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The continuing emergence of variants of the SARS-CoV-2 virus requires the development of modular molecular therapies. Here, we engineered a recombinant amphiphilic protein, oleosin, to spontaneously self-assemble into multivalent micellar nanostructures which can block the Spike S1 protein of SARS-CoV-2 pseudoviruses (PVs). Short recombinant proteins like oleosin can be formulated more easily than antibodies and can be functionalized with precision through genetic engineering. We cloned S1-binding mini-protein genes called LCB

The incorporation of light-responsive domains into engineered proteins has enabled control of protein localization, interactions, and function with light. We integrated optogenetic control into proximity labeling (PL), a cornerstone technique for high-resolution proteomic mapping of organelles and interactomes in living cells. Through structure-guided screening and directed evolution, we installed the light-sensitive LOV domain into the PL enzyme TurboID to rapidly and reversibly control its labeling activity with low-power blue light. “LOV-Turbo” works in multiple contexts and dramatically reduces background in biotin-rich environments such as neurons. We used LOV-Turbo for pulse-chase labeling to discover proteins that traffick between endoplasmic reticulum, nuclear, and mitochondrial compartments under cellular stress. We also showed that instead of external light, LOV-Turbo can be activated by BRET from luciferase, enabling interaction-dependent PL. Overall, LOV-Turbo increases the spatial and temporal precision of PL, expanding the scope of experimental questions that can be addressed with PL.

Supplementary Tables 1-3, Supplementary Figures 1 and 2, Supplementary Data 1 and 2

Endocrine therapy is underutilized to reduce breast cancer incidence among women at increased risk. Polygenic risk scores (PRSs) assessing 77 breast cancer genetic susceptibility loci personalizes risk estimates. We examined effect of personalized PRS breast cancer risk prediction on intention to take and endocrine therapy uptake among women at increased risk. Eligible participants had a 10-year breast cancer risk ≥5% by Tyrer–Cuzick model [International Breast Cancer Intervention Study (IBIS)] or ≥3.0 % 5-year Gail Model risk with no breast cancer history or hereditary breast cancer syndrome. Breast cancer risk was estimated, endocrine therapy options were discussed, and endocrine therapy intent was assessed at baseline. After genotyping, PRS-updated breast cancer risk estimates, endocrine therapy options, and intent to take endocrine therapy were reassessed; endocrine therapy uptake was assessed during follow-up. From March 2016 to October 2017, 151 patients were enrolled [median (range) age, 56.1 (36.0–76.4 years)]. Median 10-year and lifetime IBIS risks were 7.9% and 25.3%. Inclusion of PRS increased lifetime IBIS breast cancer risk estimates for 81 patients (53.6%) and reduced risk for 70 (46.4%). Of participants with increased breast cancer risk by PRS, 39 (41.9%) had greater intent to take endocrine therapy; of those with decreased breast cancer risk by PRS, 28 (46.7%) had less intent to take endocrine therapy (P < 0.001). On multivariable regression, increased breast cancer risk by PRS was associated with greater intent to take endocrine therapy (P < 0.001). Endocrine therapy uptake was greater among participants with increased breast cancer risk by PRS (53.4%) than with decreased risk (20.9%; P < 0.001). PRS testing influenced intent to take and endocrine therapy uptake. Assessing PRS effect on endocrine therapy adherence is needed.Prevention Relevance: Counseling women at increased breast cancer risk using polygenic risk score (PRS) risk estimates can significantly impact preventive endocrine therapy uptake. Further development of PRS testing to personalize breast cancer risk assessments and endocrine therapy counselling may serve to potentially reduce the incidence of breast cancer in the future.

