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W Eichler

Publications and source records attributed to W Eichler.

At least 37 records · Page 2Linked to original sources

Perioperative tissue thickness measurement by a new miniature ultrasound device.

INTRODUCTION: A recently developed mini ultrasound device for measurement of peripheral tissue thickness is now available for use in clinical practice. Whether this device allows a better guidance of perioperative fluid therapy has to be investigated. Therefore, it is necessary to get basic data on the parameter tissue thickness in otherwise healthy patients during surgery. The aim of the present study was to evaluate differences in tissue thickness change between patients in supine and head down position with a novel handheld ultrasound device during the perioperative course of healthy surgical patients under a standardized fluid regimen. METHODS: After obtaining ethics committee approval and informed consent we studied 19 ASA 1-2 female patients undergoing gynecological procedures in supine (SUP, n = 11) or in 30 degrees head down position (HD, n = 8) in general anesthesia. Preoperative NPO status was comparable in both groups. Lactated Ringer's solution (LR) was continuously infused at a rate of 8 ml/kg b.w./h over 90 min and tissue thickness (TT) was determined by ultrasound before induction (t0) and in 30 min intervals (t30, t60, t90) at the forehead. Simultaneously plasma viscosity (PV) was evaluated. RESULTS: Group SUP presented at t0 a forehead TT of 5.3 mm (SD +/- 0.5), at t30 TT was unchanged. At t60 mean TT increased significantly to 5.6 mm, (+/- 0.6). At t90 mean TT remained stable at 5.7 mm (+/- 0.5). Group HD presented at t0 a mean TT of 4.6 mm (+/- 0.7), at t30 mean TT was 4.9 mm (+/- 0.7) and at t60 mean TT of the forehead skin was measured as 5.3 mm (+/- 0.6). Significance to t0 was reached at t90 with a mean TT of 5.4 mm (+/- 0.7). Group HD showed a steeper increase and a parallel stabilization phase at the end. Differences between t0 and t90 have been significant. Mean PV in the SUP group at t0 (1.361 mPa*s, SD: +/- 0.045) decreased under the infusion therapy to 1.276 mPa*s (+/- 0.04) at t90. Mean PV in the HD group was determined 1.351 mPa*s (+/- 0.06) at t0 and declined to 1.274 mPa*s (+/- 0.03) at t90. CONCLUSIONS: The findings suggest that fluid replacement after an NPO period and the expected changes of forehead TT due to positioning of the patient are detectable by this new ultrasound device.

Adult↗

CD97: a dedifferentiation marker in human thyroid carcinomas.

CD97 is a dimeric glycoprotein of Mr 75,000-85,000 and 28,000 belonging to a novel subfamily of seven-span transmembrane region leukocyte cell surface molecules. It is expressed abundantly in cells of hematopoietic origin. This is the first report demonstrating the expression of CD97 outside the hematopoetic system. CD97 was studied in normal human and neoplastic follicular epithelium of the thyroid and anaplastic (n = 3) and papillary (n = 1) thyroid carcinoma cell lines. In normal thyroid tissue (n = 11), no immunoreactivity of CD97 could be found, whereas in differentiated thyroid carcinomas (n = 10), CD97 expression was either lacking or low. Eleven of 12 undifferentiated anaplastic carcinomas revealed high CD97 presentation. CD97 was absent or only weakly present in patients with postoperative T1 tumors but increased greatly with the progression to postoperative T4 tumors. CD97 is clearly present in thyroid carcinoma cell lines but only at a very low level in normal human thyrocytes. Quantitation of CD97 cell surface expression levels revealed that C 643 and SW 1736 cells showed a two to four times higher specific antibody-binding capacity than did 8505 C and HTh 74 cells and a nearly 20 times higher specific antibody-binding capacity than normal thyrocytes. Phorbol 12-myristate 13-acetate treatment progressively caused a decrease of CD97 antigen expression in all cell lines to about 30% of their initial levels after 48 h. Immunohistochemical staining of SW 1736 cells revealed that CD97 is located in most of the cell compartments and suggested a CD97 internalization process after phorbol 12-myristate 13-acetate treatment. Semiquantitative reverse transcription-PCR showed a correlation of CD97 mRNA and cell surface CD97 expression level in the cell lines. SW 1736, HTh 74, and 8505 C cells apparently expressed CD97 with alternative glycosylation compared to peripheral lymphocytes, whereas most of the CD97 antigen presented on thyrocytes and C 643 cells had glycosylation sites resembling those of lymphocytes. The data suggest that CD97 expression may be a sensitive marker of dedifferentiation and of lymph node involvement in human thyroid tumors.

