Light-activated BioID - an optically activated proximity labeling system to study protein-protein interactions.

Proximity labeling with genetically encoded enzymes is widely used to study protein-protein interactions in cells. However, the accuracy of proximity labeling is limited by a lack of control over the enzymatic labeling process. Here, we present a light-activated proximity labeling technology for mapping protein-protein interactions at the cell membrane with high accuracy and precision. Our technology, called light-activated BioID (LAB), fuses the two halves of the split-TurboID proximity labeling enzyme to the photodimeric proteins CRY2 and CIB1. We demonstrate, in multiple cell lines, that upon illumination with blue light, CRY2 and CIB1 dimerize, reconstitute split-TurboID and initiate biotinylation. Turning off the light leads to the dissociation of CRY2 and CIB1 and halts biotinylation. We benchmark LAB against the widely used TurboID proximity labeling method by measuring the proteome of E-cadherin, an essential cell-cell adhesion protein. We show that LAB can map E-cadherin-binding partners with higher accuracy and significantly fewer false positives than TurboID.

An expanded palette of dopamine sensors for multiplex imaging in vivo.

Genetically encoded dopamine sensors based on green fluorescent protein (GFP) enable high-resolution imaging of dopamine dynamics in behaving animals. However, these GFP-based variants cannot be readily combined with commonly used optical sensors and actuators, due to spectral overlap. We therefore engineered red-shifted variants of dopamine sensors called RdLight1, based on mApple. RdLight1 can be combined with GFP-based sensors with minimal interference and shows high photostability, permitting prolonged continuous imaging. We demonstrate the utility of RdLight1 for receptor-specific pharmacological analysis in cell culture, simultaneous assessment of dopamine release and cell-type-specific neuronal activity and simultaneous subsecond monitoring of multiple neurotransmitters in freely behaving rats. Dual-color photometry revealed that dopamine release in the nucleus accumbens evoked by reward-predictive cues is accompanied by a rapid suppression of glutamate release. By enabling multiplexed imaging of dopamine with other circuit components in vivo, RdLight1 opens avenues for understanding many aspects of dopamine biology.

The financial and employment effects of coronavirus disease 2019 on physicians in the United States.

Although the coronavirus disease 2019 (COVID-19) pandemic has created havoc with the U.S healthcare system and physicians, the financial and contractual implications for physicians are now beginning to come to the forefront. Financial assistance from the federal government has mainly been received by hospitals, which have borne the brunt of the COVID-19 illness. Some physician groups have, or are, receiving assistance through a few programs, although the accelerated and advance payments have been suspended. Employed surgeons are now being furloughed, terminated, or persuaded to agree to a significant cut in pay, forego bonuses, or take leave without pay as healthcare systems and some physician groups have started to experience the consequences of halting elective procedures. Newly hired surgeons might be forced in a few cases to agree to delays in starting their employment, new amendments, changes in employment status, and other terms for fear of losing their employment. In the present report, we have explained some agreement terminology and options available to allow physicians to understand the terms of their employment agreement and make their decisions after consulting with an expert healthcare attorney.

Adopting evidence-based medically assisted treatments in substance abuse treatment organizations: roles of leadership socialization and funding streams.

This paper examines the organizational adoption of medically assisted treatments (MAT) for substance use disorders (SUDs) in a representative sample of 555 US for-profit and not-for-profit treatment centers. The study examines organizational adoption of these treatments in an institutionally contested environment that traditionally has valued behavioral treatment, using sociological and resource dependence frameworks. The findings indicate that socialization of leadership, measured by formal clinical education, is related to the adoption of MAT. Funding patterns also affect innovation adoption, with greater adoption associated with higher proportions of earned income from third party fees for services, and less adoption associated with funding from criminal justice sources. These findings may generalize to other social mission-oriented organizations where innovation adoption may be linked to private and public benefit values inherent in the type of socialization of leadership and different patterns of funding support.

CD8+ T cells are necessary for improved sepsis survival induced by CD28 agonism in immunologically experienced mice.

