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Abstract This study focuses on refining growth-rate-based drug response metrics for patient-derived tumor organoid screening using brightfield live-cell imaging. Traditional metrics like Normalized Growth Rate Inhibition (GR) and Normalized Drug Response (NDR) have been used to assess organoid responses to anticancer treatments but face limitations in accurately quantifying cytostatic and cytotoxic effects across varying growth rates. Here, we introduce the Normalized Organoid Growth Rate (NOGR) metric, specifically developed for brightfield imaging-based assays. A label-free image analysis model was applied to segment organoids precisely, track their growth rates over time, and classify viable and dead organoids. Testing eleven phenotypically distinct pancreatic cancer organoid models with five chemotherapeutics demonstrates that the NOGR metric more effectively captures cytostatic and cytotoxic drug effects compared to existing methods. This approach enhances the biological relevance of drug sensitivity assessments on organoids and offers a valuable tool for advancing personalized cancer treatment strategies.

Abstract Realizing genetic circuits on single DNA molecules as self-encoded dissipative nanodevices is a major step toward miniaturization of autonomous biological systems. A circuit operating on a single DNA implies that genetically encoded proteins localize during coupled transcription-translation to DNA, but a single-molecule measurement demonstrating this has remained a challenge. Here, we use a genetically encoded fluorescent reporter system with improved temporal resolution and observe the synthesis of individual proteins tethered to a DNA molecule by transient complexes of RNA polymerase, messenger RNA, and ribosome. Against expectations in dilute cell-free conditions where equilibrium considerations favor dispersion, these nascent proteins linger long enough to regulate cascaded reactions on the same DNA. We rationally design a pulsatile genetic circuit by encoding an activator and repressor in feedback on the same DNA molecule. Driven by the local synthesis of only several proteins per hour and gene, the circuit dynamics exhibit enhanced variability between individual DNA molecules, and fluctuations with a broad power spectrum. Our results demonstrate that co-expressional localization, as a nonequilibrium process, facilitates single-DNA genetic circuits as dissipative nanodevices, with implications for nanobiotechnology applications and artificial cell design.

Abstract Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal diseases, characterized by a treatment-resistant and invasive nature. In line with these inherent aggressive characteristics, only a subset of patients shows a clinical response to the standard of care therapies, thereby highlighting the need for a more personalized treatment approach. In this study, we comprehensively unraveled the intra-patient response heterogeneity and intrinsic aggressive nature of PDAC on bulk and single-organoid resolution. We leveraged a fully characterized PDAC organoid panel (N = 8) and matched our artificial intelligence-driven, live-cell organoid image analysis with retrospective clinical patient response. In line with the clinical outcomes, we identified patient-specific sensitivities to the standard of care therapies (gemcitabine-paclitaxel and FOLFIRINOX) using a growth rate-based and normalized drug response metric. Moreover, the single-organoid analysis was able to detect resistant as well as invasive PDAC organoid clones, which was orchestrates on a patient, therapy, drug, concentration and time-specific level. Furthermore, our in vitro organoid analysis indicated a correlation with the matched patient progression-free survival (PFS) compared to the current, conventional drug response readouts. This work not only provides valuable insights on the response complexity in PDAC, but it also highlights the potential applications (extendable to other tumor types) and clinical translatability of our approach in drug discovery and the emerging era of personalized medicine.

