Cannabidiol protects keratinocyte cell membranes following exposure to UVB and hydrogen peroxide.

Keratinocytes, the major cell type of the epidermis, are particularly sensitive to environmental factors including exposure to sunlight and chemical agents. Since oxidative stress may arise as a result of these factors, compounds are actively sought that can act as protective agents. Recently, cannabidiol (CBD), a phytocannabinoid found in Cannabis Sativa L., has gained increased interest due to its anti-inflammatory and antioxidant properties, and absence of psychoactive effects. This prompted us to analyze the protective effects of CBD on keratinocytes exposed to UVB irradiation and hydrogen peroxide. Here we show, using liquid chromatography mass spectrometry, that CBD was able to penetrate keratinocytes, and accumulated within the cellular membrane. CBD reduced redox balance shift, towards oxidative stress, caused by exposure UVB/hydrogen peroxide, estimated by superoxide anion radical generation and total antioxidant status and consequently lipid peroxidation level. CBD was found to protect keratinocytes by preventing changes in the composition of the cellular membrane, associated with UVB/hydrogen peroxide damages which included reduced polyunsaturated fatty acid levels, increased sialic acid and lipid peroxidation products (malondialdehyde and 8-isoprostanes) levels. This maintains cell membranes integrity and prevents the release of lactate dehydrogenase. In addition, CBD prevented UVB/hydrogen peroxide-induced reduction of keratinocyte size and zeta potential, and also decreased activity of ATP-binding cassette membrane transporters. Together, these findings suggest that CBD could be a potential protective agent for keratinocytes against the harmful effects of irradiation and chemical environmental factors that cause oxidative stress.

Phospholipidomic Analysis Reveals Changes in Sphingomyelin and Lysophosphatidylcholine Profiles in Plasma from Patients with Neuroborreliosis.

In recent years, the number of patients suffering from Lyme Disease (LD) has significantly increased. The most dangerous manifestation of LD is neuroborreliosis associated with invasion of the central nervous system by Borrelia burgdorferi. Phospholipids (PL) and their metabolites are involved in inflammation, which plays a dominant, but still unclear, role in the pathogenesis of neuroborreliosis. We analyzed the plasma PL profiles of neuroborreliosis patients (n = 8) and healthy volunteers (n = 8) using a lipidomic approach. Significant increases in the lysophosphatidylcholines LysoPtdCho 16:0 and LysoPtdCho 18:2 were observed. The plasma of neuroborreliosis patients appeared to have an increased relative abundance of sphingomyelin CerPCho d18:1/24:1 and a decrease in CerPCho d18:0/18:0. Principal components analysis of the relative abundances of all PL class species distinguished between neuroborreliosis patients and healthy subjects. This is the first report comparing PL classes and their molecular species in neuroborreliosis patients and healthy subjects.

Immunoexpression of intermediate filaments and morphological changes in the liver and bile duct of rats infected with Fasciola hepatica.

We investigated the immunoexpression of the intermediate filament proteins, cytokeratin and desmin, and the morphological changes in the liver of rats during experimental fasciolosis at 4, 7 and 10 weeks post-infection. Rats were infected with 30 Fasciola hepatica metacercariae. Paraffin sections of the liver were stained using H & E, PAS and azan stains. Immunohistochemical reactions were performed using antibodies against cytokeratin and desmin. The experimental F. hepatica infection led to fibrosis and cirrhosis of the liver, and to inflammation of the common bile ducts. The expression of cytokeratin was increased in the epithelial cells of both the liver bile ductules at 4, 7 and 10 weeks post-infection and in the common bile ducts at 7 and 10 weeks post-infection compared to uninfected rats; expression in the common bile ducts was more intense. The myofibroblasts of the liver and smooth myocytes of the interlobular bile ducts and common bile ducts, showed a slight increase in desmin expression compared to the uninfected rats. The increased expression of cytokeratins in the hyperplastic rat common bile duct epithelium during the biliary phase of fasciolosis at 7 and 10 weeks post-infection may be explained by mechanical irritation by the parasite and an inflammatory reaction in the bile duct epithelium and in periductal fibrous tissue.

