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Dr. Daniel Pimenta
Butantan Institute

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0 Amphibians
0 Chromatography
0 Enzyme Activity
0 Marine Biology
0 Mass Spectrometry

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Peptides
Mass Spectrometry
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Journal article
Published: 30 June 2021 in Toxins
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Osteoclasts (OCs) are important for bone maintenance, calcium balance, and tissue regeneration regulation and are involved in different inflammatory diseases. Our study aimed to evaluate the effect of Bothrops moojeni’s venom and its low and high molecular mass (HMM and LMM) fractions on human peripheral blood mononuclear cell (PBMC)-derived OCs’ in vitro differentiation. Bothrops moojeni, a Brazilian lanced-head viper, presents a rich but not well-explored, venom composition. This venom is a potent inducer of inflammation, which can be used as a tool to investigate the inflammatory process. Human PBMCs were isolated and induced to OC differentiation following routine protocol. On the fourth day of differentiation, the venom was added at different concentrations (5, 0.5, and 0.05 µg/mL). We observed a significant reduction of TRAP+ (tartrate-resistant acid phosphatase) OCs at the concentration of 5 µg/mL. We evaluated the F-actin-rich OCs structure’s integrity; disruption of its integrity reflects bone adsorption capacity. F-actin rings phalloidin staining demonstrated that venom provoked their disruption in treated OCs. HMM, fraction reduces TRAP+ OCs at a concentration of 5 µg/mL and LMM fraction at 1 µg/mL, respectively. Our results indicate morphological changes that the venom induced cause in OCs. We analyzed the pattern of soluble proteins found in the conditioned cell culture medium OCs treated with venom and its fractions using mass spectrometry (LC-MS/IT-Tof). The proteomic analyses indicate the possible pathways and molecular mechanisms involved in OC reduction after the treatment.

ACS Style

Fernanda D’Amélio; Hugo Vigerelli; Álvaro Prieto-Da-Silva; Eduardo Osório Frare; Isabel Batista; Daniel Pimenta; Irina Kerkis. Bothrops moojeni Venom and Its Components Strongly Affect Osteoclasts’ Maturation and Protein Patterns. Toxins 2021, 13, 459 .

AMA Style

Fernanda D’Amélio, Hugo Vigerelli, Álvaro Prieto-Da-Silva, Eduardo Osório Frare, Isabel Batista, Daniel Pimenta, Irina Kerkis. Bothrops moojeni Venom and Its Components Strongly Affect Osteoclasts’ Maturation and Protein Patterns. Toxins. 2021; 13 (7):459.

Chicago/Turabian Style

Fernanda D’Amélio; Hugo Vigerelli; Álvaro Prieto-Da-Silva; Eduardo Osório Frare; Isabel Batista; Daniel Pimenta; Irina Kerkis. 2021. "Bothrops moojeni Venom and Its Components Strongly Affect Osteoclasts’ Maturation and Protein Patterns." Toxins 13, no. 7: 459.

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.

Preprint content
Published: 03 January 2021
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Safety, optimal minimum dose, and, preliminary effectiveness of a new generation Africanized honeybees (Apis mellifera) antivenom (AAV) were evaluated. A phase I/II, multicenter, non- randomized, single-arm clinical trial involving 20 participants showing multiple stings were studied. Participants have received either 2 to 10 vials of AAV based on the stings number together with a predefined adjuvant, symptomatic, and complementary treatment schedule. The primary safety endpoint was the presence of early adverse reactions within the first 24 hours after treatment. Preliminary efficacy through clinical evolution, including laboratory tests, was assessed at baseline and over the following four weeks. ELISA assays and mass spectrometry estimated the venom pharmacokinetics before, during, and after treatment. Twenty adult participants, 13 (65%) males, and 7 (35%) females, with a median age of 44 years and a mean body surface of 1.92 m2 (median = 1.93 m2) were recruited. The median number of stings was 52.5 ranging from 7 to more than 2,000. Envenoming severity was classified as 80% mild, 15% moderate, and 5% severe. According to the protocol, 16 (80%) participants received two AAV vials, 3 (15%) six vials, and one (5%) 10 vials. There was no discontinuation of the treatment due to acute adverse events and there were no late adverse reactions. Two patients showed mild adverse events with only transient itchy skin and erythroderma. All participants completed the infusion within two hours and there was no loss of follow-up after discharge. ELISA assays showed venom concentrations varying between 0.25 ng/mL and 1.479 ng/mL prior to treatment. Venom levels decreased in all cases during the hospitalization period. Surprisingly, in nine cases (45%), despite clinical recovery and without symptoms, the venom levels increased again during outpatient care 10 days after discharge. Mass spectrometry showed melittin in eight participants 30 days after the treatment. Considering the promising safety results of the investigational product for the treatment of massive Africanized honeybee attacks, added to efficacy in clinical improvement and immediate decrease in blood venom level, the AAV has shown to be safe for human use. Trial registration Universal Trial Number (UTN): U1111-1160-7011, Register Number: RBR-3fthf8 (http://www.ensaiosclinicos.gov.br/rg/RBR-3fthf8/).

