BACKGROUND/AIMS:Bone metastases are a frequent and highly debilitating complication of advanced malignancies and are associated with severe pain, structural instability and loss of functional capacity. Radiotherapy, systemic antineoplastic therapy, bone-targeted agents and interventional techniques remain the clinical standard, yet a relevant proportion of patients experience only incomplete or transient pain relief. Non-invasive focal approaches are therefore being explored. Tumor-destructive mechanical impulses (TMI) - an umbrella term for shock-type, high-strain acoustic pulses generated by electrohydraulic, electromagnetic or piezoelectric systems - have a long safety record in musculoskeletal medicine and are increasingly investigated in oncology. We report the palliative use of TMI in patients with painful bone metastases. METHODS:Patient-specific DICOM data were segmented using Simpleware ScanIP® and imported into ANSYS SpaceClaim for geometry preparation, enabling coupled device-tissue simulations under clinical pressure wave forms; the resulting anatomical models were simulated in ANSYS Explicit Dynamics. In parallel, DICOM data were processed using MATLAB®/TABLIN and converted into OnScale® for high-frequency pressure wave propagation analysis, predicting pressure fields, focal volumes and attenuation through bone. The outputs of the numerical solution of the finite element (FEM) propagation models comprise total energy, energy flux density, pulse count and frequency, and the optimal placement of the TMI applicator. TMI was applied within individual healing attempts in patients in whom conventional therapy had failed. RESULTS:In two patients with metastatic prostate carcinoma described in detail, and in thirteen further patients with bone metastases, TMI applied to metastatic bone lesions was followed by relief of periosteal pain. In both index patients, follow-up Ga-68-PSMA PET-CT showed marked regression of the osseous lesions. Treatments were tolerated without device-related complications; in one patient the piezoelectric technique was not tolerated and was replaced by the electrohydraulic technique. CONCLUSION:TMI treatment combines direct mechanotransduction with controlled cavitation, potentially disrupting tumor cells and microvasculature and inducing features of immunogenic cell death. In the patients reported here, palliative TMI treatment was associated with pain relief and radiographic tumor regression, including observations compatible with a possible immunological abscopal response. These observations derive from uncontrolled individual healing attempts under concomitant systemic therapy and require prospective confirmation.
Background/Aims: The aim of this research was to use Mebendazole as an anticancer candidate to reduce the dose of Flutamide and reduce its side effects. Method: In this in vitro study, we evaluated the effect of Mebendazole, Flutamide and Mebendazole-Flutamide combination therapy in LNCaP, DU145 and PC3 cell lines as representatives of human prostate cancer. The assessment includes scratch-wound assay, colony formation assay, flow cytometric analysis of apoptosis and DNA cell cycle, real-time PCR (BAX/BCL2, E-cadherin, N-cadherin, Snail, HIF1α, VEGFC, KLK3, TUBB1 and TUBB3 genes). Results: To determine IC50 levels, cell lines were exposed to different concentration of the drugs. Our data indicated that IC50 values for Mebendazole (55μM) and for 3 cell lines and flutamide (12μM and 10μM) for PC3 and LNCaP/DU145 respectively, with MTT were approved by flow cytometry in a dose and time-dependent manner which was as a consequence of cell cycle arrest at G1/S phase. Furthermore, for the first time, we offered that combination of mebendazole and flutamide produced a greater inhibitory effect on cell viability, colony formation, and migration than either agent alone at the tested concentrations. The combination treatment also increased apoptotic cell populations and upregulated the BAX/BCL2 mRNA ratio and E-cadherin expression in all three cell lines (P<0.01) and downregulated the expression of TUBB1 in DU145 and TUBB3 genes in DU145 and PC3 cell lines (P<0.01). Conclusion: Mebendazole in combination with flutamide reduced dose of flutamide and increased the sensitivity of prostate cancer cells to treatment. Therefore, this combination may represent a promising in vitro strategy that warrants further mechanistic and in vivo investigation.
