Ceruloplasmin Oxidase Activity Research Articles

Cerulenin upregulates heat shock protein-70 gene expression in chicken muscle

Lines of evidence suggested that systems involved in the regulation of the stress responses and energy homeostasis are highly integrated. Because cerulenin, the natural antibiotic product of the fungus Cephalosporium ceruleans and a broad-spectrum fatty acid synthesis (FAS) inhibitor, has been shown to affect food intake and energy balance, and because the biomarker of stress Hsp-70 gene was found to interact directly with fatty acids, we hypothesized that cerulenin may regulate Hsp-70 gene expression. Therefore, the present study was undertaken to examine this issue. Cerulenin administration significantly (P < 0.05) decreased food intake and induced Hsp-70 mRNA levels in muscle, but not in liver or hypothalamus of 2-wk-old broiler chickens. These changes were accompanied by an unpregulation of muscle uncoupling protein and carnitine palmitoyltransferase 1 mRNA levels. This result indicated that the regulation of Hsp-70 gene expression in normal chickens, as estimated by oxidative stress indices [TBA reacting substances, ferric reducing/antioxidant power, and ceruloplasmin oxidase activity] levels, is tissue-specific. In attempt to discriminate between the effect of cerulenin and cerulenin-reduced food intake on Hsp-70 gene expression, we also evaluated the effect of food deprivation on the same cellular responses. Food deprivation for 16 h did not affect Hsp-70 gene expression in all tissues examined, indicating that the effect of cerulenin is independent of the inhibition of food intake. To ascertain whether the effect of cerulenin is direct or indirect, we carried out in vitro studies. Cerulenin treatment did not affect Hsp-70 gene expression in Leghorn male hepatoma and quail myoblast cell lines, suggesting that the observed effect in vivo may be mediated through the central nervous system.

Oxytocin induced oxidative stress in lactating Bubalis bubalis (Nili Ravi)

BackgroundOxytocin has been reported for a wide range of adverse effects in different species of lactating animals. The present study was aimed to evaluate the adverse effects of oxytocin on biomarkers of oxidative stress in buffaloes. Lactating buffaloes (n = 40) were randomly selected from a commercial dairy farm located in the peri-urban area of district Faisalabad, Pakistan and divided into two equal groups viz. treatment and control groups, each containing 20 buffaloes. Buffaloes in treatment group were injected with oxytocin before each milking (morning and evening) for milk let down; whereas, animals in control group were milked naturally without oxytocin injection. Both the groups were assessed for oxidative stress biomarkers.ResultsResults showed significantly higher levels (P ≤ 0.05) of TOS, tHcy and ceruloplasmin oxidase activity in lactating buffaloes injected with oxytocin as compared to those of control group. On the other hand, serum levels of TAS, PON1 and arylesterase were significantly lower (P ≤ 0.05) in the buffaloes of treatment group.ConclusionsOxytocin injection in lactating buffaloes resulted in elevated oxidative stress by increasing the total homocysteine and ceruloplasmin oxidase activity and decreasing enzymatic activities of antioxidant enzymes including paraoxonase-1 and arylesterase; that might render the animals to poor productive and reproductive potential.

Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients

ENWEndNote BIBJabRef, Mendeley RISPapers, Reference Manager, RefWorks, Zotero AMA Cymerys M, Cofta S, Wysocka E, et al. Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients. Central European Journal of Immunology. 2013;38(4):511-517. doi:10.5114/ceji.2013.38249. APA Cymerys, M., Cofta, S., Wysocka, E., Nowicki, M., Suliburska, J., & Bryl, W. et al. (2013). Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients. Central European Journal of Immunology, 38(4), 511-517. https://doi.org/10.5114/ceji.2013.38249 Chicago Cymerys, Maciej, Szczepan Cofta, Ewa Beata Wysocka, Marcin Nowicki, Joanna Suliburska, Wiesław Bryl, and Lech Torliński. 2013. "Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients". Central European Journal of Immunology 38 (4): 511-517. doi:10.5114/ceji.2013.38249. Harvard Cymerys, M., Cofta, S., Wysocka, E., Nowicki, M., Suliburska, J., Bryl, W., and Torliński, L. (2013). Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients. Central European Journal of Immunology, 38(4), pp.511-517. https://doi.org/10.5114/ceji.2013.38249 MLA Cymerys, Maciej et al. "Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients." Central European Journal of Immunology, vol. 38, no. 4, 2013, pp. 511-517. doi:10.5114/ceji.2013.38249. Vancouver Cymerys M, Cofta S, Wysocka E, Nowicki M, Suliburska J, Bryl W et al. Oxidase activity of plasma ceruloplasmin in obstructive sleep apnea patients. Central European Journal of Immunology. 2013;38(4):511-517. doi:10.5114/ceji.2013.38249.