ACS Style

Alexandre Naime Barbosa; Rui Seabra Ferreira; Francilene Capel Tavares de Carvalho; Fabiana Schuelter-Trevisol; Mônica Bannwart Mendes; Bruna Cavecci Mendonça; José Nixon Batista; Daisson José Trevisol; Leslie Boyer; Jean-Philippe Chippaux; Natália Bronzatto Medolago; Claudia Vilalva Cassaro; Márcia Tonin Rigotto Carneiro; Ana Paola Piloto de Oliveira; Daniel Carvalho Pimenta; Luís Eduardo Ribeiro da Cunha; Lucilene Delazari dos Santos; Benedito Barraviera. A phase I/II trial to treat massive Africanized honeybee (Apis mellifera) stings using the new apilic antivenom. 2021, 1 .

AMA Style

Alexandre Naime Barbosa, Rui Seabra Ferreira, Francilene Capel Tavares de Carvalho, Fabiana Schuelter-Trevisol, Mônica Bannwart Mendes, Bruna Cavecci Mendonça, José Nixon Batista, Daisson José Trevisol, Leslie Boyer, Jean-Philippe Chippaux, Natália Bronzatto Medolago, Claudia Vilalva Cassaro, Márcia Tonin Rigotto Carneiro, Ana Paola Piloto de Oliveira, Daniel Carvalho Pimenta, Luís Eduardo Ribeiro da Cunha, Lucilene Delazari dos Santos, Benedito Barraviera. A phase I/II trial to treat massive Africanized honeybee (Apis mellifera) stings using the new apilic antivenom. . 2021; ():1.

Chicago/Turabian Style

Alexandre Naime Barbosa; Rui Seabra Ferreira; Francilene Capel Tavares de Carvalho; Fabiana Schuelter-Trevisol; Mônica Bannwart Mendes; Bruna Cavecci Mendonça; José Nixon Batista; Daisson José Trevisol; Leslie Boyer; Jean-Philippe Chippaux; Natália Bronzatto Medolago; Claudia Vilalva Cassaro; Márcia Tonin Rigotto Carneiro; Ana Paola Piloto de Oliveira; Daniel Carvalho Pimenta; Luís Eduardo Ribeiro da Cunha; Lucilene Delazari dos Santos; Benedito Barraviera. 2021. "A phase I/II trial to treat massive Africanized honeybee (Apis mellifera) stings using the new apilic antivenom." , no. : 1.

Research
Published: 01 January 2021 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Amphibians inhabit the terrestrial environment, a conquest achieved after several evolutionary steps, which were still insufficient to make them completely independent of the aquatic environment. These processes gave rise to many morphological and physiological changes, making their skin (and cutaneous secretion) rich in bioactive molecules. Among the tree frogs, the secretion is composed mainly of peptides; but alkaloids, proteins and steroids can also be found depending on the species. The most known class of biologically active molecules is the antimicrobial peptides (AMPs) that act against bacteria, fungi and protozoans. Although these molecules are well-studied among the hylids, AMPs ontogeny remains unknown. Therefore, we performed peptidomic and proteomic analyses of Pithecopus nordestinus (formerly Phyllomedusa nordestina) in order to evaluate the peptide content in post-metamorphosed juveniles and adult individuals. Cutaneous secretion of both life stages of individuals was obtained and analyzed by LC-MS/MS after reduction and alkylation of disulfide bonds or reduction, alkylation and hydrolysis by trypsin. Differences in the TIC profile of juveniles and adults in both treatments were observed. Moreover, the proteomic data revealed known proteins and peptides, with slight differences in the composition, according to the life stage and the treatment. AMPs were identified, and bradykinin-potentiating peptides were observed in trypsin-treated samples, which suggests a protein source of such peptide (cryptide). In general, skin secretion contents were similar between juveniles and adults, varying in quantity, indicating that the different stages of life are reflected in the number of molecules and not on their diversity.

ACS Style

Douglas O. Mariano; Juliana M. Sciani; Marta M. Antoniazzi; Carlos Jared; Katia Conceição; Daniel C. Pimenta. Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus. Journal of Venomous Animals and Toxins including Tropical Diseases 2021, 27, 1 .

