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Oxidative stress is a key mediator of autoimmune/neurodegenerative disorders. The antioxidant/anti-inflammatory effect of a synthetic conjugated linoleic acid (CLA) mixture in MRL/MpJ-Faslpr mice (MRL/lpr), an animal model of neuropsychiatric lupus, was previously associated with the improvement of Nuclear factor-E2-related factor 2 (Nrf2) defences in the spleen and liver. However, little is known about the neuroprotective ability of CLA mixture. This study investigates the age-dependent progression of oxidative stress and the hyperactivation of redox-sensitive compensatory pathways (macroautophagy, Nrf2) in old/diseased MRL/lpr mice brains, and examines the effect produced by dietary CLA supplementation. Disrupted redox homeostasis was evidenced in the blood, liver and brain of 21- to 22-week old MRL/lpr mice (Old) compared to young animals (Y; 8- to 10-week old). This alteration was associated with significant hyperactivation of compensatory mechanisms (macroautophagy, Nrf2 and astrocyte activation) in the brains of Old mice. Five-week daily supplementation with CLA (650 mg/kg-1 body weight) of 16-week old mice (CLA+Old) diminished all the pathological hallmarks at a level comparable to Y mice or healthy controls (BALB/c). Such data demonstrated that MRL/lpr mice can serve as a valuable model for the evaluation of the effectiveness of neuroprotective drugs. Notably, the preventive effect provided by CLA supplementation against age-associated neuronal damage and hyperactivation of compensatory mechanisms suggests that the activation of an adaptive response is, at least in part, accountable for its neuroprotective ability.

Food colors, or color additives, are substances imparting color to commercial food and drinks and to a variety of non-food applications such as cosmetics, pharmaceuticals, home craft projects and medical devices. Of natural or artificial origin, their safety has been long discussed and concerns about consumer health led to testing for safety and to certification. From 1973 interferences with activity in children is generally accepted and warnings are now present on products labels. Much less attention however has been dedicated to the possible effects of these additives on natural flora and fauna and, in particular, on the aquatic ones. In our work we have tested the toxicity of 4 different commercially available food colors: one natural, cochineal red E120 and three of synthesis: Ponceau red E124, tartrazine yellow E102 and patent blue E131. Concentrations were the same suggested in labels for preparing food (650 mg/500 ml milk or cream). Toxicity was tested on Cucumis sativus (Cucurbitales), Artemia salina (Crustacea Anostraca) and Danio rerio (Actinopterygii Cyprinidae) development. Results have demonstrated that the four food colors significantly interfere with Cucumis germination and rootlets formation and that they moderately alter toracopods development in Artemia. In Danio rerio, embryos show significant alterations with pericardial oedema, hypopigmentation and anomalous development of tail and body axis. Results, though preliminary, suggest that food colors are potentially toxic to the flora and aquatic fauna and that attention should be devoted to this so far ignored consequence of our habit of artificially coloring the world around us.

Gold nanoparticles are increasingly being employed in innovative biological applications thanks to their advantages of material- and size-dependent physics and chemical interactions with the cellular systems. On the other hand, growing concern has emerged on the toxicity which would render gold-based nanoparticles harmful to cell cultures, animals, and humans. Emerging attention is focused on the interaction of gold nanoparticles with nervous system, especially regarding the ability to overcome the blood-brain barrier (BBB) which represents the major impediment to the delivery of therapeutics into the brain. We synthesized highly stable 2-mercapto-1-methylimidazole-stabilized gold-nanoparticles (AuNPs)-mmi to investigate their entry, accumulation, and toxicity in vitro (SH-SY5Y human neuroblastoma cells) and in vivo (brain of C57BL/6 mice) through optical and electron microscopy. After incubation in the cell culture medium at the lowest dose of 0.1 mg/mL the (AuNPs)-mmi nanoparticles were found compacted and recruited into endosome/lysosomes (1 h) before their fusion (2 h) and the onset of neuronal death by apoptosis (4 h) as proved by terminal-transferase-mediated dUTP nick end labeling assay and caspase-3 immunoreactivity. The ability of (AuNPs)-mmi to cross the BBB was assessed by injection in the caudal vein of C57BL/6 mice. Among different brain regions, the nanoparticles were found in the CaudatoPutamen area, mainly in the striatal neurons 4 h after injection. These neurons showed the typical hallmarks of apoptosis. Our findings provide, for the first time, the dynamic of 2-mercapto-1-methylimidazole nanogold uptake. The molecular mechanism which underlies the nanogold-driven apoptotic event is analyzed and discussed in order to take into account when designing nanomaterials to interface with biological structures. © 2014 Wiley Periodicals, Inc.