Mechanisims of asthma and allergic disease – 1066. Ceruloplasmin oxidase activity in allergic asthma and allergic rhinitis

Background The known functions of ceruloplasmin oxidase activity (COA) include copper transportation, iron metabolism, antioxidant defense and involvement in angiogenesis and coagulation. The role of ceruloplasmin oxidase involving the interaction of oxidant and anti-oxidant balance in allergic diseases is still unknown. It was previously reported that synthesis of ceruloplasmin was situmulated by interleukin1 in normal and copper-deficient rat models concluding that ceruloplasmin is dependent for oxidase activity. Moreover, the copper ions had been suggested as an explanation for the sensitivity of asthmatic individuals by their biologic effects of inhaled particulate air pollution. In vivo experiments on finding the cytokines involved in acute-phase protein response showed that there are three major cytokines; interleukin 1-beta, 6 and TNF-alpha. Our study was designed to examine the changes in COAs in severe persistent asthma-allergic rhinitis, new diagnosed allergic asthma-allergic rhinitis and allergic rhinitis patients.

Effects of iron injection at birth on neonatal iron status in young pigs from first-parity sows fed delta-aminolevulinic acid

A total of 40 gilts (Landrace×Yorkshire) fed delta-aminolevulinic acid (ALA) or inorganic iron (FeSO4) and their 392 progeny were used to determine the iron status of sows and piglets at birth and nursing. The experiment was a 2×2 factorial arrangement with two different additive [90mg of iron/kg (as FeSO4) and 90mg ALA/kg (ALA90)] and 2 levels of injection with iron-dextran (0 and 200mg) to neonatal pigs within litter. Dietary treatments were fed during entire gestation and lactation period. Sows fed ALA90 diet had larger (P<0.05) litter size born alive and piglets with higher (P<0.01) body weight at birth and weaning compared with sows fed FeSO4 diet. Plasma hemoglobin (Hb) and iron concentration at d 21, and milk iron concentration of sows were higher (P<0.05) when sows fed ALA90 vs. FeSO4. Feeding ALA90 to sows and iron injection to piglets increased body weight at d 0, 7 and 21 after birth and average daily gain from d 0–7. During d 0–7 the increase in average daily gain due to iron injection was higher in piglets of sows fed FeSO4 compared to sows fed ALA90 (ALA×iron injection; P<0.04). Feeding ALA90 to sows and iron injection increased (P<0.05) plasma iron and Hb concentration and hematocrit values of piglets throughout the nursing phase. At d 2 the increase in plasma iron and Hb concentration and hematocrit values of piglets due to iron injection was higher in piglets of sows fed FeSO4 compared sows fed ALA90 (ALA×iron injection; P<0.05). At d 7 and 21, plasma ceruloplasmin oxidase activity of piglets was reduced (P<0.05) by ALA supplementation to sows and iron injection. In conclusion, this study indicates that dietary supplementation of ALA to sows and iron injection to neonatal piglets can result in increased iron concentration in milk, which may have a beneficial effect on the plasma iron status of sucking pigs.

