197 results on '"Carlo Storelli"'
Search Results
2. Comparison among Different Gilthead Sea Bream (Sparus aurata) Farming Systems: Activity of Intestinal and Hepatic Enzymes and 13C-NMR Analysis of Lipids
- Author
-
Vincenzo Zonno, Francesco Paolo Fanizzi, Carlo Storelli, Giorgia Bressani, Sandra A. De Pascali, Laura Del Coco, and Paride Papadia
- Subjects
gilthead sea bream (Sparus aurata) ,PUFA ,rearing systems ,food authenticity ,nutritional physiological state ,13C NMR profiling ,Nutrition. Foods and food supply ,TX341-641 - Abstract
In order to evaluate differences in general health and nutritional values of gilthead sea bream (Sparus aurata), the effects of semi-intensive, land-based tanks and sea-cages intensive rearing systems were investigated, and results compared with captured wild fish. The physiological state was determined by measuring the activity of three different intestinal digestive enzymes: alkaline phosphatase (ALP), leucine aminopeptidase (LAP) and maltase; and the activity of the hepatic ALP. Also, the hepatic content in protein, cholesterol, and lipid were assessed. 13C-NMR analysis for qualitative and quantitative characterization of the lipid fraction extracted from fish muscles for semiintensive and land based tanks intensive systems was performed. The lipid fraction composition showed small but significant differences in the monounsaturated/saturated fatty acid ratio, with the semi-intensive characterized by higher monounsaturated and lower saturated fatty acid content with respect to land based tanks intensive rearing system.
- Published
- 2009
- Full Text
- View/download PDF
3. Additive regulation of adiponectin expression by the mediterranean diet olive oil components oleic Acid and hydroxytyrosol in human adipocytes.
- Author
-
Egeria Scoditti, Marika Massaro, Maria Annunziata Carluccio, Mariangela Pellegrino, Martin Wabitsch, Nadia Calabriso, Carlo Storelli, and Raffaele De Caterina
- Subjects
Medicine ,Science - Abstract
Adiponectin, an adipocyte-derived insulin-sensitizing and anti-inflammatory hormone, is suppressed in obesity through mechanisms involving chronic inflammation and oxidative stress. Olive oil consumption is associated with beneficial cardiometabolic actions, with possible contributions from the antioxidant phenol hydroxytyrosol (HT) and the monounsaturated fatty acid oleic acid (OA, 18:1n-9 cis), both possessing anti-inflammatory and vasculo-protective properties. We determined the effects of HT and OA, alone and in combination, on adiponectin expression in human and murine adipocytes under pro-inflammatory conditions induced by the cytokine tumor necrosis factor(TNF)-α. We used human Simpson-Golabi-Behmel syndrome (SGBS) adipocytes and murine 3T3-L1 adipocytes as cell model systems, and pretreated them with 1-100 μmol/L OA, 0.1-20 μmol/L HT or OA plus HT combination before stimulation with 10 ng/mL TNF-α. OA or HT significantly (P
- Published
- 2015
- Full Text
- View/download PDF
4. Transcriptome-based identification of new anti-inflammatory and vasodilating properties of the n-3 fatty acid docosahexaenoic acid in vascular endothelial cell under proinflammatory conditions [corrected].
- Author
-
Marika Massaro, Rosanna Martinelli, Valentina Gatta, Egeria Scoditti, Mariangela Pellegrino, Maria Annunziata Carluccio, Nadia Calabriso, Tonia Buonomo, Liborio Stuppia, Carlo Storelli, and Raffaele De Caterina
- Subjects
Medicine ,Science - Abstract
High intakes of n-3 fatty acids exert anti-inflammatory effects and cardiovascular protection, but the underlying molecular basis is incompletely defined. By genome-wide analysis we searched for novel effects of docosahexaenoic acid (DHA) on gene expression and pathways in human vascular endothelium under pro-inflammatory conditions.Human umbilical vein endothelial cells were treated with DHA and then stimulated with interleukin(IL)-1β. Total RNA was extracted, and gene expression examined by DNA microarray. DHA alone altered the expression of 188 genes, decreasing 92 and increasing 96. IL-1β changed the expression of 2031 genes, decreasing 997 and increasing 1034. Treatment with DHA before stimulation significantly affected the expression of 116 IL-1β-deregulated genes, counter-regulating the expression of 55 genes among those decreased and of 61 among those increased. Functional and network analyses identified immunological, inflammatory and metabolic pathways as the most affected. Newly identified DHA-regulated genes are involved in stemness, cellular growth, cardiovascular system function and cancer, and included cytochrome p450 4F2(CYP4F2), transforming growth factor(TGF)-β2, Cluster of Differentiation (CD)47, caspase recruitment domain(CARD)11 and phosphodiesterase(PDE)5α.Endothelial exposure to DHA regulates novel genes and related pathways. Such unbiased identification should increase our understanding of mechanisms by which n-3 fatty acids affect human diseases.
- Published
- 2015
- Full Text
- View/download PDF
5. Anti-aggregating effect of the naturally occurring dipeptide carnosine on aβ1-42 fibril formation.
- Author
-
Alessandra Aloisi, Amilcare Barca, Alessandro Romano, Sara Guerrieri, Carlo Storelli, Rosaria Rinaldi, and Tiziano Verri
- Subjects
Medicine ,Science - Abstract
Carnosine is an endogenous dipeptide abundant in the central nervous system, where by acting as intracellular pH buffering molecule, Zn/Cu ion chelator, antioxidant and anti-crosslinking agent, it exerts a well-recognized multi-protective homeostatic function for neuronal and non-neuronal cells. Carnosine seems to counteract proteotoxicity and protein accumulation in neurodegenerative conditions, such as Alzheimer's Disease (AD). However, its direct impact on the dynamics of AD-related fibril formation remains uninvestigated. We considered the effects of carnosine on the formation of fibrils/aggregates of the amyloidogenic peptide fragment Aβ1-42, a major hallmark of AD injury. Atomic force microscopy and thioflavin T assays showed inhibition of Aβ1-42 fibrillogenesis in vitro and differences in the aggregation state of Aβ1-42 small pre-fibrillar structures (monomers and small oligomers) in the presence of carnosine. in silico molecular docking supported the experimental data, calculating possible conformational carnosine/Aβ1-42 interactions. Overall, our results suggest an effective role of carnosine against Aβ1-42 aggregation.
- Published
- 2013
- Full Text
- View/download PDF
6. Di- and tripeptide transport in vertebrates: the contribution of teleost fish models
- Author
-
Amilcare Barca, Barbara Piccinni, Tiziano Verri, Alessandro Romano, Paola Pisani, Carlo Storelli, Verri, Tiziano, Barca, Amilcare, Pisani, Paola, Piccinni, Barbara, Storelli, Carlo, and Romano, Alessandro
- Subjects
0301 basic medicine ,Fish Proteins ,Dietary protein ,Physiology ,Protein digestion ,Tripeptide ,Growth ,Biology ,Biochemistry ,Peptide transport ,Intestinal absorption ,Digestive physiology ,03 medical and health sciences ,Endocrinology ,Osmoregulation ,Animals ,Humans ,Compensatory growth ,Adaptation ,Epithelial physiology ,Ecology, Evolution, Behavior and Systematics ,Symporters ,Circadian rhythm ,Feeding ,Tripeptide transport ,Fishes ,Immune system physiology ,Fasting ,Food deprivation ,Solute carrier family ,Renal physiology ,030104 developmental biology ,Models, Animal ,Ontogeny ,Peptide absorption ,Animal Science and Zoology ,Cotransporter ,Oligopeptides - Abstract
Solute Carrier 15 (SLC15) family, alias H(+)-coupled oligopeptide cotransporter family, is a group of membrane transporters known for their role in the cellular uptake of di- and tripeptides (di/tripeptides) and peptide-like molecules. Of its members, SLC15A1 (PEPT1) chiefly mediates intestinal absorption of luminal di/tripeptides from dietary protein digestion, while SLC15A2 (PEPT2) mainly allows renal tubular reabsorption of di/tripeptides from ultrafiltration, SLC15A3 (PHT2) and SLC15A4 (PHT1) possibly interact with di/tripeptides and histidine in certain immune cells, and SLC15A5 has unknown function. Our understanding of this family in vertebrates has steadily increased, also due to the surge of genomic-to-functional information from 'non-conventional' animal models, livestock, poultry, and aquaculture fish species. Here, we review the literature on the SLC15 transporters in teleost fish with emphasis on SLC15A1 (PEPT1), one of the solute carriers better studied amongst teleost fish because of its relevance in animal nutrition. We report on the operativity of the transporter, the molecular diversity, and multiplicity of structural-functional solutions of the teleost fish orthologs with respect to higher vertebrates, its relevance at the intersection of the alimentary and osmoregulative functions of the gut, its response under various physiological states and dietary solicitations, and its possible involvement in examples of total body plasticity, such as growth and compensatory growth. By a comparative approach, we also review the few studies in teleost fish on SLC15A2 (PEPT2), SLC15A4 (PHT1), and SLC15A3 (PHT2). By representing the contribution of teleost fish to the knowledge of the physiology of di/tripeptide transport and transporters, we aim to fill the gap between higher and lower vertebrates.
- Published
- 2017
7. Hydroxytyrosol suppresses MMP-9 and COX-2 activity and expression in activated human monocytes via PKCα and PKCβ1 inhibition
- Author
-
Carlo Storelli, Maria Annunziata Carluccio, Nadia Calabriso, Marika Massaro, Alessia Nestola, Raffaele De Caterina, Egeria Scoditti, Egeria, Scoditti, Alessia, Nestola, Marika, Massaro, Nadia, Calabriso, Storelli, Carlo, Raffaele De, Caterina, and Maria Annunziata, Carluccio
- Subjects
Protein Kinase C-alpha ,Down-Regulation ,Inflammation ,Matrix metalloproteinase ,Pharmacology ,Diet, Mediterranean ,Peripheral blood mononuclear cell ,Gene Expression Regulation, Enzymologic ,Monocytes ,chemistry.chemical_compound ,Phorbol Esters ,Protein Kinase C beta ,medicine ,Humans ,Plant Oils ,Zymography ,Olive Oil ,Protein kinase C ,biology ,Macrophages ,Monocyte ,NF-kappa B p50 Subunit ,Polyphenols ,U937 Cells ,Phenylethyl Alcohol ,Enzyme Activation ,Oxidative Stress ,medicine.anatomical_structure ,Matrix Metalloproteinase 9 ,chemistry ,Biochemistry ,Cyclooxygenase 2 ,Leukocytes, Mononuclear ,biology.protein ,Hydroxytyrosol ,Cyclooxygenase ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,Oxidation-Reduction ,Signal Transduction - Abstract
Hydroxytyrosol (HT), the major olive oil antioxidant polyphenol in cardioprotective Mediterranean diets, is endowed with anti-inflammatory and anti-atherosclerotic activity. The production of cyclooxygenase (COX)-2-dependent inflammatory eicosanoids and the functionally linked release of matrix metalloproteinase (MMP)-9 by macrophages likely contribute to plaque instability leading to acute coronary events. Objective of the study was to examine the HT effects on inflammatory markers in human activated monocytes, including MMP-9 and COX-2 activity and expression and explore HT underlying mechanisms.Human peripheral blood mononuclear cells (PBMC) and U937 monocytes were treated with 1-10 μmol/L HT before activation with phorbol myristate acetate (PMA). HT blunted monocyte matrix invasive potential and reduced MMP-9 release and expression at zymography, ELISA and RT-PCR, with an IC50 = 10 μmol/L ( P0.05), without affecting tissue inhibitor of metalloproteinase (TIMP)-1. Moreover, HT inhibited prostaglandin (PG)E2 production and COX-2 expression, without affecting COX-1. These effects were mediated by inhibition of transcription factor nuclear factor (NF)-κB and protein kinase C (PKC)α and PKCβ1 activation.HT, at nutritionally relevant concentrations, reduces MMP-9 and COX-2 induction in activated human monocytes via PKCα and PKCβ1 inhibition, thus featuring novel anti-inflammatory properties. Overall, such results contribute to explaining the vascular protective effects by olive oil polyphenols in Mediterranean diets.
- Published
- 2014
8. Teleost fish models in membrane transport research: the PEPT1(SLC15A1) H+-oligopeptide transporter as a case study
- Author
-
Carlo Storelli, Tiziano Verri, Amilcare Barca, and Alessandro Romano
- Subjects
Genetics ,Physiology ,GenBank ,Peptide transporter 1 ,biology.protein ,Transporter ,Biology ,Membrane transport ,Adaptation ,biology.organism_classification ,Gene ,Zebrafish ,Solute carrier family - Abstract
Human genes for passive, ion-coupled transporters and exchangers are included in the so-called solute carrier (SLC) gene series, to date consisting of 52 families and 398 genes. Teleost fish genes for SLC proteins have also been described in the last two decades, and catalogued in preliminary SLC-like form in 50 families and at least 338 genes after systematic GenBank database mining (December 2010–March 2011). When the kinetic properties of the expressed proteins are studied in detail, teleost fish SLC transporters always reveal extraordinary ‘molecular diversity’ with respect to the mammalian counterparts, which reflects peculiar adaptation of the protein to the physiology of the species and/or to the environment where the species lives. In the case of the H+–oligopeptide transporter PEPT1(SLC15A1), comparative analysis of diverse teleost fish orthologs has shown that the protein may exhibit very eccentric properties in terms of pH dependence (e.g. the adaptation of zebrafish PEPT1 to alkaline pH), temperature dependence (e.g. the adaptation of icefish PEPT1 to sub-zero temperatures) and/or substrate specificity (e.g. the species-specificity of PEPT1 for the uptake of l-lysine-containing peptides). The revelation of such peculiarities is providing new contributions to the discussion on PEPT1 in both basic (e.g. molecular structure–function analyses) and applied research (e.g. optimizing diets to enhance growth of commercially valuable fish).
