Search

Autoimmune diseases are characterized by a breakdown of immune tolerance partly due to environmental factors. The short-chain fatty acid acetate, derived mostly from gut microbial fermentation of dietary fiber, promotes antiinflammatory Tregs and protects mice from type 1 diabetes, colitis, and allergies. Here, we show that the effects of acetate extend to another important immune subset involved in tolerance, the IL-10–producing regulatory B cells (B10 cells). Acetate directly promoted B10 cell differentiation from mouse B1a cells both in vivo and in vitro. These effects were linked to metabolic changes through the increased production of acetyl-coenzyme A, which fueled the TCA cycle and promoted posttranslational lysine acetylation. Acetate also promoted B10 cells from human blood cells through similar mechanisms. Finally, we identified that dietary fiber supplementation in healthy individuals was associated with increased blood-derived B10 cells. Direct delivery of acetate or indirect delivery via diets or bacteria that produce acetate might be a promising approach to restore B10 cells in noncommunicable diseases.

The chief goal of the Blood Profiling Atlas in Cancer (BloodPAC) consortium is to promote collaborative efforts that support the development and implementation of liquid biopsy tests. Here, we report the results of a pilot study conducted by three BloodPAC members that aimed to demonstrate a multisite liquid biopsy testing framework using longitudinal blood specimens from 38 patients with metastatic breast cancer. Three laboratories receiving identical samples from two clinical sites each applied a different targeted sequencing platform to analyze mutations in cell-free DNA (cfDNA). The resulting mutational profiles reflected common breast cancer alterations, including clinically actionable mutations for 40% of hormone- receptor-positive patients. In 12 genes with shared target regions across sequencing panels, perfect inter-assay concordance was also observed for mutations detected above the lowest common assay limit of detection. Whole-genome copy number profiling of cfDNA and circulating tumor cells (CTCs) further revealed marked heterogeneity in copy number alterations and cfDNA tumor fractions across patients. Additionally, comparison of tumor fraction and CTC abundance demonstrated the complementary nature of cfDNA and CTC analyses. Overall, the framework described in this study may serve as a resource for future trials aiming to identify multimodal liquid biopsy biomarkers to guide clinical care.

We report the observation of a nonlinear terahertz response of split-ring resonator arrays made of high-temperature superconducting films. Intensity-dependent transmission measurements indicate that the resonance strength decreases dramatically (i.e. transient bleaching) and the resonance frequency shifts as the intensity is increased. Pump–probe measurements confirm this behaviour and reveal dynamics on the few-picosecond timescale.

We report the observation of a nonlinear terahertz response of split-ring resonator arrays made of high-temperature superconducting films. Intensity-dependent transmission measurements indicate that the resonance strength decreases dramatically (i.e. transient bleaching) and the resonance frequency shifts as the intensity is increased. Pump–probe measurements confirm this behaviour and reveal dynamics on the few-picosecond timescale. [ABSTRACT FROM AUTHOR]

Rotational orientation/alignment dynamics of CO2scattered from a perfluorinated polyether (PFPE) liquid surface has been investigated via direct absorption experimental studies and theoretical molecular dynamics (MD) simulations. Experimentally, polarization modulation of a single mode diode laser is combined with lock-in detection to measure circular/linear IR polarizance due to CO2scattering from the surface at θinc= 60° and Einc= 10.6(8) kcal/mol and probed over a series of final scattering angles. The differential absorption intensities are related through Fano−Macek theory to the three lowest multipole moments (A0, A2+, and O1−) which describe collisionally induced orientation and alignment at the liquid surface. The total scattering population reflects both trapping-desorption (TD) and impulsive scattering (IS) components, with a strong positiveanisotropy in the MJdistribution that indicates preferential CO2scattering from the surface with a forward(i.e., “topspin”) sense of end-over-end tumbling. Theoretical trajectory simulations provide 3D CO2flux and Jstate distributions scattering from fluorinated self-assembled monolayers (F-SAMs) and are compared with experimental results as a function of final rotational state. Specifically, trends in the theoretical orientation/alignment moments are in remarkable agreement over the full range of J states but with values consistently overpredicted by nearly 2-fold, which may reflect a higher level of local ordering for F-SAMS vs a gas−PFPE liquid interface. [ABSTRACT FROM AUTHOR]

Molecular beam scattering dynamics at the gas−liquid interface are investigated for CO 2( Einc= 10.6(8) kcal/mol) impinging on liquid perfluoropolyether (PFPE), with quantum state ( v, J) populations measured as a function of incident (θ inc) and final (θ scat) scattering angles. The internal state distributions are well-characterized for both normal and grazing incident angles by a two-component Boltzmann model for trapping desorption (TD) and impulsive scattering (IS) at rotational temperatures Trot(TD/IS), where the fractional TD probability for CO 2on the perfluorinated surface is denoted by TD and IS densities (ρ) as α = ρ TD/(ρ TD+ ρ IS). On the basis of an assumed cos(θ scat) scattering behavior for the TD flux component, the angular dependence of the IS flux at normal incidence (θ inc= 0°) is surprisingly well-modeled by a simple cos n(θ scat) distribution with n= 1.0 ± 0.2, while glancing incident angles (θ inc= 30°, 45°, and 60°) result in lobular angular IS distributions scattered preferentially in the forward direction. This trend is also corroborated in the TD fraction α, which decreases rapidly under non-normal incident conditions as a function of backward versus forward scattering direction. Furthermore, the extent of rotational excitation in the IS channel increases dramatically with increasing angle of incidence, consistent with an increasing rotational torque due to surface roughness at the gas−liquid interface. [ABSTRACT FROM AUTHOR]

Energy transfer dynamics at the gas−liquid interface are investigated as a function of surface temperature both by experimental studies of CO2perfluorinated polyether (PFPE) and by molecular dynamics simulations of CO2fluorinated self-assembled monolayers (F-SAMs). Using a normal incident molecular beam, the experimental studies probe scattered CO2internal-state and translational distributions with high resolution infrared spectroscopy. At low incident energies Einc1.6(1) kcal/mol, CO2J-state populations and transverse Doppler velocity distributions are characteristic of the surface temperature (TrotTtransTS) over the range from 232 to 323 K. In contrast, the rotational and translational distributions at high incident energies Einc10.6(8) kcal/mol show evidence for both trapping-desorption (TD) and impulsive scattering (IS) events. Specifically, the populations are surprisingly well-characterized by a sum of Boltzmann distributions where the two components include one (TD) that equilibrates with the surface (TTDTS) and a second (IS) that is much hotter than the surface temperature (TISTS). Support for the superthermal, yet Boltzmann, nature of the IS channel is provided by molecular dynamics (MD) simulations of CO2F-SAMs Einc10.6 kcal/mol, which reveal two-temperature distributions, sticking probabilities, and angular distributions in near quantitative agreement with the experimental PFPE results. Finally, experiments as a function of surface temperature reveal an increase in both sticking probability and rotational/translational temperature of the IS component. Such a trend is consistent with increased surface roughness at higher surface temperature, which increases the overall probability of trapping, yet preferentially leads to impulsive scattering of more highly internally excited CO2from the surface. [ABSTRACT FROM AUTHOR]

Energy transfer dynamics at the gas−liquid interface have been probed with a supersonic molecular beam of CO2and a clean perfluorinated-liquid surface in vacuum. High-resolution infrared spectroscopy measures both the rovibrational state populations and the translational distributions for the scattered CO2flux. The present study investigates collision dynamics as a function of incident angle (inc0°, 30°, 45°, and 60°), where column-integrated quantum state populations are detected along the specular-scattering direction (i.e., scatinc). Internal state rovibrational and Doppler translational distributions in the scattered CO2yield clear evidence for nonstatisticalbehavior, providing quantum-state-resolved support for microscopic branching of the gas−liquid collision dynamics into multiple channels. Specifically, the data are remarkably well described by a two-temperature model, which can be associated with both a trapping desorption (TD) component emerging at the surface temperature (TrotTS) and an impulsive scattering (IS) component appearing at hyperthermal energies (Trot> TS). The branching ratio between the TD and IS channels is found to depend strongly on inc, with the IS component growing dramatically with increasingly steeper angle of incidence. [ABSTRACT FROM AUTHOR]

