RESUMO
BACKGROUND: Bartonella bacilliformis is the causative agent of Carrion's disease, a neglected illness with mortality rates of 40-85% in the absence of treatment. The lack of a diagnostic technique to overcome misdiagnosis and treat asymptomatic carriers is of note. This study aimed to identify new B. bacilliformis antigenic candidates that could lead to a new diagnostic tool able to be implemented in endemic rural areas. METHODOLOGY/PRINCIPAL FINDINGS: Blood (n = 198) and serum (n = 177) samples were collected in northern Peru. Clinical data were recorded. Specific 16S rRNA amplification by RT-PCR, IFA and ELISA for IgM/IgG with whole cells as antigens was done. Western blot analysis and N-terminal amino acid sequencing detected seroreactive proteins. ELISAs for IgM/IgG for the antigenic candidates were performed. Of the population 33.3% reported at least one symptom compatible with Carrion's disease; 25.4% (IFA), 27.1% (ELISA-IgG), 33.9% (ELISA-IgM) and 38.9% (RT-PCR) of samples were positive. Four proteins were considered potential antigenic candidates, including two new antigenic candidates, succinyl-CoA synthetase subunit α (SCS-α) and succinyl-CoA synthetase subunit ß (SCS-ß). On Western blot both Pap31 and SCS-α interacted with IgM, while GroEL and SCS-ß interacted with IgG. The presence of specific antibodies against the antigenic candidates varied from 34.5% (IgG against SCS-α) to 97.2% (IgM against Pap31). CONCLUSIONS/SIGNIFICANCE: RT-PCR and the high levels of positivity for specific ELISAs demonstrate high levels of B. bacilliformis exposure and asymptomatic carriers among inhabitants. The new antigens identified might be used as a new rapid diagnostic tool to diagnose acute Carrion's disease and identify asymptomatic carriers.
Assuntos
Antígenos de Bactérias/imunologia , Infecções por Bartonella/microbiologia , Bartonella bacilliformis/imunologia , Succinato-CoA Ligases/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Infecções por Bartonella/imunologia , Western Blotting , Criança , Pré-Escolar , Chlorocebus aethiops , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Peru , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase em Tempo Real , Succinato-CoA Ligases/genética , Células Vero , Adulto JovemRESUMO
MicroRNAs (miRNAs) are non-coding small RNAs that regulate gene expression by translational repression or transcript degradation. A large number of miRNAs have been identified from model plant species; however, the character of conserved miRNAs is poorly understood. We studied 42 miRNA families that are conserved within the plant kingdom, using the miRBase database. Some conserved miRNA families were found to be preferentially expressed in dicots relative to monocots, especially miR403, miR472 and miR479. Using an improved homology search-based approach and the conserved miRNAs as the query set, 34 conserved miRNAs and the miR482 family were identified in wheat. Forty-six wheat mRNAs were predicted as their putative target genes. Most conserved wheat miRNAs were found to retain homologous target interactions and have analogous molecular functions. The miR172 displayed a wheat-specific function and was found to have an additional target interaction with succinyl-CoA ligase. We concluded that although miRNAs are conserved, the expression and function of some have drifted during long periods of plant evolution.
Assuntos
MicroRNAs/genética , Triticum/genética , Sequência de Bases , Sequência Conservada , DNA de Plantas , Evolução Molecular , Genes de Plantas , Técnicas Genéticas , Genômica , MicroRNAs/metabolismo , Modelos Genéticos , Dados de Sequência Molecular , RNA Mensageiro/metabolismo , Succinato-CoA Ligases/genéticaRESUMO
3' Untranslated region processing and polyadenylation in Trichomonas vaginalis was analyzed by 3' rapid amplification of cDNA ends and sequence analysis of T. vaginalis mRNAs. A putative polyadenylation signal with the sequence UAAA was found 11-30 nucleotides upstream from the cleavage site. The motif pyrimidine( downward arrow)(A)(0-3)AAUU is proposed to be the cleavage site for polyadenylation of transcripts. This potential sequence defining the cleavage site for polyadenylation in eukaryotes is a novel finding. As in other eukaryotes, runs of several U's downstream from the cleavage site were identified. A working hypothesis is proposed which couples the UAA translation stop codon with the signaling for the 3'end processing of transcripts in this early divergent parasitic protozoa.