Cysteine proteases obtained from the stem of pineapple or papaya latex, bromelain and papain, respectively, exhibit a broad spectrum of beneficial effects on human health. However, their effects on gut microbiota composition or dose-manner effects on the intestinal integrity of healthy tissue have not been evaluated. In this study, C57BL/6 young, healthy mice were fed bromelain or papain in a dose of 1 mg per animal/day for three consecutive days, followed by the assessment of digestive protein capacity, intestinal morphology and gut microbiota composition. Furthermore, a human reconstructed 3D tissue model EpiIntestinal (SMI-100) was used to study the effects of 1, 0.1 and 10 mg/mL doses of each enzyme on tissue integrity and mucosal permeability using TEER measurements and passage of Lucifer Yellow marker from the apical to the basolateral side of the mucosa. The results indicated that fruit proteases have the potential to modulate gut microbiota with decreasing abundance of Proteobacteria and increasing beneficial Akkermansia muciniphila. The enhancement of pancreatic trypsin was observed in bromelain and papain supplementation, while bromelain also increased the thickness of the ileal mucosa. Furthermore, an in vitro study showed a dose-dependent interruption in epithelial integrity, which resulted in increased paracellular permeability by the highest doses of enzymes. These findings define bromelain and papain as promising enzymatic supplementation for controlled enhancement of paracellular uptake when needed, together with beneficial effects on the gut microbiota.
- Publikační typ
- časopisecké články MeSH
The aim of the study was toxicological testing of an innovative and efficient antimicrobial agent based on photoactive phthalocyanine (Pc) derivative. A promising Aluminium phthalocyanine (AlPc) with efficient and stable antimicrobial effects was subjected to a battery of toxicological tests to avoid local and systemic toxicity hazard. In compliance with the current European legislation restricting the use of experimental animals, the methods comprised exclusively in vitro procedures based on cellular and tissue models of human origin or mimicking human tissues. The battery of toxicological tests to identify local toxicity included skin corrosion/irritation, eye irritation, and phototoxicity. The basic systemic toxicity tests included acute toxicity, skin sensitization, genotoxicity, and endocrine disruption. The results showed that AlPc induced skin and eye irritation, exhibited borderline sensitization potential and mutagenic potential in one test strain of the Ames test, which was not confirmed in the chromosome aberration test. The AlPc was found to be phototoxic. The results from the cytotoxicity test designed for acute oral toxicity estimation were not conclusive, the acute toxicity potential has to be determined by conventional tests in vivo. Regarding endocrine disruption, no agonistic activity of the AlPc on human estrogen receptor α, nor human androgen receptor was observed. The skin penetration/absorption test revealed that the AlPc has not penetrated into the dermis and receptor fluid, confirming no risk of systemic exposure via the bloodstream.
- MeSH
- alfa receptor estrogenů metabolismus MeSH
- androgenní receptory metabolismus MeSH
- antiinfekční látky farmakokinetika toxicita MeSH
- chorioalantoická membrána krevní zásobení účinky léků MeSH
- dráždivé látky farmakokinetika toxicita MeSH
- fotochemické procesy MeSH
- indoly farmakokinetika toxicita MeSH
- kožní absorpce MeSH
- kultivované buňky MeSH
- kuřecí embryo MeSH
- kůže účinky léků metabolismus MeSH
- lidé MeSH
- lymfocyty účinky léků MeSH
- myši inbrední BALB C MeSH
- oči účinky léků MeSH
- poškození DNA MeSH
- prasata MeSH
- testy toxicity MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The study was focused on assessment of potential health risks of paper-based food contact materials (FCMs) in a step-wise approach using three toxicological bioassays in vitro and chemical analyses of migrating contaminants. 3T3 NRU cytotoxicity test showed high sensitivity to detect basal toxicity of FCMs extracts and served as a first-line test for selection of samples for further testing. The reconstructed human intestine model EpiIntestinal showed more realistic tissue response than cell culture monolayer and higher resistance despite prolonged exposure to the selected 6 samples, i.e. negligible decrease of viability and intestinal penetration, nevertheless an increase of IL-8 after exposure to black printed sample extract. Yeast based assays identified weak agonistic/antagonostic activity to human androgen receptor of the black printed sample. In accordance with the biological effects, the targeted LC and GC analytical methods confirmed the presence of high amounts of phthalates, photoinitiators and PAHs that could justify the hazard of the black printed sample. Heavily printed uncoated FCMs are recognized not to be suitable for direct contact with food. The selected bioassays and chemical analyses might be useful tools to detect targeted biological effects of xenobiotics suspected to contribute to human exposure from food.
