66 research outputs found
Changes in lipid composition during sexual development of the malaria parasite Plasmodium falciparum
Abstract
Background
The development of differentiated sexual stages (gametocytes) within human red blood cells is essential for the propagation of the malaria parasite, since only mature gametocytes will survive in the mosquito’s midgut. Hence gametocytogenesis is a pre-requisite for transmission of the disease. Physiological changes involved in sexual differentiation are still enigmatic. In particular the lipid metabolism—despite being central to cellular regulation and development—is not well explored.
Methods
Here the lipid profiles of red blood cells infected with the five different sexual stages of Plasmodium falciparum were analysed by mass spectrometry and compared to those from uninfected and asexual trophozoite infected erythrocytes.
Results
Fundamental differences between erythrocytes infected with the different parasite stages were revealed. In mature gametocytes many lipids that decrease in the trophozoite and early gametocyte infected red blood cells are regained. In particular, regulators of membrane fluidity, cholesterol and sphingomyelin, increased significantly during gametocyte maturation. Neutral lipids (serving mainly as caloriometric reserves) increased from 3Â % of total lipids in uninfected to 27Â % in stage V gametocyte infected red blood cells. The major membrane lipid class (phospholipids) decreased during gametocyte development.
Conclusions
The lipid profiles of infected erythrocytes are characteristic for the particular parasite life cycle and maturity stages of gametocytes. The obtained lipid profiles are crucial in revealing the lipid metabolism of malaria parasites and identifying targets to interfere with this deadly disease.We are grateful to the Australian Red Cross for providing human RBCs and
serum. Support of the Australian Research Council is acknowledged. TWM
is an Australian Research Council Future Fellow (FT110100249)
Seasonal change in the daily timing of behaviour of the common vole, Microtus arvalis
1. Seasonal effects on daily activity patterns in the common vole were established by periodic trapping in the field and continuous year round recording of running wheel and freeding activity in cages exposed to natural meteorological conditions.
2. Trapping revealed decreased nocturnality in winter as compared to summer. This was paralelled by a winter reduction in both nocturnal wheel running and feeding time in cages.
3. Frequent trap checks revealed a 2 h rhythm in daytime catches in winter, not in summer. Cage feeding activity in daytime was always organized in c. 2 h intervals, but day-to-day variations in phase blurred the rhythm in summer in a summation of individual daily records. Thus both seasonal and short-term temporal patterns are consistent between field trappings and cage feeding records.
4. Variables associated with the seasonal change in daily pattern were: reproductive state (sexually active voles more nocturnal), age (juveniles more nocturnal), temperature (cold days: less nocturnal), food (indicated by feeding experiments), habitat structure (more nocturnal in habitat with underground tunnels).
5. Minor discrepancies between field trappings and cage feeding activity can be explained by assuming increased trappability of voles in winter. Cage wheel running is not predictive of field trapping patterns and is thought to reflect behavioral motivations not associated with feeding but with other activities (e.g., exploratory, escape, interactive behaviour) undetected by current methods, including radiotelemetry and passage-counting.
6. Winter decrease in nocturnality appears to involve a reduction in nocturnal non-feeding and feeding behaviour and is interpreted primarily as an adaptation to reduce energy expenditure in adverse but socially stable winter conditions.
