RESUMO
Chemical conversion of the extract of natural resources is a very attractive way to expand the chemical space to discover bioactive compounds. In order to search for new medicines to treat parasitic diseases that cause high morbidity and mortality in affected countries in the world, the ethyl acetate extract from the rhizome of Alpinia galanga (L.) has been chemically converted by epoxidation using dioxirane generated in situ. The biological activity of chemically converted extract (CCE) of A. galanga (L.) significantly increased the activity against Leishmania major up to 82.6 ± 6.2 % at 25 µg/mL (whereas 2.7 ± 0.8% for the original extract). By bioassay-guided fractionation, new phenylpropanoids (1-6) and four known compounds, hydroquinone (7), 4-hydroxy(4-hydroxyphenyl)methoxy)benzaldehyde (8), isocoumarin cis 4-hydroxymelein (9), and (2S,3S,6R,7R,9S,10S)-humulene triepoxide (10) were isolated from CCE. The structures of isolated compounds were determined by spectroscopic analyses of 1D and 2D NMR, IR, and MS spectra. The most active compound was hydroquinone (7) with IC50 = 0.37 ± 1.37 µg/mL as a substantial active principle of CCE. In addition, the new phenylpropanoid 2 (IC50 = 27.8 ± 0.34 µg/mL) also showed significant activity against L. major compared to the positive control miltefosine (IC50 = 7.47 ± 0.3 µg/mL). The activities of the isolated compounds were also evaluated against Plasmodium falciparum, Trypanosoma brucei gambisense and Trypanosoma brucei rhodeisense. Interestingly, compound 2 was selectively active against trypanosomes with potent activity. To the best of our knowledge, this is the first report on the bioactive "unnatural" natural products from the crude extract of A. galanga (L.) by chemical conversion and on its activities against causal pathogens of leishmaniasis, trypanosomiasis, and malaria.
Assuntos
Alpinia/química , Antimaláricos , Extratos Vegetais/química , Plasmodium falciparum/crescimento & desenvolvimento , Propanóis , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/crescimento & desenvolvimento , Antimaláricos/química , Antimaláricos/isolamento & purificação , Antimaláricos/farmacologia , Propanóis/química , Propanóis/isolamento & purificação , Propanóis/farmacologia , Tripanossomicidas/química , Tripanossomicidas/isolamento & purificação , Tripanossomicidas/farmacologiaRESUMO
The protozoan diseases Human African Trypanosomiasis (HAT), Chagas disease (CD), and leishmaniases span worldwide and therefore their impact is a universal concern. The present regimen against kinetoplastid protozoan infections is poor and insufficient. Target-based design expands the horizon of drug design and development and offers novel chemical entities and potential drug candidates to the therapeutic arsenal against the aforementioned neglected diseases. In this review, we report the most promising targets of the main kinetoplastid parasites, as well as their corresponding inhibitors. This overview is part of the Special Issue, entitled "Advances of Medicinal Chemistry against Kinetoplastid Protozoa (Trypanosoma brucei, Trypanosoma cruzi and Leishmania spp.) Infections: Drug Design, Synthesis and Pharmacology".
