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Municipal wastewater effluent is one of the largest sources of pollution entering surface waters in the Laurentian Great Lakes. Exposure to wastewater effluent has been associated with impaired immune systems and induction of genotoxicity to aquatic animals. Due to habitat degradation and environmental pollution linked to industrial development and population growth, several regions of the Great Lakes have been designated Areas of Concern (AOCs). In this study, we assessed the effect of extracts of sewage influent, (treated) effluent and receiving surface waters from the Hamilton Harbour AOC and the Toronto and Region AOC (Ontario, Canada) on the phagocytic immune response of rainbow trout (Oncorhynchus mykiss) kidney leukocytes and the genotoxicity (DNA strand breaks) of these extracts on freshwater mussel (Eurynia dilatata) hemocytes. We identified and quantified numerous chemicals present in the various samples extracted for exposure. In freshwater mussels, extracts from Hamilton Harbour AOC induced DNA damage with the most frequency (12 out of 28 samples) regardless of sample type, reflecting past and present industrial activities. In contrast, extracts from Toronto and Region AOC induced DNA damage infrequently (2 out of 32 (summer) and 5 out of 32 (fall) samples, respectively) and from different WWTPs at different times. None of the extracts induced any significant effect on phagocytosis of rainbow trout kidney leukocytes. The present study indicates that despite overall improvements to effluent quality, treatment of influent by WWTPs may not result in a corresponding improvement of the genotoxicity of effluents. In vitro bioassays are useful and cost-effective rapid-screening tools for preliminary assessments of contamination of aquatic ecosystems.

Columbid alphaherpesvirus 1 (CoHV1) is associated with oral or upper respiratory tract lesions, encephalitis, and occasional fatal systemic disease in naive or immunosuppressed pigeons. Clinical disease is often reported with CoHV1 and coinfecting viruses, including pigeon circovirus (PiCV), which may cause host immunosuppression and augment lesion development. A natural outbreak of CoHV1 and PiCV coinfection occurred in a flock of 60 racing rock pigeons ( Columba livia), in which 4 pigeons succumbed within 7 d of clinical onset. Lesions included suppurative stomatitis, pharyngitis, cloacitis, meningitis, and tympanitis, with eosinophilic intranuclear inclusion bodies consistent with herpesviral infection. In addition, large numbers of botryoid intracytoplasmic inclusion bodies were present in the skin, oral mucosa, and bursa of Fabricius, suggestive of circoviral infection, which was confirmed by immunohistochemistry. The concurrent viral load of CoHV1 and PiCV was high in liver, oropharynx, and bursa of Fabricius. We found PiCV in oro-cloacal swabs from 44 of 46 additional birds of variable clinical status, PiCV alone in 23 birds, and coinfection with CoHV1 in 21 birds. Viral copy numbers were significantly higher ( p

Context In ecosystems where rainfall is episodic or highly seasonal, plant recruitment from a soil-stored seed bank occurs during periods of elevated soil moisture conducive to germination and seedling establishment. The release of seed dormancy in response to environmental conditions has significant consequences for the temperature window over which germination occurs, and as such the timing of germination can vary between years. Aims We aimed to understand in seeds of two species of Rutaceae, Diplolaena dampieri and Rhadinothamnus anceps, how dormancy loss and germination timing is influenced by warm stratification. Methods We tested the germination response to temperatures between 5 and 30°C following increasing durations (1–12 and 4–12 weeks) of warm stratification at 20, 25, or 30°C. Key results Warm stratification for 1–8 weeks at 30°C progressively alleviated seed dormancy in D. dampieri and R. anceps, increasing germination proportion from ~0.1 to ~0.5 in both species. Stratification duration was optimal at 30°C for between 4 and 8 weeks depending on species. Warm stratification was not affected by water stress down to −0.8 Mpa. Application of aerosol smoke did not significantly improve germination, and heat treatments had a negative effect on final germination proportion. Conclusions As dormancy was progressively alleviated, the range of temperatures that support germination increased for D. dampieri and decreased for R. anceps, allowing for confirmation of type 1 and type 2 non-deep physiological dormancy (PD), respectively. Implications Arising from this conclusion, we suggest that in Mediterranean climates, type 1 and 2 PD dictate risk-taking and risk-avoiding ecological strategies by shifting the thermal requirements for germination towards that characteristic of the early- or mid-germination season. Classification of non-deep PD may offer a structured approach to predict how temperature requirements shift during dormancy loss, which will provide insight into seed germination response to year-to-year variation in seasonal environmental conditions.

