Morphological Changes In Erythrocytes Research Articles

In vitro antiplasmodial effect of ethanolic extracts of traditional medicinal plant Ocimum species against Plasmodium falciparum

Objective To identify the possible antiplasmodial compounds from leaf, stem, root and flower extracts of Ocimum canum (O. canum), Ocimum sanctum (O. sanctum) and Ocimum basilicum (O. basilicum). Methods The O. canum, O. sanctum and O. basilicum were collected from Ramanathapuram District, Tamil Nadu and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 μg/mL) of leaf, stem, root and flower extracts of O. canum, O. sanctum and O. basilicum were tested for antiplasmodial activity against Plasmodium falciparum (P. falciparum). The potential extracts were also tested for their phytochemical constituents. Results The leaf extract of O. sanctum showed excellent antiplasmodial activity (IC 50 35.58 μg/mL) followed by leaf extract of O. basilicum (IC 50 43.81 μg/mL). The leaf extract of O. canum, root extracts of O. sanctum and O. basilicum, the stem and flower extracts of all the three tested Ocimum species showed IC 50 values between 50 and 100 μg/mL. Statistical analysis reveals that, significant antiplasmodial activity ( P <0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it shows that, there were no morphological changes in erythrocytes by the ethanolic extract of O. canum, O. sanctum and O. basilicum. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, flavonoids, phenols, saponins, triterpenoids, proteins, resins, steroids and tannins in the ethanolic extracts of tested plants. Conclusions The ethanolic leaf extracts of O. sanctum possess lead compounds for the development of antiplasmodial drugs.

In vitro antiplasmodial activity of chosen terrestrial medicinal plants against Plasmodium falciparum

ObjectiveTo identify the possible antiplasmodial compounds from Carica papaya (C. papaya), Pongamia glabra (P. glabra), Phyllanthus emblica (P. emblica) L, Phyllanthus acidus (P. acidus) L, Pisonia grandis (P. grandis) and Moringa pterygosperma (M. pterygosperma). MethodsThe C. papaya, P. glabra, P. emblica L, P. acidus L, P. grandis and M. pterygosperma were collected from Ramanathapuram District, Tamil Nadu, India and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 μg/mL) were tested for antiplasmodial activity against Plasmodium falciparum (P. falciparum). The potential extracts were also tested for their phytochemical constituents. ResultsOf the selected plants species, the leaf extract ofP. emblicaL showed excellent antiplasmodial activity (IC50 35.09 μg/mL) followed by leaf extract of M. pterygosperma (IC50 42.36 μg/mL), leaf extract of C. papaya (IC50 46.23 μg/mL) and leaf extract of P. glabra (IC50 48.17 μg/mL). The flower extract of M. pterygosperma showed IC50 values of more than 100 μg/mL. Statistical analysis reveals that, significant antiplasmodial activity (P<0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it shows that, there were no morphological changes in erythrocytes by the ethanolic extract of all the tested plant extracts. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, carbohydrates, flavonoids, phenols, saponins, triterpenoids, steroids and tannins in the ethanolic extracts of tested plants. ConclusionsThe ethanolic stem extracts of P. emblica L possess lead compounds for the development of antiplasmodial drugs.

In vitro antiplasmodial activity of bacterium RJAUTHB 14 associated with marine sponge Haliclona Grant against Plasmodium falciparum