The redox status of human breast cancer cell lines (MCF-7 and MDA-MB231) treated with novel dinuclear berenil-platinum(II) complexes.

This study compared the effects of cisplatinum and novel berenil-platinum(ll) complexes on the redox status of breast cancer cells that were estrogen receptor-positive (MCF-7) or estrogen receptor-negative (MDA-MB231). Both cell lines were treated with cisplatinum or the following berenil-platinum(ll) complexes: Pt2(isopropylamine)4(berenil)2, Pt2(piperidine)4(berenil)2, Pt2(2-picoline)4(berenil)2, Pt2(3-picoline)4(berenil)2, and Pt2(4-picoline)4(berenil)2. Changes in levels of reactive oxygen species, levels and activities of antioxidants, and lipid peroxidation products levels were measured. All investigated compounds enhanced ROS generation, reduced the activity of antioxidant enzymes (e.g., glutathione peroxidase and glutathione reductase), and decreased levels of small-molecule antioxidants (GSH, vitamins E and A). Such conditions are conducive to generating oxidative stress and phospholipids peroxidation. Cellular phospholipids in MCF-7 cells were most sensitive to the Pt2(isopropylamine)4(berenil)2 complex, whereas MDA-MB231 cells were not particularly sensitive to any berenil-platinum(ll) complex. These findings will facilitate future anticancer drug design strategy for breast cancer pharmacotherapy.

Effects of dinuclear berenil-platinum(II) complexes on fibroblasts redox status.

PURPOSE: Platinum(II) complex anticarcinogenic mechanisms are associated with changes in the cellular redox status of cancer as well as healthy cells. Therefore, the goal of the present study was to investigate oxidative modifications in cellular components following fibroblast exposure to novel dinuclear berenil-platinum(II) complexes. MATERIAL AND METHOD: ROS levels, antioxidant parameters level/activity, and damage to DNA, lipids, and proteins, including pro-apoptotic and anti-apoptotic factors in human skin fibroblasts following berenil-platinum(II) complex treatments i.e. Pt2(isopropylamine)4(berenil)2, Pt2(piperazine)4(berenil)4, Pt2(2-picoline)4(berenil)2, Pt2(3-picoline)4(berenil)2, and Pt2(4- picoline)4(berenil)2 were examined. RESULTS: Treatment of fibroblasts with platinum(II) complexes has shown that all compounds enhance total ROS and superoxide anion generation as well as change the activity of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase and decrease in the level of non-enzymatic antioxidants (GSH, vitamin C, E and A). Such a situation is conducive to oxidative stress formation and oxidative modifications of cellular macromolecules and to increase in the expression of proapoptotic proteins. Pt2(isopropylamine)4(berenil)2 elicited the most damage, which resulted in oxidative modification of cellular components. The therapeutic use of this complex would cause considerable side effects in patients, therefore the agent lacks drug potential; however Pt2(piperazine)4(berenil)2 and Pt2(2-picoline)4(berenil)2 exhibited reduced redox and increased apoptotic profiles compared to cisplatin. CONCLUSION: Results of this paper and preliminary data show that Pt2(2-picoline)4(berenil)2 is less dangers than cisplatin to fibroblasts and more disruptive than cisplatin to breast cancer cell metabolism, and therefore it is a promising candidate for use in future anticancer drug strategies.

Effect of sweet grass extract against oxidative stress in rat liver and serum.

Ethanol metabolism is accompanied by generation of free radicals which can damage the cell components. However, sweet grass is a source of coumarin and its derivatives have emerged as a promising group of antioxidant compounds. The aim of this study has been to investigate the influence of sweet grass on oxidative stress formation in the liver and serum of rats intoxicated with ethanol. Alcohol intoxication led to a decrease in the superoxide dismutase, catalase, glutathione peroxidase and reductase activity in the blood serum as well as in the liver, but not in the glutathione reductase activity. The decrease in the antioxidant abilities of the examined tissues after ethanol intoxication resulted in enhanced lipid peroxidation measured as malondialdehyde and 4-hydroxynonenal levels. The metabolic consequence of oxidative modifications of lipids was damage of the liver cells membrane and an increase in its permeability appeared as a leakage of alanine aminotransferase and aspartate aminotransferase into the blood. Administration of sweet grass to the ethanol-intoxicated rats remarkably prevented the significant increase in concentrations of all measured lipid peroxidation products as well as the damage of the liver cell membrane. These results indicate beneficial antioxidant effect of the sweet grass on the liver of rats intoxicated with ethanol.

