A catecholamine transporter from the human parasite Schistosoma mansoni with low affinity for psychostimulants

The trematode Schistosoma mansoni is the primary cause of schistosomiasis, a devastating neglected tropical disease that affects 200 million individuals. Identifying novel therapeutic targets for the treatment of schistosomiasis is therefore of great public interest. The catecholamines norepinephrine (NE) and dopamine (DA) are essential for the survival of the parasite as they cause muscular relaxation and a lengthening in the parasite and thereby control movement. Here we characterize a novel dopamine/norepinephrine transporter (SmDAT) gene transcript, from S. mansoni. The SmDAT is expressed in the adult form and in the sporocyst form (infected snails) of the parasite, and also in the egg and miracidium stage. It is absent in the cercariae stage but curiously a transcript missing the exon encoding transmembrane domain 8 was identified in this stage. Heterologous expression of the cDNA in mammalian cells resulted in saturable, dopamine transport activity with an apparent affinity for dopamine comparable to that of the human dopamine transporter. Efflux experiments reveal notably higher substrate selectivity compared with its mammalian counterparts as amphetamine is a much less potent efflux elicitor against SmDAT compared to the human DAT. Pharmacological characterization of the SmDAT revealed that most human DAT inhibitors including psychostimulants such as cocaine were significantly less potent in inhibiting SmDAT. Like DATs from other simpler organisms the pharmacology for SmDAT was more similar to the human norepinephrine transporter. We were not able to identify other dopamine transporting carriers within the completed parasite genome and we hypothesize that the SmDAT is the only catecholamine transporter in the parasite and could be responsible for not only clearing DA but also NE. (C) 2011 Elsevier B.V. All rights reserved.

Two allelic isoforms of the serotonin transporter from Schistosoma mansoni display electrogenic transport and high selectivity for serotonin

The human blood fluke Schistosoma mansoni is the primary cause of schistosomiasis, a debilitating disease that affects 200 million individuals in over 70 countries. The biogenic amine serotonin is essential for the survival of the parasite and serotonergic proteins are potential novel drug targets for treating schistosomiasis. Here we characterize two novel serotonin transporter gene transcripts, SmSERT-A and SmSERT-B, from S. mansoni. Southern blot analysis shows that the two mRNAs are the products of different alleles of a single SmSERT gene locus. The two SmSERT forms differ in three amino acid positions near the N-terminus of the protein. Both SmSERTs are expressed in the adult form and in the sporocyst form (infected snails) of the parasite, but are absent from all other stages of the parasite`s complex life cycle. Heterologous expression of the two cDNAs in mammalian cells resulted in saturable, sodium-dependent serotonin transport activity with an apparent affinity for serotonin comparable to that of the human serotonin transporter. Although the two SmSERTs are pharmacologically indistinguishable from each other, efflux experiments reveal notably higher substrate selectivity for serotonin compared with their mammalian counterparts. Several well-established substrates for human SERT including (+/-)MDMA, S-(+)amphetamine, RU 24969, and m-CPP are not transported by SmSERTs, underscoring the higher selectivity of the schistosomal isoforms. Voltage-clamp recordings of SmSERT substrate-elicited currents confirm the substrate selectivity observed in efflux experiments and suggest that it may be possible to exploit the electrogenic nature of SmSERT to screen for compounds that target the parasite in vivo. (C) 2009 Elsevier B.V. All rights reserved.

Dam worms

Human and animal infection rates with the Oriental schistosome have steadily declined in China over the last half-century, but the Three Gorges Dam may reverse this decline by creating new, or enlarging existing, ideal environments for the worm and its aquatic snail intermediate host.

Alpha-conotoxins: Nicotinic acetylcholine receptor antagonists as pharmacological tools and potential drug leads

Alpha-Conotoxins are small disulfide rich peptides from the venoms of marine cone snails. They target specific nicotinic acetylcholine receptor (nAChR) subtypes with high affinity and potency and are therefore valuable as neurophamacological probes and potential drug leads. This article gives a general overview of the chemical and biological features of alpha -conotoxins, including their pharmacology, binding interactions and structure. A detailed analysis of recently reported three-dimensional structures from members of different subfamilies of the alpha -conotoxins, including those with 3/5, 4/3, 4/6 and 4.7 spacings of their two intracysteine loops is given. The structures are generally well defined and represent useful frameworks for the display of amino acid residues to target molecules.

Biogeographical concordance and efficiency of taxon indicators for establishing conservation priority in a tropical rainforest biota

Prioritizing areas for conservation requires the use of surrogates for assessing overall patterns of biodiversity. Effective surrogates will reflect general biogeographical patterns and the evolutionary processes that have given rise to these and their efficiency is likely to lie influenced by several factors, including the spatial scale of species turnover and the overall congruence of the biogeographical history. We examine patterns of surrogacy for insects, snails, one family of plants and vertebrates from rainforests of northeast Queensland, an area characterized by high endemicity and an underlying history of climate-induced vicariance. Nearly all taxa provided some level of prediction of the conservation values For others. However, despite an overall correlation of the patterns of species richness and complementarity, the efficiency of surrogacy was highly asymmetric.. snails and insects were strong predictors of conservation priorities for vertebrates, but not vice versa. These results confirm predictions that taxon surrogates can be effective in highly diverse tropical systems where there is a strong history of vicariant biogeography, but also indicate that correlated patterns for species richness and/or complementarity do not guarantee that one taxon will be efficient as a surrogate for another. In our case, the highly diverse and narrowly distributed invertebrates were more efficient as predictors than the less diverse and more broadly distributed vertebrates.

