Phoneutria nigriventer (armed spider) venom induces increased vascular permeability in rat and rabbit skin in vivo

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOThe effect of intradermally injected Phoneutria nigriventer venom (PNV) on vascular permeability of both rat and rabbit skin has been investigated. Oedema formation was measured as the local extravascular accumulation at skin sites of intravenously injected 125I-human serum albumin. In both rat and rabbit PNV induced dose-dependent oedema which was greatly potentiated by the vasodilators calcitonin-gene-related peptide and prostaglandin E1. In rats, PNV-induced oedema was markedly reduced either by previous treatment of the animals with the histamine H1 antagonist mepyramine and the serotonin antagonist methysergide or when venom was dialysed, indicating a major role for histamine and serotonin. In rabbits, dialysis of the venom to remove histamine and serotonin did not reduce PNV-induced oedema, indicating presence of oedematogenic component(s) which are different from the amines histamine or serotonin.The effect of intradermally injected Phoneutria nigriventer venom (PNV) on vascular permeability of both rat and rabbit skin has been investigated. Oedema formation was measured as the local extravascular accumulation at skin sites of intravenously injected 125I-human serum albumin. In both rat and rabbit PNV induced dose-dependent oedema which was greatly potentiated by the vasodilators calcitonin-gene-related peptide and prostaglandin E1. In rats, PNV-induced oedema was markedly reduced either by previous treatment of the animals with the histamine H1 antagonist mepyramine and the serotonin antagonist methysergide or when venom was dialysed, indicating a major role for histamine and serotonin. In rabbits, dialysis of the venom to remove histamine and serotonin did not reduce PNV-induced oedema, indicating presence of oedematogenic component(s) which are different from the amines histamine or serotonin30910111016FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO90/12341; 90/4145-

Phoneutria nigriventer spider venom induces oedema in rat skin by activation of capsaicin sensitive sensory nerves

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOPhoneeutria nigriventer venom induces oedema formation when injected in the rat dorsal skin and such oedema is, in part, dependent on the stimulation of tachykinin NK1 receptors. This study investigated whether Phoneutria nigriventer venom acts directly on tachykinin NK1 receptors, or indirectly to activate sensory neurones which in turn release a tachykinin NK1 receptor agonist. The plasma extravasation induced by Phoneutria nigriventer venom (1-10 mu g/site) in neonatally capsaicin (8-methyl N-vanillyl-6-nonenamide)-pretreated rats was substantially attenuated (P < 0.05) but the response to either the tachykinin NK1 receptor agonist GR73632 ((delta Ava[L-Pro(9), N-MeLeu10] substance P-(7-11) 30 pmol/site) or bradykinin (0.3-3 nmol/site) was not affected. These results indicate that Phoneutria nigriventer venom stimulates sensory nerves indirectly. The lack of effect of capsaicin-pretreatment on the GR73632 and bradykinin responses indicated that the tachykinin NK1 and bradykinin B-2 receptors remained functional. There was no evidence to suggest that Phoneutria nigriventer venom contains a tachykinin NK1 receptor agonist3392-3223226FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOsem informaçã

Phoneutria Nigriventer (armed Spider) Venom Induces Increased Vascular Permeability In Rat And Rabbit Skin In Vivo

