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Mutations in 5’ untranslated regions (UTRs) have a functional role in gene expression in cancer. Here, the authors develop a sequencing-based high throughput functional assay named PLUMAGE and show the effects of these mutations on gene expression and their association with clinical outcomes in prostate cancer.

This meta-analysis investigated the association between age-related hearing loss and structural neuroanatomy, specifically changes to gray matter volume. Hearing loss is associated with increased risk of cognitive decline. Hence, understanding the effects of hearing loss in older age on brain health is essential. We reviewed studies which compared older participants with hearing loss (age-related hearing loss: ARHL) to older adults without clinical hearing loss (no-ARHL), on neuroanatomical outcomes, specifically gray matter (GM) volume as measured by magnetic resonance imaging. A total of five studies met the inclusion criteria, three of which were included in an analysis of whole-brain gray matter volume (ARHL group n = 113; no-ARHL group n = 138), and three were included in analyses of lobe-wise gray matter volume (ARHL group n = 139; no-ARHL group n = 162). Effect-size seed-based d mapping software was employed for whole-brain and lobe-wise analysis of gray matter volume. The analysis indicated there was no significant difference between adults with ARHL compared to those with no-ARHL in whole-brain gray matter volume. Due to lacking stereotactic coordinates, the level of gray matter in specific neuroanatomical locations could only be observed at lobe-level. These data indicate that adults with ARHL show increased gray matter atrophy in the temporal lobe only (not in occipital, parietal, or frontal), compared to adults with no-ARHL. The implications for theoretical frameworks of the hearing loss and cognitive decline relationship are discussed in relation to the results. This meta-analysis was pre-registered on PROSPERO (CRD42021265375).Systematic Review Registration:https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=265375, PROSPERO CRD42021265375.

Abstract Objectives The use of online imagery by non-local observers to conduct remote, centralized collection of streetscape audit data in international studies has the potential to enhance efficiency of collection and comparability of such data for research on built environments and health. The objectives of the study were to measure (1) the consistency in responses between local in-field observers and non-local remote online observers and (2) the reliability between in-country online observers and non-local remote online observers using the Microscale Audit of Pedestrian Streetscapes Global tool to characterize pedestrian-related features along streets in five countries. Methods Consistency and inter-rater reliability were analyzed between local and non-local observers on a pooled database of 200 routes in five study regions (Melbourne, Australia; Ghent, Belgium; Curitiba, Brazil; Hong Kong, China; and Valencia, Spain) for microscale environmental feature subscales and item-level variables using the intraclass correlation coefficient (ICC). Results A local in-field versus remote online comparison had an ICC of 0.75 (95 % CI: 0.68–0.80) for the grand total score. An ICC of 0.91 (95 % CI: 0.88–0.93) was found for the local online versus remote online comparison. Positive subscales yielded stronger results in comparison to negative subscales, except for the similarly poor-performing positive aesthetics/social characteristics. Conclusions This study demonstrated remote audits of microscale built environments using online imagery had good reliability with local in-field audits and excellent reliability with local online audits. Results generally supported remote online environmental audits as comparable to local online audits. This identification of low-cost and efficient data acquisition methods is important for expanding research on microscale built environments and physical activity globally.

Mounting evidence shows that pollinators are declining as a result of widespread environmental degradation. This loss raises concerns that a global pollination crisis could threaten the human food supply by decreasing crop yield and even promote famine under a hypothetical scenario of total pollinator extinction. This catastrophic possibility has prompted intense interest from scientists, politicians and the general public. However, three lines of evidence do not support such an apocalyptic scenario. First, even though the abundance and diversity of wild pollinators are declining worldwide, the global population of managed honey-bee hives has increased by ~80% since the early 1960s. Second, agricultural production would decrease by

BackgroundJust-in-time adaptive interventions (JITAIs) provide real time in-the-moment behavior change support to people when they need it most. JITAIs could be a viable way to provide personalized physical activity (PA) support to older adults in the community. However, it is unclear how feasible it is to remotely deliver a PA intervention through a smartphone to older adults or how acceptable they would find a JITAI targeting PA in everyday life. ObjectiveThe aims of this study are to describe the development of JitaBug, a personalized smartphone-delivered JITAI designed to support older adults to increase or maintain their PA level, assess the feasibility of conducting an effectiveness trial of the JitaBug intervention, and explore the acceptability of JitaBug among older adults in a free-living setting. MethodsThe intervention was developed using the Behavior Change Wheel and consisted of a wearable activity tracker (Fitbit) and a companion smartphone app (JitaBug) that delivered goal-setting, planning, reminders, and JITAI messages to encourage achievement of personalized PA goals. Message delivery was tailored based on time of day, real time PA tracker data, and weather conditions. We tested the feasibility of remotely delivering the intervention with older adults in a 6-week trial. Data collection involved assessment of PA through accelerometery and activity tracker, self-reported mood and mental well-being through ecological momentary assessment, and contextual information on PA through voice memos. Feasibility outcomes included recruitment capability and adherence to the intervention, intervention delivery in the wild, appropriateness of data collection methodology, adverse events, and participant satisfaction. ResultsOf the 46 recruited older adults (aged 56-72 years), 31 (67%) completed the intervention. The intervention was successfully delivered as intended; 87% (27/31) of the participants completed the intervention independently; 94% (2247/2390) of the PA messages were successfully delivered; 99% (2239/2261) of the Fitbit and 100% (2261/2261) of the weather data calls were successful. Valid and usable wrist-worn accelerometer data were obtained from 90% (28/31) of the participants at baseline and follow-up. On average, the participants recorded 50% (7.9/16, SD 7.3) of the voice memos, 38% (3.3/8, SD 4.2) of the mood assessments, and 50% (2.1/4, SD 1.6) of the well-being assessments through the app. Overall acceptability of the intervention was very good (23/30, 77% expressed satisfaction). Participant feedback suggested that more diverse and tailored PA messages, app use reminders, technical refinements, and an improved user interface could improve the intervention and make it more appealing. ConclusionsThis study suggests that a smartphone-delivered JITAI is an acceptable way to support PA in older adults in the community. Overall, the intervention is feasible; however, based on user feedback, the JitaBug app requires further technical refinements that may enhance use, engagement, and user satisfaction before moving to effectiveness trials.

