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.

[Clinical manifestations of asthma during combination therapy using ceruloplasmin].

AIM: To estimate the time course of clinical changes in patients with asthma during combination therapy using ceruloplasmin (CP). MATERIALS AND METHODS: A total of 92 asthmatic patients were examined. Their medical history data were collected; external lung function testing and clinical, laboratory, and instrumental examinations, involving the determination of the indicators of lipid peroxidation (LPO) (malonic dialdehyde (MDA), methemoglobin, carboxyhemoglobin) and the antioxidant system (superoxide dismutase (COD), sulfhydryl groups), were performed in all the patients over time. According to the therapy used, the patients were divided into 2 groups matched for gender, age, and clinical manifestations of the disease. A study group consisted of 45 patients who took CP in addition to conventional therapy. A comparison group included 47 patients receiving standard therapy. RESULTS: During the combination therapy using CP, the asthmatic patients showed a reduction in the elevated concentrations of MDA, methemoglobin, and carboxyhemoglobin and increases in the activity of COD and in the levels of sulfhydryl groups, which was followed by a considerable clinical improvement. During the conventional therapy, the indicators of LPO remained high and those of the antioxidant system did low, suggesting permanent oxidative stress. CONCLUSION: CP incorporation into the combination therapy of asthmatic patients contributes to elimination of prooxidant-antioxidant imbalance, which is followed by a marked positive clinical effect.

Determinação de cobre e outros minerais em caprinos e ovinos criados no sertão do vale do rio São Francisco, Pernambuco / Determination of copper and other minerals in goats and sheep of the San Francisco river valley in Pernambuco, Brazil

Para a determinação dos teores de cobre e de seus antagonistas, foram utilizadas 160 amostras de soro e de fígados, de caprinos e ovinos enviados ao matadouro municipal de Petrolina. As amostras de fígado e soro foram correlacionadas para o mesmo animal, a fim de evitar erros na obtenção dos dados. No soro a atividade da ceruloplasmina foi determinada por método colorimétrico. Para a determinação dos minerais, as amostras foram diluídas de seis a vinte vezes com água Milli-Q. Para determinação das concentrações dos elementos minerais no fígado, as amostras foram digeridas até que se obtivesse uma solução que mantivesse os minerais da amostra inicial e que fosse totalmente liquida, sem a presença de partículas sólidas que pudessem obstruir os capilares de sucção do espectrômetro e assim impedir as leituras das amostras. As concentrações de cobre, molibdênio, ferro e zinco foram determinadas através de espectrometria óptica por emissão de plasma (ICP). Desta forma, foi conduzido o experimento objetivando determinar a ocorrência e distribuição da carência de cobre no território do sertão do vale do rio São Francisco em Pernambuco. Foi observado que não houve carência de cobre nesta região do estado de Pernambuco, quando se avaliou os níveis médios de cobre hepático,. Os níveis de zinco estavam dentro de um padrão de normalidade, enquanto que os níveis de ferro foram mais elevados em ovinos, e os níveis de molibdênio mais reduzidos em caprinos. Verificou-se também que a atividade de ceruloplasmina foi um indicador dos níveis séricos de cobre...

For the determination of copper concentration and its antagonists, 160 serum and liver samples were used, from goat and sheep sent to the municipal slaughterhouse of Petrolina. The samples were correlated with the same animal, in order to prevent errors in data collection. Serum ceruloplasmin activity was determined by colorimetric method. For determination of minerals, the samples were diluted six to twenty times with Milli-Q water. For determining concentrations of mineral elements in the liver, the samples were digested until achieving a solution that maintained the initial sample and minerals totally liquid, without the presence of solid particles which could clog the capillary suction of the spectrometer and prevent reading of samples. The concentrations of copper, molybdenum, iron and zinc were determined by optical emission spectrometry (ICP). Thus, the experiment was conducted to determine the occurrence and distribution of copper deficiency in the San Francisco valley of Pernambuco. No copper deficiency was revealed in this region of the state of Pernambuco, when the mean levels of hepatic copper were evaluated. Zinc levels were within a normal range, whereas iron levels were higher in sheep, and lower levels of molybdenum in goats. It was also found that the activity of ceruloplasmin was an indicator of serum copper...

[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.

[Chronic pyelonephritis in workers of chemical industry: pathogenesis and treatment].

The examination of 500 workers engaged in the production of chemicals detected chronic pyelonephritis in 81 (16%) of them. Such occupational hazards as furnace-charge dust, fly-powder, calcium petre in the air, intensify their toxic action on workers in high temperature and humidity (54-67%) at workplace and provoke pyelonephritis. An experimental model of pyelonephritis was developed in animals exposed to the above environmental hazards. Ceruloplasmin added to combined treatment of chronic pyelonephritis relieves endotoxicosis by reducing content of middle-mass molecules in blood plasma.