The effect of intradermally injected Phoneutria nigriventer venom (PNV) on vascular permeability of both rat and rabbit skin has been investigated. Oedema formation was measured as the local extravascular accumulation at skin sites of intravenously injected 125I-human serum albumin. In both rat and rabbit PNV induced dose-dependent oedema which was greatly potentiated by the vasodilators calcitonin-gene-related peptide and prostaglandin E1. In rats, PNV-induced oedema was markedly reduced either by previous treatment of the animals with the histamine H1 antagonist mepyramine and the serotonin antagonist methysergide or when venom was dialysed, indicating a major role for histamine and serotonin. In rabbits, dialysis of the venom to remove histamine and serotonin did not reduce PNV-induced oedema, indicating presence of oedematogenic component(s) which are different from the amines histamine or serotonin. © 1992.30910111016Antunes, Marangoni, Borges, Fontana, de Nucci, Pharmacological profile of Phoneutria nigriventer venom on rabbit vascular smooth muscle (1990) British Journal of Pharmacology, 101, p. 508PBrain, Williams, Inflammatory oedema induced by synergism between calcitonin gene-related peptide (CGRP) and mediators of increased vascular permeability (1985) Br. J. Pharmac., 86, pp. 855-860Diniz, Cordeiro, Junior, Kelly, Fischer, Reiman, Oliveira, Richardson, The purification and amino acid sequence of the lethal neurotoxin Tx1 from the venom of the Brazilian ‘armed’ spider Phoneutria nigriventer (1990) FEBS Lett., 263, pp. 251-253Entwistle, Johnstone, Medzihradszky, May, Isolation of a pure toxic polypeptide from the venom of the spider Phoneutria nigriventer and its neurophysiological activity on an insect femur preparation (1982) Toxicon, 20, pp. 1059-1067Fontana, Vital-Brazil, Mode of action of Phoneutria nigriventer spider venom at the isolated phrenic nerve-diaphragm of the rat (1985) Braz. J. Med. Biol. Res., 18, pp. 557-565Kaiser, The enzymatic activity of spider venom (1953) Mem. Inst. Butantan, 25, pp. 35-39Lucas, Spiders in Brazil (1988) Toxicon, 26, pp. 759-772Rezende, Jr, Cordeiro, Oliveira, Diniz, Isolation of neurotoxic peptides from the venom of the armed spider Phoneutria nigriventer (1991) Toxicon, 29, pp. 1225-1233Schenberg, Pereira-Lima, Phoneutria nigriventer venom (1971) Pharmacology and biochemistry of its components, 3, pp. 279-297. , W. Bucherl, E.E. Buckley, Venomous Animals and their Venoms, Academic Press, New YorkSpector, Willoughby, Endogenous mediators of increased vascular permeability in inflammation (1968) The Pharmacology of Inflammation, pp. 22-54. , English Universities Press LtdVital-Brazil, Leite, Fontana, Modo de ação da peçonha da aranha armadeira, Phoneutria nigriventer (Keyserling, 1891), nas aurículas isoladas de cobaia (1988) Ciênc. Cult., S Paulo, 40, pp. 181-185Williams, Prostaglandin E2, prostaglandin I2 and the vascular changes of inflammation (1979) Br. J. Pharmac., 65, pp. 517-52

Involvement of vanilloid receptors and purinoceptors in the phoneutria nigriventer spider venom-induced plasma extravasation in rat skin

