Adult Schistosoma mansoni express cathepsin L proteinase activity.

This report presents the deduced amino acid sequence of a novel cathepsin L proteinase from Schistosoma mansoni, and describes cathepsin L-like activity in extracts of adult schistosomes. Using consensus primers specific for cysteine proteinases, gene fragments were amplified from adult S. mansoni cDNA by PCR and cloned.

The phylogeny of Neospora caninum.

Morphological studies by electron microscopy on the protozoan Neospora caninum have shown that this organism possesses a subcellular structure typical of parasites classified in the family Sarcocystidae, subclass Coccidiasina of the phylum Apicomplexa. Using a strategy based on DNA sequence analysis of products derived by asymmetric PCR to determine the nucleotide sequences, we have tested the validity of this classification by comparing the small subunit ribosomal RNA (18S rRNA) gene sequences of N. caninum with those of other parasitic protozoa classified in the phylum Apicomplexa.

Drug resistance to schistosomicides and other anthelmintics of medical significance.

It is usual for people to be infected for some period in life with parasitic worms, which may cause morbidity or even kill. Anthelmintics are used for the treatment and control of the human helminthiases, since no vaccines are yet available. Despite the widespread use of these compounds, drug resistance has become apparent only with antischistosomal chemotherapy, in contrast to the situation with other anti-infective agents in human medicine and with veterinary anthelmintics, where resistance is widespread.

Indications, evaluation, complications, and results of functional endoscopic sinus surgery in 200 patients.

Functional endoscopic sinus surgery (FESS) is a new and exciting treatment for chronic sinus disease. Our knowledge of the surgery continues to expand. A retrospective and prospective review of 200 patients undergoing FESS was undertaken at the Houston Ear, Nose, and Throat Clinic.

Differentiation of Toxoplasma gondii from closely related coccidia by riboprint analysis and a surface antigen gene polymerase chain reaction.

The tachyzoite of the human pathogen Toxoplasma gondii is morphologically indistinguishable from the proliferative stages of some other zoonotic coccidia, including Sarcocystis. To determine the identity of such coccidia obtained from human tissues and other sources, we compared riboprints (through restriction enzyme analysis of the polymerase chain reaction [PCR]-amplified small subunit rRNA gene) of the following protozoa: the RH and ts-4 strains of T. gondii, lines OH3 and S11, which are two recently isolated T.

Schistosoma mansoni: use of a cloned ribosomal RNA gene probe to detect restriction fragment length polymorphisms in the intermediate host Biomphalaria glabrata.

Adult susceptibility of Biomphalaria glabrata to Schistosoma mansoni infection is controlled by simple Mendelian genetics. In this study a molecular approach was used to determine the degree of genetic variation between well-defined lines of B. glabrata which are either resistant (10-R2) or susceptible (M-line) to S.

Characterization of a programmed alteration in an 18S ribosomal gene that accompanies the experimental induction of drug resistance in Schistosoma mansoni.

Stable resistance to the anthelmintic hycanthone can be produced in the human blood fluke Schistosoma mansoni by exposing immature parasites in mice to the drug. Within a single generation, genomic rearrangements, detected as rRNA-encoding DNA restriction fragment length polymorphisms (RFLPs), accompany the appearance of resistance in this model. One of these RFLPs, an approximately 3.

Cloning and characterization of a species-specific repetitive DNA sequence from Loa loa.

A genomic DNA library of Loa loa was constructed in lambda gt11 using EcoRI-digested DNA from microfilariae isolated from two West African patients. Screening with labeled L. loa DNA yielded several potential repetitive DNA clones.

Genomic variability in field populations of Schistosoma mansoni in Brazil as detected with a ribosomal gene probe.

A cloned fragment of the ribosomal gene of Schistosoma mansoni, pSM 389, which contains part of the small rRNA gene plus a portion of the nontranscribed intergenic spacer, was used in Southern hybridization analyses to investigate genomic variation in natural populations of S. mansoni in Brazil. Genomic DNAs were isolated from schistosomes from infected patients (some of whom did not respond to antischistosomal chemotherapy), and from snails from disparate geographic locations in Brazil.

Strongyloides stercoralis: identification of a protease that facilitates penetration of skin by the infective larvae.

Host invasion and tissue migration of several helminths have been linked to the expression and release of parasite-derived proteases. One of the most remarkable examples of tissue migration is that of larvae of the nematode parasite Strongyloides stercoralis, which can move through tissue at speeds of up to 10 cm per hour. We have shown the Strongyloides L3 larvae secrete a potent histolytic metalloprotease to facilitate their rapid migration.

A genomic change associated with the development of resistance to hycanthone in Schistosoma mansoni.

Ribosomal gene probes were used to investigate the genetic basis of drug resistance in schistosomes in a model where resistance to the anthelmintic hycanthone (HC) is generated by exposing immature worms to the drug. Two strains of Schistosoma mansoni, JHU and NMRI, were used. Drug resistance could be produced in the JHU strain by treatment with HC, but was also found to occur spontaneously.

