Sensory systems rely on neuromodulators, such as serotonin, to provide flexibility for information processing as stimuli vary, such as light intensity throughout the day. Serotonergic neurons broadly ...innervate the optic ganglia of Drosophila melanogaster, a widely used model for studying vision. It remains unclear whether serotonin modulates the physiology of interneurons in the optic ganglia. To address this question, we first mapped the expression patterns of serotonin receptors in the visual system, focusing on a subset of cells with processes in the first optic ganglion, the lamina. Serotonin receptor expression was found in several types of columnar cells in the lamina including 5-HT2B in lamina monopolar cell L2, required for spatiotemporal luminance contrast, and both 5-HT1A and 5-HT1B in T1 cells, whose function is unknown. Subcellular mapping with GFP-tagged 5-HT2B and 5-HT1A constructs indicated that these receptors localize to layer M2 of the medulla, proximal to serotonergic boutons, suggesting that the medulla neuropil is the primary site of serotonergic regulation for these neurons. Exogenous serotonin increased basal intracellular calcium in L2 terminals in layer M2 and modestly decreased the duration of visually induced calcium transients in L2 neurons following repeated dark flashes, but otherwise did not alter the calcium transients. Flies without functional 5-HT2B failed to show an increase in basal calcium in response to serotonin. 5-HT2B mutants also failed to show a change in amplitude in their response to repeated light flashes but other calcium transient parameters were relatively unaffected. While we did not detect serotonin receptor expression in L1 neurons, they, like L2, underwent serotonin-induced changes in basal calcium, presumably via interactions with other cells. These data demonstrate that serotonin modulates the physiology of interneurons involved in early visual processing in Drosophila.
The Serotonin Transporter (SERT) regulates extracellular serotonin levels and is the target of most current drugs used to treat depression. The mechanisms by which inhibition of SERT activity ...influences behavior are poorly understood. To address this question in the model organism Drosophila melanogaster, we developed new loss of function mutations in Drosophila SERT (dSERT). Previous studies in both flies and mammals have implicated serotonin as an important neuromodulator of sleep, and our newly generated dSERT mutants show an increase in total sleep and altered sleep architecture that is mimicked by feeding the SSRI citalopram. Differences in daytime versus nighttime sleep architecture as well as genetic rescue experiments unexpectedly suggest that distinct serotonergic circuits may modulate daytime versus nighttime sleep. dSERT mutants also show defects in copulation and food intake, akin to the clinical side effects of SSRIs and consistent with the pleomorphic influence of serotonin on the behavior of D. melanogaster. Starvation did not overcome the sleep drive in the mutants and in male dSERT mutants, the drive to mate also failed to overcome sleep drive. dSERT may be used to further explore the mechanisms by which serotonin regulates sleep and its interplay with other complex behaviors.
Abstract
Dashboard-mounted touchscreen tablets are now common in vehicles. Screen/phone use in cars likely shifts drivers’ attention away from the road and contributes to risk of accidents. ...Nevertheless, vision is subject to multisensory influences from other senses. Haptics may help maintain or even increase visual attention to the road, while still allowing for reliable dashboard control. Here, we provide a proof-of-concept for the effectiveness of digital haptic technologies (hereafter digital haptics), which use ultrasonic vibrations on a tablet screen to render haptic perceptions. Healthy human participants (N = 25) completed a divided-attention paradigm. The primary task was a centrally-presented visual conjunction search task, and the secondary task entailed control of laterally-presented sliders on the tablet. Sliders were presented visually, haptically, or visuo-haptically and were vertical, horizontal or circular. We reasoned that the primary task would be performed best when the secondary task was haptic-only. Reaction times (RTs) on the visual search task were fastest when the tablet task was haptic-only. This was not due to a speed-accuracy trade-off; there was no evidence for modulation of VST accuracy according to modality of the tablet task. These results provide the first quantitative support for introducing digital haptics into vehicle and similar contexts.
Objectives
To identify clear cell renal cell carcinoma‐related gene mutations potentially associated with aggressive disease, sarcomatoid differentiation or poor prognosis.
Methods
We carried out ...genomic analysis of 217 tumor foci from 25 patients with conventional clear cell renal cell carcinoma (14 patients), clear cell renal cell carcinoma with sarcomatoid differentiation (six patients) and non‐clear cell renal cell carcinoma (five patients). Each tumor nodule on the tissue block that corresponded to the same focus on the slide was separated from the normal parenchyma and other histologically distinct areas of tumor. The isolated tumor foci were used for subsequent analyses and sequencing. Deoxyribonucleic acid from the formalin‐fixed paraffin‐embedded tissues was extracted. Multiplex bar‐coded polymerase chain reaction amplification was carried out using next‐generation sequencing libraries.
