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Tobacco smoking remains without a doubt one of the leading causes of premature death worldwide. In combination with conventional protocols for smoking cessation, e-cigarettes have been proposed as a useful tool to quit smoking. Advertised as almost free of toxic effects, e-cigarettes have rapidly increased their popularity, becoming a sought-after device, especially among young people. Recently some health concerns about e-cigarette consumption are being raised. It is well known that they can release several toxic compounds, some of which are carcinogenic to humans, and emerging results are now outlining the risks related to the onset of respiratory and cardiovascular diseases and even cancer. The present review shows the emerging evidence about the role of technical components of the devices, the e-liquid composition as well as customization by consumers. The primary topics we discuss are the main toxicological aspects associated with e-cigarette consumption, focusing on the molecular pathways involved. Here it will be shown how exposure to e-cigarette aerosol induces stress/mitochondrial toxicity, DNA breaks/fragmentation following the same pathological pathways triggered by tobacco smoke, including the deregulation of molecular signalling axis associated with cancer progression and cell migration. Risk to fertility and pregnancy, as well as cardiovascular risk associated with e-cigarette use, have also been reported., This work was supported by a grant from the Italian Ministry of Education, University and Research. S.G., PhD fellowship grants were awarded from the Italian Ministry of Education, University and Research. L.R. and F.V. postdoctoral fellowship grant was cofounded by D.C., M.P., S.C., and P.R.; I.C-C. was supported by a postdoctoral fellowship from the Andalusian Government Ministry of Economy, Knowledge, Business, and University (DOC_00587/2020).

Phytochemicals have attracted attention in the oncological field because they are biologically friendly and have relevant pharmacological activities. Thanks to the intense and unique spicy aroma, garlic is one of the most used plants for cooking. Its consumption is correlated to health beneficial effects towards several chronic diseases, such as cancer, mainly attributable to allicin, a bioactive sulfur compound stored in different plant parts in a precursor form. The objective of this review is to present and critically discuss the chemistry and biosynthesis of allicin, its pharmacokinetic profile, its anticancer mechanisms and molecular targets, and its selectivity towards tumor cells. The research carried out so far revealed that allicin suppresses the growth of different types of tumors. In particular, it targets many signaling pathways associated with cancer development. Future research directions are also outlined to further characterize this promising natural product.

La Food and Drug Administration (FDA) ha recentemente classificato la sigaretta elettronica a tabacco riscaldato IQOS come prodotto del tabacco a rischio modificato (MRTP), incoraggiando nei consumatori la percezione che il dispositivo sia sicuro per la salute. Ad oggi, tuttavia, gli studi indipendenti non sono ancora sufficienti per delineare un profilo tossicologico adeguato di questo nuovo device e l’Organizzazione Mondiale della Sanità (OMS) invita a considerare i nuovi dispositivi elettronici per il rilascio di nicotina (DERN) come potenzialmente pericolosi per la salute, esortando la comunità medica a non contemplarli come possibili supporti nei percorsi da disassuefazione da fumo di tabacco. Inoltre, anche se in concentrazioni minori rispetto alle sigarette convenzionali (CC), l’“aerosol” prodotto da IQOS presenta nitrosammine specifiche del tabacco (TSNA), aldeidi tossiche (formaldeide, acetaldeide, acroleina), idrocarburi policiclici aromatici (PAH) e monossido di carbonio. Il presente studio si è posto l’obiettivo di studiare il potenziale impatto che l’aerosol di IQOS possa avere sul fegato, un organo target secondario, utilizzando un modello di esposizione in vivo. Ratti maschi Sprague Dawley sono stati esposti per 4 settimane all’aerosol di IQOS e, al termine, sono state effettuate analisi per la misura dei radicali liberi, del danno ossidativo e dell’attività antiossidante. Inoltre, è stata indagata la potenzialità di modulare l’attività degli enzimi del drugmetabolism e quella mitocondriale. Infine, è stato condotto uno studio di lipidomica. I risultati hanno evidenziato come IQOS provochi un incremento delle specie reattive dell'ossigeno (ROS) e dello stress ossidativo. Le riserve cellulari di glutatione totale (GSH) sono state fortemente influenzate dall’esposizione che ne ha causato una riduzione statisticamente significativa. D'altra parte, l’attività catalitica degli enzimi antiossidanti GSH-dipendenti non ha mostrato variazioni significative. Catalasi e xantina ossidasi sono risultate incrementate nel gruppo degli esposti. Inoltre, varie monoossigenasi citocromo P450-dipendenti (CYP2B1/2, CYP1A1, CYP2A1, CYP2E1) sono state indotte dall'esposizione all’aerosol di IQOS, mentre gli enzimi di fase II hanno mostrato un pattern di attività più complesso: mentre la glutatione S-transferasi (GST) è stata leggermente ridotta nel gruppo esposto, l'uridina 5'-difosfo-glucuronosil transferasi (UDPGT) è risultata marcatamente indotta. L'attività della catena respiratoria è rimasta inalterata, mentre il contenuto di mitocondri epatici è aumentato nei ratti esposti. Inoltre, l’esposizione a IQOS ha influenzato significativamente il profilo lipidico epatico. Infine, analizzando l’espressione di alcune MAP-chinasi tipicamente attivate da esposizione a fumo di CC, si è osservato come il rapporto pp38/p38 mostri un aumento statisticamente significativo nel gruppo esposto rispetto a quello di controllo, mentre il fattore di trascrizione nucleare eritroide-2 (Nrf2) è stato influenzato negativamente. Nel loro insieme, i risultati ottenuti indicano che i dispositivi IQOS potrebbero essere pericolosi non solo per gli organi bersaglio primari, ma influenzare in modo significativo anche la funzionalità epatica. Tali risultati, se pur preliminari, supportano l’atteggiamento precauzionale imposto dall’OMS nei confronti di tali dispositivi.

The Food and Drug Administration (FDA) has recently classified the electronic cigarette (e-cig) with heat-not-burn technology IQOS as a modified risk tobacco product (MRTP), thus increasing the perception of its safety. To date, independent studies do not outline a clear toxicological profile of these devices and the World Health Organization (WHO) considers new electronic nicotine delivery systems (ENDSs) as potentially harmful. Furthermore, the “aerosol” generated by IQOS contains a large number of harmful substances such as tobacco specific nitrosamines (TSNA), toxic aldehydes, polycyclic aromatic hydrocarbons (PAH) and carbon monoxide, although in a lower concentration than conventional cigarettes (CCs). This study aimed to investigate the putative impact of IQOS aerosol on the liver, a secondary target, in an in vivo model. Male Sprague Dawley rats were exposed, whole body, for 4 weeks to IQOS aerosol and free radicals’ production, oxidative damage and antioxidant activity were analysed. Additionally, drug metabolism and mitochondrial enzymatic activity as well as lipidic profile were investigated. The results obtained indicates that IQOS devices may not only be harmful to primary target organs, such as the the lungs, but also affect liver function. The results, even if preliminary, support the precautionary attitude imposed by WHO towards these devices.