Quantum-state-resolved dynamics at the gas−liquid interface are probed by colliding supersonically cooled molecular beams of CO2with freshly formed liquid surfaces in a vacuum. Translational, rotational, and vibrational state distributions of both incident and scattered fluxes are measured by high-resolution direct infrared absorption spectroscopy and laser dopplerimetry in the 0000 and 0110 rovibrational manifolds of CO2in the asymmetric stretch manifold. The present studies investigate the role of incident molecular beam energy (Einc 1.6(1), 4.7(2), 7.7(2), and 10.6(8) kcal/mol) on these distributions for a series of perfluorinated, hydrocarbon, and hydrogen-bonded liquids. Boltzmann analysis of the internal quantum-state populations provide evidence for nonthermal scattering dynamics, as confirmed by Dopplerimetry on the absorption profiles. The data provide quantum-state-resolved support for a dual channel picture of the scattering process, consisting of either prompt impulsive scattering (IS) or longer duration trapping−desorption (TD) events, with the fraction observed in each channel dependent on incident kinetic energy and the physical properties of the liquid surface. The clear evidence that internal CO2rotational populations arising from the IS channel can be adequately described by a Boltzmann temperature (albeit with EIS> RTS) is consistent with previous gas−solid scattering studies and suggests that even nominally “prompt” IS events reflect both single(i.e. direct) and multipleimpulsive interactions with the liquid interface. [ABSTRACT FROM AUTHOR]

Harvesting of clams(Tapes philippinarum) has important socio-economic and environmental implications for the Venice lagoon area, Italy. Clam harvesting disrupts the structure of benthic communities but the effects upon sediment stability and surface structure remain unclear. The effect of clam fishing on the sediment properties of the lagoon bed was investigated at two different sites, a heavily fished site (San Angelo) and an infrequently fished site (San Giaccomo). Both sites were assessed for immediate impacts of fishing, using indicators of biogenic sediment stabilisation. Samples were taken at three points along three 100 m linear transects at each site prior to and post fishing. Paired samples were also taken parallel to each transect at a distance of 5m, to allow for temporal variation. Sediment stability, measured with a cohesive strength meter (CSM), was significantly higher at the less impacted site (F1,34 = 6.23, p < 0.018), was correlated with indicators of biogenic sediment stabilisation and decreased by approximately 50% following the trawling event. Concomitant decreases in chlorophyll a (chl a), colloidal-S carbohydrate and dry bulk density were observed on the transect after fishing but not adjacent to the fishing path. At the heavily impacted site, clam fishing by trawling had, in general, no significant effect on the biological and physical properties (although chl a did decrease significantly after fishing). The lack of a significant impact from fishing at the impacted site was attributed to the higher frequency of fishing occurring in this area. Hence, frequent fishing of the lagoon prevents establishment of biotic communities, preventing biostabilisation and thus reduces the stability of the surface sediment. Keywords: clam harvesting, erosion threshold, microphytobenthos, sediment, stability

Tumor volume has been suggested as an important prognostic factor of prostatic adenocarcinoma (PAC) treated with radical prostatectomy (RP). The calculation of tumor volume is complicated by the difficulty in appreciation of tumor nodules at gross examination, multifocality, and variation in the shape of tumor nodules. We propose a simple technique for the calculation of tumor volume. One hundred consecutive specimens of RP were studied with special attention to the shape of tumor nodules. Most small PAC, transitional zone (TZ) PAC, peripheral zone (PZ) PAC without associated benign prostatic hyperplasia (BPH), and PZPAC with Gleason's score (GS) > 3 + 4 had an ovoid shape. Most large sized nodules of PZPAC with GS < 4 + 3 tended to mold according to the boundaries of the TZ that were themselves often compressed by hyperplastic nodules. Therefore, these large tumor nodules were crescentically shaped and had tapering pole(s). We deduced from that tendency that the ratio of height of the tumor nodule = D1 × the height/greatest horizontal diameter of the prostate (D1 = the greatest diameters of the largest section of tumor nodule). Using the mathematical formula for volume of an ellipsoid structure, we propose the following formula to calculate the volume of each tumor nodule = 0.8 × K × D12 × D2 (D2 = greatest diameter orthogonal to D1, and K = coefficient for correction of tumor volume due to the compression of hyperplastic nodules). K is empirically estimated as 2/3 for PZPAC in mid prostate and 1/2 for tumor nodules at the apex and base. The total tumor volume is the sum of all tumor nodule volumes. By measuring the two greatest orthogonal diameters, D1 and D2, of the largest horizontal section of a tumor nodule, we were able to calculate the corresponding volume and consequently the total tumor volume of the prostate. Analysis of the calculated total tumor volume showed a good correlation with the current technique of measurement on each section of the prostate, particularly for tumors ranging from 1.5 to 3.0 cm3. [ABSTRACT FROM AUTHOR]

Caloric restriction (CR) can extend the organism life- and health-span by improving glucose homeostasis. How CR affects the structure-function of pancreatic beta cells remains unknown. We used single nucleus transcriptomics to show that CR increases the expression of genes for beta cell identity, protein processing, and organelle homeostasis. Gene regulatory network analysis reveal that CR activates transcription factors important for beta cell identity and homeostasis, while imaging metabolomics demonstrates that beta cells upon CR are more energetically competent. In fact, high-resolution microscopy show that CR reduces beta cell mitophagy to increase mitochondria mass and the potential for ATP generation. However, CR beta cells have impaired adaptive proliferation in response to high fat diet feeding. Finally, we show that long-term CR delays the onset of beta cell aging hallmarks and promotes cell longevity by reducing beta cell turnover. Therefore, CR could be a feasible approach to preserve compromised beta cell structure-function during aging and diabetes., (© 2024. The Author(s).)

