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Made available in DSpace on 2018-11-28T21:36:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 UNESP UNIFESP Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Rhamnetin (Rhm), 3-O-methylquercetin (3MQ), and Rhamnazin (Rhz) are methylated derivatives of quercetin commonly found in fruits and vegetables that possess antioxidant and anti-inflammatory properties. Phospholipase A2 (PLA2) displays several important roles during acute inflammation; therefore, this study aimed at investigating new compounds able to inhibit this enzyme, besides evaluating creatine kinase (CK) levels and citotoxicity. Methylated quercetins were compared with quercetin (Q) and were incubated with secretory PLA2 (sPLA2) from Bothrops jararacussu to determine their inhibitory activity. Cytotoxic studies were performed by using the J774 cell lineage incubated with quercertins. In vivo tests were performed with Swiss female mice to evaluate decreasing paw edema potential and compounds' CK levels. Structural modifications on sPLA2 were made with circular dichroism (CD). Despite Q and Rhz showing greater enzymatic inhibitory potential, high CK was observed. Rhm exhibited sPLA2 inhibitory potential, no toxicity and, remarkably, it decreased CK levels. The presence of 3OH on the C-ring of Rhm may contribute to both its anti-inflammatory and enzymatic inhibition of sPLA2, and the methylation of ring A may provide the increase in cell viability and low CK level induced by sPLA2. These results showed that Rhm can be a candidate as a natural compound for the development of new anti-inflammatory drugs. Univ Fed Sao Paulo, Postgrad Program Food Nutr & Hlth, BR-11015020 Sao Paulo, Brazil Univ Estadual Paulista, Biosci Inst, BR-11330900 Sao Paulo, Brazil Brazil Univ, Prorector Res, BR-08230030 Sao Paulo, Brazil Univ Sao Paulo, Pathol Lab Infect Dis LIM50, Dept Pathol, Sch Med, BR-01246903 Sao Paulo, Brazil Univ Estadual Paulista, Biosci Inst, BR-11330900 Sao Paulo, Brazil

Made available in DSpace on 2014-12-03T13:11:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-03-01Bitstream added on 2014-12-03T13:23:33Z : No. of bitstreams: 1 WOS000316607800023.pdf: 364360 bytes, checksum: fdfdf38afecf4c0522eef5897bd5ac91 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The sulfated polysaccharides from Solieria filiformis (Sf), Botryocladia occidentalis (Bo), Caulerpa racemosa (Cr) and Gracilaria caudata (Gc) were extracted and extensively purified. These compounds were then subjected to in vitro assays to evaluate the inhibition of these polysaccharides on the growth of Leishmania (L.) amazonensis promastigotes. Under the same assay conditions, only three of the four sulfated polysaccharides were active against L. amazonensis, and the polysaccharide purified from Cr was the most potent (EC50 value: 34.5 mu g/mL). The polysaccharides derived from Bo and Sf demonstrated moderate anti-leishmanial activity (EC50 values of 63.7 mu g/mL and 137.4 mu g/mL). In addition, we also performed in vitro cytotoxic assays toward peritoneal macrophages and J774 macrophages. For the in vitro cytotoxicity assay employing J774 cells, all of the sulfated polysaccharides decreased cell survival, with CC50 values of 27.3 mu g/mL, 49.3 mu g/mL, 73.2 mu g/mL, and 99.8 mu g/mL for Bo, Cr, Gc, and Sf, respectively. However, none of the sulfated polysaccharides reduced the cell growth rate of the peritoneal macrophages. These results suggest that macroalgae contain compounds with various chemical properties that can control specific pathogens. According to our results, the assayed sulfated polysaccharides were able to modulate the growth rate and cell survival of Leishmania (L.) amazonensis promastigotes in in vitro assays, and these effects involved the interaction of the sulfated polysaccharides on the cell membrane of the parasites. State Univ Sao Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil Univ Prebiteriana Mackenzie, Ctr Biol & Hlth Sci, BR-01302907 Sao Paulo, Brazil Univ Fed Ceara, Pici Unit, UFC, BR-60356000 Fortaleza, Ceara, Brazil State Univ Sao Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil

