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

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

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

In silico analiza utjecaja toksičnih metala na komplikacije bolesti COVID-19: molekularni uvidi

COVID-19 can cause a range of complications, including cardiovascular, renal, and/or respiratory insufficiencies, yet little is known of its potential effects in persons exposed to toxic metals. The aim of this study was to answer this question with in silico toxicogenomic methods that can provide molecular insights into COVID-19 complications owed to exposure to arsenic, cadmium, lead, mercury, nickel, and chromium. For this purpose we relied on the Comparative Toxicogenomic Database (CTD), GeneMANIA, and ToppGene Suite portal and identified a set of five common genes (IL1B, CXCL8, IL6, IL10, TNF) for the six metals and COVID-19, all of which code for pro-inflammatory and anti-inflammatory cytokines. The list was expanded with additional 20 related genes. Physical interactions are the most common between the genes affected by the six metals (77.64 %), while the dominant interaction between the genes affected by each metal separately is co-expression (As 56.35 %, Cd 64.07 %, Pb 71.5 %, Hg 81.91 %, Ni 64.28 %, Cr 88.51 %). Biological processes, molecular functions, and pathways in which these 25 genes participate are closely related to cytokines and cytokine storm implicated in the development of COVID-19 complications. In other words, our findings confirm that exposure to toxic metals, alone or in combinations, might escalate COVID-19 severity.COVID-19 može izazvati niz komplikacija, uključujući kardiovaskularnu, bubrežnu i/ili respiratornu insuficijenciju, ali se malo zna o njegovim potencijalnim učincima u osoba koje su izložene toksičnim metalima. Cilj ovog istraživanja bio je odgovoriti na to pitanje pomoću in silico toksikogenomske metode, koja može pružiti molekularni uvid u komplikacije bolesti COVID-19 uslijed izloženosti arsenu, kadmiju, olovu, živi, niklu i kromu. U tu su svrhu korišteni Komparativna toksikogenomska baza podataka (CTD), GeneMANIA i ToppGene Suite portal te je identificirana skupina od pet zajedničkih gena (IL1B, CXCL8, IL6, IL10, TNF) za šest metala i COVID-19, koji svi kodiraju proinflamatorne i antiinflamatorne citokine. Lista je proširena s dodatnih 20 srodnih gena. Fizičke interakcije dominirale su između gena na koje utječe kombinacija ispitivanih metala (77,64 %), a koekspresija je dominantna interakcija između gena na koje djeluju pojedinačni metali (As 56,35 %, Cd 64,07 %, Pb 71,5 %, Hg 81,91 %, Ni 64,28 %, Cr 88,51 %). Biološki procesi, molekulske funkcije i putovi u kojima sudjeluje tih 25 gena blisko su povezani s citokinima i citokinskom olujom, koja je uključena u razvoj komplikacija bolesti COVID-19. Drugim riječima, ovi rezultati potvrđuju da izloženost toksičnim metalima, bilo pojedinačno ili u kombinaciji, može dovesti do razvoja težih oblika bolesti COVID-19

Zajednički utjecaj ključnih onečišćivača zraka na težinu COVID-a 19 – predviđanje zasnovano na analizi toksikogenomičkih podataka

