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Biomedical subjects

W Breuer

Publications and source records attributed to W Breuer.

At least 37 records · Page 2Linked to original sources
PubMed1999

Myelodysplastic syndrome (MDS), acute myeloid leukaemia (AML) and chronic myeloproliferative disorder (CMPD) in cats.

Histological, enzyme histochemical and ultrastructural findings in three cases of feline bone marrow neoplasia are described. The following changes were observed: in myelodysplastic syndrome (MDS), a low medullary blast count, strongly atypical (micromegakaryocytic) proliferative megakaryocytopoiesis, hypoplastic erythrocytopoiesis with impairment of differentiation, multifocal extravasation and lymphoid aggregates; in acute myeloid leukaemia (AML), medullary proliferation of undifferentiated cell types; in chronic myeloIproliferative disorder (CMPD), trilinear medullary proliferation with complete cellular maturation, osteomyelosclerosis and extramedullary haemopoiesis. In two cases (MDS, AML), ultrastructural demonstration of C-type virus particles (feline leukaemia virus) suggested a viral aetiology.

Acute Disease↗
PubMed1999

Prognostic value of histopathological grading in canine extramedullary plasmacytomas.

The canine extramedullary plasmacytoma (cEMP) has recently been the subject of numerous investigations, indicating that the histomorphologic diagnosis is often difficult because of the variety of morphologic features. Therefore, the objective of this study was to establish a subclassification scheme for cEMPs and to evaluate correlations between the types and malignancy. Retrospectively, 117 cEMPs, all immunohistochemically characterized by a monoclonal immunoglobulin light-chain expression, were collected and assigned to morphologic types. These types were compared using data from a follow-up study on metastasis and tumor recurrence, then compared by proliferation rate, determined by immunohistochemical detection of the antigen Ki-67. Histopathologic typing revealed five different types of cEMPs, ranging from the mature type with typical plasma cells to the polymorphous-blastic type. Between these two forms, three additional types were established: hyaline, cleaved, and asynchronous. Most of the cEMPs were of the cleaved and asynchronous types. In all cEMPs, mononuclear and multinuclear giant cells were present to varying degrees. Although the results of cell proliferation and the follow-up study indicated less benign behavior by the polymorphous-blastic type, the proliferation rate revealed no statistically significant differences among the cEMP types. The clinical data therefore confirmed previous findings that the risk of tumor recurrence and metastasis in general can be classed as low. The established cEMP typing system is probably a very helpful diagnostic tool, although the types cannot be used for a tumor grading system.

Animals↗
PubMed1998

Bone-marrow changes in infectious diseases and lymphohaemopoietic neoplasias in dogs and cats--a retrospective study.

Bone-marrow changes in infectious diseases due to feline infectious peritonitis virus (FIPV), feline immunodeficiency virus (FIV), parvovirus (PV, canine and feline) and canine distemper virus (CDV), and in the lymphohaemopoietic neoplasias (LHNs) usually associated with feline leukaemia virus infection were studied in samples obtained from 204 cats and 82 dogs at necropsy. The study demonstrated (1) no changes, (2) non-specific reactive changes, and (3) disease-specific changes (similar to those occurring in extramedullary sites) in: 51.2, 48.8 and 9.7% of 41 cases of FIPV infection, respectively; 0, 100 and 0% of nine cases of FIV infection, respectively; 1.3, 0 and 92% of 75 cases of canine PV infection, respectively; 5.3, 3.9 and 84% of 76 cases of feline PV infection, respectively; 71.4, 28.6 and 0% of seven cases of CDV infection, respectively; and 35.9, 52.6 and 11.5% of 78 cases of LHN, respectively. The distribution of the disease-specific bone-marrow changes was either diffuse or focal; diffuse changes were frequently found in cases of feline and canine PV infection, and focal changes were found inconsistently in FIPV infections and feline LHN. To the extent that the bone marrow showed any changes in FIV and CDV infections, they were mostly reactive and not pathognomonic.

Animals↗
PubMed1998

Large granular lymphocyte leukemia/lymphoma in six cats.