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOPhoneutria nigriventer venom causes stimulation of capsaicin-sensitive primary afferent neurons in the rat dorsal skin, leading to neurogenic plasma protein extravasation due to the release of tachykinin NK1 receptor agonist. In this study we further investigated the mechanisms involved in the venom-induced activation of capsaicin-sensitive primary afferent neurons. The plasma extravasation in response to venom intradermally injected was measured in Wistar rats as the local accumulation of i.v. injected I-125-labelled human serum albumin into skin sites. The tachykinin NK1 receptor agonist, D-Ala-[L-Pro(9),Me-Leu(8)]substance P-(7-11) (GR73632; 10-100 pmol/site), induced a significant plasma leakage that was abolished by the selective tachykinin NK1 receptor antagonist, (S)-1-[2-[3-(3,4-dichlorphenyl)-1 (3-isopropoxyphenylacetyl) piperidin-3-yl] ethyl]-4-phenyl-1 azaniabicyclo [2.2.2]octane chloride (SR140333; 1 nmol/site), whereas the leakage after venom (1-10 mu g/site) was significantly inhibited (but not abolished) by SR140333. The calcitonin gene-related peptide (CGRP) receptor antagonist, CGRP-(8-37), failed to further reduce the residual plasma extravasation induced by venom plus SR140333, The mu-opioid receptor agonist, [D-Ala(2),Me-Phe(4),Gly-ol(5)]enkephalin (DAMGO), and the local anaesthetic, Lignocaine, had no effect on the venom-induced plasma extravasation. Similarly, the L-, N- and P/Q-type voltage-sensitive Ca2+ channel blockers (verapamil, omega-conotoxin MVIIA and MVIIC, respectively) as well as the Na+ channel blockers, tetrodotoxin and carbamazepine, had no effect on the venom-induced effect. Neither the systemic treatment nor the local injection of ruthenium red prevented the venom-induced plasma extravasation. However, the vanilloid receptor antagonist, N-[2-(4-chlorophenyl) ethyl]-1,3,4,5-tetrahydro7,8-dihydroxy-2H-2-benzazepine-2-carbothioamide (capsazepine; 120 mu mol/kg, i.v.), reduced by 48% (P <0.05) the venom (10 mu g/site)-induced plasma extravasation. A significant inhibitory effect was also observed with the P, purinoceptor agonists, adenosine 5'-triphosphate (ATP; 10 and 30 nmol/site) and adenosine 5'-diphosphate (ADP; 10 nmol/site). The involvement of histamine and/or 5-hydroxytryptamine (5-HT) in the venom-induced plasma extravasation was ruled out since neither histamine and 5-HT receptor antagonists nor depletion of mast cells by compound 48/80 affected the venom response. This was further supported by the failure of venom to degranulate in vitro peritoneal mast cells. In conclusion, only vanilloid receptors and P, prejunctional purinoceptors had an inhibitory effect on the neurogenic plasma extravasation evoked by P. nigriventer venom in rat dorsal skin3913305315FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOsem informaçã

Activation By Phoneutria Nigriventer (armed Spider) Venom Of Tissue Kallikrein-kininogen-kinin System In Rabbit Skin In Vivo

1. The purpose of the present study was to investigate the mechanisms by which venom from Phoneutria nigriventer spider induces increases in vascular permeability in rabbit skin. 2. Local oedema formation, in response to intradermally -injected agents, was measured in male New Zealand white rabbits as the local accumulation of i.v. injected 125I-labelled human serum albumin into skin sites. 3. Phoneutria nigriventer venom (10-30 μg/site) increased vascular permeability, which was inhibited by trasylol (10 μg/site) and the bradykinin B2 receptor antagonists D-Arg, [Hyp3,Thi5,8,D-Phe7]-BK (3 nmol/site) and Hoe 140 (0.3 nmol/site). In addition, the oedema induced by the venom was potentiated by the kinase II inhibitor, captopril (1 nmol/site). The lipoxygenased inhibitor, BWA4C (10 nmol/site) and the PAF antagonist, WEB 2086 (100 nmol/site) had no effect on the venom-induced increase in vascular permeability. 4. Incubation of rabbit plasma with Phoneutria nigriventer venom in vitro did not cause bradykinin formation. Further, the plasma kallikrein inhibitor, soybean trypsin inhibitor (10 μg/site), had no effect on the venom-induced increase in vascular permeability in rabbit skin. 5. These results indicate that the oedema produced by Phoneutria nigriventer venom is dependent on the activation of the tissue kallikrein-kinin system.109253954

Activation Of Tissue Kallikrein-kininogen-kinin System In Rabbit Skin By A Fraction Isolated From Phoneutria Nigriventer (armed Spider) Venom

