Implementation and sustainability of public health approaches for mental health and wellbeing in schools

Aims: With increasing prevalence of emotional difficulties in children and young people in England, there is a growing emphasis on prevention programs delivered in school settings. The Education for Wellbeing program is working with around 400 schools in England to evaluate five universal mental health and wellbeing interventions Here, we present an overview of the Education for Wellbeing program and describe patterns across different school settings in barriers and facilitators to sustaining intervention delivery Methods: This study draws on interviews with staff and pupils from eight schools over multiple timepoints, including during the COVID- 19 pandemic. Qualitative methods were used to cluster schools with similar 'journeys' over a three-year period in terms of staff members' experiences of intervention delivery and perceived barriers and facilitators to sustaining delivery Results: The analysis demonstrated patterns in schools' journeys over time, each underpinned by a range of barriers and facilitators to the sustainability of the interventions. Four clusters of schools were identified each representing one overarching pattern: 'Spreading and embedding', 'Trialled and moved on', 'Everything's changed', and 'Built into the curriculum for now' Conclusion(s): The variety in schools' experiences highlights the complexity of both school settings and the process of implementing and sustaining interventions. These findings suggest that the environment and conditions into which a public health intervention is placed may be as, if not more, important than the intervention itself, and provoke important questions regarding future research and intervention development.

Resilience of U.S. cattle and beef sectors: lessons from COVID-19. (Special Issue: COVID-19 impacts on selected commodities.)

The upheaval wrought on the U.S. beef industry by the global COVID-19 pandemic carried with it several lessons that might help improve resiliency should there be a reoccurrence. First, the futures market for fed cattle fell well before cash prices, which sent a signal to market cattle early, and those who did so benefited. Second, the decline in futures anticipated the closure of slaughter plants and provided an opportunity to purchase and store beef primals in anticipation of future scarcity. Third, the beef industry has ways of slowing or stopping the pipeline of animals destined for feed yards and can "store" these animals in background feeding facilities or on pasture or rangeland. Producers who waited to sell feeder cattle benefited from higher feeder cattle prices once the processing facilities reopened. Fourth, cow slaughter plants responded to the pandemic and subsequent scarcity of labor much better than large fed-cattle plants. Cow plants are not as sophisticated and complex as fed-cattle plants. This relative simplicity may help explain the superior performance of these plants during the crisis. Sixth, the academic work on the value of building smaller plants as a response against concentration provides mixed results-these plants require more labor per animal and can be even more susceptible to labor scarcity. Seventh, the observed increase in boxed beef prices, even as fed cattle prices fell, demonstrates the risk-mitigating impact of producer ownership of downstream activities in the value chain.

Resilience of U.S. Cattle and Beef Sectors: Lessons from COVID-19

The upheaval wrought on the U.S. beef industry by the global COVID-19 pandemic carried with it several lessons that might help improve resiliency should there be a reoccurrence. First, the futures market for fed cattle fell well before cash prices, which sent a signal to market cattle early, and those who did so benefited. Second, the decline in futures anticipated the closure of slaughter plants and provided an opportunity to purchase and store beef primals in anticipation of future scarcity. Third, the beef industry has ways of slowing or stopping the pipeline of animals destined for feed yards and can "store"these animals in background feeding facilities or on pasture or rangeland. Producers who waited to sell feeder cattle benefited from higher feeder cattle prices once the processing facilities reopened. Fourth, cow slaughter plants responded to the pandemic and subsequent scarcity of labor much better than large fed-cattle plants. Cow plants are not as sophisticated and complex as fed-cattle plants. This relative simplicity may help explain the superior performance of these plants during the crisis. Sixth, the academic work on the value of building smaller plants as a response against concentration provides mixed results - these plants require more labor per animal and can be even more susceptible to labor scarcity. Seventh, the observed increase in boxed beef prices, even as fed cattle prices fell, demonstrates the risk-mitigating impact of producer ownership of downstream activities in the value chain. © 2022 the author(s), published by De Gruyter, Berlin/Boston 2022.

