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中国畜牧兽医学会家畜寄生虫学分会第九次学术研讨会

作者： Clare L．Allen David Goulding Wei-Lian Chung Mark C．Field Department of Pathology Tennis Court RoadUniversity of CambridgeCambridge CB2 1QPUK

In the divergent eukaryote Trypanosoma brucei the architecture of the major intracellular transport routes responsible for exocytosis, endocytosis and recycling are broadly similar to higher eukaryotes, but with a number of important differences. The system is exclusively clathrin-mediated and dynamin-independent, unlike metazoa and yeast where clathrin-independent mechanisms exist and clathrin-dependent endocytosis usually requires the function of dynamin. T. brucei also lacks a number of adaptor proteins that mediate recognition of cargo molecules trafficking through the endosomal system, including the membrane adaptin complex AP-2, which in most systems is responsible for recognition of peptide-based sorting signals present within the cytoplasmic regions of trans-menibrane domain surface proteins. Here we have determined the functions of the trypanosome AP-1 complex, which is responsible for trans-Golgi network and endosomal sorting in higher eukaryotes, and is the most closely related adaptor complex to AP-2. By RNAi mediated knockdown we demonstrate that the complex is essential, similar to Dictyostelium but differentiating trypanosomes from yeasts and metazoa. Further, we show that the ? 1 and ? -subunits cross suppress at the protein level indicating they are part of the same complex. By knockdown of the ? -subunit we find that there are pleiotropic defects to membrane transport, but that the major focus appears to be post-Golgi lysosomal transport as detected by delayed processing of late cleavage events of p67?a major lysosomal protein of trypanosomes. Further, by site-directed mutagenesis we demonstrate that p67 contains a cannonical AP-1 sorting signal that is required for lysosomal targeting. These data demonstrate a conservation of function for AP-1 in trypanosomes and higher eukaryotes and the evolutionary implications are discussed.

关键词： trypanosome protein transport vesicle trafficking endocytosis adaptin

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Antitrypanosomal potentials of the extract and fractions of Abrus precatorius seeds

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Asian Pacific Journal of Tropical Medicine 2012年第11期5卷 857-861页

作者： Nwodo NJ Nwodo OFC Department of Pharmaceutical Chemistry University of NigeriaNsukka Nigeria Department of Biochemistry University of NigeriaNsukka Nigeria

Objective:To evaluate the in vivo trypanocidal activity of the methanol extract and fractions of Abrus precatorius seeds in ***:Parasiteamia was induced unto mice by intraperitoneal injection of 1.25×105 Trypanosoma in normal *** days when a high level of parasiteamia was established treatment commenced until ten *** mice were treated with 10,20 and 40 mg/kg *** the extract and 5 and 10 mg/kg *** the fraction（F2）,respectively for 5 *** acelurate at the dose of 3.5 mg/kg *** two days was used as the reference *** level of parasitaemia and packed cell volume（PCV） of the animals estimated. Results:At doses of 10,20 and 40 mg/kg the crude extract showed a sharp reduction in the level of parasitaemia in mice compared with the untreated *** mice treated with F,at doses of 5 and 10 mg/kg showed a sharp reduction in the level of parasitamia to zero in day 9,and a gradual recovery from the 12th day of *** effect is comparable to that of the mice treated with 7 mg/kg of standard drug diminazene *** PCV of the treated showed a gradual decrease with time,but not as much as the untreated *** screening revealed the presence of glycosides,alkaloids,carbohydrates,tannins and proteins in the Abrus precatorius powder while F2 was rich in ***:This study shows that both the extract and the fractions of Abrus precatorius seeds exhibited a promising trypanocidal *** may be responsible for the observed activity.

