How the genetic landscape governs a tumor's response to immunotherapy remains poorly understood. To assess the immune-modulatory capabilities of 573 genes associated with altered cytotoxicity in ...human cancers, here we perform CRISPR/Cas9 screens directly in mouse lung cancer models. We recover the known immune evasion factors Stat1 and Serpinb9 and identify the cancer testis antigen Adam2 as an immune modulator, whose expression is induced by Kras
and further elevated by immunotherapy. Using loss- and gain-of-function experiments, we show that ADAM2 functions as an oncogene by restraining interferon and TNF cytokine signaling causing reduced presentation of tumor-associated antigens. ADAM2 also restricts expression of the immune checkpoint inhibitors PDL1, LAG3, TIGIT and TIM3 in the tumor microenvironment, which might explain why ex vivo expanded and adoptively transferred cytotoxic T-cells show enhanced cytotoxic efficacy in ADAM2 overexpressing tumors. Together, direct in vivo CRISPR/Cas9 screens can uncover genetic alterations that control responses to immunotherapies.
Background
The COVID-19 pandemic brought rapid changes to the work and personal lives of clinicians.
Objective
To assess clinician burnout and well-being during the COVID-19 pandemic and guide ...healthcare system improvement efforts.
Design
A survey asking about clinician burnout, well-being, and work experiences.
Participants
Surveys distributed to 8141 clinicians from June to August 2020 in 9 medical groups and 17 hospitals at Sutter Health, a large healthcare system in Northern California.
Main Measures
Burnout was the primary outcome, and other indicators of well-being and work experience were also measured. Descriptive statistics and multivariate logistic regression analyses were performed. All statistical inferences were based on weighted estimates adjusting for response bias.
Key Results
A total of 3176 clinicians (39.0%) responded to the survey. Weighted results showed 29.2% reported burnout, and burnout was more common among women than among men (39.0% vs. 22.7%,
p
<0.01). In multivariate models, being a woman was associated with increased odds of reporting burnout (OR=2.19, 95% CI: 1.51–3.17) and being 55+ years old with lower odds (OR=0.54, 95% CI: 0.34–0.87). More women than men reported that childcare/caregiving was impacting work (32.9% vs. 19.0%,
p
<0.01). Even after controlling for age and gender, clinicians who reported childcare/caregiving responsibilities impacted their work had substantially higher odds of reporting burnout (OR=2.19, 95% CI: 1.54–3.11). Other factors associated with higher burnout included worrying about safety at work, being given additional work tasks, concern about losing one’s job, and working in emergency medicine or radiology. Protective factors included believing one’s concerns will be acted upon and feeling highly valued.
Conclusions
This large survey found the pandemic disproportionally impacted women, younger clinicians, and those whose caregiving responsibilities impacted their work. These results highlight the need for a holistic and targeted strategy for improving clinician well-being that addresses the needs of women, younger clinicians, and those with caregiving responsibilities.
We examined the effects of metformin on diabetes prevention and the subgroups that benefited most over 15 years in the Diabetes Prevention Program (DPP) and its follow-up, the Diabetes Prevention ...Program Outcomes Study (DPPOS).
During the DPP (1996-2001), adults at high risk of developing diabetes were randomly assigned to masked placebo (
= 1,082) or metformin 850 mg twice daily (
= 1,073). Participants originally assigned to metformin continued to receive metformin, unmasked, in the DPPOS (2002-present). Ascertainment of diabetes development was based on fasting or 2-h glucose levels after an oral glucose tolerance test or on HbA
. Reduction in diabetes incidence with metformin was compared with placebo in subgroups by hazard ratio (HR) and rate differences (RDs).
During 15 years of postrandomization follow-up, metformin reduced the incidence (by HR) of diabetes compared to placebo by 17% or 36% based on glucose or HbA
levels, respectively. Metformin's effect on the development of glucose-defined diabetes was greater for women with a history of prior gestational diabetes mellitus (GDM) (HR 0.59, RD -4.57 cases/100 person-years) compared with parous women without GDM (HR 0.94, RD -0.38 cases/100 person-years interaction
= 0.03 for HR,
= 0.01 for RD). Metformin also had greater effects, by HR and RD, at higher baseline fasting glucose levels. With diabetes development based on HbA
, metformin was more effective in subjects with higher baseline HbA
by RD, with metformin RD -1.03 cases/100 person-years with baseline HbA
<6.0% (42 mmol/mol) and -3.88 cases/100 person-years with 6.0-6.4% (
= 0.0001).
