Mice Lacking the Circadian Modulators SHARP1 and SHARP2 Display Altered Sleep and Mixed State Endophenotypes of Psychiatric Disorders

Increasing evidence suggests that clock genes may be implicated in a spectrum of psychiatric diseases, including sleep and mood related disorders as well as schizophrenia. The bHLH transcription factors SHARP1/DEC2/BHLHE41 and SHARP2/DEC1/ BHLHE40 are modulators of the circadian system and SHARP1/DEC2/BHLHE40 has been shown to regulate homeostatic sleep drive in humans. In this study, we characterized Sharp1 and Sharp2 double mutant mice (S1/2(-/-)) using online EEG recordings in living animals, behavioral assays and global gene expression profiling. EEG recordings revealed attenuated sleep/wake amplitudes and alterations of theta oscillations. Increased sleep in the dark phase is paralleled by reduced voluntary activity and cortical gene expression signatures reveal associations with psychiatric diseases. S1/2(-/-) mice display alterations in novelty induced activity, anxiety and curiosity. Moreover, mutant mice exhibit impaired working memory and deficits in prepulse inhibition resembling symptoms of psychiatric diseases. Network modeling indicates a connection between neural plasticity and clock genes, particularly for SHARP1 and PER1. Our findings support the hypothesis that abnormal sleep and certain (endo) phenotypes of psychiatric diseases may be caused by common mechanisms involving components of the molecular clock including SHARP1 and SHARP2

Artificial-intelligence-based molecular classification of diffuse gliomas using rapid, label-free optical imaging

Molecular classification has transformed the management of brain tumors by enabling more accurate prognostication and personalized treatment. However, timely molecular diagnostic testing for patients with brain tumors is limited, complicating surgical and adjuvant treatment and obstructing clinical trial enrollment. In this study, we developed DeepGlioma, a rapid ($< 90$ seconds), artificial-intelligence-based diagnostic screening system to streamline the molecular diagnosis of diffuse gliomas. DeepGlioma is trained using a multimodal dataset that includes stimulated Raman histology (SRH); a rapid, label-free, non-consumptive, optical imaging method; and large-scale, public genomic data. In a prospective, multicenter, international testing cohort of patients with diffuse glioma ($n=153$) who underwent real-time SRH imaging, we demonstrate that DeepGlioma can predict the molecular alterations used by the World Health Organization to define the adult-type diffuse glioma taxonomy (IDH mutation, 1p19q co-deletion and ATRX mutation), achieving a mean molecular classification accuracy of $93.3\pm 1.6\%$. Our results represent how artificial intelligence and optical histology can be used to provide a rapid and scalable adjunct to wet lab methods for the molecular screening of patients with diffuse glioma.Comment: Paper published in Nature Medicin

Disturbed Clockwork Resetting in Sharp-1 and Sharp-2 Single and Double Mutant Mice

BACKGROUND: The circadian system provides the basis to anticipate and cope with daily recurrent challenges to maintain the organisms' homeostasis. De-synchronization of circadian feedback oscillators in humans causes 'jet lag', likely contributes to sleep-, psychiatric-, metabolic disorders and even cancer. However, the molecular mechanisms leading to the disintegration of tissue-specific clocks are complex and not well understood. METHODOLOGY/PRINCIPAL FINDINGS: Based on their circadian expression and cell culture experiments, the basic Helix-Loop-Helix (bHLH) transcription factors SHARP-1(Dec2) and SHARP-2(Stra13/Dec1) were proposed as novel negative regulators of the molecular clock. To address their function in vivo, we generated Sharp-1 and Sharp-2 single and double mutant mice. Our experiments reveal critical roles for both factors in regulating period length, tissue-specific control of clock gene expression and entrainment to external cues. Light-pulse experiments and rapid delays of the light-dark cycle (experimental jet lag) unravel complementary functions for SHARP-1 and SHARP-2 in controlling activity phase resetting kinetics. Moreover, we show that SHARP-1 and 2 can serve dual functions as repressors and co-activators of mammalian clock gene expression in a context-specific manner. This correlates with increased amplitudes of Per2 expression in the cortex and liver and a decrease in the suprachiasmatic nucleus (SCN) of double mutant mice. CONCLUSIONS/SIGNIFICANCE: The existence of separate mechanisms regulating phase of entrainment, rhythm amplitude and period length has been postulated before. The differential effects of Sharp-deficiency on rhythmicity and behavioral re-entrainment, coupled to tissue-dependent regulatory functions, provide a new mechanistic basis to further understand the complex process of clock synchronizations