Two new classes of conopeptides inhibit the alpha 1-adrenoceptor and noradrenaline transporter

Cone snails use venom containing a cocktail of peptides ('conopeptides') to capture their prey. Many of these peptides also target mammalian receptors, often with exquisite selectivity. Here we report the discovery of two new classes of conopeptides. One class targets alpha (1)-adrenoceptors (rho -TIA from the fish-hunting Conus tulipa), and the second class targets the neuronal noradrenaline transporter (chi -MrIA and chi -MrIB from the mollusk-hunting C. marmoreus). rho -TIA and chi -MrIA selectively modulate these important membrane-bound proteins. Both peptides act as reversible non-competitive inhibitors and provide alternative avenues for the identification of inhibitor drugs.

The spread of Angiostrongylus cantonensis in Australia

Until the recent establishment of Angiostrongylus cantonensis in North America. Australia was the only developed region endemic for this parasite. Almost 50 years ago the life cycle was elucidated there, in the city of Brisbane, and the first human infections probably occurred in 1959. From the 1970s, increasing numbers of autochthonous infections have been reported along the central east coast of the continent (southeast Queensland and northern New South Wales), involving humans, rats, dogs, horses, flying foxes and marsupials. Ten years ago, the parasite was discovered in Sydney, almost 1,000 km to the south, in dogs. In that city, it has since been diagnosed as a cause of neurological disease in increasing numbers of dogs, flying foxes, marsupials and zoo primates. Presumably, these infections resulted from the ingestion of snails or slugs, and it seems that virtually all species of native and exotic terrestrial molluscs can serve as intermediate hosts. It is not known how the parasite was introduced to this continent, or how it has spread over such an extensive territory, although eventually its range could encompass the entire east coast, and potentially other regions. It is also not known if the almost identical, native species, A. mackerrasae, is able to infect people (or other non-rodent hosts). All worms recovered to date, from one fatal human case, and from many animal infections, have been confirmed as A. cantonensis.

Observations on the miraxonal attraction exercised by sexually immature or adult Biomphalaria glabrata infected or not by Schistosoma mansoni

OBJECTIVE: The influence of age and the presence of secondary sporocysts in the miraxonal attraction exercised by Biomphalaria glabrata on miracidia of Schistosoma mansoni of the BH strain were studied. MATERIAL AND METHOD: A glass apparatus containing two compartments joined by a tube and previously tested in other experiments, was used. Specimens of B. glabrata or its snail conditioned water (SCW) selected before the first oviposition (sexually immature), after the first oviposition (adult), with or without secondary sporocysts, were used to attract the miracidia. RESULTS: It was noted that snails or their SCW containing secondary sporocysts lost the ability to attract miracidia. The sexual maturity of the snail did not influence miraxonal attraction.

Experimental infection with Schistosoma mansoni of Biomphalaria straminea from different parts of the Northeast of Brazil

In the western part of the State of Bahia Biomphalaria straminea and B. glabrata both occur, but in the majority of cases they do not share the same habitat. In the State of Ceará, however, B. straminea is the sole snail host of Schistosoma mansoni. In this survey, no naturally infected B. straminea was found among snails collected from Bahia and Ceará, evidently because of the very low infection rates. The susceptibility of laboratory-reared specimens to infection with a Puerto Rican strain of S. mansoni was then tested experimentally. In general, the snails showed very low susceptibility. The infection rates were 1.1% among snails from Redenção (Ceará); 2.3% in those from Pentecoste (Ceará); 2.9% in snails from São Desidério (Bahia), while they were very high among an albino strain (NIH) of B. glabrata used as control. Another group of B. straminea from São Desidério was exposed to a Bahian strain of S. mansoni and the infection rate was still very low (3.6%) Apparently, the very low susceptibility of B. straminea, despite high snail density, is correlated with moderate infection rates with S. mansoni among humans, as shown by the results of stool examinations conducted by SUCAM in the municipalities of Redenção and Pentecoste, in Ceará.

Periphyton used as food for Biomphalaria tenagophila (Gastropoda, Planorbidae)

Snails reared in cages colonized by periphyton grew from 5.0mm to 8.8mm shell diameter, each laid 0.5 batch of eggs per day and the overall survival during the period was 75%

Strain variation in the infectivity of Schistosoma mansoni for Biomphalaria glabrata

Five strains of Schistosoma mansoni resistant and susceptible to schistosomicides were studied for infectivity of 2 strains of Biomphalaria glabrata one of Puerto Rican origin and the other of Brazilian origin. Puerto Rican strains of S. Mansoni developed more slowly and had a lower infectivity in Brazilian B. glabrata than did the Brazilian S. mansoni. However, Brazilian S. Mansoni developed as well in Puerto Rican snails as in Brazilian snails, indicating that drug resistant strains could easily be moved by travel of infected persons from one area to another.