- Published
- 2013
9. Comparative Analysis and Functional Mapping of SACS Mutations Reveal Novel Insights into Sacsin Repeated Architecture
- Author
-
Maria Fulvia de Leva, Carlo Storelli, Amilcare Barca, Alessandro Romano, Filippo M. Santorelli, Tiziano Verri, Alessandra Tessa, Fabiana Fattori, Alessandra Terracciano, Romano, Alessandro, Alessandra, Tessa, Barca, Amilcare, Fabiana, Fattori, Maria Fulvia de, Leva, Alessandra, Terracciano, Storelli, Carlo, Filippo Maria, Santorelli, and Verri, Tiziano
- Subjects
Mutant ,Sequence alignment ,Biology ,medicine.disease_cause ,Polymorphism, Single Nucleotide ,Exon ,Phylogenetics ,Genetics ,medicine ,Humans ,Spinocerebellar Ataxias ,RNA, Messenger ,functional mapping of human mutations ,Heat-Shock Proteins ,Phylogeny ,Research Articles ,Genetics (clinical) ,Mutation ,Neurodegeneration ,neurodegeneration ,Chromosome Mapping ,Computational Biology ,functional mapping of human mutation ,Exons ,Sequence Analysis, DNA ,comparative protein analysis ,medicine.disease ,protein domain architecture ,Phenotype ,repeated domain ,autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) ,Muscle Spasticity ,repeated domains ,sacsin ,Spinocerebellar ataxia ,autosomal recessive spastic ataxia of Charlevoix–Saguenay (ARSACS) ,comparative protein analysi ,Sequence Alignment ,SACS - Abstract
Autosomal recessive spastic ataxia of Charlevoix–Saguenay (ARSACS) is a neurological disease with mutations in SACS, encoding sacsin, a multidomain protein of 4,579 amino acids. The large size of SACS and its translated protein has hindered biochemical analysis of ARSACS, and how mutant sacsins lead to disease remains largely unknown. Three repeated sequences, called sacsin repeating region (SRR) supradomains, have been recognized, which contribute to sacsin chaperone-like activity. We found that the three SRRs are much larger (≥1,100 residues) than previously described, and organized in discrete subrepeats. We named the large repeated regions Sacsin Internal RePeaTs (SIRPT1, SIRPT2, and SIRPT3) and the subrepeats sr1, sr2, sr3, and srX. Comparative analysis of vertebrate sacsins in combination with fine positional mapping of a set of human mutations revealed that sr1, sr2, sr3, and srX are functional. Notably, the position of the pathogenic mutations in sr1, sr2, sr3, and srX appeared to be related to the severity of the clinical phenotype, as assessed by defining a severity scoring system. Our results suggest that the relative position of mutations in subrepeats will variably influence sacsin dysfunction. The characterization of the specific role of each repeated region will help in developing a comprehensive and integrated pathophysiological model of function for sacsin.
- Published
- 2013
10. A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of D-glucose and choline transport systems
- Author
-
Oreste Acuto, Giorgio Semenza, Martin Müller, Carlo Storelli, Heini Murer, and Markus Kessler
- Subjects
Brush border ,Biophysics ,Biological Transport, Active ,In Vitro Techniques ,Cell Fractionation ,Biochemistry ,Choline ,Microvillus membrane ,Sucrase-isomaltase complex ,chemistry.chemical_compound ,Cricetinae ,Intestine, Small ,Electrochemistry ,Animals ,Chemical Precipitation ,HEPES ,Chromatography ,Microvilli ,Vesicle ,Cell Membrane ,Sodium ,Cell Biology ,Rats ,Kinetics ,Glucose ,Membrane ,chemistry ,Calcium ,Rabbits ,Choline transport ,Sucrase-isomaltase - Abstract
We have worked out a simplification of the procedure described by Schmitz et al. (Biochim. Biophys. Acta (1973) 323, 98--112) for the preparation of brush border membranes from small intestine. The procedure ultimately adopted is simple, rapid, does not necessarily require scraping and can be started from fresh or frozen material. It can be scaled up easily, allowing a quick production of large amounts of brush border membrane vesicles. These vesicles prove to be excellently suited for transport studies, as suggested by our measurements of D-glucose transport. Using these vesicles, the mode of choline transport across the brush border membrane was also investigated. Choline transport was found to occur by a saturable component with a Km of 83 +/- 4 micrometer (at 20 degrees C) and by a non-saturable component. It is independent of the presence of Na+ and appears to be non-electrogenic.
- Published
- 2016
11. Antitumor activity of the dietary diterpene carnosol against a panel of human cancer cell lines
- Author
-
Angelo Santino, Michele Maffia, Carlo Storelli, Silvana Leo, Simona Bettini, Daniele Vergara, Andrea Tinelli, Pasquale Simeone, Ludovico Valli, Vergara, Daniele, Simeone, P, Bettini, S, Tinelli, A, Valli, Ludovico, Storelli, C, Leo, S, Santino, A, Maffia, Michele, Daria, Vergara, Pasquale, Simeone, Bettini, Simona, Andrea, Tinelli, Carlo, Storelli, Silvana, Leo, and Angelo, Santino
- Subjects
ROSEMARY ,Cell Survival ,Phytochemicals ,INHIBITION ,Antineoplastic Agents ,Pharmacology ,Carnosol ,OVARIAN-CANCER ,chemistry.chemical_compound ,Cell Line, Tumor ,Cell Adhesion ,TUMORIGENESIS ,Humans ,diterpenecarnosol ,Viability assay ,Epithelial–mesenchymal transition ,TRANSCRIPTION ,Cell Proliferation ,biology ,Biological activity ,General Medicine ,EPITHELIAL-MESENCHYMAL TRANSITION ,Fibronectin ,MEDITERRANEAN DIET ,chemistry ,Cell culture ,Cancer cell ,Abietanes ,biology.protein ,Curcumin ,MCF-7 Cells ,Caco-2 Cells ,Antitumor activity ,OFFICINALIS ,Food Science - Abstract
Dietary phytochemicals found in vegetables and fruits consist of a wide variety of biologically active compounds with anti-carcinogenic activity. The aim of this study was to evaluate the antigrowth activity of carnosol, a dietary diterpene, as a single agent or in combination with other dietary phytochemicals or chemotherapeutic drugs against a panel of tumor cell lines. Carnosol decreased cell viability in human breast, ovarian, and intestinal tumor cell lines, and inhibited cancer cell adhesion on fibronectin and growth of cancer cells in suspension. Carnosol also inhibited EGF-induced epithelial mesenchymal transition in ovarian cancer cells. The combination treatment with other dietary phytochemicals increased the anti-proliferative activity of carnosol. The combination with curcumin resulted in a synergistic reduction of vitality in SKOV-3 and MDA-231 cells and potently inhibited viability of primary cancer cells isolated from the pleural fluid or ascites of patients with metastatic cancers. These results provide additional evidence about the anticancer role of carnosol and its potential in blocking the growth of tumor cells. © 2014 the Partner Organisations.
- Published
- 2014
12. Multiple anti-inflammatory and anti-atherosclerotic properties of red wine polyphenolic extracts
- Author
-
Nadia Calabriso · Egeria Scoditti · Marika Massaro · Mariangela Pellegrino · Carlo Storelli · Ilaria Ingrosso · Giovanna Giovinazzo · Maria Annunziata Carluccio
- Subjects
Macrophage colony-stimulsting factor ,food and beverages ,intercellular adhesion molecule-I ,Monocyte chemoactrant protein-I ,Vascular cell adhesion molecule-I ,E-Selectin - Abstract
Purpose: The aim of the study was to evaluate the vascular anti-inflammatory effects of polyphenolic extracts from two typical South Italy red wines, the specific contribution of individual polyphenols and the underlying mechanisms of action. Methods: Human endothelial cells were incubated with increasing concentrations (1-50 ?g/mL) of Primitivo and Negroamaro polyphenolic extracts (PWPE and NWPE, respectively) or pure polyphenols (1-25 ?mol/L), including hydroxycinnamic acids (p-coumaric, caffeic and caftaric acids), flavonols (kaempferol, quercetin, myricetin) or stilbenes (trans-resveratrol, trans-piceid) before stimulation with lipopolysaccharide. Through multiple assays, we analyzed the endothelial-monocyte adhesion, the endothelial expression of adhesion molecules (ICAM-1, VCAM-1 and E-Selectin), monocyte chemoattractant protein-1 (MCP-1) and macrophage colony-stimulating factor (M-CSF), as well as ROS intracellular levels and the activation of NF-?B and AP-1. Results: Both PWPE and NWPE, already at 1 ?g/mL, inhibited monocyte adhesion to stimulated endothelial cells, a key event in triggering vascular inflammation. They down-regulated the expression of adhesion molecules, ICAM-1, VCAM-1, E-Selectin, as well as MCP-1 and M-CSF, at mRNA and protein levels. All polyphenols reduced intracellular ROS, and everything, except caftaric acid, inhibited the endothelial expression of adhesion molecules and MCP-1, although with different potency. Flavonols and resveratrol significantly reduced also the endothelial expression and release of M-CSF. The decrease in endothelial inflammatory gene expression was related to the inhibition of NF-?B and AP-1 activation but not to intracellular oxidative stress. Conclusions: This study showed multiple anti-inflammatory and anti-atherosclerotic properties of red wine polyphenolic extracts and indentified specific bioactive polyphenols which could counteract inflammatory diseases including atherosclerosis.
- Published
- 2016
- Full Text
- View/download PDF
13. Therapeutic potential of the dual peroxisome proliferator activated receptor (PPAR)α/γ agonist aleglitazar in attenuating TNF-α-mediated inflammation and insulin resistance in human adipocytes
- Author
-
Marika Massaro, Egeria Scoditti, Raffaele De Caterina, Nadia Calabriso, Mariangela Pellegrino, Maria Annunziata Carluccio, Matthew Blake Wright, Martin Wabitsch, and Carlo Storelli
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Cell Survival ,Adipose tissue ,Peroxisome proliferator-activated receptor ,Thiophenes ,p38 Mitogen-Activated Protein Kinases ,PPAR agonist ,Cell Line ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Movement ,Internal medicine ,medicine ,Adipocytes ,Humans ,Hypoglycemic Agents ,PPAR alpha ,Oxazoles ,Chemokine CCL2 ,Pharmacology ,chemistry.chemical_classification ,Inflammation ,Aleglitazar ,biology ,Adiponectin ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Lipid Metabolism ,IRS1 ,Chemokine CXCL10 ,PPAR gamma ,Insulin receptor ,030104 developmental biology ,Endocrinology ,Glucose ,chemistry ,Gene Expression Regulation ,biology.protein ,Insulin Resistance ,Rosiglitazone ,medicine.drug - Abstract
Adipose tissue inflammation is a mechanistic link between obesity and its related sequelae, including insulin resistance and type 2 diabetes. Dual ligands of peroxisome proliferator activated receptor (PPAR)α and γ, combining in a single molecule the metabolic and inflammatory-regulatory properties of α and γ agonists, have been proposed as a promising therapeutic strategy to antagonize adipose tissue inflammation. Here we investigated the effects of the dual PPARα/γ agonist aleglitazar on human adipocytes challenged with inflammatory stimuli. Human Simpson-Golabi-Behmel syndrome (SGBS) adipocytes were treated with aleglitazar or - for comparison - the selective agonists for PPARα or γ fenofibrate or rosiglitazone, respectively, for 24h before stimulation with TNF-α. Aleglitazar, at concentrations as low as 10nmol/L, providing the half-maximal transcriptional activation of both PPARα and PPARγ, reduced the stimulated expression of several pro-inflammatory mediators including interleukin (IL)-6, the chemokine CXC-L10, and monocyte chemoattractant protein (MCP)-1. Correspondingly, media from adipocytes treated with aleglitazar reduced monocyte migration, consistent with suppression of MCP-1 secretion. Under the same conditions, aleglitazar also reversed the TNF-α-mediated suppression of insulin-stimulated ser473 Akt phosphorylation and decreased the TNF-α-induced ser312 IRS1 phosphorylation, two major switches in insulin-mediated metabolic activities, restoring glucose uptake in insulin-resistant adipocytes. Such effects were similar to those obtainable with a combination of single PPARα and γ agonists. In conclusion, aleglitazar reduces inflammatory activation and dysfunction in insulin signaling in activated adipocytes, properties that may benefit diabetic and obese patients. The effect of aleglitazar was consistent with dual PPARα and γ agonism, but with no evidence of synergism.
