Toxicity of phthalate and bisphenol a mixture and assessment of the protective effect of probiotic on rat and zebrafish models

Široka upotreba bis(2-etilheksil)ftalata (DEHP), dibutil ftalata (DBP) i bisfenola A (BPA) otvaraniz mogućnosti za istovremenu izloženost ljudi ovim toksičnim supstancama u svakodnevnomživotu. Cilj ove doktorske disertacije bio je da ispita toksičnost smeše DEHP, DBP i BPA u odnosuna pojedinačne supstance na eksperimentalnim modelima pacova i zebrice, razjasni mehanizmetoksičnosti smeše in silico toksikogenomičkom analizom podataka i utvrdi potencijalnu efikasnostprobiotika u umanjenju štetnih efekata ispitivane smeše kod pacova i zebrica. Nakon subakutneizloženosti pacova, smeša je proizvela značajne promene u telesnoj masi, unosu hrane i vode, nivouhormona, lipidnom profilu, pojedinim hematološkim i biohemijskim parametrima, patohistološkimnalazima tkiva, nivoima parametara oksidativnog stresa/antioksidativne zaštite i bioelementima.Neki od ovih efekata bili su prisutni samo u grupi tretiranoj smešom ili izraženiji u odnosu na grupetretirane pojedinačnim supstancama. Eksperiment na embrionima zebrica potvrdio je da jetoksičnost smeše izraženija u odnosu na toksičnost pojedinačnih supstanci, kao i da je efekat smešeprvenstveno diktiran efektima BPA. In silico analiza ukazala je na povezanost ispitivane smeše irazvoja različitih poremećaja pri čemu su kao dominantni molekularni mehanizmi izdvojenioksidativni stres i apoptoza. Pokazan je zaštitni efekat probiotika protiv štetnih efekata smeše,posebno u slučaju smanjenja telesne mase, unosa hrane, biohemijskih i hematoloških parametara,nivoa hormona, parametara redoks statusa i bioelemenata u plazmi, tkivu pankreasa, pluća, timusa,bubrega i testisa pacova, kao i protiv efekata na rast i razvoj, hepatotoksičnosti i letaliteta ispitivanesmeše kod embriona zebrica.The widespread use of bis(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) and bisphenolA (BPA) opens up a range of possibilities for simultaneous human exposure to these toxicsubstances. The aim of this doctoral dissertation was to examine DEHP/DBP/BPA mixture toxicityin relation to individual substances in rat and zebrafish experimental models, clarify themechanisms of toxicity by conducting in silico toxicogenomic analysis and determine the potentialefficacy of multicomponent probiotic in reducing the adverse effects of the tested mixture in ratsand zebrafish. After subacute exposure of rats, the mixture produced significant changes inbodyweight, food and water consumption, hormone levels, lipid profile, certain hematological andbiochemical parameters, pathohistological findings, redox status parameters and bioelements. Someeffects were present only or more prominent in the group treated with the mixture in relation to thegroups treated with individual substances. Zebrafish embryos experiment confirmed that mixturetoxicity was more pronounced than toxicity of individual substances, and that mixture effects wereprimarily dictated by BPA. In silico analysis indicated an association between the mixture anddevelopment of various disorders, with oxidative stress and apoptosis being the dominant molecularmechanisms. Probiotic protective effect against mixture harmful effects has been shown, especiallyin the case of bodyweight gain, food intake, biochemical and hematological parameters, hormonelevels, redox status parameters and bioelements in plasma, pancreatic, lung, thymus, kidney andtestis in rats, as well as against lethal, teratogenic and hepatotoxic effects in zebrafish embryos

Toxicity of phthalate and bisphenol a mixture and assessment of the protective effect of probiotic on rat and zebrafish models