Introduction: A hallmark of T cell dysregulation during sepsis is the downregulation of costimulatory molecules. CD28 is one of T cell costimulatory molecules significantly altered on memory T cells during sepsis. We recently showed that treatment with a αCD28 agonist in septic immunologically experienced mice led to improved survival. Therefore, here we aimed to identify the cell subset(s) necessary for the survival benefit observed in the context of CD28 agonism, and to further investigate the mechanism by which CD28 agonism improves sepsis survival in immunologically experienced mice. Methods: Mice received specific pathogen inoculation to generate memory T cell populations similar in frequency to that of adult humans. Once these infections were cleared and the T cell response had transitioned to the memory phase, animals were rendered septic via cecal ligation and puncture in the presence or absence of an agonistic anti-CD28 mAb. Results: Results demonstrated that CD8+ T cells, and not bulk CD4+ T cells or CD25+ regulatory T cells, were necessary for the survival benefit observed in CD28 agonist-treated septic immunologically experienced mice. Upon examination of these CD8+ T cells, we found that CD28 agonism in septic immunologically experienced mice was associated with an increase in Foxp3+ CD8+ T cells as compared to vehicle-treated controls. When CD8+ T cells were depleted in septic immunologically experienced mice in the setting of CD28 agonism, a significant increase in levels of inflammatory cytokines in the blood was observed. Discussion: Taken together, these results indicate that CD28 agonism in immunologically experienced mice effectively suppresses inflammation via a CD8+-dependent mechanism to decrease mortality during sepsis.

Derivation and Validation of Generalized Sepsis-induced Acute Respiratory Failure Phenotypes Among Critically Ill Patients: A Retrospective Study.

Background: Septic patients who develop acute respiratory failure (ARF) requiring mechanical ventilation represent a heterogenous subgroup of critically ill patients with widely variable clinical characteristics. Identifying distinct phenotypes of these patients may reveal insights about the broader heterogeneity in the clinical course of sepsis. We aimed to derive novel phenotypes of sepsis-induced ARF using observational clinical data and investigate their generalizability across multi-ICU specialties, considering multi-organ dynamics. Methods: We performed a multi-center retrospective study of ICU patients with sepsis who required mechanical ventilation for ≥24 hours. Data from two different high-volume academic hospital systems were used as a derivation set with N=3,225 medical ICU (MICU) patients and a validation set with N=848 MICU patients. For the multi-ICU validation, we utilized retrospective data from two surgical ICUs at the same hospitals (N=1,577). Clinical data from 24 hours preceding intubation was used to derive distinct phenotypes using an explainable machine learning-based clustering model interpreted by clinical experts. Results: Four distinct ARF phenotypes were identified: A (severe multi-organ dysfunction (MOD) with a high likelihood of kidney injury and heart failure), B (severe hypoxemic respiratory failure [median P/F=123]), C (mild hypoxia [median P/F=240]), and D (severe MOD with a high likelihood of hepatic injury, coagulopathy, and lactic acidosis). Patients in each phenotype showed differences in clinical course and mortality rates despite similarities in demographics and admission co-morbidities. The phenotypes were reproduced in external validation utilizing an external MICU from second hospital and SICUs from both centers. Kaplan-Meier analysis showed significant difference in 28-day mortality across the phenotypes (p<0.01) and consistent across both centers. The phenotypes demonstrated differences in treatment effects associated with high positive end-expiratory pressure (PEEP) strategy. Conclusion: The phenotypes demonstrated unique patterns of organ injury and differences in clinical outcomes, which may help inform future research and clinical trial design for tailored management strategies.

Understanding 'error' in the forensic sciences: A primer.

This paper distils seven key lessons about 'error' from a collaborative webinar series between practitioners at Victoria Police Forensic Services Department and academics. It aims to provide the common understanding of error necessary to foster interdisciplinary dialogue, collaboration and research. The lessons underscore the inevitability, complexity and subjectivity of error, as well as opportunities for learning and growth. Ultimately, we argue that error can be a potent tool for continuous improvement and accountability, enhancing the reliability of forensic sciences and public trust. It is hoped the shared understanding provided by this paper will support future initiatives and funding for collaborative developments in this vital domain.

Optimizing the Synergistic Potential of Pseudo-Labels from Radiology Notes and Annotated Ground Truth in Identifying Pulmonary Opacities on Chest Radiographs for Early Detection of Acute Respiratory Distress Syndrome.

Acute Respiratory Distress Syndrome (ARDS) is a life-threatening lung injury, hallmarks of which are bilateral radiographic opacities. Studies have shown that early recognition of ARDS could reduce severity and lethal clinical sequela. A Convolutional Neural Network (CNN) model that can identify bilateral pulmonary opacities on chest x-ray (CXR) images can aid early ARDS recognition. Obtaining large datasets with ground truth labels to train CNNs is challenging, as medical image annotation requires clinical expertise and meticulous consideration. In this work, we implement a natural language processing pipeline that extracts pseudo-labels CXR images by parsing radiology notes for abnormal findings. We obtain ground-truth annotations from clinicians for the presence of pulmonary opacities for a subset of these images. A knowledge distillation-based teacher-student training framework is implemented to leverage the larger dataset with noisy pseudo-labels. Our results show an AUC of 0.93 (95%CI 0.92-0.94) for the prediction of bilateral opacities on chest radiographs.