AMA Style

Douglas O. Mariano, Juliana M. Sciani, Marta M. Antoniazzi, Carlos Jared, Katia Conceição, Daniel C. Pimenta. Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus. Journal of Venomous Animals and Toxins including Tropical Diseases. 2021; 27 ():1.

Chicago/Turabian Style

Douglas O. Mariano; Juliana M. Sciani; Marta M. Antoniazzi; Carlos Jared; Katia Conceição; Daniel C. Pimenta. 2021. "Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus." Journal of Venomous Animals and Toxins including Tropical Diseases 27, no. : 1.

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.

Conference abstract
Published: 01 April 2020 in Toxicon
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ACS Style

Luciano Fusco; Emídio Neto; Aleff Francisco; Jorge Alfonso; Claudia Siqueira; Laura Leiva; Daniel Pimenta; Andreimar Soares. Snake venomics of Crotalus durissus terrificus from Argentina. Toxicon 2020, 177, S51 -S52.

AMA Style

Luciano Fusco, Emídio Neto, Aleff Francisco, Jorge Alfonso, Claudia Siqueira, Laura Leiva, Daniel Pimenta, Andreimar Soares. Snake venomics of Crotalus durissus terrificus from Argentina. Toxicon. 2020; 177 ():S51-S52.

Chicago/Turabian Style

Luciano Fusco; Emídio Neto; Aleff Francisco; Jorge Alfonso; Claudia Siqueira; Laura Leiva; Daniel Pimenta; Andreimar Soares. 2020. "Snake venomics of Crotalus durissus terrificus from Argentina." Toxicon 177, no. : S51-S52.

Journal article
Published: 21 January 2020 in Toxins
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Here, we report the neurotoxic effects aroused by the intracerebral injection (in rats) of Tb1, which is a neurotoxin isolated from Tityus bahiensis scorpion venom. Biochemical analyses have demonstrated that this toxin is similar to the gamma toxin from T. serrulatus, which is a β-scorpion toxin that acts on sodium channels, causing the activation process to occur at more hyperpolarized membrane voltages. Male Wistar rats were stereotaxically implanted with intrahippocampal electrodes and cannulas for electroencephalographic recording and the evaluation of amino acid neurotransmitters levels. Treated animals displayed behavioral and electroencephalographic alterations similar to epileptiform activities, such as myoclonus, wet dog shakes, convulsion, strong discharges, neuronal loss, and increased intracerebral levels of glutamate. Scorpion toxins are important pharmacological tools that are widely employed in ion channel dysregulation studies. The current work contributes to the understanding of channelopathies, particularly epilepsy, which may originate, among other events, from dysfunctional sodium channels, which are the main target of the Tb1 toxin.

ACS Style

Emidio Beraldo Neto; Lucas Alves De Freitas; Daniel Carvalho Pimenta; Ivo Lebrun; Ana L. A. Nencioni. Tb1, a Neurotoxin from Tityus bahiensis Scorpion Venom, Induces Epileptic Seizures by Increasing Glutamate Release. Toxins 2020, 12, 65 .

AMA Style

Emidio Beraldo Neto, Lucas Alves De Freitas, Daniel Carvalho Pimenta, Ivo Lebrun, Ana L. A. Nencioni. Tb1, a Neurotoxin from Tityus bahiensis Scorpion Venom, Induces Epileptic Seizures by Increasing Glutamate Release. Toxins. 2020; 12 (2):65.

Chicago/Turabian Style

Emidio Beraldo Neto; Lucas Alves De Freitas; Daniel Carvalho Pimenta; Ivo Lebrun; Ana L. A. Nencioni. 2020. "Tb1, a Neurotoxin from Tityus bahiensis Scorpion Venom, Induces Epileptic Seizures by Increasing Glutamate Release." Toxins 12, no. 2: 65.

Research
Published: 01 January 2020 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Between 40,000-70,000 people die yearly of rabies, an incurable disease. Besides post-bite vaccination, no treatment is available for it. First, virus dilution for antiviral effects in mice was determined. Then, animals were treated as follows: control (NaCl 250 µL/animal/day); bufotenine (0.63, 1.05 and 2.1 mg in 250 µL of NaCl/animal/day); rabies (10-6,82CVS dilution); and test (10-6,82 CVS dilution and bufotenine, in the above-mentioned doses). Animals were observed daily for 21 days or until the 3rd stage of rabies infection. Twitch-tension and liposome studies were applied to understand the possible interaction of bufotenine with receptors, particularly acetylcholine. Bufotenine was able to increase the survival rate of intracerebrally virus-infected mice from 15 to 40%. Bufotenine did not seem to interfere with the acetylcholine response in the skeletal muscle, indicating that its mechanism of action is not blocking the virus entrance due to nAChR antagonism. By analyzing liposomes, we could observe that bufotenine did not passively penetrates cell membranes, indicating the necessity of complementary structures to cell penetration. Bufotenine is a promising candidate for drug development. After further chemical modification, it might be possible to dissociate minor side effects, increase efficiency, efficacy and pharmacokinetics, yielding a true anti-rabies drug.