Široka upotreba bis(2-etilheksil)ftalata (DEHP), dibutil ftalata (DBP) i bisfenola A (BPA) otvara niz mogućnosti za istovremenu izloženost ljudi ovim toksičnim supstancama u svakodnevnom životu. Cilj ove doktorske disertacije bio je da ispita toksičnost smeše DEHP, DBP i BPA u odnosu na pojedinačne supstance na eksperimentalnim modelima pacova i zebrice, razjasni mehanizme toksičnosti smeše in silico toksikogenomičkom analizom podataka i utvrdi potencijalnu efikasnost probiotika u umanjenju štetnih efekata ispitivane smeše kod pacova i zebrica. Nakon subakutne izloženosti pacova, smeša je proizvela značajne promene u telesnoj masi, unosu hrane i vode, nivou hormona, lipidnom profilu, pojedinim hematološkim i biohemijskim parametrima, patohistološkim nalazima tkiva, nivoima parametara oksidativnog stresa/antioksidativne zaštite i bioelementima. Neki od ovih efekata bili su prisutni samo u grupi tretiranoj smešom ili izraženiji u odnosu na grupe tretirane pojedinačnim supstancama. Eksperiment na embrionima zebrica potvrdio je da je toksičnost smeše izraženija u odnosu na toksičnost pojedinačnih supstanci, kao i da je efekat smeše prvenstveno diktiran efektima BPA. In silico analiza ukazala je na povezanost ispitivane smeše i razvoja različitih poremećaja pri čemu su kao dominantni molekularni mehanizmi izdvojeni oksidativni stres i apoptoza. Pokazan je zaštitni efekat probiotika protiv štetnih efekata smeše, posebno u slučaju smanjenja telesne mase, unosa hrane, biohemijskih i hematoloških parametara, nivoa hormona, parametara redoks statusa i bioelemenata u plazmi, tkivu pankreasa, pluća, timusa, bubrega i testisa pacova, kao i protiv efekata na rast i razvoj, hepatotoksičnosti i letaliteta ispitivane smeše kod embriona zebrica.The widespread use of bis(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) and bisphenol A (BPA) opens up a range of possibilities for simultaneous human exposure to these toxic substances. The aim of this doctoral dissertation was to examine DEHP/DBP/BPA mixture toxicity in relation to individual substances in rat and zebrafish experimental models, clarify the mechanisms of toxicity by conducting in silico toxicogenomic analysis and determine the potential efficacy of multicomponent probiotic in reducing the adverse effects of the tested mixture in rats and zebrafish. After subacute exposure of rats, the mixture produced significant changes in bodyweight, food and water consumption, hormone levels, lipid profile, certain hematological and biochemical parameters, pathohistological findings, redox status parameters and bioelements. Some effects were present only or more prominent in the group treated with the mixture in relation to the groups treated with individual substances. Zebrafish embryos experiment confirmed that mixture toxicity was more pronounced than toxicity of individual substances, and that mixture effects were primarily dictated by BPA. In silico analysis indicated an association between the mixture and development of various disorders, with oxidative stress and apoptosis being the dominant molecular mechanisms. Probiotic protective effect against mixture harmful effects has been shown, especially in the case of bodyweight gain, food intake, biochemical and hematological parameters, hormone levels, redox status parameters and bioelements in plasma, pancreatic, lung, thymus, kidney and testis in rats, as well as against lethal, teratogenic and hepatotoxic effects in zebrafish embryos

Dekodiranje molekulskog aspekta oštećenja oka prouzročenog ionizirajućim zračenjem pomoću rudarenja genomskih podataka

Even at low levels, exposure to ionising radiation can lead to eye damage. However, the underlying molecular mechanisms are not yet fully understood. We aimed to address this gap with a comprehensive in silico approach to the issue. For this purpose we relied on the Comparative Toxicogenomics Database (CTD), ToppGene Suite, Cytoscape, GeneMANIA, and Metascape to identify six key regulator genes associated with radiation-induced eye damage (ATM, CRYAB, SIRT1, TGFB1, TREX1, and YAP1), all of which have physical interactions. Some of the identified molecular functions revolve around DNA repair mechanisms, while others are involved in protein binding, enzymatic activities, metabolic processes, and post-translational protein modifications. The biological processes are mostly centred on response to DNA damage, the p53 signalling pathway in particular. We identified a significant role of several miRNAs, such as hsa-miR-183 and hsamiR-589, in the mechanisms behind ionising radiation-induced eye injuries. Our study offers a valuable method for gaining deeper insights into the adverse effects of radiation exposure.Izloženost ionizirajućem zračenju čak i pri niskim razinama može pridonijeti nastanku oštećenja oka. Međutim, osnovni molekulski mehanizmi i dalje nisu potpuno razjašnjeni. Cilj našega istraživanja bio je ispuniti tu nedostajuću kariku primjenom sveobuhvatnog in silico pristupa problemu. U tu svrhu, pomoću genomskih baza podataka, portala i poslužitelja (Comparative Toxicogenomics Database, ToppGene Suite portal, Cytoscape, GeneMANIA i Metascape), identificirano je šest ključnih regulacijskih gena koji su povezani s oštećenjem oka prouzročenog ionizirajućim zračenjem (ATM, CRYAB, SIRT1, TGFB1, TREX1 i YAP1) i koji su svi bili u fizičkoj interakciji. Neke od identificiranih molekulskih funkcija odnosile su se na mehanizme popravka oštećenja DNA, a druge su bile uključene u vezanje proteina, enzimsku aktivnost, metaboličke procese i posttranslacijske modifikacije proteina. Biološki procesi uglavnom su bili povezani s odgovorom na oštećenje DNA, pogotovo sa signalnim putem p53. Uočena je i značajna uloga nekoliko miRNA, poput hsa-miR-183 i hsa-miR-589, u mehanizmima povezanima s oštećenjem oka prouzročenog ionizirajućim zračenjem. Osim toga, u ovom je istraživanju opisana korisna metoda za ispitivanje štetnih učinaka izloženosti zračenju

