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To describe the real-world clinical impact of a commercially available plasma cell-free DNA metagenomic next-generation sequencing assay, the Karius test (KT). We retrospectively evaluated the clinical impact of KT by clinical panel adjudication. Descriptive statistics were used to study associations of diagnostic indications, host characteristics, and KT-generated microbiologic patterns with the clinical impact of KT. Multivariable logistic regression modeling was used to further characterize predictors of higher positive clinical impact. We evaluated 1000 unique clinical cases of KT from 941 patients between January 1, 2017-August 31, 2023. The cohort included adult (70%) and pediatric (30%) patients. The overall clinical impact of KT was positive in 16%, negative in 2%, and no clinical impact in 82% of the cases. Among adult patients, multivariable logistic regression modeling showed that culture-negative endocarditis (OR 2.3; 95% CI, 1.11-4.53; P .022) and concern for fastidious/zoonotic/vector-borne pathogens (OR 2.1; 95% CI, 1.11-3.76; P .019) were associated with positive clinical impact of KT. Host immunocompromised status was not reliably associated with a positive clinical impact of KT (OR 1.03; 95% CI, 0.83-1.29; P .7806). No significant predictors of KT clinical impact were found in pediatric patients. Microbiologic result pattern was also a significant predictor of impact. Our study highlights that despite the positive clinical impact of KT in select situations, most testing results had no clinical impact. We also confirm diagnostic indications where KT may have the highest yield, thereby generating tools for diagnostic stewardship.

Disorganized speech is a critical barrier to recovery in schizophrenia, with profound negative impacts on ones ability to engage with the world. Despite the limited efficacy of existing treatments in addressing disorganization, a qualitative analysis of what leads to disorganization in patient narratives has been lacking. This study addresses this gap through inductive thematic analysis of 30 narrative interviews with individuals with schizophrenia, matched based on whether Formal Thought Disorder (FTD) is present. Through this analysis, we identified four core themes (alienation, interpersonal tension, personal benchmarks, and adverse experiences) and eight subthemes. Our findings suggest that disorganization may serve as a protective mechanism against psychological distress and highlight how the severity of FTD influences these themes. Alienation, particularly due to illness-related stigma, emerged more prominently in those with FTD. The themes of personal benchmarks and interpersonal tension pointed towards a heightened sensitivity to social interactions and self-perception among those with schizophrenia. Adverse experiences, encompassing past challenges, suggest a potential link between trauma and symptom exacerbation. Our qualitative analysis of what themes precede disorganized speech has implications for tailoring psychotherapy. By considering an individuals specific triggers and level of disorganization, therapy may be more effectively targeted to improve recovery-based outcomes. By identifying themes within patient narratives, this study advances our understanding of the qualitative aspects preceding disorganized speech in schizophrenia, paving the way for more personalized and effective recovery-focused interventions.

The pharmaceutical and medicine manufacturing industry has become the largest industrial sector for the employment of chemists, indicating a need for experiments with a pharmaceutical sciences context in the undergraduate chemistry curriculum. In the pharmaceutical industry, testing drug dissolution is a key analytical task for solid oral dosage forms that is performed in different phases of drug development to test the release behavior of new formulations, ensure consistency between manufacturing lots, and help predict the "in vivo" absorption of the drug substance after administration. However, there are a limited number of laboratory experiments in dissolution testing developed for the undergraduate chemistry curriculum. To help students obtain hands-on experience in dissolution testing, a protocol has been developed for an undergraduate chemistry laboratory course for students to build a dissolution apparatus, monitor dissolution processes, model the dissolution to extract kinetic parameters, and evaluate the consistency between dissolution curves with FDA regulated methods. Students successfully collected dissolution curves and completed the modeling analysis with nonlinear least-squares fitting. The designed dissolution protocol has been evaluated to have consistency and reproducibility to be implemented in the undergraduate chemistry laboratory curriculum.

