Identification of enzymes that have helminth-specific active sites and are required for Rhodoquinone-dependent metabolism as targets for new anthelmintics.

Soil transmitted helminths (STHs) are major human pathogens that infect over a billion people. Resistance to current anthelmintics is rising and new drugs are needed. Here we combine multiple approaches to find druggable targets in the anaerobic metabolic pathways STHs need to survive in their mammalian host. These require rhodoquinone (RQ), an electron carrier used by STHs and not their hosts. We identified 25 genes predicted to act in RQ-dependent metabolism including sensing hypoxia and RQ synthesis and found 9 are required. Since all 9 have mammalian orthologues, we used comparative genomics and structural modeling to identify those with active sites that differ between host and parasite. Together, we found 4 genes that are required for RQ-dependent metabolism and have different active sites. Finding these high confidence targets can open up in silico screens to identify species selective inhibitors of these enzymes as new anthelmintics.

A New Species of Gordionus (Nematomorpha: Gordiida) from the Rocky Mountains of New Mexico.

The 19 genera within the phylum Nematomorpha contain approximately 350 species. The cosmopolitan genus Gordionus Müller, 1926 contains about 58 species, 6 of which occur in the contiguous United States of America. Recently, 2 new Gordionus species were described from high-altitude streams within the southern Rocky Mountains, near Santa Fe, New Mexico. Here we describe another new Gordionus species, from a high-altitude stream in the southern Rocky Mountains, from near Taos, New Mexico. The sites consisted of temporary puddles and a small human-made stream at 3,175-3,250-m altitude in aspen/pine woodland. Gordionus lokeri n. sp. has 1 areole type, which varies in shape and size between and within body regions. Midbody areoles are elongated, polygonal, or triangular, shingled, with the raised side of the areole serrated. The interareolar space is narrow, containing few bristles. The male cloacal opening is surrounded inside and outside by narrow branching bristles that bifurcate or trifurcate deeply; the furcae then subdivide several times terminally. Adhesive warts are lacking. Genetic data, consisting of partial cytochrome c oxidase subunit I (COI) gene sequences, clearly separated G. lokeri n. sp. from other Nearctic species. This is the third Gordionus species described from high-elevation streams in the Rocky Mountains. It appears that this high-altitude habitat represents the preferential niche for numerous species of this genus, and thus future work should focus on describing gordiid diversity in other parts of the Rocky Mountains.

A review on the druggability of a thiol-based enzymatic antioxidant thioredoxin reductase for treating filariasis and other parasitic infections.

Understanding and elucidating the mechanism of host-pathogen interactions are the major area of interest among the Parasitologists all around the globe. Starting from the origin on mother earth parasites have searched for successful strategies to invade their respective host for the sake of survivability and eventually succeeded to manage in the unfriendly environment inside the host's body. Parasite-generated antioxidants are potent enough to combat the oxidative challenges inside the host body and within its own as well. Antioxidant enzymes are tremendously important as they are directly related to the survival of the parasites. The thiol-based antioxidant enzymes (glutathione reductase and thioredoxin reductase) have dragged much attention of the researchers to date. In this regard, among the thiol-based antioxidants, particularly the Thioredoxin reductase (TrxR), is known to be present in a number of parasitic organisms have pulled the researchers. Therefore, selective targeting of TrxR can emerge as a novel capital for developing suitable adulticidal candidate for treating filariasis and other helminth infections. This review tries to assemble the existing knowledge of the parasitic TrxR and how these can be utilized as a druggable target in cases of filariasis and other helminth infections has been discussed.

Cysteine proteases as digestive enzymes in parasitic helminths.

We briefly review cysteine proteases (orthologs of mammalian cathepsins B, L, F, and C) that are expressed in flatworm and nematode parasites. Emphasis is placed on enzyme activities that have been functionally characterized, are associated with the parasite gut, and putatively contribute to degrading host proteins to absorbable nutrients [1-4]. Often, gut proteases are expressed as multigene families, as is the case with Fasciola [5] and Haemonchus [6], presumably expanding the range of substrates that can be degraded, not least during parasite migration through host tissues [5]. The application of the free-living planarian and Caenorhabditis elegans as investigative models for parasite cysteine proteases is discussed. Finally, because of their central nutritive contribution, targeting the component gut proteases with small-molecule chemical inhibitors and understanding their utility as vaccine candidates are active areas of research [7].

