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by Viral D. Oza, Colin S. Williams, Jessica S. Blackburn
The Genetically Encoded Death Indicator (GEDI) is a ratiometric, dual-fluorescence biosensor that enables real-time detection of cell death through calcium influx. Originally developed for use in neurodegeneration models, GEDI can be applied to cancer cells to quantify therapy-induced death at single-cell resolution. This protocol details how to generate GEDI-expressing cancer cell lines, empirically determine stress-induced GEDI thresholds using radiation or chemotherapeutic agents, and perform time-resolved imaging and image analysis to track cell fate. This workflow is optimized for high-throughput drug and radiation screening in heterogeneous populations and is especially useful for identifying chemo- and radio-resistant subclones. Key limitations include the need for empirical GEDI threshold calibration for each treatment condition and careful standardization of imaging parameters. The protocol outputs include GEDI ratio values, single-cell time-of-death annotations, and whole-cell morphological data in parallel, which can be linked to downstream applications such as FACS-based isolation of live or dying subpopulations, transcriptomic profiling of resistant clones, or in vivo validation using xenografts or organotypic slice culture.by David A. Green, Jesse M. Maestas, Jessica N. Sanchez, Nathan C. Nieto, Andrew S. Bridges, David K. Garcelon
The San Clemente Island fox (Urocyon littoralis clementae) is classified as a focal species for conservation management by the US Navy. They are considered vulnerable to a variety of vector-borne diseases due to their relatively high population density and low genetic diversity. During the dry (July–November) and wet (December–February) seasons of 2017–2018 we live-trapped 95 foxes and collected ectoparasites to test for the presence of pathogens. We found a significant difference in ectoparasite abundance on foxes between seasons, but no differences associated with sex or age. We found that foxes carried two species of flea (Echidnophaga gallinacea and Orchopeas howardi) and two tick species (Ixodes pacificus and Ixodes jellisoni). No evidence of Borrelia burgdorferi, Anaplasma phagocytophilum, or Borrelia miyamotoi bacteria were found. This paper is the first account of ectoparasite species identification, quantification, and pathogen testing for the San Clemente Island fox subspecies.by Elizabeth Baguley, Madelyn Knaub, Jessica VanDyke, Gideon Hirschfield, Mark G. Swain, Gail Wright, Deirdre McCaughey, Abdel Aziz Shaheen
Pandemic restrictions impacted healthcare, particularly during the first year. We evaluated the impact of the pandemic on quality of life and clinical care among patients with primary biliary cholangitis (PBC). This mixed-methods study administered quality of life surveys (Fear of COVID-19 Scale [FCV-19S], EuroQol 5-dimension 3-level [EQ-5D-3L], 29-item Patient-Reported Outcomes Measurement Instrument Survey [PROMIS-29]) and a PBC Care Delivery questionnaire to 348 Canadian PBC patients, followed by two focus groups with patients (n = 14) and stakeholders (n = 3). Quality of life scores were compared among sub-groups (i.e., care delays and pandemic appointment type) and with various reference populations. Most participants were female (94.0%) and Caucasian (88.2%), with a median age of 63.0 years (IQR: 55.9–71.2). During the pandemic, 75.8% had the majority (≥ 50%) of their hepatologist appointments virtually, but only 22.4% preferred to continue with virtual care post-pandemic. Participants with care delays had worse scores on the FCV-19S (p = 0.014), EQ-5D-3L (p = 0.009), and PROMIS-29 (i.e., fatigue, anxiety, sleep disturbance, ability to participate in social roles and activities, p pby Jessica Liu, Sameer Pandya, Andreas Coppi, H. Patrick Young, Harlan M. Krumholz, Wade L. Schulz, Guannan Gong
BackgroundNear real-time electronic health record (EHR) data offers significant potential for secondary use in research, operations, and clinical care, yet challenges remain in ensuring data quality and stability. While prior studies have assessed retrospective EHR datasets, few have systematically examined the integrity of real-time data for research readiness.
