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BackgroundThere is a need for medical education on health care transitions for youth with special health care needs. The Texas Transition Toolkit (the tool) supports providers through a one-stop shop for researching literature on care transitions, a catalog of care transition tools, and guides for developing care transition programs. ObjectiveThis study aims to assess the functionality and usability of the tool with providers working with transition-aged children and youth with special health care needs (representative users). MethodsThe tool was evaluated using a triangulated mixed methods case study approach consisting of a concurrent think-aloud phase, a satisfaction survey, and a survey of problem relevance and task performance to operationalize and capture functionality and usability. Our mixed methods deep dive into the functionality and usability of the tool focused on 10 representative users from one medical home in Texas and 5 website design experts. ResultsRepresentative users found the tool to be highly relevant, as demonstrated by the satisfaction score for relevance (138/150, 92%). According to the users, the tool provided comprehensive information related to health care transitions for youth with special health care needs, with a satisfaction score of 87.3% (131/150) for comprehensive. Overall satisfaction with the tool was high at 81.92% (1065/1300) with a cutoff score of 73.33% (953.4/1300) indicating high satisfaction, but users reported relatively lower satisfaction with search (114/150, 76%) and navigation (ease of use: 114/150, 76%; hyperlinks: 163/200, 81.5%; structure: 159/200, 79.5%). They experienced search- and navigation-related problems (total problems detected: 21/31, 68%) and, based on quality checks, had a relatively low task completion rate for tasks involving finding information (60/80, 75%), which required searching and navigation. The problems identified around search and navigation functionality were relevant (relevance scores ranging from 14.5 to 22, with a cutoff score of 11.7 indicating relevance). ConclusionsThe tool may help bridge the gaps in training on health care transitions for youth with special health care needs in US medical education. The tool can be used to create structured protocols to help improve provider knowledge, collaboration across pediatric and adult care providers, and the continuity of care as youth with special health care needs transition from pediatric to adult care. The results provided a road map for optimizing the tool and highlighted the importance of evaluating eHealth technologies with representative users.

The hepatitis C virus (HCV) infects more than 180 million people worldwide, with long-term consequences including liver failure and hepatocellular carcinoma. Quercetin bioflavonoids can decrease HCV production in tissue culture, in part through inhibition of heat shock proteins. If quercetin demonstrates safety and antiviral activity in patients, then it could be developed into an inexpensive HCV treatment for third world countries or other affected populations that lack financial means to cover the cost of mainstream antivirals. A phase 1 dose escalation study was performed to evaluate the safety of quercetin in 30 untreated patients with chronic HCV infection and to preliminarily characterize quercetin's potential in suppressing viral load and/or liver injury. Quercetin displayed safety in all trial participants. Additionally, 8 patients showed a "clinically meaningful" 0.41-log viral load decrease. There was a positive correlation (r = 0.41, p = 0.03) indicating a tendency for HCV decrease in patients with a lower ratio of plasma quercetin relative to dose. No significant changes in aspartate transaminase and alanine transaminase were detected. In conclusion, quercetin exhibited safety (up to 5 g daily) and there was a potential for antiviral activity in some hepatitis C patients.

Tourette disorder (TD) is poorly understood, despite affecting 1/160 children. A lack of animal models possessing construct, face, and predictive validity hinders progress in the field. We used CRISPR/Cas9 genome editing to generate mice with mutations orthologous to human de novo variants in two high-confidence Tourette genes, CELSR3 and WWC1. Mice with human mutations in Celsr3 and Wwc1 exhibit cognitive and/or sensorimotor behavioral phenotypes consistent with TD. Sensorimotor gating deficits, as measured by acoustic prepulse inhibition, occur in both male and female Celsr3 TD models. Wwc1 mice show reduced prepulse inhibition only in females. Repetitive motor behaviors, common to Celsr3 mice and more pronounced in females, include vertical rearing and grooming. Sensorimotor gating deficits and rearing are attenuated by aripiprazole, a partial agonist at dopamine type II receptors. Unsupervised machine learning reveals numerous changes to spontaneous motor behavior and less predictable patterns of movement. Continuous fixed-ratio reinforcement shows that Celsr3 TD mice have enhanced motor responding and reward learning. Electrically evoked striatal dopamine release, tested in one model, is greater. Brain development is otherwise grossly normal without signs of striatal interneuron loss. Altogether, mice expressing human mutations in high-confidence TD genes exhibit face and predictive validity. Reduced prepulse inhibition and repetitive motor behaviors are core behavioral phenotypes and are responsive to aripiprazole. Enhanced reward learning and motor responding occur alongside greater evoked dopamine release. Phenotypes can also vary by sex and show stronger affection in females, an unexpected finding considering males are more frequently affected in TD. [ABSTRACT FROM AUTHOR]