Systemic levels of ceruloplasmin oxidase activity in allergic asthma and allergic rhinitis

Context: The role of ceruloplasmin oxidase activity (COA) involving the interaction of oxidant and antioxidant balance in allergic diseases is still unknown.Objective: Our study was designed to examine the changes in COAs in severe persistent asthma-allergic rhinitis, new diagnosed allergic asthma-allergic rhinitis and allergic rhinitis patients.Methods: The study included 20 age- and sex-matched healthy individuals as control (Group I); Group II included 15 newly diagnosed allergic asthma-allergic rhinitis; Group III included 15 patients with severe persistent asthma-allergic rhinitis and in the fourth group there were 20 patients with allergic rhinitis. Group III was divided in two groups, severe persistent asthma-allergic rhinitis who were pre-(III-A) and post-treated (III-B) with omalizumab. Group IV was divided to two groups, pretreatment (IV-A) and posttreatment (IV-B) with specific subcutaneous immunotherapy modalities. All the posttreatment measurements were 12 months after the therapy. All the patients were assessed by the skin prick test, high sensitive C-reactive protein (hs-CRP) and COA.Results: There were significant differences between Group I and Groups III-A, III-B, IV-A and IV-B; Group II and Groups III-A, III-B, IV-A and IV-B; Group III-A and Groups III-B, IV-A and IV-B; Group III-A and Groups IV-A and IV-B; and Group IV-A and IV-B. Interestingly, there was a correlation between the hs-CRP and COA levels in Group III-A.Conclusions: Our data suggest that hs-CRP and COA levels might be an indicator of an inflammation and important in revelation of patients with allergy related diseases, especially of asthma patients.

Two-stage method for purification of ceruloplasmin based on its interaction with neomycin

A two-stage chromatography that yields highly purified ceruloplasmin (CP) from human plasma and from rat and rabbit serum is described. The isolation procedure is based on the interaction of CP with neomycin, and it provides a high yield of CP. Constants of inhibition by gentamycin, kanamycin, and neomycin of oxidase activity of CP in its reaction with p-phenylenediamine were assayed. The lowest K(i) for neomycin (11 µM) corresponded to the highest specific adsorption of CP on neomycin-agarose (10 mg CP/ml of resin). Isolation of CP from 1.4 liters of human plasma using ion-exchange chromatography on UNO-Sphere Q and affinity chromatography on neomycin-agarose yields 348 mg of CP with 412-fold purification degree. Human CP preparation obtained with A(610)/A(280) ~ 0.052 contained neither immunoreactive prothrombin nor active thrombin. Upon storage at 37°C under sterile conditions, the preparation remained stable for two months. Efficient preparation of highly purified CP from rat and rabbit sera treated according to a similar protocol suggests the suitability of our method for isolation of CP from plasma and serum of other animals. The yield of CP in three separate purifications was no less than 78%.

Repeated transplantation of hepatocytes prevents fulminant hepatitis in a rat model of Wilson's disease

The outcome of consecutive hepatocyte transplants was explored in a rat model of Wilson's disease before the onset of fulminant hepatitis without preconditioning regimens. Rats received a high-copper diet in order to induce a rapid induction of liver failure. Sham-operated rats (15/15) developed jaundice and fulminant hepatitis, and they died within 4 weeks of first transplantation. Despite the continuation of a high dietary copper challenge, long-term survival was observed for a notable proportion of the transplanted animals (7/18). All survivors displayed normalized levels of hepatitis-associated serum markers and ceruloplasmin oxidase activity by posttransplant days 50 and 98, respectively. The liver copper concentrations, the liver histology, and the expression of marker genes were significantly restored within 4 months of transplantation in comparison with the control group. The high expression of a copper transporter gene (ATPase Cu++ transporting beta polypeptide) in the livers of the survivors indicated a high rate of repopulation by donor hepatocytes. Our data suggest that repeated cell transplantation can overcome the limitations of a single therapy session in rats with severe hepatic disease by functionally restoring the host liver without preconditioning.

