Reactive oxygen species and serum antioxidant defense in juvenile idiopathic arthritis.

In autoimmune inflammatory diseases, including juvenile idiopathic arthritis (JIA), which leads to joint destruction, there is an imbalance between production of reactive oxygen species (ROS) and their neutralization which, as a consequence, leads to "oxidative stress." The aim of the study was to assess the concentration of oxidative stress markers: nitric oxide (NO), a degree of lipid membrane damage, and total antioxidant plasma capacity in children with JIA. Thirty-four children with JIA were included into the study. A degree of lipid membrane damage (lipid peroxidation products) was estimated as thiobarbituric acid-reactive substances (TBARs), NO concentration as NO end-products: nitrite/nitrate (NO2(-)/NO3(-)) and total antioxidant plasma capacity as ferric reducing ability of plasma (FRAP). NO2(-)/NO3(-) serum concentration in children with JIA was statistically significantly higher than that in healthy children (p = 0.00069). There was no significant difference in TBAR levels between children with JIA and the control group. FRAP in sera of children with JIA was lower than that in healthy children, but the difference was not statistically significant. A statistically significant positive correlation was observed between NO end products and the 27-joint juvenile arthritis disease activity score (JADAS-27) and ESR, and a negative correlation was observed between FRAP and C-reactive protein (CRP) and white blood cell count (WBC). Our results confirm the increased oxidative stress in children with JIA. Overproduction of NO and decrease in the antioxidant plasma capacity may be involved in JIA pathogenesis.

Response to fasting in an unnaturally obese carnivore, the captive European polecat Mustela putorius.

The European polecat (Mustela putorius) is a naturally lean carnivore prone to excessive weight gain in captivity. This study assessed its suitability to investigate the natural history of the obese phenotype displayed in overweight humans, domestic animals, and seasonally obese wild mammals. Ten farm-bred polecats were subjected to a 5-day fast with 10 controls. Obesity (40% body fat) was associated with an unfavorable plasma lipid profile and high glucose and insulin concentrations. The polecats were in phase II of fasting with normoglycemia, low liver carbohydrate stores, and decreased plasma concentrations of urea and most amino acids. Although the plasma nonesterified fatty acid (NEFA) levels were elevated, the adipose tissue lipase activities suggested a blunted lipolytic response. Lipid mobilization was more efficient from intraabdominal fat. The animals developed hepatic lipidosis with elevated NEFA influx into the liver and losses of n-3 polyunsaturated fatty acids and arginine as hypothetical etiological factors. The plasma leptin, insulin, and triiodothyronine levels decreased but were not accompanied by reduced sex steroid or increased stress hormone concentrations. The blunted lipolytic response often encountered in obesity suggests that the organism is trying to defend the obese phenotype. Liver lipidosis and decreased insulin and triiodothyronine levels seem to be among the most consistent responses to fasting manifested in diverse mammalian orders and different levels of body fatness. The polecat could be recommended as an easily accessible carnivorean model to study the natural history of the obese phenotype and its comorbidities.

Effects of preservation methods on stable isotope signatures in bird tissues.

Increasing use is being made of stable isotopes as indicators of habitat use and trophic ecology of animals. Preservation of tissues can alter stable isotope signatures. We investigated the effects of addition of ethanol and NaCl solution (hereafter 'salt'), and of freezing and drying, on carbon and nitrogen isotopic values in blood of the spectacled petrel Procellaria conspicillata, and compared these with those from simultaneously growing feathers. The mean delta(13)C values of blood preserved in ethanol was significantly higher, and of blood preserved in salt was significantly lower than that of dried or frozen samples. delta(13)C values in ethanol showed high variation according to brand and batch and could account for the differences found in delta(13)C ratios in ethanol-preserved blood samples. Mean delta(13)C and delta(15)N values in growing feathers were higher than in blood, suggesting tissue-specific fractionation. We conclude that different methods of preserving tissues such as blood may bias stable isotope values, and urge researchers to consider this issue. Air drying is proposed as a practical and unbiased method for blood preservation in field situations where freezing is not a practical option, and a mathematical approach is suggested to permit comparison between studies using different preservation methods or tissues.

Establishing the use of melatonin as an adjuvant therapeutic against paraquat-induced lung toxicity in rats.