Assuntos
Antiprotozoários/farmacologia , Doença de Chagas/tratamento farmacológico , Desenho de Fármacos , Leishmaniose/tratamento farmacológico , Terapia de Alvo Molecular/métodos , Tripanossomíase Africana/tratamento farmacológico , Animais , Antiprotozoários/síntese química , Antiprotozoários/classificação , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Descoberta de Drogas , Humanos , Insetos Vetores/efeitos dos fármacos , Insetos Vetores/parasitologia , Leishmania/efeitos dos fármacos , Leishmania/genética , Leishmania/crescimento & desenvolvimento , Leishmania/metabolismo , Leishmaniose/parasitologia , Leishmaniose/transmissão , Estágios do Ciclo de Vida/efeitos dos fármacos , Estágios do Ciclo de Vida/genética , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Estrutura Molecular , Proteínas de Protozoários/antagonistas & inibidores , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Relação Estrutura-Atividade , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei gambiense/genética , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei gambiense/metabolismo , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/genética , Trypanosoma cruzi/crescimento & desenvolvimento , Trypanosoma cruzi/metabolismo , Tripanossomíase Africana/parasitologia , Tripanossomíase Africana/transmissãoRESUMO
Human apolipoprotein L1 (APOL1) kills African trypanosomes except Trypanosoma rhodesiense and Trypanosoma gambiense, the parasites causing sleeping sickness. APOL1 uptake into trypanosomes is favoured by its association with the haptoglobin-related protein-haemoglobin complex, which binds to the parasite surface receptor for haptoglobin-haemoglobin. As haptoglobin-haemoglobin can saturate the receptor, APOL1 uptake is increased in haptoglobin-poor (hypohaptoglobinaemic) serum (HyHS). While T. rhodesiense resists APOL1 by RNA polymerase I (pol-I)-mediated expression of the serum resistance-associated (SRA) protein, T. gambiense resists by pol-II-mediated expression of the T. gambiense-specific glycoprotein (TgsGP). Moreover, in T. gambiense resistance to HyHS is linked to haptoglobin-haemoglobin receptor inactivation by mutation. We report that unlike T. gambiense, T. rhodesiense possesses a functional haptoglobin-haemoglobin receptor, and that like T. gambiense experimentally provided with active receptor, this parasite is killed in HyHS because of receptor-mediated APOL1 uptake. However, T. rhodesiense could adapt to low haptoglobin by increasing transcription of SRA. When assayed in Trypanosoma brucei, resistance to HyHS occurred with pol-I-, but not with pol-II-mediated SRA expression. Similarly, T. gambiense provided with active receptor acquired resistance to HyHS only when TgsGP was moved to a pol-I locus. Thus, transcription by pol-I favours adaptive gene regulation, explaining the presence of SRA in a pol-I locus.
Assuntos
Apolipoproteínas/toxicidade , Regulação da Expressão Gênica , Lipoproteínas HDL/toxicidade , RNA Polimerase I/metabolismo , Transcrição Gênica , Trypanosoma brucei rhodesiense/efeitos dos fármacos , Trypanosoma brucei rhodesiense/fisiologia , Adaptação Fisiológica , Apolipoproteína L1 , Haptoglobinas/análise , Humanos , Glicoproteínas de Membrana/biossíntese , Receptores de Superfície Celular/metabolismo , Soro/química , Trypanosoma brucei brucei/efeitos dos fármacos , Trypanosoma brucei brucei/genética , Trypanosoma brucei brucei/crescimento & desenvolvimento , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei gambiense/genética , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/genética , Trypanosoma brucei rhodesiense/crescimento & desenvolvimentoRESUMO
African sleeping sickness is a neglected tropical disease transmitted by tsetse flies. New and better drugs are still needed especially for its second stage, which is fatal if untreated. 28DAP010, a dipyridylbenzene analogue of DB829, is the second simple diamidine found to cure mice with central nervous system infections by a parenteral route of administration. 28DAP010 showed efficacy similar to that of DB829 in dose-response studies in mouse models of first- and second-stage African sleeping sickness. The in vitro time to kill, determined by microcalorimetry, and the parasite clearance time in mice were shorter for 28DAP010 than for DB829. No cross-resistance was observed between 28DAP010 and pentamidine on the tested Trypanosoma brucei gambiense isolates from melarsoprol-refractory patients. 28DAP010 is the second promising preclinical candidate among the diamidines for the treatment of second-stage African sleeping sickness.
Assuntos
Amidinas/farmacologia , Piridinas/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei rhodesiense/efeitos dos fármacos , Tripanossomíase Africana/tratamento farmacológico , Amidinas/síntese química , Amidinas/farmacocinética , Animais , Relação Dose-Resposta a Droga , Feminino , Humanos , Melarsoprol/farmacocinética , Melarsoprol/farmacologia , Camundongos , Pentamidina/farmacocinética , Pentamidina/farmacologia , Piridinas/síntese química , Piridinas/farmacocinética , Relação Estrutura-Atividade , Tripanossomicidas/síntese química , Tripanossomicidas/farmacocinética , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei gambiense/patogenicidade , Trypanosoma brucei rhodesiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/patogenicidade , Tripanossomíase Africana/parasitologiaRESUMO
Target-based approaches for human African trypanosomiasis (HAT) and related parasites can be a valuable route for drug discovery for these diseases. However, care needs to be taken in selection of both the actual drug target and the chemical matter that is developed. In this article, potential criteria to aid target selection are described. Then the physiochemical properties of typical oral drugs are discussed and compared to those of known anti-parasitics.
