scholarly journals Recent Advances in Molecular Genetic Tools for Babesia

2021 ◽  
Vol 8 (10) ◽  
pp. 222
Author(s):  
Hassan Hakimi ◽  
Masahito Asada ◽  
Shin-ichiro Kawazu
Keyword(s):  
Drug Targets ◽  
Molecular Genetic ◽  
Essential Genes ◽  
Babesia Bovis ◽  
Bovine Babesiosis ◽  
Genetic Tools ◽  
Genome Wide ◽  
Glms Ribozyme ◽  
Novel Drug

Development of in vitro culture and completion of genome sequencing of several Babesia parasites promoted the efforts to establish transfection systems for these parasites to dissect the gene functions. It has been more than a decade since the establishment of first transfection for Babesia bovis, the causative agent of bovine babesiosis. However, the number of genes that were targeted by genetic tools in Babesia parasites is limited. This is partially due to the low efficiencies of these methods. The recent adaptation of CRISPR/Cas9 for genome editing of Babesia bovis can accelerate the efforts for dissecting this parasite’s genome and extend the knowledge on biological aspects of erythrocytic and tick stages of Babesia. Additionally, glmS ribozyme as a conditional knockdown system is available that could be used for the characterization of essential genes. The development of high throughput genetic tools is needed to dissect the function of multigene families, targeting several genes in a specific pathway, and finally genome-wide identification of essential genes to find novel drug targets. In this review, we summarized the current tools that are available for Babesia and the genes that are being targeted by these tools. This may draw a perspective for the future development of genetic tools and pave the way for the identification of novel drugs or vaccine targets.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Author(s):  
Carlos Suarez ◽  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
Apicomplexan Parasites ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of actively replicating horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by ELISA. Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 hr among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQs-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 hours. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 have a significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

scholarly journals Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 143 ◽  
Author(s):  
J. Antonio Alvarez ◽  
Carmen Rojas ◽  
Julio V. Figueroa
Keyword(s):  
Clinical Signs ◽  
International Health ◽  
Babesia Bovis ◽  
Diagnostic Tools ◽  
Infected Cattle ◽  
Babesia Bigemina ◽  
Bovine Babesiosis ◽  
Culture Methods ◽  
Persistently Infected

Bovine babesiosis is a tick-borne disease of cattle caused by the protozoan parasites of the genus Babesia. Babesia bovis, Babesia bigemina and Babesia divergens are considered by International health authorities (OIE) as the principal species of Babesia that cause bovine babesiosis. Animals that recover from a babesial primo infection may remain as persistent carriers with no clinical signs of disease and can be the source of infection for ticks that are able to acquire Babesia parasites from infected cattle and to transmit Babesia parasites to susceptible cattle. Several procedures that have been developed for parasite detection and diagnosis of this infectious carrier state constitute the basis for this review: A brief description of the direct microscopic detection of Babesia-infected erytrocytes; PCR-based diagnostic assays, which are very sensitive particularly in detecting Babesia in carrier cattle; in-vitro culture methods, used to demonstrate presence of carrier infections of Babesia sp.; animal inoculation, particularly for B. divergens isolation are discussed. Alternatively, persistently infected animals can be tested for specific antibabesial antibodies by using indirect serological assays. Serological procedures are not necessarily consistent in identifying persistently infected animals and have the disadvantage of presenting with cross reactions between antibodies to Babesia sp.

Innovative Methodology in the Discovery of Novel Drug Targets in the Free-Living Amoebae

2018 ◽  
Vol 20 (1) ◽  
pp. 60-69 ◽  
Author(s):  
Abdul Mannan Baig
Keyword(s):  
Drug Targets ◽  
Mortality Rates ◽  
In Vitro Assays ◽  
Cancer Drug ◽  
Cns Infection ◽  
Free Living ◽  
Drug Target Discovery ◽  
Novel Drug ◽  
Novel Drug Targets

