Dangerous organophosphorus (OP) compounds have been used as insecticides in agriculture and in chemical warfare. Because exposure to OP could create a danger for humans in the future, ...butyrylcholinesterase (BChE) has been developed for prophylaxis to these chemicals. Because it is impractical to obtain sufficient quantities of plasma BChE to treat humans exposed to OP agents, the production of recombinant BChE (rBChE) in milk of transgenic animals was investigated. Transgenic mice and goats were generated with human BChE cDNA under control of the goat β-casein promoter. Milk from transgenic animals contained 0.1-5 g/liter of active rBChE. The plasma half-life of PEGylated, goat-derived, purified rBChE in guinea pigs was 7-fold longer than non-PEGylated dimers. The rBChE from transgenic mice was inhibited by nerve agents at a 1:1 molar ratio. Transgenic goats produced active rBChE in milk sufficient for prophylaxis of humans at risk for exposure to OP agents.
The endocannabinoid 2-AG is highly susceptible to its hydrolysis into AA, which activates neutrophils through de novo LTB(4) biosynthesis, independently of CB activation. In this study, we show that ...2-AG and AA stimulate neutrophils to release antimicrobial effectors. Supernatants of neutrophils activated with nanomolar concentrations of 2-AG and AA indeed inhibited the infectivity of HSV-1 and RSV. Additionally, the supernatants of 2-AG- and AA-stimulated neutrophils strongly impaired the growth of Escherichia coli and Staphylococcus aureus. This correlated with the release of a large amount (micrograms) of α-defensins, as well as a limited amount (nanograms) of LL-37. All the effects of AA and 2-AG mentioned above were prevented by inhibiting LTB(4) biosynthesis or by blocking BLT(1). Importantly, neither CB(2) receptor agonists nor antagonists could mimic nor prevent the effects of 2-AG, respectively. In fact, qPCR data show that contaminating eosinophils express ∼100-fold more CB(2) receptor mRNA than purified neutrophils, suggesting that CB(2) receptor expression by human neutrophils is limited and that contaminating eosinophils are likely responsible for the previously documented CB(2) expression by freshly isolated human neutrophils. The rapid conversion of 2-AG to AA and their subsequent metabolism into LTB(4) promote 2-AG and AA as multifunctional activators of neutrophils, mainly exerting their effects by activating the BLT(1). Considering that nanomolar concentrations of AA or 2-AG were sufficient to impair viral infectivity, this suggests potential physiological roles for 2-AG and AA as regulators of host defense in vivo.
Orthopedic oncology surgeons commonly engage in prolonged and complex surgical procedures. These types of surgeries increase the risk of physical and psychological stressors, which may in turn make ...these physicians prone to work-related occupational injuries.
The aim of this study was to explore in orthopedic oncologists, the prevalence of work-related physical injuries and psychological disturbances.
A modified version of the physical discomfort survey was developed to assess occupational injuries among orthopedic oncology surgeon members of the Musculoskeletal Tumor Society, the Canadian Orthopedic Oncology Society and European Musculoskeletal Oncology Societies. The survey was sent by email, and it explored musculoskeletal complaints, psychological disturbances, treatment required for these complaints and the requirement of time off work.
A total of 67 surgeon responses were collected. A high number of orthopedic oncologists (84%) reported an occupational injury. Low back pain (39%) was the most prevalent musculoskeletal condition, followed by lumbar disk herniation (16%), shoulder tendinitis (15%) and lateral epicondylitis (13%). Of the cohort, 46% required surgery and 31% required time off work due to their injury. Thirty-three respondents reported a psychological disorder. Burnout (27%), anxiety (20%) and insomnia (20%) were the most commonly reported. Time required off work due to injury was associated with old age and years in practice.
Orthopedic oncology surgeons report a high prevalence of work-related disorders. Lower back related injury and burnout were the most reported disorders. Improving operative room ergonomics and prevention of stress related to the work environment should be areas to explore in upcoming research.