The inter- and intra-tumor heterogeneity of metastatic prostate cancer (mPC) is underexplored. Here the authors use Digital Spatial Profiling to study gene and protein expression heterogeneity in 27 mPC patients, finding variation in associated pathways and potential immunotherapy targets.

The COVID-19 pandemic has generated increased interest in potential transmission routes. In food retail settings, transmission from infected customers and workers and customers through surfaces has been deemed plausible. However, limited information exists on the presence and survival of SARS-CoV-2 on surfaces, particularly outside laboratory settings. Therefore, the purpose of this project was to assess the presence of the virus at commonly found surfaces at food retail stores and the potential role that these spaces play in virus transmission.Samples (n=957) were collected twice a week for a month in food-retail stores within Ontario, Canada. High-touch surfaces were identified and surveyed in 4 zones within the store (payment stations, deli counters, refrigerated food section and carts and baskets). The samples were analyzed using a molecular method, i.e., reverse transcriptase quantitative Polymerase Chain Reaction (RT-qPCR).Regardless of the store's location, the sampling day or time, the location of the surface within the store or the surface material, all samples tested negative for SARS-CoV-2. These results suggest that the risk of exposure from contaminated high-touch surfaces within a food retailer store is low if preventive measures and recommended sanitizing routines are maintained.

Lead acetate associated tissue injury has been linked to altered antioxidant defenses, hyperuricemia and inflammation. We hypothesized that watermelon rind extract, would ameliorate lead acetate-induced hepato-renal injury.Thirty Male Wistar rats received distilled water, lead acetate (Pb; 5 mg/kg) with or without watermelon rind extract (WM; 400 mg/kg; WM + Pb; 15 days of WM pretreatment); Pb + WM (15 days of WM post treatment) and simultaneous treatment (WM-Pb) for 30 days.Lead toxicity led to elevated serum malondialdehyde, creatinine, urea, uric acid, lactate dehydrogenase, liver injury enzymes, as well as decreased body weight. Decreased serum levels of reduced glutathione, nitric oxide, total protein and glutathione peroxidase activity was also observed. However, these alterations were ameliorated by watermelon rind extract in lead acetate-treated rats.Watermelon rind ethanol extract protects against lead acetate-induced hepato-renal injury through improved antioxidant defenses at least in part, via uric acid/nitric oxide-dependent pathway signifying the health benefits of this agricultural waste and a potential for waste recycling while limiting environmental pollution.

The presence of missing values in a time-series dataset is a very common and well-known problem. Various statistical and machine learning methods have been developed to overcome this problem, with the aim of filling in the missing values in the data. However, the performances of these methods vary widely, showing a high dependence on the type of data and correlations within the data. In our study, we performed some of the well-known imputation methods, such as expectation maximization, k-nearest neighbor, iterative imputer, random forest, and simple imputer, to impute missing data obtained from smart, wearable health trackers. In this manuscript, we proposed the use of data binning for imputation. We showed that the use of data binned around the missing time interval provides a better imputation than the use of a whole dataset. Imputation was performed for 15 min and 1 h of continuous missing data. We used a dataset with different bin sizes, such as 15 min, 30 min, 45 min, and 1 h, and we carried out evaluations using root mean square error (RMSE) values. We observed that the expectation maximization algorithm worked best for the use of binned data. This was followed by the simple imputer, iterative imputer, and k-nearest neighbor, whereas the random forest method had no effect on data binning during imputation. Moreover, the smallest bin sizes of 15 min and 1 h were observed to provide the lowest RMSE values for the majority of the time frames during the imputation of 15 min and 1 h of missing data, respectively. Although applicable to digital health data, we think that this method will also find applicability in other domains.

Around 5.5–6.5 months of age, infants first attend to object size and perceive its mass cues in simple collision events. Infants attend to the size of the moving object and expect a greater displacement following a collision with a large object and stationary object, and lesser displacement following a collision with a small object and stationary object. It has been proposed that infants of 6-to-7 months of age can differentiate between sizes of moving objects but do not perceive the size and mass relationships in simple collision events. The present two investigations aimed to investigate whether infants 10-to-11 months of age (N = 16) could perceive this relationship (experiment 1) and the reverse of this relationship (experiment 2) utilising the looking time paradigm. The reverse of this relationship entailed the circumstances in which the moving object size was kept constant, but the stationary object size varied (small or large). Results from these experiments revealed that infants did not differ in their looking times for size congruent and size incongruent distances in both conditions. Infants did not look longer at the incongruent test events that violated expectation. For that reason, we conclude infants of 10-to-11 months of age were unable to perceive size and mass associations in collision events in either direction (moving object or stationary object size).

This study examined the differences in the level of physical fitness and academic performance among male middle-school children based on different body status categories. A total of 69 male children [age: 12.4 ± 0.7 years; body mass: 58.5 ± 7.2 kg; height: 1.62 ± 0.09 m; and body mass index (BMI): 22.4 ± 3.3 kg/m2] participated and were divided into BMI age-adjusted groups (i.e., lowest, middle, and highest BMI). Height, mass, BMI, stork test of static balance, 10 and 15 m sprint as an indicator for speed, hand-grip strength test, agility T-half test, medicine ball throw (MBT), and the Yo-Yo Intermittent Recovery Test level 1 (Yo-Yo IR1) were assessed. School records were retrieved for grade point averages (GPA) of mathematics, science, and Arabic. We found significant group differences regarding anthropometric (height: ηp2 = 0.24, mass: ηp2 = 0.33, and BMI: ηp2 = 0.66), physical (sprint 10 m: ηp2 = 0.26), and academic (mathematics: ηp2 = 0.19 and science: ηp2 = 0.15) performance parameters. The largest difference (p < 0.001) was observed between the lowest and highest group for the 10 m sprint. All pairwise differences were between the lowest and highest BMI group or the lowest and middle BMI group. No relevant (r > 0.5) correlation between parameters of different dimensions (e.g., anthropometric vs. physical performance parameters) was found. In conclusion, the highest BMI group exhibited similar physical and academic performances than the lowest group. Thus, these data emphasize the importance and appropriateness to engage young Qatari schoolchildren in physical activity as it associates with superior academic performance.

