Extracellular development, in vitro, of the erythrocytic cycle of plasmodium falciparum

1992 ◽  
Vol 8 (11) ◽  
pp. 384-387 ◽  
Author(s):  
W. Trager ◽  
J. Williams ◽  
G.S. Gill
Keyword(s):  
Plasmodium Falciparum ◽  
Erythrocytic Cycle ◽  
Development In Vitro

scholarly journals Overexpression of Plasmodium falciparum M1 Aminopeptidase Promotes an Increase in Intracellular Proteolysis and Modifies the Asexual Erythrocytic Cycle Development

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1452
Author(s):  
Carolina C. Hoff ◽  
Mauro F. Azevedo ◽  
Adriana B. Thurler ◽  
Sarah El Chamy Maluf ◽  
Pollyana M. S. Melo ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Antimalarial Activity ◽  
Fold Increase ◽  
Cysteine Proteases ◽  
Aminopeptidase Activity ◽  
Lower Sensitivity ◽  
Wild Type ◽  
Erythrocytic Cycle ◽  
Transgenic Parasite

Plasmodium falciparum, the most virulent of the human malaria parasite, is responsible for high mortality rates worldwide. We studied the M1 alanyl-aminopeptidase of this protozoan (PfA-M1), which is involved in the final stages of hemoglobin cleavage, an essential process for parasite survival. Aiming to help in the rational development of drugs against this target, we developed a new strain of P. falciparum overexpressing PfA-M1 without the signal peptide (overPfA-M1). The overPfA-M1 parasites showed a 2.5-fold increase in proteolytic activity toward the fluorogenic substrate alanyl-7-amido-4-methylcoumarin, in relation to the wild-type group. Inhibition studies showed that overPfA-M1 presented a lower sensitivity against the metalloaminopeptidase inhibitor bestatin and to other recombinant PfA-M1 inhibitors, in comparison with the wild-type strain, indicating that PfA-M1 is a target for the in vitro antimalarial activity of these compounds. Moreover, overPfA-M1 parasites present a decreased in vitro growth, showing a reduced number of merozoites per schizont, and also a decrease in the iRBC area occupied by the parasite in trophozoite and schizont forms when compared to the controls. Interestingly, the transgenic parasite displays an increase in the aminopeptidase activity toward Met-, Ala-, Leu- and Arg-7-amido-4-methylcoumarin. We also investigated the potential role of calmodulin and cysteine proteases in PfA-M1 activity. Taken together, our data show that the overexpression of PfA-M1 in the parasite cytosol can be a suitable tool for the screening of antimalarials in specific high-throughput assays and may be used for the identification of intracellular molecular partners that modulate their activity in P. falciparum.

scholarly journals The effect of purified aminoaldehydes produced by polyamine oxidation on the development in vitro of Plasmodium falciparum in normal and glucose-6-phosphate-dehydrogenase-deficient erythrocytes

1986 ◽  
Vol 236 (1) ◽  
pp. 97-101 ◽  
Author(s):  
D M L Morgan ◽  
U Bachrach ◽  
Y G Assaraf ◽  
E Harari ◽  
J Golenser
Keyword(s):  
Plasmodium Falciparum ◽  
Lethal Effect ◽  
Human Erythrocytes ◽  
Malarial Parasite ◽  
Parasite Plasmodium ◽  
Parasite Plasmodium Falciparum ◽  
Development In Vitro ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Polyamine Oxidation

Purified aminoaldehydes produced by polyamine oxidation were toxic to the malarial parasite, Plasmodium falciparum, cultured in human erythrocytes. There was a profound effect on young ring forms, and, during maturation, parasites became more sensitive to the aldehydes. Oxidation of the aldehydes abolished the lethal effect. The plasmodia within glucose-6-phosphate-dehydrogenase (G6PD)-deficient erythrocytes were more sensitive to mono- and di-aldehydes than were parasites in normal erythrocytes. G6PD-deficient erythrocytes were also more sensitive to pretreatment with the dialdehyde produced by the oxidation of spermine. Pretreatment prevented further invasion by the parasites.

