Ekspozom i endokrino zdravlje ljudi

The term exposome describes every exposure from the environment to which an individual is subjected from conception to death. Rapidly increasing evidence has documented that exposure to endocrine-disrupting chemicals (EDCs), an important part of the exposome, substantially contributes to disease and dysfunction across the life span. To date, however, there have been few studies investigating simultaneous exposure to multiple EDCs in mixtures, although this scenario entails real-life exposures. This paper will present the “Decoding the role of exposome in endocrine health” – DecodExpo project funded by the Science Fund of the Republic of Serbia. The project aims to provide scientific evidence for the role of toxic metal(oid)s mixtures (As, Pb, Hg, Cd, Cr, and Ni) in the development of various endocrine disorders. The project consists of human biomonitoring (HBM), exposome-wide association, and animal studies. Namely, data on blood levels of toxic metal(oid)s in different cohort groups within the Serbian general population were collected (435 participants) and, by performing mathematical modelling, dose-response relationships were determined between certain toxic metals and hormone blood levels. The study protocol for the animal study was chosen based on the results of the HBM study, to address the main challenges in studies on EDCs (non-monotonic dose-response, long-term exposure to low doses, exposure to mixtures of EDCs, sex differences). Furthermore, various endpoints are tested to ensure better insight into molecular, biochemical, and structural changes in the endocrine system. The project revealed an important role of exposure to the investigated toxic metal(oid)s mixture in human endocrine health.Izraz ekspozom opisuje sve uticaje iz životne sredine kojima je pojedinac izložen od začeć a do smrti. Brojni naučni dokazi ukazuju da izloženost hemikalijama koje ometaju rad endokrinog sistema, endokrinim ometačima, važnim komponentama ekspozoma, značajno doprinosi nastanku bolesti i različitim poremećajima tokom životnog veka. Međutim, malo je studija koje istražuju istovremenu izloženost endokrinim ometačima koji su u prirodi prisutni u smešama, iako baš ovaj scenario odslikava izloženost u stvarnom životu. Ovo predavanje daje kratak prikaz projekta „Dekodiranje uloge ekspozoma u endokrinom zdravlju” – DecodExpo koji finansira Fond za nauku Republike Srbije u okviru PROMIS programa. Projekat ima za cilj da pruži naučne dokaze o ulozi smeše toksičnih metal(oid)a (As, Pb, Hg, Cd, Cr, i Ni) u razvoju različitih endokrinih poremeć aja i bolesti. Ovo istraživanje obuhvatilo je studije humanog biomonitoringa (HBM), ispitivanje veze ekspozoma i zdravlje, te studije na životinjama. Naime, prikupljeni su podaci o nivoima toksičnih metala u krvi kod ispitanika iz opšte populacije Srbije (435 učesnika) i matematičkim modeliranjem utvrđeni su odnosi doza-odgovor između nivoa pojedinih metal(oida)a i nivoa hormona u krvi. Protokol studije na životinjama izabran je na osnovu rezultata HBM studije, kako bi se odgovorilo na glavne izazove sa kojima se naučnici suočavaju pri sprovođenju studija o endokrinim ometačima (nemonotoni odnos doza-odgovor, dugotrajno izlaganje niskim dozama, izloženost smešama, razlike među polovima). Pored toga, ispitani su različiti parametri kako bi se obezbedio bolji uvid u molekularne, biohemijske i strukturne promene u endokrinom sistemu. Projekat otkriva važnu ulogu koju izloženost ovoj toksičnoj smeši metal(oid)a ima u ljudskom endokrinom zdravlju.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Lekovi endokrini ometači u životnoj sredini ‐ pretnja po zdravlje ljudi i vodene vrste?

