RESUMEN
A fungal metabolite, diatretol, has shown to be a promising antimalarial agent. Diatretol displayed potent in vitro antiparasitic activity against the Plasmodium falciparum K1 strain, with an IC50 value of 378 ng ml-1, as well as in vivo efficacy in a Plasmodium berghei-infected mice model, with ca. 50% inhibition at 30 mg/kg (p.o.).
Asunto(s)
Antimaláricos/farmacología , Malaria/tratamiento farmacológico , Animales , Antimaláricos/química , Dicetopiperazinas/química , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Eritrocitos/parasitología , Humanos , Malaria/parasitología , Ratones Endogámicos ICR , Parasitemia/tratamiento farmacológico , Parasitemia/parasitología , Plasmodium berghei/parasitología , Plasmodium falciparum/efectos de los fármacosRESUMEN
New antimalarial therapeutics are needed to ensure that malaria cases continue to be driven down, as both emerging parasite resistance to frontline chemotherapies and mosquito resistance to current insecticides threaten control programmes. Plasmodium, the apicomplexan parasite responsible for malaria, causes disease pathology through repeated cycles of invasion and replication within host erythrocytes (the asexual cycle). Antimalarial drugs primarily target this cycle, seeking to reduce parasite burden within the host as fast as possible and to supress recrudescence for as long as possible. Intense phenotypic drug screening efforts have identified a number of promising new antimalarial molecules. Particularly important is the identification of compounds with new modes of action within the parasite to combat existing drug resistance and suitable for formulation of efficacious combination therapies. Here we detail the antimalarial properties of DDD01034957-a novel antimalarial molecule which is fast-acting and potent against drug resistant strains in vitro, shows activity in vivo, and possesses a resistance mechanism linked to the membrane transporter PfABCI3. These data support further medicinal chemistry lead-optimization of DDD01034957 as a novel antimalarial chemical class and provide new insights to further reduce in vivo metabolic clearance.
Asunto(s)
Antimaláricos/farmacología , Resistencia a Medicamentos/efectos de los fármacos , Malaria/tratamiento farmacológico , Plasmodium falciparum/efectos de los fármacos , Animales , Antimaláricos/química , Eritrocitos/parasitología , Interacciones Huésped-Parásitos/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Malaria/parasitología , Ratones , Estructura Molecular , Plasmodium/efectos de los fármacos , Plasmodium/parasitología , Plasmodium berghei/efectos de los fármacos , Plasmodium berghei/parasitología , Plasmodium falciparum/fisiología , Especificidad de la EspecieRESUMEN
The protozoan parasite Plasmodium, causative agent of malaria, invades hepatocytes by invaginating the host cell plasma membrane and forming a parasitophorous vacuole membrane (PVM). Surrounded by this PVM, the parasite undergoes extensive replication. Parasites inside a PVM provoke the Plasmodium-associated autophagy-related (PAAR) response. This is characterised by a long-lasting association of the autophagy marker protein LC3 with the PVM, which is not preceded by phosphatidylinositol 3-phosphate (PI3P)-labelling. Prior to productive invasion, sporozoites transmigrate several cells and here we describe that a proportion of traversing sporozoites become trapped in a transient traversal vacuole, provoking a host cell response that clearly differs from the PAAR response. These trapped sporozoites provoke PI3P-labelling of the surrounding vacuolar membrane immediately after cell entry, followed by transient LC3-labelling and elimination of the parasite by lysosomal acidification. Our data suggest that this PI3P response is not only restricted to sporozoites trapped during transmigration but also affects invaded parasites residing in a compromised vacuole. Thus, host cells can employ a pathway distinct from the previously described PAAR response to efficiently recognise and eliminate Plasmodium parasites.
