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Biomedical subjects

J Rejnek

Publications and source records attributed to J Rejnek.

At least 19 recordsLinked to original sources

Iron humate as a low-cost sorbent for metal ions.

Iron humate produced as a waste by-product during an industrial manufacture of humic substances from low-rank brown coals was tested as a sorbent for the removal of metal cations--Cd (II), Cu (II), Co (II), Ni (II), Zn (II), TI (I), Eu (III), Cr (III)--as well as hexavalent chromium from waters. The Langmuir-type isotherms were used to describe the metal sorption; the respective equations may be derived and interpreted on the basis of a concept of surface-complexation reactions. Parameters of the sorption isotherms were estimated from experimental dependencies measured in a batch arrangement. The sorption capacities ranged from 0.024 mmol g(-1)for Co (II) to 0.324 mmol g(-1)for Tl(I). The metal uptake was affected by the presence of complexing agents (EDTA, salicylate, citrate), but this effect depends strongly on pH and the kind of complexing agent. The sorption of Cu (II) ions was suppressed in the presence of EDTA in an almost whole examined pH range (ca. 1-6.5) and in the presence of citrate at higher pH values (above pH 4). In the presence of salicylate, on the other hand, a slight sorption enhancement was observed. The metal sorption was suppressed also in the presence of anionic surfactant (sodium dodecylsulfate). Both in the presence as well as in the absence of complexing agents, the metal sorption showed a strong dependence on pH--the sorption was reduced considerably at low pH values below ca. 2. A typical working range for iron humate as a sorbent is ca. pH 3-5, where it exhibits a high buffering capacity, a sufficient stability and metal-binding capability.

Adsorption↗

The fate of protein antigen in earthworms: study in vivo.

Parenteral administration of antigen induces in the earthworms Lumbricus terrestris and Eisenia foetida the formation of antigen-binding protein. Labelled antigen was efficiently digested in the coelomic fluids of both species since the total amount of radioactivity in the coelomic fluids decreased. Antigen internalized in the free coelomocytes of both species was present in a TCA-precipitable form, although the decline of total radioactivity was higher than in the coelomic fluid, indicating that the TCA-nonprecipitable fragments are rapidly released from coelomocytes into the coelomic fluid. Within the first 24 h after administration, labelled antigen was detected throughout all tissues except for the epidermis and cuticle, and after 4 days it was found in the chloragogenic tissue around the gut, dorsal vein and typhlosole.

Animals↗

The fate of protein antigen in earthworms: study in vitro.

Earthworm coelomocytes digest protein antigen in vitro. Proteolytic activity was detected both in cell-free coelomic fluid and in cell cultures of free coelomocytes which are effectors of earthworm immunodefense mechanisms. Antigen is cleaved either intracellularly or by proteolytic enzymes released by coelomocytes into the medium. Proteolysis was observed both in non-stimulated and antigen-stimulated cultures. Since significantly higher proteolysis was shown in supernatants from cultures of antigen-stimulated coelomocytes, we can assume that the release of proteolytic enzymes was inducible.

Animals↗

Role of saccharides in immunoglobulin--Fc receptor interactions.

The rosettes formed by mouse peritoneal macrophages or DCH-5 cells and TNP-erythrocytes coated with anti-TNP antibodies of different isotypes were inhibited to various extent by monosaccharides. The most effective inhibitors were N-acetylglucosamine, glucosamine, mannose and N-acetylneuraminic acid in 1-5 mmol/L concentrations. Even more efficient were glycopeptides isolated from IgG molecules. The Fc receptors (FcRs) released from DCH-5 cells during cultivation and gradually separated by affinity chromatography on immobilized IgG reacted with aggregated IgG and inhibited the rosette formation. The FcRs eluted by monosaccharides influenced mainly the number of rosettes mediated by IgA and IgE while those eluted with a glycine-HCl buffer inhibited preferentially IgG rosettes. As shown by SDS-PAGE the heterogeneity of the fraction eluted with a mixture of monosaccharides revealed one main component with an effective molar mass of 50 kg/mol. The glycine-HCl eluate contained two major components of 55 and 38 kg/mol. The IgG-Sepharose 4B bound all the fractions but only the binding of the 50 kg/mol molecule could be inhibited by monosaccharides.

