Laura Canesi
Publications
-
4.41Impact points
Immunomodulation by Different Types of N-Oxides in the Hemocytes of the Marine Bivalve Mytilus galloprovincialis.
PloS one. 01/2012; 7(5):e36937.
The potential toxicity of engineered nanoparticles (NPs) for humans and the environment represents an emerging issue. Since the aquatic environment represents the ultimate sink for NP deposition, the development of suitable assays is needed to evaluate the potential impact of NPs on aquatic biota. T... [more] The potential toxicity of engineered nanoparticles (NPs) for humans and the environment represents an emerging issue. Since the aquatic environment represents the ultimate sink for NP deposition, the development of suitable assays is needed to evaluate the potential impact of NPs on aquatic biota. The immune system is a sensitive target for NPs, and conservation of innate immunity represents an useful basis for studying common biological responses to NPs. Suspension-feeding invertebrates, such as bivalves, are particularly at risk to NP exposure, since they have extremely developed systems for uptake of nano and microscale particles integral to intracellular digestion and cellular immunity. Evaluation of the effects of NPs on functional parameters of bivalve immunocytes, the hemocytes, may help understanding the major toxic mechanisms and modes of actions that could be relevant for different NP types in aquatic organisms.In this work, a battery of assays was applied to the hemocytes of the marine bivalve Mytilus galloprovincialis to compare the in vitro effects of different n-oxides (n-TiO(2), n-SiO(2), n-ZnO, n-CeO(2)) chosen on the basis of their commercial and environmental relevance. Physico-chemical characterization of both primary particles and NP suspensions in artificial sea water-ASW was performed. Hemocyte lysosomal and mitochondrial parameters, oxyradical and nitric oxide production, phagocytic activity, as well as NP uptake, were evaluated. The results show that different n-oxides rapidly elicited differential responses hemocytes in relation to their chemical properties, concentration, behavior in sea water, and interactions with subcellular compartments. These represent the most extensive data so far available on the effects of NPs in the cells of aquatic organisms. The results indicate that Mytilus hemocytes can be utilized as a suitable model for screening the potential effects of NPs in the cells of aquatic invertebrates, and may provide a basis for future experimental work for designing environmentally safer nanomaterials.
-
3.12Impact points
-
2.86Impact points
3,5-Diiodo-L-thyronine modulates the expression of genes of lipid metabolism in a rat model of fatty liver.
The Journal of endocrinology. 11/2011; 212(2):149-58.
Recent reports demonstrated that 3,5-diiodo-l-thyronine (T(2)) was able to prevent lipid accumulation in the liver of rats fed a high-fat diet (HFD). In this study, we investigated how the rat liver responds to HFD and T(2) treatment by assessing the transcription profiles of some genes involved in ... [more] Recent reports demonstrated that 3,5-diiodo-l-thyronine (T(2)) was able to prevent lipid accumulation in the liver of rats fed a high-fat diet (HFD). In this study, we investigated how the rat liver responds to HFD and T(2) treatment by assessing the transcription profiles of some genes involved in the pathways of lipid metabolism: oxidation, storage and secretion. The mRNA levels of the peroxisome proliferator-activated receptors (PPARα, PPARγ and PPARδ), and of their target enzymes acyl-CoA oxidase and stearoyl-CoA desaturase were evaluated by real-time RT-PCR. Moreover, the expression of the adipose triglyceride lipase involved in lipid mobilisation, of the main PAT proteins acting in lipid droplet (LD) turnover, and of apoprotein B (apo B), the major protein component of very low-density lipoproteins (VLDLs) were analysed. Overall, our data demonstrated that T(2) administration to HFD rats counteracts most of the hepatic transcriptional changes that occurred in response to the excess exogenous fat. In particular, our results suggest that T(2) may prevent the pathways leading to lipid storage in LDs, promote the processes of lipid mobilisation from LDs and secretion as VLDL, in addition to the stimulation of pathways of lipid oxidation. In conclusion, our findings might give an insight into the mechanisms underlying the anti-steatotic ability of T(2) and help to define the potential therapeutic role of T(2) for preventing or treating liver steatosis.
-
2.89Impact points
Immunomodulation in Mytilus galloprovincialis by non-toxic doses of hexavalent chromium.
Fish & shellfish immunology. 09/2011; 31(6):1026-33.
