Immunological isolation and characterization of neuronal progenitors from human dental pulp: A laboratory-based investigation.

AIMS: Dental pulp stem cells (DPSCs) contain a population of stem cells with a broad range of differentiation potentials, as well as more lineage-committed progenitors. Such heterogeneity is a significant obstacle to experimental and clinical applications. The aim of this study is to isolate and characterize a homogenous neuronal progenitor cell population from human DPSCs. METHODOLOGY: Polysialylated-neural cell adhesion molecule (PSA-NCAM+) neural progenitors were isolated from the dental pulp of three independent donors using magnetic-activated cell sorting (MACS) technology. Immunofluorescent staining with a panel of neural and non-neural markers was used to characterize the magnetically isolated PSA-NCAM+ fraction. PSA-NCAM+ cells were then cultured in Neurobasal A supplemented with neurotrophic factors: dibutyryl cyclic-AMP, neurotrophin-3, B27 and N2 supplements to induce neuronal differentiation. Both PSA-NCAM+ and differentiated PSA-NCAM+ cells were used in Ca2+ imaging studies to assess the functionality of P2X3 receptors as well as membrane depolarization. RESULTS: PSA-NCAM+ neural progenitors were isolated from a heterogeneous population of hDPSCs using magnetic-activated cell sorting and anti-PSA-NCAM MicroBeads. Flow cytometry analysis demonstrated that immunomagnetic sorting significantly increased the purity of PSA-NCAM+ cells. Immunofluorescent staining revealed expression of pan-neuronal and mature neuronal markers, PGP9.5 and MAP2, respectively, as well as weak expression of the mature sensory markers, peripherin and islet1. ATP-induced response was mediated predominately by P2X3 receptors in both undifferentiated and differentiated cells, with a greater magnitude observed in the latter. In addition, membrane depolarizations were also detected in cells before and after differentiation when loaded with fast-voltage-responding fluorescent molecule, FluoVolt™ in response to potassium chloride. Interestingly, only differentiated PSA-NCAM+ cells were capable of spontaneous membrane oscillations. CONCLUSIONS: In summary, DPSCs contain a population of neuronal progenitors with enhanced neural differentiation and functional neural-like properties that can be effectively isolated with magnetic-activated cell sorting (MACS).

Post-ischemic reorganization of sensory responses in cerebral cortex.

Introduction: Sensorimotor integration is critical for generating skilled, volitional movements. While stroke tends to impact motor function, there are also often associated sensory deficits that contribute to overall behavioral deficits. Because many of the cortico-cortical projections participating in the generation of volitional movement either target or pass-through primary motor cortex (in rats, caudal forelimb area; CFA), any damage to CFA can lead to a subsequent disruption in information flow. As a result, the loss of sensory feedback is thought to contribute to motor dysfunction even when sensory areas are spared from injury. Previous research has suggested that the restoration of sensorimotor integration through reorganization or de novo neuronal connections is important for restoring function. Our goal was to determine if there was crosstalk between sensorimotor cortical areas with recovery from a primary motor cortex injury. First, we investigated if peripheral sensory stimulation would evoke responses in the rostral forelimb area (RFA), a rodent homologue to premotor cortex. We then sought to identify whether intracortical microstimulation-evoked activity in RFA would reciprocally modify the sensory response. Methods: We used seven rats with an ischemic lesion of CFA. Four weeks after injury, the rats' forepaw was mechanically stimulated under anesthesia and neural activity was recorded in the cortex. In a subset of trials, a small intracortical stimulation pulse was delivered in RFA either individually or paired with peripheral sensory stimulation. Results: Our results point to post-ischemic connectivity between premotor and sensory cortex that may be related to functional recovery. Premotor recruitment during the sensory response was seen with a peak in spiking within RFA after the peripheral solenoid stimulation despite the damage to CFA. Furthermore, stimulation in RFA modulated and disrupted the sensory response in sensory cortex. Discussion: The presence of a sensory response in RFA and the sensitivity of S1 to modulation by intracortical stimulation provides additional evidence for functional connectivity between premotor and somatosensory cortex. The strength of the modulatory effect may be related to the extent of the injury and the subsequent reshaping of cortical connections in response to network disruption.

Investigating gender differences in the co-occurrence of PTSD and food addiction.

