bluuebunny/crossref_metadata_2025 huggingface dataset:
The Association of Grit With Burnout Components (Professional Efficacy, Exhaustion, and Cynicism) Among Academic Rheumatologists
Authors: Miyawaki Yoshia , Sada Ken-ei , Shidahara Kenta , Nawachi Shoichi , Asano Yosuke , Katayama Yu , Hayashi Keigo , Katsuyama Eri , Katsuyama Takayuki , Takano-Narazaki Mariko , Matsumoto Yoshinori , Oguro Nao , Yajima Nobuyuki , Ishikawa Yuichi , Sakurai Natsuki , Hidekawa Chiharu , Yoshimi Ryusuke , Ichikawa Takanori , Kishida Dai , Shimojima Yasuhiro , Wada Jun , Kurita Noriaki
Published: May 2023

Objectives

There is a high prevalence of burnout among rheumatologists. Grit, which is defined as possessing perseverance and a passion to achieve long-term goals, is predictive of success in many professions; however, whether grit is associated with burnout remains unclear, especially among academic rheumatologists, who have multiple simultaneous responsibilities. Thus, the purpose of this study was to examine the associations between grit and self-reported burnout components—professional efficacy, exhaustion, and cynicism—in academic rheumatologists.

Methods

This cross-sectional study involved 51 rheumatologists from 5 university hospitals. The exposure was grit, measured using mean scores for the 8-item Short Grit Scale (range, 1–5 [5 = extremely high grit]). The outcome measures were mean scores for 3 burnout domains (exhaustion, professional efficacy, and cynicism; range, 1–6; measured using the 16-item Maslach Burnout Inventory–General Survey). General linear models were fitted with covariates (age, sex, job title [assistant professor or higher vs lower], marital status, and having children).

Results

Overall, 51 physicians (median age, 45 years; interquartile range, 36–57; 76% men) were included. Burnout positivity was found in 68.6% of participants (n = 35/51; 95% confidence interval [CI], 54.1, 80.9). Higher grit was associated with higher professional efficacy (per 1-point increase; 0.51 point; 95% CI, 0.18, 0.84) but not with exhaustion or cynicism. Being male and having children were associated with lower exhaustion (−0.69; 95% CI, −1.28, −0.10; p = 0.02; and −0.85; 95% CI, −1.46, −0.24; p = 0.006). Lower job title (fellow or part-time lecturer) was associated with higher cynicism (0.90; 95% CI, 0.04, 1.75; p = 0.04).

Conclusions

Grit is associated with higher professional efficacy among academic rheumatologists. To prevent burnout among staff, supervisors who manage academic rheumatologists should assess their staff's individual grit.
Characterizing Clinical Trials in Plastic and Reconstructive Surgery
Authors: Cai Lawrence Z. , Patel Ashraf A. , Thirunavu Vineeth M. , Hug Nicholas F. , Song Siyou , Li Jennifer , Barghout Robert R. , Magnani Christopher J. , Turner Brandon E. , Steinberg Jecca R. , Lee Gordon K.
Published: May 2023

Background

Clinical trials form the backbone of evidence-based medicine. ClinicalTrials.gov is the world's largest clinical trial registry, and the state of clinical trials in plastic and reconstructive surgery (PRS) within that database has not been comprehensively studied. To that end, we explored the distribution of therapeutic areas that are under investigation, impact of funding on study design and data reporting, and trends in research patterns of all PRS interventional clinical trials registered with ClinicalTrials.gov.

Methods

Using the ClinicalTrials.gov database, we identified and extracted all clinical trials relevant to PRS that were submitted between 2007 and 2020. Studies were classified based on anatomic locations, therapeutic categories, and specialty topics. Cox proportional hazard was used to calculate adjusted hazard ratios (HRs) for early discontinuation and results reporting.

Results

A total of 3224 trials that included 372,095 participants were identified. The PRS trials grew at an annual rate of 7.9%. The therapeutic classes most represented were wound healing (41.3%) and cosmetics (18.1%). Funding for PRS clinical trials is largely provided through academic institutions (72.7%), while industry and US government constituted a minority. Industry-funded studies were more likely to be discontinued early than those funded by academics (HR, 1.89) or government (HR, 1.92) and to be nonblinded and nonrandomized. Academic-funded studies were the least likely to report results data within 3 years of trial completion (odds ratio, 0.87).

Conclusions

A gulf exists in the representation of different PRS specialties among clinical trials. We highlight the role of funding source in trial design and data reporting to identify a potential source of financial waste and to stress the need for continued appropriate oversight.
Does Early Referral Lead to Early Repair? Quality Improvement in Cleft Care
Authors: Kondra Katelyn , Stanton Eloise , Jimenez Christian , Chen Kevin , Hammoudeh Jeffrey A.
Published: May 2023

Background

Cleft lip and palate is the most common congenital defect of the head and neck, occurring in 1 of 700 live births. Diagnosis often occurs in utero by conventional or 3-dimensional ultrasound. Early cleft lip repair (ECLR) (<3 months of life) for unilateral cleft lip (UCL), regardless of cleft width, has been the mainstay of lip reconstruction at Children's Hospital Los Angeles since 2015. Historically, traditional lip repair (TLR) was performed at 3 to 6 months of life ± preoperative nasoalveolar molding (NAM). Previous publications highlight the benefits of ECLR, such as enhanced aesthetic outcomes, decreased revision rate, better weight gain, increased alveolar cleft approximation, cost savings of NAM, and improved parent satisfaction. Occasionally, parents are referred for prenatal consultations to discuss ECLR. This study evaluates timing of cleft diagnosis, preoperative surgical consultation, and referral patterns to validate whether prenatal diagnosis and prenatal consultation lead to ECLR.