ACS Style

Hugo Vigerelli; Juliana Sciani; Patricia M. C. Pereira; Aline A. Lavezo; Andrea C. R. Silva; Rita Collaço; Thalita Rocha; Thais C. Bueno; Daniel C. Pimenta. Bufotenine, a tryptophan-derived alkaloid, suppresses the symptoms and increases the survival rate of rabies-infected mice: the development of a pharmacological approach for rabies treatment. Journal of Venomous Animals and Toxins including Tropical Diseases 2020, 26, e20190050 .

AMA Style

Hugo Vigerelli, Juliana Sciani, Patricia M. C. Pereira, Aline A. Lavezo, Andrea C. R. Silva, Rita Collaço, Thalita Rocha, Thais C. Bueno, Daniel C. Pimenta. Bufotenine, a tryptophan-derived alkaloid, suppresses the symptoms and increases the survival rate of rabies-infected mice: the development of a pharmacological approach for rabies treatment. Journal of Venomous Animals and Toxins including Tropical Diseases. 2020; 26 ():e20190050.

Chicago/Turabian Style

Hugo Vigerelli; Juliana Sciani; Patricia M. C. Pereira; Aline A. Lavezo; Andrea C. R. Silva; Rita Collaço; Thalita Rocha; Thais C. Bueno; Daniel C. Pimenta. 2020. "Bufotenine, a tryptophan-derived alkaloid, suppresses the symptoms and increases the survival rate of rabies-infected mice: the development of a pharmacological approach for rabies treatment." Journal of Venomous Animals and Toxins including Tropical Diseases 26, no. : e20190050.

Research
Published: 01 January 2020 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.

ACS Style

Roberto Tadashi Kodama; Alexandre Kazuo Kuniyoshi; Cristiane Castilho Fernandes Da Silva; Daniela Cajado-Carvalho; Bruno Duzzi; Douglas Ceolin Mariano; Daniel C. Pimenta; Rafael Borges; Wilmar Dias Da Silva; Fernanda Calheta Vieira Portaro. A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases. Journal of Venomous Animals and Toxins including Tropical Diseases 2020, 26, 1 .

AMA Style

Roberto Tadashi Kodama, Alexandre Kazuo Kuniyoshi, Cristiane Castilho Fernandes Da Silva, Daniela Cajado-Carvalho, Bruno Duzzi, Douglas Ceolin Mariano, Daniel C. Pimenta, Rafael Borges, Wilmar Dias Da Silva, Fernanda Calheta Vieira Portaro. A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases. Journal of Venomous Animals and Toxins including Tropical Diseases. 2020; 26 ():1.

Chicago/Turabian Style

Roberto Tadashi Kodama; Alexandre Kazuo Kuniyoshi; Cristiane Castilho Fernandes Da Silva; Daniela Cajado-Carvalho; Bruno Duzzi; Douglas Ceolin Mariano; Daniel C. Pimenta; Rafael Borges; Wilmar Dias Da Silva; Fernanda Calheta Vieira Portaro. 2020. "A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases." Journal of Venomous Animals and Toxins including Tropical Diseases 26, no. : 1.

Research
Published: 01 January 2020 in Journal of Venomous Animals and Toxins including Tropical Diseases
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Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs.

ACS Style

Luciano S. Fusco; Rodrigo Cajade; Jose M. Piñeiro; Ana M. Torres; Igor R. F. Da Silva; Stephen Hyslop; Laura C. Leiva; Daniel C. Pimenta; Soledad Bustillo. Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae). Journal of Venomous Animals and Toxins including Tropical Diseases 2020, 26, e20190078 .

AMA Style

Luciano S. Fusco, Rodrigo Cajade, Jose M. Piñeiro, Ana M. Torres, Igor R. F. Da Silva, Stephen Hyslop, Laura C. Leiva, Daniel C. Pimenta, Soledad Bustillo. Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae). Journal of Venomous Animals and Toxins including Tropical Diseases. 2020; 26 ():e20190078.

Chicago/Turabian Style

Luciano S. Fusco; Rodrigo Cajade; Jose M. Piñeiro; Ana M. Torres; Igor R. F. Da Silva; Stephen Hyslop; Laura C. Leiva; Daniel C. Pimenta; Soledad Bustillo. 2020. "Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)." Journal of Venomous Animals and Toxins including Tropical Diseases 26, no. : e20190078.

