Search Results (73)

Background: Fibrolamellar carcinoma (FLC) is a rare liver malignancy affecting adolescents. FLCs harbor a DNAJB1–PRKACA gene fusion that combines heat shock protein DNAJB1 with the catalytic subunit of protein kinase A. Surgery with systemic therapy provides 5-year survivals of 30–50%, but advanced disease remains largely incurable. Three-dimensional explants from 41 FLC patients were interrogated for drug sensitivity, resistance, and synergy against cytotoxics, targeted agents, and signal transduction inhibitors. Methods: Sterile specimens from histologically confirmed FLC patients were analyzed by Ex Vivo Analysis of Programmed Cell Death (EVA/PCD™) in a CLIA-licensed laboratory. Following mechanical and enzymatic disaggregation, explants underwent 72 h drug exposure. LC₅₀ values were derived from five-point dose–response curves and compared with a database of over 10,000 human tumor analyses. Synergy was assessed by combination index. In parallel, targeted metabolomic profiling was performed in five FLC patients using tandem MS/MS. Results: Forty-one samples were analyzed. Of 24 drugs selected, tumor-cell yields were adequate for testing in 18 (75%). Single-agent activity favored vorinostat, followed by phenformin and 6-diazo-5-oxo-L-norleucine. Combinations favored gemcitabine plus oxaliplatin (GEMOX) and 5-FU plus interferon. Metabolomic analysis identified distinct signature consistent with mitochondrial dysfunction and altered polyamine metabolism. Conclusions: The present findings are exploratory, and hypothesis-generating and should not be interpreted as evidence of clinical efficacy. Prospective clinical validation and mechanistic studies will be required to further define the therapeutic relevance of these observations in fibrolamellar carcinoma. Full article

A robust analysis of water use in major food production systems is crucial for improving their productivity and sustainability in water-scarce arid and semi-arid regions like Punjab (India) facing the depletion of groundwater resources. This study aimed to assess blue water use and blue water productivity in dairy farming systems across different farm sizes in Punjab. Comprehensive monitoring and assessment of water use over a full year (from July 2022 to June 2023) was conducted on 24 dairy farm sheds in Punjab, revealing significant variability in their blue water use (measured in L per adult animal per day) and blue water productivity quantified as kg of fat- and protein-corrected milk (FPCM) produced per m³ of the blue water consumed. The variability was influenced by factors such as livestock species, farm size (medium with 15–25 livestock, large with 25–100 livestock, and commercial with >100 livestock), bathing and servicing routines, and energy use patterns. The average dairy livestock total blue water consumption varied from 112 ± 14 to 131 ± 19 L per adult animal per day, with 20–40% higher livestock drinking water and about six times higher livestock bathing and serving water used during the summer months. Interestingly, a large share (45%) of the average total blue water consumption is contributed by indirect water consumption via the use of energy (electricity and diesel) in dairy farm sheds. Dairy milk blue water productivity was quantified higher, ranging from 154 ± 11 to 225 ± 59 kg FPCM per m³ in buffalo- and crossbred cattle-based dairy farm sheds. However, indigenous cattle showed a lower blue water productivity ranging from 56 to 97 kg FPCM per m³, reflecting their lower milk yields and limited use of intensified management practices. The state-level water scarcity footprint (WSF) of Punjab dairy farm sheds was quantified at 4870 million m³ world-eq, which showed a significant spatial variation among Punjab districts. However, the results of this study offer novel seasonally and spatially disaggregated benchmarks of blue water consumption, blue water productivity, and the water scarcity footprint of Punjab’s dairy farming sheds. This new information is crucial for the development of locally calibrated and validated models for improving the water productivity and sustainability of dairy farming across Punjab and other similar arid and semi-arid regions in Southeast Asian countries. Full article

Enhancing plant protein structure, functionality, and digestion-associated bioactivity is pivotal to advancing sustainable food applications. In this study, a controlled thermal treatment was applied to Xanthoceras sorbifolium seed meal protein (XSMP) to characterize alterations in structural features, functional performance, and digestion-related bioactivity. Structural analyses showed that moderate heating induced partial unfolding and disaggregation, leading to reduced particle size and improved colloidal stability, with optimal performance observed at 65 °C. Accordingly, foaming capacity and emulsifying activity index reached their highest values under moderate heat pretreatment (71.43% and 27.21 m²/g, respectively). Simulated in vitro gastrointestinal digestion revealed that moderate heat pretreatment enhanced protease accessibility and was associated with increased formation of low-molecular-weight fragments. As a result, digestion products from optimally treated XSMP exhibited significantly enhanced antioxidant activities during the intestinal phase, including higher reducing power, Fe²⁺-chelating capacity (up to 51.21%), and lipid peroxidation inhibition (82.83%). In contrast, insufficient unfolding at lower temperatures or excessive aggregation at higher temperatures reduced the susceptibility to digestive proteases and the associated functional performance. These findings demonstrate that controlled heat treatment provides a simple and eco-friendly strategy to enhance the functional potential of XSMP, supporting its application as a functional protein ingredient. Full article