Cysteine proteases during larval migration and development of helminths in their final host.

Neglected tropical diseases caused by metazoan parasites are major public health concerns, and therefore, new methods for their control and elimination are needed. Research over the last 25 years has revealed the vital contribution of cysteine proteases to invasion of and migration by (larval) helminth parasites through host tissues, in addition to their roles in embryogenesis, molting, egg hatching, and yolk degradation. Their central function to maintaining parasite survival in the host has made them prime intervention targets for novel drugs and vaccines. This review focuses on those helminth cysteine proteases that have been functionally characterized during the varied early stages of development in the human host and embryogenesis.

Advances in kinome research of parasitic worms - implications for fundamental research and applied biotechnological outcomes.

Protein kinases are enzymes that play essential roles in the regulation of many cellular processes. Despite expansions in the fields of genomics, transcriptomics and bioinformatics, there is limited information on the kinase complements (kinomes) of most eukaryotic organisms, including parasitic worms that cause serious diseases of humans and animals. The biological uniqueness of these worms and the draft status of their genomes pose challenges for the identification and classification of protein kinases using established tools. In this article, we provide an account of kinase biology, the roles of kinases in diseases and their importance as drug targets, and drug discovery efforts in key socioeconomically important parasitic worms. In this context, we summarise methods and resources commonly used for the curation, identification, classification and functional annotation of protein kinase sequences from draft genomes; review recent advances made in the characterisation of the worm kinomes; and discuss the implications of these advances for investigating kinase signalling and developing small-molecule inhibitors as new anti-parasitic drugs.

TRP channels in schistosomes.

Praziquantel (PZQ) is effectively the only drug currently available for treatment and control of schistosomiasis, a disease affecting hundreds of millions of people worldwide. Many anthelmintics, likely including PZQ, target ion channels, membrane protein complexes essential for normal functioning of the neuromusculature and other tissues. Despite this fact, only a few classes of parasitic helminth ion channels have been assessed for their pharmacological properties or for their roles in parasite physiology. One such overlooked group of ion channels is the transient receptor potential (TRP) channel superfamily. TRP channels share a common core structure, but are widely diverse in their activation mechanisms and ion selectivity. They are critical to transducing sensory signals, responding to a wide range of external stimuli. They are also involved in other functions, such as regulating intracellular calcium and organellar ion homeostasis and trafficking. Here, we review current literature on parasitic helminth TRP channels, focusing on those in schistosomes. We discuss the likely roles of these channels in sensory and locomotor activity, including the possible significance of a class of TRP channels (TRPV) that is absent in schistosomes. We also focus on evidence indicating that at least one schistosome TRP channel (SmTRPA) has atypical, TRPV1-like pharmacological sensitivities that could potentially be exploited for future therapeutic targeting.

The Role of Xenobiotic-Metabolizing Enzymes in Anthelmintic Deactivation and Resistance in Helminths.

Xenobiotic-metabolizing enzymes (XMEs) modulate the biological activity and behavior of many drugs, including anthelmintics. The effects of anthelmintics can often be abolished by XMEs when the drugs are metabolized to an inefficient compound. XMEs therefore play a significant role in anthelmintic efficacy. Moreover, differences in XMEs between helminths are reflected by differences in anthelmintic metabolism between target species. Taking advantage of the newly sequenced genomes of many helminth species, progress in this field has been remarkable. The present review collects up to date information regarding the most important XMEs (phase I and phase II biotransformation enzymes; efflux transporters) in helminths. The participation of these XMEs in anthelmintic metabolism and their possible roles in drug resistance are evaluated.

Metabolic Enzymes of Helminth Parasites: Potential as Drug Targets.