In aquatic organisms, the immune function can be affected by exposure to environmental pollutants, including heavy metals. In vertebrate systems, different forms of Cr have been shown induce either immunostimulatory or immunosuppressive processes. Hexavalent Cr, Cr(VI), is an important contaminant r... [more] In aquatic organisms, the immune function can be affected by exposure to environmental pollutants, including heavy metals. In vertebrate systems, different forms of Cr have been shown induce either immunostimulatory or immunosuppressive processes. Hexavalent Cr, Cr(VI), is an important contaminant released from both domestic and industrial effluents, and the predominant chemical form of the metal in aquatic ecosystems. In this work, the in vitro and in vivo effects of Cr(VI) on immune parameters of the marine bivalve Mytilus galloprovincialis were evaluated. Hemocyte incubation with different concentrations of Cr(VI) (0.1-1-10-100 μM) induced a dose-dependent decrease in lysosomal membrane stability (LMS). Decreases in extracellular lysozyme release and phagocytic activity were also observed, with stronger effects at lower metal concentrations. On the other hand, in these conditions, Cr(VI) stimulated extracellular superoxide production and nitrite accumulation. The effects of Cr(VI) were also evaluated in mussels exposed to the metal (0.1-1-10 μg L(-1), corresponding to nanomolar concentrations) for 96 h. Decreases in hemocyte LMS values and in serum lysozyme activity were observed with increasing metal concentrations. Decreased phagocytic activity and increased NO production were recorded, with stronger effects at lower concentrations. In these conditions, decreased Total Hemocyte Counts (THC), but no necrotic/apoptotic processes were observed. Moreover, Cr(VI) at both 0.1 and 1 μg L(-1) seemed to induce significant changes in transcription of immune genes (lysozyme, Mytilin C, Myticin B, defensin, MgC1q), of the serotonin receptor (5-HTR) and of the stress protein HSP70, whereas that of the anti-apoptotic gene p53 was unaffected. Overall, the results indicate that exposure to non-toxic, environmentally relevant concentrations of Cr(VI) can modulate functional and molecular immune parameters in M. galloprovincialis.
-
3.25Impact points
Effects of dioxin exposure in Eisenia andrei: integration of biomarker data by an Expert System to rank the development of pollutant-induced stress syndrome in earthworms.
Chemosphere. 07/2011; 85(6):934-42.
A battery of biomarkers has recently been developed in the earthworm Eisenia andrei. In this study, different biomarkers (i.e. Ca²⁺-ATPase activity, lysosomal membrane stability-LMS, lysosomal lipofuscin and neutral lipid content) were utilized to evaluate the alterations in the physiological status... [more] A battery of biomarkers has recently been developed in the earthworm Eisenia andrei. In this study, different biomarkers (i.e. Ca²⁺-ATPase activity, lysosomal membrane stability-LMS, lysosomal lipofuscin and neutral lipid content) were utilized to evaluate the alterations in the physiological status of animals, induced by exposure for 3d to different sublethal concentrations of TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) (1.5 × 10⁻³, 1.5 × 10⁻², 1.5×10⁻¹ ng mL⁻¹) utilizing the paper contact toxicity test. Lysosome/cytoplasm volume ratio and DNA damage were also evaluated as a biomarker at the tissue level and as a biomarker of genotoxicity, respectively. Moreover, the NR retention time assay conditions were optimized for the determination of in vivo LMS in earthworm coelomocytes. The results demonstrate that LMS and Ca²⁺-ATPase activity were early warning biomarkers able to detect the effects of minimal amounts of TCDD and that biomarkers evaluated at the tissue level are important for following the evolution of the stress syndrome in earthworms. To evaluate the health status of the animals, an Earthworm Expert System (EES) for biomarker data integration and interpretation was developed. The EES proved to be a suitable tool able to rank, objectively, the different levels of the stress syndrome in E. andrei induced by the different concentrations of TCDD.
-
2.86Impact points
Non-receptor-mediated actions are responsible for the lipid-lowering effects of iodothyronines in FaO rat hepatoma cells.
The Journal of endocrinology. 07/2011; 210(1):59-69.
Iodothyronines influence lipid metabolism and energy homeostasis. Previous studies demonstrated that 3,5-l-diiodothyronine (T(2)), as well as 3,3',5-L-triiodothyronine (T(3)), was able to both prevent and reverse hepatic steatosis in rats fed a high-fat diet, and this effect depends on a direct ... [more] Iodothyronines influence lipid metabolism and energy homeostasis. Previous studies demonstrated that 3,5-l-diiodothyronine (T(2)), as well as 3,3',5-L-triiodothyronine (T(3)), was able to both prevent and reverse hepatic steatosis in rats fed a high-fat diet, and this effect depends on a direct action of iodothyronines on the hepatocyte. However, the involvement of thyroid hormone receptors (TRs) in mediating the lipid-lowering effect of iodothyronines was not elucidated. In this study, we investigated the ability of T(2) and T(3) to reduce the lipid overloading using the rat hepatoma FaO cells defective for functional TRs. The absence of constitutive mRNA expression of both TRα1 and TRβ1 in FaO cells was verified by RT-qPCR. To mimic the fatty liver condition, FaO cells were treated with a fatty acid mixture and then exposed to pharmacological doses of T(2) or T(3) for 24 h. Lipid accumulation, mRNA expression of the peroxisome proliferator-activated receptors (PPAR-α, -γ, -δ) the acyl-CoA oxidase (AOX), and the stearoyl CoA desaturase (SCD1), as well as fuel-stimulated O(2) consumption in intact cells, were evaluated. Lipid accumulation was associated with an increase in triacylglycerol content, PPARγ mRNA expression, and a decrease in PPARδ and SCD1 mRNA expression. The addition of T(2) or T(3) to lipid-overloaded cells resulted in i) reduction in lipid content; ii) downregulation of PPARα, PPARγ, and AOX expression; iii) increase in PPARδ expression; and iv) stimulation of mitochondrial uncoupling. These data demonstrate, for the first time, that in the hepatocyte, the lipid-lowering actions of both T(2) and T(3) are not mediated by TRs.