It is well known that the intake of paraquat (PQ) causes severe tissue injury leading to numerous fatalities. Considering that the main target for PQ toxicity is the lung and involves the production of reactive oxygen and nitrogen species, transcription factors and inflammatory cytokines, it may be hypothesized that the combination of a potent antiinflammatory and antioxidant agent may counteract more of PQ's effects than an antiinflammatory agent alone. For this purpose, combination of dexamethasone (Dex) and melatonin (Mel) was compared with Dex alone. A total of 40 male Wistar albino rats were divided into four groups as control, PQ, Dex only, and Dex plus Mel. The animals were given intraperitoneally a toxic dose of 19 mg/kg PQ dissolved in 1 ml saline. Control animals were injected with the same amount of saline only. A dose of 1 mg/kg Dex was administered 2 hrs after PQ administration. In the combination treatment group, 20 mg/kg Mel was given with Dex. All drugs were given every 12 hrs for a total of six doses. Five animals in PQ group and three animals in Dex only group died by the end of the study. No deaths occurred in the Dex+Mel group. Dex exerted improvements in several oxidative and antioxidative parameters. However, combination treatment provided beneficial effects against PQ toxicity far greater than Dex alone. This difference was also apparent when tissues were histologically compared. In conclusion, Mel exhibited strong additive beneficial effects with Dex and can be considered as a safe treatment modality against PQ toxicity.

Mutual changes of thioredoxin and nitrosothiols during biliary cirrhosis: results from humans and cholestatic rats.

UNLABELLED: Cholestasis is associated with changes in NO metabolism and thiol oxidation. Thioredoxin contributes to regulate vascular tone and intracellular redox status by cleaving nitrosothiols and maintaining -SH groups. This study investigated the changes in circulating thioredoxin and nitrosothiols and the relationship with protein sulfhydryls (PSH), hepatic concentrations, hyaluronate, and histology in patients with primary biliary cirrhosis (PBC) and in rats with bile duct ligation (BDL). PSH in erythrocytes were significantly decreased in stage III and IV PBC and at day 10 after BDL. Compared with controls, erythrocyte thioredoxin levels were higher in stage I through III PBC and lower in stage IV patients. Serum thioredoxin levels were significantly higher in PBC stages I and II and lower in stages III and IV. Serum nitrosothiols were higher in all PBC patients and inversely related to thioredoxin and hyaluronate. In rats, serum, hepatic, and mitochondrial thioredoxin had initially increased after BDL (day 1-3) and then decreased. After day 7 BDL, nitrosothiols were 10-fold increased in serum and liver, and even higher in mitochondria. In the liver, thioredoxin was inversely related to both nitrosothiols and PSH. In rats, the difference in time average changes from baseline among serum, hepatic, and erythrocyte thioredoxin suggests that most of circulating thioredoxin originates from the liver. CONCLUSION: Our findings indicate that cholestasis is associated with significant mutual and interrelated changes between circulating and hepatic thioredoxin and nitrosothiols. The increase of hepatic, mitochondrial, and circulating nitrosothiols with ongoing cholestasis suggests an active participation of NO in both liver injury and extrahepatic changes.

Splanchnic metabolism of nitrogenous compounds and urinary nitrogen excretion in steers fed alfalfa under conditions of increased absorption of ammonia and L-arginine supply across the portal-drained viscera.

Effects of increased ammonia and/or arginine absorption across the portal-drained viscera (PDV) on net splanchnic (PDV and liver) metabolism of nitrogenous compounds and urinary N excretion were investigated in six catheterized Hereford x Angus steers (501 +/- 1 kg BW) fed a 75% alfalfa:25% (as-fed basis) corn-soybean meal diet (0.523 MJ of ME/[kg BW(0.75).d]) every 2 h without (27.0 g of N/kg of dietary DM) and with 20 g of urea/kg of dietary DM (35.7 g of N/kg of dietary DM) in a split-plot design. Net splanchnic flux measurements were obtained immediately before beginning and ending a 72-h mesenteric vein infusion of L-arginine (15 mmol/h). For 3 d before and during arginine infusion, daily urine voided was measured and analyzed for N composition. Feeding urea increased PDV absorption (P < 0.01) and hepatic removal (P < 0.01) of ammonia N, accounting for 80% of increased hepatic urea N output (P < 0.01). Numerical increases in net hepatic removal of AA N could account for the remaining portion of increased hepatic urea N output. Arginine infusion increased hepatic arginine removal (P < 0.01) and hepatic urea N output (P < 0.03) and switched hepatic ornithine flux from net uptake to net output (P < 0.01), but numerical changes in net hepatic removal of ammonia and AA N could not account fully for the increase in hepatic urea N output. Increases in urine N excretion equaled quantities of N fed as urea or infused as arginine. Estimated salivary urea N excretion was not changed by either treatment. Urea cycle regulation occurs via a complex interaction of mechanisms and requires N sources other than ammonia, but the effect of increased ammonia absorption on hepatic catabolism of individual AA in the present study was not significant.