Polyoxometalates (POMs) exhibit significant potential for application in Alzheimer’s disease (AD) therapeutics owing to their inherent chemical and physical properties and structural tunability. Through transition metal substitution, functional modification, and the construction of POMs-based nanocomposites, POMs can precisely recognize and effectively modulate various key pathogenic proteins involved in Alzheimer’s disease. They can also intervene in disease progression through multiple mechanisms, including inhibition of Aβ aggregation, disaggregation of amyloid-β (Aβ), scavenging of reactive oxygen species (ROS), hydrolytic activity, and modulation of enzyme function. In addition, due to their outstanding physicochemical properties, the application of POMs in phototherapy has emerged as a significant direction in AD treatment research. This review systematically summarizes recent advances from 2011 to 2025 in POMs targeting key pathogenic proteins in AD, comprehensively analyzes their specific mechanisms of action across different therapeutic contexts, highlights their significant advantages and broad potential in AD treatment, and provides new insights for the future structural design, functional optimization, and clinical translation of POMs. Full article

Acinetobacter baumannii relies on biofilms for antibiotic resistance, but the role of planktonic aggregates in drug tolerance is uncharacterized. We studied 103 clinical isolates to explore how the RND efflux pump gene adeG regulates aggregation. Non-MDR strains (with RND deletions) formed aggregates more frequently (13.79%, 4/29) than MDR strains (1.35%, 1/74), driven by residual RND efflux activity (not just deletions). adeG deletion induced 1–2 mm aggregates in a strain with combined adeR/ΔadeABC defects (via upregulated adhesion genes/hydrophobicity) but not in one with only ΔadeC. Aggregates boosted antibiotic tolerance (2–4-fold higher survival vs. disaggregated/parental strains) via metabolic dormancy (5-fold lower ATP), maintained growth in human serum, and promoted persistent bacteremia in immunosuppressed mice. Proteinase K disrupted aggregates, confirming protein matrices’ role. These findings identify planktonic aggregates as pivotal adaptive and virulence-related targets for combating refractory non-MDR A. baumannii infections while also revealing an association between adeG-related genetic contexts and aggregate formation in the bacterium. Full article

The rapidly developing food industry necessitates the efficient use of raw materials, which can be achieved through the production of functional ingredients with high nutritional value. Secondary fish raw materials generated during the filleting of Atlantic cod (Gadus morhua), including vertebral bones with residual muscle tissue, skin, tails, and fins, represent a promising source of both biologically active compounds and highly digestible protein substances. The aim of this study was to evaluate the properties of protein hydrolysates obtained from secondary Atlantic cod raw materials by conventional enzymatic hydrolysis and combined enzymatic-ultrasonic treatment. The best results were achieved at a power of 320 W and a treatment duration of 3.5 min prior to the addition of the enzyme preparation (Protozyme C). The application of ultrasound enhanced the degree of hydrolysis by 4–5% while simultaneously reducing the amount of enzyme used. Electrophoretic analysis demonstrated a predominance of smaller peptides in the 10–15 kDa range compared to the control sample (43–95 kDa). Infrared spectroscopy confirmed structural changes in the samples under study, manifested in an increase in the number of terminal groups and partial disaggregation of the peptide mixture. Particle size distribution analysis revealed a more uniform distribution and a decrease in the median particle size in samples with ultrasonic pretreatment. The safety and antioxidant activity assessment did not show any toxic effects, but manifested a significant increase in antioxidant indicators (2.5–3.2 times) compared to the control sample. The results obtained show the enzymatic-ultrasonic treatment to be promising for the integrated processing of fish raw materials and the production of functional food ingredients with improved properties. Full article

