Slow Cycling Research Articles

Apolipoprotein E is a novel marker for chondrocytes in the growth plate resting zone.

The resting zone (RZ) in mammalian growth plates is critical for maintaining and regulating chondrocyte turnover during longitudinal bone growth as a control tower and stem cell reservoir. Although recent lineage tracing studies have identified several markers for stem cells in the RZ, these markers only partially label chondrocytes in the RZ, suggesting that the resting chondrocytes (RCs) are a heterogeneous population with different types of stem cells. Since a comprehensive marker for RCs is still lacking, the RZ is generally determined based on ambiguous histological criteria, such as small and round chondrocytes without columnar formation, which may lead to inconsistencies among researchers. Therefore, in this study, we used single-cell RNA sequencing (scRNAseq) of growth plate chondrocytes followed by validation by fluorescence in situ hybridization (FISH) to precisely annotate cell clusters in scRNAseq and search for a marker of RCs. The scRNAseq analysis revealed that apolipoprotein E (Apoe) was the top-hit gene, which was ubiquitously expressed in the RC cluster. FISH confirmed that Apoe was exclusively localized to the histologically defined RZ. In newly generated Apoe-mCherry knock-in mice, we further confirmed that mCherry expression mirrored the distribution of Apoe-expressing chondrocytes in the RZ particularly after the formation of the secondary ossification center. These mCherry+ RCs were slow cycling in vivo and exhibited stem cell properties both in vitro and in vivo. Moreover, APOE was detected in human growth plate RCs. These findings suggest that Apoe is a novel pan-RC marker in both mouse and human growth plates.

Ultrafast and stable molten salt aluminum organic batteries

Aluminum-organic batteries (AIBs) have gained significant popularity for large-scale energy storage due to their abundance of aluminum reserves, cost-effectiveness, and environmental friendliness. However, the current aluminum-organic batteries primarily relied on ionic liquid electrolytes suffer from slow reaction kinetics and limited cycle life. Herein, we report a novel and efficient aluminum-organic battery that addresses these limitations by utilizing a molten salt electrolyte and designing a strongly interacting organic cathode. By enhancing π-π stacking interactions, we induced a transition in commercial PTCDA (Perylene-3,4,9,10-tetracarboxylic dianhydride) molecules from the β-phase to the highly interactive α-phase, known as PA450. This transformation not only stabilizes the structure of the PA450 electrode, preventing dissolution in the molten salt electrolyte, but also significantly improves electron conductivity. The Al||PA450 molten salt battery demonstrates exceptional electrochemical performance, exhibiting a high reversible capacity of 135 mAh g–1 and outstanding cyclability for up to 2000 cycles at 10 A g−1. Additionally, the structural rearrangement and ion transport properties induced by the co-intercalation of Al3+ and AlCl2+ were studied are investigated. This work provides deep insights into the unique characteristics of organic materials for ultrafast energy storage in molten salt electrolytes.

How to design optimal brain stimulation to modulate phase-amplitude coupling?

Objective. Phase-amplitude coupling (PAC), the coupling of the amplitude of a faster brain rhythm to the phase of a slower brain rhythm, plays a significant role in brain activity and has been implicated in various neurological disorders. For example, in Parkinson’s disease, PAC between the beta (13–30 Hz) and gamma (30–100 Hz) rhythms in the motor cortex is exaggerated, while in Alzheimer’s disease, PAC between the theta (4–8 Hz) and gamma rhythms is diminished. Modulating PAC (i.e. reducing or enhancing PAC) using brain stimulation could therefore open new therapeutic avenues. However, while it has been previously reported that phase-locked stimulation can increase PAC, it is unclear what the optimal stimulation strategy to modulate PAC might be. Here, we provide a theoretical framework to narrow down the experimental optimisation of stimulation aimed at modulating PAC, which would otherwise rely on trial and error. Approach. We make analytical predictions using a Stuart–Landau model, and confirm these predictions in a more realistic model of coupled neural populations. Main results. Our framework specifies the critical Fourier coefficients of the stimulation waveform which should be tuned to optimally modulate PAC. Depending on the characteristics of the amplitude response curve of the fast population, these components may include the slow frequency, the fast frequency, combinations of these, as well as their harmonics. We also show that the optimal balance of energy between these Fourier components depends on the relative strength of the endogenous slow and fast rhythms, and that the alignment of fast components with the fast rhythm should change throughout the slow cycle. Furthermore, we identify the conditions requiring to phase-lock stimulation to the fast and/or slow rhythms. Significance. Together, our theoretical framework lays the foundation for guiding the development of innovative and more effective brain stimulation aimed at modulating PAC for therapeutic benefit.