Metformin reduces the development of diabetes over 15 years. The subsets that benefitted the most include subjects with higher baseline fasting glucose or HbA
and women with a history of GDM.
The SuperCam instrument suite provides the Mars 2020 rover, Perseverance, with a number of versatile remote-sensing techniques that can be used at long distance as well as within the robotic-arm ...workspace. These include laser-induced breakdown spectroscopy (LIBS), remote time-resolved Raman and luminescence spectroscopies, and visible and infrared (VISIR; separately referred to as VIS and IR) reflectance spectroscopy. A remote micro-imager (RMI) provides high-resolution color context imaging, and a microphone can be used as a stand-alone tool for environmental studies or to determine physical properties of rocks and soils from shock waves of laser-produced plasmas. SuperCam is built in three parts: The mast unit (MU), consisting of the laser, telescope, RMI, IR spectrometer, and associated electronics, is described in a companion paper. The on-board calibration targets are described in another companion paper. Here we describe SuperCam’s body unit (BU) and testing of the integrated instrument.
The BU, mounted inside the rover body, receives light from the MU via a 5.8 m optical fiber. The light is split into three wavelength bands by a demultiplexer, and is routed via fiber bundles to three optical spectrometers, two of which (UV and violet; 245–340 and 385–465 nm) are crossed Czerny-Turner reflection spectrometers, nearly identical to their counterparts on ChemCam. The third is a high-efficiency transmission spectrometer containing an optical intensifier capable of gating exposures to 100 ns or longer, with variable delay times relative to the laser pulse. This spectrometer covers 535–853 nm (
105
–
7070
cm
−
1
Raman shift relative to the 532 nm green laser beam) with
12
cm
−
1
full-width at half-maximum peak resolution in the Raman fingerprint region. The BU electronics boards interface with the rover and control the instrument, returning data to the rover. Thermal systems maintain a warm temperature during cruise to Mars to avoid contamination on the optics, and cool the detectors during operations on Mars.
Results obtained with the integrated instrument demonstrate its capabilities for LIBS, for which a library of 332 standards was developed. Examples of Raman and VISIR spectroscopy are shown, demonstrating clear mineral identification with both techniques. Luminescence spectra demonstrate the utility of having both spectral and temporal dimensions. Finally, RMI and microphone tests on the rover demonstrate the capabilities of these subsystems as well.
Stable primary functionalization of metal surfaces plays a significant role in reliable secondary attachment of complex functional molecules used for the interfacing of metal objects and ...nanomaterials with biological systems. In principle, this can be achieved through chemical reactions either in the vapor or liquid phase. In this work, we compared these two methods for oxidized silicon surfaces and thoroughly characterized the functionalization steps by tagging and fluorescence imaging. We demonstrate that the vapor‐phase functionalization only provided transient surface modification that was lost on extensive washing. For stable surface modification, a liquid‐phase method was developed. In this method, silicon wafers were decorated with azides, either by silanization with (3‐azidopropyl)triethoxysilane or by conversion of the amine groups of an aminopropylated surface by means of the azido‐transfer reaction. Subsequently, D‐amino acid adhesion peptides could be immobilized on the surface by use of CuI‐catalyzed click chemistry. This enabled the study of cell adhesion to the metal surface. In contrast to unmodified surfaces, the peptide‐modified surfaces were able to maintain cell adhesion during significant flow velocities in a microflow reactor.
Stuck tight: The fully optimized coating of silicon surfaces with a monolayer of (3‐azidopropyl)siloxide allowed attachment of secondary complex molecules. D‐Amino acid adhesion peptides k‐l‐h‐r‐l‐r‐a and k‐l‐y‐r‐v‐r‐a were immobilized through CuAAC, and their interaction with cells was investigated by flow shear. Stable adhesion of HEK293 cells was achieved even at high flow rates.
