Bruker (United States)

Nuclear magnetic resonance (NMR) spectroscopy is widely adopted for assessing biochemical composition in agriculture. This study evaluated the feasibility of 400 MHz NMR to detect biochemical differences in Hass avocados grown under conventional (N = 101) and regenerative (N = 105) farming practices in Southwestern Australia. Phosphite, associated with Phytophthora root rot management, was a key discriminating feature (area under ROC curve = 0.96), being detected in 90% of conventional avocados (mean: 49 mg/kg) and 6 regenerative samples (mean: 24 mg/kg). To assess translational potential, water extracts of five samples were analyzed using 80 MHz benchtop NMR. Phosphite was detectable below the strictest maximum residue limit (25 mg/kg), demonstrating the potential of NMR as a sustainable and cost-effective solution for monitoring phosphite residues. This proof-of-concept benchtop NMR approach demonstrates analytical feasibility but requires further validation before application in field-based traceability or regulatory contexts, with a potential future relevance to environmental monitoring, sustainable agriculture, and other crop systems.

Using spatially-resolved proteomics via mass spectrometry imaging on fracture callus tissues from young and aged mice, we observed delayed healing in aged animals based on the composition of the extracellular matrices. Higher levels of bone specific collagens were detected in young animals, whereas cartilage specific collagens were detected in aged animals at higher levels. Further, detection of novel, non-canonical callus proteins revealed critical transitional steps that are delayed in aged-callus tissues, and these may also contribute to the delayed healing aged animals.

The research note elaborates on the role of green marketing in creating the hotel brand experience (HBE). It aims to reduce the gap in understanding how hotel brand managers stage green encounters within the brand experience and what they believe drives negative vs. positive guest experiences when exposed to specific green marketing initiatives. The theoretical background is followed by the presentation of results from qualitative research conducted with managers in Croatia's leading four- and five-star hotel chains and standalone hotels. Hotel managers consider green marketing a crucial driver for success and long-term survival. Despite a shared belief that delivering a superior experience is a priority, green initiatives are often chosen primarily on the basis of implementation costs. Encounters in which guests are presented with higher investments by the hotel lead to more positive brand experiences. Although hotel managers believe that affective experiences are the most important, they are particularly interested in evoking cognitive experiences through green marketing messages. Therefore, further research should focus on identifying ways to evoke an affective brand experience.

Spatially mapping the transcriptome and proteome in the same tissue section can profoundly advance our understanding of cellular heterogeneity and function. Here we present Deterministic Barcoding in Tissue sequencing plus (DBiTplus), an integrative multimodal spatial omics approach combining sequencing-based spatial transcriptomics and multiplexed protein imaging on the same section, enabling both single-cell-resolution cell typing and transcriptome-wide interrogation of biological pathways. DBiTplus utilizes spatial barcoding and RNase H-mediated cDNA retrieval, preserving tissue architecture for multiplexed protein imaging. We developed computational pipelines to integrate these modalities, allowing imaging-guided deconvolution to generate single-cell-resolved spatial transcriptome atlases. We demonstrate DBiTplus across diverse samples including frozen mouse embryos, and formalin-fixed paraffin-embedded human lymph nodes and lymphoma tissues, highlighting its compatibility with challenging clinical specimens. DBiTplus uncovered mechanisms of lymphomagenesis, progression and transformation in human lymphomas. Thus, DBiTplus is a unified workflow for spatially resolved single-cell atlasing and unbiased exploration of biological mechanisms in a cell-by-cell manner at transcriptome scale.

ABSTRACT The analytical comparability of biologic products and their biosimilars, including higher-order structure (HOS) assessment, ensures product quality and is required for regulatory approval. In this study, nuclear magnetic resonance (NMR) spectroscopy was used to evaluate the HOS of Humira (adalimumab) and its biosimilars under normal and photo-stressed conditions. Under normal conditions, 1D and 2D NMR spectra showed strong structural similarity among all products. However, photo-stressed samples exhibited distinct structural differences, including increased methionine oxidation, and localized conformational changes, most notably in the reference product. These changes correlated with findings from size-exclusion chromatography, capillary isoelectric focusing, and mass spectrometry (MS), which revealed size and charge heterogeneity, as well as site-specific methionine oxidation in the heavy chains. The differences in photostability were found to be influenced by container closure systems (CCSs) and formulations. In contrast, circular dichroism spectral analysis showed minimal variation in secondary structures among stressed and unstressed samples. These results underscore the utility of NMR as a sensitive tool for comparative structural analysis of monoclonal antibodies and their biosimilars, particularly under stress conditions, and highlight the impact of formulation and CCS on product stability.