Age-dependent changes in the proteolytic-antiproteolytic balance after alcohol and black tea consumption.

Aging is accompanied by changes in the redox balance that is additionally modified by alcohol. Ethanol metabolism is connected with generation of free radicals which can damage cell components especially when antioxidant mechanisms are not able to neutralize them. In connection with the necessity of prevention against oxidative consequences, natural antioxidants are looked for. A natural and commonly used component of the diets with antioxidant properties are teas, especially the black tea. This study provides evidence of the role of black tea in the protection of rat plasma proteins and lipids against oxidative stress caused by aging and ethanol intoxication. For 5 weeks, the rats (2-, 12-, and 24-months old) used for the experiment received a black tea beverage (3 g/l) without or with alcohol (given for 4 weeks). The decrease in antioxidant abilities determined as total antioxidant status during aging and ethanol intoxication resulted in enhanced lipid and protein oxidation (determined as malondialdehyde, carbonyl groups, dityrosine, tryptophan and sulfhydryl groups level). In consequence the decrease in anti-proteases (alpha-1-antitrypsin, alpha-2-macroglobulin) activity and the increase in proteases (elastase and cathepsin G) activity were observed. Black tea protected the plasma antioxidants and prevented oxidative modifications of lipid and protein observed during aging as well as ethanol intoxication. The results indicate that a shift into plasma proteolytic activity results from a decrease in antioxidant abilities, so the use of black tea appears to be beneficial in reducing oxidative stress caused by ethanol and/or aging.

Lipid peroxidation products as potential bioindicators of Lyme arthritis.

Lipid peroxidation products, malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and [Formula: see text], were determined in the plasma and urine of patients with Lyme arthritis and healthy people. The group consisted of 19 patients with Lyme arthritis (mean age 47 years) and the control group consisted of 16 healthy individuals (mean age 38 years). Diagnosis of Lyme disease was confirmed by epidemiological anamnesis, clinical manifestation of arthritis and serological examinations. Lipid peroxidation was estimated by the measurement of aldehydes (MDA and 4-HNE, determined by high-performance liquid chromatography [HPLC]) and prostaglandin derivatives (8 - isoPGF(2a), determined by liquid chromatography/mass spectrometry [LC/MS]). MDA and 4-HNE levels were increased about 2-4-fold in the plasma, while in the urine, the increases were about 2-fold. More significant increases were noted for the 8 - isoPGF(2a) total plasma level, which was enhanced over 4-fold, and for the urine 8 - isoPGF(2a) level, which was increased over 8-fold. The 8 - isoPGF(2a) total plasma level consists of free and esterified form. During infection, the ratio of free to esterified form is significantly smaller compared to healthy people. The ratio of free to esterified form of 8 - isoPGF(2a) may be a useful indicator of Lyme arthritis. Moreover, the complementarities of three lipid peroxidation product levels may be helpful in the diagnosis of Lyme arthritis.

Capillary zone electrophoresis method for determination of bitter (alpha- and beta-) acids in hop (Humulus lupulus L.) cone extracts.

PURPOSE: Humulus lupulus (H. lupulus), more commonly known as hop, is a member of the Cannabaceae family with male and female flowers on separate plants. It is native in Europe including Lithuania, Asia and North America. Hop has been recognized as a medicinal plant for centuries, nevertheless different medicinal activities of hop are currently investigated and discovered. An important class of hop compounds is the hop acids, which are classified as alpha-acids and beta-acids. Different varieties of hops vary in amount and composition of hop acids. METHODS: Simple capillary zone electrophoresis method has been optimized and applied for the analysis of hop acids in hop cone extracts. RESULTS: With this method the analysis takes ca. 10 min. Repeatability for migration times and peak areas expressed as relative standard deviation were up to 0.21% and 5.96%, respectively. CONCLUSIONS: Comparative results of capillary zone electrophoretic analysis of extracts of different hop varieties and conductometric titration, as a standard method for determination of alpha-acids, are presented. Both methods provide consistent results, however capillary zone electrophoresis is capable of separating co- form of humulones from other forms.

