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Molecular and Biochemical Parasitology

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https://www.readbyqxmd.com/read/29684399/a-cas9-transgenic-plasmodium-yoelii-parasite-for-efficient-gene-editing
#1
Pengge Qian, Xu Wang, Zhenke Yang, Zhenkui Li, Han Gao, Xin-Zhuan Su, Huiting Cui, Jing Yuan
The RNA-guided endonuclease Cas9 has applied as an efficient gene-editing method in malaria parasite Plasmodium. However, the size (4.2 kb) of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for genome editing in the parasites only introduced with cas9 plasmid. To establish the endogenous and constitutive expression of Cas9 protein in the rodent malaria parasite P. yoelii, we replaced the coding region of an endogenous gene sera1 with the intact SpCas9 coding sequence using the CRISPR/Cas9-mediated genome editing method, generating the cas9-knockin parasite (PyCas9ki) of the rodent malaria parasite P...
April 20, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29679605/circulating-cell-free-dna-in-patients-with-alveolar-echinococcosis
#2
Alice Baraquin, Eric Hervouet, Carine Richou, Pierre Flori, Paul Peixoto, Amel Azizi, Eric Delabrousse, Oleg Blagosklonov, Gerald Umhang, Solange Bresson-Hadni, Benoit Valot, Frédéric Grenouillet
Alveolar echinococcosis (AE) is a parasitic disease, due to Echinococcus multilocularis. Often compared to liver cancer, it develops by infiltration from its primary site to the surrounding tissue, and can then metastasize to other organs. Detection of circulating cell-free DNA (ccfDNA) is a useful analytical tool in oncology, for diagnosis, prognosis, and therapy monitoring. This study sought to investigate the presence of ccfDNA in patients with AE, and its potential usefulness for the evaluation of treatment efficiency...
April 18, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29655799/stage-specific-expression-of-the-proline-alanine-transporter-in-the-human-pathogen-leishmania
#3
T Liburkin-Dan, D Schlisselberg, R Fischer-Weinberger, P Pescher, E Inbar, M Ephros, D Rentsch, G F Späth, D Zilberstein
Leishmania are obligatory intracellular parasites that cycle between the sand fly midgut (extracellular promastigotes) and mammalian macrophage phagolysosomes (intracellular amastigotes). They have developed mechanisms of adaptation to the distinct environments of host and vector that favor utilization of both proline and alanine. LdAAP24 is the L. donovani proline-alanine transporter. It is a member of Leishmania system A that translocates neutral amino acids. Since system A is promastigote-specific, we aimed to assess whether LdAAP24 is also expressed exclusively in promastigotes...
April 12, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29625152/g-protein-coupled-receptor-genes-of-dirofilaria-immitis
#4
Thangadurai Mani, Catherine Bourguinat, Roger K Prichard
The diversity and uniqueness of nematode heterotrimeric G-protein-coupled receptors (GPCRs) provides impetus for identifying ligands that can be used as therapeutics for treating diseases caused by parasitic nematode infections. In human medicine, GPCRs have represented the largest group of 'drugable' targets exploited in the market today. In the filarial nematode Dirofilaria immitis, which causes heartworm disease, the macrocyclic lactones (ML) have been used as the sole preventatives for more than 25 years and now there is confirmed ML resistance in this parasite...
April 3, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29581012/the-pseudo-dimeric-tyrosyl-trna-synthetase-of-t-brucei-aminoacylates-cytosolic-and-mitochondrial-trna-tyr-and-requires-both-monomeric-units-for-activity
#5
Sandro Käser, Isabelle Glauser, Jochen Rettig, André Schneider
Aminoacyl-tRNA synthetases are essential for protein synthesis. The single-copy tyrosyl-tRNA synthetase (Tb-TyrRS) of T. brucei has an unusual structure and forms a pseudo-dimer. It is therefore twice the size than tyrosyl-tRNA synthetases of most other organisms. Here we show by inducible RNAi that Tb-TyrRS is essential for normal growth of procyclic T. brucei. Furthermore we demonstrate that Tb-TyrRS aminoacylates cytosolic as well as mitochondrial tRNATyr indicating that it is dually localized. Finally we show that individual deletion of the 36 N- or C-terminal amino acids abolishes the function of Tb-TyrRS...
