Complete assembly, annotation of virulence genes and CRISPR editing of the genome of Leishmania amazonensis PH8 strain.

We report the sequencing and assembly of the PH8 strain of Leishmania amazonensis one of the etiological agents of leishmaniasis. After combining data from long Pacbio reads, short Illumina reads and synteny with the Leishmania mexicana genome, the sequence of 34 chromosomes with 8317 annotated genes was generated. Multigene families encoding three virulence factors, A2, amastins and the GP63 metalloproteases, were identified and compared to their annotation in other Leishmania species. As they have been recently recognized as virulence factors essential for disease establishment and progression of the infection, we also identified 14 genes encoding proteins involved in parasite iron and heme metabolism and compared to genes from other Trypanosomatids. To follow these studies with a genetic approach to address the role of virulence factors, we tested two CRISPR-Cas9 protocols to generate L. amazonensis knockout cell lines, using the Miltefosine transporter gene as a proof of concept.

Adhesive systems applied to dentin substrate under electric current: systematic review.

OBJECTIVES: The purpose of this systematic review was to collect and discuss the technique of adhesive systems application on dentin substrate under electric current. MATERIALS AND METHODS: The first search strategy was based on data available at PubMed, LILACS, Scielo, Scopus, and Cochrane Library, using a combination of descriptors such as "dentin bond agents OR adhesive system AND electric current OR electrobond" or "dentin bonding agents OR dentin bonding agent application OR adhesive system AND electric current OR electrobond", with no limit regarding the publication year. The second search strategy was based on the articles' references found previously. An additional search strategy was applied that concerned the proposed theme in the SBU-UNICAMP (Unicamp's Library System Institutional Repository). RESULTS: Twelve studies published between 2006 and 2020 were found. The analyses of the selected studies showed that the use of electric current during adhesive systems application on dentin, whether conventional or self-conditioning, increases resinous monomer infiltration in the dentin substrate, which improves the hybridization processes and the bond strength of the restorative material to dentin. CONCLUSIONS: Despite the favorable results related to the use of this technique, there is still no specific protocol for the application of adhesive systems under electric current.