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H Kaltner

Publications and source records attributed to H Kaltner.

71 records · Page 4Linked to original sources
PubMed1996

Binding capacities of two immunomodulatory lectins, carrier-immobilized glycoligands and steroid hormones in lung cancer and the concentration of nitrite/nitrate in pleural effusions.

Combined analysis of the binding properties of inflammatory and tumor cells in pleural effusion, and tumor imprints for various carrier-immobilized types of ligands and lectins, and of a biochemical feature of the effusions is performed to extend the characterization of these cells and their activity. In detail, the binding of Viscum album agglutinin (VAA), Urtica dioica agglutinin (UDA), and of carrier-immobilized N-acetyl-D-glucosamine (GlcNAc), lysoganglioside GM1, estradiol, progesterone, testosterone, and hydrocortisone to native specimens consisting of 46 tumor imprints from surgically treated patients with lung cancer and 74 smears of pleural effusion (PE) cells from cancer or non-cancer patients was studied using fluorescence microscopy with Texas red-labeled streptavidin. Among the tested ligands, VAA was found to provide the most effective staining of cells (60-78.1% of positive cases). When compared with inflammatory cells from PE, cancer cells were seen to bind more frequently only two ligands, namely UDA and estradiol. Significant (P < 0.001) difference between patients with bronchial carcinoma and non-cancer patients were found, when the content of NO2-/NO3- in PE fluids was measured. Whereas the level of NO2-/NO3- in PE of non-cancer patients was 12.6 +/- 10.7 microM (n = 12), it was 37.7 +/- 19.4 microM (n = 14) in cancer patients without pleural metastases and 37.5 +/- 16.0 microM (n = 26) in patients with pleural metastases. The level of NO2-/NO3- in PE appeared to correlate with extent of staining with GM1 and GlcNAc: in non-cancer patient groups it was significantly higher (P = 0.032) for negative subjects than those binding the ligand GlcNAc, whereas in the patient group with adenocarcinoma it was significantly lower (P = 0.032) for patients without binding capacities for GlcNAc and GM1. The results obtained suggest that the combined analysis of increased levels of NO2-/NO3- in PE and of glycohistochemical properties of cancer and inflammatory cells may be useful in exploring the interrelationship of functionally important cellular characteristics.

Acetylglucosamine↗
PubMed1996

Comparative cross-linking activities of lactose-specific plant and animal lectins and a natural lactose-binding immunoglobulin G fraction from human serum with asialofetuin.

Plant and animal lectins bind and cross-link certain multiantennary oligosaccharides, glycopeptides, and glycoproteins. This can lead to the formation of homogeneous cross-linked complexes, which may differ in their stoichiometry depending on the nature of the sugar receptor involved. As a precisely defined ligand, we have employed bovine asialofetuin (ASF), a glycoprotein that possesses three asparagine-linked triantennary complex carbohydrate chains with terminal LacNAc residues. In the present study, we have compared the carbohydrate cross-linking properties of two Lac-specific plant lectins, an animal lectin and a naturally occurring Lac-binding polyclonal immunoglobulin G subfraction from human serum with the ligand. Quantitative precipitation studies of the Lac-specific plant lectins, Viscum album agglutinin and Ricinus communis agglutinin, and the Lac-specific 16 kDa dimeric galectin from chicken liver demonstrate that these lectins form specific, stoichiometric cross-linked complexes with ASF. At low concentrations of ASF, 1:9 ASF/lectin (monomer) complexes formed with both plant lectins and the chicken lectin. With increasing concentrations of ASF, 1:3 ASF/lectin (monomer) complexes formed with the lectins irrespective of their source or size. The naturally occurring polyclonal antibodies, however, revealed a different cross-linking behavior. They show the formation of 1:3 ASF/antibody (per Fab moiety) cross-linked complexes at all concentrations of ASF. These studies demonstrate that Lac-specific plant and animal lectins as well as the Lac-binding immunoglobulin subfraction from specific stoichiometric cross-linked complexes with ASF. These results are discussed in terms of the structure-function properties of multivalent lectins and antibodies.

Animals↗
PubMed1996

[The mistletoe myth--claims, reality and provable perspectives].

Intuition guided R. Steiner to refer to mistletoe as the future remedy for cancer. He proposed that its spiritual qualities support re-establishment of the harmonious integration of the alleged four different entities of human existence in a patient. The assumption of potency without chemical basis is derived from the dogmatic system of anthroposophic reasoning. It explains the evidently similar claim of clinical efficiency for the proprietary mistletoe extracts despite the lack of information on the actual contents of the complex mixtures and despite the conspicuous diversity of methods of manufacture for these products. Thorough scientific analysis of the published clinical experience does not justify this claim. Due to the increasing reference to defined substances in advertisements for commercial extracts, they should no longer avoid rigorous testing according to common quality standards. Interdisciplinary research efforts on the immunomodulatory galactoside-binding lectin illustrate how to yield a clinically testable substance from an ill-defined extract, thereby providing a notable example for rational investigation of unconventional treatment modalities.