-
1.76Impact points
Bivalve molluscs as an unique target group for nanoparticle toxicity.
Marine environmental research. 06/2011;
Due to the continuous development and production of manufactured nanomaterials or nanoparticles (NPs), their uptake and effects in the aquatic biota represent a major concern. Estuarine and coastal environments are expected to represent the ultimate sink for NPs, where their chemical behavior (aggre... [more] Due to the continuous development and production of manufactured nanomaterials or nanoparticles (NPs), their uptake and effects in the aquatic biota represent a major concern. Estuarine and coastal environments are expected to represent the ultimate sink for NPs, where their chemical behavior (aggregation/agglomeration) and consequent fate may be critical in determining the biological impact. Bivalve mollusks are abundant from freshwater to marine ecosystems, where they are widely utilized in biomonitoring of environmental perturbations. As suspension-feeders, they have highly developed processes for cellular internalization of nano- and micro-scale particles (endo- and phagocytosis), integral to key physiological functions such as intra-cellular digestion and cellular immunity. Here we will summarise available information on the effects of different types of NPs in different bivalve species, in particular Mytilus spp. Data on the effects and modes of action of different NPs on mussel hemocytes in vitro demonstrate that cell-mediated immunity represents a significant target for NPs. Moreover, in vivo exposure to NPs indicates that, due to the physiological mechanisms involved in the feeding process, NP agglomerates/aggregates taken up by the gills are directed to the digestive gland, where intra-cellular uptake of nanosized materials induces lysosomal perturbations and oxidative stress. Overall, bivalves represent a particularly suitable model for investigating the effects and mechanisms of action underlying the potential toxicity of NPs in marine invertebrates.
-
3.25Impact points
Pleiotropic effects of hexavalent chromium (CrVI) in Mytilus galloprovincialis digestive gland.
Chemosphere. 02/2011; 83(8):1087-95.
Hexavalent Chromium Cr(VI) is an important contaminant considered as a model oxidative toxicant released from both domestic and industrial effluents, and represents the predominant chemical form of the metal in aquatic ecosystems. On the other hand, in mammals the reduced form Cr(III) is considered ... [more] Hexavalent Chromium Cr(VI) is an important contaminant considered as a model oxidative toxicant released from both domestic and industrial effluents, and represents the predominant chemical form of the metal in aquatic ecosystems. On the other hand, in mammals the reduced form Cr(III) is considered an essential microelement, involved in regulation of lipid and carbohydrate metabolism; moreover, recent evidence suggests that Cr may have endocrine effects. In this work, the effects of Cr(VI) were investigated in the digestive gland of the marine bivalve Mytilus galloprovincialis. Mussels were exposed to 0.1-1-10-100 μg Cr(VI) L(-1) animal(-1) for 96 h. At 100 μg L(-1), a large increase in total Cr tissue content was observed; in these conditions, the lysosomal membranes were completely destabilized, whereas other lysosomal biomarkers (neutral lipids-NL and lipofuscin-LF), as well as different enzyme activities and gene expression were unaffected, this indicating severe stress conditions in the tissue. On the other hand, at lower concentrations, changes in other histochemical, biochemical and molecular endpoints were observed. In particular, at both 1 and 10 μg L(-1), lysosomal destabilization was associated with significant NL and LF accumulation; however, no changes in catalase and GSH transferase (GST) activities were observed. At the same concentrations, GSSG reductase (GSR) activity was significantly increased, this probably reflecting the recycling of GSSG produced in the GSH-mediated intracellular reduction of Cr(VI). Increased activities of the key glycolytic enzymes PFK (phosphofructokinase) and PK (pyruvate kinase) were also observed, indicating that Cr(VI) could affect carbohydrate metabolism. Cr(VI) induced downregulation or no effects on the expression of metallothioneins MT10 and MT20, except for an increase in MT20 transcription in males. Moreover, significant up-regulation of the Mytilus estrogen receptor MeER2 and serotonin receptor (5-HTR) were observed in both sexes. The results demonstrate that exposure to Cr(VI) in the low ppb range did not result in strong toxicity or oxidative stress conditions in mussel digestive gland. On the other hand, our data support the hypothesis that low concentrations of the metal can exert pleiotropic effects on mussel physiology, from modulation of lipid and carbohydrate metabolism, to effects on the expression of estrogen-responsive genes.
-
1.75Impact points
Effects of fruit and vegetable low molecular mass fractions on gene expression in gingival cells challenged with Prevotella intermedia and Actinomyces naeslundii.
