Seppo Partanen

Sequential appearance of relaxin, prolactin and IGFBP-1 during growth and differentiation of the human endometrium.

Relaxin (RLX) is a product of the human corpus luteum, pregnancy decidua and placenta, prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP-1) are products of the cyclic endometrium and of the pregnancy decidua. All three proteins are thought to function interdependently in endometrium/decidua as local factors within the uterus without reaching the systemic circulation. In this study, the avidin-biotin immunoperoxidase method for immunolocalization with monoclonal or polyclonal antibodies has been applied to serial sections of endometria obtained from patients at different stages of the menstrual cycle and in early and late gestation. This allowed the cellular localization of the three proteins to be followed simultaneously through the reproductive stages from cyclic endometrium to term gestational decidua. The production, as opposed to sequestration of RLX from an ovarian source was demonstrated by the application in parallel of an antibody to the processed hormone and its connecting peptide. RLX was shown localized to the glandular and luminal epithelia in the proliferative and secretory phases. The decidualized stromal cells also immunostained for RLX in the late secretory phase and in early and late pregnancy. PRL was localized first to the glandular epithelium and then stroma, appearing after RLX, IGFBP-1 appeared later in the secretory phase and predominantly in the decidualized stromal cells confirming previous studies. In contrast, all three proteins were immunostained in early pregnancy and increased to term gestation.(ABSTRACT TRUNCATED AT 250 WORDS)

Argyrophilic cells in carcinoma of the female breast

Argyrophilic hormone storage granules were sought in the tissue specimens obtained from 52 benign breast lesions (13 normal breasts, 27 cases of fibrocystic disease, 9 fibroadenomas, 2 intraductal papillomas, and 3 cases of gynecomastia) and from 90 adenocarcinomas of the female breast. No argyrophilic cells were found in the normal breast tissue or in the benign lesions studied. In three of the carcinomas (3.3%) such granules were found in the tumor cells. Using electron microscopy, the argyrophilic granules were shown to be of moderate or high electron density with an average diameter of 165 to 170 nm. Ectopic hormone production was not observed clinically in any of these three patients. The absence of argyrophilic cells in normal and benign ductal and acinar epithelium, and their occasional presence in breast carcinomas favors the concept of the histogenesis of these cells through genomic derepression during the course of neoplastic transformation.

Demonstration of catecholamines in peripheral adrenergic nerves in stretch preparations with fluorescence induced by aqueous solution of glyxylic acid

SummaryFluorescence induced by aqueous solution of glyoxylic acid and formaldehyde-induced fluorescence of catecholamines were compared for the demonstration of peripheral adrenergic nerves in stretch preparations. Glyoxylic acid was better than formaldehyde for the demonstration of the adrenergic nerves. On the other hand, the formaldehyde was better than glyoxylic acid for the demonstration of biogenic amines in cell bodies.

Fluorescence histochemical and electron-microscopical observations on the innervation of the atrial myocardium of the adult human heart

The existence of both adrenergic and cholinergic innervation of the atrial myocardium of the adult human heart was demonstrated by means of fluorescence induced by formaldehyde or glyoxylic acid and by electron microscopy. The adrenergic fluorescing axons (1) followed the course of blood vessels as typical perivascular nerve plexuses, and (2) formed a three-dimensional fairly dense nerve net obviously not related to the blood vessels. The varicosities frequently came into close apposition on myocardial cells. Several types of nerve terminals were differentiated at electron microscopy: (1) an “adrenergic” type containing small (diameter 450–700 Å) dense-cored vesicles and usually (in various proportions) small “empty” and/or large (900–1500 Å) dense-cored vesicles, (2) a “cholinergic” type containing small (ca. 500 Å) “empty” vesicles and occasionally also some large (mean diameter ca. 1200 Å) dense-cored vesicles, (3) a “pale” type containing only a few or no vesicles, (4) a “disintegrated” type containing degenerated mitochondria, autophagic vacuoles, and occasional normal-looking mitochondria, (5) nerve terminals containing a large number of mitochondria in addition to varying vesicle populations, and (6) a (possibly baroreceptive type of) nerve terminal containing myelinlike lamellated structures. The “disintegrated” and the “pale” types of nerve terminals possibly represent different stages of axonal degeneration, or may correspond to diminution in the transmitter substance concentration under certain pathophysiologic conditions, respectively. Nerve terminals crowded with mitochondria may be sensory and involved in mechano-or chemoreceptive functions. In preliminary experiments convincing evidence was obtained that the glyoxylic acid-induced fluorescence histochemical method will be suitable for comparative studies on (human) clinical specimens, e.g., for analyzing the degree of the functional activity of the intrinsic adrenergic innervation of the myocardium under various pathophysiologic conditions. The modification which appeared most appropriate for such studies is described in detail, and is proposed for use as a standard method in other similar or related studies on human clinical series. The essential criteria for analyzing the specimens at fluorescence microscopy are suggested as well.

