Search Results (487)

Currently, emerging pharmaceutical contaminants (EPCs) pose a significant risk to aquatic biodiversity on both a global and regional scale. Their accumulation in the environment presents several challenges to both human health and ecological integrity. Among EPCs, cyclophosphamide (COP), a widely used alkylating cytotoxic and immunosuppressive drug in human and veterinary oncology, has become one of the most frequently detected environmental contaminants. It has been reported to impair the immune system, induce oxidative stress, and exhibit genotoxic and cytotoxic effects in various fish species. However, limited research has evaluated its toxicity in the Siamese fighting fish (Betta splendens), an endemic significant aquatic species in Thailand. This study evaluated the acute toxicity of COP in Betta splendens by assessing systemic oxidative stress responses, alterations in gene expression, and localized histopathological changes within both hepatic and renal tissues. The 96 h of median lethal concentration (LC₅₀) value for COP in Betta splendens was determined to be 793.4 mg/L. High-concentration exposure (800 mg/L) induced severe systemic oxidative stress, evidenced by a significant reduction in superoxide dismutase (SOD) activity and a marked elevation in malondialdehyde (MDA) levels across both liver and kidney. At the transcriptomic level, acute COP exposure significantly upregulated pro-inflammatory (interleukin-1β, IL-1β and tumor necrosis factor, TNF-α), cellular stress (heat shock protein 70, HSP70), and apoptotic (caspase-3, Casp3) genes. Semi-quantitative histopathological evaluation revealed severe concentration-dependent structural damage. Hepatic lesions peaked at 800 mg/L characterized by diffuse vacuolation, vascular congestion, and extensive necrosis. Similarly, severe renal damage occurred at 800 mg/L, featuring marked vascular congestion, melanomacrophage aggregation, and widespread tubular necrosis. Overall, these findings demonstrate that acute waterborne COP exposure induces severe hepatotoxicity and nephrotoxicity in Betta splendens driven by oxidative stress, pro-inflammatory signaling, and apoptotic pathways. This study provides essential baseline toxicity thresholds and highlights the physiological risks COP spills pose to tropical freshwater labyrinth fish. Full article

Identifying physiological changes during the transition period is essential for improving the health and productivity of dairy goats. This study evaluated hematological, biochemical, hormonal, oxidative stress, and molecular alterations in Shami goats during the pre-pregnancy, late pregnancy, and early lactation periods. Eighty clinically healthy goats were examined, and blood samples were analyzed for hematological indices, metabolic and hormonal profiles, oxidative stress biomarkers, and relative expression of genes associated with energy metabolism, antioxidant defense, inflammation, and autophagy. Late pregnancy was characterized by significant (p < 0.05) increases in red blood cell count (RBCs), hemoglobin concentration (Hb), neutrophils, albumin, globulin, urea, insulin-like growth factor-1 (IGF-I), and malondialdehyde (MDA), accompanied by decreased glucose, cholesterol, total protein (TP), antioxidant markers, total leukocyte count, packed cell volume, and monocytes. Early lactation was associated with higher non-esterified fatty acid, triiodothyronine (T3), and thyroxine (T4) levels. Genes involved in lipid mobilization and oxidation, ketogenesis, inflammation, cellular stress, and autophagy; sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor alpha (PPARA), carnitine palmitoyltransferase 1a (CPT1A), 3-hydroxy-3-methylglutaryl-coenzyme a synthase 2 (HMGCS2), cluster of differentiation 36 (CD36), lipase E (LIPE), protein kinase amp-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1), haptoglobin (HP), interleukin 6 (IL6), heat shock protein 70 (HSP70), heme oxygenase 1 (HMOX1), beclin 1 (BECN1), and autophagy-related protein 5 (ATG5) were significantly upregulated, whereas antioxidant- and glucose transport-related genes nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 1 (GPX1), catalase (CAT), thioredoxin (TXN), and solute carrier family 2 member 4 (SLC2A4) were downregulated during the transition period. The results indicate well-orchestrated metabolic and molecular adaptations that can be employed as biological indicators to track the physiological status of Shami goats. These findings fulfilled the study objective and identified potential biomarkers of the transition period in Shami goats. Full article

