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Biomedical subjects

D Grube

Publications and source records attributed to D Grube.

At least 73 records · Page 4Linked to original sources

The Merkel cell--a member of the APUD cell system. Fluorescence and electron microscopic contribution to the neurotransmitter function of the Merkel cell granules.

Merkel cells of sinus hair follicles of nude mice were investigated by fluorescence and electron microscopy following pretreatment with amine precursors (L-Dopa, L-5-Hydroxytryptophan) and monoaminoxidase-inhibitors (Marsilid, Harmaline). Neither in control animals nor in pretreated animals any evidence for an involvement of Merkel cells in monoamine metabolism could be found. Therefore, the hypothesis that Merkel cells might share the most constant cytochemical characteristics of the cells of the APUD series, i.e., to take up amine precursors, should definitely be left. Following Harmaline treatment, however, Merkel cells were found fluorescent; cytophotometric analysis of the fluorescence emission spectra of formaldehyde-treated tissue ascertained that this fluorescence was due merely to a specific Harmaline fluorescence. The significance of Harmaline uptake in Merkel cells, most probably in the Merkel cell granules, is discussed.

5-Hydroxytryptophan↗

Pancreatic glucagon cells contain endorphin-like immunoreactivity.

Semithin and thin serial sections of rat pancreas were investigated by immunocytochemical techniques using antisera against glucagon and endorphin. The immunocytochemical staining methods revealed that in the rat endocrine pancreas endorphin-like immunoreactivity is contained in glucagon (A-) cells. All secretion granules of glucagon cells that could be identified in adjacent thin sections exhibited glucagon as well as endorphin immunoreactivity. Since glucagon cells are devoid of ACTH-like immunoreactivity it is concluded that the biosynthetic pathway of pancreatic endorphin is not the same as in the pituitary.

Animals↗

[Radioimmunoassay [RIA] of cholecystokinin-pancreozymin [33-CCK] (author's transl)].

A further specific, reproducible radioimmunoassay of cholecystokinin-pancreozymin was developed. 125J-labelled 39-CCK was used as tracer. The limit of detection for CCK in serum was 40 pg/ml. Basal levels for CCK have been found to be 222 pg/ml, being increased after food ingestin. No crossreactivity with gastronintestinal hormones could be observed. Somatostatin was inhibiting the stimulated CCK-release. Histochemical and radioimmunological determinations localized CCK-containing cells in duodenal tissue of humans. Physiological and pathological processes in which CCK is involved have to be studied to evaluate its role in the gastrointestinal tract.

Cholecystokinin↗

Estimation of cholecystokinin-pancreozymin (CCK) in human plasma and tissue by a specific radioimmunoassay and the immunohistochemical identification of pancreozymin-producing cells in the duodenum of humans.

A reliable, sensitive, reproducible and specific radioimmunoassay for cholecystokinin-pancreozymin (CCK) has been developed, using rabbit antisera to highly purified porcine hormone. The natural occurring variant of CCK (39-CCK), in which the ordinary CCK is lengthened from its N-terminus by a hexapeptide, labelled with 125J, and repurified by column chromatography on Sephadex G-10 and on SP-Sephadex C-25, was used as tracer. Separation from antibody-bound labelled 39-CCK was carried out using a double antibody procedure. Non-specific interference with the assay system was abolished by ethanol extractions. Highly purified porcine CCK was used as standard. No significant crossreaction was found with gastrin, motilin, vasoactive polypeptide (VIP), gastric inhibitory polypeptide (GIP), natural and synthetic secretin, pancreatic glucagon or insulin. The sensitivity of the assay is approximately 40 pg/ml of test solution. The mean immunoreactive CCK concentration in 45 fasting normal subjects was 222 pg/ml increasing after food ingestion to 480 pg/ml. Somatostatin was able to abolish the stimulated CCK release. Elevated CCK concentrations were found in chronic pancreatitis. Immunohistochemical identification of pancreozymin cells was carried out either in surgical samples or in biopsy material. Approximately 1650 CCK cells per cross-section in the duodenum of humans have been found. The CCK cells usually appeared elongated, oval or pyramidal in shape and were observed to reach the lumen with their apical cell pole.

Animals↗

A carcinoid occurring in the testis.

Carcinoids of the testis are rare tumours developing in three different ways: 1. It may differentiate within a teratoma, 2. it may be a metastasis of a "loco alieno" seated carcubiud abd 3. it may represent a real primary carcinoid. The observation of a primary testicular carcinoid in a man aged 55 years afforded the opportunity to study such a tumor for the first time by electron microscopic and fluorescence microscopic methods. Thereby, it could bw shown, that this testicular carcinoid corresponds to the carcinoids of the lower small gut. According to the specific ultrastructure of the intracytoplasmic granules it must derive histogenetically from an EC-cell. At the moment it cannot be decided whether the primary testicular carcinoid represents an autochthonous tumor of the male gonad or solely a teratoma with one-sided differentiation in the sense of a simplified teratoma.