Antibodies, Monoclonal↗

Expression characteristics of the human CD97 antigen.

Molecules whose expression is increased upon stimulation on leukocyte populations are of considerable interest because insights into their structure and function extend knowledge of the intracellular and intercellular events that accompany cellular activation. One such molecule is CD97, a cell surface antigen that is rapidly upregulated upon activation on lymphocytes. Increasing CD97 levels on peripheral blood lymphocytes (PBL) induced by different stimuli were found to be largely independent on de novo RNA and protein synthesis during the early stage of activation. Thus, inhibition of CD97 surface expression by cycloheximide was not noticeable or insignificant for 15 min to 4 h after stimulation. Furthermore, a fraction of intracellular CD97 decreased within 2 h suggesting redistribution of CD97 protein. Later, de novo protein synthesis apparently contributes to the induction of high CD97 surface density and inhibition by cycloheximide was more pronounced. Upregulation of CD97 on PBL involves protein kinase C-, tyrosine protein kinase- and Ca2+-dependent intracellular pathways. The effect on CD97 surface expression of the phorbol ester, phorbol 12-myristate 13-acetate (PMA), is different in PBL and Jurkat T cells. Whereas it stimulated after 22 h strong CD97 increase on PBL it suppressed CD97 expression on Jurkat T cells. CD97 expression, which is strong and constitutive on myelo-monocytic cells, is shown to be not leukocyte-restricted. Thus, CD97 transcripts were found in most of the investigated nonhematopoietic cell types and CD97 protein was detected on the cell surface at low amount. In a previous report, sequence data of a CD97 cDNA suggested a protein homologous to the secretin receptor superfamily and consisting of 722 amino acids with 8 potential glycosylation sites. According to this finding, glycoproteins displaying apparent molecular weights in the region of 70-85 kDa were detected in all investigated cell types. These molecules may represent differentially glycosylated and sialylated molecular forms of the same polypeptide. The findings support the notion that CD97 is broadly distributed and possesses a differentially regulated expression behavior on leukocytes and non-hematopoietic cells.

Antigens, CD↗

Sevenspan transmembrane molecules: novel receptors involved in leukocyte adhesion.

CD97 is a member of a new subgroup of seven-span transmembrane (7-TM) molecules which belong to the secretin receptor superfamily. Different from other members of the secretin receptor family, these recently characterized molecules have extended extracellular regions comprising several EGF domains near the NH2 terminus. We recently demonstrated that the extracellular part of CD97 is involved in intercellular adhesion since it specifically binds to CD55 (decay accelerating factor), a regulatory protein of the complement cascade. To our knowledge this is the first demonstration of a cellular ligand for a 7-TM molecule.

Antigens, CD↗

Conformational changes in CD45 upon monoclonal antibody crosslinking.

The CD45 protein tyrosine phosphatase is expressed in different isoforms that result from alternative splicing of three exons (A, B, and C) encoding regions near the N-terminus of the extracellular part of the molecule. We describe here a novel epitope on the N-terminal end of CD45 that is recognized by the MAb BL-TSub/2. Crossblocking studies showed that BL-TSub/2 and UCHL1 (CD45RO) binding sites are partially overlapping. However, in marked contrast to the CD45RO epitope, protease treatment of cells strongly diminished BL-TSub/2 binding. Similar to the UCHL1 epitope, the BL-TSub/2 binding site involves carbohydrate moieties, since neuraminidase treatment abrogated the reactivity of the MAb. Markedly, preincubation of cells with both CD45 common and CD45RA MAb induced a pronounced increase of BL-TSub/2 binding. This latter finding suggests that crosslinking of the CD45 molecule leads to conformational changes that could influence association of the molecule with putative ligands.

Animals↗

Expression cloning and chromosomal mapping of the leukocyte activation antigen CD97, a new seven-span transmembrane molecule of the secretion receptor superfamily with an unusual extracellular domain.