Heat shock protein 70 (HSP70) regulates the stress tolerance and reproductive ability of insects. In this study, the SfHSP70-7 gene was cloned and identified from the rice pest Sogatella furcifera. SfHSP70-7 encodes a typical cytoplasmic HSP70 protein that has a conserved functional domain and is highly similar to the HSP70 homologous protein of Laodelphax striatellus. SfHSP70-7 was found to be widely expressed across all developmental stages, with the highest levels in nymphs and female adults. In terms of tissue distribution, it was particularly abundant in the gut and ovaries. Its expression was strongly induced by high/low temperatures and three insecticides (triflumezopyrim, sulfoxaflor, and imidacloprid), with the maximum induction under heat and triflumezopyrim stress. RNA interference (RNAi) efficiently silenced SfHSP70-7 and significantly increased the susceptibility of S. furcifera to triflumezopyrim and sulfoxaflor, but it had no effect on susceptibility to imidacloprid. RNAi knockdown of SfHSP70-7 markedly reduced survival by 45.5% (30 °C) and 38.9% (35 °C) under heat stress. In addition, gene knockdown can damage the reproductive performance of females by reducing oviposition and egg hatchability. These results indicate that SfHSP70-7 is involved in regulating the heat tolerance, insecticide adaptability, and reproductive regulation mechanism of S. furcifera, providing a potential target for pest control. Full article

Assessment of the stability of inflammatory parameters in bovine blood samples is important because it enables evaluation of the impact of preanalytical factors on biomarker preservation, standardization of sample processing and storage conditions, as well as reliable and consistent interpretation of results in diagnostic and research studies. The aim of the work was to evaluate the stability of the most common inflammatory markers in the blood of cows in early lactation. The influence of the serum–clot contacts duration and the storage of the separated serum in 24 cows, at 25 °C and 4 °C, in intervals of 0–24 h, was examined. Concentrations of interleukin 1 and 6 (IL-1, IL-6), tumor necrosis factor α (TNF-α), interferon γ (IFN-γ), haptoglobin (Hp), serum amyloid A (SAA) and extracellular heat shock protein 70 (eHsp70) were determined by ELISA method, stability was assessed by ANOVA analysis and maximum permissible instability method. Stability was determined over a range of 2–15 h for IL-1, 8–24 h for IL-6, 9–13 h for TNF-α, 10–20 h for IFN-γ, 20–24 h for Hp, 15–24 h for SAA, and 9–12 h for eHsp70. The stability was strongly dependent on temperature and sample type, with storage at 4 °C providing the highest stability in most cases, while differences between serum and serum–clot samples were analyte-specific and less consistent compared to the effect of temperature. Full article

Heat shock proteins of the 70 kDa family (HSP70s) are essential molecular chaperones that preserve proteostasis by assisting protein folding, transport, and degradation. Although the core HSP70 architecture is deeply conserved, the degree and functional significance of sequence divergence across vertebrates remain incompletely understood. Here, an integrative comparative analysis of HSP70 proteins from ten representative vertebrate species spanning mammals, birds, amphibians, and teleost fish was performed. Multiple sequence alignment, phylogenetic reconstruction, motif discovery, entropy-based conservation profiling, hydrophobicity analysis, and structural mapping reveal a strikingly conserved ATPase domain alongside a more variable substrate-binding domain and C-terminal region. Multiple Expectation Maximization for Motif Elicitation (MEME) motif analysis identifies both universally conserved motifs and lineage-specific elements, highlighting functional constraint as well as adaptive diversification. Structural projection onto the human HSP70 crystal structure (PDB 5AQV) demonstrates that conserved hydrophobic residues cluster in the protein core, whereas variable residues are predominantly surface-exposed. Together, these findings illuminate how evolutionary pressures shape both the conserved chaperone machinery and flexible regulatory regions of HSP70, and they establish a scalable analytical framework for comparative protein evolution studies. Full article

