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There are few pharmacologic options for the treatment of cognitive deficits associated with traumatic brain injury in pediatric patients. Acetylcholinesterase inhibitors such as donepezil have been evaluated in adult patients after traumatic brain injury, but relatively less is known about the effect in pediatric populations. The goal of this review is to identify knowledge gaps in the efficacy and safety of acetylcholinesterase inhibitors as a potential adjuvant treatment for neurocognitive decline in pediatric patients with traumatic brain injury. Investigators queried PubMed to identify literature published from database inception through June 2023 describing the use of donepezil in young adult traumatic brain injury and pediatric patients with predefined conditions. Based on preselected search criteria, 340 unique papers were selected for title and abstract screening. Thirty-two records were reviewed in full after eliminating preclinical studies and papers outside the scope of the project. In adult traumatic brain injury, we review results from 14 papers detailing 227 subjects where evidence suggests donepezil is well tolerated and shows both objective and patient-reported efficacy for reducing cognitive impairment. In children, 3 papers report on 5 children recovering from traumatic brain injury, showing limited efficacy. An additional 15 pediatric studies conducted in populations at risk for cognitive dysfunction provide a broader look at safety and efficacy in 210 patients in the pediatric age group. Given its promise for efficacy in adults with traumatic brain injury and tolerability in pediatric patients, we believe further study of donepezil for children and adolescents with traumatic brain injury is warranted.

Abstract Anorexia nervosa (AN) and atypical AN (AtypAN) are complex neurobiological illnesses that typically onset in adolescence with an often treatment-refractory and chronic illness trajectory. Aberrant eating behaviors in this population have been linked to abnormalities in food reward and cognitive control, but prior studies have not examined respective contributions of clinical characteristics and metabolic state. Research is needed to identify specific disruptions and inform novel intervention targets to improve outcomes. Fifty-nine females with AN (n = 34) or AtypAN (n = 25), ages 10–22 years, all ≤90% expected body weight, and 34 age-matched healthy controls (HC) completed a well-established neuroimaging food cue paradigm fasting and after a standardized meal, and we used ANCOVA models to investigate main and interaction effects of Group and Appetitive State on blood oxygenation level-dependent (BOLD) activation for the contrast of exposure to high-calorie food images minus objects. We found main effects of Group with greater BOLD activation in the dorsal anterior cingulate cortex (dACC), dorsolateral prefrontal cortex (DLPFC), hippocampus, caudate, and putamen for AN/AtypAN versus HC groups, and in the three-group model including AN, AtypAN, and HC (sub-)groups, where differences were primarily driven by greater activation in the AtypAN subgroup versus HC group. We found a main effect of Appetitive State with increased premeal BOLD activation in the hypothalamus, amygdala, nucleus accumbens, and caudate for models that included AN/AtypAN and HC groups, and in BOLD activation in the nucleus accumbens for the model that included AN, AtypAN, and HC (sub-)groups. There were no interaction effects of Group with Appetitive State for any of the models. Our findings demonstrate robust feeding-state independent group effects reflecting greater neural activation of specific regions typically associated with reward and cognitive control processing across AN and AtypAN relative to healthy individuals in this food cue paradigm. Differential activation of specific brain regions in response to the passive viewing of high-calorie food images may underlie restrictive eating behavior in this clinical population.

Abstract Wildlife translocations are increasingly used to combat declining biodiversity worldwide. Successful translocation often hinges on coexistence between humans and wildlife, yet not all translocation efforts explicitly include human dimensions (e.g., economic incentives, education programs, and conflict reduction assistance). To evaluate the prevalence and associated outcomes of including human dimensions as objectives when planning translocations, we analyze 305 case studies from the IUCN’s Global Re-Introduction Perspectives Series. We find that fewer than half of all projects included human dimension objectives (42%), but that projects including human dimension objectives were associated with improved wildlife population outcomes (i.e., higher probability of survival, reproduction, or population growth). Translocation efforts were more likely to include human dimension objectives if they involved mammals, species with a history of local human conflict, and local stakeholders. Our findings underscore the importance of incorporating objectives related to human dimensions in translocation planning efforts to improve conservation success.

Liu et al. used the NASA GRACE/FO missions to show that since 2019, groundwater depletion in California’s Central Valley has accelerated by 31% compared to recent droughts, and has increased by a nearly a factor of 5 compared to the 60-year average.

The first wave of the COVID-19 pandemic led to significant socioeconomic changes in Canada due to business and school closures, and related job losses. This increased food insecurity among vulnerable populations, as well as many who had not been previously food insecure, placing unprecedented demand on charitable food organizations. This study documented the pandemic’s impact on charitable food organizations in Manitoba, Canada during the first wave in spring 2020. Using a multi-method design, data on pandemic-related program challenges and newly implemented policies/procedures were collected from: food bank organization websites and Facebook pages; online news media outlets; and semi-structured interviews with food organization leadership. Inductive thematic analysis was used to identify emerging patterns and themes. Second level coding was used to integrate data from different sources. Six challenge themes emerged: increased need for services; acquisition and distribution of food supply; staff and volunteer resource management; emotional vulnerability of staff, volunteers, and clients; difficulties with internal and external communications; and lack of structural supports. Five policy/procedure themes emerged: program and service delivery changes; finance and administrative changes; safety protocols; advocacy for resources and community engagement; and changes to paid and volunteer staffing. The first wave of COVID-19 had a significant impact on the Manitoba charitable food sector. Food banks re-configured programs to meet client needs amid shifting public health directives, with diminished resources, rising demand, and insufficient government support. Despite the resiliency of community food organizations during the pandemic, the status quo with respect to addressing food insecurity is inefficient and inadequate.