MethodsWe developed an automated benchmarking pipeline to evaluate the stability and completeness of real-time EHR data from the Yale New Haven Health clinical data warehouse, transformed into the OMOP common data model. Twenty-nine weekly snapshots of the EHR collected from July to November 2024 and twenty-two daily snapshots collected from April to May 2025 were analyzed. Benchmarks focused on (1) clinical actions such as patient additions, deletions, and merges; (2) changes in demographic variables (date of birth, gender, race, ethnicity); and (3) stability of discharge information (time and status). A synthetic dataset derived from MIMIC-III was used to validate the benchmarking code prior to large-scale analyses.
ResultsBenchmarking revealed frequent updates due to clinical actions and demographic corrections across consecutive snapshots. Demographic changes were most frequently related to race and ethnicity, highlighting potential workflow and data entry inconsistencies. Discharge time and status values demonstrated instability for several days post-encounter, typically reaching a stable state within 4–7 days. These findings indicate that while near real-time EHR data provide valuable insights, the timing of data stabilization is critical for accurate secondary use.
ConclusionsThis study demonstrates the feasibility of automated benchmarking to assess the integrity of real-time EHR data and identify when such data become analysis ready. Our findings highlight key challenges for secondary use of dynamic clinical data and provide an automated framework that can be applied across health systems to support high-quality research, surveillance, and clinical trial readiness.
by Erica Qureshi, Quynh Doan, Jessica Moe, Steven P. Miller, Garth Meckler, Brett Burstein, Jehannine (J9) Austin
ObjectivesEmergency department (ED) to community (ED2C) programs, which redirect patients from the pediatric ED to community healthcare professionals represent a promising strategy to reduce the impact of non-urgent visits on the pediatric ED. Given an ED2C program’s potential impact on various care professionals, we completed a qualitative study to explore key informants’ attitudes and perceptions of pediatric ED2C programs.
MethodsWe conducted one-on-one semi-structured interviews with key informants in British Columbia, Canada. Participants included: pediatric ED staff – triage nurses and physicians; community professionals – pediatricians and family physicians; and health system leaders responsible for pediatric and emergency care in British Columbia. Interviews were recorded, transcribed verbatim, de-identified, and analyzed using reflexive thematic analysis within an interpretive description framework. A visual model was developed to depict key themes in attitudes and perceptions towards pediatric ED2C programs.
ResultsWe interviewed 24 participants: 6 community professionals, 11 pediatric ED professionals, and 7 healthcare leaders. Participants viewed the ED2C program as a valuable solution to address pediatric system strain provided that systemic barriers are addressed, and both emergency and community settings are equipped with adequate training and resources. Participants emphasized the need for clear guidelines on eligibility and operations to build confidence and enhance program effectiveness.
ConclusionsOur findings suggest there is support for ED2C programs as a means to reduce the impact of non-urgent pediatric ED visits and strengthen community-based care. Successful implementation will require coordinated planning, resource investment, and clear operational frameworks.
by Desirée Valera-Gran, Miriam Hurtado-Pomares, Iris Juárez-Leal, Rocío Muñoz-Sánchez, Irene Campos-Sánchez, Paula Noce, Jessica Piñero, Eva-María Navarrete-Muñoz
BackgroundParents of children with developmental delays (DD) often face significant challenges that affect their mental health and occupational balance. While early intervention services traditionally focus on child development, the occupational needs and well-being of parents remain underexplored. The EQo-Mental project aims to examine the association between parental mental health, occupational balance, and meaningful activity engagement, and to co-develop family-centred strategies that promote well-being in early intervention contexts.
MethodsThis sequential mixed-methods study includes two phases. The quantitative phase will involve approximately 700 parents of children aged 0–6 years attending early intervention centres in Alicante, Spain. This phase comprises two components: (1) the psychometric validation of the Spanish versions of two occupational measures—the Occupational Balance Questionnaire (OBQ-E) and the Engagement in Meaningful Activities Survey (EMAS)—and (2) a cross-sectional analysis examining associations between occupational and mental health outcomes. Participants will complete a sociodemographic questionnaire along with validated self-administered instruments assessing occupational balance, meaningful activity engagement, stress, anxiety, depression, and psychological well-being. In the qualitative phase, participatory sessions and focus groups will be conducted with a subsample of parents and key stakeholders to explore perceived occupational and mental health needs and to co-design actionable strategies for improving occupational balance and family well-being. Participant recruitment began in November 2023 and is ongoing; data collection is expected to be completed by October 2025.
