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Introduction Diabetes mellitus (DM), a well-established independent risk factor for coronary heart disease, is currently the most serious threat to humanity health worldwide due to its alarming increase in prevalence [...], Endothelial dysfunction and the related increase in reactive oxygen species (ROS) production are important events in the pathophysiology of diabetes mellitus (DM). Methylene blue (MB) has been systematically investigated for its protective effects against refractory hypotension and mitochondrial dysfunction. We have previously demonstrated that MB improved mitochondrial respiration and partially decreased oxidative stress in diabetic rat hearts. The present study was aimed to investigate whether MB modulates vascular function and ROS production in thoracic aortic rings isolated from rats with streptozotocin-induced DM (after 4 weeks of hyperglycemia). The effects of MB (0.1 [micro]M, 30 min ex vivo incubation) on vascular reactivity in organ chamber (phenylephrine-induced contraction, acetylcholine-induced relaxation) and [H.sub.2][O.sub.2] production (assessed by ferrous iron xylenol orange oxidation assay) were investigated in vascular preparations with intact endothelium and after denudation. DM elicited a significant alteration of vascular function: increased contractility to phenylephrine, attenuation of acetylcholinedependent relaxation, and augmented [H.sub.2][O.sub.2] generation. Ex vivo incubation with MB partially reversed all these changes (by approximately 70%) in vascular segments with intact endothelial layer (but not in denuded vessels). In conclusion, MB might be useful in alleviating endothelial dysfunction and mitigating endothelial oxidative stress, observations that clearly require further investigation in the setting of cardiometabolic disease. Key words: methylene blue, endothelial dysfunction, oxidative stress, experimental diabetes. Le dysfonctionnement endothelial et l&apos;augmentation de la production de derives reactifs de l&apos;oxygene (ou ROS pour <>) qui y est liee constituent des evenements importants dans la physiopathologie du diabete sucre (DS). Le bleu de methylene (BM) a ete etudie systematiquement pour ses effets protecteurs contre l&apos;hypotension refractaire et le dysfonctionnement mitochondrial. Nous avons montre precedemment que le BM permet d&apos;ameliorer la respiration mitochondriale ainsi que de faire diminuer partiellement le stress oxydatif dans le creur du rat atteint de DS. Les presents travaux visaient a etudier si le BM permet de moduler la fonction vasculaire et la production de ROS dans des anneaux d&apos;aortes thoraciques isolees de rats atteints de DS provoque par la streptozotocine (apres 4 semaines d&apos;hyperglycemie). Nous avons etudie les effets du BM (a 0,1 [micro]M, incubation ex vivo de 30 minutes) sur la reactivite vasculaire en chambre d&apos;organes (contraction provoquee par la phenylephrine, relaxation provoquee par l&apos;acetylcholine) et la production de [H.sub.2][O.sub.2] (evaluee a l&apos;aide du test d&apos;oxydation de fer ferreux en presence de xylenol orange) dans des preparations vasculaires dont l&apos;endothelium etait intact et apres denudation. Le DS a entraine une degradation notable de la fonction vasculaire : augmentation de la contractilite en reaction a la phenylephrine, attenuation de la relaxation dependante de l&apos;acetylcholine et augmentation de la production d&apos;[H.sub.2][O.sub.2]. L&apos;incubation ex vivo avec du BM a permis d&apos;inverser partiellement toutes ces variations (d&apos;environ 70 %) dans les segments vasculaires dotes d&apos;une couche endotheliale intacte (mais pas dans les vaisseaux denudes). En conclusion, le BM pourrait etre utile en vue d&apos;attenuer le dysfonctionnement endothelial et le stress oxydatif endothelial. Ce sont des observations qui demanderaient clairement des recherches plus poussees dans un contexte de maladie cardiometabolique. [Traduit par la Redaction] Mots-cles: bleu de methylene, dysfonctionnement endothelial, stress oxydatif, diabete experimental.

