Toxicology Mechanisms and Methods最新文献

Fixed-dose combinations (FDCs) offer therapeutic benefits like enhanced efficacy, reduced adverse effects, and better patient compliance, making them cost-effective. They are particularly effective in managing chronic obstructive pulmonary disease (COPD). Combining a long-acting muscarinic antagonist with a long-acting beta-agonist improves lung function, reduces the need for rescue bronchodilators, alleviates respiratory symptoms, and enhances the overall quality of life in COPD patients. This study evaluated the safety and tolerability of a novel FDC containing vilanterol, a selective β2-adrenoreceptor agonist, and glycopyrronium, an antimuscarinic agent, in Wistar rats. Vilanterol promotes bronchodilation, while glycopyrronium reduces bronchoconstriction. Repeated-dose toxicity testing at three dosage levels (6.25 + 12.5 mcg/kg/day, 12.5 + 25 mcg/kg/day, and 25 + 50 mcg/kg/day) through nose-only exposure showed that the FDC was well-tolerated, with no significant clinical signs of toxicity. Key parameters, including body weight, feed consumption, ophthalmic examination, clinical pathology, and bronchoalveolar lavage fluid analysis, showed no adverse effects. Minimal, non-dose-related microscopic lesions and normal alveolar macrophage responses were observed. The no-observed-adverse-effect level, based on actual concentration and duration of exposure, was established at 25 + 50 mcg/kg/day, indicating the FDC's safety and suitability for further development in COPD management.

Objectives: Databases specifying the nutrients and allergens present in multi-ingredient foods are required to explore the effect of food consumption on health outcomes accurately. The phytochemicals found in tree nuts have been associated with antioxidant, anti-inflammatory, antiproliferative, antiviral, chemo preventive and hypercholesterolaemic actions, all of which are known to affect the initiation and progression of several pathogenic processes. We have developed KNUTS-DB - a data-driven knowledge database for nuts - containing information on the chemical nutrients, molecular targets, pathways, disease associations, and clinically defined food allergen properties of peanuts and tree nuts.

Methods: The database includes data sets associated with extremely rich and diverse metadata on almonds, cashew, pecan, walnut, pistachio, peanut and walnut. Additionally, the database allows users to perform pathway and GO-Term enrichment analysis for user-defined chemical nutrients. The results of the analysis are presented using heatmaps and network plots.

Results: The database can be searched using options such as similarity search, interaction values, type of allergens, and specific nut types. KNUTS-DB offers researchers a unique platform to explore nuts' chemical composition and to investigate associations between nut composition and health outcomes - providing deeper insights into the molecular mechanisms.

Conclusions: KNUTS-DB, the first of its kind, is freely available to all users via https://allergypred.charite.de/KNutsDB/ without any login or registration.

Premature ovarian insufficiency/failure is a well-known long-term risk of chemotherapy including CDDP in women. Granulosa cells (GCs) are an essential ovarian cell type that promotes oocyte growth and is crucial for ovarian reproductive function. Hyperoside (HYP) is a flavonoid known for its beneficial pharmacological properties, including anti-inflammatory and antiapoptotic effects. Hence the current work aimed to evaluate the potential cytoprotective impact of HYP on CDDP-induced cytotoxicity in a human ovarian GCs cell line model via a wide range of assays including MTT, hormones secretion, ATP and mitochondrial membrane potential, reactive oxygen species, lipid peroxidation as well as antioxidant enzymes, Caspases, and Akt kinase activities. Forty-eight-hour exposure to 5-10µM CDDP resulted in reduction of GCs viability in a dose-dependent manner. HYP (40 µM) was found to ameliorate this CDDP -induced effect on GCs viability. CDDP in a concentration-dependent way, dramatically reduced cellular ATP, mitochondrial activities, cellular progesterone, and estradiol secretion. It also increased oxidative stress markers, cytochrome c levels, caspase -3.-8.-9, and Bax/Bcl2 ratio with decreased Akt kinase activity and its coding genes expression. These cytotoxic effects of CDDP on the treated GCs, were mitigated to varying degrees by HYP (40 µM). In conclusion, CDDP-induced cytotoxic effects on GCs seem to be the possible underlying cellular and molecular mechanisms of CDDP-induced ovarian insufficiency/failure. The study also demonstrated the therapeutic potential of HYP in mitigating CDDP-induced ovarian injury. Further studies are warranted to investigate the potential benefit of HYP as an adjuvant to CDDP treatment protocols to avoid adverse ovarian effects.