Uncertainty-Aware Convolutional Neural Network for Identifying Bilateral Opacities on Chest X-rays: A Tool to Aid Diagnosis of Acute Respiratory Distress Syndrome.

Acute Respiratory Distress Syndrome (ARDS) is a severe lung injury with high mortality, primarily characterized by bilateral pulmonary opacities on chest radiographs and hypoxemia. In this work, we trained a convolutional neural network (CNN) model that can reliably identify bilateral opacities on routine chest X-ray images of critically ill patients. We propose this model as a tool to generate predictive alerts for possible ARDS cases, enabling early diagnosis. Our team created a unique dataset of 7800 single-view chest-X-ray images labeled for the presence of bilateral or unilateral pulmonary opacities, or 'equivocal' images, by three blinded clinicians. We used a novel training technique that enables the CNN to explicitly predict the 'equivocal' class using an uncertainty-aware label smoothing loss. We achieved an Area under the Receiver Operating Characteristic Curve (AUROC) of 0.82 (95% CI: 0.80, 0.85), a precision of 0.75 (95% CI: 0.73, 0.78), and a sensitivity of 0.76 (95% CI: 0.73, 0.78) on the internal test set while achieving an (AUROC) of 0.84 (95% CI: 0.81, 0.86), a precision of 0.73 (95% CI: 0.63, 0.69), and a sensitivity of 0.73 (95% CI: 0.70, 0.75) on an external validation set. Further, our results show that this approach improves the model calibration and diagnostic odds ratio of the hypothesized alert tool, making it ideal for clinical decision support systems.

Femoral-Femoral Bypass for Limb Salvage to Correct Common Iliac Occlusion after Lower Extremity Trauma.

BACKGROUND: Roadway injuries are a leading cause of lower extremity vascular trauma. Treating these injuries involves controlling life-threatening hemorrhage and restoring distal perfusion. MATERIALS AND METHODS: We describe a unique presentation of chronic iliac artery occlusion in the setting of blunt trauma requiring extra-anatomic bypass for maximal limb salvage. RESULTS: A 50-year-old male presented after a pedestrian versus auto accident. He had mangled bilateral lower extremities and was taken emergently for lower extremity amputations. He was found to have chronic left common iliac occlusion and a femoral-femoral bypass was performed to assist with healing his left below-the-knee amputation. DISCUSSION: Lack of adequate perfusion can cause poor outcomes in limb salvage. This case demonstrated that lower extremity trauma can be complicated by chronic vascular disease. Reperfusion and adequate wound healing can be accomplished by using bypass grafting after more traditional reperfusion techniques fail.

Video-Assisted Thoracoscopic Internal Rib Fixation.

Rib fractures result in serious morbidity and mortality after trauma. Although there is ongoing debate about surgical rib fixation, it is increasingly important for some patients. Minimally invasive techniques for rib fixation are gaining traction within the trauma community. We present an observational experience at our level 1 trauma center with our first 10 cases of video-assisted thoracoscopic surgery (VATS) internal rib fixation. Video-assisted thoracoscopic surgery internal plates are especially helpful for rib fractures under the scapula, which are difficult to access traditionally. This technique is also excellent at reducing complex segmental fractures as the bridge can span across multiple fractures with a single post on either side. They also work well for posterior fractures where multiple screws cannot be placed. Video-assisted thoracoscopic surgery internal rib fixation is a viable and exciting option for surgical fixation. The plates work particularly well for certain fracture patterns.

Latino adolescent mothers and fathers in south Texas managing violence.

During the course of this study young Latino parents living in San Antonio, TX shared their stories. Their stories draw attention to the multiple forms of violence they experienced in the past, continue to experience, and are forced to manage. Their lives occur within the context of enduring social disparities and exposure to violence. At a young age they develop strategies to manage violence, but new challenges with violence manifest themselves as they enter into intimate partner relationships. For some of these youth, parenthood is perceived as a transforming experience that offers an opportunity for a positive change in their life trajectory. They recognize the need to find non-violent ways to manage their relationships with their intimate partner and their children. Culturally-appropriate intervention strategies that focus on developing relationships of equality and balance are needed.

Refining a Church-Based Lifestyle Intervention Targeting African-American Adults at Risk for Cardiometabolic Diseases: A Pilot Study.

