The Effect of Cyclosporine A on Dermal Fibroblast Cell - Transcriptomic Analysis of Inflammatory Response Pathway.

BACKGROUND: The first immunosuppressive drug - cyclosporine A (CsA) has many unquestioned merits in maintaining organ transplants in patients, as well as, in the treatment of many inflammatory diseases, also associated with cutaneous manifestations. The main task of this drug is to suppress the inflammatory response at the sites of action, which is not well known. OBJECTIVE: The objective of this study was to evaluate the influence of CsA in therapeutic concentration on the expression of genes associated with the inflammatory response pathway in normal human dermal fibroblasts (NHDF; CC-2511), and this study attempted to determine the mechanism of its action. METHODS: The cytotoxicity MTT test was performed. The expression of the inflammatory response pathway genes was determined using HG-U133A_2.0 oligonucleotide microarrays. Statistical analysis was performed by GeneSpring 13.0 software using the PL-Grid platform. RESULTS: Among the 5,300 mRNA, only 573 were changed significantly in response to CsA compared to the control fibroblasts (P≤0.05). CsA inhibited the expression of most genes associated with the inflammatory response in NHDFs. There were only 19 genes with a fold change (FC) lower than -2.0, among which EGR1, FOS, PBK, CDK1 and TOP2A had the lowest expression, as did CXCL2 which can directly impact inflammation. Furthermore, ZNF451 was strongly induced, and COL1A1, COL3A1, IL33, TNFRSFs were weakly up-regulated (FC lower than 2.0). CONCLUSION: The CsA in therapeutic concentration influences the genes linked to the inflammatory response (in the transcriptional level) in human dermal fibroblasts. The findings suggest that the potential mechanism of CsA action in this concentration and on these genes can be associated with a profibrotic and proapoptotic, and genotoxic effects.

Influence of static magnetic fields up to 700 mT and dihydrochalcones on the antioxidant response in fibroblasts.

The effects of a static magnetic field (SMF) and the dihydrochalcones phloretin and phloridzin on the redox homeostasis of fibroblasts were investigated. The aim of the present study was to determine the redox homeostasis of fibroblasts that were simultaneously exposed to a static magnetic field and the dihydrochalcones phloretin and phloridzin. The fibroblasts were cultured for 72 h in special magnetic test chambers at different moderate intensities (0.4, 0.55 and 0.7 T). In this report, the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione transferase (GST); the concentrations of malondialdehyde (MDA), adenosine triphosphate (ATP) and total antioxidant status were measured using commercially available kits. We did not observe any impairment in the redox balance in cells in fibroblasts that were only exposed to static magnetic fields of different intensities or In fibroblast cultured with dihydrochalcones and exposed to static magnetic field increase the SOD, GPx, GST activities and MDA concentration. Our investigations revealed that the activities of SOD, GPx, GST and the concentration of MDA that were determined for the fibroblasts that were cultured with dihydrochalcones were higher in the presence of a static magnetic field. Our results indicated that exposure to SMF (0.7 T) with dihydrochalcones induces oxidative stress in fibroblasts.

Studies on the effect of quercetin and nitrates on the redox homeostasis using in vitro model.

Antioxidants are widely considered to be a preventive measure for many diseases and beneficial for health. However, an increasing number of reports suggest a lack of any influence by antioxidants on health or even harmful pro-oxidative effects of antioxidants. In most cases, the research was conducted with respect to a chosen antioxidant, without considering the presence of other chemical substances present in food, with which these compounds may react. The aim of this work was to determine whether and to what extent the simultaneous presence of quercetin and sodium nitrate influences oxidative-reductive homeostasis in fibroblast cultures. Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and nitric oxide synthase (NOS) activities were measured together with nitric oxide (NO) concentration and total antioxidant status (TAS). An increase in the activity of all the enzymes measured and in the NO concentration was determined compared with the control culture. The most prominent changes were observed at the highest quercetin concentration. These results indicate that the simultaneous presence of quercetin and sodium nitrate disrupts the oxidative-reductive homeostasis in fibroblasts.

N-acetylcysteine protects against fluoride-induced oxidative damage in primary rat hepatocytes.

Fluoride induces the overproduction of free radicals, which might in turn affect various biochemical parameters. Therefore, the aim of this study was to elucidate the role of N-acetylcysteine (NAC) in decreasing fluoride-induced oxidative stress. The fluoride intoxicated (0.002; 0.082; 0.164mmol/l) rat hepatocytes was pre-treated (60min) and simultaneously treated with NAC (1mmol/l). The resulting levels of lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and intracellular reduced glutathione (GSH) were measured along with the total antioxidant status (TAS) to determine whether NAC treatment reduced cell damage and/or the antioxidant state. These results suggest that NAC pre-treatment provides protection against fluoride-induced oxidative stress in hepatocytes.