Comparative Analysis Of Cycle-To-Cycle Variabilities And Combustion Development In An Optical Spark-Ignition Engine Fueled By Pure Hydrogen And Propane: Insights From Chemiluminescence and PI

Hydrogen, derived from renewable sources and devoid of carbon emissions, is a pivotal energy carrier for the future. Representing a viable substitute for fossil fuels in internal combustion engines (ICEs), contemporary studies advocate using very-lean and ultra-lean hydrogen-air mixtures, with a fuel-air equivalence ratio below 0.5, as a potent strategy to reduce NOx emissions in hydrogen-fueled ICEs. An experimental setup with an optical spark-ignition engine was devised to investigate the primary factors influencing cycle-to-cycle variations in H2ICEs by optimizing mixture homogeneity and focusing the study on flame interaction with in-cylinder aerodynamics. Simultaneous in-cylinder pressure, chemiluminescence, and PIV analyses were performed to characterize and compare the behaviors of ultra-lean hydrogen-air and propane-air flames, assessing flame development and cyclic variation features. Results indicate that the flame development of hydrogen and propane is significantly different. Propane exhibited increased in-cylinder pressure trace variability at lean and rich flammability limits with a high flame development difference between fast and slow cycles. In contrast, hydrogen-air mixtures under various equivalence ratios (from very-lean to ultra-lean) presented much more stable in-cylinder pressures. Furthermore, fast propane flames were mainly advected to the cylinder's symmetrical axes, while fast hydrogen flames started further away from the spark plug. Horizontal and vertical PIV measurements showed a single structure flow field over the studied conditions with mainly intensity variations over the measured cycles. Fast flames were associated with more intense tumble motion. The differences between fast and slow flames for hydrogen are attributed to the early flame development. However, after initial differences in flame development over several crank angles, there are similarities in all cycles, suggesting that the low variability in in-cylinder pressures is mostly due to the robustness of hydrogen flame ignition and its independence from in-cylinder flow small variations at the early development phase.

O-307 Decrease of human sperm hypercondensed chromatin volume after slow freezing revealed by three-dimensional (3D) nuclear morphometry

Abstract Study question Are the 3D morphometry parameters of the human sperm nucleus affected by slow freezing? Summary answer The slow freezing-thawing cycle induces significant alterations on hypercondensed nucleus zone by reducing its volume and splits it into several small zones. What is known already Sperm cryopreservation is an essential technique for infertility care, but it seems to highly affect the genetic and epigenetic integrity of sperm nuclei with poorly identified mechanisms. Moreover, it is known that sperm nuclear alterations may be involved in pregnancy failures. Therefore, sperm nuclear biomarkers are an essential issue to assess the outcome of ART procedures when frozen sperm is used. Thereby, 3D visualization of chromatin is a good way to understand modifications in the organization and composition of sperm chromatin after a slow freezing procedure. Study design, size, duration Ten semen of patients were analyzed (Germetheque biobank) after informed consent. Standard semen parameters, DNA fragmentation, chromatin decondensation, sperm telomere length (STL) and 3D nuclear morphometry were performed before and after slow freezing. Morphometric measurements were performed from a digital representation of the sperm nucleus in 3 dimensions that allow us to measure the volume, flatness and elongation of each sperm nucleus and characterize hypercondensed zones, with live/dead spermatozoa distinction. Participants/materials, setting, methods After addition of cryoprotective medium v/v (Cryosperm®), sperm was frozen with a programmable freezer (NanoDigitcool®). Standards sperm parameters were measured in accordance with WHO guidelines 2021, DNA fragmentation by flow cytometry TUNEL assay, chromatin decondensation by epifluorescent microscopy with Chromomycin A3 and STL using qPCR. 3D nuclear representation was performed by a z-axis image acquisition series of DAPI/PI stained slides with optigrid epifluorescent microscopy. Analyses were carried out using the NucleusJ2.0 software with NODeJ plugin. Main results and the role of chance Freezing induces an alteration of sperm vitality (p < 0.0001) and progressive and total motility (p < 0.001 and p < 0.0001 respectively) and an increase in the percentage of sperm with DNA fragmentation (18.30% fresh vs 31.6% frozen, p < 0.05). We did not observe a modification of the median volume of the sperm nucleus before and after freezing (37.15 [IC = 36.5;38.14] and 37.10 [33.27;37.78] µm3 respectively), as the same for flatness and elongation (2.48 vs. 2.50 a.u. and 2.04 vs. 2.04 a.u. respectively). Nevertheless, five samples showed significant alterations of nucleus volume (p < 0.0001), due to diminution of flatness and more or less elongated. A significant decrease of the total volume of the hypercondensed chromatin zone was observed after freezing (0.94 [0.84;0.99] before vs. 0.61 [0.53;0.69] µm3 after freezing-thawing, p < 0.0001), with an effect size of -1.48 [-2.10; -0.86]. Decrease of total volume of hypercondensed region was measured homogeny in all the samples tested, particularly on live (1.00 vs. 0.68 µm3 respectively, p < 0.0001) compared with dead spermatozoa (0.72 vs. 0.57 µm3, p < 0.05). We showed also a significant increase of the number of hypercondensed zones in each nucleus (p < 0.001), reflecting a split into several parts of the hypercondensed chromatin. Limitations, reasons for caution A 3D-FISH analysis would allow to characterize the hypercondensed chromatin portion, probably as heterochromatin and which we suspect to be the nucleus ring. Moreover, a chromosomes specific marking would allow for the identification of whether whole chromosomes or specific parts of chromosomes (centromeres or telomeres) are affected by this decompaction. Wider implications of the findings This study is the first to make exacts measurements of human sperm nucleus because of a non-decondensing method. We demonstrate that slow freezing affects hypercondensed chromatin probably through a decondensation and modification of the nucleus-specific organization. This new assessment will allow a better proficiency of men fertility preservation techniques. Trial registration number NCT04715828