Phoneutria nigriventer venom was fractionated by gel filtration followed by ion-exchange chromatography from which 16 fractions (I-XVI) were obtained and assayed in rabbit skin in order to identify those responsible for the increased vascular permeability observed with the whole venom. The fractions, and control mediators (tissue kallikrein, bradykinin and histamine) were intradermally injected in male New Zealand white rabbits. Local oedema formation was measured as the local accumulation of i.v. injected 125I-human serum albumin into skin sites. Fraction XIII was the only fraction assayed which significantly induced oedema formation. Fraction XIII-induced oedema was greatly reduced by either the protease inhibitor aprotinin or the bradykinin B2 receptor antagonist d-Arg,[Hyp3,Thi5,8,d-Phe7]-Bk, whereas the plasma kallikrein inhibitor soybean trypsin inhibitor failed to significantly affect this oedematogenic response. The kininase II inhibitor captopril markedly potentiated fraction XIII-induced oedema. Our results indicate that the increased vascular permeability induced by fraction XIII is due to local generation of kinins in response to tissue (but not plasma) kallikrein-kinin system activation. © 1993.311113851391Antunes, Marangoni, Brain, de Nucci, Phoneutria nigriventer (armed spider) venom induces increased vascular permeability in rat and rabbit skin in vivo (1992) Toxicon, 30, pp. 1011-1016Antunes, Marangoni, Borges, Hyslop, Fontana, de Nucci, Effect of Phoneutria nigriventer on rabbit vascular smooth muscle (1993) Braz. J. Med. Biol. Res., 26, pp. 81-91Brain, Williams, Inflammatory oedema induced by synergism between calcitonin gene-related peptide (CGRP) and mediators of increased vascular permeability (1985) Br. J. Pharmac., 86, pp. 855-860Brazil, Vellard, Contribuição ao estudo de venenos de aranhas (1925) Mem. Inst. Butantan, 2, pp. 1-70Brazil, Vellard, Contribuição ao estudo do veneno das aranhas II (1926) Mem. Inst. Butantan, 3, pp. 3-77Brazil, Vellard, Contribuição ao estudo do veneno das aranhas III (1926) Mem. Inst. Butantan, 3, pp. 243-294Chao, Tanaka, Margolius, Inhibitory effects of sodium and other monovalent cations on purified versus membrane-bound kallikrein (1983) J. biol. Chem., 258, pp. 6461-6465Cushman, Cheung, Sabo, Ondetti, Design of potent competitive inhibitors of angiotensin-converting enzyme. Carboxyalkanoyl and mercaptoalkanoyl amino acids (1977) Biochemistry, 16, pp. 5484-5491Diniz, Separação de proteínas e caracterização de substâncias ativas em venenos de aranhas do Brasil (1963) An. Acad. Bras. Sci., 35, pp. 283-291Entwistle, Johnstone, Medzihradszky, May, Isolation of a pure toxic polypeptide from the venom of the spider Phoneutria nigriventer and its neurophysiological activity on an insect femur preparation (1982) Toxicon, 20, pp. 1059-1067Fontana, Vital Brazil, Mode of action of Phoneutria nigriventer spider venom at the isolated phrenic nerve-diaphragm of the rat (1985) Braz. J. Med. Biol. Res., 18, pp. 557-565Fuller, Clements, Whitfield, Funder, Kallikrein gene expression in the rat anterior pituitary (1985) Mol. Cell. Endocrin., 39, pp. 99-105Jong, Norment, Heitz, Separation and characterization of venom components in Loxosceles reclusa—II. Protease