The microbial quality of agricultural water is often assessed using fecal indicator bacteria (FIB) and physicochemical parameters. The presence, direction, and strength of associations between microbial and physicochemical parameters, and the presence of human pathogens in surface water vary across space (e.g., region) and time. This study was undertaken to understand these associations in two produce-growing regions in Florida, USA, and to examine the pathogen ecology in waterways used for produce production. The relationship between Salmonella presence, and microbial and physicochemical water quality; as well as weather and land use factors were evaluated. Water samples were collected from six sites in North Florida (N = 72 samples) and eight sites in South Florida (N = 96 samples) over 12 sampling months. Land use around each sampling site was characterized, and weather and water quality data were collected at each sampling. Salmonella, generic Escherichia coli, total coliform, and aerobic plate count bacteria populations were enumerated in each sample. Univariable and multivariable regression models were then developed to characterize associations between microbial water quality (i.e., E. coli levels and Salmonella presence), and water quality, weather, and land use factors separately for North and South Florida. The E. coli and total coliforms mean concentrations (log10 MPN/100 mL) were 1.8 ± 0.6 and >3.0 ± 0.4 in North and 1.3 ± 0.6 and >3.3 ± 0.2 in South Florida waterways, respectively. While Salmonella was detected in 23.6% (17/72) of North Florida and 28.1% (27/96) of South Florida samples, the concentration ranged between

This study examined the effect of 8 weeks of biweekly combined plyometric and short sprint training into the typical within-season training schedule of youth male soccer players. Participants were allocated at random to an experimental group (EG; n = 17, age: 14.6 ± 0.5 years, body mass: 60.5 ± 7.1 kg, height: 1.64 ± 0.08 m, body fat: 11.3 ± 1.4%) and a control group (CG; n = 17, age: 14.6 ± 0.4 years, body mass: 61.0 ± 3.9 kg, height: 1.67 ± 0.05 m, body fat: 11.8 ± 1.4%). Measures obtained pre- and post-intervention included vertical and horizontal jump performances (i.e., squat jump (SJ), countermovement jump with aimed arms (CMJA), and five-jump test (FJT)) and sprint performances (i.e., 10 and 30 m sprint). In addition, change-of-direction ability (sprint with 90° Turns (S90°) and sprint 9–3–6–3–9 m with backward and forward running (SBF)), repeated shuttle sprint ability (RSSA), and dynamic balance performance (Y balance test) were measured pre- and post-intervention. The EG experienced higher jump (all p < 0.05; d ≥ 0.71), sprint (all p < 0.05; d ≥ 0.64), change-of-direction ability (all p < 0.05; d ≥ 0.66), RSSA (all parameters except the fatigue index p < 0.01; d ≥ 0.71), and dynamic balance (all p ≤ 0.05; d ≥ 0.50) improvement compared to the CG. Adding biweekly combined plyometric and short sprint training to standard training improves the athletic performance of youth male soccer players (under 15 (U15)).

Background: Diabetes is among the most prevalent non-communicable diseases causing significant morbidity and mortality globally. The aetiology and disease development of diabetes are influenced by genetic, lifestyle, and environmental factors. Due to an increasing number of cases each year, it is imperative to improve the understanding of modifiable environmental risk and protective factors. In this study we aimed to analyse associations between built and natural environment features and diabetes prevalence; and two major risk factors: physical activity and obesity and their mediation effects. Methods: We analysed relationships between walkability and park availability with physical activity, obesity, and diabetes, using self-reported data from a large cross-sectional survey in British Columbia, Canada (n = 22,418). We validated results with an independent cohort (n = 11,972) in a subset of the analyses. The outcome measures included walking, moderate to vigorous physical activity (MVPA), body mass index (BMI), and diabetes. Built and natural environment features within a 1 km road network buffer around residential postal code centroids were assessed using validated indicators of walkability and park availability. We used general linear multivariable models (GLM) to examine the direct relationship between environmental features, physical activity, obesity, and diabetes respectively. Path models were developed to analyse mediation effects of physical activity and obesity on the association between environmental indicators and diabetes. The relative contribution of direct versus indirect effects was assessed. All models were adjusted for age, gender, income. ethnicity, years lived in neighbourhood and regional accessibility. Results: Walkable neighbourhoods and areas with greater park availability were associated with lower rates of diabetes. There was a direct association of walkability and park availability on physical activity (highest vs. lowest quintile OR = 1.15; 95% CI: 1.00, 1.33 and OR = 1.28, 95% CI: 1.13, 1.45 respectively), obesity (highest vs. lowest quintile OR = 0.58, 95% CI: 0.49, 0.70 and OR = 0.57, 95% CI: 0.48, 0.68 respectively) and diabetes (highest vs. lowest quintile OR = 0.62, 95% CI: 0.45, 0.85, and OR = 0.63, 95% CI: 0.47, 0.84 respectively). Results were similar in the independent cohort. The associations between diabetes and walkability and park availability respectively were partly mediated by obesity (41% of total association for walkability and 53% of total association for park availability). The mediating effect of physical activity was negligible. Conclusion: Results support investments in walkability through active transportation and transit infrastructure. Changes in zoning and subdivision regulations governing land use actions are required to enable compact mixed-use environments with access to parks and high quality transit service. Future studies including cost-benefit analyses of health-related economic impacts of such investments can contribute to evidence-based decisions for healthier cities.

The rapid global rise of COVID-19 from late 2019 caught major manufacturers of RT-qPCR reagents by surprise and threw into sharp focus the heavy reliance of molecular diagnostic providers on a handful of reagent suppliers. In addition, lockdown and transport bans, necessarily imposed to contain disease spread, put pressure on global supply lines with freight volumes severely restricted. These issues were acutely felt in New Zealand, an island nation located at the end of most supply lines. This led New Zealand scientists to pose the hypothetical question: in a doomsday scenario where access to COVID-19 RT-qPCR reagents became unavailable, would New Zealand possess the expertise and infrastructure to make its own reagents onshore? In this work we describe a review of New Zealand's COVID-19 test requirements, bring together local experts and resources to make all reagents for the RT-qPCR process, and create a COVID-19 diagnostic assay referred to as HomeBrew (HB) RT-qPCR from onshore synthesized components. This one-step RT-qPCR assay was evaluated using clinical samples and shown to be comparable to a commercial COVID-19 assay. Through this work we show New Zealand has both the expertise and, with sufficient lead time and forward planning, infrastructure capacity to meet reagent supply challenges if they were ever to emerge.