Background/Aims: Acinetobacter baumannii is an opportunistic gram-negative pathogen and an increasingly important cause of hospital-acquired infections, particularly in intensive care units. Its remarkable ability to rapidly acquire resistance mechanisms, especially against carbapenems, represents a major public health concern. This study aimed to investigate the molecular detection and characterization of OXA-type carbapenemase genes in A. baumannii isolates collected from various clinical sources in Baghdad, Iraq. Methods: Between March and July 2025, 36 non-repetitive A. baumannii isolates were obtained from patients with different infections. Identification was performed using standard biochemical tests, CHROMagar Acinetobacter, and the VITEK 2 system and was confirmed by PCR amplification of the intrinsic blaOXA-51 gene. Antimicrobial susceptibility testing was conducted according to CLSI guidelines. The prevalence of blaOXA-23, blaOXA-24, blaOXA-51, and blaOXA-58 genes was determined by PCR. Selected PCR products were sequenced and subjected to phylogenetic analysis. Results: Extensive antimicrobial resistance was observed among the isolates, particularly to carbapenems, with resistance rates of 83.3% for imipenem and 72.2% for meropenem. High resistance rates were also detected for fluoroquinolones and aminoglycosides, whereas colistin and tigecycline retained comparatively greater activity. PCR screening revealed prevalence rates of 100% for blaOXA-51, 86.1% for blaOXA-23, 69.4% for blaOXA-24, and 47.2% for blaOXA-58. Multiple blaOXA genes were detected in more than half of the isolates, suggesting horizontal gene transfer and local clonal expansion. Phylogenetic analysis demonstrated high similarity between local isolates and international reference strains, supporting the widespread dissemination of resistance determinants. Several nucleotide substitutions were identified within the blaOXA-23 and blaOXA-24 genes. Conclusion: The findings indicate that blaOXA-23 is the predominant contributor to carbapenem resistance among A. baumannii isolates in Baghdad, while blaOXA-24 and blaOXA-58 are also increasingly prevalent. The observed resistance patterns and phylogenetic relationships underscore the importance of continuous molecular surveillance, antimicrobial stewardship, and effective infection control measures to limit the spread of multidrug-resistant A. baumannii. These data contribute valuable regional information to the global understanding of antimicrobial resistance epidemiology.
BACKGROUND/AIMS:Underutilized native legumes may contribute to the development of sustainable, nutrient-dense foods with potential relevance to malnutrition and diet-related non-communicable diseases. METHODS:This study evaluated the nutritional and functional profile of five native Ecuadorian legumes (Cajanus cajan, Lablab purpureus, Phaseolus lunatus baby lima, Phaseolus lunatus big lima, and Vigna unguiculata) through proximate composition, dietary fiber, mineral content, antioxidant capacity, amino acid profiles, amino acid scores (AAS), and phytate molar ratios. RESULTS:Lablab purpureus showed the highest amino acid score (166.30%), followed by P. lunatus baby lima (159.46%) and V. unguiculata (156.82%), and all species exceeded the FAO/WHO indispensable amino acid reference pattern for older children, adolescents, and adults. Cajanus cajan was characterized by high dietary fiber (31.44 g/100 g) and calcium contents (5750 mg/kg), whereas Lablab purpureus and Vigna unguiculata showed higher iron and magnesium contents, together with antioxidant responses associated with phenolic compounds. Although phytate molar ratios suggested potential constraints on non-heme iron bioavailability, the comparatively low phytate ratios indicated lower predicted interference with magnesium availability. Principal component analysis explained 77.6% of the total variance and revealed differentiated nutritional profiles among species. CONCLUSION:These findings support a cautious translational interpretation of underutilized Ecuadorian legumes as differentiated plant food matrices in which phenolic compounds, dietary fiber, amino acids, and minerals may contribute to redox balance, gut barrier physiology, protein adequacy, and micronutrient nutrition.
