Histology and histopathology Vol. 3, nº 3 (1988)

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The influence of matrix components on the morphological differentiation of a proliferating hepatocyte line from liver of newborn mice
(Murcia : F. Hernández, 1987) Lilja, S.; Merker, H. J.; Ghaida, J.
The differentiation behaviour of a liver epithelial cell line of the newborn mouse cultured on various matrix components (Thermanox pure. Thermanox coated with ECM, dried collagen type I and type 11, wet collagen type I and type I11 and on floating collagen) was investigated by electron microscopy. Only during the last few days of pregnancy and up to day 9 p.p. could these cells be isolated using a very delicate method. The cells were smaller than differentiated hepatocytes and proliferated spontaneously. They resembled the socalled oval liver cells. On Thermanox pure or Thermanox coated with ECM, dried collagen type I or type I1 a confluent monolayer developed after about 6 days that consisted of rather flat extended cells which were characterized by short contacts and the absence of any morphological indications of differentiation. On wet collagen the extension area was smaller and the cells were taller. The length of the contact area and the number and size of gap junctions and cell organelles increased. On floating collagen multi-layered aggregates of polygonal cells developed that were characterized by extended cell contacts, bile capillary-like structures and highly developed cell organelles, especially rough endoplastnic reticulum Since differentiation processes can be demonstrated ultrastructurally only on wet collagen, especially on floating collagen, the chemical composition of the sub\trate and a specific matrix-cell interaction cannot be the only triggering factor. It is assumed that mechanical properties of the suhstrate, e.g. plasticity, are involved. The change in the shape of the cell, the prolongation or intensification of the cell contact and the adaptation of the cytoskeleton might play a decisive role in this connection.
Open Access
Proteoglycans of alveolar bone of diabetic and non-diabetic mice, a histochemical study
(Murcia : F. Hernández, 1987) Johnson, R. B.
The effect of diabetes mellitus on the interdental alveolar bone has been long debated. The present study reported the distribution of glycosaminoglycans (GAG) in normal and diabetic alveolar bone using histochemical techniques. Animals were rendered diabetic and killed at 2,4,6 and 9 weeks after injections. Tissues were stained with Alcian blue 8GX dye (pH 2.5) to demonstrate GAG and the intensity of the staining reactions compared with age-matched controls. During the experiment, weights of control animals did not change significantly; weights of diabetic animals were significantly less than initial weights from O- 6 weeks (p<0.001), but became nearly equal by 9 weeks. Staining intensity of diabetic bone demonstrated initial decrease (0-4 weeks) followed by a marked increase (4-9 weeks) suggesting an early decline in bone GAG levels followed by increased bone GAG levels as compared to age-matched control and initial levels. Bone GAG levels were significantly different between diabetics and agematched controls at 2 (p<0.005) 4 (p<0.001), 6 (p<0.001) and 9 (p<0.001) weeks after streptozotocin injections. Digestion with chondroitinase AC, ABC and streptomyces hyaluronidase suggested significant differences between control and diabetic bone matrix in the levels of chondroitin 4 and 6 sulfates (p<0.05) and hyaluronic acid (p<0.001) but not dermatan sulfate. In control and diabetic bone, chondroitin sulfates were located within the bone matrix, dermatan sulfate within bone matrix and Sharpey fiber bundles. Thus, bone GAG changes in diabetes mellitus occur predominately within bone matrix and are variable depending on the length of the disease.
Open Access
Metaplasia of the parietal layer of Bowman's capsule in the human kidney. Incidence in alcoholic liver disease and hypertension
(Murcia : F. Hernández, 1988) Haensly, William E.