Assuntos
Sistemas de Liberação de Medicamentos , Descoberta de Drogas , Tripanossomicidas/farmacocinética , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei rhodesiense/efeitos dos fármacos , Tripanossomíase Africana/tratamento farmacológico , Disponibilidade Biológica , Formas de Dosagem , Vias de Administração de Medicamentos , Esquema de Medicação , Resistência a Medicamentos , Humanos , Relação Estrutura-Atividade , Tripanossomicidas/química , Tripanossomicidas/farmacologia , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei gambiense/metabolismo , Trypanosoma brucei rhodesiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/metabolismo , Tripanossomíase Africana/parasitologiaRESUMO
SUMMARY This review presents a progression strategy for the discovery of new anti-parasitic drugs that uses in vitro susceptibility, time-kill and reversibility measures to define the therapeutically relevant exposure required in target tissues of animal infection models. The strategy is exemplified by the discovery of SCYX-7158 as a potential oral treatment for stage 2 (CNS) Human African Trypanosomiasis (HAT). A critique of current treatments for stage 2 HAT is included to provide context for the challenges of achieving target tissue disposition and the need for establishing pharmacokinetic-pharmacodynamic (PK-PD) measures early in the discovery paradigm. The strategy comprises 3 stages. Initially, compounds demonstrating promising in vitro activity and selectivity for the target organism over mammalian cells are advanced to in vitro metabolic stability, barrier permeability and tissue binding assays to establish that they will likely achieve and maintain therapeutic concentrations during in-life efficacy studies. Secondly, in vitro time-kill and reversibility kinetics are employed to correlate exposure (based on unbound concentrations) with in vitro activity, and to identify pharmacodynamic measures that would best predict efficacy. Lastly, this information is used to design dosing regimens for pivotal pharmacokinetic-pharmacodyamic studies in animal infection models.
Assuntos
Benzamidas/farmacocinética , Compostos de Boro/farmacocinética , Tripanossomicidas/farmacocinética , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei rhodesiense/efeitos dos fármacos , Tripanossomíase Africana/tratamento farmacológico , Administração Oral , Animais , Área Sob a Curva , Benzamidas/administração & dosagem , Benzamidas/sangue , Bioensaio , Barreira Hematoencefálica/efeitos dos fármacos , Compostos de Boro/administração & dosagem , Compostos de Boro/sangue , Permeabilidade Capilar , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Humanos , Concentração Inibidora 50 , Tripanossomicidas/administração & dosagem , Tripanossomicidas/sangue , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/crescimento & desenvolvimento , Tripanossomíase Africana/sangue , Tripanossomíase Africana/parasitologiaRESUMO
Human African trypanosomiasis (HAT, also called sleeping sickness), a neglected tropical disease endemic to sub-Saharan Africa, is caused by the parasites Trypanosoma brucei gambiense and T. brucei rhodesiense. Current drugs against this disease have significant limitations, including toxicity, increasing resistance, and/or a complicated parenteral treatment regimen. DB829 is a novel aza-diamidine that demonstrated excellent efficacy in mice infected with T. b. rhodesiense or T. b. brucei parasites. The current study examined the pharmacokinetics, in vitro and in vivo activity against T. b. gambiense, and time of drug action of DB829 in comparison to pentamidine. DB829 showed outstanding in vivo efficacy in mice infected with parasites of T. b. gambiense strains, despite having higher in vitro 50% inhibitory concentrations (IC50s) than against T. b. rhodesiense strain STIB900. A single dose of DB829 administered intraperitoneally (5 mg/kg of body weight) cured all mice infected with different T. b. gambiense strains. No cross-resistance was observed between DB829 and pentamidine in T. b. gambiense strains isolated from melarsoprol-refractory patients. Compared to pentamidine, DB829 showed a greater systemic exposure when administered intraperitoneally, partially contributing to its improved efficacy. Isothermal microcalorimetry and in vivo time-to-kill studies revealed that DB829 is a slower-acting trypanocidal compound than pentamidine. A single dose of DB829 (20 mg/kg) administered intraperitoneally clears parasites from mouse blood within 2 to 5 days. In summary, DB829 is a promising preclinical candidate for the treatment of first- and second-stage HAT caused by both Trypanosoma brucei subspecies.
