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Now showing 1 - 4 of 4
  • PublicationOpen Access
    An Attenuated HSV-1-Derived Malaria Vaccine Expressing Liver-Stage Exported Proteins Induces Sterilizing Protection against Infectious Sporozoite Challenge
    (2022-02-01T00:00:00Z) Rider, Paul J. F.; KAMIL, MOHD; Yilmaz, Ilknur; Atmaca, Habibe N.; Kalkan-Yazici, Merve; Ziya Doymaz, Mehmet; Kousoulas, Konstantin G.; ALY, Ahmed Sayed Ibrahım; KAMIL, MOHD; YAZICI, MERVE; ALY, AHMED SAYED IBRAHıM
    Here, we present the construction of an attenuated herpes simplex virus type-1 (HSV-1)-vectored vaccine, expressing three liver-stage (LS) malaria parasite exported proteins (EXP1, UIS3 and TMP21) as fusion proteins with the VP26 viral capsid protein. Intramuscular and subcutaneous immunizations of mice with a pooled vaccine, composed of the three attenuated virus strains expressing each LS antigen, induced sterile protection against the intravenous challenge of Plasmodium yoelii 17X-NL salivary gland sporozoites. Our data suggest that this malaria vaccine may be effective in preventing malaria parasite infection using practical routes of immunization in humans.
  • PublicationOpen Access
    Automated wide-field malaria parasite infection detection using Fourier ptychography on stain-free thin-smears
    (2022-07-01T00:00:00Z) AKÇAKIR, OSMAN; Celebi, Lutfi Kadir; KAMIL, MOHD; ALY, Ahmed Sayed Ibrahım; AKÇAKIR, OSMAN; KAMIL, MOHD; ALY, AHMED SAYED IBRAHıM
    Diagnosis of malaria in endemic areas is hampered by the lack of a rapid, stain-free and sensitive method to directly identify parasites in peripheral blood. Herein, we report the use of Fourier ptychography to generate wide-field high-resolution quantitative phase images of erythrocytes infected with malaria parasites, from a whole blood sample. We are able to image thousands of erythrocytes (red blood cells) in a single field of view and make a determination of infection status of the quantitative phase image of each segmented cell based on machine learning (random forest) and deep learning (VGG16) models. Our random forest model makes use of morphology and texture based features of the quantitative phase images. In order to label the quantitative images of the cells as either infected or uninfected before training the models, we make use of a Plasmodium berghei strain expressing GFP (green fluorescent protein) in all life cycle stages. By overlaying the fluorescence image with the quantitative phase image we could identify the infected subpopulation of erythrocytes for labelling purposes. Our machine learning model (random forest) achieved 91% specificity and 72% sensitivity while our deep learning model (VGG16) achieved 98% specificity and 57% sensitivity. These results highlight the potential for quantitative phase imaging coupled with artificial intelligence to develop an easy to use platform for the rapid and sensitive diagnosis of malaria.
  • PublicationMetadata only
    Mitochondrial Spermidine Synthase is Essential for Blood-stage growth of the Malaria Parasite
    (2022-12-01T00:00:00Z) KAMIL, MOHD; KINA, ÜMİT YAŞAR; DEVECİ, GÖZDE; Akyuz, Sevim N.; Yilmaz, Ilknur; ALY, Ahmed Sayed Ibrahım; KAMIL, MOHD; KINA, ÜMİT YAŞAR; DEVECİ, GÖZDE; ALY, AHMED SAYED IBRAHıM
    © 2022Positively-charged polyamines are essential molecules for the replication of eukaryotic cells and are particularly important for the rapid proliferation of parasitic protozoa and cancer cells. Unlike in Trypanosoma brucei, the inhibition of the synthesis of intermediate polyamine Putrescine caused only partial defect in malaria parasite blood-stage growth. In contrast, reducing the intracellular concentrations of Spermidine and Spermine by polyamine analogs caused significant defects in blood-stage growth in Plasmodium yoelii and P. falciparum. However, little is known about the synthesizing enzyme of Spermidine and Spermine in the malaria parasite. Herein, malaria parasite conserved Spermidine Synthase (SpdS) gene was targeted for deletion/complementation analyses by knockout/knock-in constructs in P. yoelii. SpdS was found to be essential for blood-stage growth. Live fluorescence imaging in blood-stages and sporozoites confirmed a specific mitochondrial localization, which is not known for any polyamine-synthesizing enzyme so far. This study identifies SpdS as an excellent drug targeting candidate against the malaria parasite, which is localized to the parasite mitochondrion.
  • PublicationOpen Access
    Highly Sensitive and Rapid Characterization of the Development of Synchronized Blood Stage Malaria Parasites Via Magneto-Optical Hemozoin Quantification
    (2019-10-01T00:00:00Z) Pukancsik, Maria; Molnar, Petra; Orban, Agnes; Butykai, Adam; Marton, Livia; Kezsmarki, Istvan; Vertessy, Beata G.; KAMIL, MOHD; Abraham, Amanah; ALY, AHMED SAYED IBRAHIM; KAMIL, MOHD; ALY, AHMED SAYED IBRAHıM
    The rotating-crystal magneto-optical diagnostic (RMOD) technique was developed as a sensitive and rapid platform for malaria diagnosis. Herein, we report a detailed in vivo assessment of the synchronized Plasmodium vinckei lentum strain blood-stage infections by the RMOD method and comparing the results to the unsynchronized Plasmodium yoelii 17X-NL (non-lethal) infections. Furthermore, we assess the hemozoin production and clearance dynamics in chloroquine-treated compared to untreated self-resolving infections by RMOD. The findings of the study suggest that the RMOD signal is directly proportional to the hemozoin content and closely follows the actual parasitemia level. The lack of long-term accumulation of hemozoin in peripheral blood implies a dynamic equilibrium between the hemozoin production rate of the parasites and the immune system-s clearing mechanism. Using parasites with synchronous blood stage cycle, which resemble human malaria parasite infections with Plasmodium falciparum and Plasmodium vivax, we are demonstrating that the RMOD detects both hemozoin production and clearance rates with high sensitivity and temporal resolution. Thus, RMOD technique offers a quantitative tool to follow the maturation of the malaria parasites even on sub-cycle timescales.