Asunto(s)
Autofagia , Hepatocitos/parasitología , Fosfatos de Fosfatidilinositol/metabolismo , Plasmodium berghei/metabolismo , Plasmodium berghei/parasitología , Esporozoítos/metabolismo , Vacuolas/parasitología , Animales , Línea Celular , Femenino , Células HeLa , Interacciones Huésped-Parásitos , Humanos , Malaria/parasitología , Ratones , Proteínas Asociadas a Microtúbulos/metabolismo , Organismos Modificados GenéticamenteRESUMEN
BACKGROUND: Malaria is a parasitic disease that compromises the human host. Currently, control of the Plasmodium falciparum burden is centered on artemisinin-based combination therapies. However, decreased sensitivity to artemisinin and derivatives has been reported, therefore it is important to identify new therapeutic strategies. METHOD: We used human erythrocytes infected with P. falciparum and experimental cerebral malaria (ECM) animal model to assess the potential antimalarial effect of eugenol, a component of clove bud essential oil. RESULTS: Plasmodium falciparum cultures treated with increasing concentrations of eugenol reduced parasitemia in a dose-dependent manner, with IC50 of 532.42 ± 29.55 µM. This effect seems to be irreversible and maintained even in the presence of high parasitemia. The prominent effect of eugenol was detected in the evolution from schizont to ring forms, inducing important morphological changes, indicating a disruption in the development of the erythrocytic cycle. Aberrant structural modification was observed by electron microscopy, showing the separation of the two nuclear membrane leaflets as well as other subcellular membranes, such as from the digestive vacuole. Importantly, in vivo studies using ECM revealed a reduction in blood parasitemia and cerebral edema when mice were treated for 6 consecutive days upon infection. CONCLUSIONS: These data suggest a potential effect of eugenol against Plasmodium sp. with an impact on cerebral malaria. GENERAL SIGNIFICANCE: Our results provide a rational basis for the use of eugenol in therapeutic strategies to the treatment of malaria.
Asunto(s)
Antimaláricos/farmacología , Edema Encefálico/tratamiento farmacológico , Eugenol/farmacología , Estadios del Ciclo de Vida/efectos de los fármacos , Malaria Cerebral/tratamiento farmacológico , Malaria Falciparum/tratamiento farmacológico , Plasmodium falciparum/efectos de los fármacos , Animales , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/metabolismo , Barrera Hematoencefálica/parasitología , Edema Encefálico/parasitología , Modelos Animales de Enfermedad , Eritrocitos/efectos de los fármacos , Eritrocitos/parasitología , Humanos , Concentración 50 Inhibidora , Estadios del Ciclo de Vida/fisiología , Malaria Cerebral/parasitología , Malaria Falciparum/parasitología , Masculino , Ratones , Ratones Endogámicos C57BL , Plasmodium berghei/efectos de los fármacos , Plasmodium berghei/crecimiento & desarrollo , Plasmodium berghei/parasitología , Plasmodium falciparum/crecimiento & desarrollo , Plasmodium falciparum/patogenicidadRESUMEN
Malaria, the disease caused by Plasmodium spp. infection, remains a major global cause of morbidity and mortality. Host protection from malaria relies on immune-driven resistance mechanisms that kill Plasmodium However, these mechanisms are not sufficient per se to avoid the development of severe forms of disease. This is accomplished instead via the establishment of disease tolerance to malaria, a defense strategy that does not target Plasmodium directly. Here we demonstrate that the establishment of disease tolerance to malaria relies on a tissue damage-control mechanism that operates specifically in renal proximal tubule epithelial cells (RPTEC). This protective response relies on the induction of heme oxygenase-1 (HMOX1; HO-1) and ferritin H chain (FTH) via a mechanism that involves the transcription-factor nuclear-factor E2-related factor-2 (NRF2). As it accumulates in plasma and urine during the blood stage of Plasmodium infection, labile heme is detoxified in RPTEC by HO-1 and FTH, preventing the development of acute kidney injury, a clinical hallmark of severe malaria.
Asunto(s)
Hemo/metabolismo , Riñón/metabolismo , Malaria/fisiopatología , Animales , Apoferritinas/metabolismo , Línea Celular , Progresión de la Enfermedad , Células Epiteliales/metabolismo , Ferritinas/metabolismo , Ferritinas/fisiología , Hemo-Oxigenasa 1/metabolismo , Hemo-Oxigenasa 1/fisiología , Humanos , Tolerancia Inmunológica/fisiología , Ratones , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2/metabolismo , Factor 2 Relacionado con NF-E2/fisiología , Oxidorreductasas , Plasmodium berghei/metabolismo , Plasmodium berghei/parasitología , Regulación hacia ArribaRESUMEN
Within the liver a single Plasmodium parasite transforms into thousands of blood-infective forms to cause malaria. Here, we use RNA-sequencing to identify host genes that are upregulated upon Plasmodium berghei infection of hepatocytes with the hypothesis that host pathways are hijacked to benefit parasite development. We found that expression of aquaporin-3 (AQP3), a water and glycerol channel, is significantly induced in Plasmodium-infected hepatocytes compared to uninfected cells. This aquaglyceroporin localizes to the parasitophorous vacuole membrane, the compartmental interface between the host and pathogen, with a temporal pattern that correlates with the parasite's expansion in the liver. Depletion or elimination of host AQP3 expression significantly reduces P. berghei parasite burden during the liver stage and chemical disruption by a known AQP3 inhibitor, auphen, reduces P. falciparum asexual blood stage and P. berghei liver stage parasite load. Further use of this inhibitor as a chemical probe suggests that AQP3-mediated nutrient transport is an important function for parasite development. This study reveals a previously unknown potential route for host-dependent nutrient acquisition by Plasmodium which was discovered by mapping the transcriptional changes that occur in hepatocytes throughout P. berghei infection. The dataset reported may be leveraged to identify additional host factors that are essential for Plasmodium liver stage infection and highlights Plasmodium's dependence on host factors within hepatocytes.
