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G Vollmer

Publications and source records attributed to G Vollmer.

At least 73 records · Page 4Linked to original sources

Tenascin expression in the human endometrium and in endometrial adenocarcinomas.

To investigate the involvement of tenascin, an extracellular matrix glycoprotein, in epithelial growth and malignancy, its specific distribution pattern in the human uterus was examined immunohistochemically. During the proliferative phase of the menstrual cycle, this antigen was found as a sharp band around the endometrial glands. The immunoreactivity persisted until the early postovulatory phase of the menstrual cycle, but was not detectable in the glandular or stromal compartment during this later secretory stage, instead endometrial arterioles were immunostained. In marked contradistinction, when antibodies directed against tenascin were applied to sections of endometrial adenocarcinoma, almost the entire extracellular space stained, whereas the neoplastic cells themselves were nonreactive, whatever the degree of tumor differentiation. In precancerous proliferative lesions of the endometrium, tenascin's presence was variable. It was detectable around some superficial glands demonstrating cystic hyperplasia and around all deeply situated glands at the endometrial/myometrial interface. In cases of adenomatous hyperplasia, tenascin immunolocalized throughout the extracellular space of the stroma and the staining intensity was increased as the hyperplasia became more atypical. We therefore conclude that tenascin may be a stromal marker for epithelial proliferative states including those associated with malignancies of the endometrium.

Adenocarcinoma↗

Relationship of cytosolic estrogen and progesterone receptor content and the growth fraction in human mammary carcinomas.

In this study of breast cancer specimens, the relationship between cytosolic estrogen (ER) and progesterone receptor (PR) content to the size of the respective growth fraction (GF) (expressed as percentage of proliferating tumor cells) was investigated. We applied the recently developed ligand-binding assay for extracts of frozen sections and Ki-67 immunocytochemistry for the assessment of the GF to adjoining serial sections of a single tissue block. If the receptor content is plotted against the percentage of Ki-67 labeled cells, an inverse relationship between receptor content and proliferation becomes obvious, meaning that a high receptor content is associated with a small GF and vice versa. If tumor specimens are grouped according to the evaluated receptor status, the mean percentage of Ki-67-positive cells is 12% for ER-positive/PR-positive (ER+/PR+), 26% for ER-positive/PR-negative (ER+/PR-), 55% for ER-negative/PR-positive (ER-/PR+), and 57% for ER-negative/PR-negative (ER-/PR-) specimens. A significant population of tumors exists, however, which exhibit a high receptor content and a high GF. The percentages of ER+/PR+ samples with a high proliferation index are 16 and 26% if the total ER+ population is considered.

Biomarkers, Tumor↗

Photoaffinity labelling of gonadotropin releasing hormone binding sites in human epithelial ovarian carcinomata.

A photoaffinity labelled derivative of [D-Lys6]-GnRH was prepared with a bifunctional photolabile reagent (4-azidobenzoyl)-N-hydroxysuccinimide. In rat pituitary membranes, this analog retained high binding affinity (Ka = 0.12 x 10(9) M-1) consistent with a single class of receptors. The analog was iodinated and used for the identification of GnRH binding sites in human epithelial ovarian carcinomata. By sodium dodecyl sulfate electrophoresis in 10% polyacrylamide gel the presence of two labelled components could be demonstrated: a high molecular weight component of 63,200 and a smaller component of 46,000. Competition experiments with unlabelled ligand suggest that it is the high molecular weight component which specifically binds GnRH.

Adenocarcinoma↗

[Should one teach false theories: a reflection on the teaching of science].

Consciously teaching wrong theories seems to go against both the ethics of science and the duties of the responsible educator. Even so, it can be shown that we regularly teach wrong things, e.g., Galileo's laws or classical mechanics in general. And that we are even justified in doing so if some specified conditions are met. One of these five conditions is that the theory taught is, though false, still useful enough in other respects.

Ethics, Professional↗

Quantification of estrogen- and progesterone receptors from cryostat sectioned human tumor tissue: comparison of ligand binding and immunocytochemical assays.

