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Biology subjects

Wang, F.

Publications and source records attributed to Wang, F..

At least 91 records · Page 5Linked to original sources

Cancer drives atherosclerotic plaque vulnerability by inducing pathological angiogenesis

Cardiovascular disease (CVD) is the leading cause of death for many cancer survivors, a phenomenon traditionally attributed to shared risk factors and cardiotoxic chemotherapies. Here, we hypothesized that cancer may also directly promote atherosclerosis. Propensity-matched analyses confirmed significantly elevated cardiovascular event rates amongst cancer patients, independent of comorbidities such as smoking. Atheroprone mice implanted with colorectal tumors demonstrated accelerated features of plaque vulnerability, driven by pathological angiogenesis and intraplaque hemorrhage. Mechanistically, tumor-secreted TNF- induced the pro-angiogenic factor LRG1 across multiple murine models and human plaques. Therapeutic interventions targeting these pathways, including with FDA-approved cytokine inhibitors or tumor resection, prevented plaque destabilization in mice and reduced coronary revascularization rates in patients. Together, these findings suggest that cancer may causally promote CVD and unveil novel translational strategies for cancer survivors.

pathology↗

A reconfigurable DNA memory architecture for hierarchical data management via programmable phase transitions

DNA has emerged as a promising medium for the post-silicon era of information storage due to its ultrahigh density and longevity. However, current systems are bifurcated, with solid-state systems providing robust cold archival but lacking accessibility, while fluidic molecular computing systems offer dynamic processing but suffer from low density and instability. This mutual exclusivity has hindered the development of hierarchical memory, a standard in modern computing, within molecular storage systems. Here, we bridge this gap by engineering a reconfigurable DNA memory architecture driven by programmable liquid-liquid phase separation (LLPS). Our system leverages sequence-based encoding to achieve an ultrahigh storage density of 7x1010 GB/g, approaching the theoretical limits of DNA accessibility. In its fluidic hot state, DNA droplets enable rapid data loading ([~]83.8% in 5 min) and function as an in-memory editing platform supporting versatile, addressable bit-level operations including selective erasure ([~]65.1%) and high-efficiency rewriting and replacement (>99%) via programmable strand displacement. Importantly, to resolve the stability trade-off, we engineered a programmable phase transition whereby the triggered assembly of a rigid tetrahedral DNA framework (TDF) armor transforms liquid condensates into robust armored droplets. This cold state confers exceptional resistance to enzymatic and physical degradation, projecting multi-millennial data stability. By enabling reversible transitions between a editable, high-density computing mode and a stabilized archival mode, this work establishes the architectural foundation for scalable molecular information storage capable of hierarchical data management.

synthetic biology↗

THIK-1 channel mediates microglial glucose sensing and modulates AgRP neurons

Microglia play essential roles in maintaining energy homeostasis, and their dysfunction contributes to metabolic disease. Although high-fat diet (HFD) exposure induces microglial activation, the underlying mechanisms remain poorly defined. Here, we identified a previously unrecognized role for THIK-1 channel in mediating glucose sensing of microglia in arcuate nucleus of the hypothalamus (ARH), during HFD-induced obesity. Pharmacological inhibition of THIK-1 channel with tetrapentylammonium (TPA) suppresses feeding and attenuates body-weight gain in diet induced obese mice. Mechanistically, inhibition of agouti-related peptide (AgRP) neurons is indispensable for TPA-induced hypophagia. Moreover, THIK-1 inhibition promotes microglial phagocytosis of perineuronal nets (PNNs), leading to reduced AgRP neuronal activity and feeding suppression. Together, these findings establish THIK-1 as a critical glucose sensor in hypothalamic microglia and uncover a microglia-dependent pathway through which overnutrition modulates AgRP neuronal activity via PNN remodeling to regulate energy balance, highlighting THIK-1 as a potential therapeutic target for treatment of diet-induced obesity.

neuroscience↗

The overall and sequence-specific degradation of soil extracellular 16S rRNA genes across China: rates and influential factors