Considering that some researchers point to a possible influence of air pollution on COVID-19 transmission, severity, and death rate, the aim of our in silico study was to determine the relationship between the key air pollutants [sulphur dioxide (SO2), carbon monoxide (CO), particulate matter (PMx), nitrogen dioxide (NO2), and ozone (O3)] and COVID-19 complications using the publicly available toxicogenomic analytical and prediction tools: (i) Comparative Toxicogenomic Database (CTD) to identify genes common to air pollutants and COVID-19 complications; (ii) GeneMANIA to construct a network of these common and related genes; (iii) ToppGene Suite to extract the most important biological processes and molecular pathways; and (iv) DisGeNET to search for the top gene-disease pairs. SO2, CO, PMx, NO2, and O3 interacted with 6, 6, 18, 9, and 12 COVID-19-related genes, respectively. Four of these are common for all pollutants (IL10, IL6, IL1B, and TNF) and participate in most (77.64 %) physical interactions. Further analysis pointed to cytokine binding and cytokine-mediated signalling pathway as the most important molecular function and biological process, respectively. Other molecular functions and biologica processes are mostly related to cytokine activity and inflammation, which might be connected to the cytokine storm and resulting COVID-19 complications. The final step singled out the link between the CEBPA gene and acute myelocytic leukaemia and between TNFRSF1A and TNF receptor-associated periodic fever syndrome. This indicates possible complications in COVID-19 patients suffering from these diseases, especially those living in urban areas with poor air quality.COVID-19 (engl. coronavirus disease 2019) respiratorna je bolest prouzročena infekcijom SARS-CoV-2 virusom (engl. severe acute respiratory syndrome coronavirus 2). Pretpostavlja se da postoji utjecaj atmosferskih čimbenika, uključujući i onečišćenje zraka, na prenošenje koronavirusa, njegovu težinu i stopu smrtnosti. Stoga je cilj ovoga in silico istraživanja bio utvrditi odnos između ključnih onečišćivača zraka [sumporova dioksida (SO2), ugljikova monoksida (CO), lebdećih čestica (PMx), dušikova dioksida (NO2), ozona (O3)] i komplikacija COVID-a 19 korištenjem: (i) komparativne toksikogenomičke baze podataka (engl. Comparative Toxicogenomic Database, CTD) za dobivanje gena, međusobno povezanih s onečišćivačima zraka i komplikacijama COVID-a 19, (ii) GeneMANIA servera za konstruiranje mreže između dobivenih I srodnih gena, (iii) ToppGene Suite za izdvajanje najvažnijih bioloških procesa/molekularnih puteva i (iv) DisGeNET baze podataka za traženje najvažnijih parova gen-bolest. Za SO2, CO, PMx, NO2 odnosno O3 utvrđena je interakcija sa 6, 6, 18, 9, odnosno 12 gena povezanih s komplikacijama COVID-a 19. Četiri su zajednička (IL10, IL6, IL1B i TNF) i u najvećem postotku (77,64 %) sudjeluju u fizičkim interakcijama. Vezivanje citokina i signalni put posredovan citokinima izdvojeni su kao najvažnija molekularna funkcija i biološki proces. Druge molekularne funkcije i biološki procesi uglavnom su bili povezani s aktivnošću citokina i upalom, što bi se moglo dovesti u vezu s citokinskom olujom i posljedičnim komplikacijama COVID-a 19. Utvrđena je veza između različitih bolesti i ispitivanih gena, posebice između CEBPA i akutne mijelogene leukemije (AML) te između TNFRSF1A i sindroma periodične vrućice povezanoga s TNF receptorom. To upozorava na moguće komplikacije u osoba zaraženih koronavirusom koje boluju od tih bolesti, poglavito kada su dodatno potaknute poremećajem funkcije spomenutih gena

Predviđanje štetnih efekata sulforafana in silico ispitivanjem njegovog ciljanog dejstva na gene, protein‐protein interakcije i klase molekula

