Plasmodium berghei-infected red cells sorted according to DNA content

Parasitology ◽  
1979 ◽  
Vol 78 (3) ◽  
pp. 263-270 ◽  
Author(s):  
R. J. Howard ◽  
F. L. Battye
Keyword(s):  
Red Blood Cells ◽  
Dna Content ◽  
Cell Sorting ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Dye Uptake ◽  
Direct Proportion ◽  
Mouse Blood ◽  
Infected Blood ◽  
A Cell

SUMMARYA cell-sorting method is described for the analysis and separation of red blood cells in Plasmodium berghei-infected mouse blood based on their DNA content. This method involves a selective uptake of the bis-benzimidazole dye 33258 Hoechst, a DNA-binding dye, by red blood cells containing parasites. Infected blood is incubated at 37 °C with the dye then washed at 4 °C to remove unbound dye. Uninfected cells are then non-fluorescent at the characteristic wavelengths for 33258 Hoechst excitation and emission, whereas parasitized cells display fluorescence intensities in approximately direct proportion to the number of parasite nuclei (i.e. amount of parasite DNA) within the cell and can be sorted accordingly. Providing cells were incubated in a complex nutrient medium during dye uptake at 37°C, the sorted parasite cells produced lethal P. berghei infections when injected into BALB/c mice. The dyelabelling technique is simple and sufficient red blood cells at various stages of infection can be collected for biochemical or immunochemical studies by cell sorting.

scholarly journals Plasmodium-infected blood cells analyzed and sorted by flow fluorimetry with the deoxyribonucleic acid binding dye 33258 Hoechst.

1979 ◽  
Vol 27 (4) ◽  
pp. 803-813 ◽  
Author(s):  
R J Howard ◽  
F L Battye ◽  
G F Mitchell
Keyword(s):  
Deoxyribonucleic Acid ◽  
Cell Distribution ◽  
Optimal Conditions ◽  
Dye Uptake ◽  
Direct Proportion ◽  
Mouse Blood ◽  
Infected Blood ◽  
Infected Cells ◽  
Fluorescent Cells

Red cells from Plasmodium berghei infected mouse blood can be sorted on the basis of their DNA content with the bisbenzimidazole dye 33258 Hoechst. The optimal conditions for dye uptake have been established and with these conditions uninfected cells are nonfluorescent and can be completely separated from infected cells which exhibit fluorescence in almost direct proportion to the number of parasite nuclei (i.e. DNA) they contain. The number of fluorescent cells detected and their fluorescence intensity is shown to be dependent on the dye concentration and the incubation medium being used. At least a proportion of the infected cells sorted from each fluorescence peak in the cell distribution retain their infectivity in vivo with some, but not all, conditions of labeling. This technique is being used to separate minor cell populations from infected blood for biochemical and immunochemical analyses and to screen human samples for malaria infected cells.

Changes in the membrane microviscosity of mouse red blood cells infected with Plasmodium berghei detected using n-(9-anthroyloxy) fatty acid fluorescent probes

Parasitology ◽  
1980 ◽  
Vol 80 (2) ◽  
pp. 331-342 ◽  
Author(s):  
R. J. Howard ◽  
W. H. Sawyer
Keyword(s):  
Fatty Acids ◽  
Red Blood Cells ◽  
Outer Membrane ◽  
Fluorescent Probes ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Normal Mouse ◽  
Membrane Microviscosity ◽  
Infected Blood ◽  
Infected Cells

