Alfonso Gala-Garcia

The Corynebacterium pseudotuberculosis in silico predicted pan-exoproteome

BackgroundPan-genomic studies aim, for instance, at defining the core, dispensable and unique genes within a species. A pan-genomics study for vaccine design tries to assess the best candidates for a vaccine against a specific pathogen. In this context, rather than studying genes predicted to be exported in a single genome, with pan-genomics it is possible to study genes present in different strains within the same species, such as virulence factors. The target organism of this pan-genomic work here presented is Corynebacterium pseudotuberculosis, the etiologic agent of caseous lymphadenitis (CLA) in goat and sheep, which causes significant economic losses in those herds around the world. Currently, only a few antigens against CLA are known as being the basis of commercial and still ineffective vaccines. In this regard, the here presented work analyses, in silico, five C. pseudotuberculosis genomes and gathers data to predict common exported proteins in all five genomes. These candidates were also compared to two recent C. pseudotuberculosis in vitro exoproteome results.ResultsThe complete genome of five C. pseudotuberculosis strains (1002, C231, I19, FRC41 and PAT10) were submitted to pan-genomics analysis, yielding 306, 59 and 12 gene sets, respectively, representing the core, dispensable and unique in silico predicted exported pan-genomes. These sets bear 150 genes classified as secreted (SEC) and 227 as potentially surface exposed (PSE). Our findings suggest that the main C. pseudotuberculosis in vitro exoproteome could be greater, appended by a fraction of the 35 proteins formerly predicted as making part of the variant in vitro exoproteome. These genomes were manually curated for correct methionine initiation and redeposited with a total of 1885 homogenized genes.ConclusionsThe in silico prediction of exported proteins has allowed to define a list of putative vaccine candidate genes present in all five complete C. pseudotuberculosis genomes. Moreover, it has also been possible to define the in silico predicted dispensable and unique C. pseudotuberculosis exported proteins. These results provide in silico evidence to further guide experiments in the areas of vaccines, diagnosis and drugs. The work here presented is the first whole C. pseudotuberculosis in silico predicted pan-exoproteome completed till today.

Progression of ‘OMICS’ methodologies for understanding the pathogenicity of Corynebacterium pseudotuberculosis: the Brazilian experience

Since the first successful attempt at sequencing the Corynebacterium pseudotuberculosis genome, large amounts of genomic, transcriptomic and proteomic data have been generated. C. pseudotuberculosis is an interesting bacterium due to its great zoonotic potential and because it causes considerable economic losses worldwide. Furthermore, different strains of C. pseudotuberculosis are capable of causing various diseases in different hosts. Currently, we seek information about the phylogenetic relationships between different strains of C. pseudotuberculosis isolates from different hosts across the world and to employ these data to develop tools to diagnose and eradicate the diseases these strains cause. In this review, we present the latest findings on C. pseudotuberculosis that have been obtained with the most advanced techniques for sequencing and genomic organization. We also discuss the development of in silico tools for processing these data to prompt a better understanding of this pathogen.

Bioceramic/Poly (glycolic)-poly (lactic acid) composite induces mineralized barrier after direct capping of rat tooth pulp tissue

The aim of this study was to observe the histopathological pulp response following direct pulp capping of mechanically exposed teeth in rats with a composite of beta-tricalcium phosphate-hydroxyapatite bioceramic (BC) and poly (glycolic)-poly (lactic acid) (PLGA) material or a calcium hydroxide [Ca(OH)2] material, compared to BC alone and a negative control of water. Pulp of the maxillary molars was exposed, followed by capping with the experimental material. The pulpal tissue response was assessed post-operatively at 1, 7, 14 and 30 d, followed by histological analysis. The Ca(OH)2 group exhibited severe acute inflammatory cell infiltration at day 14. However after 30 d, a new hard tissue with macro porous obliteration of the pulp chamber and a characteristic necrotic area had appeared. BC and Ca(OH)2 capping were associated with moderate inflammation and dentinal bridge similar. Meanwhile, in the BC/PLGA composite group, there was moderate inflammatory infiltrate and formation of a dense and complete dentinal bridge. In conclusion, the BC/PLGA composite material showed a large zone of tertiary dentin, and effectively reorganized the dentin-pulp complex.

