20 results on '"Mehdi Labaied"'
Search Results
2. Structures of plasmepsin X from P. falciparum reveal a novel inactivation mechanism of the zymogen and molecular basis for binding of inhibitors in mature enzyme
- Author
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Alla Gustchina, Pooja Kesari, Vandana Mishra, Mehdi Labaied, Huogen Xiao, Nikhil Pahelkar, Thomas E. Edwards, Anuradha Deshmukh, John H. Dupuis, Alexander Wlodawer, Prasenjit Bhaumik, Abhishek B. Suryawanshi, Ishan Rathore, Rickey Y. Yada, Donald D. Lorimer, and Jan Abendroth
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chemistry.chemical_classification ,Proteases ,Enzyme ,Biochemistry ,Arginine ,biology ,Chemistry ,Zymogen ,Plasmepsin ,Substrate (chemistry) ,Plasmodium falciparum ,Cleavage (embryo) ,biology.organism_classification - Abstract
Plasmodium falciparum plasmepsin X (PfPMX), involved in the invasion and egress of this deadliest malarial parasite, is essential for its survival and hence considered as an important drug target. We report the first crystal structure of PfPMX zymogen containing a novel fold of its prosegment. A unique twisted loop from the prosegment and arginine 244 from the mature enzyme are involved in zymogen inactivation; such mechanism, not previously reported, might be common for apicomplexan proteases similar to PfPMX. The maturation of PfPMX zymogen occurs through cleavage of its prosegment at multiple sites. Our data provide thorough insights into the mode of binding of a substrate and a potent inhibitor 49c to PfPMX. We present molecular details of inactivation, maturation, and inhibition of PfPMX that should aid in the development of potent inhibitors against pepsin-like aspartic proteases from apicomplexan parasites.
- Published
- 2021
3. Computer-Aided Discovery ofTrypanosoma bruceiRNA-Editing Terminal Uridylyl Transferase 2 Inhibitors
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Mehdi Labaied, Özlem Demir, Christopher Merritt, Kenneth Stuart, and Rommie E. Amaro
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Trypanosoma brucei brucei ,Computational biology ,Molecular Dynamics Simulation ,Trypanosoma brucei ,Biochemistry ,Article ,parasitic diseases ,Drug Discovery ,medicine ,Humans ,Parasite hosting ,Transferase ,African trypanosomiasis ,Enzyme Inhibitors ,Pharmacology ,chemistry.chemical_classification ,Virtual screening ,biology ,Drug discovery ,Organic Chemistry ,RNA Nucleotidyltransferases ,biology.organism_classification ,medicine.disease ,Trypanocidal Agents ,Virology ,Molecular Docking Simulation ,Trypanosomiasis, African ,Enzyme ,chemistry ,RNA editing ,Computer-Aided Design ,Molecular Medicine - Abstract
Human African trypanosomiasis (HAT) is a major health problem in sub-Saharan Africa caused by Trypanosoma brucei infection. Current HAT drugs are difficult to administer and not effective against all parasite species at different stages of the disease which indicates an unmet pharmaceutical need. TbRET2 is an indispensable enzyme for the parasite, and is targeted here using a computational approach that combines molecular dynamics simulations and virtual screening. The compounds prioritized are then tested in T. brucei via Alamar blue cell viability assays. This work identified 20 drug-like compounds which are candidates for further testing in the drug discovery process.
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- 2014
4. N1,N1-Dimethyl-N3-(3-(trifluoromethyl)phenethyl)propane-1,3-diamine, a new lead for the treatment of human African trypanosomiasis
- Author
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Kenneth Stuart, Séverine Monnerat, Ronald J. Quinn, Ngoc B. Pham, Sophie M. Deydier, and Mehdi Labaied
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Pharmacology ,Trifluoromethyl ,Natural product ,biology ,Stereochemistry ,Human African trypanosomiasis ,Organic Chemistry ,General Medicine ,Nuclear magnetic resonance spectroscopy ,Trypanosoma brucei ,medicine.disease ,biology.organism_classification ,Anti-protozoa ,chemistry.chemical_compound ,Alkaloids ,chemistry ,Diamine ,parasitic diseases ,Drug Discovery ,medicine ,Potency ,African trypanosomiasis ,Anti-parasite ,Trypanocidal agent - Abstract
The natural product, convolutamine I (1), has anti-trypanosomal activity however it has a high molecular weight of 473 due to a presence of 3 bromine atoms. The synthesis of the natural product convolutamine I (1) together with its analogues are presented. A SAR study against Trypanosoma brucei brucei led to compounds with improved physico-chemical properties: lower molecular weight and lower log P while maintaining potency (with a slight 2-fold improvement).
