid oxidation remained constant from day 0 to 4 treated with M4 but increased for all other treatments (P

Abstract When molecular testing classifies breast tumors as low risk but clinical risk is high, the optimal management strategy is unknown. One group of patients who may be more likely to have such discordant risk are those with invasive lobular carcinoma of the breast. We sought to examine whether patients with invasive lobular carcinoma are more likely to have clinical high/genomic low-risk tumors compared to those with invasive ductal carcinoma, and to evaluate the impact on receipt of chemotherapy and overall survival. We conducted a cohort study using the National Cancer Database from 2010–2016. Patients with hormone receptor positive, HER2 negative, stage I-III breast cancer who underwent 70-gene signature testing were included. We evaluated the proportion of patients with discordant clinical and genomic risk by histology using Kaplan-Meier plots, log-rank tests, and Cox proportional hazards models with and without propensity score matching. A total of 7399 patients (1497 with invasive lobular carcinoma [20.2%]) were identified. Patients with invasive lobular carcinoma were significantly more likely to fall into a discordant risk category compared to those with invasive ductal carcinoma (46.8% versus 37.1%, p

The parahippocampal cingulum bundle (PHCB) interconnects regions known to be vulnerable to early Alzheimer's disease (AD) pathology, including posteromedial cortex and medial temporal lobe. While AD-related pathology has been robustly associated with alterations in PHCB microstructure, specifically lower fractional anisotropy (FA) and higher mean diffusivity (MD), emerging evidence indicates that the reverse pattern is evident in younger adults at increased risk of AD. In one such study, Hodgetts et al. (2019) reported that healthy young adult carriers of the apolipoprotein-E (APOE) ε4 allele – the strongest common genetic risk factor for AD – showed higher FA and lower MD in the PHCB but not the inferior longitudinal fasciculus (ILF). These results are consistent with proposals claiming that heightened neural activity and intrinsic connectivity play a significant role in increasing posteromedial cortex vulnerability to amyloid-β and tau spread beyond the medial temporal lobe. Given the implications for understanding AD risk, here we sought to replicate Hodgetts et al.‘s finding in a larger sample (N = 128; 40 APOE ε4 carriers, 88 APOE ε4 non-carriers) of young adults (age range = 19–33). Extending this work, we also conducted an exploratory analysis using a more advanced measure of white matter microstructure: hindrance modulated orientational anisotropy (HMOA). Contrary to the original study, we did not observe higher FA or lower MD in the PHCB of APOE ε4 carriers relative to non-carriers. Bayes factors (BFs) further revealed moderate-to-strong evidence in support of these null findings. In addition, we observed no APOE ε4-related differences in PHCB HMOA. Our findings indicate that young adult APOE ε4 carriers and non-carriers do not differ in PHCB microstructure, casting some doubt on the notion that early-life variation in PHCB tract microstructure might enhance vulnerability to amyloid-β accumulation and/or tau spread.

Invasive tract-tracing studies in rodents implicate a direct connection between the subiculum and bed nucleus of the stria terminalis (BNST) as a key component of neural pathways mediating hippocampal regulation of the Hypothalamic-Pituitary-Adrenal (HPA) axis. A clear characterisation of the connections linking the subiculum and BNST in humans and non-human primates is lacking. To address this, we first delineated the projections from the subiculum to the BNST using anterograde tracers injected into macaque monkeys, revealing evidence for a monosynaptic subiculum-BNST projection involving the fornix. Second, we used in vivo diffusion MRI tractography in macaques and humans to demonstrate substantial subiculum complex connectivity to the BNST in both species. This connection was primarily carried by the fornix, with additional connectivity via the amygdala, consistent with rodent anatomy. Third, utilising the twin-based nature of our human sample, we found that microstructural properties of these tracts were moderately heritable (h2 ∼ 0.5). In a final analysis, we found no evidence of any significant association between subiculum complex-BNST tract microstructure and indices of perceived stress/dispositional negativity and alcohol use, derived from principal component analysis decomposition of self-report data. Our findings address a key translational gap in our knowledge of the neurocircuitry regulating stress.

