Elevated levels of IL-24 in systemic lupus erythematosus patients.

OBJECTIVE: This study aimed to assess IL-24 levels and their association with clinical manifestations in patients with systemic lupus erythematosus (SLE). METHODS: There were 75 patients with SLE and 58 healthy controls recruited in this study. Serum levels of IL-24 were measured by enzyme-linked immunosorbent assays, and mRNA levels of IL-24 were tested by quantitative real-time polymerase chain reaction . The area under the curve of the receiver operating characteristic (ROC) curve was used for diagnostic ability of the inflammatory cytokine. RESULTS: Serum IL-24 levels were significantly higher in SLE patients than that in healthy controls. SLE patients with nephritis had higher IL-24 levels than those without nephritis. Active SLE patients showed higher expression of IL-24 as compared to less active disease patients. The mRNA levels of IL-24 were much higher in SLE patients. Correlation analysis showed significant correlation between serum IL-24 levels and SLE disease activity index. In addition, ROC analysis may suggest good ability of serum IL-24 in differentiating SLE. CONCLUSION: The inflammatory cytokine correlated with SLE disease activity, and may be involved in this disease pathogenesis.

LPS-evoked IL-18 expression in mesangial cells plays a role in accelerating lupus nephritis.

OBJECTIVES: Systemic lupus erythematosus is occasionally accompanied with bacterial infection. Lipopolysaccharide (LPS) from bacteria can accelerate and exacerbate lupus nephritis (LN) in animal models, but some mechanisms underlying the LPS-induced acceleration are still unclear. First, it is not known whether LPS can stimulate mesangial cells (MCs) to secrete the pro-inflammatory cytokine, interleukin (IL)-18. Second, it is also unclear whether LPS and/or IL-18 can induce MC apoptosis. Here, we attempted to clarify the cause-and-effect relationships between LPS stimulation, IL-18 production and MC apoptosis to address the above questions. METHODS: LPS was used to induce accelerated LN in LN-prone mice. LPS and IL-18 were also used to treat cultured MCs isolated from the mice. IL-18 expression and MC apoptosis were investigated by in situ hybridization, the TUNEL method, reverse transcription- polymerase chain reaction (RT-PCR), western blotting, DNA electrophoresis and flow cytometry. NFkappaB was detected by immunofluorescent staining. RESULTS: In the LPS-accelerated LN mice, we observed co-existence of IL-18 expression, hyperplasia, apoptosis, and activated apoptotic signal transduction in MCs, as well as marked neutrophil infiltration in the glomerulus, especially around the mesangial region. In cultured MCs, LPS greatly enhanced IL-18 expression, but did not induce apoptosis, while mouse IL-18 did not induce apoptosis or activate apoptotic signal transduction in MCs. CONCLUSIONS: We conclude that LPS can evoke IL-18 production in MCs, but neither LPS nor IL-18 directly induces apoptosis or activates apoptotic signal transduction in the cells. We infer that LPS-induced IL-18 production by MCs could be a mediator by which LPS accelerates and exacerbates LN.

Differences between plasma and adipose tissue biomarkers of carotenoids and tocopherols.

Biomarkers of dietary exposure or nutritional status are sought actively to overcome limitations of traditional dietary methodology. We compared plasma and adipose tissue biomarkers for carotenoids and tocopherols. The data consisted of samples from 91 men and 122 women, ages 45-70 years, from the control group of the European Community Multicentre Study on Antioxidants, Myocardial Infarction, and Cancer of the Breast (EURAMIC) Study. Pearson correlations between plasma and adipose tissue measurements for beta-carotene, lycopene, and alpha-tocopherol adjusted for smoking status displayed low, although significant, correlations of 0.39, 0.24, and 0.39, respectively. The correlation was further stratified by sex. After being corrected for measurement error using deattenuation factors obtained from a reproducibility study, the stratified correlation coefficients were as high as 0.80 for beta-carotene in men, 0.62 for lycopene in women, and 0.52 for alpha-tocopherol in women. In addition, plasma and adipose tissue measurements from the myocardial infarction (MI) subset of the EURAMIC study population were used to evaluate the odds of MI, adjusting for confounders. We found that the concentration of lycopene in plasma was not positively associated significantly with MI (odds ratio, 1.78; P = 0.26). Adipose tissue lycopene, in contrast to reports elsewhere on the total population, showed an inverse association with MI (odds ratio, 0.62; P = 0.15). These results suggest that plasma and adipose carotenoids represent different markers for nutritional status and cannot be used interchangeably in epidemiological and dietary validation studies.