Eleven strategies for making reproducible research and open science training the norm at research institutions

Across disciplines, researchers increasingly recognize that open science and reproducible research practices may accelerate scientific progress by allowing others to reuse research outputs and by promoting rigorous research that is more likely to yield trustworthy results. While initiatives, training programs, and funder policies encourage researchers to adopt reproducible research and open science practices, these practices are uncommon inmanyfields. Researchers need training to integrate these practicesinto their daily work. We organized a virtual brainstorming event, in collaboration with the German Reproducibility Network, to discuss strategies for making reproducible research and open science training the norm at research institutions. Here, weoutline eleven strategies, concentrated in three areas:(1)offering training, (2)adapting research assessment criteria and program requirements, and (3) building communities. We provide a brief overview of each strategy, offer tips for implementation,and provide links to resources. Our goal is toencourage members of the research community to think creatively about the many ways they can contribute and collaborate to build communities,and make reproducible research and open sciencetraining the norm. Researchers may act in their roles as scientists, supervisors, mentors, instructors, and members of curriculum, hiring or evaluation committees. Institutionalleadership and research administration andsupport staff can accelerate progress by implementing change across their institution

Eleven strategies for making reproducible research and open science training the norm at research institutions

Across disciplines, researchers increasingly recognize that open science and reproducible research practices may accelerate scientific progress by allowing others to reuse research outputs and by promoting rigorous research that is more likely to yield trustworthy results. While initiatives, training programs, and funder policies encourage researchers to adopt reproducible research and open science practices, these practices are uncommon inmanyfields. Researchers need training to integrate these practicesinto their daily work. We organized a virtual brainstorming event, in collaboration with the German Reproducibility Network, to discuss strategies for making reproducible research and open science training the norm at research institutions. Here, weoutline eleven strategies, concentrated in three areas:(1)offering training, (2)adapting research assessment criteria and program requirements, and (3) building communities. We provide a brief overview of each strategy, offer tips for implementation,and provide links to resources. Our goal is toencourage members of the research community to think creatively about the many ways they can contribute and collaborate to build communities,and make reproducible research and open sciencetraining the norm. Researchers may act in their roles as scientists, supervisors, mentors, instructors, and members of curriculum, hiring or evaluation committees. Institutionalleadership and research administration andsupport staff can accelerate progress by implementing change across their institution

A View from the Past Into our Collective Future: The Oncofertility Consortium Vision Statement

Today, male and female adult and pediatric cancer patients, individuals transitioning between gender identities, and other individuals facing health extending but fertility limiting treatments can look forward to a fertile future. This is, in part, due to the work of members associated with the Oncofertility Consortium. The Oncofertility Consortium is an international, interdisciplinary initiative originally designed to explore the urgent unmet need associated with the reproductive future of cancer survivors. As the strategies for fertility management were invented, developed or applied, the individuals for who the program offered hope, similarly expanded. As a community of practice, Consortium participants share information in an open and rapid manner to addresses the complex health care and quality-of-life issues of cancer, transgender and other patients. To ensure that the organization remains contemporary to the needs of the community, the field designed a fully inclusive mechanism for strategic planning and here present the findings of this process. This interprofessional network of medical specialists, scientists, and scholars in the law, medical ethics, religious studies and other disciplines associated with human interventions, explore the relationships between health, disease, survivorship, treatment, gender and reproductive longevity. The goals are to continually integrate the best science in the service of the needs of patients and build a community of care that is ready for the challenges of the field in the future