Animals↗

Characterization of antigen-binding protein in earthworms Lumbricus terrestris and Eisenia foetida.

Injection of antigen into the annelid worms Lumbricus terrestris (LT) and Eisenia foetida (EF) results in a marked increase of coelomic fluid protein concentration and the formation of a protein which binds the stimulating antigen (3). In this report we show that the increases in total protein concentration after first and second doses of antigen were higher and were achieved earlier in LT than in EF, while the accumulation of antigen-binding protein in coelomic fluid was similar in both species. Antigen-binding protein isolated by affinity chromatography retained its original binding activity. Its molecular weight in coelomic fluid as well as after isolation was 56 kD when analyzed by SDS-PAGE and immunoblotting. Under reducing conditions, two bands with mol./wt. 31 and 33 kD appeared which did not reveal detectable binding activity. This suggests that the 56 kD binding protein of annelids is composed of two disulphide-linked polypeptide chains both of which participate in antigen-binding site formation.

Animals↗

The interaction of a protein from the coelomic fluid of earthworms with staphylococcal protein A.

A Staphylococcal protein A (SpA)-binding protein was isolated from coelomic fluids of the annelids, Lumbricus terrestris (LT) and Eisenia foetida (EF), by affinity chromatography on SpA-Sepharose. Analyses by polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE) under reducing and nonreducing conditions showed that SpA-binding activity is associated with a single chain protein, with a mol wt of 62 kD. The carbohydrate moiety of this protein consists of 21.4 g/100 g of O-linked and N-linked oligosaccharides. The amino acid composition of SpA-binding protein did not show structural homology with that of human IgG1 heavy chain (Daw), which also binds SpA.

Amino Acids↗

Detection of antigen in the coelomocytes of the earthworm, Eisenia foetida (Annelida).

Earthworms, Eisenia foetida, are able to respond to antigenic stimulation by the formation of the antigen-binding molecules by coelomocytes--the effector cells of annelids' defence reactions. The ability to react with gold-labelled antigen was detected in agranular coelomocytes by electron microscopy. Furthermore, flow cytometry analysis used for quantitative evaluation of antigen binding showed significant increase of both antigen-binding cells and the amount of antigen bound per cell after stimulation. The antigen binding was inhibited by preincubation of cells with several similar proteins, although the most potent inhibitor was the immunizing antigen.

Animals↗

In vitro antigen-binding properties of coelomocytes of Eisenia foetida (Annelida).

Annelids are capable of cellular and humoral defence reactions against foreign antigens. The main aim of this study was to characterise the antigen-binding properties of coelomocytes of Eisenia foetida by means of quantitative autoradiography and direct measurement of radioactivity. It was found that the antigen-binding capacity was significantly increased after antigen stimulation. Furthermore, the preincubation of coelomocytes with non-labelled proteins reduced the binding of radiolabelled antigen. The highest level of inhibition was found when the same protein was used for preincubation. These results indicate that antigen-binding properties are to some extent specific.

Animals↗

Response to parenteral stimulation in earthworms L. terrestris and E. foetida.

Injection of different protein molecules into the coelomic cavity of Eisenia foetida (E.F.) and Lumbricus terrestris (L.T.) markedly stimulated proteosynthesis. A similar but less pronounced increase of coelomic proteins was induced by phosphate buffered saline (PBS). In the coelomic fluids of stimulated animals a protein was detected which significantly bound to the 125I or peroxidase labelled protein molecules used for stimulation. The maximal binding occurred on day 8 after primary stimulation. A greater increase was observed after secondary stimulation. The degree of binding specificity appeared to be much lower than that of vertebrate lymphocyte receptors or antibodies.

Animals↗

Lytic activities in coelomic fluid of Eisenia foetida and Lumbricus terrestris.

Coelomic fluids of the two earthworm species E.foetida (E.F.) and L.terrestris (L.T.) have not only the ability to lyse various vertebrate erythrocytes but also to digest vertebrate serum proteins. Both activities are carried by different molecules since hemolysis but not proteolysis was inhibited by simple sugars. In contrary, proteolysis was blocked by PMSF which did not influence hemolysis. Coelomic fluids of E.F. digest effectively vertebrate serum proteins (PIgG, HSA) but not the proteins of L.T. coelomic fluids. The proteolytic activity was detected in approximately 40 000 mol. wt. fraction. After digestion proteolytic fragments were analyzed by immunoelectrophoresis, SDS-PAGE and TCA precipitation. Two of the fragments reacting with PIgG antisera remained intact even after 120 h digestion.

