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Summary The Atomic Force Microscope (AFM) is able to image the surface prope1ties of ali types of mate ri a ls (conductors, in sul ators, bio logical and vegetai samples) with a very hi gh resolution and in the native state. ln parti cul... more
Summary The Atomic Force Microscope (AFM) is able to image the surface prope1ties of ali types of mate ri a ls (conductors, in sul ators, bio logical and vegetai samples) with a very hi gh resolution and in the native state. ln parti cul ar, AFM is used to study a number of biologie system such as DNA-replication and pro tein/ prote in , DNA/protein interactions; furthermore, in cosmetology, it's suitabl e to study the fun damental characteri sti cs of the skin , like elasticity and wrinkledness, a nd to compare for exampl e morphological properti es of human virgin hair and c he mical or polymer treated hair. The most important characteri stics of thi s instrume nt in cosmetological analyses are: • the samples can be directe ly examined without previus treatment, i.e. in native conditions, • data are col lect in a di gitai form, • 3-D image rendering with vari able magnification and s hading, • view of the recorded image from a ny point of the space. Jn our laboratory we an...
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Breast cancer is the second leading cause of cancer-related death in women in the world, and its management includes a combination of surgery, radiation therapy, chemotherapy, and immunotherapy, whose effectiveness depends largely, but... more
Breast cancer is the second leading cause of cancer-related death in women in the world, and its management includes a combination of surgery, radiation therapy, chemotherapy, and immunotherapy, whose effectiveness depends largely, but not exclusively, on the molecular subtype (Luminal A, Luminal B, HER2+ and Triple Negative). All breast cancer subtypes are accompanied by peculiar and substantial changes in sphingolipid metabolism. Alterations in sphingolipid metabolite levels, such as ceramides, dihydroceramide, sphingosine, sphingosine-1-phosphate, and sphingomyelin, as well as in their biosynthetic and catabolic enzymatic pathways, have emerged as molecular mechanisms by which breast cancer cells grow, respond to or escape therapeutic interventions and could take on diagnostic and prognostic value. In this review, we summarize the current landscape around two main themes: 1. sphingolipid metabolites, enzymes and transport proteins that have been found dysregulated in human breast...
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Additional file 4. Bibliography for Additional files 1, 2, and 3.
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Additional file 3. Measurement of nitrite concentrations in BMDM treated with synthetic hemozoin.
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Additional file 2. Measurement of urea levels in BMDM treated with malarial hemozoin.
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Additional file 1. Measurement of nitrite and nitrate concentrations in BMDM treated with malarial hemozoin.
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INTRODUCTION: Leishmania tropica is the etiological agent of cutaneous leishmaniasis. Pattern recognition receptors such as toll-like receptors or nucleotide oligomerization domain-like receptors (NLR) have been associated with the... more
INTRODUCTION: Leishmania tropica is the etiological agent of cutaneous leishmaniasis. Pattern recognition receptors such as toll-like receptors or nucleotide oligomerization domain-like receptors (NLR) have been associated with the disease. The role in innate immune responses against L. tropica of NOD2, a member of the NLR family, was investigated. MATERIALS AND METHODS: Interferon gamma (IFN-\u3b3)-primed or unprimed immortalized mouse bone marrow macrophages (BMDM)-wild type (WT) or -NOD2 -/- were infected with L. tropica at different parasite/macrophage ratios (i.e. 2.5, 5, or 10:1), for 24h. Controls, such as medium alone, muramyldipeptide (MDP), or lipopolysaccharide (LPS), were included. Levels of cytokines or nitrite released into supernatants were measured through ELISA or Griess reagents. Levels of inducible nitric oxide synthase (iNOS) mRNA and protein were retrieved through Real-Time PCR and Western blot analyses, respectively. Also, unprimed BMDM- RIP2 -/- or -CARD9 -/- , two downstream components of NOD2 activation, were stimulated with the same ratios of L. tropica or controls, and the production of the pro-inflammatory cytokine TNF-a was compared with BMDM-WT. RESULTS AND CONCLUSIONS: Data showed that L. tropica did not induce the production of nitric oxide in unprimed BMDM-WT. L. tropica induced higher levels of nitric oxide in IFN-\u3b3- primed BMDM-WT than in unstimulated cells. Stimulation due to L. tropica, or the control MDP (known to activate NOD2), was abrogated in the BMDM-NOD2 -/- , but not LPS, as so the expression levels of inducible nitric oxide synthase (iNOS) mRNA or protein. In addition, NOD2, RIP2 or CARD9 showed to be involved in the induction of TNF-a release from BMDM. These data suggest an involvement of NOD2 pathway in innate immune response to L. tropica infection