- Published
- 2016
14. Changes in hormonal profile, gonads and sperm quality of Argyrosomus regius (Pisces, Scianidae) during the first sexual differentiation and maturation
- Author
-
Loredana Zilli, Roberta Schiavone, Sebastiano Vilella, Carlo Storelli, Schiavone, Roberta, Zilli, Loredana, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Male ,medicine.medical_specialty ,Sex Differentiation ,Argyrosomus regius ,Andrology ,Food Animals ,Internal medicine ,Reproductive biology ,medicine ,Animals ,Sexual maturity ,Testosterone ,Sexual Maturation ,Gonads ,Small Animals ,Sperm motility ,Sexual differentiation ,Estradiol ,biology ,Equine ,biology.organism_classification ,Spermatozoa ,Sperm ,Hormones ,Perciformes ,Gonadosomatic Index ,Endocrinology ,Meagre, Sex steroids, Gonadal histology, CASA ,Sperm Motility ,Female ,Animal Science and Zoology - Abstract
In the present study, sexual gonadal differentiation and first sexual maturation of Meagre ( Argyrosomus regius ) was studied, based upon the annual changes in gonadosomatic index (GSI), gonadal histology, and the plasma steroid hormones, testosterone (T), 11-ketotestosterone (11-KT), and estradiol (E2). In addition, spermatozoa characteristics were evaluated by measuring sperm motility and morphology. Results demonstrated that Meagre completes sex differentiation at 10 to 12 mo of age, and are group-synchronous spawners, which reach puberty at 2 (mean length 26.8 ± 0.7 cm, mean weight 920 ± 75 g; N = 10) and 3 (mean length 35.8 ± 0.8 cm, mean weight 1610 ± 89 g; N = 10) years of age for males and females, respectively. In males, during the sex differentiation period, T levels were significantly higher with respect to those of 11-KT; this suggests that T has a key role in the early phases of the sex differentiation. During the spawning season an increase in plasma concentrations of all hormones was observed with 11-KT levels being significantly higher that those of T. In females, during the sex differentiation period, there was an increase in E2 plasma levels, while during the first spawning season, a significant increase of T and E2 levels were measured. Regarding sperm characteristics, the measured curvilinear velocity (VCL) and straight-linear velocity (VSL), resulted in the same order of magnitude with respect to those measured in other marine fish, while the average path velocity (VAP) was similar to that measured in the European Eel. The head of Meagre spermatozoa presents as oval shaped with a surface area of approximately 3.66 μm 2 and a perimeter of approximately 6.65 μm. All these findings represent an important basis for further investigation on the reproductive biology of this specie and may assist the farmers to improve seed production in aquaculture.
- Published
- 2012
15. Functional expression of SLC15 peptide transporters in rat thyroid follicular cells
- Author
-
Hannelore Daniel, Gabor Kottra, Alessandro Romano, Carlo Storelli, Amilcare Barca, Tiziano Verri, Romano, A., Barca, A., Kottra, G., Daniel, H., Storelli, Carlo, and Verri, Tiziano
- Subjects
β-Ala-Lys-N-7-amino-4-methyl-coumarin-3-acetic acid ,endocrine system diseases ,medicine.medical_treatment ,SoLute Carrier 15 family ,Thyroid Gland ,Thyrotropin ,thyroglobulin ,Biochemistry ,Rats, Sprague-Dawley ,chemistry.chemical_compound ,0302 clinical medicine ,Endocrinology ,beta-Ala-Lys-Nepsilon-7-amino-4-methyl-coumarin-3-acetic acid ,Ala-Lys-AMCA ,Protein Isoforms ,Tg ,thyroid follicular cell (thyrocyte) ,0303 health sciences ,Symporters ,medicine.diagnostic_test ,Chemistry ,Dipeptides ,Intracellular ,endocrine system ,Proteolysis ,Nerve Tissue Proteins ,Peptide Transporter 1 ,Cell Line ,03 medical and health sciences ,medicine ,Animals ,RNA, Messenger ,Molecular Biology ,030304 developmental biology ,PC Cl3 cell ,Dipeptide ,Thyroid hormone receptor ,SLC15 ,Membrane Transport Proteins ,Rats ,Solute carrier family ,carnosine ,Peptide transport ,Cell culture ,PC Cl3 cells ,Thyroglobulin ,030217 neurology & neurosurgery ,Solute Carrier 15 family - Abstract
International audience; Peptide transport and expression of SoLute Carrier 15 (SLC15) peptide transporters was assessed in rat thyroid tissue and a rat thyroid cell line (PC Cl3 cells). Peptide transport was studied by monitoring the uptake of the fluorophore-conjugated dipeptide β-Ala-Lys-N-7-amino-4-methyl-coumarin-3-acetic acid (Ala-Lys-AMCA). Expression of SLC15-specific mRNA transcripts was analyzed by RT-PCR. Of the two SLC15 transporters expressed in thyroid follicular cells, namely PEPT2 (SLC15A2) and PHT1 (SLC15A4), only PEPT2 was involved in peptide transport at the plasma membrane, with PHT1 most likely being intracellular. Interestingly, at the mRNA level PEPT2 was up-regulated under TSH stimulation. These findings represent the first evidence that peptide transport occurs in thyroid follicular cells. SLC15 transporters could participate to recycling of peptides derived from extracellular and lysosomal thyroglobulin proteolysis, both essential steps for thyroid hormone synthesis.
- Published
- 2010
16. Comparison among Different Gilthead Sea Bream (Sparus aurata) Farming Systems: Activity of Intestinal and Hepatic Enzymes and 13C-NMR Analysis of Lipids
- Author
-
Paride Papadia, Sandra Angelica De Pascali, Francesco Paolo Fanizzi, Laura Del Coco, Carlo Storelli, V. Zonno, Giorgia Bressani, Del Coco, L., Papadia, Paride, DE PASCALI, SANDRA ANGELICA, Bressani, Giorgia, Storelli, Carlo, Zonno, Vincenzo, and Fanizzi, Francesco Paolo
- Subjects
Magnetic Resonance Spectroscopy ,Muscle Proteins ,lcsh:TX341-641 ,Aquaculture ,Aminopeptidase ,Article ,food authenticity ,13C NMR profiling ,Animals ,Food science ,nutritional physiological state ,Muscle, Skeletal ,Leucyl aminopeptidase ,chemistry.chemical_classification ,Carbon Isotopes ,Nutrition and Dietetics ,biology ,gilthead sea bream (Sparus aurata) ,Lipids ,Sea Bream ,Intestines ,Cholesterol ,Liver ,Biochemistry ,chemistry ,Alpha-glucosidase ,Saturated fatty acid ,biology.protein ,Alkaline phosphatase ,rearing systems ,Leucine ,Maltase ,lcsh:Nutrition. Foods and food supply ,PUFA ,Food Science ,Polyunsaturated fatty acid - Abstract
In order to evaluate differences in general health and nutritional values of gilthead sea bream (Sparus aurata), the effects of semi-intensive, land-based tanks and sea-cages intensive rearing systems were investigated, and results compared with captured wild fish. The physiological state was determined by measuring the activity of three different intestinal digestive enzymes: alkaline phosphatase (ALP), leucine aminopeptidase (LAP) and maltase; and the activity of the hepatic ALP. Also, the hepatic content in protein, cholesterol, and lipid were assessed. 13C-NMR analysis for qualitative and quantitative characterization of the lipid fraction extracted from fish muscles for semi-intensive and land based tanks intensive systems was performed. The lipid fraction composition showed small but significant differences in the monounsaturated/saturated fatty acid ratio, with the semi-intensive characterized by higher monounsaturated and lower saturated fatty acid content with respect to land based tanks intensive rearing system.
- Published
- 2009
17. Evidence for the Involvement of Aquaporins in Sperm Motility Activation of the Teleost Gilthead Sea Bream (Sparus aurata)1
- Author
-
François Chauvigné, Carlo Storelli, Loredana Zilli, Roberta Schiavone, Joan Cerdà, and Sebastiano Vilella
- Subjects
Osmotic concentration ,medicine.diagnostic_test ,urogenital system ,Xenopus ,Motility ,Aquaporin ,Cell Biology ,General Medicine ,Anatomy ,Biology ,biology.organism_classification ,Sperm ,Cell biology ,Reproductive Medicine ,Western blot ,medicine ,Heterologous expression ,Sperm motility - Abstract
The expression of aquaporins in the spermatozoa of the marine teleost gilthead sea bream (Sparus aurata) and their involvement in the motility activation process were investigated. Sperm motility was activated by a hyperosmotic shock, but it was completely inhibited by 10 μM HgCl2, such inhibition being partially recovered by beta-mercaptoethanol (ME). Conventional RT-PCR using primers specific for S. aurata aquaglyceroporin (glp) and aquaporin 1a (aqp1a) demonstrated the presence of both mRNAs in spermatozoa. Heterologous expression in Xenopus laevis oocytes showed that 10 and 100 μM HgCl2 equally inhibited water and solute transport through S. aurata aquaporin 1a and S. aurata aquaglyceroporin, but treatment with ME only recovered aquaporin 1a-mediated water permeability. Western blot analysis using isoform-specific antisera on protein extracts from spermatozoa revealed bands that corresponded to the predicted molecular mass of S. aurata aquaglyceroporin (31 kDa) and S. aurata aquaporin 1a (28 kDa). The...
- Published
- 2009
18. Anti-apoptotic effects of protein kinase C-δ and c-fos in cisplatin-treated thyroid cells
- Author
-
Loredana Urso, Santo Marsigliante, Nadia Calabriso, Antonella Muscella, Alessio Rochira, Carla Vetrugno, and Carlo Storelli
- Subjects
Pharmacology ,Cisplatin ,MAPK/ERK pathway ,Small interfering RNA ,Kinase ,Biology ,Molecular biology ,Mitogen-activated protein kinase ,medicine ,Cancer research ,biology.protein ,Viability assay ,Protein kinase A ,Protein kinase C ,medicine.drug - Abstract
Background and purpose: We showed previously that cisplatin inititates a signalling pathway mediated by PKC-δ/extracellular signal-regulated kinase (ERK), important for maintaining viability in PC Cl3 thyroid cells. The studies described herein examined whether c-fos was associated with cisplatin resistance and the signalling link between c-fos and PKC-δ/ERK. Experimental approach: Cells were treated with various pharmacological inhibitors of PKCs and ERK, or were depleted of c-fos, PKC-δ, PKC-e and caspase-3 by small interfering RNA (siRNA), then incubated with cisplatin and cytotoxicity assessed. Key results: Cisplatin provokes the induction of c-fos and the activation of conventional PKC-β, and novel PKC-δ and -e. The cisplatin-provoked c-fos induction was decreased by Go6976, a PKC-β inhibitor; by siRNA for PKC-δ- but not that for PKC-e or by PD98059, a mitogen-activated protein kinase/ERK kinase inhibitor. Expression of c-fos was abolished by GF109203X, an inhibitor of all PKC isoforms, or by PD98059 plus Go6976 or by PKC-δ-siRNA plus Go6976. When c-fos expression was blocked by siRNA, cisplatin cytotoxicity was strongly enhanced with increased caspase-3 activation. In PKC-δ-depleted cells treated with cisplatin, caspase-3 activation was increased and cell viability decreased. In these PKC-δ-depleted cells, PD98059 did not affect caspase-3 activation. Conclusions and implications: In PC Cl3 cells, in the cell signalling pathways that lead to cisplatin resistance, PKC-δ controls ERK activity and, together with PKC-β, also the induction of c-fos. Hence, the protective role of c-fos in thyroid cells has the potential to provide new opportunities for therapeutic intervention.
- Published
- 2009
19. A rapid and inexpensive method to assay transport of short chain peptides across intestinal brush-border membrane vesicles from the European eel (Anguilla anguilla)
- Author
-
Carlo Storelli, Ivar Rønnestad, Amilcare Barca, Snorre Bakke, Antonio Danieli, Michele Maffia, Alessandro Romano, Tiziano Verri, Verri, Tiziano, Danieli, A., Bakke, S., Romano, A., Barca, A., Ronnestad, I., Maffia, Michele, and Storelli, Carlo
- Subjects
Membrane potential ,chemistry.chemical_classification ,Brush border ,Chemistry ,Vesicle ,Depolarization ,Peptide ,PepT1 ,Aquatic Science ,Biochemistry ,In vivo ,Peptide transport ,DiS-C2(5) ,di/tripeptide transport ,Slc15A1 ,membrane potential ,Cotransporter ,Cyanine dye - Abstract
Membrane potential depolarization due to electrogenic peptide transport activity was examined in eel (Anguilla anguilla) intestinal brush-border membrane vesicles (BBMV) by monitoring the fluorescence quenching of the voltage-sensitive dye 3,3'-diethylthiadicarbocyanine iodide. Our experimental approach consisted of generating an internal negative membrane potential mimicking in vivo conditions and measuring membrane potential depolarization due to different extravesicular dipeptides. Peptide-dependent membrane potential depolarization was observed in both the presence and absence of extravesicular Na+ and was inhibited by diethylpyrocarbonate, which is consistent with the involvement of electrogenic, Na+-independent, H+-dependent peptide transport activity. Kinetic analysis indicated that peptide-dependent membrane potential depolarization is a saturable process (K-m,K-app similar to 1.5 mmol L-1) and that within the 0.1-10 mmol L-1 peptide range a single carrier system is involved in the transport process. Our results suggest that a peptide transport activity, kinetically resembling the PepT1(Slc15A1)-type-mediated H+/peptide cotransport action, can be monitored in eel intestinal BBMV using an easy and inexpensive fluorescence assay.
- Published
- 2008
20. Molecular Mechanisms Determining Sperm Motility Initiation in Two Sparids (Sparus aurata and Lithognathus mormyrus)
- Author
-
Loredana Zilli, Sebastiano Vilella, Carlo Storelli, Roberta Schiavone, Zilli, L, Schiavone, R, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Male ,Proteome ,Motility ,Biology ,sperm ,Dephosphorylation ,Lithognathus mormyrus ,Cyclic AMP ,Animals ,Protein phosphorylation ,sperm motility and transport ,sparid ,Sperm motility ,Molecular mass ,phosphorylation ,Osmolar Concentration ,motility initiation ,Cell Biology ,General Medicine ,Anatomy ,Cyclic AMP-Dependent Protein Kinases ,Spermatozoa ,Sperm ,Sea Bream ,Cell biology ,Reproductive Medicine ,Potassium ,Sperm Motility ,Phosphorylation ,Calcium ,signal transduction - Abstract
Molecular mechanisms involved in sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus) have been studied. Our comparative study demonstrates that osmolality is the key signal in sperm motility activation in both species, whereas K(+) and Ca(2+) do not have any role. The straight-line velocity that resulted, however, was significantly different when measured in sperm activated with non-ionic and/or calcium-free solutions with respect to that measured in seawater-activated sperm. In both species, motility initiation depends on cAMP-dependent protein phosphorylation. The phosphorylation/dephosphorylation patterns that resulted in gilthead and striped sea bream were quite different. In gilthead sea bream, the phosphorylated proteins have molecular weights of 174, 147, 138, 70, and 9-15 kDa, whereas the dephosphorylated proteins have molecular weights of 76, 57, and 33 kDa. In striped sea bream, phosphorylation after sperm motility activation occurred on proteins of 174, 147, 103, 96, 61, 57, and 28 kDa, whereas only one protein of 70 kDa resulted from dephosphorylation. Matrix-assisted laser desorption ionization-time of flight analyses allowed identification of the following proteins: In gilthead sea bream, the 9-15 kDa proteins that were phosphorylated after motility activation include an A-kinase anchor protein (AKAP), an acetyl-coenzyme A synthetase, and a protein phosphatase inhibitor, and in striped sea bream, 103- and 61-kDa proteins that were phosphorylated after motility activation were identified as a phosphatase (myotubularin-related protein 1) and a kinase (DYRK3), respectively.
