Remote Teaching of Publication-Quality, Single-Case Graphs in Microsoft Excel

Microsoft Excel is ubiquitous, cost-effective, and can be used to create publication-quality single-case design graphs. We systematically replicated the GraphPad Prism video tutorial by Mitteer et al. (2018) to teach 24 master\u27s students to create multiple-baseline graphs using Excel 2016. Students\u27 mean accuracy on the multiple-baseline graph was 25% in pretraining, 86% with the video tutorial, and 96% with the review checklist. Next, students used the same video tutorial to create multielement and reversal graphs. Students\u27 mean accuracy on the multielement graph was 93% with video tutorial and 94% with review checklist, and accuracy on the reversal graph was 82% with video tutorial and 94% with review checklist. Students reported moderate to high satisfaction with both training components. The results support scientist-practitioners using the video tutorial and review checklists to create three common graphs using Excel 2016, Excel 2019, and Excel Office 365

SARS-CoV-2 infection of the pancreas promotes thrombofibrosis and is associated with new-onset diabetes

Evidence suggests an association between severe acute respiratory syndrome-cornavirus-2 (SARS-CoV-2) infection and the occurrence of new-onset diabetes. We examined pancreatic expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), the cell entry factors for SARS-CoV-2, using publicly available single-cell RNA sequencing data sets, and pancreatic tissue from control male and female nonhuman primates (NHPs) and humans. We also examined SARS-CoV-2 immunolocalization in pancreatic cells of SARS-CoV-2-infected NHPs and patients who had died from coronavirus disease 2019 (COVID-19). We report expression of ACE2 in pancreatic islet, ductal, and endothelial cells in NHPs and humans. In pancreata from SARS-CoV-2-infected NHPs and COVID-19 patients, SARS-CoV-2 infected ductal, endothelial, and islet cells. These pancreata also exhibited generalized fibrosis associated with multiple vascular thrombi. Two out of 8 NHPs developed new-onset diabetes following SARS-CoV-2 infection. Two out of 5 COVID-19 patients exhibited new-onset diabetes at admission. These results suggest that SARS-CoV-2 infection of the pancreas may promote acute and especially chronic pancreatic dysfunction that could potentially lead to new-onset diabetes

Treatment of Porphyromonas gulae infection and downstream pathology in the aged dog by lysine-gingipain inhibitor COR388.

COR388, a small-molecule lysine-gingipain inhibitor, is currently being investigated in a Phase 2/3 clinical trial for Alzheimer's disease (AD) with exploratory endpoints in periodontal disease. Gingipains are produced by two species of bacteria, Porphyromonas gingivalis and Porphyromonas gulae, typically associated with periodontal disease and systemic infections in humans and dogs, respectively. P. gulae infection in dogs is associated with periodontal disease, which provides a physiologically relevant model to investigate the pharmacology of COR388. In the current study, aged dogs with a natural oral infection of P. gulae and periodontal disease were treated with COR388 by oral administration for up to 90 days to assess lysine-gingipain target engagement and reduction of bacterial load and downstream pathology. In a 28-day dose-response study, COR388 inhibited the lysine-gingipain target and reduced P. gulae load in saliva, buccal cells, and gingival crevicular fluid. The lowest effective dose was continued for 90 days and was efficacious in continuous reduction of bacterial load and downstream periodontal disease pathology. In a separate histology study, dog brain tissue showed evidence of P. gulae DNA and neuronal lysine-gingipain, demonstrating that P. gulae infection is systemic and spreads beyond its oral reservoir, similar to recent observations of P. gingivalis in humans. Together, the pharmacokinetics and pharmacodynamics of COR388 lysine-gingipain inhibition, along with reduction of bacterial load and periodontal disease in naturally occurring P. gulae infection in the dog, support the use of COR388 in targeting lysine-gingipain and eliminating P. gingivalis infection in humans

Syntaphilin controls a mitochondrial rheostat for proliferation-motility decisions in cancer.

Tumors adapt to an unfavorable microenvironment by controlling the balance between cell proliferation and cell motility, but the regulators of this process are largely unknown. Here, we show that an alternatively spliced isoform of syntaphilin (SNPH), a cytoskeletal regulator of mitochondrial movements in neurons, is directed to mitochondria of tumor cells. Mitochondrial SNPH buffers oxidative stress and maintains complex II-dependent bioenergetics, sustaining local tumor growth while restricting mitochondrial redistribution to the cortical cytoskeleton and tumor cell motility. Conversely, introduction of stress stimuli to the microenvironment, including hypoxia, acutely lowered SNPH levels, resulting in bioenergetics defects and increased superoxide production. In turn, this suppressed tumor cell proliferation but increased tumor cell invasion via greater mitochondrial trafficking to the cortical cytoskeleton. Loss of SNPH or expression of an SNPH mutant lacking the mitochondrial localization sequence resulted in increased metastatic dissemination in xenograft or syngeneic tumor models in vivo. Accordingly, tumor cells that acquired the ability to metastasize in vivo constitutively downregulated SNPH and exhibited higher oxidative stress, reduced cell proliferation, and increased cell motility. Therefore, SNPH is a stress-regulated mitochondrial switch of the cell proliferation-motility balance in cancer, and its pathway may represent a therapeutic target