Changes in some nitrogenous compounds in the blood and tissues of freshwater pikeperch (Sander lucioperca) during salinity acclimation.

The effect of ambient salinity changes (0.9, 6 and 12 psu) on the levels of dissolved ammonia (DA), ninhydrin positive substances (NPS), trimethylamine (TMA) and trimethylamine oxide (TMAO) in the blood and tissue of medium-acclimated Sander lucioperca L. (also Stizostedion lucioperca) were investigated. In freshwater, blood and tissue total free amino acid levels (measured as NPS) were 3.62 mM and 60.61 mM, respectively. The NPS content increased significantly (P<0.05) in the tissue and blood on acclimation to 6 and 12 psu salinities. The mass-specific tissue TMAO concentration of pikeperch acclimated to normal freshwater was 0.413+/-0.084 micromol TMAO g(-1). Results reveal that TMAO levels are positively influenced by the external salinity medium where significant differences in mean levels occurred between the groups (P<0.05). The calculated p[NH(3)] and [NH(4)(+)] gradients reveal that the [NH(3)] gradient was consistently low (cf. the [NH(4)(+)] gradient). The gradient of p[NH(3)] decreased with the medium increased salinities. The results suggest that freshwater pikeperch may be able to resist salinity changes by manipulation of nitrogen metabolism. Free amino acids and TMAO are involved in mediating response to salinity exposure in freshwater pikeperch.

Preoperative glutamine administration induces heat-shock protein 70 expression and attenuates cardiopulmonary bypass-induced inflammatory response by regulating nitric oxide synthase activity.

BACKGROUND: Heat-shock protein 70 (HSP70) plays a major role in the pathophysiology of inflammation, and the induction of HSP70 before the onset of inflammation can reduce organ damage through a self-protective system. Glutamine is known to be an inducer of HSP70, and its preoperative administration seems useful in attenuating cardiopulmonary bypass (CPB)-induced inflammatory response. METHODS AND RESULTS: Adult male Sprague-Dawley rats (group G, received 100 mg/kg of glutamine via the right jugular vein 3 times per day for 1 week and just before the initiation of CPB; group C served as control) underwent CPB (60 minutes, 100 mL/kg per minute, 34 degrees C) and were killed 3 hours after the termination of CPB. Group G showed significantly lower plasma concentrations of interleukin-6 and interleukin-8 after CPB termination. Myocardial and respiratory damages were significantly attenuated in group G, as evidenced by Langendorff perfusion, respiratory index, and neutrophil adherence. HSP70 expressions in the heart, lung, and liver were detected only in group G before CPB and were markedly stronger in group G 3 hours after CPB termination. Although plasma nitrate+nitrite concentrations were not significantly different between the groups, endothelial-constitutive nitric oxide synthase (NOS) activity was markedly preserved and inducible NOS activity was markedly attenuated in the tissues of group G. CONCLUSIONS: These results suggest that preoperative glutamine administration induces HSP70 expression before CPB and attenuates CPB-induced inflammation by regulating NOS activity, which may be a prospective management for conferring tolerance to CPB-induced inflammatory response through a self-protective mechanism.

The detection and identification of quaternary nitrogen muscle relaxants in biological fluids and tissues by ion-trap LC-ESI-MS.

Quaternary nitrogen muscle relaxants pancuronium, rocuronium, vecuronium, gallamine, suxamethonium, mivacurium, and atracurium and its metabolites were extracted from whole blood and other biological fluids and tissues by using a solid-phase extraction procedure. The extracts were examined by using high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). The drugs were separated on a ODS column in a gradient of ammonium acetate buffer (pH 5.0) and acetonitrile. Full-scan mass spectra of the compounds showed molecular ions, and MS-MS spectra showed fragments typical of the particular compounds. LC-ESI-MS allowed an unequivocal differentiation of all muscle relaxants involved. The method was applied in a case of rocuronium and suxamethonium administration in a Caesarian section and in a case of intoxication by pancuronium injection. In both cases, the administered drugs could be detected and identified in the supplied samples.

Dietary protein, urea nitrogen appearance and total nitrogen appearance in chronic renal failure and CAPD patients.