- Published
- 2014
5. Plasmodium salvages cholesterol internalized by LDL and synthesized de novo in the liver
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Guimin Guan, Mehdi Labaied, Juan Sandoval, Nazneen Bano, Sung Jae Cha, Isabelle Coppens, and Bamini Jayabalasingham
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chemistry.chemical_classification ,Infectivity ,Farnesyl-diphosphate farnesyltransferase ,biology ,Cholesterol ,Immunology ,biology.organism_classification ,Microbiology ,Plasmodium ,chemistry.chemical_compound ,Squalene ,Enzyme ,Biochemistry ,chemistry ,Virology ,parasitic diseases ,lipids (amino acids, peptides, and proteins) ,Receptor ,Lipoprotein - Abstract
Our previous morphological studies illustrated the association of sterols with Plasmodium infecting hepatocytes. Because malaria parasites cannot synthesize sterols, they must scavenge these lipids from the host. In this paper, we have examined the source/s of sterols for intrahepatic Plasmodium and evaluated the importance of sterols for liver stage development. We show that Plasmodium continuously diverts cholesterol from hepatocytes until release of merozoites. Removal of plasma lipoproteins from the medium results in a 70% reduction of cholesterol content in hepatic merozoites but these parasites remain infectious in animals. Plasmodium salvages cholesterol that has been internalized by low-density lipoprotein but reduced expression of host low-density lipoprotein receptors by 70% does not influence liver stage burden. Plasmodium is also able to intercept cholesterol synthesized by hepatocytes. Pharmacological blockade of host squalene synthase or downregulation of the expression of this enzyme by 80% decreases by twofold the cholesterol content of merozoites without further impacting parasite development. These data enlighten that, on one hand, malaria parasites have moderate need of sterols for optimal development in hepatocytes and, on the other hand, they can adapt to survive in cholesterol-restrictive conditions by exploitation of accessible sterols derived from alternative sources in hepatocytes to maintain proper infectivity.
- Published
- 2010
6. Targeted deletion of SAP1 abolishes the expression of infectivity factors necessary for successful malaria parasite liver infection
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Mehdi Labaied, Sebastian A. Mikolajczak, Stefan H. I. Kappe, Vanessa Jacobs-Lorena, Ahmed S. I. Aly, Isabelle Coppens, Nelly Camargo, and Hilda Silva Rivera
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Male ,Protozoan Proteins ,Gene Expression ,Microbiology ,Plasmodium ,Mice ,Cell Line, Tumor ,Gene expression ,parasitic diseases ,Anopheles ,Animals ,Humans ,Molecular Biology ,Gene ,Research Articles ,Sequence Deletion ,Infectivity ,Mice, Inbred BALB C ,Liver infection ,biology ,Effector ,Liver Diseases ,Plasmodium yoelii ,Aly A. S. I. , Mikolajczak S. A. , Rivera H. S. , Camargo N., Jacobs-Lorena V., Labaied M., Coppens I., Kappe S. H. I. , -Targeted deletion of SAP1 abolishes the expression of infectivity factors necessary for successful malaria parasite liver infection-, MOLECULAR MICROBIOLOGY, cilt.69, ss.152-163, 2008 ,biology.organism_classification ,Virology ,Malaria ,Protein Structure, Tertiary ,Phenotype ,Liver ,Sporozoites ,Gene Targeting ,Female - Abstract
Malaria parasite sporozoites prepare for transmission to a mammalian host by upregulation of UIS (Upregulated in Infectious Sporozoites) genes. A number of UIS gene products are essential for the establishment of the intrahepatocytic niche. However, the factors that regulate the expression of genes involved in gain of infectivity for the liver are unknown. Herein, we show that a conserved Plasmodium sporozoite low-complexity asparagine-rich protein, SAP1 (Sporozoite Asparagine-rich Protein 1), has an essential role in malaria parasite liver infection. Targeted deletion of SAP1 in the rodent malaria parasite Plasmodium yoelii generated mutant parasites that traverse and invade hepatocytes normally but cannot initiate liver-stage development in vitro and in vivo. Moreover, immunizations with Pysap1(-) sporozoites confer long-lasting sterile protection against wild-type sporozoite infection. Strikingly, lack of SAP1 abolished expression of essential UIS genes including UIS3, UIS4 and P52 but not the constitutively expressed genes encoding, among others, sporozoite proteins CSP and TRAP. SAP1 localization to the cell interior but not the nucleus of sporozoites suggests its involvement in a post-transcriptional mechanism of gene expression control. These findings demonstrate that SAP1 is essential for liver infection possibly by functioning as a selective regulator controlling the expression of infectivity-associated parasite effector genes.