Trichoderma reesei is the main industrial source of cellulases and hemicellulases used to depolymerize biomass to simple sugars that are converted to chemical intermediates and biofuels, such as ...ethanol. We assembled 89 scaffolds (sets of ordered and oriented contigs) to generate 34 Mbp of nearly contiguous T. reesei genome sequence comprising 9,129 predicted gene models. Unexpectedly, considering the industrial utility and effectiveness of the carbohydrate-active enzymes of T. reesei, its genome encodes fewer cellulases and hemicellulases than any other sequenced fungus able to hydrolyze plant cell wall polysaccharides. Many T. reesei genes encoding carbohydrate-active enzymes are distributed nonrandomly in clusters that lie between regions of synteny with other Sordariomycetes. Numerous genes encoding biosynthetic pathways for secondary metabolites may promote survival of T. reesei in its competitive soil habitat, but genome analysis provided little mechanistic insight into its extraordinary capacity for protein secretion. Our analysis, coupled with the genome sequence data, provides a roadmap for constructing enhanced T. reesei strains for industrial applications such as biofuel production.
Functional immune responses are increasingly important for clinical studies, providing in depth biomarker information to assess immunotherapy or vaccination. Incorporating functional immune assays ...into routine clinical practice has remained limited due to challenges in standardizing sample preparation. We recently described the use of a whole blood syringe-based system, TruCulture®, which permits point-of-care standardized immune stimulation. Here, we report on a multi-center clinical study in seven FOCIS Centers of Excellence to directly compare TruCulture to conventional PBMC methods. Whole blood and PBMCs from healthy donors were exposed to LPS, anti-CD3 anti-CD28 antibodies, or media alone. 55 protein analytes were analyzed centrally by Luminex multi-analyte profiling in a CLIA-certified laboratory. TruCulture responses showed greater reproducibility and improved the statistical power for monitoring differential immune response activation. The use of TruCulture addresses a major unmet need through a robust and flexible method for immunomonitoring that can be reproducibly applied in multi-center clinical studies.
A multi-center study revealed greater reproducibility from whole blood stimulation systems as compared to PBMC stimulation for studying induced immune responses.
•FOCIS Centers of Excellence clinical study compared standardized whole blood TruCulture and PBMC stimulations•TruCulture showed greater reproducibility across centers and improved statistical power for studying immune responses•Significantly lower levels of non-specific cytokines were observed in whole blood stimulation as compared to PBMC cultures•Significantly higher IL-23 and CXCL5 for LPS, and lower VEGF for CD3/CD28 stimulation in TruCulture as compared to PBMC
Research suggests striking disparities in alcohol use, problems, and treatment across racial/ethnic groups in the United States. However, research on alcohol-related disparities affecting ...racial/ethnic minorities remains in its developmental stages. The current article aims to support future research in this growing field by highlighting some of the most important findings, questions, and approaches, focusing on psychosocial research.
This article advances seven research needs (i.e., questions and topics meriting attention) that we believe are of crucial importance to the field. We draw on the existing literature to illuminate under-explored areas that are highly relevant to health intervention and that complement the field's existing focus.
Identified research needs include research that (a) better describes disparities in alcohol-related health conditions and their drivers, (b) identifies appropriate screening and brief intervention methods for racial/ethnic minorities, (c) investigates disparities in access to and use of alcohol treatment and support services, (d) examines the comparative efficacy of existing alcohol interventions and develops tailored interventions, (e) explores the impacts of specific alcohol policies across and within racial/ethnic groups, and (f) describes the full spectrum of alcohol-related harms and how and why these may vary across racial/ethnic groups. We also call for (g) continuing research to monitor disparities over time.
This article points to specific strategies for describing, explaining, intervening on, and monitoring some of the most substantial alcohol-related disparities. Conclusions outline methods and processes that may be advantageous in addressing these priorities, including the use of longitudinal designs; consideration of life course changes; attention to nontraditional intervention settings; and inclusion of disadvantaged populations in all aspects of research.
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