Abstract Background Professional society guidelines recommend risk-reducing salpingo-oophorectomy (RRSO) for women with pathogenic variants (PVs) in ovarian cancer-risk genes. Personalization of that intervention is based on gene-specific phenotypes; however, the age of ovarian cancer diagnosis in women with PVs in moderate penetrance ovarian cancer-risk genes is not well characterized. Women who had hereditary cancer panel testing from September 2013–May 2019 were included (N = 631,950). Clinical/demographic information was compared for women with a PV in BRIP1, RAD51C, or RAD51D versus in BRCA1 or BRCA2. Results PVs in BRIP1, RAD51C, or RAD51D were identified in 0.5% of all tested women but in 1.6% of women with a history of ovarian cancer (~ 3-fold increase). PVs in BRCA1 or BRCA2 were identified in 2.4% of all tested women but in 6.1% of women with a history of ovarian cancer (~ 2.5-fold increase). The proportion of women with a personal or family history of ovarian cancer was similar among women with a PV in BRIP1, RAD51C, RAD51D, BRCA1, or BRCA2. The median age at ovarian cancer diagnosis was 53 years for BRCA1, 59 years for BRCA2, 65 years for BRIP1, 62 years for RAD51C, and 57 years for RAD51D. Conclusions These data reinforce the importance of identifying PVs in moderate penetrance ovarian cancer-risk genes. The age at ovarian cancer diagnosis was older for women with PVs in BRIP1, RAD51C, or RAD51D, suggesting that it is safe to delay RRSO until age 45–50 in RAD51D PV carriers and possibly until age 50–55 in BRIP and RAD51C PV carriers.

Abstract Copy number variants are amongst the most highly penetrant risk factors for psychopathology and neurodevelopmental deficits, but little information about the detailed clinical phenotype associated with particular variants is available. We present the largest study of the microdeletion and -duplication at the distal 1q21 locus, which has been associated with schizophrenia and intellectual disability, in order to investigate the range of psychiatric phenotypes. Clinical and cognitive data from 68 deletion and 55 duplication carriers were analysed with logistic regression analysis to compare frequencies of mental disorders between carrier groups and controls, and linear mixed models to compare quantitative phenotypes. Both children and adults with copy number variants at 1q21 had high frequencies of psychopathology. In the children, neurodevelopmental disorders were most prominent (56% for deletion, 68% for duplication carriers). Adults had increased prevalence of mood (35% for deletion [OR = 6.6 (95% CI: 1.4–40.1)], 55% for duplication carriers [8.3 (1.4–55.5)]) and anxiety disorders (24% [1.8 (0.4–8.4)] and 55% [10.0 (1.9–71.2)]). The adult group, which included mainly genetically affected parents of probands, had an IQ in the normal range. These results confirm high prevalence of neurodevelopmental disorders associated with CNVs at 1q21 but also reveal high prevalence of mood and anxiety disorders in a high-functioning adult group with these CNVs. Because carriers of neurodevelopmental CNVs who show relevant psychopathology but no major cognitive impairment are not currently routinely receiving clinical genetic services widening of genetic testing in psychiatry may be considered.

Improvements in retail shelf life of exported beef will help with merchandising and increase competitiveness in the worldwide market for United States beef products. The objective of this study was to determine the effect of topically applying acerola cherry powder or rosemary extract from various suppliers on beef bone-in short rib steak and chuck roll steak shelf life. USDA Choice beef bone-in short ribs (IMPS 123A) and chuck rolls (IMPS 116A) were aged (0°C) for 28 d postfabrication. Following aging, 1.02-cm-thick steaks were cut (N = 126) and systematically assigned to a treatment based on steak location within the subprimal. Treatments included untreated control (C), topically sprayed (2 mL) with an acerola cherry powder solution (0.05%) from 1 of 3 suppliers (C1, C2, C3), or topically sprayed (2 mL) with a rosemary extract solution (0.10%) from 1 of 3 suppliers (R1, R2, R3). Half of the steaks were assigned to day 0 lipid oxidation, metmyoglobin-reducing activity (MRA), and oxygen consumption; the remaining steaks were assigned to color evaluation over 4 d of retail display followed by day 4 lipid oxidation and MRA. Short rib steaks treated with antioxidants had a brighter oxygenated lean color than control steaks (P < 0.001). There was an interaction (P = 0.028) between time of retail display and MRA. Short rib steaks treated with C3 and R2 did not change in MRA between day 0 and 4 (P = 0.620, P = 0.428, respectively). Chuck roll steaks treated with C1, C2, C3, R2, and R3 all had a higher, or more desirable, MRA than the control steaks on day 0 (P < 0.001). Applying topical antioxidants improves the shelf-life stability of steaks from beef bone-in short ribs and chuck rolls aged for an extended period.