Food insecurity is a risk factor for morbidity and mortality leading to high medical expenditures, but race/ethnicity was used as adjustments in the literature. The study sought to use race/ethnicity as a key predictor to compare racial differences in associations between food insecurity and expenditures of seven health services among non-institutionalized adults. This cross-sectional study used Medical Expenditure Panel Survey that collects information on food insecurity in 2016 (n=24,179) and 2017 (n=22,539). We examined the association between race/ethnicity and food insecurity status and documented the extent to which impacts of food insecurity on medical expenditures varied by race/ethnicity. We fit multivariable models for each racial group, adjusting for states, age, gender, insurance, and education. Adults older than 18 years were included. The results show that blacks experienced an inter-racial disparity in food insecurity whereas Hispanics experienced intra-racial disparity. A higher percentage of blacks (28.7%) reported at least one type of food insecurity (11.2% of whites). Around 20% of blacks reported being worried about running out of food and the corresponding number is 8.4% among whites. Hispanics reported more food insecurity issues than whites. Moreover, food insecurity is positively associated with expenditures on emergency room utilization (99% increase for other races vs. 51% increase for whites) but is negatively associated with dental care utilization (43% decrease for blacks and 44% for whites). Except for Hispanics, prescription expenditure has the most positive association with food insecurity, and food insecure blacks are the only group that did not significantly use home health. The study expanded our understanding of food insecurity by investigating how it affected seven types of medical expenditures for each of four racial populations. An interdisciplinary effort is needed to enhance the food supply for minorities. Policy interventions to address intra-racial disparities among Hispanics and inter-racial disparities among African Americans are imperative to close the gap. [ABSTRACT FROM AUTHOR]

Immunoglobulin heavy chain variable region exons are assembled in progenitor-B cells, from VH, D, and JH gene segments located in separate clusters across the Igh locus. RAG endonuclease initiates V(D)J recombination from a JH-based recombination center (RC). Cohesin-mediated extrusion of upstream chromatin past RC-bound RAG presents Ds for joining to JHs to form a DJH-RC. Igh has a provocative number and organization of CTCF-binding elements (CBEs) that can impede loop extrusion. Thus, Igh has two divergently oriented CBEs (CBE1 and CBE2) in the IGCR1 element between the VH and D/JH domains, over 100 CBEs across the VH domain convergent to CBE1, and 10 clustered 3'Igh-CBEs convergent to CBE2 and VH CBEs. IGCR1 CBEs segregate D/JH and VH domains by impeding loop extrusion-mediated RAG-scanning. Downregulation of WAPL, a cohesin unloader, in progenitor-B cells neutralizes CBEs, allowing DJH-RC-bound RAG to scan the VH domain and perform VH-to-DJH rearrangements. To elucidate potential roles of IGCR1-based CBEs and 3'Igh-CBEs in regulating RAG-scanning and elucidate the mechanism of the ordered transition from D-to-JH to VH-to-DJH recombination, we tested effects of inverting and/or deleting IGCR1 or 3'Igh-CBEs in mice and/or progenitor-B cell lines. These studies revealed that normal IGCR1 CBE orientation augments RAG-scanning impediment activity and suggest that 3'Igh-CBEs reinforce ability of the RC to function as a dynamic loop extrusion impediment to promote optimal RAG scanning activity. Finally, our findings indicate that ordered V(D)J recombination can be explained by a gradual WAPL downregulation mechanism in progenitor-B cells as opposed to a strict developmental switch. [ABSTRACT FROM AUTHOR]

