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Molecular and Biochemical Parasitology

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
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
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...
January 22, 2018: Molecular and Biochemical Parasitology
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...
January 18, 2018: Molecular and Biochemical Parasitology
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...
January 10, 2018: Molecular and Biochemical Parasitology
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.falciparum...
January 6, 2018: Molecular and Biochemical Parasitology
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...
January 4, 2018: Molecular and Biochemical Parasitology
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...
January 3, 2018: Molecular and Biochemical Parasitology
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...
December 29, 2017: Molecular and Biochemical Parasitology
Haitham Elaadli, Inyoung Kim, Zachary B Mackey
No abstract text is available yet for this article.
December 26, 2017: Molecular and Biochemical Parasitology
Ewa Harasimczuk, Aina-Cathrine Øvergård, Sindre Grotmol, Frank Nilsen, Sussie Dalvin
The salmon louse, Lepeophtheirus salmonis (Copepoda: Caligidae), is currently the most significant pathogen affecting the salmon farming industry in the Northern Hemisphere. Exocrine glands of blood-feeding parasites are believed to be important for the host-parasite interaction, but also in the production of substances for integument lubrication and antifouling. In L. salmonis; however, we have limited knowledge about the exocrine glands. The aim of this study was therefore to examine three genes containing fibronectin type II (FNII) domains expressed in L...
December 12, 2017: Molecular and Biochemical Parasitology
Alejandra P Frasch, León A Bouvier, Florencia M Oppenheimer, Maria Aparecida Juliano, Luiz Juliano, Adriana K Carmona, Juan José Cazzulo, Gabriela T Niemirowicz
Metallocarboxypeptidases (MCPs) of the M32 family, while broadly distributed among prokaryotic organisms, have so far been only found in a few eukaryotes including trypanosomatids. Among these organisms are human and animal pathogens of medical relevance such as Trypanosoma brucei and Trypanosoma cruzi, the respective causative agents of sleeping sickness and Chagas disease. The M32 MCP orthologues found in these parasites share 72% protein sequence identity. They also present a cytosolic localization, a similar pattern of expression and a marked preference for Arg/Lys residues at P1'...
December 12, 2017: Molecular and Biochemical Parasitology
Ashwini Kumar Ray, Sarah Naiyer, Shashi Shekhar Singh, Alok Bhattacharya, Sudha Bhattacharya
Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) is a versatile sequence independent method to probe RNA structure in vivo and in vitro. It has so far been tried mainly with model organisms. We show that cells of Entamoeba histolytica, a protozoan parasite of humans are hyper-sensitive to the in vivo SHAPE reagent, NAI, and show rapid loss of viability and RNA integrity. We optimized treatment conditions with 5.8S rRNA and Eh_U3 snoRNA to obtain NAI-modification while retaining RNA integrity...
November 22, 2017: Molecular and Biochemical Parasitology
Hiren Banerjee, Richard A Rachubinski
The kinetoplastid parasites Trypanosoma and Leishmania are etiologic agents of diseases like African sleeping sickness, Chagas and leishmaniasis that inflict many tropical and subtropical parts of the world. These parasites are distinctive in that they compartmentalize most of the usually cytosolic enzymes of the glycolytic pathway within a peroxisome-like organelle called the glycosome. Functional glycosomes are essential in both the procyclic and bloodstream forms of trypanosomatid parasites, and mislocalization of glycosomal enzymes to the cytosol is fatal for the parasite...
December 2017: Molecular and Biochemical Parasitology
Brenda H Welter, Michael G Sehorn, Lesly A Temesvari
Entamoeba histolytica causes dysentery and liver abscess mostly in countries that lack proper sanitation. Infection is acquired by ingestion of the cyst form in contaminated food or water. E. histolytica does not encyst in vitro; thus, E. invadens, a reptilian parasite that encysts in vitro, has been used as a surrogate. Cysts are small and possess chitin-rich walls. These are characteristics that may be exploited by flow cytometry. We stained encysting E. invadens cells with a fluorescent chitin stain, and analyzed fluorescence and forward scatter by flow cytometry...
December 2017: Molecular and Biochemical Parasitology
Roberto R Moraes Barros, Tyler J Gibson, Whitney A Kite, Juliana M Sá, Thomas E Wellems
Human infections from Plasmodium knowlesi present challenges to malaria control in Southeast Asia. P. knowlesi also offers a model for other human malaria species including Plasmodium vivax. P. knowlesi parasites can be cultivated in the laboratory, and their transformation is standardly performed by direct electroporation of schizont-infected red blood cells (RBCs) with plasmid DNA. Here we show that the efficiency of direct electroporation is exquisitely dependent on developmental age of the schizonts. Additionally, we show that transformation of P...
December 2017: Molecular and Biochemical Parasitology
Ebony Weems, Ujjal K Singha, Joseph T Smith, Minu Chaudhuri
Trypanosoma brucei Tim17(TbTim17), the single member of the Tim17/23/22 protein family, is an essential component of the translocase of the mitochondrial inner membrane (TIM). In spite of the conserved secondary structure, the primary sequence of TbTim17, particularly the N-terminal hydrophilic region, is significantly divergent. In order to understand the function of this region we expressed two N-terminal deletion mutants (Δ20 and Δ30) of TbTim17 in T. brucei. Both of these mutants of TbTim17 were targeted to mitochondria, however, they failed to complement the growth defect of TbTim17 RNAi cells...
December 2017: Molecular and Biochemical Parasitology
Alexander K Brazenor, David S Francis, Kate S Hutson, Alexander G Carton
This study on the eggs of the tropical monogenean Neobenedenia girellae presents the first detailed quantitative biochemical information of a marine parasite species' eggs. Moisture and protein composed the majority of the contents of freshly laid eggs (79.12±0.82 and 11.51±0.49% respectively) followed by lipid (2.50±0.15%). Lipids were composed of approximately equal amounts of saturated, monounsaturated, and polyunsaturated fatty acids and the predominant lipid class was triacylglycerol (33.82±1.20%)...
December 2017: Molecular and Biochemical Parasitology
Gabriela Maggioli, Gabriel Rinaldi, Ines Giaudrone, Patricia Berasain, José F Tort, Paul J Brindley, Carlos Carmona
Schistosomiasis is a major neglected tropical disease (NTD) and considered the most important of the human helminthiases in terms of morbidity and mortality. Whereas treatment with praziquantel has been effective since the 1980s, the potential for the emergence of drug resistance has propelled the search for new interventions. Studies have revealed key roles of proteases in parasitic helminths during establishment of infection, tissue invasion, immune evasion, parasite feeding and development throughout the different developmental stages, pinpointing them as possible candidates...
November 20, 2017: Molecular and Biochemical Parasitology
Bornali Deb, Arif Uddin, Gulshana Akthar Mazumder, Supriyo Chakraborty
The phenomenon of unequal usage of synonymous codons encoding an amino acid in which some codons are more preferred to others is the codon usage bias (CUB) and it is species specific. Analysis of CUB helps in understanding evolution at molecular level and acquires significance in mRNA translation, design of transgenes and new gene discovery. In our current study, we analyzed synonymous codon usage pattern and the factors influencing it on mitochondrial protein coding genes of 6 different hookworms i.e. Ancylostoma ceylanicum, Ancylostoma duodenale, Necator americanus, Ancylostoma tubaeforme, Ancylostoma caninum and Uncinaria sanguinis as no work was reported yet...
November 17, 2017: Molecular and Biochemical Parasitology
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