Oxidative stress and antioxidative defense parameters early after reperfusion therapy for acute myocardial infarction.

Reperfusion of ischemic myocardium evokes rapid release of free radicals in experimental models. The aim of the study was to investigate the oxidative stress and antioxidative defense during first minutes after reopening of the infarct related artery in patients treated for acute myocardial infarction. The study group consisted of 15 patients with first ST elevation myocardial infarction (STEMI) due to left anterior descending artery occlusion. The control group included ten patients with stable ischemic heart disease (IHD). Blood samples from coronary sinus were drawn before, immediately after and about 15 min after angioplasty. Activity of superoxide dysmutase (SOD), concentration of glutathione as well as the concentrations of lipid peroxides, malodialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE) were measured. There was significantly higher concentration of MDA and HNE and higher SOD activity in STEMI patients before the reperfusion, as compared to the stable IHD group. After the reperfusion concentration of HNE in erythrocytes from STEMI patients was higher than in IHD group. At the same time the activity of SOD significantly decreased in patients with impaired tissue perfusion (myocardial blush grade <2). In conclusion, there is a slightly higher concentration of oxidative stress parameters in patients with STEMI. Diminished antioxidative defense after reperfusion is associated with impaired myocardial perfusion.

The comparison of effect of catechins and green tea extract on oxidative modification of LDL in vitro.

PURPOSE: Green tea due to its content of catechins reveals strong antioxidative activity, which is manifested among others by its ability to inhibit free radical generation, scavenge free radicals as well as chelate transition metal ions that catalyse free radical reactions. The influence of green tea extract, epicatechin (EC), epicatechin galate (ECG) as well as epigallocatechin galate (EGCG) on oxidative modifications of LDL of human blood serum has been examined in the present study. MATERIALS AND METHODS: This influence has been evaluated by measurement of the concentration of first products of lipid peroxidation--conjugated dienes and lipid hydroperoxides as well as by determining tryptophan and dityrosine content-- the markers of protein oxidative modification. RESULTS: Catechins and green tea abilities to protect lipophilic antioxidant--alpha-tocopherol against oxidation have been also examined. The results reveal that peroxidation of LDL is markedly prevented by green tea extract and in a slightly weaker way by catechins (EGCG in particular), which is manifested by a decrease in concentration of conjugated dienes, lipid hydroperoxides, MDA, dityrosine and by an increase in tryptophan content. Both green tea as well as catechins (EGCG in particular) have been also revealed to prevent decrease in concentration of alpha-tocopherol in oxidating conditions. CONCLUSIONS: It can be assumed that green tea and to a lesser degree catechins, protecting the basic antioxidant of LDL-alpha-tocopherol, prevent oxidative modification of LDL.

Antioxidant potential of rat liver in experimental infection with Fasciola hepatica.

The aim of this paper is to assess the antioxidant properties of rat liver in the course of acute and chronic fasciolosis. Wistar rats were infected per os with 30 metacercariae of Fasciola hepatica. Liver activities of antioxidant enzymes and concentrations of non-enzymatic antioxidants were determined at 4, 7, and 10 weeks post-infection. Activities of superoxide dismutase (Cu,Zn-SOD), glutathione peroxidase (GSH-Px), and glutathione reductase (GSSG-R) were decreased, catalase (CAT) activity was increased and non-enzymatic antioxidant concentrations (reduced glutathione, vitamins C, E and A) were reduced simultaneously with enhancement of lipid peroxidation processes as evidenced by increased levels of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). Changes in the antioxidant abilities of the liver and in the phospholipid structure of the cell membrane were accompanied by rising activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as markers of liver damage.

Influence of green tea on surface charge density and phospholipids composition of erythrocytes membrane in ethanol intoxicated rats.