Methods

Retrospective review evaluated patients who underwent ECLR versus TLR ± NAM from 2009 to 2020. Timing of repair, cleft diagnosis, and surgical consultation, as well as referral patterns, were abstracted. Inclusion criteria dictated: age < 3 months for ECLR or 3 to 6 months for TLR, no major comorbidities, and diagnosis of UCL without palatal involvement. Patients with bilateral cleft lip or craniofacial syndromes were excluded.

Results

Of 107 patients, 51 (47.7%) underwent ECLR whereas 56 underwent TLR (52.3%). Average age at surgery was 31.8 days of life for the ECLR cohort and 112 days of life for the TLR cohort. Furthermore, 70.1% of patients were diagnosed prenatally, yet only 5.6% of families had prenatal consults for lip repair, 100% of which underwent ECLR. Most patients were referred by pediatricians (72.9%). Significance was identified between incidence of prenatal consults and ECLR (P = 0.008). In addition, prenatal diagnosis was significantly correlated with incidence of ECLR (P = 0.027).

Conclusions

Our data demonstrate significance between prenatal diagnosis of UCL and prenatal surgical consultation with incidence of ECLR. Accordingly, we advocate for education to referring providers about ECLR and the potential for prenatal surgical consultation in the hopes that families may enjoy the myriad benefits of ECLR.
Outcomes of Local Versus Free Flaps for Reconstruction of the Proximal One-Third of the Leg
Authors: Kondra Katelyn , Roohani Idean , Carey Joseph N.
Published: May 2023

Background

Reconstruction of the proximal one-third of the leg often requires soft tissue transfer to facilitate limb salvage. Tissue transfers are usually local or free flaps depending on wound dimensions, location, and surgeon preference. Historically, the proximal third of the leg was covered with pedicle flaps, but recently, we have used more free flaps in this position. Using data from a level 1 trauma center, we sought to evaluate outcomes of surgical management of proximal-third leg reconstruction across local and free flaps.

Methods

This is an institutional review board–approved, retrospective chart review undertaken at LAC + USC Medical Center from 2007 to 2021. Patient history, demographics, flap characteristics, Gustilo-Anderson fracture classification, and outcomes were collected and analyzed in an internal database. Outcomes of interest included flap failure rates, postoperative complications, and long-term ambulatory status.

Results

Among 394 lower extremity flaps placed, 122 flaps involved the proximal-third leg across 102 patients. Average age of patients was 42.8 ± 15.2 years; of note, the free flap cohort was significantly younger than the local flap cohort (P = 0.019). Ten local flaps suffered from infectious complications: osteomyelitis (n = 6) and hardware infection (n = 4), versus only 1 free flap that suffered from hardware infection; notably, these differences were not significant across cohorts. Free flaps had significantly more flap revisions (13.3%; P = 0.039) and overall flap complications (20.0%; P = 0.031) compared with local flaps; however, partial flap necrosis (4.9%) and flap loss (3.3%) were not significantly different across cohorts. Overall flap survival was 96.7%, and full ambulation was achieved in 42.2% of patients without significant differences across cohorts.

Conclusions

Our evaluation of proximal-third leg wounds demonstrates fewer infectious outcomes with free flaps compared with local flaps. There are multiple confounding variables; however, this finding may speak to the reliability of a robust free flap. Overall, there was no significant difference in patient comorbidities across flap cohorts with great overall flap survival. Ultimately, flap selection did not affect rates of flap necrosis, flap loss, or final ambulatory status.
Evaluation of the Quality of Peri-implantitis Videos on YouTube
Authors: Bariş Kubilay , Karşiyaka Hendek Meltem , Olgun Ebru
Published: May 2023

Although many studies have evaluated the quality of YouTube videos related to dentistry, just one study has evaluated the quality of YouTube videos related to peri-implantitis. The aim of the cross-sectional study was to evaluate the quality of peri-implantitis–related YouTube videos. Two periodontists evaluated 47 videos that met the inclusion criteria, such as the country of upload, source of videos, number of views, likes and dislikes, viewing rate, interaction index, number of days since upload, duration of the video, usefulness score, global quality scale score, and comments. Peri-implantitis was evaluated using a 7-question system of the videos, 44.7% and 55.3% were uploaded by commercial companies and health care professionals, respectively. Although the usefulness score of the videos uploaded by health care professionals was statistically significantly higher (P=0.022), the number of views, numbers of likes and dislikes were similar between the groups (P>0.050). Although the usefulness score and global quality scale score of the perfect videos were statistically different between the groups (P<0.001; P<0.001, respectively), the number of views, numbers of likes, and dislikes were similar. A strong positive correlation was found between the number of views and the number of likes (P˂0.001). A strong negative correlation was found between the interaction index and the number of days since upload (P˂0.001). As a result, YouTube videos on peri-implantitis were limited in number and had poor quality. Thus, videos of perfect quality should be uploaded.
HIV and Hepatitis C Virus-Related Disparities in Undergoing Emergency General Surgical Procedures in the United States, 2016-2019
Authors: Himmelstein Kathryn E.W. , Afif Iman N. , Beard Jessica H. , Tsai Alexander C.
Published: May 2023

Objective:

To determine whether people living with HIV (PLWHIV) and people living with HCV (PLWHCV) experience inequities in receipt of emergency general surgery (EGS) care.