In our previous study, we demonstrated that a standardized ethanol extract of Perilla frutescens var. acuta (PE) alleviates memory deficits in an Alzheimer’s disease mouse model by inhibiting amyloid β (Aβ) aggregation and promoting its disaggregation. However, the extent to which PE exerts additional cognitive benefits independent of Aβ pathology remained unclear. Here, we aimed to evaluate the effects of PE on synaptic plasticity and learning and memory functions. Male ICR mice were used, and cognitive impairment was induced by scopolamine administration. PE was orally administered at doses determined from previous studies, and cognitive performance was assessed using the passive avoidance, Y-maze, and Morris water maze tests. In parallel, hippocampal slices were employed to examine the effects of PE on synaptic plasticity. PE (100 and 300 μg/mL) significantly enhanced long-term potentiation (LTP) in a concentration-dependent manner without altering basal synaptic transmission. This facilitation of LTP was blocked by scopolamine (1 μM), a muscarinic acetylcholine receptor (mAChR) antagonist, and IEM-1460 (50 μM), a calcium-permeable α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (CP-AMPAR) inhibitor, indicating the involvement of mAChR and CP-AMPAR pathways. In vivo, PE (100, 250, and 500 mg/kg) treatment improved memory performance across all behavioral tasks and upregulated hippocampal synaptic proteins including GluN2B, PSD-95, and CaMKII. Collectively, these results demonstrate that PE ameliorates scopolamine (1 mg/kg)-induced cognitive impairment by enhancing synaptic plasticity, likely through modulation of mAChR, CP-AMPAR, and NMDA receptor signaling. These findings highlight the therapeutic potential of PE for memory deficits associated with cholinergic dysfunction. Full article

Heat shock proteins belong to a highly conserved family of chaperone proteins, and in addition to their participation in the regulation of cellular proteostasis (folding of polypeptides and proteins, disaggregation of incorrectly folded peptides, and participation in autophagy processes), also play a significant immunomodulatory role in both innate and adaptive immunity. Changes in the HSP level, both downwards (e.g., in neurodegenerative diseases) and upwards (e.g., autoimmune, oncological diseases), underlie the pathogenesis of many somatic and oncological pathologies. In this review, we consider the main physiological mechanisms of HSP level regulation and also analyze pharmacological, genetically engineered methods of modulating the chaperone level, citing the advantages and disadvantages of a particular method of influence. In conclusion, modulation of the HSP level, according to numerous preclinical studies, can have a significant impact on the course of various pathological conditions, which, in turn, can be used to develop new therapeutic approaches, when the effect on the level of chaperones can be used as monotherapy or as an adjuvant method of action. Full article

Helicobacter pylori (H. pylori), a prevalent human pathogen affecting nearly half the global population, is a major contributor to chronic gastritis, peptic ulcer, and gastric cancer. H. pylori develops biofilms (BFs) allowing bacteria to evade the immune response. Differences in composition between planktonic and biofilm cells influence the host’s immune response, yet the specific biofilm components modulating this response remain uncharacterized. Considering the above, this study evaluated the effect of in vitro-generated H. pylori BF on the antibacterial activity of neutrophils. This work utilized sonication to obtain disaggregated H. pylori BF (d-BF-Hp) to challenge human neutrophils, assessing their bactericidal and phagocytic activity against Staphylococcus aureus. S. aureus survival in the presence of neutrophils was enhanced by 10 μg/mL of d-BF-Hp’s protein. Conversely, S. aureus survival was significantly lower at 30 µg/mL compared to 10 µg/mL d-BF-Hp. Furthermore, 10 and 30 µg/mL of d-BF-Hp significantly reduced the neutrophil phagocytosis rate. Our findings suggest that d-BF-Hp components diminish neutrophil bactericidal activity, although this effect was not observed at higher d-BF-Hp concentrations. Increased d-BF-Hp concentrations proportionally reduced neutrophil phagocytic capacity. Future work should explore the mechanisms underlying the alteration of neutrophil microbicidal properties. Full article

HSP110 is a ubiquitous chaperone contributing to proteostasis. It has a disaggregation activity and can refold denatured proteins. It can regulate fundamental signaling pathways involved in oncogenesis, such as Wnt/β-catenin, NF-κB and STAT3 signaling pathways. In gastric and colorectal cancer, HSP110 has been detected in the nucleus, and nuclear expression has been associated with the resistance of cells to 5-FU chemotherapy. Nuclear translocation of HSP110 is promoted by the exposure of cells to DNA-damaging agents. In a previous work, we demonstrated that nuclear HSP110 participates in the NHEJ DNA repair pathway by facilitating the recruitment of DNA-PKcs to Ku70/80 heterodimers at the site of DNA double-strand breaks. In the present work, analysis of HSP110s’ nuclear interactome revealed an enrichment of components from SWI/SNF chromatin remodeling complexes. We demonstrate that HSP110 is strongly associated with chromatin in temozolomide- and oxaliplatin-treated cells and directly interacts with the core subunit SMARCC2, thereby facilitating the assembly of SWI/SNF complexes. This work expands upon the role of HSP110, which regulates not only proteostasis but also the assembly of critical nuclear macromolecular complexes involved in the adaptive stress response. Full article