Role of host antibody in the chemotherapeutic action of praziquantel against Schistosoma mansoni: identification of target antigens.

We have demonstrated previously in a mouse model that effective chemotherapy against Schistosoma mansoni with praziquantel (PZQ) is dependent upon an intact host antibody response. In the same study, it was found that worms recovered from PZQ-treated animals display surface-bound antibodies. In order to identify the target antigens of the antibodies involved in the synergy between PZQ and the immune response, monoclonal antibodies (mAbs) and polyclonal antisera recognizing different tegumental components were tested by indirect immunofluorescence (IF) assay for their ability to bind in vitro to the surface of 6-week-old schistosomes perfused from nude (athymic) mice 1 h after PZQ treatment.

Nematospiroides dubius: passive transfer of protective immunity to mice with monoclonal antibodies.

Nine hybridoma cell lines secreting monoclonal antibodies specific for Nematospiroides dubius were produced by fusion of the mouse myeloma cell line NS-1 to either spleen cells or mesenteric lymph node cells from mice repeatedly infected with N. dubius. Seven of the antibodies were identified as IgM and two as IgG1.

Antigens from the surface and excretions/secretions of the filariform larva of Strongyloides stercoralis.

The surface and excretory/secretory (ES) antigens of the infective, filariform larva (L3) of Strongyloides stercoralis were identified. These studies provide a basis for the purification of these proteins as diagnostic allergens for human strongyloidiasis. The Mr values of the surface and ES molecules were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, fluorography, or silver staining following the recovery of these molecules after the radiolabelling of living parasites.

The chemotherapeutic effect of praziquantel against Schistosoma mansoni is dependent on host antibody response.

To assess the role of host humoral immune responses in the mechanism of action of praziquantel (PZQ) against Schistosoma mansoni, the efficacy of the drug was compared in infected B cell-depleted (mu-suppressed) vs immunologically intact C3H/HeN mice. We found that PZQ was on the average only 20% as effective in eliminating adult schistosomes from mu-suppressed as compared with control animals. Indeed, in three of four experiments performed, the drug failed to significantly reduce adult worm burdens in the mu-suppressed mice.

Anti-schistosomal drugs: observations on the mechanism of drug resistance to hycanthone, and on the involvement of host antibodies in the mode of action of praziquantel.

This paper reports recent observations from our laboratory dealing with the anti-schistosome drugs hycanthone (HC) and praziquantel (PZQ). In particular, we discuss a laboratory model of drug resistance to HC in Schistosoma mansoni and show that drug sensitive and resistant lines of the parasite can be differentiated on the basis of restriction fragment length polymorphisms using homologous ribosomal gene probes. In addition, we summarize data demonstrating that effective chemotherapy of S.

Nematospiroides dubius: multiple infections in mice bred for immune responsiveness.

Allogeneic mice were reared over 5 generations in a 2-way selection for high (H) and low (L) immune responsiveness to the intestinal trichostrongylid nematode Nematospiroides dubius. After 5 generations of selective breeding, the H mice passed fewer N. dubius eggs and harbored fewer, smaller and less fecund worms than did the L mice.

Inheritance of immunity in mice to challenge infection with Nematospiroides dubius.

Two lines of mice (Mus musculus) were selectively reared over 10 generations for high (H) and low (L) levels of immune response to Nematospiroides dubius, an enteric nematode parasite. Filial and backcross families were derived from the two parent lines. The mode of inheritance of the trait, immune response to challenge infection with N.

Nematospiroides dubius: direct and correlated responses to selection for high and low immune responsiveness in mice.

High and low immune responder lines of mice were bred selectively from an allogeneic stock over 10 generations, based on their fecal parasite egg count assayed 3 weeks after reinfection with 100 Nematospiroides dubius larvae. By generation 10, (F10), the low immune response mice voided about 10 times as many fecal N. dubius eggs as the high immune response mice.

Susceptibility of rats to infection with Toxocara pteropodis.

Infective eggs of Toxocara pteropodis were administered to Wistar rats via oral and parenteral routes. Third-stage larvae were recovered from the livers of suckling young 8 days after oral infection, and from livers and lungs after intraperitoneal or subcutaneous inoculation of eggs. These larvae were short-lived as none were found in suckling mice killed 2 weeks post-infection.

Vaccination against Nematospiroides dubius in mice using adult worm extracts as antigens.

Various preparations of somatic Nematospiroides dubius antigens were emulsified in Freund's complete adjuvant and assayed in Quackenbush mice for efficacy as vaccines against homologous infection. Soluble and particulate fractions were given by two different routes. Only minimal protection was induced by vaccination with either soluble or particulate parasite fractions used independently when compared with the adjuvant control.