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New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (31 December 2023) | Viewed by 13007

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Dr. Thiago Almeida Pereira

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Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
Interests: liver fibrosis; host-pathogen interactions; metabolic dysfunction associated steatotic liver disease (MASLD); schistosomiasis; viral hepatitis; biomarkers; drug targets; hedgehog signaling
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Dear Colleagues,

Despite advances in treatment and prevention, diseases caused by parasites are still a significant public health problem worldwide, especially in the tropics. The mechanisms driving the pathogenesis of these life-threatening diseases that disproportionately affect people living in poverty are still not fully elucidated. This gap in knowledge has consequently limited the development of effective treatments (for the infection and the sequela of the disease, such as organ damage), non-invasive biomarkers of infection and morbidity, and improved diagnostic methods. Understanding the host-parasite interactions could help us to dissect the mechanisms underlying disease pathogenesis and develop appropriate interventions to prevent infection and ameliorate or even revert the lesions induced by parasites in chronic infections.

This special issue aims to gather a collection of state-of-the-art and up-to-date articles covering recent advances in host-parasite interactions, the pathogenesis of parasitic diseases, therapeutical interventions, and point-of-care diagnostic tools for active infection and morbidity biomarkers. Submissions of basic and translational original research articles and reviews are welcome. Studies on neglected tropical diseases and host-parasite interactions in the intermediate host are encouraged to be submitted to this special edition.

Dr. Thiago Almeida Pereira
Guest Editor

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Keywords

parasites
host-parasite interactions
immune response
drug targets
diagnosis
biomarkers
vaccines
point-of-care
translational research
neglected tropical diseases

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Published Papers (6 papers)

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Research

16 pages, 1884 KiB

Open AccessArticle

The Development of a One-Step RT-qPCR for the Detection and Quantification of Viable Forms of Trypanosoma cruzi in Açai Samples from Areas at Risk of Chagas Disease through Oral Transmission

by Amanda Faier-Pereira, Paula Finamore-Araujo, Carlos Ramon do Nascimento Brito, Eldrinei Gomes Peres, Klenicy Kazumy de Lima Yamaguchi, Daniele Pereira de Castro and Otacilio C. Moreira

Int. J. Mol. Sci. 2024, 25(10), 5531; https://doi.org/10.3390/ijms25105531 - 18 May 2024

Viewed by 1128

Abstract

Currently, approximately 70% of new cases of Chagas disease (CD) in Brazil are attributed to oral transmission, particularly through foods such as açaí, bacaba, and sugarcane juice, primarily in the northern and northeastern regions of the country. This underscores the imperative need to control the spread of the disease. The methods utilized to conduct quality control for food associated with outbreaks and to assess the potential for the oral transmission of CD through consuming açaí primarily rely on isolating the parasite or inoculating food into experimental animals, restricting the analyses to major research centers. While there are existing studies in the literature on the detection and quantification of T. cruzi DNA in açaí, the evaluation of parasites’ viability using molecular methods in this type of sample and differentiating between live and dead parasites in açaí pulp remain challenging. Consequently, we developed a molecular methodology based on RT-qPCR for detecting and quantifying viable T. cruzi in açaí pulp samples. This protocol enables the stabilization and preservation of nucleic acids in açaí, along with incorporating an exogenous internal amplification control. The standardization of the RNA extraction method involved a simple and reproducible approach, coupled with a one-step RT-qPCR assay. The assay underwent validation with various T. cruzi DTUs and demonstrated sensitivity in detecting up to 0.1 viable parasite equivalents/mL in açaí samples. Furthermore, we investigated the effectiveness of a bleaching method in eliminating viable parasites in açaí samples contaminated with T. cruzi by comparing the detection of DNA versus RNA. Finally, we validated this methodology using açaí pulp samples positive for T. cruzi DNA, which were collected in a municipality with a history of oral CD outbreaks (Coari-AM). This validation involved comparing the detection and quantification of total versus viable T. cruzi. Collectively, our findings demonstrate the feasibility of this methodology in detecting viable forms of T. cruzi in açaí pulp samples, emerging as a crucial tool for monitoring oral outbreaks of Chagas disease resulting from açaí consumption. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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18 pages, 6905 KiB

Open AccessArticle

Characterization of Novel Trypanosoma cruzi-Specific Antigen with Potential Use in the Diagnosis of Chagas Disease

by Micaela S. Ossowski, Juan Pablo Gallardo, Leticia L. Niborski, Jessica Rodríguez-Durán, Walter J. Lapadula, Maximiliano Juri Ayub, Raúl Chadi, Yolanda Hernandez, Marisa L. Fernandez, Mariana Potenza and Karina A. Gómez

Int. J. Mol. Sci. 2024, 25(2), 1202; https://doi.org/10.3390/ijms25021202 - 18 Jan 2024