The growing demand for plant-based protein alternatives has driven interest in protein modifications to enhance their functional properties in food applications. Enzymatic cross-linking using laccases derived from Rhus vernicifera (LR) and transglutaminase (TG) offers a promising strategy to enhance protein solubility, emulsifying properties, and foaming properties of food proteins. This study varied the enzymatic reaction conditions, including enzyme concentration, pH, temperature, incubation time, and ferulic acid addition, for the most effective cross-linking between proteins in lupin flour (LF) and soy protein isolate (SPI), resulting in changes in physicochemical and functional properties of the cross-linked proteins. LR-induced cross-linking in lupin and soy proteins was most favourable at 142.5 U/100 mg protein, pH 6, and 20 °C, where ferulic acid enhanced cross-linking efficiency with prolonged incubation (20 h). TG-induced cross-linking in lupin and soy proteins was most favourable at 1.25 U/100 mg protein, pH 6 and 30 °C, where high-molecular-weight aggregates were observed. Cross-linking modified protein surface characteristics, increasing ζ-potential and particle size due to protein aggregation, while ferulic acid further enhanced polymerisation. Morphological analysis revealed a porous powder structure across all samples with increased porosity in cross-linked samples as evidenced by the predominance of small fragments within the particles. Prolonged incubation led to partial disaggregation in LR-treated samples unless they were stabilised by ferulic acid. Under mild conditions (1 h, pH 6, 20 °C), LR and ferulic acid-added samples showed minor and significant improvements in protein solubility and foaming stability, respectively. Additionally, a significant increase in foaming ability was observed in ferulic acid-added LR samples after prolonged incubation (20 h), compared to the corresponding control. In contrast, prolonged incubation (20 h) or TG treatment had a lower foaming stability compared to the mild LR treatment. Emulsifying ability and emulsion stability showed limited variation across treatments. These findings suggest that cross-linking conditions influence specific functional properties, highlighting the need for further optimisation to achieve desired protein functionality in food applications. Full article

Hsp70, a 70 kDa molecular chaperone, plays a crucial role in maintaining protein homeostasis. It interacts with the DnaJ family of co-chaperones to modulate the functions of client proteins involved in various cellular processes, including transmembrane transport, extracellular vesicle trafficking, complex formation, and proteasomal degradation. Its presence in multiple cellular organelles enables it to mediate stress responses, apoptosis, and inflammation, highlighting its significance in disease progression. Initially recognized for its essential roles in protein folding, disaggregation, and degradation, later studies have demonstrated its involvement in several human diseases. Notably, Hsp70 is upregulated in multiple cancers, where it promotes tumor proliferation and serves as a tumor immunogen. Additionally, epichaperome networks stabilize protein–protein interactions in large and long-lived assemblies, contributing to both cancer progression and neurodegeneration. However, extracellular Hsp70 (eHsp70) in the tumor microenvironment can activate immune cells, such as natural killer (NK) cells, suggesting its potential in immunotherapeutic interventions, including CAR T-cell therapy. Given its multifaceted roles in cellular physiology and pathology, Hsp70 holds immense potential as both a biomarker and a therapeutic target across multiple human diseases. This review highlights the structural and functional importance of Hsp70, explores its role in disease pathogenesis, and discusses its potential in diagnostic and therapeutic applications. Full article

Heat shock proteins (HSPs) are essential molecular chaperones that protect cells by aiding in protein folding and preventing aggregation under stress conditions. Small heat shock proteins (sHSPs), which include members from HSPB1 to HSPB10, are particularly important for cellular stress responses. These proteins share a conserved α-crystallin domain (ACD) critical for their chaperone function, with flexible N- and C-terminal extensions that facilitate oligomer formation. Phosphorylation, a key post-translational modification (PTM), plays a dynamic role in regulating sHSP structure, oligomeric state, stability, and chaperone function. Unlike other PTMs such as deamidation, oxidation, and glycation—which are often linked to protein destabilization—phosphorylation generally induces structural transitions that enhance sHSP activity. Specifically, phosphorylation promotes the disaggregation of sHSP oligomers into smaller, more active complexes, thereby increasing their efficiency. This disaggregation mechanism is crucial for protecting cells from stress-induced damage, including apoptosis, inflammation, and other forms of cellular dysfunction. This review explores the role of phosphorylation in modulating the function of sHSPs, particularly HSPB1, HSPB4, and HSPB5, and discusses how these modifications influence their protective functions in cellular stress responses. Full article