Background: Immune checkpoint inhibitors (ICIs) are the guideline endorsed first choice for patients with deficient mismatch repair or microsatellite instability high (dMMR/MSI-H) mCRC, however a significant proportion experience primary or secondary resistance. BRAF V600E mutated (BRAFm) and dMMR/MSI-H mCRC can be treated with BRAF + EGFR inhibitors but specific data on the efficacy after progression to ICIs are missing., Methods: We collected consecutive patients with BRAFm dMMR/MSI-H mCRC treated from 2017 to 2024 with a combination of BRAFi+EGFRi+/-MEKi, after disease progression on ICIs. A control cohort of BRAFm pMMR/MSS mCRC patients treated with encorafenib+cetuximab+/-binimetinib from 2nd line was used., Results: dMMR/MSI-H (n = 50) BRAFm mCRC patients were more often > 70-year-old, with right-sided primary tumors, without liver but more lymphnode metastases than pMMR/MSS (n = 170). They were treated more frequently beyond 2nd line and 45 % were primary progressors to ICIs. Lower ORR (18 % versus 32 %, p = 0.09) and DCR (60 % versus 73 %, p = 0.11) was seen without reaching significance in dMMR/MSI-H as compared to pMMR/MSS patients. After a median follow-up of 14.04 months, no differences in PFS (median 5.13 versus 4.50 months, HR 0.83, 95 %CI: 0.57-1.20, p = 0.31) and OS (median 10.75 versus 9.11 months, HR 0.89, 95 %CI: 0.59-1.32, p = 0.55) were observed., Conclusions: Our results show that BRAFm dMMR/MSI-H mCRC patients benefit from BRAFi+EGFRi+/-MEKi after progression under ICIs. Despite lower ORR and DCR, the outcome is not different from that observed in pMMR/MSS BRAFm CRC and is in line with the results of the BEACON registration trial., Competing Interests: Declaration of Competing Interest DT reports personal fees from AstraZeneca, Sanofi, Amgen, BMS, MSD, Roche, Servier, and Pierre Fabre. DM participated in advisory board for Amgen, Servier, Merck, MSD, Takeda, G1, Onkowissen, Pierre Fabre, AstraZeneca, Regeneron; received honoraria as invited speaker for Medison, COR2ED, JE, 21up, Seagen, Takeda, Taiho, Amgen, Servier, Merck, Onkowissen, MSD, AstraZeneca, Pierre Fabre, GSK; reports institutional non-financial interests for Amgen and Servier. GP reports grant from Merck Serono, honoraria from Servier and Sanofi. EE has received personal honoraria from Amgen, Bayer, BMS, Boehringer Ingelheim, Cure Teq AG, Hoffman La – Roche, Janssen, Lilly, Medscape, Merck Serono, MSD, Novartis, Organon, Pfizer, Pierre Fabre, Repare Therapeutics Inc., RIN Institute Inc., Sanofi, Seagen International, GmbH, Servier, and Takeda. CC served as expert testimony for Amgen; received honoraria as invited speaker for Bayer, Merck Serono, Servier; participated in advisory board for MSD, Nordic Pharma, Pierre- Fabre, Roche, Takeda; reports institutional financial interest for Bayer as Coordinating PI, Hutchinson as Local PI, Merck as Coordinating PI, Roche as Coordinating PI, Seagen as Local PI, Servier as Coordinating PI. FP reported receiving institutional research grants from BMS, Incyte, Agenus, Amgen, Lilly and AstraZeneca, and personal fees from BMS, MSD, Amgen, Merck-Serono, Pierre-Fabre, Servier, Bayer, Takeda, Astellas, Johnson&Johnson, Rottapharm, Ipsen, AstraZeneca, GSK, Daiichi-Sankyo, Seagen/Pfizer, Beigene. SL reports research funding to institution from Amgen, Astellas, AstraZeneca, Bayer, Bristol-Myers Squibb, Daichii Sankyo, Hutchinson, Incyte, Merck Serono, Mirati, MSD, Pfizer, Roche, Servier; personal honoraria as invited speaker from Amgen, Bristol- Myers Squibb, Incyte, GSK, Lilly, Merck Serono, MSD, Pierre- Fabre, Roche, Servier; participation in advisory board for Amgen, Astellas, Astra Zeneca, Bayer, Bristol-Myers Squibb, Daiichi-Sankyo, GSK, Incyte, Lilly, Merck Serono, MSD, Servier, Takeda. CG has participated in consulting and/or advisory boards for Servier, Sanofi, Merck, Pierre Fabre, MSD and has received support for travel to meetings from Pierre Fabre and Servier. JT received honoraria as invited speaker for Amgen, Astellas, BMS, Merk, MSD, Novartis; participated in advisory board for Amgen, BMS, Merck, MSD, Novartis, Pfizer, Pierre Fabre, Rottapharm, Sanofi, Servier, Takeda and as expert testimony for Takeda; participated in steering committee of clinical trial for Novartis. All the other authors declare no competing interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objectives: To measure community attitudes to emergency care and treatment plans (ECTPs)., Design: Population survey., Setting: Great Britain., Participants: As part of the British Social Attitudes Survey, sent to randomly selected addresses in Great Britain, 1135 adults completed a module on ECTPs. The sample was nationally representative in terms of age and location, 619 (55%) were female and 1005 (89%) were of white origin., Outcome Measures: People's attitudes having an ECTP for themselves now, and in the future; how comfortable they might be having a discussion about an ECTP and how they thought such a plan might impact on their future care., Results: Predominantly, respondents were in favour of people being able to have an ECTP, with 908/1135 (80%) being at least somewhat in favour. People in good health were less likely than those with activity-limiting chronic disease to want a plan at present (52% vs 64%, OR 1.78 (95% CI 1.30 to 2.45) p<0.001). Developing a long-term condition or becoming disabled would lead 42% (467/1112) and 43% (481/1112) of individuals, respectively, to want an ECTP. More, 634/1112 (57%) would want an ECTP if they developed a life-threatening condition. Predominantly, 938/1135 (83%) respondents agreed that an ECTP would help avoid their family needing to make difficult decisions on their behalf, and 939/1135 (83%) that it would ensure doctors and nurses knew their wishes. Nevertheless, a small majority-628/1135 (55%)-agreed that there was a serious risk of the plan being out of date when needed. A substantial minority-330/1135 (29%)-agreed that an ECTP might result in them not receiving life-saving treatment., Conclusions: There is general support for the use of ECTPs by people of all ages. Nevertheless, many respondents felt these might be out of date when needed and prevent people receiving life-saving treatment., Competing Interests: Competing interests: MU is chief investigator or co-investigator on multiple previous and current research grants from the UK National Institute for Health Research (NIHR), and is a co-investigator on grants funded by the Australian NHMRC and Norwegian MRC. He was an NIHR Senior Investigator until March 2021. He is a director and shareholder of Clinvivo that provides electronic data collection for health services research. He receives some salary support from University Hospitals Coventry and Warwickshire. He is a co-investigator on two current and one completed NIHR-funded studies that have, or have had, additional support from Stryker. AN is co-investigator on multiple research grants funded by the UK NIHR. CB is Chief Executive of Health Watch Warwickshire. JH is co-investigator on multiple grants funded by the UK NIHR. FG is co-investigator on multiple grants funded by the UK NIHR. CH has no conflict of interest to declare. GP is chief investigator or co-investigator on multiple previous and current research grants from the UK NIHR. He is Trustee of the Resuscitation Council UK, Director of the European Resuscitation Council, and co-chair of the International Committee on Resuscitation. SR is co-investigator on multiple grants funded by the UK NIHR. A-MS is chief investigator or co-investigator on multiple previous and current research grants from the UK NIHR. She is a member of the Resuscitation Council UK ReSPECT wider stakeholder group (from March 2024)., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.)