Introduction Coronary heart disease is the leading cause of mortality due to myocardial infarction and of morbidity due to heart failure. In the past decades, mitochondrial dysfunction has emerged as [...], Diabetic cardiomyopathy has been systematically associated with compromised mitochondrial energetics and increased generation of reactive oxygen species (ROS) that underlie its progression to heart failure. Methylene blue is a redox drug with reported protective effects mainly on brain mitochondria. The purpose of the present study was to characterize the effects of acute administration of methylene blue on mitochondrial respiration, [H.sub.2][O.sub.2] production, and calcium sensitivity in rat heart mitochondria isolated from healthy and 2 months (streptozotocin-induced) diabetic rats. Mitochondrial respiratory function was assessed by high-resolution respirometry. [H.sub.2][O.sub.2] production and calcium retention capacity were measured spectrofluorimetrically. The addition of methylene blue (0.1 [micro]mol x [L.sup.-1]) elicited an increase in oxygen consumption of mitochondria energized with complex I and II substrates in both normal and diseased mitochondria. Interestingly, methylene blue elicited a significant increase in [H.sub.2][O.sub.2] release in the presence of complex I substrates (glutamate and malate), but had an opposite effect in mitochondria energized with complex II substrate (succinate). No changes in the calcium retention capacity of healthy or diabetic mitochondria were found in the presence of methylene blue. In conclusion, in cardiac mitochondria isolated from diabetic and nondiabetic rat hearts, methylene blue improved respiratory function and elicited a dichotomic, substrate-dependent effect on ROS production. Key words: rat heart mitochondria, diabetes mellitus, methylene blue, mitochondrial respiration, reactive oxygen species. On a systematiquement associe la cardiomyopathie diabetique a une atteinte de la fonction energetique des mitochondries et a une augmentation de la production de derives reactifs de l&apos;oxygene (ROS), lesquels phenomenes sous-tendent son evolution vers l&apos;insuffisance cardiaque. Le bleu de methylene est un medicament redox avec des effets protecteurs rapportes principalement dans les mitochondries cerebrales. La presente etude a ete realisee en vue de caracteriser les effets de l&apos;administration aigue de bleu de methylene sur la respiration mitochondriale, la production de [H.sub.2][O.sub.2] et la sensibilite au calcium dans des mitochondries isolees de creurs de rats sains ou diabetiques de 2 mois (diabete provoque par la streptozotocine). Nous avons evalue la fonction respiratoire mitochondriale a l&apos;aide de la respirometrie a haute resolution. Nous avons mesure la production de peroxyde d&apos;hydrogene et la capacite de retention calcique a l&apos;aide de la spectrofluorometrie. L&apos;ajout de bleu de methylene (0,1 [micro]mol x [L.sup.-1]) a provoque une augmentation de la consommation d&apos;oxygene par des mitochondries energisees par des substrats des complexes I et II, et ce, chez les mitochondries normales et lesees a la fois. Il est interessant de noter que le bleu de methylene a provoque une augmentation marquee de la liberation de [H.sub.2][O.sub.2] en presence de substrats du complexe I (glutamate et malate), mais qu&apos;il avait un effet contraire dans les mitochondries energisees avec le substrat du complexe II (succinate). En presence de bleu de methylene, nous n&apos;avons observe aucune variation de la capacite de retention du calcium dans les mitochondries saines ou diabetiques. En conclusion, dans les mitochondries isolees de creurs de rats sains ou diabetiques, le bleu de methylene permettait d&apos;ameliorer la fonction respiratoire et provoquait un effet dichotomique et dependant du substrat sur la production de ROS. [Traduit par la Redaction] Mots-cles : mitochondrie de creur de rat, diabete sucre, bleu de methylene, respiration mitochondriale, derives reactifs de l&apos;oxygene.