Abstract Background Investigation of physical activity and dietary behaviors across locations can inform “setting-specific” health behavior interventions and improve understanding of contextual vulnerabilities to poor health. This study examined how physical activity, sedentary time, and dietary behaviors differed across home, school, and other locations in young adolescents. Methods Participants were adolescents aged 12–16 years from the Baltimore-Washington, DC and the Seattle areas from a larger cross-sectional study. Participants (n = 472) wore an accelerometer and Global Positioning Systems (GPS) tracker (Mean days = 5.12, SD = 1.62) to collect location-based physical activity and sedentary data. Participants (n = 789) completed 24-h dietary recalls to assess dietary behaviors and eating locations. Spatial analyses were performed to classify daily physical activity, sedentary time patterns, and dietary behaviors by location, categorized as home, school, and “other” locations. Results Adolescents were least physically active at home (2.5 min/hour of wear time) and school (2.9 min/hour of wear time) compared to “other” locations (5.9 min/hour of wear time). Participants spent a slightly greater proportion of wear time in sedentary time when at school (41 min/hour of wear time) than at home (39 min/hour of wear time), and time in bouts lasting ≥30 min (10 min/hour of wear time) and mean sedentary bout duration (5 min) were highest at school. About 61% of daily energy intake occurred at home, 25% at school, and 14% at “other” locations. Proportionately to energy intake, daily added sugar intake (5 g/100 kcal), fruits and vegetables (0.16 servings/100 kcal), high calorie beverages (0.09 beverages/100 kcal), whole grains (0.04 servings/100 kcal), grams of fiber (0.65 g/100 kcal), and calories of fat (33 kcal/100 kcal) and saturated fat (12 kcal/100 kcal) consumed were nutritionally least favorable at “other” locations. Daily sweet and savory snacks consumed was highest at school (0.14 snacks/100 kcal). Conclusions Adolescents’ health behaviors differed based on the location/environment they were in. Although dietary behaviors were generally more favorable in the home and school locations, physical activity was generally low and sedentary time was higher in these locations. Health behavior interventions that address the multiple locations in which adolescents spend time and use location-specific behavior change strategies should be explored to optimize health behaviors in each location.

Here, Li et al. perform a genome-wide CRISPR screen to identify host dependency factors for influenza A virus infection and show that the host mRNA cap methyltransferase CMTR1 is important for viral cap snatching and that it affects expression of antiviral genes.

Abstract Background Parkinson’s disease (PD) is characterised by dopaminergic cell loss within the substantia nigra pars compacta (SNc) that leads to reduced striatal dopamine content and resulting motor deficits. Identifying new strategies to protect these cells from degeneration and retain striatal dopaminergic innervation is therefore of great importance. Chondroitin sulphate proteoglycans (CSPGs) are recognised contributors to the inhibitory extracellular milieu known to hinder tissue recovery following CNS damage. Digestion of these molecules by the bacterial lyase chondroitinase ABC (ChABC) has been shown to promote functional recovery in animal models of neurological injury. Although ChABC has been shown to promote sprouting of dopaminergic axons following transection of the nigrostriatal pathway, its ability to protect against nigrostriatal degeneration in a toxin-based module with better construct validity for PD has yet to be explored. Here we examined the neuroprotective efficacy of ChABC treatment in the full and partial 6-hydroxydopamine (6-OHDA) lesion mouse models of PD. Results In mice bearing a full 6-OHDA lesion, ChABC treatment failed to protect against the loss of either nigral cells or striatal terminals. In contrast, in mice bearing a partial 6-OHDA lesion, ChABC treatment significantly protected cells of the rostral SNc, which remained at more than double the numbers seen in vehicle-treated animals. In the partial lesion model, ChABC treatment also significantly preserved dopaminergic fibres of the rostral dorsal striatum which increased from 15.3 ± 3.5% of the intact hemisphere in saline-treated animals to 36.3 ± 6.5% in the ChABC-treated group. These protective effects of ChABC treatment were not accompanied by improvements in either the cylinder or amphetamine-induced rotations tests of motor function. Conclusions ChABC treatment provided significant protection against a partial 6-OHDA lesion of the nigrostriatal tract although the degree of protection was not sufficient to improve motor outcomes. These results support further investigations into the benefits of ChABC treatment for providing neuroprotection in PD.

Stimulated Raman scattering (SRS) microscopy enables label-free chemical imaging at high speed, but has been limited by low sensitivity. Here, the authors demonstrate plasmon-enhanced SRS microscopy and achieve single molecule detection sensitivity.

Matthew B Schabath,1,2 Tapashi B Dalvi,3 Hongyue A Dai,4 Alan L Crim,4 Anita Midha,5 Norah Shire,3 Nicholas T Gimbrone,1 Jill Walker,6 Danielle M Greenawalt,7 David Lawrence,8 James R Rigas,9 Robert Brody,9 Danielle Potter,9 Naveen S Kumar,4 Shane A Huntsman,4 Jhanelle E Gray2 1Department of Cancer Epidemiology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA; 2Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA; 3Oncology R&D, AstraZeneca, Gaithersburg, MD, USA; 4M2Gen, Tampa, FL, USA; 5Department of Personalised Healthcare and Biomarkers, AstraZeneca, Cambridge, UK; 6Department of Precision Medicine Oncology, AstraZeneca, Cambridge, UK; 7Department of iMED Oncology Informatics, AstraZeneca, Waltham, MA, USA; 8Department of Global Medicines Development, AstraZeneca, Cambridge, UK; 9Department of Global Medical Affairs Oncology, AstraZeneca, Gaithersburg, MD, USACorrespondence: Matthew B SchabathH. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USATel +1 813 745 4150Fax +1 813 745 6525Email matthew.schabath@moffitt.orgPurpose: To characterize programmed cell death ligand-1 (PD-L1) expression in relation to survival and gene mutation status in patients with advanced NSCLC. The study also explored the influence of tumor mutational burden (TMB) on PD-L1 expression and patient characteristics.Patients and methods: Adult patients with histologically or cytologically documented Stage IIIB/Stage IV/recurrent/progressive NSCLC, Eastern Cooperative Oncology Group performance status 0 to 3, and >2 lines of prior systemic treatment regimens were included in this retrospective analysis. Patients were treated from 1997 to 2015 at H. Lee Moffitt Cancer Center and Research Institute, Tampa, or at 7 community centers across the United States. PD-L1 expression level was determined using the VENTANA PD-L1 (SP263) Assay. EGFR and KRAS mutation status and ALK rearrangements were determined by targeted DNA sequencing; these were obtained from clinical records where targeted DNA sequencing was not performed. TMB was calculated as the total number of somatic mutations per sample.Results: From a total of 136 patients included in the study, 23.5% had tumors with high PD-L1 expression (≥25%). There were no significant differences in patient characteristics, overall survival (OS), and progression-free survival (PFS) between patients with high PD-L1 expression (median OS: 39.5 months; median PFS: 15.8 months) vs low PD-L1 expression (