OBJECTIVE: Post-traumatic stress disorder (PTSD) commonly co-occurs with substance use disorders (SUDs). Past studies suggest PTSD is also associated with food addiction (compulsive intake of highly processed foods containing refined carbohydrates and/or added fat). However, research investigating gender differences has been limited (e.g., restricted samples) and mixed. We aim to investigate the risk of co-occurring PTSD and food addiction in a community sample for all participants and stratified by gender. Additionally, we conducted risk ratios for problematic substance use and obesity to allow for within-sample comparisons. METHOD: We utilized a sample of 318 participants recruited from Amazon Mechanical Turk (mean age = 41.2, 47.8% men, 78.0% white) to address existing gaps in the literature on PTSD and food addiction. We calculated risk ratios (adjusted for sociodemographic covariates) using modified Poisson regression with 95% confidence intervals. Results were also gender stratified. RESULTS: Risk of food addiction (Risk Ratio (RR) = 6.42, 95% CI [4.10, 10.07], problematic alcohol use (RR) = 3.86, 95% CI [2.25,6.62], problematic smoking (RR) = 3.93, 95% CI [2.22, 6.97], and problematic nicotine vaping (RR) = 5.41, 95% CI [2.41, 11.14] were higher for those meeting criteria for PTSD. Risk of problematic cannabis use, and risk of obesity were not significantly higher for those meeting criteria for PTSD. Gender-stratified results suggest risk of food addiction may be higher for men (RR) = 8.54, 95% CI [4.49, 16.25] compared to women (RR) = 4.32, 95% CI [2.16, 8.62]. DISCUSSION: Food addiction, but not obesity, appears to co-occur with PTSD more strongly than other types of problematic substance use (alcohol, cannabis, cigarettes, nicotine vaping). This risk appears to be particularly high for men compared to women. Assessing for food addiction in those with PTSD, particularly in men, may assist in identifying high-risk groups.

A Tool to Assess Competence in Critical Care Ultrasound Based on Entrustable Professional Activities.

Background: Existing assessment tools for competence in critical care ultrasound (CCUS) have limited scope and interrupt clinical workflow. The framework of entrustable professional activities (EPAs) is well suited to developing an assessment tool that is comprehensive and readily integrated into the intensive care unit (ICU) training environment. Objective: This study sought to design an EPA-based tool to assess competence in CCUS for pulmonary and critical care fellows and to assess the validity and reliability of the tool. Methods: Eight experts in CCUS met to define the core EPAs for CCUS. A nominal group technique was used to reach consensus. An assessment tool was created based on the EPAs with a modified Ottawa entrustability scale. Trained faculty evaluated pulmonary and critical care fellows using this tool in the ICU over a 6-month study period at a single institution. An assessment of validity of the EPA-based tool is made with four sources of validity evidence: content, response process, reliability, and relation to other variables. Reliability and response process data were generated using generalizability theory analysis to estimate sources of variance in entrustment scores. Analysis of response process validity and validity by relation to other variables was performed using regression models. Results: Fifty-four assessments were recorded during the study period, conducted on 23 trainees by 13 faculty. Content validity of the tool was demonstrated using expert consensus and published guidelines from critical care societies to define the EPAs. Response process validity was demonstrated by the low variance in entrustment scores due to evaluators (0.086 or 6%) and high agreement between score and trainee self-assessment (regression coefficient, 0.82; P < 0.0001). Reliability was demonstrated by the high "true" variance in entrustment score attributable to the trainee: 0.674 or 45%. Validity by relation to other variables was demonstrated using regression analysis to show correlation between entrustment score and the number of times a fellow has performed an EPA (regression coefficient, 0.023; P < 0.0001). Conclusion: An EPA-based assessment tool for competence in CCUS was created. We obtained sufficient validity evidence on three of the diagnostic EPAs. Procedural EPAs were infrequently assessed, limiting generalizability in this subgroup.

Development and Validation of a Novel Instrument to Measure the Community Well-Being of Residency Programs.