Rapid response to pressure variations in water distribution networks through machine learning-enhanced data acquisition

ABSTRACT This study investigates rapid dynamic pressure variations in water distribution networks due to critical incidents such as pipe bursts and valve operations. We developed and implemented a machine learning (ML)-based methodology that surpasses traditional slow cycles of pressure data acquisition, facilitating the efficient capture of transient phenomena. Employing the Orion ML library, which features advanced algorithms including long short-term memory dynamic threshold, autoencoder with regression, and time series anomaly detection using generative adversarial networks, we engineered a system that dynamically adjusts data acquisition frequencies to enhance the detection and analysis of anomalies indicative of system failures. The system's performance was extensively tested using a pilot-scale water distribution network across diverse operational conditions, yielding significant enhancements in detecting leaks, blockages, and other anomalies. The effectiveness of this approach was further confirmed in real-world settings, demonstrating its operational feasibility and potential for integration into existing water distribution infrastructures. By optimizing data acquisition based on learned data patterns and detected anomalies, our approach introduces a novel solution to the conventionally resource-intensive practice of high-frequency monitoring. This study underscores the critical role of advanced ML techniques in water network management and explores future possibilities for adaptive monitoring systems across various infrastructural applications.

Integrating scRNA-seq and bulk RNA-seq to explore the differentiation mechanism of human nail stem cells mediated by onychofibroblasts.

Introduction: Nail stem cell (NSC) differentiation plays a vital role in maintaining nail homeostasis and facilitating digit regeneration. Recently, onychofibroblasts (OFs), specialized mesenchymal cells beneath the nail matrix, have emerged as potential regulators of NSC differentiation. However, limited understanding of OFs' cellular properties and transcriptomic profiles hinders our comprehension of their role. This study aims to characterize human OFs and investigate their involvement in NSC differentiation. Methods: Human OFs were isolated and characterized for their mesenchymal stem cell (MSC)-like phenotype through flow cytometry and multilineage differentiation assays. Bulk RNA-seq analysis was conducted on three samples of OFs and control fibroblasts from human nail units to delineate their molecular features. Integrated analysis with scRNA-seq data was performed to identify key signaling pathways involved in OF-induced NSC differentiation. Co-culture experiments, siRNA transfection, RT-qPCR, and immunocytochemistry were employed to investigate the effect of OF-derived soluble proteins on NSC differentiation. Drug treatments, RT-qPCR, western blotting, and immunocytochemistry were used to verify the regulation of candidate signaling pathways on NSC differentiation in vitro. Results: Human OFs exhibited slow cell cycle kinetics, expressed typical MSC markers, and demonstrated multilineage differentiation potential. Bulk RNA-seq analysis revealed differential gene expression in OFs compared to control fibroblasts, highlighting their role in coordinating nail development. Integrated analysis identified BMP4 as a pivotal signal for OFs to participate in NSC differentiation through mesenchymal-epithelial interactions, with the TGF-beta pathway possibly mediating this signal. OFs synthesized and secreted more BMP4 than control fibroblasts, and BMP4 derived from OFs induced NSC differentiation in a co-culture model. Recombinant human BMP4 activated the TGF-beta pathway in NSCs, leading to cell differentiation, while the BMP type I receptor inhibitor LDN193189 attenuated this effect. Discussion: This study characterizes the cellular and molecular features of human OFs, demonstrating their ability to regulate NSC differentiation via the TGF-beta signaling pathway. These findings establish a connection between the dermal microenvironment and NSC differentiation, suggesting the potential of OFs, in conjunction with NSCs, for developing novel therapies targeting nail and digit defects, even severe limb amputation.