CD97 is a monomeric glycoprotein of 75 to 85 kDa that is induced rapidly on the surface of most leukocytes upon activation. We herein report the isolation of a cDNA encoding human CD97 by expression cloning in COS cells. The 3-kb cDNA clone encodes a mature polypeptide chain of 722 amino acids with a predicted molecular mass of 79 kDa. Within the C-terminal part of the protein, a region with seven hydrophobic segments was identified, suggesting that CD97 is a seven-span transmembrane molecule. Sequence comparison indicates that CD97 is the first leukocyte Ag in a recently described superfamily that includes the receptors for secretin, calcitonin, and other mammalian and insect peptide hormones. Different from these receptors, CD97 has an extended extracellular region of 433 amino acids that possesses three N-terminal epidermal growth factor-like domains, two of them with a calcium-binding site, and a single Arg-Gly-Asp (RGD) motif. The existence of structural elements characteristic for extracellular matrix proteins in a seven-span transmembrane molecule makes CD97 a receptor potentially involved in both adhesion and signaling processes early after leukocyte activation. The gene encoding CD97 is localized on chromosome 19 (19p13.12-13.2).

Amino Acid Sequence↗

Characterization of an early activation-dependent antigen on lymphocytes defined by the monoclonal antibody BL-Ac(F2).

A novel activation-dependent lymphocyte cell-surface antigen which is recognized by a MoAb, BL-Ac(F2), is described. Although not found on resting lymphocytes the antigen is induced rapidly within 2-4 h following stimulation of the cells using mitogens or antibodies against the T-cell CD3 antigen and sIgM on B cells, respectively. Immunoprecipitation and Western Blotting indicated that the MoAb recognizes a molecule in a range of 78-85 kDa. Beyond its activation-dependent expression on lymphocytes the antigen was detected also on myelo-monocytic cells. Expression kinetics and cellular distribution of this molecule suggest that it is distinct from previously described activation-dependent cell-surface antigens such as CD69, CD25 and 4F2.

Animals↗

Characterization of DNA-directed RNA polymerases in isolated macronuclei of the ciliated protozoan Tetrahymena thermophila. Effects of purified ornithine decarboxylase and amine compounds.

The possible regulatory interactions of purified ornithine decarboxylase with DNA-directed RNA polymerases in isolated macronuclei from the ciliated protozoan Tetrahymena thermophila were studied. It has been found that highly purified ODC (specific activity 10.2 mumols CO2 x h-1 x mg-1), even at activities of 37,500 nmol CO2 x h-1 per ml failed to alter RNA polymerase activity in the in vitro transcription assay in the presence or absence of the substrate L-ornithine at 20mM. The naturally occurring di- and polyamines putrescine, spermidine, and spermine stimulated in-vitro-transcription in isolated macronuclei more at optimal Mg2+/Mn(2+)-concentrations than at suboptimal concentrations, suggesting that polyamines act via a mechanism which is distinct from that of the inorganic cations. Of the monovalent amine compounds tested, (NH4)+ at high concentrations between 40 and 50mM slightly stimulated activity whereas the onset of stimulation by the organic amine compounds, piperidine and cyclohexylamine, was inversely related to the hydrophobicity of each particular compound. In the series of divalent amines, the correct distance between the N-atoms appeared to be very important since ethylenediamine and piperazine did not stimulate significantly but did inhibit at concentrations above 5 mM. 1,3-Diaminopropane stimulated slightly but inhibited above 10 mM, whereas the 1,4-diamino compounds putrescine and 1,4-diaminocyclohexane (DAC) were equally potent stimulators with the more hydrophobic one, DAC, reaching the maximum at lower concentrations than putrescine. For the trivalent amines, the influence of correct spacing seems not to be as important: N-(2-aminoethyl)piperazine stimulated very similar to spermidine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Polyamine biosynthesis in arginine-starved and refed rats.

Growth of rats fed with a synthetic diet was studied under control conditions (arginine-rich), arginine starvation, and arginine starvation/refeeding. Hepatic polyamine concentrations and ornithine decarboxylase (ODC-)activity were determined for each population. In the livers of arginine-starved rats putrescine was decreased to half the control content within 8 days; upon refeeding, it returned to control levels within another 8 days. Spermidine content in liver tissue of arginine-starved rats remained rather stable for 7 days, but thereafter dropped to half the original value within two days. Refeeding for a period of 11 days was not enough to restore the spermidine content. The effects of arginine starvation/refeeding on spermine were very similar to those of spermidine. ODC specific activity, when correlated with growth, was higher in livers of arginine-starved rats than in control animals. Refeeding caused a decrease in ODC-activity although growth arrest was completely released. This apparent uncoupling of growth and ODC stimulation supports the theory that ODC in rat liver is regulated at three levels: first the growth-related component which is observed after stimulation by growth-hormone; second the known feed back control by polyamines, e.g. via antizyme; third the regulation at the level of the substrate supply which has been shown in this work. This is not a unique finding since very similar results have been obtained in previous experiments with the protozoan Tetrahymena thermophila. A remarkable observation of these assays was that L-ornithine, when added to the arginine-free diet was not able to substitute for L-arginine in directing growth and growth related processes.