Metabolic pathways that extract energy from carbon compounds are essential for an organism's survival. Therefore, inhibition of enzymes in these pathways represents a potential therapeutic strategy to combat parasitic infections. However, the high degree of similarity between host and parasite enzymes makes this strategy potentially difficult. Nevertheless, several existing drugs to treat infections by parasitic helminths (worms) target metabolic enzymes. These include the trivalent antimonials that target phosphofructokinase and Clorsulon that targets phosphoglycerate mutase and phosphoglycerate kinase. Glycolytic enzymes from a variety of helminths have been characterised biochemically, and some inhibitors identified. To date none of these inhibitors have been developed into therapies. Many of these enzymes are externalised from the parasite and so are also of interest in the development of potential vaccines. Less work has been done on tricarboxylic acid cycle enzymes and oxidative phosphorylation complexes. Again, while some inhibitors have been identified none have been developed into drug-like molecules. Barriers to the development of novel drugs targeting metabolic enzymes include the lack of experimentally determined structures of helminth enzymes, lack of direct proof that the enzymes are vital in the parasites and lack of cell culture systems for many helminth species. Nevertheless, the success of Clorsulon (which discriminates between highly similar host and parasite enzymes) should inspire us to consider making serious efforts to discover novel anthelminthics, which target metabolic enzymes.

Synthetic and natural protease inhibitors provide insights into parasite development, virulence and pathogenesis.

Protease function is essential to many biological systems and processes. In parasites, proteases are essential for host tissue degradation, immune evasion, and nutrition acquisition. Helminths (worms) depend on several classes of proteases for development, host tissue invasion and migration, and for degradation of host hemoglobin and serum proteins. The protozoa, which cause malaria, depend on both cysteine and aspartic proteases to initiate host hemoglobin digestion. Other types of proteases are involved in erythrocyte cell invasion and cell exit. Surface metalloproteases in kinetoplastids are implicated in the evasion of complement-mediated cell lysis and cell entry. Cysteine proteases in Entamoeba facilitate invasion of the host colon. Giardia utilizes a cysteine protease for both encystation and excystation. This review will summarize published data using protease inhibitors as tools to identify the function of parasite proteases in the development, virulence, and pathogenesis of parasites; as well as the role of endogenous parasite protease inhibitors in regulation.

[Research progress on cathepsin F of parasitic helminths].

Cathepsin F is an important member of papain-like subfamily in cysteine protease family. Cathepsin F of helminth parasites can hydrolyze the specific substrate, degrade host protein such as hemoglobin for nutrition, and be involved in invasion into host tissue. Therefore, cathepsin F serves as a potential target for parasitic disease immunodiagnosis, vaccine design and anti-parasite drug screening. This article reviews the structural characteristics and mechanisms of cathepsin F, and research advances on cathepsin F of parasitic helminths.

Molecular mimicry between cockroach and helminth glutathione S-transferases promotes cross-reactivity and cross-sensitization.

BACKGROUND: The extensive similarities between helminth proteins and allergens are thought to contribute to helminth-driven allergic sensitization. OBJECTIVE: The objective of this study was to investigate the cross-reactivity between a major glutathione-S transferase allergen of cockroach (Bla g 5) and the glutathione-S transferase of Wuchereria bancrofti (WbGST), a major lymphatic filarial pathogen of humans. METHODS: We compared the molecular and structural similarities between Bla g 5 and WbGST by in silico analysis and by linear epitope mapping. The levels of IgE, IgG, and IgG(4) antibodies were measured in filarial-infected and filarial-uninfected patients. Mice were infected with Heligmosomoides bakeri, and their skin was tested for cross-reactive allergic responses. RESULTS: These 2 proteins are 30% identical at the amino acid level with remarkable similarity in the N-terminal region and overall structural conservation based on predicted 3-dimensional models. Filarial infection was associated with IgE, IgG, and IgG(4) anti-Bla g 5 antibody production, with a significant correlation between antibodies (irrespective of isotype) to Bla g 5 and WbGST (P< .0003). Preincubation of sera from cockroach-allergic subjects with WbGST partially depleted (by 50%-70%) anti-Bla g 5 IgE, IgG, and IgG(4) antibodies. IgE epitope mapping of Bla g 5 revealed that 2 linear N-terminal epitopes are highly conserved in WbGST corresponding to Bla g 5 peptides partially involved in the inhibition of WbGST binding. Finally, mice infected with H bakeri developed anti-HbGST IgE and showed immediate-type skin test reactivity to Bla g 5. CONCLUSION: These data demonstrate that helminth glutathione-S transferase and the aeroallergen Bla g 5 share epitopes that can induce allergic cross-sensitization.

Blunting the knife: development of vaccines targeting digestive proteases of blood-feeding helminth parasites.