Background: The combination of encorafenib with cetuximab has become the standard of care in patients with BRAF V600E-mutated metastatic colorectal cancer (mCRC) after a prior systemic therapy. This study aims to describe the efficacy and safety of encorafenib/cetuximab +/- binimetinib in patients with BRAF V600E-mutated mCRC in a real-world setting., Patients and Methods: This retrospective study included patients with BRAF V600E-mutated mCRC who received this combination from January 2020 to June 2022 in 30 centers., Results: A total of 201 patients were included, with 55% of women, a median age of 62 years, and an Eastern Cooperative Oncology Group performance status (ECOG-PS) >1 in 20% of cases. The main tumor characteristics were 60% of right-sided primary tumor, 11% of microsatellite instability/mismatch repair deficient phenotype, and liver and peritoneum being the two main metastatic sites (57% and 51%). Encorafenib/cetuximab +/- binimetinib was prescribed in the first, second, third, and beyond third line in 4%, 56%, 29%, and 11%, respectively, of cases, with the encorafenib/cetuximab/binimetinib combination for 21 patients (10%). With encorafenib/cetuximab treatment, 21% of patients experienced grade ≥3 adverse events (AEs), with each type of grade ≥3 AE observed in <5% of patients. The objective response rate was 32.2% and the disease control rate (DCR) was 71.2%. The median progression-free survival (PFS) was 4.5 months [95% confidence interval (CI) 3.9-5.4 months] and the median overall survival (OS) was 9.2 months (95% CI 7.8-10.8 months). In multivariable analysis, factors associated with a shorter PFS were synchronous metastases [hazard ratio (HR) 1.66, P = 0.04] and ECOG-PS >1 (HR 1.88, P = 0.007), and those associated with a shorter OS were the same factors (HR 1.71, P = 0.03 and HR 2.36, P < 0.001, respectively) in addition to treatment beyond the second line (HR 1.74, P = 0.003) and high carcinoembryonic antigen level (HR 1.72, P = 0.003)., Conclusion: This real-world study showed that in patients with BRAF V600E-mutated mCRC treated with encorafenib/cetuximab +/- binimetinib, efficacy and safety data confirm those reported in the BEACON registration trial. The main poor prognostic factors for this treatment are synchronous metastases and ECOG-PS >1., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Management of ampullary tumors (AT) is challenging because of a low level of scientific evidence. This document is a summary of the French intergroup guidelines regarding the management of AT, either adenoma (AA) or carcinoma (AC), published in July 2023, available on the website of the French Society of Gastroenterology (SNFGE) (www.tncd.org)., Methods: A collaborative work was conducted under the auspices of French medical, endoscopic, oncological and surgical societies involved in the management of AT. Recommendations are based on recent literature review and expert opinions and graded in three categories (A, B, C), according to quality of evidence., Results: Accurate diagnosis of AT requires at least duodenoscopy and EUS. All patients should be discussed in multidisciplinary tumor board before treatment. Surveillance may only be proposed for small AA in familial adenomatous polyposis. For AA, endoscopic papillectomy is the preferred option only if R0 resection can be achieved. When not possible, surgical papillectomy should be considered. For AC beyond pT1a N0, pancreaticoduodenectomy is the procedure of choice. Adjuvant monochemotherapy (gemcitabine, 5FU) may be proposed. For aggressive tumors (pT3/T4, pN+, R1, poorly differentiated AC, pancreatobiliary differentiation) with high risk of recurrence, 6 months polychemotherapy (CAPOX/FOLFOX for the intestinal subtype and mFOLFIRINOX for the pancreatobiliary or the mixed subtype) may be a valid alternative. Clinical and radiological follow up is recommended for 5 years., Conclusions: These guidelines help to homogenize and highlight unmet needs in the management of AA and AC. Each individual case should be discussed by a multidisciplinary team., Competing Interests: Conflict of interest VH: AAA, Amgen, Esteve, Ipsen, Deciphera, Merck, Pierre Fabre, Servier AT: personal fees from Servier, Viatris, Incyte Bioscience, BMS, Merck and grants from AstraZeneca and MSD outside the submitted work MC: Medtronic, Boston Scientific, Cook Medical, AMBU CN: Honoraria / consulting: Amgen, AstraZeneca, Baxter, Bristol-Myers Squibb, Fresenius Kabi, Incyte Biosciences, Merck, MSD, Mundipharma, Novartis, Nutricia, OSE Immunotherapeutics, Pierre Fabre, Roche, Sanofi, Servier, Viatris. Research funding / clinical trials: AstraZeneca, Bristol-Myers Squibb, Fresenius Kabi, Nutricia, OSE Immunotherapeutics, Roche, Servier, Viatris. ALB declares consulting for Astra Zeneca , Servier, Ipsen and accomodations or congress registration support for Merck and Ipsen DS: Honoraria / consulting: Amgen, Bayer, Deciphera Pharmaceutical, Ipsen, Pierre fabre, Roche, Servier, Viatris, Travel accomodations: AstraZeneca, Ipsen, Novartis, Pfizer, Pierre Fabre, Roche, Sandoz, Servier GP: honoraria from Servier, Roche and Sanofi OB: honoraria from Amgen, Apmonia Therapeutics, Bayer, Deciphera, Merck KGaA, MSD, Pierre Fabre, Servier. SG: Mylan, IPSEN All other authors declare no conflict of interest related to this article., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Aim: To determine the effect of CPR delivery surface (e.g. firm mattress, floor, backboard) on patient outcomes and CPR delivery., Methods: We searched MEDLINE, Embase, Web of Science and the Cochrane Central Register of Controlled Trials for studies published since 2019 that evaluated the effect of CPR delivery surface in adults and children on patient outcomes and CPR depth (PROSPERO CRD42023467583). We included manikin studies due to a lack of human studies. We identified pre-2019 studies from the 2020 ILCOR evaluation of this topic. Two reviewers independently screened titles/abstracts and full-text papers, extracted data and assessed risk of bias. Evidence certainty for each outcome was evaluated using GRADE methodology. Where appropriate, we pooled data in a meta-analysis, using a random-effects model., Results: Database searches identified 489 citations. We included six studies published since 2019. We analysed these studies together with the eleven studies included in the previous ILCOR review. All included studies were manikin randomised controlled trials. Certainty of evidence was low. Interventions including placing the patient on the floor or the use of backboard had minimal impact on achieving greater compression depth. Meta-analyses of floor versus firm hospital mattress or firm home mattress found a mean difference of 5.36 mm (95% CI -1.59 to 12.32) and 2.11 mm (95% CI -3.23 to 7.45) respectively., Conclusion: The use of a backboard led to a small 2 mm increase in chest compression depth in meta-analysis of multiple mannikin trials. Use of a firm mattress or transitioning to the floor did not affect chest compression depth., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: ‘GDP is an Editor of Resuscitation and Editor-in-Chief of Resuscitation Plus. JB is an Editor of Resuscitation Plus and an Editorial Board Member of Resuscitation. GDP is supported by the National Institute for Health Research (NIHR) Applied Research Collaboration (ARC) West Midlands. The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care.’., (© 2024 The Author(s).)