Mediastinal Tumors: Ultrastructural and Immunohistochemical Evaluation of Intermediate Filaments as Diagnostic Aids

The histogenesis of six mediastinal tumors was investigated ultrastructurally and immunohistochemically using monospecific antibodies against intermediate filament proteins. Four of the tumors, showing different appearances by light microscopy, displayed desmosomes and cytoplasmic tonofilaments, by electron microscopy, compatible with an epithelial thymoma. These cases also showed keratin positivity by immunofluorescence microscopy. One spindle cell tumor showed zonula adherens-type junctions, prominent collections of intermediate filaments, and abundant cytoplasmic neurosecretory granules consistent with a neuroendocrine tumor. In this tumor, neurofilaments could be demonstrated by immunofluorescence microscopy, a feature also consistent with a neuroendocrine tumor. One malignant tumor, lacking tonofilaments and desmosomes but showing a few primitive junctions, did not contain keratin but showed vimentin positivity. This suggests a mesenchymal origin and a diagnosis of primitive sarcoma. These cases illustrate the diagnostic usefulness of electron microscopy and immunohistochemical evaluation of intermediate filaments.

Studies of auricular catecholamines by fluorescence histochemistry in various heart diseases of man

A comparative histochemical and clinical study concerning the state of the intrinsic adrenergic innervation of the human atrial myocardium was carried out, using the glyoxylic acid-induced fluorescence histochemical method. Specimens from the right auricular appendage were obtained during open-heart surgery from patients suffering from 1. ischaemic heart disease (IHD), 2. atrial septal defect of the secundum type (ASD), and 3. left-sided univalvular or multivalvular heart disease (VHD) with or without congestive heart failure (CHF) experienced prior to surgery. In the IHD group the densities of both the perivascular and the “free” myocardial adrenergic nerve net were greater than in the ASD group and especially in the VHD/ CHF group. Secondly, the intensity of fluorescence of the adrenergic structures was generally higher in the IHD group than that in the VHD/CHF group. Further, the average size of the varicosities, the number of varicosities per given length of axon, and the proportional share of the large varicosities were greater in the IHD group than in the ASD and VHD/CHF groups. The difference between the IHD and ASD groups was not great but was obvious in any case. In some patients with VHD/CHF fluorescing axons were observed only occasionally, and the tiny varicosities exhibited a hardly discernible fluorescence. Thus the amount of noradrenaline (NA) in the adrenergic fibres in the IHD group seems to be higher than in the ASD and especially VHD/CHF groups. The high level of NA in the IHD group is assumed to constitute a contributory factor in both intracellular metabolic changes and the systemic changes typical of myocardial ischaemia and infarction. In one patient with IHD and in six patients with VHD/CHF with significantly higher heart volume (mean±SD) compared with the rest of the patients (P<0.001), huge local axonal accumulations of NA in the form of “droplet fibres” were found. These enlarged, bulging adrenergic axons are assumed to be a consequence of mechanical trauma with stretching or disruption of the axons due to myodegenerative processes. It is further assumed that these “droplet fibres” are relatively common in those patients with diseased myocardium. They may constitute an extra contributory factor to the tendency to arrhythmias so typical of patients of this kind, by increasing the excitability of non-automatic tissue.

