In vitro Cultivation of Blood Cells of Drosophila melanogaster in a Synthetic Medium

1. Methods of recovering adequate amounts of Plasmodium knowlesi from the monkey (Macaca, mulatta) for biochemical studies and in vitro cultivation are described. Concentrates of red blood cells parasitized with P. knowlesi can be obtained by differential sedimentation of parasitized blood because of physical and chemical changes produced by the parasites in the host cell and the plasma of the blood. 2. Two different techniques, the rocker-dilution and the rocker-perfusion methods, are described for the cultivation of malarial parasites. Details of the apparatus, assembly, and sterilization are given, as well as methods of counting and evaluating parasites. 3. In a series of 235 control experiments for 20 to 24 hours using three types of apparatus, the average rate of multiplication was 3.9. Each technique has specific value for studying the various aspects of metabolism, nutrition, and the action of antimalarial drugs.

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MAINTENANCE OF IMAGINAL DISCS OF DROSOPHILA MELANOGASTER IN CHEMICALLY DEFINED MEDIA

The Journal of Cell Biology ◽

10.1083/jcb.41.3.876 ◽

1969 ◽

Vol 41 (3) ◽

pp. 876-885 ◽

Cited By ~ 215

Author(s):

James A. Robb

Keyword(s):

Protein Synthesis ◽

Drosophila Melanogaster ◽

Synthetic Medium ◽

Sodium Concentration ◽

Imaginal Discs ◽

Magnesium Concentration ◽

Insoluble Protein ◽

Total Rna ◽

Phosphate Buffered Saline

A phosphate-buffered saline and a chemically defined synthetic medium for in vitro maintenance of imaginal discs of Drosophila melanogaster were developed. The composition of the chemically defined medium was varied in order to optimize the incorporation of tritiated uridine into RNA and tritiated amino acids into acid-insoluble protein. The optimal ranges obtained were: pH, 6.75–7.35; osmolarity, 285–345 milliosmoles/liter; sodium concentration, 40–60 mM/liter; potassium concentration, 40–60 mM/liter; magnesium concentration, 0.5–3.5 mM/liter; calcium concentration, 0.3–1.5 mM/liter; and inorganic phosphate concentration, 1.5–4.0 mM/liter. The phosphate-buffered saline is superior to a commonly used insect Ringer solution in maintaining total RNA and acid-insoluble protein synthesis in culture. The chemically defined synthetic medium permits linear total RNA and acid-insoluble protein synthesis for more than 48 hr, DNA synthesis for several hours, normal differentiation to occur after 74 hr in vitro, and trypsinization of imaginal discs into single cell suspensions without developmental damage.

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Study of some parameters affecting the in vitro cultivation of Plasmodium falciparum within saimiri sciureus red blood cells

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761986000200005 ◽

1986 ◽

Vol 81 (2) ◽

pp. 165-170 ◽

Cited By ~ 1

Author(s):

T. Fandeur ◽

J. P. Dedet

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Osmotic Fragility ◽

Culture Conditions ◽

Saimiri Sciureus ◽

In Vitro Cultivation ◽

In Vitro Growth ◽

Serum Supplement

The in vitro growth and multiplication of the erythrocytic stages of Plasmodium falciparum within Saimiri sciureus (squirrel monkey) red blood cells have been studied. Various parameters, such as the origin of the red blood cells and serum supplement, nature of the buffer, influence of the final pH of the medium, role of proteose peptone and glucose addition, were investigated. The selection of the best culture conditions led to the obtention of a reproducible in vitro growth of two parasite cycles in Saimiri erythrocytes, which is an useful achievement for in vitro studies. Our failure to establish a continuous culture line for longer than 19 days, could be explained by a dramatic increasing of osmotic fragility of the Saimiri red blood cells related to their small size.

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Antiplasmodial Activities of Some Locally Prepared Herbs Against Plasmodium falciparum Obtained from Asymptomatic School Children in Zaria, Kaduna State, Nigeria

Nigerian Journal of Pure and Applied Sciences ◽

10.48198/njpas/21.a18 ◽

2021 ◽

pp. 4101-4113

Author(s):

Balogun Islamiat D ◽

Inabo Helen I ◽

Ella Elijah E

Keyword(s):