While extracellular DNA persistence substantially influences soil microbiome investigations, its degradation kinetics remain poorly quantified. Here, we developed a primer-labeled DNA approach coupled with microcosm incubation to determine the overall and sequence-specific degradation rates of extracellular DNA amplicon fragments across China. We observed substantial variations in the overall degradation rates of extracellular 16S rRNA gene amplicon fragments among the study sites, with degradation rate constants ranging from 0.05 to 0.16 day-1. The overall degradation rate constants showed significant correlations with soil moisture content, prokaryotic abundance, prokaryotic community profiles, and mean annual precipitation (MAP). The significant influences of moisture content on the overall degradation rates were further verified by a moisture gradient microcosm experiment. The sequence-specific degradation rate constant profiles were additionally correlated with pH, nitrogen content, and mean annual temperature (MAT). Furthermore, propidium monoazide (PMA)-based exclusion of extracellular DNA signals significantly altered soil prokaryotic abundance, richness, and prokaryotic community profiles, and the pool sizes of sequence-specific extracellular 16S rRNA gene amplicon fragments were significantly correlated with their respective degradation rates. This study developed a methodology for determining the overall and sequence-specific degradation rates of extracellular DNA amplicon fragments, highlighting the profound influences of extracellular DNA on soil microbial research and informing the optimization of environmental DNA technologies. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/699460v2_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@b1b3e2org.highwire.dtl.DTLVardef@98f334org.highwire.dtl.DTLVardef@1870650org.highwire.dtl.DTLVardef@1af7105_HPS_FORMAT_FIGEXP M_FIG C_FIG

ecology↗

Development of Integrin α5β1-targeted PET/NIR imaging probes for glioblastoma intraoperative navigation and intracavity targeted radionuclide therapy

Incomplete extent of tumor resection is the major factor of glioblastoma recurrence, leading to its poor prognosis. Developing therapies that enable the minimal tumor cell remnant during surgical resection is a major challenge to combat this aggressive cancer. This study develops a glioblastoma integrin 5{beta}1-selective peptide binder through series strategy screening and labeling it with 68Ga, indocyanine green (ICG), or 177Lu for efficacious multimodal treatment. This strategy combines positron emission tomography (PET) imaging for preoperative diagnosis, intraoperative NIR-II fluorescence-guided surgery for maximal tumor resection, and followed by intracavity targeted radionuclide therapy for elimination of residual tumor cells. We identified GR, with five arginine mutation, exhibiting superior integrin 5{beta}1 binding affinity and brain penetration. [68Ga]GR demonstrated substantial tumor uptake and prolonged retention time in both mouse models and tissues from glioblastoma patients. The guidance of ICG-GR achieved accurately intraoperative tumor imaging and maximal tumor resection in orthotopic mouse glioblastoma models. Notably, combining intracavity administration of [177Lu]GR following ICG-GR guiding resection markedly inhibited tumor growth and reduced possibility of tumor recurrence compared with surgery alone (unguided or ICG-GR-guided) or unguided surgery followed by [177Lu]GR. Therefore, we have reported the integrin 5{beta}1-targeted radionuclide and optical agents for maximal elimination of tumor cells during surgical resection in glioblastoma patients. HIGHLIGHTSO_LIGR is a glioblastoma-specific peptide with both favorable binding affinity superior brain penetration. C_LIO_LIICG-GR is a glioblastoma-targeted NIR probe enabling maximal surgical resection. C_LIO_LIICG-GR-guided surgery with [177Lu]GR-based adjuvant therapy allows robust elimination of tumor cells. C_LI