Early stage transplantation of bone marrow cells markedly ameliorates copper metabolism and restores liver function in a mouse model of Wilson disease

BackgroundRecent studies have demonstrated that normal bone marrow (BM) cells transplantation can correct liver injury in a mouse model of Wilson disease (WD). However, it still remains unknown when BM cells transplantation should be administered. The aim of this study was to investigate the potential impact of normal BM cells transplantation at different stages of WD to correct liver injury in toxic milk (tx) mice.MethodsRecipient tx mice were sublethally irradiated (5 Gy) prior to transplantation. The congenic wild-type (DL) BM cells labeled with CM-DiI were transplanted via caudal vein injection into tx mice at the early (2 months of age) or late stage (5 months of age) of WD. The same volume of saline or tx BM cells were injected as controls. The DL donor cell population, copper concentration, serum ceruloplasmin oxidase activity and aspartate aminotransferase (AST) levels in the various groups were evaluated at 1, 4, 8 and 12 weeks post-transplant, respectively.ResultsThe DL BM cells population was observed from 1 to 12 weeks and peaked by the 4th week in the recipient liver after transplantation. DL BM cells transplantation during the early stage significantly corrected copper accumulation, AST across the observed time points and serum ceruloplasmin oxidase activity through 8 to 12 weeks in tx mice compared with those treated with saline or tx BM cells (all P < 0.05). In contrast, BM cells transplantation during the late stage only corrected AST levels from 4 to 12 weeks post-transplant and copper accumulation at 12 weeks post-transplant (all P < 0.05). No significant difference was found between the saline and tx BM cells transplantation groups across the observed time points (P > 0.05).ConclusionsEarly stage transplantation of normal BM cells is better than late stage transplantation in correcting liver function and copper metabolism in a mouse model of WD.

A preliminary investigation on the possible association between diminished copper availability and non-alcoholic fatty liver disease in epileptic patients treated with valproic acid

Background. Patients treated with valproic acid (VPA) present a high incidence of non-alcoholic fatty liver disease (NAFLD) (around 61%). Several recent studies suggest that low copper stores could be associated with NAFLD, and a significant decrease of copper availability in VPA-treated patients has been described.Design and methods. In 101 adult epileptic patients treated with valproic acid in monotherapy (n = 75) and polytherapy (n = 26) the copper availability was evaluated using the specific oxidase activity of ceruloplasmin (activity per unit mass of enzyme protein) and the copper/ceruloplasmin ratio. Copper deficiency was supposed in the cases in which this biochemical variable was smaller than the lower reference limit (333 U/g).Results. The differences between the groups of patients with ceruloplasmin oxidase activity smaller or greater than 333 U/g for the serum levels of aminotransferases, gamma-glutamyltransferase, butyrylcholinesterase, cholesterol, triglycerides, and C-reactive protein, and the APRI and FIB-4 liver fibrosis scores were not statistically significant. Most patients (93%) had low APRI and FIB-4 scores, suggesting absence of significant liver fibrosis.Conclusions. The results obtained do not confirm the hypothesis of an association between diminished copper availability and NAFLD in patients treated with valproic acid.

Study of Interaction of Ceruloplasmin with Serprocidins

This paper describes formation of complexes of ceruloplasmin (CP) with such proteins of the serprocidin family as azurocidin (CAP37), neutrophilic elastase (NE), cathepsin G (CG), and proteinase 3 (PR3). We present evidence that serprocidins form complexes with CP at a molar ratio 1 : 1. Phenylmethylsulfonyl fluoride, a serine protease inhibitor, did not prevent the interaction of serprocidins with CP in the course of SDS-free disc electrophoresis. CP affected the activities of NE, CG, and PR3 as a competitive inhibitor with K(i) ~ 1 μM. Inhibitory effect of CP depended on ionic strength of the solution and was negligible at NaCl concentrations above 300 mM. In the mode of competitive inhibitors serprocidins suppressed oxidase activity of CP towards p-phenylenediamine. CAP37 displayed the strongest inhibitory effect (K(i) ~ 20 nM). Upon adding various serprocidins to human, rat, rabbit, dolphin, dog, horse, and mouse plasma only CAP37 would form a complex with CP. Synthetic peptide RKARPRQFPRRR (5-13, 61-63 CAP37) displaced CAP37 from its complex with CP. Adding CAP37 to the triple complex formed by CP, lactoferrin, and myeloperoxidase resulted in displacement of the latter from the complex. The dissociation constant of CAP37 with immobilized CP was 13 nM. Therefore, among serprocidins CAP37 can be regarded as the specific partner of CP.

