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1.
Sci Bull (Beijing) ; 62(14): 974-975, 2017 Jul 30.
Article in English | MEDLINE | ID: mdl-36659497
3.
J Dent Res ; 83(11): 837-42, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15505232

ABSTRACT

The cytotoxicity of dental monomers has been widely investigated, but the underlying mechanisms have not been elucidated. We studied the molecular mechanisms involved in cell death induced by HEMA. In human primary fibroblasts, HEMA induced a dose-dependent apoptosis that was confirmed by the activation of caspases-8, -9, and -3. We found an increase of reactive oxygen species (ROS) and NF-kappaB activation after HEMA exposure. Blocking of ROS production by anti-oxidants had no direct influence on apoptosis caused by HEMA, but inhibition of NF-kappaB increased the fraction of apoptotic cells. Accordingly, mouse embryonic fibroblasts (MEF) from p65-/- mice were more susceptible to HEMA-induced apoptosis than were wild-type controls. Our results indicate that exposure to HEMA triggers apoptosis and that this mechanism is not directly dependent upon redox signaling. Nevertheless, ROS induction by HEMA activates NF-kappaB, which exerts a protective role in counteracting apoptosis.


Subject(s)
Apoptosis/drug effects , Fibroblasts/drug effects , Methacrylates/toxicity , NF-kappa B/physiology , Analysis of Variance , Animals , Blotting, Western , Caspases/metabolism , Cells, Cultured , Electrophoresis, Polyacrylamide Gel , Enzyme Activation , Fibroblasts/cytology , Humans , Mice , Reactive Oxygen Species/metabolism , Skin/cytology
4.
Curr Biol ; 11(8): 614-9, 2001 Apr 17.
Article in English | MEDLINE | ID: mdl-11369207

ABSTRACT

Ras p21 signaling is involved in multiple aspects of growth, differentiation, and stress response [1-2]. There is evidence pointing to superoxides as relays of Ras signaling messages. Chemicals with antioxidant activity suppress Ras-induced DNA synthesis. The inhibition of Ras significantly reduces the production of superoxides by the NADPH-oxidase complex [3]. Kirsten and Harvey are nonallelic Ras cellular genes that share a high degree of structural and functional homology. The sequences of Ki- and Ha-Ras proteins are almost identical. They diverge only in the 20-amino acid hypervariable domain at the COOH termini. To date, their functions remain indistinguishable [4]. We show that Ki- and Ha-Ras genes differently regulate the redox state of the cell. Ha-Ras-expressing cells produce high levels of reactive oxygen species (ROS) by inducing the NADPH-oxidase system. Ki-Ras, on the other hand, stimulates the scavenging of ROS by activating posttranscriptionally the mitochondrial antioxidant enzyme, Mn-superoxide dismutase (Mn-SOD), via an ERK1/2-dependent pathway. Glutamic acid substitution of the four lysine residues in the polybasic stretch at the COOH terminus of Ki-Ras completely abolishes the activation of Mn-SOD, although it does not inhibit ERK1/2-induced transcription. In contrast, an alanine substitution of the cysteine of the CAAX box has very little effect on Mn-SOD activity but eliminates ERK1/2- dependent transcription.


Subject(s)
Genes, ras/physiology , Signal Transduction/physiology , 3T3 Cells , Animals , COS Cells , Cell Line , Chlorocebus aethiops , Mice , Mitogen-Activated Protein Kinase 1/antagonists & inhibitors , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3 , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinases/metabolism , Oxidation-Reduction , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Proto-Oncogene Proteins p21(ras)/genetics , Proto-Oncogene Proteins p21(ras)/metabolism , Rats , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism
5.
Am J Physiol Heart Circ Physiol ; 278(6): H2003-7, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10843899

