Toksikološki aspekt imunomodulatora: prijatelj ili neprijatelj u povećanju efikasnosti imunoterapije karcinoma

Malignant diseases world incidence is constantly increasing and it is assumed that combining immunomodulators with immunotherapy would improve cancer therapy effectiveness. We present a research-development project of international cooperation with the People's Republic of China entitled "Increasing the effectiveness of cancer immunotherapy with a combination of CAR-T cells or PD-1/PD-L1 inhibitors using immunomodulators", led by Faculty of Pharmacy, University of Belgrade. Sulforaphane (SFN), isothiocyanate from cabbage vegetables, and a type of inactivated bacterium Pseudomonas aeruginosa (PA-MSHA) have been recognized as immunomodulators with high immune system stimulating potential, i.e. antitumor effect. However, there are few data on their individual safe use, and no data on the potential side/harmful effects of their combination, especially in patients with malignant diseases and significant immune system impairment. The Chinese team is investigating the effectiveness of improving immunotherapy with combination of immunomodulators, with their first published results proving SFN positive effect when administered with CAR-T cells (1), while Serbian team aims to examine toxicological profiles of single and/or combined SFN and PA MSHA, in silico, in vitro and in vivo on two experimental models - zebrafish and rat. The project will specifically examine adverse effects potential of single and/or combined use of the tested immunomodulators or lack of their efficacy in cancer patients, especially colon cancer. The first in silico results of the Serbian team indicated benefit/risk of SFN in patients with colon cancer depending on individual genes expression and identified gene set which change may indicate a positive or negative effect of immunomodulators (2).Incidenca malignih oboljenja u svetu je u stalnom porastu i pretpostavlja se da bi kombinacija imunomodulatora sa imunoterapijom poboljšala efikasnost terapije kod pacijenata obolelih od raka. Ovaj rad prezentuje istraživačko-razvojni projekat međunarodne saradnje sa NR Kinom pod nazivom „Povećanje efikasnosti imunoterapije karcinoma kombinacijom CAR-T ćelija ili PD-1/PD-L1 inhibitora pomoću imunomodulatora” čiji je nosilac Farmaceutski fakultet Univerziteta u Beogradu. Sulforafan (SFN), izotiocijanat iz kupusastog povrća, i vrsta inaktivisane bakterije Pseudomonas aeruginosa (PA-MSHA) prepoznati su kao imunomodulatori sa velikim stimulativnim potencijalom imunskog sistema, odnosno antitumorskim efektom. Međutim, malo je podataka o njihovoj pojedinačnoj bezbednoj primeni, dok nema podataka o potencijalnim neželjenim i štetnim efektima njihove kombinovane primene, posebno kod pacijenata obolelih od malignih bolesti sa značajno narušenim imunskim sistemom. Kineski tim ispituje efikasnost poboljašanja imunoterapije kombinacijom sa imunomodulatorima, pri čemu je njihovim prvim publikovanim rezultatima dokazano pozitivno dejstvo SFN kada se primenjuje sa CAR-T ćelijama (1), dok je zadatak srpskog tima da ispita toksikolški profile pojedinačne ili kombinovane primene SFN i PA-MSHA in silico, in vitro i in vivo na dva eksperimentalna modela – zebrica i pacova. Projekat će posebno ispitati potencijal za pojavu štetnih efekata pojedinačne i/ili kombinovane primene ispitivanih imunomodulatora ili pak izostanak efikasnosti terapije kod pacijenata sa rakom, posebno rakom debelog creva. Prvi in silico rezultati srpskog istraživačkog tima ukazuju da korist ili rizik primene SFN kod pacijenata sa rakom debelog creva zavisi od ekspresije pojedinih gena kod pacijenta i identifikuje setove gena čija promena može ukazati na pozitivan ili čak negativan efekat primene imunomodulatora (2).VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

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

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

Zašto je potrebna toksikološka procena rizika za kozmetički proizvod?