Type 2 diabetes (T2D) is a chronic metabolic disorder leading to increased cardiovascular risk and vascular dysfunction. Hyperglycemia, a hallmark of T2D, drives hypercontractility, thereby compromising vascular function. Heat shock protein 70 (HSP70) has emerged as an important player in vascular reactivity under physiological conditions via its interaction with calcium mobilization, and in T2D, blocking this protein prevents hypercontractility. Circulating extracellular HSP70 (eHSP70) has also been proposed as a biomarker in chronic diseases, as it can function as a damage-associated molecular pattern (DAMP) to activate the innate immune system and promote low-grade inflammation. Berberine (BBR), a natural alkaloid with anti-inflammatory properties, has been shown to attenuate vascular contraction by modulating intracellular calcium handling. Yet the link between HSP70 and BBR in modulating vascular contraction in T2D remains unknown. Therefore, we investigated whether acute and/or chronic BBR treatment modulates HSP70 to prevent vascular hypercontractility in the T2D mouse model. For acute ex vivo treatment, db/+ and db/db aortic rings were incubated for 30 min with or without the HSP70 inhibitor VER155008, in the presence or absence of BBR or vehicle. For chronic in vivo treatment, db/+ and db/db mice received intraperitoneal BBR injections (10 mg/kg, 3 times per week) and BBR in their drinking water (0.5 mg/mL) for 28 days. Following chronic (4 weeks, in vivo) or acute ex vivo (30 min) BBR treatment, vascular function was assessed in aortic rings isolated from male T2D (db/db) and age-matched non-diabetic (db/+) mice using wire myography. Rings were incubated with or without the HSP70 inhibitor VER155008, in the presence or absence of BBR or vehicle. Overt hyperglycemia and hypercontractility were observed in diabetic animals compared with non-diabetic controls. While acute BBR treatment attenuated vasoconstriction in both diabetic and nondiabetic groups, the combination of BBR and VER155008 produced a stronger inhibitory effect only in the diabetic group. Chronic BBR treatment prevented aortic hypercontractility in diabetic mice; however, the synergistic effect with VER155008 was no longer observed. Additionally, BBR reduced systemic HSP70 levels. Collectively, these findings indicate that BBR improves vascular smooth muscle cells’ function in T2D, at least in part, through HSP70-dependent mechanisms during chronic treatment. Full article

Objectives: Neuroinflammation is recognized as a significant characteristic of Alzheimer’s disease (AD). Currently, there is a notable absence of effective pharmacological agents to prevent or treat neuroinflammatory processes associated with AD. Heat shock protein 70 (HSP70) is pivotal in the progression of neuroinflammation. In this study, we explored the potential of maltol, a Maillard reaction product derived from red ginseng, as a therapeutic agent for neuroinflammation. Methods: In vitro, HMC3 microglial cell models were developed to examine the regulatory effects of gradient concentrations of maltol (12.5, 25, 50 μM) on the TLR4/MyD88/NF-κB p65 signaling pathway, neuroinflammation, and pyroptosis. Analyses of the GEO database and Gene Set Enrichment Analysis (GSEA) were performed to identify the core targets of maltol, followed by HSP70 gene silencing experiments to validate the targeted regulatory mechanism. Results: Maltol significantly mitigated LPS-induced neuronal damage and cognitive deficits in mice. It effectively suppressed microglia-mediated neuroinflammation and pyroptosis, reversed oxidative stress-induced neuronal ferroptosis, and inhibited neuronal apoptosis. In vitro experiments demonstrated that maltol obstructed TLR4/MyD88 binding, thereby inhibiting NF-κB p65-mediated neuroinflammation and pyroptosis, while also alleviating excessive ROS accumulation to enhance oxidative stress and ferroptosis. Bioinformatics analysis identified HSP70 as a crucial target for the anti-inflammatory and antioxidant effects of maltol. Subsequent gene silencing experiments confirmed that maltol exerted its inhibitory effects on LPS-induced neuroinflammation and pyroptosis in an HSP70-dependent manner. Conclusions: Maltol exhibits significant protective effects against Alzheimer’s disease-related neuroinflammation, oxidative stress, pyroptosis, and ferroptosis through the targeting of HSP70. This study elucidates the molecular mechanisms by which maltol improves neuroinflammatory injury and provides a novel theoretical foundation and therapeutic strategy for the intervention of Alzheimer’s disease neuroinflammation using traditional Chinese medicine. Full article

