Figure S1. DNA nucleotide sequence for thioredoxin and clavMo (underlined). Start codons of both ... more Figure S1. DNA nucleotide sequence for thioredoxin and clavMo (underlined). Start codons of both carrier and peptide sequence are indicated in bold. Stop codon is highlighted in red
Figure S3. Western blot analysis of the supernatant fraction of lysed E. coli strains. A) pLyzS/p... more Figure S3. Western blot analysis of the supernatant fraction of lysed E. coli strains. A) pLyzS/pET21-clavMO, B) pRIL/pET21-clavMO
Figure S4. Western blot analysis of the pellet cell fraction of lysed E. coli strains. A) pLyzS/p... more Figure S4. Western blot analysis of the pellet cell fraction of lysed E. coli strains. A) pLyzS/pET21-clavMO, B) pRIL/pET21-clavMO
Figure S2. A) Expression vector pET21a(+) used as backbone to insert the expression cassette; B) ... more Figure S2. A) Expression vector pET21a(+) used as backbone to insert the expression cassette; B) Expression cassette comprised of a signal sequence (Ss), a HIS-tag, the thioredoxin gene end the ClavMO peptide. In the white boxes are presented the restriction sites
Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcript... more Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcription-PCR from cotton boll weevil (Anthonomus grandis) larvae. A large diversity between the sequences was observed, with a mean pairwise identity of 22%. The cDNAs encompassed 11 trypsinlike sequences classifiable into three families and 3 chymotrypsin-like sequences belonging to a single family. Using a combination of 5 ´ and 3 ´ RACE the full length sequence was obtained for five of the cDNAs, named Agser2, Agser5, Agser6, Agser10 and Agser21. The encoded protein included amino acid sequence motifs of serine proteinase active sites, conserved cysteine residues, and both zymogen activation and signal peptides. Southern blotting analysis suggested that one or two copies of these serine genes existed in the A. grandis genome. Northern blotting analysis of Agser2 and Agser5 showed that for both genes expression is induced upon feeding and is concentrated in the gut of larvae and adult insec...
Objetivo principal: conocer las posibles influencias en la eleccion del tipo de parto, en la pers... more Objetivo principal: conocer las posibles influencias en la eleccion del tipo de parto, en la perspectiva de las mujeres embarazadas. Metodologia: se trata de un estudio cualitativo descriptivo realizado en dos unidades basicas de salud en un municipio en el sur de Brasil. El estudio incluyo a 12 mujeres embarazadas. La recoleccion de datos ocurrio entre agosto y octubre de 2012. Se utilizo la tecnica de analisis de contenido tematico. Resultados principales: el analisis de los temas surgio: la recuperacion del dolor y posparto; experiencias de parto; y profesionales de la salud. Conclusion principal: a la atencion prenatal, en preparacion para el parto tiene lagunas por cubrir por profesionales de la salud, para apoyar, responder preguntas y alentar un parto mas natural, para el bienestar de la madre y el nino par.
Journal of Tissue Engineering and Regenerative Medicine
Skin lesions are associated with functional/cosmetic problems for those afflicted. Scarless regen... more Skin lesions are associated with functional/cosmetic problems for those afflicted. Scarless regeneration is a challenge, not limited to the skin, and focus of active investigation. Recently, the host defense peptide innate defense regulatory peptide 1018 (IDR‐1018) has shown exciting regenerative properties. Nevertheless, literature regarding IDR‐1018 regenerative potential is scarce and limited to animal models. Here, we evaluated the regenerative potential of IDR‐1018 using human 2D and 3D human skin equivalents. First, we investigated IDR‐1018 using human cells found in skin—primary fibroblasts, primary keratinocytes, and the MeWo melanocytes cell line. IDR‐1018 promoted cell proliferation and expression of marker of proliferation Ki‐67, matrix metalloproteinase 1, and hyaluronan synthase 2 by fibroblasts. In keratinocytes, a drastic increase in expression was observed for Ki‐67, matrix metalloproteinase 1, C‐X‐C motif chemokine receptor type 4, C‐X‐C motif chemokine receptor type 7, fibroblast growth factor 2, hyaluronan synthase 2, vascular endothelial growth factor, and elastin, reflecting an intense stimulation of these cells. In melanocytes, increased migration and proliferation were observed following IDR‐1018 treatment. The capacity of IDR‐1018 to promote dermal contraction was verified using a dermal model. Finally, using a 3D human skin equivalent lesion model, we revealed that the regenerative potential of IDR1018, previously tested in mice and pigs, is valid for human skin tissue. Lesions closed faster in IDR‐1018‐treated samples, and the gene expression signature observed in 2D was reproduced in the 3D human skin equivalents. Overall, the present data show the regenerative potential of IDR‐1018 in an experimental system comprising human cells, underscoring the potential application for clinical investigation.