Upstream open reading frames (uORFs) are cis-acting elements that can dynamically regulate the translation of downstream ORFs by suppressing downstream translation under basal conditions and, in some cases, increasing downstream translation under stress conditions. Computational and empirical methods have identified uORFs in the 5'-UTRs of approximately half of all mouse and human transcripts, making uORFs one of the largest regulatory elements known. Because the prevailing dogma was that eukaryotic mRNAs produce a single functional protein, the peptides and small proteins, or microproteins, encoded by uORFs were rarely studied. We hypothesized that a uORF in the SLC35A4 mRNA is producing a functional microprotein (SLC35A4-MP) because of its conserved amino acid sequence. Through a series of biochemical and cellular experiments, we find that the 103-amino acid SLC35A4-MP is a single-pass transmembrane inner mitochondrial membrane (IMM) microprotein. The IMM contains the protein machinery crucial for cellular respiration and ATP generation, and loss of function studies with SLC35A4-MP significantly diminish maximal cellular respiration, indicating a vital role for this microprotein in cellular metabolism. The findings add SLC35A4-MP to the growing list of functional microproteins and, more generally, indicate that uORFs that encode conserved microproteins are an untapped reservoir of functional microproteins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: One randomized phase III trial comparing chemotherapy (CT) with immune checkpoint inhibitors (ICI) has demonstrated significant efficacy of ICI in deficient DNA mismatch repair system/microsatellite instability-high (dMMR/MSI-H) metastatic colorectal cancer. However, few studies have compared ICI with CT in other advanced dMMR/MSI-H digestive tumors., Methods: In this multicenter study, we included patients with advanced dMMR/MSI-H non-colorectal digestive tumors treated with chemotherapy and/or ICIs. Patients were divided retrospectively into two groups, a CT group and an immunotherapy (IO) group. The primary endpoint was progression-free survival (PFS). A propensity score approach using the inverse probability of treatment weighting (IPTW) method was applied to deal with potential differences between the two groups., Results: 133 patients (45.1/27.1/27.8% with gastric/small bowel/other carcinomas) were included. The majority of patients received ICI in 1st (29.1%) or 2nd line (44.4%). The 24-month PFS rates were 7.9% in the CT group and 71.2% in the IO group. Using the IPTW method, IO treatment was associated with better PFS (HR=0.227; 95% CI 0.147-0.351; p < 0.0001). The overall response rate was 26.3% in the CT group versus 60.7% in the IO group (p < 0.001) with prolonged duration of disease control in the IO group (p < 0.001). In multivariable analysis, predictive factors of PFS for patients treated with IO were good performance status, absence of liver metastasis and prior primary tumor resection, whereas no association was found for the site of the primary tumor., Conclusions: In the absence of randomized trials, our study highlights the superior efficacy of ICI compared with standard-of-care therapy in patients with unresectable or metastatic dMMR/MSI-H non-colorectal digestive cancer, regardless of tumor type, with acceptable toxicity., Competing Interests: Declaration of Competing Interest AH reports advisory/consultancy fees from Amgen, AstraZeneca, Debiopharm, Eli Lilly and Company, Incyte Corporation, QED Therapeutics. AT has received personal fees from MSD, BMS, Merck Serono, Pierre Fabre, Servier, Incyte Biosciences, Viatris, AstraZeneca. CG has participated in consulting and/or advisory fees for Servier, Sanofi, Merck, has received honoraria as invited speaker from Pierre Fabre, and has received support for travel to meetings from Amgen. DT reports consultancy, advisory fees, honoraria from Servier, Pierre Fabre, Merck Serono, MSD, BMS, AZ, Roche, Sanofi; research funding from Sandoz, AstraZenenca, Servier, MSD; travel grants from Pierre Fabre, MSD, Servier, Roche. EAl reports consultancy and honoraria from Servier, AMGEN, MSD and Merck Serono. EAu reports honoraria from Amgen and Sanofi. FS reports consultancy, advisory fees, honoraria from AMAL Therapeutics, Bayer, BMS, Dragonfly Therapeutics, Merck, Nordic Pharma, Roche, Servier; research funding (institutional) from Amgen, Astra Zeneca, Bayer, BMS, Roche, Sanofi, Travel grants from Amgen, Bayer, Lilly, Servier; and leadership roles at Co-Chair EORTC Task Force Colon, Rectum, Anal Canal. JT has received honoraria as a speaker or in an advisory role from Sanofi, Roche, Merck Serono, Amgen, Servier, Pierre Fabre, Lilly, AstraZeneca and MSD. MBA has received honoraria as a speaker or in an advisory role from BMS, AMGEN, INCYTE, Servier, Pierre Fabre, MercK Serono, Deciphera, Mundipharma, Bayer. RC has received personal fees from Bristol-Myers Squibb, Exeliom Biosciences, Enterome Bioscience, MSD Oncology, Mylan Medical, Pierre Fabre, Servier and non-financial support. RG reports advisory fees from Amgen, AstraZeneca, MSD, Pierre Fabre and travel expenses from BMS. SK reports consultancy, advisory roles, honoraria from Amgen, BeiGene, Boehringer-Ingelheim, Merck, MSD, Pfizer, Servier, and research funding from Behringer-Ingelheim, Bristol-Myers Squibb, Novartis, Roche. SP reports honoraria as a speaker or in an advisory role from Amgen, Pierre Fabre, Servier, BMS, MSD, Merck Serono, Bayer, Sanofi. TAn reports attending advisory board meetings and receiving consulting fees from Astellas, Aptitude Health, Bristol Myers Squibb, Gritstone Oncology, GamaMabs Pharma Sa, Gilead, GlaxoSmithKline, Merck & Co. Inc., Nordic Oncology, Pierre Fabre, Seagen, Servier and Transgène; honoraria from Bristol Myers Squibb, GlaxoSmithKline, Merck & Co. Inc., Merck Serono, Pierre Fabre, Roche Sanofi, Seagen and Servier and support for meetings from Merck & Co. Inc. and Servier. TAp reports honoraria from Amgen, Pierre Fabre, Servier and advisory fees from BMS, MSD, Pierre Fabre. TM reports consultancy, advisory fees, honoraria from Servier, Pierre Fabre, Merck Serono, AAA, Sanofi, and research funding from Amgen; travel grants from Pierre Fabre, Merck Serono, Sanofi. VH reports consultancy, advisory roles, honoraria from AAA, Amgen, Esteve, Ipsen, Deciphera, Servier, Pierre Fabre, Merck Serono. All other authors have declared no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Lynch syndrome (LS) is the most frequent inherited colorectal cancer syndrome., Aim: To assess the burden of adenoma in LS patients under 50 years-old followed in the PRED-IdF network., Methods: From January 2010 to January 2019, all patients under 50 years of age with a confirmed LS germline mutation were included. The main objective was the description of adenomas characteristics according to path&#95;MMR., Results: We analyzed data from 708 patients (mean age 34.8 ± 8.6), of which 41.8 % were male. Among these patients, 37.6% had path&#95;MLH1, 45.4% path&#95;MSH2, 13.9% path&#95;MSH6, 2.9% path&#95;PMS2, and 1.2% path&#95;EpCAM. The analysis included 1721 (70.9%) follow-up colonoscopies. A total of 682 adenomas were detected, including 140 (20.5%) advanced adenomas. The adenoma detection rates during the first and follow-up colonoscopies were 19.2% and 20.5%, respectively. Most adenomas were <10 mm (57.9%), located in the proximal colon (334, 48.9%), and presented as non-polypoid lesions (493, 72.3%). The median growth time for adenomas was 23 months (range 9-114) irrespective of the path&#95;MMR mutation (p = 0.62)., Conclusion: LS patients under 50 years of age have a high burden of adenomas, particularly small non-polypoid adenomas located in the proximal colon. These results highlight the need for intensive screening, with a particular focus on the proximal colon., Competing Interests: Declaration of Competing Interest Authors declare no Conflict of Interests for this article., (Copyright © 2023 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

Systematic screening for pancreatic cancer in high risk individuals is justified by the poor prognosis of the majority of cases diagnosed at a symptomatic stage that are mostly advanced and unresectable Individual risk assessment is based on both genetic data and family history. The screening of a panel of susceptibiility genes should be offered to any affected individual when a genetic predisposition is suspected. An international consortium has proposed a definition of the at risk population, candidate for screening, and there is a consensus on the target lesions of this screening: early adenocarcinoma and benign lesions with a high potential for malignant transformation: Intraductal Papillary Mucinous Neopasm (IPMN) and Pancreatic Intraepithelial Neoplasia (PanIN) with high-grade dysplasia. Its modalities currently consist of an annual pancreatic MRI and/or endoscopic ultrasound (EUS), associated with screening for diabetes mellitus. The main limitation of screening, the effectiveness of which has not yet been demonstrated, is its lack of sensitivity, which results in a non-negligible rate of interval cancers and sometimes advanced diagnoses. Insufficient specificity is also imperfect, in particular with regard to benign lesions with a low potential for degeneration, and can lead to the proposal of unjustified surgeries. This situation makes the future integration of new imaging techniques and promising new biological approaches that are being explored highly desirable., (Copyright © 2023 Société Française du Cancer. Published by Elsevier Masson SAS. All rights reserved.)

Background: The outstanding efficacy of immunotherapy in metastatic dMMR/MSI gastro-intestinal (GI) cancers has led to a rapid increase in the number of patients treated. However, 20-30% of patients experience primary resistance to immune checkpoint inhibitors (ICIPR) and need better characterization., Methods: This AGEO real-world study retrospectively analyzed the efficacy and safety of ICIs and identified clinical variables associated with ICIPR in patients with metastatic dMMR/MSI GI cancers treated with immunotherapy between 2015 and 2022., Results: 399 patients were included, 284 with colorectal cancer (CRC) and 115 with non-CRC, mostly treated by an anti-PD(L)1 (88.0%). PFS at 24 months was 55.8% (95CI [50.8-61.2]) and OS at 48 months was 59.1% (95CI [53.0-65.9]). ORR was 51.0%, and 25.1% of patients were ICIPR. There was no statistical difference in ORR, DCR, PFS, or OS between CRC and non-CRC groups. In multivariable analysis, ICIPR was associated with ECOG-PS ≥ 2 (OR = 3.36), liver metastases (OR = 2.19), peritoneal metastases (OR = 2.00), ≥1 previous line of treatment (OR = 1.83), and age≤50 years old (OR = 1.76)., Conclusion: These five clinical factors associated with primary resistance to ICIs should be considered by physicians to guide treatment choice in GI dMMR/MSI metastatic cancer patients., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

The inner ear is the hub where hair cells (HCs) transduce sound, gravity, and head acceleration stimuli to the brain. Hearing and balance rely on mechanosensation, the fastest sensory signals transmitted to the brain. The mechanoelectrical transducer (MET) channel is the entryway for the sound-balance-brain interface, but the channel-complex composition is not entirely known. Here, we report that the mouse utilizes Piezo1 (Pz1) and Piezo2 (Pz2) isoforms as MET-complex components. The Pz channels, expressed in HC stereocilia, and cell lines are co-localized and co-assembled with MET complex partners. Mice expressing non-functional Pz1 and Pz2 at the ROSA26 locus have impaired auditory and vestibular traits that can only be explained if the Pzs are integral to the MET complex. We suggest that Pz subunits constitute part of the MET complex and that interactions with other MET complex components yield functional MET units to generate HC MET currents., (© 2024. The Author(s).)