Carcinoid Tumor↗

[The endocrine cells of the gastrointestinal epithelium and the metabolism of biogenic amines in the gastrointestinal tract (author's transl)].

After a review on the historical development of morphological investigations of entero-endocrine cells, dating back to 1870, a detailed synoptical review of the current stage of findings in this field is given. At the present time nine different endocrine cell types can be distinguished in the epithelium of the gastrointestinal tract. Criteria for this differentiation are properties concerning specific staining methods, aldehyde-induced fluorescence, immunohistochemistry, and ultrastructure. From present results it is obvious that distinct cell types are responsible for the synthesis of defined polypeptide hormones (e.g. gastrin, secretin, enterogastrone). The metabolism of amines, in relation to the endocrine cells of the gastrointestinal tract is of particular interest here. Points investigated include the uniqueness of endocrine cells, with regard to the metabolism of biogenic amines ("APUD-cells") and the possibility of serotonin synthesis by a definite cell type, i.e. by the EC-cell ("enterochromaffin" cell). In our experimental animal, male Wistarrats, seven different entero-endocrine cell types can be discerned by ultrastructural means: EC-, ECL-, G-, AL-, EG-, D- and D1-cells. The I-cell (found in other species) can hardly be distinguished from the AL-cell by ultrastructural means and the S-cells, as found in other species, are not to be found at all. Only some of the cited cell types can be seen by fluorescence microscopy. After formaldehyde-treatment of the tissue, the "enterochromaffin" cell shows a yellow, serotonin-specific fluorescence. This cell corresponds in shape, number and distribution to the ultrastructurally defined EC-cell. EC-cells are found predominantly in the pyloric region and the duodenum and less frequently in the middle- and hindgut and the cardiac region; seldomly EC-cells are encountered in the oxyntic gland area of the stomach. In the rat gastro-intestinal tract, number and fluorescent intensity of EC-cells does not always correspond with the serotonin content of a certain region--sometimes the level of serotonin is largely determined by the mast cells, which in the rat also contain serotonin. For example, the high serotonin content of the oxyntic gland area, which contains very few EC-cells, has to be contributed nearly exclusively to mast cell serotonin. Mast cells can be domonstrated by fluorescence microscopy, due to their histamine content, after treatment of the tissue with o-phthalaldehyde (OPD). It seems likely that the histamine content, especially that of the so-called "atypical mast cells" of the mucosa, is inversely related to their respective serotonin content. --In addition to mast cells, OPD-treatment leads to a fluorescence in some of the entero-endocrine cells of the gastrointestinal epithelium. In the gastric epithelium these fluorescing cells should be regarded as histamine-containing ECL-cells and glucagon-containing AL-cells while in the colonic epithelium they are considered to be glucagon-containing AL-cells...

5-Hydroxytryptophan↗

Differential demonstration of the motilin-cell and the enterochromaffin-cell.

Motilin cells were investigated in the GI-tract of various laboratory animals by immunfluorescence. The distribution of motilin cells was compared to enterochromaffin (EC) cell localization. Simultaneously histochemistry for EC-cells and motilin cells was carried out. The results indicate that motilin is not produced within the EC-cells. Present classification of motilin cells on ultrastructural evidence is therefore not yet possible.

Animals↗

Effects of guanethidine on paraganglionic cells in the superior cervical ganglion of the rat.

Paraganglionic cells in the rat superior cervical ganglion were investigated by fluorescence and electron microscopy following treatment with guanethidine for 5-30 days. Control animals received saline and guanidine. Fluorescence cytophotometric measurements revealed a general decrease in the catecholamine content of paraganglionic cells in guanethidine-treated animals. However a few cell clusters showed focal increases. Similarly by electron microscopy there was a general decrease of cell clusters showing increases. -- guanethidine -- as well as guanidine--treated animals showed non-specific cytological alterations such as mitochondrial swelling and increase of cytoplasmic glycogen. However no changes of catecholamine contents and of dense core vesicles were noted in control animals. These results confirm the conception that in rat paraganglionic cells the dense core vesicles are the main storage site of catecholamines. The marked difference in the response of some cell culsters to the experimental treatment can be considered as evidence of functional heterogeneity of this cell population in the rat superior cervical ganglion.

Animals↗