Background/Objectives: The genes encoding HSPA1A, HSPA1B, and HSPA1L, located in the MHC class III region at 6p21.3–22.1, a region implicated in susceptibility to schizophrenia, are critical regulators of neurodevelopmental processes and contribute to synaptic neuroprotection. This study investigated whether the HSPA1A, HSPA1B, and HSPA1L genes are associated with schizophrenia. Methods: We sequenced the coding regions of HSPA1A, HSPA1B, and HSPA1L from 100 patients with schizophrenia to identify genetic variants. Further, we conducted a genetic association analysis of three SNPs (rs9469057, rs142416335, and rs2075800) in the HSPA1L gene in 519 patients with schizophrenia and 1492 healthy controls from the Taiwan Biobank. We analyzed the function of the HSPA1L protein via immunoblotting. Results: We identified 17 coding variants, including 8 missense and 9 synonymous mutations, in 100 patients with schizophrenia. Three variants (HSPA1L^p.Ala8Pro, HSPA1L^p.Ala8Thr, and HSPA1L^p.Glu602Lys) in the HSPA1L gene did not exhibit any significant differences in allele or genotype frequencies between patients and control subjects. Notably, one ultra-rare missense mutation, HSPA1L^p.Val262Met, was not documented in the control sample in Taiwan BioBank. Immunoblotting revealed HSPA1L^p.Val262Met mutant with decreased protein expression in SH-SY5Y cells compared with the wild type. Conclusions: While common variants in the HSPA1A, HSPA1B, and HSPA1L genes do not seem to be significant genetic risk factors for schizophrenia in this cohort, the ultra-rare mutation, HSPA1L^p.Val262Met, significantly reduces protein expression. These preliminary findings suggest that a potential loss-of-function or reduced expression of the HSPA1L gene may be a predisposing factor contributing to schizophrenia vulnerability in certain individuals. However, the finding should be replicated in other independent samples. The in vitro and in vivo impacts of the associated mutation at the HSPA1L gene on the pathophysiology of schizophrenia are worthy of future investigation. Full article

Background/Objectives: Tordylium trachycarpum Boiss. (Apiaceae) is traditionally used in Kurdish ethnomedicine for the management of gastrointestinal disorders; however, its pharmacological efficacy and safety profile remain insufficiently investigated. This study evaluated, for the first time, the gastroprotective activity and associated antioxidant, inflammatory, and apoptotic responses of the methanolic extract of T. trachycarpum using an ethanol-induced gastric ulcer model in Sprague–Dawley rats. Methods: Preliminary phytochemical screening revealed the presence of phenolics, flavonoids, terpenoids, tannins, coumarins, and glycosides. Acute oral toxicity testing demonstrated no signs of toxicity at doses up to 5 g/kg. Gastric ulceration was induced by absolute ethanol, and animals were pretreated with the extract (250 and 500 mg/kg) or omeprazole (20 mg/kg). Results: The extract significantly decreased the gastric lesion area from 258.50 ± 6.38 mm² in the ulcer control group to 143.70 ± 0.76 mm² and 115.50 ± 0.76 mm², corresponding to ulcer inhibition rates of 44.41% and 55.31%. Additionally, the extract increased mucus production, maintained mucosal structure, and raised stomach pH. Biochemical analysis showed a significant increase in antioxidant enzymes [superoxide dismutase (SOD) and catalase (CAT)] and a reduction in malondialdehyde (MDA) levels, indicating attenuation of oxidative stress. In addition, the extract modulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-10). Blood-based ELISA analysis demonstrated increased expression of heat shock protein 70 (HSP70) and reduced Bax levels, suggesting anti-apoptotic activity. Conclusions: These findings indicate that T. trachycarpum exerts significant gastroprotective activity through antioxidant, anti-inflammatory, and anti-apoptotic mechanisms, supporting its traditional use and highlighting its potential as a natural therapeutic candidate for the management of gastric ulcers. Full article

Coastal ecosystems are increasingly threatened by human activities, highlighting the need for sensitive tools to assess environmental risk. An active biomonitoring approach, using the Mediterranean mussel (Mytilus galloprovincialis), was employed to evaluate anthropogenic chemical contamination in the North Ionian Sea, a still poorly studied area, by comparing mussel health status before (PrePT) and after (PostPT) the peak tourist season. Bioaccumulation of metal(loid)s was quantified in whole organisms. Oxidative stress was assessed in the gills and digestive gland through catalase (CAT), superoxide dismutase (SOD), lipid peroxidation (LPO), and oxidized carbonyl proteins (OMP). Neurotoxicity was evaluated via acetylcholinesterase (AChE) activity, while gene expression of stress-related biomarkers was analysed for metallothioneins (mt10, mt20), sod, cat, Glutathione S-transferase (gst), and Heat Shock Protein 70 (hsp70). Results suggest a progressive contaminant accumulation likely associated with intensified summer anthropogenic activity. Biomarker responses revealed clear activation of oxidative stress, with tissue-specific patterns. The findings confirm the effectiveness of active biomonitoring and multibiomarker approach in assessing coastal water quality and provide valuable baseline data for the management of marine ecosystems. Full article