Animals↗

The proteins in Lumbricus terrestris and Eisenia foetida coelomic fluids and on coelomocytes reacting with sheep and goat IgG molecules.

Sheep and goat fast migrating IgG molecules and their F(ab')2 fragments react with L. terrestris and E. foetida coelomic fluid proteins. The reaction was not mediated by antibody binding site but by some other part of the IgG molecule, since affinity purified antibodies of different antigen specificities as well as IgG preparations from normal sheep sera reacted with the same coelomic fluid proteins. The presence of at least two reacting proteins was detected by immunoelectrophoresis and immunoblotting analysis. Molecular weights of these proteins were 47 and 50 kD in L. terrestris and 42 kD in E. foetida. Sheep antibodies and IgG molecules also bind to the Sheep antibodies and IgG molecules also bind to the surface of about 2% of coelomic leukocytes and the immunoblotting analysis showed that the IgG binding proteins in coelomic fluids and in cell lysates were similar.

Animals↗

Biochemical characterization of Fc gamma receptors of mouse DCH-5 cell line.

Fc gamma receptors (Fc gamma R) were isolated from the culture medium of a mouse hybrid cell line (DCH-5) known as an over-expressor of Fc gamma R, established from a mouse adherent spleen cell and the thymoma cell BW 5147. Proteins adsorbed to insolubilized IgG were separated on Sephacryl S-200. The main fraction with maximum FcR activity was isolated and characterized as a glycoprotein with an effective molar mass of about 55 kg/mol. Under non-denaturing conditions, the protein existed as a non-covalently linked dimer. Isoelectric focussing in agarose gel showed two bands with pI = 5.2 and pI = 5.3. The amino acid composition of this fraction was similar to that of pig and human Fc gamma R, and also of rabbit FcR for polymeric Ig. The sugar composition of the fraction (about 34% w/w) resembled that of the C1q component of complement and some membrane glycoproteins.

Amino Acid Sequence↗

Antigenic properties of T cell antigen-specific receptors isolated from the surface of rabbit and mouse spleen and lymph node cells.

The presence of a allotypic determinants was tested in fractions obtained by gel filtration of antigen-specific receptors isolated by immunoadsorption from lymphoid cells of antigen-stimulated a3-3 rabbits. This technique, as well as the inhibition of the reaction of isolated receptors with anti-T cell receptor antisera (anti R) by anti-a3 antibodies failed to demonstrate the presence of a allotypic determinants. The inhibitory effect of antigen-specific receptors isolated from the lymphoid cells of stimulated A/J mice on the cytotoxic effect of anti-Ia antibodies on mouse spleen cells in the presence of rabbit complement was tested. All preparations inhibited the cytotoxic reaction with the average effectivity of 60%. In order to confirm the presence of Ia determinants on the rabbit and mouse T cell receptor molecules it was shown that the reactions of three anti-R antisera with 12 different receptor preparations were inhibited by anti-Ia antibodies. SDS-PAGE analyses of 125I-labelled mouse specific receptors and the precipitate obtained by anti-R antisera showed that T cell receptors were present in fractions with molar mass 100 and 85 kg/mol. The molar mass of the former fraction after reduction and alkylation was 45 kg/mol.

Animals↗

Influence of proteolysis inhibitors and immunoadsorption technique on the composition of rabbit antigen-specific receptor preparations. Presence of Fc receptors.

Addition of proteolysis inhibitors during the isolation procedure of rabbit arsanylated bovine IgG specific receptors decreased significantly the amount of low-molar-mass proteins in all receptor preparations. Rabbit ARS-BGG-specific receptor preparations isolated by immunoadsorption technique contain molecules which do not react with antigen and antibodies against immunoglobulins and have identical molar mass and chymotryptic peptide composition as those of isolated Fc receptors. It is suggested that during isolation of antigen-specific receptors from the surface of lymphoid cells, Fc receptors react with complexes composed of antigen and Ig+ receptors on the surface of immunoadsorbent and are isolated together with antigen-specific receptors.

Animals↗