Cone snails are marine gastropod mollusks with one of the most powerful venoms in nature. The tox... more Cone snails are marine gastropod mollusks with one of the most powerful venoms in nature. The toxins, named conotoxins, must act quickly on the cone snails´ prey due to the fact that snails are extremely slow, reducing their hunting capability. Therefore, the characteristics of conotoxins have become the object of investigation, and as a result medicines have been developed or are in the trialing process. Conotoxins interact with transmembrane proteins, showing specificity and potency. They target ion channels and ionotropic receptors with greater regularity, and when interaction occurs, there is immediate physiological decompensation. In this review we aimed to evaluate the structural features of conotoxins and the relationship with their target types.
In the last years, the antimicrobial resistance against antibiotics has become a serious health i... more In the last years, the antimicrobial resistance against antibiotics has become a serious health issue, arise as global threat. This has generated a search for new strategies in the progress of new antimicrobial therapies. In this context, different nanosystems with antimicrobial properties have been studied. Specifically, magnetic nanoparticles seem to be very attractive due to their relatively simple synthesis, intrinsic antimicrobial activity, low toxicity and high versatility. Iron oxide NPs (IONPs) was authorized by the World Health Organization for human used in biomedical applications such as in vivo drug delivery systems, magnetic guided therapy and contrast agent for magnetic resonance imaging have been widely documented. Furthermore, the antimicrobial activity of different magnetic nanoparticles has recently been demonstrated. This review elucidates the recent progress of IONPs in drug delivery systems and focuses on the treatment of infectious diseases and target the possible detrimental biological effects and associated safety issues.
Among the numerous strategies plants have developed to fend off enemy attack, antimicrobial pepti... more Among the numerous strategies plants have developed to fend off enemy attack, antimicrobial peptides (AMPs) stand out as one of the most prominent defensive barriers that grant direct and durable resistance against a wide range of pests and pathogens. These small proteins are characterized by a compact structure and an overall positive charge. AMPs have an ancient origin and widespread occurrence in the plant kingdom but show an unusually high degree of variation in their amino acid sequences. Interestingly, there is a strikingly conserved topology among the plant AMP families, suggesting that the defensive properties of these peptides are not determined by their primary sequences but rather by their tridimensional structure. To explore and expand this idea, we here discuss the role of AMPs for plant defense from a structural perspective. We show how specific structural properties, such as length, charge, hydrophobicity, polar angle and conformation, are essential for plant AMPs to ...
Selective pressure imposed by millions of years of relentless biological attack has led to the de... more Selective pressure imposed by millions of years of relentless biological attack has led to the development of an extraordinary array of defense strategies in plants. Among these, antimicrobial peptides (AMPs) stand out as one of the most prominent components of the plant immune system. These small and usually basic peptides are deployed as a generalist defense strategy that grants direct and durable resistance against biotic stress. Even though their name implies a function against microbes, the range of plant-associated organisms affected by these peptides is much broader. In this review, we highlight the advances in our understanding on the role of AMPs in plant immunity. We demonstrate that the capacity of plant AMPs to act against a large spectrum of enemies relies on their diverse mechanism of action and remarkable structural stability. The efficacy of AMPs as a defense strategy is evidenced by their widespread occurrence in the plant kingdom, an astonishing heterogeneity in ho...
