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Molecular and Biochemical Parasitology

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https://www.readbyqxmd.com/read/28645481/subcellular-localization-of-glycolytic-enzymes-and-characterization-of-intermediary-metabolism-of-trypanosoma-rangeli
#1
Rocío Rondón-Mercado, Héctor Acosta, Ana J Cáceres, Wilfredo Quiñones, Juan Luis Concepción
Trypanosoma rangeli is a hemoflagellate protist that infects wild and domestic mammals as well as humans in Central and South America. Although this parasite is not pathogenic for human, it is being studied because it shares with Trypanosoma cruzi, the etiological agent of Chagas' disease, biological characteristics, geographic distribution, vectors and vertebrate hosts. Several metabolic studies have been performed with T. cruzi epimastigotes, however little is known about the metabolism of T. rangeli. In this work we present the subcellular distribution of the T...
June 20, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28633997/nicotinamide-inhibits-the-growth-of-p-falciparum-and-enhances-the-antimalarial-effect-of-artemisinin-chloroquine-and-pyrimethamine
#2
Sergey O Tcherniuk, Olga Chesnokova, Irina V Oleinikov, Andrew V Oleinikov
Nicotinamide (vitamin B3) - is a water soluble amide derivative of nicotinic acid, which has been used at high doses for a variety of therapeutic applications. However, its antimalarial effect has not been intensively explored. In this work we analysed the effect of nicotinamide alone and in combination with artemisinin, chloroquine andpyrimethamine on the growth of blood stages of P. falciparum. Our results demonstrate that nicotinamide effectively inhibits the growth of blood stage parasites with IC50 of 6...
June 17, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28629935/a-dicre-recombinase-based-system-for-inducible-expression-in-leishmania-major
#3
Renato E R S Santos, Gabriel L A Silva, Elaine V Santos, Samuel M Duncan, Jeremy C Mottram, Jeziel D Damasceno, Luiz R O Tosi
Here we present the establishment of an inducible system based on the dimerizable Cre recombinase (DiCre) for controlled gene expression in the protozoan parasite Leishmania. Rapamycin-induced DiCre activation promoted efficient flipping and expression of gene products in a time and dose-dependent manner. The DiCre flipping activity induced the expression of target genes from both integrated and episomal contexts broadening the applicability of the system. We validated the system by inducing the expression of both full length and truncated forms of the checkpoint protein Rad9, which revealed that the highly divergent C-terminal domain of Rad9 is necessary for proper subcellular localization...
June 16, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28629934/recent-advances-in-leishmania-reverse-genetics-manipulating-a-manipulative-parasite
#4
REVIEW
Samuel M Duncan, Nathaniel G Jones, Jeremy C Mottram
In this review we describe the expanding repertoire of molecular tools with which to study gene function in Leishmania. Specifically we review the tools available for studying functions of essential genes, such as plasmid shuffle and DiCre, as well as the rapidly expanding portfolio of available CRISPR/Cas9 approaches for large scale gene knockout and endogenous tagging. We include detail on approaches that allow the direct manipulation of RNA using RNAi and protein levels via Tet or DiCre induced overexpression and destabilization domain mediated degradation...
June 16, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28627360/the-plasmodium-falciparum-exported-protein-pf3d7_0402000-binds-to-erythrocyte-ankyrin-and-band-4-1
#5
Bikash Shakya, Wesley D Penn, Ernesto S Nakayasu, Douglas J LaCount
Plasmodium falciparum extensively modifies the infected red blood cell (RBC), resulting in changes in deformability, shape and surface properties. These alterations suggest that the RBC cytoskeleton is a major target for modification during infection. However, the molecular mechanisms leading to these changes are largely unknown. To begin to address this question, we screened for exported P. falciparum proteins that bound to the erythrocyte cytoskeleton proteins ankyrin 1 (ANK1) and band 4.1 (4.1R), which form critical interactions with other cytoskeletal proteins that contribute to the deformability and stability of RBCs...
June 13, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28602728/the-fluorescent-protein-ilov-outperforms-egfp-as-a-reporter-gene-in-the-microaerophilic-protozoan-trichomonas-vaginalis
#6
Shuqi E Wang, Anna E S Brooks, Bronwyn Cann, Augusto Simoes-Barbosa
Trichomonas vaginalis is a flagellated protozoan causing a notorious urogenital infection in humans. Due to its anaerobic metabolism, an alternative fluorescent protein that can be readily expressed in oxygen-deprived conditions is ideal. This study assessed the performance of iLOV, which does not require oxygen to function, as compared to the conventional enhanced green fluorescent protein (eGFP) in T. vaginalis. The results indicated that iLOV outperforms eGFP in both transient and stable expression, being detectable earlier and producing higher fluorescent intensity than eGFP in T...