Recently, the Food and Drug Administration authorized the marketing of IQOS Tobacco Heating System as a Modified Risk Tobacco Product based on an electronic heat-not-burn technology that intends to reduce the risk, due to depressed concentration of various harmful constituents typical of traditional tobacco smoke. However, its lower health risks to consumers have yet to be determined. Sprague-Dawley rats were exposed in a whole-body mode to IQOS aerosol for 4 weeks. We studied the ultrastructural changes in trachea and lung parenchyma of rats exposed to IQOS stick mainstream and tissue pro-inflammatory markers. We investigated the reactive oxygen species amount along with the markers of tissue and DNA oxidative damage. Moreover, we tested the putative genotoxicity of IQOS mainstream through Ames and alkaline Comet mutagenicity assays. Here, we observed how IQOS mainstream leads to severe remodelling of smaller and largest rat airways. We demonstrated that IQOS mainstream induces lung enzymes that activate carcinogens, increases tissue reactive radical concentration; promotes oxidative DNA breaks and gene level DNA damage; and stimulates mitogen activated protein kinase pathway which is involved in the conventional tobacco smoke- induced cancer progression. Our findings confirm that IQOS mainstream contains pyrolysis and thermogenic degradation by-products, the same harmful constituents of traditional cigarette smoke, and, for the first time, we show that it causes grave lung damage and promotes factors that increase cancer risk in the animal model.

BACKGROUND: To date, electronic cigarettes (e-cigarettes, e-cigs) have been used world- wide without people knowing the risks they were exposed. Recently, several scientific stud- ies have pointed out how e-cigs can generate carcinogens and how the exposure can lead to genomic damage through inhibiting of DNA repair or disrupting the redox homeostasis. However, some research underlined how this phenomenon can be avoided by setting the device to a low-voltage regimen. To test this feasibility, we show the effects of e-cig vapour generated from a low-voltage device filled with a nicotine-free liquid on rat testicular functions. METHODS: We investigated the chemical analysis of the e-cig vapour, the relative tes- tis weight through the gonadosomatic index (GSI), the testicular androgenic enzymes activ- ities, the testicular marker enzymes, the oxida- tive stress markers and the antioxidant and de- toxifying enzymes. Moreover, we analyzed the xanthine oxidase and the lipoxygenase activity, the cytochrome P450-linked activities and we performed the alkaline DNA unwinding test. RESULTS: The chemical analysis revealed the presence of carbonyls, such as formaldehyde, acetaldehyde and acrolein. Rats exposed re- ported a lower relative testis weight and high- er levels of lactate dehydrogenase (LDH) as tis- sue damage marker, along with an impairment of 3 βhydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD) and glucose-6-phosphate dehydrogenase (G6P- DH) as key enzymes in the steroidogenesis path- way. The pro-oxidative environment was con- firmed by the higher amount of reactive oxygen species (ROS), the development of lipid peroxi- dation and protein carbonylation, as well as from the disruption of antioxidant capability. Finally, we observed a higher rate of DNA unwinding in white blood cell line and boosted lipoxygen- ase-linked activity, a tumour promotion marker. CONCLUSIONS: Our results show that even with the setting of the device in weak condi- tion, exposure to e-cig vapours could alter the function of the gonads in male vapers.

Introduction Recently, the Food and Drug Administration authorized the marketing of IQOS Tobacco Heating System as a Modified Risk Tobacco Product based on an electronic heat-not-burn technology that purports to reduce the risk. Methods Sprague-Dawley rats were exposed in a whole-body mode to IQOS aerosol for 4 weeks. We performed the chemical characterization of IQOS mainstream and we studied the ultrastructural changes in trachea and lung parenchyma of rats exposed to IQOS stick mainstream and tissue pro-inflammatory markers. We investigated the reactive oxygen species amount along with the markers of tissue and DNA oxidative damage. Moreover, we tested the putative genotoxicity of IQOS mainstream through Ames and alkaline Comet mutagenicity assays. Results Here, we identified irritating and carcinogenic compounds including aldehydes and polycyclic aromatic hydrocarbons in the IQOS mainstream as sign of incomplete combustion and degradation of tobacco, that lead to severe remodelling of smaller and largest rat airways. We demonstrated that IQOS mainstream induces lung enzymes that activate carcinogens, increases tissue reactive radical concentration; promotes oxidative DNA breaks and gene level DNA damage; and stimulates mitogen activated protein kinase pathway which is involved in the conventional tobacco smoke-induced cancer progression. Conclusions Collectively, our findings reveal that IQOS causes grave lung damage and promotes factors that increase cancer risk. Implications IQOS has been proposed as a safer alternative to conventional cigarettes, due to depressed concentration of various harmful constituents typical of traditional tobacco smoke. However, its lower health risks to consumers have yet to be determined. Our findings confirm that IQOS mainstream contains pyrolysis and thermogenic degradation by-products, the same harmful constituents of traditional cigarette smoke, and, for the first time, we show that it causes grave lung damage and promotes factors that increase cancer risk in the animal model.

In virtù delle sue eccellenti proprietà antiossidanti negli studi di laboratorio, la vitamina E (VE) è stata per lungo tempo considerata come un dei più promettenti agenti chemiopreventivi di origine naturale. Tuttavia, le evidenze epidemiologiche più recenti non confermano tale beneficio su larga scala. Inoltre, alcuni trial randomizzati su pazienti sani al momento dell’arruolamento, hanno addirittura evidenziato un rischio d’incidenza di cancro alla prostata (CP) maggiore nel gruppo d’intervento in cui la VE è stata somministrata. È noto come la VE possa indurre alcune isoforme del CYP450 (CYP’s) sia in modelli in vitro che in vivo. L’up-regulation del sistema monoosigenasico P450 dipendente è associato ad un aumento significativo di specie reattive dell’ossigeno (ROS). Si è ipotizzato quindi che la VE potesse agire, in determinate condizioni, come un agente proossidante mediante l’induzione di alcune isoforme CYP’s. Somministrando VE a ratti Sprague-Dawley giornalmente per 7 o 14 giorni consecutivi ai dosaggi di 100 o 200 mg/kg p.c., è stato osservato una significativa induzione di alcuni CYP’s, a livello epatico. I livelli aumentati di ROS, di marcatori di stress ossidativo congiuntamente alla riduzione dell’attività di alcuni enzimi antiossidanti come catalasi (CAT) e NADPH:chinone reduttasi (NQ01), indicano come la VE possa compromettere l’omeostasi redox epatica. Anche a livello prostatico si osserva un incremento marcato dell’espressione genica di alcuni CYP’s, quindi dell’attività catalitica associata. L’aumento della concentrazione di ROS, in linea con marcatori di danno ossidativo proteico e lipidico, è stato infine confermato anche nella prostata. Utilizzando un modello in vitro basato sull’esposizione di cellule di prostata umana RWPE-1, è stato osservato, ancora una volta, una marcata induzione di alcuni CYP’s, incluse le isoforme responsabili dell’attivazione degli Idrocarburi Policiclici Aromatici (IPA), accompagnata da un significativo aumento di ROS. Inoltre, il pathway pro-infiammatorio COX-2-dipendente risulta incrementato, caratteristica che si osserva in diverse forme di CP. In vitro, i test di genotossicià hanno rilevato un’aumentata frequenza dei micronuclei (MN), inoltre lo studio della trasformazione cellulare mediante l’uso di cellule BALB/c 3T3 preincubate con VE e successivamente esposte a benzo[a]pirene, ha mostrato un significativo incremento della frequenza di trasformazione. Il presente studio dimostra come la VE possa, in determinate condizioni, agire come cocancerogeno tramite un meccanismo CYP-dipendente, causando un aumento della bioattivazione e dei ROS con conseguente effetto pro-infiammatorio, danno al DNA e promozione della trasformazione cellulare. Tali risultati possono contribuire a spiegare l’aumento dell’incidenza del CP in pazienti che hanno assunto VE a scopo chemiopreventivo.