Antimicrobial peptides (AMPs) have attracted considerable attention because of their multiple and... more Antimicrobial peptides (AMPs) have attracted considerable attention because of their multiple and complex mechanisms of action toward resistant bacteria. However, reports have increasingly highlighted how bacteria can escape AMP administration. Here, the molecular mechanisms involved in Escherichia coli resistance to magainin I were investigated through comparative transcriptomics. Sub-inhibitory concentrations of magainin I were used to generate four experimental groups, including magainin I-susceptible E. coli, in the absence (C) and presence of magainin I (CM); and magainin I-resistant E. coli in the absence (R) and presence of magainin I (RM). The total RNA from each sample was extracted; cDNA libraries were constructed and further submitted for Illumina MiSeq sequencing. After RNA-seq data pre-processing and functional annotation, a total of 103 differentially expressed genes (DEGs) were identified, mainly related to bacterial metabolism. Moreover, down-regulation of cell motil...
Antimicrobial peptides (AMPs) are mostly endogenous, cationic, amphipathic polypeptides, produced... more Antimicrobial peptides (AMPs) are mostly endogenous, cationic, amphipathic polypeptides, produced by many natural sources. Recently, many biological functions beyond antimicrobial activity have been attributed to AMPs, and some of these have attracted the attention of the cosmetics industry. AMPs have revealed antioxidant, self-renewal and pro-collagen effects, which are desirable in anti-aging cosmetics. Additionally, AMPs may also be customized to act on specific cellular targets. Here, we review the recent literature that highlights the many possibilities presented by AMPs, focusing on the relevance and impact that this potentially novel class of active cosmetic ingredients might have in the near future, creating new market outlooks for the cosmetic industry with these molecules as a viable alternative to conventional cosmetics.
Streptococcus pyogenes, also known as group A Streptococcus (GAS), is a human pathogen that cause... more Streptococcus pyogenes, also known as group A Streptococcus (GAS), is a human pathogen that causes diverse human diseases including streptococcal toxic shock syndrome (STSS). A GAS outbreak occurred in Brasilia, Brazil, during the second half of the year 2011, causing 26 deaths. Whole genome sequencing was performed using Illumina platform. The sequences were assembled and genes were predicted for comparative analysis with emm type 1 strains: MGAS5005 and M1 GAS. Genomics comparison revealed one of the invasive strains that differ from others isolates and from emm 1 reference genomes. Also, the new invasive strain showed differences in the content of virulence factors compared to other isolated in the same outbreak. The evolution of contemporary GAS strains is strongly associated with horizontal gene transfer. This is the first genomic study of a Streptococcal emm 1 outbreak in Brazil, and revealed the rapid bacterial evolution leading to new clones. The emergence of new invasive st...
In recent years the antimicrobial peptides (AMPs) have been prospected and designed as new altern... more In recent years the antimicrobial peptides (AMPs) have been prospected and designed as new alternatives to conventional antibiotics. Indeed, AMPs have presented great potential toward pathogenic bacterial strains by means of complex mechanisms of action. However, reports have increasingly emerged regarding the mechanisms by which bacteria resist AMP administration. In this context, we performed a comparative proteomic study by using the total bacterial lysate of magainin I-susceptible and -resistant E. coli strains. After nanoUPLC-MS(E) analyses we identified 742 proteins distributed among the experimental groups, and 25 proteins were differentially expressed in the resistant strains. Among them 10 proteins involved in bacterial resistance, homeostasis, nutrition and protein transport were upregulated, while 15 proteins related to bacterial surface modifications, genetic information and β-lactams binding-protein were downregulated. Moreover, 60 exclusive proteins were identified in ...