Malaria is the most important parasitic disease, leading to annual death of about one million people, and the Plasmodium falciparum develops resistance to well-established antimalarial drugs. The newest antiplasmodial drug from a marine microorganism helps in addressing this problem. In the present study, Haliclona Grant were collected and subjected for enumeration and isolation of associated bacteria. The count of bacterial isolates was maximum in November 2007 (18 × 10(4) colony-forming units (CFU) g(-1), and the average count was maximum during the monsoon season (117 × 10(3) CFU g(-1)). Thirty-three morphologically different bacterial isolates were isolated from Haliclona Grant, and the extracellular ethyl acetate extracts were screened for antiplasmodial activity against P. falciparum. The antiplasmodial activity of bacterium RJAUTHB 14 (11.98 μg[Symbol: see text]ml(-1)) is highly comparable with the positive control chloroquine (IC(50) 19.59 μg[Symbol: see text]ml(-1)), but the other 21 bacterial extracts showed an IC(50) value of more than 100 μg[Symbol: see text]ml(-1). Statistical analysis reveals that significant in vitro antiplasmodial activity (P < 0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes showed no morphological changes in erythrocytes by the ethyl acetate extract of bacterial isolates after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of reducing sugars and alkaloids in the ethyl acetate extracts of bacterium RJAUTHB 14. The 16S rRNA gene partial sequence of bacterium RJAUTHB 14 is deposited in NCBI (GenBank accession no. GU269569). It is concluded from the present study that the ethyl acetate extracts of bacterium RJAUTHB 14 possess lead compounds for the development of antiplasmodial drugs.

In vitro antiplasmodial activity of marine sponge Hyattella intestinalis associated bacteria against Plasmodium falciparum

Objective To identify the antiplasmodial drugs from the marine sponge Hyattella intestinalis ( H. intestinalis) associated bacteria. Methods The H. intestinalis samples were collected from Thondi coast and subjected for enumeration and isolation of associated bacteria. Filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125μ/mL) from bacterial isolates were screened for antiplasmodial activity against P. falciparum and potential extracts were also screened for biochemical constituents. Results The count of THB isolates were maximum in November 2007 (20×10 4 CFU/g) and the average count was maximum during the monsoon season (77×10 3 CFU/g). A total of 29 bacteria were isolated based on the morphological characteristics and screened for antiplasmodial activity. The antiplasmodial activity of THB20 extract (IC 50 41.88 μ/mL) showed at two fold concentration of IC 50 value of the positive control chloroquine (IC 50 19.59 μ/mL) and 14 bacterial isolates showed IC 50 value of more than 100 μ/mL. Statistical analysis reveals that, significant in vitro antiplasmodial activity ( P<0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes showed no morphological changes in erythrocytes by the ethyl acetate extract of THB isolates after 48 h of incubation. The antiplasmodial activity of potential bacterial isolates might be due to the presence of sugars and alkaloids in the ethyl acetate extracts. Conclusions It is concluded from the present study that, the ethyl acetate extracts of THB20 posses novel metabolites for the development of newer antiplasmodial drugs.

Regulation of membrane-cytoskeletal interactions by tyrosine phosphorylation of erythrocyte band 3

AbstractThe cytoplasmic domain of band 3 serves as a center of erythrocyte membrane organization and constitutes the major substrate of erythrocyte tyrosine kinases. Tyrosine phosphorylation of band 3 is induced by several physiologic stimuli, including malaria parasite invasion, cell shrinkage, normal cell aging, and oxidant stress (thalassemias, sickle cell disease, glucose-6-phosphate dehydrogenase deficiency, etc). In an effort to characterize the biologic sequelae of band 3 tyrosine phosphorylation, we looked for changes in the polypeptide's function that accompany its phosphorylation. We report that tyrosine phosphorylation promotes dissociation of band 3 from the spectrin-actin skeleton as evidenced by: (1) a decrease in ankyrin affinity in direct binding studies, (2) an increase in detergent extractability of band 3 from ghosts, (3) a rise in band 3 cross-linkability by bis-sulfosuccinimidyl-suberate, (4) significant changes in erythrocyte morphology, and (5) elevation of the rate of band 3 diffusion in intact cells. Because release of band 3 from its ankyrin and adducin linkages to the cytoskeleton can facilitate changes in multiple membrane properties, tyrosine phosphorylation of band 3 is argued to enable adaptive changes in erythrocyte biology that permit the cell to respond to the above stresses.