Abstract Objectives The COVID-19 pandemic has impacted all aspects of the food system, including the retail grocery sector. We sought to (objective 1) document and (objective 2) analyze the policies implemented in the grocery sector during the first wave of the pandemic in Manitoba, Canada. Methods Our qualitative policy analysis draws from organizational communications (websites and social media) (n = 79), news media articles (n = 95), and key informant interviews with individuals (n = 8) working within the grocery sector in urban and rural, Manitoba. Media and communications were extracted between March 9-May 8, 2020 and interviews were conducted in July–August, 2020. Results Newly implemented policies due to the pandemic fell under four inter-related themes: Employee health and wellbeing, Safety measures, Operational measures, and Community support. Employee health and wellbeing included sub-themes of financial and social support, health recommendations and protocols, and new employee guidelines. Safety measures encompassed numerous policies pertaining to sanitation, personal protection, transmission prevention, physical distancing, and limiting access. Overall, new policies were discussed as effective in making grocery shopping as safe as possible given the situation. Compliance and enforcement, employee teamwork, and support for employees were key themes related to perceptions of policy success in a challenging and inequitable context. Nevertheless, government support and communication was needed as well to ensure safety within the grocery sector. Conclusions The grocery sector reacted to the pandemic with the swift implementation of policies to address food supply issues, prevent transmission of the virus, support their employees as essential workers, and better serve high-risk populations.

The membrane biofilm reactor (MBfR), which is based on the counter diffusion of the electron donors and acceptors into the biofilm, represents a novel technology for wastewater treatment. When process air or oxygen is supplied, the MBfR is known as the membrane aerated biofilm reactor (MABR), which has high oxygen transfer rate and efficiency, promoting microbial growth and activity within the biofilm. Over the past few decades, laboratory-scale studies have helped researchers and practitioners understand the relevance of influencing factors and biological transformations in MABRs. In recent years, pilot- to full-scale installations are increasing along with process modeling. The resulting accumulated knowledge has greatly improved understanding of the counter-diffusional biological process, with new challenges and opportunities arising. Therefore, it is crucial to provide new insights by conducting this review. This paper reviews wastewater treatment advancements using MABR technology, including design and operational considerations, microbial community ecology, and process modeling. Treatment performance of pilot- to full-scale MABRs for process intensification in existing facilities is assessed. This paper also reviews other emerging applications of MABRs, including sulfur recovery, industrial wastewater, and xenobiotics bioremediation, space-based wastewater treatment, and autotrophic nitrogen removal. In conclusion, commercial applications demonstrate that MABR technology is beneficial for pollutants (COD, N, P, xenobiotics) removal, resource recovery (e.g., sulfur), and N2O mitigation. Further research is needed to increase packing density while retaining efficient external mass transfer, understand the microbial interactions occurring, address existing assumptions to improve process modeling and control, and optimize the operational conditions with site-specific considerations. HIGHLIGHTS Performance of commercial MABR applications is assessed.; Design and operational considerations are evaluated for both pure and hybrid MABRs.; Microbial community ecology in the counter-diffusional membrane-aerated biofilm is reviewed.; Review covers current state-of-the-art process modeling for MABR studies.; Existing challenges are identified to assist with framing future opportunities for MABR technology.;

Abstract Background Anorexia nervosa (AN) is associated with structural brain abnormalities. Studies have reported less cerebral tissue and more cerebrospinal fluid (CSF) in individuals with AN relative to healthy controls, although findings are variable and inconsistent due to variations in sample size, age, and disease state (e.g., active AN, weight-recovered AN). Further, it remains unclear if structural brain abnormalities observed in AN are a consequence of specific brain pathologies or malnutrition, as very few longitudinal neuroimaging studies in AN have been completed. Methods To overcome this issue, this comprehensive meta-analysis will combine region-of-interest (ROI) and voxel-based morphometry (VBM) approaches to understand how regional and global structural brain abnormalities differ among individuals with AN and healthy controls (HCs). Additionally, we aim to understand how clinical characteristics and physiological changes during the course of illness, including acute illness vs. weight recovery, may moderate these structural abnormalities. We will create an online database of studies that have investigated structural brain abnormalities in AN. Data will be reviewed independently by two members of our team using MEDLINE databases, Web of Science, PsycINFO, EMBASE, and CINAHL. We will conduct ROI and VBM meta-analysis using seed-based d mapping in AN and HCs. We will include all studies that include structural neuroimaging of individuals with AN (both acute and weight-recovered) and HCs between January 1997 and 2020. Discussion This systematic review will assess the effects of AN compared to HC on brain structure. Futhermore, it will explore the role of acute AN and weight-recovered AN on brain structure. Findings will help researchers and clinicians to better understand the course of illness in AN and the nature of recovery, in terms of weight, malnutrition, and the state of the brain. Systematic review registration PROSPERO CRD42020180921

Abstract In buzz-pollinated plants, bees apply thoracic vibrations to the flower, causing pollen release from anthers, often through apical pores. Bees grasp one or more anthers with their mandibles, and vibrations are transmitted to this focal anther(s), adjacent anthers, and the whole flower. Pollen release depends on anther vibration, and thus it should be affected by vibration transmission through flowers with distinct morphologies, as found among buzz-pollinated taxa. We compare vibration transmission between focal and non-focal anthers in four species with contrasting stamen architectures: Cyclamen persicum, Exacum affine, Solanum dulcamara and S. houstonii. We used a mechanical transducer to apply bee-like vibrations to focal anthers, measuring the vibration frequency and displacement amplitude at focal and non-focal anther tips simultaneously using high-speed video analysis (6000 frames per second). In flowers in which anthers are tightly arranged (C. persicum and S. dulcamara), vibrations in focal and non-focal anthers are indistinguishable in both frequency and displacement amplitude. In contrast, flowers with loosely arranged anthers (E. affine) including those with differentiated stamens (heterantherous S. houstonii), show the same frequency but higher displacement amplitude in non-focal anthers compared to focal anthers. We suggest that stamen architecture modulates vibration transmission, potentially affecting pollen release and bee behaviour.