Introduction TNBC is overrepresented in Black women, and Black patients with TNBC have worse clinical outcomes compared to non-Black patients. This disparity likely results from racial differences in clinical, biological, and demographic features of TNBC and social determinants of health. Neoadjuvant chemoimmunotherapy is current standard of care for high-risk TNBC. However, Black patients have been poorly represented in immunotherapy TNBC trials, making it difficult to assess comparative efficacy of immunotherapy in Black patients. Methods We utilized two TNBC neoadjuvant trials to assess racial differences in the tumor immune microenvironment composition and evaluate impact of race on response to chemotherapy vs chemoimmunotherapy. NeoSTOP trial (NCT02413320) randomized 100 stage I-III TNBC patients to receive neoadjuvant carboplatin/paclitaxel + doxorubicin/cyclophosphamide (CbP+AC) or carboplatin/docetaxel (CbD). NeoPACT trial (NCT03639948) enrolled 120 patients with stage I-III TNBC who received neoadjuvant CbD + pembrolizumab (CbD+P). sTILs were centrally quantified, and RNA extracted from pretreatment tissue was subjected to next-generation sequencing. Relative leukocyte fractions were computed by CIBERSORTx. Factors were tested as predictors of pathologic complete response (pCR) using logistic regression analysis. Event-free survival (EFS) was estimated by the Kaplan-Meier method and compared between groups by log-rank test, followed by Cox regression analysis. Results The study population includes 197 patients with known race, sTILs, and gene expression data (84 patients from NeoSTOP, 113 from NeoPACT). 15/84 (18%) patients in NeoSTOP and 20/113 (18%) patients in NeoPACT self-reported Black race. There was no significant difference in age, T or N stage, or germline BRCA1/2 mutation status by race in either study. Black patients had significantly higher sTILs than non-Black patients (median 40% vs 15%, P=0.048) and were more likely to have ≥20% sTILs than non-Black patients (66% vs 44%, P=0.026). There was no significant difference in pCR by race in NeoSTOP (OR=0.60, 95% CI 0.19-1.84, P=0.37; pCR 47% for Black vs 59% for non-Black). In contrast, in NeoPACT, Black patients had a significantly higher pCR compared to non-Black patients (OR=3.27, 95% CI 1.01-10.64, P=0.049; pCR 79% for Black vs 53% for non-Black). In NeoSTOP, EFS was similar for Black and non-Black patients (3-year EFS 92% and 94%, respectively, HR=0.88, 95% CI 0.11-7.28, P=0.90). In NeoPACT, EFS was numerically higher in Black vs non-Black patients (3-year EFS 93% and 81%, respectively, HR=0.43, 95% CI 0.05-3.36, P=0.40); NeoPACT survival follow-up is ongoing at the time of this report. On CIBERSORTx analysis, Black patients had relative depletion of immunosuppressive pro-tumorigenic M2 macrophages (P=0.005) and CD4+ memory resting T cells (P=0.021) compared to non-Black patients. Conclusions Compared to non-Black patients, Black patients with TNBC are more likely to have immune-enriched tumors with lower relative abundance of immunosuppressive leukocytes. These findings suggest potential for higher relative magnitude of benefit from checkpoint inhibitor therapy in Black compared to non-Black patients. Supporting this biological hypothesis, we noted that Black and non-Black patients had equivalent rates of pCR with neoadjuvant chemotherapy; however, pCR rate among Black patients was significantly higher than in non-Black patients when treated with neoadjuvant chemoimmunotherapy. These findings should be confirmed in other studies and can optimize utilization of neoadjuvant chemoimmunotherapy. Our findings also underscore the importance of efforts to address disparity in access and use of immunotherapy in Black patients. Citation Format: Shane R. Stecklein, Rachel Yoder, Roberto Salgado, Joshua M. Staley, Anne O’Dea, Lauren Nye, Manana Elia, Deepti Satelli, Gregory Crane, Richard McKittrick, Andrew K. Godwin, Qamar Khan, Priyanka Sharma. Black patients with triple negative breast cancer (TNBC) have enriched stromal tumor infiltrating lymphocytes (sTILs) and receive preferential benefit from neoadjuvant immunotherapy [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD1-06.

Introduction: The TNBC-DX risk score includes the 14-gene immunoglobulin (IGG) immune signature, tumor size, and nodal status and has shown prognostic value for survival in early-stage TNBC (B. Conte et al., ESMO Breast 2021). However, currently unknown are the value of the TNBC-DX risk score and IGG immune signature in 1) predicting pathologic complete response (pCR) following neoadjuvant therapy, and 2) predicting outcomes the context of neoadjuvant anti-PD1 treatment. Here, we assessed the IGG signature and the TNBC-DX risk score in patients with TNBC treated with neoadjuvant chemoimmunotherapy (NeoPACT; NCT03639948) and neoadjuvant chemotherapy without immunotherapy (NeoSTOP; NCT02413320). Methods: NeoPACT trial enrolled 120 patients with stage I-III TNBC who received carboplatin (AUC 6) + docetaxel (75 mg/m2) + pembrolizumab (200 mg) every 21 days x 6 cycles. NeoSTOP randomized 100 patients with stage I-III TNBC to two chemotherapy regimens; Arm B of NeoSTOP was included in this correlative study as the chemotherapy regimen was identical to NeoPACT. RNA isolated from pretreatment tumor tissue was subjected to next-generation sequencing. The 14-gene IGG immune signature and TNBC-DX risk score were calculated in silico as previously described. Evaluation of stromal tumor-infiltrating lymphocytes (sTILs) was performed as previously described. Markers were tested for prediction of pCR. Logistic regression analysis was used to examine the effect of multiple variables. Event-free survival (EFS) curves were assessed by the Kaplan-Meier method and groups compared by the log-rank test, followed by Cox regression analysis. Results: In this analysis, 112 patients were treated with chemoimmunotherapy on NeoPACT (node-positive = 38%, pCR rate = 58%). In the NeoPACT trial, the 14-gene IGG signature (as a continuous variable) was significantly associated with improved pCR (odds ratio [OR]=1.105, 95% CI 1.019-1.197, P=0.015 for every 0.2 increment). The pCR rates in IGG-high (≥ median) and IGG-low (< median) groups were 71% and 44%, respectively (OR=3.152, 95% CI 1.420-6.996, P=0.005). In terms of EFS, the 14-gene IGG signature was not prognostic (hazard ratio [HR]=0.507, 95% CI 0.148-1.735, p=0.269). In contrast, TNBC-DX risk score was strongly associated with EFS (HR=5.684, 95% CI 1.226-26.356, P=0.012), even when adjusted for sTILs and pCR status (HR=8.415, 95% CI 1.054-67.169, P=0.044). Estimated 3-year EFS rates in TNBC-DX high and low risk groups (above and below median) were 77% and 89%, respectively (P=0.012). In 43 NeoSTOP patients treated with neoadjuvant chemotherapy only (node-positive = 33%, pCR rate = 53%), no association of IGG signature with pCR or TNBC-DX score with EFS was observed. Finally, we observed a moderate correlation between IGG signature and sTILs in both trial datasets combined (r=0.642, P< 0.001). Conclusions: High expression of the 14-gene IGG immune signature in baseline pretreatment tumor samples in early-stage TNBC is significantly associated with pCR following pembrolizumab-based neoadjuvant chemotherapy. The combination of this signature with tumor burden as assessed by TNBC-DX is prognostic for long-term outcomes. Availability of biomarkers that can predict both pathological response and survival with chemoimmunotherapy can optimize this therapy, and evaluation of this biomarker in larger studies is warranted. Citation Format: Priyanka Sharma, Shane R. Stecklein, Rachel Yoder, Joshua M. Staley, Roberto Salgado, Laia Paré, Benedetta Conte, Fara Brasó-Maristany, Anne O’Dea, Lauren Nye, Manana Elia, Deepti Satelli, Gregory Crane, Richard McKittrick, Qamar Khan, Andrew K. Godwin, Aleix Prat. PD11-07 Association of TNBC-DX scores with outcomes in triple-negative breast cancer (TNBC) treated with neoadjuvant pembrolizumab and chemotherapy: a correlative analysis from NeoPACT and NeoSTOP trials [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD11-07.