Classical nonhomologous end joining (C-NHEJ) repairs DNA doublestrand breaks (DSBs) throughout interphase but predominates in G1 phase when homologous recombination is unavailable. Complexes containing the Ku70/80 ("Ku") and XRCC4/ligase IV (Lig4) core C-NHEJ factors are required, respectively, for sensing and joining DSBs. While XRCC4/Lig4 are absolutely required for joining RAG1/2 endonuclease ("RAG")-initiated DSBs during V(D)J recombination in G1-phase progenitor lymphocytes, cycling cells deficient for XRCC4/Lig4 also can join chromosomal DSBs by alternative endjoining (A-EJ) pathways. Restriction of V(D)J recombination by XRCC4/Lig4-mediated joining has been attributed to RAG shepherding V(D)J DSBs exclusively into the C-NHEJ pathway. Here, we report that A-EJ of DSB ends generated by RAG1/2, Cas9:gRNA, and Zinc finger endonucleases in Lig4-deficient G1-arrested progenitor B cell lines is suppressed by Ku. Thus, while diverse DSBs remain largely as free broken ends in Lig4-deficient G1-arrested progenitor B cells, deletion of Ku70 increases DSB rejoining and translocation levels to those observed in Ku70-deficient counterparts. Correspondingly, while RAG-initiated V(D)J DSB joining is abrogated in Lig4-deficient G1-arrested progenitor B cell lines, joining of RAGgenerated DSBs in Ku70-deficient and Ku70/Lig4 double-deficient lines occurs through a translocation-like A-EJ mechanism. Thus, in G1-arrested, Lig4-deficient progenitor B cells are functionally endjoining suppressed due to Ku-dependent blockage of A-EJ, potentially in association with G1-phase down-regulation of Lig1. Finally, we suggest that differential impacts of Ku deficiency versus Lig4 deficiency on V(D)J recombination, neuronal apoptosis, and embryonic development results from Ku-mediated inhibition of A-EJ in the G1 cell cycle phase in Lig4-deficient developing lymphocyte and neuronal cells. [ABSTRACT FROM AUTHOR]

Background: The failure of most clinical Alzheimer's disease (AD) trials has been partially attributed to the lack of translatability of current AD mouse models to human patients. A recently developed model of familial AD (fAD), expressing Swedish, Arctic and Austrian mutations in App (hAbetaSAA), has been shown to be a useful amyloidogenic model which recapitulates many aspects of human AD, including plaque distribution and microglial transcriptional changes. Our aim is to further characterize the hAbetaSAA model and compare it to the widely used 5xFAD transgenic model. Method: Motion Sequencing (MoSeq) software was used to model the underlying structure of spontaneous behaviors recorded in an open field in hAbetaSAA and 5xFAD mice longitudinally from 10 to 18 months of age. Aged hAbetaSAA and 5xFAD brain tissue was used for spatial transcriptomic/proteomic profiling, performed by Nanostring's GeoMx®, which allowed for quantification of gene and protein expression in plaque‐associated and non‐plaque‐associated regions of interest. A fluorophore‐conjugated amyloid antibody (Methoxy‐X04) was administered to cohorts of hAbetaSAA and 5xFAD prior to harvest at various ages. An additional fluorophore‐conjugated antibody (lectin Dylight®594) was administered to 19‐month‐old cohorts permitting visualization of cerebral amyloid angiopathy (CAA). Whole brains were sectioned/imaged using Serial Two‐Photon Tomography on the TissueCyte (TissueVision), creating indexed tissue sections and high‐resolution 3D models of each brain. Subsequent rounds of staining permitted characterization of disease‐associated microglia (DAM) and dystrophic neurites. An independent cohort was evaluated for EEG telemetry. Result: MoSeq revealed divergent behaviors of 5xFAD and hAbetaSAA mice suggesting differences in behavioral phenotypes of these two models. Preliminary GeoMx data shows upregulation of DAM genes localized to plaques in hAbetaSAA homozygotes as identified by RNA‐seq; correlating 5xFAD data is in progress. Further assessments of bulk RNA‐Seq, amyloid distribution, CAA burden and cortical EEG spectral analysis are underway. Conclusion: Comparison of hAbetaSAA and 5xFAD using innovative modes of assessment showcase hAbetaSAA as an amyloidogenic mouse model of fAD that aligns more closely with human than 5xFAD. This model is available for preclinical research with no licensing restrictions and is devoid of artifacts related to transgenic overexpression, positioning it as an improved mouse model for studying fAD mutations. [ABSTRACT FROM AUTHOR]