Alternative form of cancer treatment includes targeting natural compounds such as sulfur-rich dietary phytochemical sulforaphane (SFN). However, data on SFN safety, interactions on the protein level and target of SFN in human organism are limited (1). The aim of this study was to elucidate the target interactions of SFN in human body in order to rationalize possible side-effects and predict off-targets by using in silico approach. STITCH database (http://stitch.embl.de) was used to obtain the information about chemical–protein interactions, while Metascape (https://metascape.org/) highlighted protein-protein interaction enrichment (PPIE). SwissTargetPrediction (http://www.swisstargetprediction.ch/) indicated the target molecule classes of SFN in human. Human genes that had the strongest interaction with SFN were NQO1, NFE2L2, CASP3, HSP90AA1, MAPK14, HDAC6, HPGDS, KEAP1, GSTA1 and GSTM1. PPIE analysis singled out fluid shear stress and atherosclerosis, NRF2 pathway and chemical carcinogenesis - reactive oxygen species (ROS) as the most significant interactions. The most represented class of SFN targeted molecules in human organism were enzymes (26.7%). Epidermal growth factor receptor erbB1, macrophage migration inhibitory factor, nitric oxide synthase (inducible) showed the highest probability target rate. In our previous study (2), we pointed out that the genome of cancer patients could affect SFN safety. The current study provides a set of target genes, emphasizes the importance of oxidative stress in the suggested genetic interactions and predicts classes of target molecules, which should further be examined.Alternativni oblik lečenja raka uključuje upotrebu prirodnih jedinjenja kao što je fitohemikalija bogata sumporom, sulforafan (SFN). Međutim, podaci o interakcijama SFN na nivou proteina i ciljnih mesta dejstva SFN u ljudskom organizmu su ograničeni (1). Cilj ove studije bio je da se ukaže na ciljne interakcije SFN kod ljudi kako bi se racionalizovali mogući neželjeni efekti i predvidela nova ciljna mesta toksičnosti korišćenjem in silico pristupa. STITCH baza podataka (http://stitch.embl.de) korišć ena je za dobijanje informacija o interakcijama između hemikalija i proteina, dok je Metascape (https://metascape.org/) izdvojio protein-protein interakcije (PPIE). SwissTargetPrediction (http://vvv.svisstargetprediction.ch/) ukazao je na ciljana mesta dejstva SFN kod ljudi. Izdvojeni su geni koji kod ljudi imaju najjaču interakciju sa SFN: NQO1, NFE2L2, CASP3, HSP90AA1, MAPK14, HDAC6, HPGDS, KEAP1, GSTA1, GSTM1. Ateroskleroza, NRF2 signalni put i hemijska karcinogeneza - reaktivne vrste kiseonika (ROS) označeni su kao najznačajnije protein-protein interakcije. Najzastupljenija klasa SFN ciljanih molekula u ljudskom organizmu bili su enzimi (26,7%). Receptor epidermalnog faktora rasta erbB1, faktor inhibitora migracije makrofaga i sintaza azot oksida (inducibilna) pokazali su najveć u stopu verovatnoć e ciljnog mesta dejstva. U našoj prethodnoj studiji (2) istakli smo da bi genom pacijenata obolelih od raka mogao uticati na bezbednost primene SFN. Međutim, ova studija daje dodatni set ciljnih gena i naglašava važnost oksidativnog stresa u predloženim interakcijama između gena, kao i predviđenim klasama ciljnih molekula na koje deluje SFN i koje bi trebalo dalje ispitati.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Subakutna toksičnost smeše ftalata i bisfenola a: procena protektivnog dejstva probiotika