Researching adolescent mental health during a pandemic: A registered report of a natural experiment within two pragmatic school-based trials

Aims: Research on the impact of the pandemic on adolescent mental health is limited by several methodological factors, such as the lack of pre-pandemic scores for comparison, control for developmental change, and wider geographic and social representation. Empirical evidence which can potentially causally attribute changes to the pandemic remains therefore limited. The presentation will focus on how a natural experiment within two ongoing school-based cluster randomised pragmatic trials was used to overcome such challenges to investigate whether and to which degree differences in adolescents' mental health could be attributable to the pandemic. Method(s): The multi-partner structure of the trials separated relevant aspects of the data collection from the research team, therefore the analyses could be prospectively registered. The necessity of two recruitment cohorts led to one cohort of schools being assessed in Sept-Oct 2018 (baseline) and Jan-Mar 2020 (1-year follow-up);and the other cohort in Sept-Oct 2019 and Feb-Apr 2021, respectively. Participants in the first cohort (90 schools, N = 6419) acted as unexposed controls, and participants in the second cohort (88 schools, N = 5031) were exposed to the COVID-19 pandemic between assessments. The outcomes were depressive symptoms (Short Mood and Feelings Questionnaire, primary outcome), externalising difficulties (Me and My Feelings questionnaire), and life satisfaction (Huebner Life Satisfaction Scale). Analyses were conducted with random intercept regression models with exposure to the COVID-19 pandemic as focal predictor while controlling for baseline scores and individual-and school-level covariates. Result(s): The primary outcome analysis showed higher levels of depressive symptoms (adjusted d = 0.11). For the secondary outcomes, life satisfaction scores were lower (adjusted d = 0.12), and no differences were detected for externalising difficulties (adjusted d = 0.01). The analyses further indicated potential differential effects based on socio-economic background and gender. The results were robust in sensitivity analyses exploring the impact of missing data, drop-out, and comparability of participants in both cohorts. Conclusion(s): A multi-site and interdisciplinary approach to trial planning laid the foundation for a responsive research structure that in addition to work on high-quality evidence for school-based universal interventions provided evidence that the COVID-19 pandemic increased adolescent depressive symptoms and potentially decreased life satisfaction.

Inhibition of estrogen receptor signaling as a strategy for radiosensitization of ER+ breast cancers

Purpose: The estrogen receptor (ER) is expressed in over 80% of breast tumors and has been shown to be a significant driver of breast cancer (BC) pathogenesis and therefore a target of effective first-line therapies. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of ER+ BC, the effect of ET on tumor radiosensitization remains unclear, with concerns it may be radioprotective based on G1 cell arrest with ET treatment. Here we assessed the efficacy and mechanism of ER-mediated radiosensitization using various pharmacologic approaches in ER+ BC. Methods: Radiosensitization with ER inhibitors (tamoxifen [TAM], fulvestrant [FULV], AZD9496) was assessed using clonogenic survival assays. DNA damage was assessed by the neutral comet assay. Efficiency of homologous recombination (HR) or non-homologous end joining (NHEJ) as well as changes in cell cycle, apoptosis, and senescence were assessed. The efficacy of TAM with RT in vivo was assessed with an MCF-7 xenograft model. Results: The selective estrogen receptor modulator TAM radiosensitized ER+ MCF-7 (enhancement ratio [enhR]: 1.14-1.50) and T47D (enhR: 1.33-1.60) cells but not ER-negative SUM-159 cells (enhR: 0.99-1.02). The selective estrogen receptor degrader (SERD) FULV had similar radiosensitization effects in MCF-7 (enhR: 1.33-1.76) and T47D cells (enhR: 0.97-2.81) with no radiosensitization observed in SUM-159 cells (enhR: 1.01-1.03). The novel oral SERD AZD9496 radiosensitized MCF-7 cells (enhR: 1.36-1.56). MCF-7 cells treated with TAM and RT had an increase in dsDNA breaks compared to RT alone as measured by the comet assay (p<0.05) and a decrease in NHEJ-mediated repair with TAM (p<0.05). No changes were observed in HR-mediated repair by Rad51 foci or a reporter (p=NS). RT alone and in combination with TAM or FULV induced similar levels of cell cycle arrest, suggesting that radiosensitization with the combination therapy is cell-cycle independent. There were no significant changes in apoptosis with TAM, FULV, RT, or the combination (p=NS). Although TAM or FULV did induce senescence, ET with RT increased senescence induction (p<0.05). In vivo, combination RT and TAM led to a significant delay in days to tumor doubling (control: 17, TAM: 40, RT: 32, TAM+RT: undefined;p<0.0001), and a significant difference in tumor growth between mice treated with TAM or RT alone compared combination treatment, with no increased toxicities or skin lesions from the combination treatment. Conclusion: Our data suggest that TAM, FULV, or AZD9496 can radiosensitize ER+ breast tumors, and these agents with RT may be more effective for radiosensitization. This work also supports further clinical investigation of the timing of RT for patients receiving ET, including using ET during RT, especially as initiating ET prior to RT has been increasingly utilized as a bridging therapy followed by concurrent ET+RT during the COVID-19 pandemic.