Summary Background Data:

PLWHIV and PLWHCV face discrimination in many domains; it is unknown whether this extends to receipt of EGS care.

Methods:

Using data from the 2016-2019 National Inpatient Sample, we examined 507,458 non-elective admissions of adults with indications for one of the seven highest-burden EGS procedures (partial colectomy, small-bowel resection, cholecystectomy, operative management of peptic ulcer disease, lysis of peritoneal adhesions, appendectomy, or laparotomy). Using logistic regression, we evaluated the association between HIV/HCV status and likelihood of undergoing one of these procedures, adjusting for demographic factors, comorbidities, and hospital characteristics. We also stratified analyses for the seven procedures separately.

Results:

After adjustment for covariates, PLWHIV had lower odds of undergoing an indicated EGS procedure (adjusted odds ratio [aOR], 0.81; 95% confidence interval [CI], 0.73-0.89), as did PLWHCV (aOR, 0.66; 95% CI, 0.63-0.70). PLWHIV had reduced odds of undergoing cholecystectomy (aOR, 0.68; 95% CI, 0.58-0.80). PLWHCV had lower odds of undergoing cholecystectomy (aOR, 0.57; 95% CI, 0.53-0.62) or appendectomy (aOR, 0.76; 95% CI, 0.59-0.98).

Conclusions:

People living with HIV and HCV are less likely than otherwise similar patients to undergo EGS procedures. Further efforts are warranted to ensure equitable access to EGS care for PLWHIV and PLWHCV.
Going for Broke
Authors: Drezdzon Melissa K , Cowley Norah J , Sweeney Daniel P , Peterson Carrie Y , Ridolfi Timothy J , Ludwig Kirk A , Evans Douglas B , Calata Jed F
Published: May 2023

Objective:

Provide a direct comparison of first-year general surgery resident stipends across states and major cities, using the Cost-of-Living Index (COLI) to determine stipend value.

Summary of Background Data:

Financial challenges are among residents’ top sources of stress, and this may be exacerbated in areas with high costs of living. A 2021 survey found that the mean first-year medical resident stipend increased by 0.6%, or $358, from 2020 to 2021 and only 33% of institutions used cost-of-living to determine annual resident stipend adjustments.

Methods:

An AMA database was used to identify accredited general surgery residency programs. 2021-2022 stipend data for first-year general surgery positions was obtained, then data was grouped by state and major city and averaged. Major cities were defined as cities with greater than four programs.

A direct comparison of stipends was performed using the COLI.

Results:

Stipend data was available for 337 of 346 general surgery programs. The national average first-year residency stipend was $60,064±$4,233. The average COLI-adjusted stipend was $57,090±$5,742, with a value loss of -$3,493, or 5%.

For major cities, the average stipend was $63,383±$4,524 and average COLI-adjusted stipend was $46,929±$8,383, with an average value loss of -$16,454, or 26%.

Conclusion:

The financial burdens that residents face cannot be overlooked, and cost of living has a meaningful impact on resident stipend value. Current GME compensation structure limits federal and institutional capacity to adjust for cost of living and creates an insulated market in which residents are under-compensated.
Removal of AMR plasmids using a mobile, broad host-range CRISPR-Cas9 delivery tool
Authors: Sünderhauf David , Klümper Uli , Pursey Elizabeth , Westra Edze R. , Gaze William H. , van Houte Stineke
Published: May 2023

Graphical Abstract

A broad host-range CRISPR-Cas9 delivery tool was developed and used to conjugatively remove antimicrobial resistance (AMR) plasmids (top) and protect a series of coliform and Pseudomonas isolates from AMR plasmid uptake (bottom).
HIV VIRAL SUPPRESSION AND ASSOCIATED FACTORS AMONG CHILDREN AND ADOLESCENTS ON A DOLUTEGRAVIR (DTG) BASED ANTIRETROVIRAL REGIMEN IN TANZANIA MAINLAND
Authors: Maghembe Abdallah Abtwalibe , Boer Marion Sumari-de , Marikias Godbless , Amour Caroline , Mahande Michael Johnson
Published: May 2023

Introduction

Antiretroviral therapy (ART) reduces morbidity and mortality due to human immunodeficiency virus (HIV) infection. The complexity and time-consuming processes, particularly in drug approvals, have contributed to a major challenge to the ongoing success of antiretroviral treatment programs among children and adolescents. In 2019, Tanzania adopted DTG as a first-, second-line and third-line treatment for CALHIV on ART after being approved by the World Health Organisation (WHO). DTG treatment has highly potent antiviral activity, a high genetic barrier to resistance, and a high safety profile. This study aimed to determine HIV viral suppression and associated factors among CALHIV on DTG-based ART in Tanzania Mainland.

Methods

This was a retrospective cohort analysis among children and adolescents living with HIV who were on a DTG-based regimen in Tanzania Mainland between 2019 and 2021. The study utilized routinely collected data from Tanzania Care and Treatment Centres (CTC). We analysed data using STATA version 15 software. We calculated the prevalence of viral suppression by taking the number of children and adolescents with <1000 copies/ml overall study participants. A mixed effect generalized linear model with Poisson distribution and log link function with robust estimator determined the factors associated with HIV viral suppression on a DTG-based regimen.