- Published
- 2008
21. Identification by proteome analysis of muscle proteins in sea bream (Sparus aurata)
- Author
-
Roberta Schiavone, Sebastiano Vilella, Loredana Zilli, Carlo Storelli, Schiavone, Roberta, Zilli, Loredana, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
MALDI-TOF ,Gel electrophoresis ,Two-dimensional gel electrophoresis ,Myosin light-chain kinase ,Chromatography ,2D-electrophoresi ,General Chemistry ,fish muscle ,Biology ,Mass spectrometry ,Biochemistry ,Tropomyosin ,Industrial and Manufacturing Engineering ,Electrophoresis ,Proteome ,biology.protein ,protein ,sea bream ,Parvalbumin ,Food Science ,Biotechnology - Abstract
The present paper reported postmortem changes in muscle proteins on sea bream (Sparus aurata), during ice storage. The postmortem evolution of protein patterns in farmed sea bream was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimension electrophoresis (2DE). Matrix associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was used for identification of proteins. Results of the bio-informatics analysis demonstrate that: (1) the normalized volume of skeletal alpha-actin and tropomyosin is not affected by the time of storage; (2) the normalized volumes of myosin light chain 3 and of major histocompatibility complex (MHC) class II beta1 increase; (3) the normalized volumes of Sec 13-like and parvalbumin significantly decrease.
- Published
- 2008
22. THE PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR(PPAR)GAMMA AGONIST ROSIGLITAZONE (RSG) INHIBITS CYCLOOXYGENASE(COX)-2 EXPRESSION BY SUPPRESSING PKCAPLHA AND CREB ACTIVATION
- Author
-
Egeria Scoditti, Carlo Storelli, R. De Caterina, Alessandro Distante, MA Carluccio, and Marika Massaro
- Subjects
chemistry.chemical_classification ,biology ,chemistry ,Cancer research ,biology.protein ,medicine ,Peroxisome proliferator-activated receptor ,Hematology ,Cyclooxygenase ,CREB ,Rosiglitazone ,PPAR agonist ,medicine.drug - Published
- 2007
23. HYDROXYTYROSOL, THE MAJOR OLIVE OIL PHENOLIC ANTIOXIDANT, REGULATES EXPRESSION AND ACTIVITY OF MATRIX METALLOPROTEINASE(MMP)-9 IN HUMAN MONOCYTOID CELLS - POSSIBLE CONTRIBUTION TO PLAQUE STABILITY
- Author
-
Marika Massaro, R. De Caterina, MA Carluccio, A. Nestola, Carlo Storelli, Alessandro Distante, and Egeria Scoditti
- Subjects
Phenolic antioxidant ,chemistry.chemical_compound ,Biochemistry ,Chemistry ,Hydroxytyrosol ,Hematology ,Monocytoid Cells ,Matrix metalloproteinase ,Olive oil - Published
- 2007
24. THE OMEGA-3 FATTY ACID DOCOSAHEXAENOATE (DHA) REDUCES CYCLOOXYGENASE(COX)-2 INDUCTION BY INHIBITING NADP(H) OXIDASE AND PKC[epsiv]: POSSIBLE INVOLVEMENT OF 15-LIPOXIGENASE(LO)-1
- Author
-
MA Carluccio, Egeria Scoditti, Carlo Storelli, Alessandro Distante, Marika Massaro, and R. De Caterina
- Subjects
Oxidase test ,Biochemistry ,biology ,Chemistry ,biology.protein ,Hematology ,Cyclooxygenase ,Omega 3 fatty acid ,Protein kinase C - Published
- 2007
25. Extra virgin olive oil rich in polyphenols modulates VEGF-induced angiogenic responses by preventing NADPH oxidase activity and expression
- Author
-
Simona D'Amore, Nadia Calabriso, Maria Annunziata Carluccio, Marika Massaro, Raffaele De Caterina, Giuseppe Palasciano, Egeria Scoditti, Mariangela Pellegrino, Antonio Gnoni, and Carlo Storelli
- Subjects
0301 basic medicine ,Vascular Endothelial Growth Factor A ,Antioxidant ,Angiogenesis ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Clinical Biochemistry ,Matrix metalloproteinase ,medicine.disease_cause ,Biochemistry ,03 medical and health sciences ,medicine ,Human Umbilical Vein Endothelial Cells ,Humans ,Molecular Biology ,Olive Oil ,Nutrition and Dietetics ,NADPH oxidase ,biology ,Neovascularization, Pathologic ,food and beverages ,NOX4 ,NADPH Oxidases ,Polyphenols ,Vascular endothelial growth factor A ,030104 developmental biology ,Polyphenol ,biology.protein ,Endothelium, Vascular ,Oxidative stress - Abstract
Previous studies have shown the antiinflammatory, antioxidant and antiangiogenic properties by pure olive oil polyphenols; however, the effects of olive oil phenolic fraction on the inflammatory angiogenesis are unknown. In this study, we investigated the effects of the phenolic fraction (olive oil polyphenolic extract, OOPE) from extra virgin olive oil and related circulating metabolites on the VEGF-induced angiogenic responses and NADPH oxidase activity and expression in human cultured endothelial cells. We found that OOPE (1-10 μg/ml), at concentrations achievable nutritionally, significantly reduced, in a concentration-dependent manner, the VEGF-induced cell migration, invasiveness and tube-like structure formation through the inhibition of MMP-2 and MMP-9. OOPE significantly (P
- Published
- 2015
26. Transcriptome-based identification of new anti-anti-inflammatory and vasodilating properties of the n-3 fatty acid docosahexaenoic acid in vascular endothelial cell under proinflammatory conditions
- Author
-
Rosanna Martinelli, Marika Massaro, Carlo Storelli, Liborio Stuppia, Raffaele De Caterina, Valentina Gatta, Nadia Calabriso, Tonia Buonomo, Maria Annunziata Carluccio, Egeria Scoditti, and Mariangela Pellegrino
- Subjects
Genetics and Molecular Biology (all) ,Endothelium ,Endothelial cells ,lcsh:Medicine ,Microarray ,Pharmacology ,Biology ,Biochemistry ,Proinflammatory cytokine ,Transcriptome ,Agricultural and Biological Sciences (all) ,Medicine (all) ,DNA-binding proteins ,Gene expression ,medicine ,Gene regulation ,Genetic interference ,Small interfering RNAs ,lcsh:Science ,Regulation of gene expression ,chemistry.chemical_classification ,Multidisciplinary ,lcsh:R ,Fatty acid ,3. Good health ,Gene expression profiling ,medicine.anatomical_structure ,chemistry ,Docosahexaenoic acid ,Immunology ,lcsh:Q ,Research Article - Abstract
Scope High intakes of n-3 fatty acids exert anti-inflammatory effects and cardiovascular protection, but the underlying molecular basis is incompletely defined. By genome-wide analysis we searched for novel effects of docosahexaenoic acid (DHA) on gene expression and pathways in human vascular endothelium under pro-inflammatory conditions. Methods and Results Human umbilical vein endothelial cells were treated with DHA and then stimulated with interleukin(IL)-1β. Total RNA was extracted, and gene expression examined by DNA microarray. DHA alone altered the expression of 188 genes, decreasing 92 and increasing 96. IL-1β changed the expression of 2031 genes, decreasing 997 and increasing 1034. Treatment with DHA before stimulation significantly affected the expression of 116 IL-1β-deregulated genes, counter-regulating the expression of 55 genes among those decreased and of 61 among those increased. Functional and network analyses identified immunological, inflammatory and metabolic pathways as the most affected. Newly identified DHA-regulated genes are involved in stemness, cellular growth, cardiovascular system function and cancer, and included cytochrome p450 4F2(CYP4F2), transforming growth factor(TGF)-β2, Cluster of Differentiation (CD)47, caspase recruitment domain(CARD)11 and phosphodiesterase(PDE)5α. Conclusions Endothelial exposure to DHA regulates novel genes and related pathways. Such unbiased identification should increase our understanding of mechanisms by which n-3 fatty acids affect human diseases.
- Published
- 2015
27. miR-15b and miR-21 as Circulating Biomarkers for Diagnosis of Glioma
- Author
-
Oscar Fernando D'Urso, Valeria Mezzolla, Pietro I. D’Urso, Cosimo Damiano Gianfreda, Santo Marsigliante, Carlo Storelli, Ivo D’Urso, Pietro, Fernando, Oscar D’Urso, Damiano Gianfreda, Cosimo, Mezzolla, Valeria, Storelli, Carlo, and Marsigliante, Santo
- Subjects
Oncology ,medicine.medical_specialty ,Pathology ,Microarray ,Microarrays ,Biomarkers, Blood, Diagnosis, Glioma, Microarrays, miRNAs ,Disease ,medicine.disease_cause ,Article ,law.invention ,law ,Internal medicine ,Glioma ,Diagnosis ,microRNA ,Genetics ,medicine ,Genetics (clinical) ,Polymerase chain reaction ,business.industry ,medicine.disease ,Blood ,miRNAs ,Biomarker (medicine) ,DNA microarray ,Carcinogenesis ,business ,Biomarkers - Abstract
Malignant gliomas are lethal primary intracranial tumors. To date, little information on the role of deregulated genes in gliomas have been identified. As the involvement of miRNAs in the carcinogenesis is well known, we carried out a pilot study to identify, as potential biomarkers, differentially expressed microRNAs in blood samples of patients affected by glioma. We studied the miRNAs' expression, by means of microarray and Real-Time PCR, in 30 blood samples from glioma patients and in 82 blood samples of patients suffering from: (a) various neurological disorders (n=30), (b) primary B-lymphoma of the Central Nervous System (PCNSL, n=36) and (c) secondary brain metastases (n=16). By quantitative real time reverse-transcriptase polymerase chain reaction (qRT-PCR), we identified significantly increased levels of two candidate biomarkers, miR-15b and miR-21, in blood of patients affected by gliomas. ROC analysis of miR-15b biomarker levels allowed to differentiate patients with tumour from patients without glioma. Furthermore, combined expression analyses of miR15b and miR-21 distinguished between patients with and without glioma (90% sensitivity and 100% specificity). In addition, a decrement in the expression levels of miR-16 characterized glioblastomas compared to low grade and anaplastic gliomas. In conclusion, this pilot study suggest that it's possible to identify the disease state by meaning miR-15b and miR-21 markers in blood, while miR-16 can be used to distinguish glioblastoma from other grade gliomas. They can potentially be used as biomarkers for non-invasive diagnosis of gliomas; further studies are mandatory to confirm our preliminary findings.
- Published
- 2015
28. Angiotensin II does not stimulate proliferation of rat thyroid PC Cl3 cell line
- Author
-
Simona Romano, Antonella Muscella, Santo Marsigliante, Carlo Storelli, S., Romano, Muscella, Antonella, Storelli, Carlo, and Marsigliante, Santo
- Subjects
MAPK/ERK pathway ,medicine.medical_specialty ,Indoles ,Morpholines ,Endocrinology, Diabetes and Metabolism ,Thyroid Gland ,Cell Line ,Maleimides ,Phosphatidylinositol 3-Kinases ,Endocrinology ,Downregulation and upregulation ,Internal medicine ,medicine ,Animals ,Insulin ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Protein kinase A ,Protein kinase B ,Protein Kinase C ,Cell Proliferation ,Phosphoinositide-3 Kinase Inhibitors ,Flavonoids ,PC Cl3 cell ,PKB/Akt ,biology ,Kinase ,Cell growth ,Angiotensin II ,Cyclin-dependent kinase 2 ,Rats ,Enzyme Activation ,Ang II ,ERK ,cell proliferation ,Chromones ,biology.protein ,RNA Interference ,Proto-Oncogene Proteins c-akt ,Proto-Oncogene Proteins c-fos ,Cyclin-Dependent Kinase Inhibitor p27 ,Signal Transduction - Abstract
In PC Cl3 cells, a rat thyroid cell line, angiotensin (Ang II) activates the atypical protein kinase C-ζ (PKC-ζ) and the extracellular signal-regulated kinase (ERK) pathways. We here studied the Ang II effects on PC Cl3 cell proliferation. It was found that Ang II: (1) induced the phosphorylation of protein kinase B (PKB), (2) induced the growth-related early gene c-fos expression, (3) enhanced the cyclin E and p27kip expression, (4) had no effects on Cdk2, and (5) did not affect the transition from G0/G1 to S phase. Inhibition of phosphoinositide-3kinase by LY294002 further increased the effect of Ang II on p27kip induction, whilst PKCs inhibition by GF109203X decreased such effect. The role of PKC-ζ was recognized by the use of a synthetic myristoylated peptide with sequences based on the endogenous PKC-ζ pseudosubstrate and by PKC-ζ downregulation using the small interfering RNA (siRNA). Insulin had a replicating effect on PC Cl3 cells, induced the phosphorylation of PKB, decreased p27kip expression and had no effect on the PKC-ζ cytosol-to-membrane translocation. PC Cl3 cell proliferation was induced more potently by simultaneous stimulation with insulin and Ang II than by stimulation with insulin alone, and the effect on p27kip expression was similar to that obtained with insulin only. These observations demonstrate that in PC Cl3 cells Ang II causes a block in G1 phase, although both ERK and PKB pathways are activated, and this effect may be due to the upregulation of p27kip and PKC-ζ operativity.