Introduction Coronary heart disease remains the leading cause of mortality and morbidity due to heart failure worldwide (Go et al. 2014). In the past decades, mitochondria have emerged as key [...], Monoamine oxidases (MAOs) have recently emerged as important mitochondrial sources of oxidative stress in the cardiovascular system. Generation of reactive oxygen species during the brief episodes of ischemic preconditioning (IPC) is responsible for the cardioprotection at reperfusion. The aim of this study was to assess the effects of two MAO inhibitors (clorgyline and pargyline) on the IPC-related protection in isolated rat hearts. Animals subjected to 30 min global ischemia and 120 min reperfusion were assigned to the following groups: (i) Control, no additional intervention; (ii) IPC, 3 cycles of 5 min ischemia and 5 min reperfusion before the index ischemia; (iii) IPC-clorgyline, IPC protocol bracketed for 5 min with clorgyline (50 µmol/L); (iv) IPC-pargyline, IPC protocol bracketed for 5 min with pargyline (0.5 mmol/L). The postischemic functional recovery was assessed by the left ventricular developed pressure (LVDP) and the indices of contractility (+dLVP/dt max) and relaxation (-dLVP/dt max). Infarct size (IS) was quantified by TTC staining. In both genders, IPC significantly improved functional recovery that was further enhanced in the presence of either clorgyline or pargyline. IS reduction was comparable among all the preconditioned groups, regardless of the presence of MAO inhibitors. In isolated rat hearts, acute inhibition of MAOs potentiates the IPC-induced postischemic functional recovery without interfering with the anti-necrotic protection. Key words: monoamine oxidase, MAO inhibitors, ischemic preconditioning, isolated rat heart. Les monoamine oxydases (MAOs) se sont recemment revelees d'importantes sources de stress oxydatif mitochondrial dans le systeme cardiovasculaire. La generation de derives reactifs de l'oxygene au cours d'episodes brefs de preconditionnement ischemique (PCI) est responsable de la cardioprotection lors de la reperfusion. La presente etude visait a evaluer les effets de deux inhibiteurs de la MAO (la clorgyline et la pargyline) sur le PCI dans un modele de caeur isole de rat. Nous avons soumis les animaux a une ischemie globale de 30 min, puis a une reperfusion de 120 min et les avons repartis dans les groupes suivants: (i) temoin (aucune intervention supplementaire), (ii) PCI (3 cycles de 5 min d'ischemie suivis de 5 min de reperfusion avant l'ischemie de reference), (iii) PCI-clorgyline (protocole de PCI alternant avec l'administration de clorgyline a 50 µmol/L pendant 5 min) et (iv) PCI-pargyline (protocole de PCI alternant avec l'administration de pargyline a 0,5 mmol/L pendant 5 min). Nous avons ensuite evalue le retablissement fonctionnel post-ischemique en mesurant la pression ventriculaire gauche developpee (PVGD) ainsi que les indices de contractilite (+dP/dt max) et de relaxation (-dP/dt max). Enfin, nous avons quantifie la taille de l'infarctus par la technique de coloration au TTC. Chez les animaux des deux sexes, le PCI a permis d'ameliorer clairement le retablissement fonctionnel qui a ete accentue en presence de clorgyline ou de pargyline. Par ailleurs, la diminution de la taille de l'infarctus etait comparable dans tous les groupes soumis au preconditionnement, et ce, sans egard a la presence d'inhibiteurs de la MAO. En somme, dans les caeurs isoles de rat, l'administration a court terme d'inhibiteurs de la MAO potentialise la recuperation fonctionnelle post-ischemique induite par le PCI, sans nuire a la protection contre la necrose. [Traduit par la Redaction] Mots-cles: monoamine-oxydase, inhibiteurs de la MAO, preconditionnement ischemique, caeur isole de rat.

5-methylcytosine (5mC) is a key epigenetic mark which influences gene expression and phenotype. In vertebrates, this epigenetic mark is sensitive to Cd exposure, but there is no information linking such an event with changes in global 5mC levels in terrestrial gastropods despite their importance as excellentecotoxicological bioindicators of metal contamination. Therefore, we first evaluated total 5mC content in DNA of the hepatopancreas of adult Cantareus aspersus with the aim to determine whether this epigenetic mark is responsive to Cd exposure. The experiment was conducted under laboratory conditions and involved a continuous exposure, multiple dose- and time-point (14, 28, and 56 days) study design. Hepatopancreas cadmium levels were measured using Flame Atomic Absorption Spectrometry and the percentage of 5-mC in samples using an ELISA-based colorimetric assay. Snail death rates were also assessed. Our results, for the first time, reveal the presence of 5mC in C. aspersus and provide evidence for Cd-induced changes in global 5mC levels in DNA of gastropods and mollusks. Although less sensitive than tissue accumulation, DNA methylation levels responded in a dose- and time-dependent manner to dietary cadmium, with exposure dose having a much stronger effect than exposure duration. An obvious trend of increasing 5mC levels was observed starting at 28 days of exposure to the second highest dose and this trend persisted at the two highest treatments for close to one month, when the experiment was terminated after 56 days. Moreover, a strong association was identified between Cd concentrations in the hepatopancreas and DNA methylation levels in this organ. These data indicate an overall trend towards DNA hypermethylation with elevated Cd exposure. No consistent lethal effect was observed, irrespective of time point and Cd-dosage. Overall, our findings suggest that the total 5mC content in DNA of the hepatopancreas of land snails is responsive to sublethal Cd exposure and give new insights into invertebrate environmental epigenetics.