OBJECTIVE: The pilot study was intended to test the feasibility of a multiple-component lifestyle intervention targeting African American adults in a weight control and cardiometabolic risk reduction program on diet, activity, and stress, using community-engagement principles. METHODS: Applying mixed qualitative and quantitative measures, the intervention had a two-part sequential study design consisting of 12 weekly small group sessions that provided individual and group counseling in nutrition, exercise, and mindfulness, while incorporating focus group and interactive techniques to learn about barriers and acceptable practices for this population. The program was implemented at an African-American church in Nashville, Tennessee. RESULTS: Thirty-four participants (aged 56.1 ± 11 years, body mass index (BMI) 36.7 ± 6.6 kg/m2) completed the intervention. Lifestyle changes after the 12 weekly sessions showed some positive trends including reduced sodium intake (from 2725.3 ± 326.5 to 2132 ± 330, mg/day, P = 0.008), increased walking steps (from 4392.1 ± 497.2 to 4895.3 ± 497.9, steps/day, not significant), and slightly decreased Perceived Stress Scale (PSS) scores (from 13.7 ± 1.4 to 12.4 ± 1.5, not significant). Body fat % among male participants decreased significantly (from 33.8 ± 2.6 to 28 ± 2.6, %, P = 0.043). Among cardiometabolic risk biomarkers, hemoglobin A1c (HbA1c) decreased significantly (from 6.6 ± 0.2 to 6.1 ± 0.2, %, P < 0.001). The baseline PSS score was positively associated with baseline adiposity levels (e.g., weight, ß = 2.4, P = 0.006). Twenty-one participants took part in focus groups during the program to identify barriers to healthy lifestyle changes. Primary barriers reported were price, time for preparing healthy meals, unfamiliarity with mindfulness activities, their health condition, and daily schedule available for physical activities. CONCLUSIONS: This church-based pilot intervention was proven feasible by showing modest progress in reducing adiposity and decreasing HbA1c levels. The focus group and interactive methods facilitated program direction. Future full-scale studies are warranted to identify key strategies that provide more personalized approaches and supportive environments to sustain a healthy lifestyle among these at risk minorities with limited resources.

Isolation and characterization of an adventitious avian leukosis virus isolated from commercial Marek's disease vaccines.

Commercial Marek's disease (MD) vaccines produced by two manufacturers were tested for possible contamination with avian leukosis virus (ALV). Samples of MD vaccines manufactured by two companies (A and B) were received from a breeder company; samples were also received directly from vaccine company B. Using virus isolation tests, samples initially tested positive for subgroup E (endogenous) ALV. However, upon repassage, the vaccines also tested positive for exogenous ALV. The isolated exogenous ALV proved to be a subgroup A virus, as determined by flow cytometry using polyclonal chicken antibodies specific for various subgroups of ALV, and by DNA sequencing of the envelope glygoprotein (gp85). The exogenous ALV isolated from MD vaccines was inoculated in chickens from ADOL lines 15I(5) x 7(1) and 0 to determine its pathogenicity and compare it with that of Rous-associated-virus-1 (RAV-1), the prototype strain of ALV-A. Each chicken from each line was inoculated with approximately 10,000 infectious units of RAV-1 or the ALV-A isolated from vaccines termed B-39 virus at 7th day of embryonation. At hatch, and at 4, 8, and 16 wk of age, chickens were tested for viremia and cloacal shedding; chickens were also observed for ALV-induced tumors within 16 wk of age. Viremia and cloacal shedding results suggest that chickens from both lines were susceptible to infection with either virus. Within 16 wk of age, the proportion of ALV tumors induced by strain B-39 in line 0 and line 15I5 x 7(1) chickens was 0% and 12%, respectively, compared with 62% and 67% in chickens inoculated with RAV-1. The data indicate that commercial MD vaccines produced by two manufacturers were contaminated with endogenous subgroup E and an exogenous subgroup A ALV. Further, data from biological characterization suggest that the ALV-A isolated from commercial MD vaccines is of low oncogenicity, compared with that of RAV-1. GenBank accession numbers: The gp85 gene sequences of ALV isolated from commercial Marek's disease vaccines have been deposited in GenBank and assigned the following accession numbers: A46 subgroup A, DQ412726 ; B53 subgroup A, DQ412727; A46 subgroup E, DQ412728; B53 subgroup E, DQ412729.

African American women with breast cancer and their spouses' perception of care received from physicians.

Several studies have been conducted which have examined the communication patterns and interpersonal relationships among minority patients and their physicians. These studies suggest that there are differences in the care received from physicians based on the racial and cultural background of the patient. However, little is known about the interpersonal relationships and communication patterns of AA couples coping with a breast cancer diagnosis and their physicians. This report presents the outcomes of a qualitative study undertaken to examine the perceptions of breast cancer care rendered by physicians to African American couples. The AA couples involved in the study reported that the care they received was compassionate, competent, comprehensive and comparable to other women. Each of these qualities was identified by as being essential to quality patient care.