- Published
- 2006
29. The sarcoplasmic–endoplasmic reticulum Ca2+ ATPase 2b regulates the Ca2+ transients elicited by P2Y2 activation in PC Cl3 thyroid cells
- Author
-
Carlo Storelli, Bruno Di Jeso, M. G. Elia, Santo Marsigliante, Antonella Muscella, Antonella Sonia Treglia, Luca Ulianich, L., Ulianich, Elia, Mg, Treglia, A, Muscella, Antonella, DI JESO, Bruno, Storelli, Carlo, and Marsigliante, Santo
- Subjects
medicine.medical_specialty ,Indoles ,Endocrinology, Diabetes and Metabolism ,ATPase ,Sarcoplasm ,Carbazoles ,Thyroid Gland ,Biological Transport, Active ,Uridine Triphosphate ,Cell Line ,Sarcoplasmic Reticulum Calcium-Transporting ATPases ,Maleimides ,Receptors, Purinergic P2Y2 ,Basal (phylogenetics) ,Adenosine Triphosphate ,Endocrinology ,P2Y2 ,SERCA ,Internal medicine ,medicine ,Animals ,Protein Kinase C ,Protein kinase C ,PC Cl3 cell ,Manganese ,biology ,Receptors, Purinergic P2 ,Endoplasmic reticulum ,Rats ,Enzyme Activation ,Sarcoplasmic Reticulum ,Membrane ,Microscopy, Fluorescence ,Barium ,Cell culture ,Permeability (electromagnetism) ,biology.protein ,Tetradecanoylphorbol Acetate ,Thapsigargin ,Calcium - Abstract
In PC Cl3 cells, a continuous, fully differentiated rat thyroid cell line, P2Y2 purinoceptor activation provoked a transient increase of [Ca2+]i, followed by a decreasing sustained phase. The α and β1 protein kinase C (PKC) inhibitor Gö6976 decreased the rate of decrement to the basal [Ca2+]i level and increased the peak of Ca2+ entry of the P2Y2-provoked Ca2+transients. These effects of Gö 6976 were not caused by an increased permeability of the plasma membrane, since the Mn2+ and Ba2+ uptake were not changed by Gö 6976. Similarly, the Na+/Ca2+ exchanger was not implicated, since the rate of decrement to the basal [Ca2+]i level was equally decreased in physiological and Na+-free buffers, in the presence of Gö 6976. On the contrary, the activity of the sarcoplasmic–endoplasmic reticulum Ca2+ATPase (SERCA) 2b was profoundly affected by Gö 6976 since the drug was able to completely inhibit the stimulation of the SERCA 2b activity elicited by P2-purinergic agonists. Finally, the PKC activator phorbol myristate acetate had effects opposite to Gö 6976, in that it markedly increased the rate of decrement to the basal [Ca2+]i level after P2Y2 stimulation and also increased the activity of SERCA 2b. These results suggest that SERCA 2b plays a role in regulating the sustained phase of Ca2+ transients caused by P2Y2 stimulation.
- Published
- 2006
30. Multiple anti-inflammatory and anti-atherosclerotic properties of red wine polyphenolic extracts: differential role of hydroxycinnamic acids, flavonols and stilbenes on endothelial inflammatory gene expression
- Author
-
Marika Massaro, Mariangela Pellegrino, Egeria Scoditti, Giovanna Giovinazzo, Nadia Calabriso, Carlo Storelli, Maria Annunziata Carluccio, and Ilaria Ingrosso
- Subjects
0301 basic medicine ,Lipopolysaccharide ,Coumaric Acids ,Flavonols ,Intercellular Adhesion Molecule-1 ,Anti-Inflammatory Agents ,Medicine (miscellaneous) ,Vascular Cell Adhesion Molecule-1 ,Wine ,Resveratrol ,Caftaric acid ,03 medical and health sciences ,chemistry.chemical_compound ,E-selectin ,Stilbenes ,Cell Adhesion ,Humans ,RNA, Messenger ,Chemokine CCL2 ,Inflammation ,Nutrition and Dietetics ,biology ,Cell adhesion molecule ,NF-kappa B ,food and beverages ,Endothelial Cells ,Polyphenols ,Atherosclerosis ,Transcription Factor AP-1 ,Oxidative Stress ,030104 developmental biology ,Biochemistry ,chemistry ,Italy ,biology.protein ,Myricetin ,Kaempferol ,E-Selectin - Abstract
The aim of the study was to evaluate the vascular anti-inflammatory effects of polyphenolic extracts from two typical South Italy red wines, the specific contribution of individual polyphenols and the underlying mechanisms of action. Human endothelial cells were incubated with increasing concentrations (1–50 μg/mL) of Primitivo and Negroamaro polyphenolic extracts (PWPE and NWPE, respectively) or pure polyphenols (1–25 μmol/L), including hydroxycinnamic acids (p-coumaric, caffeic and caftaric acids), flavonols (kaempferol, quercetin, myricetin) or stilbenes (trans-resveratrol, trans-piceid) before stimulation with lipopolysaccharide. Through multiple assays, we analyzed the endothelial–monocyte adhesion, the endothelial expression of adhesion molecules (ICAM-1, VCAM-1 and E-Selectin), monocyte chemoattractant protein-1 (MCP-1) and macrophage colony-stimulating factor (M-CSF), as well as ROS intracellular levels and the activation of NF-κB and AP-1. Both PWPE and NWPE, already at 1 μg/mL, inhibited monocyte adhesion to stimulated endothelial cells, a key event in triggering vascular inflammation. They down-regulated the expression of adhesion molecules, ICAM-1, VCAM-1, E-Selectin, as well as MCP-1 and M-CSF, at mRNA and protein levels. All polyphenols reduced intracellular ROS, and everything, except caftaric acid, inhibited the endothelial expression of adhesion molecules and MCP-1, although with different potency. Flavonols and resveratrol significantly reduced also the endothelial expression and release of M-CSF. The decrease in endothelial inflammatory gene expression was related to the inhibition of NF-κB and AP-1 activation but not to intracellular oxidative stress. This study showed multiple anti-inflammatory and anti-atherosclerotic properties of red wine polyphenolic extracts and indentified specific bioactive polyphenols which could counteract inflammatory diseases including atherosclerosis.
- Published
- 2014
31. Differential response of normal, dedifferentiated and transformed thyroid cell lines to cisplatin treatment
- Author
-
Francesco Paolo Fanizzi, Bruno Di Jeso, Antonella Ciccarese, Loredana Urso, Nadia Calabriso, Carlo Storelli, Santo Marsigliante, Danilo Migoni, Antonella Muscella, Muscella, Antonella, Urso, L., Calabriso, N., Ciccarese, Antonella, Migoni, D., Fanizzi, Francesco Paolo, DI JESO, Bruno, Storelli, Carlo, and Marsigliante, Santo
- Subjects
MAPK/ERK pathway ,Cisplatin ,ERK ,PC-Cl3 ,PKB/Akt ,PKC-ζ ,Animals ,Antineoplastic Agents ,Blotting, Western ,Cell Differentiation ,Cell Line ,Cell Line, Transformed ,Extracellular Signal-Regulated MAP Kinases ,Phosphatidylinositol 3-Kinases ,Phosphorylation ,Proto-Oncogene Proteins c-akt ,Rats ,Thyroid Gland ,Biochemistry ,chemistry.chemical_compound ,LY294002 ,Protein kinase A ,Protein kinase B ,Protein kinase C ,Pharmacology ,biology ,Blotting ,Kinase ,Molecular biology ,Transformed ,chemistry ,Mitogen-activated protein kinase ,biology.protein ,Western - Abstract
The effects of cisplatin ( cis Pt) on the extra cellular signal-regulated kinase (ERK) and the protein kinase B (PKB/Akt), known to play important roles in promoting cell survival and in down regulating apoptosis, were investigated in thyroid cell lines. The cytotoxic effect of cis Pt was highest in normal PC-Cl3 cells, intermediate in dedifferentiated PC-E1A and PC-raf cells and lowest in fully transformed and tumorigenic PC-E1Araf cells. Cis Pt provoked ERK phosphorylation; such phosphorylation was unaltered by Go6976, a conventional PKC inhibitor, whilst blocked by low doses (0.1 μM) or high doses (10 μM) of GF109203X, an inhibitor of all PKC isozymes, in PC-Cl3 and in PC-E1Araf cells, respectively. In PC-E1Araf, but not in PC-Cl3 cells, the cis Pt-provoked ERK phosphorylation was also blocked by a myristoylated PKC-ζ pseudo substrate peptide (PS-ζ). The cytotoxic effects of cis Pt increased when cells were pre-incubated with the mitogen-activated protein kinase (MEK) inhibitor PD98059. Cis Pt provoked the phosphorylation of PKB/Akt and this effect was blocked by LY294002, a PI3K inhibitor. In PC-Cl3 cells pre-incubated with LY294002 the effects of cis Pt on ERK phosphorylation and cell mortality resulted unaffected; conversely, LY294002 reduced the ERK phosphorylation and increased cis Pt cytotoxity of in PC-E1Araf cells. Furthermore, in PC-E1Araf cells pre-incubated with LY294002 and PS-ζ ERK phosphorylation was abolished and cis Pt cytotoxicity was highest. Altogether results highlight a role for PKCs in the upstream regulation of ERK pathway facing the cell response to cis Pt treatments. Understanding the mechanisms by which cells process cis Pt provides important insights for designing more efficient platinum-based drugs.
- Published
- 2005
32. Formation and characterization of glutamate dehydrogenase monolayers on silicon supports
- Author
-
Laura Blasi, G. Vasapollo, Raffaele Acierno, Carlo Storelli, R. Cingolani, Giuseppe Ciccarella, Michele Maffia, Liberato Manna, Pier Paolo Pompa, Ross Rinaldi, Luigia Longo, Antonia Rizzello, L., Blasi, L., Longo, P. P., Pompa, L., Manna, Ciccarella, Giuseppe, Vasapollo, Giuseppe, Cingolani, Roberto, Rinaldi, Rosaria, A., Rizzello, Acierno, Raffaele, Storelli, Carlo, and Maffia, Michele
- Subjects
Silicon ,Immobilized enzyme ,Biomedical Engineering ,Biophysics ,Fluorescence spectrometry ,Infrared spectroscopy ,Biosensing Techniques ,Microscopy, Atomic Force ,Fluorescence spectroscopy ,Glutamate Dehydrogenase ,Spectroscopy, Fourier Transform Infrared ,Electrochemistry ,Fourier transform infrared spectroscopy ,enzyme monolayer ,Chemistry ,Glutamate dehydrogenase ,technology, industry, and agriculture ,General Medicine ,Enzymes, Immobilized ,NAD ,silicon support ,Biochemistry ,Covalent bond ,Oxidation-Reduction ,Biosensor ,Biotechnology - Abstract
In this paper we have tested two different procedures (the “three-step” and the “four-step” procedures) for the covalent immobilization of glutamate dehydrogenase (GDH) onto silicon supports. Atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), fluorescence spectroscopy and an enzymatic assay were used to probe the structure and activity of the immobilized enzyme. Our results demonstrate that coupling through the “three-step” procedure does not significantly affect either the fold pattern or the activity of the enzyme, suggesting that this method could be ideally suited to the development of high quality monolayers for use in enzyme-based planar biosensors.
- Published
- 2005
33. Effect of Cryopreservation on Sea Bass Sperm Proteins
- Author
-
V. Zonno, Sebastiano Vilella, Roberta Schiavone, Carlo Storelli, Loredana Zilli, Rocco Rossano, Zilli, Loredana, Schiavone, Roberta, Zonno, Vincenzo, Rossano, R, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Fish Proteins ,Male ,Semen ,Xenopus Proteins ,Biology ,Peptide Mapping ,sperm ,Cryopreservation ,Andrology ,Human fertilization ,gamete biology ,medicine ,Animals ,Electrophoresis, Gel, Two-Dimensional ,Sea bass ,Polyacrylamide gel electrophoresis ,Sperm motility ,Genetics ,Cell Biology ,General Medicine ,Zebrafish Proteins ,Spermatozoa ,Sperm ,medicine.anatomical_structure ,Reproductive Medicine ,fertilization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Gamete ,Bass ,Semen Preservation - Abstract
In the present study we used two-dimensional polyacryl amide gel electrophoresis (2-DE) and matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to verify whether the protein expression of sea bass sperm was affected by the cryopreservation procedure. The protein profiles differed between fresh and frozen-thawed semen as revealed by visual inspection and by image analysis software. We identified 163 spots in fresh sperm; among these, 13 were significantly decreased and 8 were absent in two-dimensional gel obtained with cryopreserved sperm. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in bio-informatics analysis (Pept-Ident, Mascot, and MS-Fit). In particular, spot 5 showed homology with a novel protein of zebrafish (similar to SKB1 of human and mouse), spot 13 showed homology with amphibian G1/S-specific cyclin E2, and spot 20 showed homology with the hypothetical protein DKFZp566A1524 of Brachidanio rerio. The present work shows that the use of the cryopreservation procedure causes the degradation of sperm proteins and among these, two could be at least partially responsible for the observed decrease in sperm motility duration and the lower hatching rate of eggs fertilized with cryopreserved sperm.