Asunto(s)
Acuaporina 3/metabolismo , Plasmodium berghei/metabolismo , Animales , Acuaporina 3/fisiología , Células Hep G2 , Hepatocitos/metabolismo , Hepatocitos/parasitología , Humanos , Hígado/metabolismo , Hígado/parasitología , Hepatopatías , Malaria/parasitología , Ratones , Parásitos/metabolismo , Plasmodium berghei/genética , Plasmodium berghei/parasitología , Proteínas Protozoarias/metabolismo , Análisis de Secuencia de ARN/métodos , Esporozoítos/metabolismo , Vacuolas/metabolismoRESUMEN
Tissue macrophages exhibit diverse functions, ranging from the maintenance of tissue homeostasis, including clearance of senescent erythrocytes and cell debris, to modulation of inflammation and immunity. Their contribution to the control of blood-stage malaria remains unclear. Here, we show that in the absence of tissue-resident CD169(+) macrophages, Plasmodium berghei ANKA (PbA) infection results in significantly increased parasite sequestration, leading to vascular occlusion and leakage and augmented tissue deposition of the malarial pigment hemozoin. This leads to widespread tissue damage culminating in multiple organ inflammation. Thus, the capacity of CD169(+) macrophages to contain the parasite burden and its sequestration into different tissues and to limit infection-induced inflammation is crucial to mitigating Plasmodium infection and pathogenesis.
Asunto(s)
Macrófagos/parasitología , Malaria/inmunología , Plasmodium berghei/parasitología , Lectina 1 Similar a Ig de Unión al Ácido Siálico/genética , Animales , Eritrocitos/parasitología , Hemoproteínas/metabolismo , Macrófagos/patología , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones TransgénicosRESUMEN
Malarial infection is initiated when the sporozoite form of the Plasmodium parasite is inoculated into the skin by a mosquito. Sporozoites invade hepatocytes in the liver and develop into the erythrocyte-infecting form of the parasite, the cause of clinical blood infection. Protection against parasite development in the liver can be induced by injection of live attenuated parasites that do not develop in the liver and thus do not cause blood infection. Radiation-attenuated sporozoites (RAS) and genetically attenuated parasites are now considered as lead candidates for vaccination of humans against malaria. Although the skin appears as the preferable administration route, most studies in rodents, which have served as model systems, have been performed after i.v. injection of attenuated sporozoites. In this study, we analyzed the early response to Plasmodium berghei RAS or wild-type sporozoites (WTS) injected intradermally into C57BL/6 mice. We show that RAS have a similar in vivo distribution to WTS and that both induce a similar inflammatory response consisting of a biphasic recruitment of polymorphonuclear neutrophils and inflammatory monocytes in the skin injection site and proximal draining lymph node (dLN). Both WTS and RAS associate with neutrophils and resident myeloid cells in the skin and the dLN, transform inside CD11b(+) cells, and induce a Th1 cytokine profile in the dLN. WTS and RAS are also similarly capable of priming parasite-specific CD8(+) T cells. These studies delineate the early and local response to sporozoite injection into the skin, and suggest that WTS and RAS prime the host immune system in a similar fashion.