Little has been published about the process of decision-making between prosthetists and people with lower limb amputation (LLA). The purpose of this study is to identify decisions and factors influencing decision-making in prosthetic rehabilitation from the perspectives of prosthetists and prosthesis users, to identify barriers and opportunities for shared decision-making (SDM).Qualitative semi-structured individual interviews were conducted with 13 prosthetists and 14 prosthesis users from three clinics in three states of the Rocky Mountain and Southwest regions of the United States. Transcripts were analyzed using thematic analysis.Four main themes were identified:Opportunities for improving SDM between prosthetists and prosthesis users include (1) clarifying key rehabilitation decisions, (2) identifying the purpose of initial communications, (3) support for balancing priorities, and (4) utilizing experience to achieve informed preferences.IMPLICATIONS FOR REHABILITATIONMany people with lower limb amputation experience poor physical function and psychosocial outcomes, which may be further compounded by under informed prosthesis-user expectations for function with a prosthesis.Shared decision-making offers an opportunity for improving realistic prosthesis-user expectations, reducing healthcare costs, and improving prosthesis-user satisfaction and adherence to care plans.Opportunities for improving shared decision-making between prosthetists and prosthesis-users include (1) clarifying key rehabilitation decisions, (2) identifying the purpose of initial communications, (3) support for balancing priorities, and (4) utilizing experience to achieve informed preferences.

Rose-ringed parakeets (Psittacula krameri) are one of the most widespread invasive avian species worldwide. This species was introduced to the island of Kaua‘i, Hawai‘i, USA, in the 1960s. The rapidly increasing population has caused substantial economic losses in the agricultural and tourism industries. We evaluated the efficacy of a roost culling program conducted by an independent contractor from March 2020 to March 2021. We estimated island-wide minimum abundance was 10,512 parakeets in January 2020 and 7,372 in April 2021. Over 30 nights of culling at four roost sites, approximately 6,030 parakeets were removed via air rifles with 4,415 (73%) confirmed via carcasses retrieval. An estimated average of 45 parakeets were removed per hour of shooter effort. The proportion of adult females removed in 2020 was 1.9 × greater when culled outside of the estimated nesting season. Of the four roosts where culling occurred, the parakeets fully abandoned three and partially abandoned one site. Of the three fully abandoned roosts, an estimated average of 29.6% of birds were culled prior to roost abandonment. The roost culling effort was conducted during the COVID-19 pandemic, when tourist numbers and foot traffic were greatly reduced. It is unknown how public perception of roost culling in public areas may impact future efforts. Findings suggest roost culling can be utilized for management of nonnative rose-ringed parakeet populations when roost size is small enough and staff size large enough to cull entire roosts in no greater than two consecutive nights (e.g., if two shooters are available for three hours per night, roost culling should only be attempted on a roost with ≤ 540 rose-ringed parakeets).

Accurate estimates and forecasts of ocean eddies in key regions such as western boundary currents are important for weather and climate, biology, navigation, and search and rescue. The dynamic nature of mesoscale eddies requires data assimilation to produce accurate eddy timings and locations in ocean model simulations. However, data assimilating models are rarely assessed below the surface due to a paucity of observations; hence it is not clear how data assimilation impacts the subsurface eddy structure. Here, we use a suite of observing system simulation experiments to show how the subsurface representation of eddies is changed within data assimilating simulations even when assimilating nearby observations. We examine in detail two possible manifestations of how the data assimilation process impacts three-dimensional eddy structure, namely, by producing overly active baroclinic instability and through inaccurate vertical mode structure. Therefore, in DA simulations, subsurface temperature structures can be too deep and too warm, particularly in dynamic eddy features. Our analyses demonstrate the need for further basic research in ocean data assimilation methodologies to improve the representation of the subsurface ocean structure.

Here we report the development and optimization of a mass spectrometry imaging (MSI) platform that combines an atmospheric-pressure matrix-assisted laser desorption/ionization platform with plasma postionization (AP-MALDI-PPI) and trapped ion mobility spectrometry (TIMS). We discuss optimal parameters for operating the source, characterize the behavior of a variety of lipid classes in positive- and negative-ion modes, and explore the capabilities for lipid imaging using murine brain tissue. The instrument generates high signal-to-noise for numerous lipid species, with mass spectra sharing many similarities to those obtained using laser postionization (MALDI-2). The system is especially well suited for detecting lipids such as phosphatidylethanolamine (PE), as well as numerous sphingolipid classes and glycerolipids. For the first time, the coupling of plasma-based postionization with ion mobility is presented, and we show the value of ion mobility for the resolution and identification of species within rich spectra that contain numerous isobaric/isomeric signals that are not resolved in the

Identifying effective classroom management strategies is important to support teachers in promoting environments conducive to learning. Effective classroom management has been demonstrated to be associated with student achievement, motivation, and fewer challenging behaviors. Given the benefits of effective classroom management strategies and providing students with self-regulation skills, the present study examined the effectiveness of a contemporary classroom behavioral management strategy in reducing off-task problem behaviors. The Raising Lions method of behavior management emphasizes: i. Immediate action responses from teachers, ii. Encourages a positive or neutral tone of voice, and iii. Allocates brief prompts for students that allow them the opportunity to exercise self-control and re-engage in classroom instruction without teacher judgment. Employing a pre-post-follow-up design, the present study examined implementation fidelity and the effectiveness of this classroom management strategy in reducing the number of off-task problem behaviors in elementary school classrooms. Classroom observations were conducted in all 18 classrooms (school-wide, including transition kindergarten through Grade 6 classrooms), with a population of primarily Latino students. One-way ANOVAs compared off-task behaviors across pre and post-intervention stages. Overall, results revealed statistically significant decreases in the number of off-task problem behaviors in 16 of the 18 classrooms. The largest impact was related to teachers not naming the behavior and instead giving a brief action prompt to trigger self-reflection and self-correction. Implications for practice and research are discussed.