Cristae are high-curvature structures in the inner mitochondrial membrane (IMM) that are crucial for ATP production. While cristae-shaping proteins have been defined, analogous lipid-based mechanisms have yet to be elucidated. Here, we combine experimental lipidome dissection with multi-scale modeling to investigate how lipid interactions dictate IMM morphology and ATP generation. When modulating phospholipid (PL) saturation in engineered yeast strains, we observed a surprisingly abrupt breakpoint in IMM topology driven by a continuous loss of ATP synthase organization at cristae ridges. We found that cardiolipin (CL) specifically buffers the inner mitochondrial membrane against curvature loss, an effect that is independent of ATP synthase dimerization. To explain this interaction, we developed a continuum model for cristae tubule formation that integrates both lipid and protein-mediated curvatures. This model highlighted a snapthrough instability, which drives IMM collapse upon small changes in membrane properties. We also showed that cardiolipin is essential in low-oxygen conditions that promote PL saturation. These results demonstrate that the mechanical function of cardiolipin is dependent on the surrounding lipid and protein components of the IMM., (© 2023 The Authors. Published under the terms of the CC BY NC ND 4.0 license.)

Introduction: Almost all patients receiving mechanical ventilation (MV) in intensive care units (ICUs) require analgesia and sedation. The most widely used sedative drug is propofol, but there is uncertainty whether alpha2-agonists are superior. The alpha 2 agonists for sedation to produce better outcomes from critical illness (A2B) trial aims to determine whether clonidine or dexmedetomidine (or both) are clinically and cost-effective in MV ICU patients compared with usual care., Methods and Analysis: Adult ICU patients within 48 hours of starting MV, expected to require at least 24 hours further MV, are randomised in an open-label three arm trial to receive propofol (usual care) or clonidine or dexmedetomidine as primary sedative, plus analgesia according to local practice. Exclusions include patients with primary brain injury; postcardiac arrest; other neurological conditions; or bradycardia. Unless clinically contraindicated, sedation is titrated using weight-based dosing guidance to achieve a Richmond-Agitation-Sedation score of -2 or greater as early as considered safe by clinicians. The primary outcome is time to successful extubation. Secondary ICU outcomes include delirium and coma incidence/duration, sedation quality, predefined adverse events, mortality and ICU length of stay. Post-ICU outcomes include mortality, anxiety and depression, post-traumatic stress, cognitive function and health-related quality of life at 6-month follow-up. A process evaluation and health economic evaluation are embedded in the trial.The analytic framework uses a hierarchical approach to maximise efficiency and control type I error. Stage 1 tests whether each alpha2-agonist is superior to propofol. If either/both interventions are superior, stages 2 and 3 testing explores which alpha2-agonist is more effective. To detect a mean difference of 2 days in MV duration, we aim to recruit 1437 patients (479 per group) in 40-50 UK ICUs., Ethics and Dissemination: The Scotland A REC approved the trial (18/SS/0085). We use a surrogate decision-maker or deferred consent model consistent with UK law. Dissemination will be via publications, presentations and updated guidelines., Trial Registration Number: ClinicalTrials.gov NCT03653832., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY. Published by BMJ.)

Caloric restriction (CR) extends organismal lifespan and health span by improving glucose homeostasis mechanisms. How CR affects organellar structure and function of pancreatic beta cells over the lifetime of the animal remains unknown. Here, we used single nucleus transcriptomics to show that CR increases the expression of genes for beta cell identity, protein processing, and organelle homeostasis. Gene regulatory network analysis link this transcriptional phenotype to transcription factors involved in beta cell identity (Mafa) and homeostasis (Atf6). Imaging metabolomics further demonstrates that CR beta cells are more energetically competent. In fact, high-resolution light and electron microscopy indicates that CR reduces beta cell mitophagy and increases mitochondria mass, increasing mitochondrial ATP generation. Finally, we show that long-term CR delays the onset of beta cell aging and senescence to promote longevity by reducing beta cell turnover. Therefore, CR could be a feasible approach to preserve compromised beta cells during aging and diabetes., Competing Interests: Declaration of Interest. The authors declare no competing interests.

Background: Despite improvements in characterization of CRC heterogeneity, appropriate risk stratification tools are still lacking in clinical practice. This study aimed to elucidate the primary tumor transcriptomic signatures associated with distinct metastatic routes., Methods: Primary tumor specimens obtained from CRC patients with either isolated LM (CRC-Liver) or PM (CRC-Peritoneum) were analyzed by transcriptomic mRNA sequencing, gene set enrichment analyses (GSEA) and immunohistochemistry. We further assessed the clinico-pathological associations and prognostic value of our signature in the COAD-TCGA independent cohort., Results: We identified a significantly different distribution of Consensus Molecular Subtypes between CRC-Liver and CRC-peritoneum groups. A transcriptomic signature based on 61 genes discriminated between liver and peritoneal metastatic routes. GSEA showed a higher expression of immune response and epithelial invasion pathways in CRC-Peritoneum samples and activation of proliferation and metabolic pathways in CRC-Liver samples. The biological relevance of RNA-Seq results was validated by the immunohistochemical expression of three significantly differentially expressed genes ( ACE2, CLDN18 and DUSP4 ) in our signature. In silico analysis of the COAD-TCGA showed that the CRC-Peritoneum signature was associated with negative prognostic factors and poor overall and disease-free survivals., Conclusions: CRC primary tumors spreading to the liver and peritoneum display significantly different transcriptomic profiles. The implementation of this signature in clinical practice could contribute to identify new therapeutic targets for stage IV CRC and to define individualized follow-up programs in stage II-III CRC.

Cristae are high curvature structures in the inner mitochondrial membrane (IMM) that are crucial for ATP production. While cristae-shaping proteins have been defined, analogous mechanisms for lipids have yet to be elucidated. Here we combine experimental lipidome dissection with multi-scale modeling to investigate how lipid interactions dictate IMM morphology and ATP generation. When modulating phospholipid (PL) saturation in engineered yeast strains, we observed a surprisingly abrupt breakpoint in IMM topology driven by a continuous loss of ATP synthase organization at cristae ridges. We found that cardiolipin (CL) specifically buffers the IMM against curvature loss, an effect that is independent of ATP synthase dimerization. To explain this interaction, we developed a continuum model for cristae tubule formation that integrates both lipid and protein-mediated curvatures. The model highlighted a snapthrough instability, which drives IMM collapse upon small changes in membrane properties. We also showed that CL is essential in low oxygen conditions that promote PL saturation. These results demonstrate that the mechanical function of CL is dependent on the surrounding lipid and protein components of the IMM., Competing Interests: Disclosure and Competing Interests Statement The authors declare that they have no conflict of interest.

Some patients with Lynch syndrome (LS) have extreme phenotypes, i.e. cancer before the recommended screening age, or cancer for which there are no screening guidelines. We made the hypothesis that additional germline variants in cancer susceptibility genes (CSG) could explain some of these phenotypes. We compared the prevalence of additional CSG variants in LS patients with a cancer diagnosis before age 30 (early-onset, EO group) and after 40 (usual-onset, UO group). While there was no overall difference, we did find an excess of pathogenic variants and variants of unknown significance in EO cases when only gastrointestinal CSG were considered (OR 2.25; 95% CI: 1.01-5.06, p value = 0.04). Four EO cases stood out: two with POLE/POLD1 variants in the key exonuclease domain, one with a BMPR1A duplication and one with an EPCAM deletion. Additional germline variants should be considered in future screening recommendations, as they might influence cancer risk., (© 2023. The Author(s), under exclusive licence to European Society of Human Genetics.)

The rod photoreceptor synapse is the first synapse of dim-light vision and one of the most complex in the mammalian CNS. The components of its unique structure, a presynaptic ribbon and a single synaptic invagination enclosing several postsynaptic processes, have been identified, but disagreements about their organization remain. Here, we have used EM tomography to generate high-resolution images of 3-D volumes of the rod synapse from the female domestic cat. We have resolved the synaptic ribbon as a single structure, with a single arciform density, indicating the presence of one long site of transmitter release. The organization of the postsynaptic processes, which has been difficult to resolve with past methods, appears as a tetrad arrangement of two horizontal cell and two rod bipolar cell processes. Retinal detachment severely disrupts this organization. After 7 d, EM tomography reveals withdrawal of rod bipolar dendrites from most spherules; fragmentation of synaptic ribbons, which lose their tight association with the presynaptic membrane; and loss of the highly branched telodendria of the horizontal cell axon terminals. After detachment, the hilus, the opening through which postsynaptic processes enter the invagination, enlarges, exposing the normally sequestered environment within the invagination to the extracellular space of the outer plexiform layer. Our use of EM tomography provides the most accurate description to date of the complex rod synapse and details changes it undergoes during outer segment degeneration. These changes would be expected to disrupt the flow of information in the rod pathway. SIGNIFICANCE STATEMENT Ribbon-type synapses transmit the first electrical signals of vision and hearing. Despite their crucial role in sensory physiology, the three-dimensional ultrastructure of these synapses, especially the complex organization of the rod photoreceptor synapse, is not well understood. We used EM tomography to obtain 3-D imaging at nanoscale resolution to help resolve the organization of rod synapses in normal and detached retinas. This approach has enabled us to show that in the normal retina a single ribbon and arciform density oppose a tetrad of postsynaptic processes. In addition, it enabled us to provide a 3-D perspective of the ultrastructural changes that occur in response to retinal detachment., (Copyright © 2023 the authors.)