Made available in DSpace on 2013-08-28T14:13:53Z (GMT). No. of bitstreams: 1 WOS000308300500001.pdf: 5416562 bytes, checksum: 067c932d70822bc39bb0b2a57e0ab891 (MD5) Made available in DSpace on 2013-09-30T17:41:27Z (GMT). No. of bitstreams: 2 WOS000308300500001.pdf: 5416562 bytes, checksum: 067c932d70822bc39bb0b2a57e0ab891 (MD5) WOS000308300500001.pdf.txt: 36390 bytes, checksum: 647f2b3b8e3fb1f7f5eb07947fc6a27f (MD5) Previous issue date: 2012-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:12:24Z No. of bitstreams: 2 WOS000308300500001.pdf: 5416562 bytes, checksum: 067c932d70822bc39bb0b2a57e0ab891 (MD5) WOS000308300500001.pdf.txt: 36390 bytes, checksum: 647f2b3b8e3fb1f7f5eb07947fc6a27f (MD5) Made available in DSpace on 2014-05-20T13:12:24Z (GMT). No. of bitstreams: 2 WOS000308300500001.pdf: 5416562 bytes, checksum: 067c932d70822bc39bb0b2a57e0ab891 (MD5) WOS000308300500001.pdf.txt: 36390 bytes, checksum: 647f2b3b8e3fb1f7f5eb07947fc6a27f (MD5) Previous issue date: 2012-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) Secretory phospholipases A(2) (sPLA(2)) exert proinflammatory actions through lipid mediators. These enzymes have been found to be elevated in many inflammatory disorders such as rheumatoid arthritis, sepsis, and atherosclerosis. The aim of this study was to evaluate the effect of harpalycin 2 (Har2), an isoflavone isolated from Harpalyce brasiliana Benth., in the enzymatic, edematogenic, and myotoxic activities of sPLA2 from Bothrops pirajai, Crotalus durissus terrificus, Apis mellifera, and Naja naja venoms. Har2 inhibits all sPLA(2) tested. PrTX-III (B. pirajai venom) was inhibited at about 58.7%, Cdt F15 (C. d. terrificus venom) at 78.8%, Apis (from bee venom) at 87.7%, and Naja (N. naja venom) at 88.1%. Edema induced by exogenous sPLA(2) administration performed in mice paws showed significant inhibition by Har2 at the initial step. In addition, Har2 also inhibited the myotoxic activity of these sPLA(2)s. In order to understand how Har2 interacts with these enzymes, docking calculations were made, indicating that the residues His48 and Asp49 in the active site of these enzymes interacted powerfully with Har2 through hydrogen bonds. These data pointed to a possible anti-inflammatory activity of Har2 through sPLA(2) inhibition. Univ Fed Ceara, Dept Fisiol & Farmacol, BR-60430270 Fortaleza, CE, Brazil Universidade Federal de Pernambuco (UFPE), Dept Ciencias Farmaceut, BR-50740520 Recife, PE, Brazil Univ Fed Rio Grande do Norte, Dept Quim, BR-50078970 Natal, RN, Brazil Univ Fed Ceara, Ctr NE Aplicacao & Uso Ressonancia Magnet Nucl CE, BR-60455760 Fortaleza, CE, Brazil Univ Estadual Campinas, Inst Biomed, Dept Bioquim, BR-13082862 Campinas, SP, Brazil Univ Estadual Paulista, Unidade Sao Vicente, BR-11330900 Sao Vicente, SP, Brazil Univ Presbiteriana Mackenzie, Ctr Ciencias Biol & Saude, BR-01302970 São Paulo, Brazil Univ Estadual Paulista, Unidade Sao Vicente, BR-11330900 Sao Vicente, SP, Brazil

Made available in DSpace on 2015-03-18T15:55:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01Bitstream added on 2015-03-18T16:28:37Z : No. of bitstreams: 1 WOS000345403500001.epub: 3212256 bytes, checksum: ee5ec134914559e8861623b02dbfface (MD5)Bitstream added on 2015-03-18T16:28:37Z : No. of bitstreams: 2 WOS000345403500001.epub: 3212256 bytes, checksum: ee5ec134914559e8861623b02dbfface (MD5) WOS000345403500001.pdf: 3025372 bytes, checksum: f114a4680e7dd3732e5006545984e266 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundo Mackenzie de Pesquisa Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The aim of this paper was to investigate the effect of chlorogenic acid (5-caffeoylquinic acid, 5CQA), isolated from Baccharis oxyodonta, on the structure and pharmacological effect of secretory phospholipase A2 (sPLA2) from Crotalus durissus terrificus. All in vitro and in vivo experiments were conducted using a purified sPLA2 compared under the same experimental conditions with sPLA2 : 5CQA. 5CQA induced several discrete modifications in the secondary structure and the hydrophobic characteristics of native sPLA2 that induced slight changes in the alpha-helical content, increase in the randomcoil structure, and decrease of fluorescence of native sPLA2. Moreover, 5CQA significantly decreased the enzymatic activity and the oedema and myonecrosis induced by native sPLA2. As the catalytic activity of sPLA2 plays an important role in several of its biological and pharmacological properties, antibacterial activity was used to confirm the decrease in its enzymatic activity by 5CQA, which induced massive bacterial cell destruction. We found that 5CQA specifically abolished the enzymatic activity of sPLA2 and induced discrete protein unfolding that mainly involved the pharmacological site of sPLA2. These results showed the potential application of 5CQA in the snake poisoning treatment and modulation of the pathological effect of inflammation induced by secretory PLA2. UNESP, BR-11330900 Sao Vicente, SP, Brazil Univ Sao Paulo, Inst Biociencias, Dept Bot, BR-05508090 Sao Paulo, Brazil Univ Presbiteriana Mackenzie, Escola Engn, Curso Quim, BR-01302907 Sao Paulo, Brazil Univ Presbiteriana Mackenzie, Ctr Ciencias Biol & Saude, Curso Ciencias Biol, BR-01302907 Sao Paulo, Brazil UNESP, BR-11330900 Sao Vicente, SP, Brazil FAPESP: 11/06704-4