cancer biology↗

Designing AI-programmable therapeutics with the EDEN family of foundation models

The ability to interpret, modify, and design DNA has driven many of the most significant advances in modern medicine, from diagnostics, biologics, and vaccines to cell and gene therapies. However, the inherent complexity of biological systems means that most modern medicines are still engineered using bespoke, labor-intensive processes. To address the need for a generalisable and programmable approach to therapeutic design, we introduce the EDEN (environmentally-derived evolutionary network) family of metagenomic foundation models, including a 28 billion parameter model trained on 9.7 trillion nucleotide tokens from BaseData1. This dataset, at the time of training, contained more than 10 billion novel genes from over 1 million new species, and is intentionally enriched for environmental and host-associated metagenomes, phage sequences, and mobile genetic elements, enabling the model to learn from diverse and novel cross-species evolutionary mechanisms and apply them to key challenges in human health. EDEN achieves state-of-the-art performance across a series of predictive and generative genomic and protein benchmarks. To demonstrate the models broad applicability across biology, we evaluate EDENs capacity for programmable therapeutic design by challenging a single architecture to design biological novelty across three distinct therapeutic modalities, disease areas and biological scales: (i) large gene insertion, (ii) antibiotic peptide design, and (iii) microbiome design. First, we demonstrate AI-programmable Gene Insertion (aiPGI), in which EDEN designs de novo large serine recombinases (LSRs) capable of inserting large pieces of DNA at desired target sites in the human genome when prompted only on 30 nucleotides of DNA sequence from the desired target site. In low-N experimental validation, EDEN generated multiple active recombinases for all tested disease-associated genomic loci (ATM, DMD, F9, FANCC, GALC, IDS, P4HA1, PHEX, RYR2, USH2A) and 4 potential safe harbor sites in the human genome. EDEN achieves an overall functional hit rate of 63.2% across diverse DNA prompts when prompted on only 30bp of DNA from outside the training data. 50% of EDEN-generated LSRs were active in human cells, achieving therapeutically relevant levels of CAR insertion in primary human T cells. We also show that EDEN can generate active bridge recombinases when prompted on the associated guide RNA alone, with sequence identities to training and public data as low as 65%. These results pave the way for a new generation of cell and gene therapies by opening the door to rapid, programmable and site-specific integration of large genetic payloads without double-strand breaks. This offers an alternative to the safety, efficiency and payload limitations inherent in viral or nuclease-based editing at thousands of currently intractable human therapeutic targets. Second, we use the same model to generate a focused low-N library of novel antimicrobial peptides where 97% showed activity, with top candidates achieving single-digit micromolar potency against critical-priority multidrug-resistant pathogens. Third, to demonstrate that EDEN captures inter-genomic features, we design a gigabase-scale microbiome with over 94,000 synthetic metagenomic assemblies, including prophage genomes and correct cross-species metabolic pathway completions. The EDEN-generated synthetic microbiome covers 9,067 species with a biome-specific taxonomic accuracy of 99%. Over 1,500 of the generated species were outside the fine-tuning dataset while retaining the correct microecological properties and biome association, thus significantly expanding genetic and taxonomic diversity. Together, these results establish a new strategic direction for AI-programmable therapeutics, in which a single foundation model architecture designs candidate therapeutics across diverse modalities and disease areas. This suggests that the combination of billions of years of evolutionary data with specific therapeutic records offers a clear, scaling-driven path to making therapeutic design a predictable engineering discipline. O_FIG O_LINKSMALLFIG WIDTH=141 HEIGHT=200 SRC="FIGDIR/small/699009v2_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@68c20borg.highwire.dtl.DTLVardef@19b8e8corg.highwire.dtl.DTLVardef@1ab9362org.highwire.dtl.DTLVardef@1592cb7_HPS_FORMAT_FIGEXP M_FIG C_FIG

genomics↗

TSTScope Unifies Single-Cell Multi-Omics to Identify Functional T Cell States Predictive of Immunotherapy Response

Immune checkpoint blockade (ICB) can produce durable responses in cancer, but reliable predictors of benefit are still lacking. CD8 tumor-specific T cells are essential for ICB efficacy, yet it remains unclear which functional states of these cells determine therapeutic success. To address this, we developed TSTScope, an interpretable deep learning framework that integrates single-cell transcriptomic and T-cell receptor sequencing data to generate unified representations of CD8 T-cell identity. By applying TSTScope to non-small cell lung cancer (NSCLC) datasets, we characterized the gene programs defining tumor specificity and computationally inferred a population of potential TSTs. Crucially, we demonstrate that clinical response is not a product of TST abundance, but is instead governed by their functional state. We derived the MPR score, a metric capturing this functional potential, which proved to be a robust predictor of treatment outcomes. In an independent validation cohort, the MPR score significantly outperformed established biomarkers. Collectively, TSTScope identifies a distinct functional state of tumor-specific T cells as a primary determinant of ICB efficacy, providing both a mechanistic framework and a potent tool for precision immunotherapy.

