Development and characterization of a Yucatan miniature biomedical pig permanent middle cerebral artery occlusion stroke model

BACKGROUND: Efforts to develop stroke treatments have met with limited success despite an intense need to produce novel treatments. The failed translation of many of these therapies in clinical trials has lead to a close examination of the therapeutic development process. One of the major factors believed to be limiting effective screening of these treatments is the absence of an animal model more predictive of human responses to treatments. The pig may potentially fill this gap with a gyrencephalic brain that is larger in size with a more similar gray-white matter composition to humans than traditional stroke animal models. In this study we develop and characterize a novel pig middle cerebral artery occlusion (MCAO) ischemic stroke model. METHODS: Eleven male pigs underwent MCAO surgery with the first 4 landrace pigs utilized to optimize stroke procedure and 7 additional Yucatan stroked pigs studied over a 90 day period. MRI analysis was done at 24 hrs and 90 days and included T2w, T2w FLAIR, T1w FLAIR and DWI sequences and associated ADC maps. Pigs were sacrificed at 90 days and underwent gross and microscopic histological evaluation. Significance in quantitative changes was determined by two-way analysis of variance and post-hoc Tukey’s Pair-Wise comparisons. RESULTS: MRI analysis of animals that underwent MCAO surgery at 24 hrs had hyperintense regions in T2w and DWI images with corresponding ADC maps having hypointense regions indicating cytotoxic edema consistent with an ischemic stroke. At 90 days, region of interest analysis of T1 FLAIR and ADC maps had an average lesion size of 59.17 cc, a loss of 8% brain matter. Histological examination of pig brains showed atrophy and loss of tissue, consistent with MRI, as well as glial scar formation and macrophage infiltration. CONCLUSIONS: The MCAO procedure led to significant and consistent strokes with high survivability. These results suggest that the pig model is potentially a robust system for the study of stroke pathophysiology and potential diagnostics and therapeutics

Stability and Change of Neuroticism in Aging

Data from the Longitudinal Aging Study Amsterdam were used to study the relationship between neuroticism and aging. At baseline, cross-sectional analyses of data from 2,117 respondents (aged 55–85 years, M = 70) showed no significant age differences. The magnitude of the 3- and 6-year stability coefficients was high, and 12% of the elderly participants showed a clinically relevant mean level change. Longitudinal multilevel analyses showed a small but statistical significant change with aging, but the mean change was not considered clinically relevant. A U-formed course was found, showing a slight decrease until respondents reached the age of 70. Adjusting the model for physical health-related variables slightly increased the stability. An additional interaction analysis showed that the individual trajectory of neuroticism was not affected by the physical health status. In conclusion, neuroticism remains rather stable in middle and older adulthood, with some apparent increase in late life

Evaluation and calibration of Aeroqual Series 500 portable gas sensors for accurate measurement of ambient ozone and nitrogen dioxide

Low-power, and relatively low-cost, gas sensors have potential to improve understanding of intra-urban air pollution variation by enabling data capture over wider networks than is possible with 'traditional' reference analysers. We evaluated an Aeroqual Ltd. Series 500 semiconducting metal oxide O3 and an electrochemical NO2 sensor against UK national network reference analysers for more than 2months at an urban background site in central Edinburgh. Hourly-average Aeroqual O3 sensor observations were highly correlated (R2=0.91) and of similar magnitude to observations from the UV-absorption reference O3 analyser. The Aeroqual NO2 sensor observations correlated poorly with the reference chemiluminescence NO2 analyser (R2=0.02), but the deviations between Aeroqual and reference analyser values ([NO2]Aeroq-[NO2]ref) were highly significantly correlated with concurrent Aeroqual O3 sensor observations [O3]Aeroq. This permitted effective linear calibration of the [NO2]Aeroq data, evaluated using 'hold out' subsets of the data (R2≥0.85). These field observations under temperate environmental conditions suggest that the Aeroqual Series 500 NO2 and O3 monitors have good potential to be useful ambient air monitoring instruments in urban environments provided that the O3 and NO2 gas sensors are calibrated against reference analysers and deployed in parallel