Made available in DSpace on 2016-04-01T18:45:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2014. Added 1 bitstream(s) on 2016-04-01T18:50:17Z : No. of bitstreams: 1 ISSN1110-7243-2014-2014-01-11.pdf: 1666043 bytes, checksum: ac41ab698ee781e99d5f25a508548554 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Instituto Nacional para Pesquisa em Toxinas (INCT-Tox) This paper shows the results of quercitrin effects on the structure and biological activity of secretory phospholipase (sPLA2) from Crotalus durissus terrificus, which is the main toxin involved in the pharmacological effects of this snake venom. According to our mass spectrometry and circular dichroism results, quercetin was able to promote a chemical modification of some amino acid residues and modify the secondary structure of C. d. terrificus sPLA2. Moreover, molecular docking studies showed that quercitrin can establish chemical interactions with some of the crucial amino acid residues involved in the enzymatic activity of the sPLA2, indicating that this flavonoid could also physically impair substrate molecule access to the catalytic site of the toxin. Additionally, in vitro and in vivo assays showed that the quercitrin strongly diminished the catalytic activity of the protein, altered its Vmax and Km values, and presented a more potent inhibition of essential pharmacological activities in the C. d. terrificus sPLA2, such as its myotoxicity and edematogenic effect, in comparison to quercetin. Thus, we concluded that the rhamnose group found in quercitrin is most likely essential to the antivenom activities of this flavonoid against C. d. terrificus sPLA2. Universidade Presbiteriana Mackenzie, Centro de Ciências Biológicas e da Saúde (CCBS), São Paulo, SP, Brasil Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Instituto de Biociências, Departamento de Ciências Biológicas, São Vicente, SP, Brasil Universidade Estadual de Campinas (UNICAMP), Faculdade de Ciências Médicas, Campinas, SP, Brasil Universidade Presbiteriana Mackenzie, Escola de Engenharia, São Paulo, SP, Brasil Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Instituto de Biociências de Botucatu (IBB), Departamento de Física e Biofísica, Botucatu, SP, Brasil Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Instituto de Biociências, Departamento de Ciências Biológicas, São Vicente, SP, Brasil Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Instituto de Biociências de Botucatu (IBB), Departamento de Física e Biofísica, Botucatu, SP, Brasil FAPESP: 2011/06704-4 FAPESP: 2012/06502-5 FAPESP: 2013/12077-8

Made available in DSpace on 2014-10-01T13:08:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-01Bitstream added on 2014-10-01T14:03:46Z : No. of bitstreams: 1 S0102-695X2013000400010.pdf: 833708 bytes, checksum: f93863ef378ba4817367d374e463cefd (MD5) Sulfated polysaccharides derived from seaweed have shown great potential for use in the development of new drugs. In this study, we observed that a low-molecular-weight sulfated polysaccharide from Caulerpa racemosa, termed CrSP, could interact with secretory phospholipase A2 (sPLA2) isolated from Crotalus durissus terrificus venom. When native sPLA2 (14 kDa) was incubated with CrSP, they formed a molecular complex (sPLA2:CrSP) with a molecular mass of 32 kDa, approximately. Size exclusion chromatography experiments suggested that CrSP formed a stable complex with sPLA2. We belived that sPLA2 and SPCr are involved an ionic interaction between negatively charged CrSP and the positively charged basic amino acid residues of sPLA2, because this interaction induced significant changes in sPLA2 enzymatic and pharmacological activities. CrSP caused a significant increase in sPLA2 enzymatic and bactericidal activity and increased its edematogenic effect. A pharmacological assay showed that the myotoxic activity of sPLA2:CrSP is unrelated to its enzymatic activity and that sPLA2:CrSP may have a practical application as a natural antibacterial agent for use in humans and commercially raised animals. Universidade Estadual Paulista Laboratório de Biologia Molecular e Peptídeos Universidade Presbiteriana Mackenzie Centro de Ciências Biológicas e da Saúde Universidade Federal do Ceará Centro de Ciências Agrárias Departamento de Engenharia de Pesca Universidade Estadual Paulista Laboratório de Biologia Molecular e Peptídeos