Di recente, la Food and Drug Administration (FDA) ha autorizzato la commercializzazione dei nuovi dispositivi per il rilascio di nicotina (DERN) a riscaldamento del tabacco, come prodotti del tabacco a rischio modificato (MRTP). Si tratta di sigarette elettroniche di ultima generazione che sfruttano la tecnologia “heat-not-burn”, che scalda il tabacco evitando il processo di combustione. Distribuiti con il nome commerciale di IQOS, e concepiti allo scopo di ridurre i rischi per la salute legati al consumo dei prodotti tradizionali del tabacco, tali dispositivi hanno riscosso un notevole successo soprattutto tra i più giovani. Tuttavia, la letteratura è scevra di lavori indipendenti che ne permettano una adeguata valutazione tossicologica. Il presente studio si è posto l’obiettivo di rilevare diverse classi di composti tossici derivanti dalla degradazione termica del tabacco e le modificazioni ultrastrutturali a livello di trachea e del parenchima polmonare in ratti maschi Sprague-Dawley, a seguito di un’esposizione sub-cronica all’ “aerosol” prodotto da IQOS. È stata misurata la concentrazione di specie reattive dell'ossigeno (ROS) nel tessuto polmonare, quindi l’espressione di marcatori di danno ossidativo a livello cellulare e del DNA. Inoltre, sono stati eseguiti i test di mutagenesi di Ames su urine e Comet assay su leucociti. Nell’”aerosol” sono stati identificati composti irritanti e cancerogeni tra cui aldeidi e idrocarburi policiclici aromatici che indicano come il tabacco abbia subito una combustione incompleta. A livello delle vie aeree è stato osservato un marcato rimodellamento tissutale negli animali esposti. Inoltre, è stata riportata una significativa induzione di alcune isoforme del citocromo P450 (CYP1A1, CYP2A1/2, CYP2B1/2, CYP2E1.), una maggiore concentrazione di ROS a livello polmonare a cui segue un aumento dei marcatori di danno ossidativo al DNA, un incremento dell’espressione di alcune citochine pro-infiammatorie e l’attivazione delle principali MAP-chinasi attivate da mediatori dell’infiammazione e stress ossidativo. Infine, i test di mutagenesi eseguiti hanno mostrato come l’esposizione all’aerosol prodotto da IQOS determini un danno al DNA. Nel complesso, i risultati mostrano come IQOS possa indurre danni significativi alle vie aeree e promuovere fattori che aumentano il rischio di cancro.

Background: Designed to deliver nicotine avoiding tobacco combustion, electronic cigarettes (e-cigs) are aggressively promoted as safer alternative to tobacco products and a putative approach to smoking cessation. However, to date the risk-benefit ratio remains largely unknown and recent investigations have questioned their safety. Here we presented our recent in vivo studies on the toxicological aspects associated to the electronic delivery systems. Methods: Male Sprague Dawley rats were whole-body exposed to the vapour generated by the devices. The e-cig was equipped with different coils (1.5 and 0.25 Ohm) and the Voltage applied ranged from 3.5 to 5.5 V in order to explore the role of these parameters on toxicological outcomes. The treatment was carried out loading the device with e-liquid containing or not 18 mg/mL (1 mL/day) of nicotine. E-cig treated animals were subjected to the procedure for 3 h/day for 28 days. The levels of O2, N2, and CO2 were monitored by GC/MS to establish safe O2/N2 and CO2/ O2 ratios in the cages. Furthermore, the experiments were repeated considering also “non-liquid devices” with alternative heating systems able to heat tobacco. Results: When e-cig was run at high voltage (5.5 V; 18 mg/mL nicotine) we recorded the release of hazardous carbonyl compounds (such as formaldehyde, acetaldehyde and acrolein). The animals showed a powerful booster effect on phase-I carcinogen-bioactivating enzymes, an impairment of the antioxidant and phase II catalysts, related to the increment of oxygen free radical (ROS) production, and DNA oxidation through 8-hydroxy-2′-deoxyguanosine yield in lung tissue. Insights into the genotoxicity also emerged from the extensive DNA damage in leukocytes measured by the comet assay, the increment in the percentage of immature micronucleated reticulocytes and the increased number of revertants in the urine. Lowering the voltage at 3.5 V and loading the device with a nicotine-free liquid, we still found aldehydes in the mainstream, and noteworthy, their amount enhanced moving form nicotine to nicotine-free liquids. Interestingly, the aldehydes increased as the resistance decreased from 1.5 to 0.25 Ohm, and the frames from electron microscope confirmed a disorganization of alveolar and bronchial epithelium more marked in the 0.25 Ohm group. Finally, since changes in liver parameters including ALT and hepatocellular vacuolization in rats exposed to the vapour from “non-liquid e-cigs” were reported, we added a breakthrough finding on treated animals showing a disruption of the redox homeostasis, with higher levels of ROS, and increased markers of oxidative injuries such as lipid peroxidation and protein carbonylation, combined with an impairment of the enzymatic and non-enzymatic antioxidant defence systems. Conclusion: As a whole, our studies show how electronic delivery devices can trigger detrimental phenomena typically induced by conventional cigarettes.

Ellagitannins may have a beneficial impact in cardiovascular diseases. The aim of the study was to evaluate the effect of high-fat diet (HFD) and the efficacy of Castanea sativa Mill. bark extract (ENC) on cardiac and vascular parameters. Rats were fed with regular diet, (RD, n = 15), HFD (n = 15), RD + ENC (20 mg/kg/day by gavage, n = 15), and HFD + ENC (same dose, n = 15) and the effects on body weight, biochemical serum parameters, and inflammatory cytokines determined. Cardiac functional parameters and aorta contractility were also assessed on isolated atria and aorta. Results showed that ENC reduced weight gain and serum lipids induced by HFD. In in vitro assays, HFD decreased the contraction force of left atrium, increased right atrium chronotropy, and decreased aorta K+ -induced contraction; ENC induced transient positive inotropic and negative chronotropic effects on isolated atria from RD and HFD rats and a spasmolytic effect on aorta. In ex vivo experiments, ENC reverted inotropic and chronotropic changes induced by HFD and enhanced Nifedipine effect more on aorta than on heart. In conclusion, ENC restores metabolic dysfunction and cardiac cholinergic muscarinic receptor function, and exerts spasmolytic effect on aorta in HFD rats, highlighting its potential as nutraceutical tool in obesity.

