Search

Tissue-specific gene knockout by CRISPR/Cas9 is a powerful approach for characterizing gene functions during development. However, this approach has not been successfully applied to most Drosophila tissues, including the Drosophila neuromuscular junction (NMJ). To expand tissue-specific CRISPR to this powerful model system, here we present a CRISPR-mediated tissue-restricted mutagenesis (CRISPR-TRiM) toolkit for knocking out genes in motoneurons, muscles, and glial cells. We validated the efficacy of CRISPR-TRiM by knocking out multiple genes in each tissue, demonstrated its orthogonal use with the Gal4/UAS binary expression system, and showed simultaneous knockout of multiple redundant genes. We used CRISPR-TRiM to discover an essential role for SNARE components in NMJ maintenance. Furthermore, we demonstrate that the canonical ESCRT pathway suppresses NMJ bouton growth by downregulating retrograde Gbb signaling. Lastly, we found that axon termini of motoneurons rely on ESCRT-mediated intra-axonal membrane trafficking to release extracellular vesicles at the NMJ. Thus, we have successfully developed an NMJ CRISPR mutagenesis approach which we used to reveal genes important for NMJ structural plasticity. Author summary: We have developed a tissue-specific Cas9 toolkit that enables gene knockout specifically in motor neurons, glial cells, and muscle cells, the three principle cell types of the Drosophila peripheral motor system. Complementary to existing RNAi methods, this versatile tissue-specific knockout system offers unique advantages for dissecting gene functions at the neuromuscular junction (NMJ). Using these tools, we discovered that SNARE-mediated secretory pathways are required to maintain the integrity of the NMJ and that ESCRT components play critical yet differential roles in the biogenesis of extracellular vesicles, bouton growth, and membrane turnover at the NMJ. This CRISPR toolkit can be applied to study many biological questions in the neuromuscular system. [ABSTRACT FROM AUTHOR]

Patient-derived induced pluripotent stem cells (iPSCs) provide a powerful tool for identifying cellular and molecular mechanisms of disease. Macular telangiectasia type 2 (MacTel) is a rare, late-onset degenerative retinal disease with an extremely heterogeneous genetic architecture, lending itself to the use of iPSCs. Whole-exome sequencing screens and pedigree analyses have identified rare causative mutations that account for less than 5% of cases. Metabolomic surveys of patient populations and GWAS have linked MacTel to decreased circulating levels of serine and elevated levels of neurotoxic 1-deoxysphingolipids (1-dSLs). However, retina-specific, disease- contributing factors have yet to be identified. Here, we used iPSC-differentiated retinal pigmented epithelial (iRPE) cells derived from donors with or without MacTel to screen for novel cell-intrinsic pathological mechanisms. We show that MacTel iRPE cells mimicked the low serine levels observed in serum from patients with MacTel. Through RNA-Seq and gene set enrichment pathway analysis, we determined that MacTel iRPE cells are enriched in cellular stress pathways and dysregulation of central carbon metabolism. Using respirometry and mitochondrial stress testing, we functionally validated that MacTel iRPE cells had a reduction in mitochondrial function that was independent of defects in serine biosynthesis and 1-dSL accumulation. Thus, we identified phenotypes that may constitute alternative disease mechanisms beyond the known serine/sphingolipid pathway., Introduction Complex diseases of aging provide unique challenges for the identification of specific disease-causing genetic variants and associated cellular dysfunctions using traditional genetic approaches. Disease modeling using iPSCs is a [...]

Purpose: Reflex cough is an essential airway protective mechanism that often occurs in the context of divided attention. The effect of divided attention on measures of reflexive cough airflow and sensitivity remains unknown. We present findings from a study testing the effects of divided attention (via a dual-task paradigm) on measures of reflex cough in healthy young adults. Method: Volunteers (N = 20, age = 20-40 years) underwent 4 blocks of capsaicin-induced cough challenges. Within each block, capsaicin ranging from 0 to 200 [micro]M was presented in a randomized order. Two blocks consisted of cough testing only (single task). During the other 2 blocks, participants counted tones while simultaneously undergoing cough testing (dual task). Measures of cough motor response, self-reported urge-to-cough, cough frequency, and cough airflow were collected. Results: Participants coughed more in the single-task condition compared to the dual-task condition (p = 0.001). Participants' urge-to-cough ratings were lower in the dual-task condition (x¯ = 2, "slight") compared to the single-task condition (x¯ = 3, "moderate"; p = 0.007). Participants' cough reflex sensitivity thresholds were significantly increased in the dual-task condition (p = 0.002). Cough peak expiratory flow rates did not change between the 2 conditions (p = 0.34). Conclusions: Somatosensation of tussive stimuli changes during dual tasking. Abnormal cortical resource allocation may be a mechanism involved in silent aspiration.

While religion and spirituality (R/S) have been broadly studied for their positive mental health impacts, instances of abuse within religious or spiritual contexts remain under-researched. This scoping review aims to elucidate how individuals experiencing such abuse navigate their trauma, find meaning, and foster posttraumatic growth (PTG). The research was conducted using a scoping review methodology as a guide, and 10 articles were selected based on predefined inclusion and exclusion criteria. Synthesizing these articles revealed the following three central themes: recognizing abuse, relaying one's story, and redefining spirituality. Survivors often face disbelief and stigma, hindering their ability to process their experiences. However, narrative sharing enables many to reclaim agency and healing through validation and the integration of the narrative into one's life story. Additionally, survivors often transform spirituality, shifting from rigid frameworks to more nuanced and flexible understandings of the Divine and self. These findings underscore the importance of trauma-informed, spiritually sensitive clinical approaches that validate survivors' experiences, facilitate narrative sharing, and support spiritual redefining. Future research must address knowledge gaps, including the development of improved assessment tools, exploration of effective treatment strategies, and the unifying of terms to better support survivors' healing journeys and promote meaning-making and PTG in the aftermath of R/S abuse. [ABSTRACT FROM AUTHOR]