Viewed by 1755

Abstract

Chagas disease is caused by the parasite Trypanosoma cruzi. In humans, it evolves into a chronic disease, eventually resulting in cardiac, digestive, and/or neurological disorders. In the present study, we characterized a novel T. cruzi antigen named Tc323 (TcCLB.504087.20), recognized by a single-chain monoclonal antibody (scFv 6B6) isolated from the B cells of patients with cardiomyopathy related to chronic Chagas disease. Tc323, a ~323 kDa protein, is an uncharacterized protein showing putative quinoprotein alcohol dehydrogenase-like domains. A computational molecular docking study revealed that the scFv 6B6 binds to an internal domain of Tc323. Immunofluorescence microscopy and Western Blot showed that Tc323 is expressed in the main developmental forms of T. cruzi, localized intracellularly and exhibiting a membrane-associated pattern. According to phylogenetic analysis, Tc323 is highly conserved throughout evolution in all the lineages of T. cruzi so far identified, but it is absent in Leishmania spp. and Trypanosoma brucei. Most interestingly, only plasma samples from patients infected with T. cruzi and those with mixed infection with Leishmania spp. reacted against Tc323. Collectively, our findings demonstrate that Tc323 is a promising candidate for the differential serodiagnosis of chronic Chagas disease in areas where T. cruzi and Leishmania spp. infections coexist. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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14 pages, 3937 KiB

Open AccessArticle

Characterization of the Effect of N-(2-Methoxyphenyl)-1-methyl-1H-benzimidazol-2-amine, Compound 8, against Leishmania mexicana and Its In Vivo Leishmanicidal Activity

by Rocío Nieto-Meneses, Rafael Castillo, Alicia Hernández-Campos, Benjamín Nogueda-Torres, Edgar Oliver López-Villegas, Adriana Moreno-Rodríguez, Félix Matadamas-Martínez and Lilián Yépez-Mulia

Int. J. Mol. Sci. 2024, 25(1), 659; https://doi.org/10.3390/ijms25010659 - 4 Jan 2024

Viewed by 1368

Abstract

Chemotherapy currently available for leishmaniasis treatment has many adverse side effects and drug resistance. Therefore, the identification of new targets and the development of new drugs are urgently needed. Previously, we reported the synthesis of a N-(2-methoxyphenyl)-1-methyl-1H-benzimidazol-2-amine, named compound 8, with an IC₅₀ value in the micromolar range against L. mexicana, it also inhibited 68.27% the activity of recombinant L. mexicana arginase. Herein, we report studies carried out to characterize the mechanism of action of compound 8, as well as its in vivo leishmanicidal activity. It was shown in our ultrastructural studies that compound 8 induces several changes, such as membrane blebbing, the presence of autophagosomes, membrane detachment and mitochondrial and kinetoplast disorganization, among others. Compound 8 triggers the production of ROS and parasite apoptosis. It reduced 71% of the parasite load of L. mexicana in an experimental model of cutaneous leishmaniasis in comparison with a control. Altogether, the data obtained suggest the potential use of compound 8 in the treatment of cutaneous leishmaniasis. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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16 pages, 3080 KiB

Open AccessArticle

Antileishmanial Activity, Toxicity and Mechanism of Action of Complexes of Sodium Usnate with Lanthanide Ions: Eu(III), Sm(III), Gd(III), Nd(III), La(III) and Tb(III)

by Fernanda da Silva, Yasmin Silva Rizk, Amarith Rodrigues das Neves, Estela Mariana Guimarães Lourenço, Alda Maria Teixeira Ferreira, Melquisedeque Mateus Monteiro, Dênis Pires de Lima, Renata Trentin Perdomo, Iluska Senna Bonfá, Mônica Cristina Toffoli-Kadri, Adriana Pereira Duarte, Daniel Mendes Nunes, Marco Antonio Utrera Martines, Eliane Mattos Piranda and Carla Cardozo Pinto de Arruda

Int. J. Mol. Sci. 2024, 25(1), 413; https://doi.org/10.3390/ijms25010413 - 28 Dec 2023

Viewed by 1279

Abstract

Leishmaniases are neglected diseases with limited therapeutic options. Diffuse cutaneous leishmaniasis can occur in Brazil due to Leishmania amazonensis. This study details the antileishmanial activity and cytotoxicity of complexes of sodium usnate (SAU) with lanthanide ions ([LnL₃ (H₂O)_x] (Ln = La(III), Nd(III), Gd(III), Tb(III), Eu(III) and Sm(III); L = SAU). All lanthanide complexes were highly active and more potent than SAU against L. amazonensis promastigotes and intracellular amastigotes (Pro: IC₅₀ < 1.50 μM; Ama: IC₅₀ < 7.52 μM). EuL₃·3H₂O and NdL₃·3H₂O were the most selective and effective on intracellular amastigotes, with a selectivity index of approximately 7.0. In silico predictions showed no evidence of mutagenicity, tumorigenicity or irritation for all complexes. Treatment with EuL₃·3H₂O triggered NO release even at the lowest concentration, indicating NO production as a mechanism of action against the parasite. Incubating promastigotes with the lanthanide complexes, particularly with SmL₃·4H₂O and GdL₃·3H₂O, led to a change in the mitochondrial membrane potential, indicating the ability of these complexes to target this essential organelle. The same complexes caused cell death through cell membrane disruption, but their relationship with early or late apoptotic processes remains unclear. Thus, the inclusion of lanthanide ions in SAU improves selectivity with a promising mechanism of action targeting the mitochondria. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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20 pages, 15794 KiB