The electronic cigarettes (e-cigarettes, e-cigs) have become the most sought-after alternative to the traditional cigarettes, partly due to the widespread perception of safety. However, the high temperature reached by e-cig solutions can generate toxic compounds, some of which are listed as known human carcinogens. To evaluate the impact of e-cig aerosol on rat brain lipid profile, twenty male Sprague Dawley rats were exposed to 11 cycles/day (E-cig group), to consume 1 mL/day of e-liquid, for 5 days/week up to 8 weeks. Ten rats were sacrificed after 4 weeks (4w) and ten at the end of treatment (8w). The composition of total fatty acids, sterols and oxysterols of the lipid fraction of rat brains, was analyzed. The results of the E-cig group were compared with those of the control group (not exposed). After 8 weeks, the saturated fatty acids significantly raised up to 7.35 mg/g tissue, whereas polyunsaturated fatty acids decreased reaching 3.17 mg/g. The e-cig vaping increased both palmitic (3.43 mg/g) and stearic acids (3.82 mg/g), while a significant decrement of arachidonic (1.32 mg/g) and docosahexaenoic acids (1.00 mg/g) was found. Atherogenic (0.5) and thrombogenic (1.12) indices also increased in 8w treated animals. The e-cig aerosol significantly impacted the cholesterol homeostasis, since the latter at 8w (21.57 mg/g) was significantly lower than control (24.56 mg/g); moreover, a significant increase of 7-dehydrocholesterol (1.87 mg/g) was also denoted in e-cig group. The e-cig aerosol also reduced the oxysterol formation (19.55 μg/g) after 4 weeks of exposure, except for triol and 5α,6α-epoxycholesterol (α-EC). The principal component analysis (PCA) separated all E-cig from control groups, evidencing that oxysterols (except triol and 24(S)-hydroxycholesterol (24(S)-HC)) were inversely correlated to 7-DHC and TI. The present research revealed that e-cigs aerosol affected the lipid and cholesterol homeostasis in rat brain, which could contribute to the new occurrence of some neurodegenerative diseases.

Despite the knowledge gap regarding the risk-benefit ratio of the electronic cigarette (e-cig), its use has grown exponentially, even in teenagers. E-cig vapor contains carcinogenic compounds (eg, formaldehyde, acetaldehyde, and acrolein) and free radicals, especially reactive oxygen species (ROS) that cause toxicological effects, including DNA damage. The role of e-cig voltage customization on molecule generation has been reported, but the effects of the resistance on e-cig emissions and toxicity are unknown. Here, we show that the manipulation of e-cig resistance influences the carbonyls production from nonnicotine vapor and the oxidative and inflammatory status in a rat model. Fixing the voltage at the conventional 3.5 V, we observed that the amount of the selected aldehydes increased as the resistance decreased from 1.5 to 0.25 Ω. Under these conditions, we exposed Sprague Dawley rats to e-cig aerosol for 28 days, and we studied the pulmonary inflammation, oxidative stress, tissue damage, and blood homeostasis. We found a perturbation of the antioxidant and phase II enzymes, probably related to the increased ROS levels due to the enhanced xanthine oxidase and P450-linked monooxygenases. Furthermore, frames from scanning electron microscope showed a disorganization of alveolar and bronchial epithelium in 0.25 Ω group. Overall, various toxicological outcomes, widely recognized as smoke-related injuries, can potentially occur in e-cig consumers who use low-voltage and resistance device. Our study suggests that certain “tips for vaping safety” cannot be established, and encourages further independent investigations to help public health agencies in regulating the e-cig use.

Introduction: Electronic cigarettes (e-cigs) are devices thought to deliver nicotine in a vaping solution without tobacco combustion. Perceived as a safer alternative to conventional cigarettes, e-cigs are aggressively marketed as lifestyle-choice consumables, thanks to few regulatory restrictions. E-cigs have also gained popularity among never-smokers and teenagers, becoming an emergent public health issue. Despite the burgeoning worldwide consumption, their safety remains largely unproven and it is unknown whether these devices could cause in vivo toxicological effects contributing to cancer. Materials and methods: Male Sprague Dawley rats were randomly split into two groups: non-exposed and whole-body mode exposed (10 animals per group). Treated animals consumed 1 mL/day of e-liquid containing 18 mg/mL of nicotine. During the experiment the e-cigarette voltage was set at 5.5 (and the wattage was about 15). Animals were exposed for 4 hours/day for 5 consecutive days/week, and for 4 consecutive weeks. Results: Toxic aldehydes production, due to thermal decomposition of glycerol, were detected in e-cig aerosol. In exposed lung, we observed a significant increase in phase I CYP1A1/2, CYP2B1/2, CY2C11 and CYP3A enzymes. The overproduction of reactive oxygen species resulting from CYP induction, together with the free radicals present in the aerosol, was confirmed by means “Electron paramagnetic resonance-radical probe” technique. Notably, we observed that the lung antioxidant enzymes catalase, DT-diaphorase, superoxide dismutase and the conjugating phase II glutathione S-transferases were all markedly reduced in animal treated. In addition, we found a markedly reduced FRAP (ferric reducing antioxidant power) value in the lung of e-cig rodents. Moreover, plasma FRAP levels and measurement of carbonyl residues, as biomarkers of oxidative injury to proteins, were inversely correlated in e-cig vapour-exposed rats. We also found that 8-hydro-2′-deoxyguanosine markedly increased in the lungs of treated rats. Interestingly, e-cigs produce extensive DNA damage in leukocytes (Comet test). We also found that e-cig vapour determines an increase in the percentage of immature micronucleated reticulocytes over normal reticulocyte. Next, the urine of e-cig-exposed animals induced a dose-dependent increase in the number of S. typhimurium revertants in different strains. Discussion and conclusion: We found that e-cigs have a powerful booster effect on phase-I carcinogen-bioactivating enzymes, and increase oxygen free radical production and DNA oxidation to 8-hydroxy- 2′-deoxyguanosine. Furthermore, we found that e-cigs damage DNA not only at chromosomal level in peripheral blood, such as strand breaks in leucocytes and micronuclei formation in reticulocytes, but also at gene level such as point mutations in urine. As these detrimental phenomena are typically induced by conventional cigarettes, the erroneous belief that e-cigs are safe should be retracted and suitable measures implemented to protect public health. Our study should be seen as the starting point for further investigations designed to confirm the harmful health impact of e-cigs, particularly after long-term exposure and under different usage conditions.