Abstract PO4-28-01: Molecular profiling revealed activation of cytokine signaling and suppression of lipid metabolism as the hallmarks of doxorubicin-resistant TNBC

Abstract Background: Chemotherapeutic resistance observed in triple-negative breast cancer (TNBC) poses a substantial clinical hurdle, emphasizing the imperative to enhance our comprehension of the underlying mechanisms with the aim of potentially mitigating or reversing this phenomenon. Methods: Doxorubicin-resistant (DoxR) TNBC models (MDA-MB-231 and BT-549) were established through continuous exposure to increasing concentrations of doxorubicin. RNA-Seq was conducted using the Illumina platform, while bioinformatics analyses were performed using CLC genomics workbench 20.2 and iDEP.91. Colony forming unit (CFU), flow cytometry and fluorescent microscopy were used to assess cell proliferation, cell cycle distribution, and cell death, respectively. Western blotting was used to confirm the expression and phosphorylation of protein targets. Ingenuity pathway analysis (IPA) and STRING database v 11.5 were used for network and pathway analyses, while Kaplan-Meier Plotter was used for survival analyses. Results: Herein, we provide a comprehensive functional and transcriptomic characterization of DoxR TNBC models, revealing multiple affected signaling networks and functional categories. The defense and immune response, response to stress, response to cytokine and external stimulus and cytokine-mediated signaling pathways were among the top activated, while suppression of cholesterol, sterol and lipid biosynthesis and metabolism were the hallmarks of DoxR cells. Upstream regulator analysis revealed IL-1B cytokine network activation in DoxR cells. Concordantly, disease and function analysis on the downregulated genes in DoxR TNBC cells revealed the most pronounced enrichment in pathways associated with cell invasion, colony formation, as well as lipid and carbohydrate metabolism. A number of upregulated genes were validated by qRT-PCR, which correlated with unfavorable overall survival (OS). Functionally, DoxR cells exhibited remarkable suppression of cell proliferation, as evidenced by diminished CFU potential, sphere formation and growth under 3D organoid culture, and cell migration. Although DoxR TNBC cells exhibited a slow cell cycling under normal culture conditions, wild-type TNBC cells exhibited remarkable arrest in the G2M phase when exposed to doxorubicin, which was less evident in DoxR TNBC models, suggesting DoxR cells undergo a quiescent state in response to chemotherapeutic challenge. In agreement with those data, increased phosphorylation of CHK2 and p53 proteins was remarkable in DoxR cells, thus promoting cellular dormancy. Conclusion: Our data revealed cellular dormancy and suppression of lipid metabolism as the hallmark associated with doxorubicin-resistant TNBC cells. On the other hand, cytokine signaling and stress response were the most enriched functional categories in DoxR TNBC cells. Therapeutic targeting to reverse the quiescent state or suppression of cytokine signaling networks could enhance the efficacy of doxorubicin-based therapeutics. Citation Format: Vishnubalaji Radhakrishnan, Nehad Alajez. Molecular profiling revealed activation of cytokine signaling and suppression of lipid metabolism as the hallmarks of doxorubicin-resistant TNBC [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-28-01.