Seeds are a primary source for generating plants for large-scale restoration and understanding the requirements for seed germination and establishment is fundamental to ecological restoration. Seed germination traits are central to defining the germination niche and identifying traits that may limit seedling establishment provides insights into how seeds may interact with the abiotic environment and the soil substrates specific to each restoration site and informs seed management practices. In this paper we review seed trait data derived from research to improve restoration practice across diverse ecosystems within Western Australia. We compile and evaluate seed trait data for up to 300 seed collections of 287 species that are used in restoration programmes to identify species, lifeform, and region-specific variation in seed dormancy, maximum germination, germination speed, base water potential, and germination niche breadth. Through a synthesis of this data, we outline our ecologically-guided approach to identifying key seed traits that support plant regeneration to improve the success of seed-based restoration across the biodiverse Western Australian landscape.

This article offers a summary, synthesis, and highlights of processes, surveys, summits, and discussions among members of the Society for the Study of School Psychology exploring current and historical perspectives as to a potential definition of school psychology research. After multiple revisions, the final definition is as follows: School psychology research is the systematic investigation of phenomena concerning the educational, emotional, behavioral, and social outcomes, as well as the school, family, and societal systems in which these outcomes are imbedded. It is principally concerned with socially responsive implementation and translating research into equitable practices that can lead to improved functioning of students, families, schools, and community systems that support educational and psychological services. It includes basic and applied research and theory development that focuses on a wide array of topics including, for example, prevention, intervention, assessment, diagnosis, diversity, equity and inclusion, measurement, methodology, statistics, and professional issues. This wide range of topics is informed by diverse theories arising from a broad array of disciplines and investigated by researchers from a variety of educational and scientific backgrounds. Common to all of the areas of research inquiry is a focus on enhancing student, family, and community outcomes-particularly educational and social outcomes of all students-and in building the capacity of systems (schools, communities, and other systems) to support those outcomes with a commitment to incorporating social responsiveness in the research process. The aim of this definition is to embrace the breadth of scholarship informing and advancing the science, practice, and policy relevant to the field of school psychology.

Ferroptosis is a non-apoptotic cell death mechanism characterized by the generation of lipid peroxides. While many effectors in the ferroptosis pathway have been mapped, its epitranscriptional regulation is not yet fully understood. Ferroptosis can be induced via system xCT inhibition (Class I) or GPX4 inhibition (Class II). Previous works have revealed important differences in cellular response to different ferroptosis inducers. Importantly, blocking mRNA transcription or translation appears to protect cells against Class I ferroptosis inducing agents but not Class II. In this work, we examined the impact of blocking transcription (via Actinomycin D) or translation (via Cycloheximide) on Erastin (Class I) or RSL3 (Class II) induced ferroptosis. Blocking transcription or translation protected cells against Erastin but was detrimental against RSL3. Cycloheximide led to increased levels of GSH alone or when co-treated with Erastin via the activation of the reverse transsulfuration pathway. RNA sequencing analysis revealed early activation of a strong alternative splice program before observed changes in transcription. mRNA stability analysis revealed divergent mRNA stability changes in cellular response to Erastin or RSL3. Importantly, codon optimality biases were drastically different in either condition. Our data also implicated translation repression and rate as an important determinant of the cellular response to ferroptosis inducers. Given that mRNA stability and codon usage can be influenced via the tRNA epitranscriptome, we evaluated the role of a tRNA modifying enzyme in ferroptosis stress response. Alkbh1, a tRNA demethylase, led to translation repression and increased the resistance to Erastin but made cells more sensitive to RSL3.

Recent advances in irrigation technologies have led many states to incentivize homeowners to purchase United States Environmental Protection Agency WaterSense-labeled, smart irrigation controllers. However, previous research of smart controllers has shown that their use may still result in excess water application when compared with controllers manually programmed to replace actual water loss. This study compared kentucky bluegrass (Poa pratensis) irrigation applications using three smart irrigation controllers, a conventional irrigation controller programmed according to Cooperative Extension recommendations, and the average irrigation rate of area homeowners in Utah during 2018 and 2019. Of all the controllers tested, the manually programmed controller applied water at amounts closest to the actual evapotranspiration rates; however, smart controllers applied from 30% to 63% less water than area homeowners, depending on the controller and year of the study. Kentucky bluegrass health and quality indicators—percent green cover and normalized difference vegetation indices—varied between years of the study and were lower than acceptable levels on several occasions in 2019 for three of the four controllers tested. Compared with the results of similar studies, these findings suggest that the effects of smart irrigation controllers on turfgrass health and quality may vary by location and over time.

OBJECTIVE To examine variations in timing of lower-limb amputation (LLA) across race/ethnicity and sex among older adults with a diabetic foot ulcer (DFU). It was hypothesized Black/African Americans were more likely to have LLA post-DFU earlier compared with non-Hispanic/Whites, and more men would receive LLA earlier post-DFU compared with women. RESEARCH DESIGN AND METHODS This was a retrospective cohort analysis of enrolled Medicare fee-for-service (FFS) beneficiaries with a diagnosis of DFU during the study period (2012–2017), allowing up to 5 years post-DFU. Final analytic sample contained 643,287 individuals; the subsample consisted of 68,633 individuals with LLA only. The primary outcome was mutually exclusive groups based on timing of LLA. Multinomial logistic regression was applied to assess likelihood of membership into a group post-DFU based on characteristics such as sex and race/ethnicity. RESULTS Black/African American beneficiaries had 1.98 (95% CI 1.93–2.03) times the odds of receiving an LLA within 1 year of DFU diagnosis compared with non-Hispanic/White beneficiaries relative to no amputation. Female beneficiaries had increased odds (odds ratio [OR] 1.07, 95% CI 1.02–1.11] between 1 and 3 years and OR 1.08 [95% CI 1.03–1.12] in ≥3 years) of a delayed LLA compared with men among those that underwent LLA. CONCLUSIONS Notably, these results present novel evidence on timing of LLA between racial groups and sex for Medicare FFS beneficiaries post-DFU. Results may be generalizable to individuals with Medicare FFS and DFU. Clinically more targeted, evidence-based decision making informs care decisions with opportunities to address inequities related to the social determinants of health that may lead to LLA.