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Summary In the epid ermis of terrestrial vertebrates stratum corneum lipids are known to pl ay an important ro le as regulators of skin permeabi lity; particu larly ceramides, some of whi c h with very long chain, hydroxy fatty acids,... more
Summary In the epid ermis of terrestrial vertebrates stratum corneum lipids are known to pl ay an important ro le as regulators of skin permeabi lity; particu larly ceramides, some of whi c h with very long chain, hydroxy fatty acids, are important components of this "barri er" function. Ceramides are derived from the hydrolys is of previous existing glucosylceramide during the terminal stages of epidermal differentiation, due to the presence of a li sosomal-like glu cosylceramid ase locali sed in the lamellar bodies in the space between stratum granulosum and the inferior layer of stratum corneum: the defi ciency of glu cosylceramidase, local ized in lysosome, is responsible of Gaucher's disease. In thi s study we investigateci the glycolipi ds and ceramide di stribution in th e epiderm is of 4 Gau cher's patients (two of type I and two of type 3), compaired wi th th e same parameters in 4 healthy donors. The comparison between the pathological sampl es and the...
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Research Interests: Biology, Medicine, Gene expression, Cell Division, Biological Sciences, and 15 moreDexamethasone, Cell line, Cell Differentiation, Humans, Prolactin, Physical sciences, Phosphorylation, Epidermal Growth Factor, Western blot, Base Sequence, Gangliosides, Dimerization, Growth factor receptor, Tyrosine Phosphorylation, and Medical and Health Sciences
In recent years, there has been a growing interest in natural antioxidants as replacements of synthetic compounds because of increased safety concerns and worldwide trend toward the usage of natural additives in foods. One of the richest... more
In recent years, there has been a growing interest in natural antioxidants as replacements of synthetic compounds because of increased safety concerns and worldwide trend toward the usage of natural additives in foods. One of the richest sources of natural antioxidants, nowadays largely studied for their potential to decrease the risk of diseases and to improve oxidative stability of food products, are edible brown seaweeds. Nevertheless, their antioxidant mechanisms are slightly evaluated and discussed. The aims of this study were to suggest possible mechanism(s) of Fucus vesiculosus antioxidant action and to assess its bioactivity during the production of enriched rye snacks. Chemical and cell-based assays indicate that the efficient preventive antioxidant action of Fucus vesiculosus extracts is likely due to not only the high polyphenol content, but also their good Fe2+-chelating ability. Moreover, the data collected during the production of Fucus vesiculosus-enriched rye snacks ...
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Wound healing is a very complex process that allows organisms to survive injuries. It is strictly regulated by a number of biochemical and physical factors, mechanical forces included. Studying wound healing in space is interesting for... more
Wound healing is a very complex process that allows organisms to survive injuries. It is strictly regulated by a number of biochemical and physical factors, mechanical forces included. Studying wound healing in space is interesting for two main reasons: (i) defining tools, procedures, and protocols to manage serious wounds and burns eventually occurring in future long-lasting space exploration missions, without the possibility of timely medical evacuation to Earth; (ii) understanding the role of gravity and mechanical factors in the healing process and scarring, thus contributing to unravelling the mechanisms underlying the switching between perfect regeneration and imperfect repair with scarring. In the study presented here, a new in vivo sutured wound healing model in the leech (Hirudo medicinalis) has been used to evaluate the effect of unloading conditions on the healing process and the effectiveness of platelet rich plasma (PRP) as a countermeasure. The results reveal that micr...