Journal of biomedicine & biotechnology. 01/2011; 2011:230630.
Low molecular mass (LMM) fractions obtained from extracts of raspberry, red chicory, and Shiitake mushrooms have been shown to be an useful source of specific antibacterial, antiadhesion/coaggregation, and antibiofilm agent(s) that might be used for protection towards caries and gingivitis. In this ... [more] Low molecular mass (LMM) fractions obtained from extracts of raspberry, red chicory, and Shiitake mushrooms have been shown to be an useful source of specific antibacterial, antiadhesion/coaggregation, and antibiofilm agent(s) that might be used for protection towards caries and gingivitis. In this paper, the effects of such LMM fractions on human gingival KB cells exposed to the periodontal pathogens Prevotella intermedia and Actinomyces naeslundii were evaluated. Expression of cytokeratin 18 (CK18) and β4 integrin (β4INT) genes, that are involved in cell proliferation/differentiation and adhesion, and of the antimicrobial peptide β2 defensin (HβD2) in KB cells was increased upon exposure to either live or heat-killed bacteria. All LMM fractions tested prevented or reduced the induction of gene expression by P. intermedia and A. naeslundii depending on the experimental conditions. Overall, the results suggested that LMM fractions could modulate the effects of bacteria associated with periodontal disease in gingival cells.
-
4.41Impact points
The organophosphate Chlorpyrifos interferes with the responses to 17β-estradiol in the digestive gland of the marine mussel Mytilus galloprovincialis.
PloS one. 01/2011; 6(5):e19803.
Many pesticides have been shown to act as endocrine disrupters. Although the potencies of currently used pesticides as hormone agonists/antagonists are low compared with those of natural ligands, their ability to act via multiple mechanisms might enhance the biological effect. The organophosphate Ch... [more] Many pesticides have been shown to act as endocrine disrupters. Although the potencies of currently used pesticides as hormone agonists/antagonists are low compared with those of natural ligands, their ability to act via multiple mechanisms might enhance the biological effect. The organophosphate Chlorpyrifos (CHP) has been shown to be weakly estrogenic and cause adverse neurodevelopmental effects in mammals. However, no information is available on the endocrine effects of CHP in aquatic organisms. In the digestive gland of the bivalve Mytilus galloprovincialis, a target tissue of both estrogens and pesticides, the possible effects of CHP on the responses to the natural estrogen 17β-estradiol (E(2)) were investigated. Mussels were exposed to CHP (4.5 mg/l, 72 hrs) and subsequently injected with E(2) (6.75 ng/g dw). Responses were evaluated in CHP, E(2) and CHP/E(2) treatment groups at 24 h p.i. by a biomarker/transcriptomic approach. CHP and E(2) induced additive, synergistic, and antagonistic effects on lysosomal biomarkers (lysosomal membrane stability, lysosome/cytoplasm volume ratio, lipofuscin and neutral lipid accumulation). Additive and synergistic effects were also observed on the expression of estrogen-responsive genes (GSTπ, catalase, 5-HTR) evaluated by RT-Q-PCR. The use of a 1.7K cDNA Mytilus microarray showed that CHP, E(2) and CHP/E(2), induced 81, 44, and 65 Differentially Expressed Genes (DEGs), respectively. 24 genes were exclusively shared between CHP and CHP/E(2), only 2 genes between E(2) and CHP/E(2). Moreover, 36 genes were uniquely modulated by CHP/E(2). Gene ontology annotation was used to elucidate the putative mechanisms involved in the responses elicited by different treatments. The results show complex interactions between CHP and E(2) in the digestive gland, indicating that the combination of certain pesticides and hormones may give rise to unexpected effects at the molecular/cellular level. Overall, these data demonstrate that CHP can interfere with the mussel responses to natural estrogens.
-
2.27Impact points
Metals, metallothioneins and oxidative stress in blood of autistic children
Research in Autism Spectrum Disorders. 01/2011; 5:286-293.
Many factors have been implicated in autism onset, including excess or deficiency in toxic or essential metals and impaired antioxidant systems. Protection towards the damaging effects of reactive oxygen species (ROS) is afforded by antioxidant enzymes (superoxide dismutase, SOD, catalase, CAT, glut... [more] Many factors have been implicated in autism onset, including excess or deficiency in toxic or essential metals and impaired antioxidant systems. Protection towards the damaging effects of reactive oxygen species (ROS) is afforded by antioxidant enzymes (superoxide dismutase, SOD, catalase, CAT, glutathione peroxidase, GPx), and non-enzymatic antioxidants such as metallothionein (MT). In blood samples from autistic children and healthy controls, three aspects were investigated: (i) the plasma concentration of 13 metals (Al, As, Ca, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Si, Zn) was measured by ICP-AES; (ii) the transcription of three MT isoforms (MT-1A, MT-1E and MT-2A) was assessed by real-time RT-PCR in peripheral blood leucocytes (PBMCs); (iii) the activity of SOD, CAT and GPx was assessed by spectrophotometric assays in red blood cells (RBCs). Autistic children showed significantly higher plasma levels of Zn, Ca, Fe, As, Ni, Cd and Si, higher mRNA expression of the MT isoforms in PBMCs, higher SOD activity and lower CAT activity in RBCs with respect to controls. These findings demonstrate a significant increase in both metal content and metallothionein expression together with an unbalance in the oxidative status in the blood of autistic children.