Asunto(s)
Malaria/inmunología , Plasmodium berghei/inmunología , Piel/inmunología , Piel/parasitología , Esporozoítos/inmunología , Esporozoítos/trasplante , Animales , Anopheles/inmunología , Anopheles/parasitología , Presentación de Antígeno/inmunología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/parasitología , Línea Celular , Oído , Femenino , Células Hep G2 , Interacciones Huésped-Parásitos/inmunología , Humanos , Inflamación/inmunología , Inflamación/parasitología , Inflamación/patología , Inyecciones Intradérmicas , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/parasitología , Malaria/parasitología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Plasmodium berghei/parasitología , Piel/citología , Esporozoítos/efectos de la radiaciónRESUMEN
BACKGROUND: Plasmodium berghei ANKA infection in C57Bl/6 mice induces cerebral malaria (CM), which reproduces, to a large extent, the pathological features of human CM. However, experimental CM incidence is variable (50-100%) and the period of incidence may present a range as wide as 6-12 days post-infection. The poor predictability of which and when infected mice will develop CM can make it difficult to determine the causal relationship of early pathological changes and outcome. With the purpose of contributing to solving these problems, algorithms for CM prediction were built. METHODS: Seventy-eight P. berghei-infected mice were daily evaluated using the primary SHIRPA protocol. Mice were classified as CM+ or CM- according to development of neurological signs on days 6-12 post-infection. Logistic regression was used to build predictive models for CM based on the results of SHIRPA tests and parasitaemia. RESULTS: The overall CM incidence was 54% occurring on days 6-10. Some algorithms had a very good performance in predicting CM, with the area under the receiver operator characteristic ((au)ROC) curve > or = 80% and positive predictive values (PV+) > or = 95, and correctly predicted time of death due to CM between 24 and 72 hours before development of the neurological syndrome ((au)ROC = 77-93%; PV+ = 100% using high cut off values). Inclusion of parasitaemia data slightly improved algorithm performance. CONCLUSION: These algorithms work with data from a simple, inexpensive, reproducible and fast protocol. Most importantly, they can predict CM development very early, estimate time of death, and might be a valuable tool for research using CM murine models.
Asunto(s)
Algoritmos , Conducta Animal/fisiología , Malaria Cerebral/patología , Examen Neurológico , Plasmodium berghei/patogenicidad , Animales , Modelos Animales de Enfermedad , Femenino , Predicción , Modelos Logísticos , Ratones , Ratones Endogámicos C57BL , Enfermedades del Sistema Nervioso/genética , Pruebas Neuropsicológicas , Plasmodium berghei/parasitologíaRESUMEN
Successful development of Plasmodium sexual stages is essential for parasite survival, but the genes involved are poorly understood. We 'knocked out' the male development gene-1 (mdv-1) locus in Plasmodium berghei and found it to be important in female gametocyte activation. Indirect immunofluorescence assays show MDV-1 has a punctate cytoplasmic distribution in gametocytes. After activation of both females and males, MDV-1 is more peripherally located but in males exclusively it becomes concentrated in a few large foci. In vitro ookinete conversion assays that test the ability of activated female gametocytes to develop into retort stage ookinetes, suggests a complicit role for MDV-1, with the knock-out parasite producing 86% reduction in ookinetes. The retort stage ookinete develops from the zygote by increasing growth of an apical protrusion and MDV-1 locates at the 'leading' extracellular apical pole of this protrusion. In the fully developed ookinete MDV-1 is localised to the posterior pole. In vivo, the knock-out parasites demonstrate a phenotype in which there is a 90% reduction of parasite transmission to oocysts in mosquitoes.
Asunto(s)
Membrana Celular/metabolismo , Gametogénesis/fisiología , Plasmodium berghei/crecimiento & desarrollo , Proteínas Protozoarias/metabolismo , Desarrollo Sexual , Cigoto/crecimiento & desarrollo , Secuencia de Aminoácidos , Animales , Animales Modificados Genéticamente , Anopheles/parasitología , Western Blotting , Proteínas de Ciclo Celular/metabolismo , Membrana Celular/parasitología , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Eliminación de Gen , Genes del Desarrollo/fisiología , Masculino , Ratones , Datos de Secuencia Molecular , Oocistos/crecimiento & desarrollo , Oocistos/fisiología , Fenotipo , Plasmodium berghei/genética , Plasmodium berghei/parasitología , Proteoma/fisiología , Proteínas Protozoarias/genética , Conejos , Razón de Masculinidad , Cigoto/metabolismoRESUMEN
Plasmodium parasites, the causal agents of malaria, dramatically modify the infected erythrocyte by exporting parasite proteins into one or multiple erythrocyte compartments, the cytoplasm and the plasma membrane or beyond. Despite advances in defining signals and specific cellular compartments implicated in protein trafficking in Plasmodium-infected erythrocytes, the contribution of lipid-mediated sorting to this cellular process has been poorly investigated. In this study, we examined the proteome of cholesterol-rich membrane microdomains or lipid rafts, purified from erythrocytes infected by the rodent parasite Plasmodium berghei. Besides structural proteins associated with invasive forms, we detected chaperones, proteins implicated in vesicular trafficking, membrane fusion events and signalling. Interestingly, the raft proteome of mixed P. berghei blood stages included proteins encoded by members of a large family (bir) of putative variant antigens potentially implicated in host immune system interactions and targeted to the surface of the host erythrocytes. The generation of transgenic parasites expressing BIR/GFP fusions confirmed the dynamic association of members of this protein family with membrane microdomains. Our results indicated that lipid rafts in Plasmodium-infected erythrocytes might constitute a route to sort and fold parasite proteins directed to various host cell compartments including the cell surface.
