Molecular Target Validation of Aspartate Transcarbamoylase from Plasmodium falciparum by Torin 2.

Malaria is a tropical disease that kills about half a million people around the world annually. Enzymatic reactions within pyrimidine biosynthesis have been proven to be essential for Plasmodium proliferation. Here we report on the essentiality of the second enzymatic step of the pyrimidine biosynthesis pathway, catalyzed by aspartate transcarbamoylase (ATC). Crystallization experiments using a double mutant ofPlasmodium falciparum ATC (PfATC) revealed the importance of the mutated residues for enzyme catalysis. Subsequently, this mutant was employed in protein interference assays (PIAs), which resulted in inhibition of parasite proliferation when parasites transfected with the double mutant were cultivated in medium lacking an excess of nutrients, including aspartate. Addition of 5 or 10 mg/L of aspartate to the minimal medium restored the parasites' normal growth rate. In vitro and whole-cell assays in the presence of the compound Torin 2 showed inhibition of specific activity and parasite growth, respectively. In silico analyses revealed the potential binding mode of Torin 2 to PfATC. Furthermore, a transgenic ATC-overexpressing cell line exhibited a 10-fold increased tolerance to Torin 2 compared with control cultures. Taken together, our results confirm the antimalarial activity of Torin 2, suggesting PfATC as a target of this drug and a promising target for the development of novel antimalarials.

Human T-lymphotropic virus type I (HTLV-I) seroprevalence in Jamaica. I. Demographic determinants.

During 1985 and 1986, the authors measured antibodies to human T-lymphotropic virus type I (HTLV-I) in a cohort of 13,260 Jamaicans from all parts of the island who applied for food-handling licenses. HTLV-I seroprevalence was strongly age and sex dependent, rising from 1.7% (10-19 years) to 9.1% (greater than or equal to 70 years) in men and from 1.9% (10-19 years) to 17.4% (greater than or equal to 70 years) in women. In a logistic regression analysis, women were more likely to be seropositive than were men, and farmers, laborers, and the unemployed were more likely to be HTLV-I seropositive than were those reporting student or professional occupations. In men, African ethnicity was associated with HTLV-I seropositivity in the univariate analysis but was not a risk factor after adjustment for age and sex. There was a trend toward higher age-stratified HTLV-I seroprevalence among younger women who reported more pregnancies, but older multigravidas had lower rates of HTLV-I seropositivity. Persons born outside Jamaica had significantly lower seroprevalence than did those born in Jamaica, but they were of slightly different ethnic and occupational compositions than those born in Jamaica.

Human T lymphotropic virus type I antibody patterns: evidence of difference by age and risk group.

Detection of human T lymphotropic virus type I (HTLV-I) antibody was assessed on 368 sera from subjects with different clinical features and from different parts of the world. Enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay for purified p24 antibodies (p24-RIA) used as screening tests agreed in 88.7% of the sera. The results from 247 selected sera were compared with western blot (WB). WB was reactive in sera five to 25 times more dilute than the last positive ELISA or p24-RIA, but different WB batches varied in sensitivity. ELISA was more sensitive than p24-RIA, and p24-RIA was more specific than ELISA. Indeterminate WB interpretations were common (25.5%). Most seropositive intravenous drug abusers had unusually strong p24 bands by WB. Among healthy individuals, positive WB reactivity increased with age, whereas indeterminate reactivity declined (P = .034). Thus, more-sensitive and -specific HTLV-I antibody tests are needed.