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

Benchmark dose approach in investigating the relationship between blood metal levels and reproductive hormones: Data set from human study

The main objective of this research was to conduct a dose–response modeling between the internal dose of measured blood Cd, As, Hg, Ni, and Cr and hormonal response of serum testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH). The study included 207 male participants from subjects of 5 different cohorts (patients with prostate, testicular, and pancreatic cancer, patients suffering from various thyroid and metabolic disorders, as well as healthy volunteers), enrolled from January 2019 to May 2021 at the Clinical Centre of Serbia in Belgrade, Serbia. Benchmark dose–response modeling analysis was performed with the PROAST software version 70.1, showing the hormone levels as quantal data. The averaging technique was applied to compute the Benchmark dose (BMD) interval (BMDI), with benchmark response set at 10%. Dose-response relationships between metal/metalloid blood concentration and serum hormone levels were confirmed for all the investigated metals/metalloid and hormones. The narrowest BMDI was found for Cd-testosterone and Hg-LH pairs, indicative of high confidence in these estimates. Although further research is needed, the observed findings demonstrate that the BMD approach may prove to be significant in the dose–response modeling of human data

Akutna trovanja lekovima za terapiju kardiovaskularnih bolesti u Republici Srbiji

Acute poisonings by drugs used for cardiovascular diseases treatment are less frequent than poisonings by other drugs, but often lead to serious disorders in various organ systems and are characterized by relatively high mortality. When it comes to polydrug poisoning by drugs with a depressant effect on the cardiovascular system, hospital care of the patients is necessary. This research aimed to present data on acute poisonings by drugs used in the cardiovascular diseases treatment in the Republic of Serbia in the period 2010- 2018 based on the published Annual reports of the National Poison Control Center of the Military Medical Academy (1). Number of patients examined on suspicion of poisoning by drugs for cardiovascular diseases treatment, in the period 2010-2018, ranged from 4-6% of all examined due to acute drug poisoning per year, while number of hospitalizations due to drug poisoning ranged from 7 to 11% of all hospitalized patients. Of the total number of deaths resulting from drug poisoning, 27% are patients who died due to the poisoning by drugs used for cardiovascular diseases treatment. The most common causes of poisoning in the examined patients were beta blockers (45%), followed by Ca 2+ channel blockers (25%) and ACE inhibitors (21%), and these three groups of drugs are the most common causes in hospitalized patients as well. Lethal outcomes are most often the result of acute poisoning by Ca 2+ channel blockers (50%) and beta blockers (40%), mainly in combination with drugs used in psychiatric illnesses treatment, such as benzodiazepines, antiepileptics and antipsychotics.Akutna trovanja lekovima za lečenje kardiovaskularnih bolesti su ređa u odnosu na trovanja drugim lekovima, ali često dovode do ozbiljnih poremećaja u funkcionisanju različitih sistema organa i karakterišu se relativno visokim mortalitetom. Kada su u pitanju polimedikamentozna trovanja lekovima koji imaju depresorno dejstvo na kardiovaskularni sistem, neophodno je bolničko zbrinjavanje pacijenata. Cilj ovog rada je bio prikazati analizu podataka o akutnim trovanjima lekovima koji se koriste u terapiji kardiovaskularnih oboljenja u Republici Srbiji u periodu od 2010 do 2018. godine na osnovu publikovanih Godišnjaka Nacionalanog centra za kontrolu trovanja Vojnomedicinske akademije (1). Broj pacijenata koji su pregledani pod sumnjom na trovanje lekovima za lečenje kardiovaskularnih bolesti u periodu 2010-2018. godine kretao se od 4-6% svih pacijenata pregledanih usled akutnog trovanja lekovima godišnje, dok se broj pacijenata hospitalizovanih usled trovanja lekovima kretao 7-11% svih hospitalizovanih pacijenata. Od ukupnog broja letalnih ishoda koji su posledica trovanja lekovima, 27% čine pacijenti preminuli zbog trovanja lekovima za lečenje kardiovaskularnih bolesti. Najčešći uzročnici trovanja kod pregledanih pacijenata su beta blokatori (45%), zatim blokatori Ca 2+ kanala (25%) i ACE inhibitori (21%), a ove tri grupe lekova su najzastupljenije i kod hospitalizovanih pacijenata. Letalni ishodi su najčešće posledica akutnog trovanja blokatorima Ca 2+ kanala (50%) i beta blokatorima (40%), uglavnom u kombinaciji sa lekovima koji se koriste u terapiji psihijatrijskih oboljenja poput benzodiazepina, antiepileptika i antipsihotika.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Hepatotoksični potencijal smeše toluena, stirena i etanola: in silico toksikogenomička analiza