Antimicrobial agents and chemotherapy, Aug 30, 2017
Fungal Candida species are commensals present in the mammalian skin and mucous membranes. Candida... more Fungal Candida species are commensals present in the mammalian skin and mucous membranes. Candida spp. are capable of breaching the epithelial barrier of immuno-compromised patients with neutrophil and cell-mediated immune dysfunctions, and it can also disseminate to multiple organs through the bloodstream. Here we examined the action of innate defence regulator (IDR)-1018, a 12-aminoacid residue peptide derived from bovine bactenecin (Bac2A), showed weak antifungal and anti-biofilm activity against C. albicans laboratory (ATCC 10231) and a clinical isolate (CI) strains (MICs of 32 μg.mL(-1) and 64 μg.mL(-1), respectively), while 8-fold lower concentrations led to dissolution of the fungal cells from pre-formed biofilms. IDR-1018 at 128 μg.mL(-1) was not hemolytic when tested against murine red blood cells, and also has not shown cytotoxic effect on murine monocyte RAW 264.7 and primary murine macrophage cells at the tested concentrations. IDR-1018 modulated the cytokine profile due...
The extraction and purification of parigidin-br3, a cyclotide analogue belonging to the "bra... more The extraction and purification of parigidin-br3, a cyclotide analogue belonging to the "bracelet" subfamily, from Palicourea rigida leaves is discussed. Unlike conventional cyclotides, parigidin-br3 has free N- and C-termini, as identified by MALDI-TOF/TOF analysis and confirmed by gene structure elucidation, and is one of a small number of acyclotides discovered during recent years. Parigidin-br3 showed cytotoxic activity against MCF-7 (breast cancer) and CACO2 (colorectal adenocarcinoma) cells, with IC50 values of ∼2.5 μM and less than 10% hemolytic activity. Overall, parigidin-br3 is a promising new molecule with cytotoxic properties against tumor cell lines and, unlike many synthetic acyclic analogues, demonstrates that cytotoxic activity is not limited to conventional (i.e., cyclic) cyclotides.
Anti-infective drugs have had a key role in the contemporary world, contributing to dramatically ... more Anti-infective drugs have had a key role in the contemporary world, contributing to dramatically decrease mortality rates caused by infectious diseases worldwide. Antimicrobial peptides (AMPs) are multifunctional effectors of the innate immune system of mucosal surfaces and present antimicrobial activity against a range of pathogenic viruses, bacteria, and fungi. However, the discovery and development of new antibacterial drugs is a crucial step to overcome the great challenge posed by the emergence of antibiotic resistance. In this review, we outline recent advances in the development of novel AMPs with improved antimicrobial activities that were achieved through characteristic structural design. In addition, we describe recent progress made to overcome some of the major limitations that have hindered peptide biosynthesis.
Cyclotides are multifunctional plant cyclic peptides containing 28-37 amino acid residues and a p... more Cyclotides are multifunctional plant cyclic peptides containing 28-37 amino acid residues and a pattern of three disulfide bridges, forming a motif known as the cyclic cystine knot. Due to their high biotechnological potential, the sequencing and characterization of cyclotide genes are crucial not only for cloning and establishing heterologous expression strategies, but also to understand local plant evolution in the context of host-pathogen relationships. Here, two species from the Brazilian Cerrado, Palicourea rigida (Rubiaceae) and Pombalia lanata (A.St.-Hil.) Paula-Souza (Violaceae), were used for cloning and characterizing novel cyclotide genes. Using 3' and 5' RACE PCR and sequencing, two full cDNAs, named parigidin-br2 (P. rigida) and hyla-br1 (P. lanata), were isolated and shown to have similar genetic structures to other cyclotides. Both contained the conserved ER-signal domain, N-terminal prodomain, mature cyclotide domain and a C-terminal region. Genomic sequencin...