Seaweeds as a source of lead compounds for the development of new antiplasmodial drugs from South East coast of India

The problems of resistant lines of Plasmodium falciparum are escalating. Twelve seaweeds species belong to five different families (Sargassaceae, Gracilariaceae, Hypneaceae, Corallinaceae and Halimedaceae) were collected from Mandapam coastal area, and the seaweeds extracts were tested for in vitro antiplasmodial activity against P. falciparum. Among the tested seaweeds, Gracilaria verrucosa (IC(50) 5.55 μg.ml(-1)) and Hypnea espera (IC(50) 8.94 μg.ml(-1)) showed good antiplasmodial activity, and these results are comparable with positive controls such as artemether (IC(50) 4.09 μg.ml(-1)) and chloroquine (IC(50) 19.59 μg.ml(-1)), respectively. Turbinaria conoides, Sargassum myriocystem, Hypnea valentiae and Jania rubens extracts showed IC(50) values between 5 to 50 μg.ml(-1). Sargassum sp., Turbinaria decurrens and Halimeda gracilis extracts showed IC(50) values between 50 to 100 μg.ml(-1). Gracilaria corticata, Jania adherens and Halimeda opuntia extracts showed IC(50) value of more than 100 μg.ml(-1). Statistical analysis reveals that significant in vitro antiplasmodial activity (P < 0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out, and it shows that no morphological changes in erythrocytes by the ethanolic extract of seaweeds extracts after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of sugars, proteins, phenols and carboxylic acid in the ethanolic extracts of seaweeds. It is concluded from the present study that the ethanolic extracts of seaweeds of G. verrucosa and Hypnea espera possess lead compounds for development of antiplasmodial drugs.

In vitro antiplasmodial activity of ethanolic extracts of seaweed macroalgae against Plasmodium falciparum

Malaria is a major health problem in many developing countries. The drugs resistant Plasmodium falciparum causes the most virulent form of malaria in humans and it is described as a public health disaster causing increased morbidity and mortality. Thirteen seaweeds species which belong to four different families (Rhodomelaceae, Cladophoraceae, Ulvaceae, and Caulerpaceae) were collected from Mandapam coastal area and the seaweeds extracts were tested for in vitro antiplasmodial activity against P. falciparum. Among them, Caulerpa toxifolia (IC(50) 5.06 μg·ml(-1)) showed potential antiplasmodial activity than other seaweeds extracts and it can be comparable with the positive control artemether (IC(50) 4.09 μg·ml(-1)). Caulerpa peltata (IC(50) 16.69 μg·ml(-1)) also exhibited good antiplasmodial activity and the IC(50) value is lesser than the positive control chloroquine (IC(50) 19.59 μg·ml(-1)). Statistical analysis reveals that significant in vitro antiplasmodial activity (P<0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it shows that no morphological changes in erythrocytes by the ethanolic extract of seaweeds extracts after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of sugars, proteins, and phenols in the ethanolic extracts of seaweeds. It is concluded from the present study that, the ethanolic extracts of seaweeds of C. toxifolia and C. peltata possesses lead compounds for development of antiplasmodial drugs.

Mangrove plants as a source of lead compounds for the development of new antiplasmodial drugs from South East coast of India

