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Douglas Oscar Ceolin Mariano
Laboratório de Bioquímica e Biofísica, Instituto Butantan, 05503-900 São Paulo, Brazil

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Journal article
Published: 11 May 2021 in Molecules
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Kynurenic acid (KYNA) is derived from tryptophan, formed by the kynurenic pathway. KYNA is being widely studied as a biomarker for neurological and cardiovascular diseases, as it is found in ischemic conditions as a protective agent; however, little is known about its effect after ischemia-reperfusion in the vascular system. We induced ischemia for 30 min followed by 5 min reperfusion (I/R) in the rat aorta for KYNA evaluation using functional assays combined with proteomics. KYNA recovered the exacerbated contraction induced by phenylephrine and relaxation induced by acetylcholine or sodium nitroprussiate in the I/R aorta, with vessel responses returning to values observed without I/R. The functional recovery can be related to the antioxidant activity of KYNA, which may be acting on the endothelium-injury prevention, especially during reperfusion, and to proteins that regulate neurotransmission and cell repair/growth, expressed after the KYNA treatment. These proteins interacted in a network, confirming a protein profile expression for endothelium and neuron repair after I/R. Thus, the KYNA treatment had the ability to recover the functionality of injured ischemic-reperfusion aorta, by tissue repairing and control of neurotransmitter release, which reinforces its role in the post-ischemic condition, and can be useful in the treatment of such disease.

ACS Style

Viviane Lima; Douglas Mariano; Hugo Vigerelli; Sabrina Janussi; Thayz Baptista; Mário Claudino; Daniel Pimenta; Juliana Sciani. Effects of Kynurenic Acid on the Rat Aorta Ischemia—Reperfusion Model: Pharmacological Characterization and Proteomic Profiling. Molecules 2021, 26, 2845 .

AMA Style

Viviane Lima, Douglas Mariano, Hugo Vigerelli, Sabrina Janussi, Thayz Baptista, Mário Claudino, Daniel Pimenta, Juliana Sciani. Effects of Kynurenic Acid on the Rat Aorta Ischemia—Reperfusion Model: Pharmacological Characterization and Proteomic Profiling. Molecules. 2021; 26 (10):2845.

Chicago/Turabian Style

Viviane Lima; Douglas Mariano; Hugo Vigerelli; Sabrina Janussi; Thayz Baptista; Mário Claudino; Daniel Pimenta; Juliana Sciani. 2021. "Effects of Kynurenic Acid on the Rat Aorta Ischemia—Reperfusion Model: Pharmacological Characterization and Proteomic Profiling." Molecules 26, no. 10: 2845.

Research
Published: 01 January 2021 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed.

ACS Style

Giselle Pacifico Sartori; Andréa da Costa; Fernanda Lúcio dos Santos Macarini; Douglas Oscar Ceolin Mariano; Daniel Carvalho Pimenta; Patrick Jack Spencer; Luiz Henrique Da Silva Nali; Andrés Jimenez Galisteo Jr.. Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation. Journal of Venomous Animals and Toxins including Tropical Diseases 2021, 27, 1 .

AMA Style

Giselle Pacifico Sartori, Andréa da Costa, Fernanda Lúcio dos Santos Macarini, Douglas Oscar Ceolin Mariano, Daniel Carvalho Pimenta, Patrick Jack Spencer, Luiz Henrique Da Silva Nali, Andrés Jimenez Galisteo Jr.. Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation. Journal of Venomous Animals and Toxins including Tropical Diseases. 2021; 27 ():1.

Chicago/Turabian Style

Giselle Pacifico Sartori; Andréa da Costa; Fernanda Lúcio dos Santos Macarini; Douglas Oscar Ceolin Mariano; Daniel Carvalho Pimenta; Patrick Jack Spencer; Luiz Henrique Da Silva Nali; Andrés Jimenez Galisteo Jr.. 2021. "Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation." Journal of Venomous Animals and Toxins including Tropical Diseases 27, no. : 1.

Journal article
Published: 29 July 2019 in Toxins
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Ant species have specialized venom systems developed to sting and inoculate a biological cocktail of organic compounds, including peptide and polypeptide toxins, for the purpose of predation and defense. The genus Dinoponera comprises predatory giant ants that inoculate venom capable of causing long-lasting local pain, involuntary shaking, lymphadenopathy, and cardiac arrhythmias, among other symptoms. To deepen our knowledge about venom composition with regard to protein toxins and their roles in the chemical–ecological relationship and human health, we performed a bottom-up proteomics analysis of the crude venom of the giant ant D. quadriceps, popularly known as the “false” tocandiras. For this purpose, we used two different analytical approaches: (i) gel-based proteomics approach, wherein the crude venom was resolved by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and all protein bands were excised for analysis; (ii) solution-based proteomics approach, wherein the crude venom protein components were directly fragmented into tryptic peptides in solution for analysis. The proteomic data that resulted from these two methodologies were compared against a previously annotated transcriptomic database of D. quadriceps, and subsequently, a homology search was performed for all identified transcript products. The gel-based proteomics approach unequivocally identified nine toxins of high molecular mass in the venom, as for example, enzymes [hyaluronidase, phospholipase A1, dipeptidyl peptidase and glucose dehydrogenase/flavin adenine dinucleotide (FAD) quinone] and diverse venom allergens (homologous of the red fire ant Selenopsis invicta) and venom-related proteins (major royal jelly-like). Moreover, the solution-based proteomics revealed and confirmed the presence of several hydrolases, oxidoreductases, proteases, Kunitz-like polypeptides, and the less abundant inhibitor cysteine knot (ICK)-like (knottin) neurotoxins and insect defensin. Our results showed that the major components of the D. quadriceps venom are toxins that are highly likely to damage cell membranes and tissue, to cause neurotoxicity, and to induce allergic reactions, thus, expanding the knowledge about D. quadriceps venom composition and its potential biological effects on prey and victims.