Background/Aims: Cocoa bean fermentation is a critical step in chocolate production, requiring reliable quality control to verify proper fermentation. Because lysine is formed during fermentation and reflects the underlying biochemical changes, it may serve as a useful indicator of fermentation quality. This study aimed to develop and optimize an enzymatic sensor for the rapid determination of lysine during cocoa bean fermentation. Methods: An enzymatic sensor based on lysine-alpha-oxidase immobilized on a nylon membrane was developed and optimized. Sensor performance was evaluated by calibration, linearity, response time, and application to cocoa protein extracts obtained at different stages of spontaneous fermentation. Lysine concentrations were determined from oxygen consumption measurements. Results: The sensor exhibited a positive linear relationship between oxygen consumption and lysine concentration over the range of 0.3-1.5 mM (R2 = 0.9995). The optimal response time was 5 s, and lysine concentrations were calculated from the slopes of the reaction rates. The immobilized enzymatic sensor enabled rapid and precise determination of lysine in cocoa protein extracts, demonstrating its suitability for monitoring fermentation progress. Conclusion: The developed lysine sensor provides a fast, precise, and environmentally friendly approach for monitoring cocoa bean fermentation with minimal reagent consumption. Its application represents a promising alternative for quality control in the chocolate industry and supports efficient assessment of fermentation through lysine quantification. (c) 2026 The Author(s). Published by Cell Physiol Biochem Press GmbH&Co. KG
Cardiovascular diseases remain the primary driver of global mortality, with advanced age serving the most significant risk factor for their development and progression. Emerging evidence suggests that chronic infections can act as potent catalysts for cardiac decline by prematurely inducing aging phenotypes. Pathogens, including viruses, bacteria, and parasites, that evade host clearance establish a state of permanent inflammaging: a chronic, low-grade inflammatory milieu characterized by persistent cytokine signaling and leukocyte infiltration. This environment directly mirrors the sterile inflammation that drives natural senescence. Mechanistically, chronic infection subverts the heart's homeostatic pathways, triggering cardiomyocyte senescence through the dysregulation of mTOR signaling and the impairment of autophagy. These infections further drive mitochondrial dysfunction and the overproduction of reactive oxygen species (ROS), leading to oxidative DNA damage and metabolic exhaustion within the myocardium. On a structural level, immune subversion, via macrophage polarization and the induction of autoimmunity, accelerates left ventricular hypertrophy, myocardial remodeling, and interstitial fibrosis. By characterizing chronic infection as a modifiable driver of biological aging, we can prioritize anti-infective strategies as essential components of cardiovascular longevity and geriatric care.
Causal therapy has achieved success in the treatment of epithelial tumors, which account for more than 80% of all cancers. Although frequently claimed as breakthroughs, cancer therapy has achieved limited increases in survival of only weeks to several months, and cancer incidence continues to increase while metastasis rates, which are primarily responsible for cancer mortality, remain constant. This reflects an incomplete understanding of carcinogenesis and metastatic progression. Over a 25-year timeframe, the series "Epistemology of the Origin of Cancer" has examined the biological basis of carcinogenesis and metastasis. Part I addressed carcinogenesis, Part II identified the first cancer cell, and Part III described the development of local pre-metastatic niches and traveling cancer satellites. In this fourth part, the conditions required for distant metastasis are discussed, including the sequential development of metastatic niches (MN-1, MN-2, and MN-3), transendothelial migration, dormancy, immune modulation, extracellular matrix remodeling, and metastatic niche maturation. The review proposes that metastatic progression depends on the formation of metastatic cancer satellites consisting of metastatic cancer cells, metastasis-associated fibroblasts (MAFs), stromal components, chemokine coatings, platelets, and neutrophil extracellular traps (NETs), each contributing to immune evasion and dissemination. This sequential distant metastatic niche model provides a biological framework explaining clinical observations including metastatic dormancy, relapse after surgery or anticancer therapy, tumor heterogeneity, and the limited long-term success of current therapeutic approaches.