This report is the second of two surveys to determine the incidence of metaplasia of Bowman's parietal epithelium in the human kidney. Human kidney sections obtained at autopsy at the Department of Pathology, University of Texas Medical Branch, Galveston, Texas, were examined with the light microscope. The kidneys were fixed in neutral formalin, sectioned at 6 pm and stained with hematoxylin and eosin. Autopsy records were consulted after kidney section examination to determine if there was any correlation between clinical disease, histopathological changes in organ systems and metaplasia of Bowman's capsule. The kidney sections represented both sexes in 8 age groups, from less than one year to 80 years. A total of 174 kidneys, representing 174 individuals, were evaluated. One hundred renal corpuscles were counted per section and the parietal layer of Bowman's capsule was classified as normal (squamous) or metaplastic (cuboidal). Of the 174 kidneys examined, 137 (79% )- 79 male and 58 female - had metaplasia of Bowman's capsule. On the average, in the kidneys with the lesion, 6% of the renal corpuscles had metaplasia of Bowman's parietal layer. The lesion was present in both sexes in all age groups. The autopsy records revealed that metaplasia of Bowman's parietal epithelium was usually present with hepatic fatty changes andlor congestion. Alcoholic liver disease and hypertension represented the most frequent clinical diseases in the sample; these conditions had the highest incidence of metaplasia. Twenty-six of the 174 kidney samples were from individuals with alcoholic liver disease, all of whom had metaplasia of Bowman's capsule. In the latter kidneys the mean percentage of glomerular capsules with metaplasia was 14%, with a range of l to 46%. Thirtyeight of the 174 kidney samples were from individuals with hypertension, 29 (76% ) of whom had metaplasia of Bowman's capsule. In these kidneys the mean percentage of glomerular capsules with metaplasia was 6% with a range of 1 to 16%. These observations suggest that metaplasia of Bowman's parietal epithelium is a common occurrence in the human kidney under different pathological conditions, and is especially prominent in alcoholic liver disease and hypertension.
Open Access
Pituitary growth hormone secretory granules in streptozotocin-induced diabetic rats
(Murcia : F. Hernández, 1988) Masataka Shiino; Toshiko Ishibashi
The ultrastructural features of growth hormone (GH) secretory granules were observed by immuno-gold procedure in the anterior pituitary glands of streptozotocin (STZ) - induced diabetic rats. The diabetic state of rats induced by one injection of STZ was severely advanced at 10 months after the injection. The cytoplasmic organella were revealed to be poorly developed in GH-cells of the diabetic rats. The size of immunoreactive GH-granules of 10-months-old diabetic rats was smaller than that of sham-controls. The insulin therapy in the diabetic rats resulted in a considerable improvement in relation to the changes seen in the GHcells, but the therapy did not return the profile of the granule size distribution to that of the control group
Open Access
Eosin-related fluorescence of acidophil pituitary cells
(Murcia : F. Hernández, 1988) Friedman, H.; Friedman, I.V.C.; Mello, C.V.
The examination of haematoxylin and eosin stained sections of normal and neoplastic pituitary glands under ultraviolet light illumination discloses fluorescence of acidophil cells. The distinction between prolactin and growth hormone-producing cells is not possible. Such fluorescence depends on previous eosin staining.
Open Access
The fibrogenic response of adult rat lung to continuous propranolol treatment
(Murcia : F. Hernández, 1988) Smith, Dennis M.; Sommers Smith, Sally K.
Fibrogenesis is a common pulmonary response to injury, which is usually preceded by other severe reactions, including inflammation, fluid exudation, and alveolar epithelial damage and proliferation. The purpose of this study was to examine the morphologic effects on the distal lung of a continuous propranolol treatment. Adult male rats were treated, via a subcutaneous osmotic pump, with a continuous (approximately 0.5 mglhour) dose of propranolol HCl, a potent wide range beta-adrenergic blocking agent, in saline, or saline alone. The animals were killed after one week or three weeks. Electron microscopy of the lungs of the propranolol-treated animals revealed a dramatic increase in the prominence of interstitial cells and fibers of the alveolar septa, along with focal thickening of endothelial cells and some morphologic changes in type I1 alveolar epithelial cells. In some animals an analysis of total protein content, as well as 3H-proline incorporation into total protein and collagen was undertaken. The results of this study indicated a significant increase in total protein content and proline incorporation into collagen in the lungs of animals treated for seven days with continuous propranolol. There was no evidence of stimulated blood cells, macrophages, edema or severe epithelial damage. This study provides morphologic evidence that continuous treatment with moderate levels of propranolol results in a fibrogenic response in the peripheral lung, in the absence of typical hallmarks of severe pulmonary damage.
Open Access
Maternal smoking during pregnancy affects neuroendocrine cells in the neonate hamster lung
(Murcia : F. Hernández, 1988) Keith, lngegerd; Kary, John R.