- Published
- 2005
34. Atypical PKC-ζ and PKC-ι mediate opposing effects on MCF-7 Na+/K+ATPase activity
- Author
-
Antonella Muscella, Santo Marsigliante, Carlo Storelli, Muscella, Antonella, Storelli, Carlo, and Marsigliante, Santo
- Subjects
Cytochalasin D ,Indoles ,Physiology ,Morpholines ,Sodium-Potassium-Exchanging ATPase ,ATPase ,Blotting, Western ,Clinical Biochemistry ,Breast Neoplasms ,Culture Media, Serum-Free ,Oligodeoxyribonucleotides, Antisense ,Maleimides ,Wortmannin ,chemistry.chemical_compound ,Cell Line, Tumor ,Animals ,Humans ,RNA, Messenger ,Enzyme Inhibitors ,Na+/K+-ATPase ,Protein Kinase C ,Protein kinase C ,Cellular localization ,Phosphoinositide-3 Kinase Inhibitors ,MCF-7 breast cancer cell line ,Dose-Response Relationship, Drug ,PKC-iota ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Chemistry ,Angiotensin II ,Na/K ATPase ,Cell Biology ,PKC-zeta ,Molecular biology ,Androstadienes ,Isoenzymes ,Kinetics ,Chromones ,biology.protein ,Cattle ,Female ,Intracellular - Abstract
We demonstrated previously that in serum-starved MCF-7 breast cancer cell line, Ang II increased Na+/K+ATPase activity and activated the protein kinase C zeta (PKC-zeta) (Muscella et al., 2002 J Endocrinol 173:315-323; 2003 J Cell Physiol 197:61-68.). The aim of the present study was to investigate the modulation of the activity of the Na+/K+ATPase by PKC-zeta in MCF-7 cells. Here, using serum-starved MCF-7 cells, we have demonstrated that the effect of Ang II on the Na+/K+ATPase activity was inhibited by a synthetic myristoylated peptide with sequences based on the endogenous PKC-zeta pseudosubstrate region (zeta-PS) and by high doses of GF109203X, inhibitor of PKCs. When MCF-7 cells, grown in 10% fetal bovine serum (FBS), were stimulated with Ang II a dose- and time-dependent inhibition of the Na+/K+ATPase activity was obtained. Under this growth condition we found that mRNAs for AT1, AT2, and for Na+/K+ATPase alpha1 and alpha3 subunits were unchanged; besides both the activity of the Na+/K+ATPase and the level of PKC-zeta also were unaffected by the serum. The atypical PKC-iota level (present in very low abundance in serum-starved MCF-7) was increased and Ang II provoked its translocation from the cytosol to plasma membrane. PKC-zeta was localized to the membrane, and upon Ang II treatment its cellular localization did not change. The Ang II-mediated decrease of the Na+/K+ATPase activity was inhibited by high doses of GF109203X but not by zeta-PS, thus indicating that such effect was not due to PKC-zeta activity. The treatment of cells with PKC-iota antisense oligodeoxynucleotides inhibited the effects of Ang II on the Na+/K+ATPase activity. Additionally, the effect of Ang II on Na+/K+ATPase activity was also blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002, and by the actin depolymerizing agents, cytochalasin D. In conclusion, in MCF-7 cells Ang II modulates the Na+/K+ATPase activity by both atypical PKC-zeta/-iota. The effects of Ang II are opposite depending upon the presence of the serum-sensitive PKC-iota, with the inhibitory effect possibly due to the redistribution of sodium pump from plasma membrane to the inactive intracellular pool.
- Published
- 2005
35. Differential signalling of purinoceptors in HeLa cells through the extracellular signal-regulated kinase and protein kinase C pathways
- Author
-
Carlo Storelli, Simona Greco, Antonella Muscella, Santo Marsigliante, M. G. Elia, Muscella, Antonella, Greco, S, Elia, Mg, Storelli, Carlo, and Marsigliante, Santo
- Subjects
MAPK/ERK pathway ,Physiology ,Clinical Biochemistry ,Mitogen-activated protein kinase kinase ,Biology ,Uridine Diphosphate ,Phosphatidylinositol 3-Kinases ,Cytosol ,Humans ,purinergic ,PKC ,Enzyme Inhibitors ,Protein kinase A ,Protein Kinase C ,Protein kinase C ,Flavonoids ,MAP kinase kinase kinase ,Kinase ,Receptors, Purinergic ,Cell Biology ,Molecular biology ,Cell biology ,Enzyme Activation ,Isoenzymes ,ERK ,HeLa cancer cell ,Phosphorylation ,Calcium ,Mitogen-Activated Protein Kinases ,Sodium-Potassium-Exchanging ATPase ,Signal transduction ,Proto-Oncogene Proteins c-fos ,Cell Division ,HeLa Cells ,Signal Transduction - Abstract
We have previously shown that HeLa cells express P2Y2 and P2Y6 receptors endogenously and determined the pathways by which the P2Y2 controls proliferation and Na+/K+ATPase activity. Our objective in this study was to investigate the hypothesis that P2Y6 also controls proliferation and Na+/K+ATPase activity; the pathways used in these actions were partially characterised. We found that P2Y6 activation controlled cell proliferation but not the activity of the Na+/K+ATPase. UDP activation of P2Y6 provoked: (a) an increase in free cytosolic calcium; (b) the activation of protein kinase C-alpha, -beta, -delta, -epsilon, and -zeta but not of PKC-iota and -eta; (c) the phosphorylation of the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2); (d) the expression of c-Fos protein. The P2Y6 induced cell proliferation was blocked by the mitogen-activated protein kinase kinase (MAPKK) inhibitor PD098059, thereby indicating that the ERK pathway mediates the mitogenic signalling of P2Y6. PKC and phosphoinositide 3-kinase (PI3K) inhibitors were tested at two different time points of ERK1/2 phosphorylation (10 and 60 min). The results suggest that novel PKCs and PI3K initiate the response but both conventional and atypical PKCs are required for the maintenance of the UDP-induced phosphorylation of ERK1/2. The induction of c-Fos was greatly diminished by conventional or atypical PKC-zeta inhibition, suggesting that it may be due to PKC-alpha/beta and -zeta activity. These observations demonstrate that UDP acts as a proliferative agent in HeLa cells activating multiple signalling pathways involving conventional, novel, and atypical PKCs, PI3K, and ERK. Of these pathways, conventional and atypical PKCs appear responsible for the induction of c-Fos, while ERK is responsible for cell proliferation and depends upon both novel and atypical PKCs and PI3K activities.
- Published
- 2004
36. Molecular and functional characterisation of the zebrafish (Danio rerio) PEPT1-type peptide transporter1
- Author
-
Alessandro Romano, Carlo Storelli, Tiziano Verri, Michele Maffia, Natascia Tiso, Gabor Kottra, Mark Peric, Francesco Argenton, Michael Boll, and Hannelore Daniel
- Subjects
chemistry.chemical_classification ,animal structures ,food.ingredient ,biology ,fungi ,Biophysics ,Morphogenesis ,Danio ,Transporter ,Peptide ,Cell Biology ,biology.organism_classification ,Biochemistry ,Molecular biology ,Cell biology ,food ,chemistry ,Structural Biology ,Peptide transport ,Yolk ,embryonic structures ,Genetics ,Extracellular ,Molecular Biology ,Zebrafish - Abstract
We report the molecular and functional characterisation of a novel peptide transporter from zebrafish, orthologue to mammalian and avian PEPT1. Zebrafish PEPT1 is a low-affinity/high-capacity system. However, in contrast to higher vertebrate counterparts in which maximal transport activity is independent of extracellular pH, zebrafish PEPT1 maximal transport rates unexpectedly increase at alkaline extracellular pH. Zebrafish pept1 is highly expressed in the proximal intestine since day 4 post-fertilisation, thus preceding functional maturation of the gut, first feeding and complete yolk resorption. Zebrafish PEPT1 might help to understand the evolutionary and functional relationships among vertebrate peptide transporters. Moreover, zebrafish pept1 can be a useful marker for screening mutations that affect gut regionalisation, differentiation and morphogenesis.
- Published
- 2003
37. Changes in cell type composition and enzymatic activities in the hepatopancreas of Marsupenaeus japonicus during the moulting cycle
- Author
-
Carlo Storelli, Sebastiano Vilella, Tiziano Verri, Loredana Zilli, V. Zonno, Roberta Schiavone, G. Scordella, L., Zilli, R., Schiavone, G., Scordella, V., Zonno, Verri, Tiziano, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Male ,medicine.medical_specialty ,Cell type ,Physiology ,ATPase ,Hepatopancreas ,Cell Count ,Marsupenaeus ,Molting ,Biochemistry ,Moulting cycle ,Islets of Langerhans ,Endocrinology ,Decapoda ,Internal medicine ,medicine ,Animals ,Inverse correlation ,Ecology, Evolution, Behavior and Systematics ,Adenosine Triphosphatases ,chemistry.chemical_classification ,biology ,Hepatopancrea ,Fasting ,biology.organism_classification ,shrimp Marsupenaeus japonicu ,Enzyme ,chemistry ,biology.protein ,Female ,Animal Science and Zoology ,Composition (visual arts) ,Moulting - Abstract
The goal of the present study was to evaluate the changes in the cell type composition and ATPase activities (total ATPase, ouabain-sensitive Na+/K(+)-ATPase, furosemide-sensitive Na(+)-ATPase) that occur during the different stages of the moulting cycle in the hepatopancreas of the Marsupenaeus japonicus. The results clearly suggest that the number of resorptive and fibrillar cell types changes significantly during the different stages. An inverse correlation between resorptive and fibrillar cells is observed during moulting (both in normally fed and fasted animals). Fasting, but not the moulting cycle, affects the number of blister-like cells. In the resorptive cells the enzymatic activities (total ATPases and ouabain-sensitive Na+/K(+)-ATPase) also change during the moulting in a cyclical manner. All these results are in agreement with and confirm the different functions carried out by the two cell types within the hepatopancreas.
- Published
- 2003
38. Olive Oil and Red Wine Antioxidant Polyphenols Inhibit Endothelial Activation
- Author
-
Maria Annunziata Carluccio, Maria Assunta Ancora, Carlo Storelli, Luisa Siculella, Francesco Visioli, Alessandro Distante, Marika Massaro, Raffaele De Caterina, Egeria Scoditti, Carluccio, Ma, Siculella, Luisa, Ancora, Ma, Massaro, M, Scoditti, E, Storelli, Carlo, Visioli, F, Distante, A, and DE CATERINA, R.
- Subjects
Transcription, Genetic ,Arteriosclerosis ,Iridoid Glucosides ,Vascular Cell Adhesion Molecule-1 ,Wine ,Biology ,Resveratrol ,Antioxidants ,chemistry.chemical_compound ,Phenols ,Oleuropein ,Stilbenes ,Cell Adhesion ,Animals ,Humans ,Plant Oils ,Iridoids ,RNA, Messenger ,VCAM-1 ,Cell adhesion ,Olive Oil ,Cells, Cultured ,Elenolic acid ,Pyrans ,Flavonoids ,Cell adhesion molecule ,NF-kappa B ,Polyphenols ,U937 Cells ,Phenylethyl Alcohol ,Diet ,Transcription Factor AP-1 ,Tyrosol ,Gene Expression Regulation ,chemistry ,Biochemistry ,Hydroxytyrosol ,Cattle ,Endothelium, Vascular ,Cardiology and Cardiovascular Medicine - Abstract
Objective—Epidemiology suggests that Mediterranean diets are associated with reduced risk of cardiovascular disease. Because monocyte adhesion to the endothelium is crucial in early atherogenesis, we evaluated whether typical olive oil and red wine polyphenols affect endothelial–leukocyte adhesion molecule expression and monocyte adhesion.Methods and Results—Phytochemicals in olive oil and red wine, including oleuropein, hydroxytyrosol, tyrosol, elenolic acid, and resveratrol, with or without antioxidant activity, were incubated with human umbilical vein endothelial cells for 30 minutes, followed by co-incubation with bacterial lipopolysaccharide or cytokines to trigger adhesion molecule expression. At nutritionally relevant concentrations, only oleuropein, hydroxytyrosol, and resveratrol, possessing a marked antioxidant activity, reduced monocytoid cell adhesion to stimulated endothelium, as well as vascular cell adhesion molecule-1 (VCAM-1) mRNA and protein by Northern analysis and cell surface enzyme immunoassay. Reporter gene assays with deletional VCAM-1 promoter constructs indicated the relevance of nuclear factor-κB, activator protein-1, and possibly GATA binding sites in mediating VCAM-1 transcriptional inhibition. The involvement of nuclear factor-κB and activator protein-1 was finally demonstrated at electrophoretic mobility shift assays.Conclusions—Olive oil and red wine antioxidant polyphenols at nutritionally relevant concentrations transcriptionally inhibit endothelial adhesion molecule expression, thus partially explaining atheroprotection from Mediterranean diets.