Acute or chronic alterations in energy status alter the balance between excitatory and inhibitory synaptic transmission and associated synaptic plasticity to allow for the adaptation of energy metabolism to new homeostatic requirements. The impact of such changes on endocannabinoid and cannabinoid receptor type 1 (CB1)-mediated modulation of synaptic transmission and strength is not known, despite the fact that this signaling system is an important target for the development of new drugs against obesity. We investigated whether CB1-expressing excitatory vs. inhibitory inputs to orexin-A–containing neurons in the lateral hypothalamus are altered in obesity and how this modifies endocannabinoid control of these neurons. In lean mice, these inputs are mostly excitatory. By confocal and ultrastructural microscopic analyses, we observed that in leptin-knockout (ob/ob) obese mice, and in mice with diet-induced obesity, orexinergic neurons receive predominantly inhibitory CB1-expressing inputs and overexpress the biosynthetic enzyme for the endocannabinoid 2-arachidonoylglycerol, which retrogradely inhibits synaptic transmission at CB1-expressing axon terminals. Patch-clamp recordings also showed increased CB1-sensitive inhibitory innervation of orexinergic neurons in ob/ob mice. These alterations are reversed by leptin administration, partly through activation of the mammalian target of rapamycin pathway in neuropeptide-Y-ergic neurons of the arcuate nucleus, and are accompanied by CB1-mediated enhancement of orexinergic innervation of target brain areas. We propose that enhanced inhibitory control of orexin-A neurons, and their CB1-mediated disinhibition, are a consequence of leptin signaling impairment in the arcuate nucleus. We also provide initial evidence of the participation of this phenomenon in hyperphagia and hormonal dysregulation in obesity.

We analyzed the effect of cadmium on corticotropic (ACTH) and prolactin (PRL) cells in the pituitary gland of the Podarcis sicula (P. sicula) lizard under chronic exposure to this metal. Adult lizards were given CdCl2 in drinking water at the dose of 10 µg/10 g body mass for 120 days. Light microscopy was performed after histological and immunohistochemical staining, and the effects were followed at regular time intervals up to 120 days post-treatment. We detected substantial variations in the general morphology of the pituitary: unlike the control lizards in which the gland appeared compact, the treated lizards showed a glandular tissue with dilated spaces that were more extensive at 90 and 120 days. PRL and ACTH cells showed an increase in occurrence and immunostaining intensity in treated lizards in comparison with the same cells of control animals. This cellular increase peaked for PRL at 30 days in the rostral, medial and also caudal pars distalis of the gland. ACTH cells appeared to increase markedly after 60 days of treatment in both the pars distalis and the pars intermedia. Again, at 60 days small, isolated ACTH cells were also found in the caudal pars distalis in which these cells were generally absent. However, at 120 days both these cellular types showed an occurrence, distribution and morphology similar to those observed in the control lizards. In lizards, protracted oral exposure to cadmium evidently involves an alteration of the normal morphology of the gland and an inhibitory effect of ACTH and PRL cells, since they increase in occurrence and immunostaining. Yet in time the inhibitory effect of cadmium on ACTH and PRL cells falls back and their occurrence appears similar to that of the control lizard.