Journal article
Published: 06 December 2019 in Scientific Reports
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Tetrodotoxin (TTX), one of the most toxic substances in nature, is present in bacteria, invertebrates, fishes, and amphibians. Marine organisms seem to bioaccumulate TTX from their food or acquire it from symbiotic bacteria, but its origin in amphibians is unclear. Taricha granulosa can exhibit high TTX levels, presumably concentrated in skin poison glands, acting as an agent of selection upon predatory garter snakes (Thamnophis). This co-evolutionary arms race induces variation in T. granulosa TTX levels, from very high to undetectable. Using morphology and biochemistry, we investigated differences in toxin localization and quality between two populations at the extremes of toxicity. TTX concentration within poison glands is related to the volume of a single cell type in which TTX occurs exclusively in distinctive secretory granules, suggesting a relationship between granule structure and chemical composition. TTX was detected in mucous glands in both populations, contradicting the general understanding that these glands do not secrete defensive chemicals and expanding currently held interpretations of amphibian skin gland functionality. Skin secretions of the two populations differed in low-mass molecules and proteins. Our results demonstrate that interpopulation variation in TTX levels is related to poison gland morphology.

ACS Style

Pedro Luiz Mailho-Fontana; Carlos Jared; Marta Maria Antoniazzi; Juliana Mozer Sciani; Daniel Carvalho Pimenta; Amber N. Stokes; Taran Grant; Edmund D. Brodie; Edmund D Brodie; Edmund D. Brodie Jr.. Variations in tetrodotoxin levels in populations of Taricha granulosa are expressed in the morphology of their cutaneous glands. Scientific Reports 2019, 9, 1 -8.

AMA Style

Pedro Luiz Mailho-Fontana, Carlos Jared, Marta Maria Antoniazzi, Juliana Mozer Sciani, Daniel Carvalho Pimenta, Amber N. Stokes, Taran Grant, Edmund D. Brodie, Edmund D Brodie, Edmund D. Brodie Jr.. Variations in tetrodotoxin levels in populations of Taricha granulosa are expressed in the morphology of their cutaneous glands. Scientific Reports. 2019; 9 (1):1-8.

Chicago/Turabian Style

Pedro Luiz Mailho-Fontana; Carlos Jared; Marta Maria Antoniazzi; Juliana Mozer Sciani; Daniel Carvalho Pimenta; Amber N. Stokes; Taran Grant; Edmund D. Brodie; Edmund D Brodie; Edmund D. Brodie Jr.. 2019. "Variations in tetrodotoxin levels in populations of Taricha granulosa are expressed in the morphology of their cutaneous glands." Scientific Reports 9, no. 1: 1-8.

Journal article
Published: 01 October 2019 in Fish & Shellfish Immunology
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Stingrays skin secretions are largely studied due to the human envenoming medical relevance of the sting puncture that evolves to inflammatory events, including necrosis. Such toxic effects can be correlated to the biochemical composition of the sting mucus, according to the literature. Fish skin plays important biological roles, such as the control of the osmotic pressure gradient, protection against mechanical forces and microorganism infections. The mucus, on the other hand, is a rich and complex fluid, acting on swimming, nutrition and the innate immune system. The elasmobranch's epidermis is a tissue composed mainly by mucus secretory cells, and marine stingrays have already been described to present secretory glands spread throughout the body. Little is known about the biochemical composition of the stingray mucus, but recent studies have corroborated the importance of mucus in the envenomation process. Aiming to assess the mucus composition, a new non-invasive mucus collection method was developed that focused on peptides and proteins, and biological assays were performed to analyze the toxic and immune activities of the Hypanus americanus mucus. Pathophysiological characterization showed the presence of peptidases on the mucus, as well as the induction of edema and leukocyte recruitment in mice. The fractionated mucus improved phagocytosis on macrophages and showed antimicrobial activity against T. rubrumç. neoformans and C. albicans in vitro. The proteomic analyses showed the presence of immune-related proteins like actin, histones, hemoglobin, and ribosomal proteins. This protein pattern is similar to those reported for other fish mucus and stingray venoms. This is the first report depicting the Hypanus stingray mucus composition, highlighting its biochemical composition and importance for the stingray immune system and the possible role on the envenomation process.

ACS Style

Guilherme Rabelo Coelho; Pedro Prezotto Neto; Fernanda Cortinhas Barbosa; Rafael Silva Dos Santos; Patrícia Brigatte; Patrick Jack Spencer; Sandra Coccuzzo Sampaio; Fernanda D’Amélio; Daniel Carvalho Pimenta; Juliana Mozer Sciani. Biochemical and biological characterization of the Hypanus americanus mucus: A perspective on stingray immunity and toxins. Fish & Shellfish Immunology 2019, 93, 832 -840.