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Ischemic-reperfusion (I/R) injury induced a remodeling of protein and lipid homeostasis, under oxidative stress and inflammatory status. Starvation occurring during I/R is a condition leading to autophagy activation, which allows abnormal... more
Ischemic-reperfusion (I/R) injury induced a remodeling of protein and lipid homeostasis, under oxidative stress and inflammatory status. Starvation occurring during I/R is a condition leading to autophagy activation, which allows abnormal material clearance or amino acid, or both, and fatty acid (FA) recycling essential for survival. This study investigated the lipid reshaping, peroxidation, and related-signaling pathways, in rat brain endothelial cells (RBE4) subjected to 3 h of oxygen and glucose deprivation (OGD) and restoration of standard condition (I/R in vitro model). Lipids and proteins were analyzed after 1 or 24 h of oxygen and nutrient restoration. Together with the oxidative stress and inflammatory status, I/R injury induced a reshaping of neutral lipids and biogenesis of lipid droplets (LD) with excessive lipid storage. The increase of LC3-II/LC3-I ratio, an autophagy marker, and LC3 co-localization with LD suggest the activation of lipophagy machinery to counteract the...
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Research Interests: Western blotting, Biological Sciences, Cell line, Mice, Animals, and 13 morePhysical sciences, HPTL (High Performance Thin Layer Chromatography), Immunoprecipitation, Phosphotyrosine, Epidermal Growth Factor, Epidermal Growth Factor Receptor, Western blot, Cell Surface Markers, Epithelial Cell Biology, Dimerization, Receptor Tyrosine Kinase, Tyrosine Phosphorylation, and Medical and Health Sciences
Antioxidants are suggested to act as radioprotectors, and dietary supplements based on antiox-idants have been proposed for astronauts involved in long-term space missions. Plant extracts with antioxidant properties may be used in... more
Antioxidants are suggested to act as radioprotectors, and dietary supplements based on antiox-idants have been proposed for astronauts involved in long-term space missions. Plant extracts with antioxidant properties may be used in dietetic supplements for astronauts; in fact recent nutritional guidelines suggest that "fruits and vegetables may become as important on space-going vessels as limes were on the sea-going vessels
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Space flights cause a number of patho-physiological changes. Oxidative damage has been demonstrated in astronauts after space flights. Oxidative stress is due to an imbalance between production of oxidant and antioxidative defence. In... more
Space flights cause a number of patho-physiological changes. Oxidative damage has been demonstrated in astronauts after space flights. Oxidative stress is due to an imbalance between production of oxidant and antioxidative defence. In embryos of Xenopus laevis, the glutathione system is an inducible antioxidant defence. For this reason, we investigated the effect of gravity deprivation on endogenous antioxidant enzymes in X. laevis embryos developed for 6 days in a Random Positioning Machine. The results show that glutathione content and the activity of antioxidant enzymes increase in RPM embryos, suggesting the presence of a protective mechanism. An induction of antioxidant defence might play an important role for animals to adapt to micro-gravitational stress, possibly during actual space flights.
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Cultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short... more
Cultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short culture lifetime, and variations between passages. To standardize thestudies on keratinocytes, we investigated the use of HaCaT cells, a long-lived, spontaneously immortalized human keratinocyte line which is able to differentiate, as a suitable model to follow the release of inflammatory and repair mediators in response to TNFor IL-1. Different treatment conditions (presence or absence of serum) and differentiation stimuli (increase in cell density as a function of time in culture and elevation of extracellular calcium) were considered. ELISA and Multiplex measurement technologies were used to monitor the production of cytokines and chemokines. Taken together, the results highlight that Caconcentration in the medium, cell density, and presence of serum...