Epibionts are organisms that utilize the exterior of other organisms as a living substratum. Many affiliate opportunistically with hosts of different species, but others specialize on particular hosts as obligate associates. We investigated a case of apparent host specificity between two barnacles that are epizoites of sea turtles and illuminate some ecological considerations that may shape their host relationships. The barnacles Chelonibia testudinaria and Chelonibia caretta, though roughly similar in appearance, are separable by distinctions in morphology, genotype, and lifestyle. However, though each is known to colonize both green (Chelonia mydas) and hawksbill (Eretmochelys imbricata) sea turtles, C. testudinaria is >5 times more common on greens, while C. caretta is >300 times more common on hawksbills. Two competing explanations for this asymmetry in barnacle incidence are either that the species’ larvae are spatially segregated in mutually exclusive host-encounter zones or their distributions overlap and the larvae behaviorally select their hosts from a common pool. We indirectly tested the latter by documenting the occurrence of adults of both barnacle species in two locations (SE Florida and Nose Be, Madagascar) where both turtle species co-mingle. For green and hawksbill turtles in both locations (Florida: n = 32 and n = 275, respectively; Madagascar: n = 32 and n = 125, respectively), we found that C. testudinaria occurred on green turtles only (percent occurrence – FL: 38.1%; MD: 6.3%), whereas the barnacle C. caretta was exclusively found on hawksbill turtles (FL: 82.2%; MD: 27.5%). These results support the hypothesis that the larvae of these barnacles differentially select host species from a shared supply. Physio-biochemical differences in host shell material, conspecific chemical cues, external microbial biofilms, and other surface signals may be salient factors in larval selectivity. Alternatively, barnacle presence may vary by host micro-environment. Dissimilarities in scute structure and shell growth between hawksbill and green turtles may promote critical differences in attachment modes observed between these barnacles. In understanding the co-evolution of barnacles and hosts it is key to consider the ecologies of both hosts and epibionts in interpreting associations of chance, choice, and dependence. Further studies are necessary to investigate the population status and settlement spectrum of barnacles inhabiting sea turtles.

Background: Persistent coronavirus disease 2019 (COVID-19) symptoms are increasingly well-reported in cohort studies and case series. Given the spread of the pandemic, number of individuals suffering from persistent symptoms, termed ‘long COVID', are significant. However, type and prevalence of symptoms are not well reported using systematic literature reviews.Objectives: In this scoping review of the literature, we aggregated type and prevalence of symptoms in people with long COVID.Eligibility Criteria: Original investigations concerning the name and prevalence of symptoms were considered in participants ≥4-weeks post-infection.Sources of Evidence: Four electronic databases [Medline, Web of Science, Scopus, and the Cochrane Central Register of Controlled Trials (CENTRAL)] were searched.Methods: A scoping review was conducted using the Arksey and O'Malley framework. Review selection and characterisation was performed by three independent reviewers using pretested forms.Results: Authors reviewed 2,711 titles and abstracts for inclusion with 152 selected for full-text review. 102 articles were subsequently removed as this did not meet inclusion criteria. Thus, fifty studies were analysed, 34 of which were described as cohort studies or prospective cohort studies, 14 were described as cross-sectional studies, one was described as a case control study, and one was described as a retrospective observational study. In total, >100 symptoms were identified and there was considerable heterogeneity in symptom prevalence and setting of study. Ten studies reported cardiovascular symptoms, four examined pulmonary symptoms, 25 reported respiratory symptoms, 24 reported pain-related symptoms, 21 reported fatigue, 16 reported general infection symptoms, 10 reported symptoms of psychological disorders, nine reported cognitive impairment, 31 reported a sensory impairment, seven reported a dermatological complaint, 11 reported a functional impairment, and 18 reported a symptom which did not fit into any of the above categories.Conclusion: Most studies report symptoms analogous to those apparent in acute COVID-19 infection (i.e., sensory impairment and respiratory symptoms). Yet, our data suggest a larger spectrum of symptoms, evidenced by >100 reported symptoms. Symptom prevalence varied significantly and was not explained by data collection approaches, study design or other methodological approaches, and may be related to unknown cohort-specific factors.

Summary: Extracellular agonists linked to inositol-1,4,5-trisphosphate (IP3) formation elicit cytosolic Ca2+ oscillations in many cell types, but despite a common signaling pathway, distinct agonist-specific Ca2+ spike patterns are observed. Using qPCR, we show that rat hepatocytes express multiple purinergic P2Y and P2X receptors (R). ADP acting through P2Y1R elicits narrow Ca2+ oscillations, whereas UTP acting through P2Y2R elicits broad Ca2+ oscillations, with composite patterns observed for ATP. P2XRs do not play a role at physiological agonist levels. The discrete Ca2+ signatures reflect differential effects of protein kinase C (PKC), which selectively modifies the falling phase of the Ca2+ spikes. Negative feedback by PKC limits the duration of P2Y1R-induced Ca2+ spikes in a manner that requires extracellular Ca2+. By contrast, P2Y2R is resistant to PKC negative feedback. Thus, the PKC leg of the bifurcated IP3 signaling pathway shapes unique Ca2+ oscillation patterns that allows for distinct cellular responses to different agonists.

The absence of ‘shovel-ready’ anti-coronavirus drugs during vaccine development has exceedingly worsened the SARS-CoV-2 pandemic. Furthermore, new vaccine-resistant variants and coronavirus outbreaks may occur in the near future, and we must be ready to face this possibility. However, efficient antiviral drugs are still lacking to this day, due to our poor understanding of the mode of incorporation and mechanism of action of nucleotides analogs that target the coronavirus polymerase to impair its essential activity. Here, we characterize the impact of remdesivir (RDV, the only FDA-approved anti-coronavirus drug) and other nucleotide analogs (NAs) on RNA synthesis by the coronavirus polymerase using a high-throughput, single-molecule, magnetic-tweezers platform. We reveal that the location of the modification in the ribose or in the base dictates the catalytic pathway(s) used for its incorporation. We show that RDV incorporation does not terminate viral RNA synthesis, but leads the polymerase into backtrack as far as 30 nt, which may appear as termination in traditional ensemble assays. SARS-CoV-2 is able to evade the endogenously synthesized product of the viperin antiviral protein, ddhCTP, though the polymerase incorporates this NA well. This experimental paradigm is essential to the discovery and development of therapeutics targeting viral polymerases.