ACS Style

Douglas Oscar Ceolin Mariano; Úrsula Castro De Oliveira; André Junqueira Zaharenko; Daniel Carvalho Pimenta; Gandhi Rádis-Baptista; Álvaro Rossan De Brandão Prieto-Da-Silva. Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps. Toxins 2019, 11, 448 .

AMA Style

Douglas Oscar Ceolin Mariano, Úrsula Castro De Oliveira, André Junqueira Zaharenko, Daniel Carvalho Pimenta, Gandhi Rádis-Baptista, Álvaro Rossan De Brandão Prieto-Da-Silva. Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps. Toxins. 2019; 11 (8):448.

Chicago/Turabian Style

Douglas Oscar Ceolin Mariano; Úrsula Castro De Oliveira; André Junqueira Zaharenko; Daniel Carvalho Pimenta; Gandhi Rádis-Baptista; Álvaro Rossan De Brandão Prieto-Da-Silva. 2019. "Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps." Toxins 11, no. 8: 448.

Journal article
Published: 31 March 2019 in Toxins
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Although omics studies have indicated presence of proteases on the Tityus serrulatus venom (TsV), little is known about the function of these molecules. The TsV contains metalloproteases that cleave a series of human neuropeptides, including the dynorphin A (1-13) and the members of neuropeptide Y family. Aiming to isolate the proteases responsible for this activity, the metalloserrulase 3 and 4 (TsMS 3 and TsMS 4) were purified after two chromatographic steps and identified by mass spectrometry analysis. The biochemical parameters (pH, temperature and cation effects) were determined for both proteases, and the catalytic parameters (Km, k cat , cleavage sites) of TsMS 4 over fluorescent substrate were obtained. The metalloserrulases have a high preference for cleaving neuropeptides but presented different primary specificities. For example, the Leu-enkephalin released from dynorphin A (1-13) hydrolysis was exclusively performed by TsMS 3. Neutralization assays using Butantan Institute antivenoms show that both metalloserrulases were well blocked. Although TsMS 3 and TsMS 4 were previously described through cDNA library studies using the venom gland, this is the first time that both these toxins were purified. Thus, this study represents a step further in understanding the mechanism of scorpion venom metalloproteases, which may act as possible neuropeptidases in the envenomation process.

ACS Style

Daniela Cajado-Carvalho; Cristiane Castilho Fernandes Da Silva; Roberto Tadashi Kodama; Douglas Oscar Ceolin Mariano; Daniel Carvalho Pimenta; Bruno Duzzi; Alexandre Kazuo Kuniyoshi; Fernanda Vieira Portaro. Purification and Biochemical Characterization of TsMS 3 and TsMS 4: Neuropeptide-Degrading Metallopeptidases in the Tityus serrulatus Venom. Toxins 2019, 11, 194 .

AMA Style

Daniela Cajado-Carvalho, Cristiane Castilho Fernandes Da Silva, Roberto Tadashi Kodama, Douglas Oscar Ceolin Mariano, Daniel Carvalho Pimenta, Bruno Duzzi, Alexandre Kazuo Kuniyoshi, Fernanda Vieira Portaro. Purification and Biochemical Characterization of TsMS 3 and TsMS 4: Neuropeptide-Degrading Metallopeptidases in the Tityus serrulatus Venom. Toxins. 2019; 11 (4):194.

Chicago/Turabian Style

Daniela Cajado-Carvalho; Cristiane Castilho Fernandes Da Silva; Roberto Tadashi Kodama; Douglas Oscar Ceolin Mariano; Daniel Carvalho Pimenta; Bruno Duzzi; Alexandre Kazuo Kuniyoshi; Fernanda Vieira Portaro. 2019. "Purification and Biochemical Characterization of TsMS 3 and TsMS 4: Neuropeptide-Degrading Metallopeptidases in the Tityus serrulatus Venom." Toxins 11, no. 4: 194.

Journal article
Published: 30 March 2019 in Archives of Biochemistry and Biophysics
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In addition to autophagy, proteasomes are critical for regulating intracellular protein levels and removing misfolded proteins. The 20S proteasome (20SPT), the central catalytic unit, is sometimes flanked by regulatory units at one or both ends. Additionally, proteosomal activation has been associated with increased lifespan in many organisms. Our group previously reported that the gating (open/closed) of the free 20S proteasome is redox controlled, and that S-glutathionylation of two Cys residues (Cys76 and Cys221) in the α5 subunit promotes gate opening. The present study constructed site-directed mutants of these Cys residues, and evaluated the effects these mutations have on proteosome gate opening and yeast cell survival. Notably, the double mutation of both Cys residues (Cys76 and Cys221) rendered the cells nonviable, whereas the lifespan of the yeast carrying the single mutations (α5-C76S or α5-C221S) was attenuated when compared to the wild type counterpart. Furthermore, it was found that α5-C76S or α5-C221S 20SPT were more likely to be found with the gate in a closed conformation. In contrast, a random α5-subunit double mutation (S35P/C221S) promoted gate opening, increased chronological lifespan and provided resistance to oxidative stress. The 20SPT core particle purified from the long-lived strain degraded model proteins (e.g., α-synuclein) more efficiently than preparations obtained from the wild-type counterpart, and also displayed an increased chymotrypsin-like activity. Mass spectrometric analyses of the C76S, C221S, S35P/C221S, S35P and S35P/C76S mutants provided evidence that the highly conserved Cys76 residue of the α5-subunit is the key determinant for gate opening and cellular survival. The present study reveals a sophisticated regulatory mechanism that controls gate opening, which appears to be based on the interactions among multiple residues within the α5-subunit, and consequently impacts the lifespan of yeast.