Delivery of pharmacologic agents to neurons in the brain across the blood brain barrier (BBB) remains a challenging task for neuroscientists. Because of their unique properties, nanoparticles have generated a great deal of interest for their potential use in therapeutics drug delivery. We investigated the use of colloidal gold (AuNP) in this context based on its characteristic stability as a metal nanoparticle. We explored the neuronal uptake of AuNPs both in vitro using SH-SY5Y human neuroblastoma cells and in vivo using direct brain injection in mice. AuNPs have a diameter of approximately 2 nm and an absorption spectrum with a peak at 350 nm. Due to their characteristic quantum size effect, the nanogold emission shifts from green to red upon forming clusters of approximately 200 nm in diameter. The clustered particles have a broad emission ranging from 400 to 600 nm. For the in vitro studies, we incubated SH-SY5Y cells with 1x105 AuNPs for 20 min, 1, 2 and 4 hrs. AuNP internalization was observed using a Leica microscope workstation B6000 equipped with an in vivo timelapse analysis capability. We confirmed our results using transmission electron microscopy (TEM). For in vivo, studies we injected a physiological solution containing 1x106 AuNPs into the carotid arteries of anesthetized mice (n=3). The physiological solution alone was injected into control mice for comparison. Our in vitro data demonstrated that gold-particles passed the cell membrane, clustered in the cytoplasm and induced apoptosis after 4 hrs of treatment. In vivo, a strong particle-associated florescence was observed in the cauda putamen region (CPu) 90 min after injection. Fluorescence was evenly distributed in the neuronal cells but was not observed in glial cells, as demonstrated by GFAP, Iba1 and NeuN immunoistochemistry. The cytotoxic effects of AuNPs were investigated both in vitro and in vivo using caspase-3 immunoreactivity and TUNEL assays for apoptosis. Our data suggest that AuNPs cross the BBB and have useful imaging properties that rely on clustering in cells, which changes their emission spectrum from green to red. Specific properties of these nanoparticles may offer new advantages as potential pharmaco-delivery tools in treatment of neurological diseases that may benefit from specific neuronal cell targeting.

The treatment with antipiogenic and trivalent antiviral vaccines evokes in sargus' blood an increase of the neutrophilic granulocytes and an activation of the lymphocytes. By using cytochemical reactions for acid phosphatase and aliesterase, a population of lymphocytes, amounting to about 25%, which displays positive reactions has been identified. The immunocytochemical reactions used to identify the mammalian T and B-lymphocytes, such as resetting test, immune localization of CD (Cluster of differentiation) and of sig, were performed with the aim of better classifying the lymphocytes of sargus, even though the use of mammalian monoclonal CD antibodies cannot be considered a certain proof of such a presence. Moreover, cytochemical reactions have revealed the presence of lysosomal enzymes in neutrophils and the presence of peroxidase which characterize the eosinophils. Hemopoiesis takes place in the spleen and in the lymphomieloid tissue of the kidney.

Food dyes, or color additives, are chemicals added to industrial food products and in domestic cooking to improve the perceived flavor and attractiveness. Of natural and synthetic origin, their safety has been long discussed, and concern for human safety is now clearly manifested by warnings added on products labels. Limited attention, however, has been dedicated to the effects of these compounds on aquatic flora and fauna. For this reason, the toxicity of four different commercially available food dyes (cochineal red E120, Ponceau red E124, tartrazine yellow E102 and blue Patent E131) was assessed on three different model organisms, namely Cucumis sativus, Artemia salina and Danio rerio that occupy diverse positions in the trophic pyramid. The evidence collected indicates that food dyes may target several organs and functions, depending on the species. C. sativus rate of germination was increased by E102, while root/shoot ratio was ∼20% reduced by E102, E120 and E124, seed total chlorophylls and carotenoids were 15–20% increased by E120 and 131, and total antioxidant activity was ∼25% reduced by all dyes. Mortality and low mobility of A. salina nauplii were increased by up to 50% in presence of E124, E102 and E131, while the nauplii phototactic response was significantly altered by E102, E120 and E124. Two to four-fold increases in the hatching percentages at 48 h were induced by E124, E102 and E131 on D. rerio, associated with the occurrence of 20% of embryos showing developmental defects. These results demonstrated that the food dyes examined are far from being safe for the aquatic organisms as well as land organisms exposed during watering with contaminated water. The overall information obtained gives a realistic snapshot of the potential pollution risk exerted by food dyes and of the different organism' ability to overcome the stress induced by contamination.

We investigated the relationship between age and body length, and age at sexual maturity of Physeter macrocephalus individuals stranded along the Italian coast. Our molecular analysis shows that all our samples belong to the C.001.002 haplotype, shared between Atlantic and Mediterranean populations. We show that males attain sexual maturity at 10 years, similar to those from other marine areas. However, considering the same body length class, Mediterranean males are older than Atlantic ones. Our finding of a Mediterranean pregnant female of only 6.5 m in length and an assessed age of 24–26 years is particularly noteworthy, considering that females reach sexual maturity at about 9 years and 9 m of total length in other regions. Comparing our results with the literature data, we highlight the positive correlation between lifespan, adult body length and weight of males from the Mediterranean and Atlantic Ocean. Regardless of whether the relatively small size of Mediterranean specimens is a consequence of an inbreeding depression or an adaptation to less favorable trophic conditions, we recommend to closely monitor this population from a conservation perspective. In fact, its low genetic diversity likely corresponds to a relatively limited ability to respond to environmental changes compared with other populations.