- MeSH
- androgenní receptory genetika metabolismus MeSH
- biotest MeSH
- buňky BALB 3T3 MeSH
- cytokiny metabolismus MeSH
- hodnocení rizik MeSH
- intestinální absorpce MeSH
- kontaminace potravin MeSH
- kyseliny ftalové analýza toxicita MeSH
- lidé MeSH
- myši MeSH
- obaly potravin * MeSH
- papír * MeSH
- polycyklické aromatické uhlovodíky analýza toxicita MeSH
- receptory pro estrogeny genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika MeSH
- střevní sliznice účinky léků metabolismus MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Several irritants were used in the in vitro irritation medical device round robin. The objective of this study was to verify their irritation potential using the human patch test (HPT), an in vitro assay, and in vivo data. The irritants were lactic acid (LA), heptanoic acid (HA), sodium dodecyl sulfate (SDS), Genapol® X-80 (GP), and Y-4 polymer. Dilute saline and sesame seed oil (SSO) solutions of each were evaluated using a 4 and 18 h HPT and the EpiDerm™ SIT-MD RhE assay; results were then compared to existing rabbit skin irritation test data. Results from the 4 h HPT were negative in most cases except for GP and SDS, while the 18 h HPT also identified some LA, HA, and GP samples as irritants. EpiDerm™ SIT-MD correctly identified all irritants except GP in SSO due to limited solubility. Data from cutaneous rabbit irritation tests were negative, while all intracutaneous results were strongly or weakly positive except for the most dilute GP solutions. These findings indicate that EpiDerm™ SIT-MD results correlate with those from the rabbit intracutaneous test and confirm that RhE assays are suitable replacements for animals in evaluating the tissue irritation potential of medical devices.
- MeSH
- alternativy testů na zvířatech MeSH
- benchmarking MeSH
- dodecylsíran sodný toxicita MeSH
- dráždivé látky toxicita MeSH
- králíci MeSH
- kůže účinky léků MeSH
- kyselina mléčná toxicita MeSH
- kyseliny heptylové toxicita MeSH
- lidé MeSH
- náplasťové testy metody MeSH
- polyethylenglykoly toxicita MeSH
- polyvinylchlorid toxicita MeSH
- reprodukovatelnost výsledků MeSH
- testy kožní dráždivosti metody MeSH
- zdravotnické prostředky * MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cieľom práce je poskytnúť stručný prehľad o aktuálnej situácii v oblasti skríningu cytotoxického účinku a mechanizmu pôsobenia prírodných a syntetických látok využiteľných v chemoterapii nádorov. Medzi v súčasnosti najčastejšie sa vyskytujúce a najzávažnejšie ochorenia patria onkologické ochorenia, ktoré predstavujú pre človeka život ohrozujúce riziko. Vážnou komplikáciou protinádorovej terapie je neadekvátna terapeutická odpoveď, ktorá je zapríčinená rezistenciou ľudského organizmu na používané liečivá, chemoterapeutiká. A práve rezistencia je jednou z hnacích síl, ktoré nás neustále nútia vyhľadávať nové protinádorovo účinné liečivá, či už prírodného alebo syntetického pôvodu. Dnes sa bežne a rutinne používa na vyhľadávanie nových potenciálnych cytostatík primárny skríning in vitro, pri ktorom sa rôznymi metódami sleduje citlivosť nádorových buniek rastúcich in vitro na cytotoxické látky. Vlastnosti potenciálnej protinádorovej zlúčeniny sú charakterizované rôznymi parametrami, medzi ktoré patrí antiproliferačná aktivita, sledovanie štrukturálnych a funkčných zmien cytoplazmovej membrány, zmien v obsahu bunkových proteínov, resp. nukleových kyselín, zmien metabolizmu buniek, zmien bunkového cyklu, indukcia apoptózy, enzýmová aktivita (dihydrofolátreduktázy, proteináz, proteínkináz, topoizomeráz, tymidylátsyntetázy), vplyv na mitochondrie, cytoskelet bunky, telomerázová aktivita atď. Sledovaním týchto parametrov súčasne sledujeme aj mechanizmus účinku cytotoxicky účinnej látky.
The paper reviews the current approaches to cytotoxic effect screening and mode of action of natural and synthetic compounds usable in tumour chemotherapy. Oncological diseases belong to the most frequently occurring and the most serious diseases that threaten millions of human lives. A serious complication of anticancer therapy is an inadequate therapeutic answer which is caused by the resistance of the human organism to the employed drugs, chemotherapeutic agents. Therefore the resistance is one of the driving forces which constantly force us to search for new anticancer effective drugs of natural or synthetic origin. Primary screening in vitro, in which by different methods the sensitivity of cancer cells growing in vitro to cytotoxic compounds is monitored, is commonly and routinely used for searching for new potential cytostatics today. The properties of a potential anticancer compound are characterized by different parameters, which include antiproliferative activity, monitoring of structural and functional changes in the cytoplasmic membrane, changes in cell proteins and nucleic acids content, cell metabolism changes, cell cycle changes, induction of apoptosis, enzymatic activity (dihydrofolate reductase, proteinases, proteinkinases, topoisomerases, tymidylatsynthetase), effect on mitochondria, cell cytoskeleton, telomerase activity, etc. By monitoring these parameters, the mode of action of a cytotoxically effective compound can be followed up.
- MeSH
- antitumorózní látky fytogenní aplikace a dávkování škodlivé účinky MeSH
- antitumorózní látky aplikace a dávkování chemická syntéza škodlivé účinky MeSH
- apoptóza genetika MeSH
- buněčná smrt fyziologie účinky léků MeSH
- buněčný cyklus fyziologie účinky léků MeSH
- cytoskelet fyziologie účinky léků MeSH
- finanční podpora výzkumu jako téma MeSH
- lidé MeSH
- nádorová transformace buněk MeSH
- nežádoucí účinky léčiv MeSH
- plošný screening metody MeSH
- přehledová literatura jako téma MeSH
- Check Tag
- lidé MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- Publikační typ
- abstrakty MeSH