Quality of Life of Caregivers of Older Patients with Advanced Cancer

OBJECTIVES: To evaluate the relationships between aging-related domains captured by geriatric assessment (GA) for older patients with advanced cancer and caregivers’ emotional health and quality of life (QOL). DESIGN: In this cross sectional study of baseline data from a nationwide investigation of older patients and their caregivers, patients completed a GA that included validated tests to evaluate eight domains of health (eg, function, cognition). SETTING: Thirty-one community oncology practices throughout the United States. PARTICIPANTS: Enrolled patients were aged 70 and older, had one or more GA domain impaired, and had an incurable solid tumor malignancy or lymphoma. Each could choose one caregiver to enroll. MEASUREMENTS: Caregivers completed the Generalized Anxiety Disorder-7, Distress Thermometer, Patient Health Questionnaire-2 (depression), and Short Form Health Survey-12 (SF-12 for QOL). Separate multivariate linear or logistic regression models were used to examine the association of the number and type of patient GA impairments with caregiver outcomes, controlling for patient and caregiver covariates. RESULTS: A total of 541 patients were enrolled, 414 with a caregiver. Almost half (43.5%) of the caregivers screened positive for distress, 24.4% for anxiety, and 18.9% for depression. Higher numbers of patient GA domain impairments were associated with caregiver depression (adjusted odds ratio [aOR] = 1.29; P <.001], caregiver physical health on SF-12 (regression coefficient [β] = −1.24; P <.001), and overall caregiver QOL (β = −1.14; P <.01). Impaired patient function was associated with lower caregiver QOL (β = −4.11; P <.001). Impaired patient nutrition was associated with caregiver depression (aOR = 2.08; P <.01). Lower caregiver age, caregiver comorbidity, and patient distress were also associated with worse caregiver outcomes. CONCLUSION: Patient GA impairments were associated with poorer emotional health and lower QOL of caregivers

Open science discovery of potent noncovalent SARS-CoV-2 main protease inhibitors

INTRODUCTION COVID-19 became a global pandemic partially as a result of the lack of easily deployable, broad-spectrum oral antivirals, which complicated its containment. Even endemically, and with effective vaccinations, it will continue to cause acute disease, death, and long-term sequelae globally unless there are accessible treatments. COVID-19 is not an isolated event but instead is the latest example of a viral pandemic threat to human health. Therefore, antiviral discovery and development should be a key pillar of pandemic preparedness efforts. RATIONALE One route to accelerate antiviral drug discovery is the establishment of open knowledge bases, the development of effective technology infrastructures, and the discovery of multiple potent antivirals suitable as starting points for the development of therapeutics. In this work, we report the results of the COVID Moonshot—a fully open science, crowdsourced, and structure-enabled drug discovery campaign—against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro). This collaboration may serve as a roadmap for the potential development of future antivirals. RESULTS On the basis of the results of a crystallographic fragment screen, we crowdsourced design ideas to progress from fragment to lead compounds. The crowdsourcing strategy yielded several key compounds along the optimization trajectory, including the starting compound of what became the primary lead series. Three additional chemically distinct lead series were also explored, spanning a diversity of chemotypes. The collaborative and highly automated nature of the COVID Moonshot Consortium resulted in >18,000 compound designs, >2400 synthesized compounds, >490 ligand-bound x-ray structures, >22,000 alchemical free-energy calculations, and >10,000 biochemical measurements—all of which were made publicly available in real time. The recently approved antiviral ensitrelvir was identified in part based on crystallographic data from the COVID Moonshot Consortium. This campaign led to the discovery of a potent [median inhibitory concentration (IC50) = 37 ± 2 nM] and differentiated (noncovalent and nonpeptidic) lead compound that also exhibited potent cellular activity, with a median effective concentration (EC50) of 64 nM in A549-ACE2-TMPRSS2 cells and 126 nM in HeLa-ACE2 cells without measurable cytotoxicity. Although the pharmacokinetics of the reported compound is not yet optimal for therapeutic development, it is a promising starting point for further antiviral discovery and development. CONCLUSION The success of the COVID Moonshot project in producing potent antivirals, building open knowledge bases, accelerating external discovery efforts, and functioning as a useful information-exchange hub is an example of the potential effectiveness of open science antiviral discovery programs. The open science, patent-free nature of the project enabled a large number of collaborators to provide in-kind support, including synthesis, assays, and in vitro and in vivo experiments. By making all data immediately available and ensuring that all compounds are purchasable from Enamine without the need for materials transfer agreements, we aim to accelerate research globally along parallel tracks. In the process, we generated a detailed map of the structural plasticity of Mpro, extensive structure-activity relationships for multiple chemotypes, and a wealth of biochemical activity data to spur further research into antivirals and discovery methodologies. We hope that this can serve as an alternative model for antiviral discovery and future pandemic preparedness. Further, the project also showcases the role of machine learning, computational chemistry, and high-throughput structural biology as force multipliers in drug design. Artificial intelligence and machine learning algorithms help accelerate chemical synthesis while balancing multiple competing molecular properties. The design-make-test-analyze cycle was accelerated by these algorithms combined with planetary-scale biomolecular simulations of protein-ligand interactions and rapid structure determination

