Effects of antihistamines on isolated human lung mast cells, basophil leucocytes and erythrocytes

Low concentrations of H1- and H2-receptor directed compounds inhibited the immunologic secretion of histamine from human lung mast cells and basophil leucocytes and protected human erythrocytes from hypotonic lysis. Higher concentrations induced a release of the amine from the histaminocytes. These results are discussed in terms of the possible membrane effects of the test compounds.     

Direct and metabolism-dependent toxicity of sulphasalazine and its principal metabolites towards human erythrocytes and leucocytes.

1. The role of metabolites in sulphasalazine-mediated toxicity has been investigated in vitro by the use of human red blood cells and mononuclear leucocytes as target cells, with methaemoglobin formation and cytotoxicity respectively, being the defined toxic end-points. 2. Of the metabolites of sulphasalazine investigated, only sulphapyridine was bioactivated by human liver microsomes in the presence of NADPH to a metabolite which caused marked methaemoglobinaemia and a small, but statistically significant degree of mononuclear leucocyte cell death. 3. Methaemoglobinaemia was inhibited by ketoconazole but not by ascorbic acid (100 microM), glutathione (500 microM) and N-acetylcysteine (50 microM). In contrast, ascorbic acid and the thiols afforded complete protection for mononuclear leucocytes. 4. Sulphapyridine (100 microM) was converted in vitro to a metabolite (metabolite conversion 6.8 +/- 0.3%), the retention time of which on h.p.l.c. corresponded to synthetic sulphapyridine hydroxylamine. The half-life of sulphapyridine hydroxylamine in phosphate buffer (pH 7.4) was found to be 8.1 min. 5. In the absence of microsomes and NADPH, sulphapyridine hydroxylamine caused a concentration-dependent (10-500 microM) increase in methaemoglobinaemia (2.9%-24.4%) and cytotoxicity (5.4%-51.4%), whereas sulphasalazine, sulphapyridine, 5-hydroxy sulphapyridine and 5-aminosalicylic acid had no effect.     

Vitamin E protects human leucocytes against toxic effects of lindan in vitro.

Vitamin E (15 micrograms/ml and 60 micrograms/ml) was used for protection of human leucocytes against toxic effects of lindan (150 microM and 200 microM) in vitro. The protective effect of vitamin E manifested itself as maintenance of the phagocytic activity of granulocytes, the value of nucleologram in lymphocytes, and the proportion of euchrysin-negative lymphocytes near the values in controls.     

The toxic effect of opioid analgesics on human sperm motility in vitro

Opioid analgesics are the most common therapeutic analgesic for acute pain. In this study, the toxicological and pharmacological features of a group of opioid analgesics were characterized by the motility of human sperm. Aliquots of sperm were incubated with various concentrations of opioid analgesics in vitro. Computer-assisted sperm analysis was used to assess sperm motility at 15 minutes, 2 hours, and 4 hours after drug addition to the medium. Butorphanol and dezocine showed marked reduction of motility after incubation with sperm for 15 minutes. Butorphanol was more effective than dezocine in immobilizing sperm. Other opioids studied, such as fentanyl, alfentanil, and sufentanil, showed only partial inhibitory activity. Based on the data reported herein, we have found that butorphanol and dezocine exert a sperm-immobilizing effect. However, fentanyl, alfentanil, and sufentanil exhibit only partial inhibition of sperm motility. Given the increasing use of opioids and their potential effect on sperm motility, these findings are greatly relevant to male reproductive health.     