BACKGROUND/AIMS:The management of multiple myeloma (MM), a formidable hematological malignancy, continues to pose substantial challenges. Schisandrin B (Sch B), a bioactive compound derived from Traditional Chinese Medicine, has demonstrated potent antitumor properties, but its in vivo effects on MM remain unclear. METHODS:Tumor growth was evaluated by measuring body weight, tumor volume, and cell proliferation. Cell cycle progression and apoptosis were analyzed by flow cytometry. ELISA was performed to quantify interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF). Reactive oxygen species (ROS) were detected by immunofluorescence, and Western blotting was used to determine the expression of IL-6, JAK2, phosphorylated JAK2 (p-JAK2), STAT3, and phosphorylated STAT3 (p-STAT3). RESULTS:Sch B markedly inhibited tumor proliferation, induced S-phase cell cycle arrest, and promoted apoptosis. Sch B reduced the expression of IL-6, VEGF, p-JAK2, and p-STAT3 while increasing ROS levels in mice with MM. Moreover, Sch B exhibited a synergistic antitumor effect when combined with bortezomib. CONCLUSION:Schisandrin B represents a promising therapeutic candidate for multiple myeloma, and its antitumor efficacy is further enhanced in combination with bortezomib.
BACKGROUND/AIMS:To present a clinical case of metabolic syndrome (MetS) and to provide a concise pathophysiological overview with emphasis on underlying metabolic and inflammatory mechanisms. METHODS:This study is a case report combined with a focused literature review. We describe a patient with MetS who was evaluated and treated at the Hospital of Vietnam National University in Hanoi, Vietnam. RESULTS:A 20-year-old male presented with headaches and palpitations. Clinical examination revealed obesity (BMI 37.1) with a waist circumference of 111 cm, dyslipidemia (triglycerides/HDL-cholesterol/LDL-cholesterol/total cholesterol: 3.84/1.15/4.39/7.29 mmol/L), and hyperglycemia (blood glucose 7.07 mmol/L, HbA1c 7.2%). The patient also exhibited sinus tachycardia and elevated blood pressure, which was confirmed by 24-hour ambulatory blood pressure monitoring. The triglyceride-to-HDL cholesterol ratio was approximately 3.34, indicating insulin resistance and increased cardiovascular risk. Based on these findings, metabolic syndrome associated with obesity, type 2 diabetes mellitus, and hypertension was diagnosed. CONCLUSION:This case highlights the early manifestation of metabolic syndrome and illustrates its underlying pathophysiological mechanisms. Effective management requires early diagnosis, lifestyle modification, weight reduction, and individualized pharmacological therapy to improve metabolic control and reduce long-term cardiovascular risk.
Background/Aims: The aging process is a process of progressive decrease in cellular plasticity, with parallel changes in gene expression and epigenetic regulation. The microRNA-145 (miRNA-145) is reported to control transcription factors of pluripotency, but the connection between the microRNA and the DNA methylation process in human aging is not well understood. The aim of the study was to examine the relationship between miRNA-145 expression and transcriptional and epigenetic regulation of SOX2, KLF4, and MYC in various age groups. Methods: The quantitative real-time PCR was used to determine the expression levels of miRNA-145 and target genes. Methylation-specific PCR after bisulfite conversion was used to determine promoter methylation status. To assess the age-related changes in molecules, correlation and group-based analysis were conducted. Results: miRNA-145 expression showed a significant increase with age (p < 0.001) and was negatively correlated with SOX2 (r =-0.51), KLF4 (r =-0.55), and MYC (r =-0.47) expression. Simultaneously, there were age-dependent increases in the rates of promoter methylation of all three genes with a positive correlation with miRNA-145 levels. These results show that there are organized transcriptional and epigenetic changes in relation to aging. Conclusion: The findings are indicative of miRNA-145 being linked with age-related transcriptional and epigenetic modifications of crucial pluripotency genes. This consistent trend can be one of the causes of a drop in cellular plasticity that is experienced in human aging and underscores the promise of these molecular signatures as biomarkers of biological aging. (c) 2026 The Author(s). Published by Cell Physiol Biochem Press GmbH&Co. KG