SARS-CoV-2 infects the human kidney and drives fibrosis in kidney organoids

Kidney failure is frequently observed during and after COVID-19, but it remains elusive whether this is a direct effect of the virus. Here, we report that SARS-CoV-2 directly infects kidney cells and is associated with increased tubule-interstitial kidney fibrosis in patient autopsy samples. To study direct effects of the virus on the kidney independent of systemic effects of COVID-19, we infected human-induced pluripotent stem-cell-derived kidney organoids with SARS-CoV-2. Single-cell RNA sequencing indicated injury and dedifferentiation of infected cells with activation of profibrotic signaling pathways. Importantly, SARS-CoV-2 infection also led to increased collagen 1 protein expression in organoids. A SARS-CoV-2 protease inhibitor was able to ameliorate the infection of kidney cells by SARS-CoV-2. Our results suggest that SARS-CoV-2 can directly infect kidney cells and induce cell injury with subsequent fibrosis. These data could explain both acute kidney injury in COVID-19 patients and the development of chronic kidney disease in long COVID

Watershed and Estuarine Controls Both Influence Plant Community and Tree Growth Changes in Tidal Freshwater Forested Wetlands along Two U.S. Mid-Atlantic Rivers

The tidal freshwater zone near the estuarine head-of-tide is potentially sensitive to both sea-level rise and associated salinity increases as well as changing watershed inputs of freshwater and nutrients. We evaluated the vegetation response of tidal freshwater forested wetlands (TFFW) to changes in nontidal river versus estuarine controls along the longitudinal gradient of the Mattaponi and Pamunkey rivers in the Mid-Atlantic USA. The gradient included nontidal freshwater floodplain (NT) and upper tidal (UT), lower tidal (LT), and stressed tidal forest transitioning to marsh (ST) TFFW habitats on both rivers. Plot-based vegetation sampling and dendrochronology were employed to examine: (1) downriver shifts in plant community composition and the structure of canopy trees, understory trees/saplings/shrubs and herbs, tree basal-area increment (BAI) and (2) interannual variability in BAI from 2015 dating back as far as 1969 in relation to long-term river and estuary monitoring data. With greater tidal influence downstream, tree species dominance shifted, live basal area generally decreased, long-term mean BAI of individual trees decreased, woody stem mortality increased, and live herbaceous vegetative cover and richness increased. Acer rubrum, Fagus grandifolia, Ilex opaca, and Fraxinus pennsylvanica dominated NT and UT sites, with F. pennsylvanica and Nyssa sylvatica increasingly dominating at more downstream tidal sites. Annual tree BAI growth was positively affected by nontidal river flow at NT and UT sites which were closer to the head-of-tide, positively influenced by small salinity increases at LT and ST sites further downstream, and positively influenced by estuarine water level throughout the gradient; nutrient influence was site specific with both positive and negative influences. The counterintuitive finding of salinity increasing tree growth at sites with low BAI is likely due to either competitive growth release from neighboring tree death or enhanced soil nutrient availability that may temporarily mitigate the negative effects of low-level salinization and sea-level increases on living TFFW canopy trees, even as overall plant community conversion to tidal marsh progresses