Animals↗

[Characterization of a T-cell subpopulation with a new monoclonal antibody BL-TSub/2].

The mab BL-TSub/2 binds to a majority (80%) of thymocytes. The antigen isolated from these cells has an apparent molecular weight of about 170 kDa. The mab BL-TSub/2 recognized antigen is expressed on approximately 50% of mature T cells in peripheral blood. The mab BL-TSub/2 does not bind to monocytes, granulocytes, platelets and CD16+NK cells. The antigen is possibly present on a part of peripheral B cells in very low density. Double labelling experiments showed that about 30% of CD4+ and 60% of CD8+ cells react with the mab BL-TSub/2. This BL-TSub/2+ subset is not identical with T-cell subpopulations till now defined. Within the CD4+ subset the majority of BL-TSub/2+ cells is included in the CD45RO+ subset while only a small proportion is CD45RA+. However, the CD8+ BL-TSub/2+ cells are mostly CD45RA+ with only few cells belonging to the CD45RO+ subset.

Antibodies, Monoclonal↗

[Epitope analysis of CD68 antigens].

Different epitopes of CD68 antigen are detectable by three new generated monoclonal antibodies (MABs) BL-M68/1-3 (all mouse IgM). The MAB BL-M68/3 reacts with an epitope, which is stable also in formalin-fixed paraffin-embedded tissues. In immunohistochemical blocking experiments using the own biotinylated MABs and six CD68 MABs from the IV. Workshop on Human Leucocyte Differentiation Antigens two new epitopes could be defined. The number of known CD68 epitopes increases at least up to seven.

Antibodies, Monoclonal↗

[Characterization of a monoclonal antibody against a differentiation antigen of human natural killer cells (NK cells)].

The new murine monoclonal antibody BL-(H5) reacts with a novel surface molecule which is mainly expressed on human NK and B cells. The antigen is not expressed on peripheral T lymphocytes, thymocytes and different human T-cell lines. BL-(H5) does not bind to erythrocytes and platelets. The monoclonal antibody reacts in western blotting experiments with an antigen of 78kDa.

Antibodies, Monoclonal↗

Regulation of growth and polyamine biosynthesis of the ciliated protozoan Tetrahymena thermophila. Effects of L-arginine metabolites and polyamines.

Growth of Tetrahymena thermophila in a synthetic nutrient medium with or without the essential amino acid L-arginine was studied in the presence or absence of the arginine metabolites L-citrulline and L-ornithine and the polyamines putrescine, spermidine, and spermine. The effects of the growth conditions on the stimulations of the enzymes of the arginine metabolic and polyamine biosynthetic pathway, arginine deiminase (ADI), citrulline hydrolase (CH), ornithine decarboxylase (ODC), and ornithine-oxo-acid aminotransferase were determined. Tetrahymena cells were unable to grow in the absence of L-arginine and the amino-acid utilization was greatly impaired. None of the metabolites or polyamines was able to substitute for arginine. In the presence of arginine, Tetrahymena cultures grew well and citrulline and ornithine did not alter the growth behaviour in any way. In the presence of putrescine, the lag period was decreased from 3 h to 2 h. Spermidine and spermine acted similar to putrescine but less pronounced. The stimulation of the activity of ADI, the key enzyme of arginine degradation, was absolutely dependent upon the presence of arginine in the medium: in the absence of arginine, the low ADI activity which was present in the cells before inoculation was decreased to zero levels within 30 min. In the presence of arginine, the stimulation of ADI was not altered by citrulline and ornithine but putrescine, spermidine, and spermine decreased ADI-stimulation to half of the control values. The stimulation of CH activity in the presence of arginine was not altered by any added metabolite or polyamine. In the media without arginine, stimulation of CH was greatly reduced, in the presence of ornithine more than in its absence, and even more in the presence of putrescine and spermidine. Stimulation of ODC activity in the presence of arginine was not affected by citrulline and ornithine but in the presence of polyamines it was rapidly decreased to unstimulated levels after an initial ca. 10-fold increase. The "hyperstimulation" of ODC in the absence of free arginine was reduced to normal in the presence of citrulline, the stimulation was decreased even below normal levels in the presence of ornithine and polyamines decreased ODC activity to zero levels. O delta T activity was stimulated more in the presence of arginine than in its absence. In both cases the stimulation was enhanced in the presence of polyamines and only in the absence of arginine--by ornithine.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

[Monoclonal antibodies against determinants on human MHC class II antigens].