bioinformatics↗

Distinct flagellins differentially fine tune biofilm initiation via flagellar stator-associated proteins

Helicobacter pylori relies on flagellar motility to colonize the gastric niche. While H. pylori flagella serve multiple functions, including swimming motility, it has recently been appreciated that flagellar basal body properties also contribute to biofilm initiation. The H. pylori flagellar filament contains two spatially segregated flagellins, FlaA and FlaB, but their roles in the biofilm initiation process remained undefined. Here, we confirmed that both FlaA and FlaB are required for optimal motility, but found that they exert opposite effects on biofilm initiation: cells without FlaA display low biofilm initiation, while cells without FlaB display elevated initiation. To understand the molecular basis for this divergence, cryo-electron microscopy (cryo-EM) analysis of native flagellar filaments revealed that FlaA and FlaB have distinct supercoiled waveforms. We confirmed these intrinsic waveform curvatures by analyzing filaments from {Delta}flaA and {Delta}flaB mutants, providing a physical basis for their functional specialization. Genetic epistasis experiments further demonstrated that the enhanced early biofilm formation of the {Delta}flaB mutant depends on PilO, a stator-associated component of the flagellar motor recently shown to drive a reciprocal biofilm and motility response. Our findings establish that H. pylori has developed functionally specialized flagellins that work with motor-dependent signaling to dynamically balance surface colonization and motility.

microbiology↗

A Comprehensive Treatment-Induced Resistance Atlas of Glioblastoma Reveals a Fibrotic Niche Shielding the Tumor from Immunotherapy

Therapeutic resistance in IDH-wildtype glioblastoma (GBM) is driven by an intricate interplay between cellular plasticity and protective microenvironmental niches. By constructing the GRIT-Atlas--a massive transcriptomic compendium encompassing nearly one million high-quality single cells across 17 cohorts--and cross-analyzing it with 48 independent Visium spatial transcriptomics sections, we discover a recurrent "Spatial Resistance Triad" comprising mesenchymal-like (MES-like) malignant cells, myeloid-derived suppressor cells (MDSCs), and collagen-secreting cancer-associated fibroblasts (CAFs). This large-scale spatial integration reveals that the triad physically fortifies microvascular proliferation (MVP) and pseudopalisading necrosis (PAN) niches. To substantiate these findings at true single-cell resolution, we deploy high-plex spatial molecular imaging (CosMx SMI) to map over 400,000 single cells across a clinical validation cohort. This high-resolution architecture firmly validates the triads geography and, critically, provides structural gradient evidence unmasking a spatiotemporal continuum wherein PAN emerges as a direct functional consequence of MVP advancement and subsequent vascular collapse. Mechanistically, stromal CAF-derived COL6A1 engages CD44 receptors on MES-like cells to accelerate a potent neural stemness cascade within these protective domains. Utilizing a blood-brain barrier (BBB)-penetrant multi-library compound screen, we identify Lacidipine as a multi-modal stromal disruptor that successfully silences CAF activation and halts matrix secretion. Incorporating Lacidipine into the standard chemo-immunotherapy backbone (TMZ + anti-CSF1R) completely dismantles the protective desmoplastic matrix niche, forcing a profound collapse of intracranial tumor burden and dramatically extending overall survival in orthotopic models. Collectively, our study leverages unprecedented single-cell and spatial scale to provide a definitive blueprint of GBM therapeutic evasion, establishing matrix-targeted intervention as a mandatory prerequisite for successful glioblastoma eradication.

cancer biology↗

Systematic microcircuit reconfigurations underlie early experience-induced visual cortical plasticity