Malaria is the world's leading killer among the infectious diseases. The treatment of malaria is mystified by the challenges of widespread resistance of the malaria parasites to cheap and affordable antimalarial drugs. The present study was made in an attempt to identify the in vitro antiplasmodial activity against mangrove plant parts. (Avicennia marina, Acanthus ilicifolius, Bruguiera cylindrica, Excoecaria agallocha, Rhizophora apiculata, and Rhizophora mucronata mangrove plant extracts exhibited in vitro antiplasmodial activity against Plasmodium falciparum). Of the selected mangrove plant parts, the bark extract of A. marina exhibited minimum concentration of inhibitory activities IC(50) 49.63 μg.ml(-1). The leaf extract of A. marina, the hypocotyl extract of B. cylindrica, the leaf extract of E. agallocha, the flower extract of R. mucronata, and the hypocotyl extract of R. apiculata showed IC(50) values between 50 and 100 μg.ml(-1). Statistical analysis reveals that significant antiplasmodial activity (P<0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out, and it shows that there were no morphological changes in erythrocytes by the ethanolic extract of mangrove plants after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of alkaloids, carboxylic acids, coumarins, saponins, flavonoids, xanthoproteins, tannins, phenols, sugars, resins, steroids, and proteins in the ethanolic extracts of mangrove plants. It is concluded from the present study that the ethanolic extracts of mangrove plant parts of A. marina possess lead compounds for the development of antiplasmodial drugs.

UP-1.091: Plastic Orchiectomy and the Morphological Changes of Erythrocytes

It is known that erythrocyte defense function that is absorption, transportation and participation in metabolic processes, in norm is regulative mechanism but its alteration plays role in pathogenesis of cancer diseases. The aim of the work was to study erythrocyte morphology by means of electron microscopy in given pathology.

The interaction of DNR and glutaraldehyde with cell membrane proteins leads to morphological changes in erythrocytes

In this study, the effects of DNR and glutaraldehyde on isolated erythrocyte membrane proteins were examined. For this purpose, SDS–gel electrophoresis was carried out. Additionally, analyses of the disturbances in erythrocyte shape and size, accompanied by the application of flow cytometry and microscopy examination, were undertaken. The amount of DNR linked to erythrocyte cell membranes was measured by a fluorimetric technique. It was observed that glutaraldehyde caused in concentration dependent manner an increase of percent of DNR linked to cell membrane proteins. After this incorporation, perturbations in the protein content of cell membranes were observed. The protein aggregates and changes in the level of spectrin, actin and band 3 protein were noted. Due to the changes in spectrin, which is mainly responsible for maintenance of the discocyte shape of erythrocytes, flow cytometry and microscopy techniques were used to control the size and shape of erythrocytes after treatment with DNR and glutaraldehyde. The disturbances in the shape and size of erythrocytes were observed for all tested concentrations of glutaraldehyde. For all tested concentrations of glutaraldehyde, the changes were statistically significant.

Ontogenetic changes in erythrocyte morphology in larval mole salamanders, Ambystoma talpoideum, measured with image analysis

Knowledge of the erythrocyte morphology in an animal can provide information regarding its genome size, metabolic rate, exposure to pollutants and acclimatization to its environment. In amphibians, there are thought to be two morphological forms of erythrocytes, a larger, elongated larval type and a smaller, rounder adult form. I examined erythrocyte morphology in larval and paedomorphic adult mole salamanders, Ambystoma talpoideum using an image analysis approach to measure dimensions of 30,213 erythrocytes in 66 individuals captured from a single pond. The erythrocytes of this species averaged 590.3 μm2 in area, 34.7 μm in length, and 22.6 μm in width, which is comparable to values reported from related Ambystomatid species. There was a tendency for cells to decrease in length with increasing salamander size, and a stronger positive relationship between body size and erythrocyte width, so that cells were more rounded overall in larger individuals. There was also less variability in certain erythrocyte dimensions in larger individuals. However, cells did not decrease in area as salamanders grow, contrary to expectations. I conclude with comments on the image analysis approach used in this study, which proved especially useful in gathering this data, as it allowed for large sample sizes, direct measurement of surface areas (of one side of cells), and was nearly fully automated.

