Eeyarestatin I, an inhibitor of the valosin-containing protein, exhibits potent virucidal activity against the flaviviruses.

Cellular responses to stress generally lead to the activation of the endoplasmic reticulum-associated protein degradation (ERAD) pathway. Several lines of study support that ERAD may be playing a proviral role during flaviviral infection. A key host factor in ERAD is the valosin-containing protein (VCP), an ATPase which ushers ubiquitin-tagged proteins to degradation by the proteasome. VCP exhibits different proviral activities, such as engaging in the biogenesis of viral replication organelles and facilitating flavivirus genome uncoating after the viral particle entry. To investigate the possible antiviral value of drugs targeting VCP, we tested two inhibitors: eeyarestatin I (EEY) and xanthohumol (XAN). Both compounds were highly effective in suppressing Zika virus (ZIKV) and Usutu virus (USUV) replication during infection in cell culture. Further analysis revealed an unexpected virucidal activity for EEY, but not for XAN. Preincubation of ZIKV or USUV with EEY before inoculation to cells resulted in significant decreases in infectivity in a dose- and time-dependent manner. Viral genomes in samples previously treated with EEY were more sensitive to propidium monoazide, an intercalating agent, with 10- to 100-fold decreases observed in viral RNA levels, supporting that EEY affects viral particle integrity. Altogether, these results support that EEY is a strong virucide against two unrelated flaviviruses, encouraging further studies to investigate its potential use as a broad-acting drug or the development of improved derivatives in the treatment of flaviviral infection.

Detection of Mayaro virus in Aedes aegypti mosquitoes circulating in Goiânia-Goiás-Brazil.

The Arboviral diseases are caused by arthropod-borne viruses, such as Mayaro virus (MAYV), the etiological agent of Mayaro fever. This disease has been drawing the attention of the public health authorities for the increased number of cases likely due to virus adaptation for survival to urban areas as well as infection and multiplication in other vectors insects. Therefore, this work aimed to identify the MAYV infecting Aedes aegypti mosquitoes in Goiânia, the capital of state of Goiás, Brazil. For the development of study, the larvae of A. aegypti were collected in Basic Health Units from different regions of Goiânia then the larvae were grown to adult mosquitoes in controlled laboratory conditions. The female mosquitoes were submitted to the procedure of head and body separation. The RNAs obtained from these samples were analyzed by real-time PCR for identification of arboviruses. We only detect the presence of MAVY in the mosquitoes, in this sense our findings suggest that A. aegypti harbor MAYV in different anatomical sites, and potentially the process of vertical transmission of MAYV can occur in this vector.

Genotypic and phenotypic characterization of attaching and effacing Escherichia coli (AEEC) isolated from children with and without diarrhoea in Londrina, Brazil.

Attaching and effacing Escherichia coli (AEEC) have been implicated in diarrhoea in humans in several countries. A total of 919 E. coli strains, isolated from 125 children with diarrhoea and 98 without diarrhoea, was investigated by PCR for the presence of the EAF, bfp, eae and stx genes. Thirty-four of these isolates were found to carry the eae gene; they were isolated from 27 (79.4 %) children with diarrhoea and seven (20.6 %) controls, in the city of Londrina, Brazil. These strains were investigated for their genotypic and phenotypic characteristics. Different genetic profiles were observed; strains containing the eae gene alone were most common (47.1 %). The characteristic genetic profile of typical enteropathogenic E. coli (EPEC), eae, bfp and EAF, was only found in isolates from children with diarrhoea. The stx gene was not detected in any of the 34 strains studied. Ten (29.4 %) strains were negative in the fluorescent actin-staining test. Localized adhesion (LA) was the most common pattern of adhesion (44.1 %), followed by the aggregative adhesion (AA) (23.5 %) and localized adhesion-like (LAL) (14.7 %) patterns. The results showed a strong association between strains presenting the LA pattern and diarrhoea. Forty-seven per cent of the strains studied belonged to classical O-serogroups of EPEC. The most common serotype found was O119 : H6; these isolates all showed the LA pattern, were positive for fluorescent actin-staining and were associated with diarrhoea. Intimin beta was detected in seven strains, four of which belonged to serotype O119 : H6 and three to serotype ONT : H7; all were associated with diarrhoea. On the other hand, intimin epsilon was detected in two strains of serotype O111 : H38 and one of serotype ONT : H19, isolated from children without diarrhoea. To our knowledge, this is the first report of the occurrence of intimin epsilon in strains of E. coli isolated from humans in Brazil.