Supplementary Figure 3 from Agglomerative Epigenetic Aberrations Are a Common Event in Human Breast Cancer

Supplementary Figure 2 from Agglomerative Epigenetic Aberrations Are a Common Event in Human Breast Cancer

Supplementary Table 1 from Agglomerative Epigenetic Aberrations Are a Common Event in Human Breast Cancer

Supplementary Figure 1 from Agglomerative Epigenetic Aberrations Are a Common Event in Human Breast Cancer

Coordinates of All Identified DRMs from Agglomerative Epigenetic Aberrations Are a Common Event in Human Breast Cancer

The experience of pursuing a graduate seminary education is rife with many stressors, including the heavy financial burden required to fund a seminary degree. Shame, understood as an experience of being unworthy and inadequate at one’s core, may be a natural reaction to financial hardship as many individuals may believe their inadequacy is the cause of their financial difficulties. The present study assessed 189 graduate seminary students from institutions accredited by the Association of Theological Schools to further understand the relationships between shame, financial distress, depression, anxiety, and spiritual well-being. Based on previous research, we hypothesized that financial well-being and shame would be negatively correlated. Furthermore, we hypothesized that shame would mediate the relationship between financial well-being and a number of negative outcomes, including depression, anxiety, and lack of spiritual well-being. Our findings demonstrated that the seminary population experienced shame in regard to perceived lack of financial well-being; shame was significantly and negatively related to financial well-being and significantly mediated the relationship between financial well-being and depression, anxiety, and spiritual well-being. These results indicate that financial hardship, including student debt, appears to have a far-reaching impact that causes distress on various levels for seminary students and that shame may be an important mechanism in understanding how this happens.

This study phenomenologically explored 36 embryo adoption couples’ experience of choosing a donor and considering what kind of connection, if any, they wished to have with the donor. Thirty-six emb...

Mentalising has long been suggested to play an important role in irony interpretation. We hypothesised that another important cognitive underpinning of irony interpretation is likely to be children's capacity for mental set switching – the ability to switch flexibly between different approaches to the same task. We experimentally manipulated mentalising and set switching to investigate their effects on the ability of 7-year-olds to determine if an utterance is intended ironically or literally. The component of mentalising examined was whether the speaker and listener shared requisite knowledge.We developed a paradigm in which children had to select how a listener might reply, depending on whether the listener shared knowledge needed to interpret the utterance as ironic. Our manipulation of requisite set switching found null results. However, we are the first to show experimentally that children as young as seven years use mentalising to determine whether an utterance is intended ironically or literally.

Loss of sensory hair cells causes permanent hearing and balance deficits in humans and other mammals, but for nonmammals such deficits are temporary. Nonmammals recover hearing and balance sensitivity after supporting cells proliferate and differentiate into replacement hair cells. Evidence of mechanical differences between those sensory epithelia and their supporting cells prompted us to investigate whether the capacity to activate YAP, an effector in the mechanosensitive Hippo pathway, correlates with regenerative capacity in acceleration-sensing utricles of chickens and mice of both sexes. After hair cell ablation, YAP accumulated in supporting cell nuclei in chicken utricles and promoted regenerative proliferation, but YAP remained cytoplasmic and little proliferation occurred in mouse utricles. YAP localization in supporting cells was also more sensitive to shape change and inhibition of MST1/2 in chicken utricles than in mouse utricles. Genetic manipulations showed thatin vivoexpression of the YAP-S127A variant caused robust proliferation of neonatal mouse supporting cells, which produced progeny that expressed hair cell markers, but proliferative responses declined postnatally. Expression of YAP-5SA, which more effectively evades inhibitory phosphorylation, resulted in TEAD-dependent proliferation of striolar supporting cells, even in adult utricles. Conditional deletion of LATS1/2 kinases abolished the inhibitory phosphorylation of endogenous YAP and led to striolar proliferation in adult mouse utricles. The findings suggest that damage overcomes inhibitory Hippo signaling and facilitates regenerative proliferation in nonmammalian utricles, whereas constitutive LATS1/2 kinase activity suppresses YAP-TEAD signaling in mammalian utricles and contributes to maintaining the proliferative quiescence that appears to underlie the permanence of sensory deficits.SIGNIFICANCE STATEMENTLoud sounds, ototoxic drugs, infections, and aging kill sensory hair cells in the ear, causing irreversible hearing loss and balance deficits for millions. In nonmammals, damage evokes shape changes in supporting cells, which can divide and regenerate hair cells. Such shape changes are limited in mammalian ears, where supporting cells develop E-cadherin-rich apical junctions reinforced by robust F-actin bands, and the cells fail to divide. Here, we find that damage readily activates YAP in supporting cells within balance epithelia of chickens, but not mice. Deleting LATS kinases or expressing YAP variants that evade LATS-mediated inhibitory phosphorylation induces proliferation in supporting cells of adult mice. YAP signaling eventually may be harnessed to overcome proliferative quiescence that limits regeneration in mammalian ears.