ABSTRACT

We tested the hypothesis that constriction of cerebral arterioles during acute increases in blood pressure is attenuated by activation of potassium (K(+)) channels. We tested the effects of inhibitors of calcium-dependent K(+) channels [iberiotoxin (50 nM) and tetraethylammonium (TEA, 1 mM)] on changes in arteriolar diameter during acute hypertension. Diameter of cerebral arterioles (baseline diameter = 46 +/- 2 microm, mean +/- SE) was measured using a cranial window in anesthetized rats. Arterial pressure was increased from a control value of 96 +/- 1 mmHg to 130, 150, 170, and 200 mmHg by intravenous infusion of phenylephrine. Increases in arterial pressure from baseline to 130 and 150 mmHg decreased the diameter of cerebral arterioles by 5-10%. Greater increases in arterial pressure produced large increases in arteriolar diameter (i.e., "breakthrough of autoregulation"). Iberiotoxin or TEA inhibited increases in arteriolar diameter when arterial pressure was increased to 170 and 200 mmHg. The change in arteriolar diameter at 200 mmHg was 20 +/- 3% and -1 +/- 4% in the absence and presence of iberiotoxin, respectively. These findings suggest that calcium-dependent K(+) channels attenuate cerebral microvascular constriction during acute increases in arterial pressure, and that increases in arteriolar diameter at high levels of arterial pressure are not simply a passive phenomenon.


Subject(s)
Cerebrovascular Circulation , Homeostasis , Hypertension/physiopathology , Potassium Channels/physiology , 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology , 4-Aminopyridine/pharmacology , Acute Disease , Animals , Arterioles/physiopathology , Blood Pressure/drug effects , Peptides/pharmacology , Potassium Channel Blockers , Rats , Rats, Sprague-Dawley , Reference Values , Tetraethylammonium/pharmacology , Vasoconstriction/drug effects , Vasoconstrictor Agents/pharmacology
6.
Article in English | WPRIM (Western Pacific) | ID: wpr-963791

ABSTRACT

(1) A short outline of the history and development of cerebral angiography in relation to other radiodiagnostic procedures is given(2) Its main indications and contra-indications are pointed out. Given preference in suspected cerebro-vascular pathology (cerebral aneurysms, arterio-venous fistulas, vascular tumors and malformations, etc.), it serves as a supplementary method to pneumographic studies in cerebral tumors and other conditions, where it often gives additional information concerning localization, pathologic type of tumor, and blood supply(3) Cerebral angiographic anatomy and physiology are briefly recalled(4) Radiopaque materials are discussed. Disdract, in the same concentration used for routine renal studies, is preferred. The procedure is considered safe(5) The technique of carotid angiography is escribed. The "open" method of injection into the common carotid is used. X-Ray requirements for a complete cerebral angiography demand a series of at least five radiographics taken within eight seconds; a highly satisfactory home-devised and home-made cassette-changer for these rapid exposures is presented(6) Three cases of cerebral angiography are presentedThis preliminary report on cerebral angiography is made, primarily to arouse the interest of the local medical profession in this, to us, new radiodiagnostic procedure. It is a very interesting field, and we believe that there are many cases in the Islands in which the method would be very informative and diagnostic. Its development will certainly go hand in hand with the development of neuro-surgery; but the general practitioner must be made aware of its possibilities, if he is to keep abreast with medical and surgical progress.(Summary)

7.
Stroke ; 28(11): 2266-71; discussion 2271-2, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9368575

ABSTRACT

BACKGROUND AND PURPOSE: Intracranial blood vessels appear to be relatively resistant to development of atherosclerosis. The goal of this study was to compare effects of mildly oxidized LDL (ox-LDL) on endothelium-dependent responses of intracranial and extracranial arteries in vitro. METHODS: LDL was purified from plasma of healthy subjects and mildly oxidized by means of a xanthine/xanthine oxidase reaction. Contraction of the rabbit basilar and carotid arteries in response to histamine and phenylephrine and relaxation in precontracted vessels to acetylcholine and sodium nitroprusside were evaluated after 30 minutes of exposure to LDL or ox-LDL (100 micrograms/mL). RESULTS: Exposure to LDL had little or no effect on vascular responses. After exposure to ox-LDL, contraction to histamine and phenylephrine and relaxation to acetylcholine were impaired significantly in carotid (P < .05) but not in basilar artery. Relaxation to sodium nitroprusside was not significantly impaired by ox-LDL in the basilar artery. In the carotid artery, relaxation to sodium nitroprusside was significantly impaired by ox-LDL when the vessels were precontracted with phenylephrine but not histamine. Impairment of vascular responses by ox-LDL was prevented by addition of superoxide dismutase, catalase, or dimethylthiourea to the LDL solution before addition of xanthine/xanthine oxidase. CONCLUSIONS: Mildly ox-LDL impairs contraction and endothelium-dependent relaxation in the carotid but not in basilar artery. Thus, intracranial arteries may be relatively resistant to mildly ox-LDL.