PURPOSE: To develop and validate the Residency Community Well-Being (RCWB) instrument, a novel instrument to measure the subjective community well-being of an individual residency program, and to explore differences in RCWB scores between demographic groups. METHOD: An initial questionnaire to measure a residency program's community well-being was developed after literature review. Items were pilot tested, and the questionnaire was reviewed by experts in the fields of residency education, survey design, and sociology. The questionnaire was administered electronically between March and July 2021 to U.S. residents in 18 specialties recruited through convenience and snowball sampling using social media, a listserv, and personal emails to residency program leaders. Three previously validated instruments were administered as well to examine criterion validity: the Professional Fulfillment Index, the Brief Inventory of Thriving, and a single-item burnout measure. Data were analyzed with descriptive statistics, and exploratory factor analysis was performed using principal axis factoring with direct oblimin rotation to reduce the items and identify subscales. RESULTS: Of the 366 participants who opened and started the survey, 219 completed it (completion rate: 59.8%). Most respondents were women (133, 60.7%), 26-30 years old (132, 60.3%), and White (149, 68.0%). Three subscales emerged with 18 items: program leadership, structures, and practices (PLSP); resident interpersonal relationships (RIR); and resident mistreatment (RM). The Cronbach's alphas were 0.96 for PLSP, 0.92 for RIR, 0.82 for RM, and 0.95 for the overall RCWB. RCWB score positively correlated with professional fulfillment ( r = .52, P < .001) and thriving ( r = .45, P < .001) and inversely correlated with burnout ( r = -.39, P < .001). CONCLUSIONS: The RCWB instrument demonstrates strong internal consistency and content and criterion validity that shows that a residency program's subjective community well-being is primarily composed of program leadership quality, supportive interpersonal relationships, and the absence of mistreatment.

Post-Ischemic Reorganization of Sensory Responses in Cerebral Cortex.

Sensorimotor integration is critical for generating skilled, volitional movements. While stroke tends to impact motor function, there are also often associated sensory deficits that contribute to overall behavioral deficits. Because many of the cortico-cortical projections participating in the generation of volitional movement either target or pass-through primary motor cortex (in rats, caudal forelimb area; CFA), any damage to CFA can lead to a subsequent disruption in information flow. As a result, the loss of sensory feedback is thought to contribute to motor dysfunction even when sensory areas are spared from injury. Previous research has suggested that the restoration of sensorimotor integration through reorganization or de novo neuronal connections is important for restoring function. Our goal was to determine if there was crosstalk between sensorimotor cortical areas with recovery from a primary motor cortex injury. First, we investigated if peripheral sensory stimulation would evoke responses in the rostral forelimb area (RFA), a rodent homologue to premotor cortex. We then sought to identify whether intracortical microstimulation-evoked activity in RFA would reciprocally modify the sensory response. We used seven rats with an ischemic lesion of CFA. Four weeks after injury, the rats' forepaw was mechanically stimulated under anesthesia and neural activity was recorded in the cortex. In a subset of trials, a small intracortical stimulation pulse was delivered in RFA either individually or paired with peripheral sensory stimulation. Our results point to post-ischemic connectivity between premotor and sensory cortex that may be related to functional recovery. Premotor recruitment during the sensory response was seen with a peak in spiking within RFA after the peripheral solenoid stimulation despite the damage to CFA. Furthermore, stimulation evoked activity in RFA modulated and disrupted the sensory response in sensory cortex, providing additional evidence for the transmission of premotor activity to sensory cortex and the sensitivity of sensory cortex to premotor cortex's influence. The strength of the modulatory effect may be related to the extent of the injury and the subsequent reshaping of cortical connections in response to network disruption.

Focusing on the Positive: Home Health Aides' Desire for Autonomy and Control.

Home Health Aides (HHAs) are one of the fastest growing workforces in the country, yet the industry struggles to recruit and retain workers. This study explored HHAs' experiences with the level of control, autonomy, and decision-making authority in their work. Six focus groups with 37 HHAs were conducted in Massachusetts. Findings showed that HHAs viewed control as a positive job characteristic, which attracted them to and led them to remain in the position. Positive benefits included having control over client selection, location, hours, and the ability to determine their day-to-day tasks and schedules. The study results highlight the value that HHAs place on autonomy and control and the potential benefit that these job qualities have for greater recruitment and retention of these workers. Amplifying control by bolstering training and expanding scope of practice may entice new individuals to pursue a HHA career and help maintain those currently in the position.

Co-occurrence of food addiction, obesity, problematic substance use, and parental history of problematic alcohol use.