Open AccessArticle

Role of the IL-33/ST2 Activation Pathway in the Development of the Hepatic Fibrosis Induced by Schistosoma mansoni Granulomas in Mice

by Laura Maggi, Genil Mororó Araújo Camelo, Izabella Chrystina Rocha, William Pereira Alves, João Marcelo Peixoto Moreira, Thiago Almeida Pereira, Wagner Luiz Tafuri, Élida Mara Leite Rabelo, Ary Correa, Jr., Roselene Ecco and Deborah Aparecida Negrão-Corrêa

Int. J. Mol. Sci. 2023, 24(12), 10237; https://doi.org/10.3390/ijms241210237 - 16 Jun 2023

Cited by 2 | Viewed by 2061

Abstract

Schistosoma mansoni eggs retained in host tissues induce innate cytokine release, contributing to the induction of Type-2 immune responses and granuloma formation, important to restrain cytotoxic antigens, but leading to fibrosis. Interleukin(IL)-33 participates in experimental models of inflammation and chemically induced fibrosis, but its role in S. mansoni-induced fibrosis is still unknown. To explore the role of the IL-33/suppressor of the tumorigenicity 2 (ST2) pathway, serum and liver cytokine levels, liver histopathology, and collagen deposition were comparatively evaluated in S. mansoni-infected wild-type (WT) and IL-33-receptor knockout (ST2^−/−) BALB/c mice. Our data show similar egg counts and hydroxyproline in the livers of infected WT and ST2^−/− mice; however, the extracellular matrix in ST2^−/− granulomas was loose and disorganised. Pro-fibrotic cytokines, such as IL-13 and IL-17, and the tissue-repairing IL-22 were significantly lower in ST2^−/− mice, especially in chronic schistosomiasis. ST2^−/− mice also showed decreased α-smooth muscle actin (α-SMA) expression in granuloma cells, in addition to reduced Col III and Col VI mRNA levels and reticular fibres. Therefore, IL-33/ST2 signalling is essential for tissue repairing and myofibroblast activation during S. mansoni infection. Its disruption results in inappropriate granuloma organisation, partly due to the reduced type III and VI collagen and reticular fibre formation. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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15 pages, 3447 KiB

Open AccessArticle

Development of a Duplex LAMP Assay with Probe-Based Readout for Simultaneous Real-Time Detection of Schistosoma mansoni and Strongyloides spp. -A Laboratory Approach to Point-Of-Care

by Beatriz Crego-Vicente, Pedro Fernández-Soto, Juan García-Bernalt Diego, Begoña Febrer-Sendra and Antonio Muro

Int. J. Mol. Sci. 2023, 24(1), 893; https://doi.org/10.3390/ijms24010893 - 3 Jan 2023

Cited by 6 | Viewed by 4396

Abstract

Loop-mediated isothermal amplification (LAMP) is the most popular technology for point-of-care testing applications due its rapid, sensitive and specific detection with simple instrumentation compared to PCR-based methods. Many systems for reading the results of LAMP amplifications exist, including real-time fluorescence detection using fluorophore-labelled probes attached to oligonucleotide sequences complementary to the target nucleic acid. This methodology allows the simultaneous detection of multiple targets (multiplexing) in one LAMP assay. A method for multiplexing LAMP is the amplification by release of quenching (DARQ) technique by using a 5′-quencher modified LAMP primer annealed to 3′-fluorophore-labelled acting as detection oligonucleotide. The main application of multiplex LAMP is the rapid and accurate diagnosis of infectious diseases, allowing differentiation of co-infecting pathogens in a single reaction. Schistosomiasis, caused among other species by Schistosoma mansoni and strongyloidiasis, caused by Strongyloides stercoralis, are the most common helminth-parasite infections worldwide with overlapping distribution areas and high possibility of coinfections in the human population. It would be of great interest to develop a duplex LAMP to detect both pathogens in the same reaction. In this study, we investigate the use of our two previously developed and well-stablished LAMP assays for S. mansoni and Strongyloides spp. DNA detection in a new duplex real-time eight-primer system based on a modified DARQ probe method that can be performed in a portable isothermal fluorimeter with minimal laboratory resources. We also applied a strategy to stabilize the duplexed DARQ-LAMP mixtures at room temperature for use as ready-to-use formats facilitating analysis in field settings as point-of-care diagnostics for schistosomiasis and strongyloidiasis. Full article

(This article belongs to the Special Issue New Strategies for the Diagnosis and Treatment of Diseases Caused by Parasites)

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