Effect of valproic acid treatment on copper availability in adult epileptic patients

Objectives In rats a significant increase of the copper biliary excretion is produced by valproic acid administration, however, a conclusive study on the possible appearance of copper deficiency in humans during treatment with this drug has still not been carried out. Design and methods In 101 adult epileptic patients treated in monotherapy ( n = 75) and polytherapy ( n = 26) with valproic acid, and 50 healthy controls, were determined serum copper, immunoreactive ceruloplasmin and its oxidase activity against o-dianisidine, in order to calculate the specific oxidase activity (activity per unit mass of enzyme protein) and copper/ceruloplasmin ratio. Results Specific oxidase activity of ceruloplasmin and copper/ceruloplasmin ratio were significantly lower in the groups of patients treated with valproic acid than in the controls. Significant correlations were obtained between both biochemical variables ( p < 0001). In 38% of the patients treated in mono or polytherapy, the specific oxidase activities of ceruloplasmin were smaller than the estimated lower limit of reference. Conclusions These results, possible due to a diminished copper content of the circulating ceruloplasmin, suggest that marginal or moderate copper deficiency may have a substantial prevalence among patients treated with valproic acid.

Evidence for a critical role of ceruloplasmin oxidase activity in iron metabolism of Wilson disease gene knockout mice

Wilson disease is a genetic disorder associated with copper overload due to mutations within the ATP7B gene. Although copper and iron metabolism are closely linked, the influence of mutations of the ATP7B gene on iron homeostasis is unknown. Therefore, the present study was carried out to elucidate iron metabolism in Atp7b(-/-) mice, an animal model of Wilson disease. Hepatic iron content, serum iron parameters and blood hemoglobin levels of Atp7b(-/-) mice and wild type mice were studied. Hepatic and duodenal expression of iron metabolism-related genes was measured quantitatively by real-time reverse transcription-polymerase chain reaction and post-translational expression of Dmt1 was analyzed by immunoblot. Atp7b(-/-) mice displayed copper accumulation (P < 0.001), slightly elevated hepatic iron content (P = NS), and a low serum ceruloplasmin oxidase activity (1.5 +/- 1.9 U/L vs 18.9 +/- 4.0 U/L, P < 0.001) when compared with wild type mice. Serum iron, serum transferrin saturation, and blood hemoglobin levels were significantly lower in Atp7b(-/-) mice compared with controls (121.2 +/- 35.3 microg/dL vs 201.8 +/- 34.9 microg/dL (P < 0.001); 44.0 +/- 12.7% vs 68.0 +/- 8.2% (P < 0.001); and 12.7 +/- 0.2 g/dl vs 15.3 +/- 0.1 g/dl (P < 0.001), respectively). Hepatic mRNA expression of hepcidin, TfR-1, TfR-2, hemojuvelin, and Dmt1 + IRE did not differ significantly between Atp7b(-/-) and wild type mice. In the duodenum of Atp7b(-/-) mice Dmt1 + IRE and hephaestin did not show any differences in their mRNA levels compared with wild type mice, while Dcytb mRNA expression was 1.7-fold increased compared with wild type mice (P = 0.01). Atp7b(-/-) mice demonstrated decreased serum iron parameters and hemoglobin levels most likely related to a low serum ceruloplasmin oxidase activity and not due to total body iron deficiency.

Truncating mutations in the Wilson disease gene ATP7B are associated with very low serum ceruloplasmin oxidase activity and an early onset of Wilson disease