BackgroundDiabetes affects millions of people worldwide, making them more vulnerable to infections, including seasonal influenza. It is therefore particularly important for those suffering from diabetes to be vaccinated against influenza each year. However, influenza vaccination coverage remains low in this population. This review primarily aims to identify the determinants of influenza vaccination in people with diabetes (T1D or T2D). Secondly, it aims to assess main recommendations for influenza vaccination, vaccine effectiveness, vaccination coverage, and how education and pharmacists can encourage uptake of the vaccine in the diabetic population.MethodsA scoping review was conducted in January 2022 to systematically review evidence on influenza vaccination in people with diabetes using data from PubMed, Science Direct, and EM Premium with terms such as “Diabetes mellitus,” “Immunization Programs,” “Vaccination,” and “Influenza Vaccines.” Quality assessment and data extraction were independently conducted by two authors. Disagreements between the authors were resolved through discussion and consensus, and if necessary, by consulting a third author.ResultsOf the 333 records identified, 55 studies met the eligibility criteria for inclusion in this review. Influenza vaccination was recommended for people ≥6 months. Despite effectiveness evidence showing a reduction in mortality and hospitalizations in people with diabetes vaccinated vs. non-vaccinated ones, very few studies reported a coverage rate ≥ 75%, which is WHO’s target objective. Determinants such as advanced age, presence of comorbidities and healthcare givers’ advice were associated with increased vaccination uptake. On the contrary, fear of adverse reactions and concerns about vaccine effectiveness were significant barriers. Finally, education and pharmacists’ intervention played a key role in promoting vaccination and increasing vaccination uptake.ConclusionInfluenza vaccination coverage in people with diabetes remains low despite recommendations and evidence on vaccine effectiveness. Motivators and barriers as well as several socio-demographic and clinical factors have been identified to explain this trend. Efforts are now needed to increase the number of diabetics vaccinated against influenza, mainly through education and the involvement of healthcare givers.

Abstract Background The purpose of this study was to investigate how feedback and self-assessment strategies affect performance and retention of manual skills in a group of chiropractic students. Methods Seventy-five students participated in two spinal manipulation (SM) learning sessions using a force-sensing table. They were recruited between May and November 2022 during HVLA technical courses. Students were randomly assigned into three different groups: participants in group 1 received visual feedback, those in group 2 received visual feedback after self-assessment, and participants in group 3 (C) received no feedback. During the first session, participants started with one block of 3 familiarization trials, followed by two blocks of 6 SM HVLA (high velocity low amplitude) posterior-to-anterior thoracic SM trials, with 3 trials performed with a target force of 450 N and 3 others at 800 N. They received feedback according to their group during the first block, but no feedback was provided during the second block. All participants were invited to participate in a second session for the retention test and to perform a new set SM without any form of feedback. Results Results showed that visual feedback and visual feedback in addition to self-assessment did not improve short-term SM performance, nor did it improve performance at the one-week retention test. The group that received visual feedback and submitted to self-assessment increased the difference between the target force and the peak force applied, which can be considered a decrease in performance. Conclusion No learning effects between the three groups of students exposed to different feedback and self-assessment learning strategies were highlighted in the present study. However, future research on innovative motor learning strategies could explore the role of external focus of attention, self-motivation and autonomy in SM performance training.

ObjectivesThe objectives were (1) to describe and compare headache-related clinical features between teleworkers with migraine and those with tension-type headache (TTH) and (2) to determine the association between coping strategies and headache frequency, and intensity in the context of the COVID-19 pandemic.MethodsThis cross-sectional online survey was conducted with 284 teleworkers (127 with migraine and 157 with TTH). Sociodemographic data, information related to work factors, headache clinical features, coping strategies used during the COVID-19 pandemic, and headache-related clinical features were compared between headache profiles. Bivariate logistic regression analyses were used to determine the association between coping strategies and headache frequency, and intensity.ResultsResults showed that teleworkers with migraine had longer and more painful headache episodes than teleworkers with TTH (ps

Despite the recent emergence of immune checkpoint inhibitors, clinical outcomes of metastatic NSCLC patients remain poor, pointing out the unmet need to develop novel therapies to enhance the anti-tumor immune response in NSCLC. In this regard, aberrant expression of the immune checkpoint molecule CD70 has been reported on many cancer types, including NSCLC. In this study, the cytotoxic and immune stimulatory potential of an antibody-based anti-CD70 (aCD70) therapy was explored as single agent and in combination with docetaxel and cisplatin in NSCLC in vitro and in vivo. Anti-CD70 therapy resulted in NK-mediated killing of NSCLC cells and increased production of pro-inflammatory cytokines by NK cells in vitro. The combination of chemotherapy and anti-CD70 therapy further enhanced NSCLC cell killing. Moreover, in vivo findings showed that the sequential treatment of chemo-immunotherapy resulted in a significant improved survival and delayed tumor growth compared to single agents in Lewis Lung carcinoma-bearing mice. The immunogenic potential of the chemotherapeutic regimen was further highlighted by increased numbers of dendritic cells in the tumor-draining lymph nodes in these tumor-bearing mice after treatment. The sequential combination therapy resulted in enhanced intratumoral infiltration of both T and NK cells, as well as an increase in the ratio of CD8+ T cells over Tregs. The superior effect of the sequential combination therapy on survival was further confirmed in a NCI-H1975-bearing humanized IL15-NSG-CD34+ mouse model. These novel preclinical data demonstrate the potential of combining chemotherapy and aCD70 therapy to enhance anti-tumor immune responses in NSCLC patients.

