Identification of a myeloperoxidase inhibitor from normal human serum.

An inhibitor of myeloperoxidase (MPO) has been identified in normal human serum. Initial experiments confirmed that high levels of MPO inhibitory activity are present in sera and that the inhibitor did not act by interfering with the assay. Purification of the inhibitor activity by salt precipitation followed by ion exchange and affinity chromatography revealed the presence of a protein of 150 kD. The purified inhibitory activity displayed dose and time dependency and was not associated with IgG or IgA. It is considered that human serum contains an inhibitor of extracellular MPO capable of protecting against hypohalous acid release in host tissues and that if inhibitor levels are reduced such protection may fail.     

Assessment of the protective action of diphosphonate compounds against damage to T-lymphocytes by antilymphocytic serum

Rabbit antilymphocytic serum (ALS) and complement were taken in doses causing death of 50% of lymphocytes (stained with 1% trypan blue solution). The number of rosette-forming and transformed cells in the rosette-formation and blast-transformation tests was 60 and 24% of the number in the control respectively. After addition of 0.1 ml of 10 mM solution of the potassium salt of hydroxyethylenediphosphonic acid, belonging to the group of synthetic diphosphonates, a protective effect was observed: The number of rosette-forming and transformed lymphocytes did not differ from the control, in which the test was carried out with intact cells.Moscow Research Institute of Pediatrics and Pediatric Surgery, Ministry of Health of the RSFSR. (Presented by Academician of the Academy of Medical Sciences of the USSR P. N. Kosyakov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 3, pp. 259–260, March, 1979.     

Surface proteins of schizont-infected erythrocytes and merozoites of Plasmodium falciparum

Schizont-infected erythrocytes and merozoites were isolated from in vitro cultures of the human parasite, Plasmodium falciparum labeled with various radioactive substrates. The isolated merozoites were viable since they were able to reinvade fresh erythrocytes. On the basis of sensitivity to specific enzymes, eleven proteins synthesised by the parasite, were localised on the surface of the schizont-infected erythrocyte. Eight of these were glycoproteins, six of which appeared to represent three doublets. Five merozoite surface proteins were identified on the basis of their sensitivity to trypsin and chymotrypsin, treatments which also rendered the merozoite incapable of erythrocyte invasion. Merozoites appeared not to contain any glycoproteins; all of the glycoproteins synthesised by the parasite were apparently transported to the surface of the schizont-infected erythrocyte.     

Peculiarities of hemoglobin interaction with serum proteins of mice with Ehrlich carcinoma

In male C57B1/6 mice with transplanted Ehrlich carcinoma, hemoglobin forms a complex with serum proteins characterized by a molecular weight of about 300 kDa. The complex incorporates proteins weighing 100, 68, 65, and 15 kDa identified by MALDITOF mass spectrometry as haptoglobin, serum albumin, gi|26341396 nameless protein Mus musculus, and α-hemoglobin, respectively. This complex can possess biological activity and contribute to the control of tumor growth. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 141, No. 5, pp. 563–566, May, 2006     

Isoelectric focusing and immunochemical analysis of proteins of the “Diaferm-3” antitetanus serum

The isoelectric points (PIs) of proteins present in the Diaferm-3 antitetanus serum, a mixture of partially purified [F'ab]₂-fragments of native molecules of antitetanus immunoglobulins, their antitoxic activity, and their immunochemical properties were investigated by isoelectric focusing in a pH gradient of ampholines. The PI values and electrophoretic mobility of the preparation were compared. Proteins with antitoxic activity were shown to have PI values within the range 9.1–5.15, and from their electrophoretic mobility they can be classed as G- and -globulins. No antitoxic activity was found in macroglobulins with the mobility of -globulins.Laboratory of Pathophysiology of Toxicoinfections, Institute of Normal and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 10, pp. 41–45, October, 1974.     