Assuntos
Amidinas/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei rhodesiense/efeitos dos fármacos , Tripanossomíase Africana/tratamento farmacológico , Amidinas/farmacocinética , Animais , Esquema de Medicação , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Injeções Intraperitoneais , Melarsoprol/farmacologia , Camundongos , Testes de Sensibilidade Parasitária , Pentamidina/farmacologia , Fatores de Tempo , Tripanossomicidas/farmacocinética , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/crescimento & desenvolvimento , Tripanossomíase Africana/parasitologiaRESUMO
Trypanosoma brucei (T.b.) gambiense causes the chronic form of human African trypanosomiasis or sleeping sickness. One of the major problems with studying T.b. gambiense is the difficulty to isolate it from its original host and the difficult adaptation to in vivo and in vitro mass propagation. The objective of this study was to evaluate if an established method for axenic culture of pleomorphic bloodstream form T.b. brucei strains, based on methylcellulose containing HMI-9 medium, also facilitated the continuous in vitro propagation of other bloodstream form Trypanozoon strains, in particular of T.b. gambiense. Bloodstream form trypanosomes from one T.b. brucei, two T.b. rhodesiense, one T. evansi and seven T.b. gambiense strains were isolated from mouse blood and each was concurrently cultivated in liquid and methylcellulose-containing HMI-9 based medium, either with or without additional human serum supplementation, for over 10 consecutive sub passages. Although HMI-9 based medium supplemented with 1.1% (w/v) methylcellulose supported the continuous cultivation of all non-gambiense strains better than liquid media could, the in vitro cultivation of all gambiense strains was only achieved in HMI-9 based medium containing 1.1% (w/v) methylcellulose, 15% (v/v) fetal calf serum and 5% (v/v) heat-inactivated human serum.
Assuntos
Meios de Cultura/química , Metilcelulose , Soro , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Tripanossomíase Africana/parasitologia , Animais , Feminino , Congelamento , Humanos , Camundongos , Trypanosoma brucei gambiense/classificação , Trypanosoma brucei gambiense/fisiologiaRESUMO
The human parasite Trypanosoma brucei gambiense grew continuously at 37 degrees C in primary cultures of murine bone marrow. Cultured parasites remained virulent for mice. Rapid parasite growth coincided with the appearance of adherent adipocyte-epitheloid cell aggregates that also promoted hematopoiesis. This culture system should permit studies of host cell control of trypanosome proliferation, pathogenic effects of trypanosomes on blood cell development, and the relative trypanocidal and marrow suppressive activities of drugs.
Assuntos
Medula Óssea , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Animais , Células Cultivadas , Meios de Cultura , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Trypanosoma brucei brucei/crescimento & desenvolvimento , Tripanossomíase Africana/parasitologiaRESUMO
Human African trypanosomiasis is characterised by an important clinical diversity. Although Trypanosoma brucei gambiense field stocks isolated from patients in the same focus did not exhibit apparent genetic variability, they showed marked differences in terms of virulence (capacity to multiply inside a host) and pathogenicity (ability of producing mortality) in experimental murine infections. Two strains exhibiting opposite pathogenic and virulence properties in mouse were further investigated through their host-parasite interactions. In vitro, parasite bloodstream forms or soluble factors (or secretome) from both strains induced macrophage arginase as a function of their virulence. Arginase expression, a hallmark of macrophage alternative activation pathway, favours trypanosome bloodstream forms development. Moreover, a comparative proteomic study of the trypanosome stocks' secretomes evidenced both a differential expression of common molecules and the existence of stock specific molecules. This highlighted the potential involvement of the differential expression of the same genome in the diverse infectious properties of trypanosomes.