Adjuvants, Immunologic↗
PubMed1995

Differential binding of two chicken beta-galactoside-specific lectins to homologous lymphocyte subpopulations and evidence for inhibitor activity of the dimeric lectin on stimulated T cells.

Plant lectins can be potent modulators of vertebrate immune functions. Biochemical characterization of lectins from animal tissues enables the determination of whether these endogenous activities display a comparable immunological potency. Focusing on chicken beta-galactoside-binding lectins, the monomeric intestinal (CL-14) and the dimeric liver lectin (CL-16) were purified and the lack of cross-contamination was ascertained. In very close agreement with the molecular masses of 14,974 and 14,976 calculated on the basis of the available sequence data (Y. Sakakura et al., J. Biol. Chem. 265, 21573-21579, 1990), electrospray mass spectrometric analysis yielded values of 14,969 (CL-14) and 14,972 (CL-16), the reasons for the deviation in gel electrophoretic behavior being unclear. Solid-phase assays with immobilized lactosylated poly-L-lysine demonstrated a comparatively lower affinity and higher extent of binding at saturation for the monomeric lectin than for the dimeric protein, whose properties were similar to those of an immunomodulatory plant lectin. Flow cytometry revealed homogeneous and strong binding of the dimeric lectin within the chicken peripheral blood lymphocyte population, whereas the monomeric lectin stained two subpopulations at different intensities. Two-color flow cytometry disclosed preferential binding of this lectin to B cells. When a B cell line was employed for determination of affinity constants and extents of binding at saturation, qualitatively comparable parameters to those for the solid-phase assays were obtained. The similar profile of lectin-binding glycoproteins in blots of cellular extracts underscored that accessibility to ligands, not qualitatively different ligand display, may explain the differences for the cell line. At up to a concentration of 10 micrograms/ml of the lectins no stimulation of [3H]thymidine incorporation was seen for blood and spleen cell populations. However, the dimeric lectin reduced stimulation of cells that were responsive to an anti-TcR2 antibody. Thus, this lectin can apparently exhibit inhibitory activity to this kind of T cell activation in vitro.

Animals↗
PubMed1995

Carbohydrate-binding proteins (plant/human lectins and autoantibodies from human serum) as mediators of release of lysozyme, elastase, and myeloperoxidase from human neutrophils.

Analysis of cell surface glycosylation not only provides information about cell properties such as their state of differentiation or histogenetic lineage. The carbohydrate chains also provide potentially functional binding sites to endogenous carbohydrate-binding proteins. This interaction can elicit consequent signalling processes. Because of the importance of neutrophils in the host defence system, we monitored the effect of the binding of such sugar receptors to their cell surface on the release of the enzymatic activities of lysozyme, elastase, and myeloperoxidase. Besides the mannose-binding lectin concanavalin A and the immunomodulatory alpha/beta-galactoside-binding lectin from Viscum album L., three preparations of human sugar receptors - beta-galactoside-binding lectin (M(r) 14 kDa) and two affinity-purified polyclonal IgG fractions from serum with the capacity to recognize alpha- or beta-galactosides, respectively - were used. Two animal lectins from chicken liver and intestine that bind beta-galactosides, as well as the lectin-like human serum amyloid P component, were included in order to assess the importance of slight differences in ligand recognition. Cytochalasin B-enhanced enzyme release was invariably seen with the two plant lectins and the chicken liver beta-galactoside-binding lectin, but the related intestinal lectin did not increase enzyme release. The mammalian homologue of these avian lectins triggered lysozyme secretion, and the lactoside-binding IgG fraction enhanced the amount of extracellular elastase activity slightly but significantly. Thus, the actual lectin, not the nominal specificity of sugar receptors, is crucial for elucidation of responses. Due to the highly stimulatory activity of the two plant lectins, neutrophils from patients with non-cancerous diseases and from patients with lung cancer were monitored for the extent of lectin-mediated enzyme release. Only the concanavalin A-mediated reactivity of the neutrophils was associated with the type of disease.