Cold burns are an uncommon mechanism of burn injury in Australia, where environmental exposure to freezing temperatures is rare. Aetiological research shows that most likely causes are related to intentional injury or industrial accidents

Obscurin is a large scaffolding protein in striated muscle that maintains sarcolemmal integrity and aligns the sarcoplasmic reticulum with the underlying contractile machinery. Ankyrins are a family of adaptor proteins with some isoforms that interact with obscurin. Previous studies have examined obscurin interacting with individual ankyrins. In this study, we demonstrate that two different ankyrins interact with obscurin's carboxyl terminus via independent ankyrin-binding domains (ABDs). Using in-vitro binding assays, co-precipitation assays, and FLIM-FRET analysis, we show that obscurin interacts with small ankyrin 1.5 (sAnk1.5) and the muscle-specific ankyrin-G isoform (AnkG107). While there is no direct interaction between sAnk1.5 and AnkG107, obscurin connects the two ankyrins both in vitro and in cells. Moreover, AnkG107 recruits β-spectrin to this macromolecular protein complex and mutating obscurin's ABDs disrupts complex formation. To further characterize AnkG107 interaction with obscurin, we measure obscurin-binding to different AnkG107 isoforms expressed in the heart and find that the first obscurin-binding domain in AnkG107 principally mediates this interaction. We also find that AnkG107 does not bind to filamin-C and displays minimal binding to plectin-1 compared to obscurin. Finally, both sAnk1.5-GFP and AnkG107-CTD-RFP are targeted to the M-lines of ventricular cardiomyocytes and mutating their obscurin-binding domains disrupts the M-line localization of these ankyrin constructs. Altogether, these findings support a model in which obscurin can interact via independent binding domains with two different ankyrin protein complexes to target them to the sarcomeric M-line of ventricular cardiomyocytes.

In this study, we present preliminary results of modelling the potentially collisional atmosphere of the Jovian satellite Europa using the Direct Simulation Monte Carlo (DSMC) method [1]. In the DSMC method particular gas flows are calculated through the collision mechanics of representative atoms or molecules that are subject to binary collisions to simulate macroscopic gas dynamics.NASA's Europa Clipper mission [2] and ESA's JUpiter Icy Moons Explorer (JUICE) [3] will encounter Europa with flybys in the 2030s to sample the atmosphere of the icy moon using mass spectroscopy. Measurements with the MAss Spectrometer for Planetary EXploration (MASPEX) onboard Europa Clipper and the Neutral gas and Ion Mass spectrometer (NIM) onboard JUICE will determine the composition of Europa's exosphere and, potentially, sample the plume material. From the exosphere measurements, the chemical composition of Europa's surface could be derived, whereas plume measurements would potentially allow conclusions about the chemical conditions of Europa's subsurface ocean.Models of the collision-less exosphere for the icy moon [4, 5] have shown that Europa’s ice-sputtered atmosphere is dominated by O2 near the surface with an extended H2 corona at higher altitudes. Here, we compare the results of these studies with the DSMC model including deeper layers of Europa's collisional atmosphere.[1] Bird, G. A. (1994). Molecular gas dynamics and the direct simulation of gas flows.[2] Phillips, C. B., and Pappalardo, R. T. (2014). Eos, Transactions AGU, 95(20), 165-167.[3] Grasset, O., et al. (2013). Planetary and Space Science, 78, 1-21.[4] Vorburger, A., and Wurz, P. (2018). Icarus, 311, 135-145.[5] Vorburger, A., and Wurz, P. (2021). J. Geophys. Res. Space Phys., 126(9), e2021JA029690.

Herein we constrain the radiolytic production rate of O2 from Callisto's exposed ice patches as well as the corresponding steady-state abundance of O2 in Callisto's atmosphere. That is, by simulating the fluxes of thermal plasma and energetic particles irradiating Callisto's surface, taking into account energy deposition within the atmosphere, we determine the initial source flux of O2 to estimate the corresponding column density for Callisto's O2 component, which we compare to those suggested in the literature. This study provides constraints for Callisto's O2 atmosphere in preparation for future observations, particularly those that will be made by the JUpiter ICy moons Explorer (JUICE) and Europa Clipper spacecraft, as well as the Hubble Space Telescope (HST). Further, based on this analysis at Callisto, we can better our understanding on how the atmospheres of other icy satellites in the Solar System can evolve to their observed state.

Thioredoxin Reductase (TrxR) is a key enzyme in reactive oxygen species (ROS) detoxification and in redox regulation. Because cancer cells produce increased steady-state levels of ROS (i.e., superoxide and hydrogen peroxide), TrxR is viable target in clinical trials using the anti-rheumatic drug, auranofin (AF). To extend these observations to small cell lung cancer (SCLC), AF-mediated TrxR inhibition as well as tolerability and tumor growth inhibition was determined in a xenograft model. AF was administered intraperitoneal, daily or twice daily for 1 to 5 days in mice bearing DMS273 xenografts. AF uptake was determined by mass spectrometry of gold and inhibition of TrxR in the tumor was determined. The optimal dose was 4 mg/kg once daily resulting in 18 μM gold in the plasma and 50% inhibition of TrxR activity in DMS273 SCLC tumors. This regimen given for 14 days provided a trend for prolonged median survival from 17.5 to 22 days (p=0.058, N=20 controls, 19 AF) without causing changes in bodyweight, bone marrow toxicity, blood urea nitrogen or creatinine. These results support the hypothesis that AF is an effective inhibitor of TrxR and suggest that AF could be used as an adjuvant in radio-chemotherapy protocols to enhance therapeutic efficacy.