- Published
- 2008
7. Plasmodium yoelii Sporozoites with Simultaneous Deletion of P52 and P36 Are Completely Attenuated and Confer Sterile Immunity against Infection
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Mehdi Labaied, Anke Harupa, Isabelle Coppens, Ronald F. Dumpit, Stefan H. I. Kappe, and Sebastian A. Mikolajczak
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Cytoplasm ,Immunology ,Protozoan Proteins ,Host cell nucleoplasm ,Microbiology ,Apicomplexa ,Mice ,Microscopy, Electron, Transmission ,Immunity ,parasitic diseases ,Animals ,Plasmodium berghei ,Rats, Wistar ,Infectivity ,biology ,Plasmodium falciparum ,Plasmodium yoelii ,biology.organism_classification ,Virology ,Malaria ,Rats ,Disease Models, Animal ,Culicidae ,Infectious Diseases ,Microscopy, Fluorescence ,Sporozoites ,Gene Targeting ,Vacuoles ,Host cell cytoplasm ,Hepatocytes ,Female ,Parasitology ,Fungal and Parasitic Infections ,Gene Deletion - Abstract
Malaria infection starts when sporozoites are transmitted to the mammalian host during a mosquito bite. Sporozoites enter the blood circulation, reach the liver, and infect hepatocytes. The formation of a parasitophorous vacuole (PV) establishes their intracellular niche. Recently, two members of the 6-Cys domain protein family, P52 and P36, were each shown to play an important albeit nonessential role in Plasmodium berghei sporozoite infectivity for the rodent host. Here, we generated p52/p36 -deficient Plasmodium yoelii parasites by the simultaneous deletion of both genes using a single genetic manipulation. p52/p36 -deficient parasites exhibited normal progression through the life cycle during blood-stage infection, transmission to mosquitoes, mosquito-stage development, and sporozoite infection of the salivary glands. p52/p36 -deficient sporozoites also showed normal motility and cell traversal activity. However, immunofluorescence analysis and electron microscopic observations revealed that p52/p36 -deficient parasites did not form a PV within hepatocytes in vitro and in vivo. The p52/p36 -deficient parasites localized as free entities in the host cell cytoplasm or the host cell nucleoplasm and did not develop as liver stages. Consequently, they did not cause blood-stage infections even at high sporozoite inoculation doses. Mice immunized with p52/p36 -deficient sporozoites were completely protected against infectious sporozoite challenge. Our results demonstrate for the first time the generation of two-locus gene deletion-attenuated parasites that infect the liver but do not progress to blood-stage infection. The study will critically guide the design of Plasmodium falciparum live attenuated malaria vaccines.
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- 2007
8. Synthesis and antimalarial evaluation of a series of piperazinyl flavones
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Gwenola Auffret, Guy Lewin, François Frappier, Philippe Rasoanaivo, Philippe Grellier, Mehdi Labaied, Molécules de Communication et Adaptation des Micro-organismes (MCAM), and Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS)
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[SDV]Life Sciences [q-bio] ,Clinical Biochemistry ,Flavonoid ,Drug Resistance ,Pharmaceutical Science ,Carboxamide ,01 natural sciences ,Biochemistry ,Chemical synthesis ,Piperazines ,Mice ,chemistry.chemical_compound ,Drug Discovery ,ComputingMilieux_MISCELLANEOUS ,chemistry.chemical_classification ,biology ,Chloroquine ,3. Good health ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Molecular Medicine ,Plasmodium yoelii ,Cell Survival ,Stereochemistry ,medicine.drug_class ,Plasmodium falciparum ,010402 general chemistry ,Flavones ,Antimalarials ,Structure-Activity Relationship ,Cell Line, Tumor ,parasitic diseases ,medicine ,Animals ,Humans ,[CHIM]Chemical Sciences ,Structure–activity relationship ,Phenols ,Molecular Biology ,010405 organic chemistry ,Organic Chemistry ,biology.organism_classification ,Malaria ,0104 chemical sciences ,chemistry ,Indicators and Reagents ,Drug Screening Assays, Antitumor ,Genes, MDR - Abstract
A series of 27 flavonoid derivatives containing a piperazinyl chain have been synthesized and tested for their antiplasmodial activity. Diverse substitution patterns on piperazinyl and flavone moieties were examined and found to affect the activity differently. The most active compounds, which have a 2,3,4-trimethoxybenzylpiperazinyl chain attached to the flavone at the 7-phenol group, showed in vitro activity against chloroquine-sensitive (Thai) and -resistant (FcB1,K1) Plasmodium falciparum strains in the micromolar to submicromolar range. One of them was active when given orally in a Plasmodium yoelii nigeriensis infected mouse model.