This report describes six cases of feline large granular lymphocyte lymphoma identified by light microscopy on the basis of their characteristic azurophilic granulation in Giemsa-stained plastic sections and by electron microscopy on the basis of their typical granules. Although the granules of all the tumor cells were negative for peroxidase activity, they all demonstrated chloroacetate-esterase and acid phosphatase activity. All the tumors reacted with cross-reacting antibodies against the CD3 antigen (epsilon chain) and did not react with a cross-reacting monoclonal antibody directed against epitopes on cytoplasmic domains of the CD20 antigen. Three tumors had a positive reaction with a monoclonal human CD57-like antibody. This is highly suggestive of either a cytotoxic T cell or a natural killer cell origin of the neoplasias. In three cats, although other abdominal organs were affected to a variable extent, the main neoplastic lesions were localized in the gastrointestinal tract and the jejunal lymph nodes. In contrast, in the other three cats, organ involvement was more widespread, affecting the lung (two), myocardium (two), precardiac mediastinum (one), salivary gland (one), and spinal cord (one); in addition, leukemia was present in two of these cats. The data presented indicate that tumors made up of large granular lymphocytes occur more frequently in cats than previously assumed and that they share many characteristic features with specific subtypes of clonal disorders of large granular lymphocytes in humans.

Acid Phosphatase↗
PubMed1998

[Diabetes mellitus in young dogs and cats. Case reports and a comparison with relevant literature].

Cases of diabetes mellitus in young cats and dogs (younger than five years) are described rarely in contrary to reports in older patients (older than eight years). Histological alterations of pancreatic islets in young canine and feline diabetic patients represent the main issue of this publication together with the regenerative capacity of insulin-producing cells in these patients. These regenerative phenomenons of differentiated islet cells (replication) or their progenitors (neogenesis deriving from stem cells) are detected in young cats and dogs in isolated cases whereas they are in general missing in older patients. In this publication, own cases are presented together with histopathological as well as clinical findings. The cases are discussed with the relevant literature of veterinary and human medicine.

Age Factors↗
PubMed1997

Fluorescence analysis of the labile iron pool of mammalian cells.

The labile iron pool (LIP) of cells constitutes a cytosolic fraction of iron which is accessible to permeant chelators and contains the cells' metabolically and catalytically reactive iron. LIP is maintained by a balanced movement of iron from extra- and intracellular sources. We describe here an approach for tracing LIP levels in living cells based on the fluorescent probe calcein (CA). This probe binds Fe(II) rapidly, stoichiometrically, and reversibly while forming fluorescence-quenched CA-Fe complexes. Cells are loaded with CA via its acetomethoxy precursor CA-AM, attaining 1-10 microM intracellular concentrations and retaining full viability. LIP is defined here operationally as the sum of "free" and CA-bound iron of the cell. The method for assessing LIP is based on the measurement of: (a) the total intracellular concentration of CA in CA-loaded cells ([CA]1), which is estimated from fluorimetric measurements of CA in a given suspension of cells, the number of cells, and the cell volume; (b) the intracellular [CA-Fe], the concentration of [CA] bound to metals (> 95% iron), which is assessed from the relative rise in fluorescence (delta F) elicited by addition of highly permeant and high-affinity binding chelators such as salicyladehyde-isonicotinoyl-hydrazone (SIH) and the value of [CA]1; and (c) the "free" cell iron concentration [Fe(II)], which is computed from the experimentally determined values of CA-Fe(II)'s dissociation constant (Kd) in various cell lines grown in suspension (Kd = 0.22 +/- 0.01 microM). The value of cellular LIP is defined as the sum of [CA-Fe] and [Fe]. It is derived from the experimental determination of [CA]1 and [CA-Fe] and from calculation of [Fe] by application of the mass law equation using the Kd value of [CA-Fe]. The estimated values of LIP for resting erythroid and myeloid cells are in the range of 0.2-1.5 microM. The values varied commensurately with cell iron loads and iron chelator treatment. The method provides a simple, noninvasive tool for on-line monitoring of cytosolic iron under normal and abnormal conditions of cell iron supply and for assessing the dynamics of intracellular iron in living cells.

Animals↗
PubMed1997

Newly delivered transferrin iron and oxidative cell injury.