AMA Style

Guilherme Rabelo Coelho, Pedro Prezotto Neto, Fernanda Cortinhas Barbosa, Rafael Silva Dos Santos, Patrícia Brigatte, Patrick Jack Spencer, Sandra Coccuzzo Sampaio, Fernanda D’Amélio, Daniel Carvalho Pimenta, Juliana Mozer Sciani. Biochemical and biological characterization of the Hypanus americanus mucus: A perspective on stingray immunity and toxins. Fish & Shellfish Immunology. 2019; 93 ():832-840.

Chicago/Turabian Style

Guilherme Rabelo Coelho; Pedro Prezotto Neto; Fernanda Cortinhas Barbosa; Rafael Silva Dos Santos; Patrícia Brigatte; Patrick Jack Spencer; Sandra Coccuzzo Sampaio; Fernanda D’Amélio; Daniel Carvalho Pimenta; Juliana Mozer Sciani. 2019. "Biochemical and biological characterization of the Hypanus americanus mucus: A perspective on stingray immunity and toxins." Fish & Shellfish Immunology 93, no. : 832-840.

Journal article
Published: 14 September 2019 in Journal of Proteomics
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Amphibians display a toxic secretion that works as chemical defenses against predators and/or microorganisms that is stored in specialized glands located in the tegument. For some animals, such glands have accumulated in specific regions of the body and formed prominent structures known as macroglands. The Bufonidae family displays conspicuous macroglands in a post-orbital position, termed parotoids, which secretions are known to be extremely viscous and rich in alkaloids and steroids. Few proteins have been described in this material, though. Mainly, because of the difficulties to handle such biological matrix. In this context, we have performed a proteomic study on the parotoid macrogland secretion of the Asian bufonid Duttaphrynus melanostictus. By employing the Ion-Exchange (IEx)-batch chromatography (anionic, cationic and both) we obtained six fractions - bound and unbound - that were submitted to an in solution-trypsin digestion followed by LC-MS/MS. Proteins related to: antioxidant activity, binding processes (carbohydrate/lipid/protein), energy metabolism, hydrolases, lipid metabolism and membrane traffic were identified. Moreover, IEx was able to preserve the biological activity of the retrieved proteins (peptidasic). The current study increases the knowledge on the proteins present in the bufonids parotoid macrogland secretion, providing a better understanding of the physiological role played by such molecules. SIGNIFICANCE: The current approach allowed a detailed proteomic analysis of the several proteins synthesized in the D. melanostictus parotoid macrogland (Bufonidae) that are secreted into the skins, but embedded within a complex viscous biological matrix. Moreover, our results aim to increase the knowledge on the biological role played by such proteins at the skin.

ACS Style

Douglas O.C. Mariano; José Pedro Prezotto-Neto; Patrick J. Spencer; Juliana Sciani; Daniel C. Pimenta. Proteomic analysis of soluble proteins retrieved from Duttaphrynus melanostictus skin secretion by IEx-batch sample preparation. Journal of Proteomics 2019, 209, 103525 .

AMA Style

Douglas O.C. Mariano, José Pedro Prezotto-Neto, Patrick J. Spencer, Juliana Sciani, Daniel C. Pimenta. Proteomic analysis of soluble proteins retrieved from Duttaphrynus melanostictus skin secretion by IEx-batch sample preparation. Journal of Proteomics. 2019; 209 ():103525.

Chicago/Turabian Style

Douglas O.C. Mariano; José Pedro Prezotto-Neto; Patrick J. Spencer; Juliana Sciani; Daniel C. Pimenta. 2019. "Proteomic analysis of soluble proteins retrieved from Duttaphrynus melanostictus skin secretion by IEx-batch sample preparation." Journal of Proteomics 209, no. : 103525.

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.

Corrigendum
Published: 27 May 2019 in BioMed Research International
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[This corrects the article DOI: 10.1155/2019/2492315.].

ACS Style

Daniel S. S. De Medeiros; Tiago B. Rego; Ana P. De A. Dos Santos; Adriana S. Pontes; Leandro S. Moreira-Dill; Najla B. Matos; Juliana P. Zuliani; Rodrigo G. Stábeli; Carolina B. G. Teles; Andreimar Soares; Angelo R. De M. Sperotto; Dinara J. Moura; Jenifer Saffi; Cleópatra Alves Da Silva Caldeira; Daniel Carvalho Pimenta; Leonardo A. Calderon. Corrigendum to “Biochemical and Biological Profile of Parotoid Secretion of the Amazonian Rhinella marina (Anura: Bufonidae)”. BioMed Research International 2019, 2019, 1 -1.