Cold stress poses a significant challenge to aquatic organisms, affecting their survival, growth, and metabolic processes. This review explores the molecular mechanisms by which fish, crustaceans, and mollusks respond to cold stress, highlighting the shared and species-specific pathways that facilitate adaptation. Common responses to cold stress include modulation of energy metabolism, regulation of oxidative stress, immune responses, and maintenance of proteostasis. In particular, the activation of the adenosine 5′-monophosphate-activated protein kinase (AMPK) and mechanistic target of rapamycin (mTOR) pathways plays a critical role in regulating energy balance and autophagy in response to low temperatures. Furthermore, we examine the specific adaptive mechanisms employed by different groups of aquatic organisms. Fish utilize pathways such as peroxisome proliferator-activated receptor alpha/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPAR/PGC-1α) and fatty acid oxidation to optimize energy utilization and improve cold tolerance. Crustaceans rely on crustacean hyperglycemic hormone (CHH) signaling and AMPK pathway activation, while mollusks employ metabolic suppression and glycogen storage to survive cold exposure. Moreover, the regulation of autophagy and apoptosis, mediated by p53 and cyclin-dependent kinase 1 (Cdk1), ensures the survival of healthy cells under prolonged cold stress, with autophagy maintaining energy homeostasis and apoptosis eliminating damaged cells. This review also discusses the role of molecular chaperones like heat shock protein 70 (HSP70) and the ubiquitin-proteasome system (UPS) in protein homeostasis, highlighting their importance to protect cells under cold stress. The combined action of these molecular pathways allows aquatic organisms to cope with and adapt to cold environments, ensuring cellular integrity and enhancing survival. Future research should focus on integrating molecular, physiological, and ecological approaches to better understand cold tolerance mechanisms and improve aquaculture practices under climate change scenarios. Full article

Heat shock proteins (HSPs) fulfil protective tasks in the whole organism; in pregnant dogs, they are expressed in the ovary, placenta and preimplantation embryo. Our objective was to compare the expression of HSP60 and -70, along with indicators of proliferation and apoptosis, in the non-pregnant and pregnant uterus/placenta and ovaries. Tissues were obtained after ovariohysterectomy and examined by means of immunohistochemistry. There were differences between pregnant and non-pregnant tissues: the expression level of HSP70 during preimplantation in superficial cells was significantly lower than that in early diestrus, with similar results observed for Ki67. The immunosignal for HSP70 was significantly decreased during the postimplantation stage in almost all cell types, whilst the number of HSP60-positive cells did not change. In pregnant animals, the number of Ki67-positive cells significantly increased until the postimplantation stage. In the placenta and trophoblast, the expression of HSP60 and -70 was strong, while no HSP70 signal was detected in endometrial epithelial cells. The caspase 3 immunosignal in the uterus and placenta was generally weak. In the corpora lutea, HSP60, HSP70 and caspase 3 were mainly detected in theca lutein cells, while no signal for KI67 was seen. In follicles, caspase 3 and KI67 expression was low, except in granulosa cells of tertiary follicles and oocytes. We conclude that the different expression of HSPs in pregnant and non-pregnant animals may point towards different regulatory and/or protective tasks. Full article

Coronavirus disease 2019 continues to pose global health challenges, with the pandemic significantly burdening several economies, healthcare systems, and the social lives of individuals. Furthermore, new cases continue to be reported, underscoring the need for therapeutic strategies targeting conserved regions and host–virus interactions. Building on earlier virtual screening for small molecules, all-atom molecular dynamics simulations and binding-free-energy calculations were performed to elucidate how the two previously identified small molecules (NSC36398 and NSC281245) may affect the dynamic behaviour of the interaction between heat shock 70 kDa protein 8 (HSPA8) and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein. Post-MD analyses refined prior docking predictions, where NSC281245 was found to bind tightly to the complex with limited perturbations at the HSPA8-spike protein interaction surface, whereas NSC36398 appeared to induce allosteric-like domain-level destabilisation effects while maintaining stable polar contacts with the protein. Our findings demonstrate the potential of NSC36398 as a promising modulator for disrupting the HSPA8-spike protein complex, which may serve as a structural lead for designing next-generation inhibitors of host–virus interactions. Full article