Objective:High cholesterol levels in pancreatic β-cells cause oxidative stress and decrease insulin secretion. β-cells can internalize apolipoprotein (apo) A-I, which increases insulin secretion. This study asks whether internalization of apoA-I improves β-cell insulin secretion by reducing oxidative stress.Approach:Ins-1E cells were cholesterol-loaded by incubation with cholesterol-methyl-β-cyclodextrin. Insulin secretion in the presence of 2.8 or 25 mM glucose was quantified by radioimmunoassay. Internalization of fluorescently labelled apoA-I by β-cells was monitored by flow cytometry. The effects of apoA-I internalization on β-cell gene expression was evaluated by RNA sequencing. ApoA-I binding partners on the β-cell surface were identified by mass spectrometry. Mitochondrial oxidative stress was quantified in β-cells and isolated islets with MitoSOX and confocal microscopy.Results:An F1-ATPase β-subunit on the β-cell surface was identified as the main apoA-I binding partner. β-cell internalization of apoA-I was time-, concentration-, temperature-, cholesterol- and F1-ATPase β-subunit-dependent. β-cells with internalized apoA-I (apoA-I+cells) had higher cholesterol and cell surface F1-ATPase β-subunit levels than β-cells without internalized apoA-I (apoA-I−cells). The internalized apoA-I co-localized with mitochondria and was associated with reduced oxidative stress and increased insulin secretion. The ATPase inhibitory factor 1, IF1, attenuated apoA-I internalization and increased oxidative stress in Ins-1E β-cells and isolated mouse islets. Differentially expressed genes in apoA-I+and apoA-I−Ins-1E cells were related to protein synthesis, the unfolded protein response, insulin secretion and mitochondrial function.Conclusions:These results establish that β-cells are functionally heterogeneous and apoA-I restores insulin secretion in β-cells with elevated cholesterol levels by improving mitochondrial redox balance.

Although many dangerous effects of smoking e-cigarettes on lungs have come to light, limited and only qualitative efforts have been made to analyze the impact of e-cigarettes on the human heart directly. In this study, we experimentally determined e-liquid cardiotoxicity in both healthy cells and cells with long QT syndrome by treating healthy and diseased human cardiomyocytes with e-liquids with varying nicotine concentrations. These cardiomyocytes were generated from human induced pluripotent stem cells. The cardiomyocytes were divided into 5 groups, a control group and 4 test groups, each treated with e-liquid containing varying amounts of nicotine between 0% and 70%. The cells’ biological indicators such as heart rate, pulse pressure, essential protein concentration, and metabolic activity, were measured and characterized using three different functional assays: contractility, Western blot, and viability, and tracked over 4 days. The results demonstrated that acute exposure to e-liquid led to tachycardia, hypertension, decreased protein levels, and cell death. The rate of cardiotoxicity increases with higher nicotine concentrations. The basal fluid also showed non-negligible toxicity. Under identical conditions, the functionality of the diseased heart cells declined at a faster rate compared to healthy cells. Overall, this work establishes the harmful physiological effects of e-cigarettes on the human heart quantitatively.

BackgroundWest Nile virus (WNV) is the leading cause of mosquito-borne disease in humans in the United States. Since the introduction of the disease in 1999, incidence levels have stabilized in many regions, allowing for analysis of climate conditions that shape the spatial structure of disease incidence.ObjectivesOur goal was to identify the seasonal climate variables that influence the spatial extent and magnitude of WNV incidence in humans.MethodsWe developed a predictive model of contemporary mean annual WNV incidence using U.S. county-level case reports from 2005 to 2019 and seasonally averaged climate variables. We used a random forest model that had an out-of-sample model performance of R2=0.61.ResultsOur model accurately captured the V-shaped area of higher WNV incidence that extends from states on the Canadian border south through the middle of the Great Plains. It also captured a region of moderate WNV incidence in the southern Mississippi Valley. The highest levels of WNV incidence were in regions with dry and cold winters and wet and mild summers. The random forest model classified counties with average winter precipitation levels

Supplementary text

Purpose:PIK3CA mutations are common in breast cancer and promote tumor progression and treatment resistance. We conducted a phase I/II trial of alpelisib (α-specific PI3K inhibitor) plus nab-paclitaxel in patients with HER2-negative metastatic breast cancer (MBC).Patients and Methods:Eligible patients had HER2-negative MBC with any number of prior chemotherapies. Phase I was 3+3 dose-escalation design with three dose levels of alpelisib (250, 300, and 350 mg) daily plus nab-paclitaxel 100 mg/m2 administered on days 1, 8, and 15 every 28 days. Phase II was according to Simon's two-stage design. PIK3CA mutations in tumor/circulating tumor DNA (ctDNA) were assessed. Primary endpoints were recommended phase II dose (RP2D) and objective response rate (ORR). Additional endpoints included safety, pharmacokinetics, progression-free survival (PFS), and association of PIK3CA mutation with outcomes.Results:A total of 43 patients were enrolled (phase I, n = 13 and phase II, n = 30). A total of 84% had visceral disease and 84% had prior taxane. No dose-limiting toxicities occurred in phase I. RP2D was alpelisib 350 mg daily plus nab-paclitaxel 100 mg/m2 on days 1, 8, and 15. Hyperglycemia (grade 3, 26% and grade 4, 0%), neutropenia (grade 3, 23% and grade 4, 7%), diarrhea (grade 3, 5% and grade 4, 0%), and rash (grade 3, 7% and grade 4, 0%) were the most common adverse events. Among 42 evaluable patients, ORR was 59% (complete response, 7% and partial response, 52%), 21% of whom had response lasting >12 months; median PFS was 8.7 months. A total of 40% of patients demonstrated tumor and/or ctDNA PIK3CA mutation; patients with tumor/ctDNA mutation demonstrated better PFS compared with those without mutation (11.9 vs. 7.5 months; HR, 0.44; P = 0.027). Patients with normal metabolic status had longer PFS compared with prediabetic/diabetic patients (12 vs. 7.5 months; P = 0.014). No pharmacokinetics interactions were detected.Conclusions:The alpelisib plus nab-paclitaxel combination was well tolerated and shows encouraging efficacy, especially in patients with PIK3CA-mutated tumor/ctDNA. The impact of metabolic status on response to this combination merits further investigation.

Kaplan-Meier estimates of progression-free survival by baseline metabolic status

Gene expression signatures by PIK3CA mutation status

Kaplan-Meier estimates of overall survival

Image-based profiling has emerged as a powerful technology for various steps in basic biological and pharmaceutical discovery, but the community has lacked a large, public reference set of data from chemical and genetic perturbations. Here we present data generated by the Joint Undertaking for Morphological Profiling (JUMP)-Cell Painting Consortium, a collaboration between 10 pharmaceutical companies, six supporting technology companies, and two non-profit partners. When completed, the dataset will contain images and profiles from the Cell Painting assay for over 116,750 unique compounds, over-expression of 12,602 genes, and knockout of 7,975 genes using CRISPR-Cas9, all in human osteosarcoma cells (U2OS). The dataset is estimated to be 115 TB in size and capturing 1.6 billion cells and their single-cell profiles. File quality control and upload is underway and will be completed over the coming months at the Cell Painting Gallery:https://registry.opendata.aws/cellpainting-gallery. A portal to visualize a subset of the data is available athttps://phenaid.ardigen.com/jumpcpexplorer/.