Organic solvents are still widely used in various industries and considered the most common chemicals associated with liver injury in workers. For research into the relationships between these chemicals and genes, interactions among chemicals, molecular pathways and biological processes, a significant place in toxicity testing has been taken by in silico methodologies. This study aims to provide evidence for the involvement of a selected mixture of organic solvents (toluene, styrene, ethanol) in liver disease development and show the potential of in silico toxicogenomic data-mining in determining possible mechanisms of mixture toxicity. The Comparative Toxicogenomics Database (CTD), GeneMania and ToppGene Suite were used for data-mining. The results showed that there were 17 genes connected with liver injury common for all the tested solvents. Co-expression (61.73%) was the most prominent interaction between the genes, while physical interactions were present at 14.56%, co-localization at 12.54% and interactions predicted by the server at 6.62%. Gene ontology analysis revealed biological processes affected by the investigated mixture (reactive oxygen species metabolic and biosynthetic process, response to oxidative stress, and response to organic cyclic compound). Oxidative stress response, antioxidant and oxidoreductase activity, vitamin B12 metabolism were noted as the key molecular pathways contributing to liver disease development. Our results emphasize the role of oxidative stress as one of the mechanisms of organic solvents' mixture toxicity and provide new insights into molecular mechanisms involved in hepatotoxicity.Organski rastvarači se još uvek široko koriste u raznim industrijama i smatraju se najčešćim hemikalijama povezanim sa oštećenjem jetre kod radnika. Za istraživanje odnosa između ovih hemikalija i gena, interakcija među hemikalijama, molekularnih puteva i bioloških procesa, značajno mjesto pripada i in silico metodologijama. Cilj ove studije je da pruži dokaze za povezanost odabrane smeše organskih rastvarača (toluen, stiren, etanol) u razvoju bolesti jetre, i da pokaže potencijal in silico toksikogenomičke analize podataka u određivanju mogućih mehanizama toksičnosti smješe. Za prikupljanje podataka korišteni su Comparative Toxicogenomics Database (CTD), GeneMania i ToppGene Suite. Rezultati ove analize su pokazali da postoji 17 gena povezanih s oštećenjem jetre zajedničkih za sva tri navedena rastvarača. Koekspresija (61,73%) bila je najistaknutija interakcija između gena, dok su fizičke interakcije bile prisutne sa 14,56%, kolokalizacije sa 12,54%, a interakcije predviđene od strane servera sa 6,62%. Analiza ontologije gena izdvojila je biološke procese na koje utiče ispitivana smeša (metabolički i biosintetski proces reaktivnih kiseonikovih vrsta, odgovor na oksidativni stres i odgovor na organska ciklična jedinjenja). Odgovor na oksidativni stres, aktivnost antioksidanata i oksidoreduktaze, i metabolizam vitamina B12 su navedeni kao ključni molekularni putevi koji dobrinose razvoju bolesti jetre. Rezultati ovog rada naglašavaju ulogu oksidativnog stresa kao jednog od mehanizama toksičnosti smeše organskih rastvarača i daju novi uvid u molekularne mehanizme uključene u hepatotoksičnost.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Put štetnog ishoda kao novi pristup u proceni toksičnosti u razvoju lekova

Adverse outcome pathways (AOPs) represent a tool in toxicology, introduced in 2010 by scientists from the US Environmental Protection Agency as a framework to support ecotoxicological research and risk assessment. In 2012, Organization for Economic Cooperation and Development initiated an international AOPs development program. Since then, AOPs have been promoted as useful tool in health risk assessment, development of Integrated Approaches to Testing and Assessment and for developing novel animal-free test methods (1,2). AOPs provide structured frameworks for collecting, organizing and evaluating existing toxicological knowledge on mechanistic pathways. AOPs describe biologically plausible chains of events, linking a molecular initiating event to key events at different levels of biological organization and, finally, to an adverse outcome. Not being stressor-specific, the ultimate utility of AOPs should be to predict adverse effects of any type of stressor, including chemicals, nanomaterials, pharmaceuticals, etc. with unknown toxic effects for which the toxicological mechanisms are known or can be tested. AOPs have been suggested as structured basis for predicting drug-induced liver injury (steatosis, fibrosis, cholestasis) and for developing in silico and in vitro methods for screening, as well as targeted methods for pre-clinical testing to assess liver toxicity, common reason for withdrawing pharmaceuticals from the market. AOPs have also been proposed as useful framework for method development and integration of in vitro data in personalized cancer therapy. In conclusion, AOPs concept plays an important role in the 21 st century toxicology paradigm supporting predictive toxicology with alternative assays and reduction of the need for animal use.Put štetnog ishoda (engl. adverse outcome pathway, AOP) predstavlja alatku u toksikologiji prvi put uvedenu 2010. godine od strane naučnika Američke agencije za zaštitu životne sredine sa ciljem podrške istraživanjima u ekotoksikologiji. Već 2012. godine Organizacija za ekonomsku saradnju i razvoj pokrenula je međunarodni program razvoja AOP. Od tada, AOP je promovisan kao koristan pristup u proceni rizika po zdravlje ljudi, razvoju novih metoda i integrisanih pristupa testiranju i evaluaciji štetnih efekata (1,2). AOP daje struktuirani okvir za prikupljanje, organizaciju i procenu postojećeg znanja o mehanističkim putevima u toksikologiji. AOP opisuje biološki verovatan lanac događaja povezujući inicijalni molekularni događaj, preko tzv. ključnih događaja na različitim nivoima biološke organizacije, sa štetnim efektom. AOP nije specifičan i kao takav, treba da posluži za predviđanje štetnih efekata različitih stresora, kao npr. hemikalija, nanomaterijala, lekova i drugih, sa nepoznatim štetnim efektom, ali za koje su poznati ili se mogu ispitati mehanizmi toksičnosti. Opisani su AOP u cilju predviđanja lekovima indukovanih oštećenja jetre (steatoza, fibroza, holestaza), kao i u cilju razvoja in silico i in vitro metoda za skrining i pretkliničko ispitivanje ovih efekata, koji su jedan od čestih razloga povlačenja lekova sa tržišta. AOP je predložen i kao koristan okvir za razvoj metoda i integraciju in vitro podataka u personalizovanu terapiju karcinoma. Konačno, AOP koncept igra važnu ulogu u toksikologiji 21. veka, koja podržava prediktivnu toksikologiju sa altentativnim metodama i smanjenjem potrebe za eksperimentalnim životinjama.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Povećani oksidacijski stres u obućarskih radnika izloženih mješavini niskih razina hlapljivih organskih spojeva