enzyme activity (1979) Toxicon, 17, pp. 529-537Lazure, Leduc, Seidah, Chretien, Dube, Chapdelaine, Frenette, Tremblay, The major androgen-dependent protease in dog prostate belongs to the kallikrein family: confirmation by partial amino acid sequencing (1984) FEBS Lett., 175, pp. 1-7Lieberthal, Oza, Bernard, Levinsky, The effect of cations on the activity of human urinary kallikrein (1982) J. biol. Chem., 257, pp. 10,827-10,830Lucas, Spiders in Brazil (1988) Toxicon, 26, pp. 759-772Marangoni, Antunes, Brain, de Nucci, Phoneutria nigriventer (armed spider) venom activates tissue kallikrein-kininogen-kinin system in rabbit skin in vivo (1993) Br. J. Pharmac., 109, pp. 539-543Marangoni, Borges, Marangoni, Antunes, Vieira, Novello, Domont, de Nucci, Biochemical characterization of a vascular smooth muscle contracting polypeptide purified from Phoneutria nigriventer (armed spider) venom (1993) Toxicon, 31, pp. 377-384Margolius, Tissue kallikreins and kinins: regulation and roles in hypertensive and diabetic diseases (1989) A. Rev. Pharmac. Toxicol., 29, pp. 343-364Perret, Proteolytic activity of Tarantula venoms due to contamination with saliva (1977) Toxicon, 15, pp. 505-510Powers, Nasjletti, A novel kinin-generating protease (kininogenase) in the porcine anterior pituitary (1982) J. biol. Chem., 257, pp. 5594-5600Powers, Nasjletti, A kininogenase resembling glandular kallikrein in the rat pituitary pars intermedia (1983) Endocrinology, 112, pp. 1194-1200Proud, Kaplan, Kinin formation: mechanisms and role in inflammatory disorders (1988) A. Rev. Immunol., 6, pp. 49-83Proud, Bailey, Nustad, Gautvik, The immunological similarity of rat glandular kallikreins (1977) Biochem. J., 167, pp. 835-838Rezende, Jr, Cordeiro, Oliveira, Diniz, Isolation of neurotoxic peptides from the venom of the ‘armed spider’ Phoneutria nigriventer (1991) Toxicon, 29, pp. 1225-1233Schachter, Peret, Billing, Wheeler, Longridge, Immunolocalization of the protease kallikrein in the colon (1983) J. Histochem. Cytochem., 31, pp. 1255-1260Schenberg, Pereira-Lima, Pharmacology of the polypeptides from the venom of the spider Phoneutria fera (1966) Mem. Inst. Butantan, 33, pp. 627-638Schenberg, Pereira-Lima, Phoneutria nigriventer venom (1971) Pharmacology and biochemistry of its components, 3, pp. 279-297. , W. Bucherl, E.E. Buckley, Venomous Animals and Their Venoms, Academic Press, New YorkSpector, Willoughby, Endogenous mediators of increased vascular permeability in inflammation (1968) The Pharmacology of Inflammation, pp. 22-54. , English Universities PressVavrek, Stewart, Competitive antagonists of bradykinin (1985) Peptides, 6, pp. 161-164Vital Brazil, Leite, Fontana, Modo de ação da peçonha da aranha armadeira, Phoneutria nigriventer (Keyserling, 1891), nas aurículas isoladas de cobaia (1988) Cien̂c. Cul. S. Paulo, 40, pp. 181-185Vogel, Kallikrein inhibitors (1979) Bradykinin, Kallidin and Kallikrein. Handbook of Experimental Pharmacology, 25, pp. 163-225. , Springer, BerlinWilliams, Prostaglandin E2, prostaglandin I2 and the vascular changes of inflammation (1979) Br. J. Pharmac., 65, pp. 517-524Williams, Morley, Prostaglandins as potentiators of increased vascular permeability in inflammation (1973) Nature, 246, pp. 215-21