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The efficacy of cosmetic products and active substances have been investigated on human keratinocytes cell line on a pool of tests. The work aimed to study the possible modifications of the biological parameters tested (cytotoxicity and... more
The efficacy of cosmetic products and active substances have been investigated on human keratinocytes cell line on a pool of tests. The work aimed to study the possible modifications of the biological parameters tested (cytotoxicity and cytoskeleton morphology) related to the cellular adhesion function. The results have permitted to define different activities, for the different products, compared to the untreated culture and to their placebo.
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ABSTRACT To test the effects of space radiation on living organisms we participated (23 June 2002) to the "BI.R.BA." mission (organised and granted by the Italian Space Agency, A.S.I.), a flight with a stratospheric... more
ABSTRACT To test the effects of space radiation on living organisms we participated (23 June 2002) to the "BI.R.BA." mission (organised and granted by the Italian Space Agency, A.S.I.), a flight with a stratospheric balloon, from Trapani-MILO (ASI Launch site) to Seville in Spain. Xenopus laevis emryos at different stages of development were exposed during flight. Results on mortality show that younger embryos (16 hr) flown on the balloon (B) are more sensitive to radiation exposure compared with older ones (40 hr and 6 days). The exposure during flight lowers the antioxidant potential in all the embryos, particularly in older ones.
All living organisms are constantly exposed to oxidant agents deriving from both endogenous and exogenous sources capable to modify biomolecules and induce damages. Free radicals generated by oxidative stress exert an important role in... more
All living organisms are constantly exposed to oxidant agents deriving from both endogenous and exogenous sources capable to modify biomolecules and induce damages. Free radicals generated by oxidative stress exert an important role in the development of tissue damage and aging. Reactive species (RS) derived from oxygen (ROS) and nitrogen (RNS) pertain to free radicals family and are constituted by various forms of activated oxygen or nitrogen. RS are continuosly produced during normal physiological events but can be removed by antioxidant defence mechanism: the imbalance between RS and antioxidant defence mechanism leads to modifications in cellular membrane or intracellular molecules. In this chapter only endogenous antioxidant molecules will be critically discussed, such as Glutathione, Alpha-lipoic acid, Coenzyme Q, Ferritin, Uric acid, Bilirubin, Metallothioneine, L-carnitine and Melatonin.
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Research Interests: Breast Cancer, Phospholipids, Cell Cycle, Apoptosis, Gene expression, and 17 moreCell line, Humans, Cancer Therapy, Omega-3 fatty acids, Female, Cell Death, Disease Prevention, Cell Viability, Phosphorylation, Caspase, Epidermal Growth Factor, Fatty Acid, Cell Proliferation, Cell Survival, Plasma Membrane, Eicosapentaenoic Acid, and Breast Cancer Cells
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Erythrocyte and hemoglobin losses have been frequently observed in humans during space missions; these observations have been designated as "space anemia". Erythrocytes exposed to microgravity have a modified rheology and... more
Erythrocyte and hemoglobin losses have been frequently observed in humans during space missions; these observations have been designated as "space anemia". Erythrocytes exposed to microgravity have a modified rheology and undergo hemolysis to a greater extent. Cell membrane composition plays an important role in determining erythrocyte resistance to mechanical stress and it is well known that membrane composition might be influenced by external events, such as hypothermia, hypoxia or gravitational strength variations. Moreover, an altered cell membrane composition, in particular in fatty acids, can cause a greater sensitivity to peroxidative stress, with increase in membrane fragility. Solar radiation or low wavelength electromagnetic radiations (such as gamma rays) from the Earth or the space environment can split water to generate the hydroxyl radical, very reactive at the site of its formation, which can initiate chain reactions leading to lipid peroxidation. These reac...