A 19-year-old woman presented with a primary mediastinal B-cell lymphoma invading the superior vena cava with associated thrombosis of the left brachiocephalic and subclavian vein. She underwent ...thrombolysis followed by chemotherapy. The midtreatment 18F-FDG PET/CT demonstrated important regression of the primary mediastinal B-cell lymphoma, but showed intense focal hepatic uptake in segment IV, without a corresponding lesion on ultrasonography, non-contrast-enhanced low-dose CT, and MRI. This focal uptake disappeared on a subsequent 18F-FDG PET/CT study when the radiotracer was injected in the foot, suggesting an anomalous venous return pathway that persisted despite thrombolysis.
IntroductionThe frail elderly in Canada face a tough decision when they start to lose autonomy: whether to stay at home or move to another location. This study seeks to scale up and evaluate the ...implementation of shared decision-making (SDM) in interprofessional (IP) home care teams caring for elderly clients or their caregivers facing a decision about staying at home or moving elsewhere.MethodsA stepped wedge cluster randomised trial involving 8 Health and Social Service Centers (HSSCs) will be conducted with IP home care teams. HSSCs are the unit of randomisation. A decision guide will be passively distributed to all of the participating HSSCs at the beginning of the project. The participating HSSCs will then be randomised to 1 of 4 intervention start times, separated by 7-month intervals. The primary outcome is whether or not clients and caregivers assumed an active role in decision-making, assessed with a modified version of the Control Preferences Scale. The intervention, targeted at IP home care teams, consists of a 1.5 hour online tutorial and a 3.5 hour skills building workshop in IP SDM. Clients will be eligible for outcome assessment if they (1) are aged ≥65; (2) are receiving care from the IP home care team of the enrolled HSSCs; (3) have made a decision about whether to stay at home or move to another location during the recruitment periods; (4) are able to read, understand and write French or English; (5) can give informed consent. If clients are not able to provide informed consent, their primary caregiver will become the eligible participant.Ethics and disseminationEthics committee review approval has been obtained from the Multicenter Ethics Committee of CISSS-Laval. Results will be disseminated at conferences, on websites of team members and in peer-reviewed and professional journals intended for policymakers and managers.Trial registration numberNCT02592525, Pre-results.
Joint DNA molecules are natural byproducts of DNA replication and repair. Persistent joint molecules give rise to ultrafine DNA bridges (UFBs) in mitosis, compromising sister chromatid separation. ...The DNA translocase PICH (ERCC6L) has a central role in UFB resolution. A genome-wide loss-of-function screen is performed to identify the genetic context of PICH dependency. In addition to genes involved in DNA condensation, centromere stability, and DNA-damage repair, we identify FIGNL1-interacting regulator of recombination and mitosis (FIRRM), formerly known as C1orf112. We find that FIRRM interacts with and stabilizes the AAA+ ATPase FIGNL1. Inactivation of either FIRRM or FIGNL1 results in UFB formation, prolonged accumulation of RAD51 at nuclear foci, and impaired replication fork dynamics and consequently impairs genome maintenance. Combined, our data suggest that inactivation of FIRRM and FIGNL1 dysregulates RAD51 dynamics at replication forks, resulting in persistent DNA lesions and a dependency on PICH to preserve cell viability.
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•PICH is synthetic lethal with C1orf112/FIRRM•FIRRM forms a complex with the AAA+ ATPase FIGNL1•FIRRM and FIGNL1 control RAD51 dynamics•Persistent RAD51 on chromatin leads to ultrafine bridges and PICH dependency
Persistently connected chromosomes during mitosis threaten chromosome segregation and genomic stability. Joint chromosomes form ultrafine DNA bridges, which are resolved by PICH. Stok et al. discover that C1orf112/FIRRM is synthetic lethal with PICH. FIRMM interacts with FIGNL1 to control RAD51 dynamics and prevent transmission of persistent DNA lesions into mitosis.
