A RT/PCR-partial restriction enzymatic mapping (PREM) method for the molecular characterisation of the large satellite RNAs of Arabis mosaic virus isolates.

The satellite RNA of the grapevine isolate NW of Arabis mosaic virus (ArMV) was cloned and sequenced, and showed 75% identity at the nucleotide level to the satellite RNA of the lilac isolate of ArMV. In order to survey ArMV isolates from various geographical origins and natural hosts for the presence of large satellite RNAs and analyse their degree of variability, a RT/PCR-partial restriction enzymatic mapping (PREM) method was developed. The method is based on the incorporation of 5-methyl-dCTP in the RT/PCR reaction, and the subsequent digestion of the RT/PCR products by methyl-sensitive restriction enzymes. Satellites RNAs were detected by RT/PCR in eight isolates out of 47, six of them originating from grapevine, one from hop and one from lilac. The partial restriction digestion patterns allowed to distinguish six different types of satellites. Cloning and sequencing of the different satellites confirmed these results, the PREM proving able to discriminate sequences with 96% identity. The sizes of the different satellites varied between 1092 and 1139 nucleotides, their encoded proteins between 338 and 360 amino acids. Conserved domains were found in the amino and carboxy-termini between the sequences of the proteins encoded by the satellites of the different isolates of ArMV.

Preclinical and clinical studies with latex from Ficus glabrata HBK, a traditional intestinal anthelminthic in the Amazonian area.

Ficus glabrata latex has been a well-known anthelminthic remedy in the neotropical regions since ancient times. The latex has been commercially exploited for decades because of its content of the proteolytic enzyme-complex ficin. A safe dosage regimen with direct use of the latex has been elucidated to control intestinal helminthiasis in the Indian and non-Indian rural population. Helminthiasis was common in three Amazonian villages, field bases for the clinical study, with an overall prevalence of 92%. Specific prevalences were: Ascaris 68%, Strongyloides 42%, Trichuris 41%, Ancylostoma/Necator 26% and Taenia 1%. Variation in the biological activity of the latex was estimated by using a milk coagulating test. Pharmacological studies with live Ascaris demonstrated a lethal effect at concentrations down to 0.05% latex in physiological saline solution. A clinical trial on 181 persons has resulted in a recommended dosage of 1.0 cm3 of prepared latex/kg per day for 3 days to be repeated every 3 months.