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

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

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

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

Utjecaj adulteranata u mokraći na nalaze komercijalnih testnih traka za otkrivanje zloporabe droga

Immunochromatographic strips for urine drug screening tests (UDSTs) are common and very suitable for drug abuse monitoring, but are also highly susceptible to adulterants kept in the household, which can significantly alter test results. The aim of this study was to see how some of these common adulterants affect UDST results in practice and whether they can be detected by sample validity tests with pH and URIT 11G test strips. To this end we added household chemicals (acids, alkalis, oxidizing agents, surfactants, and miscellaneous substances) to urine samples positive for amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), tetrahydrocannabinol, heroin, cocaine, or benzodiazepines (diazepam or alprazolam) and tested them with one-component immunochromatographic UDST strips. The UDST for cocaine resisted adulteration the most, while the cannabis test produced the most false negative results. The most potent adulterant that barely changed the physiological properties of urine specimens and therefore escaped adulteration detection was vinegar. Besides lemon juice, it produced the most false negative test results. In conclusion, some urine adulterants, such as vinegar, could pass urine specimen validity test and remain undetected by laboratory testing. Our findings raise concern about this issue of preventing urine tampering and call for better control at sampling, privacy concerns notwithstanding, and better sample validity tests.Preliminarna analiza prisutnosti psihoaktivnih tvari u mokraći pomoću imunokromatografskih testnih traka (UDST) našla je primjenu u mnogim područjima. Iako vrlo prikladne u kontroli zloporabe droga, testne su trake iznimno osjetljive na sredstva za patvorenje (adulterante) kao što su kućne kemikalije, što može značajno promijeniti rezultate testa. Cilj ovoga istraživanja bio je ispitati potencijal uobičajeno korištenih adulteranata kada je riječ o utjecaju na rezultate probira na prisutnost psihoaktivnih tvari u mokraći uporabom UDST-a. Ispitivane kemikalije (kiseline, lužine, oksidirajuća sredstva, površinski aktivne tvari i druge) dodane su uzorcima urina u kojima je, tekućinskom kromatografijom – masenom spektrometrijom (LC-MS) prethodno potvrđena prisutnost amfetamina, 3,4-metilendioksimetamfetaina (MDMA), tetrahidrokanabinola, heroina i kokaina benzodiazepina (diazepam ili alprazolam). U ispitivanju su korištene jednokomponentne imunokromatografske testne trake. Manipulacija uzorcima urina provjeravala se pomoću pH traka, kao i biokemijskih testnih traka za semikvantitativno određivanje koncentracija endogenih tvari i specifične težine uzoraka urina pomoću testnih traka URIT 11G. Rezultati ovoga ispitivanja pokazali su da je test za detekciju kokaina u urinu najmanje osjetljiv na utjecaj ispitivanih adulteranata, a test za detekciju kanabinoida najosjetljiviji, posebice u pogledu lažno negativnih rezultata. Najsnažniji adulterant koji je utjecao na rezultate testa a nije promijenio fiziološke parametre urina je alkoholni ocat. Osim soka od limuna, alkoholni ocat dodan u urin proizveo je najveći broj negativnih ishoda testiranja. Može se zaključiti da značajan broj adulteranata utječe na rezultate testa i da je moguće da neki od njih, poput alkoholnoga octa, mogu proći neopaženo pri kontroli valjanosti uzorka. Ovi nalazi upozoravaju na važnost strogo kontroliranog okruženja za uzorkovanje urina kako bi se spriječile manipulacije

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

Ispitivanje uticaja magnezijuma, cinka i bakra na izlučivanje kadmijuma kod kunića