OBJECTIVE: The present study investigates the rates of co-occurrence among food addiction (FA), problematic substance use (alcohol, cannabis, cigarettes, nicotine vaping), parental history of problematic alcohol use, and obesity as an important step to understanding whether an addictive-like eating phenotype exists. METHOD: A community sample of 357 U.S. adults (49.7% male, 77.6% White, Mage 40.7) completed the Yale Food Addiction Scale 2.0 (YFAS2.0), the Alcohol Use Disorders Identification Test, the Cannabis Use Disorders Identification Test, the Fagerstrom Test for Nicotine Dependence, the E-Cigarette Dependence Scale, the Family Tree Questionnaire, and demographic/self-report body mass index questions through Amazon Mechanical Turk. Risk ratios (RRs; unadjusted and adjusted for sociodemographic covariates) were calculated using modified Poisson's regression. RESULTS: Risk of FA was higher in participants with problematic alcohol use (RR = 2.13, 99% CI [1.32, 3.45]), smoking (RR = 1.86, 99% CI [0.82, 3.36]), cannabis use (unadjusted; RR = 2.22, 99% CI [1.17, 4.18]), vaping (RR = 2.71, 99% CI [1.75, 4.21]), and parental history of problematic alcohol use (RR = 2.35, 99% CI [1.46, 3.79]). Risk of FA in participants with obesity was only higher in adjusted models (RR = 1.87, 99% CI [1.06, 3.27]). Obesity was not significantly associated with problematic substance use and parental history of problematic alcohol use. CONCLUSIONS: FA, but not obesity, co-occurred with problematic substance use and a parental history of problematic alcohol use. Results support the conceptualization of FA as an addictive disorder. The inclusion of FA as an addictive disorder in diagnostic frameworks is an important area of future consideration. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Temperature-Dependent Constrained Diffusion of Micro-Confined Alkylimidazolium Chloride Ionic Liquids.

Alkylimidazolium chloride ionic liquids (ILs) have many uses in a variety of separation systems, including micro-confined separation systems. To understand the separation mechanism in these systems, the diffusion properties of analytes in ILs under relevant operating conditions, including micro-confinement dimension and temperature, should be known. For example, separation efficiencies for various IL-based microextraction techniques are dependent on the sample volume and temperature. Temperature-dependent (20-100 °C) fluorescence recovery after photobleaching (FRAP) was utilized to determine the diffusion properties of a zwitterionic, hydrophilic dye, ATTO 647, in alkylimidazolium chloride ILs in micro-confined geometries. These micro-confined geometries were generated by sandwiching the IL between glass substrates that were separated by ∼1 to 100 µm. From the measured temperature-dependent FRAP data, we note alkyl chain length-, thickness-, and temperature-dependent diffusion coefficients, with values ranging from 0.021 to 46 µm2/s. Deviations from Brownian diffusion are observed at lower temperatures and increasingly less so at elevated temperatures; the differences are attributed to alterations in intermolecular interactions that reduce temperature-dependent nanoscale structural heterogeneities. The temperature- and thickness-dependent data provide a useful foundation for efficient design of micro-confined IL separation systems.

Childhood trauma and food addiction: The role of emotion regulation difficulties and gender differences.

OBJECTIVE: Childhood trauma has been associated with substance use disorders (SUDs), but less research has investigated its association with food addiction (i.e., compulsive intake of highly processed foods containing refined carbohydrates and/or added fat). Existing research on childhood trauma and food addiction has focused primarily on women from restricted samples. Further, emotion dysregulation difficulties are implicated in both childhood trauma and food addiction, but research has not explored whether it may play a mediating role. METHOD: The current study utilizes a sample of 310 participants recruited from Amazon Mechanical Turk (mean age = 41.3 years old, 47.4% men, 78.7% white) to investigate the mediating role of emotion dysregulation in the association between childhood trauma and food addiction. We also conducted exploratory analyses to investigate for gender differences in these associations. Gender-stratified correlational matrixes were conducted to investigate associations between childhood trauma, food addiction, and emotional dysregulation. Moderated mediation models and multilevel regressions were also conducted to identify the role of gender in the association between childhood trauma, food addiction, and emotion dysregulation. RESULTS: Emotion dysregulation was found to partially mediate the associations between food addiction and childhood trauma and gender was found to moderate associations between childhood trauma and emotion dysregulation, as well as childhood trauma and food addiction. Both moderating pathways were significantly stronger for men compared to women. DISCUSSION: Results suggests that emotion dysregulation may be one important mediator in the association between childhood trauma and food addiction, particularly for men. The identification of other potential mechanisms contributing to the association between childhood trauma and food addiction and the use of longitudinal measurement strategies will be important in future research.