An effective adaptive immune system depends on the ability of developing B and T cells to generate diverse immunoglobulin (Ig) and T cell receptor repertoires, respectively. Such diversity is achieved through a programmed somatic recombination process whereby germline V, D, and J segments of antigen receptor loci are assembled to form the variable region V(D)J exons of Ig and TCRs. Studies of this process, termed V(D)J recombination, have provided key insights into our understanding of a variety of general gene regulatory and DNA repair processes over the last several decades. V(D)J recombination is initiated by the RAG endonuclease which generates DNA double-stranded breaks at the borders of V, D, and J segments. In this review, we cover recent work that has elucidated RAG structure and work that revealed that RAG has a novel chromatin scanning activity, likely mediated by chromatin loop extrusion, that contributes to its ability to locate V, D, J gene segment substrates within large chromosomal loop domains bounded by CTCF-binding elements (CBEs). This latter function, coupled with the role CBE-based chromatin loop domains and subdomains within them play in focusing V(D)J recombination activity within antigen receptor loci, provide mechanistic explanations for long-standing questions regarding V(D)J segment usage diversification and in limiting potentially deleterious off-target RAG-initiated recombination events genome-wide. This review will focus mainly on studies of the mouse Ig heavy chain locus, but the principles described also apply to other Ig loci and to TCR loci in mice and humans. [ABSTRACT FROM AUTHOR]

Gene targeting in human somatic cells is of importance because it can be used to either delineate the loss-of-function phenotype of a gene or correct a mutated gene back to wild-type. Both of these outcomes require a form of DNA double-strand break (DSB) repair known as homologous recombination (HR). The mechanism of HR leading to gene targeting, however, is not well understood in human cells. Here, we demonstrate that a two-end, ends-out HR intermediate is valid for human gene targeting. Furthermore, the resolution step of this intermediate occurs via the classic DSB repair model of HR while synthesis-dependent strand annealing and Holliday Junction dissolution are, at best, minor pathways. Moreover, and in contrast to other systems, the positions of Holliday Junction resolution are evenly distributed along the homology arms of the targeting vector. Most unexpectedly, we demonstrate that when a meganuclease is used to introduce a chromosomal DSB to augment gene targeting, the mechanism of gene targeting is inverted to an ends-in process. Finally, we demonstrate that the anti-recombination activity of mismatch repair is a significant impediment to gene targeting. These observations significantly advance our understanding of HR and gene targeting in human cells. [ABSTRACT FROM AUTHOR]

Summary: Methylation of histone H3 lysine 4 by the Set1 subunit of COMPASS correlates with active transcription. Here, we show that Set1 levels are regulated by protein degradation in response to multiple signals. Set1 levels are greatly reduced when COMPASS recruitment to genes, H3K4 methylation, or transcription is blocked. The degradation sequences map to N-terminal regions that overlap a previously identified autoinhibitory domain, as well as the catalytic domain. Truncation mutants of Set1 that cause under- or overexpression produce abnormal H3K4 methylation patterns on transcribed genes. Surprisingly, SAGA-dependent genes are more strongly affected than TFIID-dependent genes, reflecting differences in their chromatin dynamics. We propose that careful tuning of Set1 levels by regulated degradation is critical for the establishment and maintenance of proper H3K4 methylation patterns. [Copyright &y& Elsevier]

This study examined the effects of age, target location, and stimulus speed on coincidence-anticipation timing in a catching task. Males aged 11 to 18 years made simulated catching movements toward a light stimulus that rapidly approached the head or chest at various speeds. Coincidence-anticipation timing accuracy, movement onset times, and movement times did not differ by age. However, 17- to 18-year-olds exhibited significantly faster movement speeds than 14- to 16-year-olds. Target location (head or chest) did not affect coincidence-anticipation timing accuracy or movement speed. However, movements toward the head were initiated earlier and took longer than movements to the chest. Finally, stimulus speed had statistically significant effects on all measures: faster stimuli were associated with longer delays in coincidence-anticipation timing responses, earlier movement onset times, shorter movement times, and faster movement speeds. These results underscore the adaptability of coincidence-anticipation timing abilities for responding to stimuli under varying temporal and spatial constraints. [ABSTRACT FROM AUTHOR]

Examines a modified two-sector model to determine the effects of financial liberalization on allocative efficiency among financial markets in Taiwan. Identification of divisions in a financial sector; Emphasis on the linear relationship between deposit rate and loan rate of a curb market; Implications of the existence of a curb financial market for allocative efficiency.