Manitoba implemented the first Canadian provincial program of reflex screening through mismatch repair immunohistochemistry (MMR-IHC) for all colorectal cancers diagnosed at age 70 years or younger in December 2017. We evaluated compliance to universal reflex testing and for referrals to Genetics for individuals with MMR-deficient tumors.We searched the provincial pathology database with "adenocarcinoma" in the colorectal specimen pathology reports between March 2018 and December 2020. We cross-referenced with paper and electronic records in the Program of Genetics and Metabolism to determine whether patients with MMR-deficient tumors had been referred for Genetic assessment and what proportion of patients and first-degree relatives accepted an appointment and genetic testing. We performed logistic regression analysis to identify predictors of testing.We identified 3,146 colorectal adenocarcinoma specimens (biopsies and surgical resections) from 1,692 unique individuals (mean age 68.66 years, male 57%). Of those aged 70 years or younger (n = 936), 89.4% received MMR-IHC screening. Individual pathologists (categorized by the highest, average, and lowest screening rates) were the biggest predictors of MMR-IHC screening on multivariable analysis (highest vs lowest: odds ratio 17.5, 95% confidence interval 6.05-50.67). While only 53.4% (n = 31) of 58 screen-positive cases were referred by pathologists for genetic assessment, other clinicians referred an additional 22.4% (n = 13), resulting in 75.8% overall referral rate of screen-positive cases. Thirteen (1.4%) patients (1.1%, aged 70 years or younger) were confirmed to experience Lynch syndrome through germline testing, and 8 first-degree relatives (an average of 1.6 per patient) underwent cascade genetic testing.The first Canadian Lynch syndrome screening program has achieved high rates of reflex testing.

Advanced pancreatic cancer (APC) disproportionately impacts older adults. Randomized trials demonstrate improved overall survival (OS) with combination chemotherapy including 5-fluorouracil, irinotecan, leucovorin, and oxaliplatin (FOLFIRINOX) or nab-paclitaxel and gemcitabine compared with gemcitabine alone, but with increased toxicity. Older adults are at increased risk of side effects from chemotherapy. The aim of this study was to assess the efficacy and toxicity of chemotherapy in older adults with APC.Patients diagnosed with APC from 2011 to 2016 were identified using the Manitoba Cancer Registry. Patient and treatment characteristics, toxicity, and outcomes of patients 65 years of age and above treated with palliative chemotherapy were compared by treatment regimen. OS was assessed using the Kaplan-Meier method. A Cox regression was used to identify independent predictors of OS.A total of 87 patients aged 65 years and above received palliative chemotherapy: 52 (59.7%) FOLFIRINOX, 21 (24.1%) nab-paclitaxel and gemcitabine, and 14 (16.1%) gemcitabine, with a median age of 69 (65 to 84), 75 (65 to 88), and 73 (67 to 82), Eastern Cooperative Oncology Group (ECOG) performance status difference in hematologic toxicity between regimens (P=0.807). An increase in nonhematologic toxicity was seen with FOLFIRINOX (P0.001), specifically neuropathy (P=0.008), fatigue (P0.001), and nausea/vomiting (P=0.008). FOLFIRINOX was associated with improved radiologic response (P=0.05) and OS (P=0.035). PS, baseline carbohydrate antigen 19-9 level, and chemotherapy regimen were independent predictors of survival.FOLFIRINOX is associated with improved response and OS in older adults with APC. FOLFIRINOX has a manageable safety profile in this population and should be considered in fit older adults with APC.