Adult↗
PubMed1995

Affinity-purified antibodies against alpha-galactosyl residues from human serum: comparison of their binding in bovine testicular tissue with that of the Griffonia simplicifolia lectin (GSI-B4) and impact of labeling on epitope localization.

alpha-Galactosyl residues in the carbohydrate part of cellular glycoconjugates can serve as cell type-associated markers and are implicated in intercellular adhesion and biosignaling. This biological significance explains the interest to characterize probes with respective specificity as the Griffonia simplicifolia I-isolectin B4. Due to the documented occurrence of an alpha-galactoside-binding immunoglobulin G fraction in human serum we compared the extent of binding and its pattern for the lectin and the antibody using surface-immobilized extract proteins and fixed sections of bovine testicular tissue with known lectin reactivity. The antibody fractions were obtained either solely from affinity chromatography isolation on immobilized melibiose or after an additional step to deplete this fraction of galactoside-binding activities without pronounced specificity to the alpha-anomeric linkage. They yielded a rather indistinguishable reactivity in comparison to that of the lectin, when an indirect approach was used. Labeling of the antibodies with a hydrazide derivative of biotin did not affect the pattern of binding. However, significant differences were noted, when conjugation of label was targeted to amino groups via N-hydroxy-succinimide esters of biotin and digoxigenin despite performance of the modification under activity-preserving conditions. Notably, the apparent strong staining of Leydig cells and nuclei of primary spermatocytes, respectively, was not inhibitable by sugar. These differences were corroborated by a nonidentical response of the various probes in solid-phase assays with extract proteins. Thus, care should be exercised in the interpretation of histochemical data, obtained with this type of modified antibody. When these precautions are fulfilled, this immunoglobulin fraction from human serum has the potential as an alpha-galactosyl-specific histochemical tool.

Animals↗
PubMed1994

[Lectin-based therapy applications from the laboratory to practice].

Thorough analysis of the principles of molecular recognition is the basis for rational development of clinical applications. Currently, our knowledge is expanding, how biological information is encoded in a language of carbohydrate moieties, constituting the glycopart of cellular glycoconjugates. Carbohydrate-binding proteins like lectins can specifically bind these ligands. This glycobiological interplay participates in recognitive inter- and intracellular processes that enable to devise clinical schemes with rational perspective like targeted drug delivery, non-steroidal treatment of inflammation or lectin ligand-dependent treatment of infectious diseases. Besides the ligands, lectins, too, can be of therapeutical value, e.g. as biomodulators in the immune system. The rapid development within glycobiology allows to propose that certain aspects can well find their place in veterinary practice after proving their efficacy in clinical trials.

Animals↗
PubMed1993

Activity of cholinesterases in the Japanese quail embryo. Effects of dichlorphos on the embryonic development.

Cholinesterase activity is detectable in the Japanese quail embryo, in the yolk and subembryonic liquid, but not in the albumen. Obviously, this enzyme is deposited by the hen into the yolk and from there it is transferred to the subembryonic liquid. In contrast, in the embryo the enzyme is synthesized by itself and the amount increases with the age of the embryo. By using BW284c51 1,5-bis-(4-allyldimethylammoniumphenyl)pentan-3-one bromide and ISO-OMPA tetraisoprophylpyrophosphoramide as inhibitors, it was found that the enzyme in the embryo is predominantly acetylcholinesterase (EC 3.1.1.7), whereas that in the yolk and subembryonic liquid is butyrylcholinesterase (EC 3.1.1.8). Both types are inhibited by dichlorphos. However, the embryonic enzyme activity is restored within 8 hr, whereas that in the subembryonic liquid remained inactive at least for 72 hr after inhibition. Enzyme inhibition leads to retardation of the development, to reduced accumulation of glucose and amino acids in the subembryonic liquid and finally to death of the embryo, suggesting that the developmental retardation is due to the restricted supply of glucose and amino acids. Surprisingly, most of the embryos die when the embryonic enzyme activity has again been restored.

Amino Acids↗
PubMed1993

Developmental retardation of the Japanese quail embryo under the influence of dexamethasone.

Cultured Japanese quail embryos were treated with various doses of dexamethasone on day 2 and in part also on day 5. The treatment lead to a significant delay of development. This was demonstrated by means of developmental (growth rate, developmental stage), histological and metabolic (excreted uric acid, protein synthesis) parameters. The effect was intensified when the agent was administered into the subembryonic liquid instead of the albumen. This is due to the fact that a higher dilution is expected when the agent is deposited into the albumen. In conclusion Japanese quail embryos respond to exogenous glucocorticoids, at a very early stage of development and at a comparatively low concentration with a depression of development.

Animals↗
PubMed1991

The influence of thiourea on the development of the cultured quail embryo.