Primigravid Syrian golden hamsters were exposed in a Walton smoking machine to the smoke from either weak or strong cigarettes for 10 minute periods, 4 times a day from the 3rd to 14th (2nd last) day of pregnancy. Control hamsters were either similarly restrained in a Walton machine equipped with an unlit cigarette, or were not placed in the machine or restrained. Examination of the progeny in the first 6 days of life showed changes in density indices of grouped pulmonary neuroendocrine (NE) cells (neuroepithelial bodies, NEB) that were related to in utero exposure to maternal smoking. Argyrophil NEB were more numerous, larger, and contained more cells at birth among neonates whose mothers smoked the strong cigarette (2.45 mg nicotine and 36.8 mg tar) during pregnancy. This suggests a doserelated effect as the weak cigarette (0.37 mg nicotine and 33.8 mg tar) group did not show such changes. However, some of the changes described did not last through 3 or 6 days of age. The stress resulting from restraint alone also appeared to increase argyrophil NEB indices. Lung tissue volume fraction was increased in the weak cigarette group over all other groups at birth and 3 days; this suggests that low nicotine has the strongest pharmacological effect on lung tissue growth. The medial thickness of pulmonary arterioles was unchanged by either treatment; this provides morphometric evidence that chronic pulmonary hypertension was not present. We could not determine whether the increased NEB indices were caused by increased stainability, by activation of resident reserve cells, or by actual mitosis.
Open Access
An immunocytochemical method for assaying oestrogen receptors in breast cancers. A comparison with the steroid binding assay
(Murcia : F. Hernández, 1988) Ottestad, Lars; Tveit, Kjell M.; Nesland, Jahn M.; Dybsjordl, Indrejit; Bormer, Ole; Hoie, Johan; Pihl, Alexander
The presence of oestrogen receptors was studied in 105 human breast carcinomas using monoclonal antibodies (Abbott ER-ICA kit). The oestrogen receptors of neoplastic cells were semiquantitatively measured and correlations were made to receptor values determined by a dextran-coated charcoal (DCC) steroid binding assay and to histological grade. Immunoreactive cells were found in about 213 of the tumours. Usually only a fraction of the cells within each tumour were immunoreactive, and the staining intensity varied among different cells. In general, well differentiated tumours had a greater proportion of immunoreactive cells than poorly differentiated ones. In most cases (651 98) a good agreement was found between the ER-ICA method and the DCC assay. However, in 33 cases discrepancies were demonstrated.
Open Access
Previously unreported "hemidesmosomal junctions", between folliculo-stellate cells and pituitary adenoma cells
(Murcia : F. Hernández, 1988) Osamu Tachibana; Tetsumori Yamashima
Thirty-eight non-functioning pituitary adenomas were ultrastructurally investigated with particular attention to the Folliculo-Stellate (FS) cells. A large number of FS cells were found in four cases, one of which disclosed a new type of intercellular junction between FS cells and surrounding adenoma cells. These junctions were characterized by 1) the presence of plasmalemmal attachment plaques only in FS cells, 2) the cytoplasmic filaments assembling in parallel to the attachment plaques, 3) the parallel plasma membranes being separated by the intercellular amorphous material and 4) the intercellular space of approximately 25nm width. They were similar to hemidesmosomes, but were quite different from hemidesmosome-like intercellular specializations which have been described in the normal meninges and human meningiomas. Accordingly, we designated these new junctions as nhemidesmosomal int~ctionsu which appeared to be one of the ultrastructural features characterizing FS cells.
Open Access
Cytotoxic effects of 3-methylindole on alveolar epithelial cells and macrophages: with special reference to microtubular and filamentous assemblies in alveolar type I cells of bovine lung
(Murcia : F. Hernández, 1988) Atwal, Onkar S.; Minhas, Kanwal J.; Perry, Michael S.