Measuring pH of the Coxiella burnetii Parasitophorous Vacuole

Coxiella burnetii is the causative agent of human Q fever, a zoonotic disease that can cause a debilitating, flu‐like illness in acute cases, or a life‐threatening endocarditis in chronic patients. An obligate intracellular bacterial pathogen, Coxiella survives and multiplies in a large lysosome‐like vacuole known as the Coxiella parasitophorous vacuole (CPV). A unique characteristic of the CPV is the acidic environment (pH ∼5.0), which is required to activate Coxiella metabolism and the Coxiella type 4 secretion system (T4SS), a major virulence factor required for intracellular survival. Further, inhibiting or depleting vacuolar ATPase, a host cell protein that regulates lysosomal pH, inhibits intracellular Coxiella growth. Together, these data suggest that CPV pH is an important limiting factor for Coxiella growth and virulence. This unit describes a method to determine CPV pH using live cell microscopy of a pH–sensitive fluorophore conjugated to dextran. This technique is useful to measure changes in CPV pH during infection or in response to drug treatment

Enhanced Written Instructions for Creating Publication-Quality Single-Case Design Graphs in Microsoft Excel

Graphs are visual descriptors of functional relations between behavior and the environment, and behavioral researchers and clinicians make informed decisions on current and future procedures by evaluating such functional relations. Thus, graphically depicting and visually inspecting single-subject data is foundational in the science of behavior. Microsoft Excel is the most prevalent program used by behavior analysts to create single-case design graphs; however, the Excel literature mostly includes brief tutorials and descriptions of its utility, neither of which evaluate methods of training Excel’s comprehensive capabilities. Self-directed training with enhanced written instructions (EWI) is a viable option to train graphing skills as it alleviates the amount of resources required by in-person trainings. However, published evaluations of EWI as a method to train graphing are limited in quantity, rely on permanent product measures, and exclude assessments of maintenance and generalization. We used a multiple baseline across participants design to evaluate the effects of EWI to train seven undergraduate students to create publication-quality single-subject design graphs in Excel. We measured graphing accuracy and latency to graph completion using real-time, live-Excel, and permanent product measures. We also assessed response maintenance and generalization. EWI resulted in immediate, robust effects, and we observed generalization and maintenance across all participants. We discuss these results and their implications regarding staff training, Excel’s utility, and data measurement

Drug-tolerant persister cancer cells are vulnerable to GPX4 inhibition.

Acquired drug resistance prevents cancer therapies from achieving stable and complete responses. Emerging evidence implicates a key role for non-mutational drug resistance mechanisms underlying the survival of residual cancer 'persister' cells. The persister cell pool constitutes a reservoir from which drug-resistant tumours may emerge. Targeting persister cells therefore presents a therapeutic opportunity to impede tumour relapse. We previously found that cancer cells in a high mesenchymal therapy-resistant cell state are dependent on the lipid hydroperoxidase GPX4 for survival. Here we show that a similar therapy-resistant cell state underlies the behaviour of persister cells derived from a wide range of cancers and drug treatments. Consequently, we demonstrate that persister cells acquire a dependency on GPX4. Loss of GPX4 function results in selective persister cell ferroptotic death in vitro and prevents tumour relapse in mice. These findings suggest that targeting of GPX4 may represent a therapeutic strategy to prevent acquired drug resistance

Teaching Graphing Using Enhanced Written Instructions: Does Chunk Size Matter

Graphing is an important feature of the field of behavior analysis, not only as a job responsibility of behavioral professionals, but as a visual analysis tool as well. While graphing can be taught using various methods, perhaps self-training methods could prove both effective and efficient due to the self-guided nature of the methods. One effective self-training method for graphing is enhanced written instructions (EWI). While the literature has demonstrated EWI’s effectiveness when training graphing, specific presentations of EWI have not been evaluated. To address this gap in the literature, we compared the graphing accuracy and duration to graph completion of chunked presentations of EWI, and evaluated preference for the two different chunked presentations, using a nonconcurrent multiple baseline design across five students with various degrees of graphing history. Both chunked presentations were found to be effective, with most participants clearly preferring one presentation over the other

Thermal weed control in Photinia x Fraseri “Red Robin” container nurseries

A near-zero tolerance policy on weeds by markets for nursery crops calls for weed-free container-grown plants, and forces growers to frequently remove weeds. Thermal weed control could represent a novel method to control weeds in shrubs from container nurseries, thus avoiding the use of herbicides and mulches. The aims of this study were to develop custom-built machinery for thermal weed control in container nurseries and to test the weed control efficiency of flame weeding and steaming in Photinia x fraseri "Red Robin" containers. A liquefied petroleum gas (LPG) fed flamer and a steamer with a dedicated diffuser were built. Four treatments were applied for a total period of 24 months: steaming once every four months, steaming once every two months, flame weeding once every two months or once a month. Temperature values measured at different depths in the substrate after thermal applications were recorded and analyzed. Photinia x fraseri features (height, diameter, and dry biomass) and aesthetic parameters as affected by thermal treatments were also evaluated. The trend in temperature values of the substrate over time followed a two-phase exponential decay. All the thermal treatments lead to a continuous near-100% weed control level, which is the level required by growers for aesthetic reasons. No damages caused by heat on Photinia x fraseri were observed. Container nursery producers could thus adopt thermal methods as a substitute for chemical solutions for weed control management