The world population is aging. Comprehensive Geriatric assessment (CGA) has been proven to improve the well-being of older adults. However, evidence suggests not all clinicians implement these recommendations in their practice; nor do all patients adhere to them. Currently, there is no up-to-date review of interventions that can improve older adults' adherence to CGA recommendations and health care provider/clinician implementation of those recommendations. The objective of this scoping review protocol is to describe the methodology of the scoping review with the aim to identify interventions that have been tested to improve adherence to CGA recommendations. We will use the Arksey and O'Malley framework and subsequent extension by Levac and colleagues to complete the scoping review. We searched OVID MEDLINE, OVID Embase, EBSCO CINAHL, APA PsychInfo, and Cochrane CENTRAL databases from inception to November 14, 2024, and will include a review of reference lists of included studies. Studies eligible for inclusion are studies of any design that examined one or more interventions aiming to improve clinician implementation of and patient adherence to CGA in any clinical setting. We will use standard methods for study selection, data abstraction, assessment of methodological quality of individual studies, and data synthesis. Results will be analyzed and reported using descriptive numerical summaries and narrative analysis. Findings from the scoping review will be published in a manuscript and presented at scientific conferences.

Deficient (d) DNA mismatch repair (MMR) is a biomarker predictive of better response to PD-1 blockade immunotherapy in solid tumors. dMMR can be caused by mutations in MMR genes or by protein inactivation, which can be detected by sequencing and immunohistochemistry, respectively. To investigate the role of dMMR in diffuse large B-cell lymphoma (DLBCL), MMR gene mutations and expression of MSH6, MSH2, MLH1, and PMS2 proteins were evaluated by targeted next-generation sequencing and immunohistochemistry in a large cohort of DLBCL patients treated with standard chemoimmunotherapy, and correlated with the tumor immune microenvironment characteristics quantified by fluorescent multiplex immunohistochemistry and gene-expression profiling. The results showed that genetic dMMR was infrequent in DLBCL and was significantly associated with increased cancer gene mutations and favorable immune microenvironment, but not prognostic impact. Phenotypic dMMR was also infrequent, and MMR proteins were commonly expressed in DLBCL. However, intratumor heterogeneity existed, and increased DLBCL cells with phenotypic dMMR correlated with significantly increased T cells and PD-1+ T cells, higher average nearest neighbor distance between T cells and PAX5+ cells, upregulated immune gene signatures, LE4 and LE7 ecotypes and their underlying Ecotyper-defined cell states, suggesting the possibility that increased T cells targeted only tumor cell subsets with dMMR. Only in patients with MYC¯ DLBCL, high MSH6/PMS2 expression showed significant adverse prognostic effects. This study shows the immunologic and prognostic effects of genetic/phenotypic dMMR in DLBCL, and raises a question on whether DLBCL-infiltrating PD-1+ T cells target only tumor subclones, relevant for the efficacy of PD-1 blockade immunotherapy in DLBCL.

In Haiti, manual pit emptiers, known as bayakous, face significant health risks. They work by descending naked into latrine pits, exposing themselves to pathogens and contributing to environmental contamination. This study employs the quantitative microbial risk assessment (QMRA) method to evaluate the microbial risks associated with this practice, considering nine prevalent pathogens in Haiti. Three ingestion scenarios were developed: hand-to-mouth contact, ingestion while immersed in excreta, and a combination of both. A sensitivity analysis assessed the impact of input data on study outcomes. The results indicate a high probability of infection and illness during pit emptying operations annually for all scenarios and pathogens. Recommendations include adopting personal protective equipment (PPE) and using a manual Gulper waste pump to eliminate the need to descend directly into the pits, thereby reducing the risk of injury from sharp objects. The study proposes the establishment of intermediate disposal points approximately 5 km from collection sites to deter illegal dumping. National regulations and professionalization of the bayakou profession are suggested, along with awareness campaigns to promote PPE and Gulper pump usage. Addressing these issues is crucial for safeguarding the health of bayakou and public health in Haiti. HIGHLIGHTS This study is the first scientific QMRA study on manual pit emptiers in Haiti.; It considers nine pathogens and three ingestion scenarios, two of which have not been considered in previous studies.; The results indicate that the emptiers are at higher risk of infection and illness.; It recommends the use of PPE to mitigate health risks and the use of a Gulper pump to eliminate the need to descend directly into pit latrines.;