Figure S1. DNA nucleotide sequence for thioredoxin and clavMo (underlined). Start codons of both ... more Figure S1. DNA nucleotide sequence for thioredoxin and clavMo (underlined). Start codons of both carrier and peptide sequence are indicated in bold. Stop codon is highlighted in red
Figure S3. Western blot analysis of the supernatant fraction of lysed E. coli strains. A) pLyzS/p... more Figure S3. Western blot analysis of the supernatant fraction of lysed E. coli strains. A) pLyzS/pET21-clavMO, B) pRIL/pET21-clavMO
Figure S4. Western blot analysis of the pellet cell fraction of lysed E. coli strains. A) pLyzS/p... more Figure S4. Western blot analysis of the pellet cell fraction of lysed E. coli strains. A) pLyzS/pET21-clavMO, B) pRIL/pET21-clavMO
Figure S2. A) Expression vector pET21a(+) used as backbone to insert the expression cassette; B) ... more Figure S2. A) Expression vector pET21a(+) used as backbone to insert the expression cassette; B) Expression cassette comprised of a signal sequence (Ss), a HIS-tag, the thioredoxin gene end the ClavMO peptide. In the white boxes are presented the restriction sites
Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcript... more Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcription-PCR from cotton boll weevil (Anthonomus grandis) larvae. A large diversity between the sequences was observed, with a mean pairwise identity of 22%. The cDNAs encompassed 11 trypsinlike sequences classifiable into three families and 3 chymotrypsin-like sequences belonging to a single family. Using a combination of 5 ´ and 3 ´ RACE the full length sequence was obtained for five of the cDNAs, named Agser2, Agser5, Agser6, Agser10 and Agser21. The encoded protein included amino acid sequence motifs of serine proteinase active sites, conserved cysteine residues, and both zymogen activation and signal peptides. Southern blotting analysis suggested that one or two copies of these serine genes existed in the A. grandis genome. Northern blotting analysis of Agser2 and Agser5 showed that for both genes expression is induced upon feeding and is concentrated in the gut of larvae and adult insec...
Objetivo principal: conocer las posibles influencias en la eleccion del tipo de parto, en la pers... more Objetivo principal: conocer las posibles influencias en la eleccion del tipo de parto, en la perspectiva de las mujeres embarazadas. Metodologia: se trata de un estudio cualitativo descriptivo realizado en dos unidades basicas de salud en un municipio en el sur de Brasil. El estudio incluyo a 12 mujeres embarazadas. La recoleccion de datos ocurrio entre agosto y octubre de 2012. Se utilizo la tecnica de analisis de contenido tematico. Resultados principales: el analisis de los temas surgio: la recuperacion del dolor y posparto; experiencias de parto; y profesionales de la salud. Conclusion principal: a la atencion prenatal, en preparacion para el parto tiene lagunas por cubrir por profesionales de la salud, para apoyar, responder preguntas y alentar un parto mas natural, para el bienestar de la madre y el nino par.
Journal of Tissue Engineering and Regenerative Medicine
Skin lesions are associated with functional/cosmetic problems for those afflicted. Scarless regen... more Skin lesions are associated with functional/cosmetic problems for those afflicted. Scarless regeneration is a challenge, not limited to the skin, and focus of active investigation. Recently, the host defense peptide innate defense regulatory peptide 1018 (IDR‐1018) has shown exciting regenerative properties. Nevertheless, literature regarding IDR‐1018 regenerative potential is scarce and limited to animal models. Here, we evaluated the regenerative potential of IDR‐1018 using human 2D and 3D human skin equivalents. First, we investigated IDR‐1018 using human cells found in skin—primary fibroblasts, primary keratinocytes, and the MeWo melanocytes cell line. IDR‐1018 promoted cell proliferation and expression of marker of proliferation Ki‐67, matrix metalloproteinase 1, and hyaluronan synthase 2 by fibroblasts. In keratinocytes, a drastic increase in expression was observed for Ki‐67, matrix metalloproteinase 1, C‐X‐C motif chemokine receptor type 4, C‐X‐C motif chemokine receptor type 7, fibroblast growth factor 2, hyaluronan synthase 2, vascular endothelial growth factor, and elastin, reflecting an intense stimulation of these cells. In melanocytes, increased migration and proliferation were observed following IDR‐1018 treatment. The capacity of IDR‐1018 to promote dermal contraction was verified using a dermal model. Finally, using a 3D human skin equivalent lesion model, we revealed that the regenerative potential of IDR1018, previously tested in mice and pigs, is valid for human skin tissue. Lesions closed faster in IDR‐1018‐treated samples, and the gene expression signature observed in 2D was reproduced in the 3D human skin equivalents. Overall, the present data show the regenerative potential of IDR‐1018 in an experimental system comprising human cells, underscoring the potential application for clinical investigation.