The effect of acute ethanol ingestion on in vitro metabolism of choline and ethanol derivatives in rat liver

The effect of acute ethanol ingestion on hepatic phospholipid metabolism has been investigated in the rat. No significant increase in the content of either lecithin or phosphatidylethanolamine occured in the liver after 12 hr from treatment. The triglyceride content of the liver increased three-fold. An increase of lecithin and phosphatidylethanolamine synthesis by the Kennedy pathway was found in vitro in the homogenates and microsomal fractions prepared from the ethanol-treated rats. A higher rate of conversion of phosphorylcholine and phosphorylethanolamine to CDP-choline and CDP-ethanolamine was also found in the experimental animals. The breakdown of CDP-choline to phosphorylcholine was noticeably decreased in the liver fractions of the ethanol-treated rats. No changes in the sequential methylation pathway for lecithin synthesis in the liver were observed after acute ethanol ingestion.     

The effect of thyroid hormones and other kinetic modifiers on bisphosphoglyceromutase from human erythrocytes

Bisphosphoglyceromutase (E.C. 2.7.5.4) is inhibited by thyroxine, tri-iodothyronine and diiodothyronine and activated by glucagon. Several other hormones have no effect. The enzyme is activated by various thiols, but not by NADH, NADPH or ascorbic acid. The inhibition by α- and β-glycerophosphate and the activation by phosphoglycollic acid [1] have been confirmed. 2-Hydroxy-4-phosphobutyric acid, an isostere of β-glycerophosphate, has no effect on the kinetics. Nicotine is slightly inhibitory at concentrations much higher than the plasma level in smokers.     

The effect of ifosfamide and its metabolites on intracellular glutathione levels in vitro and in vivo

The effect of ifosfamide and its metabolites on intracellular levels of glutathione in P388 cells in vitro has been studied. It is demonstrated that glutathione depletion occurs only in the presence of 4-hydroperoxyifosfamide and chloroacetaldehyde. In contrast isophosphoramide mustard had no effect on glutathione levels in intact cells. The concentration of 4-hydroperoxyifosfamide required to reduce glutathione levels by 50% was approximately 1 mM and this represents a concentration far in excess of that achievable in patients receiving the drug. However the concentration of chloroacetaldehyde (approximately 100 microM) required to reduce intracellular levels of glutathione to a similar extent is attained in patients receiving ifosfamide. The glutathione levels in lymphocytes isolated from a patient undergoing an eight hour infusion of ifosfamide showed a marked decrease to about 30% of their original value. We conclude that ifosfamide causes glutathione depletion in vivo and the majority of this can be accounted for by the production of chloroacetaldehyde.     

Effects of nicotine,its metabolites and tobacco extracts on human platelet function in vitro

Cigarette smoking is a leading cause of cardiovascular disease. The cardiovascular effects of smoking are probably multifactorial, including effects on platelets. Previous reports investigating the effects of nicotine and tobacco on platelet function are inconsistent.The present study investigated in vitro effects of nicotine, its major metabolites, tobacco extracts and extract of tobacco-free snuff on human platelets.None of the metabolites cotinine, cotinine-N-oxide, nicotine-1′-N-oxide or trans-3′-hydroxycotinine (0.1–10 μM) affected platelet aggregation or P-selectin expression. Nicotine (10 μM) weakly increased platelet aggregation, whereas trans-3′-hydroxycotinine (0.1 μM) and nicotine-1′-N-oxide (1–10 μM) weakly inhibited adhesion to fibrinogen. To elucidate the influence of other tobacco compounds, we investigated the impact of moist tobacco and smoke extracts on platelet function. Filtered extracts of oral snuff, cigarette smoke and tobacco free snuff inhibited platelet adhesion concentration-dependently. The inhibitory effects of tobacco extracts on platelet adhesion were independent of nicotine content and the nitric-oxide-pathway and not mediated through a platelet-nicotine-receptor.Taken together, tobacco extracts inhibit platelet activation during short-term in vitro challenge. As only limited effects of nicotine and nicotine metabolites were seen, the tobacco-induced platelet inhibition are likely induced by other compounds present in tobacco and tobacco free snuff.     

Spin trapping of free radical metabolites of carbon tetrachloride in vitro and in vivo: effect of acute ethanol administration.