After the establishment of a causal relationship between tobacco use and cancer in the 1950s, cellulose acetate filters were introduced with the claim to reduce the adverse health impact of unfiltered cigarettes. Often perceived to be more pleasant and healthy, filters encouraged smoking. However, filtered cigarettes are more deeply inhaled to obtain the same nicotine demand while altered combustion releases more tobacco-specific nitrosamines. The increasing use of cigarette filter ventilation is associated with a sharp rise in lung adenocarcinomas in recent decades. While not preventing adverse health effects, a global environmental problem has been created due to the non-biodegradable filter litter, causing ecotoxicological effects and the spread of microplastics. Recently, the Belgian Superior Health Council advised policymakers to ban cigarette filters as single-use plastics at both national and European levels. This article outlines the arguments used to justify this plea (human health and environment), the expected effects of a filter ban, as well as the public reception and reactions of the tobacco industry. The specific context of the European Union is discussed including the revision of the Single-Use Plastics Directive, affording a new opportunity to ban plastic filters. This perspective article aims to fuel the momentum and cooperation among member states for this purpose.

Abstract Cellular senescence is a state of stable cell-cycle arrest with secretory features in response to cellular stress. Historically, it has been considered as an endogenous evolutionary homeostatic mechanism to eliminate damaged cells, including damaged cells which are at risk of malignant transformation, thereby protecting against cancer. However, accumulation of senescent cells can cause long-term detrimental effects, mainly through the senescence-associated secretory phenotype, and paradoxically contribute to age-related diseases including cancer. Besides its role as tumor suppressor, cellular senescence is increasingly being recognized as an in vivo response in cancer patients to various anticancer therapies. Its role in cancer is ambiguous and even controversial, and senescence has recently been promoted as an emerging hallmark of cancer because of its hallmark-promoting capabilities. In addition, the prognostic implications of cellular senescence have been underappreciated due to the challenging detection and sparse in and ex vivo evidence of cellular senescence in cancer patients, which is only now catching up. In this review, we highlight the approaches and current challenges of in and ex vivo detection of cellular senescence in cancer patients, and we discuss the prognostic implications of cellular senescence based on in and ex vivo evidence in cancer patients.

The article focuses on the articulation of resources at different scales in the processes of reterritorialization of food. Eleven contrasting cases of collective actions are analyzed and compared. Two points are highlighted : reterritorialization relies on the mobilization and creation of territorial resources, which can be of different kinds ; it relies on territorial and extra-territorial resources, the latter being mobilized to initiate and sustain the action, to reinforce it, or to allow it to increase its scope. In particular, these results highlight the role of a multiplicity of actors in the transitions towards a more territorialized food system.

Abstract The immune checkpoint molecule CD70 and its receptor CD27 are aberrantly expressed in many hematological and solid malignancies. Dysregulation of the CD70-CD27 axis within the tumor and its microenvironment is associated with tumor progression and immunosuppression. This is in contrast to physiological conditions, where tightly controlled expression of CD70 and CD27 plays a role in co-stimulation in immune responses. In hematological malignancies, cancer cells co-express CD70 and CD27 promoting stemness, proliferation and survival of malignancy. In solid tumors, only expression of CD70 is present on the tumor cells which can facilitate immune evasion through CD27 expression in the tumor microenvironment. The discovery of these tumor promoting and immunosuppressive effects of the CD70-CD27 axis has unfolded a novel target in the field of oncology, CD70. In this review, we thoroughly discuss current insights into expression patterns and the role of the CD70-CD27 axis in hematological and solid malignancies, its effect on the tumor microenvironment and (pre)clinical therapeutic strategies.