Effects of the angiotensin II Ala-scan analogs in erythrocytic cycle of Plasmodium falciparum (in vitro) and Plasmodium gallinaceum (ex vivo)

2015 ◽  
Vol 153 ◽  
pp. 1-7 ◽  
Author(s):  
Adriana Farias Silva ◽  
Leandro de Souza Silva ◽  
Flávio Lopes Alves ◽  
Marcelo Der TorossianTorres ◽  
Ana Acacia de SáPinheiro ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Angiotensin Ii ◽  
Ex Vivo ◽  
Plasmodium Gallinaceum ◽  
Erythrocytic Cycle

A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum

2020 ◽  
Author(s):  
Kasem Kulkeaw ◽  
Nutpakal Ketprasit ◽  
Anchalee Tungtrongchitr ◽  
Duangdao Palasuwan
Keyword(s):  
Plasmodium Falciparum ◽  
Microscopic Examination ◽  
Inhibitory Effect ◽  
Nucleic Acid Binding ◽  
Flow Cytometric Assay ◽  
Malaria Therapy ◽  
Flow Cytometric ◽  
Development In Vitro ◽  
Vitro Development

Abstract Background Gold standard microscopic examination of Plasmodium falciparum intraerythrocytic stage remains an important process for staging and enumerating parasitized erythrocytes in culture; however, microscopy is laborious and its accuracy is dependent upon the skill of the examiner. Methods In this study, ViSafe Green (VSG), which is a nucleic acid-binding fluorescent dye, was used for assessing in vitro development of P. falciparum using flow cytometry. Results Fluorescence intensity of VSG was found to depend on the developmental stage of parasites. Specifically, multiple-nuclei-containing schizonts were observed in the VSGhigh population, and growing trophozoites and ring-shaped forms were observed in the VSGintermediate and VSGlow populations. The efficacy of VSG-based assay was found to be comparable to the microscopic examination method, and it demonstrated an ability to detect as low as 0.001% of the parasitaemia estimated by Giemsa staining. Moreover, when applying VSG for anti-malarial drug test, it was able to observe the growth inhibitory effect of dihydroartemisinin, the front-line drug for malaria therapy. Conclusions Taken together, the results of this study suggest the VSG-based flow cytometric assay to be a simple and reliable assay for assessing P. falciparum malaria development in vitro.

scholarly journals In Vitro and In Vivo Properties of Ellagic Acid in Malaria Treatment

2008 ◽  
Vol 53 (3) ◽  
pp. 1100-1106 ◽  
Author(s):  
Patrice Njomnang Soh ◽  
Benoît Witkowski ◽  
David Olagnier ◽  
Marie-Laure Nicolau ◽  
Maria-Concepcion Garcia-Alvarez ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Ellagic Acid ◽  
Antimalarial Activity ◽  
Inhibitory Effect ◽  
Oral Route ◽  
Mechanisms Of Action ◽  
The World ◽  
Erythrocytic Cycle

ABSTRACT Malaria is one of the most significant causes of infectious disease in the world. The search for new antimalarial chemotherapies has become increasingly urgent due to the parasites’ resistance to current drugs. Ellagic acid is a polyphenol found in various plant products. In this study, antimalarial properties of ellagic acid were explored. The results obtained have shown high activity in vitro against all Plasmodium falciparum strains whatever their levels of chloroquine and mefloquine resistance (50% inhibitory concentrations ranging from 105 to 330 nM). Ellagic acid was also active in vivo against Plamodium vinckei petteri in suppressive, curative, and prophylactic murine tests, without any toxicity (50% effective dose by the intraperitoneal route inferior to 1 mg/kg/day). The study of the point of action of its antimalarial activity in the erythrocytic cycle of Plasmodium falciparum demonstrated that it occurred at the mature trophozoite and young schizont stages. Moreover, ellagic acid has been shown to potentiate the activity of current antimalarial drugs such as chloroquine, mefloquine, artesunate, and atovaquone. This study also proved the antioxidant activity of ellagic acid and, in contrast, the inhibitory effect of the antioxidant compound N-acetyl-l-cysteine on its antimalarial efficacy. The possible mechanisms of action of ellagic acid on P. falciparum are discussed in light of the results. Ellagic acid has in vivo activity against plasmodia, but modification of the compound could lead to improved pharmacological properties, principally for the oral route.