The potential negative effects of pharmaceuticals, especially those with endocrine disrupting (ED) properties, present in the environment has been attracting considerable attention in recent years. These pharmaceuticals enter the environment via anthropogenic activities; however, limited data are available on their fate, behaviour, and pathways. Literature data point to adverse ecological effects of these compounds for example, the feminization of aquatic species due to estrogenic compounds present in different aquatic compartments (1). However, there is no consensus on what risk, if any, these compounds pose to human health. Namely, a lot has been discussed about potential estrogenicity of drinking water and the possible contribution of this phenomenon in the observed rise in human reproductive problems. Although estimations pointed to a negligible risk from synthetic estrogens in drinking water (2), these chemicals have been detected in drinking water at nanogram per litre to microgram per litre concentrations which are the concentrations that potentially can produce adverse effects in humans leading to various endocrine disorders (3). Moreover, it must be considered that humans and other organisms are often exposed to various chemicals mixtures of unknown composition and effects, hence, making the assessment based on a single substance sometimes inadequate. It can be concluded that the presence of pharmaceuticals with ED properties is a rather complex phenomena difficult to predict both due to various pathways they can have in the environment and multiplicity of the effects they can have on human health. However, the evidence collected worldwide are confirming their presence in the environment; hence, future studies need to be pointed towards a better understanding of their fate in the environment, effects of low doses of EDCs exposure and endocrine disorders they can trigger in both animals and humans. Special attention should be also given to the identification of the strengths and limitations of current analytical methods used for their detection and further development of treatment technologies for their removal from water. It is evident that in the future, pharmaceutical research should be pointed not only towards the pharmaceuticals’ development, but also towards the development of risk-based models that can predict the potential sources, fate, behaviour and effects of pharmaceuticals once they enter the environment.Potencijalni štetni efekti lekova, posebno onih sa svojstvima endokrinih ometača, prisutnih u životnoj sredini, poslednjih godina privlače sve više pažnje. Ovi lekovi ulaze u životnu sredinu kao posledica antropogenih aktivnosti, ali njihova sudbina i ponašanje, nakon što se nađu u životnoj sredini, i dalje predstavlja nepoznanicu. Literaturni podaci ukazuju na štetne ekološke efekte ovih jedinjenja, na primer, utvrđeno je da estrogena jedinjenja prisutna u različitim vodenim delovima ekosistema izaziva feminizaciju vodenih vrsta (1). Međutim, ne postoji konsenzus o tome kakav rizik ove supstance predstavljaju za ljudsko zdravlje. Naime, mnogo se govori o potencijalnoj estrogenosti vode za piće i mogućoj ulozi ove pojave u uočenom porastu reproduktivnih problema kod ljudi. Iako su procene ukazivale na zanemarljiv rizik od prisustva sintetičkih estrogena u vodi za piće (2), ove hemikalije su ipak izmerene u vodi za pić e u koncentracijama od nekoliko nanograma po litru do nekoliko mikrograma po litru, što su koncentracije koje potencijalno mogu izazvati štetne efekte na ljude i dovesti do različitih endokrinih poremećaja (3). Takođe, mora se uzeti u obzir da su ljudi i drugi organizmi često izloženi različitim smešama hemikalija nepoznatog sastava i efekata, pa je procena zasnovana na jednoj supstanci ponekad neadekvatna. Može se zaključiti da je prisustvo lekova sa svojstvima endokrinih ometača u životnoj sredini prilično kompleksna pojava čije je efekte i posledice teško predvideti kako zbog različitih puteva koje mogu imati u životnoj sredini, tako i zbog višestrukih efekata koje mogu imati na zdravlje ljudi. Međutim, dokazi prikupljeni širom sveta potvrđuju njihovo prisustvo u okruženju, pa je buduće studije potrebno usmeriti ka boljem razumevanju njihove sudbine u životnoj sredini, te efekata izloženosti niskim dozama i potencijalnih endokrinih poremeć aja koje mogu izazvati kako kod životinja, tako i kod ljudi. Posebnu pažnju treba posvetiti i identifikaciji ograničenja postojeć ih analitičkih metoda koje se koriste za utvrđivanje prisustva ovih supstanci u vodi, kao i daljem razvoju tretmana za njihovo uklanjanje iz vode. Očigledno je da bi u budućnosti farmaceutska istraživanja trebalo da budu usmerena ne samo na razvoj farmaceutskih proizvoda, već i na razvoj modela koji mogu predvideti potencijalne izvore, sudbinu, ponašanje i efekte lekova kada se nađu u životnoj sredini.Drugi naučni simpozijum Saveza farmaceutskih udruženja Srbije sa međunarodnim učešćem, 28. 10. 2021. Beogra

Dekodiranje uloge koktela toksičnih metala u endokrinom zdravlju: studija na opštoj populaciji Republike Srbije