Experience-dependent reorganization of neuronal connections during a postnatal critical period (CP) is essential for the functional maturation of developing primary visual cortex (V1). However, the systematic reconfigurations across microcircuit synapses underlying this plasticity were poorly defined. Paired recording from excitatory pyramidal cells and other three major interneurons (INs) within layers 2/3 and 4 microcircuits in the developing mouse V1 monocular region, we find that monocular deprivation (MD) for one-day during the CP transiently reconfigures local excitatory synapses on inhibitory INs that specifically express parvalbumin (PV) or vasoactive intestinal peptide (VIP), whereas prolonged MD primarily strengthens inhibitory transmission from somatostatin (SST)-expressing INs to PV-INs and VIP-INs in both layers. A physiologically based microcircuit model suggests these coordinated synaptic changes in specific inhibitory neurons mediate differential MD-induced plasticity in visual responses across layer 2/3 neurons in vivo. Together, our findings define a systematic microcircuit basis for cortical critical-period plasticity. HighlightsO_LIInhibitory-to-inhibitory connectivity is conserved in cortical layer 2/3 and 4 circuits. C_LIO_LIEarly experience sequentially modifies excitatory and dis-inhibitory synapses in both layers. C_LIO_LISystematic microcircuit reconfigurations underlie plasticity of cortical visual responses. C_LIO_LIModeling suggests a pivotal role of disinhibitory microcircuits in mediating the plasticity. C_LI

neuroscience↗

Acoustoluminescence in Transition Metal and Rare Earth Oxides Beyond 1800 nm for In Vivo Imaging

Acoustoluminescence (AL) is promising for molecular imaging in living tissue, but efficient acousto-optic conversion remains challenging due to the [~]108 times difference in quantum energy between phonons (1 MHz ultrasound) and photons (visible or near-infrared light). Here, we report AL in transition metal oxides (TMOs) and rare earth oxides (REOs) at wavelengths beyond 1800 nm in the short-wave infrared (SWIR) or near-infrared II (NIR-II, 1000-3000 nm) window, under ultrasound excitation at power densities 100-150-fold lower than those required for sonoluminescence in liquids. High-temperature N2/H2-mixed gas reduction was demonstrated as a safe and efficient method to regulate the AL spectra and brightness of TMOs and REOs. TMOs exhibited broadband NIR-II AL emission. Intrinsic emission peaks of rare earth ions and non-conventional luminescence were observed in the AL spectra of REOs under ultrasound excitation. NIR-II AL imaging enabled twice the penetration depth of fluorescence imaging. We developed a scanning focused ultrasound AL imaging system for in vivo tumor imaging through the intact hindlimb, achieving acoustic resolution and penetration depths exceeding one centimeter.

bioengineering↗

Biosynthesis, Structure, and Antibiotic Properties of Gelatinamin A, a Triculamin-like Lasso Peptide

Lasso peptides are structurally unique natural products endowed with high thermal and proteolytic stability, making them attractive as scaffolds for drug discovery. Recently, a new class of lasso peptides containing a second macrocycle, formed between a lysine sidechain and the C-terminus, was discovered, resulting in an even more compact architecture. Here, we report the first NMR structure of the class V lasso peptide, gelatinamin A. Using heterologous expression of the gelatinamin biosynthetic gene cluster (BGC) in Bacillus subtilis, we delineated the biosynthetic pathway through targeted gene deletions. We expressed and characterized the predicted transpeptidase, GelP, that catalyzes the formation of an isopeptide bond between Lys2 and the C-terminus and mediates the reversible conversion of gelatinamin B to gelatinamin A. In addition, we characterize GelT, an N-acetyltransferase that inactivates lasso peptide antimicrobial activity by acetylating a key lysine residue. Furthermore, we demonstrate that gelatinamin is highly potent against several important pathogens and that the activity is strongly bicarbonate-dependent. Finally, we propose a complete biosynthetic pathway for gelatinamin. The structural insight of gelatinamin A and the functional characterization of GelP provide the foundation for future discovery of class V lasso peptides and for engineering transpeptidases to modify other lasso peptide scaffolds.