June 9, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28522152/anti-trypanosomal-activity-of-cationic-n-heterocyclic-carbene-gold-i-complexes
#7
Isabel Winter, Julia Lockhauserbäumer, Gertrud Lallinger-Kube, Rainer Schobert, Klaus Ersfeld, Bernhard Biersack
Two gold(I) N-heterocyclic carbene complexes 1a and 1b were tested for their anti-trypanosomal activity against Trypanosoma brucei parasites. Both gold compounds exhibited excellent anti-trypanosomal activity (IC50=0.9-3.0nM). The effects of the gold complexes 1a and 1b on the T. b. brucei cytoskeleton were evaluated. Rapid detachment of the flagellum from the cell body occurred after treatment with the gold complexes. In addition, a quick and complete degeneration of the parasitic cytoskeleton was induced by the gold complexes, only the microtubules of the detached flagellum remained intact...
May 15, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28438458/an-efficient-cumate-inducible-system-for-procyclic-and-bloodstream-form-trypanosoma-brucei
#8
Feng-Jun Li, Zhi-Shen Xu, Htay Mon Aye, Anaïs Brasseur, Zhao-Rong Lun, Kevin S W Tan, Cynthia Y He
In Trypanosoma brucei, the tetracycline-inducible system enables tightly-regulated, highly-efficient expression of recombinant proteins or double-stranded RNA in both procyclic and bloodstream form cells, providing useful molecular genetic tools to study gene functions. An alternative, vanillic acid-inducible system is recently described for procyclic T. brucei, providing ∼18-fold increase in GFP reporter expression upon induction (Sunter JD. Mol. Biochem. Parasitol. 2016, 207:45-48). Here we describe a cumate-inducible system that allows efficient, tunable gene expression showing >300-fold increase in GFP expression upon induction...
April 21, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28427949/parasitism-finds-many-solutions-to-the-same-problems-in-red-algae-florideophyceae-rhodophyta
#9
Jillian M Freese, Christopher E Lane
Parasitic red algae evolve from a common ancestor with their hosts, parasitizing cousins using familiar cellular mechanisms. They have independently evolved over one hundred times within the exclusively multicellular red algal class Florideophyceae. Reduced morphology, a lack of pigmentation, and direct cell-cell connections with their hosts are markers of red algal parasitism. With so many potential evolutionary pathways, red algal parasite diversity offers a unique test case to understand the earliest stages of this lifestyle transition...
April 17, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28414172/lccl-protein-complex-formation-in-plasmodium-is-critically-dependent-on-lap1
#10
Annie Z Tremp, Vikram Sharma, Victoria Carter, Edwin Lasonder, Johannes T Dessens
Successful sporogony of Plasmodium berghei in vector mosquitoes requires expression of a family of six modular proteins named LCCL lectin domain adhesive-like proteins (LAPs). The LAPs share a subcellular localization in the crystalloid, a unique parasite organelle that forms during ookinete development. Here, LAP interactions in P. berghei were studied using a series of parasite lines stably expressing reporter-tagged LAPs combined with affinity purification and high accuracy label free quantitative mass spectrometry...
April 13, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28392476/schistosoma-mansoni-displays-an-adenine-phosphoribosyltransferase-preferentially-expressed-in-mature-female-gonads-and-vitelaria
#11
Ana Eliza Zeraik, Vitor Hugo Balasco Serrão, Larissa Romanello, Juliana Roberta Torini, Alexandre Cassago, Ricardo DeMarco, Humberto D'Muniz Pereira
Schistosoma mansoni depends upon the purine salvage pathway to obtain purine nucleotides; therefore, enzymes from this pathway are essential for parasite survival. Here, we focused on the adenine phosphoribosyltransferase (APRT) enzyme, which catalyzes the condensation reaction between adenine and PRPP (5-phosphoribosylpyrophosphate) to produce AMP and PPi. Kinetic experiments using the heterologously expressed protein of one APRT isoform from S. mansoni indicate that it is catalytically active, and whole-mount in situ hybridization studies indicate that the transcripts of this protein are concentrated in the posterior region of the ovary and vitellaria of female adult worms...
April 6, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28389272/characterization-of-ciliobrevin-a-mediated-dynein-atpase-inhibition-on-flagellar-motility-of-leishmania-donovani
#12
G Srinivas Reddy, Aakash Gautam Mukhopadhyay, Chinmoy Sankar Dey
Axonemal dyneins are members of AAA+ proteins involved in force generation and are responsible for flagellar motility in eukaryotes. In this study, we characterized the effects of ciliobrevin A (CbA), a dynein ATPase inhibitor, on flagella driven motility of the protozoan parasite Leishmania donovani. Using fast-capture video microscopy, we observed that CbA decreased flagellar beat frequency of swimming parasites in a concentration-dependent manner. Beat frequency of live and reactivated L. donovani decreased by approximately 89% and 41% respectively in the presence of 250μM CbA...
April 5, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28385564/genome-wide-identification-of-microrna-targets-in-the-neglected-disease-pathogens-of-the-genus-echinococcus
#13
Natalia Macchiaroli, Lucas L Maldonado, Magdalena Zarowiecki, Marcela Cucher, María Inés Gismondi, Laura Kamenetzky, Mara Cecilia Rosenzvit
MicroRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in biological processes such as development. MiRNAs silence target mRNAs by binding to complementary sequences in the 3'untranslated regions (3'UTRs). The parasitic helminths of the genus Echinococcus are the causative agents of echinococcosis, a zoonotic neglected disease. In previous work, we performed a comprehensive identification and characterization of Echinococcus miRNAs...