Assuntos
Trypanosoma brucei gambiense/imunologia , Trypanosoma brucei gambiense/patogenicidade , Animais , Arginase/biossíntese , Eletroforese em Gel Bidimensional , Feminino , Interações Hospedeiro-Parasita , Humanos , Macrófagos/enzimologia , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Parasitemia , Proteoma/análise , Proteínas de Protozoários/biossíntese , Análise de Sobrevida , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei gambiense/isolamento & purificação , Tripanossomíase Africana/parasitologia , VirulênciaRESUMO
BACKGROUND: Existing data on human African trypanosomiasis (HAT) due to Trypanosoma brucei gambiense among children are limited. Here, we described the demographic, clinical, diagnostic, treatment and outcome characteristics of HAT in pre-school children from Kajo-Keji County, South Sudan in comparison with older patients. METHODS: We did a retrospective analysis of HAT patients treated at the Kiri Sleeping Sickness Treatment Centre (SSTC), Kajo-Keji County, from June 2000 to December 2002. RESULTS: Of 1958 HAT patients, 119 (6.1%) were pre-school children (<6 years) including 56 (47%) in first-stage illness and 63 (53%) in second-stage. The proportion of children in second-stage HAT was significantly higher in very young children (<2 years). Walking and speech disturbances were more frequent in second-stage HAT but other neurological symptoms and signs were not associated with disease stage. Pentamidine treatment for first-stage illness was very safe and effective among pre-school children. In contrast, 4.9% of pre-school children in second-stage illness died during melarsoprol treatment and 46% had > or = 1 severe adverse event(s). Macular rash, jaundice and skin necrosis on injection site were significantly more frequent in this age group (p<0.05). Melarsoprol-induced encephalopatic syndrome was less frequent but more severe than in older age groups. CONCLUSION: The clinical features of T. b. gambiense HAT among pre-school children are insufficiently stage-specific. Therefore, laboratory-based staging is mandatory to prevent unnecessary harm to HAT patients caused by the high toxicity of melarsoprol.
Assuntos
Melarsoprol/administração & dosagem , Pentamidina/administração & dosagem , Tripanossomicidas/administração & dosagem , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Tripanossomíase Africana/tratamento farmacológico , Adolescente , Fatores Etários , Animais , Criança , Pré-Escolar , Humanos , Lactente , Infusões Intravenosas , Injeções Intramusculares , Estudos Retrospectivos , Sudão , Resultado do Tratamento , Tripanossomíase Africana/parasitologiaRESUMO
Diamidine-containing compounds have a long history of use in the treatment of African trypanosomiasis and leishmaniasis. The discovery that diamidine prodrugs possess in vivo antimicrobial activity when administered orally has led to a renewed interest in this class of compounds for the treatment of parasitic infections. In this review, the selectivity of diamidines against trypanosomes, Leishmania and Plasmodium is rationalized through mechanism-of-action studies. An overview of the antiprotozoal activities of newer diamidines and diamidine prodrugs is also presented, along with a summary of the progress made toward the clinical development of new diamidines for use against these parasitic diseases.