Research update: optimizing the use of American rescue funds in pork and beef

The American Rescue Funds Program seeks improvements to infrastructure, capacity, and diversification in meat and poultry processing, with clear prioritization of increased competition via small- and medium-sized processing facilities. The need to euthanize animals at a time when retailers were rationing meat sales was one of several examples of market failures during the COVID-19 pandemic, and the U.S. Department of Agriculture estimated the disruptions to agricultural meat, poultry, and egg production at $15 billion based on CFAP and CFAP2 payments. Marani et al. (2021) estimate the probability of a repeat event at 1% to 2% per year, justifying the use of these public funds to add surplus capacity and infrastructure to mitigate disruptions in case of recurrence. Economics of scale are modest beyond slaughter of more than 125 head per hour in beef plants and 2,000 head per day in pork plants (Duewer and Nelson, 1991;Ollinger, MacDonald, and Madison, 2005). Dozens of such "medium-sized" U.S. pork and beef processing plants have survived since 2000, typically relying upon niche market connections. Given historic processing plant construction costs for medium-sized plants (Aherin, 333333 2021) and an assumed 20% USDA grant to incentivize construction, a $100 million expenditure on each of the beef and pork plants creates an opportunity to add as much as 5% additional capacity for each species, easing current capacity as the industries prepare for local and export growth. Whether producer-ownership of capacity can generate stability and additional benefits in the supply chains is of key interest. Models of producer ownership-including cooperatives and carefully structured LLCs-allow livestock producers to capture processing margins and remove some of the price uncertainty around live animal prices to the plant and producer. It follows, too, that producer-ownership can therefore reduce the ability of existing larger plants to poach supply from medium-sized plants during the crucial startup phase and ensure that plants run at optimum capacity. A significant portion of the additional capacity added to the pork industry in the last 15 years exhibited some form of producer ownership. Anecdotally, the pork and beef sectors may be moving away from commodity production and into systems that maintain animal identity from farm to consumer. Producers have an opportunity to capitalize on this shift by collectively investing in medium-sized plants with the ability to preserve identity and be more responsive to evolving consumer preferences. An overarching concern is of the need to maintain capacity into the future and the potential of existing packers to acquire this subsidized capacity should medium sized processing fail.

Endocrine therapy treatment radiosensitizes estrogen receptor-positive breast cancers

Purpose: Estrogen receptor (ER) expression is present in over 80% of breast tumors and has been shown to be a significant driver of breast cancer (BC) pathogenesis and therefore a target of first-line therapies for ER-positive (ER+) BC patients. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of ER+ BC, the sequencing of therapy and the effect of ET on tumor radiosensitization remain unclear. Recently, this question has become much more clinically relevant when many physicians started offering ET as a bridging strategy to surgery and RT during the COVID-19 pandemic. Here we assessed the efficacy and mechanism of ER inhibition in ER+ BC in combination with RT in preclinical models. Methods: Clonogenic survival assays were used to assess radiosensitization. Inhibition of ER signaling was accomplished by treating ER+ MCF-7 and T47D cells with the selective ER modulator (SERM), tamoxifen, or the selective ER degrader (SERD), fulvestrant. The ER-negative SUM-159 cells were used as a negative control. DNA damage was assessed by the neutral comet assay. Efficiency of homologous recombination (HR) was measured by Rad51 foci or a GFP reporter system. Non-homologous end joining (NHEJ) efficiency was assessed with a pEYFP reporter. Cell cycle effects were measured using flow cytometry with propidium iodide (PI) staining. Apoptosis was assessed by annexin V/PI via flow Scytometry. Senescence was measured using β-galactosidase staining. Western blotting was used to quantify expression of proteins and phospho-proteins involved in cell cycle and apoptosis. An MCF-7 xenograft model was used to assess the efficacy of tamoxifen with RT in vivo. Synergy was determined using the fractional tumor volume (FTV) method. Results: ER inhibition with tamoxifen radiosensitized ER+ MCF-7 (10-250 nM, enhR: 1.14-1.50) and T47D (500 nM-2.0 μ M, enhR: 1.33-1.60) cells but not ER-negative SUM-159 cells (500 nM-2.0 μ M, enhR: 0.99-1.02). ER degradation with fulvestrant had similar radiosensitization effects in MCF-7 (1-25 nM, enhR: 1.33-1.76) and T47D cells (0.5-5 nM, enhR: 0.97-2.81) with no radiosensitization observed in SUM-159 cells (1-25 nM, enhR: 1.01-1.03). MCF-7 cells treated with 500 nM tamoxifen and 4 Gy RT had an increase in dsDNA breaks compared to RT alone as measured by the comet assay (p<0.05), and there was a decrease in NHEJ-mediated repair with tamoxifen treatment (p<0.05). No changes were observed in HR-mediated repair by Rad51 foci or an HR reporter (p=NS). RT alone and in combination with tamoxifen and fulvestrant induced similar levels of cell cycle arrest, suggesting that radiosensitization with the combination therapy is a cell-cycle independent effect. In addition, there were no significant changes in apoptosis in MCF-7 or T47D cells with endocrine therapy, RT, or the combination (p=NS). Although treatment with ET did induce senescence in ER+ MCF-7 and T47D cells, the combination treatment of ET with RT induced senescence to a much greater level suggesting this mechanism may contribute to radiosensitization (p<0.05). In vivo, combination RT and tamoxifen led to a significant delay in time to tumor doubling (17 days in control, 40 days with tamoxifen alone, 32 days with RT alone, and undefined with combination;p<0.0001) and a significant difference in tumor growth between mice treated with tamoxifen or RT alone compared to mice treated with tamoxifen and RT with synergy noted with combination treatment (FTV 1.297). Conclusion: Our data suggest that ET can radiosensitize ER+ breast tumors, and ET with RT may be more effective for radiosensitization. Ongoing studies will address concurrent versus sequential ET with RT. This work also supports further clinical investigation of the timing of RT for patients receiving ET, especially as ET prior to RT is increasingly used as a bridging therapy during the COVID-19 pandemic.

