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Uncaria tomentosa (Willd.) DC is a woody climber species originating from South and Central America that has been used in the therapy of asthma, rheumatism, hypertension, and blood purification. Our previous study showed that U. tomentosa extracts altered human erythrocyte shape, which could be due to incorporation of the compounds contained in extracts into the erythrocyte membrane. The aim of the present study was to determine how the compounds contained in U. tomentosa extracts incorporate into the human erythrocyte membrane. The study has assessed the effect of aqueous and ethanolic extracts from leaves and bark of U. tomentosa on the osmotic resistance of the human erythrocyte, the viscosity of erythrocyte interior, and the fluidity of erythrocyte plasma membrane. Human erythrocytes were incubated with the studied extracts in the concentrations of 100, 250, and 500 µg/mL for 2, 5, and 24 h. All extracts tested caused a decrease in erythrocyte membrane fluidity and increased erythrocyte osmotic sensitivity. The ethanolic extracts from the bark and leaves increased viscosity of the erythrocytes. The largest changes in the studied parameters were observed in the cells incubated with bark ethanolic extract. We consider that the compounds from U. tomentosa extracts mainly build into the outer, hydrophilic monolayer of the erythrocyte membrane, thus protecting the erythrocytes against the adverse effects of oxidative stress.

Numerous research works have shown that synthesis of pesticides leads to the formation of impurities that may substantially enhance pesticide toxicity. In this study, the effect of manufacturing impurities of pesticide bromfenvinphos (BFVF) such as 1-bromo-2-(2,4-dichlorophenyl)-2-ethoxy ethene (BDCEE) and diethyl [2-(2,4-dichlorophenyl)-2-oxo-ethyl] phosphonate (β-ketophosphonate) on human erythrocytes, being significantly exposed to xenobiotics has been studied. The cells were treated with the compounds studied in the concentrations ranging from 0.1 μM to 250 μM for 4 h. In order to assess the effect of BDCEE and β-ketophosphonate on red blood cells hemolytic changes, changes in cell size (FSC parameter) and oxidation of hemoglobin were studied. Moreover, alterations in reactive oxygen species (ROS) formation, reduced glutathione (GSH) level and acetylcholinesterase (AChE) activity were determined. BDCEE induced an increase in ROS level and caused strong oxidation of hemoglobin as well as a slight change in erythrocytes size and hemolysis, while it did not change GSH level and AChE activity. β-ketophosphonate has not been shown to affect most parameters studied, but it strongly reduced AChE activity. Because changes in the parameters examined were noted at low concentrations of BFVF impurities (5-250 µM), those substances should not negatively affect on red blood cells of humans environmentally exposed to this pesticide.