This paper evaluates the combined effect of biostimulant and light quality on bioactive compound production and seedling growth of soybean (Glycine max L. Merrill) plants. Germinated seeds pre-treated with different concentrations (0.01%, 0.05%, 0.5%) of an amino acid-based biostimulant were grown for 4 days at the dark (D), white fluorescent light (FL), full-spectrum LED (FS), and red-blue (RB) light. Potential changes in the antioxidant content of sprouts were evaluated. Part of the sprouts was left to grow at FL, FS, and RB light regimes for 24 days to assess modifications in plants’ anatomical and physiological traits during the early developmental plant stage. The seed pre-treatment with all biostimulant concentrations significantly increased sprout antioxidant compounds, sugar, and protein content compared to the control (seeds treated with H2O). The positive effect on bioactive compounds was improved under FS and RB compared to D and FL light regimes. At the seedling stage, 0.05% was the only concentration of biostimulant effective in increasing the specific leaf area (SLA) and photosynthetic efficiency. Compared to FL, the growth under FS and RB light regimes significantly enhanced the beneficial effect of 0.05% on SLA and photosynthesis. This concentration led to leaf thickness increase and shoot/root ratio reduction. Our findings demonstrated that seed pre-treatment with proper biostimulant concentration in combination with specific light regimes during plant development may represent a useful means to modify the bioactive compound amount and leaf structural and photosynthetic traits.

Aluminium (Al), a metal extremely widespread in the world but not essential, is considered dangerous both for the environment and for human health1. It is also indicated as a possible etiological factor in neurodegenerative diseases2 although, its neurotoxic role is still not completely understood. In this study, zebrafish was used like model organism, since useful both for ecotoxicology studies and for exploring neurodegenerative diseases3. Adult zebrafish were exposed to 11 mg/L Al and the swimming ability and the behavioural responses were assessed at 10, 15 and 20 days of exposure. These parameters were correlated with the effects induced by the metal on the brain, evaluating the amount of Al within this organ, the oxidative stress, antioxidant-defences, changes in metabolism and neurotransmission. Furthermore, its neurotoxic role was further investigated evaluating the induced neurodegeneration and the gene-expression of markers involved in the parkinsonism. The behavioural and locomotory responses, suggest an increase in the anxiety state, especially in animals exposed to Al for 15 days, when also the neurodegeneration and accumulation of Al in the brain were evident. According to these data, even the activity of antioxidant enzymes, as superoxide dismutase, glutathione peroxidase and the metallothioneins levels increased after short-term exposures and tended to decrease or stabilize at longer times, though, the reactive oxygen species instead increased in a time-dependent trend. Finally, the expression of genes linked to Parkinsonism also was influenced by exposure to the metal, with an evident greater impact after short time of exposure. Overall, the results contribute to understand the neurotoxic mechanisms activated by Al highlighting correlations between behavioural disorders, oxidative state and neurodegenerative processes.

Aluminium (Al) water pollution is an increasing environmental problem. Accordingly, this study aimed to find out more about its toxic effects on aquatic organisms. Adult zebrafish were exposed to 11 mg/L of Al and the behavioural responses and its correlation with brain oxidative stress, antioxidant-defences, changes in metabolism and neurotransmission were assessed at 10, 15 and 20 days of exposure. The behavioural and locomotory responses, suggest an increase in the anxiety state, especially observed in animals exposed to Al for 15 days. The reactive oxygen species increased in a time-dependent trend, while the oxidative damage varied over exposure time. The activity of antioxidant enzymes, as superoxide dismutase, glutathione peroxidase and glutathione S-transferases, and the metallothioneins levels increased after short-term exposures and tended to decrease or stabilize at longer times. The results contribute to understand the toxic mechanisms activated by Al highlighting correlations like behavioural disorders and oxidative state.

