Increase in non-transferrin bound iron and the oxidative stress status in epilepsy patients treated using valproic acid monotherapy.

OBJECTIVE: This study aims to investigate the alteration of iron homeostasis and oxidative stress status in epilepsy patients treated with valproic acid (VPA) monotherapy. MATERIALS: 24 epilepsy patients receiving VPA monotherapy (12 men, 12 women, age 27.5 ± 7.2 y) and 24 sex- and age-matched healthy volunteers were included in the study. METHODS: The level of iron status parameters; serum iron, ferritin, transferrin saturation, non-transferrin bound iron (NTBI), serum level of trace elements (copper, zinc and selenium), concentration of antioxidant parameters, activities of antioxidant enzymes and level of lipid peroxidation product were determined. RESULTS: NTBI was found in the patients although their other iron status parameters were normal. Levels of antioxidant parameters were decreased while activities of antioxidant enzymes were increased. Levels of serum MDA were significantly increased in patients with epilepsy. The daily dose of valproic acid associated was statistically significant: serum concentration of NTBI (r = 0.579; p = 0.003) and MDA (r = 0.465; p = 0.022). A positive correlation existed between NTBI and zinc (r = 0.522; p = 0.009). CONCLUSION: According to our results, VPA treatment in patients with epilepsy contributes to the metabolism of iron, leading to the formation of NTBI and an increase in oxidative stress.

Prevention of cerumen impaction by treatment of ear canal skin. A pilot randomized controlled study.

OBJECTIVES: To evaluate the effect of topical skin treatment of the ear canal in the prevention of impacted earwax. DESIGN: The study was prospective, randomized and controlled. SETTING: Secondary ORL care and Institute for the Mentally Retarded. PARTICIPANTS: Thirty-nine consecutive patients aged 1-74 years. Selection criteria were impacted earwax more than once a year and impacted earwax completely obstructing the lumen of the ear canal at the time of inclusion. Patients were randomized to active therapy or no intervention. The active therapy was Ceridal lipolotion instilled in the ear canal with a syringe once a week for 12 months. In the control group, earwax was removed without other intervention. The lipolotion was administered by the participants themselves with exception of the mentally retarded who were treated by nurses. Follow-up visits were at 3 and 12 months. MAIN OUTCOME: The main outcome was recurrence of impacted earwax with complete obstruction of the ear canal. There was significantly lower recurrence rate of impacted earwax in the treated compared with the control group (23%versus 61%, P < 0.05). CONCLUSIONS: This study suggests that prophylactic treatment of the ear canal with a topical emollient may prevent formation of impacted earwax.

Evaluation of the antibacterial and hemolytic activities of Latvian herbal preparation.

Three extracts originating from a combination of various Latvian plant species were tested for their antibacterial activities by evaluating growth delays using a fully automated microturbidimetric method. Ten different human and bovine strains of the genera Staphylococcus and Micrococcus were used as test microorganisms. The inhibitory effect in vitro was defined as the difference between the growth rate without herbs and the growth rate in the presence of an extract. Among the tested strains, Staphylococcus aureus was found sensitive to all 3 extracts. However, extract I was the most effective in slowing the growth of all strains tested. Using appropriate tester strains it should be possible to set up a broad-range microtubidimetry assay for individual herb screening in vitro. The hemolytic effects of the individual extracts on human erythrocytes were also studied at different concentrations. Two of the herbal extracts had minimal lytic effects on eurocaryotic cells. An additional hemolysis test was conducted in the presence of coenzyme Q10 (CoQ10) as a free radical scavenger: CoQ10 had no effect on the hemolytic reaction.

Measurement of antibacterial activities of T-2 toxin, deoxynivalenol, ochratoxin A, aflatoxin B1 and fumonisin B1 using microtitration tray-based turbidimetric techniques.

Various mycotoxins were tested for their antibacterial activity by evaluating growth delays using a fully automated microturbidmetric method. Ten different strains of the genera Escherichia, Streptococcus, Staphylococcus, Yersinia, Salmonella, Erysipelothrix and Lactobacillus were used as test micro-organisms. T-2 toxin, deoxynivalenol (DON), ochratoxin A (OTA), aflatoxin B1 (AFB1) and fumonisin B1 (FB1) were used as representative mycotoxins. The inhibitory effect in vitro was defined as the difference between the growth rate without mycotoxins and the growth rate in the presence of a mycotoxin. Among the tested strains, Streptococcus agalactiae was found to be sensitive to all the toxins, with the exception of OTA. T-2 toxin and FB1 were the most effective in slowing down the growth of Staphylococcus aureus. AFB1 affected the growth of Yersinia enterocolitica. The growth rate of Escherichia coli and Salmonella infantis was decreased by FB1. Among the bacterial strains used in this study, only the growth of Erysipelothrix rhusiopathiae was inhibited by OTA. Thus, using appropriate tester strains it should be possible to set up a broad-range microtubidimetry assay for individual mycotoxin screening in vitro. We concluded that the microtitration technique provides a rapid, convenient and high-throughput capacity system to analyse bacteria-mycotoxin interactions.