ACS Style

Janaína M.M. Leme; Erina Ohara; Verônica Feijoli Santiago; Mario Barros; Luis E.S. Netto; Daniel C. Pimenta; Douglas Mariano; Cristiano L.P. Oliveira; Renata N. Bicev; Maria L.M. Barreto-Chaves; Caroline A. Lino; Marilene Demasi. Mutations of Cys and Ser residues in the α5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespan. Archives of Biochemistry and Biophysics 2019, 666, 63 -72.

AMA Style

Janaína M.M. Leme, Erina Ohara, Verônica Feijoli Santiago, Mario Barros, Luis E.S. Netto, Daniel C. Pimenta, Douglas Mariano, Cristiano L.P. Oliveira, Renata N. Bicev, Maria L.M. Barreto-Chaves, Caroline A. Lino, Marilene Demasi. Mutations of Cys and Ser residues in the α5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespan. Archives of Biochemistry and Biophysics. 2019; 666 ():63-72.

Chicago/Turabian Style

Janaína M.M. Leme; Erina Ohara; Verônica Feijoli Santiago; Mario Barros; Luis E.S. Netto; Daniel C. Pimenta; Douglas Mariano; Cristiano L.P. Oliveira; Renata N. Bicev; Maria L.M. Barreto-Chaves; Caroline A. Lino; Marilene Demasi. 2019. "Mutations of Cys and Ser residues in the α5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespan." Archives of Biochemistry and Biophysics 666, no. : 63-72.

Journal article
Published: 01 January 2019 in JBRA Assisted Reproduction
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The follicular fluid (FF) of women with polycystic ovary syndrome (PCOS) seems to exhibit a profile different from that of fertile women, which may be related to folliculogenesis disruption in PCOS patients. The aim of this study was to evaluate the differentially expressed proteins in the FF of women with PCOS compared to oocyte donors (ODs). This screening study included thirteen (13) women who underwent in vitro fertilization (IVF) cycles: seven (7) ODs and six (6) PCOS patients. The patients underwent standard ovarian stimulation, and the FF was analysed using ion trap and time-of-flight liquid chromatography-mass spectrometry (LCMS-IT-TOF). The FF of the patients was matched to 229 proteins, with 61 proteins exclusive to the PCOS group, 123 proteins exclusive to the ODs, and 45 proteins found in both groups. We highlight fetuin-A and vitamin D ligand protein, which were exclusively expressed in the PCOS group; Complement C3 overexpressed in the PCOS group; and 26S protease only expressed in the OD group. The canonical pathways LXR/RXR activation, FXR/RXR activation, prothrombin activation are directly related to the disrupted metabolism and increased inflammatory status found in PCOS patients. The findings of the differentially expressed proteins and matched pathways are associated with folliculogenesis, indicating it relevance to oocyte quality.

ACS Style

Thais S Domingues; Tatiana Cs Bonetti; Daniel C Pimenta; Douglas O C Mariano; Bruna Barros; Ana Paula Aquino; Eduardo L A Motta. Proteomic profile of follicular fluid from patients with polycystic ovary syndrome (PCOS) submitted to in vitro fertilization (IVF) compared to oocyte donors. JBRA Assisted Reproduction 2019, 23, 367 -391.

AMA Style

Thais S Domingues, Tatiana Cs Bonetti, Daniel C Pimenta, Douglas O C Mariano, Bruna Barros, Ana Paula Aquino, Eduardo L A Motta. Proteomic profile of follicular fluid from patients with polycystic ovary syndrome (PCOS) submitted to in vitro fertilization (IVF) compared to oocyte donors. JBRA Assisted Reproduction. 2019; 23 (4):367-391.

Chicago/Turabian Style

Thais S Domingues; Tatiana Cs Bonetti; Daniel C Pimenta; Douglas O C Mariano; Bruna Barros; Ana Paula Aquino; Eduardo L A Motta. 2019. "Proteomic profile of follicular fluid from patients with polycystic ovary syndrome (PCOS) submitted to in vitro fertilization (IVF) compared to oocyte donors." JBRA Assisted Reproduction 23, no. 4: 367-391.

Journal article
Published: 01 January 2019 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Bufonid parotoid macrogland secretion contains several low molecular mass molecules, such as alkaloids and steroids. Nevertheless, its protein content is poorly understood. Herein, we applied a sample preparation methodology that allows the analysis of viscous matrices in order to examine its proteins. Duttaphrynus melanostictus parotoid macrogland secretion was submitted to ion-exchange batch sample preparation, yielding two fractions: salt-displaced fraction and acid-displaced fraction. Each sample was then fractionated by anionic-exchange chromatography, followed by in-solution proteomic analysis. Forty-two proteins could be identified, such as acyl-CoA-binding protein, alcohol dehydrogenase, calmodulin, galectin and histone. Moreover, de novo analyses yielded 153 peptides, whereas BLAST analyses corroborated some of the proteomic-identified proteins. Furthermore, the de novo peptide analyses indicate the presence of proteins related to apoptosis, cellular structure, catalysis and transport processes. Proper sample preparation allowed the proteomic and de novo identification of different proteins in the D. melanostictus parotoid macrogland secretion. These results may increase the knowledge about the universe of molecules that compose amphibian skin secretion, as well as to understand their biological/physiological role in the granular gland.

