Ceruloplasmin replacement therapy ameliorates neurological symptoms in a preclinical model of aceruloplasminemia.

Aceruloplasminemia is a monogenic disease caused by mutations in the ceruloplasmin gene that result in loss of protein ferroxidase activity. Ceruloplasmin plays a role in iron homeostasis, and its activity impairment leads to iron accumulation in liver, pancreas, and brain. Iron deposition promotes diabetes, retinal degeneration, and progressive neurodegeneration. Current therapies mainly based on iron chelation, partially control systemic iron deposition but are ineffective on neurodegeneration. We investigated the potential of ceruloplasmin replacement therapy in reducing the neurological pathology in the ceruloplasmin-knockout (CpKO) mouse model of aceruloplasminemia. CpKO mice were intraperitoneal administered for 2 months with human ceruloplasmin that was able to enter the brain inducing replacement of the protein levels and rescue of ferroxidase activity. Ceruloplasmin-treated mice showed amelioration of motor incoordination that was associated with diminished loss of Purkinje neurons and reduced brain iron deposition, in particular in the choroid plexus. Computational analysis showed that ceruloplasmin-treated CpKO mice share a similar pattern with wild-type animals, highlighting the efficacy of the therapy. These data suggest that enzyme replacement therapy may be a promising strategy for the treatment of aceruloplasminemia.

[Efficacy of ceruloplasmin in patients with asthma].

AIM: To evaluate the efficacy ceruloplasmin (Cp) used in the combination therapy of patients with an asthma exacerbation. SUBJECTS AND METHODS: The trial included 37 asthmatic patients. Chemiluminescence (ChL) registration was used to study the production of reactive oxygen species (ROS) in the patients' blood. Nineteen patients with asthma received conventional treatment. Cp was used as part of combination therapy in 18 asthmatic patients. RESULTS: ChL intensity was increased the blood of patients with an asthma exacerbation. Cp treatment resulted in a reduction in the generation of ROS in the blood and contributed to positive changes in the clinical symptoms of asthma. In the patients receiving conventional therapy, the high ChL intensity of blood was retained and the clinical symptoms of the disease reduced. CONCLUSION: The use of Cp in patients with an asthma exacerbation corrects free radical oxidation processes and enhances therapeutic effectiveness.

[Dynamics of oxidase activity in blood plasma upon intravenous and intramuscular administration of ceruloplasmin]. / Dinamika izmeneniia oksidaznoi aktivnosti plazmy pri razlichnikh sposobakh vvedeniia preparata Tseruplazmin.

Oxidase activity dynamics in the blood plasma of test animals was studied upon intravenous and intramuscular injections of ceruloplasmin. It was found that the enzymatic activity does not reflect the content of drug in the blood plasma and is not always correlated with the amount of exogenously introduced drug. Intramuscular injections can also be used for the administration of ceruloplasmin.

[A comparative study of the transport dynamics of the peptide moiety of the milk ceruloplasmin molecule in the body of rats with embryonic and adult types of copper metabolism]. / Sravnitel'noe izuchenie dinamiki peremeshcheniia peptidnoi chasti molekuly tseruloplazmina moloka v organizme krys pri émbrional'nom i vzroslom tipakh metabolizma medi.

In chase experiments, we followed the distribution of [125I]-ceruloplasmin prepared from human breast milk orally administered to young rats. Experiments were conducted using six-day-old rat pups (the embryonic type of copper metabolism) or 35-day-old ones (the adult type of copper metabolism). Using the technique of rocket immunoelectrophoresis, we have demonstrated that in six-day-old rats [125I]-ceruloplasmin was transferred from the gastrointestinal tract to the bloodstream and could be detected there over a period of 4 h. In 35-day-old rats, milk ceruloplasmin was digested in the upper part of the intestinal tract. The dynamic aspects of the distribution of labeled milk ceruloplasmin in the body of six-day old rats over a period of 4 h point out that, under the conditions of embryonic copper metabolism, it can serve as a transporter of copper ions to extrahepatic organs. We discuss the role of milk ceruloplasmin in copper metabolism in mammals during the neonatal period.

Synthesis and turnover of ceruloplasmin in rats treated with 17 beta-estradiol.