Variation in cut size and weight of fabricated subprimals is a challenge of increased beef carcass weights. Subsequently, variation in carcass size has resulted in consistency challenges during retail display. The objective of this study was to assess three aging periods of commercially available top rounds from varying carcass weights as it relates to yield and color characteristics. In the current study, 21 industry average weight (AW; 340 to 409 kg; no industry discount) beef carcasses and 21 oversized (OS; exceeding 454 kg; receive a discount) beef carcasses were evaluated. Carcasses were selected at a commercial beef packing plant, where the left and right (paired) top round subprimals of each carcass were procured. Paired top rounds were assigned to a short (8 d), average (23 d), or extended (42 d) postmortem aging period. After wet-aging, subprimals were fabricated into steaks for additional analysis. Steaks were evaluated as whole top round steaks or further fabricated into “ superficial ” and “ deep ” portions at 5.08 cm from the superficial edge of the Semimembranosus and the Adductor muscle. Top rounds and steaks from OS carcasses were larger (P < 0.01) than those from AW carcasses. Quantitative color of the anatomically deep locations of the OS steaks had the greatest mean L* (lightness; P < 0.01), a* (redness; P < 0.01) and b* (yellowness; P < 0.01) values. Extending the aging timeline increased L* (lightness; P < 0.01), decreased a* (redness; P < 0.01), and decreased b* (yellowness; P < 0.01). Alternative top round steak fabrication that separates the deep and superficial anatomical locations could be an effective means of providing more uniform steaks.

Loss of stromal caveolin-1 (Cav-1) is a biomarker of a cancer-associated fibroblast (CAF) phenotype and is related to progression, metastasis, and poor outcomes in several cancers. The objective of this study was to evaluate the clinical significance of Cav-1 expression in invasive epithelial ovarian cancer (OvCa). Epithelial and stromal Cav-1 expression were quantified in serous OvCa and benign ovarian tissue in two, independent cohorts-one quantified expression using immunohistochemistry (IHC) and the other using multiplex immunofluorescence (IF) with digital image analysis designed to target CAF-specific expression. Cav-1 expression was significantly downregulated in OvCa stroma compared to non-neoplastic stroma using both the IHC (p = 0.002) and IF (p = 1.8x10-13) assays. OvCa stroma showed Cav-1 downregulation compared to tumor epithelium with IHC (p = 1.2x10-24). Conversely, Cav-1 expression was higher in OvCa stroma compared to tumor epithelium with IF (p = 0.002). There was moderate correlation between IHC and IF methods for stromal Cav-1 expression (r2 = 0.69, p = 0.006) whereas there was no correlation for epithelial expression (r2 = 0.006, p = 0.98). Irrespective of the staining assay, neither response to therapy or overall survival correlated with the expression level of Cav-1 in the stroma or tumor epithelium. Our findings demonstrate a loss of stromal Cav-1 expression in ovarian serous carcinomas. Studies are needed to replicate these findings and explore therapeutic implications, particularly for immunotherapy response.

The objective of this study was to evaluate the effect of timing of electrical stimulation on free calcium concentration, calpain-2 activity, Warner-Bratzler shear force (WBSF), and consumer sensory analysis. Twenty-three beef steers were harvested and stimulated (S) using extra-low voltage or not stimulated (NS), at exsanguination and/or 1 h postmortem, resulting in 4 different stimulation treatments: NS-NS, NS-S, S-NS, or S-S. Samples were removed from the longissimus lumborum (LL) and semimembranosus (SM) for free calcium and calpain-2 analysis on days 1, 4, and 14 postmortem. WBSF and sensory analysis steaks were removed on day 4 and frozen (4 d) or aged to 14 d postmortem. Data were analyzed using the mixed model or generalized linear mixed model procedure of SAS (SAS Institute, Inc., Cary, NC), with significance determined at P < 0.05. There was a tendency for an aging-period-by-stimulation-treatment interaction for LL free calcium concentration (P = 0.05), and there was a significant difference between aging periods (P < 0.01). No difference was observed in free calcium concentration in the SM between stimulation treatments (P = 0.44); aging, however, significantly increased SM free calcium concentration (P < 0.01). Stimulation did not impact native calpain-2 activity in the LL (P = 0.71) or SM (P = 0.89). Stimulation treatment did not improve tenderness values for WBSF analysis for the LL (P = 0.69) or SM (P = 0.61) or consumer sensory analysis in the LL (P = 0.56) or SM (P = 0.36). A longer aging period tended to increase calpain-2 activity in the SM (P = 0.08), improve WBSF in the LL (P = 0.09), and significantly improve consumer tenderness scores in the SM (P < 0.01). In conclusion, the timing of electrical stimulation utilized in the current study tended to influence free calcium concentration in the LL but did not influence calpain-2 activity or beef tenderness. Aging, however, improved tenderness.