This study aimed to assess the redox status and trace metal levels in 49 shoe industry workers (11 men and 38 women) occupationally exposed to a mixture of volatile organic compounds (VOCs), which includes aliphatic hydrocarbons, aromatic hydrocarbons, ketones, esters, ethers, and carboxylic acids. All measured VOCs were below the permitted occupational exposure limits. The control group included 50 unexposed participants (25 men and 25 women). The following plasma parameters were analysed: superoxide anion (O2•-), advanced oxidation protein products (AOPP), total oxidative status (TOS), prooxidant-antioxidant balance (PAB), oxidative stress index (OSI), superoxide dismutase (SOD) and paraoxonase-1 (PON1) enzyme activity, total SH group content (SHG), and total antioxidant status (TAS). Trace metal levels (copper, zinc, iron, magnesium, and manganese) were analysed in whole blood. All oxidative stress and antioxidative defence parameters were higher in the exposed workers than controls, except for PON1 activity. Higher Fe, Mg, and Zn, and lower Cu were observed in the exposed vs control men, while the exposed women had higher Fe and lower Mg, Zn, and Cu than their controls. Our findings confirm that combined exposure to a mixture of VOCs, even at permitted levels, may result in additive or synergistic adverse health effects and related disorders. This raises concern about current risk assessments, which mainly rely on the effects of individual chemicals, and calls for risk assessment approaches that can explain combined exposure to multiple chemicals.Cilj istraživanja bio je procijeniti redoks status i koncentracije metala u tragovima kod 49 radnika (11 muškaraca i 38 žena) zaposlenih u industriji obuće, izloženih mješavini hlapljivih organskih spojeva (engl. volatile organic compounds, krat. VOC) koju su činili alifatski i aromatični ugljikovodici, ketoni, esteri, eteri i karboksilne kiseline. Sve izmjerene razine VOC bile su ispod dozvoljenih granica profesionalne izloženosti. U kontrolnoj je skupini bilo 50 neizloženih ispitanika (25 muškaraca i 25 žena). U plazmi obućarskih radnika analizirani su sljedeći parametri: superoksid anion radikal (O2 •- ), uznapredovali produkti oksidacije proteina (AOPP), totalni oksidacijski status (TOS), prooksidacijsko- antioksidacijski balans (PAB), indeks oksidacijskoga stresa (OSI), aktivnost superoksid-dismutaze (SOD), aktivnost enzima paraoksonaze-1 (PON1), ukupni sadržaj sulfhidrilnih grupa (SHG) i totalni antioksidacijski status (TAS). Koncentracije metala u tragovima (bakar, cink, željezo, magnezij i mangan) analizirane su u punoj krvi. U usporedbi s kontrolnom skupinom, svi parametri oksidacijskoga stresa i antioksidacijske obrane u izloženih radnika bili su povišeni, osim aktivnosti PON1. Povećana koncentracija Fe, Mg i Zn te smanjena koncentracija Cu uočene su u izloženih muškaraca u usporedbi s kontrolnima, a u žena je primijećena povišena koncentracija Fe, a niža koncentracija Mg, Zn i Cu u odnosu na kontrolnu skupinu. Naši nalazi potvrđuju da kombinirano izlaganje mješavini VOC, čak i pri dozvoljenim koncentracijama, može dovesti do aditivnih ili sinergističkih štetnih učinaka na zdravlje i popratnih poremećaja. Trenutna procjena rizika, koja se uglavnom oslanja na učinke pojedinačnih kemikalija, izaziva zabrinutost i upućuje na potrebu za novim pristupima koji će objasniti kombiniranu izloženost većem broju kemikalija

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