This study was carried out to examine the mathematical relationships between the urea nitrogen appearance (UNA), total nitrogen appearance (TNA) and dietary nitrogen intake (DNI) in patients with chronic renal failure. Studies were conducted in 20 nondialyzed patients with advanced chronic renal failure (CRF) who were fed 27 constant protein diets for 24.8 +/- 9.5 days (SD) and eight patients undergoing continuous ambulatory peritoneal dialysis (CAPD), who ingested 13 constant protein diets for 20.3 +/- 4.9 days. All patients lived in a hospital research ward throughout the study and underwent full nitrogen balance measurements. Data were analyzed after patients attained equilibrium or near equilibrium with each dietary protein intake. In the CRF patients, using the mean values obtained during the equilibrium phase, there was a direct and precise correlation between the TNA and UNA, where TNA g/day = 1.19 UNA g/day + 1.27 g/day, r = 0.948. The correlation between DNI and UNA was also highly significant, but less precise, with somewhat greater 95% confidence intervals: DNI g/day = 1.20 UNA g/day + 1.74 g/day, r = 0.865. The relationship between DNI and TNA was not much more precise: DNI g/day = 0.97 TNA g/day + 0.65 g/day, r = 0.880. With the CAPD patients, the relationships were as follows: TNA g/day = 0.94 UNA g/day + 5.54 g/day, r = 0.956; DNI g/day = 0.97 UNA g/day + 6.80 g/day, r = 0.705; DNI g/day = 1.07 TNA g/day + 0.63 g/day, r = 0.760. For the CAPD patients, the lowest 95% confidence intervals were also found for the correlation between TNA and UNA. Thus, in both CRF and CAPD patients, the TNA is highly and precisely correlated with the UNA. The DNI is also significantly correlated with UNA and TNA, but the relationship is less precise. In both of these two groups of patients, the difference between the regression equations for TNA versus UNA and DNI versus UNA was, to a substantial degree, accounted for by the intercept.

[The influence of rumen metabolism on cyclic N compounds in the blood plasma of ruminants]. / Der Einfluss des Pansenstoffwechsels auf zyklische N-Verbindungen im Blutplasma von Wiederkäuern.

The purpose of this paper was to study the effect of rumen metabolism on the concentration of certain cyclic N-compounds in blood plasma. The plasma from two groups of lambs in the phase of rumen development during early and late weaning was used. As a second model, samples from steers fed at maintenance and after emptying of the rumen via cannula were used. Plasma samples were obtained from the jugular vein in regular intervals. The HPLC-technique was employed as analytical method. The plasma levels of allantoin and of hippuric acid increased about 2.2-3.7 fold and 2.4-6.4 fold in lambs during rumen development. They decreased to about 43% and 13% after emptying the rumen of steers. These metabolites appear to reflect rumen protein synthesis and fibre breakdown and are considered useful estimators of rumen function. Cytidine, uridine, uric acid and hypoxanthine were only influenced in part by rumen metabolism. Plasma pseudouridine was decreased and creatinine was increased in lambs in relation to body mass and remained unaffected in steers.

Short-term space flight on nitrogenous compounds, lipoproteins, and serum proteins.

Biochemical variables in blood were measured in venous blood samples from 38 to 72 Space Shuttle astronauts before and immediately after flights of 2 to 11 days. Mean pre- and postflight values were compared using the paired t-test or the Wilcoxon signed-rank test. The largest change in serum enzymes was a 21% increase (P = .0014) in gamma-glutamyl-transpeptidase, which may have been related to stress. The median value of apolipoprotein (apo) A-I decreased from 152 to 127 mg/dL (P < .0001), but the change in apo B (77 to 73 mg/dL) was not statistically significant, and the mean apo A-I/apo B ratio remained well above 1.5. A decrease in dietary fat and cholesterol intake during shuttle missions may have been a cause of the change in apo A-I. Twelve of the 16 nonenzyme serum proteins measured were significantly elevated (P < .05), possibly because of hemoconcentration and increased protein catabolism. The 56% increase in haptoglobin may be related to release of suppressed erythropoiesis at landing.

Influence of dietary calcium on chlordecone-induced biochemical changes in serum of rat.

Male, Sprague-Dawley rats were treated with different (1, 10, 50, and 100 ppm) concentrations of chlordecone (Cd) in calcium-sufficient (Ca-S) or calcium-deficient (Ca-D) diet for 15 days. No significant changes in serum total proteins were observed. However, serum nonprotein nitrogen compounds (urea, uric acid, and creatinine) and glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, creatine kinase, and alkaline phosphatase were significantly increased at 50 and 100 ppm of Cd. Chlordecone induced more increase in these serum components of rats fed with Ca-D as compared to Ca-S diet. Increased serum nonprotein nitrogen compounds and enzymes indicate Cd-altered glomerular and hepatic functions.