Results

A total of 63,453 CALHIV on a DTG-based regimen were analysed. The proportion of viral suppression was 91.64%. Overall, 66.19% of previously unsuppressed individuals became suppressed and 88.45% of previously suppressed remained suppressed. Factors leading to lower chances of viral suppression were age 10-14 years (aRR: 0.98; 95%CI: 0.97-0.99), previously unsuppressed prior to starting DTG (aRR: 0.92; 95%CI: 0.91-0.93), duration on ART more than 24 months (aRR: 0.96; 95%CI: 0.94-0.97), not retained in care (aRR: 0.83; 95% CI: 0.77-0.89), severe malnutrition (aRR:0.77; 95%CI: 0.69-0.94) and coastal zone (aRR: 0.98; 95% CI: 0.96-0.99), while those in WHO stage I (aRR: 1.03; 95%CI: 1.01-1.04) and ever received a multi-month prescription (aRR: 1.25; 95% CI: 1.23-1.28) had a higher chance of viral suppression.

Conclusions

The findings support the broad use of DTG-based regimens for eligible CALHIV. Especially those in baseline WHO stage I and those who received the multi-month prescriptions were more likely to achieve viral load suppression. Programs should improve strategies to maintain CALHIV retention in care with interventions like the promotion of teen clubs and teams.
Proteomic Specificity of Soft Contact Lenses for Tear Protein Sampling
Authors: Roden Robert K. , Jiang Fangfang , Zuniga Nathan , Nitz Alyssa , Burlett Rebecca S. , Wright Joshua C. , Shelton Caleb , Reed Alex , Latham Samantha , Roper Connor O. , Patil Leena M. , Ackroyd P. Christine , Payne Samuel H. , Price John C. , Christensen Kenneth A.
Published: May 2023

Soft contact lenses (SCLs) have recently been introduced as an alternative method for human tear protein sampling. However, SCLs are available in a variety of chemical compositions which affect protein binding specificity. Here we analyzed 8 different SCL materials to identify an optimal lens for tear protein sampling. Polymer contamination, mass spectrometry (MS) sample preparation method, total protein capture, individual protein specificity, and SCL cost were all assessed. Using a filter-aided sample prep (FASP) method with 4M guanidine for protein removal, only etafilcon A and verofilcon A did not have significant polymer contamination. Polymer was successfully removed using phosphate buffered saline (PBS) with S-Trap columns for all SCL materials, though yielding a slightly lower number of protein identifications per sample. Minor quantitative differences were observed between SCL materials. However, we also saw significant intersubject variation in protein abundance. Of all the assessed SCL materials, verofilcon A lenses yielded the most total protein while comfilcon A and senofilcon A had the least protein variability. As a newly released daily disposable modality (Precision 1, Alcon), verofilcon A has one of the longest predictable production schedules and one of the lowest costs per lens, making it beneficial for large-scale experiments and diagnostics. Furthermore, we demonstrate how protein binding bias with SCL tear sampling is useful for intra-experiment normalization. Overall, these experiments have led us to optimize our previous protocol for SCL tear protein sampling, highlighting important differences between SCL materials and identifying etafilcon A and verofilcon A as optimal materials for tear protein sampling.

Abstract Figure
Inferior vena cava ultrasoundversuspassive leg raising test in guiding fluid administration in surgical patients prior to spinal anaesthesia: a post-hoc analysis of the ProCRHYSA randomized trial
Authors: Ceruti Samuele , Glotta Andrea , Favre Mathieu , Tasciotti Edoardo , Bona Giovanni , Petrusic Antonietta , Borgeat Alain , Aguirre José , Saporito Andrea
Published: May 2023

Background

Spinal anaesthesia is commonly used for many surgical procedures. One of its potential complications is arterial hypotension, which is nowadays prevented by an empirical fluid administration without any hemodynamic status assessment. However, this practice could increase the risk of volume overload in cardiovascular high-risk patients. Two non-invasive tests are performed to identify fluid-responsiveness: the Inferior Vena Cava Ultrasound (IVCUS) and the Passive Leg Raising Test (PLRT). Aim of this post-hoc analysis was to compare these two methods in spontaneous-breathing patients to assess fluid responsiveness before spinal anaesthesia. Primary outcome was to analyze the incidence of arterial hypotension after spinal anaesthesia in elective surgery patients. Secondary endpoints compared the total fluids amount, the vasoactive drugs administered and the time needed to accomplish the whole procedure in both groups.

Results

The patients analyzed were 132 in the IVCUS group and 148 in the PLRT group; 39.6% of all patients developed arterial hypotension after spinal anaesthesia, 34.8% in the IVCUS group and 43.9% in the PLRT group (Chi-square 2.39, df = 1, p = 0.77). The mean total fluids amount was 794 ± 592 ml; 925 ± 631 ml for IVCUS group and 678 ± 529 ml for PLRT group (p < 0.001). Patients needed vasoactive drugs to restore normal arterial pressure were 18.2% of total, 15% in the IVCUS group and 20% in the PLRT group (p = 0.136). The mean time required to complete the entire procedure was 52 ± 18 min, 48 ± 10 min in the IVCUS group and 56 ± 13 min in the PLRT group (p < 0.001). Complications or out of protocol treatment were registered in 4.6% patients.