ACS Style

Douglas Oscar Ceolin Mariano; Marcela Di Giacomo Messias; Patrick Jack Spencer; Daniel Carvalho Pimenta. Protein identification from the parotoid macrogland secretion of Duttaphrynus melanostictus. Journal of Venomous Animals and Toxins including Tropical Diseases 2019, 25, e20190029 .

AMA Style

Douglas Oscar Ceolin Mariano, Marcela Di Giacomo Messias, Patrick Jack Spencer, Daniel Carvalho Pimenta. Protein identification from the parotoid macrogland secretion of Duttaphrynus melanostictus. Journal of Venomous Animals and Toxins including Tropical Diseases. 2019; 25 ():e20190029.

Chicago/Turabian Style

Douglas Oscar Ceolin Mariano; Marcela Di Giacomo Messias; Patrick Jack Spencer; Daniel Carvalho Pimenta. 2019. "Protein identification from the parotoid macrogland secretion of Duttaphrynus melanostictus." Journal of Venomous Animals and Toxins including Tropical Diseases 25, no. : e20190029.

Journal article
Published: 19 June 2018 in Toxins
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Scorpion venoms are composed of several substances with different pharmacological activities. Neurotoxins exert their effects by targeting ion channels resulting in toxic effects to mammals, insects and crustaceans. Tb II-I, a fraction isolated from Tityus bahiensis scorpion venom, was investigated for its ability to induce neurological and immune-inflammatory effects. Two putative β-sodium channel toxins were identified in this fraction, Tb2 II and Tb 4, the latter having been completely sequenced by mass spectrometry. Male Wistar rats, stereotaxically implanted with intrahippocampal cannulas and electrodes, were injected with Tb II-I (2 µg/2 µL) via the intrahippocampal route. The behavior, electrographic activity and cellular integrity of the animals were analyzed and the intracerebral level of cytokines determined. Tb II-I injection induced seizures and damage in the hippocampus. These alterations were correlated with the changes in the level of the cytokines tumoral necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Therefore, the binding of Tb II-I to its target in the central nervous system may induce inflammation resulting in neuropathological and behavioral alterations.

ACS Style

Emidio Beraldo Neto; Douglas O. C. Mariano; Lucas A. Freitas; Ana L. C. Dorce; Adriana N. Martins; Daniel C. Pimenta; Fernanda C. V. Portaro; Daniela Cajado-Carvalho; Valquiria A. C. Dorce; Ana L. A. Nencioni. Tb II-I, a Fraction Isolated from Tityus bahiensis Scorpion Venom, Alters Cytokines’: Level and Induces Seizures When Intrahippocampally Injected in Rats. Toxins 2018, 10, 250 .

AMA Style

Emidio Beraldo Neto, Douglas O. C. Mariano, Lucas A. Freitas, Ana L. C. Dorce, Adriana N. Martins, Daniel C. Pimenta, Fernanda C. V. Portaro, Daniela Cajado-Carvalho, Valquiria A. C. Dorce, Ana L. A. Nencioni. Tb II-I, a Fraction Isolated from Tityus bahiensis Scorpion Venom, Alters Cytokines’: Level and Induces Seizures When Intrahippocampally Injected in Rats. Toxins. 2018; 10 (6):250.

Chicago/Turabian Style

Emidio Beraldo Neto; Douglas O. C. Mariano; Lucas A. Freitas; Ana L. C. Dorce; Adriana N. Martins; Daniel C. Pimenta; Fernanda C. V. Portaro; Daniela Cajado-Carvalho; Valquiria A. C. Dorce; Ana L. A. Nencioni. 2018. "Tb II-I, a Fraction Isolated from Tityus bahiensis Scorpion Venom, Alters Cytokines’: Level and Induces Seizures When Intrahippocampally Injected in Rats." Toxins 10, no. 6: 250.

Article
Published: 12 June 2018 in The Protein Journal
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A crucial step in scientific analysis can be sample preparation, and its importance increases in the same rate as the sensitivity of the following employed/desired analytical technique does. The need to analyze complex, viscous matrices is not new, and diverse approaches have been employed, with different success rates depending on the intended molecules. Solid-phase extraction, for example, has been successfully used in sample preparation for organic molecules and peptides. However, due to the usual methodological conditions, biologically active proteins are not successfully retrieved by this technique, resulting in a low rate of protein identification reported for the viscous amphibian skin secretion. Here we describe an ion-exchange batch processing sample preparation technique that allows viscous or adhesive materials (as some amphibian skin secretions) to be further processed by classical liquid chromatography approaches. According to our protocol, samples were allowed to equilibrate with a specific resin that was washed with appropriated buffers in order to yield the soluble protein fraction. In order to show the efficiency of our methodology, we have compared our results to classically prepared skin secretion, i.e., by means of filtration and centrifugation. After batch sample preparation, we were able to obtain reproductive resolved protein chromatographic profiles, as revealed by SDS-PAGE, and retrieve some biological activities, namely, hydrolases belonging to serine peptidase family. Not only that, but also the unbound fraction was rich in low molecular mass molecules, such as alkaloids and steroids, making this sample preparation technique also suitable for the enrichment of such molecules.

ACS Style

Douglas Mariano; Marcela Di Giacomo Messias; José Pedro Prezotto Neto; Patrick Spencer; Daniel C. Pimenta. Biochemical Analyses of Proteins from Duttaphrynus melanostictus (Bufo melanostictus) Skin Secretion: Soluble Protein Retrieval from a Viscous Matrix by Ion-Exchange Batch Sample Preparation. The Protein Journal 2018, 37, 380 -389.