Abstract Associated factors of back pain (BP) development before puberty and its persistence are poorly documented. We investigated the association and possible temporality between prior BP history (PBPH), muscular endurance (ME), aerobic capacity (AC), sport activity variables (SAV) and BP in children aged 6 to 12. We collected baseline characteristics (demographics, PBPH, ME, AC and SAV) of children from three primary schools in Canada. Parents replied to weekly text messages regarding their children BP status over an 8-month period. Logistic regression models were adjusted for potential confounders. Data from 242 children (46% female; 8.6 ± 1.7 years) were included. Over the 8-month survey BP prevalence was 48.1%, while the cumulative incidence was 31.9%. The occurrence of at least one BP event was associated with PBPH [OR (IC 95%) = 6.33 (2.35–17.04)] and high AC [2.89 (1.21–6.90)]. High AC was also associated with the development of a first BP episode [2.78 (1.09–7.07)], but ME and SAV were not. BP appears to be relatively common before puberty. BP history seems to be strongly associated with BP recurrence in children. Aerobic capacity is associated with first BP episode development.

Une récente expérience a été menée dans le cadre d’un travail de terrain d’une promotion d’étudiants. Le projet alimentaire territorial (PAT) initié par le PNR Queyras s’appuie sur une diversité d’acteurs, têtes de réseaux tant institutionnels qu’associatifs. En complément, les étudiants sont allés à la rencontre des habitants du territoire Guillestrois-Queyras, pour connaitre leurs pratiques d’approvisionnement alimentaire et leurs visions de l’agriculture de proximité (ACTERRA, Bletterie et Lardon, 2019). L’enjeu était double, recueillir les habitudes alimentaires des habitants et tester une méthodologie généralisable ensuite sur l’ensemble du territoire du PAT. Le nouveau regard porté sur ce territoire, qui combine un espace urbain autour de la petite ville de Guillestre et un espace rural de la montagne du Queyras, a mis en évidence la dimension fédératrice de l’alimentation et la façon dont une organisation socio-spatiale peut faire « commun » (Icéri et Lardon, 2019).

Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumors that retain their poor prognosis despite recent advances in their standard of care. As the involvement of the immune system against HNSCC development is well-recognized, characterization of the immune signature and the complex interplay between HNSCC and the immune system could lead to the identification of novel therapeutic targets that are required now more than ever. In this study, we investigated RNA sequencing data of 530 HNSCC patients from The Cancer Genome Atlas (TCGA) for which the immune composition (CIBERSORT) was defined by the relative fractions of 10 immune-cell types and expression data of 45 immune checkpoint ligands were quantified. This initial investigation was followed by immunohistochemical (IHC) staining for a curated selection of immune cell types and checkpoint ligands markers in tissue samples of 50 advanced stage HNSCC patients. The outcome of both analyses was correlated with clinicopathological parameters and patient overall survival. Our results indicated that HNSCC tumors are in close contact with both cytotoxic and immunosuppressive immune cells. TCGA data showed prognostic relevance of dendritic cells, M2 macrophages and neutrophils, while IHC analysis associated T cells and natural killer cells with better/worse prognostic outcome. HNSCC tumors in our TCGA cohort showed differential RNA over- and underexpression of 28 immune inhibitory and activating checkpoint ligands compared to healthy tissue. Of these, CD73, CD276 and CD155 gene expression were negative prognostic factors, while CD40L, CEACAM1 and Gal-9 expression were associated with significantly better outcomes. Our IHC analyses confirmed the relevance of CD155 and CD276 protein expression, and in addition PD-L1 expression, as independent negative prognostic factors, while HLA-E overexpression was associated with better outcomes. Lastly, the co-presence of both (i) CD155 positive cells with intratumoral NK cells; and (ii) PD-L1 expression with regulatory T cell infiltration may hold prognostic value for these cohorts. Based on our data, we propose that CD155 and CD276 are promising novel targets for HNSCC, possibly in combination with the current standard of care or novel immunotherapies to come.