Introduction Ageing is the inexorable process of gradual decrease in body functional capacity, for which the underlying molecular basis remains largely elusive. It has been widely accepted that the functional [...], It is widely recognized that mitochondrial dysfunction is a key component of the multifactorial process of ageing. The effects of age on individual components of mitochondrial function vary across species and strains. In this study we investigated the oxygen consumption, the mitochondrial membrane potential (Δψ), the sensitivity of mitochondrial permeability transition pore (mPTP) to calcium overload, and the production of reactive oxygen species (ROS) in heart mitochondria isolated from old compared with adult healthy Sprague-Dawley rats. Respirometry studies and Δψ measurements were performed with an Oxygraph-2k equipped with a tetraphenylphosphonium electrode. ROS production and calcium retention capacity were measured spectrofluorimetrically. Our results show an important decline for all bioenergetic parameters for both complex I and complex II supported-respiration, a decreased Δψ in mitochondria energized with complex I substrates, and an increased mitochondrial ROS production in the old compared with the adult group. Mitochondrial sensitivity to [Ca.sup.2+]-induced mPTP opening was also increased in the old compared with the adult animals. Moreover, the protective effect of cyclosporine A on mPTP opening was significantly reduced in the old group. We conclude that healthy ageing is associated with a decrease in heart mitochondria function in Sprague-Dawley rats. Key words: ageing, oxidative phosphorylation, reactive oxygen species, calcium retention capacity. Il est generalement reconnu que la dysfonction mitochondriale est une composante cle du processus multifactoriel qu'est le vieillissement. Les effets de l'age sur les composantes individuelles de la fonction mitochondriale varient en fonction des especes et des souches. Dans cette etude, nous avons examine et compare la consommation d'oxygene, le potentiel membranaire (Δψ), la sensibilite du pore de transition de permeabilite mitochondriale (PTPM) a une surcharge en calcium et la production d'especes reactives d'oxygene (ERO) des mitochondries du coeur isolees de rats en sante Sprague-Dawley (SD) adultes vs ages. Les etudes en respirometrie et les mesures du Δψ ont ete realisees avec un Oxygraph-2k equipe d'une electrode au tetraphenylphosphonium. La production d'ERO et la capacite de retention du calcium (CRC) ont ete mesurees par spectrofluorometrie. Nos resultats ont montre un declin important de tous les parametres bioenergetiques de la respiration au niveau des complexes I et II, une diminution du Δψ dans les mitochondries tirant leur energie des substrats du complexe I et un accroissement de la production d'ERO mitochondriales chez le groupe age comparativement au groupe adulte. La sensibilite du PTPM a l'ouverture induite par le calcium etait aussi accrue chez les animaux ages comparativement aux adultes. De plus, l'effet protecteur de la cyclosporine A sur l'ouverture du PTPM etait significativement reduite chez le groupe age. En conclusion, le vieillissement normal est associe a la reduction de la fonction des mitochondries du coeur chez les rats Sprague-Dawley. [Traduit par la Redaction] Mots-cles: vieillissement, phosphorylation oxydative, especes reactives d'oxygene, capacite de retention du calcium.