The aim of this study was to: (1) examine subacute toxicity of bis (2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) and bisphenol A (BPA) mixture in rats and compare it with individual substance effects; (2) investigate the mechanisms of toxicity on critical target organ, testes (in vivo/in silico study); (3) examine multibiotic ability to mitigate mixture toxicity. Male rats were divided into groups (n = 6): (1) Control; (2) P (probiotic (8.78*10 8 CFU/kg bw/day)); (3) DEHP (50 mg/kg bw/day); (4) DBP (50 mg/kg bw/day); (5) BPA (25 mg/kg bw/day); and (6) MIX (50 mg/kg bw/day DEHP + 50 mg/kg bw/day DBP + 25 mg/kg bw/day BPA) (7) MIX + P. In silico toxicogenomic analysis was performed by Comparative Toxicogenomic Database (CTD), Citoscape software and ToppGene Suite portal. MIX led to significant changes in tissue structure (liver, kidney, spleen and testis), biochemical parameters and testosterone level of rats compared to the control group and individual substances. In silico analysis revealed 20 genes associated with DEHP/DBP/BPA and male reproductive system disorders, while the most probable mechanisms included metabolism, aryl hydrocarbon receptor pathway, apoptosis, and oxidative stress. Oxidative stress was confirmed in vivo, and changes in parameters in testicular tissue homogenates were most pronounced or present only in MIX group. Probiotic annulled/mitigated changes in biochemical, hematological, and oxidative stress parameters, relative liver mass, food consumption, and organ pathohistology. Hence, the obtained results indicate the possibility of future considerations of protective probiotic effects against phthalates and BPA mixture toxicity.Cilj ove studije bio je: (1) ispitati subakutnu toksičnost smeše bis (2-etilheksil) ftalata (DEHP), dibutil ftalata (DBP), bisfenola A (BPA) na pacovima i uporediti je sa efektima pojedinačnih supstanci; (2) istražiti mehanizme toksičnosti na kritičnom ciljnom organu, testisima (in vivo/in silico studija); (3) ispitati sposobnost višekomponentnog probiotika da ublaži toksičnost smeše. Mužjaci pacova podeljeni su u grupe (n = 6): (1) Kontrola; (2) P (probiotik (8.78 * 108 CFU/kg/day)); (3) DEHP (50 mg/kg t.m/dan); (4) DBP (50 mg/kg t.m/dan); (5) BPA (25 mg/kg t.m/dan); i (6) MIX (50 mg/kg t.m/dan DEHP + 50 mg/kg t.m/dan DBP + 25 mg/kg t.m/dan BPA) (7) MIX + P. In silico toksikogenomička analiza sprovedena je Komparativnom toksikogenomičkom bazom podataka (CTD), Citoscape softverom i ToppGene Suite portalom. Smeša je dovela do značajnih promena u strukturi tkiva (jetra, bubreg, slezina i testis), biohemijskim parametrima i nivou testosterona pacova u poređenju sa kontrolnom grupom i pojedinačnim supstancama. In silico analizom otkriveno je 20 gena povezanih sa smešom DEHP/DBP/BPA i poremećajima muškog reproduktivnog sistema, dok su najverovatniji mehanizmi uključili metabolizam, molekularni put povezan sa receptorom za aromatične ugljovodinike, apoptozu i oksidativni stres. Oksidativni stres potvrđen je in vivo, a promene u parametrima u homogenatu tkiva testisa bile su najizraženije ili prisutne samo u MIX grupi. Probiotik je otklonio ili ublažio promene u biohemijskim, hematološkim, i parametrima oksidativnog stresa, kao i relativnoj masi jetre, unosu hrane i patohistološkim nalazima izazvanim smešom ispitivanih supstanci. Dobijeni rezultati ukazuju na mogućnost budućih razmatranja protektivnih efekata probiotika protiv toksičnosti smeše ftalata i BPA.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Subacute Exposure to Low Pb Doses Promotes Oxidative Stress in the Kidneys and Copper Disturbances in the Liver of Male Rats

Recent data indicate that lead (Pb) can induce adverse effects even at low exposure levels. Moreover, the corresponding mechanisms of low Pb toxicity have not been well identified. In the liver and the kidneys, Pb was found to induce various toxic mechanisms leading to organ physiological disruption. Therefore, the purpose of the study was to simulate low-dose Pb exposure in an animal model with the aim of assessing oxidative status and essential element levels as the main mechanism of Pb toxicity in the liver and kidneys. Furthermore, dose–response modelling was performed in order to determine the benchmark dose (BMD). Forty-two male Wistar rats were divided into seven groups: one control group, and six groups treated for 28 days with 0.1, 0.5, 1, 3, 7, and 15 mg Pb/kg b.w./day, respectively. Oxidative status parameters (superoxide dismutase activity (SOD), superoxide anion radical (O2−), malondialdehyde (MDA), total sulfhydryl groups (SHG), and advanced oxidation protein products (AOPP)) and Pb, copper (Cu), zinc (Zn), manganese (Mn), and iron (Fe) levels were measured. Lowering Cu levels (BMD: 2.7 ng/kg b.w./day), raising AOPP levels (BMD: 0.25 µg/kg b.w./day) in the liver, and inhibiting SOD (BMD: 1.3 ng/kg b.w./day) in the kidneys appear to be the main mechanisms of Pb toxicity. The lowest BMD was derived for a decrease in Cu levels in liver, indicating that this effect is the most sensitive