Aluminium, widely present in daily life1, is reported to be linked to several neurological disorders. We evaluated the effects of AlCl3 on the terrestrial snail Eobania vermiculata, an excellent bioindicator of soil pollutants. Histochemical and lectin histo-chemical techniques were used to investigate glycopattern variation in the mucins secreted by the pedal glandular system,involved in several functions, such as adhesion, locomotion, and protection. Three groups of adult snails (mean shell size = 2.5cm; mean weight = 4.6 g) were fed daily with 4 g of lettuce soaked in a AlCl3 water solution at three different concentrations(0, 50, and 200 μM). After 30 days, animals were sacrificed, Bouin-fixed and embedded in paraffin. Sections, 6- m thick, were analysed by histochemical methods (PAS, AB pH 2.5, HID-ABpH 2.5) and lectin binding experiments (PNA, SBA, WGA, LTA,UEA-I, AAA, SNA, MAA II, ConA). Controls were positive toPAS, AB pH 2.5, and with HID stained mostly brown, indicatingthe presence of carboxylated and sulphated glycans. Positivity was reduced in the treated snails, where the secretion was con-centrated in the gland ducts rather than in the adenomeres. ConA(linking to mannosylated and/or glycosylated residuals) bound strongly to all the samples both in the dorsal area and the sole.SBA and WGA (linking to galactosaminylated and glycosaminy-lated/sialylated residuals, respectively) increased their binding in the dorsal area of the foot of treatments. AAA, linking to fuco-sylated residuals, decreased its binding in the treatments. In the sole, SBA-binding decreased in the treatments. In conclusion, the AlCl3 treatment affects the quali-quantitative expression of gly-cans in the foot. Physiopathological implications of changes will be investigated in further research.

E150d is a food additive used to increase palatability and impart colour to foods and drinks. Known as ‘caramel dye’, it contains 4-methylimidazole, a cytotoxic molecule for animal models and human. Recently, the growing consumption of E150d causes an increasing release of this additive into the environment, particularly in water bodies. For this reason, in this study it was assessed the toxic effect of E150d on zebrafish embryos, a conventional aquatic model organism. Six hours post fertilization embryos were treated with two different concentrations of E150d (0.3 g/L and 0.6 g/L) for 72 h and their embryonic development was studied. It emerged that this food additive induced toxic effects on hatching, survival, embryos phenotype and cardiac beat with a dose-dependent trend. Furthermore, it impaired swimming performance and induced damages in skeletal muscles and pericardial cavity. Data obtained showed the risk associated with the dispersion of E150d in water bodies suggesting that a greater attention should be paid in avoiding an unnecessary use so to preserve human and environmental health.

The effects of nitrates were analysed on the land snail Eobania vermiculata, a good bioindicator to assess the effects of certain pollutants in soil. It is known that the molluscs are very sensitive to contamination substances and can be used as sentinel organism for environmental pollution assessment. The nitrates are present in fertilizers and in food additives and their excess can not only be harmful to the environment but also dangerous for the humans. Indeed, in the mammals the nitrates are converted into nitrites and can cause a series of complications as the formation of methaemoglobin and cancers. In this study, adult organisms of E. vermiculata were exposed to soil containing 2000 mg/L of nitrates for 30 days to evaluate the stool microbiome and the histological changes at the level of the foot. Eggs of these snails were similarly treated to observe their hatching, survival and development. Histological changes were observed at level of the foot of adult snails exposed to nitrate and in their stools was evident an increase of bacteria, especially those that have a high ability to exploit nitrates and nitrogen as nutrients. Instead, the treated eggs showed changes in hatching, hypopigmentation of newborn snails and a decrease of their survival in time. The overall information obtained from these endpoints can provide important information regarding the quality of the environment. In addition, they also showed that the invertebrate organism E. vermiculata despite being a simple organism is very useful and efficient for ecotoxicological studies.

Endogenous reactive oxygen species (ROS) are by-products of the aerobic metabolism of cells and have an important signalling role as secondary messengers in various physiological processes, including cell growth and development. However, the excessive production of ROS, as well as the exposure to exogenous ROS, can cause protein oxidation, lipid peroxidation and DNA damages leading to cell injuries. ROS accumulation has been associated to the development of health disorders such as neurodegenerative and cardiovascular diseases, inflammatory bowel disease and cancer. We report that spores of strain SF185, a human isolate of Bacillus megaterium, have antioxidant activity on Caco-2 cells exposed to hydrogen peroxide and on a murine model of dextran sodium sulfate-induced oxidative stress. In both model systems spores exert a protective state due to their scavenging action: on cells, spores reduce the amount of intracellular ROS, while in vivo the pre-treatment with spores protects mice from the chemically-induced damages. Overall, our results suggest that treatment with SF185 spores prevents or reduces the damages caused by oxidative stress. The human origin of SF185, its strong antioxidant activity, and its protective effects led to propose the spore of this strain as a new probiotic for gut health.