-
7.82Impact points
Direct effects of iodothyronines on excess fat storage in rat hepatocytes.
Journal of hepatology. 11/2010; 54(6):1230-6.
Previous studies have demonstrated that 3,5-L-diiodothyronine (T(2)) is able to prevent lipid accumulation in the liver of rats fed a high-fat diet. Whether this effect is due to a direct action of T(2) on the liver has not been elucidated. In this study, we investigated the ability of T(2) to reduc... [more] Previous studies have demonstrated that 3,5-L-diiodothyronine (T(2)) is able to prevent lipid accumulation in the liver of rats fed a high-fat diet. Whether this effect is due to a direct action of T(2) on the liver has not been elucidated. In this study, we investigated the ability of T(2) to reduce the excess lipids in isolated hepatocytes treated with fatty acids (FFAs). The effects of T(2) were compared with those elicited by 3,3',5-L-triiodothyronine (T(3)). To mimic the fatty liver condition, primary cultures of rat hepatocytes were overloaded with lipids, by exposure to FFAs ("fatty hepatocytes"), and then treated with T(2) or T(3). Lipid content, morphometry of lipid droplets (LDs), and expression of the adipocyte differentiation-related protein (ADRP) and the peroxisome proliferator-activated receptors (PPAR-α, -γ, -δ) were evaluated. Activities of the lipolytic enzyme acyl CoA oxidase-AOX and the antioxidant enzymes superoxide dismutase-SOD and catalase-CAT were also determined. FFA-induced lipid accumulation was associated with an increase in both number/size of LDs and expression of ADRP, PPAR-γ, and PPAR-δ/β mRNAs, as well as in the activities of AOX, SOD, and CAT. The addition of T(2) or T(3) to "fatty hepatocytes" resulted in a reduction in: (i) lipid content and LD diameter; (ii) PPAR-γ and PPAR-δ expression; (iii) activities of AOX and antioxidant enzymes. These data demonstrate, for the first time, a direct action of both T(2) and T(3) in reducing the excess fat in cultured hepatocytes.
-
6.40Impact points
Melatonin signaling and cell protection function.
FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 10/2010; 24(10):3603-24.
Besides its well-known regulatory role on circadian rhythm, the pineal gland hormone melatonin has other biological functions and a distinct metabolism in various cell types and peripheral tissues. In different tissues and organs, melatonin has been described to act as a paracrine and also as an int... [more] Besides its well-known regulatory role on circadian rhythm, the pineal gland hormone melatonin has other biological functions and a distinct metabolism in various cell types and peripheral tissues. In different tissues and organs, melatonin has been described to act as a paracrine and also as an intracrine and autocrine agent with overall homeostatic functions and pleiotropic effects that include cell protection and prosurvival factor. These latter effects, documented in a number of in vitro and in vivo studies, are sustained through both receptor-dependent and -independent mechanisms that control detoxification and stress response genes, thus conferring protection against a number of xenobiotics and endobiotics produced by acute and chronic noxious stimuli. Redox-sensitive components are included in the cell protection signaling of melatonin and in the resulting transcriptional response that involves the control of NF-κB, AP-1, and Nrf2. By these pathways, melatonin stimulates the expression of antioxidant and detoxification genes, acting in turn as a glutathione system enhancer. A further and converging mechanism of cell protection by this indoleamine described in different models seems to lie in the control of damage and signaling function of mitochondria that involves decreased production of reactive oxygen species and activation of the antiapoptotic and redox-sensitive element Bcl2. Recent evidence suggests that upstream components in this mitochondrial route include the calmodulin pathway with its central role in melatonin signaling and the survival-promoting component of MAPKs, ERK1/2. In this review article, we will discuss these and other molecular aspects of melatonin signaling relevant to cell protection and survival mechanisms.
-
1.81Impact points
Specificity of anti-Vibrio immune response through p38 MAPK and PKC activation in the hemocytes of the mussel Mytilus galloprovincialis.
Journal of invertebrate pathology. 09/2010; 105(1):49-55.
