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Dietmar Steverding

Publications and source records attributed to Dietmar Steverding.

At least 19 recordsLinked to original sources
PubMed2006

On the significance of host antibody response to the Trypanosoma brucei transferrin receptor during chronic infection.

The transferrin (Tf) receptor of Trypanosoma brucei (TbTfR) is encoded by two expression-site-associated genes, ESAG6 and ESAG7. There are around 20 different expression sites containing different copies of these genes that encode TbTfRs with quite distinct affinities for Tf of various hosts. It was proposed that T. brucei has developed multiple expression sites encoding different TbTfRs to ensure sufficient iron uptake in the presence of antibodies competing for binding to Tf. Here it is shown that anti-TbTfR antibody titres produced during chronic murine trypanosomiasis are only one-tenth of those achieved by immunisation of mice using recombinant TbTfR. Calculations indicate that the concentrations of competing anti-TbTfR antibodies present during chronic T. brucei infection are too low to deprive the parasite of iron. In addition, during human African trypanosomiasis the antibody response to the TbTfR seems to be poor and transient. Altogether, the results suggest that the host antibody response to the TbTfR during chronic infection with T. brucei is too low, if present at all, to prevent sufficient iron uptake by bloodstream forms to promote their growth.

Animals↗
PubMed2006

In vitro growth inhibition of bloodstream forms of Trypanosoma brucei and Trypanosoma congolense by iron chelators.

African trypanosomes exert significant morbidity and mortality in man and livestock. Only a few drugs are available for the treatment of trypanosome infections and therefore, the development of new anti-trypanosomal agents is required. Previously it has been shown that bloodstream-form trypanosomes are sensitive to the iron chelator deferoxamine. In this study the effect of 13 iron chelators on the growth of Trypanosoma brucei, T. congolense and human HL-60 cells was tested in vitro. With the exception of 2 compounds, all chelators exhibited anti-trypanosomal activities, with 50% inhibitory concentration (IC50) values ranging between 2.1-220 microM. However, the iron chelators also displayed cytotoxicity towards human HL-60 cells and therefore, only less favourable selectivity indices compared to commercially available drugs. Interfering with iron metabolism may be a new strategy in the treatment of trypanosome infections. More specifically, lipophilic iron-chelating agents may serve as lead compounds for novel anti-trypanosomal drug development.

Journal Article↗
PubMed2006

Evaluation of the anti-trypanosomal activity of tyropeptin A.

The natural compound tyropeptin A, a new peptidyl aldehyde proteasome inhibitor, was tested for its trypanocidal activity in vitro using culture-adapted bloodstream forms of Trypanosoma brucei. The concentrations of tyropeptin A required to reduce the growth rate by 50 % and to kill all cells were 10 and 100 times lower for bloodstream-form trypanosomes than for human leukaemia HL-60 cells, respectively. Enzymatic analysis showed that the trypsin-like activity of the trypanosome proteasome and the chymotrypsin-like activity of the mammalian proteasome are particularly sensitive to inhibition by tyropeptin A. The results suggest that natural compounds targeting the trypsin-like activity of the proteasome may serve as leads for rational drug development of novel anti-trypanosomal agents.

Animals↗
PubMed2006

A new initiative for the development of new diagnostic tests for human African trypanosomiasis.

Human African trypanosomiasis is a threat to millions of people living in sub-Saharan countries and is fatal unless treated. At present, the serological and parasitological tests used in the field for diagnosis of sleeping sickness have low specificity and sensitivity. There is clearly an urgent need for accurate tools for both diagnosis and staging of the disease. The Foundation for Innovative New Diagnostics and the World Health Organization have announced that they will collaborate to develop and evaluate new diagnostic tests for human African trypanosomiasis.

Journal Article↗
PubMed2006

Cysteine proteinase inhibitors as therapy for parasitic diseases: advances in inhibitor design.

Clan CA (papain-like) cysteine proteinases of protozoan parasites are validated targets for the rational design of new anti-parasitic chemotherapies. Peptidyl and peptidomimetic proteinase inhibitors of differing chemistries limit parasite survival in vitro and in vivo. Also, the development of activity-based affinity labels has enabled the identification and characterization of potential cysteine proteinase targets in situ. This article reviews the biology and physicochemistry of parasite proteinases and the ongoing design of peptidyl and non-peptidyl inhibitors to generate anti-parasitic compounds of greater efficacy with decreased toxicity to the host.