Inflammation and oxidative stress play an important role in the pathogenesis of depressive disorders and Nuclear erythroid related factor 2 (Nrf2), a regulator of RedOx homeostasis and inflammation, is a promising target for depression prevention/treatment. As fish oil (FO) and Conjugated Linoleic Acid (CLA) are known Nrf2 inducer, their protective ability was comparatively evaluated in a murine model of depression (MRL/MpJ-Faslpr ). Oxidative stress, fatty acids content and critical factors reflecting brain functioning - namely brain-derived neurotrophic factor (BDNF), synaptic markers and cholinergic signaling - were preliminarily evaluated in frontal cortex of 8-weeks (Young) and in 22-weeks old animals (Old), which were used as model of depression. These markers were measured in Old mice at the end of a 5-week pre-treatment with FO or CLA (728 or 650 mg kg-1, respectively). Old mice exhibited disrupted Redox homeostasis, compensatory Nrf2 hyperactivation, lower Docosaheaxaenoic acid (DHA), lower BDNF and synaptic function proteins compared to Young mice. FO and CLA treatment relieved almost all the pathophysiological hallmarks at a level comparable to Young mice. Presented data provide the first evidence for the comparable efficacy of FO or CLA supplementation in preventing depression signs in Old MRL/lpr mice, likely through their ability of improving Nrf2-mediated antioxidant defenses.

Aluminium occurs naturally in the environment, but a variety of its compounds are produced and used in different activities of human daily life increasing so the release of this metal in the environment, particularly in water bodies. Our previous studies showed that aluminium chloride (AlCl3) induced phenotypic alterations and damages of the nervous system with decrease of the glial fibrillary acidic protein (GFAP) expression in Danio rerio fish1, optimal model organism for ecotoxicological analysis and for the study of neurodegenerative and neurobehavioral diseases2. In order to assess the toxic effects of this metal, Danio rerio embryos at shield stage were exposed to AlCl3 at the concentrations of 50, 100 and 200 μM respectively for 72 h. We compared the swimming performances of treated larvae with those of the control larvae, assessing different parameters like Distance moved, Velocity mean, Cumulative movement, Meander and Heading using the DanioVision instrument. Collected data showed that AlCl3 significantly affected the behavioural parameters with a trend inversely proportional to the concentrations, in fact the performances worsen at low concentrations compared to higher doses3. In this light, we analysed mRNA expression level by qPCR of different marker genes of neural development and function, including c-fos, appa and appb. C-fos is an immediateearly gene often used as indirect marker of neuronal activity4, while appa and appb are the homolog genes of the mammalian amyloid precursor protein (APP), an essential gene for normal brain development and a key player for the Alzheimer’s disease pathogenesis5. We observed that the expression of these genes was affected by AlCl3. The results confirmed toxic effect of AlCl3 on D. rerio larvae, suggesting the need for further experiments to uncover the mechanisms by which the aluminium exposure affects the normal developmental processes and might be at basis of neurological and behavioural disorders.

Embryos at shield stage and larvae at protruding mouth stage were exposed to different concentrations of aluminium chloride (AlCl3 ) for 72 h with the purpose to analyse their phenotype and lethality. After 24, 48 and 72 h of treatment, higher toxicity of the metal was observed on larvae with minimal lethal concentration of 0.25, 0.20 and 0.08 mm, respectively, while for embryos the corresponding values were 40, 25 and 16 mm. We observed pericardial oedema and alteration of heart rate in 50% of larvae after 48 h of exposure to 100 μm. In larvae exposed to the same concentration, there was also a neurological injury at the level of glial cells, with the number of glial fibrillary acidic protein-positive cells being significantly reduced. This study confirms the toxic nature of this metal and shows that aluminium could also interestingly represent a cardiotoxin in addition to its neurotoxic ability.

