Fasciola hepatica secretes cathepsin L proteases that facilitate the penetration of the parasite through the tissues of its host, and also participate in functions such as feeding and immune evasion. The major proteases, cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) are members of a lineage that gave rise to the human cathepsin Ls, Ks and Ss, but while they exhibit similarities in their substrate specificities to these enzymes they differ in having a wider pH range for activity and an enhanced stability at neutral pH. There are presently 13 Fasciola cathepsin L cDNAs deposited in the public databases representing a gene family of at least seven distinct members, although the temporal and spatial expression of each of these members in the developmental stage of F. hepatica remains unclear. Immunolocalisation and in situ hybridisation studies, using antibody and DNA probes, respectively, show that the vast majority of cathepsin L gene expression is carried out in the epithelial cells lining the parasite gut. Within these cells the enzyme is packaged into secretory vesicles that release their contents into the gut lumen for the purpose of degrading ingested host tissue and blood. Liver flukes also express a novel multi-domain cystatin that may be involved in the regulation of cathepsin L activity. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 have shown that these enzymes can induce protection, ranging from 33 to 79%, to experimental challenge with metacercariae of F. hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission. In this article we review the vaccine trials carried out over the past 8 years, the role of antibody and T cell responses in mediating protection and discuss the prospects of the cathepsin Ls in the development of first generation recombinant liver fluke vaccines. q

The phenomenon of asexual multiplication is rare in the animal kingdom, but it occurs in all main flatworm taxa. In the present paper, we review data regarding the presence of different forms of asexual multiplication in flatworms and argue that the presence of a population of totipotent or pluripotent stem cells, "neoblasts", is a primitive feature of decisive importance for the developing potential of flatworms. Next we present information on the role of stem cells in fission, head regeneration, and pharynx regeneration of planarians. Furthermore, the tracing of neoblasts in lower flatworms and cestodes is presented, and the results indicating heterogeneity of the neoblast pool are discussed. Finally, the mode by which the neoblasts are stimulated to divide, migrate, and differentiate and the nature of the interactions are discussed. We focus on (i) biogenic amines and neuropeptides, (ii) the role of neuropeptides in the early stage of regeneration, (iii) the evidence for the influences of growth factors and nitric oxide, and (iv) the influence of weak electromagnetic fields. We discuss the pattern in which a gradient system of morphogens and (or) a hierarchical system of inductions is expressed in development.

The physiological effects of selected classical transmitters and FMRFamide-related peptides (FaRPs) on dispersed muscle fibres from the marine turbellarian, Procerodes littoralis have been examined. Confocal scanning laser microscopy coupled with fluorescein isothiocyanate (FITC) or tetramethylrhodamine (TRITC)-labelled phalloidin revealed a highly developed body wall muscle system with circular, longitudinal and diagonal layers of muscle fibres. Dispersed muscle fibres contracted when depolarized by exposure to extracellular media with elevated K+ (15-100 mM) in a concentration-dependent manner, with a maximal response of 87% achieved at> or = 75 mM. 5-Hydroxytryptamine (5-HT) induced concentration-dependent muscle contraction between 0.01 and 1000 microM, with 10 microM producing a near maximal contraction response of 75%. Acetylcholine (ACh) had less pronounced excitatory effects (0.01-1000 microM), inducing contraction of only 32% of the fibres at 100 microM. The flatworm FMRFamide-related peptides (FaRPs), GYIRFamide, YIRFamide and GNFFRFamide each had concentration-dependent myocontractile effects, indicating the occurrence of at least 1 FaRP receptor on P. littoralis muscle fibres. At 10 microM peptide, GNFFRFamide induced contractions in < 40% of the muscle fibres examined, whereas YIRFamide and GYIRFamide induced contraction in 70 and 75% of muscle fibres, respectively. The order of potency of the peptides was: GYIRFamide > YIRFamide > GNFFRFamide. Pre-incubation of the muscle fibres in 5 microM 5-HT significantly reduced the responses to GYIRFamide, YIRFamide and 5-HT, while the responses to high K+ remained unaltered. Muscle fibres pre-incubated in GYIRFamide (0.1 microM) were also less responsive to 5-HT but not to ACh and high-K+. The GYIRFamide analogue, GYIRDFamide, did not induce muscle contraction (0.01-100 microM) per se, but when co-applied with the myoactive peptides GYIRFamide, YIRFamide or GNFFRFamide, it significantly blocked their ability to elicit contractions. This suggests that the peptides tested may act via a common muscle-based neuropeptide receptor. GYIRDFamide did not alter the contractile effects of high K+, 5-HT or ACh. Collectively, these results indicate that FaRPs, 5-HT and ACh all have the potential to cause muscle contraction in flatworms and that 5-HT and FaRPs alter muscle sensitivity to each other, but do not influence the ability of flatworm muscle fibres to contract.

No abstract

No abstract