Histology and histopathology Vol.10, nº 1 (1995)

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Open Access
Epithelial lung cell marker, current tools for cell typing
(Murcia : F. Hernández, 1994) Kasper, M.; Singh, G.
This review discusses current irnmunohistochemical and lectin histochemical approaches to identify and to distinguish the different epithelial cell populations of the pulmonary tissue. Special emphasis is given to the characterization of pulmonary alveolar and bronchial epithelial cells and mesothelial cells. Structural proteins, membrane molecules and secretory products of the alveolar epithelium, which have already been characterized and which may be useful for monitoring the developmental or pathological processes, are listed and briefly described.
Open Access
Histochemical and ultrastructural features
(Murcia : F. Hernández, 1995) Fekete, Eva; Resch, B.A.; Benedeczky, I.
The developing enteric nervous systern of the hurnan foetus has been analyzed at the 10th and 18th week of gestation with a special reference to the development of nerve-rnuscle contacts. The rnyenteric plexus forrnation was analyzed by rneans of electron microscopy and on whole-mounts after NADH diaphorase histochernistry. The developrnent of noradrenergic innervation as an extrinsic inhibitory supply was followed by means of a glyoxylic acidinduced fluorescence rnethod. Differentiated neurons and neuroblasts both occurred in rnyenteric ganglia of the 10- and 18-week-old foetus although the ganglionic neuropil was almost unidentifiable ultrastructurally at the 10th week of gestation but was rnature looking at the 18th week. The nerve plexuses connecting the ganglia frequently forrned distant and close rnyoneural contacts. NADH-diaphorase histochernistry revealed that in the 18-week-old hurnan foetus rnost of the neural perikarya was within the ganglia. Also, at the 18th week of gestation a well-defined fluorescent network was dernonstrated within the ganglia, as well as in the internodal segrnents. On the basis of these observations we concluded that the time between the 10th and 18th week of gestation has pararnount irnportance for both the rnorphological and functional rnaturation of the enteric nervous systern.
Open Access
Lectin-binding sites in the epithelium of normal
(Murcia : F. Hernández, 1995) Brinck, U.; Bosbach, R.; Korabiowska, M.; Schauer, A.; Gabius, H.J.
By using histochemical methods, the binding pattern of various lectins in the epithelium of normal human appendix vermiformis was assessed. In addition to plant and invertebrate sugar receptors with nominal monosaccharide specificity for a-L-Fuc (UEA-1), a-DMan and a-D-Gluc (Con A), a-D-GalNAc (DBA), DGalNAc (SBA, HPA) B-D-Gal (RCA-I) and D-Gal (VAA), a mammalian B-galactoside-specific lectin (MW, 14 kDa) was included in the applied panel. The apical surface of enterocytes presented binding sites for RCA-1 on al1 cells, binding sites of UEA-1, DBA, SBA, HPA and VAA heterogeneously and no binding sites of Con A and 14 kDa. Binding sites of DBA, SBA, HPA, VAA and RCA-1 within enterocytes were located primarily focally in a supranuclear position, whereas Con A and 14 kDa bound to the cytoplasm both in apical and basa1 cell parts. In the follicle-associated epithelium more enterocytes expressed SBA- and VAA-binding sites than in the crypt epithelium. No differences between the lectin-binding pattern of M-cells and enterocytes were found in the follicle-associated epithelium. Intraepithelial macrophages were heterogeneously positive for the full panel of applied lectins. In contrast, intraepithelial lymphatic cells expressed binding sites only for RCA-1 and less prominently for Con A, VAA and 14 kDa. Goblet cell mucus contained lectin-binding sites in a heterogeneous manner: binding sites for Con A were not detected in goblet cells for DBA, SBA, VAA and 14 kDa in less than 20%, for UEA-1 in 20-40%, for HPA in 40-60% and for RCA-1 in 60-100% of the goblet cells. Secreted mucus differed in its lectin-binding capacity from intracellular goblet cell mucus selectively by an increase of UEA-1, SBA- and RCA-1-binding sites and a lack of 14 kDa-binding sites. Comparative study of lectin binding to goblet cell mucin in another region of the large intestine, namely the rectosigmoid, demonstrated that DBA, SBA and 14 kDa bound mainly to the dista1 colon, while UEA-1 and VAA labelling was selectively found in Oífprint reguests lo: Dr. Ulrich Brinck, M.D., Department of Pathology, University of Gottingen, Robert-Koch-Str. 40, D-37075 Gottingen, Germany apendiceal goblet cell mucin. Comparing the lectin-binding pattern in normal appendix epithelium and in appendicitis, the percentage of goblet cells expressing DBA- and SBA-binding sites in mucus globules was found to be about 4 times higher in appendicitis than in normal appendix. These results demonstrate that the expression of lectin-binding sites in appendiceal goblet mucin is specifically altered in appendicitis, indicating that there are selective changes of glycosylation of mucin in goblet cells mainly of the lower and middle crypt segment. Changes of lectinbinding pattern in appendicitis are discussed in connection with histochernical findings in inflammatory bowel disease.