- Published
- 2003
39. Activation of P2Y2 purinoceptor inhibits the activity of the Na+/K+-ATPase in HeLa cells
- Author
-
Simona Greco, Santo Marsigliante, M. G. Elia, Carlo Storelli, Antonella Muscella, Muscella, Antonella, Elia, Mg, Greco, S, Storelli, Carlo, and Marsigliante, Santo
- Subjects
Gene isoform ,chemistry.chemical_element ,Calcium ,Calcium in biology ,Receptors, Purinergic P2Y2 ,HeLa ,Adenosine Triphosphate ,Humans ,heterocyclic compounds ,RNA, Messenger ,Na+/K+-ATPase ,Receptor ,Protein Kinase C ,Messenger RNA ,Dose-Response Relationship, Drug ,Phospholipase C ,biology ,Receptors, Purinergic P2 ,Chemistry ,Cell Biology ,biology.organism_classification ,Molecular biology ,Kinetics ,Sodium-Potassium-Exchanging ATPase ,HeLa Cells ,Signal Transduction - Abstract
The role of ATP on regulation of the Na(+)/K(+)-ATPase activity in the human cancerous HeLa cells was investigated. HeLa cells stimulated with increasing ATP concentrations showed a dose-dependent inhibition of the Na(+)/K(+)-ATPase activity. These effects were also obtained by UTP. ATP and UTP provoked a rise in intracellular calcium concentration ([Ca(2+)](i)) persisting for at least 4 min. The inhibitor of phospholipase C, U73122, blocked the elevation of [Ca(2+)](i) provoked by ATP/UTP. The expression of mRNA for P2Y2 and P2Y6 receptors was demonstrated by RT-PCR. ATP/UTP activated PKC-alpha, -betaI and -epsilon isoforms, but not PKC-delta and -zeta. The inhibition of the Na(+)/K(+)-ATPase activity by ATP/UTP was blocked by Gö6976, a specific inhibitor of the calcium-dependent PKCs. In conclusion, our results suggest that ATP/UTP modulate Na(+)/K(+)-ATPase activity in HeLa cells through the P2Y2 purinoceptor via calcium mobilisation and activation of calcium-dependent PKCs.
- Published
- 2003
40. Characterization of Na+/H+ Antiporter Activity in PC-Cl3 Thyroid Cells
- Author
-
Santo Marsigliante, Sebastiano Vilella, Loredana Zilli, Carlo Storelli, and Roberta Schiavone
- Subjects
chemistry.chemical_classification ,Gene isoform ,Messenger RNA ,medicine.diagnostic_test ,Physiology ,Antiporter ,Intracellular pH ,Iodide ,Molecular biology ,Amiloride ,Biochemistry ,chemistry ,Western blot ,medicine ,Intracellular ,medicine.drug - Abstract
Background/Aims: In thyroid cells, the intracellular pH plays a key role in the control of the iodide uptake, since iodide accumulation is associated with an intracellular acidification. In the present paper we studied the kinetic proprieties of the Na+/H+ antiporter (NHE) and the molecular expression of different NHE isoforms in rat thyroid PC-Cl3 cells. In addition the intracellular buffer capacity was also evaluated. Methods: pHi was measured using the pH sensitive fluorescent dye BCECF-AM. Amiloride, 5-(N,N-Dimethyl) hydrochloride was used to inhibit NHE activity. RT-PCR and western blot analyses were used to study the expression of NHE mRNA and protein isoforms. Results: PC-Cl3 cells shown a resting pHi, in the absence of CO2/HCO3-, of 6.94 ± 0.1; after an acid load PC-Cl3 cells recovered toward resting pHi value, using a Na-dependent H+ extrusion mechanisms which was amiloride sensitive (Ki = 23 µM). The kinetic parameters were K(Na)app = 10 ± 2 mM and Vmax = 0.23 ± 0.02 ΔpH/min x 105 cells. NHE1, NHE2 and NHE3 were expressed at the mRNA level as well as at the protein level. Conclusion: PC-Cl3 cells express a functional Na/H exchange activity and different isoforms (NHE1, NHE2 and NHE3) are expressed in the plasma membrane.
- Published
- 2003
41. Activation of angiotensin II type I receptor promotes protein kinase C translocation and cell proliferation in human cultured breast epithelial cells
- Author
-
Antonella Muscella, Simona Greco, M. G. Elia, Paola Salvatore, Santo Marsigliante, Carlo Storelli, S., Greco, Muscella, Antonella, Elia, M. G., Salvatore, P., Storelli, Carlo, and Marsigliante, Santo
- Subjects
medicine.medical_specialty ,Angiotensin receptor ,Indoles ,Thapsigargin ,Endocrinology, Diabetes and Metabolism ,Carbazoles ,Losartan ,Receptor, Angiotensin, Type 1 ,Angiotensin Receptor Antagonists ,chemistry.chemical_compound ,Endocrinology ,Internal medicine ,medicine ,Humans ,Staurosporine ,Breast ,Calcium Signaling ,Enzyme Inhibitors ,Estrenes ,Protein Kinase C ,Protein kinase C ,Analysis of Variance ,Receptors, Angiotensin ,Angiotensin II receptor type 1 ,Phospholipase C ,Chemistry ,Angiotensin II ,Biological Transport ,Epithelial Cells ,Pyrrolidinones ,Enzyme Activation ,Isoenzymes ,Type C Phospholipases ,cardiovascular system ,Female ,Cell Division ,hormones, hormone substitutes, and hormone antagonists ,medicine.drug - Abstract
The effect of angiotensin II (Ang II) on Ca(2+) signalling in human primary cultured breast epithelial cells was investigated by using fura-2 as the Ca(2+) probe. Ang II (0.1-1000 nM) induced an intracellular free calcium ([Ca(2+)](i)) transient peak which was unchanged by external Ca(2+ )removal. In Ca(2+)-free medium pretreatment with thapsigargin abolished Ang II-induced Ca(2+ )release. Suppression of 1,4,5-inositol trisphosphate formation by U73122, a phospholipase C inhibitor, blocked the Ang II-induced Ca(2+) response. Losartan (DuP753), an inhibitor of Ang II type I receptor (AT1), decreased the [Ca(2+)](i) increase evoked by Ang II, while CGP4221A, an inhibitor of Ang II type II receptor (AT2) did not. AT1 desensitisation was demonstrated with respect to the Ca(2+) response after subsequent exposure of cells to Ang II and also after pretreatment for 25 min with 1000 nM phorbol 12-myristate 13-acetate. Staurosporine, an inhibitor of protein kinases C (PKC), inhibited the AT1 desensitisation. Epithelial breast cells expressed PKC-alpha, -beta1, -delta and -zeta isozymes, and Ang II provoked translocation from the cytosol to the membranes of PKC-alpha, -beta1, and -delta (but not -zeta). Ang II was also able to stimulate cell proliferation in a dose-dependent manner; this effect was blocked by Go 6976, a specific inhibitor of PKC-alpha and -beta1, the Ca(2+)-dependent isozymes. The main conclusion of this study is that the the Ang II signalling mechanism in breast epithelial cells is based on the elevation of [Ca(2+)](i )released from intracellular stores through AT1 activation. In addition, Ang II stimulates cell proliferation by the activation of PKC isozymes.
- Published
- 2002
42. Muscarinic acetylcholine receptor activation induces Ca2+ mobilization and Na+/K+-ATPase activity inhibition in eel enterocytes
- Author
-
E Jiménez, Simona Greco, Santo Marsigliante, Antonella Muscella, M. G. Elia, Carlo Storelli, Muscella, Antonella, Greco, S., Elia, M. G., Jimenez, E., Storelli, Carlo, and Marsigliante, Santo
- Subjects
endocrine system ,medicine.medical_specialty ,Thapsigargin ,Nifedipine ,Endocrinology, Diabetes and Metabolism ,Calcineurin Inhibitors ,Calcium-Transporting ATPases ,Cholinergic Agonists ,Naphthalenes ,Tacrolimus ,Calcium in biology ,chemistry.chemical_compound ,Endocrinology ,Internal medicine ,medicine ,Animals ,Receptors, Cholinergic ,Calphostin ,Channel blocker ,Enzyme Inhibitors ,Protein Kinase C ,Sirolimus ,Analysis of Variance ,Eels ,Dose-Response Relationship, Drug ,Phospholipase C ,Calcium channel ,Sodium ,Calcium Channel Blockers ,Enzyme Activation ,EGTA ,Enterocytes ,Calphostin C ,chemistry ,Type C Phospholipases ,Calcium ,Carbachol ,Sodium-Potassium-Exchanging ATPase - Abstract
The effect of carbachol (Cch) on intracellular calcium concentration ([Ca2+]i) in eel enterocytes was examined using the fluorescent Ca2+ indicator fura-2. Cch caused a biphasic increase in [Ca2+]i, with an initial spike followed by a progressively decreasing level (over 6 min) to the initial, pre-stimulated, level. The effect of Cch was dose-dependent with a 7.5-fold increase in [Ca2+]i over basal level induced by the maximal dose of Cch (100 microM). In Ca2+-free/EGTA buffer the effect of Cch was less pronounced and the [Ca2+]i returned rapidly to basal levels. The increment of [Ca2+]i was dose-dependently attenuated in cells pre-treated with U73122, a specific inhibitor of phospholipase C, suggesting that the Cch-stimulated increment of [Ca2+]i required inositol triphosphate formation. In the presence of extracellular Ca2+, thapsigargin (TG), a specific microsomal Ca2+-ATPase inhibitor, caused a sustained rise in [Ca2+]i whereas in Ca2+-free medium the increase in [Ca2+]i was transient; in both cases, subsequent addition of Cch was without effect. When 2 mM CaCl2 were added to the cells stimulated with TG or with Cch in Ca2+-free medium, a rapid increase in [Ca2+]i was detected, corresponding to the capacitative Ca2+ entry. Thus, both TG and Cch depleted intracellular Ca2+ stores and stimulated influx of extracellular Ca2+ consistent with capacitative Ca2+ entry. K+ depolarization obtained with increasing concentrations of KCl in the extracellular medium induced a dose-related increase in [Ca2+]i which was blocked by 2 microM nifedipine, a non-specific L-type Ca2+ channel blocker. Nifedipine also changed significantly the height of the Ca2+ transient, and the rate of decrement to the pre-stimulated [Ca2+]i level, indicating that Ca2+ entry into enterocytes also occurs through an L-type voltage-dependent calcium channel pathway. We also show that isolated enterocytes stimulated with increasing Cch concentrations (0.1-1000 microM) showed a dose-dependent inhibition of the Na+/K+-ATPase activity. The threshold decrease was at 1 microM Cch; it reached a maximum at 100 microM (50.5% inhibition) and did not decrease further with the use of higher dose. The effect of Cch on Na+/K+-ATPase activity was dependent on both protein kinase C (PKC) and protein phosphatase calcineurin activation since the PKC inhibitor calphostin C abolished Cch effects, while the calcineurin inhibitor FK506 augmented Cch effect. Collectively, these data establish a functional pathway by which Cch can modulate the activity of the Na+/K+-ATPase through a PKC-dependent (calphostin C-sensitive) pathway and a calcineurin-dependent (FK506-sensitive) pathway.
- Published
- 2002
43. Carbonic anhydrase activity in tissues of the icefish Chionodraco hamatus and of the red-blooded teleosts Trematomus bernacchii and Anguilla anguilla
- Author
-
Mariella Rollo, Carlo Storelli, Michele Maffia, R Chiloiro, Antonia Rizzello, Raffaele Acierno, Maffia, Michele, Rizzello, Antonia, Acierno, Raffaele, M., Rollo, R., Chiloiro, Storelli, Carlo, Rizzello, A, Acierno, R, Rollo, M, and Chiloiro, R
- Subjects
Gills ,Gill ,Gene isoform ,animal structures ,Nothotheniodei ,Physiology ,carbonic anhydrase ,Antarctic Regions ,Antarctic teleost ,Trematomus bernacchii ,Aquatic Science ,Kidney ,Hemoglobins ,blood ,Chionodraco hamatus ,Carbonic anhydrase ,Trematomus ,medicine ,Animals ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Carbonic Anhydrases ,Chaenichthyidae ,chemistry.chemical_classification ,Anguilla anguilla ,biology ,Fishes ,Carbonic anhydrase activity ,gill ,Anatomy ,Anguilla ,biology.organism_classification ,pH homeostasi ,Intestines ,medicine.anatomical_structure ,Enzyme ,Biochemistry ,chemistry ,haemoglobinle ,Chionodraco hamatu ,Insect Science ,biology.protein ,Animal Science and Zoology - Abstract
SUMMARY Carbonic anhydrase (CA) activity was measured in blood, intestine, kidney and gill of two Antarctic teleosts, the haemoglobinless Chionodraco hamatus and the red-blooded Trematomus bernacchii, and of the temperate teleost Anguilla anguilla. In all species, the highest CA activity was in the gills, with the greatest activity in C. hamatus. CA activity in the blood was highest in A. anguilla, but none was detected in the blood of C. hamatus despite the presence of plasma CA inhibitors. The enzyme was present but its activity was low in the intestine and kidney of all three species. The existence of very high CA activity in C. hamatus gills compared with the red-blooded species was investigated further by isolating and characterising the branchial cytosolic CA isoforms. The turnover rate of the C. hamatus isoform was significantly higher than that of T. bernacchii and A. anguilla. The isoforms from both the Antarctic species exhibited lower apparent Km (Km,app) and heat stability than those from A. anguilla. Sensitivity to sulphonamides was similar in all species and was within the range of the mammalian CA II isoform. The branchial CA isoforms of C. hamatus, T. bernacchii and A. anguilla displayed relative molecular masses of 28.9, 29.9 and 31.2 kDa, respectively. The results suggest that the hemoglobinless teleost possesses a different branchial cytosolic CA isoform from that of red-blooded teleosts.