Potential genomic biomarkers of obesity and its comorbidities for phthalates and bisphenol A mixture: In silico toxicogenomic approach

This in silico toxicogenomic study aims to explore the relationship between phthalates and bisphenol A (BPA) co-exposure and obesity, as well as its comorbid conditions, in order to construct a possible set of genomic biomarkers. The Comparative Toxicogenomics Database (CTD; http://ctd.mdibl.org) was used as the main data mining tool, along with GeneMania (https://genemania.org), ToppGene Suite (https://toppgene.cchmc.org) and DisGeNET (http://www. disgenet.org). Among the phthalates, bis(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) were chosen as the most frequently curated phthalates in CTD, which also share similar mechanisms of toxicity. DEHP, DBP and BPA interacted with 84, 90 and 194 obesity-related genes/proteins, involved in 67, 65 and 116 pathways, respectively. Among these, 53 genes/proteins and 42 pathways were common to all three substances. 31 genes/proteins had matching interactions for all three investigated substances, while more than half of these genes/proteins (56.49%) were in co-expression. 7 of the common genes/proteins (6 relevant to humans: CCL2, IL6, LPL, PPARG, SERPINE1, and TNF) were identified in all the investigated obesity comorbidities, while PPARG and LPL were most closely linked to obesity. These genes/proteins could serve as a target for further in vitro and in vivo studies of molecular mechanisms of DEHP, DBP and BPA mixture obesogenic properties. Analysis reported here should be applicable to any mixture of environmental chemicals and any disease present in CTD

PFAS Molecules: A Major Concern for the Human Health and the Environment

Per- and polyfluoroalkyl substances (PFAS) are a group of over 4700 heterogeneous compounds with amphipathic properties and exceptional stability to chemical and thermal degradation. The unique properties of PFAS compounds has been exploited for almost 60 years and has largely contributed to their wide applicability over a vast range of industrial, professional and non-professional uses. However, increasing evidence indicate that these compounds represent also a serious concern for both wildlife and human health as a result of their ubiquitous distribution, their extreme persistence and their bioaccumulative potential. In light of the adverse effects that have been already documented in biota and human populations or that might occur in absence of prompt interventions, the competent authorities in matter of health and environment protection, the industries as well as scientists are cooperating to identify the most appropriate regulatory measures, substitution plans and remediation technologies to mitigate PFAS impacts. In this review, starting from PFAS chemistry, uses and environmental fate, we summarize the current knowledge on PFAS occurrence in different environmental media and their effects on living organisms, with a particular emphasis on humans. Also, we describe present and provisional legislative measures in the European Union framework strategy to regulate PFAS manufacture, import and use as well as some of the most promising treatment technologies designed to remediate PFAS contamination in different environmental compartments

Trends in Anti-Tumor Effects of Pseudomonas aeruginosa Mannose-Sensitive-Hemagglutinin (PA-MSHA): An Overview of Positive and Negative Effects

Cancer is a leading cause of death worldwide, for which finding the optimal therapy remains an ongoing challenge. Drug resistance, toxic side effects, and a lack of specificity pose significant difficulties in traditional cancer treatments, leading to suboptimal clinical outcomes and high mortality rates among cancer patients. The need for alternative therapies is crucial, especially for those resistant to conventional methods like chemotherapy and radiotherapy or for patients where surgery is not possible. Over the past decade, a novel approach known as bacteria-mediated cancer therapy has emerged, offering potential solutions to the limitations of conventional treatments. An increasing number of in vitro and in vivo studies suggest that the subtype of highly virulent Pseudomonas aeruginosa bacterium called Pseudomonas aeruginosa mannose-sensitive-hemagglutinin (PA-MSHA) can successfully inhibit the progression of various cancer types, such as breast, lung, and bladder cancer, as well as hepatocellular carcinoma. PA-MSHA inhibits the growth and proliferation of tumor cells and induces their apoptosis. Proposed mechanisms of action include cell-cycle arrest and activation of pro-apoptotic pathways regulated by caspase-9 and caspase-3. Moreover, clinical studies have shown that PA-MSHA improved the effectiveness of chemotherapy and promoted the activation of the immune response in cancer patients without causing severe side effects. Reported adverse reactions were fever, skin irritation, and pain, attributed to the overactivation of the immune response. This review aims to summarize the current knowledge obtained from in vitro, in vivo, and clinical studies available at PubMed, Google Scholar, and ClinicalTrials.gov regarding the use of PA-MSHA in cancer treatment in order to further elucidate its pharmacological and toxicological properties