关键词： Abrus precatorius Seed extract Trypanocidal effect Parasiteamia Fractions trypanosome brucei brucei Berenil

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In vitro and in vivo trypanocidal action of aescin and aescin liposomes against Trypanosoma evansi in experimental mice

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Asian Pacific Journal of Tropical Biomedicine 2014年第12期4卷 947-951页

作者： Matheus Dellamea Baldissera Nathieli Bianchin Bottari Thirssa Helena Grando Roberto Christ Vianna Santos Ana Julia Figueiro Dalcin Patrfcia Gomes Renata Platcheck Raffin Carine Eloise Prestes Zimmerman Janio Morais Santurio Silvia Gonzalez Monteiro Aleksandro Schafer Da Silva Department of Microbiology and Parasitology Universidade Federal de Santa Maria(UFSM) Laboratory of Microbiology Centro Universitdrio Franciscano Laboratory of Nanotechnology Centro Universitdrio Franciscano Department of Animal Science Universidade do Estado de Santa Catarina(UDESC)

Objective:To verify the trypanocidal effectiveness of aescin and aescin liposomes against Trypanosoma evansi in vitro and in ***:Aescin and aescin liposomes were used in vitro on trypomastigotes at different concentrations(0.5%,1.0%and 2.0%) and exposure times(0,1,3,6 and 9 h).In vivo tests were performed using mice as the experimental *** evansi infected mice were treated with aescin and aescin liposomes with doses of 60 and 100 mg/kg during 4 ***:The three concentrations tested in free form and nanoencapsulated showed trypanocidal activity in vitro,completely eliminating the parasites in small concentration after6 h of *** treated with aescin(100 mg/kg) and aescin liposomes(100 mg/kg)showed increase in longevity,however without curative ***:Active compounds present in natural products,such as aescin,may potentiate the treatment of trypanosomosis when used in association with other trypanocidal drugs.

关键词： Surra Nanotechnology Aesculus hippocastanum trypanosome

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Improved Detection of Sleeping Sickness Cases by LED Fluorescence Microscopy: Evidence from a Prospective Multi-Centric Study in the Democratic Republic of the Congo

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Microscopy Research 2015年第2期3卷 17-25页

作者： Patrick Mitashi Pascal Lutumba Crispin Lumbala Paul Bessell Sylvain Biéler Joseph Mathu Ndung’u Tropical Medicine Department Faculty of Medicine University of Kinshasa Kinshasa Democratic Republic of the Congo Programme National de Lutte contre la Trypanosomiase Humaine Africaine (PNLTHA) Croisement de l’Avenue de la Libération et du Boulevard Triomphal 1 Kinshasa Democratic Republic of the Congo Foundation for Innovative New Diagnostics (FIND) Campus Biotech Chemin des Mines 9 Geneva Switzerland

Background: Confirmatory diagnosis of Trypanosoma brucei gambiense human African trypanosomiasis (HAT) is based on demonstration of parasites by microscopy. However, the sensitivity of routine microscopy methods is very low, and many cases are missed and left untreated. A clinical study was conducted in the Democratic Republic of the Congo to evaluate the accuracy of improved microscopy methods in diagnosis of HAT. These included examination by fluorescence microscopy (FM) of acridine orange (AO) stained smears of whole blood and smears made following a new procedure for concentrating trypanosomes by selective lysis of red blood cells (RBC). Methodology/Principal Findings: Venous blood was collected from 213 HAT cases, 101 HAT suspects and 95 controls and used to determine the accuracy of four microscopy methods: bright field microscopy of Giemsa-stained thick blood smears, FM of AO-stained thick blood smears, FM of AO-stained thick blood smears prepared after RBC lysis and concentration, and FM of AO-stained thin blood smears prepared after RBC lysis and concentration. The sensitivity of FM using thick blood smears stained with AO was 3 times higher than bright field microscopy using Giemsa-stained thick blood smears [19.7% (95% CI: 14.9% - 25.6%) versus 6.1% (95% CI: 3.6% - 10.2%)]. When the RBC lysis and concentration procedure was included, sensitivity of the test was further enhanced to 23.0% (95% CI: 17.9% - 29.1%) with thick blood smears and 34.3% (95% CI: 28.2% - 40.9%) with thin blood smears. Specificity of all four microscopy methods was 100% (95% CI: 96.1% - 100.0%). However, the miniature anion exchange chromatography technique (mAECT) and capillary tube centrifugation (CTC) method remained more sensitive. Conclusions: These new methods have practical advantages, including shorter staining time, ease of demonstration of parasites, and the possibility of archiving slides. They could, therefore, be alternative methods to improve case detection where concen

关键词： Human African Trypanosomiasis trypanosome LED Fluorescence Microscopy Red Blood Cell Lysis Diagnosis

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