Introduction The dietary supplements use has increased over the past years in the global landscape and vitamins are by far the products which record the greater appeal. Following the well-rooted idea that the vitamin E promotes health status, many of people assume it not to treat diagnosed deficiencies, but to prevent new occurrences of several diseases, including cancer. However, recent trials and meta-analysis showed no significant effects against cancer aetiology, whereas data from SELECT study reported an increased risk of prostate cancer among healthy men. To date, the mechanism underlying these observations is largely unknown. In this work, we show in in vitro and in vivo models, how vitamin E through co-mutagenic and pro-oxidant mechanisms promotes both DNA damage and cell transformation frequency. Materials and Methods In vitro models Human prostate cells RWPE-1 were exposed to α-tocopherol (100 μM) for 48 h. The gene expression of COX2, CYPA1, CYPA4, CYPB6, CYPC9, and CYP4F2 gene was studied. PGE2 were determinate by ELISA test. The measure of reactive oxygen species (ROS) was carried out by the use of the Electron Paramagnetic Resonance (EPR) radical-probe technique. Human fibroblasts IMR90 were exposed to α-tocopherol (100 μM) for 48 h then cells were fixed, stained with DAPI. Micronuclei were scored. IMR90 cultured as previously described were employed for the Immunofluorescence determination of 53BP1 and γH2AX foci. BALB/c 3T3 cells were treated with α-tocopherol (100 μM), after 48 h exposition the medium was removed and cells were incubated with benzo[a]pyrene with a final concentrations ranged from 1 µg/ml to 0.01 µg/ml. Cells were maintained in culture for 4 weeks with twice weekly medium changes, then fixed and stained. Positive transformation foci were counted. In vivo model Vitamin E was administered to Sprague-Dawley rats at doses of 100 or 200 mg/kg b.w. dose daily for 7 or 14 consecutive days. Controls received vehicle (corn oil). Prostate was resected and the CYP1A1 CYP1A2, CYP1B1 gene expression was studied along with the catalytic activity through specific probes. ROS yields was measured by EPR spectroscopy. Finally, the malonyldialdehyde (MDA) development, as a marker of lipid peroxidation as well as protein carbonylation were assessed. Results Here we show that in RWPE-1 cell line vitamin E produces an overexpression of various phase-I bioactivating cytochrome P450 (CYP) isoforms, including activators of polycyclic aromatic hydrocarbons (PAHs), such as CYPA1, CYPA4, CYPB6, CYPC9, and CYP4F2 coupled with a significant yield of radical oxygen species (ROS). These observations have been supported by an in vivo model; in vitamin E supplemented rats we reported a significant booster effect on CYP superfamily along with oxidative damage to lipids and proteins. Furthermore, we show that vitamin E causes DNA damage as exemplified through micronuclei and H2AX - 53BP1 foci based-assays. Finally, we find that the pre-exposure of BALB/c 3T3 cells with vitamin E significantly increase the transformation frequency of the PAH-prototype benzo[a]pyrene. Discussion and Conclusions Taken together, these results can contribute to explain the increased cancer risk found in the SELECT trial. Our data support the hypothesis that vitamin E could act as a co-mutagenic/co-carcinogenic agent increasing the bioactivation of chemicals joint with its pro-oxidant activity resulting in higher risk of cancer through DNA insults and epigenetic alterations. Our study point out one more time the potentially harmful effects of recommending supplementations with micronutrient as a chemopreventive strategy on a large scale.

Purpose: We previously showed the positive effects of the new antioxidant molecule bis(1-hydroxy-2,2,6,6-tetramethyl-4-piperidinyl)-decandioate (IAC) in reducing basal hyperglycaemia and relieving glucose intolerance in a diabetes model. However, the chemical properties of IAC did not allow an efficient oral administration, thus representing the main failing of that study. Here, we tested the effect of a new oral delivery system based on solid lipid microparticles (SLMs) in a diabetes mouse model. Methods: The diabetes model was induced in C57B1/6J mice using streptozotocin and nicotinamide. Only the animals that overcame the glycaemic threshold of 180 mg/dL were enrolled in the study. Diabetic animals were then randomly assigned to 4 groups (n = 9) and treated once a day for 5 consecutive weeks with IAC (50, 100, and 150 mg/kg b.w.). The control group was composed of (n = 7) healthy mice that received only the vehicle. Glucose level was weekly monitored during the treatment period and up to 3 weeks after the suspension of the treatment. Glucose tolerance and insulin-resistance test were carried out. Results: Our results showed that SLMs maintained the IAC effect in reducing basal hyperglycaemia as well as improving the insulin sensitivity and glucose tolerance. Conclusion: The present study confirms that SLMs are promising drug carriers, which allow the oral administration of IAC ensuring its therapeutic efficacy. The concrete possibility to administer IAC per os represents a significant breakthrough in the putative consideration of this multi-radical scavenger in the diabetes therapeutic approach.

The main purpose of this study was to compare the benefits of SSJ supplementation in obese rats with those achieved only by switching the alimentary regimen from high-fat (HFD) to the regular one (RD) in liver, ileum and prostate. Furthermore, changings in caecal chime microbiota were investigated. SSJ was administered to rats in combination with a RD (HFD-RD + SSJ). The switch from HFD to RD led to a weight loss of almost 9.8 g, and the total cholesterol was found to be significantly lower. In the HFD-RD + SSJ group, all values were improved compared with the HFD control, and the weight decrement was higher (-23.29 g) with respect to HFD-RD. HFD led to a widespread increment of oxidative stress (OS) markers in liver, ileum and prostate. SSJ has shown to improve the results achieved by the suspension of HFD and it has proven effective wherever the only switch in diet regimen failed.

A large number of basic researches and observational studies suggested the cancer preventive activity of vitamin E, but large-scale human intervention trials have yielded disappointing results and actually showed a higher incidence of prostate cancer although the mechanisms underlying the increased risk remain largely unknown. Here we show through in vitro and in vivo studies that vitamin E produces a marked inductive effect on carcinogen-bioactivating enzymes and a pro-oxidant status promoting both DNA damage and cell transformation frequency. First, we found that vitamin E in the human prostate epithelial RWPE-1 cell line has the remarkable ability to upregulate the expression of various phase-I activating cytochrome P450 (CYP) enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), giving rise to supraphysiological levels of reactive oxygen species. Furthermore, our rat model confirmed that vitamin E in the prostate has a powerful booster effect on CYP enzymes associated with the generation of oxidative stress, thereby favoring lipid-derived electrophile spread that covalently modifies proteins. We show that vitamin E not only causes DNA damage but also promotes cell transformation frequency induced by the PAH-prototype benzo[a]pyrene. Our findings might explain why dietary supplementation with vitamin E increases the prostate cancer risk among healthy men.

Despite the lack of knowledge of the effects of electronic cigarettes (e-cigarettes, e-cigs) on public health, they have been proposed as a part of smoking cessation efforts. Recently, several basic scientific studies have pointed out how e-cigs can generate carcinogens, such as e-cig liquid thermal degradation by-products, and how the exposure can lead to genomic damage through inhibiting DNA repair or disrupting the redox homeostasis. However, scientific studies have pointed out how e-cigs can generate carcinogens and their release could be avoided setting the device to a low-voltage regimen. To test this feasibility, we show the effects of e-cig vapour generated from a low-voltage device filled with a nicotine-free liquid on rat testicular functions. The chemical analysis revealed the presence of carbonyls, such as formaldehyde, acetaldehyde and acrolein. Rats exposed reported a lower relative testis weight and higher levels of lactate dehydrogenase (LDH) as tissue damage marker, along with an impairment of 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD) and glucose-6-phosphate dehydrogenase (G6PDH) as key enzymes in the steroidogenesis pathway. The pro-oxidative environment was confirmed by the higher amount of reactive oxygen species (ROS), the development of lipid peroxidation and protein carbonylation, as well as from the disruption of antioxidant capability. Finally, we observed a higher rate of DNA unwinding in white blood cell line and boosted lipoxygenase (LOX)-linked activity, a tumour promotion marker. Even with the device setting at weak conditions, our results if extrapolated to humans suggest that exposure to e-cig vapours might alter gonads function in male vapers.