Open Access
Human IgA nephritis, immunocytochemical evidence
(Murcia : F. Hernández, 1995) Yoshioka, K.; Maki, S.
This overview summarizes recent information concerning the biopathology of mesangial cell proliferation and matrix expansion which constitute fundamental features in human IgA nephritis. The currently available knowledge. mainly stemming for immunohistochemical observation of human materials, experimental investigations with laboratory animals, and mesangial cell culture studies, emphasizes the importance of cell to cell, cell to soluble factors, and cell to matrix interactions. Mesangial cells, activated by cytokines and growth factors, express adhesion molecules, stimulate proliferation both of themselves and neighbouring cells, and synthesize extracellular matrix. Matrix components, in turn, may influence the behaviour and proliferation activity of mesangial cells, or act as a receptor or reservoir for growth factors. Expression of protooncogenes, regulating cell proliferation and apoptosis, by glomerular cells could be associated with persistent cell replication and chronic tissue damage. These disease processes seem to be common to a group of diseases termed chronic inflammatory proliferative disorders.
Open Access
Structure and function of melanocytes, Microscopic
(Murcia : F. Hernández, 1995) Hirobe, T.
Melanocytes characterized by their tyrosinase activity, melanosomes and dendrites locate in the basa1 layer of epidermis and hair bulb in the skin of mice. Melanocytes differentiate from undifferentiated melanoblasts derived from embryonic neural crest. Melanocyte-stimulating hormone plays an important role in the regulation of the differentiation of mouse melanocytes in the epidermis and hair bulb by inducing tyrosinase activity, melanosome formation, transfer of melanosomes and increased dendritogenesis. The proliferative activity of differentiating epidermal melanocytes of newborn mice during the healing of skin wounds is regulated by semidominant genes, suggesting that the genes are involved in regulating the proliferative activity of epidermal melanocytes during differentiation. The morphology and differentiated functions of mouse melanocytes are shown to be influenced by environmental factors such as ultraviolet and ionizing radiations. From the results of serum-free culture of mouse epidermal melanoblasts, basic fibroblast growth factor is shown to stimulate the sustained proliferation of melanoblasts in the presence of dibutyryl adenosine 3 ' 3 - cyclic monophosphate and keratinocytes. In contrast, melanocyte differentiation in serum-free culture is induced by melanocyte-stimulating hormone in the presence of keratinocytes. These results suggest that the structure and function of mouse melanocytes in the epidermis and hair bulb are controlled by both genetic factors and local tissue environment, such as hormones and growth factors.
Open Access
The use of microwave irradiation for immunohistochemistry
(Murcia : F. Hernández, 1995) Vitarelli, E.; Sippelli, G.; Tuccari, G.; Barresi, G.
We developed a rapid immunohistochernical method using a microwave oven in formalin-fixed, paraffin-embedded sections from normal and pathological tissues. The strongest immunoreactivity was obtained for actin, Ca 125, CEA, pan-cytokeratin, chromogranin A, EMA, GFAP, thyroglobulin, kappa and lambda chains. In control tissues, processed with conventional immunocytochemical procedure, the reactivity was found to be qualitatively and quantitatively similar. Dako EPOS kits were also assayed with good staining intensity, shortening the original technique to 16 min. Our microwave immunohistochemical method is simple, rapid and it may be recommended for use in routine laboratories.
Open Access
Lesions in lambs experimentally infected
(Murcia : F. Hernández, 1995) Masot, A.J.; Gázquez, A.; Regodón, S.; Franco, A.; Redondo, E.
