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Molecular and Biochemical Parasitology

Clare Halliday, Karen Billington, Ziyin Wang, Samuel Dean, Jack Daniel Sunter, Richard John Wheeler
The kinetoplastids Trypanosoma brucei and Leishmania mexicana are eukaryotes with a highly structured cellular organisation that is reproduced with great fidelity in each generation. The pattern of signal from a fluorescently tagged protein can define the specific structure/organelle that this protein localises to, and can be extremely informative in phenotype analysis in experimental perturbations, life cycle tracking, post-genomic assays and functional analysis of organelles. Using the vast coverage of protein subcellular localisations provided by the TrypTag project, an ongoing project to determine the localisation of every protein encoded in the T...
December 11, 2018: Molecular and Biochemical Parasitology
Damien M O'Halloran
The pace of research towards a genetic model to understand the unique molecular biology of parasitic nematodes has increased recently. This research has developed a diverse suite of genetic tools for a variety of parasitic nematodes. CRISPR/Cas9 technology in particular offers much promise as a game changing tool for researchers studying parasitic nematodes. Unlike RNAi, which depends on diverse nematode effectors to silence gene expression, the effectors for CRISPR/Cas9 mutations are typically supplied by the experimenter, making gene editing via CRISPR/Cas9 ideal for testing on genetically intractable nematode systems...
December 7, 2018: Molecular and Biochemical Parasitology
Itoko Hayashi, Seiji Kanda, Pheophet Lamaningao, Nobuyuki Mishima, Toshimasa Nishiyama
This study focuses on the host-parasite relationship of human Ascaris lumbricoides, which is a parasite of the small intestine and is also one of the commonest parasites worldwide. As part of this investigation, we examined the host-parasite relationship assuming that there is a common antigenicity, shared protein between A. lumbricoides and human small intestinal mucosa, using molecular techniques. We obtained three DNA clones from human colon cDNA library by screening for anti-A. lumbricoides polyclonal antibodies...
November 24, 2018: Molecular and Biochemical Parasitology
Neyder Contreras Puentes, Erika Rodríguez Cavallo, Darío Méndez Cuadro
Deficiency of glucose-6-phosphate dehydrogenase (G6PD) and sickle cell trait (SCT) are described as the polymorphic disorders prevalent in erythrocytes. Both are considered the result of the selective pressure exerted by Plasmodium parasites over human genome, due to a certain degree of resistance to the clinical symptoms of severe malaria. There exist in both a prooxidant environment that favors the oxidative damage on membrane proteins, which probably is part of molecular protector mechanisms. Nevertheless, mechanisms are not completely understood at molecular level for each polymorphism yet, and even less if are commons for several of them...
November 22, 2018: Molecular and Biochemical Parasitology
Jingyi Liu, Zhirong Fu, Lars Hellman, Staffan G Svärd
Giardia intestinalis is a protozoan parasite and the causative agent of giardiasis, a common diarrheal disease. Cysteine protease (CP) activities have been suggested to be involved in Giardia's pathogenesis and we have recently identified and characterized three secreted Giardia CPs; CP14019, CP16160 and CP16779. Here we have studied the cleavage specificity of these CPs using substrate phage display and recombinant protein substrates. The phage display analyses showed that CP16160 has both chymase and tryptase activity and a broad substrate specificity...
November 17, 2018: Molecular and Biochemical Parasitology
Danielle Mn Moura, Osvaldo P de Melo Neto, Mark Carrington
A Trypanosoma brucei cell line is described that produces a visual readout of proteasome activity. The cell line contains an integrated transgene encoding an ubiquitin-green fluorescent protein (GFP) fusion polypeptide responsive to the addition of proteasome inhibitors. A modified version of T. brucei ubiquitin unable to be recognized by deubiquitinases (UbG76 V) was fused to eGFP and constitutively expressed. The fusion protein is unstable but addition of the proteasome inhibitor lactacystin stabilizes it and leads to visually detectable GFP...
November 13, 2018: Molecular and Biochemical Parasitology
Dietmar Steverding, Bogdan I Florea, Herman S Overkleeft
Previous studies indicated that the proteasome of the protozoan parasite Trypanosoma brucei is particularly sensitive to inhibition of the trypsin-like activity. In this study, three newly developed β2 subunit-specific inhibitor (LU-102, LU-002c and LU-002i) were tested for their ability to block the trypsin-like activity of the trypanosomal proteasome. At 10 μM, none of the compounds affected the proteasomal trypsin-like activity in cell lysates of bloodstream forms of T. brucei. On the other hand, leupeptin, a well-established β2 inhibitor, supressed the proteasomal trypsin-like activity within trypanosome cell lysates with a 50% inhibitory concentration of 2 μM demonstrating the inhibitability of the trypsin-like activity of the T...