AnalysesPsychometric analyses will first be conducted to evaluate the validity and reliability of the OBQ-E and EMAS. Next, descriptive analyses and multiple regression models adjusted for potential confounders will be used to explore associations between occupational and mental health variables. Phase 2 consists of a participatory-action research process, including discussion groups and a multi-stakeholder focus group. Qualitative data will be analysed using reflexive thematic analysis.
OutcomesFindings from EQo-Mental will inform the design of evidence-based, family-centred strategies that support occupational balance, parental well-being, and engagement in meaningful activities. By addressing the occupational needs of parents, the project seeks to foster more resilient families and strengthen early intervention services through an inclusive, occupation-focused approach.
by Carly E. Hawkins, Thomas P. Hahn, Jessica L. Malisch, Gail L. Patricelli
Males in socially monogamous species can achieve reproductive success through multiple tactics– by defending paternity within the social nest and siring extra-pair offspring, or both. Previous studies have found that sperm morphology may differentially affect fertilization success in extra-pair compared to within-pair matings; therefore, we explored whether sperm morphological traits can predict the probability of success within components of reproductive success. Here, we measured sperm component traits (head length and flagellum length) and derived traits (total length and flagellum:head ratio) in free-living Mountain White-crowned Sparrows (Zonotrichia leucophrys oriantha) and examined how these morphological traits relate to extra-pair and within-pair reproductive components of reproductive success. We found no evidence for correlations between sperm morphology and total seasonal reproductive success. However, we did find that sperm morphology appeared to be associated with whether a male was successful at acquiring extra-pair offspring or defending his own paternity within his nest: males that achieved extra-pair success had longer flagella and longer total length of sperm cells compared to males that did not sire outside of their social nest. In contrast, males that successfully defended all paternity within their social nest tended to have shorter heads and larger flagellum:head ratios compared to males that lost paternity in their social nest. While these patterns suggest that different sperm traits may be linked to success in different components of reproductive success, they should be interpreted with caution given the exploratory nature of this study and limited sample size, and further investigation is warranted.by Sugy Choi, Elizabeth Knopf, Megan A. O’Grady, Ivy Van Domselaar, Jessica Ortiz, Carla King, Charles J. Neighbors, Thomas D’Aunno
BackgroundPregnant and parenting women with substance use disorders (SUDs) face complex and overlapping challenges, including substance use, legal issues, housing instability, and trauma. Effective interorganizational collaboration is critical but often hindered by fragmented care and resource limitations. This study explores the key barriers and facilitators that impact collaborative efforts among healthcare providers, government agencies, and community organizations in addressing SUD among pregnant and parenting women.
MethodsThis qualitative study was conducted in New York State between April 2022 and April 2023. The study focused on organizations that provide services to pregnant and parenting women with SUDs, including government agencies, SUD treatment centers, healthcare settings, and community-based care organizations. Semi-structured, one-on-one interviews were conducted with staff to explore how their organizations coordinate care. Thematic analysis was used to identify patterns related to interorganizational collaboration. Primary data were collected through interviews with 30 staff members across multiple stakeholder groups: child welfare services (n = 8), criminal legal agencies (n = 5), health agencies (n = 3), healthcare service settings (n = 4), SUD treatment programs (n = 6), and community-based organizations (n = 4). Interviews lasted approximately one hour and focused on organizational roles, referral processes, and coordination efforts in serving the target population.
ResultsCollaborative care was primarily facilitated through referral networks, case management teams, and the presence of embedded healthcare professionals. However, these systems were frequently limited by fragmented communication, stigmatizing attitudes, and insufficient resources. Organizational facilitators included co-located healthcare staff within child welfare services and formalized partnerships across sectors. Key barriers included staffing shortages, burnout, and misalignment of organizational goals. At the individual level, collaboration often depended on informal relationships and staff-driven initiatives, though interdisciplinary knowledge gaps remained a significant challenge.
ConclusionsImproving service coordination for pregnant and parenting women with SUDs will require stronger organizational infrastructure, investment in cross-sector communication strategies, and deliberate efforts to address stigma. Future research should explore models that support sustained, formalized interagency partnerships to enhance care integration.
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