Identification of specific serum proteins synthesized de novo by monolayer cultures of glandular cells of gestational endometrium

Monolayer cell cultures (n = 3) of glandular epithelium of gestationalendometrium obtained from three apparently healthy women undergoingelective termination of pregnancy (7–9 weeks gestation)were established. De novo synthesis of eight serum proteins(albumin, alpha₁-antitrypsin, cerulo-plasmin, beta-lipoprotein,alpha₂-macroglobulin, fibronectin and complement factors C3and C4) was demonstrated by the incorporation of radiolabelledsubstrate ([³⁵-S]methionine) employing autoradiography (AR)in combination with crossed immunoelectrophoresis (XIE), referredto as ARXIE, and line immunoelectrophoresis (LIE), referredto as ARLIE. By contrast, there was no evidence for de novosynthesis of IgA, haptoglobin and orosomucoid. Our findingssuggest that the gestational endometrium may contribute to theproduction of several proteins considered to be synthesizedand secreted mainly by the liver and reticulo-endothelial system.The simple techniques used here to identify the de novo synthesisof human serum proteins could be applied to investigate proteinsynthesis by a wide range of tissues and cells     

Protective action of some diphosphonates against injury to lymphocytes by antilymphocytic serum

Rabbit antilymphocytic serum and complement were used in quantities causing death of 50% of human lymphocytes isolated in a Ficoll-Verografin (amidotrizoate) density gradient. The experimental samples (0.2 ml of a lymphocyte suspension containing 2.4×10⁴–6×10⁴ cells) were treated with 10 mM solutions of diphosphonates [disodium salt of hydroxyethylenediphosphonic (HEDP) acid, alkylated HEDP-acid, aminomethyl-HEDP-acid, aminobenzyldiphosphonic and aminoisopropyldiphosphonic acids] in doses of 0.001 to 0.2 ml. A decrease in the number of dead cells was observed after staining with 0.1% trypan blue. The disodium salt of HEDP-acid and alkylated HEDP-acid proved to be most effective and exhibited protective properties in doses as low as 0.01 ml. In a dose of 0.1 ml, all the tested compounds had a marked protective action.Research Institute of Pediatrics and Pediatric Surgery, Ministry of Health of the RSFSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR, P. N. Kosyakov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 9, pp. 339–341, September, 1978.     

Oxidative myocardial damage: Protective action of exogenous phosphocreatine

Laboratory of Bioenergetics of the Heart, Institute of Experimental Cardiology, All-Union Cardiologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. N. Smirnov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 109, No. 3, pp. 224–226, March, 1990.     

Effect of seal serum proteins on the analgesic action of narcotic drugs

Research Laboratory of Biologically Active Substances of hydrobionts, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR P. V. Sergeev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 11, pp. 503–505, November, 1991.     

Covalent coupling of proteins to erythrocytes by isocyanide. A new,sensitive and mild technique for identification and estimation of antibodies by passive hemagglutination

A mild method for covalent coupling of protein-antigens to the surface of red blood cells (RBC) is described. The method is based on the formation of covalent bonds between aldehyde groups of oxidized RBC and amino or carboxyl groups of proteins in the presence of isocyanide.Very low concentrations of antibodies (8–30 ng/ml) could be detected in antisera with the specifically sensitized RBC as reagents in passive hemagglutination with no or very low non-specific agglutination.     

Separation of Fc-binding serum proteins by preparative flat-bed isotachophoresis

Preparative flat-bed isotachophoresis with discrete spacers was applied as a single-step procedure to separate 2 Fc-dependent activities of normal chicken serum, i.e., (1) the ability to raise the titre of haemagglutinating allo-antisera which is due to a high molecular weight beta-globulin (HEF), (2) the ability to activate guinea pig complement components in mixed complement reaction. The results demonstrate that the 2 activities can be clearly separated, and HEF must therefore be different from the first complement factor in the chicken. Under the chosen conditions the molecule active in the mixed complement reaction is not stacked in contrast to other serum protein including HEF. The same technique with human serum shows that human Clq behaves in the same was as the chicken complement factor. This means that by selective unstacking, flat-bed isotachophoresis can be used as an efficient single-step purification method for human and chicken Clq.     

