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1.
ACS Infect Dis ; 2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-39023360

RESUMEN

The lack of effective vaccines and the development of resistance to the current treatments highlight the urgent need for new anti-leishmanials. Sphingolipid metabolism has been proposed as a promising source of Leishmania-specific targets as these lipids are key structural components of the eukaryotic plasma membrane and are involved in distinct cellular events. Inositol phosphorylceramide (IPC) is the primary sphingolipid in the Leishmania species and is the product of a reaction mediated by IPC synthase (IPCS). The antihistamine clemastine fumarate has been identified as an inhibitor of IPCS in L. major and a potent anti-leishmanial in vivo. Here we sought to further examine the target of this compound in the more tractable species L. mexicana, using an approach combining genomic, proteomic, metabolomic and lipidomic technologies, with molecular and biochemical studies. While the data demonstrated that the response to clemastine fumarate was largely conserved, unexpected disturbances beyond sphingolipid metabolism were identified. Furthermore, while deletion of the gene encoding LmxIPCS had little impact in vitro, it did influence clemastine fumarate efficacy and, importantly, in vivo pathogenicity. Together, these data demonstrate that clemastine does inhibit LmxIPCS and cause associated metabolic disturbances, but its primary target may lie elsewhere.

2.
PLoS One ; 19(7): e0306600, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39008475

RESUMEN

Echinococcus spp. is an emerging zoonotic parasite of high concern. In Canada, an increase in the number of human and animal cases diagnosed has been reported, but information regarding the parasite's distribution in wildlife reservoir remains limited. A cross-sectional study was conducted to estimate the prevalence of wild canids infected with Echinococcus spp. and Echinococcus multilocularis in areas surrounding populated zones in Québec (Canada); to investigate the presence of areas at higher risk of infection; to evaluate potential risk factors of the infection; and as a secondary objective, to compare coproscopy and RT-PCR diagnostic tests for Taenia spp. and Echinococcus identification. From October 2020 to March 2021, fecal samples were collected from 423 coyotes (Canis latrans) and 284 red foxes (Vulpes vulpes) trapped in 12 administrative regions. Real-time PCR for molecular detection of genus Echinococcus spp. and species-specific Echinococcus multilocularis were performed. A total of 38 positive cases of Echinococcus spp., of which 25 were identified as E. multilocularis, were detected. Two high-risk areas of infection were identified. The prevalence of Echinococcus spp. was 22.7% (95% CI 11.5-37.8%) in the Montérégie centered high-risk area, 26.5% (95% CI 12.9-44.4%) in the Bas-St-Laurent high-risk area, and 3.0% (95%CI 1.8-4.7%) outside those areas. For E. multilocularis, a prevalence of 20.5% (95% CI 9.8-35.3%) was estimated in the high-risk area centered in Montérégie compared to 2.4% (95% CI 1.4-3.9%) outside. Logistic regression did not show any association of infection status with species, sex, or geolocation of capture (p > 0.05). This study shows the circulation of Echinococcus in a wildlife cycle in 9/12 administrative regions of Québec.


Asunto(s)
Animales Salvajes , Equinococosis , Echinococcus , Zorros , Animales , Quebec/epidemiología , Equinococosis/epidemiología , Equinococosis/veterinaria , Equinococosis/parasitología , Prevalencia , Animales Salvajes/parasitología , Echinococcus/genética , Echinococcus/aislamiento & purificación , Estudios Transversales , Zorros/parasitología , Echinococcus multilocularis/aislamiento & purificación , Echinococcus multilocularis/genética , Heces/parasitología , Canidae/parasitología , Coyotes/parasitología
3.
Med Vet Entomol ; 2024 Jul 16.
Artículo en Inglés | MEDLINE | ID: mdl-39011830