Cadmium (Cd) is today one of the most significant metal poisons, both in the area of professional as well as of eco toxicology. In the organism, cadmium has a harmful effect on the kidneys, liver, bones, testicles, etc., and, based on evidence in humans that it causes lung carcinoma, it has been placed in the first group of carcinogens. In spite of numerous data in literature on the harmful effects of cadmium, the interactions between cadmium and bioelements as a significant mechanism for cadmium toxicity have still not been sufficiently explained. Since the data so far point to a positive effect of supplementation with certain bioelements regarding toxicity and cadmium content in the organism, the objective of this work was to investigate the effect of increased simultaneous intake of magnesium, zinc and copper on urinary elimination of cadmium in rabbits exposed to cadmium. Rabbits were divided into two groups: Cd group - for a period of 28 days the animals received per os 10 mg Cd/kg b.m/day and Cd+(Mg+Zn+Cu) group - 10 mg Cd/kg b.m. + 40 mg Mg/kg b.m, 20 mg Zn/kg b.m. and 10 mg Cu/kg b.m/day. Daily urine was collected on days 0, 10, 15, 17, 19, 21, 23, 25, and 28 of the experiment. Following decomposition of urine samples with the help of concentrated HNO3 and HClO4 (4:1), the metal concentration was determined using the method of atomic absorption spectrophotometry. Supplementation with magnesium, zinc and copper did not result in significant changes in the elimination of cadmium through urine in animals poisoned with cadmium, in comparison with the animals that were administered only cadmium, while the concentration of all three applied bioelements in urine was significantly increased. It can be concluded that the simultaneous administration of increased doses of zinc, copper and magnesium does not have a positive effect on the elimination of cadmium in conditions when rabbits are poisoned with cadmium.Kadmijum (Cd) je danas jedan od najznačajnijih metalnih otrova, kako u oblasti profesionalne tako i ekotoksikologije. U organizmu, Cd štetno deluje na bubrege, jetru, kosti, testise itd., a na osnovu dokaza da na ljudima izaziva karcinom pluća svrstan je i u I grupu karcinogena. I pored brojnih literaturnih podataka o štetnim efektima Cd, interakcije kadmijuma i bioelemenata kao značajnog mehanizma toksičnosti kadmijuma, još uvek nisu dovoljno razjašnjene. S obzirom na to da dosadašnji podaci ukazuju na pozitivan efekat suplementacije pojedinim bioelementima na toksičnost i sadržaj kadmijuma u organizmu, cilj ovog rada je bio da se ispita uticaj povećanog istovremenog unosa magnezijuma, cinka i bakra na urinarnu eliminaciju kadmijuma kod kunića izloženih kadmijumu. Kunići su bili podeljeni u dve grupe: I Cd grupa - životinje su 28 dana primale per os 10 mg Cd/kg t.m/dan i II grupa - Cd+ (Mg+Zn+Cu): 10 mg Cd/kg t.m. + 40 mg Mg/kg t.m, 20 mg Zn/kg t.m. i 10 mg Cu/kg t.m/dan. Dnevni urin je sakupljan 0, 10, 15, 17, 19, 21, 23, 25. i 28. dana eksperimenta. Nakon razaranja uzoraka urina uz pomoć koncentrovanih HNO3 i HClO4 (4:1), koncentracija metala je određena metodom atomske apsorpcione spektrofotometrije. Suplementacija magnezijumom, cinkom i bakrom nije dovela do značajne promene u eliminaciji kadmijuma urinom kod životinja trovanih kadmijumom u poređenju sa životinjama koje su primale samo kadmijum, dok je koncentracija sva tri primenjena bioelementa u urinu bila značajno povećana. Može se zaključiti da istovremeno davanje povećanih doza cinka, bakra i magnezijuma ne ispoljava pozitivan efekat na eliminaciju kadmijuma u uslovima trovanja kunića kadmijumom

Obrazovanje zdravstvenih radnika u Srbiji: procena i mere za unapređenje kvaliteta kroz praksu, interprofesionalno obrazovanje i razvoj nastavničkih kompetencija