HSP70 chaperones play pivotal roles in facilitating protein folding, refolding, and disaggregation through their binding and releasing activities. This intricate process is further supported by J-domain proteins (JDPs), also known as DNAJs or HSP40s, which can be categorized into classes A and B. In yeast, these classes are represented by Ydj1 and Sis1, respectively. While both classes stimulate the ATPase activity of Ssa1 (yeast HSP70) through the J-domain, only class B JDPs possess the unique ability to efficiently stimulate Ssa1 in disaggregation processes. The C-terminal EEVD motif of HSP70 plays a crucial role in mediating these interactions by connecting with both client proteins and JDPs. However, the removal of the EEVD motif disrupts the capacity of HSP70 to associate with class B JDPs, and the intricacies of the interaction between these two proteins remain incompletely understood. We employed NMR spectroscopy to investigate the structure and dynamics of the class B J domain protein (JDP) of S. cerevisiae (Sis1) complexed with an EEVD peptide of Ssa1. Our study is based on the extraordinary 70.5% residue assignment of the full-length (352 residues long) Sis1. Our findings revealed that EEVD binds to two distinct sites within the C-terminal domain I (CTDI) of Sis1, to the J domain and to the GF-rich loop located between the J domain and α-helix 6 (a structure identified by this work). We propose that the interaction between EEVD and Sis1 facilitates the dissociation of α-helix 6, promoting a conformational state that is more favorable for interaction with Ssa1. We also employed α-synuclein as a substrate to investigate the competitive nature between EEVD and the client protein. Our experimental findings provide evidence supporting the interaction of EEVD with the client protein at multiple sites and essential insights into the mechanistic cycle of class B JDPs. Full article

Alzheimer’s disease (AD) is a progressive neurodegenerative condition characterized by memory and cognitive decline in older individuals. Beta-amyloid (Aβ), a significant component of senile plaques, is recognized as a primary contributor to AD pathology. Hence, substances that can inhibit Aβ production and/or accumulation are crucial for AD prevention and treatment. Agrimonia pilosa LEDEB. (A. pilosa) (Rosaceae), specifically its aerial parts, was identified in our previous screening study as a promising candidate with inhibitory effects on Aβ production. Therefore, in this study, A. pilosa extract was investigated for its anti-amyloidogenic effects, and its bioactive principles were isolated and identified. The ethanol extract of A. pilosa reduced the levels of sAPPβ and β-secretase by approximately 3% and 40%, respectively, compared to the DMSO-treated control group in APP-CHO cells (a cell line expressing amyloid precursor protein), which were similar to those in the positive control group. In addition, the ethanol extract of A. pilosa also hindered Aβ’s aggregation into fibrils and facilitated the disaggregation of Aβ aggregates, as confirmed by a Thioflavin T (Th T) assay. Subsequently, the active constituents were isolated using a bioassay-guided isolation method involving diverse column chromatography. Eleven compounds were identified—epi-catechin (1), catechin (2), (2S, 3S)-dihydrokaempferol 3-O-β-D-glucopyranoside (3), (-)-epiafzelechin 5-O-β-D-glucopyranoside (4), kaempferol 3-O-β-D-glucopyranoside (5), apigenin 7-O-β-D-glucopyranoside (6), dihydrokaempferol 7-O-β-D-glucopyranoside (7), quercetin 3-O-β-D-glucopyranoside (8), (2S, 3S)-taxifolin 3-O-β-D-glucopyranoside (9), luteolin 7-O-β-D-glucopyranoside (10), and apigenin 7-O-β-D-methylglucuronate (11)—identified through 1D and 2D NMR analysis and comparison with data from the literature. These compounds significantly decreased Aβ production by reducing β- and γ-secretase levels. Moreover, none of the compounds affected the expression levels of sAPPα or α-secretase. Further, compounds 1, 2, 4, 8, and 10 demonstrated a dose-dependent reduction in Aβ aggregation and promoted the disaggregation of pre-formed Aβ aggregates. Notably, compound 8 inhibited the aggregation of Aβ into fibrils by about 43% and facilitated the disassembly of Aβ aggregates by 41% compared to the control group containing only Aβ. These findings underscore the potential of A. pilosa extract and its constituents to mitigate a crucial pathological aspect of AD. Therefore, A. pilosa extract and its active constituents hold promise for development as therapeutics and preventatives of AD. Full article