Parkinson’s Disease is the most common neurodegenerative movement disorder globally, with prevalence increasing. There is an urgent need for new therapeutics which are disease-modifying rather than symptomatic. Mitochondrial dysfunction is a well-documented mechanism in both sporadic and familial Parkinson’s Disease. Furthermore, ursodeoxycholic acid (UDCA) has been identified as a bile acid which leads to increased mitochondrial function in multiple in vitro and in vivo models of Parkinson’s Disease. Here, we describe the synthesis of novel C-nor-D-homo bile acid derivatives and the 12-hydroxy-methylated derivative of lagocholic acid (7) and their biological evaluation in fibroblasts from patients with either sporadic or LRRK2 mutant Parkinson’s Disease. These compounds boost mitochondrial function to a similar level or above that of UDCA in many assays; notable, however, is their ability to boost mitochondrial function to a higher level and at lower concentrations than UDCA specifically in the fibroblasts from LRRK2 patients. Our study indicates that novel bile acid chemistry could lead to the development of more efficacious bile acids which increase mitochondrial function and ultimately cellular health at lower concentrations proving attractive potential novel therapeutics for Parkinson’s Disease.

We report a first in-depth comparison of immune reconstitution in patients with HIV-related lymphoma following autologous hematopoietic cell transplant (AHCT) recipients (n=37, lymphoma, BEAM conditioning), HIV(-) AHCT recipients (n=30, myeloma, melphalan conditioning) at 56, 180, and 365 days post-AHCT, and 71 healthy control subjects. Principal component analysis showed that immune cell composition in HIV(+) and HIV(-) AHCT recipients clustered away from healthy controls and from each other at each time point, but approached healthy controls over time. Unsupervised feature importance score analysis identified activated T cells, cytotoxic memory and effector T cells [higher in HIV(+)], and naïve and memory T helper cells [lower HIV(+)] as a having a significant impact on differences between HIV(+) AHCT recipient and healthy control lymphocyte composition (p

This project investigated adding 8 weeks of biweekly plyometric and short sprints training into standard training in elite youth soccer players. An experimental group (EG, n = 18, age: 17.6 ± 0.6 years, body mass: 67.6 ± 5.8 kg, height: 1.75 ± 0.06 m, and body fat: 11.5 ± 1.6%) and control group (CG, n = 18, age: 17.5 ± 0.6 years, body mass: 68.8 ± 3.6 kg, height: 1.77 ± 0.04 m, and body fat: 11.7 ± 1.2%) participated. Pre-intervention and post-intervention measures were squat-jump (SJ), countermovement-jump (CMJ), standing long jump (SLJ), 5 and 20 m sprints, change-of-direction ability (4 × 5 m sprint test [S 4 × 5 m] and sprint 9-3-6-3-9 m with backward and forward running [SBF]), repeated change of direction (RCOD), and static balance (the stork balance test). For all parameters, significant (p < 0.001, ηp2 > 0.10) time and interaction (group × time) effects were observed. For three parameters (SBF, RCOD fastest time, and SLJ) no significant group effects were observed. The EG consistently showed a significantly higher performance level than the CG and a higher amount of effect sizes d (EG: drange: 1.27–2.61; CG: drange: 0.13–0.79) as an indicator for the development of performance between pre-intervention and post-intervention measures. Adding biweekly plyometric and short sprint training to standard training improves the athletic performance of young soccer players. Such plyometric and short sprint training conditioning can be highly recommended as part of the annual short training program for male elite under-19 (U19) soccer players.

This study explored the effects of home confinement on physical activity (PA) and satisfaction with life (SL) among university students during the COVID-19 pandemic. A total of 531 subjects participated [male: n=203; female: n=328; age: 33.1±5.2years; mass: 72.1±17.5kg; height: 1.67±0.12m; and body mass index (BMI): 25.7±5.06 kg/m2]. Online survey questions considered “before” and “during” confinement. Confinement reduced all PA intensities (ηp2=0.09–0.45, p

Increased basal low-grade inflammation is observed with advancing age, which is augmented by physical inactivity. However, data regarding the influence of lifelong exercise training and particularly high-intensity interval training (HIIT) on inflammatory mediators in older men are scarce. Therefore, we examined effects of 6weeks of aerobic preconditioning followed by 6weeks of HIIT on inflammatory mediators [interleukin (IL)-6, homocysteine, and high-sensitivity C-reactive protein (hsCRP)] in previously sedentary older men (SED) and masters athletes (LEX). Further, we investigated whether SED exhibited greater basal inflammatory biomarkers compared to LEX. Twenty-two men (aged 62±2years) participated in the SED group, while 17 age-matched LEX men (aged 60±5years) also participated as a positive comparison group. In SED, preconditioning (P=0.030, d=0.34) and HIIT (P=0.030, d=0.48) caused a reduction in IL-6 compared to enrollment. SED homocysteine did not change throughout (P>0.57; d0.42; d0.72), but homocysteine was not different (all P >0.131; d

Background: Sarcopenia is defined as a progressive and generalized loss of skeletal muscle quantity and function associated predominantly with aging. Physical activity appears the most promising intervention to attenuate sarcopenia, yet physical activity guidelines are rarely met. In recent years high intensity interval training (HIIT) has garnered interested in athletic populations, clinical populations, and general population alike. There is emerging evidence of the efficacy of HIIT in the young old (i.e. seventh decade of life), yet data concerning the oldest old (i.e., ninth decade of life onwards), and those diagnosed with sarcopenic are sparse.Objectives: In this scoping review of the literature, we aggregated information regarding HIIT as a potential intervention to attenuate phenotypic characteristics of sarcopenia.Eligibility Criteria: Original investigations concerning the impact of HIIT on muscle function, muscle quantity or quality, and physical performance in older individuals (mean age ≥60 years of age) were considered.Sources of Evidence: Five electronic databases (Medline, EMBASE, Web of Science, Scopus, and the Cochrane Central Register of Controlled Trials [CENTRAL]) were searched.Methods: A scoping review was conducted using the Arksey and O'Malley methodological framework (2005). Review selection and characterization were performed by two independent reviewers using pretested forms.Results: Authors reviewed 1,063 titles and abstracts for inclusion with 74 selected for full text review. Thirty-two studies were analyzed. Twenty-seven studies had a mean participant age in the 60s, two in the 70s, and three in the 80s. There were 20 studies which examined the effect of HIIT on muscle function, 22 which examined muscle quantity, and 12 which examined physical performance. HIIT was generally effective in Improving muscle function and physical performance compared to non-exercised controls, moderate intensity continuous training, or pre-HIIT (study design-dependent), with more ambiguity concerning muscle quantity.Conclusions: Most studies presented herein utilized outcome measures defined by the European Working Group on Sarcopenia in Older People (EWGSOP). However, there are too few studies investigating any form of HIIT in the oldest old (i.e., ≥80 years of age), or those already sarcopenic. Therefore, more intervention studies are needed in this population.