Abstract LT05: METTL8-MEDIATED EPITRANSCRIPTOMIC REGULATION OF GBM CYCLING STATE IS DEPENDENT ON RTK/PI3K/AKT SIGNALING

Abstract Glioblastoma (GBM) is a lethal brain tumor with limited treatment options. Treatment failures are often attributed to the existence of glioma stem cells (GSCs), which confer intratumoral heterogeneity and therapy resistance. Contrary to common belief that cancer cells employ aerobic glycolysis (Warburg effect) to sustain their metabolic requirements, GSCs rely on oxidative phosphorylation (OXPHOS) for energy production. As a result, GSCs are uniquely sensitive to OXPHOS inhibitors (Gboxin), as well as mitochondrial translation inhibitors which disrupt OXPHOS. METTL8 is a newly discovered mitochondrial methyltransferase which catalyzes m3C32 modifications in mitochondrial tRNAsThr/Ser(UCN) (mt-tRNAs). This post-transcriptional modification ensures the optimal structure of mt-tRNAs to facilitate efficient translation of mitochondrial proteins involved in electron transport chain (ETC). Although the epitranscriptomic functions of METTL8 have been well characterized, its roles in cancers (i.e. glioblastoma) remain largely unexplored. In this study, we showed that METTL8 is overexpressed in GBM and high METTL8 expression portends poor patient prognosis. In GSC, METTL8 is under the transcriptional regulation of HIF-1α, and the chromatin accessibility at METTL8 promoter is facilitated by the deposition of histone variant H2AZ. METTL8 depletion downregulated m3C32 modifications in mt-tRNAsThr/Ser(UCN) in GSC, leading to impaired mitochondrial translation and OXPHOS defects. Phenotypically, METTL8 knockdown (KD) compromised GSCs’ self-renewal, proliferation, invasiveness and tumorigenicity. Interestingly, METTL8-KD GSC acquired a slow cycling state which is linked to reduced receptor tyrosine kinase (RTK) signaling. Consistently, METTL8-KD GSC showed lower Akt phosphorylation and exhibited increased sensitivity to mTOR/Akt inhibitors. Finally, our mechanistic findings led to the identification of a novel HIF1-α/Akt inhibitor drug combination which faithfully recapitulates the phenotypes of METTL8-KD GSCs. In conclusion, our findings provide the first in-depth study of METTL8 roles in GBM pathogenesis, and highlights a novel drug combination which could potentially be used to treat this dreadful disease. Citation Format: Bernice Woon Li Lee, You Heng Chuah, Jeehyun Yoon, Oleg V. Grinchuk, Jia Feng, Yajing Liang, Jayshree L Hirpara, Ya Ting Shen, Loo Chien Wang, Yan Ting Lim, Tianyun Zhao, Radoslaw M Sobota, Tan Boon Toh, Shazib Pervaiz, Zhewang Lin, Derrick Sek Tong Ong. METTL8-MEDIATED EPITRANSCRIPTOMIC REGULATION OF GBM CYCLING STATE IS DEPENDENT ON RTK/PI3K/AKT SIGNALING [abstract]. In: Proceedings of Frontiers in Cancer Science; 2023 Nov 6-8; Singapore. Philadelphia (PA): AACR; Cancer Res 2024;84(8_Suppl):Abstract nr LT05.

Superoxide radical-induced rapid valence cycling of Mn(II/III/IV) for efficient degradation of organic pollutants

Transition metal oxides are one of the commonly used catalysts for ozone oxidation, and the slow redox cycling of different valence metal components inside the catalysts currently becomes a key bottleneck limiting their catalytic activity. In this work, the superoxide radicals generated at the N/C sites were used to reduce the neighboring high-valence metal components to low-valence states to construct a new mechanism for the rapid cyclic transition of Mn (II/III/IV) valence states. What's more, N/C sites enhanced the ozone adsorption and conversion ability, synergized with the adjacent MnO2 sites to generate interfacial hydroxyl radicals and superoxide radicals, and effectively inhibited the quenching effect of inorganic salt ions (e.g., Cl-, SO42-) on the free radicals. For the actual printing and dyeing wastewater, the COD (chemical oxygen demand) removal rate was as high as 70 % in 60 min and remained stable in 10 cycles with almost no manganese ions leaching. In addition, characterization such as EPR combined with DFT calculations showed that the dual-site structure significantly increased the oxygen vacancy content and Lewis acidity, and the synergistic interaction between graphite N and MnO2 was the most favorable for ozone conversion with the lowest reaction energy barriers.

Abstract LB332: Spatial transcriptomic and proteomic analysis of the treatment resistant slow cycling micro environment in GBM