Cone snails are marine gastropod mollusks with one of the most powerful venoms in nature. The tox... more Cone snails are marine gastropod mollusks with one of the most powerful venoms in nature. The toxins, named conotoxins, must act quickly on the cone snails´ prey due to the fact that snails are extremely slow, reducing their hunting capability. Therefore, the characteristics of conotoxins have become the object of investigation, and as a result medicines have been developed or are in the trialing process. Conotoxins interact with transmembrane proteins, showing specificity and potency. They target ion channels and ionotropic receptors with greater regularity, and when interaction occurs, there is immediate physiological decompensation. In this review we aimed to evaluate the structural features of conotoxins and the relationship with their target types.
In the last years, the antimicrobial resistance against antibiotics has become a serious health i... more In the last years, the antimicrobial resistance against antibiotics has become a serious health issue, arise as global threat. This has generated a search for new strategies in the progress of new antimicrobial therapies. In this context, different nanosystems with antimicrobial properties have been studied. Specifically, magnetic nanoparticles seem to be very attractive due to their relatively simple synthesis, intrinsic antimicrobial activity, low toxicity and high versatility. Iron oxide NPs (IONPs) was authorized by the World Health Organization for human used in biomedical applications such as in vivo drug delivery systems, magnetic guided therapy and contrast agent for magnetic resonance imaging have been widely documented. Furthermore, the antimicrobial activity of different magnetic nanoparticles has recently been demonstrated. This review elucidates the recent progress of IONPs in drug delivery systems and focuses on the treatment of infectious diseases and target the possible detrimental biological effects and associated safety issues.
Among the numerous strategies plants have developed to fend off enemy attack, antimicrobial pepti... more Among the numerous strategies plants have developed to fend off enemy attack, antimicrobial peptides (AMPs) stand out as one of the most prominent defensive barriers that grant direct and durable resistance against a wide range of pests and pathogens. These small proteins are characterized by a compact structure and an overall positive charge. AMPs have an ancient origin and widespread occurrence in the plant kingdom but show an unusually high degree of variation in their amino acid sequences. Interestingly, there is a strikingly conserved topology among the plant AMP families, suggesting that the defensive properties of these peptides are not determined by their primary sequences but rather by their tridimensional structure. To explore and expand this idea, we here discuss the role of AMPs for plant defense from a structural perspective. We show how specific structural properties, such as length, charge, hydrophobicity, polar angle and conformation, are essential for plant AMPs to ...
Selective pressure imposed by millions of years of relentless biological attack has led to the de... more Selective pressure imposed by millions of years of relentless biological attack has led to the development of an extraordinary array of defense strategies in plants. Among these, antimicrobial peptides (AMPs) stand out as one of the most prominent components of the plant immune system. These small and usually basic peptides are deployed as a generalist defense strategy that grants direct and durable resistance against biotic stress. Even though their name implies a function against microbes, the range of plant-associated organisms affected by these peptides is much broader. In this review, we highlight the advances in our understanding on the role of AMPs in plant immunity. We demonstrate that the capacity of plant AMPs to act against a large spectrum of enemies relies on their diverse mechanism of action and remarkable structural stability. The efficacy of AMPs as a defense strategy is evidenced by their widespread occurrence in the plant kingdom, an astonishing heterogeneity in ho...