The human population is exposed to a vast number of chemicals that are known or suspected endocrine disruptors. Although thresholds are mainly determined concerning a single compound, in real life we are indeed exposed to multiple chemicals in mixtures, i.e. toxic cocktails. The focus of this research was to determine the effects that toxic cocktails of metal(oid)s have on human endocrine health The study involved 435 participants from the general population of Serbia (218 women and 217 men) from five different cohorts (prostate and testes cancer patients, breast cancer and benign breast dysplasia, pancreatic cancer, thyroid, and metabolic disorders patients, and healthy volunteers). Blood cadmium (Cd), lead (Pb), arsenic (As), mercury (Hg), nickel (Ni), and chromium (Cr) levels and serum levels of various hormones were measured in all collected samples. The statistical analyses were performed using the R language while dose-response analysis was performed in PROAST software. Levels estimated to produce the 10% extra risk of testosterone serum levels disturbances were lower than median Hg levels measured in the general population. The obtained results also provided support for the positive association between Pb exposure and higher insulin levels. Similarly, the calculated levels that can produce 10% of extra risk for the disturbances in free thyroxine levels were lower than the medium determined As levels. These findings provide strong support for the endocrine-disrupting effects of the investigated toxic cocktail of metal(oid)s present in our environment and suggest that threshold values for such effects might be lower than those currently implemented.Opšta populacija je svakodnevno izložena velikom broju hemikalija za koje se zna ili se sumnja da su endokrini ometači. Iako se referentne doze uglavnom određuju za pojedinačne hemikalije, u stvarnom životu smo zapravo izloženi hemikalijama u smešama, odnosno toksičnim koktelima. Fokus istraživanja bio je na utvrđivanju uticaja koje toksični koktel metala(oida) imaju na endokrino zdravlje ljudi. Istraživanje je obuhvatilo 435 učesnika iz opšte populacije Republike Srbije (218 žena i 217 muškaraca) iz pet različitih kohorti (pacijenti sa: karcinomima prostate ili testisa, karcinomom dojke ili benignom displazijom dojke, karcinomom pankreasa, poremećajima štitaste žlezde i metaboličkim poremeć ajima, te zdravi dobrovoljci). U prikupljenim uzorcima mereni su nivoi kadmijuma (Cd), olova (Pb), arsena (As), žive (Hg), nikla (Ni) i hroma (Cr) u krvi, kao i nivoi različitih hormona u serumu. Statističke analize su obavljene korišćenjem R statističkog paketa, dok je doza-odgovor analiza vršena u PROAST softveru (RIVM, Holandija). Nivoi za koje se procenjuje da dovode do dodatnog rizika od 10% za nastanak poremeć aja nivoa testosterona u serumu bili su niži od medijane izmerenih nivoa Hg kod ispitanika. Dobijeni rezultati su takođe pružili podršku postojanju veze između nivoa izloženosti Pb i viših nivoa insulina. Slično, izračunati nivoi As u krvi koji mogu da izazovu 10% dodatnog rizika za poremeć aje nivoa slobodnog tiroksina bili su niži od izračunate medijane za nivoe ovog metaloida u krvi. Ovi nalazi pružaju dokaze o endokrinim efektima ispitivanog koktela toksičnih metal(oid)a i sugerišu da bi granične vrednosti za takve efekte mogle biti niže od onih koje se trenutno primenjuju.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

The Validity of Benchmark Dose Limit Analysis for Estimating Permissible Accumulation of Cadmium

Cadmium (Cd) is a toxic metal pollutant that accumulates, especially in the proximal tubular epithelial cells of kidneys, where it causes tubular cell injury, cell death and a reduction in glomerular filtration rate (GFR). Diet is the main Cd exposure source in non-occupationally exposed and non-smoking populations. The present study aimed to evaluate the reliability of a tolerable Cd intake of 0.83 μg/kg body weight/day, and its corresponding toxicity threshold level of 5.24 μg/g creatinine. The PROAST software was used to calculate the lower 95% confidence bound of the benchmark dose (BMDL) values of Cd excretion (ECd) associated with injury to kidney tubular cells, a defective tubular reabsorption of filtered proteins, and a reduction in the estimated GFR (eGFR). Data were from 289 males and 445 females, mean age of 48.1 years of which 42.8% were smokers, while 31.7% had hypertension, and 9% had chronic kidney disease (CKD). The BMDL value of ECd associated with kidney tubular cell injury was 0.67 ng/L of filtrate in both men and women. Therefore, an environmental Cd exposure producing ECd of 0.67 ng/L filtrate could be considered as Cd accumulation levels below which renal effects are likely to be negligible. A reduction in eGFR and CKD may follow when ECd rises from 0.67 to 1 ng/L of filtrate. These adverse health effects occur at the body burdens lower than those associated with ECd of 5.24 µg/g creatinine, thereby arguing that current health-guiding values do not provide a sufficient health protection