This study focuses on refining growth-rate-based drug response metrics for patient-derived tumor organoid screening using brightfield live-cell imaging. Traditional metrics like Normalized Growth Rate Inhibition (GR) and Normalized Drug Response (NDR) have been used to assess organoid responses to anticancer treatments but face limitations in accurately quantifying cytostatic and cytotoxic effects across varying growth rates. Here, we introduce the Normalized Organoid Growth Rate (NOGR) metric, specifically developed for brightfield imaging-based assays. A label-free image analysis model was applied to segment organoids precisely, track their growth rates over time, and classify viable and dead organoids. Testing eleven phenotypically distinct pancreatic cancer organoid models with five chemotherapeutics demonstrates that the NOGR metric more effectively captures cytostatic and cytotoxic drug effects compared to existing methods. This approach enhances the biological relevance of drug sensitivity assessments on organoids and offers a valuable tool for advancing personalized cancer treatment strategies. Introducing the Normalized Organoid Growth Rate (NOGR), a novel drug response metric enhancing the assessment of cytostatic and cytotoxic effects in tumor organoids using brightfield imaging-based assays. [ABSTRACT FROM AUTHOR]

Abstract Background The current sanitary crisis brought on by the COVID-19 recently forced a large proportion of workers to adopt telecommuting with limited time to plan transition. Given that several work-related risk factors are associated with headache and neck pain, it seems important to determine those associated with headache and neck pain in telecommuters. The main objective of this study was to identify which telecommuting and individual associated factors are related with headache and neck pain occurrence in telecommuters over a five days follow-up. The second objective was to evaluate the impact of wearing a headset on headache and neck pain intensity in telecommuters. Methods One hundred and sixty-two participants in telecommuting situation were recruited. Baseline assessment included sociodemographic data, headache and neck pain-related disability (6-item Headache Impact Test (HIT-6) and Neck Bournemouth Questionnaire (NBQ)), headache and neck pain frequency and intensity as well as questions about the wearing of a headset (headset wearing, headset type and headset wearing hours). A prospective data collection of headache, neck pain and headset wearing was conducted using daily e-mail over a 5-day follow-up. A stepwise multivariate regression model was performed to determine associated factors of headache or neck pain occurrence during the follow-up. A t-test was conducted to assess the impact of headset wearing on headache and neck pain intensity during the follow-up. Results Regarding headache, the stepwise multivariate regression model showed that the HIT-6 score was associated with future headache occurrence in telecommuters (OR (95% CI) = 1.094 (1.042–1.148); R 2 = 0.094; p

Abstract Immunotherapy is currently under intensive investigation as a potential breakthrough treatment option for glioblastoma. Given the anatomical and immunological complexities surrounding glioblastoma, lymphocytes that infiltrate the brain to develop durable immunity with memory will be key. Polyinosinic:polycytidylic acid, or poly(I:C), and its derivative poly-ICLC could serve as a priming or boosting therapy to unleash lymphocytes and other factors in the (immuno)therapeutic armory against glioblastoma. Here, we present a systematic review on the effects and efficacy of poly(I:C)/poly-ICLC for glioblastoma treatment, ranging from preclinical work on cellular and murine glioblastoma models to reported and ongoing clinical studies. MEDLINE was searched until 15 May 2021 to identify preclinical (glioblastoma cells, murine models) and clinical studies that investigated poly(I:C) or poly-ICLC in glioblastoma. A systematic review approach was conducted according to PRISMA guidelines. ClinicalTrials.gov was queried for ongoing clinical studies. Direct pro-tumorigenic effects of poly(I:C) on glioblastoma cells have not been described. On the contrary, poly(I:C) changes the immunological profile of glioblastoma cells and can also kill them directly. In murine glioblastoma models, poly(I:C) has shown therapeutic relevance as an adjuvant therapy to several treatment modalities, including vaccination and immune checkpoint blockade. Clinically, mostly as an adjuvant to dendritic cell or peptide vaccines, poly-ICLC has been demonstrated to be safe and capable of eliciting immunological activity to boost therapeutic responses. Poly-ICLC could be a valuable tool to enhance immunotherapeutic approaches for glioblastoma. We conclude by proposing several promising combination strategies that might advance glioblastoma immunotherapy and discuss key pre-clinical aspects to improve clinical translation.