biochemistry↗

Rapid ecosystem collapse and biofilter formation following seabed methane leakage

Globally, vast amounts of methane are trapped within subseafloor gas hydrates. Recent evidence suggests that climate- and human-induced perturbations can destabilize gas hydrates and trigger methane release, yet the response of deep-sea ecosystems to these changes remains poorly understood. Here we show multi-year in situ monitoring of seabed ecosystem following methane leakage induced by hydrate exploration activities. Within just two years, benthic microbial and eukaryotic diversity in the affected areas declined significantly, while microbial and macrofaunal abundance increased. Integrated geochemical and omics analyses reveal the rapid successional shift of seabed ecosystem to a novel chemosynthetic system. Aerobic methanotrophs (Methyloprofundus) co-established with unexpectedly fast growing anaerobic methanotrophic archaea (ANME-3), accompanied by a rapid recruitment of opportunistic polychaetes that bioirrigated the seabed to >50 cm depths. The active animal-microbe interactions sustained exceptionally high rates of methane oxidation that utilized diverse electron acceptors. We demonstrate that methane hydrate destabilization can trigger rapid collapse of native seabed ecosystem while driving the formation of an effective methane biofilter consuming this potent greenhouse gas much faster than previously estimated. Understanding seabed ecosystem response and feedback is critical for predicting benthic methane cycling under ongoing global change and for developing sustainable strategies for methane hydrate resource management.

ecology↗

Peri-Head Distance Coding in the Mouse Brainstem

Perceiving object distance in peri-personal space is essential for guiding movement and avoiding danger. During active sensation, distance information is often anchored to the body via touch; yet how early somatosensory circuits extract distance information from tactile inputs remains unclear. Here, we investigate how second-order neurons in the mouse whisker brainstem encode peri-head distance. Using in vivo extracellular recordings in awake mice in a naturalistic wall-passing paradigm, we find brainstem neurons employ two distance-coding schemes: a "proximity" code, where firing increases monotonically as objects approach the face; and a "map" code, where neurons exhibit peak tuning at specific distances to collectively tile peri-head space. The map code outperforms proximity code in population decoding of distance. Perturbation experiments reveal multi-whisker integration and internuclear inhibition contribute to the generation of map-like tuning. These findings highlight a previously underappreciated computational role for brainstem circuits, where inhibition acts as a neural comparator to transform proximity-based sensory inputs into a map-like representation of peri-personal space.

neuroscience↗

FuzzAletheia: Mapping dynamic interaction landscapes of intrinsically disordered protein-ligand complexes to accelerate ligand optimization

Intrinsically disordered proteins (IDPs), representing more than 30% of human proteome, present great challenges in rational drug design due to their highly flexible conformations and dynamic interactions. Though their malfunction is linked to major diseases, IDPs are difficult to target. A critical roadblock lies in hit-to-lead optimization due to their dynamic interactions with hit compounds. Here we developed FuzzAletheia, an IDP-ligand dynamic binding pattern analyzer, to accelerate hit optimization. It analyzes the molecular dynamics simulation trajectories of IDP-ligand to cluster interactions into distinct spatiotemporal stable binding features, which are then visualized and quantified to provide insights into the dynamic binding orientation, strength, and stability. This analysis pinpoints modification sites on hit compounds for optimization. We applied FuzzAletheia to the disordered p53 transactivation domain 1 and its hit compound 1050. Guided by FuzzAletheia analysis, we designed and synthesized six derivatives of 1050 and the most potent compound achieved a 10-fold enhancement in activity, demonstrating its power to accelerate IDP hit optimization.