Cognitive Vulnerability in the Context of Panic: Assessment of Panic-Related Associations and Interpretations in Individuals with Varying Levels of Anxiety Sensitivity

Background: Cognitive models of panic disorder (PD) highlight the role of panic-related associations and interpretations. However, results are mixed and rely on specific measures. This study examined panic-related associations and interpretations using established and new paradigms in individuals varying on anxiety sensitivity (AS). Methods: Associations were measured using a priming task and a novel Single Target Implicit Association Test (STIAT); interpretations were assessed using the Interpretation Bias Questionnaire (IBQ) and a novel Scrambled Sentences Task (SST). Symptoms were assessed via a provocation task (Straw Breathing Task, SBT). Results: Panic-related interpretations correlated with AS and other PD-related measures. Of the association tasks, only the priming task correlated with one of the other PD-related measures. Panic-related interpretations assessed via the SST, but not priming, STIAT, and IBQ, predicted SBT reactivity. The relationship between AS and SBT reactivity was mediated by panic-related interpretations. Conclusions Our data provide support for panic-related interpretations as an important cognitive mechanism. (c) Springer Science+Business Medi

Analysis of transient phosphorylation-dependent protein-protein interactions in living mammalian cells using split-TEV-7

Hosphorylated resulting in binding domains for several adapter proteins such as the PI3Kp85 subunits. In the split-TEV assays, ErbB4 is fused to N-TEV, a TEV cleavage site and the transcription factor GV (ErbB4-N-TEV-GV). The adapter proteins are fused to C-TEV (such as PI3Kp85-C-TEV). PI3Kp85 binding to ErbB4 results in the formation of a functional TEV protease made up of the N-TEV and C-TEV fragments. Proteolytic activity releases GV, which can translocate to the nucleus to induce transcription of a luciferase reporter gene. (b) Schematic drawing of the experimental setup for the 2-cell-population assay to allow ErbB4 activation exclusively at the cell membrane. The ErbB4 receptor (dark green rectangle) and the adapters (bright green circle) were transfected into population 1. The ligand neuregulin (Nrg) was transfected into population 2. This verifies that the Nrg induced ErbB4 activation and the subsequent phosphorylation-dependent interaction with the adapter protein are initiated via intercellular signaling at the cell membrane. (c) Visualisation of the 2-population assay. EYFP was co-transfected into population 1, ECFP into population 2. EYFP and ECFP were allowed to express for 20 h separately. Subsequently, both populations were mixed, and the assay was continued for additional 24 h. The merged image shows that YFP and CFP signals are not overlapping and that yellow and cyan signals are frequently in close proximity. Images were taken immediately before the luciferase assay was performed. (d) 2-population luciferase assay depicting the activation of the neuregulin-1 (Nrg1-II-b1a variant) induced phosphorylation-dependent interaction between the ErbB4 receptor and the adapter proteins PI3Kp85α, PI3Kp85β, Grb2 and Shc1. Combination of constructs were transfected into PC12 cells as indicated. RLUs, relative luciferase units; n = 6. (e) Renilla luciferase readings taken from the same assay as depicted in (d). Stimulation with Nrg1-II-b1a does not lead to substantial differences in absolute Renilla luciferase readings.<p><b>Copyright information:</b></p><p>Taken from "Analysis of transient phosphorylation-dependent protein-protein interactions in living mammalian cells using split-TEV"</p><p>http://www.biomedcentral.com/1472-6750/8/55</p><p>BMC Biotechnology 2008;8():55-55.</p><p>Published online 13 Jul 2008</p><p>PMCID:PMC2483975.</p><p></p