We have recently described an assay procedure to measure estrogen and progesterone receptors in extracts from frozen sections by a ligand binding assay. With this methodological approach it is now possible to perform comparative experiments not only to DCC/Scatchard analyses from different tissue blocks, but also to immunocytochemical determinations in identical tissue blocks: (1) When receptor quantities measured by the two biochemical methods were compared, a high correlation of estrogen receptor content was found between determinations in supernatants from frozen sections and DCC/Scatchard analyses. A slightly poorer correlation in the comparison of the ligand binding assays was obtained for the progesterone receptor. (2) The percentage of tumor cells stainable by immunocytochemistry for estrogen and progesterone receptors could hardly be correlated to the receptor concentrations (fmol/mg) measured quantitatively by the two ligand binding assays. (3) As the final result tumor specimen could be grouped into classes of receptor status according to the presence or absence of a nuclear stain in immunocytochemical assays or according to receptor concentrations above or below distinct threshold which were fixed at 20 fmol/mg for Scatchard analyses of both receptor species, 20 fmol/mg for estrogen- and 40 fmol/mg for progestin receptors for the assay with sections. In this diagnostical consideration the concordance of both biochemical methods to the immunocytochemical assessment was high for estrogen and less pronounced for progesterone receptors. (4) In some breast cancer specimen analyzed biochemically an unspecific progestin binding component could be detected superimposed on the progesterone receptor peak after isoelectric focusing.

Breast Neoplasms↗

Simultaneous quantification of oestrogen and progesterone receptors by a ligand binding assay in frozen sections.

We describe two modifications of a double-isotope assay for measuring the concentrations of oestrogen receptors and progesterone receptors in tumour cytosols and extracts of frozen tumour sections and endometrial sections. The concentrations of these receptors are derived from single-point/isoelectric focussing assays after incubation of cytosols or section supernatants either with ([125I]vinyl)-nortestosterone and [3H]oestradiol or [125I]oestradiol and [3H]ORG 2058 (16 alpha-ethyl-21-hydroxy-19-nor[6,7(3)H]pregn-4-ene-3,20-dione). The concentrations of the oestrogen and progesterone receptors found in cytosols (r greater than 0.93) and extracts from sections (r greater than 0.8) by dual label assays are highly correlated with those found by single label assays. The method described represents an approach to the determination of oestrogen and progesterone receptors biochemically in an amount of tissue which is comparable to that needed for immunocytochemical procedures.

Breast Neoplasms↗

An improved micromethod for the determination of biochemically active estrogen and progesterone receptors in parallel to comparative histological examination of a single piece of tissue.

An improved radioreceptor assay of unfixed cryostat sections of human target tissues has been developed. Sections collected on glass coverslips were immediately incubated with 5 nM concentrations of either tritiated estradiol-17 beta for estrogen receptor (ER) or ORG 2058 for progesterone receptor (PR) determination. For quantitation, receptor-bound and free hormone were separated by isoelectric focusing (IEF). The assay allows the determination of steroid hormone receptors and comparative histological examinations in immediately neighbouring serial sections of a single piece of tissue. Biochemically, the validity of the assay procedure was evidenced by Scatchard analysis, by ligand and tissue specificities, by the linear relations of receptor and protein concentrations and the number of sections per test tube. Diagnostically, we compared the routine (6 point DCC-Scatchard) procedure for breast cancer analysis with the section method. A good correlation for ER and a less pronounced correlation for PR was found. Statistically, the precision of the method was verified by low deviations of duplicate determinations, low day-to-day variations and low inter-assay variations.

Breast Neoplasms↗

Quantitative determination of oestradiol-17 beta hydroxysteroid dehydrogenase: increased sensitivity by HPLC separation of the hormones permits the measurement of enzyme activity in cryostat sections.

The activity of the oestradiol-17 beta hydroxysteroid dehydrogenase in human endometrial and breast cancer specimens was determined by the NAD-dependent conversion of oestradiol-17 beta to oestrone. The sensitivity of the determination was improved by the separation of the hormones by HPLC. We are now able to determine oestradiol-17 beta hydroxysteroid dehydrogenase quantitatively in cryostat sections. A clear correlation of serum progesterone levels and oestradiol-17 beta hydroxysteroid dehydrogenase activity in the endometrium was demonstrated, and we found a more than 30-fold increase in enzyme activity after the progesterone surge. In contrast, in breast cancer samples, we found no correlation between oestradiol-17 beta hydroxysteroid dehydrogenase and the measured serum parameters.