The loss of inner ear hair cells causes permanent hearing and balance deficits in humans and other mammals, but non-mammals recover after supporting cells (SCs) divide and replace hair cells. The proliferative capacity of mammalian SCs declines as exceptionally thick circumferential F-actin bands develop at their adherens junctions. We hypothesized that the reinforced junctions were limiting regenerative responses of mammalian SCs by impeding changes in cell shape and epithelial tension. Using micropipette aspiration and atomic force microscopy, we measured mechanical properties of utricles from mice and chickens. Our data show that the epithelial surface of the mouse utricle stiffens significantly during postnatal maturation. This stiffening correlates with and is dependent on the postnatal accumulation of F-actin and the cross-linker Alpha-Actinin-4 at SC-SC junctions. In chicken utricles, where SCs lack junctional reinforcement, the epithelial surface remains compliant. There, SCs undergo oriented cell divisions and their apical surfaces progressively elongate throughout development, consistent with anisotropic intraepithelial tension. In chicken utricles, inhibition of actomyosin contractility led to drastic SC shape change and epithelial buckling, but neither occurred in mouse utricles. These findings suggest that species differences in the capacity for hair cell regeneration may be attributable in part to the differences in the stiffness and contractility of the actin cytoskeletal elements that reinforce adherens junctions and participate in regulation of the cell cycle.

Mental labor is an under-researched and long-invisible component of family work. Scholars have described mental labor as important, taxing, and disproportionately performed by mothers compared to fathers. However, operational definitions used in these studies were only preliminary and lack unified terminology. Answering calls for expanded views of household labor and better definitions of its content, we undertook a phenomenological investigation of family-related mental labor performed by mothers. Our interpretive phenomenological analysis of seven focus group interviews produced a definition of mothers’ mental labor and its component parts that was empirically grounded in the lived experience of mothers of young children ( N = 25). Distinct from housework chores, childcare, and emotion work, mental labor emerged as thinking activity performed for the sake of accomplishing family goals. We identified six forms of mental labor: (a) planning and strategizing, (b) monitoring and anticipating needs, (c) metaparenting, (d) knowing (learning and remembering), (e) managerial thinking (including delegating and instructing), and (f) self-regulation. All participating mothers represented themselves as their family’s primary mental laborer, regardless of employment status or their partner’s level of involvement. Our description of mental labor may help parents, researchers, clinicians, policy makers, and educators recognize, value, and better account for the mental labor dynamics operating in the construction of family life, reproduction of gender roles, and perpetuation of gender gaps in family labor division and mental load. Online slides for instructors who want to use this article for teaching are available on PWQ's website at http://journals.sagepub.com/page/pwq/suppl/index

Necrotizing enterocolitis is a major cause of mortality and morbidity in the preterm infant population. The gut microbiome is of particular interest in research surrounding necrotizing enterocolitis, because variations in the intestinal microbiota seem to correlate with the risk of inflammation and disease. Recent advances in non-culture-based genomic sequencing have also allowed for more intricate analyses of the intestinal microbiome. Its evolution seems to be influenced by intrauterine and extrauterine factors, ranging from antenatal antibiotic exposure to type of enteral feeds. Ultimately, these alterations in the gut microbiome have the potential to result in devastating diseases like necrotizing enterocolitis.

Designed protein receptors hold diagnostic and therapeutic promise. We now report the design of five consensus leucine-rich repeat proteins sCLRR4-8) based on the LRR domain of nucleotide-binding oligomerization domain (NOD)-like receptors involved in the innate immune system. The CLRRs bind muramyl dipeptide (MDP), a bacterial cell wall component, with micromolar affinity. The overall Kd app values ranged from 1.0 to 57 μM as measured by fluorescence quenching experiments. Biphasic fluorescence quenching curves were observed in all CLRRs with higher affinity Kd1 values ranging from 0.04 to 4.5 μM, and lower affinity Kd2 values ranging from 3 to 227 μM. These biphasic binding curves, along with the docking studies of MDP binding to CLRR4, suggest that at least two MDPs bind to each protein. Previously, only single MDP binding was reported. This high-capacity binding of MDP promises small, soluble, stable CLRR scaffolds as candidates for the future design of pathogen biosensors. This article is protected by copyright. All rights reserved.