SummaryA set of n-(9-anthroyloxy) fatty acids (n = 2, 6, 9, 12, 16) have been used as fluorescent probes to examine the lipid environment at different depths in the outer membrane of normal mouse erythrocytes and red blood cells from Plasmodium berghei-infected blood. Fluorescent polarization experiments with normal mouse erythrocytes have demonstrated a typical gradient in microviscosity from the surface to the centre of the bilayer as a consequence of the motional properties of the C-atoms of the phospholipid acyl chains. The fluorescent probes rotate faster in the membrane of purified pluriparasitized cells (> 90% purity) than with the remaining fraction of red blood cells from infected blood (20–40% immature, infected red cells, and uninfected red cells), or normal mouse erythrocytes. This increase in fluidity with heavily infected cells occurs predominantly at the centre of the lipid bilayer, rather than at the membrane surface. A comparison of the polarization values of intact and lysed infected cells indicates that the fluorescent fatty acids preferentially label the plasma membrane rather than the internal membranes of infected cells. The results suggest that P. berghei infection causes a change in the composition and/or organization of the outer membrane of pluriparasitized cells which produces a decrease in membrane microviscosity.

Abstract 3680: Transfusion-Induced Tissue Hypoxia is Ameliorated by S -nitrosohemoglobin Repletion of Stored Red Blood Cells

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Diana L Diesen ◽  
Jonathan S Stamler
Keyword(s):  
Animal Models ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Tissue Oxygenation ◽  
Ex Vivo ◽  
Tissue Hypoxia ◽  
Mouse Blood ◽  
Mortality And Morbidity ◽  
Stored Blood ◽  
Related Mortality

Transfusion of stored red blood cells (RBCs) is associated with a decrease in tissue oxygenation in animal models and with increased mortality and morbidity in patients. Recent studies have demonstrated that stored RBCs are deficient in vasodilatory ability and depleted of S -nitrosohemoglobin (SNO-Hb), and that renitrosylation ex vivo can increase SNO-Hb levels and restore vasoactivity. We have examined in a mouse model the extent to which transfusion impairs tissue oxygenation and whether SNO-Hb repletion can ameliorate that impairment. We report here that transfusion of (mouse) RBCs stored for 1 day or 1 week results in tissue hypoxia that is largely prevented by SNO-Hb repletion prior to transfusion ( 1 day stored blood : % decrease in oxygenation 58+/−10% untreated vs. 92+/−0.7% SNO-Hb repleted, p<0.05, n=3– 6; 1 week stored blood : % decrease in oxygenation 66+/−10% untreated vs. 91+/−2.8% SNO-Hb repleted, p<0.05, n=3– 6). Storage of mouse blood beyond human expiration-equivalents (1 month) resulted in substantial lysis and the death of all mice transfused (native and SNO-Hb repleted blood, n=5). In conclusion, repletion of SNO-Hb ameliorates the decrease in tissue oxygenation that results from transfusion of untreated stored blood. Therefore, SNO-Hb repletion may provide a simple and efficacious method to reduce transfusion-related mortality and morbidity.

scholarly journals Comparative assessment of Chemosuppressive activity of three Chinese teas on Plasmodium berghei infected mice

2018 ◽  
Vol 7 (4) ◽  
pp. 376-383
Author(s):  
Bankole Olukayode Olusola ◽  
◽  
Oderinde Abdulganiyu Olumuyiwa ◽  
Keyword(s):  
Body Weight ◽  
Red Blood Cells ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Haemoglobin Concentration ◽  
Treated Group ◽  
Antimalarial Drugs ◽  
Comparative Assessment ◽  
Haematological Parameters ◽  
Weight Changes