Cell iron status was assessed in terms of its capacity to mediate cell injury by pro-oxidants. Cultured K562 cells, which maintain a stable cytosolic labile iron pool (LIP) of < 0.5 microM, underwent distinct changes after short exposures to transferrin (Tf) followed by t-butyl hydroperoxide (TBHP): (a) rise in LIP, detectable fluorimetrically; (b) increased lipid peroxidation and (c) eventual cell death. All of these effects were inhibited by weak bases or iron chelators. Similarly, hydrogen peroxide caused rises in both LIP and oxidant species detectable with 2',7'-dichlorofluorescin diacetate, which were enhanced by preincubation with Tf. The Tf-delivered iron disappeared from LIP and the TBHP-reactive pool with a t1/2 < 30 min. The results indicate that the catalytic potential of iron is highest while in transit between endosomes and cytosolic ligands.

Cell Survival↗
PubMed1997

Epoxyethylglycyl peptides as inhibitors of oligosaccharyltransferase: double-labelling of the active site.

Pig liver oligosaccharyltransferase (OST) is inactivated irreversibly by a hexapeptide in which threonine has been substituted by epoxyethylglycine in the Asn-Xaa-Thr glycosylation triplet. Incubation of the enzyme in the presence of Dol-PP-linked [14C]oligosaccharides and the N-3,5-dinitrobenzoylated epoxy derivative leads to the double-labelling of two subunits (48 and 66 kDa) of the oligomeric OST complex, both of which are involved in the catalytic activity. Labelling of both subunits was blocked competitively by the acceptor peptide N-benzoyl-Asu-Gly-Thr-NHCH3 and by the OST inhibitor N-benzoyl-alpha,gamma-diaminobutyric acid-Gly-Thr-NHCH3, but not by an analogue derived from the epoxy-inhibitor by replacing asparagine with glutamine. Our data clearly show that double-labelling is an active-site-directed modification, involving inhibitor glycosylation at asparagine and covalent attachment of the glycosylated inhibitor, via the epoxy group, to the enzyme. Double-labelling of OST can occur as the result of either a consecutive or a syn-catalytic reaction sequence. The latter mechanism, during the course of which OST catalyses its own 'suicide' inactivation, is more likely, as suggested by indirect experimental evidence. The syn-catalytic mechanism corresponds with our current view of the functional role of the acceptor site Thr/Ser acting as a hydrogen-bond acceptor, not a donor, during transglycosylation.

Animals↗
PubMed1997

Identification of lambda light chain amyloid in eight canine and two feline extramedullary plasmacytomas.

Amyloid deposition in varying amounts and with variable patterns of distribution (focal or diffuse) was demonstrated in eight canine and two feline extramedullary plasmacytomas expressing lambda light chains. Frequently, the neoplastic plasma cells had been displaced by the amyloid deposits. Foreign-body giant cells were regularly detected in the vicinity of the amyloid. In all 10 cases, Congo-red staining of the amyloid was resistant to potassium permanganate oxidation. Immunohistochemically, the amyloid reacted positively with cross-reacting antibodies against human and equine A lambda amyloids. Extramedullary plasmacytomas accompanied by localized AL amyloidosis have so far been described in human beings, dogs, cats and horses.

Amyloidosis↗
PubMed1997

Chelation and mobilization of cellular iron by different classes of chelators.

Iron chelators belonging to three distinct chemical families were assessed in terms of their physicochemical properties and the kinetics of iron chelation in solution and in two biological systems. Several hydroxypyridinones, reversed siderophores, and desferrioxamine derivatives were selected to cover agents with different iron-binding stoichiometry and geometry and a wide range of lipophilicity, as determined by the octanol-water partition coefficients. The selection also included highly lipophilic chelators with potentially cell-cleavable ester groups that can serve as precursors of hydrophilic and membrane-impermeant chelators. Iron binding was determined by the chelator capacity for restoring the fluorescence of iron-quenched calcein (CA), a dynamic fluorescent metallosensor. The iron-scavenging properties of the chelators were assessed under three different conditions: (a) in solution, by mixing iron salts with free CA; (b) in resealed red cell ghosts, by encapsulation of CA followed by loading with iron; and (c) in human erythroleukemia K562 cells, by loading with the permeant CA-acetomethoxy ester, in situ formation of free CA, and binding of cytosolic labile iron. The time-dependent recovery of fluorescence in the presence of a given chelator provided a continuous measure for the capacity of the chelator to access the iron/CA-containing compartment. The resulting rate constants of fluorescence recovery indicated that chelation in solution was comparable for the members of each family of chelators, whereas chelation in either biological system was largely dictated by the lipophilicity of the free chelator. For example, desferrioxamine was among the fastest and most efficient iron scavengers in solution but was essentially ineffective in either biological system when used at < or = 200 microM over a 2-hr period at 37 degrees. On the other hand, the highly lipophilic and potentially cell-cleavable hydroxypyridinones and reversed siderophores were highly efficient in all biological systems tested. It is implied that in K562 cells, hydrolysis of these chelators is relatively slower than their ingress and binding of intracellular iron. The chelator-mediated translocation of iron from cells to medium was assessed in 55Fe-transferrin-loaded K562 cells. The speed of iron mobilization by members of the three families of chelators correlated with the lipophilicity of the free ligand or the iron-complexed chelator. The acquired information is of relevance for the design of chelators with improved biological performance.