Conclusions

Fluid responsiveness assessment in spontaneous breathing patients before spinal anaesthesia could potentially prevent the risk of post-spinal hypotension. In elective surgery, IVCUS could be an accurate method to guide fluid administration in patients undergoing spinal anaesthesia, reducing the incidence of post-spinal hypotension when compared to PLRT.
Dissecting Symptom-linked Dimensions of Resting-State Electroencephalographic Functional Connectivity in Autism with Contrastive Learning
Authors: Tong Xiaoyu , Xie Hua , Fonzo Gregory A. , Zhao Kanhao , Satterthwaite Theodore D. , Carlisle Nancy , Zhang Yu
Published: May 2023

Autism spectrum disorder (ASD) is a common neurodevelopmental disorder characterized bysocial interaction deficits, communication difficulties, andrestricted/repetitive behaviors or fixated interests. Despite its high prevalence, development of effective therapy for ASD is hindered by its symptomatic and neurophysiological heterogeneities. To collectively dissect the ASD heterogeneity in neurophysiology and symptoms, we develop a new analytical framework combining contrastive learning and sparse canonical correlation analysis to identify resting-state EEG connectivity dimensions linked to ASD behavioral symptoms within 392 ASD samples. Two dimensions are successfully identified, showing significant correlations with social/communication deficits (r = 0.70) and restricted/repetitive behaviors (r = 0.45), respectively. We confirm the robustness of these dimensions through cross-validation and further demonstrate their generalizability using an independent dataset of 223 ASD samples. Our results reveal that the right inferior parietal lobe is the core region displaying EEG activity associated with restricted/repetitive behaviors, and functional connectivity between the left angular gyrus and the right middle temporal gyrus is a promising biomarker of social/communication deficits. Overall, these findings provide a promising avenue to parse ASD heterogeneity with high clinical translatability, paving the way for treatment development and precision medicine for ASD.
Dorsal and median raphe neuronal firing dynamics characterized by non-linear metrics
Authors: Rognoni Claudia Pascovich , Serantes Diego , Rodriguez Alejo , Mateos Diego , González Joaqúın , Gallo Diego , Rivas Mayda , Devera Andrea , Lagos Patricia , Rubido Nicolás , Torterolo Pablo
Published: May 2023

The dorsal (DRN) and median (MRN) raphe are the main serotonergic nuclei, being implicated in sleep and mood regulation. The DRN is mainly serotonergic, where neurons have regular spiking activity, slow firing rate (FR), and long action potential duration (APD). The MRN is divided in a median serotonergic region and a paramedian region, containing principally GABAergic neurons, resulting in more diverse neurochemical and electrophysiological features. In the present study, we aimed to enrich the characterization of the raphe nuclei neurons by using non-linear metrics. This was done by analyzing the neuronal basal firing profile in both nuclei of urethane-anesthetized rats using Ordinal Patterns (OP) Entropy, Bins Entropy, and Permutation Lempel-Ziv Complexity (PLZC). In a first step, we found that typical linear metrics – such as FR, coefficient of variation (CV), and APD – fail to distinguish between MRN and DRN neurons, while OP entropy is significantly different between these nuclei. We also found that the FR has a strong linear relationship with CV, Bins Entropy, and PLZC. Similarly, CV has a strong correlation with FR and Bins Entropy, whereas PLZC shows a strong linear fit with Bins Entropy. However, OP Entropy has either a weak or no linear relationship with the rest of the metrics tested, suggesting that OP Entropy is a good metric to differentiate neuronal firing profiles. In a second step, we studied how these metrics are affected by the oscillatory properties of the firing patterns. We found that all metrics are sensitive to rhythmicity – with the exception of OP Entropy. Again, this highlights OP Entropy as a powerful and useful quantity for the characterization of neuronal discharge patterns.
Triphasic production of IFNγ by innate and adaptive lymphocytes following influenza A virus infection
Authors: Finney George E , Hargrave Kerrie E , Pingen Marieke , Purnell Thomas , Todd David , MacDonald Freya , Worrell Julie C , MacLeod Megan KL
Published: May 2023

Interferon gamma (IFNγ) is a potent antiviral cytokine that can be produced by many innate and adaptive immune cells during infection. Currently, our understanding of which cells produce IFNγand where they are located at different stages of an infection are limited. We have used reporter mice to investigatein vivoexpression of IFNγin the lung and secondary lymphoid organs during and following influenza A virus (IAV) infection. We observed a triphasic production of IFNγexpression. Unconventional T cells and innate lymphoid cells, particularly NK cells, were the dominant producers of early IFNγ, while CD4 and CD8 T cells were the main producers by day 10 post-infection. Following viral clearance, some memory CD4 and CD8 T cells continued to produce IFNγin the lungs and draining lymph node. Interestingly, IFNγproduction by lymph node Natural Killer (NK), NKT and innate lymphoid 1 cells also continued to be above naïve levels, suggesting memory-like phenotypes for these cells. Analysis of the localisation of IFNγ+ memory CD4 and CD8 T cells demonstrated that cytokine+ T cells were located near airways and in the lung parenchyma. Following a second IAV challenge, lung IAV specific CD8 T cells rapidly increased their expression of IFNγwhile CD4 T cells in the draining lymph node increased their IFNγresponse. Together, these data suggest that IFNγproduction fluctuates based on cellular source and location, both of which could impact subsequent immune responses.
Pharmaceutical agent cetylpyridinium chloride inhibits immune mast cell function by interfering with calcium mobilization
Authors: Obeng Bright , Potts Christian M. , West Bailey E. , Burnell John E. , Fleming Patrick J. , Shim Juyoung K. , Kinney Marissa S. , Ledue Emily L. , Sangroula Suraj , Baez Vazquez Alan Y. , Gosse Julie A.
Published: May 2023

