Avian malaria parasites are responsible for severe diseases in some domestic and wild birds. These parasites are cosmopolitan in distribution; they are widespread in Europe, including the Baltic region. A peculiarity of current studies of avian Plasmodium species is that information about ecology, distribution, prevalence and other aspects of their biology has been accumulated using free-living birds. To elucidate the significance of malaria infections and their impact on host fitness, behaviour, sexual selection and parasite-host co-evolution, experimental information about Plasmodium spp. virulence, specificity and dynamics of parasitemia in different avian hosts is crucial. Unfortunately such studies remain uncommon. Theobjective of this study was to obtain new field and laboratory experimental data about the biology of avian malaria parasites and to link PCR-based information with data from traditional parasitology. It was demonstrated that prevalence of avian malaria and other haemosporidian parasites is estimated equally well by microscopy and currently used nested PCR-based methods. Both methods have advantages and disadvantages. So we encourage using both these tools in parallel during studies of haemosporidians. Lineages for molecular identification of P. relictum (lineage SGS1) and P. circumflexum (TURDUS1) were determined, also new methods of single cell dissection, DNA extraction and PCR-based analysis of avian malaria and closely related blood parasites were... [to full text] / Paukščių maliariniai parazitai sukelia pavojingas naminių ir laukinių paukščių ligas. Šie parazitai dažnai sutinkami įvairiose paukščių grupėse ir yra plačiai paplitę Europoje, tame tarpe ir Baltijos regione. Dabartinių maliarinių parazitų studijų ypatumas yra tas, kad informacija apie šių parazitų ekologiją, paplitimą, įvairovę ir kitus biologijos aspektus yra surenkama iš laisvai gyvenančių paukščių. Vertinant maliarinių parazitų įtaką paukščių fizinei būklei, elgsenai, lytinei atrankai ir parazitų-šeimininkų ko-evoliucijai yra būtina surinkti informaciją apie maliarinių parazitų specifiškumą, virulentiškumą ir parazitemijos vystymąsi skirtinguose paukščiuose. Tokie duomenys gali būti gaunami atliekant eksperimentinius tyrimus. Deja, tokių studijų kol kas yra nedaug. Disertacijos tikslas – lauko ir eksperimentinių tyrimų pagalba surinkti naują medžiagą apie paukščių maliarinių parazitų biologiją bei susieti PGR paremtais metodais gautą informaciją su tradicinės parazitologijos duomenimis. Šių tyrimų metu įrodyta, kad PGR paremtų ir tradicinės parazitologijos (mikroskopijos) metodų, naudojamų tiriant paukščių maliarinius parazitus ir kitas hemosporidijas, tikslumas nesiskiria. Abu metodai turi savų privalumų ir trūkumų, todėl rekomenduojame naudoti mikroskopijos ir PGR paremtų metodų kombinaciją. Taip pat išvystytas Plasmodium relictum (linija SGS1) ir P. circumflexum (TURDUS1) molekulinis identifikavimas, bei sukurti nauji paukščių maliarinių ir susijusių kraujo parazitų... [toliau žr. visą tekstą]

Severe manifestations of malaria, including cerebral malaria (CM) and respiratory distress, result in approximately three million deaths annually worldwide. Currently, relatively little is known about severe disease pathogenesis. The development and outcome of severe malaria is determined by host-pathogen interactions, a complex interface of genetics and immune responses. Hypothetically, a spectrum of genetic susceptibility and resistance to severe disease exists within the host population, and malaria infection results in diverse host and parasite responses that impact disease outcome. The aim of this study was to identify differential host and parasite responses in a murine model of severe malaria, Plasmodium berghei ANKA (PbA), in CM-susceptible and CM-resistant mice; and to analyze host genetics in patients with severe disease due to Plasmodium falciparum. In vivo, expression microarray analysis showed that, in malaria target organs, differential responses were related to immune response – primarily interferon and complement pathways – and apoptosis. Histopathological examination of the brain confirmed an increased prevalence of apoptosis in CM-susceptible mice. Further examination of the role of complement in CM-susceptibility determined that early complement 5 (C5) activation conferred susceptibility to CM, and that C5 deficiency conferred resistance, which could be recapitulated by antibody blockade of activated C5 or its receptor in susceptible mice. Additionally, single nucleotide polymorphism (SNP) studies identified that complement receptor 1 SNPs were associated with disease severity in patients with P. falciparum malaria. PbA parasites displayed a unique transcriptional signature in each tissue examined (brain, liver, spleen and lung), showed differential gene expression between CM-resistant and susceptible hosts, and were most prominent in lung tissue. Closer examination of lung involvement in PbA infection revealed that PbA-infected C57BL/6 mice develop acute lung injury (ALI), defined by disruption of the alveolar-capillary membrane barrier. ALI susceptibility did not correlate with CM susceptibility, but was influenced by peripheral parasite burden and CD36-mediated parasite sequestration in the lung. PbA provides a clinically relevant experimental model for CM and ALI, through which important disease mechanisms can be identified and modulated. Ideally, the use of such models aids in the discovery of disease biomarkers and novel therapeutic strategies, which may be applied to human severe and cerebral malaria.