Abstract Background Robot-assisted platforms in spine surgery have rapidly developed into an attractive technology for both the surgeon and patient. Although current literature is promising, more clinical data is needed. The purpose of this paper is to determine the effect of robot-related complications on clinical outcomes Methods This multicenter study included adult (≥18 years old) patients who underwent robot-assisted lumbar fusion surgery from 2012-2019. The minimum follow-up was 1 year after surgery. Both bivariate and multivariate analyses were performed to determine if robot-related factors were associated with reoperation within 1 year after primary surgery. Results A total of 320 patients were included in this study. The mean (standard deviation) Charlson Comorbidity Index was 1.2 (1.2) and 52.5% of patients were female. Intraoperative robot complications occurred in 3.4% of patients and included intraoperative exchange of screw (0.9%), robot abandonment (2.5%), and return to the operating room for screw exchange (1.3%). The 1-year reoperation rate was 4.4%. Robot factors, including robot time per screw, open vs. percutaneous, and robot system, were not statistically different between those who required revision surgery and those who did not (P>0.05). Patients with robot complications were more likely to have prolonged length of hospital stay and blood transfusion, but were not at higher risk for 1-year reoperations. The most common reasons for reoperation were wound complications (2.2%) and persistent symptoms due to inadequate decompression (1.5%). In the multivariate analysis, robot related factors and complications were not independent risk factors for 1-year reoperations. Conclusion This is the largest multicenter study to focus on robot-assisted lumbar fusion outcomes. Our findings demonstrate that 1-year reoperation rates are low and do not appear to be influenced by robot-related factors and complications; however, robot-related complications may increase the risk for greater blood loss requiring a blood transfusion and longer length of stay.

BACKGROUND COVID-19 remains a global health emergency with limited treatment options, lagging vaccine rates, and inadequate healthcare resources in the face of an ongoing calamity. The disease is characterized by immune dysregulation and cytokine storm. Cyclosporine A (CSA) is a calcineurin inhibitor that modulates cytokine production and may have direct antiviral properties against coronaviruses.METHODS To test whether a short course of CSA was safe in patients with COVID-19, we treated 10 hospitalized, oxygen-requiring, noncritically ill patients with CSA (starting at a dose of 9 mg/kg/d). We evaluated patients for clinical response and adverse events, measured serum cytokines and chemokines associated with COVID-19 hyperinflammation, and conducted gene-expression analyses.RESULTS Five participants experienced adverse events, none of which were serious; transaminitis was most common. No participant required intensive care unit–level care, and all patients were discharged alive. CSA treatment was associated with significant reductions in serum cytokines and chemokines important in COVID-19 hyperinflammation, including CXCL10. Following CSA administration, we also observed a significant reduction in type I IFN gene expression signatures and other transcriptional profiles associated with exacerbated hyperinflammation in the peripheral blood cells of these patients.CONCLUSION Short courses of CSA appear safe and feasible in patients with COVID-19 who require oxygen and may be a useful adjunct in resource-limited health care settings.TRIAL REGISTRATION This trial was registered on ClinicalTrials.gov (Investigational New Drug Application no. 149997; ClinicalTrials.gov NCT04412785).FUNDING This study was internally funded by the Center for Cellular Immunotherapies.

Mathematical modelling was used to investigate the possibility to use membrane aerated biofilm reactors (MABRs) in a largely anoxic suspended growth bioreactor to produce the nitrate-nitrogen required for heterotrophic denitrification and the growth of denitrifying phosphorus accumulating organisms (DPAOs). The results indicate that such a process can be used to achieve a variety of process objectives. The capture of influent biodegradable organic matter while also achieving significant total inorganic nitrogen (TIN) removal can be achieved with or without use of primary treatment by operation at a relatively short suspended growth solids residence time (SRT). Low effluent TIN concentrations can also be achieved, irrespective of the influent wastewater chemical oxygen demand (COD)/total nitrogen (TN) ratio, with somewhat larger suspended growth SRT. Biological phosphorus and nitrogen removal can also be effectively achieved. Further experimental work is needed to confirm these modelling results. HIGHLIGHTS Hybrid MABRs can achieve lower effluent TIN concentrations, and more carbon capture, at lower SRTs than conventional systems.; Influent carbon composition affects denitrification, but modelling shows better performance in hybrid MABR systems.; Primary treatment benefits carbon capture, but can be eliminated depending on treatment goals.; Combined biological nitrogen and phosphorus removal is possible and improved in hybrid MABRs.;