BACKGROUND/AIMS:The Toll-like receptor 4 (TLR4) pathway plays a critical role in mediating inflammatory responses and regulates mitochondrial structural adaptations. Urolithin A (UA), a natural compound that induces mito- and autophagy, may improve mitochondrial homeostasis and health. This study examined the effects of UA on TLR4 signaling, mitochondrial dynamics, autophagy, and mitochondrial morphology after lipopolysaccharide (LPS) incubation in C2C12 myotubes. METHODS:C2C12 myotubes were incubated with LPS (1 µg/ml) and/or UA (50 µM). Protein expression related to TLR4 signaling, mitochondrial dynamics, and autophagy was assessed by Western blot, while mitochondrial morphology was evaluated using electron microscopy. RESULTS:LPS-induced inflammation activated downstream TLR4 signaling pathways and altered mitochondrial morphology, including reduced mitochondrial area and circularity. LPS also increased phosphorylation of the mitochondrial fission marker DRP1. UA prevented or attenuated these alterations induced by LPS. CONCLUSION:UA partially mitigates LPS-induced inflammation through modulation of TLR4 signaling and mitochondrial dynamics and may represent a promising therapeutic strategy for disorders associated with mitochondrial dysfunction and inflammation.
BACKGROUND/AIMS:Brominated flame retardants (BFRs) are widely used synthetic compounds with increasing environmental persistence and toxicological concern. This study investigated acetylcholinesterase (AChE) activity in human erythrocytes as a potential biomarker of BFR-induced cellular dysfunction. METHODS:AChE activity was measured spectrophotometrically using Ellman's method after exposure to selected compounds including tetrabromobisphenol A (TBBPA), tetrabromobisphenol S (TBBPS), and bromophenols. RESULTS:TBBPA and TBBPS significantly altered AChE activity, with increased enzyme activity observed at specific concentrations. Among bromophenols, significant effects were detected for 2,4,6-TBP and pentabromophenol. CONCLUSION:The biological effects of BFRs depend on chemical structure and concentration. While erythrocyte AChE responds to exposure, it is not a highly sensitive biosensor, as detectable changes occur alongside other cellular alterations.
BACKGROUND/AIMS:This study aimed to apply ultra-broadband micromechanical ultrasound (UMUS) for correction of myelosuppression caused by the cytotoxic effects of cyclophosphamide without limiting its antitumor efficacy. METHODS:The study included animals bearing transplanted Ehrlich carcinoma. Cyclophosphamide (CP) was administered once daily for three consecutive days starting on day 8 of tumor growth at a cumulative dose of 330 mg/kg per mouse. After completion of CP administration, a subset of animals was exposed to UMUS irradiation once daily for five days. Control groups included mice without tumors and tumor-bearing mice not exposed to CP or UMUS. Tumor growth kinetics were analyzed, and quantitative parameters of peripheral blood, bone marrow, and spleen were determined. RESULTS:The obtained data indicate that UMUS exposure does not reduce the antitumor efficacy of CP but is associated with enhanced recovery of the hematopoietic system and exerts a positive effect on bone marrow regeneration. This is confirmed by a statistically significant increase in the number of cells in specific bone marrow hematopoietic pools, including myelokaryocytes, blast cells, erythroid, lymphoid, and megakaryocytic cells. CONCLUSION:UMUS exposure was associated with accelerated recovery of multiple hematopoietic lineages in the bone marrow following cyclophosphamide-induced injury, without compromising antitumor efficacy.