Antimicrobial peptides (AMPs) have attracted considerable attention because of their multiple and... more Antimicrobial peptides (AMPs) have attracted considerable attention because of their multiple and complex mechanisms of action toward resistant bacteria. However, reports have increasingly highlighted how bacteria can escape AMP administration. Here, the molecular mechanisms involved in Escherichia coli resistance to magainin I were investigated through comparative transcriptomics. Sub-inhibitory concentrations of magainin I were used to generate four experimental groups, including magainin I-susceptible E. coli, in the absence (C) and presence of magainin I (CM); and magainin I-resistant E. coli in the absence (R) and presence of magainin I (RM). The total RNA from each sample was extracted; cDNA libraries were constructed and further submitted for Illumina MiSeq sequencing. After RNA-seq data pre-processing and functional annotation, a total of 103 differentially expressed genes (DEGs) were identified, mainly related to bacterial metabolism. Moreover, down-regulation of cell motil...
Antimicrobial peptides (AMPs) are mostly endogenous, cationic, amphipathic polypeptides, produced... more Antimicrobial peptides (AMPs) are mostly endogenous, cationic, amphipathic polypeptides, produced by many natural sources. Recently, many biological functions beyond antimicrobial activity have been attributed to AMPs, and some of these have attracted the attention of the cosmetics industry. AMPs have revealed antioxidant, self-renewal and pro-collagen effects, which are desirable in anti-aging cosmetics. Additionally, AMPs may also be customized to act on specific cellular targets. Here, we review the recent literature that highlights the many possibilities presented by AMPs, focusing on the relevance and impact that this potentially novel class of active cosmetic ingredients might have in the near future, creating new market outlooks for the cosmetic industry with these molecules as a viable alternative to conventional cosmetics.
Streptococcus pyogenes, also known as group A Streptococcus (GAS), is a human pathogen that cause... more Streptococcus pyogenes, also known as group A Streptococcus (GAS), is a human pathogen that causes diverse human diseases including streptococcal toxic shock syndrome (STSS). A GAS outbreak occurred in Brasilia, Brazil, during the second half of the year 2011, causing 26 deaths. Whole genome sequencing was performed using Illumina platform. The sequences were assembled and genes were predicted for comparative analysis with emm type 1 strains: MGAS5005 and M1 GAS. Genomics comparison revealed one of the invasive strains that differ from others isolates and from emm 1 reference genomes. Also, the new invasive strain showed differences in the content of virulence factors compared to other isolated in the same outbreak. The evolution of contemporary GAS strains is strongly associated with horizontal gene transfer. This is the first genomic study of a Streptococcal emm 1 outbreak in Brazil, and revealed the rapid bacterial evolution leading to new clones. The emergence of new invasive st...
In recent years the antimicrobial peptides (AMPs) have been prospected and designed as new altern... more In recent years the antimicrobial peptides (AMPs) have been prospected and designed as new alternatives to conventional antibiotics. Indeed, AMPs have presented great potential toward pathogenic bacterial strains by means of complex mechanisms of action. However, reports have increasingly emerged regarding the mechanisms by which bacteria resist AMP administration. In this context, we performed a comparative proteomic study by using the total bacterial lysate of magainin I-susceptible and -resistant E. coli strains. After nanoUPLC-MS(E) analyses we identified 742 proteins distributed among the experimental groups, and 25 proteins were differentially expressed in the resistant strains. Among them 10 proteins involved in bacterial resistance, homeostasis, nutrition and protein transport were upregulated, while 15 proteins related to bacterial surface modifications, genetic information and β-lactams binding-protein were downregulated. Moreover, 60 exclusive proteins were identified in ...