Bearing in mind that cosmetic products are applied throughout the lifetime and that they can contain potentially harmful and toxic substances, valid EU regulation in this area has set strict requirements regarding their safety. Manufacturers are required to provide a cosmetic product safety report containing a toxicological assessment made by experts. Toxicological assessment is based on existing in vivo toxicological data, and in particular in silico and in vitro, since EU regulations in the field of cosmetic products prohibit new animal studies. Although sporadic cases of serious poisoning after the application of cosmetic products have been recorded, adverse effects after normal and reasonably foreseeable use of cosmetic products are usually mild (allergic reactions and skin irritation) and completely reversible. EU regulations require mandatory reporting of serious adverse effects (functional incapacity, hospitalization, disability, etc.) in order to detect ingredients of cosmetic products that potentially endanger human health and prevent similar effects from recurred. The paper describes the procedure of toxicological risk assessment for cosmetic ingredients because the assessment of the safety of the cosmetic product is based on the assessment of the safety of its ingredients.Imajući u vidu da se kozmetički proizvodi primenjuju tokom celog života a da mogu sadržati potencijalno štetne i toksične supstance, važeća regulativa EU u ovoj oblasti (Uredba (EC) 1223/2009) postavila je stroge zahteve u pogledu njihove bezbednosti. Proizvođači su u obavezi da dostave izveštaj o bezbednosti koji sadrži toksikološku procenu koju rade osobe kvalifikovane za ovu vrstu poslova. Toksikološka procena se vrši na osnovu postojećih podataka in vivo toksikoloških ispitivanja, i posebno in silico i in vitro budući da EU regulativa u oblasti kozmetičkih proizvoda zabranjuje nova ispitivanja na životinjama. Iako se sporadično javljaju i ozbiljni slučajevi trovanja nakon primene kozmetičkih proizvoda, neželjeni efekti pri uobičajenoj i predvidivoj upotrebi kozmetičkog proizvoda su obično blagi (alergijske reakcije i iritacija kože) i potpuno reverzibilni. Regulativa EU zahteva obavezno prijavljivanje ozbiljnih neželjenih efekata (privremena sprečenost za rad, hospitalizacija, invaliditet itd.) u cilju otkrivanja sastojaka kozmetičkih proizvoda koji potencijalno ugrožavaju zdravlje ljudi i preduzimanja određenih mera kako bi se sprečilo da se slični efekti ponovo jave. U radu je opisana procedura toksikološke procene rizika za kozmetičke sastojke jer se procena bezbednosti kozmetičkog proizvoda zasniva na proceni bezbednosti njegovih sastojaka

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

Effects of oral and intraperitoneal magnesium treatment against cadmiuminduced oxidative stress in plasma of rats

Cadmium (Cd) has been recognised as one of the most important environmental and industrial pollutants, and up-to-date investigations have shown that one of the mechanisms of its toxicity is associated with the induction of oxidative stress. The aim of this study was to determine the connection between acute oral and intraperitoneal exposure to Cd and parameters indicative of oxidative stress in the plasma of rats, as well as to examine the potential protective effect of magnesium (Mg) in conditions of acute oral and intraperitoneal Cd poisoning. The experiment was performed on male albino Wistar rats (n=40) randomly divided into control group, Cd-or group that received 30 mg kg(-1) b.w. Cd by oral gavage, Cd+Mg-or group that orally received 50 mg kg(-1) b.w. Mg one hour before oral Cd, Cd-ip group that received 1.5 mg kg(-1) b.w. Cd intraperitoneally, and Cd+Mg-ip group that intraperitoneally received 3 mg kg(-1) b.w. Mg 10 min before intraperitoneal Cd. The animals were sacrificed 24 h after treatment and the following parameters were measured: superoxide-dismutase activity, superoxide anion, total oxidative status, advanced oxidation protein products, and malondialdehyde. All parameters of oxidative stress in rat plasma were negatively affected by Cd treatment with more pronounced negative effects after intraperitoneal treatment, with the exception of superoxide dismutase (SOD) activity. Although both oral and intraperitoneal Mg pretreatment had protective effects, more pronounced beneficial effects were observed after oral administration, since it managed to completely prevent Cd-induced changes in the investigated parameters. The observed results support the use of Mg as potential protective agent against toxic effects caused by Cd

Effect of Magnesium Supplementation on the Distribution Patterns of Zinc, Copper, and Magnesium in Rabbits Exposed to Prolonged Cadmium Intoxication

The present study is designed to investigate whether magnesium (Mg) supplementation may prevent Cd-induced alterations in zinc (Zn), copper (Cu), and magnesium (Mg) status in rabbits. For this purpose, the concentrations of Zn, Cu, and Mg were estimated in blood, urine, and organs (brain, heart, lungs, liver, kidney, spleen, pancreas, skeletal muscle, and bone) of rabbits given Cd (10 mg/kg b.w.) and rabbits cotreated with Mg (40 mg/kg b.w.) orally, as aqueous solutions of Cd chloride and Mg acetate every day for 4 weeks. Samples were mineralized with conc. HNO3 and HClO4 (4:1) and metals concentrations were determined by atomic absorption spectrophotometry (AAS). Magnesium supplementation succeeded to overcome Cd-induced disbalance of investigated bioelements. Beneficial effects of Mg were observed on Zn levels in blood and urine, on Cu levels in urine, and on Mg levels in blood. Magnesium pretreatment also managed to counteract or reduce all Cd-induced changes in levels of Cu and Mg in organs, while it did not exert this effect on Zn levels. These findings suggest that enhanced dietary Mg intake during Cd exposure can have at least partly beneficial effect on Cd-induced alterations in homeostasis of zinc, copper, and magnesium