RESUMEN

Leishmania spp. are zoonotic parasites transmitted by phlebotomine sand flies, including those of the Lutzomyia genus, which can cause leishmaniases in both humans and dogs. Lutzomyia spp. are established in many countries in South and Central America and some areas of the southern United States, with suspected potential of these vectors to undergo further range expansion due to climate change. A scoping review was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extensions for Scoping Reviews (PRISMA-ScR) guidelines to describe the current state of knowledge on the key ecological factors associated with Lutzomyia spp. survival, reproduction and establishment. The following electronic databases were searched for eligible studies published from 1 January 1990, to the date of search, 26 April 2023: CAB Direct (CABI), MEDLINE (via Ovid), Biological Sciences Database and Environmental Sciences Database. Primary research articles that were available in English and focused on ecological factors associated with Lutzomyia spp., such as climatic and habitat factors, geographic range, seasonality and temporality, and host abundance, were eligible for inclusion in the study. Following de-duplication, a total of 167 studies were included in Level 1 screening, 64 studies were included in Level 2 screening and 31 studies met the criteria for data extraction. Study locations included Argentina, Brazil, Colombia, Peru, Venezuela, the United States, Mexico and Canada, with some studies including multiple regions. A total of 31 different Lutzomyia spp. were assessed across these studies, with most (51.6%) of the studies focused on Lutzomyia longipalpis. Eligible studies investigated factors such as seasonality (n = 5), temperature (n = 19), precipitation (n = 13), humidity (n = 2), vegetation presence or requirements (n = 13), ecotypes (n = 7), and/or community type (i.e., urban, suburban, rural) (n = 5). Lutzomyia spp. activity was found to be higher during the rainy season, and peak when temperatures were between 20 and 25°C. Lutzomyia spp. were also found to preferentially reside in tropical or subtropical forests, which are characterised by their lack of a distinct dry season and high precipitation. This scoping review summarised the current state of the literature on the ecological factors associated with the survival, activity and reproduction of Lutzomyia spp. While there appears to be some consensus in the literature regarding some ecological requirements (such as seasonality, temperature and habitat features), overall, there is a lack of published research in this topic. This poses a significant challenge for future studies, which aim to predict the future distribution of Lutzomyia spp. in the context of climate and land use changes. Additional ecological research is urgently needed on Lutzomyia spp. given their relevance to both human and animal health.

4.
mBio ; 15(7): e0047724, 2024 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-38864609

RESUMEN

Parasites of the genus Leishmania pose a global health threat with limited treatment options. New drugs are urgently needed, and genomic screens have the potential to accelerate target discovery, mode of action, and resistance mechanisms against these new drugs. We describe here our effort in developing a genome-wide CRISPR-Cas9 screen in Leishmania, an organism lacking a functional nonhomologous end joining system that must rely on microhomology-mediated end joining, single-strand annealing, or homologous recombination for repairing Cas9-induced double-stranded DNA breaks. A new vector for cloning and expressing single guide RNAs (sgRNAs) was designed and proven to be effective in a small pilot project while enriching specific sgRNAs during drug selection. We then developed a whole-genome library of 49,754 sgRNAs, targeting all the genes of Leishmania infantum. This library was transfected in L. infantum expressing Cas9, and these cells were selected for resistance to two antileishmanials, miltefosine and amphotericin B. The sgRNAs the most enriched in the miltefosine screen targeted the miltefosine transporter gene, but sgRNAs targeting genes coding for a RING-variant protein and a transmembrane protein were also enriched. The sgRNAs the most enriched by amphotericin B targeted the sterol 24 C methyltransferase genes and a hypothetical gene. Through gene disruption experiments, we proved that loss of function of these genes was associated with resistance. This study describes the feasibility of carrying out whole-genome CRISPR-Cas9 screens in Leishmania provided that a strong selective pressure is applied. Such a screen can be used for accelerating the development of urgently needed antileishmanial drugs.IMPORTANCELeishmaniasis, a global health threat, lacks adequate treatment options and drug resistance exacerbates the challenge. This study introduces a CRISPR-Cas9 screening approach in Leishmania infantum, unraveling mechanisms of drug resistance at a genome-wide scale. Our screen was applied against two main antileishmanial drugs, and guides were enriched upon drug selection. These guides targeted known and new targets, hence validating the use of this screen against Leishmania. This strategy provides a powerful tool to expedite drug discovery as well as potential therapeutic targets against this neglected tropical disease.