Subject(s)
Basilar Artery/drug effects , Carotid Arteries/drug effects , Lipoproteins, LDL/pharmacology , Animals , Basilar Artery/physiology , Carotid Arteries/physiology , Endothelium, Vascular/physiology , Histamine/pharmacology , Humans , Male , Phenylephrine/pharmacology , Potassium Chloride/pharmacology , Rabbits , Vasoconstrictor Agents/pharmacology , Vasodilation/drug effects , Vasodilation/physiology
8.
Am J Physiol ; 272(3 Pt 2): H1287-91, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9087603

ABSTRACT

We examined the hypothesis that activity of Ca2+-dependent K+ channels is increased in the basilar artery during chronic hypertension. Diameter of the basilar artery was measured using a cranial window in anesthetized normotensive Wistar-Kyoto rats (WKY, arterial pressure = 109 +/- 3 mmHg, mean +/- SE) and stroke-prone spontaneously hypertensive rats (SHRSP, arterial pressure = 179 +/- 4 mmHg). Responses of the basilar artery to topical application of tetraethylammonium ion (TEA), an inhibitor of Ca2+-dependent K+ channels, were examined in WKY and SHRSP. Vessel diameter decreased by 2 +/- 1 and 4 +/- 0.1% in WKY and by 7 +/- 2 and 18 +/- 1% in SHRSP (P < 0.05 vs. WKY) in response to 10(-4) and 10(-3) M TEA, respectively. Similar results were obtained using iberiotoxin (10(-8) and 10(-7) M), a highly selective inhibitor of Ca2+-dependent K+ channels. In contrast to constrictor responses to TEA and iberiotoxin, constrictor responses of the basilar artery in response to serotonin and U-46619 were similar in WKY and SHRSP. In WKY rats that were made chronically hypertensive (arterial pressure = 172 +/- 6 mmHg) after treatment for 4 wk with N(G)-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase, constriction of the basilar artery in response to TEA was also enhanced. These findings suggest that activity of Ca2+-dependent K+ channels is enhanced in the basilar artery in vivo in two models of chronic hypertension. Thus Ca2+-dependent K+ channels in the basilar artery may be activated during chronic hypertension, perhaps as a response to elevation of intracellular concentration of Ca2+.


Subject(s)
Basilar Artery/physiopathology , Calcium/pharmacology , Hypertension/physiopathology , Muscle Contraction , Potassium Channels/physiology , Vasoconstriction/drug effects , 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid , Animals , Basilar Artery/physiology , Muscle Contraction/drug effects , Peptides/pharmacology , Potassium Channel Blockers , Prostaglandin Endoperoxides, Synthetic/pharmacology , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Serotonin/pharmacology , Tetraethylammonium , Tetraethylammonium Compounds/pharmacology , Thromboxane A2/analogs & derivatives , Thromboxane A2/pharmacology , Vasoconstrictor Agents/pharmacology , Wasp Venoms/pharmacology
9.
Stroke ; 28(1): 181-5, 1997 Jan.
Article in English | MEDLINE | ID: mdl-8996509