Cambios de la morfología de los eritrocitos inducidos por clorpromazina e imipramina

Chlorpromazine (CPZ), a phenothiazine derivative, is a potent antipsychotic agent and imipramine (IP) is a widely used tricyclic antidepressant. The interaction between these molecules and erythrocyte membranes is of particular interest considering the role of these cells in the transport and release of these drugs at the central nervous system. In the present paper, we intend to study the effects of IP on erythrocyte membranes and to compare these effects with those of CPZ. Erythrocytes from adult Sprague-Dawley rats were incubated separately with different concentrations of IP or CPZ for lh at room temperature, fixed and stained by Giemsa. Changes in erythrocyte morphology were quantified by an image analysis system. The interaction of both drugs, CPZ and IP, with the erythrocyte membrane causes similar changes in cell shape. Increasing concentrations of both drugs induces the formation of stomatocytes, spherostomatocytes and spherocytes, because of an irreversible loss of area and volume, probably due to endovesiculation. Our results also show that the CPZ is more potent than IP.

Bordetella adenylate cyclase toxin induces a cascade of morphological changes of sheep erythrocytes and localizes into clusters in erythrocyte membranes

Adenylate cyclase toxin (CyaA) of Bordetella pertussis penetrates the membrane of eukaryotic cells, producing high levels of intracellular cAMP, as well as hemolysis that results from the formation of cation-selective toxin channels in the membrane. Using several microscopical approaches we studied the effects of CyaA action on the morphology of sheep erythrocytes during early phases preceding lysis and examined localization of CyaA molecules within the erythrocyte membrane. CyaA induced a cascade of morphological changes of erythrocytes, such as shrinkage, formation of membrane projections, and blebs and swelling. The use of an enzymatically inactive CyaA-AC- toxoid that is unable to produce cAMP and of a CyaA-E581K mutant exhibiting higher hemolytic activity than with CyaA showed that the hemolytic activity is responsible for the induction of morphological changes of erythrocytes. Further, immunolabeling of inserted CyaA-232/FLAG molecules with specific anti-FLAG antibodies and IgG-gold particles indicated a clustered distribution of CyaA molecules in erythrocyte membrane. This was confirmed by immunofluorescence and confocal microscopy, which revealed uniform stoichiometry of CyaA clusters, suggesting CyaA binding into specific domains in erythrocyte membrane. Indeed, a decrease of CyaA binding after cholesterol depletion of erythrocytes suggests toxin targeting and binding to membrane microdomains (rafts).

Antioxidant, pro‐oxidant effect of the thiosemicarbazone derivative Schiff base (4‐(1‐phenylmethylcyclobutane‐3‐yl)‐2‐(2‐hydroxybenzylidenehydrazino) thiazole) and its metal complexes on rats

Adverse biological activities of thiosemicarbazone (TSC) and Schiff base (SB) derivatives have been widely studied in rats and in other animal species using different doses, times and routes of administration. However, there are few studies describing changes in some biochemical parameters in vivo which are indicative of oxidative stress in biological systems and of morphological changes of tissues. In this study, the rats were injected subcutaneously with a new thiosemicarbazone thiazole ring containing a Schiff base (LH) and its Cu(L)2 and Zn(L)2 complexes (25 mg kg(-1) body weight) and then sacrificed after 1, 2, 4, 8, 16, 32 and 64 days. The aim of this study was to determine the effect of the new compounds on the serum antioxidant vitamins (A, E, C), selenium (Se), malondialdehyde (MDA) levels, erythrocyte GSH-Px enzyme activity and morphological changes in the liver, kidney and adrenal gland tissues. It was observed that erythrocyte GSH-Px activity, serum MDA and vitamins A, E concentrations were statistically changed (p < 0.02), but serum levels of selenium, and vitamin C were not changed. In conclusion, the parameters measured show that Cu(L)2 caused considerable oxidative stress and Zn(L)2 behaved as an antioxidant. No oxidative stress in LH was observed compared to the control group.