Low-income tropical regions, such as Haiti, grapple with environmental issues stemming from inadequate sanitation infrastructure for fecal sludge management. This study scrutinizes on-site sanitation systems in these regions, evaluating their environmental impacts and pinpointing improvement opportunities. The focus is specifically on systems integrating excreta valorization through composting and/or anaerobic digestion. Each system encompasses toilet access, evacuation, and sludge treatment. A comparative life cycle assessment was undertaken, with the functional unit managing one ton of excreta in Haiti over a year. Six scenarios representing autonomous sanitation systems were devised by combining three toilet types (container-based toilets (CBTs), ventilated improved pit (VIP) latrines, and flush toilets (WC)) with two sludge treatment processes (composting and biomethanization). Biodigester-based systems exhibited 1.05 times higher sanitary impacts and 1.03 times higher ecosystem impacts than those with composters. Among toilet types, CBTs had the lowest impacts, followed by VIP latrines, with WCs having the highest impacts. On average, WC scenarios were 3.85 times more impactful than VIP latrines and 4.04 times more impactful than those with CBTs regarding human health impact. Critical variables identified include the use of toilet paper, wood shavings, greenhouse gas emissions, and construction materials. HIGHLIGHTS This study analyzes the on-site sanitation chain components throughout their life cycle, assessing three types of toilets in Haiti and two sludge valorization scenarios, resulting in six scenarios.; Scenario 6 had the highest sanitary impact, while Scenario 3 had the lowest.; Scenario 4 showed the highest ecosystem impacts, with Scenario 1 displaying the lowest impact.; Key hotspots identified: toilet paper, wood shavings and GHG emissions.;

Introduction Platelets serve dual roles as a critical component of clot formation and as regulators of immune responses (1, 2). Activated platelets promote inflammation, while circulating resting platelets limit inflammation [...], Lung megakaryocytes (Mks) are largely extravascular with an immune phenotype (1). Because bone marrow (BM) Mks are short lived, it has been assumed that extravascular lung Mks are constantly "seeded" from the BM. To investigate lung Mk origins and how origin affects their functions, we developed methods to specifically label lung Mks using CFSE dye and biotin delivered via the oropharyngeal route. Labeled lung Mks were present for up to 4 months, while BM Mks had a lifespan of less than 1 week. In a parabiosis model, lung Mks were partially replaced over 1 month from a circulating source. Unlike tissue-resident macrophages, using [MDS1.sup.-Cre-ERT2] TdTomato mice, we found that lung Mks arose from hematopoietic stem cells. However, studies with FlkSwitch mTmG mice showed that lung Mks were derived from a Flt3-independent lineage that did not go through a multipotent progenitor. CFSE labeling to track lung Mk-derived platelets showed that approximately 10% of circulating platelets were derived from lung-resident Mks at steady state, but in sterile thrombocytopenia this was doubled (~20%). Lung-derived platelets were similarly increased in a malaria infection model (Plasmodium yoelii) typified by thrombocytopenia. These studies indicate that lung Mks arise from a Flt3- BM source, are long-lived, and contribute more platelets during thrombocytopenia.

Despite therapeutic efficacy observed with immune checkpoint blockade in advanced melanoma, many tumors do not respond to treatment, representing a need for new therapies. Here, we have generated chimeric antigen receptor (CAR) T cells targeting TYRP1, a melanoma differentiation antigen expressed on the surface of melanomas, including rare acral and uveal melanomas. TYRP1-targeted CAR T cells demonstrate antigen-specific activation and cytotoxic activity in vitro and in vivo against human melanomas independent of the MHC alleles and expression. In addition, the toxicity to pigmented normal tissues observed with T lymphocytes expressing TYRP1-targeted TCRs was not observed with TYRP1-targeted CAR T cells. Anti-TYRP1 CAR T cells provide a novel means to target advanced melanomas, serving as a platform for the development of similar novel therapeutic agents and as a tool to interrogate the immunobiology of melanomas.