Schistosomiasis mansoni in the municipality of Pedro de Toledo (São Paulo, Brazil) where the Biomphalaria tenagophila is the snail host: I - Prevalence in human population (

Due to the scarce information about the epidemiological features of schistosomiasis in which the vector is Biomphalaria tenagophila, an investigation was carried in Pedro de Toledo in 1980 where such peculiarity is observed. Stool examinations (Kato-Katz method) were performed in 4,741 individuals (22.8% positive to Schistosoma mansoni eggs) of this 583 had previously received chemoterapy and 4,158 remainders, untreated. The schistosomiasis prevalence in those two groups where respectively 31.7% and 21.6%. Epidemiological investigation showed that 83.6% were autochthonous cases from the studied area: the autochthonous prevalence rate, and the intensity of infection in the untreated autochthonous cases were higher in males than in females; the intensity in the latter untreated group was low, 58.5 eggs/g feces (geometric mean). Moreover, according to the age groups the intensity of infections correlated well (r s = 0.745) with the prevalence rates. Schistosomiasis was verified to occur mostly during the leisure time and by the use of water streams for housework in rural zone. Only 0.4% out of 1,137 snails was positive for S. mansoni cercariae, apparently unchanged from the 1978 study when the human prevalence was 12.0%. The studied area presented differences and similarities in relation to the other Brazilian areas were the main intermediate host is B. glabrata.

First register of Biomphalaria straminea Dunker, 1848, in Santa Catarina State

The first register of Biomphalaria straminea, vector species of schistosomiasis, in Santa Catarina State, South Brazil is described. The specimens were obtained from two ornamental aquaria of private residences. In both cases the ornamental plants and/or fishes were bought from the same supplier. The presence of this species was later confirmed in the farm where these plants and fishes are cultivated, in the city of Governador Celso Ramos, also in Santa Catarina State. The occurrence in natural environments was later detected in two different places of the Island of Santa Catarina.

Evaluation of temporal, seasonal and geographic stability of the molluscicidal property of Euphorbia splendens latex

Laboratory tests with aqueous solutions of Euphorbia splendens var. hislopii latex have demonstrated seasonal stability of the molluscicidal principle, with LD90 values of 1.14 ppm (spring), 1.02 ppm (fall), 1.09 ppm (winter), and 1.07 ppm (summer) that have been determined against Biomphalaria tenagophila in the field. Assays on latex collected in Belo Horizonte and Recife yielded LD90 values similar to those obtained with the reference substance collected in Rio de Janeiro (Ilha do Governador), demonstrating geographic stability of the molluscicidal effect. The molluscicidal action of aqueous dilutions of the latex in natura, centrifuged (precipitate) and lyophilized, was stable for up to 124 days at room temperature (in natura) and for up to 736 days in a common refrigerator at 10 to 12ºC (lyophilized product). A 5.0 ppm solution is 100% lethal for snails up to 13 days after preparation, the effect being gradually lost to almost total inactivity by the 30th day. This observation indicated that the active principle is instable. These properties together with the wide distribution of the plant, its resistance and adaptation to the tropical climate, its easy cultivation and the easy obtention of latex and preparation of the molluscicidal solution, make this a promising material for large-scale use in the control of schistosomiasis

Biological and morphological characteristics of Schistosoma mansoni from Ribeira Valley, State of São Paulo, Brazil: I - susceptibility of Biomphalaria tenagophila snail to sympatric S. mansoni strain

In the São Paulo State, Brazil, where the Biomphalaria tenagophila is the intermediate host, the Ribeira Valley is an important endemic schistosomiasis mansoni area. During last eleven years there has been intense control measures focusing on schistosomiasis. The efforts have been concentrated in the municipalities of Pedro de Toledo and Itariri. We determined the susceptibility of B. tenagophila to sympatric strain of S. mansoni, both recently isolated from Itariri field. In 1988, this strain was isolated and maintained in the experimental model: Swiss mice - sympatric B. tenagophila. The second generation of the worm was evaluated. The snail were divided in the three groups of 60 snails each. One group was exposed to 1 miracidium and other to 10. The third group was the control. The mortality and the shedding of cercariae were checked during 78 days. After that, the positive snails were observed until they ceased to shed cercariae. The exposed molluscs showed mortality rates of 23% and 31% and infection indexes were of 8% and 60% to 1 and 10 miracidia respectively. The mortality was of 22% in the control group. The periods of shedding cercariae in the two groups were 82 and 104 days. We can conclude that B. tenagophila is an effective intermediate host to the sympatric strain of S. mansoni sympatric strain