April 3, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28385563/evidence-of-putative-non-coding-rnas-from-leishmania-untranslated-regions
#14
Felipe Freitas Castro, Patricia C Ruy, Karina Nogueira Zeviani, Ramon Freitas Santos, Juliano Simões Toledo, Angela Kaysel Cruz
Non-coding RNAs (ncRNAs) are regulatory elements present in a wide range of organisms, including trypanosomatids. ncRNAs transcribed from the untranslated regions (UTRs) of coding genes have been described in the transcriptomes of several eukaryotes, including Trypanosoma brucei. To uncover novel putative ncRNAs in two Leishmania species, we examined a L. major cDNA library and a L. donovani non-polysomal RNA library. Using a combination of computational analysis and experimental approaches, we classified 26 putative ncRNA in L...
April 3, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28373094/inducible-protein-stabilization-system-in-leishmania-mexicana
#15
Lucie Podešvová, Huan Huang, Vyacheslav Yurchenko
Targeted regulation of protein levels is an important tool to investigate the role of proteins essential for cell function and development. In recent years, methods based on the Escherichia coli dihydrofolate reductase destabilization domain (ecDHFR DD) have been established and used in various cell types. ecDHFR DD destabilizes the fused protein of interest and causes its degradation by proteasomes, unless it is stabilized by a specific ligand, trimethoprim. In this work we developed an inducible protein stabilization system in Leishmania mexicana based on ecDHFR DD...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28373093/experimental-and-bioinformatic-characterization-of-cabp2933-an-ef-hand-protein-of-giardia-intestinalis
#16
Magda E Alvarado, Claudia Rubiano, Eliana Calvo, Vanessa Gómez, Moisés Wasserman
Giardia intestinalis is a parasite that inhabits the small intestine of humans. This parasite is a divergent eukaryote with a compact genome. The calcium ion is an essential messenger in cell signaling. Calcium's role as a messenger is mediated through calcium-binding proteins (CaBPs) that decode the message. The most important family of CaBPs is the EF-Hand protein family. In this study we have explored the role of EF-Hand protein CaBP2933. We analyzed its location, confirmed its ability to bind calcium and identified some of its interacting proteins...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28366668/the-intermembrane-space-protein-erv1-of-trypanosoma-brucei-is-essential-for-mitochondrial-fe-s-cluster-assembly-and-operates-alone
#17
Alexander C Haindrich, Michala Boudová, Marie Vancová, Priscila Peña Diaz, Eva Horáková, Julius Lukeš
Sulfhydryl oxidase Erv1 is a ubiquitous and conserved protein of the mitochondrial intermembrane space that plays a role in the transport of small sulfur-containing proteins. In higher eukaryotes, Erv1 interacts with the mitochondrial import protein Mia40. However, Trypanosoma brucei lacks an obvious Mia40 homologue in its genome. Here we show by tandem affinity purification and mass spectrometry that in this excavate protist, Erv1 functions without a Mia40 homologue and most likely any other interaction partner...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28366667/characterization-and-role-of-the-3-methylglutaconyl-coenzyme-a-hidratase-in-trypanosoma-brucei
#18
Mariana Leão de Lima Stein, Marcelo Yudi Icimoto, Erica Valadares de Castro Levatti, Vitor Oliveira, Anita Hilda Straus, Sergio Schenkman
Trypanosoma brucei, the agent of African Trypanosomiasis, is a flagellated protozoan parasite that develops in tsetse flies and in the blood of various mammals. The parasite acquires nutrients such as sugars, lipids and amino acids from their hosts. Amino acids are used to generate energy and for protein and lipid synthesis. However, it is still unknown how T. brucei catabolizes most of the acquired amino acids. Here we explored the role of an enzyme of the leucine catabolism, the 3-methylglutaconyl-Coenzyme A hydratase (3-MGCoA-H)...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28623962/editorial
#19
EDITORIAL
Patrick Skelly, Philip T LoVerde
No abstract text is available yet for this article.
July 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28188804/metabolite-profiling-of-infection-associated-metabolic-markers-of-onchocerciasis
#20
Sasisekhar Bennuru, Sara Lustigman, David Abraham, Thomas B Nutman
The global efforts for onchocerciasis elimination may require additional tools (safe micro and macrofilaricidal drugs, vaccines and biomarkers) as elimination efforts move toward the "end game". Efforts toward the identification of suitable biomarkers have focused on specific protein(s) and/or nucleic acids, but metabolites present an alternative option as they have limited half-lives and are the result of combinatorial effects. In comparison to previously used methodology of LC-MS for metabolomic approaches, we used a non-targeted capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) to analyze the serum metabolic profiles of Ov-infected and -uninfected individuals (n=20)...
July 2017: Molecular and Biochemical Parasitology
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