March 23, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29581011/a-plant-like-mitochondrial-carrier-family-protein-facilitates-mitochondrial-transport-of-di-and-tricarboxylates-in-trypanosoma-brucei
#6
Claudia Colasante, Fuli Zheng, Cordula Kemp, Frank Voncken
The procyclic form of the human parasite Trypanosoma brucei harbors one single, large mitochondrion containing all tricarboxylic acid (TCA) cycle enzymes and respiratory chain complexes present also in higher eukaryotes. Metabolite exchange among subcellular compartments such as the cytoplasm, the mitochondrion, and the peroxisomes is crucial for redox homeostasis and for metabolic pathways whose enzymes are dispersed among different organelles. In higher eukaryotes, mitochondrial carrier family (MCF) proteins transport TCA-cycle intermediates across the inner mitochondrial membrane...
March 23, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29581010/molecular-cloning-expression-and-characterization-of-udp-n-acetyl-%C3%AE-d-galactosamine-polypeptide-n-acetylgalactosaminyltransferase-4-from-cryptosporidium-parvum
#7
Maria A DeCicco RePass, Najma Bhat, Jamie Heimburg Molinaro, Stephen Bunnell, Richard D Cummings, Honorine D Ward
Cryptosporidium spp. are the causative agents of diarrheal disease worldwide, but effective treatments are lacking. Cryptosporidium employs mucin-like glycoproteins with O-glycans to attach to and infect host intestinal epithelial cells. The Tn antigen (GalNAcα1-Ser/Thr) is an O-glycan essential for these processes, as Tn-specific lectins and a Tn-specific monoclonal antibody block attachment to and infection of host cells in vitro. The enzymes in Cryptosporidium catalyzing their synthesis, however, have not been studied...
March 23, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29555232/heterologous-expression-of-three-antigenic-proteins-from-angiostrongylus-cantonensis-es-7-lec-5-and-14-3-3-in-mammalian-cells
#8
Bianca B Cognato, Sukwan Handali, Alessandra L Morassutti, Alexandre Januário da Silva, Carlos Graeff-Teixeira
Angiostrongylus cantonensis is a parasitic nematode and the main causative agent of human cerebral eosinophilic meningoencephalitis (EoM). A definitive diagnosis of EoM usually requires serologic or molecular analysis of the patient's clinical sample. Currently, a 31 kDa antigen is used in immunological tests for this purpose, however as a crude antigen preparation it may present cross-reactivity with other helminthic infections, especially echinococcosis. Heterologous expression studies using prokaryotic systems failed on producing antigenic proteins...
March 16, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29477861/schistosoma-mansoni-venom-allergen-like-protein-18-smval18-is-a-plasminogen-binding-protein-secreted-during-the-early-stages-of-mammalian-host-infection
#9
Rafaela S Fernandes, Luis G V Fernandes, Andre S de Godoy, Patrícia A Miyasato, Eliana Nakano, Leonardo P Farias, Ana L T O Nascimento, Luciana C C Leite
Schistosomiasis is a neglected tropical disease caused by trematodes of the genus Schistosoma which have a complex life cycle characterized by an asexual multiplication in the snail intermediate host and a sexual reproduction phase in the mammalian definitive host. The initial steps of the human host infection involve the secretion of proteins contained in the acetabular glands of cercariae that promote parasite adhesion and proteolysis of the skin layers. Herein, we performed a functional analysis of SmVAL18, identified as one of the three SCP/TAPS proteins constituent of cercarial secretions...
February 22, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29453993/the-tegumental-allergen-like-proteins-of-schistosoma-mansoni-a-biochemical-study-of-smtal4-tal13
#10
Jack Carson, Charlotte M Thomas, Aaron McGinty, Gustavo Takata, David J Timson
Schistosoma mansoni, like other trematodes, expresses a number of unusual calcium binding proteins which consist of an EF-hand domain joined to a dynein light chain-like (DLC-like) domain by a flexible linker. These proteins have been implicated in host immune responses and drug binding. Three members of this protein family from S. mansoni (SmTAL1, SmTAL2 and SmTAL3) have been well characterised biochemically. Here we characterise the remaining family members from this species (SmTAL4-13). All of these proteins form homodimers and all except SmTAL5 bind to calcium and manganese ions...