Asunto(s)
Antígenos/inmunología , Microdominios de Membrana/química , Plasmodium/fisiología , Proteínas Protozoarias/metabolismo , Transducción de Señal , Vesículas Transportadoras/metabolismo , Animales , Antígenos/genética , Eritrocitos/metabolismo , Eritrocitos/parasitología , Interacciones Huésped-Parásitos , Humanos , Estadios del Ciclo de Vida , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos , Plasmodium/química , Plasmodium/genética , Plasmodium/metabolismo , Plasmodium berghei/genética , Plasmodium berghei/crecimiento & desarrollo , Plasmodium berghei/parasitología , Transporte de Proteínas , Proteómica/métodos , Proteínas Protozoarias/análisis , Tripsina/farmacologíaRESUMEN
Limitações atuais do arsenal terapêutico na malária humana exigem a busca de novos medicamentos. Além da grande importância econômica e social da malária e da resistência do P. falciparum a cloroquina e outros derivados quinolínicos, as associações medicamentosas compostas por artemisinina e seus derivados apresentam custos de produção elevados, dificultando seu emprego em regiões em desenvolvimento, especialmente no continente africano. Nesse trabalho testamos a atividade antimalárica de: (i) associações dos antimaláricos artesunato (AS) e mefloquina (MQ) com ciprofloxacina, uma fluoroquinolona sintética empregada contra infecções bacterianas; (ii) moléculas obtidas por síntese química, como anti-retrovrais, endoperóxidos, e uma nova molécula híbrida denominada MEFAS e (iii) extratos e compostos purificados de plantas medicinais como falsas quinas e Holostylis reniformis, utilizadas popularmente no tratamento de quadros febris e dispepsias. Os testes esquizonticidas foram feitos em cultivos contínuos de P. falciparum avaliando-se o crescimento das fases intraeritrocitárias através de duas metodologias (teste tradicional ou o ensaio de incorporação de hipoxantina tritiada); e utilizando camundongos inoculados com P. berghei. Estabelecemos ainda um protocolo com sondas fluorescentes que permitiu trabalhar com trofozoítos de P. falciparum viáveis em eritrócitos infectados e acompanhar, em tempo real, ações de potenciais antimaláricos sobre a homeostasia iônica dos parasitos, identificando possíveis alvos intracelulares dos mesmos.
Asunto(s)
Ciprofloxacina/farmacología , Malaria Falciparum/tratamiento farmacológico , Plantas Medicinales , Plasmodium berghei/parasitología , Plasmodium falciparum/parasitologíaRESUMEN
Methods facilitating research in malaria are of pivotal relevance. Flow cytometry offers the possibility of rapid enumeration of parasitemia. It relies on staining the parasite DNA to distinguish between infected and non-infected red blood cell (RBC) populations. Unfortunately, in rodents abundant reticulocyte RNA interferes with the application of the method. This results in time-consuming sample preparation protocols that offer no clear advantage over microscopic counting. We re-evaluated the use of the DNA/RNA discriminating vital fluorochrome acridine orange (AO) for rapid flow cytometric enumeration of parasitemia in rodents. Whole blood from rodents infected with Plasmodium berghei and Plasmodium yoelii was stained with AO and analyzed by flow cytometer. A newly developed two-channel (FL1/FL3) detection method was compared with conventional one-channel (FL1) detection and microscopic counting. The new AO two-channel detection method clearly discriminated between infected and non-infected RBC populations. It showed to be linear above parasitemias of 0.3%. Sample processing time amounted to approximately 5 min. It is shown that AO can be used for rapid, precise, and accurate enumeration of parasitemia in rodents. Due to its ease of handling the method might find widespread application in malaria research.