Although thyroid hormones are known to promote embryonic development, it is still questionable whether the hormones of the thyroid gland exert a stimulatory effect on the early embryo. In attempting to elucidate the thyroid function in this period, we tried to block the synthesis of thyroid hormones by thiourea and followed the embryogenesis under these conditions. The experiments were performed with cultured quail embryos (Coturnix coturnix japonica), which seemed especially suitable for those studies. The results demonstrate that thiourea causes a decrease of the survival rate, DNA-synthesis and of the excretion of uric acid into the allantoic liquid. Furthermore, it exerts an inhibitory effect on weight gain and developmental stages. On the basis of these results it seems very likely, that thyroid hormones exert a stimulatory effect on the development of quail embryos earlier than assumed previously.

Animals↗
PubMed1988

Formation and changes of the subembryonic liquid from turned, unturned, and cultured Japanese quail eggs.

Japanese quail eggs belonging to the same flock of hens were incubated under different conditions: group 1 eggs were turned 3 times a day, group 2 eggs were left unturned, and group 3 eggs were cultured and left unturned. The results indicate that failure to turn eggs results in a delayed efflux of liquid and glucose from albumen and from the subembryonic liquid. Furthermore, the major difference between unturned and cultured eggs was that in the first group the glucose levels and in the second group the lactate levels of the subembryonic liquid were increased. It is suggested that reduced glucose supply may be involved in the disturbance of development of unturned and cultured eggs.

Animals↗
PubMedDate not supplied

Further refinement of the description of the ligand-binding characteristics for the galactoside-binding mistletoe lectin, a plant agglutinin with immunomodulatory potency.

The galactoside-binding lectin from mistletoe (Viscum album L.) is a biological response modifier, eliciting e.g. enhanced secretion of cytokines. This immunological activity warrants the further analysis of its ligand-binding properties with special attention paid to blood group epitopes. To avoid the microheterogeneity and complexity of naturally occurring glycoproteins, chemically strictly defined neoglycoconjugates and a panel of synthetic oligosaccharides were employed in solid-phase assays for direct binding and assessment of the relative inhibitory capacity. Since label incorporation into the lectin, although performed under protective conditions, or surface immobilization by adsorption to plastic may affect its affinity characteristics, the extent of neoglycoconjugate binding in the absence of any interfering substance and in the presence of oligosaccharides was determined comparatively with labeled and with immobilized lectin. In principle, these two factors could be excluded to markedly alter binding features. In addition to lactose, the blood group determinants H and B were strongly reactive. A fucose residue can thus especially be accommodated to the binding site when linked to the non-reducing unit. N-Acetyllactosamine was nearly as potent as an inhibitor as lactose. Lec and the A determinant were notably inferior to the other ABH blood group epitopes. Le(a) and Le(x) and their sialylated derivatives displayed only very weak binding capacity. Among the two natural isomers of sialyllactose, the alpha 2,6-form displayed a higher level of inhibitory capacity than the alpha 2,3-derivative. Isomeric variants of the Thomsen-Friedenreich antigen, too, reduced lectin binding to the lactose-carrying polymer. Their capacities were surpassed by those of the H and the B determinants and a related form of the latter, the P1 epitope. An overlap of specificity with the immunomodulatory human galectin-3 is thus measurable for H/B-like structures. The documented differential reactivity of the mistletoe lectin to blood group oligosaccharides may have a bearing on the responsiveness of blood group-positive cell populations.

Adjuvants, Immunologic↗
PubMedDate not supplied

Galectins-1 and -3 and their ligands in tumor biology. Non-uniform properties in cell-surface presentation and modulation of adhesion to matrix glycoproteins for various tumor cell lines, in biodistribution of free and liposome-bound galectins and in their expression by breast and colorectal carcinomas with/without metastatic propensity.

Protein (lectin)-carbohydrate (cellular glycoconjugate) recognition is operative in biochemical information transfer. Galectins constitute a family of endogenous galactoside-binding lectins with conserved features in the binding site. The members of this lectin category are assumed to be involved in cell adhesion and growth regulation. To assess to what extent the different modes of binding-site presentation and/or carbohydrate fine-specificities will affect aspects of galectin behavior, homodimeric cross-linking galectin-1 and monomeric chimeric galectin-3, with its collagenase-sensitive stalk linked to the carbohydrate-recognition domain, were investigated. Cell-surface expression of the two galectins and accessible galectin-binding sites on various tumor cell lines was ascertained by FACScan analysis. In particular, ligand accessibility for the two galectins differed for the tested cell line types. Binding of tumor cells to laminin and plasma or placental fibronectin was generally reduced by treatment of cells or matrix with galectins. Galectin-3 was more efficient than galectin 1 at impairing laminin's potency as matrix. Cell binding of galectin-1, on the other hand, proved on average more effective for blocking cell association to fibronectins after its preincubation with cell suspensions. Differences were also apparent in the biodistribution of the galectins, where an avian homolog of galectin- served as the control to distinguish effects of spatial and sugar-binding features. Histopathological analysis of lymph-node-negative and -positive breast and colorectal carcinomas (n = 180 including 60 metastatic lesions) indicated a correlation of either increased galectin-1 binding and reduced galectin-3 expression or reduced binding of both galectins with the occurrence of lymph node lesions. Together with data on the heparin-binding lectin, revealing reduced expression to be associated with a positive lymph-node status in the breast cancer group, these results can be interpreted to reflect cell-type-dependent requirements of galectin ligand presentation during the metastatic cascade. By introducing mammalian lectins to lectin-histochemical studies, the detection of quantitative differences in glycosylation brings an understanding of its cell biological significance one step closer.