Cysteine Proteinase Inhibitors↗
PubMed2005

Effect of Australian tea tree oil on Gyrodactylus spp. infection of the three-spined stickleback Gasterosteus aculeatus.

Gyrodactylus spp. infections of commercially farmed fishes are responsible for significant economic losses. Existing treatments have proved uneconomic, stressful to the fishes, and ecologically damaging. Essential oils are naturally occurring compounds that exhibit a wide range of anti-microbial and anti-fungal activities. This study explored the possibility of using Australian tea tree (Melaleuca alternifolia) oil (TTO) to treat Gyrodactylus spp. infection on the three-spined stickleback Gasterosteus aculeatus. In the presence of 0.01 % Tween 80 as an emulsifier, TTO treatments at concentrations between 3 and 30 ppmv (parts per million by volume) lowered the prevalence and significantly reduced the parasite burden of sticklebacks naturally infected with Gyrodactylus spp. In addition, Tween 80 alone exhibited parasiticidal activity against Gyrodactylus spp. These findings show the potential of TTO in combination with Tween 80 as an effective treatment of Gyrodactylus spp. infection of fishes.

Animals↗
PubMed2005

Intracellular positioning of isoforms explains an unusually large adenylate kinase gene family in the parasite Trypanosoma brucei.

Adenylate kinases occur classically as cytoplasmic and mitochondrial enzymes, but the expression of seven adenylate kinases in the flagellated protozoan parasite Trypanosoma brucei (order, Kinetoplastida; family, Trypanosomatidae) easily exceeds the number of isoforms previously observed within a single cell and raises questions as to their location and function. We show that a requirement to target adenylate kinase into glycosomes, which are unique kinetoplastid-specific microbodies of the peroxisome class in which many reactions of carbohydrate metabolism are compartmentalized, and two different flagellar structures as well as cytoplasm and mitochondrion explains the expansion of this gene family in trypanosomes. The three isoforms that are selectively built into either the flagellar axoneme or the extra-axonemal paraflagellar rod, which is essential for motility, all contain long N-terminal extensions. Biochemical analysis of the only short form trypanosome adenylate kinase revealed that this enzyme catalyzes phosphotransfer of gamma-phosphate from ATP to AMP, CMP, and UMP acceptors; its high activity and specificity toward CMP is likely to reflect an adaptation to very low intracellular cytidine nucleotide pools. Analysis of some of the phosphotransfer network using RNA interference suggests considerable complexity within the homeostasis of cellular energetics. The anchoring of specific adenylate kinases within two distinct flagellar structures provides a paradigm for metabolic organization and efficiency in other flagellates.

Adenylate Kinase↗
PubMed2005

Anti-trypanosomal activities of DNA topoisomerase inhibitors.

Only four drugs are available for chemotherapy of human African sleeping sickness with undesirable toxic side effects. The development of new anti-trypanosomal drugs is, therefore, urgently required. In this study, 15 DNA topoisomerase inhibitors, including approved anti-cancer drugs, were tested for in vitro activity against bloodstream forms of Trypanosoma brucei and human leukaemia HL-60 cells. All compounds exhibited anti-trypanosomal activity, with ED50 values ranging between 3 nM and 30 microM, and MIC values between 100 nM and >100 microM. The trypanocidal activities of the most effective DNA topoisomerase inhibitors, aclarubicin, doxorubicin and mitoxantrone, were comparable with those of commercial anti-trypanosomal drugs. These data support the use of DNA topoisomerase inhibitors as lead compounds for anti-trypanosomal drug development.

Animals↗
PubMed2005

Novel antitrypanosomal agents.