biophysics↗

MAAD: Multidimensional Antiviral Antibody Database

Antibodies have emerged as central components of therapeutic strategies against viral infectious diseases, functioning as key effectors in both prevention and treatment. While traditional antibody discovery has relied heavily on high-throughput screening, the field is now shifting toward rational antibody design, which requires integrative insights into sequence-structure-function relationships. However, the absence of a standardized and well-annotated antibody database integrating these multidimensional features hampers systematic exploration, cross-pathogen comparison, and rational antibody design. Here, we introduce a "Multidimensional Antiviral Antibody Database" (MAAD; http://www.xxx), a curated platform dedicated to antibody nanobody and single-chain variable fragment targeting three high-impact RNA virus families, Coronaviridae (SARS-CoV-1, SARS-CoV-2, MERS-CoV), Orthomyxoviridae (influenza virus), and Pneumoviridae (respiratory syncytial virus, human metapneumovirus), due to the large, high-quality datasets accumulated in recent years. MAAD further incorporates a suite of interactive analysis modules, including CDR annotation, similarity-based CDR3 sequence analysis, V/J gene usage profiling, sequence-based clustering and structure-based antigen-antibody interfaces residues with per-site entropy and mutation rate profiling. These features enable in-depth exploration of antibody sequence characteristics, thereby facilitating functional and structural insights for rational antibody design. Together, by bridging antibody sequence, structure and function, MAAD offers an open and standardized platform that advances comparative antiviral research and supports therapeutic antibody discovery.

bioinformatics↗

Revisiting Humidity Ramp Protocols for Assessing Human Heat Tolerance

BackgroundHumidity ramp protocols are widely used to estimate human heat tolerance limits, yet reported critical environmental limits (CELs) vary markedly across studies. Whether this variability reflects true physiological differences or systematic methodological artifacts related to ramp design remains unresolved. MethodsWe combined first-order thermal modeling with controlled physiological trials to examine how ramp temporal structure (step dwell time) influences apparent rectal temperature (Tcr) inflection points. Twenty-six healthy young adults completed randomized trials at 42 {degrees}C under two protocols: an aggressive-ramp (30-min equilibration followed by humidity increases every 5 min) and a slow-ramp (4-h equilibration followed by hourly humidity increments). CELs derived from ramp protocols were further evaluated using prolonged fixed-condition exposures in independent cohorts (n=14 per sex). ResultsShort-step dwell times ({Delta}t/{tau} <<1) prevented rectal temperature from approaching its step-specific equilibrium, resulting in kinetically constrained, non-equilibrium dynamics and earlier Tcr inflection points. Consequently, aggressive-ramp protocols yielded substantially lower CELs than slow-ramp protocols in both sexes ({approx}3.5 {degrees}C difference). CELs derived from the slow ramp closely matched those obtained from prolonged fixed-condition exposures, whereas aggressive ramps misclassified physiologically compensable conditions as uncompensable. ConclusionRapid humidity increments systematically underestimate heat tolerance because environmental forcing outpaces physiological response kinetics. Accurate CEL determination requires either prolonged fixed-condition exposures (benchmark approach) or sufficiently slow ramps that allow near-equilibrium responses ({Delta}t/{tau} {gtrsim}0.3-0.5).

physiology↗

Drosophila melanogaster model of RVCL-S demonstrates age dependent disease progression

Retinal vasculopathy with cerebral leukoencephalopathy and systemic manifestations (RVCL-S) is a disease that causes deterioration of small vessels, affecting various organs: eyes, brain, liver, and others. The RVCL-S carriers have lower life expectancy. There is no cure available to date. The disease has been linked to mutations in TREX1 gene disrupting its cytoplasmic localization. To facilitate the disease mechanism investigation, we employed model organism D. melanogaster, identified human TREX1 ortholog cg3165, and confirmed its vital significance to flies. Then, we expressed human TREX1 and its mutant form TREX1 V235Gfs in flies and used optical coherence microscopy (OCM) to monitor the dynamics of flies vascular system. We detected the relapse of fly dorsal vessel, movement impairment, and reduced longevity in TREX1 V235Gfs-expressing transgenic animals. Vascular deterioration and shorter life span recapitulate the RVCL-S manifestations in humans. We have established a robust quantitative Drosophila RVCL-S phenotypic system that can potentially serve as a screening platform for drug discovery and drug targets identification.

genetics↗