Asunto(s)
Antiprotozoarios , Sistemas CRISPR-Cas , Resistencia a Medicamentos , Ensayos Analíticos de Alto Rendimiento , Leishmania infantum , Leishmania infantum/genética , Leishmania infantum/efectos de los fármacos , Resistencia a Medicamentos/genética , Antiprotozoarios/farmacología , Ensayos Analíticos de Alto Rendimiento/métodos , Fosforilcolina/farmacología , Fosforilcolina/análogos & derivados , Anfotericina B/farmacología , ARN Guía de Sistemas CRISPR-Cas/genética , Genoma de Protozoos
5.
Traffic ; 25(4): e12935, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38629580

RESUMEN

The protozoan parasites Plasmodium falciparum, Leishmania spp. and Trypanosoma cruzi continue to exert a significant toll on the disease landscape of the human population in sub-Saharan Africa and Latin America. Control measures have helped reduce the burden of their respective diseases-malaria, leishmaniasis and Chagas disease-in endemic regions. However, the need for new drugs, innovative vaccination strategies and molecular markers of disease severity and outcomes has emerged because of developing antimicrobial drug resistance, comparatively inadequate or absent vaccines, and a lack of trustworthy markers of morbid outcomes. Extracellular vesicles (EVs) have been widely reported to play a role in the biology and pathogenicity of P. falciparum, Leishmania spp. and T. cruzi ever since they were discovered. EVs are secreted by a yet to be fully understood mechanism in protozoans into the extracellular milieu and carry a cargo of diverse molecules that reflect the originator cell's metabolic state. Although our understanding of the biogenesis and function of EVs continues to deepen, the question of how EVs in P. falciparum, Leishmania spp. and T. cruzi can serve as targets for a translational agenda into clinical and public health interventions is yet to be fully explored. Here, as a consortium of protozoan researchers, we outline a plan for future researchers and pose three questions to direct an EV's translational agenda in P. falciparum, Leishmania spp. and T. cruzi. We opine that in the long term, executing this blueprint will help bridge the current unmet needs of these medically important protozoan diseases in sub-Saharan Africa and Latin America.


Asunto(s)
Enfermedad de Chagas , Vesículas Extracelulares , Leishmania , Parásitos , Trypanosoma cruzi , Animales , Humanos , Enfermedad de Chagas/epidemiología , Enfermedad de Chagas/parasitología
6.
PLoS Negl Trop Dis ; 18(2): e0012015, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38422164

RESUMEN

BACKGROUND: Visceral leishmaniasis (VL) resolution depends on a wide range of factors, including the instauration of an effective treatment coupled to a functional host immune system. Patients with a depressed immune system, like the ones receiving methotrexate (MTX), are at higher risk of developing VL and refusing antileishmanial drugs. Moreover, the alarmingly growing levels of antimicrobial resistance, especially in endemic areas, contribute to the increasing the burden of this complex zoonotic disease. PRINCIPAL FINDINGS: To understand the potential links between immunosuppressants and antileishmanial drugs, we have studied the interaction of antimony (Sb) and MTX in a Leishmania infantum reference strain (LiWT) and in two L. infantum clinical strains (LiFS-A and LiFS-B) naturally circulating in non-treated VL dogs in Spain. The LiFS-A strain was isolated before Sb treatment in a case that responded positively to the treatment, while the LiFS-B strain was recovered from a dog before Sb treatment, with the dog later relapsing after the treatment. Our results show that, exposure to Sb or MTX leads to an increase in the production of reactive oxygen species (ROS) in LiWT which correlates with a sensitive phenotype against both drugs in promastigotes and intracellular amastigotes. LiFS-A was sensitive against Sb but resistant against MTX, displaying high levels of protection against ROS when exposed to MTX. LiFS-B was resistant to both drugs. Evaluation of the melting proteomes of the two LiFS, in the presence and absence of Sb and MTX, showed a differential enrichment of direct and indirect targets for both drugs, including common and unique pathways. CONCLUSION: Our results show the potential selection of Sb-MTX cross-resistant parasites in the field, pointing to the possibility to undermine antileishmanial treatment of those patients being treated with immunosuppressant drugs in Leishmania endemic areas.