Trypanosomes are the causative agents of Chagas' disease in Central and South America and sleeping sickness in sub-Saharan Africa. The current chemotherapy of the human trypanosomiases relies on only six drugs, five of which were developed > 30 years ago. In addition, these drugs display undesirable toxic side effects and the emergence of drug-resistant trypanosomes has been reported. Therefore, the development of new drugs in the treatment of Chagas' disease and sleeping sickness is urgently required. This article summarises the recent progress in identifying novel lead compounds for antitrypanosomal chemotherapy. Particular emphasis is placed on those agents showing promising, selective antitrypanosomal activity.

Animals↗
PubMed2004

Trypanocidal activities of trileucine methyl vinyl sulfone proteasome inhibitors.

Previous studies have shown that proteasome inhibitors are novel agents for chemotherapy of human African trypanosomiasis or sleeping sickness. In this study, five peptide trileucine methyl vinyl sulfones with different N-terminal substituents were tested for their trypanocidal activities in vitro using culture-adapted bloodstream forms of Trypanosoma brucei. Two inhibitors displayed promising anti-trypanosomal activities with ED50 values in the sub-micromolar range. Higher trypanocidal activity of the compounds generally corresponded to a higher k(obs)/[I] value for inhibition of the trypsin-like activity but not for the inhibition of the chymotrypsin-like activity of the proteasome. These data suggest that inhibitors with strong activity against the trypsin-like activity of the proteasome are the rational choice for future anti-sleeping sickness drug development.

Animals↗
PubMed2004

On the role of blue shadows in the visual behaviour of tsetse flies.

Tsetse flies (Glossina spp.), the vector for African trypanosomiasis, are highly attracted by blue and black surfaces. This phototactic behaviour has long been exploited to trap tsetse flies as one measure in the control of African trypanosomiasis. However, why blue and black are so attractive for tsetse flies is still unknown. We propose that the combination of blue and black is attractive for many Glossina species because when searching for a shady resting place to pass the day, the flies are probably guided by the blueness and darkness of daytime shadows. In contrast to people's experience that daytime shadows are colourless, actually on a sunny day all shadows are tinted bluish by the scattered blue skylight.

Animals↗
PubMed2004

Real-time PCR for detection of Trypanosoma brucei in human blood samples.

We have developed a real-time PCR assay for detection of Trypanosoma brucei DNA in human blood samples. The PCR was conducted with newly designed primers targeting the 177-bp repeat satellite DNA in T. brucei and with Sybr Green to monitor the amplicon accumulation. DNA purification using Chelex 100 resin was performed on blood samples collected on Whatman FTA cards and was shown to be a simple and quantitative method as revealed by real-time PCR. The detection limit of the assay was 100 trypanosomes per mL blood, corresponding to an analytical sensitivity of 0.1 genome equivalents. Trypanosome DNA was detected in all blood samples from sleeping sickness patients and, furthermore, the identity of the amplicon was confirmed in all assays by dissociation analysis. Although template DNA from blood samples was amplified with significantly lower efficiency than genomic DNA, similar efficiency between all assays ensured quantitative results. No amplicon product was obtained with samples from uninfected individuals. The results indicate that the real-time PCR assay described is a rapid and sensitive method suitable for the detection of T. brucei in human blood samples in routine clinical laboratory practice.

Animals↗
PubMed2004

Trypanocidal effect of alpha',beta'-epoxyketones indicates that trypanosomes are particularly sensitive to inhibitors of proteasome trypsin-like activity.

Previous studies have shown that the proteasome of Trypanosoma brucei is a candidate for novel chemotherapy of African sleeping sickness. In this study, two potent and highly selective alpha',beta'-epoxyketones peptide proteasome inhibitors, epoxomicin and YU101, have been tested for their trypanocidal activities in vitro using culture-adapted bloodstream forms of T. brucei. Both inhibitors displayed promising anti-trypanosomal activities with ED(50) and ED(90) values in the low to mid nanomolar range. Based on MIC values, epoxomicin exhibited a selectivity index approaching those of commercially available drugs. Enzymatic analyses of proteasomal peptidase activities revealed that, compared with mammalian cells, trypanosomes are particular sensitive to inhibition of the trypsin-like activity of the proteasome. In conclusion, the data suggests that proteasome inhibitors targeting the trypsin-like activity are the rational choice for future anti-trypanosomal drug development.