A dynamical understanding of the physical process of surface gravity wave breaking remains an unresolved problem in fluid dynamics. Conceptually, breaking can be described by inception and onset, where breaking inception is the initiation of unknown irreversible processes within a wave crest that precede the visible manifestation of breaking onset. In the search for an energetic indicator of breaking inception, we use an ensemble of non-breaking and breaking crests evolving within unsteady wave packets simulated in a numerical wave tank to investigate the evolution of each term in the kinetic energy balance equation. We observe that breaking onset is preceded by around one quarter of a wave period by a rapid increase in the rate of convergence of kinetic energy that triggers an irreversible acceleration of the kinetic energy growth rate. This energetic signature, which is present only for crests that subsequently break, arises when the kinetic energy growth rate exceeds a critical threshold. At this point the additional kinetic energy convergence cannot be offset by converting excess kinetic energy to potential energy or by dissipation through friction. Our results suggest that the ratio of the leading terms of the kinetic energy balance equation at the time of this energetic signature is proportional to the strength of the breaking crest. Hence this energetic inception point both predicts the occurrence of breaking onset and indicates the strength of the breaking event., Comment: Submitted for publication to the Journal of Fluid Mechanics

Advances in peroxidase- and biotin ligase-mediated signal amplification have enabled high-resolution subcellular mapping of endogenous RNA localization and protein-protein interactions. Application of these technologies has been limited to RNA and proteins because of the reactive groups required for biotinylation in each context. Here we report several novel methods for proximity biotinylation of exogenous oligodeoxyribonucleotides by application of well-established and convenient enzymatic tools. We describe approaches using simple and efficient conjugation chemistries to modify deoxyribonucleotides with “antennae” sensitive to phenoxy radical or biotinoyl-5’-adenylate. In addition, we report chemical details of a previously undescribed adduct between tryptophan and a phenoxy radical group. These developments have potential application in the selection of exogenous nucleic acids capable of unaided entry into living cells.

Triple-negative breast cancer (TNBC) patients with residual disease (RD) after neoadjuvant systemic therapy (NAST) are at high risk for recurrence. Biomarkers to risk-stratify patients with RD could help individualize adjuvant therapy and inform future adjuvant therapy trials. We aim to investigate the impact of circulating tumor DNA (ctDNA) status and residual cancer burden (RCB) class on outcomes in TNBC patients with RD. We analyze end-of-treatment ctDNA status in 80 TNBC patients with residual disease who are enrolled in a prospective multisite registry. Among 80 patients, 33% are ctDNA positive (ctDNA+) and RCB class distribution is RCB-I = 26%, RCB-II = 49%, RCB-III = 18% and 7% unknown. ctDNA status is associated with RCB status, with 14%, 31%, and 57% of patients within RCB-I, -II, and -III classes demonstrating ctDNA+ status (P = 0.028). ctDNA+ status is associated with inferior 3-year EFS (48% vs. 82%, P P = 0.002). ctDNA+ status predicts inferior 3-year EFS among RCB-II patients (65% vs. 87%, P = 0.044) and shows a trend for inferior EFS among RCB-III patients (13% vs. 40%, P = 0.081). On multivariate analysis accounting for T stage and nodal status, RCB class and ctDNA status independently predict EFS (HR = 5.16, P = 0.016 for RCB class; HR = 3.71, P = 0.020 for ctDNA status). End-of-treatment ctDNA is detectable in one-third of TNBC patients with residual disease after NAST. ctDNA status and RCB are independently prognostic in this setting.

Intramuros, also known as the "Walled City," is one of the Philippines' pride and a top tourist spot in Manila. If marketed well, more people will go down memory lane and learn about this must-see destination. One will venture into its culture and learn the history within the walls. To make use of this advantage, a platform is effective. Thus, social media impact is seen nowadays as an effective way to create an impact in the tourism industry. This study aims to explore how social media affected the tourism growth in Intramuros as perceived by tourists from National Capital Region.This descriptive study utilized snowball sampling resulting in 400 respondents from the National Capital Region, aged 18 and above, who visited Intramuros. This study is anchored on Social Media Integration Theory which presents how social media has been utilized for delivering information. The findings revealed that social media has a significant part in Intramuros' tourism growth. Thus, it serves as a popular platform among visitors to research trips and shares their travel's most significant memories. Reviews and feedback help determine the needed improvement of Intramuros, and social media serves as a cost-effective way to reach target audiences. The walled city's cultural, historical, and religious benefits are some of the factors that influence the tourist to visit Intramuros.It concluded that social media is a meaningful marketing tool; therefore, it will lead to an excellent platform to attract potential visitors. Tourists' posted data seem to be a potential, credible, reliable source to represent the historic walled city of Intramuros.

Early mobility, functional independence, and ambulation are associated benefits after lower limb amputation (LLA), whereas an increased risk of clinical complications is associated with no prosthesis.To describe time to prosthesis receipt after amputation and to assess the impact of patient demographic and health factors on the rate of prosthesis receipt within 12 months post LLA.A retrospective cohort analysis using commercial administrative claims data. Kaplan-Meier and Cox proportional-hazards models were applied to assess time to prosthesis receipt.Watson/Truven administrative database 2014-2016.Adults aged 18-64 years with LLA who maintained their current insurance enrollment for 12 months after amputation.Independent variables included diabetes/vascular disease status, amputation level, age, gender, and region.Prosthesis receipt was defined based on the presence of codes billed for prosthesis services. Time was measured in days from date of amputation surgery.Among the sample, 510 individuals maintained insurance enrollment for 12 months after amputation, of which 443 individuals received a prosthesis within that period (79% below knee and 21% above knee). The adjusted average rate of time to prosthesis receipt was 138 (95% confidence interval [CI]: 113-185) days. Individuals with diabetes/vascular disease were 22% (hazard ratio: 1.22, 95% CI: 1.02-1.49) more likely to receive a prosthesis earlier than individuals without diabetes/vascular disease and women received a prosthesis later than men at 141 (95% CI: 126-162) days versus 106 (95% CI: 96-119) days, respectively.This study expands the understanding of factors that influence the likelihood of receiving a prosthesis along with the timing of prosthesis receipt after LLA among commercially insured adults. At least half of this sample received a prosthesis within 5 months or less. Disparities in timing and access to a prosthesis based on amputation level and gender were noted; future efforts are needed to address these issues.