Abstract Within the Glioblastoma (GBM) microenvironment, there are micro-niches originating from independent lineages of cancer stem cells with distinct metabolic requirements that are characterized by Fast cycling cells (FCCs) and Slow cycling cells (SCC). The FCCs employ aerobic glycolysis for energy, while the treatment-resistant SCCs primarily rely on lipid metabolism. Our study allows for the molecular and spatial decoding of the heterogeneity within the GBM microenvironment, with a specific focus on decoding the dichotomy of the immune infiltrates and studying the interactions between the SCCs and the immune compartment in human GBM patients. Our research reveals a new paradigm of interaction within the tumor microenvironment (TME) of glioblastoma (GBM) patients, in which SCCs are instrumental in altering the tumor milieu to favor immune suppression. These SCCs actively recruit and harness sub-population of immune cells of myeloid origin that promote tumor growth and simultaneously provide metabolic aid to these treatment-resistant cells. Our research maps out the intricate metabolic dialogues within GBM and evaluates the potential therapeutic effect of disrupting this interaction to open up new treatment avenues. The findings based on our spatial transcriptomic and proteomic study on brain biopsies from human patients provide a deeper understanding of the critical links among the tumor's local environment, cellular metabolism, anti-tumor immunity, and potential treatment vulnerabilities. In summary, our investigation offers novel insights into how metabolic exchanges contribute to the evolution of GBM leading to the development of drug and immune resistant GBM. Our data is suggestive that therapies aimed at these metabolic interactions, especially those between SCCs and metabolically supporting immunosuppressive myeloid originating immune cells, hold potential as effective therapeutic strategy for GBM. Citation Format: Avirup Chakraborty, Diana Feier, Aryeh Silver, Changlin Yang, Miruna Anica, Avinash Pittu, Olusegun Sobanjo, Ethan Hodge, Mia Engelbart, Christina Von Roemeling, Maryam Rahman, Matthew Sarkisian, Jianping Huang, Jeffrey Harrison, Duane Mitchell, Loic Deleyrolle. Spatial transcriptomic and proteomic analysis of the treatment resistant slow cycling micro environment in GBM [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB332.

Abstract LB293: O-GlcNAc transferase drives the proliferative state in adaptive drug tolerance

Abstract Drivers of the drug tolerant proliferative persister (DTPP) state have not been well investigated. Histone H3 lysine-4 trimethylation (H3K4me3), an active histone mark, might enable slow cycling drug tolerant persisters (DTP) to regain proliferative capacity. This study aimed to determine H3K4me3 transcriptionally active sites identifying a key regulator of DTPPs in melanoma. Deploying a model of adaptive cancer drug tolerance H3K4me3 ChIP-seq data of DTPPs guided identification of top transcription factor binding motifs. These suggested involvement of O-linked N-acetylglucosamine transferase (OGT), which was confirmed by metabolomics analysis and biochemical assays. OGT impact on DTPPs and adaptive resistance was explored in vitro and in vivo. H3K4me3 remodeling was widespread in CPG island regions and DNA binding motifs associated with O-GlcNAc marked chromatin. Accordingly, we observed an upregulation of OGT, O-GlcNAc and its binding partner TET1 in chronically treated cancer cells. Inhibition of OGT led to loss of H3K4me3 and downregulation of genes contributing to drug resistance. Genetic ablation of OGT prevented acquired drug resistance in vivo. Our findings uncover a fundamental mechanism of adaptive drug resistance that governs cancer cell reprogramming towards acquired drug resistance, a process that can be exploited to improve response duration and patient outcomes. Citation Format: Dinoop Ravindran Menon, Heinz Hammerlindl, Gregory Gimenez, Elmar Zuegner, Christoph Magens, Michael Eccles, Mayumi Fujita, Helmut Schaider. O-GlcNAc transferase drives the proliferative state in adaptive drug tolerance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB293.

Field and laboratory life cycle studies of Malenka depressa (Banks, 1898) (Plecoptera: Nemouridae)

During 1982–1985, laboratory and field studies of 125 eggs, 8055 nymphs and 1426 adults indicated a univoltine slow life cycle for Malenka depressa (Banks, 1898) in a first-order perennial stream in the Mediterranean climate of the San Francisco Bay area. Nymphal growth and adult emergence were continuous throughout the year, further enhanced by a thick, protective tree canopy shading of the canyon that provided an insulated stabilised thermal tube of cool air, ground and stream water temperatures. Leaf senescence of the overhead California bay trees was a perennial source of allochthonous food for the immature stages. Maximum assimilation efficiency for middle to late-nymphal growth stages fed on bay leaves was 42% at 10 °C.

Estimation of Sediment Transport Parameters From Measured Suspended Concentration Time Series Under Waves and Currents With a New Conceptual Model

AbstractIn‐situ observations of hydrodynamics and suspended sediment concentrations (SSCs) were conducted on an abandoned lobe in the northern part of the modern Yellow River Delta, China. The SSC record at the site is found to be the superposition of a general trend (fast increase and slow decrease cycle) caused by storm waves (SubSSC1) and relatively smaller fluctuations caused by tidal currents (SubSSC2). Physically, this indicates that storm waves eroded the bottom sediments while tidal currents then re‐suspended and advected the suspended sediments in the study area. To further obtain the suspended sediment transport parameters, first, SubSSC1 is modeled with significant wave height which incorporates a “memory curve” to consider the remaining impacts of historical waves. It is detected that waves in the past 75 hr still influence the present SSC which is reasonable because 75 hr is roughly the typical duration of a normal storm. Second, SubSSC2 is modeled with tidal excursion and trigonometric functions with measured periodicities. Finally, some sediment transport parameters, for example, the background SSC, the horizontal SSC gradient, the tidal constituents that advect it, and their relative time lags are optimized from the best fits of the measured and modeled SSC time series. The proposed framework for model construction and parameter optimization can be extended to other sea areas for inferring sediment transport parameters from field SSC time series at a specific station.