Background/Aim. Health professions education is facing emerging issues. A comprehensive situation analysis was performed among academic staff, healthcare practitioners, and healthcare science students to address and respond to new trends. The aim of the study was to investigate the attitude, perception, and the recognized needs towards experiential education (EE), interprofessional education (IPE), and teaching competencies development (TCD). The critical evaluation of the existing quality standards for further quality improvement in health professions education in Serbia was provided. Methods. The survey on EE, IPE, and TCD was conducted within the Reinforcement of the Framework for Experiential Education in Serbia (ReFEEHS) project, co-funded by the Erasmus+ program of the European Comission at four Serbian universities (the University of Belgrade, the University of Kragujevac, the University of Niš, and the University of Novi Sad). Four task groups were appointed to perform a desk review of the existing curricula, recommendations, and practices within each of the four health professions education (Medicine, Pharmacy, Dentistry, and Nursing) in Serbia and assess the level of compliance with relevant educational policies and practices in the European Union . Results. A total of 1,507 respondents completed the survey. A highly expressed positive attitude was found towards EE, IPE, and TCD among all the respondents. The majority of the respondents (> 70%) shared that EE should be organized in real-life practice and involve students’ work under the supervision of a qualified supervisor, as well as interactions with patients and healthcare professionals. About 90% of the respondents supported the inclusion of IPE teaching activities into EE, with 77% of students expressing high motivation to attend those classes, whereas 93% of academic staff was eager to deliver and teach joint IPE subjects. Only 20% of academic staff has already attended some TCD program, while 75% recognized the need for its organization. Moreover, 90% of healthcare practitioners have recognized that mentors/clinical supervisors also need additional skills for effective mentoring work within health science education. Based on the survey results, recommendations for improvement were given within three educational fields, healthcare science curricula, professional practice (traineeship), teaching staff, and regulations. Conclusion. The results derived from the survey served as a starting but also a vital point for higher education improvement in Serbia. All interested parties – academia, students, healthcare professionals, and regulatory bodies should collaborate on achieving improved, contemporary, and transformative health professions education.Uvod/Cilj. Obrazovanje u oblasti zdravstva suočava se sa novim pitanjima. Da bi se odgovorilo na savremene zahteve obrazovanja zdravstvenih radnika, sprovedena je sveobuhvatna situaciona analiza. Cilj istraživanja bio je da se istraže stavovi, percepcije i prepoznaju potrebe akademske zajednice, zdravstvenih radnika i studenata zdravstvenih profesija, u odnosu prema učenju u realnom radnom okruženju, tj. nastavi u praksi (experiential education - EE), interprofesionalnom obrazovanju (interprofessional education - IPE) i unapređenju nastavničkih kompetencija (teaching competencies development - TCD). Radi daljeg unapređenja u oblasti obrazovanja, za zdravstvene radnike u Srbiji obezbeđeno je kritičko vrednovanje postojećih standarda kvaliteta. Metode. Istraživanje je sprovedeno u okviru projekta Reinforcement of the Framework for Experiential Education in Serbia (ReFEEHS), ko-finansiranog od strane Erasmus+ programa Evropske komisije na četiri univerziteta u Republici Srbiji (Univerzitet u Beogradu, Univerzitet u Kragujevcu, Univerzitet u Nišu i Univerzitet u Novom Sadu). Određene su četiri radne grupe, sa zadatkom razmatranja trenutnih kurikuluma, preporuka i prakse u okviru svake zdravstvene profesije (medicina, farmacija, stomatologija, sestrinstvo), kao i procene njihove usklađenosti sa relevantnim preporukama i praksom u obrazovanju u Evropskoj uniji. Rezultati. U istraživanju je učestvovalo 1 507 ispitanika. Pozitivan stav prema EE, IPE i TCD zabeležen je među svim ispitanicima. Više od 70% ispitanika iskazalo je stav o tome da je potrebno organizovati EE, tj. nastavu u praksi u realnom radnom okruženju, što bi podrazumevalo stručni rad studenata pod nadzorom kompetentnog mentora, ali i interakciju sa bolesnicima, kao i interakciju sa zdravstvenim radnicima. Oko 90% ispitanika podržalo je uključivanje IPE nastavnih aktivnosti u kurikulume; 77% studenata izrazilo je motivisanost da prisustvuju zajedničkim predmetima, dok je čak 93% nastavnika i saradnika iskazalo volju da učestvuju u kreiranju i podučavanju nastavnih jedinica u okviru IPE. Prethodno je samo 20% nastavnika/saradnika pohađalo neki TCD program, dok je čak 75% prepoznalo potrebu za organizacijom tih programa; 90% zdravstvenih radnika smatralo je da su za efikasan mentorski rad u obrazovanju u oblasti zdravstvene nauke potrebne dodatne veštine mentora/kliničkih supervizora. Na osnovu sagledanih rezultata upitnika, date su preporuke za unapređenje visokog obrazovanja u okviru tri obrazovne oblasti budućih zdravstvenih radnika, koje se odnose na kurikulum, studentsku stručnu praksu/klinički staž, nastavno osoblje i regulatorne aspekte. Zaključak. Dobijeni rezultati korišćeni su kao polazna, ali veoma značajna tačka za unapređenje visokog obrazovanja u Srbiji. Sve zainteresovane strane - akademska zajednica, studenti, zdravstveni radnici i regulatorna tela, treba da sarađuju u cilju postizanja unapređenog i savremenog obrazovanja zdravstvenih radnika