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

The role of toxic metals and metalloids in nrf2 signaling

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

Sadržaj fluorida u flaširanim mineralnim vodama Srbije

The beneficial effects of fluoride, especially in reducing dental caries, are well known (recommended value 1mg/L of drinking water). However, prolonged intake of fluoride levels exceeding 1,5 mg/L can cause detrimental effects, mostly on teeth and skeleton. Since water is one of the principal sources of fluoride intake, the objective of this study was to determine fluoride content in bottled mineral waters of Serbia. 20 samples of different bottled mineral waters produced in Serbia were analyzed. Fluoride levels were determined electrochemically, using fluoride-selective electrode, after mixing with TISAB buffer in ratio 1:1. In 3 of 20 samples of bottled mineral waters fluoride levels exceeded 1 mg/L - carbonated mineral water 'Vrnjci' (1,34 mg/L), 'Heba' (1,19 mg/L) and 'Knjaz Miloš' (1,15 mg/L). Lower fluoride content was detected in 'Voda Voda' (0,64 mg/L), 'Minaqua' (0,43 mg/L) and 'Mivela' (0,43 mg/L). Other analyzed mineral waters contained very low fluoride concentration, lower than 0,25 mg/l; 'Aqua Gala' (0,23 mg/L), 'Aqua Bella' (0,2 mg/L), 'Life' (0,19 mg/L), 'Aqua viva' (0,14 mg/L), 'Prolom voda' (0,12 mg/L), 'Vujić voda' (0,1 mg/L), 'Vrnjačko vrelo' (0,09 mg/L), 'Aqua Balkanika' (0,07 mg/L), 'Rosa'-carbonated (0,06 mg/L), 'Rosa'-non-carbonated (0,06 mg/L), 'Vrnjci'-non-carbonated (0,06 mg/L), 'Zlatibor'-carbonated (0,04 mg/L) and 'Zlatibor'-non-cabonated (0,04 mg/L). These results show that carbonated mineral water 'Vrnjci', 'Heba' and 'Knjaz Miloš' contain fluoride in concentrations similar to recommended value, so they can induce beneficial effects in reducing dental caries. Most of other analysed bottled waters contain very low fluoride concetrations, indicating that the consumers of those waters need additional fluoride intake via food or fluoride preparations.Pozitivni efekti fluorida, naročito u suzbijanju karijesa, dobro su poznati (preporučeni sadržaj u pijaćoj vodi iznosi 1 mg/L). Međutim, produženi unos fluorida, u koncentracijama višim od 1,5 mg/L vode može dovesti do neželjenih efekata, koji se ispoljavaju uglavnom na zubima i skeletu. Obzirom da je voda jedan od najznačajnijih izvora unošenja fluorida, cilj ovog rada je bio određivanje sadržaja fluorida u flaširanim mineralnim vodama Srbije. Analizirano je 20 uzoraka flaširanih mineralnih voda proizvedenih u Srbiji. Sadržaj fluorida je određen elektrohemijski, korišćenjem fluoridne jon-selektivne elektrode. Koncentracija fluorida je viša od 1 mg/L u 3 od 20 analiziranih flaširanih mineralnih voda - u gaziranoj mineralnoj vodi 'Vrnjci' (1,34 mg/L), i vodama 'Heba' (1,19 mg/L) i 'Knjaz Miloš' (1,15 mg/L). Niži sadržaj detektovan je u mineralnim vodama 'Voda voda' (0,64 mg/L), 'Minaqua' (0,43 mg/L) i 'Mivela' (0,43 mg/L), a u izuzetno niskim koncentracijama, nižim od 0,25 mg/L, fluoridi se nalaze u mineralnim vodama 'Aqua Gala' (0,23 mg/L), 'Aqua Bella' (0,2 mg/L), 'Life' (0,19 mg/L), 'Aqua viva' (0,14 mg/L), 'Prolom voda' (0,12 mg/L), 'Vujić voda' (0,1 mg/L), 'Vrnjačko vrelo' (0,09 mg/L), 'Aqua Balkanika' (0,07 mg/L), 'Rosa'-gazirana (0,06 mg/L), 'Rosa'-negazirana (0,06 mg/L), 'Vrnjci'-negazirana (0,06 mg/L), 'Zlatibor'-gazirana (0,04 mg/L) i 'Zlatibor'-negazirana (0,04 mg/L). Dobijeni rezultati ukazuju da gazirana mineralna voda 'Vrnjci', 'Heba' i 'Knjaz Miloš' sadrže fluoride u koncentracijama bliskim optimalnim vrednostima, te mogu ispoljiti pozitivan efekat u prevenciji karijesa. Većina ostalih ispitivanih flaširanih voda ima veoma nizak sadržaj fluorida, pa bi osobe koje konzumiraju ovakve vode trebalo da obezbede potreban unos fluorida putem hrane ili preparata fluorida