Objective: This study aimed to investigate the cytotoxic effects of the organophosphate pesticide malathion on triple-negative breast cancer (TNBC) cells and to examine its impact on superoxide dismutase 1 (SOD1) gene expression.Materials and Methods: Human MDA-MB-231 TNBC cells were treated with escalating concentrations of malathion. Cell viability was assessed using the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay, and the half-maximal inhibitory concentration (IC₅₀) was determined. Wound healing and matrigel transwell invasion assays were performed to evaluate cell migration and invasive capacity, respectively. Following exposure to the IC₅₀ concentration for 24 and 48 hours, SOD1 mRNA levels were measured via real-time quantitative polymerase chain reaction (RT-qPCR). Molecular docking analyses were performed using AutoDock and SeamDock to assess interactions between malathion and the human SOD1 protein (PDB ID: 2NAM).Results: Malathion reduced cell viability in a dose-dependent manner, with an IC₅₀ value of 17.79 μM after 48 hours of exposure. Treatment at this concentration significantly reduced both migratory activity (p<0.001) and invasive capacity (p<0.05). SOD1 mRNA expression decreased after 24 hours of exposure (0.6-fold relative to controls) and showed partial recovery at 48 hours (0.9-fold relative to controls). However, the difference between the two treatment groups was not statistically significant (p>0.05). Molecular docking predicted a binding affinity of −6.9 kcal/mol and revealed the formation of six hydrogen bonds between malathion and SOD1.Conclusion: Malathion exerted cytotoxic effects and impaired migration and invasion in MDA-MB-231 TNBC cells under cytotoxic exposure conditions. Although molecular docking predicted a potential interaction between malathion and SOD1, malathion did not significantly alter SOD1 transcription under the conditions examined. These findings should be considered preliminary and hypothesis-generating rather than definitive mechanistic evidence, and they warrant further investigation to clarify the molecular mechanisms underlying malathion-induced cellular responses.
Objectives: Bentazone is a widely used selective herbicide frequently detected in environmental water sources, raising concerns regarding its potential effects on non-target organisms. Although various pesticides have been associated with male reproductive toxicity, the effects of bentazone on testicular cells remain insufficiently characterized. This study aimed to assess the cytotoxic and apoptotic effects of bentazone in TM3 Leydig and TM4 Sertoli cells, with a particular focus on identifying potential cell type-specific responses.Materials and Methods: Cell viability using 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay was assessed using spectrophotometry, and intracellular reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), and apoptotic and necrotic cell populations were analyzed using flow cytometry.Results: Bentazone exposure resulted in concentration-dependent cytotoxicity, with TM3 cells showing greater sensitivity than TM4 cells (IC: 202.6 µM vs 464 µM). A dose-dependent increase in apoptosis was found in TM3 cells without significant changes in ROS production or MMP, indicating a ROS-independent response. In contrast, TM4 cells exhibited increased ROS production without accompanying MMP loss, along with a biphasic apoptotic pattern characterized by higher apoptosis at lower concentrations and attenuation at higher doses.Conclusion: Bentazone exhibits cytotoxic and cell type-specific apoptotic effects in TM3 and TM4 cells, likely involving multiple non-overlapping mechanisms beyond classical oxidative stress pathways. These findings contribute to filling an important gap in the literature regarding bentazone's reproductive toxicity potential and provide a basis for future hypothesis-driven investigations into the underlying molecular mechanisms.
Objective: Accurate assessment of biological effects in high-intensity focused ultrasound (HIFU) research is often limited by the lack of standardized calibration protocols for in vitro applications. This study established a metrology-guided calibration framework to evaluate the spatially confined thermal and biological effects of a Sonic Concepts H-102 HIFU transducer.Materials and Methods: Ultrasonic thermal doses were calculated using the Sapareto--Dewey model, and an automated 3D system was developed for precise temperature characterization. Biological effects of thermal doses (TD 60, TD 120, and TD 240 min) were investigated in HT-29 and HCT-116 colorectal cancer cell lines. Temperature elevation, cell viability (MTT assay), and apoptosis-related responses (TALI image cytometry and JC-1 assay) were compared between focal and adjacent wells.Results: The HIFU transducer achieved precise focal heating (50$degree$C - 52$degree$C) while avoiding significant thermal spread to adjacent regions (\>2 mm). HT-29 cells exhibited significant biological responses at focal sites with minimal effects in adjacent wells (p\<0.0001). HCT-116 cells demonstrated higher thermal sensitivity with less distinct focal confinement (p values ranged from \<0.001 to \>0.05), indicating that biological responses to indirect HIFU-related stress may vary according to intrinsic cell line characteristics.Conclusions: This metrology-guided platform provides a reliable, reproducible framework for standardized in vitro HIFU applications while revealing cell-line-specific differences in thermal sensitivity.
Several diagnostic, prognostic, and predictive biomarkers have been investigated in renal cell carcinoma (RCC), among which carbonic anhydrase IX (CAIX) is particularly relevant. Although CAIX is expressed in several RCC subtypes, its expression is particularly high in clear cell RCC (ccRCC), the most common RCC subtype and one with substantial metastatic potential. Given the tendency of ccRCC to form tumor emboli and disseminate, circulating tumor cells (CTCs) may serve as valuable prognostic and predictive markers. This review explores the potential of CAIX as a biomarker for CTC detection in ccRCC. Conventional CTC detection relies on epithelial markers, but these are often unreliable in ccRCC due to epithelial-mesenchymal transition (EMT). Alternative biomarker strategies, including CAIX-containing detection approaches, have shown promising preliminary performance and may help to address EMT-associated marker loss. However, their clinical utility requires validation in large prospective cohorts. Given the rarity and heterogeneity of CTCs, CAIX-containing enrichment strategies may contribute to prognostic assessment and longitudinal disease monitoring in patients with ccRCC. Additionally, this approach may provide deeper insights into tumor biology, potentially supporting more individualized treatment-monitoring strategies.
Objective: This study assessed whether human leukocyte antigen (HLA)-B signal peptide dimorphism at position −21 (HLA-B-21) Methionine/Threonine (M/T) and HLA-E polymorphisms are associated with the level of viremia in human immunodeficiency virus (HIV)-positive individuals from the Turkish population.Materials and Methods: The study cohort comprised 130 HIV--positive patients and 102 healthy controls. HLA typing was achieved through a next-generation sequencing (NGS)--based approach, and HLA-E alleles were determined by Sanger sequencing. HIV-1 level of viremia was measured using the COBAS Ampliprep/TaqMan system. Statistical evaluation included chi-square analysis, nonparametric techniques, and logistic regression modeling.Results: HLA-E\*01:01 and the homozygous E\*01:01 genotype were observed significantly more frequently in HIV--positive patients than in controls. No significant relationship was found between HLA-E alleles and viral load. The HLA-B\*51:01 allele was significantly more frequent in patients with a higher viral load, whereas HLA-B\*44:02 showed a similar trend without reaching statistical significance. In addition, the HLA-B-21 T/T genotype was significantly associated with higher viral load levels.Conclusion: Our findings indicate that the HLA-B-21 signal peptide polymorphism may play a role in modulating HIV replication, whereas HLA-E polymorphisms alone appear to have a limited impact. This study provides novel immunogenetic data from the Turkish population and highlights the potential of HLA-B-21 polymorphism as a genetic marker influencing HIV disease progression.