Assuntos
Antimaláricos/farmacologia , Antiprotozoários/farmacologia , Benzamidinas/metabolismo , Pentamidina/farmacologia , Pró-Fármacos/farmacologia , Tripanossomicidas/farmacologia , Animais , Antimaláricos/uso terapêutico , Antiprotozoários/uso terapêutico , Benzamidinas/farmacologia , Benzamidinas/uso terapêutico , Doença de Chagas/tratamento farmacológico , Ensaios Clínicos como Assunto , Avaliação Pré-Clínica de Medicamentos , Humanos , Leishmania/efeitos dos fármacos , Leishmania/crescimento & desenvolvimento , Leishmaniose/tratamento farmacológico , Malária Falciparum/tratamento farmacológico , Pentamidina/análogos & derivados , Pentamidina/uso terapêutico , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/crescimento & desenvolvimento , Pró-Fármacos/uso terapêutico , Tripanossomicidas/uso terapêutico , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/crescimento & desenvolvimento , Tripanossomíase Africana/tratamento farmacológicoRESUMO
Human African trypanosomiasis (HAT), due to the transmission of Trypanosoma brucei (T. b.) gambiense and T. b. rhodesiense by tsetse flies, is re-emerging in inter-tropical Africa. It evolves from the hemolymphatic Stage I to the meningo-encephalitic Stage II. The latter is generally treated with melarsoprol, an arseniate provoking often a deadly encephalopathy. A precise determination of the HAT evolution stage is therefore crucial. Stage II patients show: (i) a deregulation of the 24-h distribution of the sleep-wake alternation; (ii) an alteration of the sleep structure, with frequent sleep onset rapid eye movement (REM) periods (SOREMPs). Gambian HAT was diagnosed in eight patients (four, Stage II; three, Stage I; one, "intermediate" case) at the trypanosomiasis clinic at Viana (Angola). Continuous 48-h polysomnography was recorded on Oxford Medilog 9000-II portable systems before and after treatment with melarsoprol (Stage II) or pentamidine (Stage I and "intermediate" stage). Sleep traces were visually analyzed in 20-s epochs using the PRANA software. Stage II patients showed the complete sleep-wake syndrome, partly reversed by melarsoprol 1 month later. Two Stage I patients did not experience any of these alterations. However, the "intermediate" and one Stage I patients exhibited sleep disruptions and/or SOREMPs, persistent after pentamidine treatment. Polysomnography may represent a diagnostic tool to distinguish the two stages of HAT. Especially, SOREMPs appear shortly after the central nervous system invasion by trypanosomes. The reversibility of the sleep-wake cycle and sleep structure alterations after appropriate treatment constitutes the basis of an evaluation of the healing process.
Assuntos
Polissonografia , Sono , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Tripanossomíase Africana/diagnóstico , Adolescente , Adulto , Animais , Criança , Feminino , Humanos , Masculino , Melarsoprol/uso terapêutico , Tripanossomicidas/uso terapêutico , Tripanossomíase Africana/tratamento farmacológico , Tripanossomíase Africana/fisiopatologiaRESUMO
The existence of a pig reservoir for human African trypanosomosis (HAT) due to Trypanosoma brucei gambiense complicates the fight against this disease. This study, reports results obtained from pigs, which were inoculated with the blood of a person, suffering from HAT in Cameroon. The pigs were reared and kept in the shelter from all contact with Glossina, and monitored for 188 days. The seroconversion was checked by agglutination assays for trypanosomosis (CATT 1.3 and LATEX/T.b.gambiense). The parasitemia was measured by quantitative buffy coat method (QBC) and by polymerase chain reaction method (PCR). In addition, growth was recorded as well as blood counting and blood formulas. The results showed that the pigs were trypanotolerant and cure themselves in less than 6 months. It is concluded that sterilisation of this reservoir could be achieved by tsetse-control measures in 1 year. It confirms the strategy to complement screening and treatment of HAT with tsetse fly control measures.
Assuntos
Portador Sadio/veterinária , Reservatórios de Doenças/veterinária , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/parasitologia , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Tripanossomíase Africana/prevenção & controle , Tripanossomíase Africana/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Portador Sadio/parasitologia , Feminino , Controle de Insetos , Insetos Vetores/parasitologia , Testes de Fixação do Látex/veterinária , Masculino , Parasitemia/veterinária , Suínos , Tripanossomíase Africana/parasitologia , Moscas Tsé-Tsé/parasitologiaRESUMO
We examined whether heparin administration influences in vivo trypanosome proliferation in infected rats. Administration of heparin every 8 hr via cardiac catheter inhibited growth of Trypanosoma brucei gambiense and prolonged survival of treated rats. Heparin administration increased lipoprotein lipase activity, high-density lipoprotein (HDL) concentration in the blood, and haptoglobin messenger RNA content of the liver. The presence of heparin in culture media did not directly affect proliferation of trypanosomes in vitro. However, the addition of plasma from infected rats treated with heparin to culture media decreased the number of trypanosomes. This effect was decreased by incubating the trypanosomes with benzyl alcohol, a known inhibitor of receptor-mediated endocytosis of lipoprotein. These data suggested that heparin administration reduced the number of trypanosomes in infected rats. Trypanosome lytic factor, a HDL and haptoglobin-related protein, protects humans and some animals from infection by Trypanosoma brucei brucei. In rats, increases in HDL and haptoglobin may affect the proliferation of T. b. gambiense.