Mycotoxins perturb the gut microbiota performance. Bioactive compounds have been recently used as a new food strategy to diminish mycotoxins bioaccessibility and prevent their toxic effects on human and animal health. Male and female Wistar rats were exposed orally to twelve different diets containing aflatoxin B1 (AFB1) and/or ochratoxin A (OTA) with or without fermented whey (FW) and pumpkin (P) for 28 days. Fecal microbiota using 16S rRNA gene sequencing and subsequent metagenomics analysis were analyzed to study the effect of 28-day exposure through diet of contaminated and enriched feed. QIIME 2 microbiome analysis package (version 2024.5) was used to analyze the demultiplexed data. Mycotoxins-functional ingredients combination contributed more to microbial phylogenetic faith α-diversity rather than the functional ingredients alone, while the same combination reported a microbial α-diversity enhancement in comparison to the mycotoxins alone. Proteobacteria phylum was reduced in rat samples fed with contaminated diets (AFB1, OTA, and AFB1+OTA), while there was an increase-although not in all groups-when adding the functional ingredients. The main difference between the sexes was found in FW+AFB1+OTA group, with males (25%) showing higher % of Proteobacteria than females (1.86%). Phylogenetic diversity faith only focuses on microbial genetic (dis)similarity, not considering the biological function. Morganella morganii, a Proteobacteria found in some groups presents anticancer activity, but it is also related to inflammatory bowel disease and colorectal cancer. To sum up, both mycotoxins and functional ingredients trigger changes in the microbiota profile of Wistar rats in a sex-specific manner.

Hypoadrenocorticism is a serious condition in dogs that results from autoimmune adrenalitis and depletion of mineralocorticoids and glucocorticoids. Affected dogs respond well to glucocorticoid supplementation and treatment with the synthetic mineralocorticoid desoxycorticosterone pivalate (DOCP). DOCP injected once monthly resolves serum Na/K abnormalities and normalizes water balance, but therapy is expensive. Cost abatement involves prolongation of the 30-day dosage interval or decreasing the 2.2 mg/kg dosage. These approaches are not based on DOCP pharmacokinetics. A full assessment of the practicality of either approach would benefit from understanding drug pharmacokinetics, requiring measurement of DOCP and its esterase product desoxycorticosterone (DOC) in canine serum while avoiding toxic endpoints from overdosing. Mass spectrometric methods were developed including gas chromatography-tandem mass spectrometry of DOCP and DOC-methoxime trimethylsilyl derivatives, an approach sensitive to 2 ng/mL. Greater sensitivity was desired, so liquid chromatography-tandem mass spectrometry (LC-MS/MS) with ESI+ ionization was investigated. Supported liquid extraction was devised for serum with recoveries ∼100%. The LC-MS/MS method was validated for linearity, precision, accuracy and limits of detection (0.029 and 0.019 ng/mL for DOC and DOCP, respectively). A pilot experiment with DOCP-treated hypoadrenocorticism dogs over one-month revealed DOC baseline values as 0.183+/-0.090 ng/mL, which increased to the 1.0 - 2.2 ng/mL range. DOCP was not visible in any samples suggesting 100% conversion. Halving the dosage to 1.1 mg/kg still showed clear increases over the DOC baseline. MS fragmentation involved ring cleavages, dehydrations and double-charged fragments. The methodology was robust and suitable for studying DOC/DOCP pharmacokinetics in future studies of hypoadrenocorticism dogs.

Background: Benzo(a)pyrene (BaP) is an environmental pollutant linked to several cancers, including esophageal cancer (ESCA). Understanding its impact on gene expression and associated molecular pathways in ESCA is crucial for developing targeted therapies.

Methods: Using the TCGA-ESCA dataset, we identified differentially expressed genes (DEGs) related to BaP exposure. Enrichment analyses and protein-protein interaction (PPI) network construction were performed to explore the biological significance of these DEGs. Molecular docking studies assessed the interactions between BaP and core subnetwork genes. Survival analysis and immune cell infiltration analysis were conducted to evaluate the prognostic value of TGFBR2. Chemotherapy drug sensitivity was analyzed based on TGFBR2 expression levels.

Results: We identified 5757 DEGs in ESCA, with 33 genes linked to both BaP exposure and ESCA. Enrichment analyses revealed significant pathways, including p53 signaling and apoptosis. Key genes (ACTB, CDKN2A, TGFBR2) were verified for their differential expression. Molecular docking demonstrated strong BaP binding to several core proteins. High TGFBR2 expression correlated with better survival, enhanced immune infiltration, and altered sensitivity to chemotherapeutic agents.