Asunto(s)
Antiprotozoarios , Leishmania infantum , Leishmaniasis Visceral , Humanos , Animales , Perros , Metotrexato/farmacología , Metotrexato/uso terapéutico , Antimonio/farmacología , Antimonio/uso terapéutico , Especies Reactivas de Oxígeno , Antiprotozoarios/farmacología , Antiprotozoarios/uso terapéutico , Leishmaniasis Visceral/tratamiento farmacológico , Leishmaniasis Visceral/veterinaria , Resistencia a Medicamentos
7.
Biomedicines ; 11(9)2023 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-37760981

RESUMEN

Assessment of structure-activity relationships for anti-protozoan activity revealed a strategy for preparing potent anisomycin derivatives with reduced host toxicity. Thirteen anisomycin analogs were synthesized by modifying the alcohol, amine, and aromatic functional groups. Examination of anti-protozoal activity against various strains of Leishmania and cytotoxicity against leucocytes with comparison against the parent natural product demonstrated typical losses of activity with modifications of the alcohol, amine, and aromatic meta-positions. On the other hand, the para-phenol moiety of anisomycin proved an effective location for introducing substituents without significant loss of anti-protozoan potency. An entry point for differentiating activity against Leishmania versus host has been uncovered by this systematic study.

8.
Prev Med Rep ; 33: 102210, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-37090822

RESUMEN

The COVID-19 pandemic and containment measures will likely have a detrimental impact on immunosuppressed individuals' lifestyle behaviours. Increasing evidence suggests that pet ownership is positively associated with healthier lifestyle. Yet, no study has investigated the potential benefits of pet ownership on lifestyle behaviours of immunosuppressed individuals, a population at increased risk of COVID-19 complications. This study aims to examine 1) changes in light, moderate and vigorous intensity physical activity (LPA, MPA, VPA), sedentary time (SED), and sleep duration, assessed by comparing "before COVID-19 pandemic" and "past 7 days" (i.e., current, during pandemic) self-reported behaviours in immunosuppressed individuals and their relatives; 2) to assess if changes in lifestyle behaviours are associated with pet ownership status and whether age is a moderator of these associations. A convenience sample of 132 participants (65.2% female, 41.3% ≥55 years of age) provided self-reported LPA, MPA, VPA (days/week), SED and sleep (min/day) and pet ownership status using an online questionnaire (May-August 2020). Descriptive analyses, paired T-tests, Cohen's d effect size and linear regressions were conducted. Results show that participants reported a decrease in VPA (-0.56 days/week, d = 0.34; p < 0.01) and an increase in SED (106.79 min/day, d = -0.81; p < 0.01). Stratified analysis revealed that having at least one dog, compared to not owning pets, is associated with a reduced decline in LPA, MPA and VPA and an increase in sleep in participants aged < 55 years old only. Having a dog appears to be positively associated with healthy lifestyle behaviours in younger and middle age immunosuppressed individuals.

9.
STAR Protoc ; 4(2): 102248, 2023 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-37087735

RESUMEN

Here, we focus on Leishmania extracellular vesicles (EVs) and their DNA content, detailing a protocol for the isolation of these nanoparticles and their subsequent genomic characterization. We describe a robust and comprehensive approach for obtaining, storing, and analyzing EVs derived from cultured parasites. We detail a user-friendly bioinformatics pipeline for sequence analysis and visualization of CNV analysis and ploidy changes. For complete details on the use and execution of this protocol, please refer to Douanne et al. (2022).1.

