Journals
2025 EN
Li Tommy · Gibiansky Leonid · Parikh Apurvasena
+7 more
Epcoritamab is a CD3xCD20 bispecific antibody that activates T cells to kill CD20‐expressing B cells. Epcoritamab is approved for the treatment of adults with different types of relapsed or refractory lymphoma in various geographies, including the United States, Europe, and Japan. Epcoritamab demonstrated an overall response rate of 63%, a complete response rate of 39%, and manageable safety with the approved dosing regimen (0.16‐mg and 0.8‐mg step‐up doses and 48‐mg full dose, with dosing every week in cycles 1–3, every 2 weeks in cycles 4–9, and every 4 weeks in cycles ≥ 10) in patients with relapsed or refractory large B‐cell lymphoma from the phase 1/2 EPCORE® NHL‐1 trial expansion through January 31, 2022. Exposure–efficacy analyses including the EPCORE NHL‐1 and EPCORE NHL‐3 trials revealed that higher exposure was associated with a higher overall response rate, complete response rate, progression‐free survival, and overall survival. A potential plateau of efficacy was observed at 48 mg or above. The exposure–safety analyses of these trials did not identify any safety concerns with the approved dosing regimen. No associations were detected between exposure and safety endpoints. The step‐up doses were clinically active and helped mitigate cytokine release syndrome risk at the subsequent full doses. Most initial responses (94%) were observed during the weekly dosing period, and most responders with large B‐cell lymphoma maintained or improved their response during every 2 weeks and every 4 weeks dosing. Overall, these analyses support the approved single‐agent epcoritamab 0.16/0.8/48‐mg dosing regimen in relapsed or refractory large B‐cell lymphoma.
Journals
2025 EN
Mungai Margaret · Crabtree Amber · Le Han
+26 more
Abstract OPA1 is a dynamin‐related GTPase that modulates mitochondrial dynamics and cristae integrity. Humans carry eight different isoforms of OPA1 and mice carry five, all of which are expressed as short‐ or long‐form isoforms. These isoforms contribute to OPA1's ability to control mitochondrial energetics and DNA maintenance. However, western blot isolation of all long and short isoforms of OPA1 can be difficult. To address this issue, we developed an optimized western blot protocol based on improving running time to isolate five different isoforms of OPA1 in mouse cells and tissues. This protocol can be applied to study changes in mitochondrial structure and function. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol : Western Blot Protocol for Isolating OPA1 Isoforms in Mouse Primary Skeletal Muscle Cells
Journals
2025 EN
Crabtree Amber · Le Han · Harris Chanel
+23 more
Abstract Proximity ligation assays (PLAs) use specific antibodies to detect endogenous protein‐protein interactions. PLAs are a highly useful biochemical technique that allow two proteins within proximity to be visualized with fluorescent probes amplified by PCR. While this technique has gained prominence, the use of a PLA in mouse skeletal muscle (SkM) is novel. In this article, we discuss how the PLA method can be used in SkM to study the protein‐protein interactions within mitochondria‐endoplasmic reticulum contact sites (MERCs). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol : Proximity ligation assay for skeletal muscle tissue and myoblast for MERC proteins
Journals
2025 EN
Jensen Isaac J. · Wells Steven B. · Gras Julien
+1 more
Abstract The majority of immune cells in the human body exist within the tissues rather than in the circulation. Nevertheless, most of our knowledge of the human immune system is biased towards the characterization and understanding of circulating immune cell populations because the latter are readily sampled, whereas cells in tissues are difficult to obtain and/or are limited to single sites of disease. Tissue‐resident cells differ from circulating cells due to tissue‐specific niche adaptations that influence their phenotype and function. For instance, T cells in tissues, resident memory (T RM cells), exhibit tissue‐specific properties that allow optimal protection from infection due to an acquired ability to coordinate rapid and efficacious pathogen clearance. Thus, to fully understand T‐cell responses in various pathological conditions one must focus on the properties of T RM cells and how they have been shaped by their environment. Moreover, one must sample and analyze T cells from multiple tissues, optimally from the same individual, to determine how infectious, cancer, or autoimmune challenge is affecting homeostatic function. Our longstanding collaboration with the organ procurement organization, LiveOnNY, provides unique access to multiple lymphoid, mucosal, and peripheral tissues from organ donors where consent for research use has been obtained. These samples have enabled characterization of human tissue resident memory T cells and other immune cell types across a variety of tissues. Concomitant with this endeavor, we developed and refined a series of methodologies critical for extracting immune cells from tissue for the purpose of phenotypic and mechanistic interrogation. Here, we describe our optimized protocols for processing select human tissues and the requisite coordination and considerations for their maximal yield and tissue quality. © 2025 Wiley Periodicals LLC. Basic Protocol 1 : Isolation of immune cells from blood‐rich sites [spleen (SPL), blood (BLD), bone marrow (BOM)] Basic Protocol 2 : Isolation of immune cells from lymph nodes, tonsils, and thymus [iliac lymph nodes (ILN), lung lymph nodes (LLN), mesenteric lymph nodes (MLN), colonic lymph nodes (CLN), hepatic lymph nodes (HLN), tonsils (TON), thymus (THY)] Basic Protocol 3 : Isolation of immune cells from the lungs [lung (LNG), bronchioalveolar lavage (BAL)] Basic Protocol 4 : Isolation of immune cells from the intestines [jejunum epithelial layer (JEL), jejunum lamina propria (JLP), colon epithelial layer (CEL), colon lamina propria (CLP)] Basic Protocol 5 : Isolation of immune cells from the liver (LVR) Basic Protocol 6 : Immune cell staining for flow cytometry
Journals
2025 EN
Thomas Connor · Hibbard Lainey M. · White Kenneth E.