ABSTRACT

BACKGROUND AND PURPOSE: Adrenomedullin is a recently discovered vasoactive peptide that is structurally related to calcitonin gene-related peptide (CGRP). Adrenomedullin is produced by vascular endothelium and smooth muscle and is present in the brain. The goals of this study were to determine (1) whether adrenomedullin produces dilatation of cerebral arterioles and whether this effect is mediated by activation of CGRP receptors and (2) whether vasodilatation to adrenomedullin was mediated by K+ channels. METHODS: Diameter of cerebral arterioles (mean +/- SE baseline, 46 +/- 1 microns) was measured using a closed cranial window in anesthetized rats. RESULTS: Application of rat adrenomedullin (10(-7) and 10(-6) mol/L) increased vessel diameter by 16 +/- 3% and 45 +/- 8% (n = 5), respectively. Vasodilator responses to repeated application of adrenomedullin were reproducible. Pretreatment of cerebral arterioles with the specific CGRP1 receptor antagonist CGRP-(8-37) (5 x 10(-7) mol/L) selectively inhibited the vasodilator responses to adrenomedullin without inhibiting responses to ADP (10(-5) to 10(-3) mol/L). Responses to adrenomedullin (10(-7) and 10(-6) mol/L) were 14 +/- 1% and 40 +/- 3% before and 2 +/- 2% and 6 +/- 1% after CGRP-(8-37), respectively (P < .01). Glibenclamide (10(-6) mol/L), an inhibitor of ATP-sensitive K+ channels, reduced the responses to adrenomedullin without attenuating responses to ADP. Responses to adrenomedullin were 19 +/- 4% and 35 +/- 6% before and 6 +/- 3% and 19 +/- 5% after glibenclamide, respectively (P < .05). Iberiotoxin (10(-7) mol/L), an inhibitor of calcium-dependent K+ channels, also significantly attenuated responses to adrenomedullin and did not inhibit vasodilatation to papaverine. Responses to adrenomedullin were 16 +/- 2% and 55 +/- 8% before and 12 +/- 4% and 26 +/- 3% after iberiotoxin, respectively (P < .01 for 10(-6) mol/L adrenomedullin). CONCLUSIONS: Adrenomedullin produces substantial dilatation of cerebral arterioles in vivo, and the response is mediated in large part by activation of CGRP1 receptors. Cerebral vasodilatation to adrenomedullin appears to be dependent on activation of K+ channels.


Subject(s)
Arterioles/physiology , Cerebral Arteries/physiology , Peptides/pharmacology , Vasodilation/physiology , Vasodilator Agents/pharmacology , Adenosine Diphosphate/pharmacology , Adrenomedullin , Animals , Arterioles/drug effects , Calcitonin Gene-Related Peptide/pharmacology , Calcitonin Gene-Related Peptide Receptor Antagonists , Cerebral Arteries/drug effects , Glyburide/pharmacology , Male , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/physiology , Peptide Fragments/pharmacology , Potassium Channel Blockers , Rats , Rats, Sprague-Dawley , Vasodilation/drug effects
10.
Stroke ; 27(9): 1603-7; discussion 1607-8, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8784136

ABSTRACT

BACKGROUND AND PURPOSE: The mechanisms by which cAMP and cGMP produce vasorelaxation are not entirely clear. In this study we examined the hypothesis that relaxation of cerebral arterioles in response to receptor-mediated activation of adenylate cyclase (increase in cAMP) is mediated through Ca(2+)-dependent K+ channels. METHODS: We measured the diameter of cerebral arterioles (basal diameter, 47 +/- 1 microns) using an open cranial window in anesthetized rats. Agonists and antagonists were applied locally in the cranial window. RESULTS: Topical application of adenosine (0.1 and 1 mmol/L), a receptor-mediated activator of adenylate cyclase, and dibutyryl cAMP (60 and 200 mumol/L), a cell-permeable analogue of cAMP, dilated cerebral arterioles. Iberiotoxin (50 nmol/L), a selective inhibitor of Ca(2+)-dependent K+ channels, reduced vasodilatation in response to 0.1 and 1 mmol/L adenosine by 66% and 28%, respectively. Tetraethylammonium (TEA) (1 mmol/L), another inhibitor of Ca(2+)-dependent K+ channels, reduced vasodilatation to 0.1 and 1 mmol/L adenosine by 58% and 42%, respectively, and reduced vasodilatation in response to 60 and 200 mumol/L dibutyryl cAMP by 75% and 66%, respectively. Topical application of sodium nitroprusside (0.1 and 1 mumol/L), a direct activator of guanylate cyclase, and 8-bromo cGMP (200 and 600 mumol/L), a cell-permeable analogue, produced dilatation of cerebral arterioles that was inhibited by iberiotoxin (50 nmol/L) and TEA (1 and 3 mmol/L). In contrast, dilatation of cerebral arterioles in response to papaverine (which produces vasodilatation in large part by inhibition of Ca2+ channels) and aprikalim (which produces vasodilation by activation of ATP-sensitive K+ channels) was not inhibited by iberiotoxin or TEA. CONCLUSIONS: These findings suggest that dilatation of cerebral arterioles by receptor-mediated activation of adenylate cyclase and by direct activation of guanylate cyclase in the rat is mediated in large part by activation of Ca(2+)-dependent K+ channels.