BackgroundMutations in the gene ATP7B cause Wilson disease, a copper storage disorder with a high phenotypic and genetic heterogeneity. We aimed to evaluate whether 'severe' protein-truncating ATP7B mutations (SMs) are associated with low serum ceruloplasmin oxidase activities and an early age of onset when compared to missense mutations (MMs).MethodsThe clinical phenotype of 59 genetically confirmed WD patients was analyzed retrospectively. Serum ceruloplasmin was measured by its oxidase activity with o-dianisidine dihydrochloride as substrate and immunologically.ResultsThirty-nine patients had two MMs, 15 had the genotype SM/MM, and 5 patients had two SMs on their ATP7B alleles. Enzymatic and immunologic serum ceruloplasmin levels differed significantly between the three groups (P < 0.001 and P < 0.01, respectively). The lowest levels were measured in patients with two SMs (0.0 U/L; IQR, 0.0-0.0 U/L and 0.02 g/L; IQR, 0.01-0.02 g/L, respectively) and the highest in patients with two MMs (17.8 U/L; IQR, 5.8-35.1 U/L and 0.11 g/L; IQR,0.10-0.17 g/L, respectively). The age of onset was also significantly different between the three patient groups (P < 0.05), with SM/SM patients showing the earliest onset (13 years; IQR, 9-13 years) and patients with two MMs showing the latest onset (22 years; IQR, 14-27 years). By ROC curve analysis a ceruloplasmin oxidase level ≤ 5 U/L can predict the presence of at least one SM with a sensitivity of 80% and a specificity of 79.5%.ConclusionsIn our German study cohort truncating ATP7B mutations were associated with lower ceruloplasmin serum oxidase levels and an earlier age of onset when compared to MMs. Measurement of serum ceruloplasmin oxidase might help to predict presence of truncating ATP7B mutations and might facilitate the mutation analysis.

Deneysel Abdominal Kompartman Sendromu Oluşturulan Sıçanlarda Serum Malondialdehid Düzeyi ile Seruloplazmin Oksidaz Aktivitesi

Objective: The aim of this study was to investigate serum ceruloplasmin (Cp) oxidase activity and malondialdehyde (MDA) levels in a rat model of abdominal compartment syndrome (ACS). Material and Methods: There were four groups consisting of eight rats per group. Group 1 (Control group): After anesthesia, 3 ml of blood were taken from the heart of these rats. Group 2 (Sham group): Under anesthesia, an injector needle was inserted intraperitoneally and left there. After 3 h, 3 ml of blood was taken from the heart of these rats. Group 3 (First study group): Under anesthesia, an injector needle (number 16) was inserted intraperitoneally and a standard insufflator (Karl Storz, Germany) was connected. At the end of this procedure, the abdomen was decompressed and 3 ml of blood was collected as descried above. Group 4 (Second study group): The abdomen was compressed as Group 3. However, the intraabdominal hypertension was established by keeping the pressure constant at 25 mmHg for 3 h in this group. At the end of this procedure, the abdomen was decompressed and 3 ml of blood was collected as descried above. Results: Cp oxidase activities and MDA levels significantly increased in first and second study groups when compared to the control group. Cp oxidase activities and MDA levels were significantly higher in second study group than those of the first study group. MDA levels were significantly higher in first study group than those of the sham group. Conclusion: We found increased serum Cp oxidase activity and MDA level in experimental abdominal compartment syndrome-induced rats. These findings show that oxidative stress increase ACS in rats.

Antioxidant potential, paraoxonase 1, ceruloplasmin activity and C-reactive protein concentration in diabetic retinopathy

The aim of this study was to evaluate the ferric-reducing ability of serum (FRAS), paraoxonase 1 (PON1), ceruloplasmin serum oxidase activity and hsCRP level in patients with type1 diabetes mellitus without and with diabetic retinopathy. The study was performed in 76 patients with type 1 diabetes mellitus, 35 without diabetic retinopathy (group 1) and 41 with preproliferative and proliferative retinopathy (group 2). Control group consisted of 35 nondiabetic, age-, gender-, body mass-matched healthy volunteers who came to the outpatient clinic for a routine health check-up. We evaluated FRAS using the method described by Benzie and Strain; PON1 by kinetic spectrophotometric assay with paraoxon as substrate and ceruloplasmin using its oxidative activity with 3-phenylenodiamine as substrate. CRP was measured with a high sensitive enzyme immunoassay. PON1 activity was significantly decreased in patients with diabetic retinopathy (227.66 +/- 123.57 U/l) when compared with control (312.04 +/- 129.77 U/l). FRAS was significantly decreased in group 2 (439.33 +/- 79.87 micromol/l) when compared with group 1 (522.79 +/- 167.56 micromol/l) and control (529.80 +/- 81.99 micromol/l). Ceruloplasmin activity was significantly elevated in group 1 (58.36 +/- 22.56 U/g protein) when compared with control (45.22 +/- 14.96 U/g protein). We have found significant increase in hsCRP level in group 2 (3.71 +/- 2.47 mg/l) when compared with group 1 (1.75 +/- 1.01 mg/l) and control (0.57 +/- 0.46 mg/l). The PON1/CRP ratio in control group was significantly increased when compared with diabetic patients and was significantly decreased in group 2 compared with group 1. We have not found gender-dependent difference in studied parameters in both control and in study groups. We have found tendency to decrease the serum activity of FRAS and hsCRP in elder patients but the difference was significant only in group 2. FRAS and PON 1 activity is decreased in patients with type 1 diabetes mellitus with presence of diabetic retinopathy which confirms that oxidative stress could play a role in pathogenesis of diabetic retinopathy. Significantly elevated levels of hsCRP in diabetic patients with the presence of diabetic retinopathy compared with patients without diabetic retinopathy providing a link between inflammation and the development of microvascular complication of diabetes. Because of the significant difference in PON1/CRP ratio between patients without and with the presence of diabetic retinopathy, it seems that PON1:CRP ratio may be used as a biochemical marker for progression of retinopathy. The link between the antioxidant concentration, inflammation and the development of diabetes complications needs further longitudinal studies in order to confirm our findings.