Inhibition of Estrogen Receptor Signaling as a Strategy for Radiosensitization of ER+ Breast Cancers

Purpose/Objective(s): Estrogen receptor (ER) expression is present in over 80% of breast tumors and has been shown to be a significant driver of tumor initiation and progression and therefore a target of first-line therapies for ER-positive (ER+) breast cancer (BC) patients. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of women with ER+ BC, the sequencing of therapy and the effect of ET on tumor radiosensitization remains unclear. Here we assessed the efficacy and mechanism of ER inhibition in ER+ BC in combination with RT in multiple preclinical models. Materials/Methods: Clonogenic survival assays were used to assess radiosensitization. ER inhibition was accomplished with tamoxifen and the selective ER degrader (SERD), fulvestrant, in ER+ MCF-7 and T47D cells or ER-negative (ER-) SUM-159 cells. DNA damage was assessed by yH2AX foci. Efficiency of homologous recombination (HR) or non-homologous end joining (NHEJ) was measured by RAD51 foci or using a pYFP reporter, respectively. Cell cycle effects were measured using flow cytometry with propidium iodide (PI) staining. Apoptosis was assessed by annexin V/PI via flow cytometry. Western blotting was used to quantify protein expression. An MCF-7 xenograft model was used to assess the efficacy of tamoxifen with RT in vivo where mice were pretreated with tamoxifen for one or six days prior to starting RT. Synergy was determined using the fractional tumor volume method. Results: One-hour pretreatment with tamoxifen prior to RT resulted in radiosensitization of ER+ MCF-7 (enhR: 1.14-1.50) and T47D (enhR: 1.33-1.60) cells but not ER- SUM-159 cells (enhR: 0.99-1.02). MCF-7 and T47D cells treated with tamoxifen and RT did not alter the kinetics of dsDNA break repair as measured by yH2AX foci (P > 0.05) but demonstrated a decrease in NHEJ-mediated repair (P < 0.05). No changes were observed in HR-mediated repair by Rad51 foci (P > 0.05). While cell cycle arrest was induced at 24 hours after RT, no changes were observed with tamoxifen treatment in combination with RT. In addition, there were no significant changes in apoptosis in MCF-7 or T47D cells with treatment of tamoxifen, RT, or the combination (P > 0.05). In vivo, an MCF-7 xenograft study demonstrated a significant delay in time to tumor doubling (17 days in control, 40 days with tamoxifen, 32 days with RT, and undefined with combination;P < 0.0001) and a significant difference in tumor growth between mice treated with tamoxifen or RT alone compared to mice treated with tamoxifen and RT with synergy noted with combination treatment. Conclusion: Our data suggest that endocrine therapies may be effectively used to radiosensitize ER+ breast tumors. This work also supports further clinical investigation of the timing of RT for patients receiving ET as concurrent use may be more effective than sequential, especially as ET prior to RT is increasingly used as a bridging therapy during the COVID pandemic.

Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) variants govern transmissibility, responsiveness to vaccination, and disease severity. In a screen for new models of SARS-CoV-2 infection, we identify human H522 lung adenocarcinoma cells as naturally permissive to SARS-CoV-2 infection despite complete absence of angiotensin-converting enzyme 2 (ACE2) expression. Remarkably, H522 infection requires the E484D S variant; viruses expressing wild-type S are not infectious. Anti-S monoclonal antibodies differentially neutralize SARS-CoV-2 E484D S in H522 cells as compared to ACE2-expressing cells. Sera from vaccinated individuals block this alternative entry mechanism, whereas convalescent sera are less effective. Although the H522 receptor remains unknown, depletion of surface heparan sulfates block H522 infection. Temporally resolved transcriptomic and proteomic profiling reveal alterations in cell cycle and the antiviral host cell response, including MDA5-dependent activation of type I interferon signaling. These findings establish an alternative SARS-CoV-2 host cell receptor for the E484D SARS-CoV-2 variant, which may impact tropism of SARS-CoV-2 and consequently human disease pathogenesis.

Pre-existing cardiovascular disease, acute kidney injury, and cardiovascular outcomes in hospitalized blacks with COVID-19 infection

BACKGROUND: The Corona Virus 19 (COVID-19) infection is associated with worse outcomes in blacks, although the mechanisms are unclear. We sought to determine the significance of black race, pre-existing cardiovascular disease (pCVD), and acute kidney injury (AKI) on cardiopulmonary outcomes and in-hospital mortality of COVID-19 patients. METHODS: We conducted a retrospective cohort study of blacks with/without pCVD and with/without in-hospital AKI, hospitalized within Grady Memorial Hospital in Georgia between February and July 2020, who tested positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on qualitative polymerase-chain-reaction assay. The primary outcome was a composite of in-hospital cardiac events. RESULTS: Of the 293 patients hospitalized with COVID-19 in this study, 71 were excluded from the primary analysis (for race/ethnicity other than black non-Hispanic). Of the 222 hospitalized COVID-19 patients included in our analyses, 41.4% were female, 78.8% had pCVD, and 30.6% developed AKI during the admission. In multivariable analyses, pCVD (OR 4.7, 95% CI 1.5-14.8, P=0.008) and AKI (OR 2.7, 95% CI 1.3-5.5, P=0.006) were associated with increased odds of in-hospital cardiac events. AKI was associated with increased odds of in-hospital mortality (OR 8.9, 95% CI 3.3-23.9, P<0.0001). The presence of AKI was associated with increased odds of ICU stay, mechanical ventilation, and acute respiratory distress syndrome (ARDS). CONCLUSION: pCVD and AKI were associated with higher risk of in-hospital cardiac events, and AKI was associated with a higher risk of in-hospital mortality in blacks.

Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell

Background: Established in vitro models for SARS-CoV-2 infection are limited and include cell lines of non-human origin and those engineered to overexpress ACE2, the cognate host cell receptor. Although Calu-3, a human lung cell line which endogenously expresses ACE2, supports SARS-CoV-2 replication, they are significantly less permissive to infection than other models. Furthermore, ACE2 expression in the respiratory tract is low and emerging evidence suggests the utilization of alternative host cell receptors and attachment factors may compensate for low ACE2 expression levels in the lung. We identified human H522 lung adenocarcinoma cells as naturally permissive to SARS-CoV-2 infection despite complete absence of ACE2. Methods: A panel of 10 cell lines, with variable expression levels of ACE2 and TMPRSS2 were infected with SARS-CoV-2 strain 2019-nCoV/USA-WA1/2020. Viral replication was monitored through assessment of cell-associated and cell-free viral RNA (vRNA) by QRT-PCR as well as N staining by FACS and in situ hybridization. Effect of blocking S protein by neutralizing antibodies and an ACE2-Fc decoy peptide, ACE2 blocking by a specific antibody, and ACE2 knockout by CRISPR on SARS-CoV-2 replication was determined by Q-RT-PCR for vRNAs. Various viral entry inhibitors were used to pathway of SARS-CoV-2 entry in H522 cells. RNA sequencing and proteomics was used to study the cell and innate immune responses in infected H522 cells. siRNA-mediated knockdown was utilized to further characterize the pathway of immune sensing. Results: Infection of H522 cells required the SARS-CoV-2 spike protein, though in contrast to ACE2-dependent models, spike alone was not sufficient for H522 infection. Temporally resolved transcriptomic and proteomic profiling revealed alterations in cell cycle and the antiviral host cell response, including MDA5- dependent activation of type-I interferon signaling. Focused chemical screens point to important roles for clathrin-mediated endocytosis and endosomal cathepsins in SARS-CoV-2 infection of H522 cells. Conclusion: These findings imply the utilization of an alternative SARS-CoV-2 host cell receptor which may impact tropism of SARS-CoV-2 and consequently human disease pathogenesis.