Results
Overall, 67 protein alterations, including amino acid alterations, frame shifts and splice site mutations in seven genes were identified in the cohort of renal cell carcinoma tumors included in this study. Fewer patients with clear cell renal cell carcinoma with sarcomatoid differentiation had clear cell renal cell carcinoma‐related mutations in comparison with patients with conventional clear cell renal cell carcinoma. Additionally, the average number of unique clear cell renal cell carcinoma‐related protein alterations per patient was significantly lower in clear cell renal cell carcinoma with sarcomatoid differentiation than in conventional clear cell renal cell carcinoma. Mutations in PBRM1 were identified in a higher proportion of patients with high‐grade tumors (World Health Organization/International Society of Urological Pathology grade 4) and in the primary tumors of six of 10 (60%) patients with metastatic disease.
Conclusions
Although there are pitfalls due to intratumoral heterogeneity and sampling bias, mutations in PBRM1 may be associated with metastasis and aggressive disease in clear cell renal cell carcinoma.
Abstract Exome sequencing (ES) has emerged as an essential tool in the evaluation of neurodevelopmental disorders (NDD) of unknown etiology. Genome sequencing (GS) offers advantages over ES due to ...improved detection of structural, copy number, repeat number and non‐coding variants. However, GS is less commonly utilized due to higher cost and more intense analysis. Here, we present nine cases of pediatric NDD that were molecularly diagnosed with GS between 2017 and 2022, following non‐diagnostic ES. All individuals presented with global developmental delay or regression. Other features present in our cohort included epilepsy, white matter abnormalities, brain malformation and dysmorphic features. Two cases were diagnosed on GS due to newly described gene‐disease relationship or variant reclassification ( MAPK8IP3, CHD3 ). Additional features missed on ES that were later detected on GS were: intermediate‐size deletions in three cases who underwent ES that were not validated for CNV detection, pathogenic variants within the non‐protein coding genes SNORD118 and RNU7‐1 , pathogenic variant within the promoter region of GJB1 , and a coding pathogenic variant within BCAP31 which was not sufficiently covered on ES. GS following non‐diagnostic ES led to the identification of pathogenic variants in this cohort of nine cases, four of which would not have been identified by reanalysis alone.
Melanin is a free‐radical scavenger, antioxidant, and broadband absorber of ultraviolet (UV) radiation which protects the skin from environmental carcinogenesis. However, melanin synthesis and ...UV‐induced reactive melanin species are also implicated in melanocyte genotoxicity. Here, we attempted to reconcile these disparate functions of melanin using a UVB‐sensitive, NRAS‐mutant mouse model, TpN. We crossed TpN mice heterozygous for an inactivating mutation in Tyrosinase to produce albino and black littermates on a C57BL/6J background. These animals were then exposed to a single UVB dose on postnatal day three when keratinocytes in the skin have yet to be melanized. Approximately one‐third (35%) of black mice were protected from UVB‐accelerated tumor formation. However, melanoma growth rates, tumor mutational burdens, and gene expression profiles were similar in melanomas from black and albino mice. Skin from albino mice contained more cyclobutane pyrimidine dimer (CPD) positive cells than black mice 1‐h post‐irradiation. However, this trend gradually reversed over time with CPDs becoming more prominent in black than albino melanocytes at 48 h. These results show that in the absence of epidermal pigmentation, melanocytic melanin limits the tumorigenic effects of acute UV exposure but fails to protect melanocytes from UVB‐induced mutagenesis.
New, conformationally restricted Th super(IV) and U super(IV) complexes, ThCl sub(2)(L) and UI sub(2)(L), of the small-cavity, dipyrrolide, dianionic macrocycle trans-calix2benzene2pyrrolide (L) ...super(2-) are reported and are shown to have unusual Kappa super(5): Kappa super(5) binding in a bent metallocene-type structure. Single-electron reduction of UI sub(2)(L) affords UI(THF)(L) and results in a switch in ligand binding from Kappa super(5)-pyrrolide to eta super(6)-arene sandwich coordination, demonstrating the preference for arene binding by the electron-rich U super(III) ion. Facile loss of THF from UI(THF)(L) further increases the amount of U-arene back donation. UI(L) can incorporate a further U super(III) equivalent, UI sub(3), to form the very unusual dinuclear complex U sub(2)I sub(4)(L) in which the single macrocycle adopts both Kappa super(5): Kappa super(5) and eta super(6): Kappa super(1): eta super(6): Kappa super(1) binding modes in the same complex. Hybrid density functional theory calculations carried out to compare the electronic structures and bonding of U super(III)I(L) and U super(III) sub(2)I sub(4)(L) indicate increased contributions to the covalent bonding in U sub(2)I sub(4)(L) than in UI(L), and similar U-arene interactions in both. MO analysis and QTAIM calculations find minimal U-U interaction in U sub(2)I sub(4)(L). In contrast to the reducible U complex, treatment of ThCl sub(2)(L) with either a reductant or non-nucleophilic base results in metallation of the aryl rings of the macrocycle to form the (L super(-2H)) super(4-) tetraanion and two new and robust Th-C bonds in the -ate complexes K(THF) sub(2)Th super(IV)( mu -Cl)(L super(-2H)) sub(2) and KTh super(IV){N(SiMe sub(3)) sub(2)}(L super(-2H)).