Made available in DSpace on 2013-08-28T14:14:23Z (GMT). No. of bitstreams: 1 WOS000312325000001.pdf: 1687045 bytes, checksum: 81c838bb86f5cc0e9eecf68046ea6082 (MD5) Made available in DSpace on 2013-09-30T17:41:27Z (GMT). No. of bitstreams: 2 WOS000312325000001.pdf: 1687045 bytes, checksum: 81c838bb86f5cc0e9eecf68046ea6082 (MD5) WOS000312325000001.pdf.txt: 42001 bytes, checksum: b4a008ad203891063588dc25c979630f (MD5) Previous issue date: 2012-08-27 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:12:24Z No. of bitstreams: 2 WOS000312325000001.pdf: 1687045 bytes, checksum: 81c838bb86f5cc0e9eecf68046ea6082 (MD5) WOS000312325000001.pdf.txt: 42001 bytes, checksum: b4a008ad203891063588dc25c979630f (MD5) Made available in DSpace on 2014-05-20T13:12:24Z (GMT). No. of bitstreams: 2 WOS000312325000001.pdf: 1687045 bytes, checksum: 81c838bb86f5cc0e9eecf68046ea6082 (MD5) WOS000312325000001.pdf.txt: 42001 bytes, checksum: b4a008ad203891063588dc25c979630f (MD5) Previous issue date: 2012-08-27 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) Background: Harpalycin 2 (HP-2) is an isoflavone isolated from the leaves of Harpalyce brasiliana Benth., a snakeroot found in northeast region of Brazil and used in folk medicine to treat snakebite. Its leaves are said to be anti-inflammatory. Secretory phospholipases A(2) are important toxins found in snake venom and are structurally related to those found in inflammatory conditions in mammals, as in arthritis and atherosclerosis, and for this reason can be valuable tools for searching new anti-phospholipase A(2) drugs.Methods: HP-2 and piratoxin-III (PrTX-III) were purified through chromatographic techniques. The effect of HP-2 in the enzymatic activity of PrTX-III was carried out using 4-nitro-3-octanoyloxy-benzoic acid as the substrate. PrTX-III induced platelet aggregation was inhibited by HP-2 when compared to aristolochic acid and p-bromophenacyl bromide (p-BPB). In an attempt to elucidate how HP-2 interacts with PrTX-III, mass spectrometry, circular dichroism and intrinsic fluorescence analysis were performed. Docking scores of the ligands (HP-2, aristolochic acid and p-BPB) using PrTX-III as target were also calculated.Results: HP-2 inhibited the enzymatic activity of PrTX-III (IC50 11.34 +/- 0.28 mu g/mL) although it did not form a stable chemical complex in the active site, since mass spectrometry measurements showed no difference between native (13,837.34 Da) and HP-2 treated PrTX-III (13,856.12 Da). A structural analysis of PrTX-III after treatment with HP-2 showed a decrease in dimerization and a slight protein unfolding. In the platelet aggregation assay, HP-2 previously incubated with PrTX-III inhibited the aggregation when compared with untreated protein. PrTX-III chemical treated with aristolochic acid and p-BPB, two standard PLA(2) inhibitors, showed low inhibitory effects when compared with the HP-2 treatment. Docking scores corroborated these results, showing higher affinity of HP-2 for the PrTX-III target (PDB code: 1GMZ) than aristolochic acid and p-BPB. HP-2 previous incubated with the platelets inhibits the aggregation induced by untreated PrTX-III as well as arachidonic acid.Conclusion: HP-2 changes the structure of PrTX-III, inhibiting the enzymatic activity of this enzyme. In addition, PrTX-III platelet aggregant activity was inhibited by treatment with HP-2, p-BPB and aristolochic acid, and these results were corroborated by docking scores. State Univ São Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil Univ Fed Ceara, UFC, Dept Physiol & Pharmacol, BR-60430270 Fortaleza, Ceara, Brazil Univ Fed Ceara, UFC, Dept Clin & Toxicol Anal, BR-60430370 Fortaleza, Ceara, Brazil Univ Fed Rio Grande do Norte, UFRN, Dept Chem, BR-50078970 Natal, RN, Brazil Univ Fed Ceara, Dept Organ & Inorgan Chem, UFC, BR-60455760 Fortaleza, Ceara, Brazil Universidade Federal de Pernambuco (UFPE), UFPE, Dept Pharmaceut Sci, BR-50740520 Recife, PE, Brazil Univ Estadual Campinas, UNICAMP, Inst Biol, Dept Biochem, BR-13082862 Campinas, SP, Brazil Univ Prebiteriana Mackenzie, Ctr Biol & Hlth Sci, BR-01302970 São Paulo, Brazil State Univ São Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil