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Mygalin, a diacylspermidine that is naturally found in the hemolymph of the spider Acanthoscurria gomesiana, is of interest for development as a potential analgesic. Previous studies have shown that acylpolyamines modulate glutamatergic receptors with the potential to alter pain pathways. This study aimed to evaluate the effects of mygalin on acute and chronic pain in rodents. For evaluation of acute pain, Wistar rats were subjected to tail-flick and hot-plate nociceptive tests. For the evaluation of chronic neuropathic pain, a partial ligation of the sciatic nerve was performed and, 21 days later, animals were examined in hot-plate, tail-flick, acetone, and von Frey tests. Either Mygalin or vehicle was microinjected in the dorsal raphe nucleus (DRN) before the tests. Another group was pretreated with selective antagonists of glutamate receptors (LY 235959, MK-801, CNQX, and NBQX). Mygalin decreases nociceptive thresholds on both acute and chronic neuropathic pain models in all the tests performed. The lowest dose of mygalin yielded the most effective nociception, showing an increase of 63% of the nociceptive threshold of animals with neuropathic chronic pain. In conclusion, mygalin microinjection in the DRN results in antinociceptive effect in models of neuropathic pain, suggesting that acylpolyamines and their derivatives, such as this diacylspermidine, could be pursued for the treatment of neuropathic pain and development of selective analgesics.
Ana C. Medeiros; Priscila Medeiros; Renato L. de Freitas; Pedro Ismael Da Silva Júnior; Norberto C. Coimbra; Wagner F. dos Santos. Acanthoscurria gomesiana spider‐derived synthetic mygalin in the dorsal raphe nucleus modulates acute and chronic pain. Journal of Biochemical and Molecular Toxicology 2021, e22877 .
AMA StyleAna C. Medeiros, Priscila Medeiros, Renato L. de Freitas, Pedro Ismael Da Silva Júnior, Norberto C. Coimbra, Wagner F. dos Santos. Acanthoscurria gomesiana spider‐derived synthetic mygalin in the dorsal raphe nucleus modulates acute and chronic pain. Journal of Biochemical and Molecular Toxicology. 2021; ():e22877.
Chicago/Turabian StyleAna C. Medeiros; Priscila Medeiros; Renato L. de Freitas; Pedro Ismael Da Silva Júnior; Norberto C. Coimbra; Wagner F. dos Santos. 2021. "Acanthoscurria gomesiana spider‐derived synthetic mygalin in the dorsal raphe nucleus modulates acute and chronic pain." Journal of Biochemical and Molecular Toxicology , no. : e22877.
Infectious diseases are among the major causes of death in the human population. A wide variety of organisms produce antimicrobial peptides (AMPs) as part of their first line of defense. A peptide from Acanthoscurria rondoniae plasma, rondonin – with antifungal activity, a molecular mass of 1,236 Da and primary sequence IIIQYEGHKH – was previously studied (Uniprot accession number B3EWP8). It showed identity with the C-terminus of subunit "D" of the hemocyanin of the Aphonopelma hentzi spider. This result led us to propose a new pathway of the immune system of arachnids that suggests a new function to hemocyanin: production of antimicrobial peptides. Rondonin does not interact with model membranes and was able to bind to yeast nucleic acids but not bacteria. It was not cytotoxic against mammalian cells. The antifungal activity of rondonin is pH dependent and peaks at pH ~4-5. The peptide presents synergism with Gomesin (spider hemocyte antimicrobial peptide - UniProtKB - P82358) against human yeast pathogens, suggesting a new potential alternative treatment option. Antiviral activity was detected against RNA viruses, measles, H1N1 and encephalomyocarditis. This is the first report of an arthropod hemocyanin fragment with activity against human viruses. Currently, it is vital to invest in the search for natural and synthetic antimicrobial compounds that, above all, present alternative mechanisms of action to first-choice antimicrobials.
Katie C. T. Riciluca; Ursula C. Oliveira; Ronaldo Z. Mendonça; José C. Bozelli Junior; Shirley Schreier; Pedro I Da Silva Junior. Rondonin: antimicrobial properties and mechanism of action. FEBS Open Bio 2021, 1 .
AMA StyleKatie C. T. Riciluca, Ursula C. Oliveira, Ronaldo Z. Mendonça, José C. Bozelli Junior, Shirley Schreier, Pedro I Da Silva Junior. Rondonin: antimicrobial properties and mechanism of action. FEBS Open Bio. 2021; ():1.
Chicago/Turabian StyleKatie C. T. Riciluca; Ursula C. Oliveira; Ronaldo Z. Mendonça; José C. Bozelli Junior; Shirley Schreier; Pedro I Da Silva Junior. 2021. "Rondonin: antimicrobial properties and mechanism of action." FEBS Open Bio , no. : 1.
Antimicrobial peptides have been a major research subject since the rise of antimicrobial resistance as a major public health problem. These molecules are considered a potential therapeutic source of antibiotics with broad-spectrum activity against microorganisms. Two antimicrobial peptides were isolated from the mucus of the Limacus flavus slug. The mucus was obtained by thermal shock, lyophilized and extracted with acetic acid. The supernatant was prefractionated in Sep-Pak and shortly thereafter fractionated by reverse-phase high-performance liquid chromatography. The manually obtained fractions were used in antimicrobial and cytotoxic assays and finally subjected to mass spectrometry (MS/MS). Characterization was performed by bioinformatics analysis with the tool Peaks®X + and by comparison with the NCBI and UniProt-SwissProt databases. Additionally, the physicochemical parameters of the samples were evaluated with online programs. Two fractions comtained antimicrobial peptides with the ability to inhibit Micrococcus luteus A270; both samples, LFMP-001 and LFMP-002, were hydrophilic molecules consisting of fewer than 20 residues. Comparison of the SDS-PAGE and Peaks®X + data showed that both had Mw < 3 kDa. In summary, this study presents data on the isolation and characterization of antimicrobial peptides from a slug and shows their potential against gram-positive bacteria.