BackgroundAdoptive cell transfer (ACT) of neoantigen-reactive tumor-infiltrating lymphocytes (TILs) is an emerging therapeutic modality for solid cancers. A growing body of clinical data in the ...TIL-ACT field supports the potential for the identification, selection, and expansion of tumor-reactive T cells to drive objective response in patients. We believe improvement in the method for identifying neoantigens may further increase the breadth and number of tumor-reactive T cells. Tissue biopsy based neoantigen identification can be limiting due to inter- and intra-tumoral heterogeneity and tissue access. Here, we applied whole exome DNA and RNA sequencing on patient liquid biopsy samples to assess the sensitivity of tumor variant detection and prioritization of neoantigen peptides in comparison with tissue data and to potentially improve target yield.MethodsMatched solid tissue and blood samples were collected from 10 patients (CRC, breast, melanoma, or NSCLC). For solid tissue, whole exome sequencing (WES) and transcriptome libraries were prepared using standard tissue protocols. For blood samples, cell-free DNA (cfDNA) and circulating RNA (cRNA) exome libraries were prepared using Illumina RUO library prep kit reagents. Solid tumor variant calling and neoantigen identification were performed by the Turnstone BFX-4101 platform. Liquid biopsy tumor variants and neoantigens were identified using the DRAGENTM Bio-IT platform and pVACtools suite.ResultsUltra-deep WES (>15,000x) of cfDNA resulted in 100% identification of known small variants at 0.5% variant allele frequency (VAF) in control samples, and >80% sensitivity for 0.2% VAF small variant detection in patient samples. Both analytical pipelines achieved 100% sensitivity on a dataset comprising experimentally determined immunogenic peptides. Concordance of somatic variant calls made between the solid and liquid biopsies ranged from 22%-88% concordance in 4 samples, while 6 samples showed no concordance. Further analysis showed a positive correlation between variant concordance and the percent tumor fraction in the liquid biopsy (from 3.6% to 0% tumor fraction in high to low concordance samples, respectively). For those samples with variant level concordance, up to 40% concordance was observed on neoepitope peptide identification between solid and liquid biopsies. In addition, the liquid biopsy data resulted in up to 29x more peptide calls than the solid tissue, suggesting the blood samples may contain unique tumor fragments not detected in solid tissue biopsy.ConclusionsMinimally invasive liquid biopsy is viable for detection of somatic variants with the potential to broaden selection of tumor-reactive TILs and improve objective response.AcknowledgementsAuthors U. Singh and C. Laing contributed equallyEthics ApprovalThis study was approved by the Advarra IRB; IRB#00000971; IRB#21.299. Informed consent was obtained from all participants.
Improving care transitions for older adults can reduce emergency department (ED) visits, adverse events, and empower community autonomy. We conducted an inductive qualitative content analysis to ...identify themes emerging from comments to better understand ED care transitions.
The LEARNING WISDOM prospective longitudinal observational cohort includes older adults (≥ 65 years) who experienced a care transition after an ED visit from both before and during COVID-19. Their comments on this transition were collected via phone interview and transcribed. We conducted an inductive qualitative content analysis with randomly selected comments until saturation. Themes that arose from comments were coded and organized into frequencies and proportions. We followed the Standards for Reporting Qualitative Research (SRQR).
Comments from 690 patients (339 pre-COVID, 351 during COVID) composed of 351 women (50.9%) and 339 men (49.1%) were analyzed. Patients were satisfied with acute emergency care, and the proportion of patients with positive acute care experiences increased with the COVID-19 pandemic. Negative patient comments were most often related to communication between health providers across the care continuum and the professionalism of personnel in the ED. Comments concerning home care became more neutral with the COVID-19 pandemic.
Patients were satisfied overall with acute care but reported gaps in professionalism and follow-up communication between providers. Comments may have changed in tone from positive to neutral regarding home care over the COVID-19 pandemic due to service slowdowns. Addressing these concerns may improve the quality of care transitions and provide future pandemic mitigation strategies.