Yoga therapy knowledge in third-year pharmacy students: An education intervention.

BACKGROUND AND PURPOSE: The omission of yoga education in health professions curricula limits health professionals from understanding how to support patient well-being in an integrated manner. The objective of this study was to examine the effectiveness of a yoga therapy-based educational intervention in increasing yoga knowledge of third-year pharmacy students. EDUCATIONAL ACTIVITY AND SETTING: Students in the Pharmacognosy and Complementary and Alternative Medicine course received a three-hour educational intervention over two class meetings on yoga therapy during fall 2019. The pedagogical method of peer instruction was used, and pre- and post-questionnaires were administered online prior to and after the intervention. The instrument contained eight demographic, six yoga knowledge and participation, 10 yoga therapy, and 15 referral behavior questions. FINDINGS: A statistically significant increase in yoga knowledge scores from 1.43 ± 0.9 before the education intervention to 5.69 ± 1.65 after the intervention occurred (P < .01) was found. A majority believed that yoga therapy should be included in the curriculum. SUMMARY: Increases in post-questionnaire knowledge scores suggested that the educational intervention increased pharmacy students' knowledge of yoga therapy in our sample. Studies specific to yoga education in health professional students, especially in the United States, are scarce. Additionally, the impact of an educational intervention on yoga knowledge in pharmacy students has not been studied. Including yoga therapy in pharmacy curricula should be explored as greater integration of yoga in healthcare is likely to occur due to its effectiveness as an adjunct and widespread use.

Maximizing Home Health Aide Retention: The Impact of Control and Support on the Job.

BACKGROUND AND OBJECTIVES: This study examined relationships between the level of control and support and home health aides (HHAs) job satisfaction and intent to leave the job. RESEARCH DESIGN AND METHODS: Data derive from a survey of 512 HHAs in Massachusetts. Logistic regression using generalized estimating equations was employed for the analysis. Dependent variables included satisfaction and intent to leave the job as a home care aide generally and satisfaction and intent to leave the job at the aide's current agency. RESULTS: The findings showed that greater control and support on the job were important predictors of positive work outcomes, controlling for job demands and other covariates. The odds of HHAs being satisfied with their job as a home care aide increased with the degree of control, whereas the odds of HHAs being satisfied with their job at their current agency increased with the extent of support. Control was negatively associated with HHAs' intent to leave the job as an aide; no relationship was found between control or support and HHAs' intent to leave their current agency. DISCUSSION AND IMPLICATIONS: The results from this study illustrate the importance for HHAs of having control and autonomy in their work, as well as the benefit of support from supervisors and the home care agency, on satisfaction and intent to leave. Expanding HHA's ability to maintain control over their day-to-day work, as well as enhancing the supports available to them, is likely to benefit home care workers, clients, and agencies through increased retention.

The CHD4-related syndrome: a comprehensive investigation of the clinical spectrum, genotype-phenotype correlations, and molecular basis.

PURPOSE: Sifrim-Hitz-Weiss syndrome (SIHIWES) is a recently described multisystemic neurodevelopmental disorder caused by de novo variants inCHD4. In this study, we investigated the clinical spectrum of the disorder, genotype-phenotype correlations, and the effect of different missense variants on CHD4 function. METHODS: We collected clinical and molecular data from 32 individuals with mostly de novo variants in CHD4, identified through next-generation sequencing. We performed adenosine triphosphate (ATP) hydrolysis and nucleosome remodeling assays on variants from five different CHD4 domains. RESULTS: The majority of participants had global developmental delay, mild to moderate intellectual disability, brain anomalies, congenital heart defects, and dysmorphic features. Macrocephaly was a frequent but not universal finding. Additional common abnormalities included hypogonadism in males, skeletal and limb anomalies, hearing impairment, and ophthalmic abnormalities. The majority of variants were nontruncating and affected the SNF2-like region of the protein. We did not identify genotype-phenotype correlations based on the type or location of variants. Alterations in ATP hydrolysis and chromatin remodeling activities were observed in variants from different domains. CONCLUSION: The CHD4-related syndrome is a multisystemic neurodevelopmental disorder. Missense substitutions in different protein domains alter CHD4 function in a variant-specific manner, but result in a similar phenotype in humans.