Electronic cigarette (e-cigarette; e-cig) use has grown exponentially in recent years despite their unknown health effects. E-cig aerosols are now known to contain hazardous chemical compounds, including carbonyls and reactive oxygen species (ROS), and these compounds are directly inhaled by consumers during e-cig use. Both carbonyls and ROS are formed when the liquid comes into contact with a heating element that is housed within an e-cig's atomizer. In the present study, the effect of coil resistance (1.5 Ω and 0.25 Ω coils, to obtain a total wattage of 8 ± 2W and 40 ± 5 W, respectively) on the generation of carbonyls (formaldehyde, acetaldehyde, acrolein) and ROS was investigated. The effect of the aerosols generated by different coils on the viability of H1299 human lung carcinoma cells was also evaluated. Our results show a significant (p < 0.05) correlation between the low resistance coils and the generation of higher concentrations of the selected carbonyls and ROS in e-cig aerosols. Moreover, exposure to e-cig vapor reduced the viability of H1299 cells by up to 45.8%, and this effect was inversely related to coil resistance. Although further studies are needed to better elucidate the potential toxicity of e-cig emissions, our results suggest that these devices may expose users to hazardous compounds which, in turn, may promote chronic respiratory diseases.

The oxidative stress theory of aging has brought to the implicit expectation that oxidative stress increases with aging. Unfortunately, a broad investigation in humans is missing due to limitations of conventional oxidative stress status (OSS) analyses. Here we show that the OSS measured in peripheral blood of 247 healthy volunteers, aged 2 days-104 years, using the electron paramagnetic resonance "EPR-radical probe" technique, negatively correlated with age (-1.1 %/year; p < 0.0001) both by simple and multiple linear regression analyses and that it was only marginally affected by sex. These findings stimulate further mechanistic studies.

We previously showed that the innovative radical scavenger bis(1-hydroxy-2,2,6,6-tetramethyl-4-piperidinyl)-decandioate (IAC) improves metabolic dysfunctions in a diabetic mouse model. Here, we compared the in vivo effects of IAC with those of the anti-diabetic drugs pioglitazone (PIO) and exendin-4 (EX-4). Diabetes was induced in C57Bl/6J mice by streptozotocin and nicotinamide administration. Paralleled by healthy controls, diabetic animals (D) were randomly assigned to four groups and treated daily for 7 consecutive weeks: D+saline, ip; D+IAC 30mg/kgb.w., ip; D+PIO 10mg/kgb.w. per os; and D+EX-4, 50μg/kgb.w., ip. Our results show that IAC reduced basal hyperglycemia and improved glucose tolerance better than PIO or EX-4. Interestingly, in the heart of diabetic mice, IAC treatment normalized the increased levels of GSSG/GSH ratio and thiobarbituric acid reactive substances, indexes of oxidative stress and damage, while PIO and EX-4 were less effective. As supported by immunohistochemical data, IAC markedly prevented diabetic islet β-cell reduced density, differently from PIO and EX-4 that had only a moderate effect. Interestingly, in diabetic animals, IAC treatment enhanced the activity of pancreatic-duodenal homeobox 1 (PDX-1), an oxidative stress-sensitive transcription factor essential for maintenance of β-cell function, as evaluated by quantification of its nuclear immunostaining, whereas PIO or EX-4 treatments did not. Altogether, these observations support the improvement of the general redox balance and β-cell function induced by IAC treatment in streptozotocin-nicotinamide diabetic mice. Furthermore, in this model, the correction of diabetic alterations was better obtained by treatment with the radical scavenger IAC than with pioglitazone or exendin-4.

Geraniol is a natural monoterpene showing anti-inflammatory, antioxidant, neuroprotective and anticancer effects. No pharmacokinetic and bioavailability data on geraniol are currently available. We therefore performed a systematic study to identify the permeation properties of geraniol across intestinal cells, and its pharmacokinetics and bioavailability after intravenous and oral administration to rats. In addition, we systematically investigated the potential hepatotoxic effects of high doses of geraniol on hepatic phase I, phase II and antioxidant enzymatic activities and undertook a hematochemical analysis on mice. Permeation studies performed via HPLC evidenced geraniol permeability coefficients across an in vitro model of the human intestinal wall for apical to basolateral and basolateral to apical transport of 13.10 ± 2.3 x 10−3 and 2.1 ± 0.1 x 10−3 cm/min, respectively. After intravenous administration of geraniol to rats (50 mg/kg), its concentration in whole blood (detected via HPLC) decreased following an apparent pseudo-first order kinetics with a half-life of 12.5 ± 1.5 minutes. The absolute bioavailability values of oral formulations (50 mg/kg) of emulsified geraniol or fiber-adsorbed geraniol were 92% and 16%, respectively. Following emulsified oral administration, geraniol amounts in the cerebrospinal fluid of rats ranged between 0.72 ± 0.08 µg/mL and 2.6 ± 0.2 µg/mL within 60 minutes. Mice treated with 120 mg/kg of geraniol for four weeks showed increased anti-oxidative defenses with no signs of liver toxicity. CYP450 enzyme activities appeared only slightly affected by the high dosage of geraniol.

The electronic cigarettes (e-cigarettes, e-cigs) have become the most sought-after alternative to the traditional cigarettes, partly due to the widespread perception of safety. However, the high temperature reached by e-cig solutions can generate toxic compounds, some of which are listed as known human carcinogens. To evaluate the impact of e-cig aerosol on rat brain lipid profile, twenty male Sprague Dawley rats were exposed to 11 cycles/day (E-cig group), to consume 1 mL/day of e-liquid, for 5 days/week up to 8 weeks. Ten rats were sacrificed after 4 weeks (4w) and ten at the end of treatment (8w). The composition of total fatty acids, sterols and oxysterols of the lipid fraction of rat brains, was analyzed. The results of the E-cig group were compared with those of the control group (not exposed). After 8 weeks, the saturated fatty acids significantly raised up to 7.35 mg/g tissue, whereas polyunsaturated fatty acids decreased reaching 3.17 mg/g. The e-cig vaping increased both palmitic (3.43 mg/g) and stearic acids (3.82 mg/g), while a significant decrement of arachidonic (1.32 mg/g) and docosahexaenoic acids (1.00 mg/g) was found. Atherogenic (0.5) and thrombogenic (1.12) indices also increased in 8w treated animals. The e-cig aerosol significantly impacted the cholesterol homeostasis, since the latter at 8w (21.57 mg/g) was significantly lower than control (24.56 mg/g); moreover, a significant increase of 7-dehydrocholesterol (1.87 mg/g) was also denoted in e-cig group. The e-cig aerosol also reduced the oxysterol formation (19.55 μg/g) after 4 weeks of exposure, except for triol and 5α,6α-epoxycholesterol (α-EC). The principal component analysis (PCA) separated all E-cig from control groups, evidencing that oxysterols (except triol and 24(S)-hydroxycholesterol (24(S)-HC)) were inversely correlated to 7-DHC and TI. The present research revealed that e-cigs aerosol affected the lipid and cholesterol homeostasis in rat brain, which could contribute to the new occurrence of some neurodegenerative diseases.