Fluorescent Dyes↗
PubMed1997

Canine neuroendocrine tumors of the pancreas: a study using image analysis techniques for the discrimination of metastatic versus nonmetastatic tumors.

Canine pancreatic neuroendocrine tumors were studied using different image analysis techniques (nuclear image histometry, analysis of argyrophilic proteins of nucleolar organizer regions, determination of the mouse anti-Ki 67 antigen proliferation index, and DNA densitometry) to correlate their biological behavior with objective phenotypic markers. The methods were compared to determine the best method for distinguishing between metastatic and nonmetastatic tumors. Discrimination between the two types of tumor was possible using nuclear image histometry in combination with morphometric analysis of argyrophilic proteins of nucleolar organizer regions. In contrast, the mouse anti-Ki 67 antigen proliferation index, DNA measurement, and immunohistochemical parameters revealed no significant difference between the two types of tumors.

Animals↗
PubMed1997

Ductuloendocrine cell proliferation in the pancreas of two young dogs with diabetes mellitus.

Two cases of diabetes mellitus in juvenile dogs (a 3-4 month-old Golden Retriever and a 2-month-old Labrador Retriever) are described here in terms of their clinical, histologic, and immunohistologic findings. Only very few insulin-positive cells were demonstrated immunohistochemically in one dog. In the second dog, the alterations of the pancreas consisted of hydropic vacuolar degeneration of B cells in the islets of Langerhans. In both cases, hyperplasia of the vacuolated cells was prominent. These cells formed tubular structures and were immunohistochemically positive for cytokeratin and proliferating cell antigen (MIB-1). Furthermore, within these vacuolated areas, some cells were positive to varying degrees for insulin, glucagon, somatostatin, and pancreatic polypeptide. Apoptotic cells could be seen in the exocrine pancreas, in vacuolated areas, and occasionally in the islets of both dogs. We interpret these alterations as ductuloendocrine cell proliferation, probably as an idiopathic compensatory response.

Age Factors↗
PubMed1997

[Adenovirus-induced pneumonia in two guinea pigs].

Two cases of a rare virus-induced disease of Guinea pigs are described. The animals had died because of a pneumonia. Large intranuclear viral inclusion bodies could be demonstrated in the endothelial cells of the lungs. In the second case inclusion bodies could be found additionally in macrophages of the spleen and in macrophages, hepatocytes and endothelial cells of the liver. By means of electron microscopy the virus could be specified as adenovirus.

Adenoviridae Infections↗
PubMed1997

[Porcine proliferative enteropathy in feeders and breeder sows].

Porcine proliferative enteropathy (PPE) was recognized as an important economic factor in pig production. In the following, three cases of PPE in pigs presented for necropsy are discussed. The animals were raised in a stud stock and a weaning production facility in Bavaria. The clinical signs described in the history indicated a herd problem with hemorrhagic diarrhoea and paleness being the cardinal signs. The diagnosis was based upon gross and histopathological findings and was confirmed by transmission electron microscopy. The different types of manifestation of the disease seen in the two production facilities are discussed in the context of the current literature on etiology and pathogenesis.

Animals↗
PubMed1996

Differential cytotoxicity of iron chelators on malaria-infected cells versus mammalian cells.