10.
J Extracell Biol ; 2(10): e117, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-38939734

RESUMEN

Parasites are responsible for the most neglected tropical diseases, affecting over a billion people worldwide (WHO, 2015) and accounting for billions of cases a year and responsible for several millions of deaths. Research on extracellular vesicles (EVs) has increased in recent years and demonstrated that EVs shed by pathogenic parasites interact with host cells playing an important role in the parasite's survival, such as facilitation of infection, immunomodulation, parasite adaptation to the host environment and the transfer of drug resistance factors. Thus, EVs released by parasites mediate parasite-parasite and parasite-host intercellular communication. In addition, they are being explored as biomarkers of asymptomatic infections and disease prognosis after drug treatment. However, most current protocols used for the isolation, size determination, quantification and characterization of molecular cargo of EVs lack greater rigor, standardization, and adequate quality controls to certify the enrichment or purity of the ensuing bioproducts. We are now initiating major guidelines based on the evolution of collective knowledge in recent years. The main points covered in this position paper are methods for the isolation and molecular characterization of EVs obtained from parasite-infected cell cultures, experimental animals, and patients. The guideline also includes a discussion of suggested protocols and functional assays in host cells.

11.
Cells ; 11(21)2022 10 24.
Artículo en Inglés | MEDLINE | ID: mdl-36359743

RESUMEN

Extracellular vesicles (EVs) contribute to intercellular communication through the transfer of their rich cargo to recipient cells. The EVs produced by LPS-stimulated neutrophils from healthy humans and horses increase airway smooth muscle (ASM) proliferation, but the roles of neutrophil EVs in asthma are largely unexplored. The aim of this study was to determine whether neutrophil-derived EVs isolated during the remission or exacerbation of asthma influence ASM proliferation differentially. Peripheral blood neutrophils were collected during remission and exacerbation in eight horses affected by severe asthma. The cells were cultured (±LPS), and their EVs were isolated by ultracentrifugation and characterized by laser scattering microscopy and proteomic analysis. The proliferation of ASM co-incubated with EVs was monitored in real time by electrical impedance. Two proteins were significantly upregulated during disease exacerbation in neutrophil EVs (MAST4 and Lrch4), while LPS stimulation greatly altered the proteomic profile. Those changes involved the upregulation of neutrophil degranulation products, including proteases known to induce myocyte proliferation. In agreement with the proteomic results, EVs from LPS-stimulated neutrophils increased ASM proliferation, without an effect of the disease status. The inhalation of environmental LPS could contribute to asthma pathogenesis by activating neutrophils and leading to ASM hyperplasia.


Asunto(s)
Asma , Vesículas Extracelulares , Humanos , Caballos , Animales , Neutrófilos/metabolismo , Proteómica , Lipopolisacáridos/farmacología , Proliferación Celular , Músculo Liso/metabolismo , Asma/patología , Vesículas Extracelulares/metabolismo , Proteínas Asociadas a Microtúbulos , Proteínas Serina-Treonina Quinasas
13.
Front Cell Infect Microbiol ; 12: 954144, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35992178

RESUMEN

Visceral leishmaniasis (VL), caused by Leishmania infantum, is an oft-fatal neglected tropical disease. In the absence of an effective vaccine, the control of leishmaniasis relies exclusively on chemotherapy. Due to the lack of established molecular/genetic markers denoting parasite resistance, clinical treatment failure is often used as an indicator. Antimony-based drugs have been the standard antileishmanial treatment for more than seven decades, leading to major drug resistance in certain regions. Likewise, drug resistance to miltefosine and amphotericin B continues to spread at alarming rates. In consequence, innovative approaches are needed to accelerate the identification of antimicrobial drug targets and resistance mechanisms. To this end, we have implemented a novel approach based on thermal proteome profiling (TPP) to further characterize the mode of action of antileishmanials antimony, miltefosine and amphotericin B, as well as to better understand the mechanisms of drug resistance deployed by Leishmania. Proteins become more resistant to heat-induced denaturation when complexed with a ligand. In this way, we used multiplexed quantitative mass spectrometry-based proteomics to monitor the melting profile of thousands of expressed soluble proteins in WT, antimony-resistant, miltefosine-resistant, and amphotericin B-resistant L. infantum parasites, in the presence (or absence) of the above-mentioned drugs. Bioinformatics analyses were performed, including data normalization, melting profile fitting, and identification of proteins that underwent changes (fold change > 4) caused by complexation with a drug. With this unique approach, we were able to narrow down the regions of the L. infantum proteome that interact with antimony, miltefosine, and amphotericin B; validating previously-identified and unveiling novel drug targets. Moreover, analyses revealed candidate proteins potentially involved in drug resistance. Interestingly, we detected thermal proximity coaggregation for several proteins belonging to the same metabolic pathway (i.e., tryparedoxin peroxidase and aspartate aminotransferase in proteins exposed to antimony), highlighting the importance of these pathways. Collectively, our results could serve as a jumping-off point for the future development of innovative diagnostic tools for the detection and evaluation of antimicrobial-resistant Leishmania populations, as well as open the door for new on-target therapies.