Objective This was a retrospective, cohort study investigating the efficacy and safety of continuous low-dose postoperative tranexamic acid (PTXA) on drain output and transfusion requirements following adult spinal deformity surgery. Methods One hundred forty-seven patients undergoing posterior instrumented thoracolumbar fusion of ≥ 3 vertebral levels at a single institution who received low-dose PTXA infusion (0.5–1 mg/kg/hr) for 24 hours were compared to 292 control patients who did not receive PTXA. The cohorts were propensity matched based on age, sex, American Society of Anesthesiologist physical status classification, body mass index, number of surgical levels, revision surgery, operative duration, and total intraoperative TXA dose (n = 106 in each group). Primary outcome was 72-hour postoperative drain output. Secondary outcomes were number of allogeneic blood transfusions. Results There was no significant difference in postoperative drain output in the PTXA group compared to control (660 ±420 mL vs. 710 ±490 mL, p = 0.46). The PTXA group received significantly more crystalloid (6,100 ±3,100 mL vs. 4,600 ±2,400 mL, p < 0.001) and red blood cell transfusions postoperatively (median [interquartile range]: 1 [0–2] units vs. 0 [0–1] units; incidence rate ratio [95% confidence interval], 1.6 [1.2–2.2]; p = 0.001). Rates of adverse events were comparable between groups. Conclusion Continuous low-dose PTXA infusion was not associated with reduced drain output after spinal deformity surgery. No difference in thromboembolic incidence was observed. A prospective dose escalation study is warranted to investigate the efficacy of higher dose PTXA.

Abstract Global declines in wildlife migrations have prompted new initiatives to conserve remaining migratory behaviors. However, many migrations have already been lost. Important attempts have been made to recover extirpated migrations, and our understanding of restoration remains narrowly confined to these particular species and landscapes. Here, we examine diverse restoration efforts through the unifying lens of behavioral ecology to draw broader inferences regarding the feasibility and effectiveness of restoring lost migrations. First, we synthesize recent research advances that illuminate key roles of exploration, learning, and adaptation in migratory behavior. Then, we review case studies to identify common themes of restoration success across four major vertebrate groups: fish, birds, mammals, and herpetofauna. We describe three broad strategies to effectively restore lost migrations: reestablishing migratory populations, recovering migratory habitats, and reviving migratory behavior itself. To guide conservation and research efforts, we link these strategies with specific management techniques, and we explore the biological mechanisms underpinning the success of each. Our work reveals a previously underappreciated potential for restoring lost migrations in terrestrial and freshwater vertebrates, and it provides guidance on whether and how conservation practitioners, researchers, and policymakers can work to restore the valuable migrations we have lost.

From the combined perspective of biologists, microscope instrumentation developers, imaging core facility scientists, and high performance computing experts, we discuss the challenges faced when selecting imaging and analysis tools in the field of light-sheet microscopy. Our goal is to provide a contextual framework of basic computing concepts that cell and developmental biologists can refer to when mapping the peculiarities of different light-sheet data to specific existing computing environments and image analysis pipelines. We provide our perspective on efficient processes for tool selection and review current hardware and software commonly used in light-sheet image analysis, as well as discuss what ideal tools for the future may look like.

Mutualisms involve cooperation, but also frequently involve conflict. Plant-pollinator mutualisms are no exception. To facilitate animal pollination, flowering plants often offer pollen (their male gametes) as a food reward. Since plants benefit by maximizing pollen export to conspecific flowers, we might expect plants to cheat on pollen rewards. In intersexual floral mimicry, rewarding pollen-bearing male flowers (models) are mimicked by rewardless female flowers (mimics) on the same plant. Pollinators should therefore learn to avoid the unrewarding mimics. Plants might impede such learning by producing phenotypically variable flowers that cause bees to generalize among models and mimics during learning. In this laboratory study, we used partially artificial flowers (artificial petals, live reproductive parts) modeled after Begonia odorata to test whether variation in the size of rewarding male flowers (models) and unrewarding female flowers (mimics) affected how quickly bees learned both to recognize models and to reject mimics. Live unrewarding female flowers have 33% longer petals and have 31% greater surface area than live rewarding male flowers, which bees should easily discriminate. Yet while bees rapidly learned to reduce foraging effort on mimics, learning was not significantly affected by the degree to which flower size varied. Additionally, we found scant evidence that this was a result of bees altering response speed to maintain decision accuracy. Our study failed to provide evidence that flower size variation in intersexual floral mimicry systems exploits pollinator cognition, though we cannot rule out that other floral traits that are variable may be important. Furthermore, we propose that contrary to expectation, phenotypic variability in a Batesian mimicry system may not necessarily have significant effects on whether receivers effectively learn to discriminate models and mimics.

Phytocannabinoids, such as Δ9-tetrahydrocannabinol (THC), bind and activate cannabinoid (CB) receptors, thereby “piggy-backing” on the same pathway's endogenous endocannabinoids (ECs). The recent discovery that liver fatty acid binding protein-1 (FABP1) is the major cytosolic “chaperone” protein with high affinity for both Δ9-THC and ECs suggests that Δ9-THC may alter hepatic EC levels. Therefore, the impact of Δ9-THC or EC treatment on the levels of endogenous ECs, such as N-arachidonoylethanolamide (AEA) and 2-arachidonoylglycerol (2-AG), was examined in cultured primary mouse hepatocytes from WT and Fabp1 gene-ablated (LKO) mice. Δ9-THC alone or 2-AG alone significantly increased AEA and especially 2-AG levels in WT hepatocytes. LKO alone markedly increased AEA and 2-AG levels. However, LKO blocked/diminished the ability of Δ9-THC to further increase both AEA and 2-AG. In contrast, LKO potentiated the ability of exogenous 2-AG to increase the hepatocyte level of AEA and 2-AG. These and other data suggest that Δ9-THC increases hepatocyte EC levels, at least in part, by upregulating endogenous AEA and 2-AG levels. This may arise from Δ9-THC competing with AEA and 2-AG binding to FABP1, thereby decreasing targeting of bound AEA and 2-AG to the degradative enzymes, fatty acid amide hydrolase and monoacylglyceride lipase, to decrease hydrolysis within hepatocytes.