Iron chelators of the hydroxamate class arrest in vitro proliferation of malaria parasites end of mammalian cells. The factors determining the biological activity of the chelators have classically been attributed to the chelators' capacity for binding iron and to their ability to traverse membranes as free chelators and as chelator-iron complexes. We show in this work that the nature of the chelatable pool of cell iron also contributes to the susceptibility of cells to iron chelators. A class of N-terminal (Nt derivatives of desferrioxamine (DFO), (Nt-DFO), is shown here to differentially affect growth and replication of intraerythrocytic parasites (Plasmodium falciparum). Methyl-anthranilic DFO (MADFO), the relatively less hydrophilic member of the Nt-DFOs series, reduced parasite proliferation (48 hour test) with an IC50 of 4 +/- 1 micromol/L and mammalian cell (K562 and HepG2) proliferation with an IC50 > 100 micromol/L. On the other hand, the more hydrophilic Nt-free DFO, displayed IC50 values of 21 +/- 5 micromol/L for parasites and 7 +/- 1 micromol/L for mammalian cells. The selective antiparasitic activity of MA-DFO, as reflected in the speed of action and IC50 values on cell proliferation, is attributed primarily to membrane permeation and iron (III) binding properties of the drug. In contrast, the relatively low antiproliferative activity of the more permeant MA-DFO on mammalian cells, resulted from MA-DFO's reduced capacity for scavenging intracellular iron. This is apparent from MA-DFO reduced effects on: (1) the chelatable iron (II) pool that is associated with the cell cytosol; (2) the cell chelator-extractable iron, and (3) cell ferritin levels. The potent antimalarial efficacy and biological selectivity of MA-DFO relative to the parent DFO, is of importance for improved design of chemotherapeutic agents.

Animals↗
PubMed1996

Dynamics of the cytosolic chelatable iron pool of K562 cells.

The labile iron pool of cells (LIP) constitutes the primary source of metabolic and catalytically reactive iron in the cytosol. We studied LIP homeostasis in K562 cells using the fluorescent metal-sensitive probe calcein. Following brief exposure to iron(II) salts or to oxidative or reductive stress, LIP rose by up to 120% relative to the normal level of 350nM. However, the rate of recovery to normal LIP level differed markedly with each treatment (respective t1/2s of 27, 65-88 and < or = 17 min). We show that the capacity of K562 cells to adjust LIP levels is highly dependent on the origin of the LIP increase and on the pre-existing cellular iron status.

Cytosol↗
PubMed1996

A fluorescence assay for assessing chelation of intracellular iron in a membrane model system and in mammalian cells.

Iron chelators are important tools in biochemical studies of iron metabolism and in the therapy of iron overload diseases. Their mode of action is comprised of entry into cells and scavenging intracellular metal, which includes complexation and egress of the complex. Iron is a metal which appears in the cells in various chemical forms and in different compartments. The form of the metal directly affected by the chelators is the most labile, low-molecular-weight type, which is present in the cytosol and is known as the "chelatable iron." This form is thought to be in dynamic equilibrium with several sequestered forms present in the cell, including the iron-responsive proteins. We recently introduced a fluorescent method for assessing the chelatable iron pool of cells, based on the quenching of the fluorescent calcein by metal ions (Breuer et al., J. Biol. Chem., 1995, 270, 24209-24215). In this work we adapted the method for dynamic assessment of chelator efficacy in scavenging iron from cells. In assay 1, red blood cells ghosts are used as a cell membrane model. The free-acid (impermeant) form of calcein is loaded into ghosts by encapsulation (lysis and resealing) and its fluorescence is quenched by addition of permeant iron(II). Chelators added to ghosts lead to iron removal from calcein and hence to recovery of fluorescence, commensurate with their permeation into ghosts and iron binding affinity. In assay 2, human K562 erythroleukemia cells are loaded with calcein via its permeating and cleavable acetoxymethyl form. A fraction of the intracellular calcein fluorescence is quenched in situ by endogenous cellular iron. The rate of dequenching which is obtained after addition of a chelator provides a measure for the scavenging of the intracellular metal, a process which, as in assay 1, depends on chelator permeation and binding affinity for iron. The two methods provide convenient means for assessing the efficacy of candidate chelator structures in depleting cell iron pools. They are also potentially applicable to chelation of other metals such as Co(II), Ni(II), and Cu(II) and to any cellular system or membrane vesicles.

Erythrocyte Membrane↗