Asunto(s)
Antiprotozoarios , Leishmania infantum , Anfotericina B/farmacología , Antimonio/metabolismo , Antimonio/farmacología , Antiprotozoarios/metabolismo , Antiprotozoarios/farmacología , Proteoma/análisis , Proteómica
14.
Cell Rep ; 40(3): 111121, 2022 07 19.
Artículo en Inglés | MEDLINE | ID: mdl-35858561

RESUMEN

Leishmania are eukaryotic parasites that have retained the ability to produce extracellular vesicles (EVs) through evolution. To date, it has been unclear if different DNA entities could be associated with Leishmania EVs and whether these could constitute a mechanism of horizontal gene transfer (HGT). Herein, we investigate the DNA content of EVs derived from drug-resistant parasites, as well as the EVs' potential to act as shuttles for DNA transfer. Next-generation sequencing and PCR assays confirm the enrichment of amplicons carrying drug-resistance genes associated with EVs. Transfer assays of drug-resistant EVs highlight a significant impact on the phenotype of recipient parasites induced by the expression of the transferred DNA. Recipient parasites display an enhanced growth and better control of oxidative stress. We provide evidence that eukaryotic EVs function as efficient mediators in HGT, thereby facilitating the transmission of drug-resistance genes and increasing the fitness of cells when encountering stressful environments.


Asunto(s)
Vesículas Extracelulares , Leishmania , Parásitos , Animales , Resistencia a Medicamentos/genética , Eucariontes , Vesículas Extracelulares/metabolismo , Leishmania/genética , Leishmania/metabolismo
15.
J Zoo Wildl Med ; 53(2): 461-469, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35758589

RESUMEN

In this case series, clinical investigations were pursued during a Synhimantus nasuta infection in a lorikeet (Trichoglossus spp.) flock outbreak situation to better describe and document clinical presentations. In 11 lorikeets suspected to be infected with Synhimantus based on at least one abnormal finding on their physical examination (lethargy, feather-damaging behavior on the ventrum, weight loss, pale iris), the presence of five additional parameters was documented: anemia, relative eosinophilia, increased proventricular diameter-to-keel height ratio (PKR), proventricular barium filling defect, and positive fecal occult blood detection test. A total score (X of 9) was calculated by combining all these findings. Synhimantus nasuta infection was confirmed in four of these individuals by modified Wisconsin fecal examination. Suspected cases (n = 7 of 11) presented only with low scores (1-3 of 9), whereas birds with confirmed infections (n = 4 of 11) presented with both low (1-3 of 9, n = 2 of 4) and high (6-7 of 9, n = 2 of 4) total scores. High scores were associated with clinical anemia. Fecal occult blood was present in all confirmed cases and 4 of 7 suspected cases. An enlarged proventriculus was only observed in birds with active shedding (n = 3 of 4). Follow-up evaluations after 6 mon of treatment with ivermectin and selamectin suggested complete recovery with lowered or normalized total scores. In conclusion, during an S. nasuta outbreak, a rapid physical examination helps to identify suspect cases, including individuals requiring immediate medical attention. In the absence of ova shedding, infection cannot be excluded on the basis of scarce clinical findings, but the detection of occult fecal blood and an increased PKR should raise the index of suspicion.