Recently, screens for mediators of resistance to FLT3 and ABL kinase inhibitors in leukemia resulted in the discovery of LZTR1 as an adapter of a Cullin-3 RING E3 ubiquitin ligase complex responsible for the degradation of RAS GTPases. In parallel, dysregulated LZTR1 expression via aberrant splicing and mutations was identified in clonal hematopoietic conditions. Here we identify that loss of LZTR1, or leukemia-associated mutants in the LZTR1 substrate and RAS GTPase RIT1 that escape degradation, drives hematopoietic stem cell (HSC) expansion and leukemia in vivo. Although RIT1 stabilization was sufficient to drive hematopoietic transformation, transformation mediated by LZTR1 loss required MRAS. Proteolysis targeting chimeras (PROTAC) against RAS or reduction of GTP-loaded RAS overcomes LZTR1 loss-mediated resistance to FLT3 inhibitors. These data reveal proteolysis of noncanonical RAS proteins as novel regulators of HSC self-renewal, define the function of RIT1 and LZTR1 mutations in leukemia, and identify means to overcome drug resistance due to LZTR1 downregulation.SignificanceHere we identify that impairing proteolysis of the noncanonical RAS GTPases RIT1 and MRAS via LZTR1 downregulation or leukemia-associated mutations stabilizing RIT1 enhances MAP kinase activation and drives leukemogenesis. Reducing the abundance of GTP-bound KRAS and NRAS overcomes the resistance to FLT3 kinase inhibitors associated with LZTR1 downregulation in leukemia. This article is highlighted in the In This Issue feature, p. 2221.

Therapies targeting the programmed cell death protein-1/programmed death-ligand 1 (PD-L1) (abbreviated as PD-(L)1) axis are a significant advancement in the treatment of many tumor types. However, many patients receiving these agents fail to respond or have an initial response followed by cancer progression. For these patients, while subsequent immunotherapies that either target a different axis of immune biology or non-immune combination therapies are reasonable treatment options, the lack of predictive biomarkers to follow-on agents is impeding progress in the field. This review summarizes the current knowledge of mechanisms driving resistance to PD-(L)1 therapies, the state of biomarker development along this axis, and inherent challenges in future biomarker development for these immunotherapies. Innovation in the development and application of novel biomarkers and patient selection strategies for PD-(L)1 agents is required to accelerate the delivery of effective treatments to the patients most likely to respond.

Summary: HIV-1 hijacks host proteins involved in membrane trafficking, endocytosis, and autophagy that are critical for virus replication. Molecular details are lacking but are essential to inform on the development of alternative antiviral strategies. Despite their potential as clinical targets, only a few membrane trafficking proteins have been functionally characterized in HIV-1 replication. To further elucidate roles in HIV-1 replication, we performed a CRISPR-Cas9 screen on 140 membrane trafficking proteins. We identified phosphatidylinositol-binding clathrin assembly protein (PICALM) that influences not only infection dynamics but also CD4+ SupT1 biology. The knockout (KO) of PICALM inhibited viral entry. In CD4+ SupT1 T cells, KO cells exhibited defects in intracellular trafficking and increased abundance of intracellular Gag and significant alterations in autophagy, immune checkpoint PD-1 levels, and differentiation markers. Thus, PICALM modulates a variety of pathways that ultimately affect HIV-1 replication, underscoring the potential of PICALM as a future target to control HIV-1.