The extraction and properties of superoxide dismutase from porcine blood.

Crude Superoxide dismutase (SOD) was prepared by extraction with chloroform-ethanol, centrifugation and acetone precipitation. The specific activity of the SOD was 1570 units mg(-1), and an unusual ultraviolet spectrum with maximum absorbance at 256.4 nm was observed. When the SOD was kept at 40, 50 and 60 °C for 10 min, it showed little loss of stability but it retained only 95, 69, 45 and 2.5% of its original activity after holding at 60, 70, 75, and 80 °C for 5 min., respectively. When cupric ions were added, the thermostability of SOD was enhanced. When the extracted SOD was kept at -23, 4, 25 and 30 °C for 20 weeks, the relative activity retained was 76, 67, 26 and 16%, respectively. Using the sensitive test with cyanide and hydrogen peroxide, the acrylamide gel of the SOD showed this enzyme was of the Cu Zn type. The molecular weights of the holoenzyme and its subunits, estimated by SDS-polyacrylamide gel electrophoresis, were approximately 31 and 15 kD, respectively.

Organic solvent-soluble lipofuscin pigments and glutathione peroxidase in mouse brain and heart: effects of age and vitamin E.

The effect of age and dietary supplementation of vitamin E or N,N'-diphenyl-p-phenylenediamine (DPPD) on organic solvent-soluble lipofuscin pigments (OLP) and glutathione peroxidase (GSH-Px) activity in mouse heart and brain was investigated. Four groups of 32 female weanling mice were fed a basal diet containing either 0, 30 or 300 ppm RRR-alpha-tocopheryl acetate (d-alpha-tocopheryl acetate) or 30 ppm DPPD from 2 to 18 mo of age. Neither GSH-Px activity nor dietary supplementation of vitamin E or DPPD had an effect on OLP concentrations in the brain or heart. OLP levels were two- to fourfold higher at 12 mo of age in the heart and were lower at 18 mo of age in the brain than at 2 or 9 mo of age. GSH-Px activity increased with age in the heart tissue of vitamin E-deficient and DPPD-supplemented mice only. No change in GSH-Px activity was observed in the brain due to diet or increasing age. These results suggested that OLP concentrations were not affected by dietary supplementation of vitamin E or DPPD but were affected by age-related factors in the mouse brain and heart.

Effect of selenite, vitamin E and N,N'-diphenyl-p-phenylenediamine on liver organic solvent-soluble lipofuscin pigments in mice.

The present experiment was designed to investigate the effect of selenium (Se) supplementation, as sodium selenite, on organic solvent-soluble lipofuscin pigment (OLP) accumulation and glutathione peroxidase (GSH-Px) activity in the livers of mice fed varying levels of vitamin E or N,N'-diphenyl-p-phenylenediamine (DPPD). Four groups of 16 female, weanling mice each were fed either a vitamin E-deficient diet, a diet supplemented with 30 mg/kg or 300 mg/kg vitamin E (as RRR-alpha-tocopheryl acetate), or a diet supplemented with 30 mg/kg DPPD. Each diet contained 0.05 ppm Se. At 5 months of age, eight animals from each dietary group were supplemented with an additional 0.1 ppm Se, as sodium selenite, in their drinking water. The remaining animals were fed their original diets through the 9-month experimental period. Selenite supplementation resulted in a significant increase in OLP concentration and GSH-Px activity in the liver of mice fed vitamin E- or DPPD-supplemented diets. Normal levels of vitamin E and DPPD (30 mg/kg) were not sufficient to protect against the oxidative effects of selenite; however, 10 times the normal level of vitamin E (300 mg/kg) markedly suppressed this oxidative effect.