Upregulation of the hepatic endocannabinoid (EC) receptor [cannabinoid receptor-1 (CB1)] and arachidonoylethanolamide (AEA) is associated with nonalcoholic fatty liver disease (NAFLD). Male mice fed high-fat diet (HFD) ad libitum also exhibit NAFLD, increased hepatic AEA, and obesity. But, preference for HFD complicates interpretation and almost nothing is known about these effects in females. These issues were addressed by pair-feeding HFD. Similarly to ad libitum-fed HFD, pair-fed HFD also increased WT male and female mouse fat tissue mass (FTM), but preferentially at the expense of lean tissue mass. In contrast, pair-fed HFD did not elicit NAFLD in WT mice regardless of sex. Concomitantly, pair-fed HFD oppositely impacted hepatic AEA, 2-arachidonoyl glycerol, and/or CB1 in WT males versus females. In pair-fed HFD mice, liver FA binding protein-1 (Fabp1) gene ablation (LKO): i) exacerbated FTM in both sexes; ii) did not elicit liver neutral lipid accumulation in males and only slightly in females; iii) increased liver AEA in males, but decreased it in females; and iv) decreased CB1 only in males. Thus, pair-fed HFD selectively impacted hepatic ECs more in females, but did not elicit NAFLD in either sex. These effects were modified by LKO consistent with FABP1's ability to impact EC and FA metabolism.

Studies in vitro have suggested that both sterol carrier protein-2/sterol carrier protein-x (Scp-2/Scp-x) and liver fatty acid binding protein [Fabp1 (L-FABP)] gene products facilitate hepatic uptake and metabolism of lipotoxic dietary phytol. However, interpretation of physiological function in mice singly gene ablated in the Scp-2/Scp-x has been complicated by concomitant upregulation of FABP1. The work presented herein provides several novel insights: i) An 8-anilino-1-naphthalenesulfonic acid displacement assay showed that neither SCP-2 nor L-FABP bound phytol, but both had high affinity for its metabolite, phytanic acid; ii) GC-MS studies with phytol-fed WT and Fabp1/Scp-2/SCP-x gene ablated [triple KO (TKO)] mice showed that TKO exacerbated hepatic accumulation of phytol metabolites in vivo in females and less so in males. Concomitantly, dietary phytol increased hepatic levels of total long-chain fatty acids (LCFAs) in both male and female WT and TKO mice. Moreover, in both WT and TKO female mice, dietary phytol increased hepatic ratios of saturated/unsaturated and polyunsaturated/monounsaturated LCFAs, while decreasing the peroxidizability index. However, in male mice, dietary phytol selectively increased the saturated/unsaturated ratio only in TKO mice, while decreasing the peroxidizability index in both WT and TKO mice. These findings suggested that: 1) SCP-2 and FABP1 both facilitated phytol metabolism after its conversion to phytanic acid; and 2) SCP-2/SCP-x had a greater impact on hepatic phytol metabolism than FABP1.

Abstract Dispersal is central to the ecology and evolution of spatially structured communities. While flower microbial communities are spatially structured among floral organs, how dispersal vectors distribute microbes among floral organs is unknown. Pollinators are recognized as key microbial vectors, but effects of their different foraging behaviors on transfer dynamics among flowers or different floral organs are not known. We asked how foraging behaviors of a model pollinator (Bombus impatiens) affect acquisition and dispersal of microbes among flower organs. We used monkeyflowers (Mimulus guttatus) to examine dispersal within a natural context and artificial flowers to test how common bee foraging behaviors (nectaring, buzzing, or scrabbling) shaped dispersal of a green fluorescent protein‐labeled bacteria, Pseudomonas fluorescens. Bees acquired 1% of a flower's microbes and dispersed 31% of acquired microbes to the next flower. All bees acquired microbes, and 85% and 76% of bees dispersed microbes to live and artificial flowers, respectively. Microbes acquired from the corolla were mainly deposited on the corolla, followed by the stamens, and least on the nectary/pistil. Bee foraging behavior affected acquisition, with scrabbling for pollen resulting in 23% more microbes acquired than nectaring, and with buzzing for pollen resulting in a 79% slower rate of microbial acquisition relative to scrabbling. Bee foraging behavior also affected deposition but depended on the floral organ: Scrabbling and buzzing for pollen led to greater deposition than nectaring for corolla and stamen but not nectary. Our results have implications for transmission of beneficial and pathogenic microbes among plants and pollinators, and thus the ecology and evolution of floral microbial communities.