+1 more
Abstract Skeletal muscle fiber type composition affects muscle function, metabolism, and disease vulnerability. In addition, muscle fiber type analysis informs disease diagnosis and underlying pathophysiology. Multiple methodologies can be used to assess muscle fiber type; however, immunofluorescence (IF) for myosin heavy chain (MyHC) isoforms is the most widely used modern approach due to its relative ease, time‐effectiveness, single‐cell resolution, and capacity to preserve spatial positioning within the native tissue architecture. Here, we present a protocol for IF for MyHC labeling on formalin‐fixed paraffin‐embedded (FFPE) mouse and human muscle sections. We then describe a modified procedure for fiber type analysis of the intact mouse lower hindlimb, enabling high‐throughput muscle composition and morphological analysis across distinct muscles on a single tissue section. Traditionally, IF labeling for MyHC isoforms required fresh tissue flash‐frozen in liquid nitrogen–cooled isopentane, which, while effective, presents challenges for sample processing and preservation, long‐term storage, transport, and biosafety. Comparatively, embedding tissue in paraffin after formalin fixation streamlines clinical workflows, preserves morphology, improves long‐term sample stability, and simplifies sample storage and transport. Furthermore, FFPE effectively inactivates most infectious agents, which can be retained in frozen sections. Thus, FFPE samples are typically safe for standard laboratory handling and are not classified as biohazardous. This approach can be adapted for use with a range of downstream applications, including integration of fiber type analysis with emerging next‐generation techniques that favor FFPE samples. In sum, this method offers a robust alternative to traditional fresh‐frozen protocols and allows for simultaneous fiber type analysis across multiple muscle tissues. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol : Multiplex Immunofluorescence for MyHC Labeling in FFPE Skeletal Muscle Tissues Alternate Protocol 1 : Immunofluorescence for MyHC 2x Labeling in FFPE Skeletal Muscle Tissues Alternate Protocol 2 : Multiplex Immunofluorescence for MyHC Labeling in FFPE Whole Hindlimb Sections
Journals
2025 EN
BowerJernigan Liesl · Anderson Neil O. · Suranyi Robert
+2 more
Abstract The University of Minnesota (UMN) Pyrethrum Breeding Program has been developing Chrysanthemum cinerariifolium with increased levels of Pyrethrin I/Pyrethrin II ratio (%Py) for over a decade. Pyrethrum is an economically important “green pesticide,” producing %Py in glandular trichomes, with the highest concentration in the flowers. The compounds are useful as insecticides for mosquito, deer tick, bed bug, and insects affecting commercial plant production. Selection for early seed germination and flowering in first‐year pyrethrum seedlings successfully selected annualized perennials. The objectives of this research were to determine genetic variation within and among populations of C. cinerariifolium and Chrysanthemum coccineum from commercial seed lots from Africa, Australia, China, South America, the Netherlands, and the United States, as well as a wild population collected in Croatia and resultant F 1 hybrids from the UMN Breeding Program. A total of n = 404 genotypes from 58 populations were tested using DArTseqLD (Diversity Arrays Technology) genotype by sequencing to obtain single nucleotide polymorphism (SNP) markers for 10 different groups. SNP analysis of all taxa resulted in 1786 polymorphic DArTseqLD SNP markers with C. coccineum being genetically distinct from all C. cinerariifolium in the principal component analyses. The global germplasm showed low levels of genetic diversity, demonstrating a very narrow germplasm base and confirmed sharing of genetic stock among private and public sector breeding programs and production firms. We found less genetic variation in the UMN germplasm than expected, although levels of heterozygosity increased in more recent pedigrees. However, several divergent outliers were found as potential sources to enhance genetic variation.