Monoclonal antibodies against different non-polymorphic determinants of HLA class II antigens are described. The epitopes are present on HLA-DR and HLA-DQ gene products as detected by monoclonal antibody BL-Ia/l, and at HLA-DR and HLA-DP gene products which are recognized by monoclonal antibody BL-Ia/4. Another epitope recognized by the monoclonal antibody BL-Ia/DQ is only found on DQ molecules. The monoclonal antibodies BL-Ia/1, BL-Ia/4, and BL-Ia/5 are directed against the beta-subunit of 29 kDa/35 kDa class II heterodimer molecules. The recognized epitopes are shown to be not dependent on the association of the alpha- and beta-chains.

Antibodies, Monoclonal↗

[Characterization of murine monoclonal antibodies against the common acute leukemia antigen (CALLA)].

This study presents two murine monoclonal antibodies which react with the Common Acute Lymphoblastic Leukemia Antigen (CALLA). Both antibodies can be used for the diagnosis of common ALL (cALL). Indirect immunofluorescence studies (FACS-analysis) showed that the antibodies react with granulocytes and different human cell lines (Nalm-6, Reh, Raji, CCRF-CEM). The monoclonal antibodies BL-CALLA/1 and BL-CALLA/2 identify a single polypeptide chain of 95 kD. Both antibodies recognize the same or closely related epitope of the CALLA-molecule and are able to modulate in vitro the antigen on the CALLA-positive cell line Reh.

Animals↗

Polyamine effects on DNA-directed RNA polymerases in the ciliate Tetrahymena thermophila. In vivo- and in vitro-experiments suggesting highly specific regulative interactions.

Stimulation of growth in resting cultures of the ciliated protozoan Tetrahymena thermophila stimulated putrescine formation in a manner coordinated to transcription and partly to DNA-polymerisation. The stimulation of polyamine biosynthesis involved increased formation of the precursor L-ornithine from L-arginine as well as stimulation of the regulative key enzyme ornithine decarboxylase. In order to characterize the interrelationships between polyamines and transcription, which were suggested by these in vivo-assays, in vitro-assays were performed employing isolated pure macronuclei from Tetrahymena thermophila. The results obtained were: i) The diamine putrescine and the polyamines spermidine and spermine stimulated the incorporation of [4-14C]UTP into RNA time- and concentration dependently. This stimulation was not due to changes in the ionic strength nor to substitution for divalent cations (e.g. Mg2+ or Mn2+). The di- and polyamines did not alter the Km of RNA polymerases for the substrates, e.g. UTP. ii) Purified yeast-RNA, when added to the in vitro transcription system at concentrations capable of stimulating purified ornithine decarboxylase from Tetrahymena inhibited the RNA polymerases. The inhibitory effect of RNA on the polymerases was partly antagonized by spermidine and spermine but not by putrescine whereas the residual polymerase activity was stimulated by all three bases. iii) The stimulating effects of the di- and polyamines were synergistic but not absolutely additive, suggesting different targets for their actions. iv) Stimulation of RNA polymerases by putrescine, spermidine, or spermine after inhibition of particular enzymes by alpha-amanitin allowed to distinguish the effects on the three polymerases (I, II, and III): putrescine was not specific for any of the polymerases; spermidine was most active stimulating polymerase I and spermine was most active stimulating polymerase II, less active stimulating polymerase I but strongly inhibitory to polymerase III. v) The results obtained in the previous experiments were confirmed by electrophoretic analysis of the products formed in the in vitro transcription assays which furthermore showed that the differences between the experiments with or without polyamines were most pronounced if partly denatured calf thymus DNA was present in the assay mixture. This finding and the inhibition by RNA suggest that the factor influenced most by the polyamines is the binding of the RNA polymerases to the DNA target.

Amanitins↗