Ethanol is oxidized to acetaldehyde and then to acetic acid and these processes are accompanied by free radical generation. This paper reports the effect of green tea on electric charge and phospholipids composition of erythrocytes membrane from rats intoxicated with ethanol. Electrophoresis technique and HPLC have been applied to above-mentioned studies. Ethanol administration caused increase in erythrocyte membrane surface charge density and phospholipid composition. Ingestion of green tea with ethanol partially prevented changes in structure and function of membrane caused by chronic ethanol intoxication.

Antioxidative properties of black tea.

BACKGROUND: Black tea, obtained by tea leaves fermentation, is an oxidized product and contains mainly multimeric polyphenols, whose biological activity is not well documented. This paper reviews the available literature on the effects of black tea on health with a focus on its antioxidative activity. METHODS: A review of the different issues and studies relating to composition, manufacturing, and antioxidative effects of black tea and its components in vitro as well as in vivo is presented. RESULTS: It is generally believed that polyphenols such as theaflavins and thearubigins as well as catechins as major constituents of black tea are mainly responsible for antioxidant actions. Antioxidative properties of black tea are manifested by its ability to inhibit free radical generation, scavenge free radicals, and chelate transition metal ions. Black tea, as well as individual theaflavins, can influence activation of transcription factors such as NFkappaB or AP-1. Theaflavins have been also proved to inhibit the activity of prooxidative enzymes such as xanthine oxidase or nitric oxide synthase. CONCLUSIONS: Black tea consumed throughout the world is believed to be not only a popular beverage but also an antioxidative agent available in everyday life.

Antioxidant properties of black tea in alcohol intoxication.

Food ingredients such as alcohol may modify cellular redox state. Ethanol metabolism is accompanied by generation of free radicals that can damage cell components especially when antioxidant mechanisms are no able to neutralize them. However black tea is a source of polyphenol antioxidants that may enhance cellular antioxidant abilities. The aim of this study was to investigate the effect of black tea on antioxidant abilities of the liver, blood serum and brain of 12-months old rats sub-chronically (for 28 days) intoxicated with ethanol. Administration of black tea alone caused increase in the activity and concentration of antioxidant parameters more extensively in the liver and serum than in the brain. Alcohol caused decrease in the liver glutathione peroxidase and reductase and catalase activity but increase in activity of superoxide dismutase. Moreover, decrease in the level of non-enzymatic antioxidants, such as reduced glutathione, vitamin C, A and E and beta-carotene was observed. The activity of serum glutathione peroxidase and reductase decreased while superoxide dismutase activity was not changed. The level of non-enzymatic antioxidants in serum was also decreased. However brain activity/level of all examined antioxidants enzymatic as well as non-enzymatic was decreased after ethanol intoxication. Black tea considerably prevented antioxidant parameters against changes caused by ethanol. These results indicate beneficial antioxidant effect of black tea regarding all examined tissues, but especially the liver.

Protein Z and vitamin K in kidney disease.

PURPOSE: Disturbances in hemostasis are common complications of kidney diseases. Both bleeding diathesis and thromboembolism may complicate the course of chronic uremia. As far as we know, there is a limited data about protein Z in kidney disease. MATERIAL AND METHODS: The aim of our work was to examine plasma protein Z and vitamin K concentrations in nephrotic syndrome (n = 34), glomerulonephritis (n = 48), kidney transplant recipients (n = 80), peritoneally dialyzed patients (n = 42) and in the healthy volunteers (n = 27). RESULTS: Vitamin K was significantly lower in nephrotic syndrome when compared to non-nephrotic patients, CAPD and healthy volunteers (p < 0.05). Protein Z was the highest in CAPD and kidney transplant recipients when compared to any other group. In nephrotic syndrome protein Z was significantly lower when compared to the healthy volunteers, but it did not differ significantly between two groups of patients with chronic renal failure (with and without nephrotic syndrome). Protein Z correlated only with fibrinogen in CAPD, glomerulonephritis and nephrotic patients. Vitamin K correlated with age and albumin in patients with glomerulonephritis, nephrotic syndrome as well as with albumin in CAPD. CONCLUSIONS: Alterations in protein Z might contribute to the enhanced risk of thromboembolic complications in nephrotic syndrome, CAPD and Tx via different and unknown mechanisms. This phenomenon seems to be unrelated to vitamin K status in these patients.

