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

Nivoi kadmijuma u humanom tkivu dojke i nivoi estradiola u serumu: Postoji li veza?

Cadmium (Cd), one of the most abundant environmental pollutants, is considered to have endocrine disrupting properties. However, data on the dose-response relationship between Cd dose and levels of hormones have been insufficiently studied, especially in human data sets. Thus, the aim of this study was to determine the possibility of analyzing data obtained from a case- control study in female patients with benign/malignant breast tumors, using the Benchmark dose (BMD) concept. The collected data on Cd levels in breast tissue and estrogen serum levels were processed in PROAST software using different variables. The dose-response relationship between the internal dose of Cd and estradiol levels in the serum was investigated and BMD intervals were calculated. The dose-response relationship between the Cd concentration in breast tissue and the estradiol serum level was shown, indicating lower estradiol serum levels as a consequence of higher Cd concentrations in breast tissue. As one of the few studies analyzing human data using the BMD approach, these findings could have a pivotal role in dose response analysis of data collected from human studies.Kadmijum (Cd), jedan od najzastupljenijih zagađivača životne sredine, dokazan je endokrini ometač. Međutim, podaci o postojanju odnosa između doze Cd i odgovora-nivoa hormona nisu dovoljno istraženi, posebno podaci sakupljeni iz studija na ljudima. Stoga je cilj ove studije bio da se utvrdi mogućnost analize podataka dobijenih iz studije slučaja-kontrole kod pacijentkinja sa benignim/malignim tumorom dojke, primenom koncepta Benčmark doze (BMD). Prikupljeni podaci o nivoima Cd u tkivu dojke i serumskim nivoima estrogena obrađeni su u PROAST softveru uz korišćenje različitih varijabli. Ispitivan je odnos doza-odgovor između unutrašnje doze Cd (koncentracije u tkivu dojke) i estradiola u serumu i izračunati BMD intervali. Utvrđeno je postojanje odnosa između koncentracije Cd u tkivu dojke i nivoa estradiola u serumu koje ukazuje na niže nivoe estradiola u serumu kao posledica veće koncentracije Cd u tkivu dojke. Kao jedno od retkih istraživanja ovog tipa, dobijeni rezultati mogli bi predstavljati početak otkrivanja mogućnosti analize podataka prikupljenih u studijama na ljudima primenom BMD pristupa

Redoks status i razine bioelemenata u mozgu štakora akutno izloženih smjesi kadmija i olova

Most Pb and Cd neurotoxicity studies investigate exposure to either of the toxic metals alone, while data on co-exposure are scarce. The aim of our study was to fill that gap by investigating acute combined effects of Pb and Cd on redox and essential metal status in the brain of Wistar rats. Animals were randomised in four groups of six to eight rats, which received 15 or 30 mg/kg of Cd, 150 mg/kg of Pb, or 150 mg/kg of Pb + 15 mg/kg of Cd by gavage. The fifth, control, group received distilled water only. Co-treatment with Pb and Cd induced significant increase in malondialdehyde (MDA) and thiobarbituric acid-reactive substances (TBARS) compared to control and groups receiving either metal alone. This is of special importance, as MDA presence in the brain has been implicated in many neurodegenerative disorders. The groups did not significantly differ in Zn, Cu, Mn, and Fe brain levels. Our findings highlight the importance of metal mixture studies. Neurotoxicity assessments of single chemicals do not provide a real insight into exposure to mixtures in real life. Further research should look into interactions between these metals to reveal complex molecular mechanisms of their neurotoxicity.Dosadašnji podatci o neurotoksičnosti olova (Pb) i kadmija (Cd) uglavnom su opisani nakon izloženosti pojedinačnim metalima, a podatci o neurotoksičnom djelovanju smjese tih dvaju metala prilično su oskudni. Cilj ovoga rada bio je ispitati učinak akutnoga izlaganja smjesi olova i kadmija na parametre oksidacijskoga stresa i status bioelemenata u mozgu štakora. Životinje su bile podijeljene u pet skupina: Cd15 (15 mg/kg), Cd30 (30 mg/kg), Pb150 (150 mg/kg), Pb150+Cd15 i kontrolna, netretirana skupina. Smjesa olova i kadmija dovela je do značajno viših vrijednosti malondialdehida (MDA) i drugih produkata u mozgu štakora u usporedbi s kontrolnom skupinom i obama pojedinačnim metalima. To je iznimno važno, imajući u vidu da je prisutnost MDA-a u mozgu povezana s mnogim neurodegenerativnim poremećajima. Nije dobivena statistički značajna razlika između ispitivanih skupina u razinama cinka (Zn), bakra (Cu), mangana (Mn) I željeza (Fe). Rezultati ovoga istraživanja važan su doprinos budućim istraživanjima smjesa i upućuju na to da istraživanja toksičnosti u kojima se procjenjuje neurotoksičnost pojedinačnih kemikalija ne pružaju stvarni uvid u neurotoksičnost njihovih smjesa kojima smo izloženi u stvarnom životu. Dobiveni rezultati otvaraju daljnja pitanja o interakciji i mehanizmima toksičnosti smjese tih metala