AMA Style

Douglas Mariano, Marcela Di Giacomo Messias, José Pedro Prezotto Neto, Patrick Spencer, Daniel C. Pimenta. Biochemical Analyses of Proteins from Duttaphrynus melanostictus (Bufo melanostictus) Skin Secretion: Soluble Protein Retrieval from a Viscous Matrix by Ion-Exchange Batch Sample Preparation. The Protein Journal. 2018; 37 (4):380-389.

Chicago/Turabian Style

Douglas Mariano; Marcela Di Giacomo Messias; José Pedro Prezotto Neto; Patrick Spencer; Daniel C. Pimenta. 2018. "Biochemical Analyses of Proteins from Duttaphrynus melanostictus (Bufo melanostictus) Skin Secretion: Soluble Protein Retrieval from a Viscous Matrix by Ion-Exchange Batch Sample Preparation." The Protein Journal 37, no. 4: 380-389.

Journal article
Published: 24 October 2016 in Human & Experimental Toxicology
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Acquired immunodeficiency syndrome (AIDS) is a worldwide disease characterized by impairments of immune function. AIDS can be associated with oxidative stress (OS) that can be linked to selenium (Se) deficiency. Se is fundamental for the synthesis of selenoproteins, such as glutathione peroxidase and thioredoxin reductase. These enzymes catalyze the decomposition of reactive oxygen species and contribute to maintain equilibrium in cell redox status. Literature data indicate that organoselenium compounds, such as ebselen and diphenyl diselenide, have antioxidant properties in vitro and in vivo models associated with OS. Nevertheless, selenocompounds can also react and oxidize thiols groups, inducing toxicity in mammals. Here, we tested the potential cytotoxic and genotoxic properties of six analogs of the prototypal anti-HIV drug azidothymidine (AZT) containing Se (5′-Se-(phenyl)zidovudine; 5′-Se-(1,3,5-trimethylphenyl)zidovudine; 5′-Se-(1-naphtyl)zidovudine; 5′-Se-(4-chlorophenyl)zidovudine) (C4); 5′-Se-(4-methylphenyl)zidovudine (C5); and 5′-(4-methylbenzoselenoate)zidovudine). C5 increased the rate of dithiothreitol oxidation (thiol oxidase activity) and C2-C4 and C6 (at 100 µM) increased DNA damage index (DI) in human leukocytes. Moreover, C5 (200 µM) decreased human leukocyte viability to about 50%. Taken together, these results indicated the low in vitro toxicity in human leukocytes of some Se-containing analogs of AZT.

ACS Style

Douglas Mariano; Diego Souza; D F Meinerz; J Allebrandt; Andreza Fabro de Bem; W Hassan; Oscar Endrigo Dorneles Rodrigues; Jbt Da Rocha. The potential toxicological insights about the anti-HIV drug azidothymidine-derived monoselenides in human leukocytes: Toxicological insights of new selenium-azidothymidine analogs. Human & Experimental Toxicology 2016, 36, 910 -918.

AMA Style

Douglas Mariano, Diego Souza, D F Meinerz, J Allebrandt, Andreza Fabro de Bem, W Hassan, Oscar Endrigo Dorneles Rodrigues, Jbt Da Rocha. The potential toxicological insights about the anti-HIV drug azidothymidine-derived monoselenides in human leukocytes: Toxicological insights of new selenium-azidothymidine analogs. Human & Experimental Toxicology. 2016; 36 (9):910-918.

Chicago/Turabian Style

Douglas Mariano; Diego Souza; D F Meinerz; J Allebrandt; Andreza Fabro de Bem; W Hassan; Oscar Endrigo Dorneles Rodrigues; Jbt Da Rocha. 2016. "The potential toxicological insights about the anti-HIV drug azidothymidine-derived monoselenides in human leukocytes: Toxicological insights of new selenium-azidothymidine analogs." Human & Experimental Toxicology 36, no. 9: 910-918.

Research article
Published: 08 April 2015 in Journal of Medicinal Chemistry
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In this article we present the synthesis, characterization, and in vitro biological and biochemical activities of new chalcogenozidovudine derivatives as antioxidant (inhibition of TBARS in brain membranes and thiol peroxidase-like activity) as well as antitumoral agents in bladder carcinoma 5637. A prominent response was obtained for the selected chalcogenonucleosides, showing effective antioxidant and antitumoral activities.

ACS Style

Diego Souza; Douglas Mariano; Fernanda Nedel; Eduarda Schultze; Vinicius Farias Campos; Fabiana Kommling Seixas; Rafael S. Da Silva; Taiana S. Munchen; Vinicius Ilha; Luciano Dornelles; Antonio Braga; Joao Batista Teixeira da Rocha; Tiago Collares; Oscar E. D. Rodrigues. New Organochalcogen Multitarget Drug: Synthesis and Antioxidant and Antitumoral Activities of Chalcogenozidovudine Derivatives. Journal of Medicinal Chemistry 2015, 58, 3329 -3339.

AMA Style

Diego Souza, Douglas Mariano, Fernanda Nedel, Eduarda Schultze, Vinicius Farias Campos, Fabiana Kommling Seixas, Rafael S. Da Silva, Taiana S. Munchen, Vinicius Ilha, Luciano Dornelles, Antonio Braga, Joao Batista Teixeira da Rocha, Tiago Collares, Oscar E. D. Rodrigues. New Organochalcogen Multitarget Drug: Synthesis and Antioxidant and Antitumoral Activities of Chalcogenozidovudine Derivatives. Journal of Medicinal Chemistry. 2015; 58 (8):3329-3339.