scholarly journals Roles for Two Aminopeptidases in Vacuolar Hemoglobin Catabolism in Plasmodium falciparum

2007 ◽  
Vol 282 (49) ◽  
pp. 35978-35987 ◽  
Author(s):  
Seema Dalal ◽  
Michael Klemba
Keyword(s):  
Amino Acids ◽  
Plasmodium Falciparum ◽  
Current Model ◽  
Food Vacuole ◽  
Peptide Transport ◽  
Parasite Development ◽  
Aminopeptidase Activity ◽  
Erythrocytic Cycle ◽  
The Impact

During the erythrocytic stage of its life cycle, the human malaria parasite Plasmodium falciparum catabolizes large quantities of host-cell hemoglobin in an acidic organelle, the food vacuole. A current model for the catabolism of globin-derived oligopeptides invokes peptide transport out of the food vacuole followed by hydrolysis to amino acids by cytosolic aminopeptidases. To test this model, we have examined the roles of four parasite aminopeptidases during the erythrocytic cycle. Localization of tagged aminopeptidases, coupled with biochemical analysis of enriched food vacuoles, revealed the presence of amino acid-generating pathways in the food vacuole as well as the cytosol. Based on the localization data and in vitro assays, we propose a specific role for one of the plasmodial enzymes, aminopeptidase P, in the catabolism of proline-containing peptides in both the vacuole and the cytosol. We establish an apparent requirement for three of the four aminopeptidases (including the two food vacuole enzymes) for efficient parasite proliferation. To gain insight into the impact of aminopeptidase inhibition on parasite development, we examined the effect of the presence of amino acids in the culture medium of the parasite on the toxicity of the aminopeptidase inhibitor bestatin. The ability of bestatin to block parasite replication was only slightly affected when 19 of 20 amino acids were withdrawn from the medium, indicating that exogenous amino acids cannot compensate for the loss of aminopeptidase activity. Together, these results support the development of aminopeptidase inhibitors as novel chemotherapeutics directed against malaria.

scholarly journals Febrile temperatures can synchronize the growth of Plasmodium falciparum in vitro.

1989 ◽  
Vol 169 (1) ◽  
pp. 357-361 ◽  
Author(s):  
D Kwiatkowski
Keyword(s):  
Plasmodium Falciparum ◽  
Temperature Sensitivity ◽  
Natural Infection ◽  
Effect Of Temperature ◽  
Parasite Development ◽  
In Vitro Growth ◽  
Erythrocytic Cycle ◽  
Fever Response

To investigate the possibility that the host fever response in malaria may affect parasite development, we studied the effect of temperature on Plasmodium falciparum in erythrocytic culture in vitro. Growth was markedly suppressed at 40 degrees C compared with 37 degrees C, due to disruption of the second half of the 48-h erythrocytic cycle. However, young intraerythrocytic parasites, which are highly exposed to fever during natural infection, appeared to develop normally at 40 degrees C. Because of the differential temperature sensitivity within the erythrocytic cycle, asynchronous cultures could be synchronized by transient elevations of temperature. Pronounced synchronization was observed when cultures were exposed to periodic elevations of temperature that simulated the 48-h fever cycle of tertian malaria. These findings indicate that malaria fever might act to promote parasite synchronization in vivo.

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