Evolution of substance P immunofluorescent nerves in callus tissue during fracture healing

Tibial fractures were experimentally produced in rats. The fractures were allowed to heal spontaneously. Samples of callus tissue were studied by a specific immunohistochemical method for substance P, the peptide which is found in sensory nerves and probably plays a role in nociceptive nerve transmission. The control periosteum contained a rich network of substance P immunofluorescent nerves. The early callus tissue also contained such sensory nerves, the peak frequency of which was at the end of the second week. It is proposed that the substance P-containing nerves play a protective nociceptive role during fracture healing.

Effects of steroid hormones on the differentiation of the catecholamine storing cells of the paracervical ganglion of the rat uterus

SummaryThe effect of hydrocortisone, prednisolone, estrogen, progesterone and testosteron on the differentiation of the paracervical ganglion of the rat uterus was examined by the formaldehyde induced fluorescence (FIF) method. Prednisolone and hydrocortisone treatment caused about a tenfold increase in the number of small, intensively fluorescent (SIF) cells. The SIF-cells were larger and organized in larger groups than in untreated animals. The SIF-cells were also arranged in a satellite cell position around the ganglion cells, which were non-fluorescent or showed a weak to moderate FIF. The amount of solitary SIF-cells was increased. No effects detectable by the FIF-method were found with certainty after administration of estrogen, progesterone or testosterone.The effect of glucocorticoids on the differentiation of autonomic ganglia is discussed. It seems probable that the main part of the developing SIF-cells after glucocorticoid treatment develop from undifferentiated neural cells.

Histochemically demonstrable increase in the catecholamine content of the carotid body in adult rats treated with methylprednisolone or hydrocortisone.

SynopsisCarotid bodies of adult albino rats were examined using the formaldehyde-induced fluorescence (FIF) method for the demonstration of fluorogenic monaomines and staining with I% Toluidine Blue for morphological observations.In the carotid body of normal controls, most glomus (principal or type I) cells exhibited a FIF presumably due to catecholamines. The intensity of the fluorescence was weak in most cells, while some glomus cells were non-fluorescent and others exhibited a moderate or intense FIF. The sustentacular (satellite, supporting or type II) cells were essentially non-fluorescent.One week after the administration of a single intraperitoneal injection of the long-acting glucocorticoid 6-methylprednisolone sypionate (400 mg/kg) or after seven intraperitoneal injections of the water-soluble glucocorticoid hydrocortisone sodium succinate (40 mg/kg daily for a week), a distinct increase was observed in the FIF of the glomus cells. No non-fluorescent glomus cells were observed after treatment with either glucocorticoid, and the intensity of most fluorescent glomus cells was moderate or intense.It is concluded that glucocorticoids cause an increased storage of catecholamines in the glomus cells of the carotid body of the adult rat, an observation of interest in view of the fact that such changes due to glucocorticoids have as yet been reported only in catecholamine-storing cells of newborn rats.

Monoamine—Containing structures in the hypothalamo-hypophyseal system in the human fetus

SummaryThe monoamine containing structures of the human fetal hypothalamohypophyseal tract were studied histochemically by demonstrating the formaldehyde induced fluorescence. First specific fluorescence in the hypothalamus and median eminence was observed at the age of 15 weeks. A sparce network of fluorescent nerve fibres was found to line the walls of the ventricle. Weakly fluorescent cells were found within the stromal cell cords of the adenohypophysis at the age of 11 weeks. Both the number and the fluorescence intensity of these cells increased up to the age of 15 weeks. The apperance of the monoamines in the cytoplasm of the adenohypophyseal cells correlates well with the previous observations on the onset of the function of the fetal adenohypophysis. The lack of monoaminergic innervation of the hypothalamus and median eminence suggests that the hypothalamoadeno-hypophyseal neurohumoral regulation might not be functional during the first trimester of pregnancy.