Patricia Yumi Hayashida; Pedro Ismael Da Silva Junior. Insights into Antimicrobial Peptides from Limacus flavus Mucus. Current Microbiology 2021, 1 -10.
AMA StylePatricia Yumi Hayashida, Pedro Ismael Da Silva Junior. Insights into Antimicrobial Peptides from Limacus flavus Mucus. Current Microbiology. 2021; ():1-10.
Chicago/Turabian StylePatricia Yumi Hayashida; Pedro Ismael Da Silva Junior. 2021. "Insights into Antimicrobial Peptides from Limacus flavus Mucus." Current Microbiology , no. : 1-10.
Background Snails and slugs were used as a treatment for many health problems therefore ancient times. Since the antimicrobial resistance became a major global thread, antimicrobial peptides have been considered as a potential source for development of new drugs, especially for drug-resistant bacteria. Nowadays reports confirm that the mucous secretions have antimicrobial, antiviral and antifungal properties. Methods The present study has the objective to characterize and evaluate antimicrobial peptides of Limacus flavus mucus. The mucus was obtained by thermal shock and submitted to RP-HPLC. Fractions were used to perform the antimicrobial activity and hemolytic assays, electrophoresis (SDS-Page Gel) and submitted to mass spectrometry (LC-MS / MS). Identification and characterization was performed by PeaksX+ software. The physicochemical parameters were evaluated with bioinformatics tools, which predicted water solubility, iso-electric point, charge net and its primary structure. Results Three fractions were isolated from the mucus of L. flavus and presented antifungal and antibacterial activity. The mucus showed greater inhibition for filamentous fungi (Aspergillus niger), yeast (Cryptococcus neoformans), Gram positive bacteria (Bacillus subtilis, Micrococcus luteus) and Gram negative bacteria (Enterobacter cloacae). These fractions also did not show hemolytic activity for human blood cells (erythrocytes). Fraction’s sequences were identified and presents Mw <3kDa, WLGH, DLQW, YLRW, respectively. Conclusion This study revealed three antimicrobial peptides of L. flavus mucus with a wide range of antimicrobial activity and its physic-chemical characterization.
Patricia Yumi Hayashida; Pedro Ismael Da Silva Júnior. Limacus flavus yellow slug: bioactive molecules in the mucus. 2021, 1 .
AMA StylePatricia Yumi Hayashida, Pedro Ismael Da Silva Júnior. Limacus flavus yellow slug: bioactive molecules in the mucus. . 2021; ():1.
Chicago/Turabian StylePatricia Yumi Hayashida; Pedro Ismael Da Silva Júnior. 2021. "Limacus flavus yellow slug: bioactive molecules in the mucus." , no. : 1.
Millipedes are among the most diverse and abundant arthropods in terrestrial environments. However, little is known about their innate immune response against invading pathogenic microorganisms, which is very intriguing considering that the evolutionary success of millipedes is largely due to this complex and primitive defense system, since it allowed them to colonize a wide variety of microhabitats characterized by their high microbial proliferation. Accordingly, the aim of the present work was to determine the presence of antimicrobial peptides in the hemolymph of the millipede Rhinocricus sp. In total, four native peptides with potent antimicrobial activity against different microorganisms, lack of cytotoxicity against Vero cells and lack of hemolytic effects against human erythrocytes were isolated and named RP40–16, RP40–19, RP40–20/1 and RP40–20/2. The analysis with bioinformatics tools suggested that these peptides may be encrypted in large proteins present in the plasma: Hemocyanin and thioester-containing protein. Considering these results, it can be said that millipede hemolymph represents a promising source of molecules with potential for the development of non-conventional antibiotics. Therefore, in order to have a clearer notion of the biotechnological potential and the role of these peptides in the innate immune response of Rhinocricus sp., future studies should focus on elucidating their mechanisms of action, as well as additional biological properties
Paula J. Segura-Ramírez; Pedro Machado de Godoy; Ivan N. Avino; Pedro I. Silva Junior. Encrypted antimicrobial peptides from proteins present in the plasma of the millipede Rhinocricus sp. Journal of Proteomics 2021, 242, 104239 .
AMA StylePaula J. Segura-Ramírez, Pedro Machado de Godoy, Ivan N. Avino, Pedro I. Silva Junior. Encrypted antimicrobial peptides from proteins present in the plasma of the millipede Rhinocricus sp. Journal of Proteomics. 2021; 242 ():104239.
Chicago/Turabian StylePaula J. Segura-Ramírez; Pedro Machado de Godoy; Ivan N. Avino; Pedro I. Silva Junior. 2021. "Encrypted antimicrobial peptides from proteins present in the plasma of the millipede Rhinocricus sp." Journal of Proteomics 242, no. : 104239.
Background: Tarantulas are the most common invertebrates pets, especially in North America and Europe. The most commercialized genera are from Southern Asia and tropical Americas, represented by Vitalius, found in Southeastern Brazil, and Brachypelma, common in Mexican desert. Bites by these spiders in humans occurs during manipulation and generally result in clinical manifestations such local pain, erythema and oedema, with the possibility of secondary local infection. Hence, the cases are usually treated with prescription free drugs such as antihistamines and anti-inflammatories. Methods: In this work, we investigated the post treatment with commercial nonsteroidal and steroidal anti-inflammatories and anti-histamines administered by oral and intraperitoneal routes on rat paw oedema induced by venoms of V. dubius and B. smithi. Hydroplethysmometer standard oedema measurement and Evans blue extravasation were performed. Dose standardization experiments showed that the V. dubius is more potent than B. smithi, and doses were established at 30 μg/paw and 60 μg/paw respectively. Results: The oral post-administration of ketoprofen (non-selective cyclooxygenase inhibitor) and prednisolone (steroidal anti-inflammatory) markedly reduced a paw oedema evoked by only V. dubius venom, but loratadine (H1-antihistamine) had negligible effect on rat paw oedema induced by both venoms. Intraperitoneal administration, ketoprofen (20 mg Kg-1) and loratadine (5 mg Kg-1) reduced the rat paw oedema induced by V. dubius and B. smithi while methylprednisolone (10 mg Kg-1) only inhibited the oedema induced by V. dubius. Conclusion: These results suggest that the pos-treatment with nonsteroidal and steroidal anti-inflammatory drugs are more potent than antihistamines in attenuating the local effect induced by V. dubius and B. smithi venoms.
Bruno Ricardo Alves; Rafael Sutti; Pedro Ismael da Silva; Rogerio Bertani; Jair Guilherme Santos-Junior; Thomaz Augusto Alves Rocha e Silva; Alessandra Linardi. Non-clinical evidence supports anti-inflammatories as more effective medication than antihistamines against tarantula local effects envenomation. 2021, 1 .
AMA StyleBruno Ricardo Alves, Rafael Sutti, Pedro Ismael da Silva, Rogerio Bertani, Jair Guilherme Santos-Junior, Thomaz Augusto Alves Rocha e Silva, Alessandra Linardi. Non-clinical evidence supports anti-inflammatories as more effective medication than antihistamines against tarantula local effects envenomation. . 2021; ():1.
Chicago/Turabian StyleBruno Ricardo Alves; Rafael Sutti; Pedro Ismael da Silva; Rogerio Bertani; Jair Guilherme Santos-Junior; Thomaz Augusto Alves Rocha e Silva; Alessandra Linardi. 2021. "Non-clinical evidence supports anti-inflammatories as more effective medication than antihistamines against tarantula local effects envenomation." , no. : 1.
Spiders are the most successful and diversified group of venomous animals. Currently, there are more than 49,000 species distributed almost all over the world. This broad distribution suggests that they have efficient strategies to improve their survival; one of them is the production of highly elaborate venoms, which are a heterogeneous mixture of molecules like inorganic salts, peptides, proteins, and enzymes. Considering this, this study aimed to analyze the venom of the spider Avicularia juruensis (Mygalomorphae: Theraphosidae) searching for proteolytic enzymes. Using zymography, electrophoresis, transcriptomics and proteomics approaches we identified one neprilysin able to degrade casein, that we named “Ajur_Neprilysin”. Neprilysins are metalloendopeptidases whose presence has already been described in animal venoms, however, its function has not yet been elucidated. Our results showed for the first time one non-bacterial neprilysin which can cleave casein and suggest that its role in envenomation is to degrade the extracellular matrix, facilitating the access of other toxins to their targets, as well as digestive fluids. Moreover, this discovery contributes to increasing the knowledge about little-studied species, since the Ajur_Neprilysin is the second neprilysin found in the venom from a mygalomorph spider.
Soraia Maria Do Nascimento; Ursula Castro De Oliveira; Milton Yutaka Nishiyama-Jr; Alexandre Keiji Tashima; Pedro Ismael Da Silva Junior. Presence of a neprilysin on Avicularia juruensis (Mygalomorphae: Theraphosidae) venom. Toxin Reviews 2021, 1 -10.
AMA StyleSoraia Maria Do Nascimento, Ursula Castro De Oliveira, Milton Yutaka Nishiyama-Jr, Alexandre Keiji Tashima, Pedro Ismael Da Silva Junior. Presence of a neprilysin on Avicularia juruensis (Mygalomorphae: Theraphosidae) venom. Toxin Reviews. 2021; ():1-10.
Chicago/Turabian StyleSoraia Maria Do Nascimento; Ursula Castro De Oliveira; Milton Yutaka Nishiyama-Jr; Alexandre Keiji Tashima; Pedro Ismael Da Silva Junior. 2021. "Presence of a neprilysin on Avicularia juruensis (Mygalomorphae: Theraphosidae) venom." Toxin Reviews , no. : 1-10.
Toll-like receptors (TLRs) are transmembrane proteins that are key regulators of innate and adaptive immune responses, particularly TLR4, and they have been identified as potential drug targets for the treatment of disease. Several low-molecular-weight compounds are being considered as new drug targets for various applications, including as immune modulators. Mygalin, a 417 Da synthetic bis-acylpolyamine, is an analog of spermidine that has microbicidal activity. In this study, we investigated the effect of mygalin on the innate immune response based on a virtual screening (VS) and molecular docking analysis. Bone marrow-derived macrophages and the cell lines J774A.1 and RAW 264.7 stimulated with lipopolysaccharide (LPS) were used to confirm the data obtained in silico. Virtual screening and molecular docking suggested that mygalin binds to TLR4 via the protein myeloid differentiation factor 2 (MD-2) and LPS. Macrophages stimulated by mygalin plus LPS showed suppressed gene expression of tumor necrosis factor (TNF-α), interleukine 6 (IL-6), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), as well as inhibition of signaling protein p65 of the nuclear factor κB (NF-κB), resulting in decreased production of nitric oxide (NO) and TNF-α. These results indicate that mygalin has anti-inflammatory potential, being an attractive option to be explored. In addition, we reinforce the importance of virtual screening analysis to assist in the discovery of new drugs.