Serum ceruloplasmin oxidase activity is a sensitive and highly specific diagnostic marker for Wilson’s disease

A low serum ceruloplasmin concentration is considered diagnostic for Wilson's disease. We aimed to evaluate an enzymatic test for ceruloplasmin oxidase activity and to compare it with the routinely used immunological ceruloplasmin measurement. Serum ceruloplasmin was measured enzymatically with o-dianisidine dihydrochloride as substrate and immunologically. 110 Wilson's disease patients, 52 healthy controls, and 51 patients with impaired liver function not due to Wilson's disease were analyzed. Assay performance was tested by receiver operating characteristic curve analysis, McNemar test, and Spearman's rank correlation. The greatest sum of sensitivity and specificity was seen for the enzymatic ceruloplasmin assay at a cut-off point of 55 U/L (93.6% and 100%, respectively) and for the immunologic assay at a cut-off point of 0.19 g/L (93.6% and 78.8%, respectively). For healthy controls, the differences in specificity between both assays were statistically significant (McNemar, p=0.02). When additionally including patients with impaired liver function into the control group the specificity declined to 84.5% for the enzymatic assay and to 68.9% for the immunologic assay. The correlation between the enzymatic and immunologic assay was high in healthy controls (r=0.94), but weaker in Wilson's disease patients (r=0.70) and patients with impaired liver function not due to Wilson's disease (r=0.65). For the enzymatic assay the best cut-off point for predicting Wilson's disease was estimated to be 55 U/L. Our data suggest that the enzymatic ceruloplasmin assay is superior to the immunologic assay in diagnosing Wilson's disease and should become the preferred method.

Antioxidants in Colostrum and Milk of Sows and Cows

On account of the oxidative stress conditions that may appear during parturition, colostrum should provide with not only nutritional and immunological components but also antioxidative protection of newborn. There is evidence that apart from well-known antioxidative enzymes like glutathione peroxidase, superoxide dismutase, catalase or low molecular antioxidants, proteins like lactoperoxidase (LPO), lactoferrin (LF) and ceruloplasmin (CP) may exert antioxidative properties in colostrum. The aim of present study was to determine and to evaluate LPO, LF and CP activities in colostrum and milk of sows and cows. Samples were collected from 16 healthy cows five times: immediately after parturition, 12, 24 and 48 h, and 7 days postpartum as well as from 14 healthy sows five times: immediately after parturition, 6, 12, 24 and 36 h later. Examined parameters were determined spectrophotometrically at 412, 560 and 540 nm respectively. LPO activity was higher in sows as in cows and increased significantly within examined time. LF ability to inhibit superoxide radical generation was higher in sows as in cows and increased significantly within examined time. CP oxidase activity was higher in cows as in sows and decreased significantly during experimental period. In conclusion, antioxidative defence system in colostrum shows dynamic changes that allow for providing with necessary protection from oxidative stress conditions, which may appear after parturition.