In recent years, health protection by natural products has received considerable attention, and a multitude of nutraceuticals have been characterized and their use promoted. Dietary consumption of Cruciferous vegetables, rich in glucosinolates (GLs), and their myrosinase-mediated hydrolysis products isothiocyanates (ITCs), were associated with reductions in cancer risk. In this study, the chemo-preventive potential of sprout extract of Tuscan black cabbage ( Brassica oleracea L. var. acephala subvar. Laciniata L.) (TBCSE), through modulation of the xenobiotic-metabolizing apparatus and antioxidant defenses, was investigated in Sprague-Dawley rat liver. TBCSE was administered either orally or intraperitoneally, at a dose of 15 mg/kg b.w., daily for twenty-one consecutive days, in the absence or presence of exogenous myrosinase, β-thioglucoside glucohydrolase (MYR), to distinguish the effects of intact GLs and ITCs, in the context of the extract. A complex, mild modulation pattern of P450-related monooxygenases was observed, mainly regarding CYP content (up to 36% loss), NADPH cytochrome (P450) c -reductase (up to 26% loss), CYP1A1 (up to 23% loss), but no evident distinctions among the effects of the extracts containing GLs or ITCs, were noted. In contrast, significant inductions of phase-II enzymes (up to 107% for UDP-glucuronosyl-transferase, and up to 36% for glutathione S -transferase) were recorded only where the GLs to ITCs conversion had occurred. A boosting effect on catalase (up to 38%), NAD(P)H:quinone reductase (up to 70%), glutathione reductase and glutathione peroxidase (up to 10%) was also recorded, suggesting an indirect antioxidant capacity of the extracts. Overall, the general phase-I inhibition, together with the up-regulation of detoxifying phase-II and antioxidant enzymes, exerted by the TBCSE supplementation, seem to be in line with the classical chemopreventive theory, but whether the addition of exogenous MYR is relevant, still remains to be clarified. These results are in support of the potential health-promoting application of TBCSE, as a nutraceutical.

The present study aimed to evaluate the impact of Raphanus sativus cv Sango sprout juice (SSJ) administration (75 mg/kg b.w. SSJ/day) on the brain lipidomic profile (fatty acid, sterols, cholesterol oxidation) of rats (non-genetic model) subjected to a high-fat (34% crude fat) dietary regimen. The SSJ did not affect the lipid infiltration (7.7–9.3%) and the fatty acid composition of the rat brain, which was mainly composed by unsaturated fatty acids (∼ 58%); however, the high-fat diet regimen significantly halved linoleic acid (LA). The high-fat diet also decreased (21.13 mg/g) the level of brain cholesterol with respect to the regular diet (4.5% crude fat) (23.83 mg/g); however, when the diet was shifted from high-fat to a regular regimen with or without SSJ supplementation, the levels of cholesterol significantly (p < 0.05) increased up to 30.46 mg/g of brain. The main oxysterols were 24(S)-hydroxycholesterol (24(S)-HC) and β-epoxycholesterol (β-EC). The high-fat diet led to the highest cholesterol oxidation (63.1 μg/g), increasing 27-hydroxycholesterol (27-HC) infiltration (0.24 μg/g rat brain) through the blood-brain barrier (BBB) compared to the regular diet (0.13 μg/g rat brain). On the other hand, when the diet was switched from high-fat to a regular regimen with SSJ supplementation, a significant reduction of 27-HC in the rat brain was found. Although 24-HC did not significantly change (p = 0.054), an increasing trend was observed when high-fat diet was supplied. The principal component analysis (PCA) revealed that SSJ was more active in counteracting cholesterol oxidation when supplied with the high-fat diet, due to inverse correlation with 24(S)-HC and 27-HC; however, further studies are needed to better understand which is the relationship between LA and cholesterol homeostasis in rat brain.

Electronic cigarettes (e-cigs) are devices designed to deliver nicotine in a vaping solution rather than smoke and without tobacco combustion. Perceived as a safer alternative to conventional cigarettes, e-cigs are aggressively marketed as lifestyle-choice consumables, thanks to few restrictions and a lack of regulatory guidelines. E-cigs have also gained popularity among never-smokers and teenagers, becoming an emergent public health issue. Despite the burgeoning worldwide consumption of e-cigs, their safety remains largely unproven and it is unknown whether these devices cause in vivo toxicological effects that could contribute to cancer. Here we demonstrate the co-mutagenic and cancer-initiating effects of e-cig vapour in a rat lung model. We found that e-cigs have a powerful booster effect on phase-I carcinogen-bioactivating enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), and increase oxygen free radical production and DNA oxidation to 8-hydroxy-2′-deoxyguanosine. Furthermore, we found that e-cigs damage DNA not only at chromosomal level in peripheral blood, such as strand breaks in leucocytes and micronuclei formation in reticulocytes, but also at gene level such as point mutations in urine. Our results demonstrate that exposure to e-cigs could endanger human health, particularly among younger more vulnerable consumers.

The digestibility (in vitro), toxicity and metabolic effects of rapeseed (RPH) and sunflower (SPH) protein hydrolysates have been evaluated in a murine animal model. The enzyme AlcalaseVR was employed to obtain a mild enzymatic hydrolysis of rapeseed and sunflower defatted seed meals (DSM) protein isolates. Both hydrolysates showed higher in vitro digestibility than the respective DSM, presumably as a consequence of the hydrolysis process that they had undergone. In vivo, RPH and SPH were well tolerated. Body and organ weights, biochemical blood parameters from treated male mice were comparable to controls. Food intake was regular in RPH and SPH animals, suggesting a good palatability of the hydrolysates. Not relevant perturbations of the principal hepatic and renal drug metabolism enzymes were observed in RPH or SPH mice. In conclusion, protein hydrolysates from sunflower and rapeseed DSM did not determine relevant toxicological effects; therefore, they could be considered as alternative protein sources and/or food ingredients.