Procena unosa fluorida putem vode za piće i paste za zube kod dece uzrasta od tri godine - preliminarni rezultati u Beogradu, Republika Srbija

Background/Aim. Fluoride has beneficial effect on dental caries prevention and enables high hardness of enamel. However, fluoride intake above optimal levels can have adverse effects on teeth and bones, especially in young children during the period of intense growth and teeth development. The aim of this study was to assess fluoride intake from water and toothpaste among 3-year-old children in Belgrade, Serbia, in the municipalities of Vračar and Novi Beograd. Methods. A questionnaire for the parents (n = 40) was used to provide information on the water consumption (tap and/or bottled water) and the brand of toothpaste used by children as well as the frequency of tooth brushing and the amount of toothpaste during brushing. Fluoride concentrations in water and toothpaste samples were determined electrochemically by using fluoride-selective electrode. Fluoride intake was estimated through a mathematical model commonly used by the U.S. Environmental Protection Agency. Results. The obtained results indicate no significant difference in daily fluoride intake through drinking water and toothpaste in 3-year-old children in Vračar (n = 19) compared to Novi Beograd (n = 21) (p > 0.05). However, all estimated fluoride levels (0.089-0.625 mg/day) are significantly lower than the optimal daily intake level for caries protection (0.7 mg/day for children up to 4 years, FNB-USA National Institute of Medicine) and two to six times lower than tolerable upper fluoride level for the children of same age (1.3 mg/day, FNB-USA National Institute of Medicine). Furthermore, calculated daily fluoride intake per kilogram body weight confirm very low fluoride intake by water and toothpaste in children of investigated municipalities in Belgrade, being significantly below the recommended an adequate intake (0.05 mg/kg/day, EFSA). Conclusion. This preliminary study has shown that daily fluoride intake in 3-year-olds is lower than tolerable upper fluoride level, even not sufficient for the prevention of dental caries.Uvod/Cilj. Fluoridi imaju pozitivan efekat na prevenciju karijesa zuba kao i povećanje čvrstine zubnog emajla. Međutim, njihov unos iznad optimalnog može imati štetne efekte na zube i kosti, posebno kod male dece tokom intenzivnog rasta i razvoja zuba. Cilj ovog rada bio je da se proceni unos fluorida putem vode za piće i paste za zube kod dece uzrasta od tri godine u Beogradu, na teritoriji dve opštine, Vračar i Novi Beograd. Metode. Anketnim upitnicima (n = 40) za roditelje dobijeni su podaci o vrsti vode koju deca piju (česmenska i/ili flaširana), proizvodima koje koriste za čišćenje i negu zuba, učestalosti pranja zuba kao i količini paste za zube koja se koristi po jednom pranju. Sadržaj fluorida u vodi za piće i pastama za zube određen je elektrohemijski sa fluoridnom jonselektivnom elektrodom. Unos fluorida procenjen je korišćenjem matematičkog modela datog od strane Američke agencije za zaštitu životne sredine. Rezultati. Dobijeni rezultati pokazuju da nema statistički značajne razlike u dnevnom unosu fluorida putem vode za piće i paste za zube između dece uzrasta od tri godine na Vračaru i Novom Beogradu (p > 0,05). Međutim, sve procenjene vrednosti unosa fluorida (0,089-0,625 mg/dan) značajno su niže od optimalnog dnevnog unosa za prevenciju karijesa (0,7 mg/dan za decu uzrasta do 4 godine, FNB-Nacionalnog instituta medicine, SAD) i dva do šest puta niže od tolerišućeg gornjeg nivoa unosa fluorida za decu istog uzrasta (1,3 mg/dan, FNBNacionalnog instituta medicine, SAD). Štaviše, izračunati dnevni unosi fluorida izraženi po kilogramu telesne mase potvrđuju veoma nizak unos fluorida kod dece ispitivanih opština u Beogradu, čak značajno niže od preporučenog optimalnog unosa (0,05 mg/kg/dan, EFSA). Zaključak. Ovim preliminarnim istraživanjem pokazano je da je dnevni unos fluorida kod trogodišnjaka u Beogradu značajno niži od tolerišućeg gornjeg nivoa unosa fluorida, čak niži od optimalnog unosa za prevenciju karijesa