Abraham Espinoza-Culupú; Ricardo Vázquez-Ramírez; Mariella Farfán-López; Elizabeth Mendes; Maria Notomi Sato; Pedro Ismael Da Silva Junior; Monamaris Marques Borges. Acylpolyamine Mygalin as a TLR4 Antagonist Based on Molecular Docking and In Vitro Analyses. Biomolecules 2020, 10, 1624 .
AMA StyleAbraham Espinoza-Culupú, Ricardo Vázquez-Ramírez, Mariella Farfán-López, Elizabeth Mendes, Maria Notomi Sato, Pedro Ismael Da Silva Junior, Monamaris Marques Borges. Acylpolyamine Mygalin as a TLR4 Antagonist Based on Molecular Docking and In Vitro Analyses. Biomolecules. 2020; 10 (12):1624.
Chicago/Turabian StyleAbraham Espinoza-Culupú; Ricardo Vázquez-Ramírez; Mariella Farfán-López; Elizabeth Mendes; Maria Notomi Sato; Pedro Ismael Da Silva Junior; Monamaris Marques Borges. 2020. "Acylpolyamine Mygalin as a TLR4 Antagonist Based on Molecular Docking and In Vitro Analyses." Biomolecules 10, no. 12: 1624.
Sea urchins live in a challenging environment that requires rapid and efficient responses against pathogens and invaders. This response may be also important in reproductive processes once males and females release their gametes into water. In addition, the gonads are organs with dual function: reproductive organ and nutrient reserve, therefore it needs efficient protective mechanisms to preserve the nutrients as well as the reproductive cells. The aim of this study was to evaluate the presence and characterize antimicrobial molecules in the male and female gonads of the sea urchin Lytechinus variegatus. Through HPLC purification, antimicrobial activity test and mass spectrometry several antimicrobial molecules were found in the gonads of both gender. Computational in silico analyses showed that they are fragments of a glycoprotein called toposome, also known as major yolk protein (MYP) which is one of the major proteins found in the gonads. Although different functions have been reported for this protein, this is the first description of a direct antimicrobial activity in Lytechinus variegatus. The results indicate that when undergoing proteolysis the toposome generates different fragments with antimicrobial activity which may indicate the importance of a rapid defense response strategy against invading microorganisms in the gonads used by both males and females sea urchins.
Dal Figueiredo; Ws Santos; Fabio Montoni; Leo Kei Iwai; Pi Silva Junior. Toposome: Source of antimicrobial molecules in the gonads of the sea urchin Lytechinus variegatus (Lamarck, 1816). Fish & Shellfish Immunology 2020, 109, 51 -61.
AMA StyleDal Figueiredo, Ws Santos, Fabio Montoni, Leo Kei Iwai, Pi Silva Junior. Toposome: Source of antimicrobial molecules in the gonads of the sea urchin Lytechinus variegatus (Lamarck, 1816). Fish & Shellfish Immunology. 2020; 109 ():51-61.
Chicago/Turabian StyleDal Figueiredo; Ws Santos; Fabio Montoni; Leo Kei Iwai; Pi Silva Junior. 2020. "Toposome: Source of antimicrobial molecules in the gonads of the sea urchin Lytechinus variegatus (Lamarck, 1816)." Fish & Shellfish Immunology 109, no. : 51-61.
Hemoglobin is one of the most important molecules of the human body. Beyond its physiological activity, hemoglobins are able to inhibit the growth of several microorganisms. Since 1999, studies have reported that antimicrobial peptides can be produced by blood-feeding insects through hemoglobin digestion, and it has been reported that Triatoma infestans can generate an antimicrobial fragment from human fibrinopeptide. Thus T. infestans intestinal content was analyzed through Reverse Phase High-Performance Liquid Chromatography (RP-HPLC), the eluted fractions were tested against Micrococcus luteus, Escherichia coli and Staphylococcus aureus, and the active fractions submitted to mass spectrometry. The data obtained were compared to hemoglobin databases to verify the presence of hemoglobin-derived fragments. Ten fractions eluted from chromatography presented antimicrobial activity, and when analyzed through mass spectrometry revealed the presence of 8 murine hemoglobin α-chain fragments and 24 fragments from murine hemoglobin β fragments. Through the compilation of the fragments is possible to obtain over 67% coverage of both sequences. Part of the amino acid sequences corresponds to the sequences already identified on other intestinal contents of arthropods, and are highly conserved between the blood of other wild animals that are the most common intermediate hosts of Chagas’ disease in Brazil and some of the main natural blood source for triatomines.
Laura Cristina Lima Diniz; Pedro Ismael Da Silva Junior. Hemoglobin Reassembly of Antimicrobial Fragments from the Midgut of Triatoma infestans. Biomolecules 2020, 10, 261 .
AMA StyleLaura Cristina Lima Diniz, Pedro Ismael Da Silva Junior. Hemoglobin Reassembly of Antimicrobial Fragments from the Midgut of Triatoma infestans. Biomolecules. 2020; 10 (2):261.
Chicago/Turabian StyleLaura Cristina Lima Diniz; Pedro Ismael Da Silva Junior. 2020. "Hemoglobin Reassembly of Antimicrobial Fragments from the Midgut of Triatoma infestans." Biomolecules 10, no. 2: 261.