November 13, 2018: Molecular and Biochemical Parasitology
Yosser Zina Abdelkrim, Emna Harigua-Souiai, Mourad Barhoumi, Josette Banroques, Arnaud Blondel, Ikram Guizani, N Kyle Tanner
The antifungal agent 6-aminocholestanol targets the production of ergosterol, which is the principle sterol in many fungi and protozoans; ergosterol serves many of the same roles as cholesterol in animals. We found that it also is an effective inhibitor of the translation-initiation factor eIF4AI from mouse (eIF4AIMus ) and the Trypanosomatid parasite Leishmania (LieIF4A). The eIF4A proteins belong to the DEAD-box family of RNA helicases, which are ATP-dependent RNA-binding proteins and RNA-dependent ATPases...
October 23, 2018: Molecular and Biochemical Parasitology
Ahmad F Eweas, Gamal Allam
Schistosomiasis represents a world health major problem affecting more than 206 million people worldwide. Up to date, praziquantel (PZQ) is the sole chemotherapeutic agent used in clinics for the treatment of schistosomiasis. The resistance to PZQ chemotherapy that has been emerged against some schistosome phenotypes represents the most serious PZQ-related problem so far. Therefore, it is clear that there is a substantial need to develop novel and effective antischistosomal agents in order to ensure the effective drug control of schistosomiasis in the future...
October 4, 2018: Molecular and Biochemical Parasitology
Drahomíra Faktorová, Anita Bär, Hassan Hashimi, Katherine McKenney, Aleš Horák, Achim Schnaufer, Mary Anne T Rubio, Juan D Alfonzo, Julius Lukeš
Ribosome biosynthesis, best studied in opisthokonts, is a highly complex process involving numerous protein and RNA factors. Yet, very little is known about the early stages of pre-18S rRNA processing even in these model organisms, let alone the conservation of this mechanism in other eukaryotes. Here we extend our knowledge of this process by identifying and characterizing the essential protein TbUTP10, a homolog of yeast U3 small nucleolar RNA-associated protein 10 - UTP10 (HEATR1 in human), in the excavate parasitic protist Trypanosoma brucei...
September 21, 2018: Molecular and Biochemical Parasitology
Jeong-Ho Kim, Jong-Oh Kim, Chan-Hyeok Jeon, U-Hwa Nam, Sathiyamoorthy Subramaniyam, Seung-Il Yoo, Jun-Hyung Park
We analyzed transcriptome profiles of Anisakis simplex (Nematoda: Anisakidae) 3rd (ASL3) and 4th larvae (ASL4) obtained by RNA-seq, to understand the molecular pathways linked to parasite survival and discover stage-enriched gene expressions. ASL3 were collected from chum salmon and ASL4 were obtained by in vitro culture. Whole transcriptome sequencing was conducted with Illumina sequencer, and de novo assembly was conducted. 47,179 and 41,934 genes were expressed in ASL3 and ASL4 transcriptomes. Of them, 17,633 were known and 29,546 were unmapped sequence for ASL3...
December 2018: Molecular and Biochemical Parasitology
Nikolay G Kolev, Trisha K Ramsdell, Christian Tschudi
Trypanosoma brucei relies on two types of variant surface glycoprotein (VSG) expression sites (ESs) for RNA polymerase I (Pol I) transcription of VSG pre-mRNA. Trypanosomes developing into infectious metacyclic cells in the tsetse vector use metacyclic VSG ESs (MESs) and proliferating parasites in the mammalian host deploy bloodstream VSG ESs (BESs). Unlike the monocistronic MESs, BESs are polycistronic and their highly conserved promoters differ considerably from the MES promoters. The significance of the divergent sequences of MES and BES promoters remains to be determined...
December 2018: Molecular and Biochemical Parasitology
Karla L R Batista, Carolina R Silva, Valdenice F Santos, Rafael C Silva, Renato R Roma, Ana L E Santos, Raquel O Pereira, Plínio Delatorre, Bruno A M Rocha, Alexandra M S Soares, Livio M Costa-Júnior, Claudener S Teixeira
Haemonchus contortus is one of the most economically important parasites infecting small ruminants worldwide. This nematode has shown a great ability to develop resistance to anthelmintic drugs, calling for the development of alternative control approaches. Because lectins recognize and bind to specific carbohydrates and glycan structures present in parasites, they can be considered as an alternative to develop new antiparasitic drugs. Accordingly, this work aimed to investigate the anthelmintic effect of Canavalia brasiliensis (ConBr) lectin against H...
September 11, 2018: Molecular and Biochemical Parasitology
Vinicius José Maschio, Veridiana Gomes Virginio, Henrique Bunselmeyer Ferreira, Marilise Brittes Rott
Acanthamoeba castellanii is a free-living organism widely distributed in the environment that may cause disease. This protozoan exists in two forms, an infective trophozoite and a dormant cyst. The trophozoites are able to cause keratitis and granulomatous amoebic encephalitis in humans. Keratitis is an acute, sight threatening infection of cornea with potential to cause permanent blindness without prompt treatment. However, the lack of suspicion and the low awareness about these amoebae besides of the absence of commercially available immunodiagnostic tests may delay an accurate diagnosis...