Asunto(s)
Naranja de Acridina , Eritrocitos/parasitología , Citometría de Flujo/métodos , Colorantes Fluorescentes , Malaria/parasitología , Plasmodium berghei/aislamiento & purificación , Plasmodium yoelii/aislamiento & purificación , Animales , Cricetinae , Malaria/diagnóstico , Mesocricetus , Ratones , Ratones Endogámicos ICR , Parasitemia/parasitología , Plasmodium berghei/parasitología , Plasmodium yoelii/parasitología , Coloración y EtiquetadoRESUMEN
Invasion of hepatocytes by Plasmodium sporozoites is a prerequisite for establishment of a natural malaria infection. The molecular mechanisms underlying sporozoite invasion are largely unknown. We have previously reported that infection by Plasmodium falciparum and Plasmodium yoelii sporozoites depends on CD81 and cholesterol-dependent tetraspanin-enriched microdomains (TEMs) on the hepatocyte surface. Here we have analyzed the role of CD81 and TEMs during infection by sporozoites from the rodent parasite Plasmodium berghei. We found that depending on the host cell type, P. berghei sporozoites can use several distinct pathways for invasion. Infection of human HepG2, HuH7 and HeLa cells by P. berghei does not depend on CD81 or host membrane cholesterol, whereas both CD81 and cholesterol are required for infection of mouse hepatoma Hepa1-6 cells. In primary mouse hepatocytes, both CD81-dependent and -independent mechanisms participate in P. berghei infection and the relative contribution of the different pathways varies, depending on mouse genetic background. The existence of distinct invasion pathways may explain why P. berghei sporozoites are capable of infecting a wide range of host cell types in vitro. It could also provide a means for human parasites to escape immune responses and face polymorphisms of host receptors. This may have implications for the development of an anti-malarial vaccine targeting sporozoites.
Asunto(s)
Antígenos CD/fisiología , Hepatocitos/parasitología , Plasmodium berghei/patogenicidad , Esporozoítos/fisiología , Animales , Línea Celular , Colesterol/fisiología , Hepatocitos/metabolismo , Interacciones Huésped-Parásitos , Humanos , Proteínas de la Membrana/fisiología , Ratones , Plasmodium berghei/parasitología , Ratas , Tetraspanina 28 , TetraspaninasRESUMEN
Extracts from thirteen species of plants were evaluated by "in vivo" antimalarial test against plasmodium berghei effects. Significant activities were observed in the ethyl acetate and aqueous extracts, elaborated of Cedrela tonduzii leaves, Trichilia havanensis and Trichilia americana barks, Neurolaena lobata and Gliricidia sepium leaves and Duranta repens fruits. Compounds identified include flavanoids, coumarins, mellilotic acid and iridoids which some kind of biodynamic activity has previously been reported. The flavone quercetin 1 purified from C. tonduzii gave strong antimalarial activity, however, its respective glucosides (quercetin 3-glucoside 2 y robinine 7) showed little significant activity.