Behavior is shaped by both the internal state of an animal and its individual behavioral biases. Rhythmic variation in gonadal hormones during the estrous cycle is a defining feature of the female internal state, one that regulates many aspects of sociosexual behavior. However, it remains unclear whether estrous state influences spontaneous behavior and, if so, how these effects might relate to individual behavioral variation. Here, we address this question by longitudinally characterizing the open-field behavior of female mice across different phases of the estrous cycle, using unsupervised machine learning to decompose spontaneous behavior into its constituent elements. 1,2,3,4 We find that each female mouse exhibits a characteristic pattern of exploration that uniquely identifies it as an individual across many experimental sessions; by contrast, estrous state only negligibly impacts behavior, despite its known effects on neural circuits that regulate action selection and movement. Like female mice, male mice exhibit individual-specific patterns of behavior in the open field; however, the exploratory behavior of males is significantly more variable than that expressed by females both within and across individuals. These findings suggest underlying functional stability to the circuits that support exploration in female mice, reveal a surprising degree of specificity in individual behavior, and provide empirical support for the inclusion of both sexes in experiments querying spontaneous behaviors. • Spontaneous behavior of female mice is only negligibly affected by estrous state • Females and males exhibit strongly individualized patterns of exploration • Female spontaneous behavior is less variable than male behavior Levy et al. track open-field behavior of female mice over weeks and find that behavior reflects individual identity far more than estrous state. Open-field exploration patterns in males are much more variable than in females, arguing for the inclusion of both sexes in studies of spontaneous behaviors. [ABSTRACT FROM AUTHOR]

Mammalian SWI/SNF (BAF) chromatin remodelers play dosage-sensitive roles in many human malignancies and neurologic disorders. The gene encoding the BAF subunit actin-like 6a (ACTL6A) is amplified early in the development of many squamous cell carcinomas (SCCs), but its oncogenic role remains unclear. Here we demonstrate that ACTL6A overexpression leads to its stoichiometric assembly into BAF complexes and drives their interaction and engagement with specific regulatory regions in the genome. In normal epithelial cells, ACTL6A was substoichiometric to other BAF subunits. However, increased ACTL6A levels by ectopic expression or in SCC cells led to near saturation of ACTL6A within BAF complexes. Increased ACTL6A occupancy enhanced polycomb opposition genome-wide to activate SCC genes and facilitated the co-dependent loading of BAF and TEAD-YAP complexes on chromatin. Both mechanisms appeared to be critical and function as a molecular AND gate for SCC initiation and maintenance, thereby explaining the specificity of the role of ACTL6A amplification in SCCs. [Display omitted] • ACTL6A occupancy in the BAF complex is substoichiometric in normal epithelial cells • SCC cells upregulate ACTL6A, thus increasing ACTL6A assembly with the BAF complex • ACTL6A mediates co-dependent chromatin loading of BAF and TEAD-YAP complexes • ACTL6A upregulation counteracts polycomb-mediated repression at SCC signature genes Chang et al. find that ACTL6A plays a dosage-sensitive role underlying squamous cell carcinoma (SCC). Early in the course of the development of SCC, ACTL6A gene amplification increases its normally unsaturated occupancy within BAF complexes, leading to epigenetic derepression by polycomb redistribution and increased chromatin loading of TEAD-YAP. [ABSTRACT FROM AUTHOR]

This study examined the effects of age, target location, and stimulus speed on coincidence-anticipation timing in a catching task. Males aged 11 to 18 years made simulated catching movements toward a light stimulus that rapidly approached the head or chest at various speeds. Coincidence-anticipation timing accuracy, movement onset times, and movement times did not differ by age. However, 17- to 18-year-olds exhibited significantly faster movement speeds than 14- to 16-year-olds. Target location (head or chest) did not affect coincidence-anticipation timing accuracy or movement speed. However, movements toward the head were initiated earlier and took longer than movements to the chest. Finally, stimulus speed had statistically significant effects on all measures: faster stimuli were associated with longer delays in coincidence-anticipation timing responses, earlier movement onset times, shorter movement times, and faster movement speeds. These results underscore the adaptability of coincidence-anticipation timing abilities for responding to stimuli under varying temporal and spatial constraints. [ABSTRACT FROM AUTHOR]