February 14, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29414671/fibrinogen-and-fibrin-are-novel-substrates-for-fasciola-hepatica-cathepsin-l-peptidases
#11
Mirjam M Mebius, Jody M J Op Heij, Aloysius G M Tielens, Philip G de Groot, Rolf T Urbanus, Jaap J van Hellemond
Cathepsin peptidases form a major component of the secreted proteins of the blood-feeding trematodes Fasciola hepatica and Schistosoma mansoni. These peptidases fulfill many functions, from facilitating infection to feeding and immune evasion. In this study, we examined the Fasciola cathepsin L peptidases FhCL1, FhCL2, and FhCL3 and the schistosomal cathepsin peptidases SmCB1 and SmCL3 for their anticoagulant properties. Although no direct anticoagulant effect of these peptidases was observed, we discovered that cathepsin peptidases from Fasciola, but not from Schistosoma, were able to degrade purified fibrinogen, with FhCL1 having the highest fibrinogenolytic activity...
February 4, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29409763/knockout-of-the-ccch-zinc-finger-protein-tczc3h31-blocks-trypanosoma-cruzi-differentiation-into-the-infective-metacyclic-form
#12
Monica Visnieski Alcantara, Rafael Luis Kessler, Rosana Elisa Gonçalves Gonçalves, Newmar Pinto Marliére, Alessandra Aparecida Guarneri, Gisele Fernanda Assine Picchi, Stenio Perdigão Fragoso
In the protozoan parasite Trypanosoma cruzi - the causative agent of Chagas disease - gene expression control is mainly post-transcriptional, where RNA-binding proteins (RBPs) play a central role, by controlling mRNA stability, distribution and translation. A large variety of RBPs are encoded in the T. cruzi genome, including the CCCH-type zinc finger (CCCH ZnF) protein family, which is characterized by the presence of the C-X7/8 -C-X5 -C-X3 -H (CCCH) motif. In the related parasite T. brucei, CCCH ZnF proteins have been shown to control key differentiation steps in the parasite's life cycle...
February 2, 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29371154/trypanosoma-brucei-bloodstream-forms-express-highly-specific-and-separate-transporters-for-adenine-and-hypoxanthine-evidence-for-a-new-protozoan-purine-transporter-family
#13
Gustavo D Campagnaro, Khalid J Alzahrani, Jane C Munday, Harry P De Koning
The transport of nucleobases and nucleosides in protozoan parasites is known to be performed by Equilibrative Nucleoside Transporter (ENT) family members, including the extensively studied P1 and P2 nucleoside transporters of T. brucei bloodstream forms. Studies with P2 knockout parasites suggested the existence of as yet uncharacterised purine transport mechanisms in these cells. Here, we deleted several ENT genes, in addition to P2, including an array comprising three genes encoding for high-affinity broad-selectivity nucleobase transporters - the longest multi-gene locus deletion in T...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29357296/spontaneous-excision-and-facilitated-recovery-as-a-control-for-phenotypes-arising-from-rna-interference-and-other-dominant-transgenes
#14
Erin A Brettmann, Lon-Fye Lye, Stephen M Beverley
An essential control for genetic manipulation of microbes is the regeneration of the wild-type state and phenotype to validate that any mutant phenotypes are 'on target'. For Leishmania gene knockouts, this is often done by re-expression of the target gene from episomal vectors, often bearing counter-selectable markers. Methods for similarly validating the outcomes from dominant mutations such as those arising from RNA interference (RNAi) are needed. We present here such an approach, relying on facilitated recovery after spontaneous excision - or 'popouts' - of dominant transgenes stably inserted into the ribosomal RNA array, utilizing GFP as a marker and single cell sorting to recover regenerated WT controls...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29331577/a-novel-encystation-specific-protein-kinase-regulates-chitin-synthesis-in-entamoeba-invadens
#15
Sintu Kumar Samanta, Sneha Susan Varghese, Deepak Krishnan, Mithu Baidya, Digant Nayak, Sumanta Mukherjee, Sudip K Ghosh
Phosphorylation is an important post-translational modification of proteins and is involved in the regulation of a variety of cellular events. The proteome of Entamoeba invadens, the reptilian counterpart of Entamoeba histolytica consists of an overwhelming number of putative protein kinases, and some may have a role to play in Entamoeba encystation. In this study, we have identified a novel protein kinase named as EiCSpk (Entamoeba invadenscyst specific protein kinase) which expressed almost exclusively during encystation...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29317266/biochemical-characterization-of-unusual-cysteine-protease-of-p-falciparum-metacaspase-2-mca-2