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- 2007
9. Genetically modified Plasmodium parasites as a protective experimental malaria vaccine
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Mehdi Labaied, Stefan H. I. Kappe, Ann-Kristin Mueller, and Kai Matuschewski
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Plasmodium berghei ,Vaccines, Attenuated ,Plasmodium ,Apicomplexa ,Mice ,Immune system ,Malaria Vaccines ,parasitic diseases ,medicine ,Animals ,Multidisciplinary ,biology ,Malaria vaccine ,Plasmodium falciparum ,medicine.disease ,biology.organism_classification ,Virology ,Malaria ,Genetically modified organism ,Mice, Inbred C57BL ,Phenotype ,Liver ,Immunization ,Gene Targeting ,Female ,Gene Deletion - Abstract
Malaria is a mosquito-borne disease that is transmitted by inoculation of the Plasmodium parasite sporozoite stage. Sporozoites invade hepatocytes, transform into liver stages, and subsequent liver-stage development ultimately results in release of pathogenic merozoites. Liver stages of the parasite are a prime target for malaria vaccines because they can be completely eliminated by sterilizing immune responses, thereby preventing malarial infection. Using expression profiling, we previously identified genes that are only expressed in the pre-erythrocytic stages of the parasite. Here, we show by reverse genetics that one identified gene, UIS3 (upregulated in infective sporozoites gene 3), is essential for early liver-stage development. uis3-deficient sporozoites infect hepatocytes but are unable to establish blood-stage infections in vivo, and thus do not lead to disease. Immunization with uis3-deficient sporozoites confers complete protection against infectious sporozoite challenge in a rodent malaria model. This protection is sustained and stage specific. Our findings demonstrate that a safe and effective, genetically attenuated whole-organism malaria vaccine is possible.
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- 2004
10. Screening extracts of Madagascan plants in search of antiplasmodial compounds
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Dina L Rakotondramanana, Mehdi Labaied, S. Ratsimamanga-Urverg, Lucile Allorge, Philippe Rasoanaivo, David Ramanitrahasimbola, Philippe Grellier, François Frappier, Lengo Mambu, Herintsoa Rafatro, Armand Rakotozafy, and Baholy Robijaona
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Pharmacology ,biology ,Traditional medicine ,Ethyl acetate ,Plasmodium falciparum ,Fractionation ,Pharmacognosy ,medicine.disease ,biology.organism_classification ,In vitro ,law.invention ,chemistry.chemical_compound ,chemistry ,law ,parasitic diseases ,medicine ,Phytotherapy ,IC50 ,Malaria - Abstract
One hundred and ninety plants, of which 51 are used to treat malaria in traditional medicine, were collected in five different ecosystems of Madagascar for a screening programme devoted to the search of naturally-occurring antimalarial compounds. Thirty-nine plants, of which 12 are used as herbal antimalarials, were found to display in vitro activity against Plasmodium falciparum with a median inhibitory concentration (IC50) lower than 5 microg/ml while 9 had an IC50 ranging from 5 to 7.5 microg/ml. Seventeen of them exhibited cytotoxic effects on murine P388 leukemia cells with an IC50 < 10 microg/ml. The biological activities were mostly located in the ethyl acetate fractions. Bioassay-directed fractionation is underway to isolate the active constituents.