Effects of tamoxifen, melatonin, coenzyme Q10, and L-carnitine supplementation on bacterial growth in the presence of mycotoxins.

The involvement of toxic oxygen intermediates in the bacteriostatic effects of mycotoxins (T-2 toxin, deoxynivalenol, ochratoxin A, aflatoxin B1, and fumonisin B1) was investigated by producing bacterial growth curves using turbidimetry assays in the presence and absence of oxygen radical-scavenging substances. The strains used in this study included Escherichia coli (FT 101), Streptococcus agalactiae (FT 311, FT 313, FT 315), Staphylococcus aureus (FT 192), Yersinia enterocolitica (FT 430), Salmonella infantis (FT 431), Erysipelothrix rhusiopathiae (FT 432), Lactobacillus plantarum (FT234) and Lactobacillus casei (FT 232). Tamoxifen, melatonin, l-carnitine and coenzyme Q10 were used as radical scavengers against oxygen toxicity to the strains studied. Tamoxifen was the most effective in inhibiting bacterial growth when used at a high concentration, whereas melatonin and l-carnitine were less effective. A combination of l-carnitine and coenzyme Q10 provided better protection against oxygen toxicity caused by the mycotoxins growth than they did individually. It was concluded that oxygen radicals are involved in the killing of bacteria and that there is endogenous formation of toxic oxygen products by mycotoxins. The objective of this study was to determine whether the antioxidants were able to counteract the toxic effects of the mycotoxins. The data obtained indicate that bacterial growth can be inhibited especially by T-2 toxin, aflatoxin B1 and ochratoxin A and that this effect can be partially counteracted by antioxidants such as coenzyme Q10 plus l-carnitine.

T-2 toxin-induced DNA damage in mouse livers: the effect of pretreatment with coenzyme Q10 and alpha-tocopherol.

Active oxygen species are reported to cause organ damage. This study was therefore designed to determine whether oxidative stress contributed to the initiation or progression of hepatic DNA damage produced by T-2 toxin. The aim of the study was also to investigate the behaviour of the antioxidants coenzyme Q10 (CoQ10), and alpha-tocopherol (vitamin E) against DNA damage in the livers of mice fed T-2 toxin. Treatment of fasted mice with a single dose of T-2 toxin (1.8 or 2.8 mg/kg body weight) by oral gavage led to 76% hepatic DNA fragmentation. T-2 toxin also decreased hepatic glutathione (GSH) levels markedly. Pretreatment with CoQ10 (6 mg/kg) together with alpha tocopherol (6 mg/kg) decreased DNA damage. The CoQ10 and vitamin E showed some protection against toxic cell death and glutathione depletion caused by T-2 toxin. Oxidative damage caused by T-2 toxin may be one of the underlying mechanisms for T-2 toxin-induced cell injury and DNA damage, which eventually lead to tumourigenesis.

Evaluation of the possible role of coenzyme Q10 and vitamin E in juvenile neuronal ceroid-lipofuscinosis (JNCL).

The juvenile type of neuronal ceroid lipofuscinosis (JNCL) is a recessively inherited, progressive neurodegenerative disease. In this study the levels of the antioxidant factors coenzyme Q10 (CoQ10) and vitamin E (alpha-tocopherol) were measured in plasma samples of 29 JNCL patients and compared to 48 healthy controls. A significant reduction of the coenzyme Q10 level (0.59 +/- 0.25 microgram/ml) was observed in JNCL patients when compared to control subjects (0.80 +/- 0.26 microgram/ml). The level of vitamin E was also reduced markedly in JNCL patients when compared to controls (10.4 +/- 4.1 and 12.1 +/- 4.5 micrograms/ml, respectively). The low levels of CoQ10 and vitamin E in JNCL patients may indicate an impaired antioxidant protection in this disease.

Association of cadmium with human breast cancer.