The effects of estrogen on synthesis and turnover of ceruloplasmin were studied in adult female Fischer rats. Daily treatment with 140 microgram 17 beta-estradiol resulted in a slow rise of ceruloplasmin concentrations, as measured by p-phenylenediamine oxidase activity, leading to a 70% increase by 7 days and a tripling by Day 14. Ceruloplasmin protein concentrations increased to the same degree, based on yields of the protein obtained during purification. Effects of estrogen on rates of synthesis (incorporation of [3H]leucine) were followed, using immunoprecipitation of total ceruloplasmin or isolation of its two major isoforms (Rfs 0.4 and 0.6 in native gel electrophoresis). Synthesis was increased by 7 days and was 2.5 times that of controls by Day 14. Both forms of ceruloplasmin showed the same specific activities and degree of increase in rate of synthesis. Rates of ceruloplasmin turnover were unaffected, based on double labeling with 3H- and 14C-leucine, but were three- to fourfold faster than for total plasma protein. The enzymatically more active 0.6 Rf form of ceruloplasmin had a faster turnover rate than the 0.4 Rf form. Estrogen treatment doubled ceruloplasmin mRNA levels by 7 days and almost tripled them by Day 14. Most of the ceruloplasmin mRNA was associated with the endoplasmic reticulum-bound polyribosomes. We conclude that estrogen increases the rate of synthesis of two forms of ceruloplasmin by indirectly increasing liver concentrations of its mRNA but has no effect on ceruloplasmin turnover.

[Determination of oxidative activity of ceruloplasmin using Ravin's method in patients with laryngeal carcinoma]. / Ocena aktywnosci oksydazowej ceruloplazminy oznaczanej metoda ravina u chorych na raka krtani.

According to Ravin's method an oxidative activity of ceruloplasmin was determined in the group of patients with carcinoma of the larynx before and after a surgical treatment, in the group of patients with inflammatory lesions in the upper respiratory tract and in the control group of healthy individuals. As a result of this assay the significantly increased of oxidative activity ceruloplasmin was obtained in the individuals with the carcinoma of the larynx (before surgical treatment) in comparison to the group of patients suffering due to inflammatory diseases of the upper respiratory tract and to the group of healthy subjects. Simultaneously a significant decrease of the oxidative activity of serum ceruloplasmin in the individuals with carcinoma of the larynx after a previous surgical treatment was observed.

Interaction of 125I-labelled ceruloplasmin with human trophoblast cells in vitro.

In vitro binding, internalization and release of the plasma protein ceruloplasmin was investigated using primary cultures of human placental trophoblast cells. Binding of 125I-labelled ceruloplasmin at 4 degrees C reached equilibrium by 5-6 h; binding was linear throughout all concentrations tested (1 nM-3.3 microM). Addition of greater than 5-10 microM unlabelled ceruloplasmin or a variety of other proteins (albumin, transferrin, IgG) were equally effective in displacing bound ceruloplasmin in a concentration-dependent manner. When cells were incubated at 37 degrees C, the majority of surface-bound 125I-labelled ceruloplasmin was released directly to the extracellular medium. Trypsin-resistant radioactivity increased to 18 per cent of initially bound ceruloplasmin within 1 min, declining to 5 per cent by 2 h. The acquisition of trypsin-resistant radioactivity was unaffected by the addition of a variety of metabolic inhibitors and no evidence of intracellular degradation of ceruloplasmin was found. In summary, our results suggest that the majority of ceruloplasmin binding to trophoblast cells is nonspecific, of low affinity, and easily dissociable at 4 degrees C. Only a small amount of ceruloplasmin appeared to be internalized, by an as yet unknown mechanism.

[Effect of selected physico-chemical parameters on changes in ascorbate oxidase and ceruloplasmin activities]. / Wplyw wybranych parametrów fizykochemicznych na zmiany aktywnosci oksydazy askorbinianowej i ceruloplazminy.

The paper presents the results of kinetic studies covering the reactions with the participation of two enzymes belonging to the group of "blue oxidases"--ascorbate oxidase and ceruloplasmin. Using variable physico-chemical parameters of reactions such as: temperature, presence of denaturizing factors, various substrates, it was possible to draw conclusions as to the structure and mechanism of reactions catalyzed by these enzymes. The following findings were established: 1. Ceruloplasmin is characterized by the absence of quarternary structure and lesser substrate specificity as compared with ascorbate oxidase. 2. The mechanism of reactions catalyzed by ceruloplasmin and ascorbate oxidase is different, probably due to the fact that there is no typical active centre binding the substrate in ceruloplasmin. 3. The application of variable parameters of physico-chemical reactions in the kinetic studies facilitates the description of the structures of enzymes and the mechanism of reactions being catalyzed by them.

Talidomida efeito sobre a ceruloplasmina serica / Thalidomide: effect on the serum ceruloplasmin

Ceruloplasmin enzymatic activity was determined in rat sera using p-phenylenediamine as substrate at pH 5.4. The levels of the oxidase in the animais treated with thalidomide by perenteral or gatro-intestinal routes were not statistically different from the values obtained in the control rats.