AMA Style

Daniel S. S. De Medeiros, Tiago B. Rego, Ana P. De A. Dos Santos, Adriana S. Pontes, Leandro S. Moreira-Dill, Najla B. Matos, Juliana P. Zuliani, Rodrigo G. Stábeli, Carolina B. G. Teles, Andreimar Soares, Angelo R. De M. Sperotto, Dinara J. Moura, Jenifer Saffi, Cleópatra Alves Da Silva Caldeira, Daniel Carvalho Pimenta, Leonardo A. Calderon. Corrigendum to “Biochemical and Biological Profile of Parotoid Secretion of the Amazonian Rhinella marina (Anura: Bufonidae)”. BioMed Research International. 2019; 2019 ():1-1.

Chicago/Turabian Style

Daniel S. S. De Medeiros; Tiago B. Rego; Ana P. De A. Dos Santos; Adriana S. Pontes; Leandro S. Moreira-Dill; Najla B. Matos; Juliana P. Zuliani; Rodrigo G. Stábeli; Carolina B. G. Teles; Andreimar Soares; Angelo R. De M. Sperotto; Dinara J. Moura; Jenifer Saffi; Cleópatra Alves Da Silva Caldeira; Daniel Carvalho Pimenta; Leonardo A. Calderon. 2019. "Corrigendum to “Biochemical and Biological Profile of Parotoid Secretion of the Amazonian Rhinella marina (Anura: Bufonidae)”." BioMed Research International 2019, no. : 1-1.

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.

Research article
Published: 21 February 2019 in BioMed Research International
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Skin secretions of frogs have a high chemical complexity. They have diverse types of biomolecules, such as proteins, peptides, biogenic amines, and alkaloids. These compounds protect amphibians’ skin against growth of bacteria, fungi, and protozoa and participate in defense system against attack from predators. Therewith, this work performed biochemical and biological profile of macroglands parotoid secretion from cane toad. For poison analysis, we performed molecular exclusion and reverse phase chromatography, electrophoresis, and mass spectrometry. Antimicrobial, antiplasmodial, leishmanicidal, cytotoxicity, genotoxicity, and inflammatory activity of crude and/or fractions ofR. marinasecretion were also evaluated. Fractionation prior to filtration from poison showed separation of low mass content (steroids and alkaloids) and high molecular mass (protein). Material below 10 kDa two steroids, marinobufagin and desacetylcinobufagin, was detected. Crude extract and fractions were active againstStaphylococcus aureus,Pseudomonas aeruginosa,Escherichia coli,Plasmodium falciparum,Leishmania guyanensis, andLeishmania braziliensis. Crude extract was also active against cancer cells although it was not cytotoxic for normal cells. This extract did not show significant DNA damage but it showed an important inflammatory effect in vivo. The information obtained in this work contributes to the understanding of the constituents ofR.marinasecretion as well as the bioactive potential of these molecules.

ACS Style

Daniel S. S. de Medeiros; Tiago B. Rego; Ana P. De A. Dos Santos; Adriana S. Pontes; Leandro S. Moreira-Dill; Najla B. Matos; Juliana P. Zuliani; Rodrigo G. Stábeli; Carolina B. G. Teles; Andreimar Soares; Angelo R. De M. Sperotto; Dinara J. Moura; Jenifer Saffi; Cleópatra Alves Da Silva Caldeira; Daniel Carvalho Pimenta; Leonardo A. Calderon. Biochemical and Biological Profile of Parotoid Secretion of the AmazonianRhinella marina(Anura: Bufonidae). BioMed Research International 2019, 2019, 1 -15.

AMA Style

Daniel S. S. de Medeiros, Tiago B. Rego, Ana P. De A. Dos Santos, Adriana S. Pontes, Leandro S. Moreira-Dill, Najla B. Matos, Juliana P. Zuliani, Rodrigo G. Stábeli, Carolina B. G. Teles, Andreimar Soares, Angelo R. De M. Sperotto, Dinara J. Moura, Jenifer Saffi, Cleópatra Alves Da Silva Caldeira, Daniel Carvalho Pimenta, Leonardo A. Calderon. Biochemical and Biological Profile of Parotoid Secretion of the AmazonianRhinella marina(Anura: Bufonidae). BioMed Research International. 2019; 2019 ():1-15.

Chicago/Turabian Style

Daniel S. S. de Medeiros; Tiago B. Rego; Ana P. De A. Dos Santos; Adriana S. Pontes; Leandro S. Moreira-Dill; Najla B. Matos; Juliana P. Zuliani; Rodrigo G. Stábeli; Carolina B. G. Teles; Andreimar Soares; Angelo R. De M. Sperotto; Dinara J. Moura; Jenifer Saffi; Cleópatra Alves Da Silva Caldeira; Daniel Carvalho Pimenta; Leonardo A. Calderon. 2019. "Biochemical and Biological Profile of Parotoid Secretion of the AmazonianRhinella marina(Anura: Bufonidae)." BioMed Research International 2019, no. : 1-15.