Asunto(s)
Enfermedades de las Aves , Infecciones por Nematodos , Loros , Espirúridos , Animales , Enfermedades de las Aves/diagnóstico , Enfermedades de las Aves/epidemiología , Infecciones por Nematodos/veterinaria , Proventrículo
17.
Trends Parasitol ; 38(4): 274-276, 2022 04.
Artículo en Inglés | MEDLINE | ID: mdl-35181250

RESUMEN

The selection of Leishmania hybrids in axenic culture was considered rare until recently, when Louradour and Ferreira et al., demonstrated that induced DNA damage facilitates genetic exchange, resulting in full genome tetraploid progenies in vitro. Meiosis-related gene homologues HAP2, GEX1, and RAD51 were found to be involved, opening new avenues for functional genomic studies.


Asunto(s)
Leishmania , Genoma , Hibridación Genética , Leishmania/genética
18.
Parasit Vectors ; 15(1): 5, 2022 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-34983616

RESUMEN

BACKGROUND: Asymptomatic Leishmania infection may play an important role in the transmission of the parasite in endemic areas. At present there is no consensus on the definition of asymptomatic Leishmania infection, nor is there a safe and accessible gold standard test for its identification. METHODS: This paper presents a scoping review to summarize definitions of asymptomatic Leishmania infection found in the literature, as well as to detail the approach (molecular, serological, cellular, and/or parasitological tests) used by researchers to identify this asymptomatic population. A scoping review of published and gray literature related to asymptomatic Leishmania infection was conducted; retrieved citations were screened based on predefined eligibility criteria, and relevant data items were extracted from eligible articles. The analysis is descriptive and is presented using tables, figures, and thematic narrative synthesis. RESULTS: We conducted a screening of 3008 articles, of which 175 were selected for the full review. Of these articles, we selected 106 that met the inclusion criteria. These articles were published between 1991 and 2021, and in the last 5 years, up to 38 articles were reported. Most of the studies were conducted in Brazil (26%), Spain (14%), India (12%), Bangladesh (10%), and Ethiopia (7%). Of the studies, 84.9% were conducted in the immunocompetent population, while 15.1% were conducted in the immunosuppressed population (HIV, immunosuppressive drugs, and organ transplantation population). We report 14 different techniques and 10 strategies employed by researchers to define asymptomatic Leishmania infection in an endemic area. CONCLUSIONS: The definition of asymptomatic Leishmania infection is not unified across the literature, but often includes the following criteria: residence (or extended stay) in a Leishmania-endemic area, no reported signs/symptoms compatible with leishmaniasis, and positive on a combination of serological, molecular, cellular, and/or parasitological tests. Caution is recommended when comparing results of different studies on the subject of asymptomatic infections, as the reported prevalence cannot be confidently compared between areas due to the wide variety of tests employed by research groups. More research on the importance of asymptomatic immunosuppressed and immunocompetent Leishmania-positive populations in leishmaniasis epidemiology is required.


Asunto(s)
Infecciones Asintomáticas , Leishmaniasis/diagnóstico , Enfermedades Endémicas , Humanos , Enfermedades Desatendidas/diagnóstico
19.
Front Vet Sci ; 8: 696815, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34336980