Antimicrobial peptides and proteins (AMPs) are molecules that can interact with microbial cells and lead to membrane disruption or intracellular molecule interactions and death. Several molecules with antimicrobial effects also present other biological activities. One such protein group representing the duplicity of activities is the tachykinin family. Tachykinins (TKs) form a family of neuropeptides in vertebrates with a consensus C-terminal region (F-X-G-Y-R-NH2). Invertebrate TKs and TK-related peptides (TKRPs) are subfamilies found in invertebrates that present high homology with TKs and have similar biological effects. Several of these molecules have already been described but reports of TKRP in Hemiptera species are limited. By analyzing the Triatoma infestans hemolymph by reversed-phase high-performance liquid chromatography, biological assays, and mass spectrometry, two antimicrobial molecules were isolated and identified as TKRPs, which we named as TRP1-TINF and TRP2-TINF (tachykinin-related peptides I and II from T. infestans). TRP1-TINF is a random secondary structure peptide with 9 amino acid residues. It is susceptible to aminopeptidases degradation and is active mainly against Micrococcus luteus (32 μM). TRP2-TINF is a 10-amino acid peptide with a 310 helix secondary structure and is susceptible to carboxypeptidases degradation. It has major antimicrobial activity against both Pseudomonas aeruginosa and Escherichia coli (45 μM). Neither molecule is toxic to human erythrocytes and both present minor toxicity toward Vero cells at a concentration of 1000 μM. As the first description of TKRPs with antimicrobial activity in T. infestans, this work contributes to the wider comprehension of the insects’ physiology and describes pharmacological relevant molecules.
Laura Cristina Lima Diniz; Flávio Lopes Alves; Antonio Miranda; Pedro Ismael Da Silva Junior. Two Tachykinin-Related Peptides with Antimicrobial Activity Isolated from Triatoma infestans Hemolymph. Microbiology Insights 2020, 13, 1 .
AMA StyleLaura Cristina Lima Diniz, Flávio Lopes Alves, Antonio Miranda, Pedro Ismael Da Silva Junior. Two Tachykinin-Related Peptides with Antimicrobial Activity Isolated from Triatoma infestans Hemolymph. Microbiology Insights. 2020; 13 ():1.
Chicago/Turabian StyleLaura Cristina Lima Diniz; Flávio Lopes Alves; Antonio Miranda; Pedro Ismael Da Silva Junior. 2020. "Two Tachykinin-Related Peptides with Antimicrobial Activity Isolated from Triatoma infestans Hemolymph." Microbiology Insights 13, no. : 1.
Antimicrobial peptides (AMPs) are small molecules, which have a potential use as antibiotic or pharmacological tools. In chelicerate organisms, such as scorpions, these molecules constitute an alternative defense system against microorganisms. The aim of this work was to identify AMPs in the hemolymph of the Tityus serrulatus scorpion. Fractions of plasma and hemocytes were subjected to high-performance liquid chromatography (HPLC) and then analyzed to determine their activity in inhibiting microbial growth. One of the fractions from the hemocytes presents antimicrobial activity against microorganisms, such as Gram-negative and Gram-positive bacteria, fungi, and yeast. These fractions were analyzed by mass spectrometry, and a fragment of 3564 Da. was identified. The peptide was called serrulin, because it is derived from the species T. serrulatus. A comparison of the amino acid sequence of serrulin with databases shows that it has a similarity to the glycine-rich peptides described in Cupienius salai and Acanthoscurria gomesiana (spiders). Furthermore, serrulin has no hemolytic activity against human erythrocytes. While the presence of AMPs in T. serrulatus venom has been described in other works, this is the first work to characterize the presence of these molecules in the hemolymph (hemocytes) of this species and show its potential use as an alternative to conventional antibiotics against different species of microorganisms.
Thiago De Jesus Oliveira; Ursula Castro De Oliveira; Pedro Ismael Da Silva Junior. Serrulin: A Glycine-Rich Bioactive Peptide from the Hemolymph of the Yellow Tityus serrulatus Scorpion. Toxins 2019, 11, 517 .
AMA StyleThiago De Jesus Oliveira, Ursula Castro De Oliveira, Pedro Ismael Da Silva Junior. Serrulin: A Glycine-Rich Bioactive Peptide from the Hemolymph of the Yellow Tityus serrulatus Scorpion. Toxins. 2019; 11 (9):517.
Chicago/Turabian StyleThiago De Jesus Oliveira; Ursula Castro De Oliveira; Pedro Ismael Da Silva Junior. 2019. "Serrulin: A Glycine-Rich Bioactive Peptide from the Hemolymph of the Yellow Tityus serrulatus Scorpion." Toxins 11, no. 9: 517.
Antibiotic resistance is at dangerous levels and increasing worldwide. The search for new antimicrobial drugs to counteract this problem is a priority for health institutions and organizations, both globally and in individual countries. Sarconesiopsis magellanica blowfly larval excretions and secretions (ES) are an important source for isolating antimicrobial peptides (AMPs). This study aims to identify and characterize a new S. magellanica AMP. RP-HPLC was used to fractionate ES, using C18 columns, and their antimicrobial activity was evaluated. The peptide sequence of the fraction collected at 43.7 min was determined by mass spectrometry (MS). Fluorescence and electronic microscopy were used to evaluate the mechanism of action. Toxicity was tested on HeLa cells and human erythrocytes; physicochemical properties were evaluated. The molecule in the ES was characterized as sarconesin II and it showed activity against Gram-negative (Escherichia coli MG1655, Pseudomonas aeruginosa ATCC 27853, P. aeruginosa PA14) and Gram-positive (Staphylococcus aureus ATCC 29213, Micrococcus luteus A270) bacteria. The lowest minimum inhibitory concentration obtained was 1.9 μM for M. luteus A270; the AMP had no toxicity in any cells tested here and its action in bacterial membrane and DNA was confirmed. Sarconesin II was documented as a conserved domain of the ATP synthase protein belonging to the Fli-1 superfamily. The data reported here indicated that peptides could be alternative therapeutic candidates for use in infections against Gram-negative and Gram-positive bacteria and eventually as a new resource of compounds for combating multidrug-resistant bacteria.