The NOAEL Equivalent of Environmental Cadmium Exposure Associated with GFR Reduction and Chronic Kidney Disease

Cadmium (Cd) is a highly toxic metal pollutant present in virtually all food types. Health guidance values were established to safeguard against excessive dietary Cd exposure. The derivation of such health guidance figures has been shifted from the no-observed-adverse-effect level (NOAEL) to the lower 95% confidence bound of the benchmark dose (BMD), termed BMDL. Here, we used the PROAST software to calculate the BMDL figures for Cd excretion (ECd) associated with a reduction in the estimated glomerular filtration rate (eGFR), and an increased prevalence of chronic kidney disease (CKD), defined as eGFR ≤ 60 mL/min/1.73 m2. Data were from 1189 Thai subjects (493 males and 696 females) mean age of 43.2 years. The overall percentages of smokers, hypertension and CKD were 33.6%, 29.4% and 6.2%, respectively. The overall mean ECd normalized to the excretion of creatinine (Ecr) as ECd/Ecr was 0.64 µg/g creatinine. ECd/Ecr, age and body mass index (BMI) were independently associated with increased prevalence odds ratios (POR) for CKD. BMI figures ≥24 kg/m2 were associated with an increase in POR for CKD by 2.81-fold (p = 0.028). ECd/Ecr values of 0.38–2.49 µg/g creatinine were associated with an increase in POR for CKD risk by 6.2-fold (p = 0.001). The NOAEL equivalent figures of ECd/Ecr based on eGFR reduction in males, females and all subjects were 0.839, 0.849 and 0.828 µg/g creatinine, respectively. The BMDL/BMDU values of ECd/Ecr associated with a 10% increase in CKD prevalence were 2.77/5.06 µg/g creatinine. These data indicate that Cd-induced eGFR reduction occurs at relatively low body burdens and that the population health risk associated with ECd/Ecr of 2.77–5.06 µg/g creatinine was not negligible

Exploring the relationship between blood toxic metal(oid)s and serum insulin levels through benchmark modelling of human data: Possible role of arsenic as a metabolic disruptor

The major goal of this study was to estimate the correlations and dose-response pattern between the measured blood toxic metals (cadmium (Cd), mercury (Hg), chromium (Cr), nickel (Ni))/metalloid (arsenic (As)) and serum insulin level by conducting Benchmark dose (BMD) analysis of human data. The study involved 435 non-occupationally exposed individuals (217 men and 218 women). The samples were collected at health care institutions in Belgrade, Serbia, from January 2019 to May 2021. Blood sample preparation was conducted by microwave digestion. Cd was measured by graphite furnace atomic absorption spectrophotometry (GF-AAS), while inductively coupled plasma-mass spectrometry (ICP-MS) was used to measure Hg, Ni, Cr and As. BMD analysis of insulin levels represented as quantal data was done using the PROAST software version 70.1 (model averaging methodology, BMD response: 10%). In the male population, there was no correlation between toxic metal/metalloid concentrations and insulin level. However, in the female population/whole population, a high positive correlation for As and Hg, and a strong negative correlation for Ni and measured serum insulin level was established. BMD modelling revealed quantitative associations between blood toxic metal/metalloid concentrations and serum insulin levels. All the estimated BMD intervals were wide except the one for As, reflecting a high degree of confidence in the estimations and possible role of As as a metabolic disruptor. These results indicate that, in the case of As blood concentrations, even values higher than BMD (BMDL): 3.27 (1.26) (male population), 2.79 (0.771) (female population), or 1.18 (2.96) μg/L (whole population) might contribute to a 10% higher risk of insulin level alterations, meaning 10% higher risk of blood insulin increasing from within reference range to above reference range. The obtained results contribute to the current body of knowledge on the use of BMD modelling for analysing human data

Toksičnost organskog i neogranskog nikla u ćelijskim kulturama pankreasa: poređenje sa kadmijumom