Objective: Osteoporosis is increasing in incidence as the ageing population continues to grow. Decreased bone mineral density poses a challenge for the spine surgeon. In patients requiring lumbar interbody fusion, differences in diagnostics and surgical approaches may be warranted. In this systematic review, the authors examine studies performing lumbar interbody fusion in patients with osteopenia or osteoporosis and suggest avenues for future study. Methods: A systematic literature review of the PubMed and MEDLINE databases was performed for studies published between 1986 and 2020. Studies evaluating diagnostics, surgical approaches, and other technical considerations were included. Results: A total of 13 articles were ultimately selected for qualitative analysis. This includes studies demonstrating the utility of Hounsfield units in diagnosis, a survey of surgical approaches, as well as exploring the use of vertebral augmentation and cortical bone screw trajectory. Conclusions: This systematic review provides a summary of preliminary findings with respect to the use of Hounsfield units as a diagnostic tool, the benefit or lack thereof with respect to minimally invasive approaches, and the question of whether or not cement augmentation or cortical bone trajectory confers benefit in osteoporotic patients undergoing lumbar interbody fusion. While the findings of these studies are promising, the current state of the literature is limited in scope and, for this reason, definitive conclusions cannot be drawn from these data. The authors highlight gaps in the literature and the need for further exploration and study of lumbar interbody fusion in the osteoporotic spine.

Wildlife translocations are increasingly used to combat declining biodiversity worldwide. Successful translocation often hinges on coexistence between humans and wildlife, yet not all translocation efforts explicitly include human dimensions (e.g., economic incentives, education programs, and conflict reduction assistance). To evaluate the prevalence and associated outcomes of including human dimensions as objectives when planning translocations, we analyze 305 case studies from the IUCN's Global Re-Introduction Perspectives Series. We find that fewer than half of all projects included human dimension objectives (42%), but that projects including human dimension objectives were associated with improved wildlife population outcomes (i.e., higher probability of survival, reproduction, or population growth). Translocation efforts were more likely to include human dimension objectives if they involved mammals, species with a history of local human conflict, and local stakeholders. Our findings underscore the importance of incorporating objectives related to human dimensions in translocation planning efforts to improve conservation success.

Although lipid-rich microdomains of hepatocyte plasma membranes serve as the major scaffolding regions for cholesterol transport proteins important in cholesterol disposition, little is known regarding intracellular factors regulating cholesterol distribution therein. On the basis of its ability to bind cholesterol and alter hepatic cholesterol accumulation, the cytosolic liver type FA binding protein (L-FABP) was hypothesized to be a candidate protein regulating these microdomains. Compared with wild-type hepatocyte plasma membranes, L-FABP gene ablation significantly increased the proportion of cholesterol-rich microdomains. Lack of L-FABP selectively increased cholesterol, phospholipid (especially phosphatidylcholine), and branched-chain FA accumulation in the cholesterol-rich microdomains. These cholesterol-rich microdomains are important, owing to enrichment therein of significant amounts of key transport proteins involved in uptake of cholesterol [SR-B1, ABCA-1, P-glycoprotein (P-gp), sterol carrier binding protein (SCP-2)], FA transport protein (FATP), and glucose transporters 1 and 2 (GLUT1, GLUT2) insulin receptor. L-FABP gene ablation enhanced the concentration of SCP-2, SR-B1, FATP4, and GLUT1 in the cholesterol-poor microdomains, with functional implications in HDL-mediated uptake and efflux of cholesterol. Thus L-FABP gene ablation significantly impacted the proportion of cholesterol-rich versus -poor microdomains in the hepatocyte plasma membrane and altered the distribution of lipids and proteins involved in cholesterol uptake therein.

While plasma membrane cholesterol-rich microdomains play a role in cholesterol trafficking, little is known about the appearance and dynamics of cholesterol through these domains in living cells. The fluorescent cholesterol analog 6-dansyl-cholestanol (DChol), its biochemical fractionation, and confocal imaging of L-cell fibroblasts contributed the following new insights: i) fluorescence properties of DChol were sensitive to microenvironment polarity and mobility; (ii) DChol taken up by L-cell fibroblasts was distributed similarly as cholesterol and preferentially into cholesterol-rich vs. -poor microdomains resolved by affinity chromatography of purified plasma membranes; iii) DChol reported similar polarity (dielectric constant near 18) but higher mobility near phospholipid polar head group region for cholesterol in purified cholesterol-rich versus -poor microdomains; and iv) real-time confocal imaging, quantitative colocalization analysis, and fluorescence resonance energy transfer with cholesterol-rich and -poor microdomain markers confirmed that DChol preferentially localized in plasma membrane cholesterol-rich microdomains of living cells. Thus, DChol sensed a unique, relatively more mobile microenvironment for cholesterol in plasma membrane cholesterol-rich microdomains, consistent with the known, more rapid exchange dynamics of cholesterol from cholesterol-rich than -poor microdomains.

Although studies with liver type fatty acid binding protein (L-FABP) gene ablated mice demonstrate a physiological role for L-FABP in hepatic fatty acid metabolism, little is known about the mechanisms whereby L-FABP elicits these effects. Studies indicate that L-FABP may function to shuttle lipids to the nucleus, thereby increasing the availability of ligands of nuclear receptors, such as peroxisome proliferator-activated receptor-α (PPARα). The data herein suggest that such mechanisms involve direct interaction of L-FABP with PPARα. L-FABP was shown to directly interact with PPARα in vitro through co-immunoprecipitation (co-IP) of pure proteins, altered circular dichroic (CD) spectra, and altered fluorescence spectra. In vitro fluorescence resonance energy transfer (FRET) between Cy3-labeled PPARα and Cy5-labeled L-FABP proteins showed that these proteins bound with high affinity (Kd approximately 156 nM) and in close proximity (intermolecular distance of 52Å). This interaction was further substantiated by co-IP of both proteins from liver homogenates of wild-type mice. Moreover, double immunogold electron microscopy and FRET confocal microscopy of cultured primary hepatocytes showed that L-FABP was in close proximity to PPARα (intermolecular distance 40–49Å) in vivo. Taken together, these studies were consistent with L-FABP regulating PPARα transcriptional activity in hepatocytes through direct interaction with PPARα. Our in vitro and imaging experiments demonstrate high affinity, structural molecular interaction of L-FABP with PPARα and suggest a functional role for L-FABP interaction with PPARα in long chain fatty acid (LCFA) metabolism.