Andrea Díaz-Roa; Abraham Espinoza-Culupú; Orlando Torres-García; Monamaris M. Borges; Ivan N. Avino; Flávio L. Alves; Antonio Miranda; Manuel A. Patarroyo; Jr. Pedro I. Da Silva; Felio J. Bello. Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions. Molecules 2019, 24, 2077 .
AMA StyleAndrea Díaz-Roa, Abraham Espinoza-Culupú, Orlando Torres-García, Monamaris M. Borges, Ivan N. Avino, Flávio L. Alves, Antonio Miranda, Manuel A. Patarroyo, Jr. Pedro I. Da Silva, Felio J. Bello. Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions. Molecules. 2019; 24 (11):2077.
Chicago/Turabian StyleAndrea Díaz-Roa; Abraham Espinoza-Culupú; Orlando Torres-García; Monamaris M. Borges; Ivan N. Avino; Flávio L. Alves; Antonio Miranda; Manuel A. Patarroyo; Jr. Pedro I. Da Silva; Felio J. Bello. 2019. "Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions." Molecules 24, no. 11: 2077.
Invasive Candida infections are an important growing medical concern and treatment options are limited to a few antifungal drug classes, with limited efficacies depending on the infecting organism. In this scenario, invasive infections caused by multiresistant Candida auris are emerging in several places around the world as important healthcare-associated infections. As antimicrobial peptides (AMPs) exert their activities primarily through mechanisms involving membrane disruption, they have a lower chance of inducing drug resistance than general chemical antimicrobials. Interestingly, we previously described the potent candicidal effect of a rattlesnake AMP, crotamine, against standard and treatment-resistant clinical isolates, with no hemolytic activity. We evaluated the antifungal susceptibility of several Candida spp. strains cultured from different patients by using the Clinical and Laboratory Standards Institute (CLSI) microdilution assay, and the antifungal activity of native crotamine was evaluated by a microbial growth inhibition microdilution assay. Although all Candida isolates evaluated here showed resistance to amphotericin B and fluconazole, crotamine (40–80 µM) exhibited in vitro activity against most isolates tested. We suggest that this native polypeptide from the South American rattlesnake Crotalus durissus terrificus has potential as a structural model for the generation of a new class of antimicrobial compounds with the power to fight against multiresistant Candida spp.
Caroline Dal Mas; Luana Rossato; Thaís Shimizu; Eduardo B. Oliveira; Pedro I. Da Silva Junior; Jacques F. Meis; Arnaldo Lopes Colombo; Mirian A. F. Hayashi. Effects of the Natural Peptide Crotamine from a South American Rattlesnake on Candida auris, an Emergent Multidrug Antifungal Resistant Human Pathogen. Biomolecules 2019, 9, 205 .
AMA StyleCaroline Dal Mas, Luana Rossato, Thaís Shimizu, Eduardo B. Oliveira, Pedro I. Da Silva Junior, Jacques F. Meis, Arnaldo Lopes Colombo, Mirian A. F. Hayashi. Effects of the Natural Peptide Crotamine from a South American Rattlesnake on Candida auris, an Emergent Multidrug Antifungal Resistant Human Pathogen. Biomolecules. 2019; 9 (6):205.
Chicago/Turabian StyleCaroline Dal Mas; Luana Rossato; Thaís Shimizu; Eduardo B. Oliveira; Pedro I. Da Silva Junior; Jacques F. Meis; Arnaldo Lopes Colombo; Mirian A. F. Hayashi. 2019. "Effects of the Natural Peptide Crotamine from a South American Rattlesnake on Candida auris, an Emergent Multidrug Antifungal Resistant Human Pathogen." Biomolecules 9, no. 6: 205.
The remarkable ability of microorganisms to develop resistance to conventional antibiotics is one of the biggest challenges that the pharmaceutical industry currently faces. Recent studies suggest that antimicrobial peptides discovered in spider venoms may be useful resources for the design of structurally new anti-infective agents effective against drug-resistant microorganisms. In this work, we found an anionic antibacterial peptide named U1-SCRTX-Lg1a in the venom of the spider Loxosceles gaucho. The peptide was purified using high-performance liquid chromatography (HPLC), its antimicrobial activity was tested through liquid growth inhibition assays, and its chemical properties were characterized using mass spectrometry. U1-SCRTX-Lg1a was found to show a monoisotopic mass of 1695.75 Da, activity against Gram-negative bacteria, a lack of hemolytic effects against human red blood cells, and a lack of cytotoxicity against human cervical carcinoma cells (HeLa). Besides this, the sequence of the peptide exhibited great similarity to specific regions of phospholipases D from different species of Loxosceles spiders, leading to the hypothesis that U1-SCRTX-Lg1a may have originated from a limited proteolytic cleavage. Our data suggest that U1-SCRTX-Lg1a is a promising candidate for the development of new antibiotics that could help fight bacterial infections and represents an exciting discovery for Loxosceles spiders.
Paula J Segura-Ramírez; Pedro I Silva Júnior. Loxosceles gaucho Spider Venom: An Untapped Source of Antimicrobial Agents. Toxins 2018, 10, 522 .
AMA StylePaula J Segura-Ramírez, Pedro I Silva Júnior. Loxosceles gaucho Spider Venom: An Untapped Source of Antimicrobial Agents. Toxins. 2018; 10 (12):522.