Plasmodium Falciparum ◽

Blood Cells ◽

Cardiac Glycosides ◽

In Vitro Cultivation ◽

Plasmodium Infection ◽

Statistical Association ◽

Average Percentage ◽

Gender And Age ◽

Phytochemical Constituents

The efficacy of current or any intended antimalarial can only be resolute by cultivation and susceptibility studies. The aim of this research was to cultivate Plasmodium falciparum in vitro and comparing the antiplasmodial effects of standard antimalarial medications including herbaceous preparation. Asymptomatic pupils attending some schools in Zaria, Kaduna state, Nigeria were recruited into this research and blood samples were collected from them. Microscopy was done after thin and thick blood films were prepared and stained. The antiplasmodial activities of antimalarial drugs as well as herbal preparation were determined after the successful culturing of red blood cells in the Jatropha curcas medium.The phytochemical constituents of the herbs that made up the concoction were determined. The incidence of asymptomatic Plasmodium infection amongst school kids was 17.5%. There was no statistical association of location, gender and age with the obtained prevalence. Presence of alkaloids was observed in every plant screened while the same was also observed for tannins except in Enantia chloranta.Steroids and phlobatanins were observed in Citrus aurantifolia while there was presence of saponins in all the plant extracts except Cymbopogbon citratus. All the plants except Enantia chloranta contained flavonoids. There was presence of terpenoids in all screened plants except Azadirachta indica and Cymbopogbon citratus while cardiac glycosides were found in every plant except Magnifera indica and Enantia chloranta. Results obtained from the in vitro cultivation of the Plasmodium falciparum with Athemether/lumefanthrine, amodiaquine and herbal concoction showed average percentage parasite inhibition of 80%, 37.8% and 38.6% respectively. This implies that Arthemether/lumefanthrine was capable of inhibiting the growth of the parasite best. The herbal concoction also inhibited growth (38.6% inhibition). There is need for additional investigation on a wider variety of plants to explore their antiplasmodial activities since there is evidence that it works, and it is quite available and affordable.

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Functional Significance of the Crystal Cells in the Larva of Drosophila melanogaster

The Journal of Cell Biology ◽

10.1083/jcb.5.2.235 ◽

1959 ◽

Vol 5 (2) ◽

pp. 235-240 ◽

Cited By ~ 87

Author(s):

M. T. M. Rizki ◽

Rose M. Rizki

Keyword(s):

Drosophila Melanogaster ◽

Blood Cells ◽

Structural Integrity ◽

Present Report ◽

Enzymatic Reaction ◽

Humoral Factors ◽

To Come ◽

Crystal Cells

Cytoplasmic crystalline inclusions are found in some larval haemocytes of Drosophila melanogaster. Blackening can be experimentally induced in these cells, and previously it was suggested that either the substrate or enzyme for the tyrosine-tyrosinase system leading to melanin production in Drosophila larvae is found in these inclusions in the crystal cells. The present report is an attempt to further localize the enzyme and substrate. Larvae have been fed on food containing α-C14-tyrosine and autoradiographs of the blood cells taken from these larvae subsequently prepared. The C14 activity in the crystal cells is restricted to the crystal inclusions in the cells and is significantly higher than that found in the other type of haemocytes, the plasmatocytes. When samples of the blood cells are incubated in DOPA solution, the extra-crystalline cytoplasm becomes blackened while the crystals themselves remain colorless. These observations are consistent with the hypothesis that the substrate is localized in the crystal inclusions whereas enzyme is found in the surrounding cytoplasm. An in vivo structural isolation would serve to separate enzyme and substrate rather than an inhibition by dehydrogenases as postulated by previous authors. In vitro examination with the phase microscope has shown that the crystal cells rupture easily and the crystals dissolve in the haemolymph. Therefore any treatment which tends to disrupt the structural integrity of the cell will allow the enzyme and substrate to come together. Humoral factors preceding metamorphosis might account for the in vivo release of the enzymatic reaction by initially altering the permeability of the cell.

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Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162132 ◽

1990 ◽

Vol 48 (3) ◽

pp. 914-915

Author(s):

D.J.P. Ferguson ◽

A.R. Berendt ◽

J. Tansey ◽

K. Marsh ◽

C.I. Newbold

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Umbilical Vein ◽

Cell Types ◽

Amelanotic Melanoma ◽

Cytoplasmic Process ◽

Umbilical Vein Endothelial Cells

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).

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Mechanism of the Antiplatelet Action of Dipyridamole in Whole Blood: Modulation of Adenosine Concentration and Activity

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661437 ◽

1986 ◽

Vol 55 (01) ◽

pp. 012-018 ◽

Cited By ~ 86

Author(s):

Paolo Gresele ◽

Jef Arnout ◽

Hans Deckmyn ◽

Jos Vermylen

Keyword(s):

Platelet Aggregation ◽

Adenosine Receptor ◽

Whole Blood ◽

Blood Cells ◽

Inhibitory Action ◽

Phosphodiesterase Inhibitor ◽

Inhibitory Effect ◽

Adenosine Concentration ◽

Pharmacological Studies

SummaryDipyridamole inhibits platelet aggregation in whole blood at lower concentrations than in plasma. The blood cells responsible for increased effectiveness in blood are the erythrocytes. Using the impedance aggregometer we have carried out a series of pharmacological studies in vitro to elucidate the mechanism of action of dipyridamole in whole blood. Adenosine deaminase, an enzyme breaking down adenosine, reverses the inhibitory action of dipyridamole. Two different adenosine receptor antagonists, 5’-deoxy-5’-methylthioadenosine and theophylline, also partially neutralize the activity of dipyridamole in blood. Enprofylline, a phosphodiesterase inhibitor with almost no adenosine receptor antagonistic properties, potentiates the inhibition of platelet aggregation by dipyridamole. An inhibitory effect similar to that of dipyridamole can be obtained combining a pure adenosine uptake inhibitor (RE 102 BS) with a pure phosphodiesterase inhibitor (MX-MB 82 or enprofylline). Mixing the blood during preincubation with dipyridamole increases the degree of inhibition. Lowering the haematocrit slightly reduces the effectiveness.Although we did not carry out direct measurements of adenosine levels, the results of our pharmacological studies clearly show that dipyridamole inhibits platelet aggregation in whole blood by blocking the reuptake of adenosine formed from precursors released by red blood cells following microtrauma. Its slight phosphodiesterase inhibitory action potentiates the effects of adenosine on platelets.

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