Effect of a static magnetic fields and fluoride ions on the antioxidant defense system of mice fibroblasts.

The results of studies on the biological influence of magnetic fields are controversial and do not provide clear answers regarding their impact on cell functioning. Fluoride compounds are substances that influence free radical processes, which occur when the reactive forms of oxygen are present. It is not known whether static magnetic fields (SMF) cause any changes in fluoride assimilation or activity. Therefore, the aim of this work was to determine the potential relationship between magnetic field exposure to, and the antioxidant system of, fibroblasts cultured with fluoride ions. Three chambers with static magnetic fields of different intensities (0.4, 0.6, and 0.7 T) were used in this work. Fluoride ions were added at a concentration of 0.12 mM, which did not cause the precipitation of calcium or magnesium. The results of this study show that static magnetic fields reduce the oxidative stress caused by fluoride ions and normalize the activities of antioxidant enzymes, including superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). Static magnetic fields modify the energy state of fibroblasts, causing an increase in the ATP concentration and a decrease in the MDA concentration. These results suggest that exposure to fluoride and an SMF improves the tolerance of cells to the oxidative stress induced by fluoride ions.

Application of EPR Spectroscopy to Examination of the Effect of Sterilization Process on Free Radicals in Different Herbs.

Free radicals in the original and sterilized caraway, curry, curcuma and cardamom were studied. An X-band (9.3 GHz) electron paramagnetic resonance (EPR) spectroscopy was the experimental technique. Effect of microwave power in the range of 2.2-70 mW on amplitudes, linewidths, and lineshape parameters of the EPR spectra was tested. Free radicals concentrations in the non- and sterilized herb samples were compared. The aim of this work was to determine properties and concentration of free radicals in steam sterilized caraway, curry, curcuma and cardamom. It was pointed out that free radicals (~1018 spin/g) exist in both the original and sterilized herbs. Complex free radical system with oxygen and carbon paramagnetic centers characterizes the examined herbs. Homogeneously dipolar broadened EPR spectra were measured for all the tested herbs. Slow spin-lattice relaxation processes exist in the examined samples. Practical usefulness of EPR method in food technology was discussed.

Cathepsin D activity in nasal polyps.

OBJECTIVES: The aim of the study was to assess the activity of cathepsin D in polyps removed during first-time FESS surgery and in recurrent polyps removed during successive FESS surgeries. DESIGN AND METHODS: The study examined 24 polyps: 11 polyps were removed during first-time surgical procedures (termed primary polyps in the study), and 13 recurrent polyps were removed in successive FESS surgeries (termed polyps after re-polypectomy in the study). The activity of cathepsin D was determined by measuring the tyrosine released from denatured hemoglobin. RESULT: The average cathepsin D activity in polyps after re-polypectomy was 74% higher than the average activity in primary polyps, whereas the difference between the maximum and the minimum cathepsin D activities in polyps after re-polypectomy was twofold greater than the respective difference in primary polyps. CONCLUSION: Differences in cathepsin activity and the protein content, likely to be significant in the process of polyp recurrence, were observed in both groups of polyps.

The influence of small doses of paracetamol on rabbit liver.

Paracetamol has properties that make it useful in many kinds of drugs that are readily available to users. These drugs are often used for extensive periods of time without a doctor's supervision, which might lead to unexpected overdose. The aim of this work was to assess the extent of liver damage in rabbits after administering them with paracetamol for approximately 2 months. The results we obtained suggest that paracetamol administered for a long time causes liver function impairment. It was, however, not possible to determine the impairment by measuring the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT) or gamma-glutamylotranspeptidase (GGTP). It was determined by measuring the concentration of ketone bodies and arterial ketone body ratio (AKBR) value. Even when taken in small doses, paracetamol causes changes in liver metabolism when taken for an extended period of time.

[Urine iodine excretion by healthy pregnant women after vitamin and microelement supplementation (Materna) in the first trimester]. / Wydalanie jodu z moczem u zdrowych kobiet rozpoczynajacych stosowanie preparatu Materna w pierwszym trymestrze ciazy.

OBJECTIVES: The aim of the study was the assessment of iodine turnover in healthy pregnant women supplemented by microelement and vitamin preparation Materna. METHODS: To assess iodine supplementation, a modified PAAM method was used for measurement of iodine concentration in morning urine. MATERIALS: 50 pregnant healthy women were tested for Materna supplementation. Urine iodine concentration was measured starting from the I trimester of pregnancy and then after 1, 2 and 3 months of Materna administration. RESULTS: It has been found, that 3 months of supplementation with Materna reduces iodine deficiency in 83% of primiparas, while in the same time urine iodine concentration was normal in only 30% of women in their third and next pregnancy. CONCLUSION: In our opinion iodine urine concentration should be assessed in all multiparas using Materna after 1 month of supplementation in order to correct individual iodine supplementation doses.