Correction: The CHD4-related syndrome: a comprehensive investigation of the clinical spectrum, genotype-phenotype correlations, and molecular basis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Immediate aftermath of a client's death: the experience of home health aides.

This study investigated Home Health Aides' (HHAs) experiences in the immediate aftermath of client death. Semi-structured in-person interviews with 80 HHAs explored how notification of death and reassignment to a new client were handled. Only 42.5% of HHAs were notified of the death; 40% had to notify the agency; 17.5% were not notified at all and had a negative experience. Reassignment preferences varied, but HHAs had a better experience when their preferences were taken into consideration. Study findings suggest that more mindful approaches to transitions following client death would be valued by HHAs and could improve their work experience.

A Post-Traumatic Stress Disorder review: the prevalence of underreporting and the role of stigma in the Military.

Exposure to traumatic events has been a part of human existence since the beginning of time; however modern lifestyles and developments mean that the majority of the population are spared the affliction of extreme trauma. The military, however, are one subgroup of individuals who are actively and repeatedly exposed to terrifying events on a regular basis, putting them at risk of developing "Post-Traumatic Stress Disorder" (PTSD). Despite this, the prevalence of PTSD reports in the UK military remains lower than its international allies, suggesting this value may be an underestimation of the true prevalence. Wider investigation of the potential causes of this has highlighted the stigmatization of mental illness in the military as the key barrier to the help-seeking behaviours. However, the effect of national de-stigmatization programs on PTSD help seeking is unclear. This review aims to determine the prevalence of PTSD within the UK military in the context of other international powers and examine how stigmatization of mental illness in military may contribute to this. The international value of de-stigmatization programs will be debated and future directions for research suggested.

From bedside to bench and back: Translating ASD models.

Autism spectrum disorders (ASD) represent a heterogeneous group of disorders defined by deficits in social interaction/communication and restricted interests, behaviors, or activities. Models of ASD, developed based on clinical data and observations, are used in basic science, the "bench," to better understand the pathophysiology of ASD and provide therapeutic options for patients in the clinic, the "bedside." Translational medicine creates a bridge between the bench and bedside that allows for clinical and basic science discoveries to challenge one another to improve the opportunities to bring novel therapies to patients. From the clinical side, biomarker work is expanding our understanding of possible mechanisms of ASD through measures of behavior, genetics, imaging modalities, and serum markers. These biomarkers could help to subclassify patients with ASD in order to better target treatments to a more homogeneous groups of patients most likely to respond to a candidate therapy. In turn, basic science has been responding to developments in clinical evaluation by improving bench models to mechanistically and phenotypically recapitulate the ASD phenotypes observed in clinic. While genetic models are identifying novel therapeutics targets at the bench, the clinical efforts are making progress by defining better outcome measures that are most representative of meaningful patient responses. In this review, we discuss some of these challenges in translational research in ASD and strategies for the bench and bedside to bridge the gap to achieve better benefits to patients.

Impact of Variable RNA-Sequencing Depth on Gene Expression Signatures and Target Compound Robustness: Case Study Examining Brain Tumor (Glioma) Disease Progression.