In mussel (Mytilus sp.) hemocytes, differential functional responses to injection with different types of live and heat-killed Vibrio species have been recently demonstrated. In this work, responses of Mytilus hemocytes to heat-killed Vibrio splendidus LGP32 and the mechanisms involved were investig... [more] In mussel (Mytilus sp.) hemocytes, differential functional responses to injection with different types of live and heat-killed Vibrio species have been recently demonstrated. In this work, responses of Mytilus hemocytes to heat-killed Vibrio splendidus LGP32 and the mechanisms involved were investigated in vitro and the results were compared with those obtained with Vibrio anguillarum (ATCC 19264). Adhesion of hemocytes after incubation with bacteria was evaluated by flow cytometry: both total hemocyte counts (THC) and percentage of hemocyte sub-populations were determined in non-adherent cells. Functional parameters such as lysosomal membrane stability, lysozyme release, extracellular ROS production and NO production were evaluated, as well as the phosphorylation state of the stress-activated p38 MAPK and PKC. Neither Vibrio affected total hemocyte adhesion, while both induced similar lysosomal destabilization and NO production. However, V. splendidus decreased adhesion of large granulocytes, induced rapid and persistent lysozyme release and stimulated extracellular ROS production: these effects were associated with persistent activation of p38 MAPK and PKC. In contrast, V. anguillarum decreased adhesion of large semigranular hemocytes and increased that of hyalinocytes, had no effect on the extracellular ROS production, and induced significantly lower lysozyme release and phosphorylation of p-38 MAPK and PKC than V. splendidus. These data reinforced the existence of specific interactions between mussel hemocytes and V. splendidus LGP32 and suggest that this Vibrio strain affects bivalve hemocytes through disregulation of immune signaling. The results support the hypothesis that responses of bivalve hemocytes to different bacterial stimuli may depend not only on the nature of the stimulus, but also on the cell subtype, thus leading to differential activation of signaling components.
-
2.58Impact points
Effects of vibrio challenge on digestive gland biomarkers and antioxidant gene expression in Mytilus galloprovincialis.
Comparative biochemistry and physiology. Toxicology & pharmacology : CBP. 09/2010; 152(3):399-406.
In bivalve molluscs, responses to bacterial infection have been largely characterized in terms of both functional responses and gene expression in the immune cells, the hemocytes. The effects of bacterial challenge at the tissue level, where bacterial infection may cause stressful conditions, have n... [more] In bivalve molluscs, responses to bacterial infection have been largely characterized in terms of both functional responses and gene expression in the immune cells, the hemocytes. The effects of bacterial challenge at the tissue level, where bacterial infection may cause stressful conditions, have not been so far specifically investigated. Biomarkers are widely utilised to evaluate the health status of bivalves, from the molecular to the organism level, in response to both natural and anthropogenic stressors. In this work, the effects of in vivo challenge with heat-killed vibrio species, Vibrio splendidus LGP32 and Vibrio anguillarum (ATCC19264), on different biomarkers in the digestive gland of the marine bivalve Mytilus galloprovincialis were investigated. Mussels were injected with either vibrio and tissues sampled at 3, 6 and 24 h post injection (p.i.). Lysosomal biomarkers, such as lysosomal membrane stability (LMS) and lipofuscin accumulation, as well as specific activities of antioxidant enzymes (catalase and glutathione transferase-GST) were evaluated. Moreover, the expression of antioxidant molecules (catalase, GST-pi and metallothioneins MT10 and MT20) was determined by quantitative RT-PCR. Both V. splendidus and V. anguillarum significantly affected all parameters measured, to a different extent and at different times p.i. Interestingly, whereas both vibrios induced lysosomal membrane destabilisation and increases in the activities of antioxidant enzymes, distinct responses were observed in terms of lysosomal lipofuscin accumulation and expression of antioxidant molecules. In particular, V. splendidus induced a general increase in the transcription of antioxidant genes, indicating that Mytilus digestive gland can mount an efficient antioxidant response towards this vibrio species. On the other hand, a general down-regulation or no effect was observed with V. anguillarum. The lack of this response was reflected in stronger oxidative stress conditions in the digestive gland of mussels challenged with V. anguillarum, as indicated by higher levels of lysosomal lipofuscin observed at longer times p.i. Overall, these data indicate that lysosomal and oxidative stress biomarkers could be usefully applied in order to monitor early changes in the health status of bivalves induced by bacteria. Moreover, the results support the hypothesis that host responses to bacteria may be taken into account when interpreting biomarker data in ecotoxicological studies.
-
2.73Impact points
Aromatase mRNA expression in the brain of adult Xenopus laevis exposed to Lambro river water and endocrine disrupting compounds.
General and comparative endocrinology. 09/2010; 168(2):262-8.