BACKGROUND/AIMS:Rapid and cost-effective quantification of triglycerides remains a challenge in industrial food quality control, where conventional analytical methods are often time-consuming and resource-intensive. This study aimed to develop and characterize an amperometric biosensor based on lipase (LPS) as a biorecognition element for efficient triglyceride detection in vegetable oils. METHODS:Lipase was immobilized onto the electrode surface to catalyze the hydrolysis of glyceryl tributyrate, enabling electrochemical signal generation. Measurements were performed at an applied potential of -600 mV, with a detection time of 7 seconds. The enzymatic reaction rate was derived from the slope of the amperometric signal (mV/s), and calibration was conducted over a concentration range of 1-5 mM. RESULTS:The biosensor demonstrated a strong linear response (R² = 0.998) and high substrate affinity (KM = 0.13 mM). The system exhibited excellent repeatability and specificity, with minimal interference from other triglycerides. Long-term stability studies showed that the immobilized enzyme retained approximately 90% of its activity over 260 days and could be reused up to 27 times without significant performance loss. CONCLUSION:This biosensor represents a robust, rapid, and environmentally friendly analytical platform for triglyceride detection, offering a practical alternative to conventional techniques and strong potential for implementation in routine industrial quality control.
BACKGROUND/AIMS:We explored the fundamental feasibility, technical frame conditions and effectiveness of transcranial Tumor Destructive Mechanical Impulse (TMI) treatment of brain tumors. METHODS:The optimal treatment parameters (total energy required, energy flux density, shock wave frequency) and applicator positions were explored and defined with respect to the special conditions given by propagation of shock waves through the skull. RESULTS:As first clinical observations, we present the outcome of transcranial TMI treatment in two cases with brain metastases of malignant melanoma. In both cases, regression of metastases as well in the brain as in other sites was observed, in one of the cases with a complete tumor free recovery. CONCLUSION:In these cases, there were no identifiable signs of impairment of the surrounding brain tissue, and even indications of neuro-regenerative effects as described in the cited experimental literature, so that transcranial TMI treatment of brain tumors may be applied for further systematic investigation without any currently discernible risk for the intact surrounding brain tissue.
BACKGROUND/AIMS:Monitoring lactic acid is a critical parameter during the production of fermented foods, as it serves as an indicator of product quality and process control. The use of biosensors offers a simple and cost-effective approach for real-time monitoring in the production chain. METHODS:A biosensor based on lactate oxidase (LacOx) was developed to determine lactic acid content in fermented beverages (chicha and pozol). Different membranes were evaluated as immobilization supports, with particular focus on enzyme stability. The system operated at a potential of -0.5 mV, and the amperometric signal was recorded after 10 seconds. Sensor performance was assessed through calibration, sensitivity, and kinetic analysis. RESULTS:The enzymatic reaction rate was directly proportional to oxygen consumption (mg L⁻¹ O₂ s⁻¹), showing a linear detection range between 0.1 and 2 mM (R² = 0.9986). The sensor demonstrated good sensitivity at 0.1 mM and allowed membrane reuse up to 15 times. Enhanced stability was observed when using a cassava biopolymer as immobilization support. The Michaelis-Menten constant (KM = 340 µM) indicated a high affinity of the enzyme for the substrate. CONCLUSION:The developed biosensor provides a reliable, sensitive, and low-cost method for monitoring lactic acid in fermented beverages and represents a practical tool for quality control in the food industry.
BACKGROUND/AIMS:Nosocomial infections continue to be a serious problem in hospital settings, particularly in intensive care units (ICUs), where resistant germs can spread through the air. Air filters are often designed to capture particulate matter but not necessarily to kill live germs that remain on their surfaces. MATERIALS:To address these issues, polypropylene filter media were imbued with silver (AgNPs), zinc oxide (ZnO), and copper oxide (CuO) nanoparticles to impart antibacterial activity. Chemical reduction was used to create AgNPs, while sol-gel and precipitation methods were used to prepare ZnO and CuO nanoparticles. RESULTS:DLS and zeta potential measurements were utilized to calculate particle size distribution, polydispersity index, and surface charge. The filters were coated with nanoparticle suspensions by an alcohol-based dip-coating process, followed by heat treatment to solidify the deposited layers. Antibacterial activity was measured against Staphylococcus aureus (ATCC 25923) and Pseudomonas aeruginosa (ATCC 27853) using agar diffusion and direct-contact assays, and reusability was assessed over three cycles. Different variations were observed among the tested materials. Silver nanoparticles demonstrated the largest inhibition zones (18.2 mm for S. aureus and 16.7 mm for P. aeruginosa), followed by copper oxide nanoparticles, which showed moderate inhibition. Under identical conditions, zinc oxide exhibited the lowest inhibition. Over three reuse cycles, AgNP-coated filters retained the majority of the antibacterial activity with minimal decreases in inhibitory zone diameters. ZnO and CuO coatings, on the other hand, showed significantly higher decreases after many cleanings. Treatment of filters with nanoparticles significantly increased antibacterial efficacy compared to untreated controls (p < 0.05). CONCLUSION:In general, the addition of metal nanoparticles to polypropylene filter media appears to improve efficacy in lowering viable airborne bacterial loads. Silver-based coatings exhibited the highest resistance and effective antibacterial response among the studied materials, indicating their potential applicability in ventilation systems for the control of airborne infection within the limitations of the present three-cycle experimental framework.
