Transcriptional Analysis of Tightly Synchronized Plasmodium falciparum Intraerythrocytic Stages by RT-qPCR

Author(s):  
Núria Casas-Vila ◽  
Anastasia K. Pickford ◽  
Harvie P. Portugaliza ◽  
Elisabet Tintó-Font ◽  
Alfred Cortés
2000 ◽  
Vol 8 (8) ◽  
pp. 350-351 ◽  
Author(s):  
Alister Craig ◽  
Artur Scherf

2010 ◽  
Vol 76 (12) ◽  
pp. 3959-3966 ◽  
Author(s):  
Yu Du ◽  
Jolyn E. Gisselberg ◽  
Jacob D. Johnson ◽  
Patricia J. Lee ◽  
Sean T. Prigge ◽  
...  

ABSTRACT Plasmodium falciparum, in addition to scavenging essential fatty acids from its intra- and intercellular environments, possesses a functional complement of type II fatty acid synthase (FAS) enzymes targeted to the apicoplast organelle. Recent evidence suggests that products of the plasmodial FAS II system may be critical for the parasite's liver-to-blood cycle transition, and it has been speculated that endogenously generated fatty acids may be precursors for essential cofactors, such as lipoate, in the apicoplast. β-Ketoacyl-acyl carrier protein (ACP) synthase III (pfKASIII or FabH) is one of the key enzymes in the initiating steps of the FAS II pathway, possessing two functions in P. falciparum: the decarboxylative thio-Claisen condensation of malonyl-ACP and various acyl coenzymes A (acyl-CoAs; KAS activity) and the acetyl-CoA:ACP transacylase reaction (ACAT). Here, we report the generation and characterization of a hybrid Lactococcus lactis strain that translates pfKASIII instead of L. lactis f abH to initiate fatty acid biosynthesis. The L. lactis expression vector pMG36e was modified for the efficient overexpression of the plasmodial gene in L. lactis. Transcriptional analysis indicated high-efficiency overexpression, and biochemical KAS and ACAT assays confirm these activities in cell extracts. Phenotypically, the L. lactis strain expressing pfKASIII has a growth rate and fatty acid profiles that are comparable to those of the strain complemented with its endogenous gene, suggesting that pfKASIII can use L. lactis ACP as substrate and perform near-normal function in L. lactis cells. This strain may have potential application as a bacterial model for pfKASIII inhibitor prescreening.


2011 ◽  
Vol 12 (6) ◽  
pp. R56 ◽  
Author(s):  
Kate M Broadbent ◽  
Daniel Park ◽  
Ashley R Wolf ◽  
Daria Van Tyne ◽  
Jennifer S Sims ◽  
...  

Author(s):  
D.J.P. Ferguson ◽  
A.R. Berendt ◽  
J. Tansey ◽  
K. Marsh ◽  
C.I. Newbold

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).


1981 ◽  
Vol 46 (02) ◽  
pp. 547-549 ◽  
Author(s):  
E M Essien ◽  
M I Ebhota

SummaryDuring acute malaria infection, platelets in human platelet-rich plasma are hypersensitive to the addition of ADP between 1.0 uM and 5.0 uM, or adrenaline 0.11 uM as aggregating agents. The mean maximum aggregation amplitude (as % of light transmission) obtained from 8 subjects in response to added ADP (1.0 uM), 39.8 ± 27 (1SD), was significantly greater than the value in 6 controls (5.2±6.7 (1SD); t = 3, 51 P <0.005). A similar pattern of response was obtained with higher ADP concentrations (2.4,4.5 or 5.0 uM) in 22 patients and 20 control subjects (89.9±14.9% vs 77.8±16.5% (1SD) t = 2.45, P <0.02). Addition of 4.5 uM ADP to patient PRP usually evoked only a single aggregation wave (fused primary and secondary waves) while the typical primary and secondary wave pattern was usually obtained from controls.Mean plasma B-thromboglobulin (BTG) concentration in 7 patients (208.3 ± 15.6 ng/ml) was significantly higher than the value in 6 control subjects (59.2±15.7 ng/ml; t = 13.44, P <0.002).


2018 ◽  
Vol 18 (05) ◽  
pp. 332-338
Author(s):  
C. Kleine ◽  
U. Ziegler ◽  
E.-M. Schwienhorst ◽  
A. Stich

ZusammenfassungDer folgende Artikel fokussiert auf Erkrankungen, deren Erreger von Vektoren (Insekten, Spinnentieren) aktiv auf den Menschen übertragen werden. Sie spielen in tropischen und subtropischen Regionen der Erde eine erhebliche Rolle und sind auch im Rahmen der Differenzialdiagnose bei kranken Reiserückkehrern von großer Bedeutung. Am wichtigsten ist die Malaria, insbesondere die lebensbedrohliche Malaria tropica durch Plasmodium falciparum. Jede fieberhafte Erkrankung aus den Tropen erfordert eine zeitnahe Malaria-Diagnostik. Tropische Viruserkrankungen durch Dengue-, West-Nil-, Chikungunya- oder Zika-Viren haben sich in den vergangenen Jahrzehnten massiv ausgebreitet und stellen auch eine zunehmende Bedrohung für den europäischen Raum dar. Andere Vektor-übertragene Erkrankungen sind zwar von großer lokaler Relevanz in endemischen Regionen, aber als importierte Infektionen in Deutschland relativ selten. Bei der Betreuung der Patienten empfiehlt sich eine enge Kooperation mit Tropenmedizinern.


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