Cetylpyridinium chloride (CPC) is an antimicrobial used in numerous personal care and janitorial products and food for human consumption at millimolar concentrations. Minimal information exists on the eukaryotic toxicology of CPC. We have investigated the effects of CPC on signal transduction of the immune cell type mast cells. Here, we show that CPC inhibits the mast cell function degranulation with antigen dose-dependence and at non-cytotoxic doses ∼1000-fold lower than concentrations in consumer products. Previously we showed that CPC disrupts phosphatidylinositol 4,5-bisphosphate, a signaling lipid critical for store-operated Ca2+entry (SOCE), which mediates degranulation. Our results indicate that CPC inhibits antigen-stimulated SOCE: CPC restricts Ca2+efflux from endoplasmic reticulum, reduces Ca2+uptake into mitochondria, and dampens Ca2+flow through plasma membrane channels. While inhibition of Ca2+channel function can be caused by alteration of plasma membrane potential (PMP) and cytosolic pH, CPC does not affect PMP or pH. Inhibition of SOCE is known to depress microtubule polymerization, and here we show that CPC indeed dose-dependently shuts down formation of microtubule tracks.In vitrodata reveal that CPC inhibition of microtubules is not due to direct CPC interference with tubulin. In summary, CPC is a signaling toxicant that targets Ca2+mobilization.
Combinatorial immune refocusing within the influenza hemagglutinin head elicits cross-neutralizing antibody responses
Authors: Dosey Annie , Ellis Daniel , Boyoglu-Barnum Seyhan , Syeda Hubza , Saunders Mason , Watson Michael , Kraft John C. , Pham Minh N. , Guttman Miklos , Lee Kelly K. , Kanekiyo Masaru , King Neil P.
Published: May 2023

The head domain of influenza hemagglutinin (HA) elicits potently neutralizing yet mostly strain-specific antibodies during infection and vaccination. Here we evaluated a series of immunogens that combined several immunofocusing techniques for their ability to enhance the functional breadth of vaccine-elicited immune responses. We designed a series of “trihead” nanoparticle immunogens that display native-like closed trimeric heads from the HAs of several H1N1 influenza viruses, including hyperglycosylated variants and hypervariable variants that incorporate natural and designed sequence diversity at key positions in the periphery of the receptor binding site (RBS). Nanoparticle immunogens displaying triheads or hyperglycosylated triheads elicited higher HAI and neutralizing activity against vaccine-matched and -mismatched H1 viruses than corresponding immunogens lacking either trimer-stabilizing mutations or hyperglycosylation, indicating that both of these engineering strategies contributed to improved immunogenicity. By contrast, mosaic nanoparticle display and antigen hypervariation did not significantly alter the magnitude or breadth of vaccine-elicited antibodies. Serum competition assays and electron microscopy polyclonal epitope mapping revealed that the trihead immunogens, especially when hyperglycosylated, elicited a high proportion of antibodies targeting the RBS, as well as cross-reactive antibodies targeting a conserved epitope on the side of the head. Our results yield important insights into antibody responses against the HA head and the ability of several structure-based immunofocusing techniques to influence vaccine-elicited antibody responses.

HIGHLIGHTS

Generalization of trihead antigen platform to several H1 hemagglutinins, including hyperglycosylated and hypervariable variants

Trimer-stabilizing mutations in trihead nanoparticle immunogens lead to lower levels of non-neutralizing antibody responses in both mice and rabbits

Hyperglycosylated triheads elicit higher antibody responses against broadly neutralizing epitopes
Ouvrai: Opening access to remote VR studies of human behavioral neuroscience
Authors: Cesanek Evan , Shivkumar Sabyasachi , Ingram James N. , Wolpert Daniel M.
Published: May 2023

Modern virtual reality (VR) devices offer 6 degree-of-freedom kinematic data with high spatial and tem-poral resolution, making them powerful tools for research on sensorimotor and cognitive functions. We introduce Ouvrai, an open-source solution that facilitates the design and execution of remote VR studies, capitalizing on the surge in VR headset ownership. This tool allows researchers to develop sophisticated experiments using cutting-edge web technologies like the WebXR Device API for browser-based VR, with-out compromising on experimental design. Ouvrai’s features include easy installation, intuitive JavaScript templates, a component library managing front- and back-end processes, and a streamlined workflow. It also integrates APIs for Firebase, Prolific, and Amazon Mechanical Turk and provides data processing utilities for analysis. Unlike other tools, Ouvrai remains free, with researchers managing their web hosting and cloud database via personal Firebase accounts. Through three distinct motor learning experiments, we confirm Ouvrai’s efficiency and viability for conducting remote VR studies.
Cancer driver genes and opportunities for precision oncology revealed by whole genome sequencing 10,478 cancers
Authors: Kinnersley Ben , Sud Amit , Everall Andrew , Cornish Alex J. , Chubb Daniel , Culliford Richard , Gruber Andreas , Lärkeryd Adrian , Mitsopoulos Costas ,   Genomics England Research Consortium, Wedge David , Houlston Richard
Published: May 2023

Identifying cancer driver genes is key for delivering the vision of precision oncology. The falling cost of whole genome sequencing (WGS) potentially makes WGS an attractive single all-encompassing test to identify cancer drivers in a patient, which may not be captured by standard panel testing but are targetable by small molecules. We analysed WGS data on 10,478 patients spanning 35 cancer types recruited to the UK 100,000 Genomes Project. We identified 330 driver genes, including 74 which are novel to any cancer. Across all cancer types 16% of the patients would be eligible for a currently approved therapy. Computational chemogenomic analysis of cancer mutations identified 96 additional targets of compounds that are potentially active and represent candidates for future clinical trials, expanding opportunities for improved patient care.
Concomitant effects of orchiectomy and intermittent hypoxia on hepatic oxidative stress, expression of flavin-containing monooxygenases and transcriptomic profile in mice
Authors: Ganouna-Cohen Gauthier , Marcouiller François , Beauparlant Charles Joly , Droit Arnaud , Belaidi Elise , Bairam Aida , Joseph Vincent
Published: May 2023