Liver fibrosis progression in chronic liver disease leads to cirrhosis, liver failure, or hepatocellular carcinoma and often ends in liver transplantation. Even with an increased understanding of liver fibrogenesis and many attempts to generate therapeutics specifically targeting fibrosis, there is no approved treatment for liver fibrosis. To further understand and characterize the driving mechanisms of liver fibrosis, we developed a high-throughput genome-wide CRISPR/Cas9 screening platform to identify hepatic stellate cell (HSC)-derived mediators of transforming growth factor (TGF)-β-induced liver fibrosis. The functional genomics phenotypic screening platform described here revealed the novel biology of TGF-β-induced fibrogenesis and potential drug targets for liver fibrosis.

Conceptual foundations inform our professional activities as practitioners and scholars. This paper describes a theoretical framework to inform and advance scientific and applied work, recognizing the numerous factors influencing the trajectory of human development and the imperative of transforming the traditional “ways of being” in the field of education and applied psychology through the lens of critical theories. The central tenets of a critical, transactional, ecological, and developmental perspective are grounded in theories from developmental scholars including the works of Sameroff (e.g., transactional), Bronfenbrenner (e.g., ecological), Bowlby (e.g., relationships), and the Frankfurt school (e.g., critical theory). An emphasis is placed on the dynamic and reciprocal transactions between the individual and their environment across time and the social complexities brought by the intersection of these ongoing, dynamic, and reciprocal social relations. This paper also includes a description of empirical support informing these conceptual foundations with special attention to longitudinal studies demonstrating various influential factors across diverse developmental trajectories. Also included is a discussion of, implications informing prevention and intervention efforts to promote academic and socialemotional learning for all children, future research directions, and advocacy stance to promote shift and change to interrupt the long-standing social and educational injustices in the field of education.

Purpose BRCA1/2 pathogenic variant (PV) mutations confer radiation sensitivity preclinically, but there are limited data regarding breast cancer outcomes after radiation therapy (RT) among patients with documented BRCA1/2 PV mutations versus no PV mutations. Methods and Materials This retrospective cohort study included women with clinical stage I-III breast cancer who received definitive surgery and RT and underwent BRCA1/2 genetic evaluation at the The University of Texas MD Anderson Cancer Center. Rates of locoregional recurrence (LRR), disease-specific death (DSD), toxicities, and second cancers were compared by BRCA1/2 PV status. Results Of the 2213 women who underwent BRCA1/2 testing, 63% self-reported their race as White, 13.6% as Black/African American, 17.6% as Hispanic, and 5.8% as Asian/American Indian/Alaska Native; 124 had BRCA1 and 100 had BRCA2 mutations; and 1394 (63%) received regional nodal RT. The median follow-up time for all patients was 7.4 years (95% confidence interval [CI], 7.1-7.7 years). No differences were found between the groups with and without BRCA1/2 PV mutations in 10-year cumulative incidences of LRR (with mutations: 11.6% [95% CI, 7.0%-17.6%]; without mutations: 6.6% [95% CI, 5.3%-8.0%]; P = .466) and DSD (with mutations: 12.3% [95% CI, 8.0%-17.7%]; without mutations: 13.8% [95% CI, 12.0%-15.8%]; P = .716). On multivariable analysis, BRCA1/2 status was not associated with LRR or DSD, but Black/African American patients (P = .036) and Asians/American Indians/Alaska Native patients (P = .002) were at higher risk of LRR compared with White patients, and Black/African American patients were at higher risk of DSD versus White patients (P = .004). No in-field, nonbreast second cancers were observed in the BRCA1/2 PV group. Rates of acute and late grade ≥3 radiation-related toxicity in the BCRA1/2 PV group were 5.4% (n = 12) and 0.4% (n = 1), respectively. Conclusions Oncologic outcomes in a diverse cohort of patients with breast cancer who had a germline BRCA1/2 PV mutation and were treated with RT were similar to those of patients with no mutation, supporting the use of RT according to standard indications in patients with a germline BRCA1/2 PV mutation.

Individuals with lower limb amputation have a high incidence of falls. Above-the-knee amputation and diabetes/vascular disease are both risk factors for falls. Microprocessor knee (MPK) technology may reduce falls in this population. The objective was to determine the association between MPKs and reduced injurious falls. A retrospective analysis of injurious falls within a large, national outcomes database was conducted. Inclusion was limited to adult K3 ambulators with unilateral, transfemoral, or knee disarticulation amputation due to diabetes/vascular disease. There were 744 out of 881 individuals that did not receive an MPK. Results showed that 16.3% of non-MPK users experienced an injurious fall compared to 7.3% of MPK users (

Invasive mice (Mus spp.) can negatively impact island species and ecosystems. Because fewer island rodent eradications have been attempted for mice compared to rats (Rattus spp.), less is known about efficacy and palatability of rodenticide baits for mouse eradications. We performed a series of bait acceptance and efficacy cage trials using a standard formulation of brodifacoum-based rodenticide on wild-caught mice from Sand Island, Midway Atoll, to help inform a proposed eradication there. Mice were offered ad libitum brodifacoum pellets along with various alternative food sources, and a “no choice” treatment group received only bait pellets. Mortality in the no choice trial was 100%; however, when offered alternative foods, mice preferred the alternative diets to the bait, leading to low mortality (40%). Because there was concern that the bittering agent Bitrex® in the formulation may have reduced palatability, we conducted a subsequent trial comparing brodifacoum bait with and without Bitrex. Mortality in the with-Bitrex treatment group was slightly higher, indicating that the bittering agent was not likely responsible for low efficacy. Laboratory trials cannot account for the numerous environmental and behavioral factors that influence bait acceptance nor replicate the true availability of alternative food sources in the environment, so low efficacy results from these trials should be interpreted cautiously and not necessarily as a measure of the likelihood of success or failure of a proposed eradication.