OBJECTIVES AND BACKGROUND Sublethal generation of reactive oxygen species (ROS) mediates the cardioprotective effect of ischemic preconditioning (IPC). However, the sources of ROS are partially elucidated. Monoamine oxidases (MAOs) A and B are mitochondrial enzymes representing a constant source of hydrogen peroxide (H2O2) in the heart. AIM This study was purported to assess the effects of two MAO inhibitors (clorgyline for MAO-A and pargyline for MAO-B) on the IPC-related protection in isolated rat hearts. MATERIALS AND METHODS Hearts subjected to 30 min global ischemia and 120 min reperfusion were assigned to the following groups: (i) Control - no intervention; ii) IPC - 3 cycles of 5 min ischemia/5 min reperfusion before global ischemia, (iii) IPC-Clorgyline - IPC plus clorgyline (50 µM/l, iv) IPCPargyline – IPC plus pargyline (0.5 mM/l). Post-ischemic functional recovery was assessed by measuring the left ventricular developed pressure (LVDP) and the indices of contractility (+dLVP/dtmax) and relaxation (- dLVP/dtmax). Infarct size (IS) was quantified by triphenyl-tetrazolium chloride staining. RESULTS Patients IPC significantly improved functional recovery that was further enhanced in the presence of either clorgyline or pargyline. IS reduction was comparable among all preconditioned groups. CONCLUSIONS In isolated rat hearts MAO inhibition enhanced the IPC-related functional recovery without interfering with infarct size reduction. REFERENCES 1. Muntean DM, Sturza A, Danila MD, Borza C, Duicu OM, Mornos0 C. The role of mitochondrial reactive oxygen species in cardiovascular injury and protective strategies. Oxid Med Cell Longev. 2016; Article ID 8470394. 2. Edmondson DE. Hydrogen peroxide produced by mitochondrial monoamine oxidase catalysis: biological implications. Curr Pharm Design. 2014;20:155-160. 3. Di Lisa F, Canton M, Carpi A, Kaludercic N, Menabo R, Menazza S. Mitochondrial injury and protection in ischemic pre- and postconditioning. Antioxid Redox Signal. 2011;14:881-891.

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Modulation of mitochondrial function in the settings of ischemia/reperfusion (I/R) injury represents a major therapeutic target of pharmacological agents able to reduce the cell death at reperfusion. The aim of this study was to compare the effects of acute administration of magnesium orotate (MO), MgCl2, orotic acid (OA) with those elicited by the classic mitochondrial permeability transition pore (mPTP) desensitizer, cyclosporine A (CsA) in two experimental settings. The pharmacological agents were administered: i) at reoxygenation in neonatal cardiomyocytes subjected to 4 hours simulated ischemia followed by 2 hours reoxygenation with the assessment of cell viability and reactive oxygen species (ROS) generation, and ii) at reperfusion in isolated rat hearts subjected to 30 min of global ischemia and 120 min of reperfusion. In the latter model myocardial functional recovery (at 30 min of reperfusion) and infarct size (at 120 min of reperfusion) were assessed. In a separate group of animals, mitochondria were isolated at 15 min of reperfusion after global ischemia, and mitochondrial respiration, ROS production (Amplex Red) and calcium retention capacity were assessed. Acute administration of 5 mM MO, MgCl2, OA or 1 μM CsA in neonatal cardiomyocytes subjected to simulated ischemia/reoxygenation enhanced cell survival; these protective effects were not associated with changes in ROS production during reoxygenation. During the postischemic reperfusion in rat hearts and isolated mitochondria, respectively, MO, MgCl2 (5 mM both) and CsA (0.2 μmol/L), but not OA: (i) induced an important recovery of left ventricular end-diastolic pressure (LVDP), and decreased infarct size vs. controls, and (ii) protected against loss of outer mitochondrial membrane integrity and subsequent deficits in electron transport and also inhibited the calcium triggered-mPTP opening. In particular, the effects of MO were comparable to the ones elicited by CsA. In conclusion, acute administration of magnesium orotate in high concentration during the postischemic reperfusion is associated with cardioprotective effects via modulation of mitochondria functions. [ABSTRACT FROM AUTHOR]

Whether cardioprotective strategies are still effective in elderly is a matter of debate. The present study was purported to assess the persistence of the protective effects of pharmacological agents targeting mitochondria in old Sprague-Dawley rats. Methods. To this aim isolated hearts from old (20-24 mo) and adult (4-6 mo) animals subjected to a protocol of 30 min ischemia/120 min reperfusion were randomized to receive throughout the postischemic reperfusion: no intervention, magnesium orotate (MO), magnesium chloride (MgCl2) and orotic acid (OA) at 5 x 10 (-3) M. The effects on functional recovery and infarct size were compared to the ones elicited by CsA at 2 x 10(-7) M, the classic mitochondrial permeability transition pore (mPTP) desensitizer. In a second group of experiments, cardiac mitochondria were isolated at 15 min of postischemic reperfusion for the assessment of respiratory function and calcium retention capacity. Results. Acute administration of MO and MgCl2 (but not of OA) during the postischemic reperfusion was associated with a significant recovery of left ventricular developed pressure and infarct size reduction in both adult and old animals. Cardioprotection was comparable to the one elicited by CsA. In mitochondria isolated from the old animals and energized with complex I (but not complex II) substrates, all the pharmacological agents were equally effective in protecting against the loss of outer mitochondrial membrane integrity, albeit to a lesser degree compared to adult mitochondria. Also, MO, MgCl2, and CsA (but not OA) prevented the calcium triggered-opening of mPTP. Conclusion. High dose of magnesium containing-pharmacological agents and cyclosporine A were effective in protecting heart mitochondria in aged rodents during the postischemic reperfusion.This work was supported by a grant of the Ministry of National Education, CNCS–UEFISCDI, project number PN-II-ID-PCE-2012-4-0512. [ABSTRACT FROM PUBLISHER]