Phenol and chlorinated phenols are widely spread in the environment and human surrounding, which leads to a common environmental and occupational exposure of humans to these substances. The aim of this study was to assess eryptotic changes in human red blood cells treated with phenol, 2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP) and pentachlorophenol (PCP). The erythrocytes were incubated with phenols studied in the concentrations ranging from 1 to 100 μg/mL for 24 h or 48 h. The results of the study revealed that all compounds studied caused phosphatidylserine translocation and increased cytosolic calcium ions level in human erythrocytes. It was also noticed that phenol and chlorophenols caused an increase in caspase-3 and calpain activation, which confirmed that they were capable of inducing suicidal death of erythrocytes. The results also revealed that PCP most strongly altered the parameters studied, while phenol exhibited the weakest eryptotic potential in the incubated cells., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) and its analogs are widely used in the production of various everyday use products, which leads to a common exposure of humans to these substances. The effect of bisphenols on oxidative stress parameters has not been described in detail in non-nucleated cells, therefore, we have decided to evaluate the impact of BPA and its analogs, i.e. bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF) on reactive oxygen species (ROS) formation, lipid peroxidation, glutathione (GSH) level and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in human erythrocytes. The erythrocytes were incubated with the compounds studied in the concentrations ranging from 0.1 to 500μg/ml for 1, 4 or 24h. It has been found that bisphenols enhanced ROS (including • OH) formation, depleted GSH level, increased lipid peroxidation and changed the activities of SOD, CAT and GSH-Px. It has been noted that the strongest alterations in ROS formation, lipid peroxidation and the activity of antioxidant enzymes were induced by BPAF, which changed CAT and SOD activity even at 0.5μg/ml. It has also been shown that BPA caused the strongest changes in GSH level, while BPS, which is the main BPA substituent in the manufacture did not alter most parameters studied., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Bisphenols are important chemicals that are widely used in the manufacturing of polycarbonates, epoxy resin and thermal paper, and thus the exposure of humans to these substances has been noted. The purpose of this study was to assess eryptotic changes in human erythrocytes exposed (in vitro) to bisphenol A (BPA) and its selected analogs, i.e.,bisphenol F (BPF), bisphenol S (BPS) and bisphenol AF (BPAF). The erythrocytes were incubated with compounds studied at concentrations ranging from 1 to 250μg/mL for 4, 12 or 24h. The results showed that BPA and its analogs increased cytosolic calcium ions level with the strongest effect noted for BPAF. It has also been revealed that all bisphenols analyzed, and BPAF and BPF in particular increased phosphatidylserine translocation in red blood cells, which confirmed that they exhibited eryptotic potential in this cell type. Furthermore, it was shown that BPA and its analogs caused significant increase in calpain and caspase-3 activities, while the strongest effect was noted for BPAF. BPS, which is the main substituent of bisphenol A in polymers and thermal paper production exhibited similar eryptotic potential to BPA. Eryptotic changes in human erythrocytes were provoked by bisphenols at concentrations, which may influence the human body during occupational exposure or subacute poisoning with these compounds., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) has been shown to provoke many deleterious impacts on human health, and thus it is now successively substituted by BPA analogues, whose effects have been poorly investigated. Up to now, only one study has been realized to assess the effect of BPA on human erythrocytes, which showed its significant hemolytic and oxidative potential. Moreover, no study has been conducted to evaluate the effect of BPA analogues on red blood cells. The purpose of the present study was to compare the impact of BPA and its selected analogues such as bisphenol F (BPF), bisphenol S (BPS) and bisphenol AF (BPAF) on hemolytic and morphological changes and hemoglobin oxidation (methemoglobin formation) of human erythrocytes. The erythrocytes were incubated with different bisphenols concentrations ranging from 0.5 to 500μg/ml for 1, 4 and 24h. The compounds examined caused hemolysis in human erythrocytes with BPAF exhibiting the strongest effect. All bisphenols examined caused methemoglobin formation with BPA inducing the strongest oxidative potential. Flow cytometry analysis showed that all bisphenols (excluding BPS) induced significant changes in erythrocytes size. Changes in red blood cells shape were conducted using phase contrast microscopy. It was noticed that BPA and BPAF induced echinocytosis, BPF caused stomatocytosis, while BPS did not provoke significant changes in shape of red blood cells. Generally, the results showed that BPS, which is the main substituent of bisphenol A in polymers and thermal paper production, exhibited significantly lower disturbance of erythrocyte functions than BPA., (Copyright © 2015 Elsevier Inc. All rights reserved.)