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All human GM3 synthase mRNA variants until now identified predict a protein of 362 amino acids having substrate activity highly restricted to lactosylceramide. In this report we describe the identification of a new GM3 synthase transcript... more
All human GM3 synthase mRNA variants until now identified predict a protein of 362 amino acids having substrate activity highly restricted to lactosylceramide. In this report we describe the identification of a new GM3 synthase transcript containing an additional translation start codon, located upstream and in-frame with that up to now considered unique translation initiation site in the human GM3 synthase gene. In vitro expression studies showed that the new transcript produces a longer form of human GM3 synthase, that is efficiently translocated into the microsomal lumen and glycosylated. Moreover, stable cDNA transfection into mammalian cells gives rise to a threefold increase of GM3 synthase activity, associated to a broader substrate specificity. Although this transcript has been initially identified in the human placenta, RT-PCR analyses verified the expression of an identical mRNA also in undifferentiated HL60 cells, but not in the monocytic lineage. Altogether, these results are the first demonstration of the existence of a new isoform of human GM3 synthase, which could play an important role during HL60 cell differentiation. The functional relevance of the existence of two isoforms of GM3 synthase is also discussed.
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Gangliosides are known to modulate the activation of receptor tyrosine-kinases (RTKs). Recently, we demonstrated the functional relationship between ErbB2 and ganglioside GM(3) in HC11 epithelial cell line. In the present study we... more
Gangliosides are known to modulate the activation of receptor tyrosine-kinases (RTKs). Recently, we demonstrated the functional relationship between ErbB2 and ganglioside GM(3) in HC11 epithelial cell line. In the present study we investigated, in the same cells, the ErbB2 activation state and its tendency to form stable molecular complexes with the epidermal growth factor receptor (EGFR) and with ganglioside GM(3) upon EGF stimulation. Results from co-immunoprecipitation experiments and western blot analyses indicate that tyrosine-phosphorylated ErbB2 and EGFR monomers and stable ErbB2/EGFR high molecular complexes (heterodimers) are formed following EGF stimulation, even if the receptors co-immunoprecipitates also in the absence of the ligand; these data suggest the existence of pre-dimerization inactive receptor clusters on the cell surface. High performance-thin layer chromatography (HP-TLC) and TLC-immunostaining analyses of the ganglioside fractions extracted from the immunoprecipitates demonstrate that GM(3), but not other gangliosides, is tightly associated to the tyrosine-phosphorylated receptors. Furthermore, we show that GM(3) is preferentially and in a SDS-resistant manner associated to the activated ErbB2/EGFR complexes and EGFR monomer, but not to ErbB2. Altogether our data support the hypothesis that the modulating effects produced by GM(3) on ErbB2 activation are mediated by EGFR.
Research Interests: Western blotting, Biological Sciences, Cell line, Mice, Animals, and 10 morePhysical sciences, HPTL (High Performance Thin Layer Chromatography), Immunoprecipitation, Phosphotyrosine, Epidermal Growth Factor, Epidermal Growth Factor Receptor, Western blot, Dimerization, Receptor Tyrosine Kinase, and Tyrosine Phosphorylation
To test the effects of space radiation on living organisms we participated (23 June 2002) to the... more
To test the effects of space radiation on living organisms we participated (23 June 2002) to the "BI.R.BA." mission (organised and granted by the Italian Space Agency, A.S.I.), a flight with a stratospheric balloon, from Trapani-MILO (ASI Launch site) to Seville in Spain. Xenopus laevis emryos at different stages of development were exposed during flight. Results on mortality show that
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Antioxidants are suggested to act as radioprotectors, and dietary supplements based on antiox-idants have been proposed for astronauts involved in long-term space missions. Plant extracts with antioxidant properties may be used in... more
Antioxidants are suggested to act as radioprotectors, and dietary supplements based on antiox-idants have been proposed for astronauts involved in long-term space missions. Plant extracts with antioxidant properties may be used in dietetic supplements for astronauts; in fact recent nutritional guidelines suggest that "fruits and vegetables may become as important on space-going vessels as limes were on the sea-going vessels