Despite advances in drug discovery and modifications in the chemotherapeutic regimens, human infections caused by free-living amoebae (FLA) have high mortality rates (~95%). The FLA that cause fatal human cerebral infections include Naegleria fowleri, Balamuthia mandrillaris and Acanthamoeba spp. Novel drug-target discovery remains the only viable option to tackle these central nervous system (CNS) infection in order to lower the mortality rates caused by the FLA. Of these FLA, N. fowleri causes primary amoebic meningoencephalitis (PAM), while the A. castellanii and B. Mandrillaris are known to cause granulomatous amoebic encephalitis (GAE). The infections caused by the FLA have been treated with drugs like Rifampin, Fluconazole, Amphotericin-B and Miltefosine. Miltefosine is an anti-leishmanial agent and an experimental anti-cancer drug. With only rare incidences of success, these drugs have remained unsuccessful to lower the mortality rates of the cerebral infection caused by FLA. Recently, with the help of bioinformatic computational tools and the discovered genomic data of the FLA, discovery of newer drug targets has become possible. These cellular targets are proteins that are either unique to the FLA or shared between the humans and these unicellular eukaryotes. The latter group of proteins has shown to be targets of some FDA approved drugs prescribed in non-infectious diseases. This review out-lines the bioinformatics methodologies that can be used in the discovery of such novel drug-targets, their chronicle by in-vitro assays done in the past and the translational value of such target discoveries in human diseases caused by FLA.

scholarly journals Development of a Tightly Regulatable Copper-Mediated Gene Switch System in Dermatophytes

2012 ◽  
Vol 78 (15) ◽  
pp. 5204-5211 ◽  
Author(s):  
Atsushi Iwata ◽  
Mohamed Mahdi Alshahni ◽  
Yayoi Nishiyama ◽  
Koichi Makimura ◽  
Shigeru Abe ◽  
...  
Keyword(s):  
Drug Targets ◽  
Efflux Pump ◽  
Molecular Genetic ◽  
Genetic Research ◽  
Promoter Sequence ◽  
Essential Genes ◽  
Regulation System ◽  
Copper Transporter ◽  
Content Type ◽  
Targeted Gene Deletion

ABSTRACTTargeted gene deletion is now available for molecular genetic research of dermatophytes, and the physiological roles of several genes have been elucidated. However, this method cannot be applied to essential genes, which can be potential drug targets. To overcome this limitation, we have developed a conditional gene knockdown system using a copper-responsive promoter. The promoter sequence of the copper transporter geneCTR4(PCTR4) and that of the copper efflux pump geneCRP1(PCRP1) derived fromTrichophyton rubrumwere examined for their response to copper inArthroderma vanbreuseghemii. PCTR4was demonstrated to repress expression of a reporter gene in the presence of copper, while the activity of PCRP1was induced by addition of copper. Importantly, PCTR4regulated the gene expression more tightly. Furthermore, when PCTR4was applied to regulate the expression of the endogenous genesERG1andTRP5, their conditional mutants exhibited decreased growth activity under the repressive conditions. These results suggest that the PCTR4-based gene regulation system represents a powerful tool for identification and characterization of a broad range of genes, including essential genes, in dermatophytes.

scholarly journals Endochin-like quinolone-300 and ELQ-316 inhibit Babesia bovis, B. bigemina, B. caballi and Theileria equi

2020 ◽  
Author(s):  
Marta G. Silva ◽  
Reginaldo G. Bastos ◽  
J. Stone Doggett ◽  
Michael K. Riscoe ◽  
Sovitj Pou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Colorimetric Assay ◽  
Babesia Bovis ◽  
Host Cells ◽  
Bovine Babesiosis ◽  
Theileria Equi ◽  
Peripheral Blood Mononuclear ◽  
Equine Piroplasmosis ◽  
And Control

Abstract Background: The most common apicomplexan parasites causing bovine babesiosis are Babesia bovis and B. bigemina, while B. caballi and Theileria equi are responsible for equine piroplasmosis. Treatment and control of these diseases are usually achieved using potentially toxic chemotherapeutics, such as imidocarb diproprionate, but drug-resistant parasites are emerging, and alternative effective and safer drugs are needed. Endochin-like quinolones (ELQ)-300 and ELQ-316 proved safe and efficacious against related apicomplexans, such as Plasmodium spp., and ELQ-316 was also effective against B. microti, without showing toxicity in mammals.Methods: Inhibitory effects of ELQ-300 and ELQ-316 were assessed on the growth of cultured B. bovis, B. bigemina, B. caballi and T. equi. Percentage of parasitized erythrocytes was measured by flow cytometry. Effect of the ELQ drugs on the viability of horse and bovine peripheral blood mononuclear cells (PBMC) was assessed by monitoring cell metabolic activity using a colorimetric assay.Results: We calculated IC50 ranging from 0.04 to 0.37 nM for ELQ-300, and from 0.002 to 0.1 nM for ELQ-316 at 72 h among all cultured parasites tested. None of the parasites tested were able to replicate in cultures in the presence of the ELQ-300 and ELQ-316 at IC100, which range from 1.3 to 5.7 nM for ELQ-300 and from 1.0 to 6.0 nM for ELQ-316 at 72 h. Neither ELQ-300 nor ELQ-316 altered the viability of equine and bovine PBMC at their IC100 in in vitro testing. Conclusions: ELQ-300 and ELQ-316 showed significant inhibitory activity on the main parasites responsible for bovine babesiosis and equine piroplasmosis at doses that are tolerable to host cells. These ELQ drugs may be viable candidates for developing alternative protocols for the treatment of bovine babesiosis and equine piroplasmosis.

Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium

2018 ◽  
Vol 67 (2) ◽  
pp. 190-195 ◽  
Author(s):  
Carmen Rojas-Martínez ◽  
Roger Iván Rodríguez-Vivas ◽  
Julio Vicente Figueroa Millán ◽  
Carlos Ramón Bautista-Garfias ◽  
Roberto Omar Castañeda-Arriola ◽  
...  
Keyword(s):  
Babesia Bovis ◽  
Serum Free Medium ◽  
Free Medium ◽  
Babesia Bigemina ◽  
Bovine Babesiosis ◽  
Serum Free

scholarly journals Identifying nootropic drug targets via large-scale cognitive GWAS and transcriptomics

2021 ◽  
Author(s):  
Max Lam ◽  
Chia-Yen Chen ◽  
Tian Ge ◽  
Yan Xia ◽  
David W. Hill ◽  
...  
Keyword(s):  
Drug Targets ◽  
Large Scale ◽  
Association Studies ◽  
Drug Repurposing ◽  
Genome Wide Association Studies ◽  
Data Set ◽  
Nootropic Drug ◽  
Gene Sets ◽  
Genome Wide ◽  
Novel Drug

AbstractBroad-based cognitive deficits are an enduring and disabling symptom for many patients with severe mental illness, and these impairments are inadequately addressed by current medications. While novel drug targets for schizophrenia and depression have emerged from recent large-scale genome-wide association studies (GWAS) of these psychiatric disorders, GWAS of general cognitive ability can suggest potential targets for nootropic drug repurposing. Here, we (1) meta-analyze results from two recent cognitive GWAS to further enhance power for locus discovery; (2) employ several complementary transcriptomic methods to identify genes in these loci that are credibly associated with cognition; and (3) further annotate the resulting genes using multiple chemoinformatic databases to identify “druggable” targets. Using our meta-analytic data set (N = 373,617), we identified 241 independent cognition-associated loci (29 novel), and 76 genes were identified by 2 or more methods of gene identification. Actin and chromatin binding gene sets were identified as novel pathways that could be targeted via drug repurposing. Leveraging our transcriptomic and chemoinformatic databases, we identified 16 putative genes targeted by existing drugs potentially available for cognitive repurposing.

scholarly journals Pathway analysis of GWAS loci identifies novel drug targets and repurposing opportunities

2018 ◽  
Author(s):  
Deepali Jhamb ◽  
Michal Magid-Slav ◽  
Mark R. Hurle ◽  
Pankaj Agarwal
Keyword(s):  
Drug Discovery ◽  
Pathway Analysis ◽  
Drug Targets ◽  
Global Analysis ◽  
Association Studies ◽  
Genome Wide Association Studies ◽  
Genome Wide ◽  
If Current ◽  
Novel Drug ◽  
Novel Drug Targets

AbstractGenome-wide association studies (GWAS) have made considerable progress and there is emerging evidence that genetics-based targets can lead to 28% more launched drugs. However, translating the results of GWAS for drug discovery remains challenging. We analyzed 1,589 GWAS across 1,456 protein interaction pathways to translate these often, imprecise genetic loci into therapeutic hypotheses for 182 diseases. We validate these pathway-based genetic targets by testing if current drug targets are enriched in the pathway space of the same indication. Remarkably, 30% of diseases have significantly more targets in these pathways than expected by chance; the comparable number for GWAS alone (without using pathway analysis) is zero. Although pathway analysis is routine for GWAS, this study shows that the routine analysis can often enrich for drug targets, by performing a systematic global analysis to translate genetic findings into therapeutic hypotheses for new drug discovery and repositioning opportunities for current drugs.

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