PURPOSE: Gene expression profiling can uncover biologic mechanisms underlying disease and is important in drug development. RNA sequencing (RNA-seq) is routinely used to assess gene expression, but costs remain high. Sample multiplexing reduces RNAseq costs; however, multiplexed samples have lower cDNA sequencing depth, which can hinder accurate differential gene expression detection. The impact of sequencing depth alteration on RNA-seq-based downstream analyses such as gene expression connectivity mapping is not known, where this method is used to identify potential therapeutic compounds for repurposing. METHODS: In this study, published RNA-seq profiles from patients with brain tumor (glioma) were assembled into two disease progression gene signature contrasts for astrocytoma. Available treatments for glioma have limited effectiveness, rendering this a disease of poor clinical outcome. Gene signatures were subsampled to simulate sequencing alterations and analyzed in connectivity mapping to investigate target compound robustness. RESULTS: Data loss to gene signatures led to the loss, gain, and consistent identification of significant connections. The most accurate gene signature contrast with consistent patient gene expression profiles was more resilient to data loss and identified robust target compounds. Target compounds lost included candidate compounds of potential clinical utility in glioma (eg, suramin, dasatinib). Lost connections may have been linked to low-abundance genes in the gene signature that closely characterized the disease phenotype. Consistently identified connections may have been related to highly expressed abundant genes that were ever-present in gene signatures, despite data reductions. Potential noise surrounding findings included false-positive connections that were gained as a result of gene signature modification with data loss. CONCLUSION: Findings highlight the necessity for gene signature accuracy for connectivity mapping, which should improve the clinical utility of future target compound discoveries.

A New Derivatization Reagent for HPLC-MS Analysis of Biological Organic Acids.

Small molecules containing carboxylic acid functional groups are ubiquitous throughout biology, playing vital roles in biological chemistry ranging from energy metabolism to cellular signaling. This paper describes a new derivatization reagent, 4-bromo-N-methylbenzylamine, which was selected for its potential to derivatize mono-, di- and tri-carboxylic acids, such as the intermediates of the tricarboxylic acid (TCA) cycle. This derivatization procedure facilitated the use of positive electrospray ionization (ESI) tandem mass spectrometry (MS/MS) detection of derivatized species allowing for clear identification thanks to the easily recognizable isotope pattern of the incorporated bromine. A liquid chromatography (LC)-MS/MS method was developed which provided limits of detection between 0.2 and 44 µg L-1 in under 6 min, depending on the analyte and total analysis time. This method was successfully applied in both in vitro and in vivo models.

DNA methylation-based classification and grading system for meningioma: a multicentre, retrospective analysis.

BACKGROUND: The WHO classification of brain tumours describes 15 subtypes of meningioma. Nine of these subtypes are allotted to WHO grade I, and three each to grade II and grade III. Grading is based solely on histology, with an absence of molecular markers. Although the existing classification and grading approach is of prognostic value, it harbours shortcomings such as ill-defined parameters for subtypes and grading criteria prone to arbitrary judgment. In this study, we aimed for a comprehensive characterisation of the entire molecular genetic landscape of meningioma to identify biologically and clinically relevant subgroups. METHODS: In this multicentre, retrospective analysis, we investigated genome-wide DNA methylation patterns of meningiomas from ten European academic neuro-oncology centres to identify distinct methylation classes of meningiomas. The methylation classes were further characterised by DNA copy number analysis, mutational profiling, and RNA sequencing. Methylation classes were analysed for progression-free survival outcomes by the Kaplan-Meier method. The DNA methylation-based and WHO classification schema were compared using the Brier prediction score, analysed in an independent cohort with WHO grading, progression-free survival, and disease-specific survival data available, collected at the Medical University Vienna (Vienna, Austria), assessing methylation patterns with an alternative methylation chip. FINDINGS: We retrospectively collected 497 meningiomas along with 309 samples of other extra-axial skull tumours that might histologically mimic meningioma variants. Unsupervised clustering of DNA methylation data clearly segregated all meningiomas from other skull tumours. We generated genome-wide DNA methylation profiles from all 497 meningioma samples. DNA methylation profiling distinguished six distinct clinically relevant methylation classes associated with typical mutational, cytogenetic, and gene expression patterns. Compared with WHO grading, classification by individual and combined methylation classes more accurately identifies patients at high risk of disease progression in tumours with WHO grade I histology, and patients at lower risk of recurrence among WHO grade II tumours (p=0·0096) from the Brier prediction test). We validated this finding in our independent cohort of 140 patients with meningioma. INTERPRETATION: DNA methylation-based meningioma classification captures clinically more homogenous groups and has a higher power for predicting tumour recurrence and prognosis than the WHO classification. The approach presented here is potentially very useful for stratifying meningioma patients to observation-only or adjuvant treatment groups. We consider methylation-based tumour classification highly relevant for the future diagnosis and treatment of meningioma. FUNDING: German Cancer Aid, Else Kröner-Fresenius Foundation, and DKFZ/Heidelberg Institute of Personalized Oncology/Precision Oncology Program.