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

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

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

Vremenski zavisna pojava lažno pozitivnih rezultata test traka na amfetamin nakon primene OTC preparata sa pseudoefedrinom

Amphetamine is a potent stimulant of central nervous system and one of the most often abused substances. Immunochromatographic test panels for preliminary urineanalysis are used to track its abuse. Immunochromatographic analysis is quick, simple and based on the principle of competitive binding. Its main disadvantage is cross reactivity - tendency of antibodies to bind structurally similar compounds, including pseudoephedrine. This study aimed to examine the effect of pseudoephedrine-based OTC preparations on the occurrence of false-positive results of preliminary amphetamine urineanalysis and determine the time period during which these results may occur. The study included two subjects. TylolHot® was used for two days, while, on the first day, single dose was administered. On the second day, preparation was administered according to the dosing regimen recommended by the manufacturer (three times a day). Urine samples were collected for three consecutive days. Analysis was performed by immunochromatographic test strips "DIAQUICK" Multi-Drug Test Panels. TylolHot® use led to false positive results for amphetamine 3.75 h after single administration, and lasted for 27 h with repeated administration. After the application of the tested preparation, false positive results on amphetamine were obtained in different time periods, which depended on the intra- and interindividual variations, as well as the dosing regimen. Although test strips are used for quick preliminary analysis, it is necessary to check the results via confirmation methods (gas or liquid chromatography coupled to mass spectrometry). The obtained results are significant during routine testing of patients, employees, students, athletes and members of the army.Amfetamin je snažan stimulans centralnog nervnog sistema i jedna od supstanci koja se najčešće zloupotrebljavaju. Za praćenje njegove zloupotrebe koriste se imunohromatografske test trake za preliminarnu analizu urina. Imunohromatografska analiza je brza i jednostavna metoda koja se zasniva na principu kompetitivnog vezivanja. Njen glavni nedostatak je unakrsna reaktivnost – sklonost antitela da vezuju strukturno slična jedinjenja, između ostalih i pseudoefedrin. Cilj ovog rada bio je ispitati uticaj primene OTC preparata na bazi pseudoefedrina na pojavu lažno pozitivnih rezultata preliminarne analize amfetamina u urinu i utvrditi vremenski period pojave ovih rezultata od trenutka primene preparata. Ispitivanje je uključivalo dva ispitanika. Tokom dva dana korišćen je preparat TylolHot® , pri čemu je prvi dan primenjena jedna doza, a drugi dan se preparat primenjivao prema režimu doziranja preporučenom od strane proizvođača (tri puta dnevno). Uzorci urina sakupljani su tokom tri uzastopna dana, a analiza je izvršena imunohromatografskim test trakama "DIAQUICK" Multi-Drug Test Panels. Dobijeni su lažno pozitivni rezultati na amfetamin već u periodu od 3,75 h nakon jednokratne primene TylolHot ® preparata, i trajali su tokom 27 h pri višekratnoj primeni. Utvrđeno je da se nakon primene TylolHot® preparata u različitim vremenskim periodima mogu dobiti lažno pozitivni rezultati, što zavisi od intra- i interindividualnih varijacija i režima doziranja. Iako se test trake koriste za brzu preliminarnu analizu, neophodno je potvrditi rezultate konfirmativnim metodama (gasna ili tečna hromatografija spregnuta sa masenom spektrometrijom). Dobijeni rezultati su značajni prilikom rutinskih testiranja pacijenata, zaposlenih, učenika, sportista i pripadnika vojske.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

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