In this study, we continued our investigations concerning the interaction of Uncaria tomentosa extracts with the human erythrocytes. The analysis of the size and shape of the erythrocytes by means of flow cytometry and phase contrast microscopy was performed. We executed our experiments using ethanolic and aqueous extracts from the leaves and bark of U. tomentosa. Disturbances were observed in the size and shape of the erythrocytes incubated with ethanolic and aqueous extracts at the concentrations of 100 μg/mL and 250 μg/mL, respectively. The observed changes were probably related to the entry of polyphenolic compounds contained in U. tomentosa extracts into erythrocyte membrane. Externalization of phosphatidylserine on the erythrocytic surfaces was also noticed during incubation with extracts at concentration of 250 μg/mL. We concluded that all of the extracts examined induced changes in the erythrocyte membrane properties, whereas ethanolic extracts from bark induced the most significant changes. The possible binding of polyphenols to the erythrocyte surface may have accounted for the protective properties of extracts against haemolysis of RBCs, which was observed in our previous study (Bors et al., 2011), but considerable incorporation of polyphenols into cell membranes can result in disturbance of phosphatidylserine transport and changes in erythrocyte shape. Nevertheless the results of the investigations showed that considerable morphological changes appear only as a result of erythrocyte exposure to high concentrations (50 ppm and 100 ppm) of the extracts studied, thus they should not lead to clinical erythrocytic damage if recommended doses of U. tomentosa preparations are administrated., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The purpose of this study was to evaluate the effect of the ethanolic and aqueous extracts of Uncaria tomentosa on human erythrocytes and additionally the assessment of protective effect of these extracts on hemolysis induction, hemoglobin oxidation, and changes in the level of reactive oxygen species (ROS) and lipid peroxidation, which were provoked by selected xenobiotics, i.e. 2,4-dichlorophenol (2,4-DCP) and catechol. All tested extracts, even at a very high concentration of 500 μg/ml were not toxic to the erythrocytes because they did not cause lipid peroxidation, increase methemoglobin and ROS levels nor provoked hemolysis. The results of this study also revealed protective effect of extracts of U. tomentosa. The extracts studied depleted the extent of hemoglobin oxidation and lipid peroxidation as well as decreased the level of ROS and hemolysis, which was provoked by 2,4-DCP. No protective activity of the extracts against catechol action, which is a precursor of semiquinones in cell was found. A difference in the effect of the extracts studied was observed. Ethanol-based extracts revealed more pronounced ability to inhibit oxidation processes in human erythrocytes., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Although it is known that phenoxyacetic herbicides significantly affect the oxidative status of human erythrocytes, there is no direct evidence of their ability to induce free radical production. To demonstrate this phenomenon we investigated the effect of two commonly used phenoxyherbicides-sodium salt of 2,4-dichlorophenoxyacetic acid (2,4-D-Na) and sodium salt of 4-chloro-2-methylphenoxyacetic acid (MCPA-Na) on oxidation of dihydrorhodamine 123 and H(2)DCFDA as well as on carbonyl group content in cellular proteins. Moreover, haemoglobin denaturation was also measured. The rate of fluorescent probe oxidation was significantly higher for 2,4-D-Na, while both compounds exerted similar effects on protein carbonyl group (an increase in their content) and on denaturation of haemoglobin (no changes were observed). These results and the previous data led us to a conclusion that pro-oxidative action of phenoxyherbicides is strongly dependent on the localization of the substituent in the phenol ring. We also proposed a metabolic reaction chain that explains the mechanism of action of 2,4-D-Na in vivo.

3-(Dimethylamino)phenol (3-DMAP) exists in the environment as a transformation product of ureic herbicides and may also be considered as a derivative of phenoxyherbicides. In this study, the activity of glutathione peroxidase, catalase and superoxide dismutase, as well as the level of free radicals and changes in cell morphology were measured in human erythrocytes exposed (in vitro) to 3-(dimethylamino)phenol. Human erythrocytes were incubated for 1 h in 3-DMAP at concentrations of 10-500 microg per 1 ml erythrocytes of 5% haematocrit. The results show that 3-(dimethylamino)phenol increased the level of free radicals and changed the activity of glutathione peroxidase, catalase, superoxide dismutase and acetylcholinesterase. It also changed cell morphology. All these results corroborated the thesis that 3-DMAP induces oxidative stress in cells. 3-DMAP changed the properties of the cell membrane, caused strong oxidation of haemoglobin, inhibited the levels of enzymatic and non-enzymatic antioxidants, which, in result, lead to generation of free radicals (ROS and semiquinones) that occurred in the exposed cells, predisposing them to oxidative damage.