Although in vitro studies suggest a role for sterol carrier protein-2 (SCP-2) in cholesterol trafficking and metabolism, the physiological significance of these observations remains unclear. This issue was addressed by examining the response of mice overexpressing physiologically relevant levels of SCP-2 to a cholesterol-rich diet. While neither SCP-2 overexpression nor cholesterol-rich diet altered food consumption, increased weight gain, hepatic lipid, and bile acid accumulation were observed in wild-type mice fed the cholesterol-rich diet. SCP-2 overexpression further exacerbated hepatic lipid accumulation in cholesterol-fed females (cholesterol/cholesteryl esters) and males (cholesterol/cholesteryl esters and triacyglycerol). Primarily in female mice, hepatic cholesterol accumulation induced by SCP-2 overexpression was associated with increased levels of LDL-receptor, HDL-receptor scavenger receptor-B1 (SR-B1) (as well as PDZK1 and/or membrane-associated protein 17 kDa), SCP-2, liver fatty acid binding protein (L-FABP), and 3α-hydroxysteroid dehydrogenase, without alteration of other proteins involved in cholesterol uptake (caveolin), esterification (ACAT2), efflux (ATP binding cassette A-1 receptor, ABCG5/8, and apolipoprotein A1), or oxidation/transport of bile salts (cholesterol 7α-hydroxylase, sterol 27α-hydroxylase, Na+/taurocholate cotransporter, Oatp1a1, and Oatp1a4). The effects of SCP-2 overexpression and cholesterol-rich diet was downregulation of proteins involved in cholesterol transport (L-FABP and SR-B1), cholesterol synthesis (related to sterol regulatory element binding protein 2 and HMG-CoA reductase), and bile acid oxidation/transport (via Oapt1a1, Oatp1a4, and SCP-x). Levels of serum and hepatic bile acids were decreased in cholesterol-fed SCP-2 overexpression mice, especially in females, while the total bile acid pool was minimally affected. Taken together, these findings support an important role for SCP-2 in hepatic cholesterol homeostasis.

Although reverse cholesterol transport from peripheral cell types is mediated through plasma membrane microdomains termed lipid rafts, almost nothing is known regarding the existence, protein/lipid composition, or structure of these putative domains in liver hepatocytes, cells responsible for the net removal of cholesterol from the body. Lipid rafts purified from hepatocyte plasma membranes by a nondetergent affinity chromatography method were: i) present at 33 ± 3% of total plasma membrane protein; ii) enriched in key proteins of the reverse cholesterol pathway [scavenger receptor class B type I (SR-B1), ABCA1, P-glycoprotein (P-gp), sterol carrier protein-2 (SCP-2)]; iii) devoid of caveolin-1; iv) enriched in cholesterol, sphingomyelin, GM1, and phospholipids low in polyunsaturated fatty acid and double bond index; and v) exhibited an intermediate liquid-ordered lipid phase with significant transbilayer fluidity gradient. Ablation of the gene encoding SCP-2 significantly altered lipid rafts to: i) increase the proportion of lipid rafts present, thereby increasing raft total content of ABCA1, P-gp, and SR-B1; ii) increase total phospholipids while decreasing GM1 in lipid rafts; iii) decrease the fluidity of lipid rafts, consistent with the increased intermediate liquid-ordered phase; and iv) abolish the lipid raft transbilayer fluidity gradient. Thus, despite the absence of caveolin-1 in liver hepatocytes, lipid rafts represented nearly one-third of the mouse hepatocyte plasma membrane proteins and displayed unique protein, lipid, and biophysical properties that were differentially regulated by SCP-2 expression.

Despite the importance of branched chain lipid oxidation in detoxification, almost nothing is known regarding factors regulating peroxisomal uptake, targeting, and metabolism. One peroxisomal protein, sterol carrier protein-x (SCP-x), is thought to catalyze a key thiolytic step in branched chain lipid oxidation. When mice with substantially lower hepatic levels of SCP-x were tested for susceptibility to dietary stress with phytol (a phytanic acid precursor and peroxisome proliferator), livers of phytol-fed female but not male mice i) accumulated phytol metabolites (phytanic acid, pristanic acid, and Δ-2,3-pristanic acid); ii) exhibited decreased fat tissue mass and increased liver mass/body mass; iii) displayed signs of histopathological lesions in the liver; and iv) demonstrated significant alterations in hepatic lipid distributions. Moreover, both male and female mice exhibited phytol-induced peroxisomal proliferation, as demonstrated by liver morphology and upregulation of the peroxisomal protein catalase. In addition, levels of liver fatty acid binding protein, along with SCP-2 and SCP-x, increased, suggesting upregulation mediated by phytanic acid, a known ligand agonist of the peroxisomal proliferator-activated receptor α.In summary, the present work establishes a role for SCP-x in branched chain lipid catabolism and demonstrates a sexual dimorphic response to phytol, a precursor of phytanic acid, in lipid parameters and hepatotoxicity.