Malaria, a hazardous infirmity caused by a parasitic malady of the red blood cells, is without question harming to the wellbeing. In the present investigation, the chemosuppresive and haematopoietic activities of 200 mg/kg and 400 mg/kg body weight of unrefined ethanolic concentrates of three Chinese green teas (BIA 849, TD 570 and GB/T19598) were assessed using the 4-day suppressive anti-plasmodial assay in mice Plasmodium berghei (NK65 strain) pre-infected mice. The effect of the extracts on weight of the animals was evaluated. It was observed that 200 mg/kg bw (body weight) of BIA 849 and GB/T19598 were as potent as 5 mg/kg bw of chloroquine, with percentage suppressions of 58.97 ± 5.04, 57.63 ± 5.62 and 57.50 ± 4.5, respectively. TD570 at 200 mg/kg bw was more effective in suppressing plasmodium. 400 mg/kg body weight of TD570 and GB/T19598 extracts were more potent than 5 mg/kg bw of chloroquine having 100 % chemosuppression. The chemosuppression of BIA 849 did not change altogether at 400 mg/kg bw. The haematological parameters, WBC, RBC and MCV did not significantly change in the groups treated with the tea extracts utilizing suppressive model of malaria treatment contrasted with the uninfected group and were comparable to those treated with chloroquine. Haemoglobin concentration nonetheless, varied significantly with respect to the uninfected group. Weight changes were most significant with 200 mg/kg bw of TD 570 treated group (32 % increase) on suppression. All in all, the green teas displayed high chemosuppressive and haematopoietic possibilities and are thusly prescribed as contender for additionally screening as elective antimalarial drugs

scholarly journals Merocyanine 540-sensitized photoinactivation of human erythrocytes parasitized by Plasmodium falciparum

Blood ◽  
1992 ◽  
Vol 80 (1) ◽  
pp. 21-24 ◽  
Author(s):  
OM Smith ◽  
SA Dolan ◽  
JA Dvorak ◽  
TE Wellems ◽  
F Sieber
Keyword(s):  
Stem Cells ◽  
Plasmodium Falciparum ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Human Blood ◽  
Blood Cells ◽  
Potential Usefulness ◽  
Hematopoietic Stem ◽  
Infected Blood ◽  
Merocyanine 540

The purpose of this study was to evaluate the photosensitizing dye merocyanine 540 (MC540) as a means for extracorporeal purging of Plasmodium falciparum-infected erythrocytes from human blood. Parasitized red blood cells bound more dye than nonparasitized cells, and exposure to MC540 and light under conditions that are relatively well tolerated by normal erythrocytes and normal pluripotent hematopoietic stem cells reduced the concentration of parasitized cells by as much as 1,000-fold. Cells parasitized by the chloroquine- sensitive HB3 clone and the chloroquine-resistant Dd2 clone of P falciparum were equally susceptible to MC540-sensitized photolysis. These data suggest the potential usefulness of MC540 in the purging of P falciparum-infected blood.

scholarly journals Deformability based sorting of stored red blood cells reveals donor-dependent aging curves

Lab on a Chip ◽  
2020 ◽  
Vol 20 (2) ◽  
pp. 226-235 ◽  
Author(s):  
Emel Islamzada ◽  
Kerryn Matthews ◽  
Quan Guo ◽  
Aline T. Santoso ◽  
Simon P. Duffy ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Cold Storage ◽  
Red Blood Cell ◽  
Cell Sorting ◽  
Blood Cells ◽  
Cell Deformability ◽  
Red Blood Cell Deformability

Cell sorting using microfluidic ratchets enables sensitive and consistent characterization of donor red blood cell deformability. Using this capability, we show the degradation of red blood cell deformability during cold storage is donor-dependent.

Biomechanical properties of red blood cells infected by Plasmodium berghei ANKA

2019 ◽  
Vol 234 (11) ◽  
pp. 20546-20553 ◽  
Author(s):  
Sangwoo Kwon ◽  
Dong‐Hun Lee ◽  
Se‐Jik Han ◽  
Woochul Yang ◽  
Fu‐Shi Quan ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Biomechanical Properties ◽  
Plasmodium Berghei Anka

Superoxide Dismutase (SOD) in Mouse Red Blood Cells Infected with Plasmodium berghei

1982 ◽  
Vol 68 (2) ◽  
pp. 337 ◽  
Author(s):  
Upsorn Suthipark ◽  
Jerapan Krungkrai ◽  
Amornrat Jearnpipatkul ◽  
Yongyuth Yuthavong ◽  
Bhinyo Panijpan
Keyword(s):  
Superoxide Dismutase ◽  
Red Blood Cells ◽  
Plasmodium Berghei ◽  
Blood Cells