Abstract Melanoma remains a deadly cancer despite significant advances in immune checkpoint blockade and targeted therapies. The incidence of melanoma is also growing worldwide, which highlights the need for novel treatment options and strategic combination of therapies. Here, we investigate non‐thermal plasma (NTP), an ionized gas, as a promising, therapeutic option. In a melanoma mouse model, direct treatment of tumors with NTP results in reduced tumor burden and prolonged survival. Physical characterization of NTP treatment in situ reveals the deposited NTP energy and temperature associated with therapy response, and whole transcriptome analysis of the tumor identified several modulated pathways. NTP treatment also enhances the cancer‐immunity cycle, as immune cells in both the tumor and tumor‐draining lymph nodes appear more stimulated to perform their anti‐cancer functions. Thus, our data suggest that local NTP therapy stimulates systemic, anti‐cancer immunity. We discuss, in detail, how these fundamental insights will help direct the translation of NTP technology into the clinic and inform rational combination strategies to address the challenges in melanoma therapy.

Reversible protein phosphorylation is a post-translational modification involved in virtually all plant processes, as it mediates protein activity and signal transduction. Here, we probe dynamic protein phosphorylation during de novo shoot organogenesis in Arabidopsis thaliana. We find that application of three kinase inhibitors in various time intervals has different effects on root explants. Short exposures to the putative histidine (His) kinase inhibitor TCSA during the initial days on shoot induction medium (SIM) are detrimental for regeneration in seven natural accessions. Investigation of cytokinin signaling mutants, as well as reporter lines for hormone responses and shoot markers, suggests that TCSA impedes cytokinin signal transduction via AHK3, AHK4, AHP3, and AHP5. A mass spectrometry-based phosphoproteome analysis further reveals profound deregulation of Ser/Thr/Tyr phosphoproteins regulating protein modification, transcription, vesicle trafficking, organ morphogenesis, and cation transport. Among TCSA-responsive factors are prior candidates with a role in shoot apical meristem patterning, such as AGO1, BAM1, PLL5, FIP37, TOP1ALPHA, and RBR1, as well as proteins involved in polar auxin transport (e.g., PIN1) and brassinosteroid signaling (e.g., BIN2). Putative novel regeneration determinants regulated by TCSA include RD2, AT1G52780, PVA11, and AVT1C, while NAIP2, OPS, ARR1, QKY, and aquaporins exhibit differential phospholevels on control SIM. LC–MS/MS data are available via ProteomeXchange with identifier PXD030754.

Des regards croisés France-Argentine sont riches d’enseignement pour la formation, mais aussi pour une réflexion méthodologique sur le développement durable des territoires. Ils mettent ici en évidence l’intégration des enjeux et la coexistence de modèles de développement agricoles au niveau local. Dans cas de Chascomús, une ville moyenne située à 150 km au Sud de Buenos Aires, les dynamiques territoriales à l’oeuvre ont été étudiées à travers une recherche-formation-action au cours de laquelle les acteurs locaux se sont projetés à 20 ans, avec des pistes d’actions à cinq ans reliées aux enjeux qui ont été identifiés à l’aide de l’approche spatiale. Nous montrons ici que la démarche participative de Jeu de Territoire, qui a été élaborée en France, peut s’adapter en Argentine et qu’elle contribue efficacement à la formation des étudiants au développement et à l’aménagement des territoires (MS ACTERRA) en France, tout autant qu’à l’apprentissage des collègues chercheurs et formateurs argentins (Réseau AgriteRRIs) et des acteurs locaux (en particulier nos partenaires : l’Agence locale de développement agricole de l’INTA et la Municipalité de Chascomús).