Assuntos
Anticoagulantes/farmacologia , Heparina/farmacologia , Trypanosoma brucei gambiense/efeitos dos fármacos , Tripanossomíase Africana/tratamento farmacológico , Animais , Anticoagulantes/uso terapêutico , HDL-Colesterol/sangue , HDL-Colesterol/efeitos dos fármacos , Haptoglobinas/efeitos dos fármacos , Haptoglobinas/genética , Haptoglobinas/metabolismo , Heparina/uso terapêutico , Lipase Lipoproteica/sangue , Lipase Lipoproteica/efeitos dos fármacos , Fígado/química , Fígado/efeitos dos fármacos , Masculino , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Tripanossomíase Africana/parasitologiaRESUMO
The problem of tsetse-transmitted trypanosomiasis occurs only in sub-Saharan Africa, where it represents a major constraint to socio-economic development. The East African form of sleeping sickness, caused by Trypanosoma brucei rhodensiense, is an acute and fatal disease, whereas the West African form, caused by Trypanosoma brucei gambiense, is generally more chronic and debilitating. The African governments have developed a new initiative, known as the Pan African Tsetse and Trypanosomiasis Eradication Campaign, which seeks to employ an area-wide approach and appropriate fly suppression methods to eradicate tsetse from areas of tsetse infestation, at a time, to ultimately create tsetse-free zones.
Assuntos
Controle de Insetos/métodos , Insetos Vetores , Tripanossomíase/prevenção & controle , Moscas Tsé-Tsé , África Subsaariana , Animais , Bovinos , Feminino , Humanos , Masculino , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei rhodesiense/crescimento & desenvolvimentoRESUMO
High resolution two-dimensional polyacrylamide gel (2D gel) electrophoresis and autoradiography were used to analyze the protein gene products of African trypanosomes biosynthetically labelled with [35S]methionine. Using cloned populations of parasites it was found that: antigenically different bloodstream trypanosomes from the same serodeme differed only in their variant surface glycoproteins; Trypanosoma brucei, T.b. rhodesiense and T.b. gambiense subspecies could be distinguished on the basis of differences in expressed proteins; transformation from bloodstream trypomastigotes to procyclic epimastigote culture forms was accompanied by loss of variant surface glycoproteins and several other qualitative and quantitative changes in minor proteins. The results indicate that 2D gel analysis may allow improved classification of African trypanosomes (based on the observation of hundreds of protein markers) and may also provide a general technique for the identification of lifecycle stage specific markers.
Assuntos
Antígenos de Superfície/genética , Trypanosoma brucei brucei/crescimento & desenvolvimento , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Animais , Eletroforese em Gel de Poliacrilamida , Genes , Metionina/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Fenótipo , Especificidade da Espécie , Radioisótopos de Enxofre , Trypanosoma brucei brucei/imunologia , Trypanosoma brucei gambiense/imunologiaRESUMO
We have used pulse-chase immunoprecipitations methods to study early post-translational processing of CBI-gp, a lysosomal membrane glycoprotein expressed by African trypanosomes, Rap67, a polyclonal antibody to CBI-gp, immunoprecipitated a 100-kDa glycoprotein, gp100, from both bloodstream forms (BF) and procyclic forms (PF) of Trypanosoma brucei gambiense immediately after a 5-min pulse with radiomethionine. N-Glycanase digestion released a 67-kDa core protein, p67, from gp100 of both life cycle forms V8 protease digestion of p67 from BF and PF yielded 13 identical methionyl peptides, suggesting that gp100 from both life cycle forms have very similar or identical p67 core molecules. In BF, gp 100 carried both endoglycosidase H (EndoH)-resistant and EndoH-sensitive, N-linked oligosaccharides immediately after labeling. In PF, all the N-linked sugars on gp100 were EndoH sensitive. In BF, gp100 chased progressively into slower migrating 150-180-kDa components that obtained the CBI epitope, traveled to the cell surface where they could be biotinylated, and were proteolytically processed. The increase in mass of gp100 during chase in BF resulted from an elongation of N-linked oligosaccharides. Maturation of gp100 into 150-180-kDa CBI-gp was inhibited if BF were chased in the presence of glucosidase inhibitors castanospermine or deoxynojirimycin. In PF, gp100 did not increase in mass, could not be biotinylated on the cell surface, and was not proetolyzed during extended chases. Cryoimmunoelectron microscopy revealed that the antigens detected by rap67 are abundant in lysosomes and endosomes in both BF and PF. Thus, BF and PF express very similar or identical lysosomal membrane glycoproteins but process and transport them in very different ways.