Establishment of a murine model of cerebral malaria in KunMing mice infected with Plasmodium berghei ANKA

Parasitology ◽  
2016 ◽  
Vol 143 (12) ◽  
pp. 1672-1680 ◽  
Author(s):  
YAN DING ◽  
WENYUE XU ◽  
TAOLI ZHOU ◽  
TAIPING LIU ◽  
HONG ZHENG ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Cerebral Malaria ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Mononuclear Cells ◽  
Clinical Signs ◽  
Necrosis Factor Alpha ◽  
Experimental Cerebral Malaria ◽  
Km Mice

SUMMARYMalaria remains one of the most devastating diseases. Cerebral malaria (CM) is a severe complication of Plasmodium falciparum infection resulting in high mortality and morbidity worldwide. Analysis of precise mechanisms of CM in humans is difficult for ethical reasons and animal models of CM have been employed to study malaria pathogenesis. Here, we describe a new experimental cerebral malaria (ECM) model with Plasmodium berghei ANKA infection in KunMing (KM) mice. KM mice developed ECM after blood-stage or sporozoites infection, and the development of ECM in KM mice has a dose-dependent relationship with sporozoites inoculums. Histopathological findings revealed important features associated with ECM, including accumulation of mononuclear cells and red blood cells in brain microvascular, and brain parenchymal haemorrhages. Blood–brain barrier (BBB) examination showed that BBB disruption was present in infected KM mice when displaying clinical signs of CM. In vivo bioluminescent imaging experiment indicated that parasitized red blood cells accumulated in most vital organs including heart, lung, spleen, kidney, liver and brain. The levels of inflammatory cytokines interferon-gamma, tumour necrosis factor-alpha, interleukin (IL)-17, IL-12, IL-6 and IL-10 were all remarkably increased in KM mice infected with P. berghei ANKA. This study indicates that P. berghei ANKA infection in KM mice can be used as ECM model to extend further research on genetic, pharmacological and vaccine studies of CM.

scholarly journals Sorting same-size red blood cells in deep deterministic lateral displacement devices

2018 ◽  
Vol 859 ◽  
pp. 433-475 ◽  
Author(s):  
Gökberk Kabacaoğlu ◽  
George Biros
Keyword(s):  
Mechanical Properties ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Lateral Displacement ◽  
Flow Direction ◽  
Lateral Direction ◽  
Deterministic Lateral Displacement ◽  
Deformable Particles ◽  
Dense Suspensions ◽  
A Cell

Microfluidic sorting of deformable particles finds many applications, for example, medical devices for cells. Deterministic lateral displacement (DLD) is one of them. Particle sorting via DLD relies only on hydrodynamic forces. For rigid spherical particles, this separation is to a great extent understood and can be attributed to size differences: large particles displace in the lateral direction with respect to the flow while small particles travel in the flow direction with negligible lateral displacement. However, the separation of non-spherical deformable particles such as red blood cells (RBCs) is more complicated than that of rigid particles. For example, is it possible to separate deformable particles that have the same size but different mechanical properties? We study deformability-based sorting of same-size RBCs via DLD using an in-house integral equation solver for vesicle flows in two dimensions. Our goal is to quantitatively characterize the physical mechanisms that enable the cell separation. To this end, we systematically investigate the effects of the interior fluid viscosity and membrane elasticity of a cell on its behaviour. In particular, we consider deep devices in which a cell can show rich dynamics such as taking a particular angular orientation depending on its mechanical properties. We have found out that cells moving with a sufficiently high positive inclination angle with respect to the flow direction displace laterally while those with smaller angles travel with the flow streamlines. Thereby, deformability-based cell sorting is possible. The underlying mechanism here is cell migration due to the cell’s positive inclination and the shear gradient. The higher the inclination is, the farther the cell can travel laterally. We also assess the efficiency of the technique for dense suspensions. It turns out that most of the cells in dense suspensions do not displace in the lateral direction no matter what their deformability is. As a result, separating cells using a DLD device becomes harder.

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