Subject(s)
Calcium/physiology , Cerebrovascular Circulation/physiology , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Potassium Channels/physiology , Vasodilation/physiology , Adenosine/pharmacology , Adenylyl Cyclases/metabolism , Animals , Arterioles/drug effects , Arterioles/metabolism , Arterioles/physiology , Cerebrovascular Circulation/drug effects , Enzyme Activation , Guanylate Cyclase/metabolism , Rats , Rats, Sprague-Dawley
11.
Stroke ; 25(12): 2457-60; discussion 2461-2, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7974590

ABSTRACT

BACKGROUND AND PURPOSE: The goal of this study was to determine whether release of endothelium-derived relaxing factor (EDRF) from carotid artery in response to acetylcholine is altered by aging. METHODS: Responses were examined in arteries from Wistar rats 6 to 8 months old (young rats), 24 to 26 months old (old rats), and 30 to 32 months old (very old rats). We used a bioassay technique to measure release of EDRF from the carotid artery (donor vessel) and, in the same experiment, measured the diameter of pressurized donor arteries. RESULTS: Responses of the donor arteries and the detector vessels (aortas) to sodium nitroprusside were not altered in old (24 to 26 months) and very old (30 to 32 months) rats compared with responses in the young rats. Dilator responses in the carotid artery to acetylcholine tended to be lower in the old rats and were significantly lower in very old rats compared with the young rats. Relaxation of the detector vessel during administration of acetycholine to the donor artery was not significantly different in young, old, and very old rats. CONCLUSIONS: Endothelium-dependent dilatation in carotid arteries of old rats is impaired, but release of EDRF and responsiveness of vascular muscle to nitroprusside are normal. Thus, impairment of endothelium-dependent relaxation in the carotid artery during aging is not due to impaired release of EDRF.


Subject(s)
Acetylcholine/pharmacology , Aging/metabolism , Carotid Artery, Common/drug effects , Carotid Artery, Common/metabolism , Nitric Oxide/metabolism , Animals , Aorta, Thoracic/drug effects , Aorta, Thoracic/metabolism , Biological Assay , Blood Pressure , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Image Processing, Computer-Assisted , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/metabolism , Nitroprusside/pharmacology , Phenylephrine/pharmacology , Rats , Rats, Wistar , Vasoconstriction , Vasodilation/drug effects
12.
Neuroreport ; 3(2): 177-80, 1992 Feb.
Article in English | MEDLINE | ID: mdl-1623168

ABSTRACT

Purified synaptic vesicles are highly enriched with a protein which binds cell plasma membranes. The binding is selective for acidic phospholipids and sialoglycosphingolipids. In partition chromatography of vesicle proteins, the binding activity was co-eluted with a limited set of proteins. Among them the most abundant species were two vesicle-specific proteins: p65 and synaptophysin. In affinity chromatography of vesicle proteins, only p65 bound to a column of immobilized lysoganglioside. The same protein, purified by preparative electrophoresis, retained the binding activity and fully reproduced the hemagglutinating property and the selectivity for acidic lipids of whole vesicles. The results suggest that the (hemagglutinating) lipid binding properties of the vesicles are mainly if not exclusively due to p65.