Chicago/Turabian StylePaula J Segura-Ramírez; Pedro I Silva Júnior. 2018. "Loxosceles gaucho Spider Venom: An Untapped Source of Antimicrobial Agents." Toxins 10, no. 12: 522.
Larval therapy (LT) is an alternative treatment for healing chronic wounds; its action is based on debridement, the removal of bacteria, and stimulating granulation tissue. The most important mechanism when using LT for combating infection depends on larval excretions and secretions (ES). Larvae are protected against infection by a spectrum of antimicrobial peptides (AMPs); special interest in AMPs has also risen regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during LT. Sarconesiopsis magellanica (Diptera: Calliphoridae) is a promising medically-important necrophagous fly. This article reports a small AMP being isolated from S. magellanica ES products for the first time; these products were obtained from third-instar larvae taken from a previously-established colony. ES were fractionated by RP-HPLC using C18 columns for the first analysis; the products were then lyophilised and their antimicrobial activity was characterized by incubation with different bacterial strains. These fractions’ primary sequences were determined by mass spectrometry and de novo sequencing; five AMPs were obtained, the Sarconesin fraction was characterized and antibacterial activity was tested in different concentrations with minimum inhibitory concentrations starting at 1.2 μM. Potent inhibitory activity was shown against Gram-negative (Escherichia coli D31, E. coli DH5α, Salmonella enterica ATCC 13314, Pseudomonas aeruginosa 27853) and Gram-positive (Staphylococcus aureus ATCC 29213, S. epidermidis ATCC 12228, Micrococcus luteus A270) bacteria. Sarconesin has a significant similarity with Rho-family GTPases which are important in organelle development, cytoskeletal dynamics, cell movement, and wound repair. The data reported here indicated that Sarconesin could be an alternative candidate for use in therapeutics against Gram-negative and Gram-positive bacterial infections. Our study describes one peptide responsible for antibacterial activity when LT is being used. The results shown here support carrying out further experiments aimed at validating S. magellanica AMPs as novel resources for combating antibacterial resistance.
Andrea Díaz-Roa; Manuel A. Patarroyo; Felio J. Bello; Pedro I. Jr. Da Silva. Sarconesin: Sarconesiopsis magellanica Blowfly Larval Excretions and Secretions With Antibacterial Properties. Frontiers in Microbiology 2018, 9, 2249 .
AMA StyleAndrea Díaz-Roa, Manuel A. Patarroyo, Felio J. Bello, Pedro I. Jr. Da Silva. Sarconesin: Sarconesiopsis magellanica Blowfly Larval Excretions and Secretions With Antibacterial Properties. Frontiers in Microbiology. 2018; 9 ():2249.
Chicago/Turabian StyleAndrea Díaz-Roa; Manuel A. Patarroyo; Felio J. Bello; Pedro I. Jr. Da Silva. 2018. "Sarconesin: Sarconesiopsis magellanica Blowfly Larval Excretions and Secretions With Antibacterial Properties." Frontiers in Microbiology 9, no. : 2249.
Purpose: Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian and mice neuromuscular preparation.
Sandro Rostelato-Ferreira; Cháriston André Dal Belo; Pedro Ismael Da Silva Junior; Stephen Hyslop; Léa Rodrigues-Simioni; Thomaz Augusto Alves Rocha-E-Silva. Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison. Advanced Pharmaceutical Bulletin 2018, 8, 517 -522.
AMA StyleSandro Rostelato-Ferreira, Cháriston André Dal Belo, Pedro Ismael Da Silva Junior, Stephen Hyslop, Léa Rodrigues-Simioni, Thomaz Augusto Alves Rocha-E-Silva. Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison. Advanced Pharmaceutical Bulletin. 2018; 8 (3):517-522.
Chicago/Turabian StyleSandro Rostelato-Ferreira; Cháriston André Dal Belo; Pedro Ismael Da Silva Junior; Stephen Hyslop; Léa Rodrigues-Simioni; Thomaz Augusto Alves Rocha-E-Silva. 2018. "Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison." Advanced Pharmaceutical Bulletin 8, no. 3: 517-522.
In contrast to vertebrate immune systems, invertebrates lack an adaptive response and rely solely on innate immunity in which antimicrobial peptides (AMPs) play an essential role. Most of them are membrane active molecules that are typically unstructured in solution and adopt secondary/tertiary structures upon binding to phospholipid bilayers. This work presents the first characterization of a constitutive AMP from the hemolymph of an Opiliones order animal: the harvestman Acutisoma longipes. This peptide was named longipin. It presents 18 aminoacid residues (SGYLPGKEYVYKYKGKVF) and a positive net charge at neutral pH. No similarity with other AMPs was observed. However, high sequence similarity with heme-lipoproteins from ticks suggested that longipin might be a protein fragment. The synthetic peptide showed enhanced antifungal activity against Candida guilliermondii and C. tropicalis yeasts (MIC: 3.8-7.5 μM) and did not interfered with VERO cells line viability at all concentrations tested (200-0.1 μM). This selectivity against microbial cells is related to the highest affinity of longipin for anionic charged vesicles (POPG:POPC) compared to zwitterionic ones (POPC), once microbial plasma membrane are generally more negatively charged compared to mammalian cells membrane. Dye leakage from carboxyfluorescein-loaded POPG:POPC vesicles suggested that longipin is a membrane active antimicrobial peptide and FT-IR spectroscopy showed that the peptide chain is mainly unstructured in solution or in the presence of POPC vesicles. However, upon binding to POPG:POPC vesicles, the FT-IR spectrum showed bands related to β-sheet and amyloid-like fibril conformations in agreement with thioflavin-T binding assays, indicating that longipin is an amyloid antimicrobial peptide.