Summary The design of immunogens that elicit broadly reactive neutralizing antibodies (bnAbs) has been a major obstacle to HIV-1 vaccine development. One approach to assess potential immunogens is to use mice expressing precursors of human bnAbs as vaccination models. The bnAbs of the VRC01-class derive from the IGHV1-2 immunoglobulin heavy chain and neutralize a wide spectrum of HIV-1 strains via targeting the CD4 binding site of the envelope glycoprotein gp120. We now describe a mouse vaccination model that allows a germline human IGHV1-2 ∗ 02 segment to undergo normal V(D)J recombination and, thereby, leads to the generation of peripheral B cells that express a highly diverse repertoire of VRC01-related receptors. When sequentially immunized with modified gp120 glycoproteins designed to engage VRC01 germline and intermediate antibodies, IGHV1-2 ∗ 02-rearranging mice, which also express a VRC01-antibody precursor light chain, can support the affinity maturation of VRC01 precursor antibodies into HIV-neutralizing antibody lineages. [ABSTRACT FROM AUTHOR]

Spontaneous mouse behavior is composed from repeatedly used modules of movement (e.g., rearing, running or grooming) that are flexibly placed into sequences whose content evolves over time. By identifying behavioral modules and the order in which they are expressed, researchers can gain insight into the effect of drugs, genes, context, sensory stimuli and neural activity on natural behavior. Here we present a protocol for performing Motion Sequencing (MoSeq), an ethologically inspired method that uses three-dimensional machine vision and unsupervised machine learning to decompose spontaneous mouse behavior into a series of elemental modules called 'syllables'. This protocol is based upon a MoSeq pipeline that includes modules for depth video acquisition, data preprocessing and modeling, as well as a standardized set of visualization tools. Users are provided with instructions and code for building a MoSeq imaging rig and acquiring three-dimensional video of spontaneous mouse behavior for submission to the modeling framework; the outputs of this protocol include syllable labels for each frame of the video data as well as summary plots describing how often each syllable was used and how syllables transitioned from one to the other. In addition, we provide instructions for analyzing and visualizing the outputs of keypoint-MoSeq, a recently developed variant of MoSeq that can identify behavioral motifs from keypoints identified from standard (rather than depth) video. This protocol and the accompanying pipeline significantly lower the bar for users without extensive computational ethology experience to adopt this unsupervised, data-driven approach to characterize mouse behavior., (© 2024. Springer Nature Limited.)

Tourette disorder (TD) is poorly understood, despite affecting 1/160 children. A lack of animal models possessing construct, face, and predictive validity hinders progress in the field. We used CRISPR/Cas9 genome editing to generate mice with mutations orthologous to human de novo variants in two high-confidence Tourette genes, CELSR3 and WWC1 . Mice with human mutations in Celsr3 and Wwc1 exhibit cognitive and/or sensorimotor behavioral phenotypes consistent with TD. Sensorimotor gating deficits, as measured by acoustic prepulse inhibition, occur in both male and female Celsr3 TD models. Wwc1 mice show reduced prepulse inhibition only in females. Repetitive motor behaviors, common to Celsr3 mice and more pronounced in females, include vertical rearing and grooming. Sensorimotor gating deficits and rearing are attenuated by aripiprazole, a partial agonist at dopamine type II receptors. Unsupervised machine learning reveals numerous changes to spontaneous motor behavior and less predictable patterns of movement. Continuous fixed-ratio reinforcement shows that Celsr3 TD mice have enhanced motor responding and reward learning. Electrically evoked striatal dopamine release, tested in one model, is greater. Brain development is otherwise grossly normal without signs of striatal interneuron loss. Altogether, mice expressing human mutations in high-confidence TD genes exhibit face and predictive validity. Reduced prepulse inhibition and repetitive motor behaviors are core behavioral phenotypes and are responsive to aripiprazole. Enhanced reward learning and motor responding occur alongside greater evoked dopamine release. Phenotypes can also vary by sex and show stronger affection in females, an unexpected finding considering males are more frequently affected in TD., Competing Interests: Competing interests statement:The authors declare no competing interest.