The effect of phenolic compounds: phenol, 2,4-dichlorophenol (2,4-DCP), 2,4-dimethylphenol (2,4-DMP) and catechol on human erythrocytes was studied. The level of fluorescent label - 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFDA) oxidation by phenolic compounds in erythrocytes as well as the carbonyl group content and hemoglobin denaturation were monitored. H(2)DCFDA has been utilized extensively as a marker for studies of oxidative stress at the cellular level. We noted that 2,4-DCP, 2,4-DMP and catechol induced an increase in the concentration- and time-dependent H(2)DCFDA oxidation. We also observed an increase in carbonyl group content and the changes in parameter T (denaturation of hemoglobin) in erythrocytes incubated with 2,4-DCP, catechol and 2,4-DMP. The highest level of H(2)DCFDA oxidation was provoked by 2,4-DCP. The biggest changes of proteins in erythrocytes measured as the carbonyl group content were induced by 2,4-DMP, but measured as parameter T they were induced by catechol. It was observed that phenol did not oxidize H(2)DCFDA up to the concentration of 2.5 mM after 3 h of incubation. Phenol did not affect the carbonyl group content but decreased parameter T (induced denaturation of hemoglobin). To sum up, the kind of the substituent in a phenolic ring determines the molecular mechanism of action of the individual compound and the capacity of reactive oxygen species generation and thus damages the specified structures in human erythrocytes.

The effect of the exposure of human erythrocytes to different concentrations of microcystin-LR were studied. Lipid peroxidation, membrane fluidity, cell morphology, haemoglobin oxidation and changes in the activity of antioxidant enzymes were investigated. Human erythrocytes were incubated with microcystin-LR at concentrations of 1-1000 nM for 1, 6, 12 and 24 h. We observed that microcystin-LR induces a significant increase of the level of thiobarbituric acid reactive substances (TBARS), formation of echinocytes, haemolysis, conversion of oxyhaemoglobin to methaemoglobin, decrease of membrane fluidity on the level of 16 carbon atom fat acids. The compound also changed antioxidative enzymes activities: catalase, superoxide dismutase and glutathione reductase and formation of reactive oxygen species (ROS). All of the observed changes point out that 100 nM of Microcistin LR is the liminal (threshold) toxic dose for human erythrocytes. This dose caused most of the described changes. Observed damages of erythrocytes membrane and antioxidative enzymes may be the result of direct covalent binding of microcystin-LR with -SH residues of proteins and indirectly be related with reactive oxygen species formation.

The effectiveness of penetration of erythrocyte membrane by sodium salt of 2,4-dichlorophenoxyacetic acid was analyzed. The experiment was executed in a dependence on different doses of the herbicide and at different times of incubation of red blood cells with 2,4-D-Na. It is known that the main mechanism of detoxification of the cell from xenobiotics including 2,4-D is to bind them with proteins contained in blood plasma. In the case of exposure of blood to high doses of 2,4-D-Na, the unbound part of xenobiotics may penetrate into erythrocytes and change the activity of numerous parameters of the cells. The results obtained by the use of high pressure liquid chromatography (HPLC) revealed that 2,4-D-Na is adsorbed by erythrocytes of 5% haematocrite in the amount of ∼1% of the initial concentration. Moreover, it was observed that 2,4-D-Na is capable of accumulating in erythrocyte's membrane and haemolysate in the amounts of 0.15% and 1.23% of the initial concentration, respectively. It was also stated that penetration of 2,4-dichlorophenoxyacetic acid into erythrocytes is not associated with incubation time (the similar concentrations of 2,4-D-Na were detected after different incubation times of 0.5 to 3 hours); however, it was related with concentrations of the herbicide. We suggest that 2,4-D-Na was transported with concentration gradient in human erythrocytes. [ABSTRACT FROM AUTHOR]