September 8, 2018: Molecular and Biochemical Parasitology
D Ditgen, E M Anandarajah, A Reinhardt, A E Younis, S Witt, J Hansmann, E Lorenz, M García-Hernández, D Paclik, H Soblik, A Jolodar, P H Seeberger, E Liebau, N W Brattig
Helminths are complex pathogens that ensure their long-term survival by influencing the immune responses of their host. Excretory/secretory products (ESP) can exert immunoregulatory effects which foster parasite survival. Galectins represent a widespread group of β-galactoside-binding proteins which are involved in a multitude of biological processes operative in parasite-host interaction. We had earlier identified seven galectins in Strongyloides ratti, four of them detected in the ESP of distinct developmental stages of the parasite...
September 1, 2018: Molecular and Biochemical Parasitology
María Victoria Solana, María Fernanda Domínguez, Silvana Scarcella, Santiago Radio, Pablo Smircich, Silvina Fernández, Hugo Solana, José F Tort
The use of Triclabendazole for controlling fasciolosis is compromised by increased drug resistance affecting livestock and humans. Although the mode of action of TCBZ is still unknown, putative candidates and markers of resistance have been advanced. A single nucleotide polymorphism (T687 G) in F. hepatica PGP was proposed as marker of resistance in a small scale study of European susceptible and resistant flukes, but the association was not found in Australian samples. The T687 G SNP was absent in more than 40 samples from 2 TCBZ-resistant and 3 susceptible isolates across Latin America here analyzed...
September 2018: Molecular and Biochemical Parasitology
Huafang Shi, Kiantra Butler, Christian Tschudi
We previously established an in vitro differentiation system based on the inducible expression of the RNA binding protein 6 (RBP6), which initiated differentiation of Trypanosoma brucei non-infectious procyclics to infectious metacyclics (MFs). However, further differentiation to bloodstream forms (BFs) required infection of mice. Here we report the serendipitous isolation of a single point mutation in RBP6 (Q109K), whose expression not only generated MFs, but purified MFs continued the developmental cycle in vitro to BFs expressing variant surface glycoprotein-2 (VSG-2), formerly known as VSG 221...
September 2018: Molecular and Biochemical Parasitology
Ahmad Syibli Othman, Jing-Wen Lin, Blandine M Franke-Fayard, Hans Kroeze, Fiona J A van Pul, Séverine Chevalley-Maurel, Jai Ramesar, Catherin Marin-Mogollon, Matthijs M Jore, Merribeth J Morin, Carole A Long, Robert Sauerwein, Ashley Birkett, Kazutoyo Miura, Chris J Janse, Shahid M Khan
The transmission-blocking vaccine candidate Pfs48/45 from the human malaria parasite Plasmodium falciparum is known to be difficult to express in heterologous systems, either as full-length protein or as correctly folded protein fragments that retain conformational epitopes. In this study we express full-length Pfs48/45 in the rodent parasite P. berghei. Pfs48/45 is expressed as a transgene under control of the strong P. berghei schizont-specific msp1 gene promoter (Pfs48/45@PbMSP1). Pfs48/45@PbMSP1 schizont-infected red blood cells produced full-length Pfs48/45 and the structural integrity of Pfs48/45 was confirmed using a panel of conformation-specific monoclonal antibodies that bind to different Pfs48/45 epitopes...
September 2018: Molecular and Biochemical Parasitology
Luciana Baroni, Luiz M Pereira, Sutherland K Maciver, Ana P Yatsuda
Neospora caninum is an apicomplexan parasite that causes infectious abortion in cows. As an obligate intracellular parasite, N. caninum requires a host cell environment to survive and replicate. The locomotion and invasion mechanisms of apicomplexan parasites are centred on the actin-myosin system to propel the parasite forwards and into the host cell. The functions of actin, an intrinsically dynamic protein, are modulated by actin-binding proteins (ABPs). Actin-depolymerising factor (ADF) is a ubiquitous ABP responsible for accelerating actin turnover in eukaryotic cells and is one of the few known conserved ABPs from apicomplexan parasites...
September 2018: Molecular and Biochemical Parasitology
Cong Liu, Zhenkui Li, Yuanyuan Jiang, Huiting Cui, Jing Yuan
Male and female gametocytes are the infectious forms critical for malaria transmission and targets of intervention. Gametocytes are generally produced in relatively small numbers, and it has been difficult to obtain pure male and female gametocytes for various studies. Male and female gametocytes expressing unique fluorescence reporters have been generated for both Plasmodium falciparum and Plasmodium berghei parasites, which allows isolation of large numbers of pure male and female gametocytes and has greatly contributed to our understanding of gametocyte biology...
September 2018: Molecular and Biochemical Parasitology
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