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- 2004
11. Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei
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Nicholas Hasle, Savitha Kalidas, Mehdi Labaied, Margaret A. Phillips, Igor Cestari, Kenneth Stuart, Severine Monnerat, Qiong Li, Marilyn Parsons, and Sandesh Regmi
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Trypanosoma brucei brucei ,Protozoan Proteins ,Trypanosoma brucei ,Microbiology ,Amino Acyl-tRNA Synthetases ,chemistry.chemical_compound ,Cytosol ,RNA interference ,Gene expression ,parasitic diseases ,Drug Discovery ,RNA, Messenger ,RNA, Small Interfering ,Molecular Biology ,Gene ,chemistry.chemical_classification ,Gene knockdown ,biology ,Aminoacyl tRNA synthetase ,Drug discovery ,Cell Cycle ,General Medicine ,Articles ,biology.organism_classification ,Trypanocidal Agents ,Mitochondria ,Enzyme ,chemistry ,Biochemistry ,Gene Expression Regulation ,Gene Knockdown Techniques ,Fatty Alcohols ,Signal Transduction - Abstract
Human African trypanosomiasis (HAT) is an important public health threat in sub-Saharan Africa. Current drugs are unsatisfactory, and new drugs are being sought. Few validated enzyme targets are available to support drug discovery efforts, so our goal was to obtain essentiality data on genes with proven utility as drug targets. Aminoacyl-tRNA synthetases (aaRSs) are known drug targets for bacterial and fungal pathogens and are required for protein synthesis. Here we survey the essentiality of eight Trypanosoma brucei aaRSs by RNA interference (RNAi) gene expression knockdown, covering an enzyme from each major aaRS class: valyl-tRNA synthetase (ValRS) (class Ia), tryptophanyl-tRNA synthetase (TrpRS-1) (class Ib), arginyl-tRNA synthetase (ArgRS) (class Ic), glutamyl-tRNA synthetase (GluRS) (class 1c), threonyl-tRNA synthetase (ThrRS) (class IIa), asparaginyl-tRNA synthetase (AsnRS) (class IIb), and phenylalanyl-tRNA synthetase (α and β) (PheRS) (class IIc). Knockdown of mRNA encoding these enzymes in T. brucei mammalian stage parasites showed that all were essential for parasite growth and survival in vitro . The reduced expression resulted in growth, morphological, cell cycle, and DNA content abnormalities. ThrRS was characterized in greater detail, showing that the purified recombinant enzyme displayed ThrRS activity and that the protein localized to both the cytosol and mitochondrion. Borrelidin, a known inhibitor of ThrRS, was an inhibitor of T. brucei ThrRS and showed antitrypanosomal activity. The data show that aaRSs are essential for T. brucei survival and are likely to be excellent targets for drug discovery efforts.
- Published
- 2014
12. N¹,N¹-Dimethyl-N³-(3-(trifluoromethyl)phenethyl)propane-1,3-diamine, a new lead for the treatment of human African trypanosomiasis
- Author
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Ngoc B, Pham, Sophie, Deydier, Mehdi, Labaied, Séverine, Monnerat, Kenneth, Stuart, and Ronald J, Quinn
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Spectrometry, Mass, Electrospray Ionization ,Magnetic Resonance Spectroscopy ,Trypanosomiasis, African ,parasitic diseases ,Phenethylamines ,Humans ,Diamines ,Trypanocidal Agents ,Article - Abstract
The natural product, convolutamine I (1), has anti-trypanosomal activity however it has a high molecular weight of 473 due to a presence of 3 bromine atoms. The synthesis of the natural product convolutamine I (1) together with its analogues are presented. A SAR study against Trypanosoma brucei brucei led to compounds with improved physico-chemical properties: lower molecular weight and lower log P while maintaining potency (with a slight 2-fold improvement).
- Published
- 2013
13. ChemInform Abstract: Synthesis of Antitrypanosomal 1,2-Dioxane Derivatives Based on a Natural Product Scaffold
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Mehdi Labaied, Ngoc Minh Pham, Harish Holla, Ian D. Jenkins, Ronald J. Quinn, and Kenneth Stuart
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chemistry.chemical_compound ,Scaffold ,Natural product ,Chemistry ,Chemical manipulation ,Molecule ,General Medicine ,Peroxide ,Combinatorial chemistry - Abstract
The new scaffold (IV), based on antitrypanosomal and antimalarial compounds isolated from different Plakortis species, contains a peroxide unit which is stable to chemical manipulation elsewhere in the molecule.