Subject(s)
Calcium-Binding Proteins , Membrane Glycoproteins/metabolism , Nerve Tissue Proteins/metabolism , Phospholipids/metabolism , Sphingolipids/metabolism , Synaptic Vesicles/metabolism , Cell Membrane/metabolism , Chromatography, Affinity , Electrophoresis, Polyacrylamide Gel , Hemagglutination/physiology , Humans , Immunoblotting , Membrane Glycoproteins/isolation & purification , Nerve Tissue Proteins/isolation & purification , Protein Binding , Synaptotagmin I , Synaptotagmins
13.
Acta Neurol (Napoli) ; 13(3): 213-9, 1991 Jun.
Article in English | MEDLINE | ID: mdl-1927629

ABSTRACT

Synaptic vesicles purified from rat forebrain contain a protein with membrane-binding activity. The binding has a high degree of selectivity for sialoglycosphingolipids. The protein was partially purified by affinity chromatography on lysoGM1 silica beads. The protein bands thus isolated have an apparent Mr of 65 and 53 KDa respectively.


Subject(s)
G(M1) Ganglioside/metabolism , Synaptic Vesicles/chemistry , Animals , Protein Binding , Rats , Rats, Inbred Strains
14.
Neuroreport ; 2(2): 93-5, 1991 Feb.
Article in English | MEDLINE | ID: mdl-1883990

ABSTRACT

Synaptic vesicles contain a protein (agglutinin) which binds cell plasma membranes. The protein activity is titrated measuring the agglutination of rabbit trypsinized-fixed red blood cells induced by serial dilutions of purified vesicles. The protein is highly concentrated in vesicles: the specific activity is 150-fold higher in vesicles compared to the brain homogenate, and 20-fold higher compared to the crude synaptosomal membrane fraction. The binding is specifically inhibited by the four major brain sialoglycosphingolipids, with a higher affinity for polysialo- rather than for monosialoganglioside (GM1) IC50 are 3.9 x 10(-5) M (GT1b), 8.9 x 10(-5) M (GD1a), 2.8 x 10(-4) M (GD1b) and 4.9 x 10(-4) M (GM1). Cells which are not agglutinated by the vesicles (human red blood cells) can be activated by incubation with gangliosides: insertion of the glycolipids in the plasma membranes makes them sensitive to the vesicle haemagglutinin.


Subject(s)
Nerve Tissue Proteins/metabolism , Synaptic Vesicles/metabolism , Animals , Cell Membrane/metabolism , Erythrocytes/metabolism , Female , Gangliosides/metabolism , Gangliosides/pharmacology , Hemagglutinins/metabolism , Male , Rats , Rats, Inbred Strains
15.
Int J Cancer ; 34(3): 335-7, 1984 Sep 15.
Article in English | MEDLINE | ID: mdl-6480154

ABSTRACT

Incidence rates of lip cancer in males in Ragusa (Sicily) are amongst the highest in Europe [age-standardized rate (world) for 1980-82: 7.5 per 100,000]. A case-control study was conducted on 53 male cases and 106 controls matched for sex, age (+/- 2 1/2 years), residence and hospital from which cases had been drawn. Individual interviews were carried out for the evaluation of ethnic, environmental, pathologic and occupational risk factors. Lip cancer was significantly associated with: fair, brown, or red hair (relative odds = 2.3), blue eyes (r.o. = 5.3), fair skin (r.o. = 8.0), sensitivity to sunburns (r.o. = 4.1), working outdoors (r.o. = 4.9), coexistence of non-specific lesions of exposed body parts (r.o. = 12,2), low socioeconomic status (r.o. = 15.8), farming (r.o. = 2.6) and working in greenhouses (r.o. = 12.0). Recall of recurrent Herpes labialis was not significantly associated; also association with tobacco smoking was not significant after adjustment for socioeconomic status. Risk increased exponentially with the number of ethnic characteristics of northern European populations.


Subject(s)
Lip Neoplasms/epidemiology , Adult , Age Factors , Aged , Ethnicity , Humans , Italy , Male , Middle Aged , Registries , Risk
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