Immunoglobulin heavy chain variable region exons are assembled in progenitor-B cells, from V H , D, and J H gene segments located in separate clusters across the Igh locus. RAG endonuclease initiates V(D)J recombination from a J H -based recombination center (RC). Cohesin-mediated extrusion of upstream chromatin past RC-bound RAG presents Ds for joining to J H s to form a DJ H -RC. Igh has a provocative number and organization of CTCF-binding elements (CBEs) that can impede loop extrusion. Thus, Igh has two divergently oriented CBEs (CBE1 and CBE2) in the IGCR1 element between the V H and D/J H domains, over 100 CBEs across the V H domain convergent to CBE1, and 10 clustered 3' Igh -CBEs convergent to CBE2 and V H CBEs. IGCR1 CBEs segregate D/J H and V H domains by impeding loop extrusion-mediated RAG-scanning. Downregulation of WAPL, a cohesin unloader, in progenitor-B cells neutralizes CBEs, allowing DJ H -RC-bound RAG to scan the V H domain and perform V H -to-DJ H rearrangements. To elucidate potential roles of IGCR1-based CBEs and 3' Igh -CBEs in regulating RAG-scanning and elucidate the mechanism of the ordered transition from D-to-J H to V H -to-DJ H recombination, we tested effects of inverting and/or deleting IGCR1 or 3' Igh -CBEs in mice and/or progenitor-B cell lines. These studies revealed that normal IGCR1 CBE orientation augments RAG-scanning impediment activity and suggest that 3' Igh -CBEs reinforce ability of the RC to function as a dynamic loop extrusion impediment to promote optimal RAG scanning activity. Finally, our findings indicate that ordered V(D)J recombination can be explained by a gradual WAPL downregulation mechanism in progenitor-B cells as opposed to a strict developmental switch.

Cell segmentation is a common step in cell behavior analysis. Reliably and automatically segmenting cells in microscopy images remains challenging, especially in differential inference contrast microscopy images and phase-contrast microscopy images. In this paper, we propose a deep learning solution combining a Mask RCNN architecture with Shape-Aware Loss to produce cell instance segmentation. Our approach outperforms prior works in cell segmentation, achieving an IOU of 91.91% on the DIC-C2DH-HeLa dataset and an IOU of 94.93 % on the PhC-C2DH-U373 dataset. Our framework can calculate cell instance segmentation masks from both types of microscopy images without any additional post-processing.Clinical Relevance - The proposed approach produces accurate instance segmentation in Differential Inference Contrast and Phase-Contrast microscopy images. The segmentation results can be reliably used in cell behavior analysis and cell tracking.

Purpose: The purpose of this study is to describe a novel computerized diabetes education tool and explore factors influencing self-selection and use among primarily Hispanic patients diagnosed with type 2 diabetes in south Texas., Methods: Study participants included 953 adult patients with type 2 diabetes enrolled in a diabetes education program between July 1, 2016, and June 30, 2017. Participants were asked to choose either a new technology-based diabetes education tool with a touch-screen device or a traditional face-to-face education method. Multivariate logistic regression analysis was applied to identify factors associated with adopting the computerized diabetes education tool among the patients., Results: When comparing technology-based tool adopters and nonadopters, several demographic and health-related factors differentiated technology use in bivariate analyses. The multivariate logistic regression model showed that Hispanic patients were less likely to choose a technology-based tool. Patients who perceived their health status as excellent/good were more likely to adopt the technologic education method than those with fair/poor perceived health status. A1C level was negatively associated with self-selection of technology., Conclusions: Specific demographic and health-related characteristics are significant contributing factors to patients' adoption of a technology-based diabetes education tool. Health care providers can utilize these findings to target and refer specific patients to a computerized diabetes education tool for more effective diabetes care and to optimize technology adoption success.

Purpose: The opioid crisis is devastating rural America, but findings of opioid utilization vary among previous studies. Previous studies were focused on misuse behaviors or overdose issues. This study will focus on the number of pills and prescriptions that rural and urban adults received., Methods: Using the adult data of the 2011-2016 Medical Expenditure Panel Survey, we compared rural-urban differences in likelihood of using opioids and actual utilization. Multivariate models were further adjusted for predisposing, enabling and need factors., Results: During 2011-2016, opioid utilization decreased in both urban and rural areas. However, rural adults were still more likely to have a prescription, and among users, rural adult prescription pill count was higher than urban counterparts. The rural-urban difference was not significant after adjusting for covariates, indicating that personal and contextual characteristics account for more variations in utilization than rurality., Conclusions: Strategies to improve pain management without causing opioid addiction and overdose deaths are imperative. The findings of unadjusted analyses suggest: (1) providing counseling to teach rural adults to store opioids in a locked container, not share medication with others and safely dispose of unused pills; (2) reinforcing the mail-back program or giving patients a specially-designed package to neutralize the drugs; and (3) if a community-based drug-disposal program is not available, educating to remove labeling from the bottle, mix the drugs with an unpleasant substance, and place the drugs and unpleasant substance in the garbage separate from the bottle. The findings of adjusted analyses indicate that another study will be helpful to explore the associations between personal characteristics and opioid utilization in depth.