Conclusion: Our study highlights the molecular mechanisms by which BaP influences ESCA, with TGFBR2 emerging as a potential prognostic marker and therapeutic target. These insights pave the way for personalized treatments in BaP-induced esophageal carcinogenesis.

This study investigates the protective effects of diacerein (DCN) against amiodarone (AMIO)-induced hepatotoxicity in a rat model. AMIO administration resulted in significant elevations of liver enzymes, ALT and AST, indicating hepatocellular membrane disruption and oxidative stress, as demonstrated by elevated levels of malondialdehyde (MDA) and decreased glutathione (GSH). Additionally, pro-inflammatory cytokines including TNF-α and IL-1β were expressed more when AMIO triggered the Toll-like receptor 4/nuclear factor kappa B/inflammasome 3 (TLR4/NF-κB/NLRP3) inflammatory pathway, along with elevated caspase-1 (CASP1) levels, which promoted apoptosis. In contrast, oral administration of DCN for two weeks effectively mitigated these effects by reducing liver enzyme levels and improving histopathological alterations. DCN also demonstrated anti-oxidant properties by decreasing MDA levels and increasing nuclear factor erythroid 2-related factor 2 (Nrf2) and GSH content. Furthermore, DCN downregulated the hepatic content of TLR4, NF-κB p65, NLRP3, CASP1, and pro-inflammatory cytokines, thereby inhibiting the activation of the inflammatory cascade. Moreover, DCN reduced protein expression of caspase 3. Those findings suggest that DCN exerts its hepatoprotective effects through its anti-oxidant activity, modulation of TLR4/NF-κB/NLRP3 inflammatory pathways, and reduction of apoptosis. These results provide new insights into potential therapeutic strategies for managing AMIO-induced hepatotoxicity, warranting further investigation into the underlying molecular mechanisms of DCN's protective effects.

Gliotoxin (GTX) is a potent mycotoxin that has been shown to induce neurotoxicity through the generation of oxidative stress and disruption of cellular signaling, leading to neuronal cell damage. The neurotoxic effects of GTX have been implicated in various neurodegenerative conditions, making the search for protective agents crucial. This study investigates the chemoprotective effects of acorn flour extract (ACFE) at different temperatures (20 °C, 60 °C, 80 °C, and 100 °C) on SH-SY5Y cells exposed to GTX using both pretreatment and simultaneous treatment strategies (direct treatment, pretreatment and simultaneous treatment). Cell viability was assessed using the MTT assay after 24 and 48 h of exposure. ACFE exhibited varying cytoprotective effects depending on the temperature and exposure conditions. Pre- treatment with 100 °C significantly increased cell viability by up to 51.6% at low GTX concentrations after 48 h; however, ACFE at 60 °C and 80 °C also demonstrated notable protective effects in pretreatment, suggesting a broader range of effective temperatures. Similarly, simultaneous treatment with ACFE (20 °C and 60 °C) enhanced cell viability by up to 124.7% at specific GTX concentrations. In general, higher extraction temperatures (80 °C and 100 °C) were associated with greater chemoprotective potential. These findings support the potential therapeutic application of ACFE in protecting against oxidative stress and neuronal damage, emphasizing the influence of extraction temperature and treatment timing on its efficacy. Further investigations are needed to explore the underlying molecular mechanisms involved in ACFE's protective effects.

3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (HMGR) inhibitors, commonly known as statins, are drugs frequently used in the treatment of hypercholesterolemia and hyperlipidemia. However, the current study has demonstrated that simvastatin induces neurotoxicity and is associated with cellular coenzyme Q₁₀ (CoQ₁₀) depletion. CoQ₁₀ has a significant role in the mitochondrial electron transport chain (ETC), in addition to being a fundamental lipid-soluble antioxidant. Depletion of CoQ₁₀ is frequently associated with impaired mitochondrial function and increased oxidative stress. The aim of this study was to investigate the potential mechanisms of simvastatin-induced neurotoxicity assessing mitochondrial function and evidence of oxidative stress in an in vitro SH-SY5Y human neuronal cell line. Fluorescence studies assessed via flow cytometry determined significant increases in intracellular and mitochondrial reactive oxygen species production following SH-SY5Y treatment with simvastatin compared to control cells. Additionally, spectrophotometric enzyme studies determined a significant (p < 0.0001) inhibition of ETC complex I and II-III activities which accompanied a significant decrease in neuronal CoQ₁₀ content (p < 0.005) and cell viability (p < 0.0001). The results of the present study have indicated evidence of mitochondrial dysfunction and increased oxidative stress, resulting in increased loss of neuronal viability following simvastatin treatment. Thus, these results demonstrate evidence of neurotoxicity associated with statin therapy.