The alveolar type I cell is a major permeability barrier between the pulmonary interstitium and alveolar spaces and its thin cytoplasmic processes are greatly susceptible to injury. These cells are often observed to undergo progressive vesiculation, vacuolization and desquamation during 3-methylindole (3MI)-induced acute pulmonary edema after oral administration in goats and cattle. The present study describes proliferation of SER and the presence of polymerized tubulin in the form of microtubules arranged in large bundles shown at ultrastructural level as well as with immunofluorescence staining for tubulin in alveolar type I cells 72 hours after 3MI treatment. Such changes were not seen in pulmonary endothelial cells, alveolar type I1 cells, alveolar macrophages and neutrophils. The possible role of microtubules in alveolar type I cell as a mechanistic support to resist disruption against the forces of interstitial and alveolar edema is compared with alveolar type I1 cells, alveolar macrophages and neutrophils. The latter cells undergo dynamic movements in response to inflammatory stimuli and therefore did not show microtubules in their cytoplasm
Open Access
Effects of isoproterenol on the fine structure of the hamster parathyroid gland
(Murcia : F. Hernández, 1988) Shizuko Shoumura; Yuji Iwasaki; Noriko Ishizaki; Shoichi Emura; Tomo Yamahira; Masaki Ito; Hideo Isono
Ultrastructural changes of the parathyroid glands of isoproterenol-treated golden hamsters were investigated. Many chief cells in the parathyroid glands after 5 and 10 minutes of administration of isoproterenol contain well-developed Golgi complexes and granular endoplasmic reticulum, numerous prosecretory granules, and many secretory granules in the peripheral cytoplasm as compared with the control animals. Many chief cells in the parathyroid glands after 1,3,6 and 12 hours of administration have poorly-developed Golgi complexes, granular endoplasmic reticulum, many secretory granules and numerous lipid droplets as compared with the control animals. The morphology of the parathyroid gland after 30 minutes and 24 hours of administration resembles that of the control animals. It is considered that isoproterenol affects the secretory activity of the parathyroid gland.
Open Access
An ultrastructural stereologic study of aldosterone secreting adrenal adenomas and of adjacent zona glomerulosa
(Murcia : F. Hernández, 1988) Belloni, A. S.; Mantero, Franco; Rocco, Stefano; Gottardo, Giuseppe; Mazzocchi, G.; Nussdorfer, G.G.
The ultrastructure of four aldosterone secretine adenomas and of the adiacent zona glomerzosa has been described by "the use of stereological techniques. Adenomatous cells (about 2800 pm' in volume) invariably displayed a striking abundance of lipid droplets. which occupied about 30% of the cytoplasm. Mitochondria prevalently contained tubulo-lamellar or lamellar cristae, hut some cells exhibited organelles with vesicular cristae. Smooth endoplasmic reticulurn (SER) was not very abundant. Small lipofuscin-pigment granules were frequently seen and in a few cells they were exceedingly numerous. Zona glomerulosa cells were smaller (about 950 pm3 in \~olume) and possessed mitochondria with typical tubulo-lamellar cristae, a plentiful SER and few lipid droplets. They showed the ultrastructural features of elements actively engaged in steroid synthesis. The possible origin of aldosteronoma cells from the zona glomerulosa is discussed.
Open Access
Production and immunohistochemical characterization of monoclonal antibodies directed against renal basement membranes of rats
(Murcia : F. Hernández, 1988) Hinsch, Klaus-Dieter; Hansen, Diethelm; Zimmermann, Astrid; Bruchhausen, Franz V.
Basement membranes were separated from rat glomeruli and purified by mild procedures, which led to a highly enriched basement membrane fraction. Here, the production and characterization of five monoclonal antibodies against tubular and glomerular basement membranes are described. These antibodies were analyzed immunohistochemically on frozen sections of rat, bovine, and human kidneys as well as on rat embryos. One monoclonal antibody (BM 0 11) exclusively recognized the glomerular basement membranes, another one (BM 0 VII) bound to tubular basement membranes and to Bowman's capsule. Three antibodies (BM 0 IV, BM M 11, BM M 111) recognized their antigens in both glomerular and tubular basement membranes as well as in mesangial cells. The BM 0 I1 antibody showed a stringent species specificity and bound only to glomerular basement membranes of the rat. The other four antibodies cross-reacted with human and bovine glomerular basement membrane and mesangial antigens; they also bound to other tissues in the developing rat embryo. Antibody binding to specific purified components of the basement membranes such as collagen type IV, laminin, heparan sulphate proteoglycan, and fibronectin was investigated by enzyme-linked immunosorbent assay (ELISA). None of these antibodies reacted with any of these known basement membrane components, indicating that the antibodies may serve as useful tools in future investigations of so far unidentified components of basement membranes.

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