Effect of Aloe vera. on Rat Pulp Tissue

Abstract Aloe vera. L. (Aloeaceae) has been extensively studied for anti-inflammatory, antimicrobial, and cellular regeneration properties. This work evaluated in vivo. the effects of powder of freeze-dried Aloe vera. on rat pulp tissue. Pulp tissue was mechanically exposed to Aloe vera., and it was evaluated at 1 to 30 days after the procedure by histopathologic examination. A predominant acute-moderate inflammatory infiltrate was observed in the Aloe vera.–treated group 1 to 7 days after treatment. At 14 to 30 days, pulp tissue took a normal pattern when comparing Aloe vera. treatment with the Ca(OH)2 treatment; it was found that both treatment groups stimulated reparative dentin and the formation of complete bridge. Strong superficial necrosis was detected exclusively for Ca(OH)2. It seems evident that application of Aloe vera. in direct contact with the exposed pulp has acceptable biocompatibility and can lead to tertiary bridge formation.

Hybrid nanofibers based on poly-caprolactone/gelatin/hydroxyapatite nanoparticles-loaded Doxycycline: Effective anti-tumoral and antibacterial activity

Cancer is one of the leading causes of morbidity and mortality Worldwide, 19.3 million new cancer cases are expected to be identified in 2025. Among the therapeutic arsenal to cancer control one could find the Doxycycline and the nano hydroxyapatite. The Doxycycline (Dox) not only shown antibiotic effect but also exhibits a wide range of pleiotropic therapeutic properties as the control of the invasive and metastatic cancer cells characteristics. The purpose of the present study was to evaluate both cytotoxicity in vitro and antibacterial activity of electrospun Dox-loaded hybrid nanofibrous scaffolds composed by hydroxyapatite nanoparticles (nHA), poly-ε-caprolactone (PCL) and gelatin (Gel) polymers. Both nHA and Dox were dispersed into different PCL/Gel ratios (70:30, 60:40, 50:50wt%) solutions to form electrospun nanofibers. The nHA and Dox/nHA/PCL-Gel hybrid nanofibers were characterized by TEM microscopy. In vitro Dox release behavior from all of these Dox-loaded nHA/PCL-Gel nanofibers showed the same burst release profile due to the high solubility of Gel in the release medium. Antibacterial properties of nanofiber composites were evaluated using Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Porphyromonas gingivalis (P. gingivalis) bacteria. The co-delivery of nHA particles and Dox simultaneously exhibited inhibition of bacterial growth more efficiently than the delivery of either Dox or nHA at the same concentrations, indicating a synergistic effect. The results showed that cancer cell tested had different sensibility to co-delivery system. On the whole, A-431 cells were found exhibited the most pronounced synergistic effect compared to CACO-2 and 4T1 cancer cells. Based on the anticancer as well as the antimicrobial results in this study, the developed Dox/nHA/PCL-Gel composite nanofibers are suitable as a drug delivery system with potential applications in the biomedical fields.

Efficacy of coral-hydroxyapatite and biphasic calcium phosphate for early bacterial detection

Nano- or microhydroxyapatites with microbiological properties are being used to detect pathogens in clinical samples and industrial environments. In this study, the calcium phosphates coral-hydroxyapatite and biphasic calcium phosphate were characterized physicochemically using x-ray diffraction, thermogravimetric, and differential thermal analysis. The morphology, texture, and chemical composition of the ceramics were also investigated using scanning electron microscopy with energy dispersive spectroscopy. The biocompatibility of the ceramics was evaluated using Escherichia coli and Enterococcus faecalis. Microorganisms were detected by incorporating the enzyme markers 4-metilumbelliferil-β-d-glucoside and 4-metilumbelliferil-β-d-glucuronide in the ceramic powders and evaluating fluorescence. The characterization of the ceramics revealed typical characteristics, such as crystallinity, thermal stability, and chemical composition, consistent with other calcium phosphates. The calcium phosphates coral-hydroxyapatite and biphasic calcium phosphate ceramics differed from one another in morphology, structural topography, particle size distribution, and the capacity to absorb water. These properties can influence the rates of microbiological responses and bacterial detection. Although both materials are suitable for use as structural supports in microbial diagnostic systems, BCP was more efficient and detected E. coli and E. faecalis more rapidly than CHA.

Complete Genome Sequence of Corynebacterium pseudotuberculosis Strain E19, Isolated from a Horse in Chile

ABSTRACT Corynebacterium pseudotuberculosis is related to several diseases infecting horses and small ruminants, causing economic losses to agribusiness. Here, we present the genome sequence of C. pseudotuberculosis strain E19. The genome includes one circular chromosome 2,367,956 bp (52.1% G+C content), with 2,112 genes predicted, 12 rRNAs, and 48 tRNAs.