The effect of Phoneutria nigriventer (armed spider) venom on arterial blood pressure of anaesthetised rats

CNPQ – CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOThe changes induced in the mean arterial blood pressure of anaesthetised rats following the administration of armed spider (Phoneutria nigrinventer) venom have been investigated. The intravenous injection of Phoneuhria nigriventer venom (0.1 mg/kg) evoked a brief and reversible decrease in the mean arterial blood pressure whereas a higher dose of venom (0.3 mg/kg) caused a biphasic response characterized by a short-lasting hypotension followed by a sustained and prolonged hypertension (40-50 min). These changes were accompanied by tachycardia, salivation, fasciculations, defecation and respiratory disturbances. Pretreatment of the animals with atropine (10 mg/kg), propranolol (100 mg/kg), phenoxybenzamine (100 mg/kg) and indomethacin (4 mg/kg) did not significantly affect the mean arterial blood pressure changes induced by Phoneutria nigriventer venom. Similarly, the bradykinin B-2 receptor antagonist Hoe 140 (D-Arg-[Hyp(3),Thi(5),DTic(7),Oic(8)]-bradykinin (0.6 mg/kg), the PAF receptor antagonist WEB 2086 (3-(4-(2-chlorophenyl)-9-methyl-6 H-thieno-(3,2f) (1,2,4)-triazolo-(4,3-a) (1,4)aiazepine-2-yl)-(4-morpholinyl)-1-propanone) (20 mg/kg), the tachykinin NK2 receptor antagonist SR 140333 ((S)1-(2-[3-(3,4-dichlorophenyl)-1-(3-iso-propoxyphenyl acetyl) piperidin-3-yl] ethyl)-4-phenyl-1-azoniabicyclo[2.2.2] octane, chloride) (0.5 mg/kg), the tachykinin NK2 receptor antagonist SR 48968 ((S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-d (0.5 mg/kg) and the nitric oxide synthase inhibitor N-omega-nitro-L-arginine methyl ester (10 mg/kg) had no significant effect on the mean arterial blood pressure changes induced by Phoneutria nigriventer venom. The increase in the blood pressure induced by Phoneutria nigriventer venom was also not significantly affected by either the angiotensin II receptor antagonist losartan (10 mg/kg) or the endothelin ET(A) receptor antagonist FR 139317 ((R)2-[(R)-2-[[1(hexahydro-1H-azepinyl]carbonyl]amino-4-methyl-pentanoyl[amino-3-[3-(1-methyl-1H-indoyl)lpropionyl] amino-3-(2-pyridyl) propionic acid) (30 mg/kg). The ATP-dependent K+ channel antagonist glibenclamide (50 mg/kg) reduced by 40% the hypotension induced by Phoneutria nigriventer venom without affecting the hypertensive response. Pretreatment of the animals with L-type Ca2+ channel antagonists such as verapamil (10-100 mu g/kg/min), diltiazem (40-120 mu g/kg/min) and nifedipine (0.3-10 mg/kg) markedly attenuated the hypertension induced by Phoneutria nigriventer venom. Verapamil (30 mu g/kg/min) and diltiazem (120 mu g/kg/min) also promptly reversed the established hypertension induced by Phoneutria nigriventer venom when infused 8 min after venom injection. Our results indicate that the brief decrease of blood pressure induced by Phoneutria nigriventer venom.is partially due to ATP-dependent K+ channel activation. The prolonged hypertension seems to result from direct Ca2+ entry into vascular and/or cardiac muscles2982113120CNPQ – CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCNPQ – CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOsem informaçã

Comparative effect of phoneutria nigriventer spider venom and capsaicin on the rat paw oedema

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCapsaicin, the pungent component of hot peppers, and the venom of the spider Phoneutria nigriventer are able to activate sensory nerves resulting in cutaneous neurogenic plasma extravasation. This study was undertaken to compare the ability of these substances to evoke oedema in the rat hind-paw and mechanisms underlying this effect. Subplantar injection of either Phoneutria nigriventer venom (PNV; 1-100 mug/paw) or capsaicin (10-200 mug/paw) caused a significant paw oedema that was potentiated by CGRP (10 pmol/paw). In rats treated neonatally with capsaicin to deplete neuropeptides. the paw oedema induced by either PNV (100 mug/paw) or capsaicin (100 mug/paw) was partially reduced (P <0.05). The tachykinin NKI receptor antagonist SR140333 (0.2 mu mol/kg; i.v.) prevented the paw oedema induced by the tachykinin NK1 receptor agonist GR73632 (30 pmol/paw) and partially reduced paw oedema induced by PNV or capsaicin. Treatment of rats with compound 48/80 (5 mg/kg; s.c. 3 days) or with both HI receptor antagonist (mepyramine; 1 nmol/paw) and 5-HT receptor antagonist (methysergide; I nmol/paw) significantly inhibited PNV- or capsaicin-induced paw oedema. The combined treatment with mepyramine and methysergide and SR140333 further reduced PNV- and capsaicin-induced paw oedema. The bradykinin B-2 receptor antagonist Hoe 140 affected neither PNV- nor capsaicin-induced responses. Our results suggest that PNV and capsaicin each induce paw oedema that is partially mediated by activation of sensory fibers culminating in the release of substance P as well as by activation of mast cells which in turn release amines such as histamine and 5-HT691315731585FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOsem informaçã