The inner ear is the hub where hair cells transduce sound, gravity, and head acceleration stimuli carried by neural codes to the brain. Of all the senses, hearing and balance, which rely on mechanosensation, are the fastest sensory signals transmitted to the central nervous system. The mechanoelectrical transducer (MET) channel in hair cells is the entryway for the sound-balance-brain interface, but the channel's composition has eluded biologists due to its complexity. Here, we report that the mouse utilizes Piezo1 (Pz1) and Piezo2 (Pz2) isoforms as central components of the MET complex. The Pz channel subunits are expressed in hair-cell stereocilia, are co-localized and co-assembled, and are essential components of the MET complex in vitro and in situ, including integration with the transmembrane channel (Tmc1/2) protein. Mice expressing non-functional Pz1 and Pz2, but not functional Pz1 at the ROSA26 locus under the control of hair-cell promoters, have impaired auditory and vestibular traits that can only be explained if Pz channel multimers are integral to the MET complex. We affirm that Pz protein subunits constitute MET channels and that functional interactions with components of the MET complex yield current properties resembling hair-cell MET currents. Our results demonstrate Pz is a MET channel component central to interacting with MET complex proteins. Results account for the MET channel pore and complex., Competing Interests: Additional Declarations: There is NO Competing Interest.

Background: Immunotherapy demonstrated remarkable efficacy in metastatic colorectal cancers (mCRCs) with mismatch repair deficiency (MMRd)/microsatellite instability (MSI). However, data regarding efficacy and safety of immunotherapy in the routine clinical practice are scarce., Patients and Methods: This is a retrospective, multicenter study aiming to evaluate efficacy and safety of immunotherapy in routine clinical practice and to identify predictive markers for long-term benefit. Long-term benefit was defined as progression-free survival (PFS) exceeding 24 months. All patients who received immunotherapy for an MMRd/MSI mCRC were included. Patients who received immunotherapy in combination with another known effective therapeutic class agent (chemotherapy or tailored therapy) were excluded., Results: Overall, 284 patients across 19 tertiary cancer centers were included. After a median follow-up of 26.8 months, the median overall survival (mOS) was 65.4 months [95% confidence interval (CI) 53.8 months-not reached (NR)] and the median PFS (mPFS) was 37.9 months (95% CI 30.9 months-NR). There was no difference in terms of efficacy or toxicity between patients treated in the real-world or as part of a clinical trial. Overall, 46.6% of patients had long-term benefit. Independent markers associated with long-term benefit were Eastern Cooperative Oncology Group-performance status (ECOG-PS) 0 (P = 0.025) and absence of peritoneal metastases (P = 0.009)., Conclusions: Our study confirms the efficacy and safety of immunotherapy in patients with advanced MMRd/MSI CRC in the routine clinical practice. ECOG-PS score and absence of peritoneal metastases provide simple markers that could help identify patients who benefit the most from this treatment., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: Telemedicine enthusiasm and uptake in radiation oncology rapidly increased during the COVID-19 pandemic, but it is unclear if and how telemedicine should be used after the COVID-19 public health emergency ends is unclear. Herein, we report on our institution's provider experience after the mature adoption of telemedicine., Methods and Materials: We distributed a survey to all radiation oncology attending physicians at our institution in October 2021 to assess satisfaction, facilitators, and barriers to telemedicine implementation. We performed quantitative and qualitative analyses to characterize satisfaction and identify influencing factors whether telemedicine is employed. We calculated the average proportion of visits that providers expected to be appropriately performed with telemedicine for each disease site and visit type., Results: A total of 60 of the 82 eligible radiation oncologists (73%) responded to the survey, of whom 78% were satisfied with telemedicine in the radiation oncology department and 83% wished to continue offering video visits after the COVID-19 public health emergency ends. Common patient factors influencing whether physicians offer telemedicine include the patient's travel burden, patient preferences, and whether a physical examination is required. Approximately 20% of new consultations and 50% of weekly management visits were estimated to be appropriate for telemedicine. The central nervous system/pediatrics and thoracic faculty considered telemedicine appropriate for the greatest proportion of new consultations, and 93% of respondents felt comfortable determining whether telemedicine was appropriate., Conclusions: Surveyed radiation oncologists were satisfied with telemedicine in their practice, and wished to continue offering video visits in the future. Our data suggest that payers should continue to support this patient-centered technology., (© 2022 The Author(s).)

177 Lu-PSMA is an effective treatment in metastatic castration-resistant prostate cancer (mCRPC). Our ability to assess response rates and adjust treatment may be improved using predictive tools. This study aimed to evaluate change in 177 Lu-PSMA SPECT quantitative parameters to monitor treatment response. Methods: One hundred twenty-seven men with progressive mCRPC previously treated with androgen-signaling inhibition (99%) and chemotherapy (71%) received a median of 3 (interquartile range [IQR], 2-5) 8-GBq (IQR, 8-8.5 GBq) doses of 177 Lu-PSMA-I&T. Imaging included 68 Ga-PSMA-11 PET/CT (SUV max > 15 at a single site and > 10 at all sites > 2 cm), diagnostic CT, and 177 Lu SPECT/CT from vertex to mid thigh (24 h after treatment). 177 Lu SPECT/CT quantitative analysis was undertaken at cycles 1 (baseline) and 2 (week 6) of treatment. Clinical and biochemical results were assessed to evaluate prostate-specific antigen (PSA) progression-free survival (PFS) and overall survival (OS). Results: A PSA reduction of more than 50% was seen in 58% (74/127). The median PSA PFS was 6.1 mo (95% CI, 5.5-6.7), and OS was 16.8 mo (95% CI, 13.5-20.1). At the time of analysis, 41% (52/127) were deceased. At baseline and week 6, 76% (96/127) had analyzable serial 177 Lu SPECT/CT imaging. SPECT total tumor volume (TTV) was reduced between baseline and week 6 in 74% (71/96; median, -193; IQR, -486 to -41). Any increase in SPECT TTV between baseline and week 6 was associated with significantly shorter PSA PFS (hazard ratio, 2.5; 95% CI, 1.5-4.2; P = 0.0008) but not OS. Median PSA PFS in those with an increase in SPECT TTV was 3.7 mo (95% CI, 2.8-6.8), compared with 6.7 mo (95% CI, 5.8-10.6) in those with no increase in SPECT TTV. An increase in SPECT TTV greater than 20% was also associated with PSA PFS (hazard ratio, 1.9; 95% CI, 1.2-3.0; P = 0.008) but less significantly than any change in SPECT TTV. There was a significant difference in PSA PFS between patients with both increased PSA and SPECT TTV and patients with reduced SPECT TTV and PSA (median, 2.8 vs. 9.0 mo; P < 0.0001). Conclusion: Increasing PSMA SPECT TTV on quantitative 177 Lu SPECT/CT predicts short progression-free survival and may play a future role as an imaging response biomarker, identifying when to cease or intensify 177 Lu-PSMA therapy., (© 2023 by the Society of Nuclear Medicine and Molecular Imaging.)