Proteases are pivotal to parasitism, mediating biological processes crucial to worm survival including larval migration through tissue, immune evasion/modulation and nutrient acquisition by the adult parasite. In haematophagous parasites, many of these proteolytic enzymes are secreted from the intestine (nematodes) or gastrodermis (trematodes) where they act to degrade host haemoglobin and serum proteins as part of the feeding process. These proteases are exposed to components of the immune system of the host when the worms ingest blood, and therefore present targets for the development of anti-helminth vaccines. The protective effects of current vaccine antigens against nematodes that infect humans (hookworm) and livestock (barber's pole worm) are based on haemoglobin-degrading intestinal proteases and act largely as a result of the neutralisation of these proteases by antibodies that are ingested with the blood-meal. In this review, we survey the current status of helminth proteases that show promise as vaccines and describe their vital contribution to a parasitic existence.

The global cysteine peptidase landscape in parasites.

The accumulation of sequenced genomes has expanded the already sizeable population of cysteine peptidases from parasites. Characterization of a few of these enzymes has ascribed key roles to peptidases in parasite life cycles and has also shed light on mechanisms of pathogenesis. Here we discuss recent observations on the physiological activities of cysteine peptidases of parasitic organisms, paired with a global view of all cysteine peptidases from the MEROPS database grouped by similarity. This snapshot of the landscape of parasite cysteine peptidases is complex and highly populated, suggesting that expansion of research beyond the few 'model' parasite peptidases is now timely.

Xenobiotic metabolizing enzymes and metabolism of anthelminthics in helminths.

Anthelminthics remain the only accessible means in the struggle against helminth parasites, which cause significant morbidity and mortality in man and farm animals. The treatment of helminthic infections has become problematic because of frequent drug resistance of helminth parasites. The development of drug resistance can be facilitated by the action of xenobiotic metabolizing enzymes (XMEs). In all organisms, XMEs serve as an efficient defense against the potential negative action of xenobiotics. The activities of XMEs determine both desired and undesired effects of drugs, and the knowledge of drug metabolism is necessary for safe, effective pharmacotherapy. While human and mammalian XMEs have been intensively studied for many years, XMEs of helminth parasites have undergone relatively little investigation, so far. However, many types of XMEs, including oxidases, reductases, hydrolases, transferases, and transporters, have been described in several helminth species. XMEs of helminth parasites may protect these organisms from the toxic effects of anthelminthics. In case of certain anthelminthics, metabolic deactivation was reported in helminth larvae and/or adults. Moreover, if a helminth is in the repeated contact with an anthelminthic, it defends itself against the chemical stress by the induction of biotransformation enzymes or transporters. This induction can represent an advantageous defense strategy of the parasites and may facilitate the drug-resistance development.

Enzymatic antioxidant systems in helminth parasites.

Parasitic helminths have a coexistence with mammalian hosts whereby they survive for several years in known hostile conditions of their hosts. Many explanations exist describing how these parasitic helminths are able to survive. In the last years, a lot of studies have focused on both enzymatic and non-enzymatic antioxidant systems now shown to exist in these parasites and which may serve as defence tactics against the host-generated oxygen radicals. The relevance of antioxidant enzymes is confirmed by the fact that some of these molecules represent putative protective anti-parasite vaccines (i.e. in schistosomiasis). This review tries to compile what is known to date of the enzymatic antioxidant systems in selected parasitic helminths.

Helminth pathogen cathepsin proteases: it's a family affair.

Helminth pathogens express papain-like cysteine peptidases, termed cathepsins, which have important roles in virulence, including host entry, tissue migration and the suppression of host immune responses. The liver fluke Fasciola hepatica, an emerging human pathogen, expresses the largest cathepsin L cysteine protease family yet described. Recent phylogenetic, biochemical and structural studies indicate that this family contains five separate clades, which exhibit overlapping but distinct substrate specificities created by a process of gene duplication followed by subtle residue divergence within the protease active site. The developmentally regulated expression of these proteases correlates with the passage of the parasite through host tissues and its encounters with different host macromolecules.

Flavonoids and related compounds in parasitic disease control.

Flavonoids are natural plant compounds increasingly used in therapeutic applications. Their large spectrum of activities depends on their structures and cellular targets. Most recent research shows they are promising drugs for controlling human and animal parasitic diseases. Their multiple effects make it difficult to understand their modes of action, but some of them have been elucidated. This review also deals with their toxicity in mammals.