Intermittent hypoxia induces oxidative stress and alters hepatic metabolism, likely underlying the association of sleep apnea with non-alcoholic fatty liver disease. In male patients with sleep apnea, metabolic or liver diseases, the levels of testosterone are reduced, and in patients with metabolic diseases, low levels of testosterone are associated with oxidative stress. To assess potential interactions between testosterone and IH on hepatic oxidative stress we used sham-operated or orchiectomized (ORX) mice exposed to normoxia (Nx) or IH (6% O2, 12 cycles/h, 12h/day) for 2 weeks. The activity of prooxidant (NADPH oxidase – NOX), antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase – SOD, Cat, GPx), lipid peroxidation (MDA concentration) and the total concentration of glutathione (GSH) were measured in liver. IH induced a prooxidant profile of enzyme activity (lower SOD activity and higher NOX/SOD and NOX/Cat activity ratio) without altering hepatic MDA and GSH content. Using RNA sequencing followed by a pathway enrichment analysis we identified putative hepatic genes underlying the interactions between IH and testosterone. ORX and IH altered the expression of genes involved in oxidoreductase activities, cytochromes dependent pathways, and glutathione metabolism. Among the genes upregulated in ORX-IH mice, the flavin-containing monooxygenases (FMO) are particularly relevant since these are potent hepatic antioxidant that could help prevent overt oxidative stress in ORX-IH mice.

Graphical Abstract
Serine synthesis pathway upregulated by E-cadherin is essential for the proliferation and metastasis of breast cancers
Authors: Lee Geonhui , Wong Claudia , Cho Anna , West Junior J. , Crawford Ashleigh J. , Russo Gabriella C. , Si Bishwa Ranjan , Kim Jungwoo , Hoffner Lauren , Jang Cholsoon , Jung Moonjung , Leone Robert D. , Konstantopoulos Konstantinos , Ewald Andrew J. , Wirtz Denis , Jeong Sangmoo
Published: May 2023

The loss of E-cadherin (E-cad), an epithelial cell adhesion molecule, has been implicated in the epithelial-mesenchymal transition (EMT), promoting invasion and migration of cancer cells and, consequently, metastasis. However, recent studies have demonstrated that E-cad supports the survival and proliferation of metastatic cancer cells, suggesting that our understanding of E-cad in metastasis is far from comprehensive. Here, we report that E-cad upregulates thede novoserine synthesis pathway (SSP) in breast cancer cells. The SSP provides metabolic precursors for biosynthesis and resistance to oxidative stress, critically beneficial for E-cad-positive breast cancer cells to achieve faster tumor growth and more metastases. Inhibition of PHGDH, a rate-limiting enzyme in the SSP, significantly and specifically hampered the proliferation of E-cad-positive breast cancer cells and rendered them vulnerable to oxidative stress, inhibiting their metastatic potential. Our findings reveal that E-cad adhesion molecule significantly reprograms cellular metabolism, promoting tumor growth and metastasis of breast cancers.
Comparative electric and ultrastructural studies of cable bacteria reveal new components of conduction machinery
Authors: Digel Leonid , Justesen Mads L. , Bonné Robin , Fransaert Nico , Wouters Koen , Jensen Pia B. , Plum-Jensen Lea E. , Marshall Ian P. G. , Nicolas-Asselineau Louison , Drace Taner , Bøggild Andreas , Hansen John L. , Schramm Andreas , Bøjesen Espen D. , Nielsen Lars P. , Manca Jean V. , Boesen Thomas
Published: May 2023

Cable bacteria encompass at least two genera, and they are known to vary greatly in habitat preferences and filament thickness. We systematically investigated variations and similarities in cellular structures and electrical properties of different cable bacteria strains. Using SEM, TEM, STEM-EDX and ToF-SIMS, we characterized shared features of cable bacteria, such as inner and outer membranes, surface layer and cell junction architecture, as well as strain specific features, like the number and size of periplasmic conductive fibers (PCFs). Our data indicates that the PCFs are organized as loose stranded rope-like structures. With spatially resolved elemental analysis we detected nickel-containing co-factors within the PCF of cable bacteria strains in both genera suggesting a conserved conduction mechanism. Electrical conductivity of different cable bacteria strains showed a range of values covering three orders of magnitude indicating an unknown metabolic adaptation. Using cryogenic electron tomography we discovered multiple polar chemosensory arrays, abundant cytoplasmic inner membrane-attached vesicles (IMVs), polysomes and inner membrane invaginations that shed light on cable bacteria metabolism including complex motility control mechanisms, localized protein synthesis, and membrane remodeling. We propose that the IMVs discovered in this work are novel metabolic hubs closely connected to the unique conductive fiber structure of cable bacteria.
Pupil size predicts the onset of exploration in brain and behavior
Authors: Shourkeshti Akram , Marrocco Gabriel , Jurewicz Katarzyna , Moore Tirin , Ebitz R. Becket
Published: May 2023