Effects of gallium on common carp ( Cyprinus carpio): acute test, serum biochemistry, and erythrocyte morphology

Gallium (Ga) is one of the intermetallic elements that are increasingly being used in making high-speed semiconductors such as gallium arsenide. The purposes of this study were to investigate the effects of gallium on acute toxicity, on serum biochemical variables as well as on erythrocyte morphological changes in the blood stream of common carp ( Cyprinus carpio). Median lethal concentrations were determined in acute tests. The 96-h LC 50 value was 19.78 (18.49–21.16) mg l −1. Common carp were exposed to different gallium concentrations (2.0, 4.0, and 8.0 mg l −1) for 28 days in laboratory toxicity tests. Means of the measured serum biochemistry parameters (including glucose, blood urea nitrogen, creatinine, cholesterol, and triglyceride) of these exposed groups significantly differed from those of the untreated group. Deformation of erythrocytes suggest disturbance of respiration as an additional indicator of Ga exposure. Our results suggest that 2.0 mg l −1 is proposed as a biologically safe concentration which can be used for establishing tentative water quality criteria concerning of same size common carp. In addition, serum biochemical parameters as well as erythrocyte morphological changes are promising clinical diagnostic tools for assessing the effects of gallium compounds on common carp.

The effect of lead ions on the energy metabolism of human erythrocytes in vitro

The aim of this work was to evaluate the influence of chronic exposure to lead ions on the parameters of energetic status of human erythrocytes in vitro. Umbilical cord erythrocytes were incubated with lead acetate at final lead ion concentrations ranging from 10 to 200 μg/dl. ATP, ADP, AMP, adenosine, GTP, GDP, GMP, guanosine, IMP, inosine, hypoxanthine, NAD and NADP concentrations in erythrocytes were determined using HPLC. Scanning electron micrographs of erythrocytes were taken. The mean concentrations of ATP, GTP, NAD and NADP, and mean values of adenylate energy charge (AEC) and GEC in cells incubated at the presence of lead ions were significantly lower after 20 h of incubation. Concentrations of purine degradation products (Ado, Guo, Ino) and Hyp were significantly higher. It is suggested that lead ions affect the energy metabolism of erythrocytes. Morphological changes in erythrocytes correspond to the increase of lead ions in the incubation mixture and to the decrease of ATP concentration in erythrocytes. A decrease in NAD and ATP concentration in erythrocytes could be a sensitive indicator of energy process disturbance, useful in monitoring in case of chronic lead exposure.

Primaquine-induced hemolytic anemia: effect of 6-methoxy-8-hydroxylaminoquinoline on rat erythrocyte sulfhydryl status, membrane lipids, cytoskeletal proteins, and morphology.

Previous studies have shown that 6-methoxy-8-hydroxylaminoquinoline (MAQ-NOH), an N-hydroxy metabolite of the antimalarial drug, primaquine, is a direct-acting hemolytic agent in rats. To investigate the mechanism underlying this hemolytic activity, the effects of hemotoxic concentrations of MAQ-NOH on rat erythrocyte sulfhydryl status, membrane lipids, skeletal proteins, and morphology have been examined. Treatment of rat erythrocytes with a TC(50) concentration of MAQ-NOH (350 microM) caused only a modest and transient depletion of reduced glutathione (GSH) (~30%), which was matched by modest increases in the levels of glutathione disulfide and glutathione-protein mixed disulfides. Lipid peroxidation, as measured by thiobarbituric acid-reactive substances and F(2)-isoprostane formation, was induced in a concentration-dependent manner by MAQ-NOH. However, the formation of disulfide-linked hemoglobin adducts on membrane skeletal proteins and changes in erythrocyte morphology were not observed. These data suggest that hemolytic activity results from peroxidative damage to the lipid of the red cell membrane and is not dependent on skeletal protein thiol oxidation. However, when red cell GSH was depleted (>90%) by titration with diethyl maleate, hemolytic activity of MAQ-NOH was markedly enhanced. Of interest, exacerbation of hemotoxicity was not matched by increases in lipid peroxidation, but by the appearance of hemoglobin-skeletal protein adducts. Collectively, the data are consistent with the concept that MAQ-NOH may operate by more than one mechanism; one that involves lipid peroxidation in the presence of normal amounts of erythrocytic GSH, and one that involves protein oxidation in red cells with low levels of GSH, such as are seen in individuals with glucose-6-phosphate dehydrogenase deficiency.