Release of hydroxyl ions from calcium hydroxide preparations used in endodontic treatment.

PURPOSE: The purpose of our study was to compare the in vitro release of hydroxyl ions from several calcium hydroxide preparations used in endodontic treatment. MATERIAL AND METHODS: Equal quantities of the materials--nonsetting (pure calcium hydroxide, Biopulp, Calcicure), setting--canal sealers (Sealapex, Apexit) and points were placed in dialysis tubes which were then immersed in deionized water. The release of hydroxyl ions from the preparations was measured by the median pH of the deionized water used for dialysis, by means of a pH-meter. The results of our study were analyzed by means of Tukey's reasonable correlation. Significance difference (one-way variance analysis ANOVA) and Pearson's linear correlation coefficient (r2). RESULTS: Nonsetting preparations of calcium hydroxide have a significantly higher capability of hydroxyl ions release in comparison with sealers and points, irrespective of time (p < 0.05). Sealapex and "plus" points released hydroxyl ions to a much greater extent than both Apexit and "regular" points at most periods of the experiment (p < 0.05). Apexit released significantly more of hydroxyl ions than "regular" points, and Sealapex more than "plus" points in the later periods of the experiment (p < 0.05). The pH values of dialysis samples of all materials correlated positively with time and the pH. Almost all materials reached a maximum on the 8-th day of the experiment. CONCLUSIONS: To achieve maximum concentration of hydroxyl ions in tissues: for temporary root fillings nonsetting preparations of calcium hydroxide should be chosen rather than points and they should be placed for at least one week, for permanent root fillings it is more recommended to use Sealapex than Apexit as a sealer.

Green tea as a potent antioxidant in alcohol intoxication.

Ethanol oxidation to acetaldehyde and next to acetate is accompanied by free radical generation. Free radicals can affect cell integrity when antioxidant mechanisms are no longer able to cope with the free radical generation observed in ethanol intoxication. Natural antioxidants are particularly useful in such a situation. The present study was designed to investigate the efficacy of green tea as a source of water-soluble antioxidants (catechins) on the liver and blood serum antioxidative potential of rats chronically (28 days) intoxicated with ethanol. Alcohol caused a decrease in liver superoxide dismutase, glutathione peroxidase and catalase activities and an increase in activity of glutathione reductase. Moreover, a decrease in the level of reduced glutathione, ascorbic acid, vitamins A and E and beta-carotene were observed. The activity of serum glutathione peroxidase decreased while glutathione reductase activity increased. The level of serum non-enzymatic antioxidants was also decreased in the liver. Alcohol administration caused an increase in the liver and serum lipid peroxidation products, measured as thiobarbituric acid-reactive substances. However, green tea prevents the changes observed after ethanol intoxication. Green tea also protects membrane phospholipids from enhanced peroxidation. These results indicate a beneficial effect of green tea in alcohol intoxication.

Protective effect of green tea against lipid peroxidation in the rat liver, blood serum and the brain.

This paper reports data on the effect of green tea on the lipid peroxidation products formation and parameters of antioxidative system of the liver, blood serum and central nervous tissue of healthy young rats drinking green tea for five weeks. The rats were permitted free access to solubilized extract of green tea. Bioactive ingredients of green tea extract caused in the liver an increase in the activity of glutathione peroxidase and glutathione reductase and in the content of reduced glutathione as well as marked decrease in lipid hydroperoxides (LOOH), 4-hydroksynonenal (4-HNE) and malondialdehyde (MDA). The concentration of vitamin A increased by about 40%. Minor changes in the measured parameters were observed in the blood serum. GSH content increased slightly, whereas the index of the total antioxidant status increased significantly. In contrast, the lipid peroxidation products, particularly MDA was significantly diminished. In the central nervous tissue the activity of superoxide dismutase and glutathione peroxidase decreased while the activity od glutathione reductase and catalase increased after drinking green tea. Moreover the level of LOOH, 4-HNE and MDA significantly decreased. The use of green tea extract appeared to be beneficial to rats in reducing lipid peroxidation products. These results support and substantiate traditional consumption of green tea as protection against lipid peroxidation in the liver, blood serum, and central nervous tissue.