Aromatase P450 (P450 arom; Cyp19) is a key enzyme for vertebrate reproduction and brain development that catalyzes the conversion of androgens to estrogens. The aim of this study was to improve the knowledge on EDC effects by analysing their potential impact on brain P450 arom in adult Xenopus laevi... [more] Aromatase P450 (P450 arom; Cyp19) is a key enzyme for vertebrate reproduction and brain development that catalyzes the conversion of androgens to estrogens. The aim of this study was to improve the knowledge on EDC effects by analysing their potential impact on brain P450 arom in adult Xenopus laevis exposed for 4 weeks to an environmental sample, the water of the river Lambro (LAM), the most polluted tributary of the Po river in North Italy. Other groups were exposed to individual compounds 10(-8) M tamoxifen (TAM), ethinylestradiol (EE2), flutamide (FLU) and methyldihydrotestosterone (MDHT) known for their (anti)estrogenic and (anti)androgenic modes of action. Expression of CYP19 was evaluated in brain extracts by quantitative RT-PCR, using a pair of primers located in the open reading frame (ORF) that allowed the simultaneous amplification of all transcripts (Aro-ORF) and a pair of primers specific for brain aromatase (Aro-B). Significant increase in Aro-ORF and Aro-B mRNA levels were observed in both females and males exposed to LAM. Different changes were observed for the model compounds using two pairs of primers. Aro-ORF mRNA expression was significantly increased in EE2 and MDHT exposed males and in FLU-exposed females, while it was significantly decreased in TAM exposed females. Aro-B mRNA was significantly increased in both sexes exposed to FLU and decreased in TAM exposed females. In conclusion, aromatase mRNA in the brain of X. laevis was regulated differentially in a gender specific manner by certain (anti)estrogenic and (anti)androgenic EDCs, supporting previous hypotheses that diverse compounds present in the river Lambro may induce feminization and demasculinization effects.
-
1.98Impact points
PAT protein mRNA expression in primary rat hepatocytes: Effects of exposure to fatty acids.
International journal of molecular medicine. 04/2010; 25(4):505-12.
Excess energy is stored as neutral lipids in lipid droplets (LDs) whose surface is coated by PAT proteins, each playing a distinct cellular function. The adipocyte differentiation-related protein (ADRP) and tail-interacting protein (TIP47) are expressed almost ubiquitously, whereas the oxidative tis... [more] Excess energy is stored as neutral lipids in lipid droplets (LDs) whose surface is coated by PAT proteins, each playing a distinct cellular function. The adipocyte differentiation-related protein (ADRP) and tail-interacting protein (TIP47) are expressed almost ubiquitously, whereas the oxidative tissue-enriched PAT protein (OXPAT) is expressed in specific tissues, such as the liver. In rat liver, only ADRP expression has been documented. This study was aimed at identifying OXPAT and TIP47 transcripts in rat hepatocytes, and investigating how their expression is modulated by excess lipids, using fat-enriched hepatocytes to mimic different degrees of steatosis. Primary rat hepatocytes were exposed to fatty acids (FFAs) for 12, 24 and 36 h. Lipid accumulation was estimated by spectrophotometric quantification of triacylglycerol. Expression of PAT proteins as well as of PPARgamma was evaluated by real-time RT-PCR. Hepatocytes exposed to FFAs showed progressive lipid accumulation. The increase in lipid content was associated with the induction of PAT protein expression. At 12 h, OXPAT and TIP47 mRNA expression was up-regulated. At longer times, the level of OXPAT transcripts remained high, whereas that of TIP47 slowly declined. Conversely, ADRP expression showed a time-dependent increase with exposure to FFAs. This study demonstrates, for the first time, the presence of OXPAT and TIP47 transcripts in rat hepatocytes, as well as their up-regulation with lipid accumulation. The distinct time courses observed for the three PAT proteins during FFA exposure might reflect the different roles played by each protein in lipid metabolism in the hepatocyte. Up-regulation of TIP47 and OXPAT might represent an early response to excess lipids, while, in correspondence with a lipid overload, up-regulation of ADRP could address lipids towards storage.
-
3.12Impact points
Biomarkers in Mytilus galloprovincialis exposed to suspensions of selected nanoparticles (Nano carbon black, C60 fullerene, Nano-TiO2, Nano-SiO2).
Aquatic toxicology (Amsterdam, Netherlands). 04/2010; 100(2):168-77.
The potential for ecological toxicity associated with nanomaterials is a growing area of investigation, in particular in the aquatic environment. In suspension feeding invertebrates, the cellular immune system and digestive gland are likely to be targeted, due to their highly developed processes for... [more] The potential for ecological toxicity associated with nanomaterials is a growing area of investigation, in particular in the aquatic environment. In suspension feeding invertebrates, the cellular immune system and digestive gland are likely to be targeted, due to their highly developed processes for the cellular internalisation of nano- and micro-scale particles that are integral to key physiological functions such as cellular immunity and intracellular digestion. We have recently demonstrated that suspensions of selected commercial nanomaterials, namely Nano carbon black (NCB), C60 fullerene (C60), Nano-titanium dioxide (n-TiO2) and Nanosilica (n-SiO2) induce oxyradical production and lysosomal enzyme release in the hemocytes of the marine mussel Mytilus in vitro. In this work the possible effects of in vivo exposure to these NPs were investigated. Mussels were exposed to different concentrations (0.05-0.2-1-5mg/l) of NP suspensions for 24h and different biomarkers were evaluated in hemocytes, digestive gland and gills. Characterisation of NP suspensions in artificial sea water (ASW) was performed, indicating the formation of agglomerates of different sizes in the nano-micromolar range for different types of NPs. Formation of larger agglomerates was observed at the end of exposure. The results show that all NP suspensions induced significant lysosomal membrane destabilisation in both the hemocytes and the digestive gland, with NCB>C60>n-TiO2, >n-SiO2. In the digestive gland, all NPs induced lysosomal lipofuscin accumulation only at the highest concentrations tested to a different extent depending on the NP type. NCB, TiO2 and SiO2 also induced lysosomal neutral lipid accumulation. Moreover, all NPs increased the activity of the antioxidant enzyme catalase, with n-SiO2>NCB≅TiO2>C60; NCB and n-TiO2 also stimulated glutathione transferase (GST). Changes in catalase and GST activities were also observed in gills, with both increases and decreases depending on NP type and concentration. The reported results demonstrate that in mussels responses to exposure to NP suspensions involve changes in lysosomal and oxidative stress biomarkers in the digestive gland, suggesting uptake of NP aggregates/agglomerates mainly through the digestive system. Overall, these data further support the hypothesis that suspension feeding invertebrates represent a significant target for NPs in the aquatic environment.