Chicago/Turabian Style

Diego Souza; Douglas Mariano; Fernanda Nedel; Eduarda Schultze; Vinicius Farias Campos; Fabiana Kommling Seixas; Rafael S. Da Silva; Taiana S. Munchen; Vinicius Ilha; Luciano Dornelles; Antonio Braga; Joao Batista Teixeira da Rocha; Tiago Collares; Oscar E. D. Rodrigues. 2015. "New Organochalcogen Multitarget Drug: Synthesis and Antioxidant and Antitumoral Activities of Chalcogenozidovudine Derivatives." Journal of Medicinal Chemistry 58, no. 8: 3329-3339.

Journal article
Published: 01 January 2015 in Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology
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The presence of peptides has been identified in all African pipid genera; nevertheless, little is known about skin secretion of South American frog genus Pipa. Skin secretion from captive and wild Pipa carvalhoi were obtained in the presence or absence of norepinephrine stimulation. The <10 kDa fraction was analyzed by liquid chromatography and mass spectrometry, searching for peptides. Chromatographic profiles show the presence of a major component in this secretion, regardless of the stimulation method (norepinephrine or mechanical stimulation) and the origin of the animal (captivity or wild), as well as in the absence of any stimulus. The general mass distribution profile in P. carvalhoi skin secretion shows numerous components below 800 Da. Moreover, no peptide could be identified, regardless of the chromatographic approach. The major component was purified and identified as kynurenic acid, an L-tryptophan derivative. P. carvalhoi does not secrete peptides as toxins in its skin. In addition, we here report that kynurenic acid is the main component of P. carvalhoi skin secretion. Although no biological activity was associated with kynurenic acid, we propose that this molecule is a pheromone that signals the presence of a co-specific in the shady environment in which this animal lives. In this study we demonstrate the absence of peptidic toxins in the skin secretion of P. carvalhoi, a break of paradigm in the pipid family.

ACS Style

Douglas Oscar Ceolin Mariano; Lydia Fumiko Yamaguchi; Carlos Jared; Marta Maria Antoniazzi; Juliana Mozer Sciani; Massuo Jorge Kato; Daniel Carvalho Pimenta. Pipa carvalhoi skin secretion profiling: Absence of peptides and identification of kynurenic acid as the major constitutive component. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2015, 167, 1 -6.

AMA Style

Douglas Oscar Ceolin Mariano, Lydia Fumiko Yamaguchi, Carlos Jared, Marta Maria Antoniazzi, Juliana Mozer Sciani, Massuo Jorge Kato, Daniel Carvalho Pimenta. Pipa carvalhoi skin secretion profiling: Absence of peptides and identification of kynurenic acid as the major constitutive component. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology. 2015; 167 ():1-6.

Chicago/Turabian Style

Douglas Oscar Ceolin Mariano; Lydia Fumiko Yamaguchi; Carlos Jared; Marta Maria Antoniazzi; Juliana Mozer Sciani; Massuo Jorge Kato; Daniel Carvalho Pimenta. 2015. "Pipa carvalhoi skin secretion profiling: Absence of peptides and identification of kynurenic acid as the major constitutive component." Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 167, no. : 1-6.

Randomized controlled trial
Published: 01 April 2014 in Nutrition
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The aim of this study was to investigate the effect of a single dose of Brazil nuts on the inflammatory markers of healthy individuals. A randomized crossover study was conducted with 10 healthy individuals (mean age 24.7 ± 3.4 y). Each individual was tested four times regarding intake of different portions of Brazil nuts: 0, 5, 20 and 50 g. At each testing period, peripheral blood was collected before and at 1, 3, 6, 9, 24, and 48 h after intake of nuts, as well as at 5 and 30 d after intake of various Brazil nut portions. Blood samples were tested for high-sensitivity to C-reactive protein, interleukin (IL)-1, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, aspartate and alanine aminotransferases, albumin, total protein, alkaline phosphatase, gamma-glutamyltransferase, urea, and creatinine. Consumption of nuts did not affect biochemical parameters for liver and kidney function, indicating absence of hepatic and renal toxicity. A single intake of Brazil nuts (20 or 50 g) caused a significant decrease in serum IL-1, IL-6, TNF-α, and IFN-γ levels (P < 0.05), whereas serum levels of IL-10 were significantly increased (P < 0.05). The results indicate a long-term decrease in inflammatory markers after a single intake of large portions of Brazil nuts in healthy volunteers. Therefore, the long-term effect of regular Brazil nut consumption on inflammatory markers should be better investigated.

ACS Style

Elisângela Colpo; Carlos Dalton D.A. Vilanova; Luiz Gustavo B. Reetz; Marta M.M.F. Duarte; Iria Luiza G. Farias; Daiane F. Meinerz; Douglas O.C. Mariano; Raquel G. Vendrusculo; Aline A. Boligon; Cristiane L. Dalla Corte; Roger Wagner; Margareth L. Athayde; João Batista T. da Rocha. Brazilian nut consumption by healthy volunteers improves inflammatory parameters. Nutrition 2014, 30, 459 -465.

AMA Style

Elisângela Colpo, Carlos Dalton D.A. Vilanova, Luiz Gustavo B. Reetz, Marta M.M.F. Duarte, Iria Luiza G. Farias, Daiane F. Meinerz, Douglas O.C. Mariano, Raquel G. Vendrusculo, Aline A. Boligon, Cristiane L. Dalla Corte, Roger Wagner, Margareth L. Athayde, João Batista T. da Rocha. Brazilian nut consumption by healthy volunteers improves inflammatory parameters. Nutrition. 2014; 30 (4):459-465.

Chicago/Turabian Style

Elisângela Colpo; Carlos Dalton D.A. Vilanova; Luiz Gustavo B. Reetz; Marta M.M.F. Duarte; Iria Luiza G. Farias; Daiane F. Meinerz; Douglas O.C. Mariano; Raquel G. Vendrusculo; Aline A. Boligon; Cristiane L. Dalla Corte; Roger Wagner; Margareth L. Athayde; João Batista T. da Rocha. 2014. "Brazilian nut consumption by healthy volunteers improves inflammatory parameters." Nutrition 30, no. 4: 459-465.

