Molecular systematics of the Dichopetala genus group (Orthoptera: Phaneropteridae)

Introducción: El grupo de géneros Dichopetala se propuso recientemente después de una revisión del género Dichopetala Brunner von Wattenwyl, 1878. Actualmente, el grupo consta de ocho géneros y 44 especies distribuidas desde el sur de los Estados Unidos hasta el sur de México. Este acuerdo genérico, se basó únicamente en evidencia morfológica y se acompañó por discusiones sobre nuevos géneros erigidos, para los cuales no se probó su monofilia. Objetivo: Evaluar las relaciones filogenéticas entre especies representativas de los ocho géneros del grupo Dichopetala. Métodos: Generamos secuencias de ADN para un gen mitocondrial (Citocromo oxidasa I: COI) y dos marcadores de genes nucleares (28S, Histona III: H3), e incluimos especies de otros géneros de Phaneropterinae para probar la monofilia del grupo en estudio. Utilizamos modelos evolutivos bayesianos y de máxima verosimilitud. Resultados: Se respalda la monofilia del grupo Dichopetala y la monofilia de los géneros Dichopetala, Obolopteryx, Planipollex, Mactruchus y Rhabdocerca. Además, los géneros Acanthorintes y Pterodichopetala como parafiléticos. Los marcadores mitocondriales también sugieren que los géneros Rhabdocerca y Acanthorintes ampliamente distribuidos, pueden en realidad contener varias especies no vistas previamente. Conclusión: Se proporciona la primera contribución a la filogenia del grupo de Dichopetala y una definición filogenética y morfológica más robusta de algunos de los géneros involucrados.

Introduction: The Dichopetala genus group was proposed recently after revision of the genus Dichopetala Brunner von Wattenwyl, 1878. Currently, the group consists of eight genera and 44 species distributed from Southern United States to Southern Mexico. This generic arrangement was based solely on morphological evidence, and was accompanied by discussions on new erected genera, for which their monophyly was not tested. Objective: To assess the phylogenetic relationships among representative species of the eight genera of the Dichopetala group. Methods: We generated DNA sequences for one mitochondrial (Cytochrome oxidase I: COI) and two nuclear (28S, Histone III: H3) gene markers, and included species of other Phaneropterinae genera to test the monophyly of the ingroup; Bayesian and maximum likelihood evolutionary models were used. Results: The monophyly of the Dichopetala group and the monophyly of genera Dichopetala, Obolopteryx, Planipollex, Mactruchus and Rhabdocerca is supported. In addition, Acanthorintes and Pterodichopetala were recovered as paraphyletic. The mitochondrial markers also suggest that the widely distributed genera Rhabdocerca and Acanthorintes may actually contain various overlooked species. Conclusions: The first contribution on the Phylogeny of the Dichopetala group, and a more robust phylogenetic and morphological definition of some of the genera involved are provided.

Cellular and molecular studies of the effects of a selective COX-2 inhibitor celecoxib in the cardiac cell line H9c2 and their correlation with death mechanisms

Cardiovascular disease is one of the leading causes of death worldwide, and evidence indicates a correlation between the inflammatory process and cardiac dysfunction. Selective inhibitors of cyclooxygenase-2 (COX-2) enzyme are not recommended for long-term use because of potentially severe side effects to the heart. Considering this and the frequent prescribing of commercial celecoxib, the present study analyzed cellular and molecular effects of 1 and 10 µM celecoxib in a cell culture model. After a 24-h incubation, celecoxib reduced cell viability in a dose-dependent manner as also demonstrated in MTT assays. Furthermore, reverse transcription-polymerase chain reaction analysis showed that the drug modulated the expression level of genes related to death pathways, and Western blot analyses demonstrated a modulatory effect of the drug on COX-2 protein levels in cardiac cells. In addition, the results demonstrated a downregulation of prostaglandin E2 production by the cardiac cells incubated with celecoxib, in a dose-specific manner. These results are consistent with the decrease in cell viability and the presence of necrotic processes shown by Fourier transform infrared analysis, suggesting a direct correlation of prostanoids in cellular homeostasis and survival.

Isolation in HRT-18 cells and molecular analysis of a BCoV strain from diarrheic feces of naturally infected calves

Bovine coronavirus (BCoV) may cause acute diarrhea in newborn calves, leading to significant economic losses for cattle farmers. There are several diagnostic techniques used to detect BCoV in calf fecal samples, but virus isolation still has advantages for antigenic and genomic characterization. This study describes the isolation in HRT-18 cells and molecular characterization of Brazilian BCoV wild-type strains. Three fecal samples from diarrheic 30 day-old calves were inoculated in HRT-18 cell monolayers, which were then evaluated for HA titers and tested using semi-nested PCR followed by RFLP and sequencing. Two samples were successfully isolated and presented HA titers of 16 and 32 units per 25 mL. The results were confirmed using semi-nested PCR and RFLP. Molecular analyses identified a cell culture-adapted strain and a wild-type strain that were genetically similar (99 percent) to each other, but more distinct than BCoV strains circulating in other countries, even in the conserved N gene.

O coronavírus bovino (BCoV) pode causar diarreia aguda em bezerros recém-nascidos, ocasionando consideráveis perdas econômicas para a pecuária bovina. Várias técnicas de diagnóstico podem ser empregadas na detecção do BCoV a partir de amostras fecais de bezerros. Porém, o isolamento do BCoV em cultivo celular apresenta a vantagem de possibilitar a caracterização antigênica e molecular da estirpe viral. O presente estudo descreve o isolamento em células HRT-18, e a caracterização molecular de estirpes brasileiras do BCoV. Três amostras de fezes diarreicas de bezerros com 30 dias de idade foram inoculadas em culturas de células HRT-18. Os isolados foram avaliados por hemaglutinação (HA) e por uma semi-nested PCR seguida de RFLP e sequenciamento. Duas amostras foram isoladas e a confirmação foi verificada na semi-nested PCR e também RFLP. Na HA os títulos foram de 16 e 32 unidades por 25 mL. Análises moleculares identificam a estirpe adaptada em cultura celular e uma estirpe selvagem, como estirpes de BCoV semelhantes (99 por cento) entre si, mas distintas das circulantes em outros países, mesmo em um gene de uma proteína conservada (gene N).