The carcinogenic potential of cadmium might be affected by several factors such as smoking, hormones and presence of other metals, such as selenium and zinc. Cadmium was analyzed in breast-fat tissue of 43 breast cancer patients and 32 healthy control subjects. Patients were thoroughly characterized according to such variables as stage of cancer, smoking habits, and number of children. Correlation of cadmium levels with these variables, with hormone receptors, and with previously reported selenium and zinc were all analyzed. The mean cadmium concentration found in breast cancer patients (20.4 +/- 17.5 micrograms/g) did not differ significantly from that of the healthy controls (31.7 +/- 39.4 micrograms/g). However, unexpectedly high concentrations of cadmium (3.2-86.9 vs. 0.1-160.4 micrograms/g) were found in breast samples, which may indicate that cadmium binding proteins exist in human breast tissue. Correlation of cadmium with smoking rate of cancer patients was positive (Rs = 0.0505, p < 0.05). Correlation of cadmium with estrogen receptors in breast cancer was suggestive (Rs = 0.309, 28 cases, P = 0.06). No correlation was found with other trace elements such as selenium, zinc and copper. These results seem neither to prove nor to disprove the role of cadmium in breast cancer initiation, promotion or progression.

Plasma zinc and copper concentrations in rheumatoid arthritis: influence of dietary factors and disease activity.

Forty patients with rheumatoid arthritis performed a thorough 7-d diet recording. The food diaries were analyzed together with clinical and laboratory data by means of stepwise multiple linear regression to clarify the effects of both diet and the inflammatory disorder on the plasma concentrations of zinc and copper. The patients' daily dietary intakes of zinc and copper (24.3 +/- 7.54 and 3.48 +/- 1.55 mg/MJ) were comparable to the corresponding intakes in the ordinary Finnish diet (24.0 and 3.68 mg/MJ). In multivariate analyses the best predictors of plasma trace elements were the measures of disease activity and not the dietary factors. As an exception to this, there was a strong correlation between plasma copper-copper intake ratio and zinc intake both in univariate (r = -0.638, P less than 0.001) and multivariate analysis. This suggests that zinc depresses copper absorption with intakes in normal, physiological ranges.

Serum zinc, copper and selenium in rheumatoid arthritis.

Rheumatoid arthritis (RA) is characterized by low serum Zn and high serum Cu. In multiple linear regression both were explained by disease activity parameters. It is suggested that interleukin-1 causes both changes by 1) increasing the metallothionein-mediated hepatic uptake to serum Zn and 2) upregulating ceruloplasmin (acute phase reactant) gene and synthesis in liver and subsequently the level of ceruloplasmin-Cu complexes in the blood. Cu absorption was diminished by zinc intake. Cu- and Zn-dependent erythrocyte SOD was increased in RA. In contrast to plasma GSHPx serum selenium was low in RA and this was associated with disease activity parameters.

Altered elemental profiles in neuronal ceroid lipofuscinosis.

The elemental profiles of thrombocytes and mononuclear cells were investigated in five patients with Infantile and eight with Juvenile Neuronal Ceroid Lipofuscinosis. The patients with the infantile form had suffered from the disease for a year and those with the juvenile form for some six years. The thrombocytes exhibited increased concentrations of calcium and magnesium, but the same concentrations of iron and zinc as found in healthy subjects. The mononuclear cells exhibited an increased concentration of iron and a reduced concentration of zinc. The elevated concentrations of magnesium, calcium and iron in the thrombocytes and mononuclear cells may represent the end products of ceroid pigmentation. Five patients with Juvenile and one with Infantile Neuronal Ceroid Lipofuscinosis were treated with antioxidants along with vitamins E, B2 and B6, but this treatment did not affect significantly the concentration of iron in the mononuclear cells. However, selenium was detected in some mononuclear cells in all the patients so treated. This was unexpected since iron (III), being antagonistic to selenium in the form of selenite--which was the antioxidant given--forms a stable complex which cannot be broken down biologically.

Serum cholesterol and vitamins A and E in juvenile chronic arthritis.

Serum total cholesterol is decreased during acute infections and in adults with rheumatoid arthritis, probably partly because of enhanced lipid peroxidation. Oxidative stress also causes augmentation of inflammation and tissue damage in arthritic synovium. Therefore, concentrations of serum total cholesterol and the antioxidant vitamins A and E were studied in 125 children with juvenile chronic arthritis. Total serum cholesterol was significantly lower in the patients than in healthy children in most age groups and correlated with the markers of disease activity, haemoglobin and the erythrocyte sedimentation rate. In age- and sex-adjusted stepwise multiple linear regression, serum zinc had a significant predictive value for cholesterol. The vitamin A concentrations in the sera of the patients was virtually the same as in the healthy controls, though serum vitamin E concentrations were low (22.8 +/- 15.2 vs 30.5 +/- 4.3 mumol/l, p less than 0.001). The deficiency in vitamin E was not compensated for by another lipoperoxide antioxidant, glutathione peroxidase. Only serum cholesterol had an independent explanatory significance for vitamin E in multiple linear regression analysis (partial correlation 0.554, p less than 0.001). It is suggested that low vitamin E and impairment of the anti-oxidant protection further contribute to low serum cholesterol values in JCA.

