THE IN VITRO PREPARATION AND HISTOCHEMICAL PROPERTIES OF SUBSTANCES RESEMBLING CEROID

Substances possessing the same histochemical properties as the ceroid in cirrhotic livers of rats fed choline-deficient diets have been prepared from various unsaturated fats, fatty acids and their esters by autoxidation but could not be obtained from hydrocarbons or saturated fats or fatty acids. The formation of ceroid-like substances occurred first on surfaces or at interfaces in the reaction mixtures. It was inhibited by antioxidants and was accelerated by the addition of tissues, blood cells, erythrocytic stroma, or hemoglobin, by emulsification, by increasing the surface exposed to the air, and by increasing the temperature. Histochemical studies provided much evidence that the following properties of ceroid might be attributed to the products of the autoxidation of unsaturated lipids: insolubility in organic solvents, sudanophilia, yellowing by concentrated nitric acid, positive periodic acid-Schiff's reaction, basophilia, acid fastness, positive hernofuscin reaction, and reduction of diammine silver carbonate and alkaline potassium permanganate. The normal reactivity of cells or tissues embedded in ceroid was effectively masked by the pigment, apparently, initially at least, by preventing the reagents' gaining access to them. It is suggested that the iron sometimes demonstrated in ceroid may be that of blood cells or tissue fragments incompletely masked by the ceroid. It is concluded that whenever conditions are such that unsaturated fats accumulate in tissues to such an extent that a relative lack of biological anti-oxidants results, autoxidation of the fats and their conversion to ceroid pigment are favored, and that ceroid and the lipofuscin pigment of vitamin E deficiency may be fundamentally similar.

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In vitro effects of an azo compound on the haemolysis and unsaturated fatty acids of red blood cells

Clinica Chimica Acta ◽

10.1016/s0009-8981(97)00053-3 ◽

1997 ◽

Vol 263 (2) ◽

pp. 157-164 ◽

Cited By ~ 2

Author(s):

Bilgehan Pekiner ◽

Jack F. Pennock

Keyword(s):

Fatty Acids ◽

Red Blood Cells ◽

Blood Cells ◽

Unsaturated Fatty Acids ◽

Azo Compound ◽

In Vitro Effects

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Effects of in vitro hyperthermia on fatty acids of red blood cells and plasma lipids from patients with multiple sclerosis

Journal of the Neurological Sciences ◽

10.1016/0022-510x(90)90237-h ◽

1990 ◽

Vol 95 (2) ◽

pp. 141-151 ◽

Cited By ~ 2

Author(s):

George D. Cherayil

Keyword(s):

Multiple Sclerosis ◽

Fatty Acids ◽

Red Blood Cells ◽

Blood Cells ◽

Plasma Lipids

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151 LIPID COMPOSITION OF FAT DROPLETS OF IN VIVO- AND IN VITRO PRODUCED PORCINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab151 ◽

2007 ◽

Vol 19 (1) ◽

pp. 193 ◽

Cited By ~ 1

Author(s):

M. Romek ◽

B. Gajda ◽

E. Krzysztofowicz ◽

Z. Smorag

Keyword(s):