17-Hydroxysteroid Dehydrogenases↗

Cholinesterases and cell proliferation in "nonstratified" and "stratified" cell aggregates from chicken retina and tectum.

Dissociated single cells from chicken retina or tectum kept in rotation-mediated cell culture aggregate, proliferate and establish a certain degree of histotypical cell-to-cell relationships ("sorting out"), but these systems never form highly laminated aggregates ("nonstratified" R- and T-aggregates). In contrast, a mixture of retinal plus pigment epithelial cells forms highly "stratified" aggregates ("RPE-aggregates", see Vollmer et al. 1984). The present comparative study of "stratified" and "nonstratified" aggregates enables us to investigate the process of cell proliferation uncoupled from that of tissue stratification. Here we try to relate these two basic neurogenetic processes with patterns of expression of cholinesterases (AChE, BChE) during formation of both types of aggregates. During early aggregate formation, in both "stratified" and "nonstratified" aggregates an increased butyrylcholinesterase activity is observed close to mitotically active cells. Quantitatively both phenomena show their maxima after 2-3 days in culture. In contrast, AChE-expression in all systems increases with incubation time. In nonproliferative areas, in the center of RPE-aggregates, the formation of plexiform layers is characterized initially by weak BChE- and then strong AChE-activity. These areas correspond with the inner (IPL) and outer (OPL) plexiform layers of the retina in vivo. Although by sucrose gradient centrifugation we find that the 6S- and the fiber-associated 11S-molecules of AChE are present in all types of aggregates, during the culture period the ratio of 11S/6S-forms increases only in RPE-aggregates, which again indicates the advanced degree of differentiation within these aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholinesterase↗

Reaggregation of chick retinal and mixtures of retinal and pigment epithelial cells: the degree of laminar organization is dependent on age.

We report here a striking age dependency for tissue reconstruction capacity in rotating cultures of chick retinal cells, in particular in mixtures of retinal and pigment epithelial cells. The comparison of sections after acetylcholinesterase histochemistry delivered the following results. In aggregates from retinal cells, sorting out is clearly observable, but only a minor age-dependent effect is detectable, e.g. the laminar order of aggregates derived from 6-day-old cells is increased compared to aggregates from other stages. In aggregates from mixtures of retinal and pigment epithelial cells, the degree of laminar order is highest with cells from 5-day-old embryos and decreases significantly at older ages. The highest number of aggregates with the reconstruction of all main retinal layers was obtained by the reaggregation of retinal cells of embryonic day 5 (25%). To achieve laminar order in aggregates, at all ages, the addition of pigment epithelial cells seems to be a prerequisite.

Acetylcholinesterase↗

Lucifer Yellow stains a subpopulation of chicken retinoblasts preferentially 3 h after their last round of DNA synthesis.

Observations of histological preparations of E4-E6 retinae suggested to us that Lucifer Yellow (LY) uptake occurs during a specific phase of the mitotic cell cycle. We examined this phenomenon more closely in intact E6 retinae. If we first exposed the tissues to a 20-min pulse of tritiated thymidine followed by LY-staining at different intervals, a distinct peak of double-labelled cells was found 3 h after exposure of the tissue to thymidine. In contrast, if staining with LY is followed by thymidine pulses, only few double-labelled cells are found. We conclude that a subpopulation of young neurons takes up LY specifically after their last round of DNA-synthesis.

Animals↗

An in vitro model of proliferation and differentiation of the chick retina: coaggregates of retinal and pigment epithelial cells.