BACKGROUND/AIMS:CUL5 acts as the scaffold protein of the E3 ligase complex in ubiquitin-dependent protein degradation pathways. Overexpression of CUL5 inhibits cellular proliferation, whereas inhibition of CUL5 expression induces proliferation and may contribute to clinical disorders including cancer. The effects of CUL5 depend on its post-translational modification by NEDD8 (neddylation), a process that represents a potential therapeutic target. This study explores the structure-function relationship between CUL5 and its neddylation status in vitro. METHODS:CUL5 was mutated at the putative neddylation site Lys (K) 724 to Arg (K724RCUL5) and at three potential neddylation sites K724, K727, and K728 (K724R/K727R/K728RCUL5). Because mutation of the PKA phosphorylation site Ser (S) 730 (S730ACUL5) was previously shown to induce neddylation, a K724R/S730ACUL5 mutant was also generated. Mutant and wild-type constructs were expressed in rat endothelial cells (RAMEC), T47D cancer cells, and COS-1 cells. Cellular proliferation, MAPK phosphorylation, ERα expression, and CUL5 neddylation status were analyzed, including treatment with the neddylation inhibitor MLN4924. RESULTS:Expression of wild-type CUL5 attenuated cell growth in RAMEC, T47D, and COS-1 cells. In contrast, expression of K724RCUL5 and K724R/S730ACUL5 mutants induced cellular growth, whereas the K724R/K727R/K728RCUL5 mutant had no significant effect on proliferation. In T47D cells, MAPK phosphorylation and estrogen receptor (ERα) expression were directly correlated with the neddylation status of CUL5. Western blot analysis of COS-1 cells treated with MLN4924 demonstrated that CUL5 remained neddylated in all mutant cell lines. CONCLUSION:These findings suggest that modification of CUL5 by NEDD8 may occur at multiple lysine residues and that multi-site neddylation may contribute to the diverse regulatory effects of CUL5 on cellular signaling and proliferation.
BACKGROUND/AIMS:Nanobiotechnology offers sustainable strategies to enhance plant resistance by activating innate immune responses. This study evaluates the effect of chitosan nanoparticles (CNPs) on transcriptional activation of defense-associated genes in Arabidopsis thaliana. METHODS:CNPs were produced via ionic gelation and sprayed on leaves of 4-week-old plants. Relative expression levels of PR1, PDF1.2, and WRKY70 were measured using qRT-PCR and the 2^-ΔΔCt method with normalization and statistical analysis. RESULTS:CNP treatment significantly increased PR1, PDF1.2, and WRKY70 expression compared to controls, indicating activation of salicylic acid and jasmonic acid/ethylene signaling pathways. WRKY70 showed the highest induction, suggesting a major regulatory role in coordinating defense signaling. CONCLUSION:Chitosan nanoparticles act as effective inducers of transcriptional activation of plant defense markers in Arabidopsis thaliana. Mechanisms including ROS signaling, calcium influx, MAPK cascades, and PTI/ETI responses warrant further investigation.