Genetic panel use as a selection tool has grown in popularity in the beef industry. The objective of the study was to determine whether beef cattle genetically selected for tenderness generated a tender product. Igenity® (IT) panel results were provided by a cattle producer for 52 steers, which were harvested at a commercial harvest facility. Boneless strip loins (Institutional Meat Purchase Specifications #180; United States Department of Agriculture [USDA] Choice, n = 32; USDA Prime n = 20) were collected from the left side of each carcass and transported to the University of Idaho Meat Science Laboratory. Four steaks were cut from each subprimal and assigned to aging periods of 7, 14, and 21 d for Warner-Bratzler Shear Force (WBSF) analysis or 21 d for consumer sensory analysis. Carcasses were assigned to tenderness groups based on their IT tenderness indexes (Low IT, 3 – 6, n = 30; High IT, 7 – 10, n = 22). Data were analyzed using the mixed model procedure of SAS version 9.4 (SAS Institute Inc., Cary, NC). An interaction was observed between tenderness group and USDA quality grade (P = 0.015) when analyzing WBSF. All of the cattle had less than 4.14 kg of WBSF; however, USDA Prime steers that were in the High IT tenderness group produced more tender steaks than High IT USDA Choice, Low IT USDA Prime, and Low IT USDA Choice steers. Consumers were not able to detect tenderness differences between IT tenderness groups (P = 0.11) or USDA quality grades (P = 0.11), but they found USDA Prime steaks to be more acceptable (P = 0.01), juicier (P = 0.01), and more flavorful (P = 0.02) than USDA Choice steaks. In conclusion, regardless of tenderness group, USDA Prime steaks were preferred by consumers over USDA Choice steaks in terms of flavor, juiciness, and acceptability.

The essay contains a stylistic analysis of the dispirited black female. Hurston uses Their Eyes Were Watching God to deconstruct the negative image of the dispirited black female, a woman who is dogged by a tragic past. True to her commitment to not fall under the constraints of feeling ―tragically colored,‖ Hurston uses Nanny and an empowered sermon to create a warrior woman who struggles to hold on to the remnants of a spirit that had been beaten down by the effects and after effects of slavery. While there are a plethora of articles on Nanny, there are no articles (at least none that were found after quite an exhaustive search) that focus on a stylistic study of Nanny‘s sermon. In an effort to add to the scholarship for Nanny, this paper analyses Nanny‘s sermon—the independent and dependent clauses, the signifiers, and the cohesive ties—all of which help Nanny shed the burdens of her past, whereby freeing her from the burdens of the dispirited black woman.

We studied resting-state oscillatory connectivity using magnetoencephalography in healthy young humans (N = 183) genotyped for APOE-ɛ4, the greatest genetic risk for Alzheimer’s disease (AD). Connectivity across frequencies, but most prevalent in alpha/beta, was increased in APOE-ɛ4 in a set of mostly right-hemisphere connections, including lateral parietal and precuneus regions of the Default Mode Network. Similar regions also demonstrated hyperactivity, but only in gamma (40–160 Hz). In a separate study of AD patients, hypoconnectivity was seen in an extended bilateral network that partially overlapped with the hyperconnected regions seen in young APOE-ɛ4 carriers. Using machine-learning, AD patients could be distinguished from elderly controls with reasonable sensitivity and specificity, while young APOE-e4 carriers could also be distinguished from their controls with above chance performance. These results support theories of initial hyperconnectivity driving eventual profound disconnection in AD and suggest that this is present decades before the onset of AD symptomology.