[Combination of antioxidants and a hepatoprotector in the treatment of patients with diabetic distal symmetric polyneuropathy].

The complex open randomized clinical, instrumental and biochemical research with including 89 patients with type 2 diabetes mellitus (DM), complicated with distal symmetric polyneuropathy (DDSP) has been conducted. It is proved that the antioxidant "cocktail" application (Dialipon, Byocerulin, hepatoprotector with the antioxidant action Glutargin) contributes to the improvement of different levels of antioxidant defence, reduces free radical processes intensification, improves hepatocyte state which dysfunction is one of leading links of DDSP pathogenesis. These improves indirectly and directly the peripheral nerves functional state according electroneuromyography results, prevents DDSP progression and development of diabetic foot syndrome. Taking into account negative influence of Dialipon on the clinical parameters of DDSP with parallel intensification of free radical processes at decompensated DM in the early terms of treatment, Dialipon use is recommended after normalization of glycemia. At the same time the positive influence of Byocerulin has been noted regardless the compensation state of diabetes.

[The influence of ceruloplasmin on the rat neocortical ultrastructure in circulatory hypoxia].

Diffuse cerebral ischemia and circulatory hypoxia were modeled by carotid arteries occlusion in 40 white Wistar male rats. The changes of microcirculation bed and neurons of the sensomotor cortical area in hypoxia, preventive and therapeutic regimes of ceruloplasmin injections were compared to the neocortex state of intact rats. The changes of the neocortical ultrastructure established by electron microscopy were characteristic of hypoxic lesions described earlier in the literature. As a result of prolonged administration of ceruloplasmin there was a significant decrease of sensomotor neocortical hypostasis and improvement of microcirculation that suggested its neuroprotective properties.

[Milk ceruloplasmin as a physiological source of nutritional copper in early ontogenesis in mammals].

This work elucidates the role of ceruloplasmin (Cp): the main soluble copper containing glycoprotein, in newborn infant copper metabolism. An over 3-fold drop of Cp and copper concentration in samples of skimmed milk from 2 to 5 days of lactation was demonstrated. It has been shown that [125I]Cp of the breast milk is discovered in the blood plasma after its was administered per os to 6-day old rats. In the body, [125I]Cp was specifically bound to cells and lost copper ions. Alternatively, after copper metabolism change to adult type, milk [125I]Cp isn't carried from gastrointestinal tract to the blood. During 8 days, newborn rats were fed with baby formula and we have found that in this case changing of the copper metabolism type takes place earlier in contrast to the litter fed by females. Transcription of the Cp gene activated in liver, Cp and the copper ions concentration in the blood plasma increased proportionally, whereas in liver the copper concentration is decreased. In the brain, changes typical for adult copper metabolism, were not discovered. But in cerebrospinal fluid copper and Cp concentration sharply increased. The role of milk Cp in controlling the copper balance in newborn rats and the tissue-specific mechanism of Cp gene activity regulation are under discussion.

Anemia and impaired stress-induced erythropoiesis in aceruloplasminemic mice.

Ceruloplasmin (Cp) is an abundant, copper-containing plasma protein with an important role in iron homeostasis. Patients with hereditary Cp deficiency have iron deposits in liver and other organs, consistent with impaired iron flux. The mild anemia reported in some patients suggests a possible role for Cp in iron delivery to red cell precursors during erythropoiesis. To investigate this function of Cp, we determined the hematologic parameters in Cp-deficient mice under normal conditions and after erythropoiesis-inducing stress. Cp(-/-) mice have below normal hematocrit, red cell hemoglobin and volume, and serum iron. Red cell number and turnover and reticulocyte counts were identical in Cp(-/-) and Cp(+/+) mice. Thus, Cp(-/-) have mild microcytic, hypochromic anemia consistent with normal red cell formation but defective iron availability. Cp(-/-) and Cp(+/+) mice subjected to phenylhydrazine-induced hemolytic anemia exhibited identical decreases in hematologic parameters, but Cp(-/-) mice showed diminished recovery after removal of the stress. Administration of purified human Cp or iron-saturated transferrin to Cp(-/-) mice partially restored hemoglobin formation in reticulocytes. The mild anemia in Cp(-/-) mice and the diminished response to stress may reflect inefficient recycling of iron between the reticuloendothelial and erythropoietic systems. Our findings suggest a role for Cp in erythropoiesis by providing sufficient iron to the erythroid tissue and that the requirement for Cp is raised after erythropoietic stress.

[Antioxidant and antihypoxic effects of the ceruloplasmin preparation in the hypobaric hypoxia model]. / Izuchenie antioksidantnogo i antigipoksicheskogo deistviia preparata tseruloplazmin na modeli gipobaricheskoi gipoksii.