A single dose of ethanol, when administered 18 hr prior to CCl4, potentiates the hepatotoxicity of the halocarbon. In these studies, spin trapping and electron spin resonance (ESR) spectroscopy methods were utilized to determine whether a single ethanol dose increased the metabolism of CCl4 to free radical intermediates. When hepatic microsomes from ethanol-treated or control rats were incubated with CCl4 and the spin trapping agent alpha-phenyl-N-tert-butylnitrone (PBN), the ESR signal of the trichloromethyl radical adduct of PBN was of similar intensity in both groups. The ethanol dose also failed to induce p-nitrophenol hydroxylase activity. When PBN and CCl4 were administered to rats, liver extracts contained ESR signals resulting primarily from the trichloromethyl radical adduct of PBN, and the signals were of similar intensity in both experimental groups. Higher concentrations of the carbon dioxide anion radical adduct of PBN were detected in plasma samples from ethanol-treated rats. However, when hepatocytes from ethanol-treated and control rats were incubated with PBN and CCl4, ESR signals of the carbon dioxide adduct were of similar intensity. These data suggest that the higher concentrations of the carbon dioxide adduct in the blood of ethanol-treated rats may be explained by early CCl4-induced damage to liver cell membranes, rather than increased rates of formation. The data in this report fail to support the hypothesis that a single dose of ethanol stimulates the hepatic metabolism of CCl4 to the trichloromethyl radical. Alternatively, ethanol may potentiate CCl4 toxicity by affecting some critical metabolic step subsequent to trichloromethyl radical formation.     

In vitro effect of xipamide on sodium-potassium transport systems in human erythrocytes

The in vitro effects of xipamide in a concentration range of 10(-8) to 10(-2) M were investigated on various Na+ and K+ transport systems in human red blood cells. Xipamide inhibited the anion carrier or DIDS-sensitive LiCO3- -influx starting from a concentration of 10(-5) M. However, a decrease in the Na+, K+-pump and the Na+, K+-cotransport activity and a rise in the passive permeability of the cell membrane was only observed starting from a concentration of 10(-4) M xipamide.     

In vitro effect of cilazaprilat on sodium-potassium transport systems in human erythrocytes

The in vitro effects of cilazaprilat in a concentration range of 10 nM to 1 mM were investigated on various Na+ and K+ transport systems in human red blood cells. Cilazaprilat inhibited the anion carrier or DIDS-sensitive LiCO3(-)-influx and the bumetanide-sensitive K(+)-efflux. However, no effect of cilazaprilat on the Na+, K(+)-pump activity, the number of active Na+ pump units and on the Na+, Li(+)-countertransport activity could be demonstrated. It is suggested that through a decreased anion carrier indicating a decreased Na(+)-influx, the lower intracellular Na+ concentration observed in vivo during angiotensin-converting enzyme inhibition, can be at least partly explained.     

The effect of secretin on sodium ion absorption by the isolated human gallbladder

Sodium ion (Na+) transport, a principal function of the gallbladder epithelium, was studied by measuring the flux of 22Na across isolated human gallbladder mucosa maintained in a modified 'Ussing' flux chamber. Tissue was obtained from cholecystectomy specimens in symptomatic patients with cholelithiasis. Out of 26 gallbladders studied, 13 had a net Na+ flux from mucosa to serosa which indicated active Na+ absorption. The hormone secretin, when added to the serosal fluid, reversed the direction of net flux in these gallbladders and caused a secretion of Na+ from serosa to mucosa. These results suggest that secretin may be involved in the physiological regulation of fluid transport in the human gallbladder, and also suggest a possible role for this hormone in gallbladder emptying.     