Melatonin improves oxidative stress parameters measured in the blood of elderly type 2 diabetic patients

An elevated oxidative status in the aging organism may be involved in the development of non-insulin dependent diabetes mellitus (NIDDM). Melatonin, a potent antioxidant agent, is essential for glucose homeostasis and regulation. The aim of this study was to determine the influence of melatonin supplementation on the oxidative stress parameters in elderly NIDDM patients. The malondialdehyde (MDA) concentration, Cu-Zn superoxide dismutase (SOD-1) activity in erythrocytes, the level of nitrate/nitrite in plasma and morning melatonin concentration and oxidase activity of ceruloplasmin (Cp) in serum in 15 elderly NIDDM patients at baseline and after the 30 days of melatonin supplementation (5 mg daily) in comparison with levels in 15 healthy elderly volunteers were determined. A significant increase of MDA level and decrease of SOD-1 activity and melatonin concentration were observed in NIDDM patients. Cp oxidase activity and nitrate/nitrite level were similar in both examined groups. Melatonin administration in NIDDM patients resulted in a significant increase in the morning melatonin concentration and SOD-1 activity, and a reduction in the MDA level and Cp oxidase activity. Statistically significant alterations in nitrate/nitrite levels were not observed. These results indicate an improvement of antioxidative defense after melatonin supplementation in the NIDDM individuals and suggest melatonin supplementation as an additional treatment for the control of diabetic complications.

Effects of neonatal iron status, iron injections at birth, and weaning in young pigs from sows fed either organic or inorganic trace minerals1,2,3

Second-parity sows (n = 7) were fed diets containing organic or inorganic trace minerals, and their progeny (n = 68) were used to determine the Fe status of pigs at birth and nursing and postweaning phases. The experiment comprised 2 parts, in which the first experiment was a 2 x 2 factorial arrangement. Sow trace mineral (organic vs. inorganic) was the first factor evaluated, and the injection of Fe (0 or 200 mg) to neonatal pigs within litter was the second factor. In Exp. 2, half the pigs in each litter from each neonatal Fe injection group were injected with Fe (0 vs. 200 mg) at weaning as an added factor in a 2 x 2 x 2 factorial arrangement in a split-split-plot design. Weanling pigs were fed diets fortified with 90 mg/kg of Fe (sulfate), but the analyzed indigenous and fortified Fe content was 170 mg/kg. Pigs in both experiments were bled at periodic intervals to determine hemoglobin (Hb) concentration, percentage of hematocrit (Hct), and ceruloplasmin oxidase activity. Neonates and d-2 pigs from sows fed organic trace minerals had lower (P < 0.05) Hb concentrations compared with sows fed inorganic trace minerals, but they had similar percentages of Hct values. Blood Hb seemed to remain lower throughout the nursing period when sows were fed organic vs. inorganic Fe. Pigs without Fe injection had decreased ADG (P < 0.05) from 0 to 7 and 7 to 17 d than pigs injected with Fe. Although Hb values increased when neonatal pigs received Fe injection, they were somewhat lower when sows were fed the organic Fe. Ceruloplasmin oxidase activity was low at birth, increased to weaning in each treatment group, and was greater in pigs without Fe injection at d 13 (P < 0.05) and those from sows fed organic minerals at d 17 (P < 0.01). In Exp. 2, when the Fe-fortified diet was fed, BW and ADG responses were both greater (P < 0.01) to 28 d postweaning when neonates had received Fe injections. Neonates not injected with Fe at birth but injected at weaning had greater ADG, Hb, and Hct values, whereas pigs injected with Fe did not respond to Fe injection at weaning, which resulted in interactions (P < 0.05) in those criteria at most measurement periods. The results indicated a reduced Fe bioavailability when sows were fed the organic Fe source, but this may also have been due to the greater Fe need, lowered Fe status, or both, of the sow because of the greater number of pigs farrowed and heavier litter weights at parturition and weaning. The results also indicated that Fe injections at birth may be critical to achieving maximum pig growth response to weaning. There was no apparent advantage to injecting Fe at weaning when neonatal pigs received Fe injections.