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- 2011
14. Synthesis of antitrypanosomal 1,2-dioxane derivatives based on a natural product scaffold
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Ronald J. Quinn, Mehdi Labaied, Kenneth Stuart, Ngoc Minh Pham, Harish Holla, and Ian D. Jenkins
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Clinical Biochemistry ,Trypanosoma brucei brucei ,Pharmaceutical Science ,Stereoisomerism ,Biochemistry ,Peroxide ,Article ,Dioxanes ,Small Molecule Libraries ,chemistry.chemical_compound ,Biological Factors ,Structure-Activity Relationship ,Parasitic Sensitivity Tests ,Plakortis ,Drug Discovery ,Organic chemistry ,Structure–activity relationship ,Molecule ,Animals ,Molecular Biology ,Ene reaction ,Natural product ,Ozonolysis ,Dose-Response Relationship, Drug ,Molecular Structure ,Organic Chemistry ,Combinatorial chemistry ,Trypanocidal Agents ,chemistry ,Wittig reaction ,Molecular Medicine - Abstract
A short practical synthesis of a new natural product based scaffold (6), based on antitrypanosomal and antimalarial compounds isolated from different Plakortis species is described. The scaffold contains a peroxide unit that is surprisingly stable to chemical manipulation elsewhere in the molecule, enabling it to be elaborated into a small library of derivatives. It is stable to ozonolysis, reductive work-up with dimethylsulfide and the Wittig reaction with stabilized phosphorus ylides. The scaffold along with its Wittig analogues has displayed low to sub-micro molar (0.2-3.3 μM) antitrypanosomal activity.
- Published
- 2011
15. Plasmodium salvages cholesterol internalized by LDL and synthesized de novo in the liver
- Author
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Mehdi, Labaied, Bamini, Jayabalasingham, Nazneen, Bano, Sung-Jae, Cha, Juan, Sandoval, Guimin, Guan, and Isabelle, Coppens
- Subjects
Plasmodium ,Merozoites ,Cell Line ,Lipoproteins, LDL ,Mice, Inbred C57BL ,Mice ,Sterols ,Cholesterol ,Culicidae ,Farnesyl-Diphosphate Farnesyltransferase ,Liver ,Receptors, LDL ,Hepatocytes ,Animals ,Humans ,Female ,RNA Interference - Abstract
Our previous morphological studies illustrated the association of sterols with Plasmodium infecting hepatocytes. Because malaria parasites cannot synthesize sterols, they must scavenge these lipids from the host. In this paper, we have examined the source/s of sterols for intrahepatic Plasmodium and evaluated the importance of sterols for liver stage development. We show that Plasmodium continuously diverts cholesterol from hepatocytes until release of merozoites. Removal of plasma lipoproteins from the medium results in a 70% reduction of cholesterol content in hepatic merozoites but these parasites remain infectious in animals. Plasmodium salvages cholesterol that has been internalized by low-density lipoprotein but reduced expression of host low-density lipoprotein receptors by 70% does not influence liver stage burden. Plasmodium is also able to intercept cholesterol synthesized by hepatocytes. Pharmacological blockade of host squalene synthase or downregulation of the expression of this enzyme by 80% decreases by twofold the cholesterol content of merozoites without further impacting parasite development. These data enlighten that, on one hand, malaria parasites have moderate need of sterols for optimal development in hepatocytes and, on the other hand, they can adapt to survive in cholesterol-restrictive conditions by exploitation of accessible sterols derived from alternative sources in hepatocytes to maintain proper infectivity.
- Published
- 2010
16. A dispensable Plasmodium locus for stable transgene expression
- Author
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Mehdi Labaied, Stefan H. I. Kappe, Ashley M. Vaughan, Vanessa Jacobs-Lorena, and Sebastian A. Mikolajczak
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Recombination, Genetic ,biology ,Transgene ,fungi ,Genes, Protozoan ,Spores, Protozoan ,Wild type ,Locus (genetics) ,Plasmodium yoelii ,biology.organism_classification ,Molecular biology ,Article ,Recombinant Proteins ,Plasmid ,Blood ,Liver ,Gene expression ,parasitic diseases ,Plasmodium berghei ,Parasitology ,Transgenes ,Genetic Engineering ,Molecular Biology ,Gene - Abstract
The ribosomal small subunit locus has been used for transgene expression in the rodent malaria parasites, Plasmodium berghei and Plasmodium yoelii , but this strategy utilizes single crossover integration and is thus prone to reversion by plasmid excision. Targeting of the ribosomal subunit locus may also have a negative effect on oocyst development in the mosquito. In P. berghei , the p230 paralog locus has been used for transgene expression. Here, we show that the P. yoelii S1 locus (sporozoite expressed gene 1) (PY05712) is dispensable and can be used for stable transgene expression throughout the parasite life cycle. P. yoelii s1 − parasites show no defect in blood stage replication, oocyst formation, sporozoite production, or liver stage development when compared to P. yoelii wildtype parasites. Further, we show that a fluorescent transgene can be stably expressed from this site. This demonstrates that the S1 locus can be utilized for stable expression of heterologous genes in rodent malaria parasites.