Asunto(s)
Antimaláricos/uso terapéutico , Cumarinas/uso terapéutico , Flavonoides/uso terapéutico , Glucósidos/uso terapéutico , Malaria/tratamiento farmacológico , Extractos Vegetales/química , Extractos Vegetales/farmacología , Plasmodium berghei/efectos de los fármacos , Animales , Antimaláricos/farmacología , Cumarinas/farmacología , Flavonoides/farmacología , Glucósidos/farmacología , Malaria/parasitología , Ratones , Plasmodium berghei/parasitologíaRESUMEN
Extracts from thirteen species of plants were evaluated by(r)in vivo(r)antimalarial test against plasmodium berghei effects. Significant activities were observed in the ethyl acetate and aqueous extracts, elaborated of Cedrela tonduzii leaves, Trichilia havanensis and Trichilia americana barks, Neurolaena lobata and liricidia sepium leaves and Duranta repens fruits. Compounds identified include flavanoids, coumarins, mellilotic acid and iridoids which some kind of biodynamic activity has previously been reported. The flavone uercetin 1 purified from C. tonduzii gave strong antimalarial activity, however, its respective glucosides (quercetin 3-glucoside 2 y robinine 7) showed little significant activity
Asunto(s)
Animales , Antimaláricos/uso terapéutico , Cumarinas/uso terapéutico , Flavonoides/uso terapéutico , Glucósidos/uso terapéutico , Malaria/tratamiento farmacológico , Extractos Vegetales/química , Extractos Vegetales/farmacología , Plasmodium berghei/efectos de los fármacos , Antimaláricos/farmacología , Cumarinas/farmacología , Flavonoides/farmacología , Malaria/parasitología , Extractos Vegetales , Plasmodium berghei/parasitologíaRESUMEN
Evidencias clinicas e experimentais sugerem que hospedeiros com deficiencia de ferro sao menos susceptiveis a malaria grave e que a suplementacao de ferro poderia agravar a infeccao na tentativa de justificar estes fatos, dois experimentos foram realizados. Os resultados obtidos sugerem que os animais com deficiencia de ferro apresentam clareamento das parasitemias mais tardio e que o desenvolvimento de P. berghei em ratos nao e suprimido pela deficiencia de ferro. A suplementacao de ferro antes e durante a infeccao nao aumentou as parasitemias. Alem disso, a replecao de ferro durante a infeccao produziu incrementos em parametros hematologicos em animais com anemia ferropriva
Asunto(s)
Animales , Masculino , Dieta , Hierro/administración & dosificación , Hierro/farmacología , Malaria/fisiopatología , Plasmodium berghei/efectos de los fármacos , Plasmodium berghei/parasitología , Ratas Wistar , Anemia Ferropénica , Bioensayo , Minerales en la Dieta , Fenómenos Fisiológicos Nutricionales del LactanteRESUMEN
Mössbauer studies of rat erythrocytes infected by Plasmodium berghei malaria parasites, using 57Fe-enriched rat red blood cells, were carried out in order to determine the physical parameters which characterize the malarial pigment iron and to test the effect of the widely used antimalaria drug, chloroquine, on these parameters. The iron in the malarial pigment which is derived from hemoglobin digestion by the intracellular parasite was found to be trivalent, high spin, with Mössbauer parameters which are significantly different from those of any known iron porphyrin containing compound. No difference was found between the parameters obtained in erythrocytes infected by drug-sensitive and drug-resistant strains of P. berghei, both before and after the treatment with chloroquine. The iron compound consists of microaggregates, about 30 A in diameter. These are somewhat larger in chloroquine-resistant strains and tend to increase in size in chloroquine-sensitive strains upon treatment with the drug. Mössbauer spectra of erythrocytes infected by a chloroquine-resistant strain revealed pigment iron in relative amounts invariable of those found in chloroquine-sensitive strains, demonstrating that drug-resistant parasites indeed digest hemoglobin.
Asunto(s)
Cloroquina/farmacología , Eritrocitos/metabolismo , Malaria/sangre , Pigmentos Biológicos/sangre , Animales , Eritrocitos/efectos de los fármacos , Hierro/sangre , Radioisótopos de Hierro , Cinética , Plasmodium berghei/parasitología , Ratas , Ratas Endogámicas , Análisis Espectral/métodos , TemperaturaRESUMEN
When inoculated with sporozoites of Plasmodium berghei, a line of hepatoma cells (HepG2-A16) derived from human liver supports the complete asexual developmental cycle of the exoerythrocytic stage. Parasites were shown to resemble parasites in vivo in hepatocytes. Subinoculation of merozoites into mice induced a red blood cell infection.
Asunto(s)
Neoplasias Hepáticas Experimentales/parasitología , Malaria/parasitología , Plasmodium berghei/parasitología , Animales , Eritrocitos/parasitología , Técnicas In Vitro , Ratones , Ratones EndogámicosRESUMEN
The attachment and entry of Plasmodium berghei sporozoites to cultured human lung fibroblast (WI38) or hepatoma (HepG2-A16) cells in vitro has been visualized using an immunoperoxidase technique coupled with light microscopy. Attachment and entry was substantially more frequent with HepG2-A16 cells, and appeared to be mediated by the Pb44 sporozoite surface protective antigen. When sporozoites were incubated with intact monoclonal antibodies to Pb44 or their monovalent Fab fragments, attachment was inhibited, suggesting that this technique may be an in vitro assay of protective immunity. Sporozoites appeared to enter cells actively, rather than by cell phagocytosis. Once within a membrane-bound vacuole, the sporozoite transformed into a trophozoite. It was suggested that Pb44 recognized a specific cell receptor, and that this technique may permit receptor characterization.