Assuntos
Glicoproteínas de Membrana/metabolismo , Proteínas de Protozoários/metabolismo , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Trypanosoma brucei gambiense/metabolismo , Animais , Anticorpos Monoclonais , Antígenos de Protozoários/metabolismo , Transporte Biológico Ativo , Biotina , Glicosilação , Lisossomos/imunologia , Lisossomos/metabolismo , Lisossomos/ultraestrutura , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Imunoeletrônica , Processamento de Proteína Pós-Traducional , Proteínas de Protozoários/imunologia , Trypanosoma brucei gambiense/ultraestruturaRESUMO
Trypanosoma brucei gambiense and T. b. rhodesiense are protozoan parasites causing sleeping sickness in humans due to their resistance to lysis by normal human serum (NHS). Based on the observation that the resistance gene of T. b. rhodesiense encodes a truncated form of the variant specific glycoprotein (VSG), we cloned a similar gene in T. b. gambiense using reverse transcription-linked polymerase chain reaction with VSG-specific primers. This gene, termed TgsGP for T. gambiense-specific glycoprotein, was found to be specific to T. b. gambiense. It is located close to a telomere and is transcribed by a pol II RNA polymerase, only at the bloodstream stage of the parasite development. TgsGP encodes a 47-kDa protein consisting of a N-terminal VSG domain presumably provided with a glycosylphosphatidylinositol (GPI) anchor sequence, similar to the pESAG6 subunit of the trypanosomal transferrin receptor. TgsGP is located in the flagellar pocket, and contains the linear N-linked polyacetyllactosamine characteristic of the endocytotic machinery of T. brucei. These observations strongly suggest that TgsGP is a T. b. gambiense specific receptor. Since stable expression of this protein in T. b. brucei did not confer resistance to NHS, TgsGP may either need another factor to achieve this purpose or fulfils another function linked to adaptation of the parasite to man.
Assuntos
Antígenos de Protozoários/genética , Proteínas , Receptores da Transferrina/genética , Trypanosoma brucei gambiense/imunologia , Glicoproteínas Variantes de Superfície de Trypanosoma/genética , Sequência de Aminoácidos , Animais , Antígenos de Protozoários/isolamento & purificação , Antígenos de Protozoários/metabolismo , Northern Blotting , Resistência a Medicamentos/genética , Genes de Protozoários , Glicosilfosfatidilinositóis/metabolismo , Humanos , Dados de Sequência Molecular , Peso Molecular , Proteínas de Protozoários , RNA Polimerase II/metabolismo , RNA Mensageiro/análise , RNA de Protozoário/genética , Receptores da Transferrina/isolamento & purificação , Receptores da Transferrina/metabolismo , Sensibilidade e Especificidade , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Telômero/genética , Trypanosoma brucei gambiense/citologia , Trypanosoma brucei gambiense/efeitos dos fármacos , Trypanosoma brucei gambiense/crescimento & desenvolvimento , Glicoproteínas Variantes de Superfície de Trypanosoma/isolamento & purificação , Glicoproteínas Variantes de Superfície de Trypanosoma/metabolismoRESUMO
Small-molecule oxidosqualene cyclase (OSC) inhibitors were found to be effective in assays against cloned OSC-like enzymes from human pathogens. A combinatorial library was prepared and used to identify lead compounds that inhibit the growth of Trypanosoma cruzi, Leishmania mexicana amazonensis, and Pneumocystis carinii in culture. Selectivity for the microorganisms in preference to mammalian cells was observed.