Journal article
Published: 18 March 2014 in PeerJ
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Organoselenium compounds have been pointed out as therapeutic agents. In contrast, the potential therapeutic aspects of tellurides have not yet been demonstrated. The present study evaluated the comparative toxicological effects of diphenyl diselenide (PhSe)2 and diphenyl ditelluride (PhTe)2 in mice after in vivo administration. Genotoxicity (as determined by comet assay) and mutagenicicity were used as end-points of toxicity. Subcutaneous administration of high doses of (PhSe)2 or (PhTe)2 (500 µmol/kg) caused distinct genotoxicity in mice. (PhSe)2 significantly decreased the DNA damage index after 48 and 96 h of its injection (p < 0.05). In contrast, (PhTe) caused a significant increase in DNA damage (p < 0.05) after 48 and 96 h of intoxication. (PhSe)2 did not cause mutagenicity but (PhTe)2 increased the micronuclei frequency, indicating its mutagenic potential. The present study demonstrated that acute in vivo exposure to ditelluride caused genotoxicity in mice, which may be associated with pro-oxidant effects of diphenyl ditelluride. In addition, the use of this compound and possibly other related tellurides must be carefully controlled

ACS Style

Daiane Francine Meinerz; Josiane Allebrandt; Douglas Mariano; Emily P. Waczuk; Félix Soares; Waseem Hassan; João Batista T. Rocha. Differential genotoxicity of diphenyl diselenide (PhSe)2and diphenyl ditelluride (PhTe)2. PeerJ 2014, 2, e290 .

AMA Style

Daiane Francine Meinerz, Josiane Allebrandt, Douglas Mariano, Emily P. Waczuk, Félix Soares, Waseem Hassan, João Batista T. Rocha. Differential genotoxicity of diphenyl diselenide (PhSe)2and diphenyl ditelluride (PhTe)2. PeerJ. 2014; 2 ():e290.

Chicago/Turabian Style

Daiane Francine Meinerz; Josiane Allebrandt; Douglas Mariano; Emily P. Waczuk; Félix Soares; Waseem Hassan; João Batista T. Rocha. 2014. "Differential genotoxicity of diphenyl diselenide (PhSe)2and diphenyl ditelluride (PhTe)2." PeerJ 2, no. : e290.

Comparative study
Published: 21 November 2013 in BioMed Research International
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Organochalcogens, particularly ebselen, have been used in experimental and clinical trials with borderline efficacy. (PhSe)2and (PhTe)2are the simplest of the diaryl dichalcogenides and share with ebselen pharmacological properties. In view of the concerns with the use of mammals in studies and the great number of new organochalcogens with potential pharmacological properties that have been synthesized, it becomes important to develop screening protocols to select compounds that are worth to be testedin vivo. This study investigated the possible use of isolated human white cells as a preliminary model to test organochalcogen toxicity. Human leucocytes were exposed to 5–50 μM of ebselen, (PhSe)2, or (PhTe)2. All compounds were cytotoxic (Trypan’s Blue exclusion) at the highest concentration tested, and Ebselen was the most toxic. Ebselen and (PhSe)2were genotoxic (Comet Assay) only at 50 μM, and (PhTe)2at 5–50 μM. Here, the acute cytotoxicity did not correspond within vivotoxicity of the compounds. But the genotoxicity was in the same order of thein vivotoxicity to mice. These results indicate thatin vitrogenotoxicity in white blood cells should be considered as an early step in the investigation of potential toxicity of organochalcogens.

ACS Style

Diones Bueno; Daiane Francine Meinerz; Josiane Allebrandt; Emily Pansera Waczuk; Danúbia Bonfanti Dos Santos; Douglas Mariano; João Batista Teixeira Rocha. Cytotoxicity and Genotoxicity Evaluation of Organochalcogens in Human Leucocytes: A Comparative Study between Ebselen, Diphenyl Diselenide, and Diphenyl Ditelluride. BioMed Research International 2013, 2013, 1 -6.

AMA Style

Diones Bueno, Daiane Francine Meinerz, Josiane Allebrandt, Emily Pansera Waczuk, Danúbia Bonfanti Dos Santos, Douglas Mariano, João Batista Teixeira Rocha. Cytotoxicity and Genotoxicity Evaluation of Organochalcogens in Human Leucocytes: A Comparative Study between Ebselen, Diphenyl Diselenide, and Diphenyl Ditelluride. BioMed Research International. 2013; 2013 ():1-6.

Chicago/Turabian Style

Diones Bueno; Daiane Francine Meinerz; Josiane Allebrandt; Emily Pansera Waczuk; Danúbia Bonfanti Dos Santos; Douglas Mariano; João Batista Teixeira Rocha. 2013. "Cytotoxicity and Genotoxicity Evaluation of Organochalcogens in Human Leucocytes: A Comparative Study between Ebselen, Diphenyl Diselenide, and Diphenyl Ditelluride." BioMed Research International 2013, no. : 1-6.