This study extends the logic of prior studies showing phonetic convergence between interlocutors to the structural domain. We ask whether listeners' adaptation of the syntactic forms they produce depends on their perceptions about their interlocutor's social proximity and linguistic competence, using structural priming as a measure of convergence. Two experiments compared structural priming in dialogues between native British English speakers and (i) other native British English speakers, (ii) native speakers of North American English, and (iii) non-native speakers of English, to assess to what extent interlocutor characteristics influence structural convergence in dialogue. Our findings suggest that rates of structural convergence depend both on a speaker's pre-existing structural biases for particular verbs, and their perception of (linguistic or social) similarity to their interlocutor. This suggests that low-level mechanisms underlying structural convergence may be mediated by beliefs about how interlocutors are socially situated with respect to each other. [ABSTRACT FROM AUTHOR] Copyright of Language, Cognition & Neuroscience is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This randomized clinical trial investigates whether withholding the standard 48 hours of intravenous empirical antibiotics immediately after birth in preterm infants protects the developing microbiome and improves clinical outcomes.

SUMMARYThe ability to record transient cellular events in the DNA or RNA of cells would enable precise, large-scale analysis, selection, and reprogramming of heterogeneous cell populations. Here we report a molecular technology for stable genetic tagging of cells that exhibit activity-related increases in intracellular calcium concentration (FLiCRE). We used FLiCRE to transcriptionally label activated neural ensembles in the nucleus accumbens of the mouse brain during brief stimulation of aversive inputs. Using single-cell RNA sequencing, we detected FLiCRE transcripts among the endogenous transcriptome, providing simultaneous readout of both cell-type and calcium activation history. We identified a cell-type in the nucleus accumbens activated downstream of long-range excitatory projections. Taking advantage of FLiCRE’s modular design, we expressed an optogenetic channel selectively in this cell-type, and showed that direct recruitment of this otherwise genetically-inaccessible population elicits behavioral aversion. The specificity and minute-resolution of FLiCRE enables molecularly-informed characterization, manipulation, and reprogramming of activated cellular ensembles.

Limited knowledge is available on the relationship between antigen-specific immune responses and COVID-19 disease severity. We completed a combined examination of all three branches of adaptive immunity at the level of SARS-CoV-2-specific CD4+ and CD8+ T cell and neutralizing antibody responses in acute and convalescent subjects. SARS-CoV-2-specific CD4+ and CD8+ T cells were each associated with milder disease. Coordinated SARS-CoV-2-specific adaptive immune responses were associated with milder disease, suggesting roles for both CD4+ and CD8+ T cells in protective immunity in COVID-19. Notably, coordination of SARS-CoV-2 antigen-specific responses was disrupted in individuals > 65 years old. Scarcity of naive T cells was also associated with ageing and poor disease outcomes. A parsimonious explanation is that coordinated CD4+ T cell, CD8+ T cell, and antibody responses are protective, but uncoordinated responses frequently fail to control disease, with a connection between ageing and impaired adaptive immune responses to SARS-CoV-2., Highlights - Adaptive immune responses limit COVID-19 disease severity - Multiple coordinated arms of adaptive immunity control better than partial responses - CXCL10 may be a biomarker of impaired T cell responses in acute COVID-19 - Aging and scarcity of naive T cells may be linked risk factors for severe COVID-19, Analysis of SARS-CoV-2 specific adaptive immune responses during acute COVID-19 identifies coordination between SARS-CoV-2-specific CD4 T cells and CD8 T cells to limit disease severity. Aged individuals often exhibit uncoordinated adaptive responses, potentially tied to scarcity of naive T cells, highlighting immunologic risk factors linked to disease severity.