Severe manifestations of malaria, including cerebral malaria (CM) and respiratory distress, result in approximately three million deaths annually worldwide. Currently, relatively little is known about severe disease pathogenesis. The development and outcome of severe malaria is determined by host-pathogen interactions, a complex interface of genetics and immune responses. Hypothetically, a spectrum of genetic susceptibility and resistance to severe disease exists within the host population, and malaria infection results in diverse host and parasite responses that impact disease outcome. The aim of this study was to identify differential host and parasite responses in a murine model of severe malaria, Plasmodium berghei ANKA (PbA), in CM-susceptible and CM-resistant mice; and to analyze host genetics in patients with severe disease due to Plasmodium falciparum. In vivo, expression microarray analysis showed that, in malaria target organs, differential responses were related to immune response – primarily interferon and complement pathways – and apoptosis. Histopathological examination of the brain confirmed an increased prevalence of apoptosis in CM-susceptible mice. Further examination of the role of complement in CM-susceptibility determined that early complement 5 (C5) activation conferred susceptibility to CM, and that C5 deficiency conferred resistance, which could be recapitulated by antibody blockade of activated C5 or its receptor in susceptible mice. Additionally, single nucleotide polymorphism (SNP) studies identified that complement receptor 1 SNPs were associated with disease severity in patients with P. falciparum malaria. PbA parasites displayed a unique transcriptional signature in each tissue examined (brain, liver, spleen and lung), showed differential gene expression between CM-resistant and susceptible hosts, and were most prominent in lung tissue. Closer examination of lung involvement in PbA infection revealed that PbA-infected C57BL/6 mice develop acute lung injury (ALI), defined by disruption of the alveolar-capillary membrane barrier. ALI susceptibility did not correlate with CM susceptibility, but was influenced by peripheral parasite burden and CD36-mediated parasite sequestration in the lung. PbA provides a clinically relevant experimental model for CM and ALI, through which important disease mechanisms can be identified and modulated. Ideally, the use of such models aids in the discovery of disease biomarkers and novel therapeutic strategies, which may be applied to human severe and cerebral malaria.

<p>The action of Mung Bean Nuclease (MBN) on DNA makes it possible to clone intact gene fragments from genes of the malaria parasite, Plasmodium. This &ldquo / genease&rdquo / activity has provided a foundation for further investigation of the coding elements of the Plasmodium genome. MBN has been reported to cleave genomic DNA of Plasmodium preferentially at positions before and after genes, but not within gene coding regions. This mechanism has overcome the difficulty encountered in obtaining genes with low expression levels because the cleavage mechanism of the enzyme yields sequences of genes from genomic DNA rather than mRNA. However, as potentially useful as MBN may be, evidence to support its genease activity comes from analysis of a limited number of genes. It is not clear whether this mechanism is specific to certain genes or species of Plasmodia or whether it is a general cleavage mechanism for Plasmodium DNA .There have also been some projects (Nomura et al., 2001 / van Lin, Janse, and Waters, 2000) which have identified MBN generated fragments which contain fragments of genes with both introns and exons, rather than the intact genes expected from MBN-digestion of genomic DNA, which raises concerns about the efficiency of the MBN mechanism in generating complete genes.</p> <p><br /> Using a large-scale, whole genome mapping approach, 7242 MBN generated genome survey sequences (GSSs) have been mapped to determine their position relative to coding sequences within the complete genome sequences of the human malaria parasite Plasmodium falciparum and the incomplete genome of a rodent malaria parasite Plasmodium berghei. The location of MBN cleavage sites was determined with respect to coding regions in orthologous genes, non-coding /intergenic regions and exon-intron boundaries in these two species of Plasmodium. The survey illustrates that for P. falciparum 79% of GSSs had at least one terminal mapping within an ortholog coding sequence and 85% of GSSs which overlapped coding sequence boundaries mapped within 50 bp of the start or end of the gene. Similarly, despite the partial nature of P.berghei genome sequence information, 73% of P.berghei GSSs had at least one terminal mapping within an ortholog coding sequence and 37% of these mapped between 0-50 bp of the start or end of the gene. This indicates that a larger percentage of cleavage sites in both P.falciparum and P.berghei were found proximal to coding regions. Furthermore, 86% of P.falciparum GSSs had at least one terminal mapping within a coding exon and 85% of GSSs which overlapped exon-intron boundaries mapped within 50bp of the exon start and end site. The fact that 11% of GSSs mapped completely to intronic regions, suggests that some introns contain specific cleavage sites sensitive to cleavage and this also indicates that MBN cleavage of Plasmodium DNA does not always yield complete exons.</p> <p><br /> Finally, the results presented herein were obtained from analysis of several thousand Plasmodium genes which have different coding sequences, in different locations on individual chromosomes/contigs in two different species of Plasmodium. Therefore it appears that the MBN mechanism is neither species specific nor is it limited to specific genes.</p>

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 7 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Thesis (Ph.D. (Tropical Medicine))--Mahidol University, 2003.

Thesis (M.C.T.M. (Tropical Pediatrics))--Mahidol University, 2004.

Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (leaves 135-171).

Thesis (Ph. D.)--University of Washington, 1996. / Vita. Includes bibliographical references (leaves [135]-144).