The antioxidant and antihypoxant effects of the drug ceruloplasmin were studied on an experimental model of hypobaric hypoxia in rats. Two forms of the drug characterized by low and high enzymatic activity were administered (intramuscularly or intraperitoneally) 20 min or 1 day before hypobaric hypoxia test. The drug effect was independent of the enzymatic activity level. The maximum activity of ceruloplasmin was observed when the drug was administered one day before test. Analyses showed that the drug activated superoxide dismutase, while not changing the catalase activity.

[Characteristics of expression of active oxygen forms by blood cells in patients with chronic bronchitis]. / Osobennosti ékspressii aktivnykh form kisloroda kletkami krovi u bol'nykh khronicheskim bronkhitom.

AIM: To study generation of active oxygen forms in the whole blood of patients with chronic bronchitis and correction of the detected disorders. MATERIAL AND METHODS: By means of registration of spontaneous and staphylococcus-induced luminol-dependent chemiluminescence (LDCL) of the whole blood, generation of active oxygen forms in 73 patients with chronic bronchitis (CB) has been studied. RESULTS: By the intensity of luminescence, all the examinees were divided into two groups. In group 1 LDCL was high, in group 2 LDCL was under the control level. Combined treatment of group 1 patients included inhalations of ceruloplasmin antioxidant. Patients of group 2 were administered sodium hypochlorite that possesses prooxidation properties. Compared to conventional treatment, the above two treatments produced marked positive changes in the studied indices and clinical symptoms. CONCLUSION: The method of chemiluminescence registration of the whole blood allows validation of pathogenetic therapy of patients with CB and improves outcomes of the treatment.

Cu nutrition in infants during prolonged exclusive breast-feeding: low intake but rising serum concentrations of Cu and ceruloplasmin.

We studied Cu intake and nutrition in exclusively breast-fed infants by determining Cu and ceruloplasmin concentrations in maternal and infant sera, as well as milk intakes and concentrations. The infants numbered 200 at birth, 116 at age 6 mo, 36 at 9 mo, and 7 at 12 mo. Postpartum the mean maternal serum concentrations of Cu and ceruloplasmin were high, but decreased in 4 mo to the level of nonpregnant women, and remained thereafter stable. The median milk Cu concentration decreased throughout lactation. In contrast, the mean infant serum concentrations of Cu and ceruloplasmin increased with age reaching adult levels by age 6 mo. The infant serum concentrations were independent of the milk concentrations which in turn were independent of the maternal serum concentrations and the degree of maternal supplementation (none, 2, or 4 mg Cu++ with Fe++ and Zn++). Neither maternal nor infant serum concentrations reflected intake of Cu. The daily Cu intake varied up to 10-fold between infants. No signs of Cu deficiency were detected during exclusive breast-feeding.

Clinical results on the use of human ceruloplasmin in aplastic anemia.

Frequent blood transfusions are necessary in aplastic anemia because of decreased hematopoietic function. In order to maintain the hemoglobin level and to decrease the need for blood transfusion, human ceruloplasmin was used for the treatment of 73 patients with aplastic anemia. A marked beneficial effect was obtained in 16 cases. The treatment was moderately effective in 17 cases, slightly effective in eight cases, and ineffective in 32 cases. The usual dose of ceruloplasmin was 15 mg/day, but it was varied according to symptoms. Side effects were minimal when ceruloplasmin was administered by slow intravenous injection.

Effects of copper deficiency and copper loading on 67Cu in supernatants of rat organs.

Eleven organs of the rat were studied 1 hr and 24 hr after the intravenous administration of 67CuCl2 and 67Cu-ceruloplasmin. The rats were normal, copper-deficient, or copper-laden. The amounts of stable copper and 67Cu in the whole organ and supernatant fractions, corrected for whole blood copper and 67Cu, were measured. The distribution of supernatant 67Cu was determined in three Sephadex G-100 chromatographic zones: Peak I (150,000 daltons), Peak II (31,000 daltons), and Peak III (11,000 daltons). All organs took up 67Cu from both sources, but there was a tendency for increased uptakes in copper-deficient rats and decreased uptake in copper-laden rats. Only lung, heart, and testis took up more 67Cu from 67Cu-ceruloplasmin than from 67CuCl2. Supernatant 67Cu tended to be in Peak I when the source was 67Cu-ceruloplasmin and in Peak II when the source was 67CuCl2. When 67Cu-ceroloplasmin was added to supernatant fractions in vitro, the 67Cu was in Peak I. However, when 67CuCl2 was added to supernatant fractions, Peak III predominated in kidney, brain, testis, and liver; Peak II predominated in none; and Peak I predominated in spleen, muscle, large and small bowel, stomach, lung, and heart. A high-molecular-weight copper-binding substance seems to be present in organ supernatant fractions.