-
3.12Impact points
In vitro effects of suspensions of selected nanoparticles (C60 fullerene, TiO(2), SiO(2)) on Mytilus hemocytes.
Aquatic toxicology (Amsterdam, Netherlands). 10/2009;
As the nanotechnology industries increase production, nanoscale products will enter the aquatic environment, posing a possible threat to aquatic organisms. Suspension-feeding invertebrates may represent a unique target group for nanoparticle (NP) ecotoxicity, since they have highly developed process... [more] As the nanotechnology industries increase production, nanoscale products will enter the aquatic environment, posing a possible threat to aquatic organisms. Suspension-feeding invertebrates may represent a unique target group for nanoparticle (NP) ecotoxicity, since they have highly developed processes for the cellular internalisation of nano- and microscale particles (endocytosis and phagocytosis), which are integral to key physiological functions such as intracellular digestion and cellular immunity. In the marine bivalve Mytilus, short-term exposure to nanosized carbon black (NCB) was shown to significantly affect immune parameters of immune cells, the hemocytes, in vitro. In this work, we further investigated the effects of other types of commercial NPs (C60 fullerene, TiO(2) and SiO(2) at 1, 5, 10mug/ml) on Mytilus hemocytes. Characterization of NP suspensions in artificial sea water (ASW) was performed, indicating the formation of agglomerates of different sizes for different types of NPs. None of the NP tested significantly affected lysosomal membrane stability, indicating the lack of a major toxic effect. However, all NP suspensions induced a concentration-dependent lysozyme release, extracellular oxyradical and nitric oxide (NO) production, to a different extent and with different time courses depending on the concentration and the NP type. The inflammatory effects of NPs were mediated by rapid activation of the stress-activated p38 MAPK. The results further support the hypothesis that in bivalves the immune system represents a significant target for NPs.
-
4.00Impact points
Neuroprotective mesenchymal stem cells are endowed with a potent antioxidant effect in vivo.
Journal of neurochemistry. 08/2009;
Experimental autoimmune encephalomyelitis (EAE), an animal model for human multiple sclerosis, is characterized by demyelination, inflammation and neurodegeneration of CNS in which free radicals play a role. Recently, the efficacy of murine mesenchimal stem cells (MSCs) as treatment of EAE induced i... [more] Experimental autoimmune encephalomyelitis (EAE), an animal model for human multiple sclerosis, is characterized by demyelination, inflammation and neurodegeneration of CNS in which free radicals play a role. Recently, the efficacy of murine mesenchimal stem cells (MSCs) as treatment of EAE induced in mice by the encephalitogenic peptide MOG(35-55) was demonstrated. The present study analyzed some markers of oxidative stress, inflammation/degeneration and apoptosis such as metallothioneins (MTs), antioxidant enzymes (superoxide dismutase, catalase and glutathione-S-transferase), poly(ADP-ribose) polymerase-1 and p53 during EAE progression and following MSC treatment. Expression of the three brain MT isoforms increased significantly in EAE mice compared with healthy controls, but while expression of MT-1 and MT-3 increased along EAE course, MT-2 was up-regulated at the onset, but returned to levels similar to those of controls in chronic phase. The changes in the transcription and activity of the antioxidant enzymes and in expression of poly(ADP-ribose) polymerase-1 and p53 showed the same kinetics observed for MT-1 and MT-3 during EAE. Interestingly, i.v. administration of MSCs reduced the EAE-induced increases in levels/activities of all these proteins. These results support an antioxidant and neuroprotective activity for MSCs that was also confirmed in vitro on neuroblastoma cells exposed to an oxidative insult.
Following (13)
-
Isabella Panfoli
Università degli Studi di Genova -
Alessandro Dagnino
University of Eastern Piedmont -
Elena Grasselli
Università degli Studi di Genova -
Elia Ranzato
Università degli Studi del Piemonte Orientale "Amedeo Avogadro" -
Antonia Lanni
Second University of Naples
91
Publications
15
Followers