#16
Vandana, Agam P Singh, Jitendra Singh, Ruby Sharma, Mymoona Akhter, Pradyumna K Mishra, Ajay K Saxena, Rajnikant Dixit, Brijesh Rathi, Anju Katyal, Kailash C Pandey
Earlier studies on Plasmodium apoptosis revealed the presence of proteases with caspases like- activity, which are known as "metacaspases". Although this family of cysteine proteases is structurally similar to caspases with Cys-His dyad but their evolutionary significance and functional relevance remains largely unknown. These proteases are considered to be an important target against malaria due to their absence in humans. In this report, we have biochemically characterized metacaspase-2 (PfMCA-2) of P...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29307698/sequence-and-functional-divergence-of-gametocyte-specific-parasitophorous-vacuole-membrane-proteins-in-plasmodium-parasites
#17
Elena Deligianni, Maria Andreadaki, Konstantinos Koutsouris, Inga Siden-Kiamos
Plasmodium parasites develop within red blood cells in a Parasitophorous Vacuole enclosed by a Membrane, the PVM. The protein family ETRAMP (Early Transcribed Membrane Protein) comprises small proteins inserted in the PVM via a single transmembrane domain. Among those, Pfs16 is specifically found in P. falciparum gametocyte PVM. The P. berghei gene PBANKA_1003900 is syntenic with pfs16. The encoded proteins have a similar domain structure but the overall protein similarity is low. A transcript of the P. berghei gene is only found in gametocytes and ookinetes and a C-terminal mCherry fusion of the protein revealed its presence only in gametocytes...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29305880/distinct-effects-of-hiv-protease-inhibitors-and-erad-inhibitors-on-zygote-to-ookinete-transition-of-the-malaria-parasite
#18
Evi Goulielmaki, Sofia Kaforou, Kannan Venugopal, Thanasis G Loukeris, Inga Siden-Kiamos, Konstantinos Koussis
In an effort to eradicate malaria, new interventions are proposed to include compound/vaccine development against pre-erythrocytic, erythrocytic and mosquito stages of Plasmodium. Drug repurposing might be an alternative approach to new antimalarials reducing the cost and the time required for drug development. Previous in vitro studies have examined the effects of protease inhibitors on different stages of the Plasmodium parasite, although the clinical relevance of this remains unclear. In this study we tested the putative effect of three HIV protease inhibitors, two general aspartyl protease inhibitors and three AAA-p97 ATPase inhibitors on the zygote to ookinete transition of the Plasmodium parasite...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29292212/enrichment-of-cryptosporidium-parvum-from-in-vitro-culture-as-measured-by-total-rna-and-subsequent-sequence-analysis
#19
Jessica C Kissinger, Karen E Hermetz, Keith M Woods, Steve J Upton
Cryptosporidium parvum is an apicomplexan parasite that infects a wide range of hosts including humans. Due to the parasite's quasi-intracellular, intermembrane location on the host cell, it is difficult to purify parasites from in vitro and in vivo infections for molecular studies. We have developed a method to greatly enrich in vitro C. parvum merozoites from host cells. The efficiency of the protocol was assessed with C. parvum (KSU-1 isolate) parasites of different developmental stages isolated following a synchronized infection of HCT-8 host cells...
March 2018: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29287675/depletion-of-the-extracellular-signal-regulated-kinase-8-homolog-in-trypanosoma-brucei-in-vivo-reduces-its-virulence-in-a-mouse-target-validation-study
#20
Haitham Elaadli, Inyoung Kim, Zachary B Mackey
Trypanosoma brucei sub-species are vector borne kinetoplastid parasites that cause the potentially lethal disease Human African trypanosomiasis. The target-based therapy for curing this parasitic disease relies on one drug, Eflornithine. The roles of mitogen-activated protein kinases in regulating key cellular processes in eukaryotic cells such as proliferation, stress response and differentiation plus their druggability make them attractive targets for therapeutic exploitation. The extracellular-regulated kinase 8 homolog in T...
March 2018: Molecular and Biochemical Parasitology
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