The toxic effect of thioacetamide on rat liver in vitro

Thioacetamide (TAA) is a hepatotoxin frequently used for experimental purposes which produces centrilobular necrosis after a single dose administration. In spite of the fact that oxidative stress seems to play a very important role in the mechanism of TAA-induced injury, the effect of TAA on hepatocytes in primary culture with respect to the influence on mitochondria has yet to be verified.Hepatocytes were incubated for 24 h in a medium containing TAA (0–70 mmol/l). Glutathione content (GSH/GSSG), reactive oxygen species and malondialdehyde formation were assessed as markers of cell redox state. Toxicity was determined by lactate dehydrogenase leakage and WST-1 assay. The functional capacity of hepatocytes was evaluated from albumin and urea production. Mitochondrial metabolism was assessed by measuring mitochondrial membrane potential and oxygen consumption.Our results show that a profound decrease in the GSH level in hepatocytes precedes a sharp rise in endogenous ROS production. ROS production correlates with an increase in lipoperoxidation. Mitochondria are affected by TAA secondarily as a consequence of oxidative stress. Oxidation of the NADH-dependent substrates of respiratory Complex I is significantly more sensitive to the toxic action of TAA than oxidation of the flavoprotein-dependent substrate of Complex II. Mitochondria can also maintain their membrane potential better when they utilize succinate as a respiratory substrate. It appears that GSH should be depleted below a certain critical level in order to cause a marked increase in lipid peroxidation. Mitochondrial injury can then occur and cell death develops.     

The effect of alendronate sodium on human erythrocytes

Alendronate sodium is a medicine, which is commonly used in osteoporosis treatment. Nowadays, this substance is given to patients in tablets, but in future it is planned that it will be administrated into human organisms as intravenous infusions; therefore, significant interactions of this medicine with erythrocytes will be inevitable. It is the reason why we decided to investigate the interaction of alendronate sodium with human erythrocytes. The effect of this medicine on acetylcholinesterase activity, lipid and protein peroxidation, as well as cellular thiol content was examined. Moreover, the effect of alendronate sodium on alterations in erythrocytes morphology was assessed. Human erythrocytes were incubated with alendronate sodium in the concentrations ranging from 0.33 to 100 μM for 1 h and 24 h. No changes have been observed in the parameters examined after 1 h of incubation of the erythrocytes with this medicine excluding the carbonyl groups level. Moreover, no alterations in the activity of acetylcholinesterase, the level of thiols as well as in morphology of the erythrocytes incubated with alendronate sodium for 24 h have been observed. It was also proven that alendronate sodium increased the level of reactive oxygen species (ROS) but only after 24 h of incubation. We have not observed any severe changes in cells studied even at the highest dose of bisphosphonates examined, thus their usage should not be dangerous for the erythrocytes of people treated with these medicines.     

The toxic and metabolic effects of 23 aliphatic alcohols in the isolated perfused rat liver.

We investigated the acute toxic and metabolic effects of 23-aliphatic alcohols (16 saturated and 7 unsaturated) in the isolated perfused rat liver at a concentration of 65.1 mmol/l (approximately 0.3% ethanol). The capacity of the straight chain primary alcohols (methanol, ethanol, 1-propanol, 1-butanol and 1-pentanol) to release the enzymes glutamate-pyruvate transaminase (GPT), lactate dehydrogenase (LDH) and glutamate dehydrogenase (GLDH) into the perfusate was strongly correlated with their carbon chain length. The secondary alcohols were less active in this respect whereas branching of the carbon chain did not consistently change alcohol toxicity. Unsaturation in the straight chain but not in the branched chain alcohols was accompanied by an increase in toxicity. An increased enzyme release was in general accompanied by, and correlated to, reductions in oxygen consumption, bile secretion, and perfusion flow of the isolated livers. Statistically significant correlations exist between parameters of alcohol-induced hepatotoxicity and the membrane/buffer partition coefficents of the alcohols. With the exception of methanol, all alcohols tested increased the lactate/pyruvate ratio of the perfusate, although this effect was not correlated to the degree of hepatic injury. Hepatic ATP concentrations decreased in most cases in line with hepatic injury and were particularly correlated with changes in oxygen consumption. Hepatic concentrations of reduced glutathione (GSH) were only diminished by the unsaturated alcohols, whereas an increase in hepatic oxidized glutathione (GSSG) occurred only with some of the saturated alcohols. Hepatic concentrations of malondialdehyde (MDA) increased after two saturated and three unsaturated alcohols but did not correlate with other parameters of hepatotoxicity. In conclusion, alcohol-induced hepatotoxicity is primarily due to membrane damage induced by the direct solvent properties of the alcohols. The consequences and relative contributions of alcohol metabolization to the overall hepatotoxicity of higher alcohols requires further study.     