The Plasmodium Export Element Revisited
We performed a bioinformatical analysis of protein export elements (PEXEL) in the putative proteome of the malaria parasite Plasmodium falciparum. A protein family-specific conservation of physicochemical residue profiles was found for PEXEL-flanking sequence regions. We demonstrate that the family members can be clustered based on the flanking regions only and display characteristic hydrophobicity patterns. This raises the possibility that the flanking regions may contain additional information for a family-specific role of PEXEL. We further show that signal peptide cleavage results in a positional alignment of PEXEL from both proteins with, and without, a signal peptide
clag9 Is Not Essential for PfEMP1 Surface Expression in Non-Cytoadherent Plasmodium falciparum Parasites with a Chromosome 9 Deletion
BACKGROUND: The expression of the clonally variant virulence factor PfEMP1 mediates the sequestration of Plasmodium falciparum infected erythrocytes in the host vasculature and contributes to chronic infection. Non-cytoadherent parasites with a chromosome 9 deletion lack clag9, a gene linked to cytoadhesion in previous studies. Here we present new clag9 data that challenge this view and show that surface the non-cytoadherence phenotype is linked to the expression of a non-functional PfEMP1. METHODOLOGY/PRINCIPAL FINDINGS: Loss of adhesion in P. falciparum D10, a parasite line with a large chromosome 9 deletion, was investigated. Surface iodination analysis of non-cytoadherent D10 parasites and COS-7 surface expression of the CD36-binding PfEMP1 CIDR1α domain were performed and showed that these parasites express an unusual trypsin-resistant, non-functional PfEMP1 at the erythrocyte surface. However, the CIDR1α domain of this var gene expressed in COS-7 cells showed strong binding to CD36. Atomic Force Microscopy showed a slightly modified D10 knob morphology compared to adherent parasites. Trafficking of PfEMP1 and KAHRP remained functional in D10. We link the non-cytoadherence phenotype to a chromosome 9 breakage and healing event resulting in the loss of 25 subtelomeric genes including clag9. In contrast to previous studies, knockout of the clag9 gene from 3D7 did not interfere with parasite adhesion to CD36. CONCLUSIONS/SIGNIFICANCE: Our data show the surface expression of non-functional PfEMP1 in D10 strongly indicating that genes other than clag9 deleted from chromosome 9 are involved in this virulence process possibly via post-translational modifications
Investigating the Host Binding Signature on the Plasmodium falciparum PfEMP1 Protein Family
The Plasmodium falciparum erythrocyte membrane protein 1
(PfEMP1) family plays a central role in antigenic variation and cytoadhesion of
P. falciparum infected erythrocytes. PfEMP1
proteins/var genes are classified into three main
subfamilies (UpsA, UpsB, and UpsC) that are hypothesized to have different roles
in binding and disease. To investigate whether these subfamilies have diverged
in binding specificity and test if binding could be predicted by adhesion domain
classification, we generated a panel of 19 parasite lines that primarily
expressed a single dominant var transcript and assayed binding
against 12 known host receptors. By limited dilution cloning, only UpsB and UpsC
var genes were isolated, indicating that UpsA
var gene expression is rare under in vitro
culture conditions. Consequently, three UpsA variants were obtained by rosette
purification and selection with specific monoclonal antibodies to create a more
representative panel. Binding assays showed that CD36 was the most common
adhesion partner of the parasite panel, followed by ICAM-1 and TSP-1, and that
CD36 and ICAM-1 binding variants were highly predicted by adhesion domain
sequence classification. Binding to other host receptors, including CSA, VCAM-1,
HABP1, CD31/PECAM, E-selectin, Endoglin, CHO receptor “X”, and
Fractalkine, was rare or absent. Our findings identify a category of larger
PfEMP1 proteins that are under dual selection for ICAM-1 and CD36 binding. They
also support that the UpsA group, in contrast to UpsB and UpsC
var genes, has diverged from binding to the major
microvasculature receptor CD36 and likely uses other mechanisms to sequester in
the microvasculature. These results demonstrate that CD36 and ICAM-1 have left
strong signatures of selection on the PfEMP1 family that can be detected by
adhesion domain sequence classification and have implications for how this
family of proteins is specializing to exploit hosts with varying levels of
anti-malaria immunity
Changes in lipid composition during sexual development of the malaria parasite Plasmodium falciparum
Human malarial disease: a consequence of inflammatory cytokine release
Malaria causes an acute systemic human disease that bears many similarities, both clinically and mechanistically, to those caused by bacteria, rickettsia, and viruses. Over the past few decades, a literature has emerged that argues for most of the pathology seen in all of these infectious diseases being explained by activation of the inflammatory system, with the balance between the pro and anti-inflammatory cytokines being tipped towards the onset of systemic inflammation. Although not often expressed in energy terms, there is, when reduced to biochemical essentials, wide agreement that infection with falciparum malaria is often fatal because mitochondria are unable to generate enough ATP to maintain normal cellular function. Most, however, would contend that this largely occurs because sequestered parasitized red cells prevent sufficient oxygen getting to where it is needed. This review considers the evidence that an equally or more important way ATP deficency arises in malaria, as well as these other infectious diseases, is an inability of mitochondria, through the effects of inflammatory cytokines on their function, to utilise available oxygen. This activity of these cytokines, plus their capacity to control the pathways through which oxygen supply to mitochondria are restricted (particularly through directing sequestration and driving anaemia), combine to make falciparum malaria primarily an inflammatory cytokine-driven disease
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