- Published
- 2001
44. Glucocorticoid receptors in the euryhaline teleost Anguilla anguilla
- Author
-
Santo Marsigliante, Stewart Barker, Carlo Storelli, Eugenio Jiménez, Marsigliante, Santo, Barker, S, Jimenez, E, and Storelli, Carlo
- Subjects
Gills ,Gill ,Gene isoform ,Hydrocortisone ,Isoelectric focusing ,Fresh Water ,Anatomy ,Euryhaline ,Biology ,Anguilla ,Adaptation, Physiological ,Biochemistry ,Kinetics ,Cytosol ,Receptors, Glucocorticoid ,Endocrinology ,Glucocorticoid receptor ,Pi ,Osmoregulation ,Animals ,Protein Isoforms ,Seawater ,Molecular Biology - Abstract
To determine the importance of glucocorticoids in the salt water adaptation of European yellow eel we have evaluated the concentration, affinity and physical properties of glucocorticoid receptors (GR) in gill from both sea water- (SW) and freshwater-adapted (FW) animals. Using ligand binding techniques we demonstrated that high affinity GR were present in both cytosolic and nuclear fractions obtained from whole gill. Isoelectric focusing (IEF) of branchial GR indicated the presence of two distinct species, with pI values of 6.1 and 6.7. The form at pI 6.7 sedimented with a Svedberg constant of 4S on glycerol density gradients while the pI 6.1 sedimented in fractions corresponding to 9S. Treatment of the pI 6.1 form with urea (4 M) resulted in generation of the form with pI 6.7. The evidence thus suggested that the oligomeric urea-sensitive form (pI 6.1) contained a form of GR which, as a monomer, focused at pI 6.7. IEF revealed the same concentrations of the pI 6.7 form in both SW and FW. However, there was significantly more (3-fold) pI 6.1 isoform in FW than in SW, and this form decreased gradually during the course of seawater transfer. A transient increase of the nuclear-bound GR was also observed during SW adaptation. The balance between these forms could represent a dynamic parameter with important implications regarding GR function and gill responses to cortisol in salt water adaptation in teleosts.
- Published
- 2000
45. Ionic regulation in Antarctic teleosts
- Author
-
Daniela Pellegrino, Mariella Rollo, Antonia Rizzello, Raffaele Acierno, Bruno Tota, Carlo Storelli, Michele Maffia, Maffia, Michele, Acierno, Raffaele, M., Rollo, Rizzello, Antonia, Storelli, Carlo, D., Pellegrino, and B., Tota
- Subjects
chemistry.chemical_classification ,biology ,Brush border ,Fatty acid ,biology.organism_classification ,Biochemistry ,chemistry ,Intestinal mucosa ,Chionodraco hamatus ,Carbonic anhydrase ,biology.protein ,Animal Science and Zoology ,Cotransporter ,Unsaturated fatty acid ,Homeostasis - Abstract
This work reports recent data on mechanisms of cold adaptation exhibited by the Antarctic teleosts Trematomus bernacchii and Chionodraco hamatus. Analysis of fatty acid in intestinal mucosa brush border suggested that an increase in unsaturated fatty acid could be a mechanism for the preservation of cell membrane integrity and functionality. The investigation of several transporters involved in the regulation of cell homeostasis (Na+/K+‐AT‐Pase, Na+‐D‐glucose cotransport, Na+/H+ exchanger and Ca++‐AT‐Pase) showed kinetic characteristics that could explain part of the adaptation of these proteins to work at low temperature. The activity of carbonic anhydrase, involved in pH control both at intra‐cellular and systemic level, was related to plasma buffer capacity.
- Published
- 2000
46. Effects of zinc on L-[3H]proline uptake by lobster (Homarus americanus) hepatopancreatic brush-border membrane vesicles
- Author
-
Carlo Storelli, Mahealani K. Monteilh-Zoller, Vincenzo Zonno, and G. A. Ahearn
- Subjects
chemistry.chemical_classification ,Homarus ,biology ,Brush border ,Physiology ,Vesicle ,Proline binding ,chemistry.chemical_element ,Zinc ,Aquatic Science ,biology.organism_classification ,Divalent ,Crystallography ,Membrane ,chemistry ,Insect Science ,Animal Science and Zoology ,Proline ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Nuclear chemistry - Abstract
Epithelial brush-border membrane vesicles (BBMVs) from the hepatopancreas of the lobster Homarus americanus were prepared using a magnesium precipitation technique and employed in transport experiments designed to demonstrate the effects of external and internal divalent cationic heavy metals on the uptake of L-[3H]proline. When BBMVs were exposed to a high external concentration (2.5 mmol l−1) of Cd2+, Cu2+, Fe2+, Mn2+ or Zn2+, L-[3H]proline (0.5 mmol l−1) uptake was significantly (P0.05) on apparent L-[3H]proline binding to the membranes (Km=1.66±0.23 mmol l−1) (means ± S.E.M., N=3). In the presence of 0.5 mmol l−1 l-pipecolate, bilateral zinc-stimulated, carrier-mediated, L-[3H]proline influx was abolished. At low external concentrations of zinc alone (e.g. below 1.0 mmol l−1), L-[3H]proline influx was enhanced by the metal. Enhanced amino acid uptake in the presence of external zinc alone was abolished by L-pipecolate. A model accounting for external and internal zinc enhancements of L-[3H]proline influx by the Na+-dependent L-pipecolate-sensitive IMINO transport system in these membranes is proposed.
- Published
- 1999
47. Angiotensin II stimulates the exchanger in human umbilical vein endothelial cells via AT1 receptor
- Author
-
Antonella Muscella, Eugenio Jiménez, Carlo Storelli, Santo Marsigliante, Sebastiano Vilella, Muscella, Antonella, Marsigliante, Santo, Vilella, Sebastiano, Jimenez, E, and Storelli, Carlo
- Subjects
Umbilical Veins ,medicine.medical_specialty ,Cell Membrane Permeability ,Sodium-Hydrogen Exchangers ,ATPase ,Stimulation ,Receptor, Angiotensin, Type 2 ,Hemostatics ,Receptor, Angiotensin, Type 1 ,General Biochemistry, Genetics and Molecular Biology ,Umbilical vein ,Internal medicine ,medicine ,Humans ,Vasoconstrictor Agents ,General Pharmacology, Toxicology and Pharmaceutics ,Transcellular ,Receptor ,Cells, Cultured ,Fluorescent Dyes ,Receptors, Angiotensin ,Angiotensin II receptor type 1 ,Ionophores ,biology ,Chemistry ,Angiotensin II ,Thrombin ,General Medicine ,Hydrogen-Ion Concentration ,Fluoresceins ,Sodium–hydrogen antiporter ,Spectrometry, Fluorescence ,Endocrinology ,Nigericin ,cardiovascular system ,biology.protein ,Endothelium, Vascular ,Sodium-Potassium-Exchanging ATPase ,hormones, hormone substitutes, and hormone antagonists - Abstract
Angiotensin II (Ang II) has an important role in cardiovascular regulation and in the control of electrolyte balance, and its role in the regulation of Na+ transcellular movements through its actions on the activity of Na + K + ATPase is well documented. We showed previously that human umbilical vein endothelial cells (HUVEC) express the Ang II type 1 (AT1) receptor, which mediates Ang II modulation of Na + K + ATPase activity (1). We here investigate the effects of Ang II on the activity of the Na + H + exchanger in HUVEC. When compared with controls, incubation of HUVEC for 20 min with different concentrations of Ang II provoked significant increases in Na + H + activity. The stimulation was dose dependent between 1 and 10 nM Ang II and varied with time of incubation up to 20 min. The maximal response, obtained with 10 nM Ang II after 20 min treatment, resulted in a 65% increment in Na + H + activity. Preincubation of HUVEC with 10 μM DuP753 blocked Na + H + activation by Ang II. These results suggest that the effects of Ang II on both the Na + K + ATPase and the Na + H + exchanger may increase the transendothelial flux of Na+ and are mediated by the AT1 receptor.
- Published
- 1999
48. Biphasic Scatchard plots of oestrogen receptors are associated with low pS2 levels in human breast cancers
- Author
-
Carlo Storelli, Luciana Biscozzo, Giuseppe Leo, Santo Marsigliante, Marsigliante, Santo, Biscozzo, L, Leo, G, and Storelli, Carlo
- Subjects
Adult ,Cancer Research ,medicine.medical_specialty ,medicine.drug_class ,Mammary gland ,Population ,Breast Neoplasms ,Biology ,Internal medicine ,Gene expression ,medicine ,Humans ,skin and connective tissue diseases ,Receptor ,education ,Aged ,Aged, 80 and over ,chemistry.chemical_classification ,Immunoradiometric assay ,education.field_of_study ,Tumor Suppressor Proteins ,Proteins ,Radioimmunoassay ,Middle Aged ,Enzyme ,medicine.anatomical_structure ,Endocrinology ,Receptors, Estrogen ,Oncology ,chemistry ,Estrogen ,Female ,Trefoil Factor-1 ,Receptors, Progesterone ,hormones, hormone substitutes, and hormone antagonists - Abstract
In a series of 100 breast rumours, oestrogen receptors (ER) were analysed by Scatchard plots, progesterone receptors (PR) by enzyme immunoassay and pS2 by an immunoradiometric assay. Scatchard analysis gave information on receptor heterogeneity in that there was a large variation in K-d values obtained, from 0.001-2.95 nM. This variation was largely confined to tumours containing less than 70 fmol receptor per mg protein, while tumours with higher receptor concentrations were a more homogeneous population with low K-d values. An obvious correlation between ER and PR was found; moreover, pS2 was correlated to both ER and PR. In addition, 20 of the 100 tumours gave biphasic Scatchard plots, indicating the presence of at least two oestrogen-binding moieties, with K-d values and concentrations both in the range of those of receptors, The tumours displaying biphasic Scatchard plots had very low pS2 expression, regardless of ER concentrations; this was not true for PR. It is suggested that variability in responses to endocrine therapy may be related to the heterogeneity of the ER present in breast tumours.
- Published
- 1999
49. Computerised counting of tumour infiltrating lymphocytes in 90 breast cancer specimens
- Author
-
A. Marra, Carlo Storelli, Luciana Biscozzo, Giuseppe Nicolardi, G.B. Lobreglio, Giuseppe Leo, and Santo Marsigliante
- Subjects
Adult ,Cancer Research ,Pathology ,medicine.medical_specialty ,CD3 Complex ,CD8 Antigens ,CD3 ,Mammary gland ,Breast Neoplasms ,chemical and pharmacologic phenomena ,Lymphocytes, Tumor-Infiltrating ,Breast cancer ,Image Interpretation, Computer-Assisted ,Animals ,Humans ,Medicine ,IL-2 receptor ,Lymph node ,Aged ,biology ,Tumor-infiltrating lymphocytes ,business.industry ,Age Factors ,hemic and immune systems ,Histology ,Middle Aged ,medicine.disease ,Immunohistochemistry ,medicine.anatomical_structure ,Oncology ,CD4 Antigens ,biology.protein ,Female ,business - Abstract
Tumour infiltrating lymphocytes (TILs) implicated in immunologic cytotoxicity were evaluated by immunohistochemistry and digitally counted in serial sections from 90 breast cancers in order to assess their number, the relationships between them and to tumour histology. CD3 + , CD4 + , CD8 + , CD20 + , CD25 + and CD56 + lymphocytes were found in 58 (64.4%), 52 (57.7%), 50 (55.5%), 22 (24.4%), 11 (12.2%) and 21 (23.3%) tumours, respectively. There was no difference in the number of TILs between pure infiltrating ductal (NOS) and non-ductal carcinomas, and no relationship between TILs and histological grades was found. CD3 + TILs directly correlated to age, while lymph node negative patients had tumours infiltrated by fewer CD4 + TILs with respect to lymph node positive patients. In 25/90 patients, randomly chosen, the status of peripheral blood lymphocytes was evaluated but no differences with respect to the status found in healthy blood donors was obtained; nonetheless while in some patients CD8 + TILs outnumbered CD4 + TILs in situ, the CD4/CD8 ratio was normal in their peripheral blood. The results show a considerable diversity of TILs among breast tumours, their lack of relationship with the status of the peripheral blood cells, and their potential important relationship with age (CD3 + ) and lymph node status (CD4 + ).
- Published
- 1999
50. Electroneutral Na+/H+exchange in brush-border membrane vesicles fromPenaeus japonicushepatopancreas
- Author
-
V. Zonno, L. Ingrosso, Sebastiano Vilella, Tiziano Verri, and Carlo Storelli
- Subjects
Sodium-Hydrogen Exchangers ,Brush border ,Physiology ,Intracellular pH ,Antiporter ,Sodium ,chemistry.chemical_element ,Lithium ,Amiloride ,chemistry.chemical_compound ,Chlorides ,Penaeidae ,Physiology (medical) ,Electrochemistry ,Extracellular ,Animals ,Ion transporter ,Chromatography ,Microvilli ,Vesicle ,Acridine orange ,Hydrogen-Ion Concentration ,Kinetics ,chemistry ,Calcium ,Digestive System ,Hydrogen ,Nuclear chemistry - Abstract
An electroneutral Na+/H+exchange mechanism (dimethylamiloride inhibitable, Li+sensitive, and Ca2+insensitive) was identified in brush-border membrane vesicles (BBMV) from Kuruma prawn hepatopancreas by monitoring Na+-dependent H+fluxes with the pH-sensitive dye acridine orange and measuring22Na+uptake. Kinetic parameters measured under short-circuited conditions were the Na+concentration that yielded one-half of the maximal dissipation rate ( Fmax) of the preset transmembrane ΔpH ( KNa) = 15 ± 2 mM and Fmax= 3,626 ± 197 Δ F ⋅ min−1⋅ mg protein−1, with a Hill coefficient for Na+of ∼1. In addition, the inhibitory constant for dimethylamiloride was found to be ∼1 μM. The electroneutral nature of the antiporter was assessed in that an inside-negative transmembrane electrical potential neither affected kinetic parameters nor stimulated pH-dependent (intracellular pH > extracellular pH)22Na+uptake. In contrast, electrogenic pH-dependent22Na+uptake was observed in lobster hepatopancreatic BBMV. Substitution of chloride with gluconate resulted in increasing KNaand decreasing Δ Fmax, which suggests a possible role of chloride in the operational mechanism of the antiporter. These results indicate that a Na+/H+exchanger, resembling the electroneutral Na+/H+antiporter model, is present in hepatopancreatic BBMV from the Kuruma prawn Penaeus japonicus.
- Published
- 1998
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.