Abstract A new automated method to retrieve charge layer polarity from flashes, named Chargepol, is presented in this paper. Using data from the NASA Lightning Mapping Array (LMA) deployed during the Remote sensing of Electrification, Lightning, And Mesoscale/microscale Processes with Adaptive Ground Observations (RELAMPAGO) field campaign in Cordoba, Argentina, from November 2018 to April 2019, this method estimates the polarity of vertical charge distributions and their altitudes and thicknesses (or vertical depth) using the very‐high frequency (VHF) source emissions detected by LMAs. When this method is applied to LMA data for extended periods of time, it is capable of inferring a storm's bulk electrical charge structure throughout its life cycle. This method reliably predicted the polarity of charge within which lightning flashes propagated and was validated in comparison to methods that require manual assignment of polarities via visual inspection of VHF lightning sources. Examples of normal and anomalous charge structures retrieved using Chargepol for storms in Central Argentina during RELAMPAGO are presented for the first time. Application of Chargepol to five months of LMA data in Central Argentina and several locations in the United States allowed for the characterization of the charge structure in these regions and for a reliable comparison using the same methodology. About 13.3% of Cordoba thunderstorms were defined by an anomalous charge structure, slightly higher than in Oklahoma (12.5%) and West Texas (11.1%), higher than Alabama (7.3%), and considerably lower than in Colorado (82.6%). Some of the Cordoba anomalous thunderstorms presented enhanced low‐level positive charge, a feature rarely if ever observed in Colorado thunderstorms.

Humans first start to perceive the relationship between object size and mass in simple collision events at about 5.5–6.5 months of age. They perceive this link in simple collision events by attending to the size of the moving object and anticipating a greater displacement after collision with a large object and a lesser displacement with a small object. The results this aforementioned experiment is based on infants’ responses to a large and small object propelling a stationary object to the same distance (long distance). It is unknown how infants would perceive the same events if a large and small object propelled a stationary object to size appropriate (congruent) and size inappropriate (incongruent) distances. This paper aims to investigate this with adults (experiment 1) and 6-to-7-month-old infants (experiment 2). The first experiment served to validate our computer-generated collision events, by asking adults (N = 24) to rate the likeness of collision events happening in real-life, based on object size. In the second experiment, we tested this phenomenon in infants (N = 16) using the looking time paradigm. Results from the first experiment revealed that our computer-generated collision events are in line with adults’ assumptions of size-appropriate and size-inappropriate distances that the cube is propelled to by the small and large ball. Adults rated congruent test events as more likely than incongruent test events when asked how real-life-based they were. Results from the second experiment revealed infants distinguished between the sizes by preferring to look at the large ball longer than the small ball. However, the infants did not differ in their looking times for congruent and incongruent test events for small or/and large balls. For that reason, we conclude infants can distinguish between the sizes of the balls but are unable to perceive the size and mass associations in collision events.

The increasing usage of smart wearable devices has made an impact not only on the lifestyle of the users, but also on biological research and personalized healthcare services. These devices, which carry different types of sensors, have emerged as personalized digital diagnostic tools. Data from such devices have enabled the prediction and detection of various physiological as well as psychological conditions and diseases. In this review, we have focused on the diagnostic applications of wrist-worn wearables to detect multiple diseases such as cardiovascular diseases, neurological disorders, fatty liver diseases, and metabolic disorders, including diabetes, sleep quality, and psychological illnesses. The fruitful usage of wearables requires fast and insightful data analysis, which is feasible through machine learning. In this review, we have also discussed various machine-learning applications and outcomes for wearable data analyses. Finally, we have discussed the current challenges with wearable usage and data, and the future perspectives of wearable devices as diagnostic tools for research and personalized healthcare domains.

Q fever is caused by the bacterium Coxiella burnetii and is spread to humans from infected animals especially goats, sheep and cattle, predominantly when giving birth. There is an effective human vaccine (Q-VAX) against Q fever, and although Q fever is a worldwide problem, the vaccine is only used in Australia due to difficulties associated with its use and the risk of adverse reactions. The desire to protect humans, particularly farmers and abattoir workers, from Q fever prompted the development of a new safe and effective human vaccine without all the difficulties associated with the current vaccine. Candidate vaccines were prepared using purified O-specific polysaccharide (OSP) extracted from the lipopolysaccharide of virulent (phase 1) C. burnetii, strain Nine Mile, which was then conjugated to a tetanus toxoid (TT) carrier protein. Two vaccines were prepared using OSP from C. burnetii grown in embryonated eggs (vaccine A) and axenic media (vaccine B). Vaccines with or without alum adjuvant were used to vaccinate guinea pigs, which were later challenged by intranasal inoculation with virulent C. burnetii. Both vaccines protected guinea pigs from fever and loss of weight post challenge. Post-mortem samples of the spleen, liver and kidney of vaccinated guinea pigs contained substantially less C. burnetii DNA as measured by PCR than those of the unvaccinated control animals. This study demonstrated that a C. burnetii OSP-TT conjugate vaccine is capable of inducing protection against virulent C. burnetii in guinea pigs. Additionally, OSP derived from C. burnetii grown in axenic media compared to OSP from embryonated eggs is equivalent in terms of providing a protective immune response.