Human butyrylcholinesterase (huBChE) has been shown to be an effective antidote against multiple LD50 of organophosphorus compounds. A prerequisite for such use of huBChE is a prolonged circulatory ...half-life. This study was undertaken to produce recombinant huBChE fused to human serum albumin (hSA) and characterize the fusion protein.
Secretion level of the fusion protein produced in vitro in BHK cells was approximately 30 mg/liter. Transgenic mice and goats generated with the fusion constructs expressed in their milk a bioactive protein at concentrations of 0.04-1.1 g/liter. BChE activity gel staining and a size exclusion chromatography (SEC)-HPLC revealed that the fusion protein consisted of predominant dimers and some monomers. The protein was confirmed to have expected molecular mass of approximately 150 kDa by Western blot. The purified fusion protein produced in vitro was injected intravenously into juvenile pigs for pharmacokinetic study. Analysis of a series of blood samples using the Ellman assay revealed a substantial enhancement of the plasma half-life of the fusion protein (approximately 32 h) when compared with a transgenically produced huBChE preparation containing >70% tetramer (approximately 3 h). In vitro nerve agent binding and inhibition experiments indicated that the fusion protein in the milk of transgenic mice had similar inhibition characteristics compared to human plasma BChE against the nerve agents tested.
Both the pharmacokinetic study and the in vitro nerve agent binding and inhibition assay suggested that a fusion protein retaining both properties of huBChE and hSA is produced in vitro and in vivo. The production of the fusion protein in the milk of transgenic goats provided further evidence that sufficient quantities of BChE/hSA can be produced to serve as a cost-effective and reliable source of BChE for prophylaxis and post-exposure treatment.
Obesity is associated with changes in pulmonary function and increased systemic inflammation. We explored the relationships among adiposity, body fat distribution indices, serum inflammatory markers ...and pulmonary function.
This was a post-hoc cross-sectional analysis that included subjects who had previously participated in randomized studies on obesity at our centre. Non-smoking sedentary men (282 subjects, mean age 42) without respiratory diseases were studied. BMI, waist circumference (WC), visceral and subcutaneous adipose tissue (AT), lung residual volume (RV), vital capacity (VC) and expiratory reserve volume (ERV) were measured. Serum leptin, adiponectin, tumor necrosis factor alpha (TNF-α) and high-sensitive C-reactive protein (hs-CRP) levels were measured.
In subjects with metabolic syndrome (n=124), percent predicted ERV and RV were significantly associated with BMI (ERV: r=-0.19, p=0.02, RV: r=-0.28, p=0.0007), WC (r=-0.20, p=0.02, r=-0.26, p=0.002), visceral (r=-0.22, p=0.007, r=-0.25, p=0.002) and subcutaneous AT (r=-0.19, p=0.02, r=-0.28, p=0.0007). Percent predicted VC correlated with visceral (r=-0.20, p=0.02) and subcutaneous AT (r=-0.18, p=0.03). Leptin was strongly correlated with BMI (MS/no-MS: r=0.52, p=0.0005/r=0.62, p < 0.0001), WC (r=0.41, p=0.008/r=0.49, p < 0.0001), visceral (r=0.27,p=0.09/0.43, p < 0.0001) and subcutaneous AT (r=0.46, p=0.003/r=0.66, p < 0.0001), while adiponectin levels were associated in subjects with no-MS with WC (r=-0.20, p=0.01), visceral (r=-0.22, p=0.008), and subcutaneous AT (r=-0.17, p=0.05). When adjusted for anthropometric measures, neither ERV, RV nor VC was significantly correlated with serum leptin, adiponectin, TNF-α, or hs-CRP levels.
These results suggest that the influence of obesity on lung function in healthy subjects is mostly mediated by mechanical factors. Furthermore, not only BMI but also the pattern of fat distribution should be considered when studying associations between adiposity indices and mechanical or inflammatory variables potentially associated with pulmonary function.
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