The purpose of this collective instrumental case study was to understand the motivational factors that support families' decisions to exhibit livestock. The expectancy value theory served as the theoretical lens, and a review of literature led to four issues to be explored. Four typical family cases were identified, and interviews were conducted. Five In-Vivo themes were identified: (a) "a family tradition," (b) "bonds us together," (c) "on-the-job training for life," (d) "joys and discomforts of agricultural life," and (e) "the show industry." It was concluded that families value tradition, family togetherness, the agricultural community, work ethic, and the development of life skills critical for the success of their children. Winning, as traditionally defined in the show ring, was not the expectation. Rather, families expected to grow together, enjoy their time, and be competitive. The perceived family utility outweighed the noted costs. It was recommended that all stakeholders in the livestock exhibition community identify ways to enhance family involvement and work to reward ethical behaviors.

Although a link between excess lipid storage and aberrant glucose metabolism has been recognized for many years, little is known what role lipid storage droplets and associated proteins such as Plin2 play in managing cellular glucose levels. To address this issue, the influence of Plin2 on glucose uptake was examined using 2-NBD-Glucose and [(3)H]-2-deoxyglucose to show that insulin-mediated glucose uptake was decreased 1.7- and 1.8-fold, respectively in L cell fibroblasts overexpressing Plin2. Conversely, suppression of Plin2 levels by RNAi-mediated knockdown increased 2-NBD-Glucose uptake several fold in transfected L cells and differentiated 3T3-L1 cells. The effect of Plin2 expression on proteins involved in glucose uptake and transport was also examined. Expression of the SNARE protein SNAP23 was increased 1.6-fold while levels of syntaxin-5 were decreased 1.7-fold in Plin2 overexpression cells with no significant changes observed in lipid droplet associated proteins Plin1 or FSP27 or with the insulin receptor, GLUT1, or VAMP4. FRET experiments revealed a close proximity of Plin2 to SNAP23 on lipid droplets to within an intramolecular distance of 51 Å. The extent of targeting of SNAP23 to lipid droplets was determined by co-localization and co-immunoprecipitation experiments to show increased partitioning of SNAP23 to lipid droplets when Plin2 was overexpressed. Taken together, these results suggest that Plin2 inhibits glucose uptake by interacting with, and regulating cellular targeting of SNAP23 to lipid droplets. In summary, the current study for the first time provides direct evidence for the role of Plin2 in mediating cellular glucose uptake.

While TOFA (acetyl CoA carboxylase inhibitor) and C75 (fatty acid synthase inhibitor) prevent lipid accumulation by inhibiting fatty acid synthesis, the mechanism of action is not simply accounted for by inhibition of the enzymes alone. Liver fatty acid binding protein (L-FABP), a mediator of long chain fatty acid signaling to peroxisome proliferator-activated receptor-α (PPARα) in the nucleus, was found to bind TOFA and its activated CoA thioester, TOFyl-CoA, with high affinity while binding C75 and C75-CoA with lower affinity. Binding of TOFA and C75-CoA significantly altered L-FABP secondary structure. High (20 mM) but not physiological (6 mM) glucose conferred on both TOFA and C75 the ability to induce PPARα transcription of the fatty acid β-oxidative enzymes CPT1A, CPT2, and ACOX1 in cultured primary hepatocytes from wild-type (WT) mice. However, L-FABP gene ablation abolished the effects of TOFA and C75 in the context of high glucose. These effects were not associated with an increased cellular level of unesterified fatty acids but rather by increased intracellular glucose. These findings suggested that L-FABP may function as an intracellular fatty acid synthesis inhibitor binding protein facilitating TOFA and C75-mediated induction of PPARα in the context of high glucose at levels similar to those in uncontrolled diabetes.

The purpose of this photovoice study was to investigate what and how pre-service teachers conceive and make meaning of exploration observations of early field experiences (EFEs). EFEs are vital components of the teacher preparation program and include all activities that occur in pre-service teacher education prior to student teaching, including exploratory observations (Retallick & Miller, 2010). Process coding of 123 photographs and reflective captions led to five themes: (a) optimizing student aperture, (b) affirming the decision to teach, (c) identifying learning strategy outcomes, (d) balancing the three components of agricultural education, and (e) creating a felt need to learn. Praxis included more purposeful inclusion of cooperating teachers as experts, distinguishing between an experience and observation, and seeking those observations outside of agricultural education classrooms.

Pollen collection is necessary for bee survival and important for flowering plant reproduction, yet if and how pollen extraction motor routines are modified with experience is largely unknown. Here, we used an automated reward and monitoring system to evaluate modification in a common pollen-extraction routine, floral sonication. Through a series of laboratory experiments with the bumblebee, Bombus impatiens, we examined whether variation in sonication frequency and acceleration is due to instrumental learning based on rewards, a fixed behavioral response to rewards, and/or a mechanical constraint. We first investigated whether bees could learn to adjust their sonication frequency in response to pollen rewards given only for specified frequency ranges and found no evidence of instrumental learning. However, we found that absence versus receipt of a pollen reward did lead to a predictable behavioral response, which depended on bee size. Finally, we found some evidence of mechanical constraints, in that flower mass affected sonication acceleration (but not frequency) through an interaction with bee size. In general, larger bees showed more flexibility in sonication frequency and acceleration, potentially reflecting a size-based constraint on the range over which smaller bees can modify frequency and acceleration. Overall, our results show that although bees did not display instrumental learning of sonication frequency, their sonication motor routine is nevertheless flexible.