Animals↗
PubMed2004

Trypanosoma brucei: unexpected azide sensitivity of bloodstream forms.

Bloodstream forms of Trypanosoma brucei lack cytochromes and are, therefore, insensitive to cyanide. Azide is a toxic anion that bears chemical and biological properties in common with cyanide and may act in a similar way by inhibition of cytochrome c oxidase. It was, therefore, surprising to find that bloodstream forms of T. brucei are sensitive to azide; growth is reduced by 50% with 0.1 mM azide. So far, the only enzyme known in bloodstream forms of T. brucei to be sensitive to azide is the iron-containing superoxide dismutase. However, because the activity of the superoxide dismutase was not affected in parasites incubated for 16 hr with 0.5 mM azide (a concentration at which no cell proliferates), the toxic action of azide cannot be due to inhibition of this enzyme. These results indicate that the general toxicity of azide is different from that of cyanide.

Animals↗
PubMed2003

Improved trypanocidal activities of cathepsin L inhibitors.

The major lysosomal cysteine proteinase of African trypanosomes is a candidate target for novel chemotherapy of sleeping sickness. This cathepsin L-like enzyme is termed rhodesain and brucipain in Trypanosoma brucei rhodesiense and Trypanosoma brucei brucei, respectively. Three potent and selective dipeptidyl cathepsin L inhibitors have been investigated for their trypanocidal activities in vitro using culture-adapted bloodstream forms of T. b. brucei. Compared with general cysteine proteinase inhibitors used previously by ourselves and others, the present inhibitors had improved selectivity indices and, importantly, anti-trypanosomal activities comparable with those of commercial anti-sleeping sickness drugs. Using purified recombinant rhodesain, potent k(inact)/Ki values of up to 2.3x10(6) M(-1) s(-1) were recorded with the inhibitors. Also, all inhibitors blocked proteinolysis in the lysosome consistent with the inhibition of rhodesain/brucipain. In conclusion, the data support the potential of cathepsin L inhibitors for rational anti-trypanosomal drug development.

Animals↗
PubMed2003

The significance of transferrin receptor variation in Trypanosoma brucei.

The transferrin receptor of Trypanosoma brucei is encoded by genes located in different expression sites. The various expression sites encode slightly different transferrin receptors, which differ substantially in their affinity for transferrin of different host species. It was proposed that T. brucei has developed multiple expression sites encoding different transferrin receptors not only to cope with the diversity of mammalian transferrins, but also to ensure sufficient iron uptake in the presence of anti-transferrin receptor antibodies. This article shows that calculations based on K(d) values argue against the first part of the hypothesis, but might support the second part.

Animals↗
PubMed2002

Trypanosoma brucei: in vitro slender-to-stumpy differentiation of culture-adapted, monomorphic bloodstream forms.

Pleomorphic Trypanosoma brucei strains are characterized by their ability to differentiate from replicating long slender forms into non-dividing short stumpy forms in the mammalian host. The differentiation process can be efficiently induced in vitro by treatment with the membrane-permeable cAMP derivative 8-(4-chlorophenylthio)-cAMP (pCPTcAMP). In contrast, monomorphic T. brucei strains do not differentiate to stumpy forms in the host. Here, we show that exposure of monomorphic, culture-adapted T. brucei bloodstream forms to pCPTcAMP allowed their subsequent differentiation into short stumpy forms. The stumpy nature of pCPTcAMP-treated parasites was confirmed by (1) morphological change, (2) inhibition of growth and DNA synthesis, (3) cell cycle arrest in the G(1)/G(0) phase, (4) expression of NADH diaphorase activity and dihydrolipoamide dehydrogenase, (5) disappearance of the small subunit of ribonucleotide reductase, (6) up-regulation of the major lysosomal membrane protein, and (7) efficient transformation into replicating procyclic insect forms after induction with citrate/cis-aconitate. Our results indicate that the inability of monomorphic T. brucei bloodstream forms to differentiate into short stumpy forms in the host may be due to a failure in the signalling pathway rather than in the differentiation process itself. Treatment of monomorphic bloodstream trypanosomes with pCPTcAMP could be a useful method for identifying the genes involved in the slender-to-stumpy differentiation process.

Animals↗