The ability to record transient cellular events in the DNA or RNA of cells would enable precise, large-scale analysis, selection, and reprogramming of heterogeneous cell populations. Here, we report a molecular technology for stable genetic tagging of cells that exhibit activity-related increases in intracellular calcium concentration (FLiCRE). We used FLiCRE to transcriptionally label activated neural ensembles in the nucleus accumbens of the mouse brain during brief stimulation of aversive inputs. Using single-cell RNA sequencing, we detected FLiCRE transcripts among the endogenous transcriptome, providing simultaneous readout of both cell-type and calcium activation history. We identified a cell type in the nucleus accumbens activated downstream of long-range excitatory projections. Taking advantage of FLiCRE's modular design, we expressed an optogenetic channel selectively in this cell type and showed that direct recruitment of this otherwise genetically inaccessible population elicits behavioral aversion. The specificity and minute resolution of FLiCRE enables molecularly informed characterization, manipulation, and reprogramming of activated cellular ensembles.

Background: The objective of this study was to quantify the influence of race/ethnicity and socioeconomic status (SES) on overall mortality within an insured population of patients diagnosed with melanoma in Southern California. Specifically, we compared outcomes between insured patients within the largest vertically integrated health care system in Southern California, Kaiser Permanente Southern California (KPSC), with insured patients diagnosed elsewhere. Methods: We identified adults (≥20 years old) diagnosed with Stage 0-IV melanoma from 1 January 2009 through 31 December 2014, followed through the end of 2017, from the California Cancer Registry. We compared overall mortality in those diagnosed within KPSC versus those with other private insurance (OPI). Main variables included race/ethnicity and SES based on 2010 Census data. Person-year (PY) mortality rates and 95% confidence intervals (CI) were calculated by race/ethnicity and SES. Multivariable-adjusted hazard ratios (HR) for the association between all-cause mortality and race/ethnicity and SES were estimated using Cox proportional hazards models. Results: A total of 14,614 adults were diagnosed with melanoma in our cohort (KPSC: n=4,701; OPI: n=9,913). The total of number of all-cause deaths was 2,456 (KPSC: n=729; OPI: n=1,727). Mortality rates by race/ethnicity within KPSC and OPI did not reveal significant disparities. The poorest patients in KPSC (lowest, lower-middle, and middle SES groups) had statistically significant decreased mortality rates compared to those in OPI. For example, in the lowest SES, those in OPI had 38.7 additional deaths per 1000 PY compared to KPSC (KPSC Rate per 1000 PY: 57.7, 95% CI: 44.9-73.1; OPI Rate per 1000 PY: 96.4, 95% CI 80.9-113.9). Multivariable models demonstrated statistically significant increased mortality risk among the lowest (HR 1.70, 95% CI 1.43-2.02), lower-middle (HR 1.47, 9% CI 1.29-1.68), middle (HR 1.36, 95% CI 1.21-1.53), and upper-middle (HR 1.19, 95% CI 1.07-1.33) SES groups when compared to the highest SES group. This increased mortality risk among lower SES groups persisted when stratifying by KPSC and OPI. Although KPSC patients had a lower mortality risk across all SES groups compared to OPI, the difference was not statistically significant. For example, among the lowest SES, those in the OPI group had an overall mortality risk 80% greater than those of the highest SES (HR 1.80; 95% CI 1.47-2.22), while those in KPSC held risk only 47% greater (HR 1.47, 95% CI 1.09-2.00). Conclusions: While the existence of disparities in cancer survival are well established, our present study examined survival with respect to SES among two distinct populations of melanoma patients: insured patients within an integrated healthcare system and insured patients within a traditional model of healthcare. While racial/ethnic disparities are not observed in integrated healthcare systems such as KPSC, our findings underscore the persistence of socioeconomic disparities within an insured population, despite having access to care. Citation Format: Amanda H. Rosenthal, Shivani Reddy, Christina N. Kim, Robert M. Cooper, Reina Haque. Healthcare disparities in melanoma overall survival evaluated by race/ethnicity and socioeconomic status and the impact of integrated healthcare on an insured population [abstract]. In: Proceedings of the AACR Virtual Conference: 14th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2021 Oct 6-8. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PO-210.