Fatty Acids ◽

Free Fatty Acids ◽

Lipid Composition ◽

Early Stage ◽

Nile Blue ◽

Thin Sections ◽

Unsaturated Lipids ◽

Different Types

Early-stage porcine embryos to peri-hatching blastocysts contain high levels of intracellular lipids in the form of fat droplets and are highly sensitive to cryopreservation. Recently, our sterological studies demonstrated that in embryos produced in vivo and cultured in vitro, the volume of lipid droplets significantly decreased from zygote to blastocyst. To date, however, there have been no reports concerning the type of lipids in pig embryos produced in vivo and in vitro. Thus, the objective of the present study was to investigate the lipid composition of fat droplets in pig embryos produced in vivo and in vitro. The experiment was carried out on pig zygotes produced in vivo and 2–4 and 8–16-cell embryos, morulae, blastocysts, and late blastocysts produced in vivo and in vitro. Embryos produced in vivo were obtained from superovulated gilts after flushing the oviduct or uterus. Embryos cultured in vitro were developed from zygotes produced in vivo. Embryos were cultured in vitro to appropiate stages of development in chemically defined medium, North Carolina State University (NCSU)-23. For analysis of the type of lipid in the fat droplets, embryos were fixed in 2.5% glutaraldehyde with the addition of 3 mM calcium chloride. The material was then embedded in Technovit 8100, cut into semi-thin sections, and analyzed by histochemical methods. Four techniques were used to detect different types of lipids: Churukian method with Oil red O, Cain method with Nile blue sulfate, Sudan black B, and osmium tetroxide methods. Fat droplets of embryos produced both in vivo and in vitro contained unsaturated hydrophobic lipids, free fatty acids, phospholipids, unsaturated esters, and triglycerides. Moreover, in the morula the total amount of lipids (especially the amount of free fatty acids and phospholipids) evidently decrease. The amount of the other unsaturated lipids decreased as early as the 2- to 4-cell stage. In conclusion, the content of different types of lipids in pig embryos is reduced during their development from zygote to blastocyst, and there are no differences in lipid composition of fat droplets between in vivo- and in vitro-produced porcine embryos. This research was funded by the State Committee for Scientific Research (Project No. 2 P06D 003 26).

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Degradation of Membrane Phospholipids and Thiols in Peroxide Hemolysis: Studies in Vitamin E Deficiency

Blood ◽

10.1182/blood.v32.4.549.549 ◽

1968 ◽

Vol 32 (4) ◽

pp. 549-568 ◽

Cited By ~ 96

Author(s):

HARRY S. JACOB ◽

SAMUEL E. LUX

Keyword(s):

Fatty Acids ◽

Vitamin E ◽

Hemolytic Anemia ◽

Phosphatidyl Ethanolamine ◽

Red Cells ◽

Membrane Phospholipids ◽

Vitamin E Deficiency ◽

Red Cell Membrane ◽

Inhibition Of Metabolism

Abstract To understand more clearly the hemolytic anemia associated with administration of certain oxidant drugs, the mechanism by which H2O2 causes hemolysis in rat red cells, deficient in vitamin E was investigated. It was demonstrated that the locus of attack by H2O2 was the red cell membrane, in which one phospholipid, i.e., phosphatidyl ethanolamine, was specifically destroyed prior to the onset of hemolysis. No perturbation of intracellular components or metabolism was noted during peroxidative hemolysis. E-deficient red cells incorporated 14C-labelled fatty acids into this phosphatide at nearly twice the rate that in E-supplemented cells, reflecting the continual tendency of phosphatidyl ethanolamine to be destroyed. Young red cells were especially active in this regard and concomitantly were less vulnerable to damage by H2O2 both in vitro and when circulating in rats exposed to hyperbaric oxygenation. If, however, replacement of fatty acids in phosphatidyl ethanolamine was prevented by inhibition of metabolism or if fatty acids were enzymatically removed by a phospholipase-A, H2O2 hemolysis was potentiated. Hemolysis was also associated with, and potentiated by, loss of membrane sulfhydryl activity. It is suggested that hemolytic anemia may occur in patients with vitamin E deficiency (i.e., with steatorrhea) if oxidant drugs capable of generating H2O2 and oxidizing membrane thiols are administered. Two such cases are under investigation.

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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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A novel in vitro model for joint effects of alcohol and free fatty acids on hepatocellular steatosis and inflammation

Zeitschrift für Gastroenterologie ◽

10.1055/s-0032-1331932 ◽

2013 ◽

Vol 51 (01) ◽

Author(s):

A Mahli ◽

WE Thasler ◽

M Müller ◽

C Hellerbrand

Keyword(s):

Fatty Acids ◽

Free Fatty Acids ◽

In Vitro Model ◽

Joint Effects ◽

Vitro Model

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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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