Antimicrobial agents and chemotherapy, Aug 30, 2017
Fungal Candida species are commensals present in the mammalian skin and mucous membranes. Candida... more Fungal Candida species are commensals present in the mammalian skin and mucous membranes. Candida spp. are capable of breaching the epithelial barrier of immuno-compromised patients with neutrophil and cell-mediated immune dysfunctions, and it can also disseminate to multiple organs through the bloodstream. Here we examined the action of innate defence regulator (IDR)-1018, a 12-aminoacid residue peptide derived from bovine bactenecin (Bac2A), showed weak antifungal and anti-biofilm activity against C. albicans laboratory (ATCC 10231) and a clinical isolate (CI) strains (MICs of 32 μg.mL(-1) and 64 μg.mL(-1), respectively), while 8-fold lower concentrations led to dissolution of the fungal cells from pre-formed biofilms. IDR-1018 at 128 μg.mL(-1) was not hemolytic when tested against murine red blood cells, and also has not shown cytotoxic effect on murine monocyte RAW 264.7 and primary murine macrophage cells at the tested concentrations. IDR-1018 modulated the cytokine profile due...
The extraction and purification of parigidin-br3, a cyclotide analogue belonging to the "bra... more The extraction and purification of parigidin-br3, a cyclotide analogue belonging to the "bracelet" subfamily, from Palicourea rigida leaves is discussed. Unlike conventional cyclotides, parigidin-br3 has free N- and C-termini, as identified by MALDI-TOF/TOF analysis and confirmed by gene structure elucidation, and is one of a small number of acyclotides discovered during recent years. Parigidin-br3 showed cytotoxic activity against MCF-7 (breast cancer) and CACO2 (colorectal adenocarcinoma) cells, with IC50 values of ∼2.5 μM and less than 10% hemolytic activity. Overall, parigidin-br3 is a promising new molecule with cytotoxic properties against tumor cell lines and, unlike many synthetic acyclic analogues, demonstrates that cytotoxic activity is not limited to conventional (i.e., cyclic) cyclotides.
Anti-infective drugs have had a key role in the contemporary world, contributing to dramatically ... more Anti-infective drugs have had a key role in the contemporary world, contributing to dramatically decrease mortality rates caused by infectious diseases worldwide. Antimicrobial peptides (AMPs) are multifunctional effectors of the innate immune system of mucosal surfaces and present antimicrobial activity against a range of pathogenic viruses, bacteria, and fungi. However, the discovery and development of new antibacterial drugs is a crucial step to overcome the great challenge posed by the emergence of antibiotic resistance. In this review, we outline recent advances in the development of novel AMPs with improved antimicrobial activities that were achieved through characteristic structural design. In addition, we describe recent progress made to overcome some of the major limitations that have hindered peptide biosynthesis.
Cyclotides are multifunctional plant cyclic peptides containing 28-37 amino acid residues and a p... more Cyclotides are multifunctional plant cyclic peptides containing 28-37 amino acid residues and a pattern of three disulfide bridges, forming a motif known as the cyclic cystine knot. Due to their high biotechnological potential, the sequencing and characterization of cyclotide genes are crucial not only for cloning and establishing heterologous expression strategies, but also to understand local plant evolution in the context of host-pathogen relationships. Here, two species from the Brazilian Cerrado, Palicourea rigida (Rubiaceae) and Pombalia lanata (A.St.-Hil.) Paula-Souza (Violaceae), were used for cloning and characterizing novel cyclotide genes. Using 3' and 5' RACE PCR and sequencing, two full cDNAs, named parigidin-br2 (P. rigida) and hyla-br1 (P. lanata), were isolated and shown to have similar genetic structures to other cyclotides. Both contained the conserved ER-signal domain, N-terminal prodomain, mature cyclotide domain and a C-terminal region. Genomic sequencin...
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