- Published
- 2010
17. Photochemical inactivation with amotosalen and long-wavelength ultraviolet light of Plasmodium and Babesia in platelet and plasma components
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Kent Dupuis, Lynette Sawyer, Mehdi Labaied, Jorge L. Benach, Lily Lin, Sébastien Charneau, Matthias Steiert, Ryan Alfonso, Horacio Gil, Gloria Monsalve, Philippe Grellier, Molécules de Communication et Adaptation des Micro-organismes (MCAM), and Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Blood Platelets ,Amotosalen ,Erythrocytes ,Photochemistry ,Ultraviolet Rays ,[SDV]Life Sciences [q-bio] ,Plasmodium falciparum ,Immunology ,Blood Component Transfusion ,Blood Donors ,030204 cardiovascular system & hematology ,Babesia microti ,Plasmodium ,Mice ,Plasma ,03 medical and health sciences ,0302 clinical medicine ,Babesiosis ,Furocoumarins ,parasitic diseases ,medicine ,Ultraviolet light ,Animals ,Humans ,Immunology and Allergy ,[CHIM]Chemical Sciences ,Platelet ,030212 general & internal medicine ,Malaria, Falciparum ,ComputingMilieux_MISCELLANEOUS ,Infectivity ,biology ,Hematology ,medicine.disease ,biology.organism_classification ,3. Good health ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Babesia ,Blood Component Removal - Abstract
BACKGROUND: Transfusion-transmitted cases of malaria and babesiosis have been well documented. Current efforts to screen out contaminated blood products result in component wastage due to the lack of specific detection methods while donor deferral does not always guarantee safe blood products. This study evaluated the efficacy of a photochemical treatment (PCT) method with amotosalen and long-wavelength ultraviolet light (UVA) to inactivate these agents in red blood cells (RBCs) contaminating platelet (PLT) and plasma components. STUDY DESIGN AND METHODS:Plasmodium falciparum– and Babesia microti–contaminated RBCs seeded into PLT and plasma components were treated with 150 µmol per L amotosalen and 3 J per cm2 UVA. The viability of both pathogens before and after treatment was measured with infectivity assays. Treatment with 150 µmol per L amotosalen and 1 J per cm2 UVA was used to assess the robustness of the PCT system. RESULTS: No viable B. microti was detected in PLTs or plasma after treatment with 150 mol per L amotosalen and 3 J per cm2 UVA, demonstrating a mean inactivation of greater than 5.3 log in PLTs and greater than 5.3 log in plasma. After the same treatment, viable P. falciparum was either absent or below the limit of quantification in three of four replicate experiments both in PLTs and in plasma demonstrating a mean inactivation of at least 6.0 log in PLTs and at least 6.9 log in plasma. Reducing UVA dose to 1 J per cm2 did not significantly affect the level of inactivation. CONCLUSION:P. falciparum and B. microti were highly sensitive to inactivation by PCT. Pathogen inactivation approaches could reduce the risk of transfusion-transmitted parasitic infections and avoid unnecessary donor exclusions.