Raphael Santa Rosa Sayegh; Isabel De Fátima Correia Batista; Robson Lopes De Melo; Karin A. Riske; Sirlei Daffre; Guillermo Montich; Pedro Ismael Da Silva Junior. Longipin: An Amyloid Antimicrobial Peptide from the Harvestman Acutisoma longipes (Arachnida: Opiliones) with Preferential Affinity for Anionic Vesicles. PLoS ONE 2016, 11, e0167953 .
AMA StyleRaphael Santa Rosa Sayegh, Isabel De Fátima Correia Batista, Robson Lopes De Melo, Karin A. Riske, Sirlei Daffre, Guillermo Montich, Pedro Ismael Da Silva Junior. Longipin: An Amyloid Antimicrobial Peptide from the Harvestman Acutisoma longipes (Arachnida: Opiliones) with Preferential Affinity for Anionic Vesicles. PLoS ONE. 2016; 11 (12):e0167953.
Chicago/Turabian StyleRaphael Santa Rosa Sayegh; Isabel De Fátima Correia Batista; Robson Lopes De Melo; Karin A. Riske; Sirlei Daffre; Guillermo Montich; Pedro Ismael Da Silva Junior. 2016. "Longipin: An Amyloid Antimicrobial Peptide from the Harvestman Acutisoma longipes (Arachnida: Opiliones) with Preferential Affinity for Anionic Vesicles." PLoS ONE 11, no. 12: e0167953.
We describe the mating behavior in the spermatheca-lacking theraphosid species Sickius longibulbi Soares & Camargo 1948. The behavior in captivity of nine pairs of S. longibulbi was videotaped and analyzed. The matting of this species presented an uncommon theraphosid pattern. There is little in the way of overt courtship by the male, the primary behavior seen being the male`s use of legs I and II to touch the female`s first pairs of legs and her chelicerae. Sometimes the male clasped one of the female`s first pairs of legs, bringing her close to him. While the female raised her body, the male clasped her fangs and held her tightly with his legs III wrapped around her prosoma. The male seemed to try to knock the female down, pushing her entire body until she lay on her dorsum. In this phase we observed the male biting the female on the sternum or on the leg joints. When the female fell, the male attempted to position himself at an angle of 90 degrees from the female. These movements appear to demand a lot of energy, particularly because the female is not passive during the mating. Our findings suggest that copulating in this position is, for the male, more successful than adopting other positions because it allows his extremely long palpal bulbs to deposit more sperm in the female oviduct where - since she lacks spermathecae - she retains the sperm. We suggest that the further he reaches into the oviduct, the greater the chance that he will fertilize the female`s eggs.FAPESP[03/12587-4]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP[06/58326-5]Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)CAPE
Rogério Bertani; Caroline Sayuri Fukushima; Pedro Ismael Da Silva Júnior. Mating behavior of Sickius longibulbi (Araneae, Theraphosidae, Ischnocolinae), a spider that lacks spermathecae. The Journal of Arachnology 2008, 36, 331 -335.
AMA StyleRogério Bertani, Caroline Sayuri Fukushima, Pedro Ismael Da Silva Júnior. Mating behavior of Sickius longibulbi (Araneae, Theraphosidae, Ischnocolinae), a spider that lacks spermathecae. The Journal of Arachnology. 2008; 36 (2):331-335.
Chicago/Turabian StyleRogério Bertani; Caroline Sayuri Fukushima; Pedro Ismael Da Silva Júnior. 2008. "Mating behavior of Sickius longibulbi (Araneae, Theraphosidae, Ischnocolinae), a spider that lacks spermathecae." The Journal of Arachnology 36, no. 2: 331-335.
The present study reports the identification of immune related transcripts from hemocytes of the spider Acanthoscurria gomesiana by high throughput sequencing of expressed sequence tags (ESTs). To generate ESTs from hemocytes, two cDNA libraries were prepared: one by directional cloning (primary) and the other by the normalization of the first (normalized). A total of 7584 clones were sequenced and the identical ESTs were clustered, resulting in 3723 assembled sequences (AS). At least 20% of these sequences are putative novel genes. The automatic functional annotation of AS based on Gene Ontology revealed several abundant transcripts related to the following functional classes: hemocyanin, lectin, and structural constituents of ribosome and cytoskeleton. From this annotation, 73 transcripts possibly involved in immune response were also identified, suggesting the existence of several molecular processes not previously described for spiders, such as: pathogen recognition, coagulation, complement activation, cell adhesion and intracellular signaling pathway for the activation of cellular defenses.
Daniel M. Lorenzini; Pedro I. Da Silva; Marcelo B. Soares; Paulo Arruda; João Setubal; Sirlei Daffre. Discovery of immune-related genes expressed in hemocytes of the tarantula spider Acanthoscurria gomesiana. Developmental & Comparative Immunology 2006, 30, 545 -556.
AMA StyleDaniel M. Lorenzini, Pedro I. Da Silva, Marcelo B. Soares, Paulo Arruda, João Setubal, Sirlei Daffre. Discovery of immune-related genes expressed in hemocytes of the tarantula spider Acanthoscurria gomesiana. Developmental & Comparative Immunology. 2006; 30 (6):545-556.
Chicago/Turabian StyleDaniel M. Lorenzini; Pedro I. Da Silva; Marcelo B. Soares; Paulo Arruda; João Setubal; Sirlei Daffre. 2006. "Discovery of immune-related genes expressed in hemocytes of the tarantula spider Acanthoscurria gomesiana." Developmental & Comparative Immunology 30, no. 6: 545-556.