Abstract 3083: Unveiling the role of chemotherapy-resistant, slow-cycling cancer cells in driving cancer cachexia

Abstract Cancer cachexia, a systemic disease marked by substantial, involuntary weight loss, predominantly affecting muscle and adipose tissue, manifests in patients with advanced-stage cancer. The emergence of cancer cachexia poses significant challenges to cancer treatment, diminishing the efficacy of anticancer medications, and the resultant muscle loss adversely impacts the daily lives of patients, diminishing the overall quality of life. Notably, cancer cachexia is implicated in 30-40% of cancer-related deaths, underscoring the pivotal need to comprehend its underlying principles and propose apt interventions to enhance both survival rates and the well-being of cancer patients. Based on our previous findings, cancer cells exhibiting a slow cell cycle, resistant to widely employed clinical chemotherapy agents, exhibit the formation of proinflammatory tumor microenvironment. The escalation of systemic inflammatory cytokines has long been suggested as a potential mechanism contributing to cancer cachexia. Building upon these insights, we hypothesized that slow-cycling cancer cells (SCCs) featuring resistance to chemotherapy are pivotal contributors to cancer cachexia. We investigated the impact of drug-resistant SCCs on the loss of differentiated myotubes and lipid droplets in vitro and muscle and fat loss in in vivo conditions. Interestingly, these cells induced more severe muscle and fat losses. Upon scrutinizing this cell population, we found the overexpression of a particular membrane protein, Protein A. The increase of this Protein A enhanced the release of exosomes from the cell through a novel mechanism distinct from its classical functions, consequently amplifying the release of inflammatory substances originating from cancer cells. In summary, our study elucidates that chemotherapy-resistant cancer cells, characterized by a slow cell cycle, intensify the secretion of inflammatory substances in a manner contingent upon the overexpression of a membrane protein A, which leads to the depletion of muscle and fat located in the distal peripheral regions. Citation Format: Jaebeom Cho, Ho-Young Lee. Unveiling the role of chemotherapy-resistant, slow-cycling cancer cells in driving cancer cachexia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3083.

Health benefits of interval walking training.

Interval walking training (IWT) is a free-living training intervention involving alternating fast and slow walking cycles. IWT is efficacious in improving physical fitness and muscle strength, and reducing factors associated with lifestyle-related diseases. In individuals with type 2 diabetes, IWT improves glycemic control directly through enhanced glucose effectiveness, challenging conventional views on mechanisms behind training-induced improvements in glycemic control. Whereas adherence to IWT in short-term studies is high, ensuring long-term adherence remains a challenge, particularly in populations with chronic diseases and/or overweight/obesity. Long-term studies in real-world settings are imperative to ascertain the widespread effectiveness of IWT and elucidate its impact on hard endpoints.

Increasing Activity of Trimetallic Oxygen Reduction PtNiMo/C Catalysts Through Initial Conditioning

AbstractThis study investigates the importance of preconditioning in Pt‐alloy catalysts for oxygen–reduction reactions. Previous research indicated that slower scanning rates during preconditioning initially boost activity, but this is followed by a rapid decline. The study reveals the required number of cycles to achieve the first constant steady state activity in PtNiMo/C catalysts when using slower scanning rates during preconditioning. It also highlights the resulting activity differences. Remarkably, a catalyst preconditioned with 150 slow cycles showed an activity of approximately 1.25 mA cm−2 at 0.90 VRHE, significantly higher than one preconditioned at a fast rate (0.82 mA cm−2). Both ex–situ and in–situ analyses revealed that Pt, along with Ni, was leached during pretreatment. At slower scan rates of 20 mV s−1, the dissolved Pt redeposited as highly active, small‐sized clusters or single atoms. Fast scan rates of of 500 mV s−1, in contrast, resulted in fewer such clusters. Accelerated stress tests up to 1.10 VRHE confirmed the high stability of these clusters, demonstrating a substantial activity increase even after 24,000 cycles.