Adults judge darker objects to be heavier in weight than brighter objects, and objects which make lower pitch sounds as heavier in weight than objects making higher pitch sounds. It is unknown whether adults would make similar pairings if they saw these object properties in collision events. Two experiments examined adults’ judgements of computer-generated collision events based on object brightness and collision pitch sound. These experiments were designed as a precursor for an infant study, to validate the phenomenon. Results from the first experiment revealed that adults rated the bright ball likely event (where the bright ball displaced a stationary object a short distance after colliding with it) higher than the bright ball unlikely event. Conversely, adults rated the dark ball unlikely event (where the dark ball displaced a stationary object a short distance after colliding with it) higher than the dark ball likely event. Results from the second experiment demonstrated that adults judged the low pitch unlikely event (where the ball displaced a stationary object a short distance with a low pitch sound) higher than the low pitch likely event. Moreover, adults judged the high pitch likely event (where the ball displaced a stationary object a short distance with a high pitch sound) higher than the high pitch unlikely event. Results of these experiments suggest adults do not appropriately consider object brightness and pitch sound in collision events.

BackgroundTwo strategies of the US Ending the HIV Epidemic initiative are early diagnosis of infections via widespread testing and prevention of new infections using pre-exposure prophylaxis (PrEP). These strategies are particularly important for adolescents and young adults (AYAs) who are disproportionately affected by HIV, particularly if they identify as Black and/or lesbian, gay, bisexual, transgender, queer or questioning, and others (LGBTQ+). This study will develop and test an interactive life-simulation game in which players can enact real-life behaviors and receive their HIV risk profile to improve HIV testing and PrEP access among AYAs aged 13-24 years in Washington, DC. ObjectiveThis mixed methods study aims to determine the acceptability of an interactive, enhanced life-simulation game prototype among AYAs, conduct a pilot test of the gaming intervention among a small cohort of AYAs to ensure game usability and acceptability, and evaluate the efficacy of the game in a randomized controlled study with AYAs at risk for HIV in Washington, DC. MethodsThis research protocol will be conducted in 3 phases. A formative phase will involve surveys and focus groups (n=64) with AYAs living in the DC area. These focus groups will allow researchers to understand youth preferences for game enhancement. The second phase will consist of a pilot test (n=10) of the gaming intervention. This pilot test will allow researchers to modify the game based on formative results and test the planned recruitment and data collection strategy with intended end users. The third phase will consist of a randomized controlled study among 300 AYAs to examine the efficacy of the life-simulation game compared with app-based HIV educational materials on HIV and PrEP in changing HIV testing, knowledge, risk behaviors, and PrEP access. Participants will have unlimited access to either the life-simulation game or the educational app for 3 months from the time of enrollment. Study assessments will occur at enrollment and at 1, 3, and 6 months post enrollment via e-surveys. At 6 months, a subset of intervention participants (n=25) will participate in in-depth exit interviews regarding their experience being in the study. ResultsInstitutional review board approval was received on February 5, 2020. This project is currently recruiting participants for the formative phase. ConclusionsThis interactive life-simulation intervention aims to increase HIV testing and PrEP access among AYAs in the DC area. In this intervention, players can enact real-life behaviors and receive their HIV risk profile to promote HIV testing and PrEP seeking. Such an intervention has great potential to improve knowledge of HIV and PrEP among AYAs, increase motivation and self-efficacy related to HIV testing and PrEP use, and decrease individual and structural barriers that often preclude engagement in HIV prevention services. Trial RegistrationClinicalTrials.gov NCT04917575; https://clinicaltrials.gov/ct2/show/NCT04917575 International Registered Report Identifier (IRRID)PRR1-10.2196/29792

Policy responses to COVID-19 in the United States have been constrained by electoral calculi that run from the executive to the legislative branch of the central government and thence also to the federal bureaucracy and the states. The policy outcomes are as disappointing for US democracy as they are for the health of the US population.

Light-sheet microscopes are increasingly used for imaging cleared tissues, but have imposed constraints on sample geometries and protocols. Here the authors present a multi-immersion open-top light-sheet microscope to overcome these limitations and enable high-throughput imaging of samples processed with various clearing protocols.

Abstract Background The health impacts of community design have been studied extensively over the past two decades. In particular, public transportation use is associated with more walking between transit stops and shops, work, home and other destinations. Change in transit access has been linked with physical activity and obesity but seldom to health outcomes and associated costs, especially within a causal framework. Health related fiscal impacts of transit investment should be a key consideration in major transit investment decisions. Methods The Rails & Health study is a natural experiment evaluating changes in clinical measures, health care utilization and health care costs among Kaiser Permanente Northwest (KPNW) members following the opening of a new light rail transit (LRT) line in Portland, Oregon. The study is prospectively following 3036 adults exposed to the new LRT line and a similar cohort of 4386 adults who do not live close to the new line. Individual-level outcomes and covariates are extracted from the electronic medical record at KPNW, including member demographics and comorbidities, blood pressure, body mass index, lipids, glycosylated hemoglobin, and health care utilization and costs. In addition, participants are surveyed about additional demographics, travel patterns, physical activity (PA), and perceived neighborhood walkability. In a subsample of the study population, we are collecting direct measures of travel-related behavior—physical activity (accelerometry), global positioning system (GPS) tracking, and travel diaries—to document mechanisms responsible for observed changes in health outcomes and cost. Comprehensive measures of the built environment at baseline and after rail construction are also collected. Statistical analyses will (1) examine the effects of opening a new LRT line on chronic disease indicators, health care utilization, and health care costs and (2) evaluate the degree to which observed effects of the LRT line on health measures and costs are mediated by changes in total and transportation-associated PA. Discussion The results of the Rails & Health study will provide urban planners, transportation engineers, health practitioners, developers, and decision makers with critical information needed to document how transit investments impact population health and related costs.

Bile acid receptors have been identified as important targets for the development of new therapeutics to treat various metabolic and inflammatory diseases. The synthesis of new bile acid analogues can help elucidate structure–activity relationships and define compounds that activate these receptors selectively. Towards this, access to large quantities of a chenodeoxycholic acid derivative bearing a C-12 methyl and a C-13 to C-14 double bond provided an interesting scaffold to investigate the chemical manipulation of the C/D ring junction in bile acids. The reactivity of this alkene substrate with various zinc carbenoid species showed that those generated using the Furukawa methodology achieved selective α-cyclopropanation, whereas those generated using the Shi methodology reacted in an unexpected manner giving rise to a rearranged skeleton whereby the C ring has undergone contraction to form a novel spiro–furan ring system. Further derivatization of the cyclopropanated steroid included O-7 oxidation and epimerization to afford new bile acid derivatives for biological evaluation.