The acute effect of ethanol on behaviour,body temperature,and brain histamine in mice

The possible relationships between ethanol-induced changes in behaviour, body temperature, and brain histamine were studied. Mice were injected intraperitoneally with various doses of ethanol in the range of 0.088–1.75 g/kg. There was a dose-dependent biphasic alteration in behaviour and body temperature, where a low dose of 0.175 g/kg produced excitation and hyperthermia. Doses of 0.875 and 1.75 g/kg caused an increasing state of depression and dose-dependent hypothermia. Behavioural changes induced by ethanol appear to reflect changes in body temperature. When hyperthermia was produced the animals were hyperactive, whereas with hypothermia they were sedated and huddled close together. These changes in behaviour and body temperature closely paralleled the ethanolinduced modifications in whole brain histamine levels, indicating that a relationship may exist between the ethanol-induced changes in these parameters.     

The effect of cannabidiol,alone and in combination with ethanol,on human performance

Fifteen volunteers received cannabidiol (CBD) (320 g/kg) or placebo (both orally, T₀), and 60 min later they consumed an ethanolic beverage (0.54 g/kg) or placebo. The effects were measured at T₁ (100 min after CBD ingestion), T₂ (160 min) and T₃ (220 min) using cognitive, perceptual and motor function tests. Factorial analysis indicated that test procedures could be adequately expressed by three rotated factors: A reaction speed factor (I), a standing steadiness factor (II) and a psychomotor coordination/cognitive factor (III). Ethanol produced a significant decrement in factor III. There was no demonstrable effect of CBD, either alone or in combination with ethanol. Neither CBD nor ethanol produced any significant effect on pulse rate. Prior administration of CBD did not significantly affect the blood ethanol levels. Whilst the subjects were able to identify correctly when they were given ethanol, they did not report any subjective effects of CBD.     

Protective effect of desloratadine against oxidative stress in human erythrocytes in vitro

Desloratadine (DCL) is a non-sedating antihistamine approved for the treatment of allergic rhinitis or chronic idiopathic urticaria. The objective of this study was to evaluate the potential protective effect of DCL against oxidative stress in human erythrocytes in vitro. Human erythrocytes were oxidized by a water-soluble radical generators-2,2' azobis (2-amidinopropane) hydrochloride (AAPH; 20, 50mM) or tert-butyl hydroperoxide (TBHP; 0.5mM) and the protective effects of DCL (2, 5, 7, 10 and 26μM) on selected oxidative stress markers were investigated. Erythrocytes were divided into aliquots. The first aliquot was incubated for 2h at 37°C with AAPH or TBHP. The other test aliquots were preincubated with selected concentrations of DCL for 30min and followed by AAPH or TBHP incubation for 2h. Malondialdehyde (MDA) content, catalase (CAT) and superoxide dismutase (SOD) activities, as well as hemolysis percentage (H) were measured in all erythrocyte samples. The influence of solvent (0.5% ethanol) on the parameters studied was also checked. Pretreatment with DCL (7, 10, 26μM) could prevent TBHP-induced increase in MDA formation in a concentration-dependent manner. DCL has no influence on CAT activity and it significantly enhanced SOD activity compared to AAPH treatment samples at 7, 10, 26μM. DCL (26μM) also reduced the hemolytic effect on erythrocytes when compared to the erythrocytes exposed to oxidants only. These results suggest a beneficial effect of DCL as an antioxidant, which might be an additional explanation of its therapeutic action.