Nickel compounds are Group 1 carcinogens and possibly cancer-causing in the pancreas. We examined the toxicity of nickel in both 2-D and 3-D pancreatic cell cultures, to determine the LD50 for organic and inorganic nickel in normal and cancerous cells. Assays with cadmium chloride were performed to be a comparison to potential nickel-induced toxicity. Cells were exposed to twelve concentrations of NiCl2 or Ni-(Ac)2 for 48h (2-D), or six concentrations for 48 hours (3-D). There was a significant (P=0.0016) difference between HPNE and AsPC-1 LD50values after cadmium exposure, at 69.9 μM and 29.2 μM, respectively. Neither form of nickel exhibited toxicity in 2-D or 3-D cultures, but after 48h, changes in spheroid morphology were observed. The inability of Ni to reduce viable cell numbers suggests a toxic mechanism that differs from cadmium, also a Group 1 carcinogen. The cell microenvironment was not a factor in nickel toxicity with no changes in viable cells in either 2-D or 3-D cultures. These studies only examined cytotoxicity, and not genotoxicity, a potential mechanism of nickel carcinogenicity. Alterations in DNA function or the expression of apoptotic proteins/processes would take longer to manifest. Current work focuses on cellular changes following extended nickel exposure.Uloga nikla, toksičnog metala, u nastanku karcinoma pankreasa još uvek nije u potpunosti ispitana. Cilj rada je da ispita toksičnost nikla (Ni) u 2-D i 3-D kulturi ćelija pankreasa, kako bi se utvrdila LD50 vrednost za organski i neorganski nikl u normalnim i tumorskim ćelijama. Ispitivanja su izvršena i sa kadmijumom (Cd), metalom čija je uloga u nastanku karcinoma pankreasa potvrđena u prethodnim istraživanjima, a u svrhu poređenja sa potencijalnom toksičnošću izazvanom Ni. Ćelije su bile tretirane sa 12 različitih koncentracija NiCl2 ili Ni-(Ac)2 tokom 48h (2-D), odnosno sa šest različitih koncentracija tokom 48 sati (3-D). Nijedan oblik Ni nije ispoljio toksičnost u 2-D ili 3-D kulturama, ali nakon 48h primećene su promene u sferoidnoj morfologiji. Nemogućnost Ni da smanji broj vijabilnih ćelija sugeriše mehanizam karcinogeneze različit od mehanizma koji ispoljava Cd. Ipak, da bi se uočile izmene u funkciji DNK ili izražavanju apoptotičkih proteina/procesa potrebno je duže vremena, pa su dalja istraživanja upravo fokusirana na ćelijske promene nakon produžene izloženosti nikla

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

Potential interaction of cadmium chloride with pancreatic mitochondria: Implications for pancreatic cancer

Pancreatic cancer (PC) is insidious with a high mortality rate due to the lack of symptomology prior to diagnosis. Mitochondrial involvement in PC development is becoming accepted, and exposure to cadmium (Cd) is suspected of being a risk factor for the development of PC; however, the mechanisms involved remain unclear. In this study, we examined the role of Cd as a mitochondrial toxicant and whether alterations in mitochondrial function may be an underlying cause for the development of PC. In this study, cadmium chloride (CdCl2)-mediated toxicity in hTERT-HPNE and AsPC-1 pancreatic cell lines was determined by MTT assay. We also investigated the release of LDH and the generation of free radicals. Mitochondrial toxicity assays were performed in media containing glucose (25 mM) or galactose (10 mM) and following exposure to CdCl2 (0-100 M) followed by MTT assay. For the confirmation of mitochondrial toxicity, we measured the release of ATP following exposure to CdCl2. Initial experiments confirmed that exposure to CdCl2 did not reduce the viability of either cell line until a concentration of >10 M was used. Non-linear analysis of the response curves revealed lethal concentration 50% (LC50) values for CdCl2 in the HPNE cells of 77 M compared to 42 M in the AsPC-1 cells (P lt 0.01). The CdCl2-mediated mitochondrial toxic effects were greater in the HPNE cells, suggesting a heightened sensitivity to the effects of CdCl2, not due to elevated oxidative stress. Increased mitochondrial toxic sensitivity was indicated by a 73.4% reduction in IC50 values in the HPNE cells cultured in galactose compared to culture in glucose media, whereas the AsPC-1 cells exhibited a 58.8% reduction in IC50 values. In addition, the higher concentration of CdCl2 elicited a significant cell-dependent effect on ATP release in both cell lines, suggestive of CdCl2 being a mitochondrial toxicant. Cell survival was unaffected following exposure to low concentrations of CdCl2; however, exposure did alter mitochondrial function (control cells > tumor cells). Therefore, the findings of this study indicate that the mitochondria may be a site of action for cadmium in promoting tumor development