Despite more than 50 years of investigations into the free radical theory, the direct role of oxidative stress (OS) in aging and age-related diseases remains unproven. Little progress in identifying antioxidant drugs promoting longevity has been made, likely due to selectivity toward one or few radical species, variable efficacy in vivo, inherent pro-oxidant behavior of such drugs, or lack of synergism with metabolic redox homeostasis. Silencing the wide range of reactive free radicals has a great impact on OS-linked outcomes and age-related disorders. Here we show that an innovative, redox-active, multi-radical-scavenger catalytic drug delays the age-associated decline in physiological processes and markedly prolongs the mean lifespan of the adult freshwater annelids Aeolosoma viride by 170%. This unprecedented extension is associated with a decreased OS status. Consistently, treatment of annelids increases their natural resistance to oxygen-derived damage without affecting mitochondrial respiration or reproductive activity. Conversely, the superoxide dismutase (SOD)-mimetic EUK 134 that we selected as a positive control led to an increase in lifespan of ~50%, the same increase previously observed in nematodes. Our results show that reduction of the global network of OS has a profound impact on aging, prompting the development of a possible redox-based therapeutic intervention to counteract the progression of aging.

The bis (1-hydroxy-2,2,6,6-tetramethyl-4-piperidinyl)-decandioate (IAC), is an innovative non- radical scavenger used with success in numerous disease models such as inflammation, neurological disorders, hepatitis and diabetes. The pharmacological treatments have been performed by the intraperitoneal route of administration, representing to date, the main limit for the drug use. The aim of this study was to develop a delivery system that allows the oral administration of IAC while maintaining its therapeutic efficacy. Solid Lipid Microparticles (SLMs) containing a theoretical 18% (w/w) of IAC have been produced by the spray congealing technology; three formulations have been tested (A, B and C) using different low melting point carriers (stearic acid, Compritol® HD5ATO and carnauba wax) alone or in combination. All IAC loaded SLMs exhibited a spherical shape, encapsulation efficiency higher than 94% and particle size suitable for the oral route. Administered per os at different dosages in an inflammation rat model, all SLMs demonstrated their efficacy in reducing oedema and alleviating pain, compared to the gold standards Indomethacin and Paracetamol. These results suggested that the SLMs are an efficacious delivery system for the oral administration of IAC, potentially useful for the treatment of others diseases related to an over production of free radicals.

Over the past years, there has been a growing interest in the natural constituents of foods as a potential means of cancer control. To date, epidemiology studies seem to indicate an inverse association between regular consumption of fruit and vegetables and cancer risk. Here, the potential chemopreventive activity of the polyphenolic extract (PPE) of peach (Prunus persica L. Batsch) and of the freeze-dried fruit in toto (LFT), focusing on the modulation of xenobiotic metabolizing enzymes (XMEs) in vivo, was investigated. Rats were daily supplemented with LFT at 250 and 500 mg/kg b.w. or with the corresponding amount of PPE (2.5 and 5 mL/kg b.w., respectively) for either 7 or 14 days. While PPE treatment resulted in a widespread phase-I inactivation, a complex modulation pattern with drastic decreases (7α-testosterone hydroxylase, pentoxyresorufin O-dealkylase (PROD)), coupled with marked up-regulations of ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O-demethylase (MROD) after LFT administration, was seen. A notable down-regulation (over 50%) following LFT or PPE treatment for the phase-II enzymes was also recorded. The observed remarkable changes in XMEs, if reproduced in humans, might have public health implications. These data suggest caution in promoting peach fruit (mono-diet) consumption or its polyphenolic extract in the field of chemoprevention.

Fruit and vegetables (FV) have long been considered a panacea against major chronic diseases, including cancer. However, there is no convincing epidemiological, clinical or experimental evidence supporting FV chemopreventive ability. A daily mono-supplementation of lyophilized onion, tomato, peach, black grape or lettuce was compared with the daily combined administration of the same FV (5 a day-like diet). Ten days post-treatment, the phase-I/II xenobiotic metabolizing and antioxidant enzyme activities, protein and mRNA levels were investigated. As a marker of oxidative stress, the level of hydroperoxides was measured in rat serum samples. Here we show that a blend of FV orally administered to rats not only potentially manipulates metabolism but also disrupts systemic oxidative homeostasis. A daily combination of the five servings remarkably down-regulates the catalytic activity, protein and mRNA levels of a cohort of hepatic metabolizing enzymes, suggesting a possible depressed clearance upon exposure to ubiquitous carcinogens. Strikingly, we observed an impairment of antioxidant enzymes with a boost in systemic hydroperoxide levels. Our study identifies new potential factors of cancer risk connected with the persistent consumption of fixed servings of FV, suggesting that dietary guidance should rely on a "daily diversification" of FV.

Drinking water (DW) disinfection represents a milestone of the past century, thanks to its efficacy in the reduction of risks of epidemic forms by water micro-organisms. Nevertheless, such process generates disinfection by-products (DBPs), some of which are genotoxic both in animals and in humans and carcinogenic in animals. At present, chlorination is one of the most employed strategies but the toxicological effects of several classes of DBPs are unknown. In this investigation, a multidisciplinary approach foreseeing the chemical analysis of chlorinated DW samples and the study of its effects on mixed function oxidases (MFOs) belonging to the superfamily of cytochrome P450-linked monooxygenases of Cyprinus carpio hepatopancreas, was employed. The experimental samples derived from aquifers of two Italian towns (plant 1, river water and plant 2, spring water) were obtained immediately after the disinfection (A) and along the network (R1). Animals treated with plant 1 DW-processed fractions showed a general CYP-associated MFO induction. By contrast, in plant 2, a complex modulation pattern was achieved, with a general up-regulation for the point A and a marked MFO inactivation in the R1 group, particularly for the testosterone metabolism. Together, the toxicity and co-carcinogenicity (i.e. unremitting over-generation of free radicals and increased bioactivation capability) of DW linked to the recorded metabolic manipulation, suggests that a prolonged exposure to chlorine-derived disinfectants may produce adverse health effects.

Aims A large meta-analysis of randomized clinical trials has seriously questioned chemoprevention based on vitamins including vitamin E (VE), and an increased risk for cancer among long-term users was actually seen. However, the mechanism underlying these findings still remain unknown. To clarify the mechanism, in an in vivo model we studied the putative disruption of redox homeostasis and the perturbation of carcinogen metabolizing enzymes determined by VE. Main methods Male Sprague–Dawley rats were treated ip with either 100 or 200 mg/kg b.w. daily for 7 or 14 consecutive days. Controls received vehicle only. Cytochrome P450 (CYP) content, CYP-reductase, CYP-linked monooxygenases, as well as phase-II and the antioxidant enzymes catalase and NAD(P)H:quinone reductase were investigated in both liver and kidney. Free radical species in tissue subcellular preparations were measured by electronic paramagnetic resonance (EPR) spectroscopy coupled to a radical probe technique. Key findings No substantial changes of hepatic xenobiotic metabolism enzymes were determined by VE. Conversely, a powerful booster effect of various renal phase-I carcinogen bioactivating enzymes at both dosages and observational times was recorded. While no relevant changes of post-oxidative phase-II reactions were found in the liver, a significant inactivating effect was caused by VE in renal tissues. Antioxidant enzymes were found mainly downregulated by the treatment. In the kidney, a marked free radical over-generation linked to CYP induction was observed. Significance This study proved that VE acts as a co-carcinogen and pro-oxidant agent. Such epigenetic mechanisms may contribute to explain the harmful outcomes observed in humans.