Hypoxia poses a significant energetic challenge and most species exhibit metabolic remodelling when exposed to prolonged hypoxia. One component of this remodelling is mitochondrial biogenesis/mitophagy, which alter mitochondrial abundance and helps to adjust metabolic throughput to match changes in energy demands in hypoxia. However, how acute hypoxia impacts mitochondrial abundance in hypoxia-tolerant species is poorly understood. To help address this gap, we exposed hypoxia-tolerant naked mole-rats to 3 h of normoxia or acute hypoxia (5% O 2 ) and measured changes in mitochondrial abundance using two well-established markers: citrate synthase (CS) enzyme activity and mitochondrial DNA (mtDNA) abundance. We found that neither marker changed with hypoxia in brain, liver, or kidney, suggesting that mitochondrial biogenesis is not initiated during acute hypoxia in these tissues. Conversely in skeletal muscle, the ratio of CS activity to total protein decreased 50% with hypoxia. However, this change was likely driven by an increase in soluble protein density in hypoxia because CS activity was unchanged relative to wet tissue weight and the mtDNA copy number was unchanged. To confirm this, we examined skeletal muscle mitochondria using transmission electron microscopy and found no change in mitochondrial volume density. Taken together with previous studies of mitochondrial respiratory function, our present findings suggest that naked mole-rats primarily rely on tissue-specific functional remodelling of metabolic pathways and mitochondrial respiratory throughput, and not physical changes in mitochondrial number or volume, to adjust to short-term hypoxic exposure., Competing Interests: Declaration of Competing Interest We have no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

In this work, we developed a novel approach to purify [ 11 C]Raclopride ([ 11 C]RAC), an important positron emission tomography radiotracer, based on tailored shape-recognition polymers, with the aim to substitute single-pass HPLC purification with an in-flow trap & release process. Molecular imprinting technology (MIT) applied to solid phase extraction (MISPE) was investigated to develop a setting able to selectively extract [ 11 C]RAC in a mixture containing a high amount of its precursor, (S)-O-Des-Methyl-Raclopride (DM-RAC). Two imprinted polymers selective for unlabeled RAC and DM-RAC were synthesized through a radical polymerization at 65 °C using methacrylic acid and trimethylolpropane trimethacrylate in the presence of template molecule (RAC or DM-RAC). The prepared polymer was characterized by scanning electron microscopy and Fourier transform infrared spectroscopy and tested in MISPE experiments. The polymers were used in testing conditions, revealing a high retention capacity of RAC-MISPE to retain RAC either in the presence of similar concentrations of RAC and DM-RAC precursor (96.9%, RSD 6.6%) and in the presence of a large excess of precursor (90%, RSD 4.6%) in the loading solution. Starting from these promising results, preliminary studies for selective purification of [ 11 C]Raclopride using this RAC-MISPE were performed and, while generally confirming the selectivity capacity of the polymer, revealed challenging applicability to the current synthetic process, mainly due to high backpressures and long elution times.

Some Alphaproteobacteria contain intracytoplasmic membranes (ICMs) and proteins homologous to those responsible for the mitochondrial cristae, an observation which has given rise to the hypothesis that the Alphaproteobacteria endosymbiont had already evolved cristae-like structures and functions. However, our knowledge of microbial fine structure is still limited, leaving open the possibility of structurally homologous ICMs outside the Alphaproteobacteria . Here, we report on the detailed characterization of lamellar cristae-like ICMs in environmental sulfate-reducing Desulfobacterota that form syntrophic partnerships with anaerobic methane-oxidizing (ANME) archaea. These structures are junction-bound to the cytoplasmic membrane and resemble the form seen in the lamellar cristae of opisthokont mitochondria. Extending these observations, we also characterized similar structures in Desulfovibrio carbinolicus, a close relative of the magnetotactic D. magneticus , which does not contain magnetosomes. Despite a remarkable structural similarity, the key proteins involved in cristae formation have not yet been identified in Desulfobacterota , suggesting that an analogous, but not a homologous, protein organization system developed during the evolution of some members of Desulfobacterota . IMPORTANCE Working with anaerobic consortia of methane oxidizing ANME archaea and their sulfate-reducing bacterial partners recovered from deep sea sediments and with the related sulfate-reducing bacterial isolate D. carbinolicus , we discovered that their intracytoplasmic membranes (ICMs) appear remarkably similar to lamellar cristae. Three-dimensional electron microscopy allowed for the novel analysis of the nanoscale attachment of ICMs to the cytoplasmic membrane, and these ICMs are structurally nearly identical to the crista junction architecture seen in metazoan mitochondria. However, the core junction-forming proteins must be different. The outer membrane vesicles were observed to bud from syntrophic Desulfobacterota , and darkly stained granules were prominent in both Desulfobacterota and D. carbinolicus . These findings expand the taxonomic breadth of ICM-producing microorganisms and add to our understanding of three-dimensional microbial fine structure in environmental microorganisms.

Introduction: Sepsis is characterised by a dysregulated immune response to infection, with exaggerated pro-inflammatory and anti-inflammatory responses. A predominant immunosuppressive profile affecting both innate and adaptive immune responses is associated with increased hospital-acquired infection and reduced infection-free survival. While hospital-acquired infection leads to additional antibiotic use, the role of the immunosuppressive phenotype in guiding complex decisions, such as those affecting antibiotic stewardship, is uncertain. This study is a mechanistic substudy embedded within a multicentre clinical and cost-effectiveness trial of biomarker-guided antibiotic stewardship. This mechanistic study aims to determine the effect of sepsis-associated immunosuppression on the trial outcome measures., Methods and Analysis: RISC-sepsis is a prospective, multicentre, exploratory, observational study embedded within the ADAPT-sepsis trial. A subgroup of 180 participants with antibiotics commenced for suspected sepsis, enrolled in the ADAPT-sepsis trial, will be recruited. Blood samples will be collected on alternate days until day 7. At each time point, blood will be collected for flow cytometric analysis into cell preservation tubes. Immunophenotyping will be performed at a central testing hub by flow cytometry. The primary outcome measures are monocyte human leucocyte antigen-DR; neutrophil CD88; programmed cell death-1 on monocytes, neutrophils and T lymphocytes and the percentage of regulatory T cells. Secondary outcome measures will link to trial outcomes from the ADAPT-sepsis trial including antibiotic days; occurrence of hospital-acquired infection and length of ICU-stay and hospital-stay., Ethics and Dissemination: Ethical approval has been granted (IRAS 209815) and RISC-sepsis is registered with the ISRCTN (86837685). Study results will be disseminated by peer-reviewed publications, presentations at scientific meetings and via patient and public participation groups and social media., Competing Interests: Competing interests: DFMcA is NIHR/MRC EME programme director and has previously sat on NIHR HTA funding committees., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY. Published by BMJ.)

Background: The development of vaccines against SARS-CoV2 has been a key public health response to the COVID-19 pandemic. However, since their introduction, there have been reports of anaphylactic reactions to vaccines in individuals with history of allergic reactions to other vaccines, excipients or to COVID vaccines., Aim: A dedicated adult COVID vaccine allergy clinic with a standardised allergy testing protocol was set up to investigate safety and suitability of available COVID vaccines in Australia., Methods: Patients referred to a state-wide COVID-19 vaccine allergy clinic between March and August 2021 with a history of allergy underwent skin-prick testing and intradermal testing to both available vaccine formulations (BNT162b2 and ChAdOx1-S), excipients (polyethylene glycol and polysorbate 80), excipient-containing medications and controls. Basophil activation testing was conducted in few subjects with convincing history of immediate type reactions., Results: Fifty-three patients underwent testing for possible excipient allergy (n = 19), previous non-COVID vaccine reaction (n = 13) or previous reaction to dose 1 of COVID-19 vaccine (n = 21). Patients were predominantly female (n = 43, 81%), aged 18-83 (median 54) years. Forty-four patients tested negative and 42 of these received at least their first dose of a COVID-19 vaccine. Nine patients tested positive to excipients or excipient-containing medication only (n = 3), or vaccines (n = 6). Five patients were positive to just BNT162b2, 3/5 have been vaccinated with ChAdOx1-S. One who was skin test positive to both vaccines, but negative BAT to ChAdOx1-S was successfully vaccinated with ChAdOx1-S., Conclusion: Even in a high-risk population, most patients can be vaccinated with available COVID-19 vaccines. This paper reports local experiences using a combined allergy testing protocol with skin testing and BAT during the pandemic., (© 2022 The Authors. Internal Medicine Journal published by John Wiley & Sons Australia, Ltd on behalf of Royal Australasian College of Physicians.)