Sheng Mai San (SMS), a traditional Chinese medicine formula for treating qi and yin deficiency, is widely used in the management of conditions such as cardiovascular diseases and heatstroke. However, its role in mitigating heat stress (HS)-induced liver injury remains underexplored. In this study, a rat model of HS was established under high-temperature and high-humidity conditions, and SMS was administered as an intervention. The pharmacodynamic effects of SMS were comprehensively evaluated through histopathological examination, detection of heat shock protein 70 (HSP70) and heat shock protein 90(HSP90) expression, and analysis of liver function biomarkers (AST, ALT). Meanwhile, oxidative stress indicators were measured using biochemical assay kits (GSH, SOD, CAT, MDA, T-AOC), and transmission electron microscopy was employed to observe mitochondrial ultrastructure, thereby assessing the protective effects of SMS on hepatic oxidative stress and mitochondrial damage induced by HS. In vitro, BRL-3A cells were cultured, subjected to HS, and treated with SMS. Cell viability was assessed using the CCK-8 assay, and changes in mitochondrial reactive oxygen species (ROS) levels, mitochondrial permeability transition pore (MPTP) opening, and mitochondrial membrane potential (MMP) were evaluated using fluorescent probes. The results showed that SMS effectively restored HS-induced histopathological damage in rat liver tissues, reduced serum AST and ALT levels, and downregulated the mRNA expression of HSP70 and HSP90 in liver tissues. Meanwhile, SMS strengthened the hepatic antioxidant system by increasing the levels of GSH, SOD, T-AOC, and CAT, while decreasing MDA content. In vitro experiments confirmed that SMS increased the viability of BRL-3A cells, reduced ROS production, improved MPTP opening/closing regulation, and stabilized MMP. This study provides a clinical reference for its application in treating HS-related conditions in humans and animals. Full article

Background: Liquid biopsy-based biomarkers provide valuable insights into tumor biology, dynamics, burden, relapse prediction and therapeutic responsiveness. The stress-inducible heat shock protein 70 (Hsp70), which is frequently overexpressed in highly aggressive solid tumors and is presented on the cell membrane of tumors but not normal cells, is found in the circulation either as a free protein originating from dying cells or in the context of extracellular vesicles (EVs) that are actively released by viable tumor cells. This study demonstrates the potential value of circulating Hsp70 (eHsp70) levels across multiple solid tumor entities as an entity- and stage-dependent diagnostic biomarker reflecting tumor burden and disease stage. Methods: Circulating eHsp70 levels, as determined using the Hsp70-exo ELISA which detects free and EV-associated Hsp70, in plasma samples collected from patients with different tumor entities (n = 389) prior to the initiation of any oncological therapy and healthy controls (n = 108) between 2021 and 2025, were analyzed retrospectively. Tumor stages were categorized as early, locally advanced, or metastatic. The Kruskal–Wallis test was used for group comparisons and the Receiver Operating Characteristic (ROC) curve was used to evaluate the diagnostic performance of eHsp70 levels. DeLong’s test was used to calculate differences between AUC values. Results: In tumor patients (n = 389), circulating eHsp70 levels were significantly higher than those in healthy controls (n = 108) (Kruskal–Wallis, p < 0.001). eHsp70 levels progressively increased from early-stage to locally advanced and metastatic disease in a stage-dependent manner. Although ROC analysis demonstrated the limited discriminatory performance of eHsp70 levels in early-stage disease (AUC 0.569), increased discrimination was apparent in locally advanced disease (AUC 0.751), metastatic tumors (AUC 0.784) and combined advanced tumor diseases (AUC 0.765; significant by DeLong’s Test comparing early-stage to locally advanced and metastatic tumors), irrespective of the tumor entity with the highest AUC values in metastatic breast cancer (AUC 0.872), sarcoma (AUC 0.861) and non-small cell lung cancer (NSCLC) (AUC 0.835). Apart from minor entity-specific differences, the correlation of eHsp70 levels with the tumor stage remained consistent across all measured tumor entities. Conclusions: Circulating eHsp70 levels are markedly elevated in patients with highly malignant solid tumors and show a consistent, stage-dependent increase across multiple tumor types. These findings suggest that circulating eHsp70, as an indicator of tumor-associated cellular stress and overall tumor burden, represents a valuable biomarker for assessing disease stage, monitoring disease progression, and evaluating therapeutic responses. Full article