Human erythroid cells are affected by aluminium. Alteration of membrane band 3 protein

There is evidence that anaemia is associated with aluminium (Al). We have already reported on the sensitivity to Al, showed by erythroid cell populations of animals chronically exposed to the metal. In order to investigate whether Al could also affect human cells, experiments were carried out both on immature and mature human erythroid cells. Erythroid progenitors (CFU-E, colony-forming units-erythroid) concentrated from human peripheral blood were cultured in an Al-rich medium under erythropoietin stimulation and their development analysed. Human peripheral erythrocytes were aged in the presence of Al. Cells were examined using scanning electron microscopy, and membrane proteins analysed by polyacrylamide gel electrophoresis with sodium dodecyl sulphate and immunoblotting. The development of the Al-treated progenitors was 8750/6600–9200 CFU-E/10 6 cells, a significantly lower median value ( P<0.05) than that showed by non-treated cells (12 300/11 200–20 700 CFU-E/10 6 cells). Erythrocyte morphological changes were induced by Al during the in vitro ageing. The cells lost their typical biconcave shape, turning into acanthocytes and stomatocytes. Simultaneously, an increased membrane protein breakdown compatible with band 3 degradation was detected. Besides, Al was found within the cells and attached to the membrane. The present in vitro results suggest that Al may disturb human erythropoiesis through combined effects on mature erythrocytes and cellular metabolism in late erythroid progenitors.

Effect of bilirubin on toxicity induced by trifluoperazine, dibucaine and praziquantel to erythrocytes

Unconjugated bilirubin (UCB), like trifluoperazine (TFP), dibucaine (DBC) and praziquantel (PZQ), induces erythrocyte morphological changes, lysis and lipid exfoliation. In the present study we determined whether TFP, DBC and PZQ toxicity to erythrocytes was potentiated or reverted by UCB. Human erythrocytes were either treated or non-treated with 34.2 μmol/L UCB for 10 min prior to the incubation with toxic concentrations of TFP (0.12 mmol/L), DBC (1.5 mmol/L) or PZQ (3.0 mmol/L), for 1 h (37°C). Studies of toxic effects included morphological analysis of erythrocytes, evaluation of hemoglobin release and loss of membrane lipids. Although UCB has an echinocytogenic effect, its co-incubation with TFP or PZQ did not alter the stomatocytogenic effect of the drug but enhanced DBC-induced stomatocytosis. Cell fusion was a common feature in experiments with DBC. Injurious effect of DBC to erythrocytes was potentiated by UCB as manifested by a marked increase in hemolysis (171%, p<0.05), and in elution of membrane cholesterol (73%, p<0.01) and phospholipids (123%, p<0.01). In opposite, toxic events produced by TFP and PZQ to erythrocytes were not aggravated by UCB. Interestingly, UCB prevented the loss of membrane cholesterol by PZQ (−36%, p<0.01), as well as that of phospholipids by TFP (−28%, p<0.05). These findings indicate that UCB potentiates DBC injury to erythrocytes, while protects membrane lipid elution by PZQ and TFP. Therefore, the relation of the benefits and risks of the administration of DBC to jaundiced patients should be carefully considered.

Scanning electron microscopy of rat erythrocytes during chronic alcoholic intoxication combined with protein and vitamin deficit.

Electron microscopy revealed morphological changes in erythrocytes from rats with chronic alcoholic intoxication kept on protein- and vitamin B-deficient rations. All animals had anisopoikilocytosis (up to 50%). Most pronounced changes in erythrocyte population attesting to accelerated erythrocyte aging (stomato- and microcytosis, discocyte swelling, and spontaneous hemolysis) were found in alcohol-fed rats kept on deficient ration.