- Published
- 2008
18. Protracted sterile protection with Plasmodium yoelii pre-erythrocytic genetically attenuated parasite malaria vaccines is independent of significant liver-stage persistence and is mediated by CD8+ T cells
- Author
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Ronald F. Dumpit, Stefan H. I. Kappe, Alice S. Tarun, Ruobing Wang, Akihide Takagi, Pu Liu, Mehdi Labaied, and Nelly Camargo
- Subjects
CD8-Positive T-Lymphocytes ,Vaccines, Attenuated ,Mice ,Immunity ,parasitic diseases ,Malaria Vaccines ,medicine ,Immunology and Allergy ,Animals ,Immunization Schedule ,Mice, Inbred BALB C ,Liver infection ,Attenuated vaccine ,biology ,Malaria vaccine ,Vaccination ,Plasmodium falciparum ,Plasmodium yoelii ,biology.organism_classification ,medicine.disease ,Virology ,Malaria ,Infectious Diseases ,Liver ,Sporozoites ,Immunology ,Injections, Intravenous ,Vacuoles ,Hepatocytes ,Female ,Gene Deletion - Abstract
Irradiation-attenuated sporozoite vaccinations confer sterile protection against malaria infection in animal models and humans. Persistent, nonreplicating parasite forms in the liver are presumably necessary for the maintenance of sterile immunity. A novel vaccine approach uses genetically attenuated parasites (GAPs) that undergo arrested development during liver infection. The fate of GAPs after immunization, their persistence in vaccinated animals, and the immune mechanisms that mediate protection are unknown. To examine the developmental defects of genetically attenuated liver stages in vivo, we created deletions of the UIS3 and UIS4 loci in the Plasmodium yoelii rodent malaria model (Pyuis3[-] and Pyuis4[-]). The low 50% infectious dose of P. yoelii in BALB/c mice provides the most sensitive infectivity model. We show that P. yoelii GAPs reach the liver, invade hepatocytes, and develop a parasitophorous vacuole but do not significantly persist 40 h after infection. A single dose of Pyuis4(-) sporozoites conferred complete protection, but full protection by Pyuis3(-) sporozoites required at least 2 immunizations. CD8(+) T cells were essential for protection, but CD4(+) T cells were not. Our results show that genetically distinct GAPs confer different degrees of protective efficacy and that live vaccine persistence in the liver is not necessary to sustain long-lasting protection. These findings have important implications for the development of a P. falciparum GAP malaria vaccine.
- Published
- 2006
19. Depletion of the Plasmodium berghei thrombospondin-related sporozoite protein reveals a role in host cell entry by sporozoites
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Nelly Camargo, Mehdi Labaied, and Stefan H. I. Kappe
- Subjects
Plasmodium berghei ,Molecular Sequence Data ,Protozoan Proteins ,Plasmodium ,Mice ,parasitic diseases ,medicine ,Parasite hosting ,Animals ,Humans ,Vector (molecular biology) ,Amino Acid Sequence ,Molecular Biology ,Gene ,Cells, Cultured ,Repetitive Sequences, Nucleic Acid ,Thrombospondin ,Liver infection ,biology ,fungi ,biology.organism_classification ,medicine.disease ,Virology ,Malaria ,Mice, Inbred C57BL ,Sporozoites ,Hepatocytes ,Parasitology ,Female ,Thrombospondins ,Gene Deletion - Abstract
The malaria parasite sporozoite stage develops in the mosquito vector and is transmitted to the mammalian host by bite. Sporozoites engage in multiple interactions with vector and host tissue on the journey from their oocyst origin to their final destination inside hepatocytes. Several malaria proteins have been identified that mediate sporozoite interactions with target tissues such as secreted and surface-associated ligands CSP and TRAP, which contain a thrombospondin type 1 repeat (TSR). Recently, we identified thrombospondin-related sporozoite protein (TRSP) in Plasmodium sporozoites, which exhibits a single TSR in its putative extracellular N-terminal region and is highly conserved among Plasmodium species. Here, we show using targeted gene disruption in the rodent malaria model Plasmodium berghei, that lack of TRSP has no effect on the asexual blood stage cycle, parasite transmission to the mosquito, sporozoite development and infection of mosquito salivary glands. However, analysis of TRSP knockout sporozoites in vitro and in vivo indicates that this protein has a significant role in hepatocyte entry and therefore liver infection. Thus, TRSP is an additional TSR-containing malaria parasite protein that is mainly involved in initial infection of the mammalian host.
- Published
- 2005
20. [Untitled]
- Author
-
Serge L. Y. Thomas, Shimon Gatt, Philippe Grellier, Stéphane Egée, M. Dellinger, Chunbo Wang, Marc Gèze, Arie Dagan, and Mehdi Labaied
- Subjects
0303 health sciences ,Ceramide ,biology ,030302 biochemistry & molecular biology ,Plasmodium falciparum ,biology.organism_classification ,Sphingolipid ,Plasmodium ,Cell biology ,carbohydrates (lipids) ,03 medical and health sciences ,chemistry.chemical_compound ,Infectious Diseases ,chemistry ,Biochemistry ,Cell culture ,parasitic diseases ,Sphingolipid metabolism ,lipids (amino acids, peptides, and proteins) ,Parasitology ,030304 developmental biology - Abstract
Background Sphingolipids are key molecules regulating many essential functions in eukaryotic cells and ceramide plays a central role in sphingolipid metabolism. A sphingolipid metabolism occurs in the intraerythrocytic stages of Plasmodium falciparum and is associated with essential biological processes. It constitutes an attractive and potential target for the development of new antimalarial drugs.
- Published
- 2004
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