Complications and long-term survival for alcoholic patients with resectable lung cancer.

BACKGROUND: The aim of this study was to determine the surgical risks and long-term survival in alcoholic patients undergoing resection for non-small-cell lung cancer. METHODS: Nineteen resected patients comprising the alcoholic group were identified by either a Diagnostic and Statistical Manual of Mental Disorders-IV diagnosis of alcohol dependence/abuse, or an alcohol consumption of 60 oz/d or more. Alcoholic patients were compared with 37 nonalcoholic patients undergoing resection. RESULTS: Alcoholic patients had an increase in major infectious complications (37% [7 of 19] versus 5% [2 of 37], P = 0.005), respiratory failure (42% [8 of 19] versus 5% [2 of 37], P

Node-link-mapping-enhanced group treatment for pathological gambling.

Two experiments evaluated a group treatment for pathological gambling that used node-link mapping techniques to enhance treatment effectiveness. In Experiment 1, 13 (8 female) pathological gamblers were randomly assigned to either a mapping group (n=4), a nonmapping group (n=4), or a wait-list control group (n=5). The treatments were conducted by Master's level counselors during 90-min sessions conducted twice per week for 8 weeks. Participants were assessed pre- and post-8 weeks and then 6 months later on Diagnostic and Statistical Manual of Mental Disorders 4th ed. (DSM-IV) pathological gambling criteria, three self-ratings of control of gambling, gambling expenditure, and gambling bout duration. Experiment 2 replicated the mapping (n=9; 8 female) and wait-list (n=10; 8 female) conditions of Experiment 1 and expanded the dependent measures to include assessment of changes in cooccurring depression and anxiety. The node-link-mapping-enhanced group treatment produced improvements in more of the dependent measures of pathological gambling than treatment without maps (Experiment 1) or an equivalent-length waiting period (Experiments 1 and 2). It also produced larger decreases in cooccurring depression and anxiety than an equivalent-length waiting period (Experiment 2). The results are consistent with previous treatment research with substance abusers.

The potential role of sports psychology in the obesity epidemic.

Sports psychologists play an important role in enhancing performance among athletes. In conjunction with team physicians, they can also shed light on psychological disorders common in athletes, such as mood and eating disorders, and overtraining syndrome. Sports psychologists can also lend their expertise to assist with injury prevention and recovery and compliance issues. Sports psychology has a role in helping to reverse the growing obesity epidemic among school-aged children. These professionals, working with coaches, can increase children's levels of physical activity. Cognitive-behavioral techniques could lead to enhanced enjoyment, increased participation, improved school performance, and a reduction in obesity.

Chronic Methamphetamine Causes Differential Expression of Immediate Early Genes in the Nucleus Accumbens and Midbrain of Rats.

The present study investigated whether chronic methamphetamine (METH) would suppress METH-induced mRNA expression of immediate early genes (IEGs) in the rat brain. Rats were given METH or saline over two weeks. After an overnight withdrawal, saline- and METH-pretreated rats received an acute saline or METH challenge. The acute METH challenge increased expression of members of activator protein 1 (AP-1) and Nr4a IEG families in the nucleus accumbens (NAc) and midbrain of saline-pretreated rats. Chronic METH exposure attenuated the effects of acute METH challenge on AP-1 IEG expression in the NAc. However, chronic METH failed to attenuate acute METH-induced increases of Nr4a1 and Nr4a3 expression in the NAc. In contrast to observations in the NAc, chronic METH did not prevent acute METH-induced changes in IEG expression in the midbrain. These results suggest that these two brain regions that are implicated in neuroplastic effects of illicit substances might be differentially affected by psychostimulants.

Blood transfusions in critical care: improving safety through technology & process analysis.

A multidisciplinary safety initiative transformed blood transfusion practices at St. Luke's Episcopal Hospital in Houston, Texas. An intense analysis of a mistransfusion using the principles of a Just Culture and the process of Cause Mapping identified system and human performance factors that led to the transfusion error. Multiple initiatives were implemented including technology, education and human behaviour change. The wireless technology of Pyxis Transfusion Verification by CareFusion is effective with the rapid infusion module efficient for use in critical care. Improvements in blood transfusion safety were accomplished by thoroughly evaluating the process of transfusions and by implementing wireless electronic transfusion verification technology. During the 27 months following implementation of the CareFusion Transfusion Verification there have been zero cases of transfusing mismatched blood.