Carga proviral do HTLV-1 e HTLV-2: um método simples através da PCR quantitativa em tempo real HTLV-1 and HTLV-2 proviral load: a simple method using quantitative real-time PCR

Os vírus linfotrópicos de células T humanas, quando integrados ao genoma da célula hospedeira, provírus, têm como marcador de replicação seu DNA proviral. A carga proviral parece ser um importante fator no desenvolvimento de patologias associadas a estes retrovírus. Neste estudo foi desenvolvida uma metodologia para quantificação absoluta da carga proviral dos HTLV-1 e HTLV-2 através da PCR em tempo real. Cinqüenta e três amostras de doadores de sangue com teste de ELISA reagente foram submetidas à metodologia, que utilizou o sistema TaqMan® para três seqüências alvo: HTLV-1, HTLV-2 e albumina. A quantificação proviral absoluta foi determinada através da proporção relativa entre o genoma do HTLV e o genoma da célula hospedeira, levando em consideração o número de leucócitos. O método apresentado é sensível (215 cópias/mL), prático e simples para quantificação proviral, além de eficiente e adequado para confirmação e discriminação da infecção pelos tipos virais.<br>When the human T cell lymphotropic virus (HTLV) is integrated with the host cell genome (provirus), its proviral DNA is a replication marker. Proviral load appears to be an important factor in the development of diseases related to these retroviruses. In this study, a methodology for absolute quantification of the HTLV-1 and HTLV-2 proviral load using real-time PCR was developed. Fifty-three blood donor samples with positive ELISA test result were subjected to this methodology, which utilized the TaqMan® system for three target sequences: HTLV-1, HTLV-2 and albumin. The absolute proviral load was quantified using the relative ratio between the HTLV genome and the host cell genome, taking into consideration the white blood cell count. The method presented is sensitive (215 copies/ml), practical and simple for proviral quantification, and is efficient and appropriate for confirming and discriminating infections according to viral type

Neurological symptoms and signs in HTLV-1 patients with overactive bladder syndrome

OBJECTIVE: To compare neurological symptoms and signs in HTLV-1 asymptomatic carriers and HTLV-1 patients with overactive bladder (OB) syndrome. METHODS: We studied 102 HTLV-1 positive individuals without HAM/TSP (HTLV-1 associated myelopathy/tropical spastic paraparesis) divided into two groups according to the presence or absence of OB syndrome. Clinical interview, neurological exam and proviral load was performed in all patients. RESULTS AND CONCLUSIONS: Individuals with OB were more commonly female (84.3 vs. 60.8% of asymptomatics, p=0.01). The prevalence of neurological complaints was higher in OB group, especially hand or foot numbness and arm or leg weakness. There was no difference between the groups in neurological strength and reflexes. Weakness complaint remained strongly associated with OB in multivariate logistic regression analysis adjusting for sex and age [adjusted odds ratio and 95%CI 3.59 (1.45-8.88) in arms and 6.68 (2.63-16.93) in legs]. Proviral load was also different between the two groups with higher level on OB individuals

Phylogenetic analysis of the Archaeocidaridae and Palaeozoic Miocidaridae (Echinodermata, Echinoidea) and the origin of crown group echinoids

The archaeocidarids comprise the most derived stem group echinoids and have long been regarded as closely related to the crown group. The fossil record of echinoids in the Palaeozoic is, however, poor, so details surrounding the initial divergence of crown group echinoids are not well constrained. In order to better understand the phylogenetic relationships of the most derived stem group and most basal crown group echinoids, a phylogenetic analysis was undertaken of the Archaeocidaridae, including the genera Nortonechinus, Devonocidaris, Lepidocidaris, Polytaxicidaris and Archaeocidaris and the Palaeozoic miocidarid cidaroids from the genus Eotiaris. We found that Archaeocidaris appears to be paraphyletic with respect to crown group echinoids. Furthermore, we mapped character evolution along our phylogeny and found that the diversification of archaeocidarids and miocidarids may be linked to large-scale macroecological changes taking place in the late Palaeozoic, including increasing predation pressure and echinoid encrustation by epibionts. We compared the stratigraphical distribution of archaeocidarid and miocidarid occurrences to our resulting phylogenies, and found that the fit of our cladograms to the stratigraphic record of archaeocidarid occurrences is worse than other echinoid groups, supporting the idea that the imbricate plated archaeocidarids have a poor fossil record. In the course of carrying out these analyses, we also felt it necessary to describe a new species of Archaeocidaris, Archaeocidaris ivanovi sp. nov. We also provide novel descriptions and interpretations for Devonocidaris primaevus, Archaeocidaris brownwoodensis, Archaeocidaris apheles and revise the synonymy of Archaeocidaris legrandensis and ?Eotiaris meurevillensis, which may be a crown group echinoid