Adenocarcinoma↗
PubMedDate not supplied

Isolation of immunogenic and lethal peptides of alpha-toxin from Clostridium novyi type B.

The lethal alpha-toxin was isolated from the culture filtrate of Clostridium novyi type B using ammonium sulfate precipitation and ion exchange chromatography. The alpha-toxin has a mol. wt of 190,000 and does not contain any disulfide cross-linkages. It consists of a single polypeptide chain. The peptide fragments resulting from the cyanogen-bromide cleavage were isolated using reversed phase and gel filtration HPLC. The immunogenic actions of these peptides and peptide mixtures were studied in Balb/c mice. Three polyclonal antisera recognizing the uncleaved native toxin could be found using an ELISA test (Br3, Bro2, Bro5). One peptide mixture (Tx5), which was proved lethal in shell-less quail eggs (in vitro), was rechromatographed with gel filtration HPLC that resulted in one peptide with mol. wt 3000 (Txleth), which again proved lethal in the shell-less quail egg lethality test. The immunogenic peptides differ from the lethal one, therefore we assumed different locations on the polypeptide chain. The separation of the immunogenic, non-toxic fragment from the lethal one may allow the production of a highly specific non-toxic vaccine. By using synthetically produced immunogenic peptides, time-consuming purification methods and working with the whole toxin will become unnecessary.

Amino Acid Sequence↗
PubMedDate not supplied

Toxic effects of cyclophosphamide and methylnitrosourea in Japanese quail embryos depend on the route of administration.

We studied the toxic effects of cyclophosphamide and methyl-N-nitrosourea in cultured quail embryos when injected into the albumen and the subembryonic liquid, respectively. The data indicate that both agents display a greater toxicity when given into the subembryonic liquid instead of the albumen. This differential toxicity was demonstrated by means of the survival rate, developmental parameters like growth rate and developmental stages, and excreted nucleosides and uric acid. It is reasonable to assume that the lesser toxicity in the albumen is mainly due to the lower concentration reaching the embryo after the dilution of the agent in this compartment. On the basis of the results obtained in these experiments, the cultured avian embryo represents an especially suitable experimental system to study the changes in metabolism induced by toxic agents. This is due to the easy access to the subembryonic compartment and the easy sampling of allantoic liquid under these conditions.

Allantois↗
PubMedDate not supplied

Expression of endogenous lectins (galectins, receptors for ABH-epitopes) and the MIB-1 antigen in esophageal carcinomas and their syntactic structure analysis in relation to post-surgical tumor stage and lymph node involvement.

Squamous cell carcinomas of the esophagus, a disease with poor prognosis, are especially frequent in China and South Africa. To initiate the study of endogenous lectins in this tumor class we employed synthetic neoglycoconjugates and focused on galectins as markers. Histological sections of 43 cases of esophageal carcinomas were analyzed with labeled galectins-1 and -3 and their specific antibodies, neoglycoconjugates exposing chemically prepared histo-blood group A-, B- and H-trisaccharides and the antibody MIB-1 (Ki-67). Features of structural and numerical staining intensities determined quantitatively were correlated to clinical data sets of pTN stages, sex and age of patients. Low tumor stages (pT1/T2) were seen in 10/43 cases (23%) and 65% of the carcinomas surgically treated lacked notable lymph node involvement (pN0). The women were younger than the men (47 years versus 54 years). The proliferation activity of the tumor cells was high and amounted to 75% at average. The presence of galectin-1 and the structural entropy of distribution of staining with carrier-immobilized A-trisaccharide were associated with pN stages. These initial data indicate that distinct glycohistochemical features appear to have prognostic significance in this tumor class, adding to the emerging significance of this marker class in lung cancer.

ABO Blood-Group System↗