Moisture and temperature effects on the radiocarbon signature of respired carbon dioxide to assess stability of soil carbon in the Tibetan Plateau

Abstract. Microbial release of CO2 from soils to the atmosphere reflects how environmental conditions affect the stability of soil organic matter (SOM), especially in massive organic-rich ecosystems like the peatlands and grasslands of the Qinghai–Tibetan Plateau (QTP). Radiocarbon (14C) is an important tracer of the global carbon cycle and can be used to understand SOM dynamics through the estimation of time lags between carbon fixation and respiration, often assessed with metrics such as age and transit time. In this study, we incubated peatland and grassland soils at four temperature (5, 10, 15 and 20 °C) and two water-filled pore space (WFPS) levels (60 % and 95 %) and measured the 14C signature of bulk soil and heterotrophic respired CO2. We compared the relation between the Δ14C of the bulk soil and the Δ14CO2 of respired carbon as a function of temperature and WFPS for the two soils. To better interpret our results, we used a mathematical model to analyse how the calculated number of pools, decomposition rates of carbon (k), transfer (α) and partitioning (γ) coefficients affect the Δ14C bulk and Δ14CO2 relation, with their respective mean age and mean transit time. From our incubations, we found that 14C values in bulk and CO2 from peatland were significantly more depleted (old) than from grassland soil. Our results showed that changes in temperature did not affect the Δ14C values of heterotrophic respired CO2 in either soil. However, changes in WFPS had a small effect on the 14CO2 in grassland soils and a significant influence in peatland soils, where higher WFPS levels led to more depleted Δ14CO2. In our models, the correspondence between Δ14C, age and transit time highly depended on the internal dynamics of the soil (k, α, γ and number of pools) as well as on model structure. We observed large differences between slow and fast cycling systems, where low values of decomposition rates modified the Δ14C values in a non-linear pattern due to the incorporation of modern carbon (14C bomb) in the soil. We concluded that the stability of carbon in the peatland and grassland soils of the QTP depends strongly on the direction of change in moisture and how it affects the rates of SOM decomposition, while temperature regulates the number of fluxes. Current land cover modification (desiccation) in Zoigê peatlands and climate change occurring on the QTP might largely increase CO2 fluxes along with the release of old carbon to the atmosphere potentially shifting carbon sinks into sources.

The logic of carbon substitution: from fossilised life to “cell factories”

This paper examines how researchers in biotechnology reflect on the challenges of turning microbes into what they call “cell factories”. These researchers use the tools of genome editing to harness the biochemistry of single cell organisms, such as bacteria, yeasts and microalgae, and tweak the enzymatic reactions of their metabolism. One research priority is to engineer microbes able to feed on agricultural residues and assemble drop-in compounds to be used in a range of commercial products, from drugs and food additives, to cosmetics, detergents and fuels. To justify financial support for such research, arguments about the need to move away from petroleum as a source of energy and feedstock for chemical synthesis are put forward, underpinned by concerns for climate change, resource renewability and energy security. Drawing on interviews with scientists, we explore what it means for them to make “cell factories” and discuss how they problematise the logic of carbon substitution that orientates their work. Biotechnology is expected to support a shift from one source of carbon, past life gone through slow geological cycles, to a different source of carbon, renewable biomass metabolised by living microbes. As scientists face unhappy cells, recalcitrant plant fibres and unfair competition from fossil-based processes, the promise of carbon substitution tends to be most convincing in the confined space of the lab where faith in biotechnology goes hand in hand with a pragmatic commitment to sustainability. We speculate that the researchers might be failed by the system that biotechnology seeks to (partially) replace, the conditions of which are shaped not around the material constraints of making “cell factories”, but around fossilised life cracked in ever-greater quantities.

Study on Condition Monitoring of Pitch Bearings Based on Stress Measurement

Pitch bearings in wind turbines are crucial components that enable safe blade pitching, optimize electrical power output, and ensure turbine protection. Traditional vibration analysis-based methods used for high-speed bearings are not applicable to monitoring pitch bearings, due to its slow non-integer cycle rotation. To address this issue, a stress-based pitch bearing monitoring method is proposed in this paper. First, finite element analysis is conducted to establish the relationship between the maximum surface stress on the outer race of the pitch bearing and the presence of cracks. This relationship allows the identification of cracks on the outer race and an assessment of their severity based on the value of the maximum surface stress. Second, the outer race of the pitch bearing is divided into several segments, and a singularity detection technique is employed to locate the position of cracks on the outer race based on the stresses measured from the segments. To verify the proposed method, a wind turbine pitch bearing test rig was developed in a laboratory. Experimental results have shown that the proposed method can effectively and accurately identify and locate cracks on the outer race of the bearing, thereby demonstrating its great potential as a reliable approach for monitoring the condition of wind turbine pitch bearings.