Akiko Tsubouchi

Toxoplasma gondii infection: Relationship between seroprevalence and risk factors among primary schoolchildren in the capital areas of Democratic Republic of São Tomé and Príncipe, West Africa

BackgroundThe status of Toxoplasma gondii infection among primary schoolchildren (PSC) of the Democratic Republic of São Tomé and Príncipe (DRSTP), West Africa, remains unknown to date.MethodsA serologic survey and risk factors associated T. gondii infection among PSC in the DRSTP was assessed by the latex agglutination (LA) test and a questionnaire interview including parents’ occupation, various uncomfortable symptoms, histories of eating raw or undercooked food, drinking unboiled water, and raising pets, was conducted in October 2010. Schoolchildren from 4 primary schools located in the capital areas were selected, in total 255 serum samples were obtained by venipuncture, of which 123 serum samples were obtained from boys (9.8 ± 1.4 yrs) and 132 serum samples were obtained from girls (9.7 ± 1.3 yrs).ResultsThe overall seroprevalence of T. gondii infection was 63.1% (161/255). No significant gender difference in seroprevalence was found between boys (62.6%, 77/123) and girls (63.6%, 84/132) (p = 0.9). The older age group of 10 years had insignificantly higher seroprevalence (69.9%, 58/83) than that of the younger age group of 8 year olds (67.7%, 21/31) (p = 0.8). It was noteworthy that the majority of seropositive PSC (75.8%, 122/161) had high LA titers of ≥1: 1024, indirectly indicating acute or repeated Toxoplasma infection. Parents whose jobs were non-skilled workers (73.1%) showed significantly higher seroprevalence than that of semiskilled- (53.9%) or skilled workers (48.8%) (p < 0.05). Children who had a history of raising cats also showed significantly higher seroprevalence than those who did not (p < 0.001).Children who claimed to have had recent ocular manifestation or headache, i.e. within 1 month, seemed to have insignificantly higher seroprevalence than those who did not (p > 0.05).ConclusionsParents’ educational level and cats kept indoors seemed to be the high risk factors for PSC in acquisition of T. gondii infection. While, ocular manifestation and/or headache of PSC should be checked for the possibility of being T. gondii elicited. Measures such as improving environmental hygiene and intensive educational intervention to both PSC and their parents should be performed immediately so as to reduce T. gondii infection of DRSTP inhabitants including PSC and adults.

Critical importance of the de novo pyrimidine biosynthesis pathway for Trypanosoma cruzi growth in the mammalian host cell cytoplasm.

The intracellular parasitic protist Trypanosoma cruzi is the causative agent of Chagas disease in Latin America. In general, pyrimidine nucleotides are supplied by both de novo biosynthesis and salvage pathways. While epimastigotes-an insect form-possess both activities, amastigotes-an intracellular replicating form of T. cruzi-are unable to mediate the uptake of pyrimidine. However, the requirement of de novo pyrimidine biosynthesis for parasite growth and survival has not yet been elucidated. Carbamoyl-phosphate synthetase II (CPSII) is the first and rate-limiting enzyme of the de novo biosynthetic pathway, and increased CPSII activity is associated with the rapid proliferation of tumor cells. In the present study, we showed that disruption of the T. cruzi cpsII gene significantly reduced parasite growth. In particular, the growth of amastigotes lacking the cpsII gene was severely suppressed. Thus, the de novo pyrimidine pathway is important for proliferation of T. cruzi in the host cell cytoplasm and represents a promising target for chemotherapy against Chagas disease.

Differential remodelling of peroxisome function underpins the environmental and metabolic adaptability of diplonemids and kinetoplastids.

The remodelling of organelle function is increasingly appreciated as a central driver of eukaryotic biodiversity and evolution. Kinetoplastids including Trypanosoma and Leishmania have evolved specialized peroxisomes, called glycosomes. Glycosomes uniquely contain a glycolytic pathway as well as other enzymes, which underpin the physiological flexibility of these major human pathogens. The sister group of kinetoplastids are the diplonemids, which are among the most abundant eukaryotes in marine plankton. Here we demonstrate the compartmentalization of gluconeogenesis, or glycolysis in reverse, in the peroxisomes of the free-living marine diplonemid, Diplonema papillatum. Our results suggest that peroxisome modification was already under way in the common ancestor of kinetoplastids and diplonemids, and raise the possibility that the central importance of gluconeogenesis to carbon metabolism in the heterotrophic free-living ancestor may have been an important selective driver. Our data indicate that peroxisome modification is not confined to the kinetoplastid lineage, but has also been a factor in the success of their free-living euglenozoan relatives.

Molecular interaction of the first 3 enzymes of the de novo pyrimidine biosynthetic pathway of Trypanosoma cruzi

The first 3 reaction steps of the de novo pyrimidine biosynthetic pathway are catalyzed by carbamoyl-phosphate synthetase II (CPSII), aspartate transcarbamoylase (ATC), and dihydroorotase (DHO), respectively. In eukaryotes, these enzymes are structurally classified into 2 types: (1) a CPSII-DHO-ATC fusion enzyme (CAD) found in animals, fungi, and amoebozoa, and (2) stand-alone enzymes found in plants and the protist groups. In the present study, we demonstrate direct intermolecular interactions between CPSII, ATC, and DHO of the parasitic protist Trypanosoma cruzi, which is the causative agent of Chagas disease. The 3 enzymes were expressed in a bacterial expression system and their interactions were examined. Immunoprecipitation using an antibody specific for each enzyme coupled with Western blotting-based detection using antibodies for the counterpart enzymes showed co-precipitation of all 3 enzymes. From an evolutionary viewpoint, the formation of a functional tri-enzyme complex may have preceded-and led to-gene fusion to produce the CAD protein. This is the first report to demonstrate the structural basis of these 3 enzymes as a model of CAD. Moreover, in conjunction with the essentiality of de novo pyrimidine biosynthesis in the parasite, our findings provide a rationale for new strategies for developing drugs for Chagas disease, which target the intermolecular interactions of these 3 enzymes.

Prevalence of intestinal parasitic infections among primary schoolchildren in areas devoid of sanitation in northwestern Kingdom of Swaziland, Southern Africa

Intestinal parasitic infections (IPIs) including helminths and protozoa are estimated to affect around 3·5 billion people globally and 450 million are ill as a result of these infections, the major...

Seroprevalence, seroconversion, and risk factors for toxoplasmosis among pregnant women in Taipei, Taiwan

Herein, we determined the seroprevalence, seroconversion, and risk factors associated with Toxoplasma gondii (T. gondii) infection among pregnant women in Taipei, Taiwan. Pregnant women attending antenatal consultation in a Taipei medical center were invited, and 104 women completed a self-administered structured questionnaire. Venous blood samples were collected during the first and third trimester after consent was obtained. Serum IgG and IgM antibodies (Abs) as well as IgG avidity were analyzed using an enzyme-linked fluorescent assay. Of the samples collected in the first trimester, seven were seropositive for IgG Abs and one was seropositive for IgG + IgM Abs with a borderline avidity index, resulting in an overall seroprevalence of 7.7%. No statistically significant association was found between toxoplasmosis and age, pregnancy history, or any risk factors. Seroconversion was not detected from paired sera between the first and third trimesters. Pregnant women with senior high school education level or those who claimed to knowing Toxoplasma exhibited a significantly higher seroprevalence than those with bachelor degree (P = 0.05) or those who claimed not to have this knowledge (P = 0.05). Therefore, failure to understand the importance of T. gondii infection and the prevention measures resulted in the development of toxoplasmosis among these women.