Immune-checkpoint inhibitors (ICI), although revolutionary in improving long-term survival outcomes, are mostly effective in patients with immune-responsive tumors. Most patients with cancer either do not respond to ICIs at all or experience disease progression after an initial period of response. Treatment resistance to ICIs remains a major challenge and defines the biggest unmet medical need in oncology worldwide. In a collaborative workshop, thought leaders from academic, biopharma, and nonprofit sectors convened to outline a resistance framework to support and guide future immune-resistance research. Here, we explore the initial part of our effort by collating seminal discoveries through the lens of known biological processes. We highlight eight biological processes and refer to them as immune resistance nodes. We examine the seminal discoveries that define each immune resistance node and pose critical questions, which, if answered, would greatly expand our notion of immune resistance. Ultimately, the expansion and application of this work calls for the integration of multiomic high-dimensional analyses from patient-level data to produce a map of resistance phenotypes that can be utilized to guide effective drug development and improved patient outcomes.

I wish to thank you, profoundly, for featuring Palestinian poet Mosab Abu Toha in your November/December 2024 issue. As an activist for Palestine since 2003, and a loyal fan of [...]

Abstract Lymphoma is the sixth most common type of cancer worldwide. Under the current treatment standards, patients with lymphoma often fail to respond to treatment or relapse early and require further therapy. Hence, novel therapeutic strategies need to be explored and our understanding of the molecular underpinnings of lymphomas should be expanded. Ferroptosis, a non-apoptotic regulated cell death, is characterized by increased reactive oxygen species and lipid peroxidation due to metabolic dysfunction. Excessive or lack of ferroptosis has been implicated in tumor development. Current preclinical evidences suggest that ferroptosis participates in tumorigenesis, progression, and drug resistance of lymphoma, identifying a potential biomarker and an attractive molecular target. Our review summarizes the core mechanisms and regulatory networks of ferroptosis and discusses existing evidences of ferroptosis induction for the treatment of lymphoma, with intent to provide a framework for understanding the role of ferroptosis in lymphomagenesis and a new perspective of lymphoma treatment.

A container-based toilet (CBT) is a type of ecological toilet that allows users to compost their feces. During emptying, bucket washing, and composting operations, operators are exposed to microbial risks. This paper aims to evaluate these risks using the Quantitative Microbial Risk Assessment (QMRA) method. Nine pathogens prevalent in Haiti were targeted: Ascaris lumbricoides, Campylobacter spp., Cryptosporidium parvum, Escherichia coli O157:H7, Giardia intestinalis, poliovirus, Salmonella spp., Shigella spp., and Vibrio cholerae. Information regarding pathogens' concentration in feces came from scientific literature data. The exposure scenarios considered were those in which operators accidentally ingested a low dose of feces during the aforementioned operations. A Monte Carlo simulation was conducted to address uncertainties. The results showed that the probability of infection is highly elevated, while the probability of illness is generally moderate or minor, except for poliovirus and Ascaris. Preventive measures can be implemented to reduce these risks during various operations, such as wearing gloves, disposable protective masks, and appropriate clothing. It is up to the political authorities to develop guidelines in this regard and to organize awareness-raising activities with the help of local organizations mandated by the relevant authorities to ensure the safer use of technology by households. HIGHLIGHT Quantitative Microbial Risk Assessment related to container-based toilets (CBTs) in Haïti represents the first scientific QMRA study on composting toilets in the country. It considers nine pathogens, including some that have not been considered in previous studies. Results show the importance of precautions during emptying and composting operations.;

Cette note de lecture résume les principales conclusions d’un mémoire de maîtrise portant sur la relation pédagogique en formation à distance (FAD). Les résultats de cette recherche démontrent qu’en contexte de FAD, bien que plus de la moitié du corps enseignant (n = 73) considère la relation comme satisfaisante, la majorité estime que la distance impacte négativement cette dynamique. Confrontées aux défis liés à la distance physique ou aux limitations techniques, les personnes enseignantes déploient diverses stratégies innovantes pour favoriser un climat de classe positif, une présence accrue et une qualité relationnelle optimale. Les résultats de cette recherche soulèvent des questions sur la FAD, soulignant l’importance d’ajuster les pratiques éducatives tout en préservant la qualité de la relation pédagogique.