Pravastatin (PR), a HMG-CoA reductase inhibitor, and nicotinic acid (NA), a water-soluble vitamin B, are widely prescribed lipid-modifying drugs. Controversy exists about the effects of these agents on mitochondrial function. The present study was purported to characterize the acute effects of PR and NA (10 μM) on mitochondrial respiration in H9c2 cardiomyoblasts. Mitochondrial respiration of permeabilized cells was assessed by high-resolution respirometry using the Oxygraph-2k in the presence of complex I and complex II substrates (glutamate/malate and succinate) and, of the successive addition of the electron transport chain inhibitors: rotenone, oligomycin, FCCP, and antimycin A, respectively. Our preliminary data show an inhibitory effect on mitochondrial respiration in the presence of pravastatin, whereas nicotinic acid stimulated respiration in H9c2 cardiomyoblasts. [ABSTRACT FROM AUTHOR]

Controversy exists over the role of the active form of vitamin D, 1,25-dihydroxycholecalciferol, in the cardiovascular system in the presence of diabetes mellitus. The present study were purported to investigate the protective effect of 1,25-vitamin D3 on endothelial function in diabetic rats. To this aim, male Sprague-Dawley rats (8 weeks old) were injected with streptozotocin (STZ, 50 mg/kg, single injection intraperitoneally) to induce diabetes. After one month of hyperglycemia, thoracic aorta was removed and used for organ bath experiments. Endothelial function was evaluated in aortic rings isolated from diabetic and control animals, after 24 hours incubation in the absence or presence of 1,25-cholecalciferol (0.1 μM). Endothelial-dependent relaxation to acetylcholine was significantly reduced in diabetic as compared with normal rats. Vitamin D significantly improved vascular function in diabetic rat aortic segments. In conclusion, in vitro administration of vitamin D alleviated diabetes-induced endothelial dysfunction in experimental setting. [ABSTRACT FROM AUTHOR]

Mitochondrial dysfunction and reactive oxygen species (ROS) generation are critical events in the pathophysiology of ageing and chronic disorders, in the past years modulation of mitochondrial function becoming thus an important therapeutic target. Pharmacological agents able to improve mitochondrial function and/or to decrease ROS production are nowadays highly investigated. Among them an emerging compound is methylene blue (MB), a tricyclic phenothiazine, used for more than a century to treat a variety of disorders. Due to its redox activity, MB has a high affinity for numerous tissue oxidases, including those within mitochondria. Here we briefly review the use of MB in medicine and biology and discuss its beneficial effects as a mild-redox agent able to prevent mitochondria-driven disorders. Methylene blue has a significant potential to improve mitochondrial metabolism and represents a promising therapeutic approach in several conditions associated with mitochondrial dysfunction. [ABSTRACT FROM AUTHOR]

Unlabelled: The aim of the present work was to standardize the working methodology for assessing the bioenergetic profile of H9c2 cardiomyoblasts cells, with reference to the optimization of cell seeding number and the establishment of favorable concentrations for the classic modulators of mitochondrial respiratory function, in particular the one of a classical uncoupler, FCCP., Material and Methods: The extracellular flux analyzer (XF, Seahorse Bioscience) is a novel high-throughput instrument able to monitor the metabolism of living cells by simultaneously measuring mitochondrial respiration and glycolysis. The in vitro platform will be further used to better understand the pathophysiology and the unrecognized side effects of drugs currently used in the therapy of major cardiovascular diseases., Conclusions: In the long run, characterization of novel pharmacological agents' effects on other cell lines, including tumoral ones, will be also considered.