Journals
2025 EN
Denolf Ruben · Doolaege Jordy C. · Selmurzaeva Elina
+7 more
Polymer dissolution‐precipitation recycling is a promising pathway to increase plastic recycling rates. One of the first steps in this process is the dissolution of the plastic, where it is important to understand and predict the dissolution kinetics, specifically for plastic waste. Therefore, the dissolution kinetics of a plasticized poly(vinyl chloride) sample ( P ‐PVC), containing bis(7‐methyloctyl) cyclohexane‐1,2‐dicarboxylate (DINCH), are analyzed in N‐methyl‐2‐pyrrolidinone (NMP), cyclohexanone, methyl ethyl ketone (MEK), and 2‐methyltetrahydrofuran (2‐MeTHF). Additionally, the effects of the particle size (1090–2990 μm) and temperature (30–60 °C) are studied. As expected, lowering the particle size and increasing the temperature reduced the overall dissolution time. It is also found that the dissolution of the polymer and plasticizer occurred simultaneously, and that the fastest dissolution occurred in NMP, followed by cyclohexanone, MEK, and 2‐MeTHF. Finally, both a chain disentanglement model and a first‐order model with a diffusion‐based rate coefficient are fitted to the experimental data to describe the dissolution behavior of the selected plastic waste. The first is found to be a slightly better description of the P ‐PVC dissolution profile in NMP, a known good solvent, while the latter is better at describing the dissolution in MEK and 2‐MeTHF, the slowest analyzed solvents.
Journals
2025 EN
Lower Lillian · Rowland Steven M. · Regula Michael
+7 more
Abstract Battery‐grade graphite and aviation fuel are traditionally produced from non‐renewable, fossil carbon feedstocks and result in substantial greenhouse gas emissions. Biomass holds exciting potential as a renewable and sustainable feedstock for the production of graphite and aviation fuel, but challenges exist including the necessity of a catalyst when producing graphite and low selectivity when producing aviation fuel. A process to convert a biomass‐derived feedstock into graphite without the use of a catalyst and fuels with high selectivity towards sustainable aviation fuel (SAF) is innovated. Heavy bio‐oil undergoes a conversion process similar to the commercial production of synthetic graphite including coking at 500 °C, calcination at 1000 °C, and graphitization at 2800 °C. The resulting biographite exhibits excellent performance in lithium‐ion battery configurations with specific capacity of ~330 mAh g −1 and a 96.8 % capacity rebound after high rate cycling. The liquid hydrocarbon co‐product from coking is suitable for hydrotreating into SAF. The aviation fuel fraction (70 wt % of the fuel produced) meets ASTM standards and is composed primarily of cycloalkanes (~80 wt %) which improves energy density compared to paraffins produced by other SAF pathways and may replace aromatics for elastomer swelling in traditional jet fuel with less soot production.
Journals
2025 EN
Zhou Shengfei · GarcíaMancilla Maximiliano · Francis Clar Jordi
+4 more
Spent liquors of biomass pretreatment provide a source for renewable chemical production. These liquors require treatment before being discharged; otherwise, they negatively impact the environment. Herein, spent liquors from aqueous ammonia pretreatment of poplar wood are characterized for phenolic content via liquid chromatography–mass spectrometry and nuclear magnetic resonance spectroscopy. The main phenolics are phenol, p ‐hydroxybenzamide ( p HBAm), and p ‐hydroxybenzoic acid ( p HBA), of which p HBAm and p HBA are produced from the ester‐linked p ‐hydroxybenzoates in poplar wood. Phenol is produced from p HBA via decarboxylation. The potential biotransformation of the extracted phenolics into 2‐pyrone‐4,6‐dicarboxylic acid (PDC) is assessed using an engineered strain of Novosphingobium aromaticivorans DSM12444 (PDC strain). Biotransformation of p HBAm to PDC is shown to be possible in the presence of p HBA, but not when p HBAm is the sole phenolic substrate, this is the first reported observation of N. aromaticivorans producing PDC from an aromatic amide. The phenol present is not transformed to PDC and does not inhibit PDC production. This study demonstrates that the phenolic amide in spent liquor from ammonia pretreatment can be valorized via biotransformation using N. aromaticivorans , which adds to the growing versatility of N. aromaticivorans as a microbial chassis for converting plant‐derived compounds to useful products.
Journals
2025 EN
Zhou Shengfei · GarcíaMancilla Maximiliano · Francis Clar Jordi
+4 more
The Cover Feature shows microbes engineered at UW‐Madison’s Great Lakes Bioenergy Research Center transforming p ‐hydroxybenzamide ( p HBAm) and p ‐hydroxybenzoic acid ( p HBA) into 2‐pryone‐4,6‐dicarboxylic acid (PDC). Ammonia pretreatment of poplar converts p‐hydroxybenzoate esters into p HBAm, p HBA, and phenol, the structures on the leaves. While phenol is produced by decarboxylation of p HBA, it is not transformed by the microbes. More information can be found in the Research Article by S. D. Karlen and co‐workers (DOI: 10.1002/cssc.202500881 ). Cover design by Chelsea Mamott, Shengfei Zhou and Steven D. Karlen.