Background:In December 2021, the Region 1 Disaster Health Response System, the state of Vermont, and the National Emergency Tele-Critical Care Network partnered to provide statewide access to disaster teleconsultations during COVID-19 surge conditions. In this case report, we describe how a disaster teleconsultation system was implemented in Vermont to provide access to temporary tele-critical care consultations during the Omicron COVID-19 surge. Methods:We measured the time from request of service to implementation and calculated descriptive statistics. Results:Seven of Vermont's 14 hospitals requested the service. Despite a technology solution capable of providing services within hours, mean time to service implementation was 27 days (interquartile range 20–41 days). Conclusions:Integration of disaster teleconsultation systems into state and local emergency management plans are needed to bring administrative start-up times in line with technical readiness. [ABSTRACT FROM AUTHOR]

Identification of the key biotic and abiotic drivers within food webs is important for understanding species abundance changes in ecosystems, particularly across ecotones where there may be strong variation in interaction strengths. Using structural equation models (SEMs) and four decades of integrated data from the San Francisco Estuary, we investigated the relative effects of top‐down, bottom‐up, and environmental drivers on multiple trophic levels of the pelagic food web along an estuarine salinity gradient and at both annual and monthly temporal resolutions. We found that interactions varied across the estuarine gradient and that the detectability of different interactions depended on timescale. For example, for zooplankton and estuarine fishes, bottom‐up effects appeared to be stronger in the freshwater upstream regions, while top‐down effects were stronger in the brackish downstream regions. Some relationships (e.g., bottom‐up effects of phytoplankton on zooplankton) were seen primarily at annual timescales, whereas others (e.g., temperature effects) were only observed at monthly timescales. We also found that the net effect of environmental drivers was similar to or greater than bottom‐up and top‐down effects for all food web components. These findings can help identify which trophic levels or environmental factors could be targeted by management actions to have the greatest impact on estuarine forage fishes and the spatial and temporal scale at which responses might be observed. More broadly, this study highlights how environmental gradients can structure community interactions and how long‐term data sets can be leveraged to generate insights across multiple scales. [ABSTRACT FROM AUTHOR]

Purpose: This systematic review represents an update to previous reviews of the literature addressing behavioral management of respiratory/phonatory dysfunction in individuals with dysarthria due to neurodegenerative disease. Method: Multiple electronic database searches and hand searches of prominent speech-language pathology journals were conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses standards. Results: The search yielded 1,525 articles, from which 88 met inclusion criteria and were reviewed by two blinded co-investigators. A large range of therapeutic approaches have been added to the evidence base since the last review, including expiratory muscle strength training, singing, and computer- and device-driven programs, as well as a variety of treatment modalities, including teletherapy. Evidence for treatment in several different population groups--including cerebellar ataxia, myotonic dystrophy, autosomal recessive spastic ataxia of Charlevoix--Saguenay, Huntington's disease, multiple system atrophy, and Lewy body dementia--were added to the current review. Synthesis of evidence quality provided strong evidence in support of only one behavioral intervention: Lee Silverman Voice Treatment Program (LSVT LOUD) in people with Parkinson's disease. No other treatment approach or population included in this review demonstrated more than limited evidence, reflecting that these approaches/populations require urgent further examination. Conclusion: Suggestions about where future research efforts could be significantly strengthened and how clinicians can apply research findings to their practice are provided. [ABSTRACT FROM AUTHOR]

Textbooks are expensive, updated infrequently, and rarely used effectively by students. We discuss here a way for students to create the textbook for the course, helping them feel ownership over the course material. This Wiki-based, student-created textbook is online free for use, widely accessible by all, and editable during the course of and as topics evolve. This type of textbook format is particularly well suited to upper-level electives on topics that are rapidly emerging. We have nucleated a student-created textbook here, fully online and open access, for two upper elective courses in chemical engineering. Wikis offer an easy-to-learn platform that does not require previous training in coding, and we have found it to be an excellent way to increase student learning while encouraging student buy-in and ownership of their course textbook.

Purpose: Reflex cough is an essential airway protective mechanism that often occurs in the context of divided attention. The effect of divided attention on measures of reflexive cough airflow and sensitivity remains unknown. We present findings from a study testing the effects of divided attention (via a dual-task paradigm) on measures of reflex cough in healthy young adults. Method: Volunteers (N = 20, age = 20-40 years) underwent 4 blocks of capsaicin-induced cough challenges. Within each block, capsaicin ranging from 0 to 200 [micro]M was presented in a randomized order. Two blocks consisted of cough testing only (single task). During the other 2 blocks, participants counted tones while simultaneously undergoing cough testing (dual task). Measures of cough motor response, self-reported urge-to-cough, cough frequency, and cough airflow were collected. Results: Participants coughed more in the single-task condition compared to the dual-task condition (p [less than or equal to] .001). Participants' urge-to-cough ratings were lower in the dual-task condition ([bar.x] = 2, 'slight') compared to the single-task condition ([bar.x] = 3, 'moderate'; p = .007). Participants' cough reflex sensitivity thresholds were significantly increased in the dual-task condition (p = .002). Cough peak expiratory flow rates did not change between the 2 conditions (p = .34). Conclusions: Somatosensation of tussive stimuli changes during dual tasking. Abnormal cortical resource allocation may be a mechanism involved in silent aspiration., Reflex cough is an essential airway protective mechanism that occurs in response to penetration and aspiration of foods and liquids. Airway protection and coughing must often take place in the [...]