Hepatocellular carcinoma (HCC) is the third cause of cancer-related deaths worldwide. Sorafenib is the only approved drug for patients with advanced HCC but has shown limited activity. microRNAs (miRs) have been involved in several neoplasms including HCC suggesting their use or targeting as good tools for HCC treatment. The purpose of this study was to identify novel approaches to sensitize HCC cells to sorafenib through miRs. miR-423-5p was validated as positive regulator of autophagy in HCC cell lines by transient transfection of miR and anti-miR molecules. miR-423-5p expression level was evaluated by real-time polymerase chain reaction (PCR) in sera collected from 39 HCC patients before and after treatment with sorafenib. HCC cells were cotreated with sorafenib and miR-423-5p and the effects on cell cycle, apoptosis, and autophagy were evaluated. Secretory miR-423-5p was upregulated both in vitro and in vivo by sorafenib treatment and its increase was correlated with response to therapy since 75% of patients in which an increase of secretory miR423-5p was found were in partial remission or stable disease after 6 moths from the beginning of therapy. HCC cells transfected with miR-423-5p showed an increase of cell percentage in S-phase of cell cycle paralleled by a similar increase of autophagic cells evaluated at both fluorescence activated cell sorter (FACS) and transmission electron microscopy. Our results suggest the miR423-5p can be used as a useful tool to predict response to sorafenib in HCC patients and is involved in autophagy regulation in HCC cells.

Purpose: The beneficial effects of conjugated linoleic acid (CLA) mixture (cis9, trans11, c9; trans10, cis12, t10) against gliadin-induced toxicity in HLA-DQ8-transgenic mice (DQ8) have been associated with improved duodenal cytoprotective mechanisms [nuclear factor-E2-related factor- 2, Nrf2; acylpeptide hydrolase (APEH)/proteasome]. The present study was aimed at investigating the ability of individual CLA isomers to improve the efficacy of these defensive mechanisms and to protect against duodenal injury caused by the combined administration of gliadin and indomethacin (GI). Methods: Gluten-mediated enteropathy was induced in DQ8 mice by three intra-gastric administration of gliadin (20 mg kg-1/bw) and indomethacin (15 mg L-1) in drinking water for 10 days (GI). C9 or t10 CLA (520 mg kg-1/bw/day) were orally administered for 2 weeks. Pro-oxidant and toxic effects associated with GI treatment, anti-oxidant/detoxifying ability of c9 or t10-CLA and the protective effect induced by c9 pre-treatment (c9 + GI) were evaluated in DQ8 mice duodenum by combining enzymatic, immunoblotting, histological evaluation and quantitative real-time PCR assays. Results: GI treatment produces the time-dependent decline of the considered detoxifying mechanisms thus leading to pro-apoptotic and pro-oxidant effects. APEH/proteasome pathway was not markedly affected by individual CLA isomers, but duodenal redox status and activity/mRNA levels of Nrf2-activated enzymes were significantly improved by c9 administration. c9 pre-treatment protects against GI-mediated accumulation of oxidative stress markers, and histological examination reveals the increase of goblet cells number in mouse duodenum but induces only a partial recovery of APEH/proteasome activity. Conclusions: The activation of and adaptive response by low doses of c9 supplementation prevents distinct signs of gliadin-induced enteropathy in DQ8 mice

In mammals evidence that cadmium (Cd) induces apoptosis has been demonstrated in several cell-types, such as murine fibroblasts (Biagioli et al., 2001), endothelial cells (Lizard et al., 1999), renal cells (Chen & Huan, 1998) and also in the pituitary gland of rats (Poliandri et al., 2003). Less is known about the cytotoxic effects of this metal at the pituitary level in the reptiles, important bioindicators. In this study we have exposed a group of 15 lizards to an acute treatment by an intraperitoneal injection of a single and massive dose (2mg/Kg-BW) of CdCl2; another group of 15 lizards was used like control. Serial sections of 6 µm of the pituitary gland were processed for routine histological and immunohistochemical staining by ABC technique for caspase 3 and TUNEL technique for detecting apoptotic cells in situ. In the pituitary gland of P. sicula, already after 2 days the apoptotic cells appeared to increase significantly compared with those of the control. After 7 days instead, there was no considerable increment. But after 16 days the increment of apoptotic cells was much raised, above all in Rostral Pars Distalis (RPD), in which various cells showed a strongly immunostained nucleus with irregular shape. Since an increase of the apoptosis was observed above all in the RPD, we can conclude that in the lizard P. sicula Cd is particularly cytotoxic for anterior pituitary cells.