Objective: Despite the high prevalence of chronic multisite pain, there is little consensus on methods to characterize it. Commonly used assessments report only one dimension of pain, that is, intensity, thus ignoring the spatial aspect of pain. We developed a novel pain quantification index, the Integrated Pain Quantification Index (IPQI), on a scale of 0 to 1 that integrates multiple distinct pain measures into a single value, thus representing multidimensional pain information with a single value., Design: Single-visit, noninterventional, epidemiological study., Setting: Fourteen outpatient multidisciplinary pain management programs., Patients: Patients with chronic pain of the trunk and/or limbs for at least six months with average overall pain intensity of at least 5 on the numeric rating scale., Methods: Development of IPQI was performed in a large population (N = 810) of chronic pain patients from the Multiple Areas of Pain (MAP) study., Results: Prevalence of two or more noncontiguous painful areas was at 88.3% (95% confidence interval [CI] = 0.86-0.90), with a mean of 6.3 areas (SD = 5.57 areas). Prevalence of more than 10% body area in pain was at 52.8% (95% CI = 0.49-0.56), with a mean at 16.1% (17.16%). On average, IPQI values were near the middle of the scale, with mean and median IPQI at 0.52 (SD = 0.13) and 0.55, respectively. The IPQI was generalizable and clinically relevant across all domains recommended by the Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials., Conclusions: IPQI provided a single pain score for representing complex, multidimensional pain information on one scale and has implications for comparing pain populations across longitudinal clinical trials., (© 2018 American Academy of Pain Medicine. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

An effective adaptive immune system depends on the ability of developing B and T cells to generate diverse immunoglobulin (Ig) and T cell receptor repertoires, respectively. Such diversity is achieved through a programmed somatic recombination process whereby germline V, D, and J segments of antigen receptor loci are assembled to form the variable region V(D)J exons of Ig and TCRs. Studies of this process, termed V(D)J recombination, have provided key insights into our understanding of a variety of general gene regulatory and DNA repair processes over the last several decades. V(D)J recombination is initiated by the RAG endonuclease which generates DNA double-stranded breaks at the borders of V, D, and J segments. In this review, we cover recent work that has elucidated RAG structure and work that revealed that RAG has a novel chromatin scanning activity, likely mediated by chromatin loop extrusion, that contributes to its ability to locate V, D, J gene segment substrates within large chromosomal loop domains bounded by CTCF-binding elements (CBEs). This latter function, coupled with the role CBE-based chromatin loop domains and subdomains within them play in focusing V(D)J recombination activity within antigen receptor loci, provide mechanistic explanations for long-standing questions regarding V(D)J segment usage diversification and in limiting potentially deleterious off-target RAG-initiated recombination events genome-wide. This review will focus mainly on studies of the mouse Ig heavy chain locus, but the principles described also apply to other Ig loci and to TCR loci in mice and humans., (© 2018 Elsevier Inc. All rights reserved.)

Taiwan was an endemic area for hepatitis B virus (HBV) infection, and related liver diseases cause a significant drain of public resources. To control the endemic, a nation-wide newborn vaccination program was started in 1985. We reviewed the results of the annual survey for HBV surface antigen (HBsAg) performed in freshmen class of two high schools in Hualien, eastern Taiwan, from 1991 to 2001. A total of 10,194 students, most of them 15 years old, were tested for serum HBsAg using enzyme immunoassays. There is a significant trend (P < 0.0001) of decreasing HBsAg carrier rate from 20.3 to 4.4% in males and 14.3% to 2.4% in females, respectively, over 11 years. The HBsAg carrier rate was 16.0-20.3% in students surveyed during 1991-1993 (born more than 6 years before the start of the national vaccination program), which decreased to 7.7-11.9% during 1994-1999 (born 1-6 years before the program). It further declined to 4.7% and 3.4% in 2000 and 2001 (born after the start of the program). The HBsAg carrier rate in male students was significantly higher than that in female students in most of the years. The HBV newborn vaccination program not only successfully prevented most of the perinatal transmission of HBV but also reduced horizontal transmission of HBV to children born up to 6 years before the start of the program. Also, the protection persisted for at least 15 years., (Copyright 2003 Wiley-Liss, Inc.)