Article
Published: 29 January 2019 in The Protein Journal
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Amphibians are, currently, considered the first vertebrates that had performed the aquatic to terrestrial transition during evolution; therefore, water balance and dehydration control were prerequisites for such environment conquering. Among anurans, Phyllomedusa is a well-studied genus, due to its peptide-rich skin secretion. Here, we have analyzed the skin secretion of Phyllomedusa distincta targeting the proteins present in the skin secretion. The major soluble protein was chromatographically isolated and utilized to immunize rabbits. Through proteomics approaches, we were able to identify such protein as being the diacylglycerol O-acyltransferase 2 (DGAT2), a crucial enzyme involved in lipid synthesis and in the skin water balance. Immunohistochemistry assays revealed the protein tissular distribution for different animal species, belonging to different branches of the phylogenetic tree. Specifically, there was positivity to the anti-DGAT2 on Amphibians’ skin, and no antibody recognition on fish and mammals’ skins. The DGAT2 multiple sequence alignment reveals some degree of conservation throughout the genera; however, there is a different cysteine pattern among them. Molecular modeling analyses corroborate that the different cysteine pattern leads to distinct 3D structures, explaining the different antibody recognition. Moreover, the protein phylogenetic analyses place the Xenopus DGAT2 (the available amphibian representative) next to the Coelacanthus enzyme, which have led the authors to term this a ‘paleo-protein’. DGAT2 would be, therefore, an ancient protein, crucial to the terrestrial environment conquest, with a unique folding—as indicated by the molecular models and immunohistochemistry analyses—a consequence of the different cysteine pattern but with conserved biological function.

ACS Style

Juliana M. Sciani; Adriana Neves; Ruth C. Vassão; Patrick Spencer; Marta M. Antoniazzi; Carlos Jared; Daniel C. Pimenta. The Amphibian Diacylglycerol O-acyltransferase 2 (DGAT2): a ‘paleo-protein’ with Conserved Function but Unique Folding. The Protein Journal 2019, 38, 83 -94.

AMA Style

Juliana M. Sciani, Adriana Neves, Ruth C. Vassão, Patrick Spencer, Marta M. Antoniazzi, Carlos Jared, Daniel C. Pimenta. The Amphibian Diacylglycerol O-acyltransferase 2 (DGAT2): a ‘paleo-protein’ with Conserved Function but Unique Folding. The Protein Journal. 2019; 38 (1):83-94.

Chicago/Turabian Style

Juliana M. Sciani; Adriana Neves; Ruth C. Vassão; Patrick Spencer; Marta M. Antoniazzi; Carlos Jared; Daniel C. Pimenta. 2019. "The Amphibian Diacylglycerol O-acyltransferase 2 (DGAT2): a ‘paleo-protein’ with Conserved Function but Unique Folding." The Protein Journal 38, no. 1: 83-94.

Journal article
Published: 02 January 2019 in New Journal of Chemistry
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The synthesis of amidoglucals and glucal esters in good to high yields using 2-iodo-3,4,6-tri-O-acetyl-d-glucal as a substrate, Mo(CO)6 as a carbon monoxide source and PdCl2 as a catalyst is reported.

ACS Style

Mariana P. Darbem; Karina S. Kanno; Isadora Maria de Oliveira; C. Henrique A. Esteves; Daniel C. Pimenta; Hélio A. Stefani. Synthesis of amidoglucals and glucal esters via carbonylative coupling reactions of 2-iodoglucal using Mo(CO)6 as a CO source. New Journal of Chemistry 2019, 43, 696 -699.

AMA Style

Mariana P. Darbem, Karina S. Kanno, Isadora Maria de Oliveira, C. Henrique A. Esteves, Daniel C. Pimenta, Hélio A. Stefani. Synthesis of amidoglucals and glucal esters via carbonylative coupling reactions of 2-iodoglucal using Mo(CO)6 as a CO source. New Journal of Chemistry. 2019; 43 (2):696-699.

Chicago/Turabian Style

Mariana P. Darbem; Karina S. Kanno; Isadora Maria de Oliveira; C. Henrique A. Esteves; Daniel C. Pimenta; Hélio A. Stefani. 2019. "Synthesis of amidoglucals and glucal esters via carbonylative coupling reactions of 2-iodoglucal using Mo(CO)6 as a CO source." New Journal of Chemistry 43, no. 2: 696-699.