RESUMEN

Cats that spend time outdoors and dogs are particularly at risk of exposure to ticks and the pathogens they transmit. A retrospective study on data collected through passive tick surveillance was conducted to estimate the risk of exposure to tick-borne pathogens in cats and dogs bitten by blacklegged ticks (Ixodes scapularis) in the province of Quebec, Canada, from 2010 to 2017. Blacklegged ticks collected from these host animals were tested by PCR for Borrelia burgdorferi sensu stricto, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti. A total of 13,733 blacklegged ticks were collected from 12,547 animals. Most ticks were adult females and partially engorged. In total, 1,774 cats were infested with ticks and 22.6 and 2.7% of these animals were bitten by at least one tick infected with B. burgdorferi and A. phagocytophilum, respectively. For the 10,773 tick infested dogs, 18.4% were exposed to B. burgdorferi positive ticks while 1.9% of infested dogs were exposed to ticks infected with A. phagocytophilum. The risk of exposure of both cats and dogs to B. miyamotoi and B. microti was lower since only 1.2 and 0.1% of ticks removed were infected with these pathogens, respectively. Traveling outside of the province of Quebec prior to tick collection was significantly associated with exposure to at least one positive tick for B. burgdorferi, A. phagocytophilum and B. microti. Animals exposed to B. burgdorferi or B. miyamotoi positive tick(s) were at higher risk of being concurrently exposed to A. phagocytophilum; higher risk of exposure to B. microti was also observed in animals concurrently exposed to B. burgdorferi. The odds of dogs having B. burgdorferi antibodies were higher when multiple ticks were collected on an animal. The testing and treatment strategies used on dogs bitten by infected ticks were diverse, and misconceptions among veterinarians regarding the treatment of asymptomatic but B. burgdorferi-seropositive dogs were noted. In conclusion, our study demonstrates that cats and dogs throughout Quebec are exposed to blacklegged ticks infected with B. burgdorferi and A. phagocytophilum, and veterinarians across the province need to be aware of this potential threat to the health of pets and their owners.

20.
Parasit Vectors ; 14(1): 438, 2021 Aug 28.
Artículo en Inglés | MEDLINE | ID: mdl-34454601

RESUMEN

BACKGROUND: The evolution of drug resistance is one of the biggest challenges in leishmaniasis and has prompted the need for new antileishmanial drugs. Repurposing of approved drugs is a faster and very attractive strategy that is gaining supporters worldwide. Different anticancer topoisomerase 1B (TOP1B) inhibitors have shown strong antileishmanial activity and promising selective indices, supporting the potential repurposing of these drugs. However, cancer cells and Leishmania share the ability to become rapidly resistant. The aim of this study was to complete a whole-genome exploration of the effects caused by exposure to topotecan in order to highlight the potential mechanisms deployed by Leishmania to favor its survival in the presence of a TOP1B inhibitor. METHODS: We used a combination of stepwise drug resistance selection, whole-genome sequencing, functional validation, and theoretical approaches to explore the propensity of and potential mechanisms deployed by three independent clones of L. infantum to resist the action of TOP1B inhibitor topotecan. RESULTS: We demonstrated that L. infantum is capable of becoming resistant to high concentrations of topotecan without impaired growth ability. No gene deletions or amplifications were identified from the next-generation sequencing data in any of the three resistant lines, ruling out the overexpression of efflux pumps as the preferred mechanism of topotecan resistance. We identified three different mutations in the large subunit of the leishmanial TOP1B (Top1BF187Y, Top1BG191A, and Top1BW232R). Overexpression of these mutated alleles in the wild-type background led to high levels of resistance to topotecan. Computational molecular dynamics simulations, in both covalent and non-covalent complexes, showed that these mutations have an effect on the arrangement of the catalytic pentad and on the interaction of these residues with surrounding amino acids and DNA. This altered architecture of the binding pocket results in decreased persistence of topotecan in the ternary complex. CONCLUSIONS: This work helps elucidate the previously unclear potential mechanisms of topotecan resistance in Leishmania by mutations in the large subunit of TOP1B and provides a valuable clue for the design of improved inhibitors to combat resistance in both leishmaniasis and cancer. Our data highlights the importance of including drug resistance evaluation in drug discovery cascades.


Asunto(s)
Antiprotozoarios/farmacología , ADN-Topoisomerasas de Tipo I/genética , Resistencia a Medicamentos , Leishmania infantum/efectos de los fármacos , Leishmania infantum/genética , Mutación , Inhibidores de Topoisomerasa I/farmacología , Topotecan/farmacología , Antineoplásicos/farmacología , Reposicionamiento de Medicamentos , Leishmania infantum/enzimología , Leishmaniasis/parasitología , Simulación de Dinámica Molecular , Secuenciación Completa del Genoma
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