Pigment epithelial (PE) cells exert a pronounced organizing effect when added to embryonic day (E) 5-6 chick retinal cells in a reaggregation system such that after a period of 14-21 d of culture, the main layers of an intact E10-E14 retina are reconstructed (Vollmer et al., 1984). In the present study we investigated the time course of the formation of retina-like structures in retinal-pigment epithelial aggregates, in particular, the fate of the PE cells and their influence on processes of differentiation within the aggregates. PE cells first form a core in the center of the aggregates and migrate to the periphery at later stages. The PE core affects the organization of proliferation and differentiation, phenomena that were monitored using 3H-thymidine autoradiography and AChE histochemistry, respectively. A double-staining procedure combining both techniques on the same section is described. Soon after aggregation, a matrix zone and a zone of differentiated cells are formed. At later steps, proliferation becomes gradually restricted to a narrow band within the aggregates comparable to the in vivo situation. The spatiotemporal pattern of withdrawal from mitosis resembles that of the in vivo retina. Proliferation in aggregates is sustained over a longer period with PE, as compared with aggregates formed by retinal cells alone. AChE staining in aggregates in the presence of PE shows a layered appearance, while there is only crude sorting-out of labeled and unlabeled cells in aggregates composed of retinal cells only. The basis of PE cell action as well as the relevance of this in vitro system for understanding normal eye development are briefly discussed.

Animals↗

Reaggregation of embryonic chick retina cells: pigment epithelial cells induce a high order of stratification.

We report here that, in comparison to aggregates from retinal cells alone, addition of pigmented epithelial cells to retinal cells in rotary culture results in a pronounced increase of spatial order. A particularly high level of organization is found in about 15-20% of the aggregates. In these 'retinoids' the main layers characteristic of developing in vivo retinae can be distinguished in correct sequential arrangement on the basis of morphological criteria and by using acetylcholinesterase histochemistry [5, 6, 15], peanut agglutinin-lectin binding [11] and Lucifer Yellow staining [7-9].

Acetylcholinesterase↗

Selective uptake of Lucifer Yellow into different cell populations of the developing chicken retina.

Whole chick embryo eyes at different stages of development were incubated in a solution of Lucifer Yellow (LY) and examined in frozen sections. Different cell populations selectively take up the dye during the course of development: from embryonic day 4 - day 8 of incubation (E4-E8) only a subpopulation of neuroepithelial cells are stained, which at E7-E8 are located mainly near the periphery between the prospective ora serrata and the outer edge of the retina. This staining disappears after E8. A second phase starts at E10, with a weak but specific staining of a small population of cells in the outer nuclear layer which becomes less pronounced by E12. Starting around E10.5 a bright staining of cells in the innermost part of the inner nuclear layer can be observed. Their size, shape and location suggest that most of them represent a subclass of amacrine cells (including all 'displaced amacrine cells'). Beginning at E15, the staining pattern becomes more complex. A single cell analysis of the percentage of LY-stained cells which are stained either before or after papain-treatment and dissociation of the intact tissue confirms and extends the histological findings.

Animals↗

Lucifer yellow stains displaced amacrine cells of the chicken retina during embryonic development.

We have used Lucifer Yellow for histological tracing of displaced amacrine cells within the ganglion cell layer (GCL) during the embryonic development of the chicken retina. Incubating whole eyes in the dye leads to bright staining of all displaced amacrine cells, whereas ganglion cells and glial cells are not stained. A subpopulation of cells of the inner part of the inner nuclear layer (INL) are also stained (for further details see ref. 13). Kainic acid, which is known to interfere with and kill amacrine cell systems, blocks the staining of these cells fully. This in addition to histological evidence confirms that the LY-stained cells in the GCL are displaced amacrine cells. Of the cells in the GCL, 23% (+/- 3%) are of the displaced amacrine type. Further, we find that the cytoarchitectural arrangement of these cells changes significantly during development.

Animals↗

[On the diuretic and saluretic activity of p-hydroxytriamterene and methoxytriamterene (author's transl)].

Applying i.v. the phase-I-metabolite of 2,4,7-triamino-6-phenylpteridine (triamterene), p-hydroxytriamterene, to rats, we found a strong natriuresis. This contradicts the investigation of a previous study which stated that p-hydroxytriamterene is essentially inactive. We think, these different results depend on the fact that probably oral application was employed without testing the absorption concerned.

Animals↗