Problem: This study was designed to develop a deeper understanding of the learning and social processes that take place during the simulation-based medical education for practicing providers as part ...of the Project ECHO® model, known as Mock ECHO training. The ECHO model is utilized to expand access to care of common and complex diseases by supporting the education of primary care providers with an interprofessional team of specialists via videoconferencing networks. Intervention: Mock ECHO trainings are conducted through a train the trainer model targeted at leaders replicating the ECHO model at their organizations. Trainers conduct simulated teleECHO clinics while participants gain skills to improve communication and self-efficacy. Context: Three focus groups, conducted between May 2015 and January 2016 with a total of 26 participants, were deductively analyzed to identify common themes related to simulation-based medical education and interdisciplinary education. Principal themes generated from the analysis included (a) the role of empathy in community development, (b) the value of training tools as guides for learning, (c) Mock ECHO design components to optimize learning, (d) the role of interdisciplinary education to build community and improve care delivery, (e) improving care integration through collaboration, and (f) development of soft skills to facilitate learning. Outcome: Mock ECHO trainings offer clinicians the freedom to learn in a noncritical environment while emphasizing real-time multidirectional feedback and encouraging knowledge and skill transfer. Lessons Learned: The success of the ECHO model depends on training interprofessional healthcare providers in behaviors needed to lead a teleECHO clinic and to collaborate in the educational process. While building a community of practice, Mock ECHO provides a safe opportunity for a diverse group of clinician experts to practice learned skills and receive feedback from coparticipants and facilitators.
Haploinsufficiency of USP7, located at chromosome 16p13.2, has recently been reported in seven individuals with neurodevelopmental phenotypes, including developmental delay/intellectual disability ...(DD/ID), autism spectrum disorder (ASD), seizures, and hypogonadism. Further, USP7 was identified to critically incorporate into the MAGEL2-USP7-TRIM27 (MUST), such that pathogenic variants in USP7 lead to altered endosomal F-actin polymerization and dysregulated protein recycling.
We report 16 newly identified individuals with heterozygous USP7 variants, identified by genome or exome sequencing or by chromosome microarray analysis. Clinical features were evaluated by review of medical records. Additional clinical information was obtained on the seven previously reported individuals to fully elucidate the phenotypic expression associated with USP7 haploinsufficiency.
The clinical manifestations of these 23 individuals suggest a syndrome characterized by DD/ID, hypotonia, eye anomalies,feeding difficulties, GERD, behavioral anomalies, and ASD, and more specific phenotypes of speech delays including a nonverbal phenotype and abnormal brain magnetic resonance image findings including white matter changes based on neuroradiologic examination.
The consistency of clinical features among all individuals presented regardless of de novo USP7 variant type supports haploinsufficiency as a mechanism for pathogenesis and refines the clinical impact faced by affected individuals and caregivers.
Although much is known about signs, symptoms, and management in the acute phase of crotaline snake envenomation, little is known about signs, symptoms, function, and quality of life during the ...recovery phase. The purpose of this observational pilot investigation is to evaluate the utility of several clinical outcome instruments in the setting of copperhead snakebite, and to characterize the clinical course of recovery.
This is a multi-center prospective, open-label, observational study of patients envenomated by copperhead snakes. We administered the Disabilities of the Arm, Shoulder, and Hand (DASH), Lower Extremity Functional Scale (LEFS), Patient-Specific Functional Scale (PSFS), Work Productivity and Ability Impairment: Special Health Problem (WPAI: SHP), Patients' Global Impression of Change (PGIC), Patient's Global Assessment of Recovery (PGAR), and SF-36 instruments, obtained numeric pain rating scales, and measured grip strength, walking speed, and swelling prior to hospital discharge and 3, 7, 14, 21, and 28 days after envenomation.
20 subjects were enrolled; none were lost to follow-up. Most (80%) had moderate severity swelling, and most (75%) received antivenom. Across the broad range of measures, abnormalities of pain, swelling, impairments of physical and role function, and quality of life persisted for 7-14 days in most subjects. Validated self-reported outcome measures, such as the DASH, LEFS, PSFS, PGIC, SF-36, and the daily activities impairment portion of the WPAI: SHP were more responsive than measurements of swelling or walking speed. Data quality issues limited the utility of the work impairment portion of the WPAI: SHP. Residual signs, symptoms, and impairment in some subjects lasted through the 28-day study period. The study design precluded any assessment of the effectiveness of antivenom.
Signs, symptoms, impaired function, and decreased quality of life typically last 7 - 14 days after copperhead envenomation. Several tools appear responsive and useful in studying recovery from pit viper envenomation.
ClinicalTrials.gov NCT01651299.
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