The effect of peroral lynestrenol on serum lipids and lipoproteins in therapeutic amenorrhea of mentally retarded women.

Serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), apolipoprotein A1 (Apo A1), apolipoprotein B (Apo B), and triglyceride (TG) were measured and that of low density lipoprotein cholesterol (LDL-C) calculated in blood samples obtained from mentally handicapped women undergoing therapeutic amenorrhea (TA) induced by 5-10 mg of peroral lynestrenol, some receiving, some not receiving simultaneous anticonvulsant therapy (phenytoin, carbamazepine or barbiturate, alone or in combination). In addition, these analyses were carried out in women receiving only anti-convulsants and in controls (mentally handicapped women not receiving any of the above-mentioned medications). Significantly lower HDL-C, Apo A1, TG and cholesterol concentrations were measured in TA patients receiving lynestrenol only, than in those receiving anticonvulsants only, or in controls (p less than 0.001). With regard to HDL-C and Apo A1, patients receiving both lynestrenol and anticonvulsants were intermediate between lynestrenol only patients and controls, but the HDL-C/LDL-C and Apo A1/Apo B ratios were similar to those observed in lynestrenol only patients. Addition of 8 or 12 mg of estriol succinate to the lynestrenol regimen was virtually without an effect. However, halving of the lynestrenol dose resulted in a significant increase in HDL-C and in the HDL-C/LDL-C and Apo A1/Apo B ratios (p less than 0.001 or p less than 0.01), respectively. The lynestrenol dose was thus the most important determinant of lipoprotein pattern and should be kept as small as possible in order to reduce cardiovascular risk.

On the etiopathogenesis and therapy of Down syndrome.

The etiopathogenesis of Down syndrome is reviewed concentrating on the possible consequences of over-expression of cytoplasmic superoxide dismutase gene located in chromosome 21. Increased superoxide dismutase activity may generate free radical stress through overproduction of hydrogen peroxide. The significance of inadequate adaptive responses, i.e. increase of the selenoenzyme glutathione peroxidase activity in the central nervous system and in the thyroid gland is discussed. Suggestions are made for prevention of the progress of Down syndrome and intervention studies with antioxidant supplementation are proposed.

Serum trace elements in juvenile chronic arthritis.

We evaluated the serum concentrations of zinc, copper and selenium in 125 patients with juvenile chronic arthritis (JCA). Trace element levels showed distinct abnormalities as compared with those of a large group of healthy children. Serum zinc and selenium concentrations were lower and those of copper higher in children with arthritis than in healthy children and, further, patients with polyarthritis had significantly higher copper and lower zinc levels than those with oligoarthritis. Serum zinc levels showed a direct correlation with hemoglobin and an inverse correlation with values for the erythrocyte sedimentation rate (ESR), whereas copper correlated directly with ESR. Selenium values did not correlate with the activity of the disease, but were low in the patients with arthritis of long duration.

Comparison of the clinical courses in patients with juvenile neuronal ceroid lipofuscinosis receiving antioxidant treatment and those without antioxidant treatment.

Juvenile neuronal ceroid-lipofuscinosis (JNCL) is a progressive encephalopathy characterized by a neural and extraneural accumulation of ceroid and lipofuscin like storage cytosomes and by an autosomal recessive inheritance. It begins with a gradual loss of vision at the age of 4-7 years and is accompanied by epilepsy, a loss of motor function, and a progressive dementia (Santavuori 1988). We have studied 26 Finnish JNCL patients treated with vitamins E, B2, B6 and sodium selenite (antioxidant treatment) by using a JNCL disease specific scoring system introduced by Kohlschütter et al. (1988). Scores were given for the problems of vision, intellect, language, motor function, as well as epilepsy, and compared with the data of 17 German JNCL patients not treated with antioxidants (Kohlschütter et al. 1988). Loss of vision began at the same time among the Finnish and the German JNCL patients. However, loss of intellectual, language, and motor functions and total blindness occurred later among the group of Finnish JNCL patients treated with antioxidants. Courses of the epileptic seizures were rather heterogenous and slightly favouring the Finnish patients. This study supports the theory that antioxidant treatment retards JNCL disease. The study design, however, contains many possible biases, so that the results must be interpreted cautiously.