Effect of hyperoxia and the protective action of urea on serum hemoglobin,transferrin, and total iron concentrations

During exposure of albino rats to oxygen under a pressure of 4 atm for 1 h (the preconvulsive state) and of 6 atm (convulsive state) an increase in the serum hemoglobin concentration was found. The total ion concentration increased at the same time. By disc electrophoresis in 7.5% polyacrylamide gel, changes in the ratio between the hemoglobin fractions in the serum were found in both stages of oxygen poisoning. An increase in the concentration of transferrins was found during hyperoxia under these conditions. If urea was administered to the animals before the session of hyperbaric oxygenation, the changes observed were less marked.Department of Biochemistry, Rostov-on-Don University. (Presented by Academician of the Academy of Medical Sciences of the USSR V. N. Chernigovskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 6, pp. 528–531, June, 1979.     

Conglutination and haemolysis of unsensitized human erythrocytes by bovine serum complement

Unsensitized human erythrocytes (E) were haemolyzed by bovine serum to a titre of 1:16-1:32. In the single dilution beyond the haemolytic endpoint, the cells were conglutinated. In dilutions in which haemolysis occurred, cells were conglutinated before being lyzed. With no or minimal haemolysis, conglutination to 1:32-1:64 occurred in tests using insulin-absorbed serum, serum heated at 50 degrees C for 30 min and in tests incubated at 4 degrees C for 30 min. In two-stage tests, EDTA and Mg2+-EGTA prevented bovine C sensitization of human E for conglutination by bovine serum heated at 56 degrees C for 30 min. EDTA prevented haemolysis, but haemolysis to 1:16-1:32 occurred with serum dilutions containing Mg2+-EGTA. Haemolytic activity was restored to serum heated at 50 degrees C by a factor B-containing fraction. Conglutination and haemolysis were blocked by heating serum at 56 degrees C for 30 min and were reduced to low titres by absorbing serum with zymosan. These results strongly suggest that the conglutination reaction involved the classical activation pathway whereas the haemolytic reaction involved the alternative activation pathway. Thus, with dilutions of untreated or treated bovine serum, two C-dependent reactions and the pathways involved can be demonstrated by using unsensitized human E as an indicator system.     

Sex differences in blood protein patterns: a computer-assisted electrophoretic analysis of mid-molecular weight range proteins of human serum

We have studied the sexual dimorphism of human serum proteins between the ranges of approximately 60,000 and 150,000 daltons using computerized imaging and densitometry. Methodology is presented, based on the TEXAC computer system for the detection, segmentation and integration of protein bands. The results are discussed in terms of potential future applications in computerized medical diagnosis.     

Oxidative damage of lung and its protective mechanism in mice caused by long-term exposure to titanium dioxide nanoparticles

Exposure to titanium dioxide nanoparticles (TiO(2) NPs) elicits an adverse response such as oxidative damage. The molecular targets of TiO(2) NPs remain largely unidentified. In the present study, the function and signal pathway of nuclear factor erythroid 2 related factor 2 (Nrf2) in protection against TiO(2) NPs-induced oxidative stress in the mouse lung were investigated. Mice were exposed to 10 mg/kg body weight by an intratracheal administration for 15-90 days. With increasing exposed terms, TiO(2) NPs were significantly accumulated and increased the reactive oxygen species (ROS) production in lung, which resulted in severe pulmonary edema, inflammatory response and pneumonocyte apoptosis for 90 days. Furthermore, TiO(2) NPs exposure could greatly induce expression of Nrf2, heme oxygenase 1 (HO-1), and glutamate-cysteine ligase catalytic subunit (GCLC) from 15-day to 75-day exposure, whereas 90-day exposure caused significant decreases of three factors expression levels in lung. Our findings imply that the induction of Nrf2 expression is an adaptive intracellular response to TiO(2) NPs-induced oxidative stress in the mouse lung, and that Nrf2 is protective against TiO(2) NPs-induced pulmonary damages during certain exposure terms. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A 100A:2554-2562, 2012.