Complex NMR process monitoring often requires chemical shift dispersion afforded by 2D NMR and high temporal resolution of 1H NMR in the same experiment. However, interleaving conventional 1D and 2D experiments leaves "time gaps" in each data set. This work presents the Signal And TIme Resolved information Experiment (SATIRE), an NMR supersequence that uses the leftover 1H magnetization (98.9%) not used in the HSQC to collect 1H NMR data within the same scan, using a single receiver. The approach is designed for those interested in monitoring complex systems that have a low signal-to-noise ratio (SNR). For example, if a researcher monitored living organisms for 24 h by 1H NMR alone there may not be enough spectral dispersion for assignment. The low SNR would necessitate a long HSQC, potentially doubling the required NMR time. Conversely, with SATIRE the HSQC would be collected essentially for "free" at the same time as the 1H data are collected. In SATIRE, the signal-to-noise ratios of the HSQC and 1H are identical to the standalone experiments and are designed to permit relative quantitation in the 1H data. Because the data are collected as a pseudo-3D experiment, it is also possible to extract individual shorter time scale HSQC or 1H NMR spectra around time points of interest, SNR permitting. The approach is introduced and then demonstrated on benchtop NMR to monitor sucrose hydrolysis and finally at high field (500 MHz) to follow anoxic stress in vivo. In summary, SATIRE allows simultaneous acquisition of 1D and 2D NMR without compromising either data set and supports complex process monitoring in any application at natural abundance.

Nano liquid chromatography-mass spectrometry has come to be a key enabling technology in shotgun proteomics due to the combination of exceptional separation power, sensitivity, and comprehensiveness. However, the know-how of setting up proteomics methods to deliver robust, reliable, and meaningful results to large-scale life science experiments has remained somewhat ambiguous. This protocol outlines guidance for establishing nano-LC-MS/MS workflows focusing on comprehensive and untargeted deep proteome profiling, using state-of-the-art column technology and mass spectrometry. Employing a second-generation micropillar-array column, a trade-off is demonstrated between analysis time and chromatographic resolving power, which in turn impacts peptide and protein identification scores from a commercial HeLa reference standard. Furthermore, a straightforward workflow to develop a data-independent acquisition (DIA)-parallel accumulation-serial fragmentation (PASEF) analytical method is proposed, with a special focus on the optimization of the ESI source settings. Besides the method development, the study discusses the use of segmented gradients, and an MS-compatible surfactant in the sample diluent is also explored. Finally, the robustness of the developed method is demonstrated through consistently identifying 7558 protein groups (CV=0.3%) as maintaining high repeatability peptide retention times (mean CV=0.2%) and system pressure (CV=0.4%) over 21 consecutive analyses.

Growing global concerns in the energy sector have heightened the focus on developing solutions for more sustainable, cost-efficient, and environmentally friendly energy storage technologies. As advanced energy storage solutions, various battery systems have garnered enormous level of research interest. However, optimizing batteries to meet the demands of rapid charging, enhanced energy density, and longevity is challenging and requires full understanding of their electrochemical properties.Magnetic Resonance techniques, particularly Nuclear Magnetic Resonance (NMR) and Electron Paramagnetic Resonance (EPR), provide insights into the deep understanding of battery chemistry. NMR and EPR studies aim to monitor and understand the local structure of electrodes and electrolyte materials, as well as dynamics and interactions of battery components under operating conditions. Even minor changes in material composition can significantly affect local structures, influencing further material properties and their applicability for electrochemical processes. Magnetic resonance techniques provide unique insights into structural and dynamic changes, enabling a deeper understanding of how these variations influence battery properties.This presentation will provide an overview of the applications of NMR and EPR techniques in battery research, such as charge storage mechanisms of electrodes, electrolytes composition, solid electrolyte interphases, dendrite formation and more. We will highlight how Magnetic Resonance contributes to the development of safer, more efficient, and longer-lasting batteries in both academic and industrial battery R&D settings.

ABSTRACTChaperone-mediated autophagy (CMA) is a selective form of lysosomal protein degradation essential for cellular proteostasis. CMA is activated during cellular stress, such as starvation, and involves the chaperone protein HSC70 (HSPA8) recognizing substrates containing KFERQ-like motifs. However, the regulatory mechanisms governing CMA activation remain poorly understood. Here, we demonstrate that the NAD+ -dependent deacetylase SIRT2 promotes CMA activation by deacetylating HSC70 at lysine 557 (K557). Our findings reveal that SIRT2 activity is upregulated during starvation, enhancing its interaction with HSC70 and facilitating the deacetylation of K557. Deacetylation of HSC70 at K557 increases its binding affinity to CMA substrates, thereby promoting their lysosomal degradation. Mutation of K557 to a deacetylation-mimetic arginine (K557R) enhances CMA activity under both nutrient-rich and starvation conditions, while the acetylation-mimetic glutamine mutant (K557Q) impairs substrate binding and CMA activation. Furthermore, the inhibition or knockdown of SIRT2 reduces CMA activity, which is rescued by HSC70 K557R expression. These findings identify SIRT2-mediated deacetylation of HSC70 as a regulatory mechanism for CMA activation during nutrient deprivation and highlight the role of protein lysine acetylation in proteostasis. This study provides insights into the interplay between SIRT2, HSC70, and CMA, with potential implications for diseases linked to proteostasis dysregulation, including neurodegenerative disorders and cancer.