A comparison of copper-loading disease in Bedlington terriers and Wilson's disease in humans.

Eleven Bedlington terriers were found to have a mean hepatic copper concentration of 6,321 micrograms/g dry wt (normal, 200 micrograms/g dry wt) and renal copper concentration that was three or four times normal. Brain copper levels were normal in younger dogs, were elevated in two older dogs, and were 100 times normal in one dog that died of the disease. Increased concentrations of copper in the liver, kidney, and brain also characterize Wilson's disease. Erythrocyte survival was normal in three affected dogs, but serum glutamic-pyruvic transaminase levels were usually elevated. Unlike the hypoceruloplasminemia of patients with Wilson's disease, plasma ceruloplasmin activity was not only normal but was also slightly elevated in the terriers. Despite their normal or excessive ceruloplasmin, the Bedlington terriers could convert ionic 64Cu to radioceruloplasmin but did so only very slowly. These dogs accumulated significantly more 64Cu in their livers than normal, much like patients with Wilson's disease do before symptoms develop.

Lectins and the soybean-Rhizobium symbiosis. I. Immunological investigations of soybean lines, the seeds of which have been reported to lack the 120 000 dalton soybean lectin.

Seeds of six soybean lines (Glycine max (L.) Merr. cv. Columbia, D68-127, Norredo, Sooty, T-102, Wilson 5) have been reported to lack the 120 000 dalton soybean lectin. Immunodiffusion and radioimmunoassay using anti-soybean lectin immunoglobulin failed to detect the lectin in seeds of five lines, but D68-127 seeds contained as much soybean lectin as the control line, Harosoy 63. The D68-127 seed lectin could be purified by affinity chromatography on Sepharose-N-caproylgalactosamine, and was indistinguishable from the conventional soybean lectin by the following criteria: electrophoretic migration in acidic and alkaline buffers, subunit molecular weight and composition, analytical isoelectric focusing, gel filtration chromatography. Phosphate-buffered saline extracts of roots, hypocotyls, stems, and leaves of 3--66-day-old Norredo and Harosoy 63 plants lacked soybean lectin, as determined by hemagglutination and radioimmunoassay (detection limit: 1.4 micrograms soybean lectin/g dry weight tissue). Cotyledons of Harosoy 63 (but not Norredo) contained large quantities of the lectin, which diminished as the plants aged. 5-day-old roots and hypocotyls of 20 soybean lines did not contain soybean lectin. Roots of Columbia, Norredo, Sooty, T-102, Wilson 5, and Harosoy 63 (control) were nodulated by a variety of strains of Rhizobium japonicum and Rhizobium sp.

Effect of dietary vitamin E and aging on tissue lipofuscin pigment concentration in mice.

This study reports a re-investigation of the effect of dietary vitamin E upon tissue organic solvent soluble lipofuscin pigment concentrations. Female weanling mice were fed a vitamin E deficient, vitamin E or N,N'-diphenyl-phenylenediamine (DPPD) supplemented diet up to 18 months of age. Lipofuscin concentrations were measured by a quantitative method which is based on fluorescence spectroscopy. Of all tissues measured (uterus, lung, spleen, kidney, liver, heart and brain), only the liver responded and showed lower pigment concentrations due to vitamin E treatment. In addition, in the liver, up to 12 months of age, vitamin E supplementation resulted in gradually decreasing pigment concentrations, but by 18 months of age, pigment concentrations were increased by 5 to 10 times in all diet groups. The effect of DPPD was similar to vitamin E. Tissue lipofuscin pigment concentrations in 18-month-old mice were lowest in the uterus and highest in the heart. The data indicate the possibility of a turnover of the organic solvent soluble lipofuscin pigments in the liver.