In uncertain environments, intelligent decision-makers exploit actions that have been rewarding in the past, but also explore actions that could be even better. Several neuromodulatory systems are implicated in exploration, based, in part, on work linking exploration to pupil size–a peripheral correlate of neuromodulatory tone and index of arousal. However, pupil size could instead track variables that make exploration more likely, like volatility or reward, without directly predicting either exploration or its neural bases. Here, we simultaneously measured pupil size, exploration, and neural population activity in the prefrontal cortex while two rhesus macaques explored and exploited in a dynamic environment. We found that pupil size under constant luminance specifically predicted the onset of exploration, beyond what could be explained by reward history. Pupil size also predicted disorganized patterns of prefrontal neural activity at both the single neuron and population levels, even within periods of exploitation. Ultimately, our results support a model in which pupil-linked mechanisms promote the onset of exploration via driving the prefrontal cortex through a critical tipping point where prefrontal control dynamics become disorganized and exploratory decisions are possible.
Chaperone-driven entropic separation of amyloid nanofilament bundles
Authors: Vilar Jose M. G. , Rubi J. Miguel , Saiz Leonor
Published: May 2023

The disassembly of misfolded protein aggregates is a requirement for the proper functioning of cells. It has implications in multiple neuropathologies, such as Alzheimer’s and Parkinson’s diseases. The active unbundling of fibrillar aggregates has recently been identified as a key, rate-limiting step in the disassembly process. Yet, the nature of the underlying molecular mechanism remains an outstanding question. Here, we develop a coarse-grained computational approach from the atomistic structural information and show that the interactions of molecules tethered to fibrils lead to entropic forces consistent with the unbundling process observed in amyloid α-synuclein disaggregation by Hsp70. We uncover two main types of entropic effects, categorized as intra- and inter-protofilament, which are differentially affected by the system parameters and conditions. Our results show that only highly efficient chaperone systems can overcome the free energy cost of the lateral association between two protofilaments. Through the analysis of cryo-electron tomography and high-speed atomic force microscopy data, we find that co-chaperone networks and ATP hydrolysis are needed to achieve the conditions for highly efficient entropic force generation. We highlight the implications of these results for the design of targeted therapies for the underlying neuropathologies.
Development and validation of a high throughput screening platform to enable target identification in skeletal muscle cells from Duchenne Muscular Dystrophy (DMD) patients
Authors: Hariharan Santosh , Lorintiu Oana , Lee Chia-Chin , Duchemin-Pelletier Eve , Li Xianfeng , Healy Aileen , Doyonnas Regis , Selig Luc , Poydenot Pauline , Ventre Erwann , Weston Andrea , Owens Jane , Christoforou Nicolas
Published: May 2023

Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle degenerating disease caused by dystrophin deficiency. Effective methods for drug discovery for the treatment of DMD requires systems to be physiologically relevant, scalable, and effective. To this end, the Myoscreen platform offers a scalable and physiologically relevant system for generating and characterizing patient-derived myotubes. Morphological profiling is a powerful technique involving the simultaneous measurement of hundreds of morphological parameters from fluorescence microscopy images and using machine learning to predict cellular activity. Here, we describe combining the Myoscreen platform and high dimensional morphological profiling to accurately predict a phenotype associated with the lack of Dystrophin expression in patient derived myotubes. Using this methodology, we evaluated a series of Dystrophin-associated protein complex (DAPC) candidates and identified that the combination of Utrophin and α- Sarcoglycan yielded highest morphological differences between DMD and non-DMD donors. Finally, we validated this methodology by knocking down Dystrophin expression in non-DMD cells as well as introducing Dystrophin expression in DMD cells. Knocking down Dystrophin in non- DMD cells shifted their morphological profile to one that is similar to DMD cells while introducing Dystrophin in DMD cells shifted their morphological profile towards non-DMD cells. In conclusion, we have developed a platform that accurately predicts the DMD disease phenotype in a disease relevant cell type. Ultimately this platform may have wide applications in the drug development process include identification of disease modifier genes, screening of novel therapeutic moieties, and as a potency assay for future therapeutics.
Cone Synaptic function is modulated by the leucine rich repeat (LRR) adhesion molecule LRFN2
Authors: Hasan Nazarul , Gregg Ronald G.
Published: May 2023

Daylight vision is mediated by cone photoreceptors in vertebrates, which synapse with bipolar cells (BCs) and horizontal (HCs) cells. This cone synapse is functionally and anatomically complex, connecting to 8 types of depolarizing (DBC) BCs and 5 types of hyperbolizing BCs (HBCs). The dendrites of DBCs and HCs cells make invaginating ribbon synapses with the cone axon terminal, while HBCs form flat synapses with the cone pedicles. The molecular architecture that underpins this organization is relatively poorly understood. To identify new proteins involved in synapse formation and function we used an unbiased proteomic approach and identified LRFN2 (Leucine-rich repeat and fibronectin III domain-containing 2) as a component of the DBC signaling complex. LRFN2 interacts with TRPM1 and is selectively expressed at cone terminals and co-localizes with PNA, and other DBC signalplex members. In the absence of LRFN2 the cone-mediated photopic electroretinogram b-wave amplitude is reduced. In LRFN2 deficient mice, the synaptic markers: LRIT3, ELFN2, mGluR6, TRPM1 and GPR179 are properly localized. Similarly, LRFN2 expression and localization is not dependent on these synaptic proteins. These data demonstrate that LRFN2 likely interacts with TRPM1 and its absence compromises normal synaptic transmission between cones and cone DBCs cells.

Significance Statement

Signaling between cone photoreceptors and the downstream bipolar cells is critical to normal vision. Cones synapse with 13 different types of bipolar cells forming an invaginating ribbon synapses with 8 types, and flat synapse with 5 types, to form one of the most complex synapses in the brain. In this report a new protein, LRFN2 (Leucine-rich repeat and fibronectin III domain-containing 2), was identified that is expressed on the cones synapses. UsingLrfn2knockout mice we show LRFN2 is required for the normal cone signaling.