Journal article
Published: 24 April 2013 in SpringerPlus
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The organic tellurium compound (S)-dimethyl 2-(3-(phenyltellanyl) propanamide) succinate (TeAsp) exhibits thiol-peroxidase activity that could potentially offer protection against oxidative stress. However, data from the literature show that tellurium is a toxic agent to rodents. In order to mitigate such toxicity, N-acetylcysteine (NAC) was administered in parallel with TeAsp during 10 days. Mice were separated into four groups receiving daily injections of (A) vehicle (PBS 2.5 ml/kg, i.p. and DMSO 1 ml/kg, s.c.), (B) NAC (100 mg/kg, i.p. and DMSO s.c.), (C) PBS i.p. and TeAsp (92.5 μmol/kg, s.c), or (D) NAC plus TeAsp. TeAsp treatment started on the fourth day. Vehicle or NAC-treated animals showed an increase in body weight whereas TeAsp caused a significant reduction. Contrary to expected, NAC co-administration potentiated the toxic effect of TeAsp, causing a decrease in body weight. Vehicle, NAC or TeAsp did not affect the exploratory and motor activity in the open-field test at the end of the treatment, while the combination of NAC and TeAsp produced a significant decrease in these parameters. No DNA damage or alterations in cell viability were observed in leukocytes of treated animals. Treatments produced no or minor effects on the activities of antioxidant enzymes catalase, glutathione peroxidase and glutathione reductase, whereas the activity of the thioredoxin reductase was decreased in the brain and increased the liver of the animals in the groups receiving TeAsp or TeAsp plus NAC. In conclusion, the toxicity of TeAsp was potentiated by NAC and oxidative stress appears to play a central role in this process.

ACS Style

Daiane F Meinerz; Bruna Comparsi; Josiane Allebrandt; Douglas Oscar Ceolin Mariano; Danúbia B Dos Santos; Ana Paula Pegoraro Zemolin; Marcelo Farina; Luiz Alcir Dafre; João B T Rocha; Thais Posser; Jeferson Luis Franco. Sub-acute administration of (S)-dimethyl 2-(3-(phenyltellanyl) propanamido) succinate induces toxicity and oxidative stress in mice: unexpected effects of N-acetylcysteine. SpringerPlus 2013, 2, 182 .

AMA Style

Daiane F Meinerz, Bruna Comparsi, Josiane Allebrandt, Douglas Oscar Ceolin Mariano, Danúbia B Dos Santos, Ana Paula Pegoraro Zemolin, Marcelo Farina, Luiz Alcir Dafre, João B T Rocha, Thais Posser, Jeferson Luis Franco. Sub-acute administration of (S)-dimethyl 2-(3-(phenyltellanyl) propanamido) succinate induces toxicity and oxidative stress in mice: unexpected effects of N-acetylcysteine. SpringerPlus. 2013; 2 (1):182.

Chicago/Turabian Style

Daiane F Meinerz; Bruna Comparsi; Josiane Allebrandt; Douglas Oscar Ceolin Mariano; Danúbia B Dos Santos; Ana Paula Pegoraro Zemolin; Marcelo Farina; Luiz Alcir Dafre; João B T Rocha; Thais Posser; Jeferson Luis Franco. 2013. "Sub-acute administration of (S)-dimethyl 2-(3-(phenyltellanyl) propanamido) succinate induces toxicity and oxidative stress in mice: unexpected effects of N-acetylcysteine." SpringerPlus 2, no. 1: 182.

Journal article
Published: 01 December 2012 in Toxicology
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We evaluated the activity and expression of antioxidant enzymes in the cerebellum and cortex of Swiss adult male mice exposed to methylmercury (MeHg) in drinking water (40mg/L) during 21 days. The activity of glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were determined spectrophotometrically. The expression (protein levels) of GPx1 and GPx4 isoforms, TrxR1 as well as heat shock protein 70 (HSP70) were evaluated using specific antibodies and normalized by actin levels. The exposure of mice to MeHg caused a significant impairment in locomotors performance in the open field test (crossings and rearing). This result was followed by a significant reduction of GPx and TrxR activities in the cerebellum and cortex when compared to untreated animals. We also observed a substantial decrease in GPx1, GPx4 and TrxR1 protein levels in the cerebellum, while in the cerebral cortex, only GPx4 and TrxR1 were decreased after MeHg treatment. The activities of the antioxidant enzymes GR, GST, CAT and SOD were increased in the cerebellum after MeHg administration to mice. In contrast, only CAT was increased in the cerebral cortex of MeHg-treated animals. The expression of HSP70 was up-regulated only in the cerebellum where MeHg-exposed mice showed a significant increase in the immunocontent of HSP70 when compared to controls. This is the first report showing a role for GPx4 in the neurotoxicity induced by MeHg in vivo. In addition, our data indicates that the selenoproteins GPx and TrxR as main targets during MeHg exposure, which may be considered in biomarker studies.

ACS Style

A.P.P. Zemolin; D.F. Meinerz; Mariane de Paula; Douglas Mariano; Joao Batista Teixeira da Rocha; A.B. Pereira; T. Posser; Jeferson Luis Franco. Evidences for a role of glutathione peroxidase 4 (GPx4) in methylmercury induced neurotoxicity in vivo. Toxicology 2012, 302, 60 -67.

AMA Style

A.P.P. Zemolin, D.F. Meinerz, Mariane de Paula, Douglas Mariano, Joao Batista Teixeira da Rocha, A.B. Pereira, T. Posser, Jeferson Luis Franco. Evidences for a role of glutathione peroxidase 4 (GPx4) in methylmercury induced neurotoxicity in vivo. Toxicology. 2012; 302 (1):60-67.

Chicago/Turabian Style

A.P.P. Zemolin; D.F. Meinerz; Mariane de Paula; Douglas Mariano; Joao Batista Teixeira da Rocha; A.B. Pereira; T. Posser; Jeferson Luis Franco. 2012. "Evidences for a role of glutathione peroxidase 4 (GPx4) in methylmercury induced neurotoxicity in vivo." Toxicology 302, no. 1: 60-67.