Conducting bioinformatics analysis to predict sulforaphane-triggered adverse outcome pathways in healthy human cells

Sulforaphane (SFN) is a naturally occurring molecule present in plants from Brassica family. It becomes bioactive after hydrolytic reaction mediated by myrosinase or human gastrointestinal microbiota. Sulforaphane gained scientific popularity due to its antioxidant and anti-cancer properties. However, its toxicity profile and potential to cause adverse effects remain largely unidentified. Thus, this study aimed to generate SFN-triggered adverse outcome pathway (AOP) by looking at the relationship between SFN-chemical structure and its toxicity, as well as SFN-gene interactions. Quantitative structure-activity relationship (QSAR) analysis identified 2 toxophores (Derek Nexus software) that have the potential to cause chromosomal damage and skin sensitization in mammals or mutagenicity in bacteria. Data extracted from Comparative Toxicogenomics Database (CTD) linked SFN with previously proposed outcomes via gene interactions. The total of 11 and 146 genes connected SFN with chromosomal damage and skin diseases, respectively. However, network analysis (NetworkAnalyst tool) revealed that these genes function in wider networks containing 490 and 1986 nodes, respectively. The over-representation analysis (ExpressAnalyst tool) pointed out crucial biological pathways regulated by SFN-interfering genes. These pathways are uploaded to AOP-helpFinder tool which found the 2321 connections between 19 enriched pathways and SFN which were further considered as key events. Two major, interconnected AOPs were generated: first starting from disruption of biological pathways involved in cell cycle and cell proliferation leading to increased apoptosis, and the second one connecting activated immune system signaling pathways to inflammation and apoptosis. In both cases, chromosomal damage and/or skin diseases such as dermatitis or psoriasis appear as adverse outcomes

The role of toxic metals and metalloids in nrf2 signaling

Nuclear factor erythroid 2-related factor 2 (Nrf2), an emerging regulator of cellular resis-tance to oxidants, serves as one of the key defensive factors against a range of pathological processessuch as oxidative damage, carcinogenesis, as well as various harmful chemicals, including metals.An increase in human exposure to toxic metals via air, food, and water has been recently observed,which is mainly due to anthropogenic activities. The relationship between environmental exposureto heavy metals, particularly cadmium (Cd), lead (Pb), mercury (Hg), and nickel (Ni), as well asmetaloid arsenic (As), and transition metal chromium (Cr), and the development of various humandiseases has been extensively investigated. Their ability to induce reactive oxygen species (ROS)production through direct and indirect actions and cause oxidative stress has been documentedin various organs. Taking into account that Nrf2 signaling represents an important pathway inmaintaining antioxidant balance, recent research indicates that it can play a dual role depending onthe specific biological context. On one side, Nrf2 represents a potential crucial protective mechanismin metal-induced toxicity, but on the other hand, it can also be a trigger of metal-induced carcinogen-esis under conditions of prolonged exposure and continuous activation. Thus, this review aims tosummarize the state-of-the-art knowledge regarding the functional interrelation between the toxicmetals and Nrf2 signaling

Toxic Effects of the Mixture of Phthalates and Bisphenol A-Subacute Oral Toxicity Study in Wistar Rats

Phthalates and bisphenol A, classified as endocrine disruptors, have weak estrogenic, anti-androgenic properties, and aect thyroid hormone regulation. The aim of this study on male rats was to compare the subacute toxic effects of low doses of single compounds (bis (2 –ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP), and bisphenol A (BPA)) with the effects of their mixture through dierent biochemical, hormonal, and hematological parameters. Rats were divided into five experimental groups: Control (corn oil), DEHP (50 mg/kg b.w./day), DBP (50 mg/kg b.w./day), BPA (25 mg/kg b.w./day), and MIX (50 mg/kg b.w./day DEHP + 50 mg/kg b.w/day DBP + 25 mg/kg b.w./day BPA). Animals were sacrificed after 28 days of oral treatment and blood was collected for further analysis. The results demonstrated that the mixture produced significant changes in lipid profile, liver-related biochemical parameters, and glucose level. Furthermore, the opposite effects of single substances on the thyroxine level have been shown in comparison with the mixture, as well as a more pronounced effect of the mixture on testosterone level. This study contributes to the body of knowledge on the toxicology of mixtures and gives one more evidence of the paramount importance of mixture toxicity studies, especially in assessing the endocrine disruptive effects of chemicals