Full coding hepatitis E virus genotype 3 genome amplification method.

Hepatitis E virus (HEV) genotype 3 produces zoonotic infection associated with the consumption of infected animals. HEV infections can become chronic in immunocompromised (IC) patients. The viral genome has three well defined open reading frames (ORF1, ORF2 and ORF3) within which various domains and functions have been described. This paper (i) describes a new method of complete sequencing of the HEV coding region through overlapping PCR systems, (ii) establishes a consensus sequence and polymorphic positions (PP) for each domain, and (iii) analyzes the complete coding sequence of an IC patient. With regard to the consensus, a high percentage of PP was observed in protease (PP=19%) and the X domain (PP=22%) within ORF1, the N-terminal region of the S domain (PP=22%) in ORF2, and the P1 (PP=35%) and P2 (PP=25%) domains in ORF3. In contrast, the ORF1 Y, ORF2 S, ORF2 M and ORF3 D1 domains were conserved in the reference sequences (0.40, 1, 0.70 and 0% of PP, respectively). The sequence from the IC patient had more mutations in the RpRp (D1235G, Q1242R, S1454T, V1480I, I1502 V, K1511R, G1373 V, E1442D, V1693 M), the terminal ORF2 S- domain (F10L, S26T, G36S, S70P, A105 V, I113 V), the X domain (T938 M, T856 V, S898A) and the helicase (S1014N, S975T, Q1133 K).

Detección del gnnoma del virus de la hepatitis (vhe) en muestras de heces de cerdos en plantas de beneficio de antioquia, Colombia / Detection o hepaatitis e vius (hev) genome from pigs feces samples in slauughterhouses in antioquia, Colombia

El Virus de la Hepatitis E (VHE) es uno de los agentes causales de enfermedad hepática aguda en humanos, aunque también puede inducir hepatitis crónica en pacientes inmunocomprometidos. Existen cuatro genotipos que generan enfermedad en humanos: los genotipos 1 y 2 asociados con brotes epidémicos por consumo de aguas contaminadas y los genotipos 3 y 4 de trasmisión zoonótica, implicados en brotes esporádicos en países desarrollados donde el cerdo es el principal reservorio. En Colombia existe evidencia serológica de la infección en humanos y cerdos: se ha detectado el genoma viral en hígados de cerdos en plantas de beneficio y expendios de carne; sin embargo no se conoce lo suficiente sobre la infección en el país. Con el fin de determinar si los cerdos del departamento de Antioquia (Colombia) están excretando VHE en la edad del beneficio, se obtuvieron 152 muestras de heces de cerdos en cinco plantas de beneficio de distintas regiones del departamento en las que se determinó la presencia del genoma viral por medio RT-PCR. El porcentaje de positividad hallado fue del 26.9% (41/152); se encontró, además, que los cerdos que provenían de las subregiones Norte y Oriente de Antioquia tuvieron el menor (11.6%) y mayor (58.3%) porcentaje de muestras positivas, respectivamente. Estos resultados indican que los cerdos en el momento de sacrificio están excretando el virus a través de sus heces y que el VHE está circulando en las diferentes subregiones del departamento.

The Hepatitis E Virus (HEV) is one of the causative agents of acute liver disease in humans, although it can also lead to chronic hepatitis in immunocompromised patients. There are four genotypes that generate human disease: genotypes 1 and 2, associated with outbreaks due to consumption of contaminated waters, and genotypes 3 and 4 by zoonotic transmission, implicated in sporadic outbreaks in developed countries, where pigs are the main reservoir. In Colombia there is serological evidence of infection in humans and pigs: the viral genome has been detected in livers of pigs at slaughterhouses and butcher shops; however is not enough known about the infection in the country. In order to find out whether pigs in Antioquia (Colombia) are excreting the virus, the presence of the viral genome by RT-PCR was determined in 152 samples of pig feces obtained at five slaughterhouses of Antioquia, which came from different regions of the department. The percentage of positivity was 26.9% (41/152) and pigs that came from the North and East subregions of Antioquia had the lowest (11.6%) and higher (58.3%) percentage of positive samples, respectively. These results indicate that pigs at slaughter age are excreting the virus in their feces and that HEV is circulating in different subregions of the department.

Cytotoxic and antiviral activities of Colombian medicinal plant extracts of the Euphorbia genus.

Forty-seven plant extracts of 10 species of the genus Euphorbia (Euphorbiaceae) used by Colombian traditional healers for the treatment of ulcers, cancers, tumors, warts, and other diseases, were tested in vitro for their potential antitumour (antiproliferative and cytotoxic) and antiherpetic activity. To evaluate the capacity of the extracts to inhibit the lytic activity of herpes simplex virus type 2 (HSV-2) and the reduction of viability of infected or uninfected cell cultures, the end-point titration technique (EPTT) and the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colorimetric assay were used, respectively. The therapeutic index of the positive extracts for the antiviral activity was determined by calculating the ratio CC50 (50% cytotoxic concentration) over IC50 (50% inhibitory concentration of the viral effect). Five of the 47 extracts (11%) representing 3 out of 10 Euphorbia species (30%) exhibited antiherpetic action; the highest activity was found in the leaf/stem water-methanol extracts from E. cotinifolia and E. tirucalli. The therapeutic indexes of these two plant species were > 7.1; these extracts exhibited no cytotoxicity. Six extracts (13%) representing 4 plant species (40%) showed cytotoxic activity. The highest cytotoxicity was found in the dichloromethane extract obtained from E. cotinifolia leaves and the CC50 values for the most susceptible cell lines, HEp-2 and CHO, were 35.1 and 18.1 microgram/ml, respectively.

Cytotoxic and Antiviral Activities of Colombian Medicinal Plant Extracts of the Euphorbia genus

Forty-seven plant extracts of 10 species of the genus Euphorbia (Euphorbiaceae) used by Colombian traditional healers for the treatment of ulcers, cancers, tumors, warts, and other diseases, were tested in vitro for their potential antitumour (antiproliferative and cytotoxic) and antiherpetic activity. To evaluate the capacity of the extracts to inhibit the lytic activity of herpes simplex virus type 2 (HSV-2) and the reduction of viability of infected or uninfected cell cultures, the end-point titration technique (EPTT) and the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colorimetric assay were used, respectively. The therapeutic index of the positive extracts for the antiviral activity was determined by calculating the ratio CC50 (50 percent cytotoxic concentration) over IC50 (50 percent inhibitory concentration of the viral effect). Five of the 47 extracts (11 percent) representing 3 out of 10 Euphorbia species (30 percent) exhibited antiherpetic action; the highest activity was found in the leaf/stem water-methanol extracts from E. cotinifolia and E. tirucalli. The therapeutic indexes of these two plant species were > 7.1; these extracts exhibited no cytotoxicity. Six extracts (13 percent) representing 4 plant species (40 percent) showed cytotoxic activity. The highest cytotoxicity was found in the dichloromethane extract obtained from E. cotinifolia leaves and the CC50 values for the most susceptible cell lines, HEp-2 and CHO, were 35.1 and 18.1 æg/ml, respectively