An experimental model was designed to characterize lesions in the lung of lambs inoculated with bovine respiratory syncytial virus (BRSV). 25 Merino lambs of both sexes, with a live weight of 17+3 Kg, received an intratracheal inoculation of 20 m1 saline solution containing 1 . 2 6l~o 6 TCIDSOB RSV (strain NMK-7) per ml. Lambs were laughtered 1, 3, 7, 11 and 15 postinoculation days (PID), and histopathological, immunohistochemical and electron microscopic studies were performed. Results reflected a series of lesions, the most noteworthy of which were bronchiolitis obliterants with destruction of the mucociliary apparatus, the presence of syncytial cells in alveoli and a progressive interstitial reaction. BRSV antigen was detected in lung samples. These changes might be expected to decrease the efficiency of respiratory tract defence mechanisms, rendering the lung parenchyma susceptible to opportunist bacteria1 infection.
Open Access
Giant cell tumor of the ovary. lmmunohistochemical
(Murcia : F. Hernández, 1995) Franco, V.; Florena, A.M.; Orlando, E.; Becchina, G.
Giant cell tumor (GCT) of the ovary ic a rare condiiion, fouiid almost invariably in the context of a mucinous iumor and presenting a microscopic picture indistinguishable frorn GCT of bone. We describe a case of GCT in the wall of a serous cyst of the ovary. An iinmunohistocheniicr~l study was performed using a panel of antibodies to epithelial, niesenchymal and leukocyte antigens. Mononuclear arid giant tumor cells were positive for vimentin; CD 68 and LCA were found only in giant cells whereas actin was only found in inononuclear tumor cells. The immunophenotypic profile of the stronial cells of the residual o\!ary was identical to that of riiononuclear tumor cells. The pi-esentcd data suggest that GCT of the ovary is probnbly ri non-neoplastic lesion of the mesenchyinal stronial cells that react against substances of the associated tuinor or cyst.
Open Access
Vessel wall-dependent metabolic pat hways of the
(Murcia : F. Hernández, 1995) Preissner, K.T.; Pötzsch, B.
The integrity of the vessel wall under quiescent conditions as well as its appropriate responsiveness under conditions of stimulation, inflammation or vascular injury is controlled by a number of adhesive interactions involving distinct cellular receptor systems and various multifunctional adhesive ligands. While a number of these extracellular matrix components of the vessel wall are endogenously produced, secreted and deposited, exogenous adhesion proteins may become translocated from the intra- to the extravascular space by virtue of endothelial cell-specific transport systems. Two prominent examples for each metabolic pathway are discussed. Endothelial cellspecific biosynthesis and secretion as well as deposition of multimeric von-Willebrand-factor within intracellular granules (Weibel-Palade bodies) relates to the first possibility of processing, whereas binding of reactive forms of circulating vitronectin to diverse cellular receptors with subsequent extravasation and deposition into the extracellular matrix appears to be characteristic for the second case. In this review the known features of the metabolic routes of both adhesion proteins are discussed and set in perspective to their functional properties. Their localized mode of action in the vessel wall appears to be crucial for balanced haemostasis and immune systems, two major defence mechanisms of the organism.
Open Access
Effects of bombesin on the morphology and
(Murcia : F. Hernández, 1995) Malendowicz, L.K.; Nussdorfer, G.G.; Miskowiak, B.; Majchrzak, M.
The acute and chronic effects of bornbesin (BM) on the structure and function of rat adrenal cortex were investigated by rnorphornetric and radioirnrnunological techniques. An intraperitoneal bolus injection of 2 yglrat BM rnarkedly raised plasma corticosterone (B) concentration (PBC). The intraperitoneal BM infusion (1 yg/rat.h-l) for 1, 2 or 4 days evoked a notable increase in the nurnber of adrenocortical cells, without inducing apparent changes in either PBC or B output by adrenal quarters. Since proliferation and expression of specialized functions are rnutually exclusive states of cells, our findings suggest that the conspicuous stirnulation of adrenocortical-cell proliferation evoked by BM infusion may be responsible for the apparent lack of effect of this treatrnent on B secretion.
Open Access
Effects of various antineoplastic agents on-2
(Murcia : F. Hernández, 1995) García Reverte, Juana María; Bernabeu-Esclapez, A.; Muñoz Ramos, J.; Vicente Ortega, Vicente; Canteras Jordana, Manuel
An established human melanoma cell line was treated with severa1 concentrations of three antineoplastic drugs: melphalan (0.01 6, 0.032, 0.16 pn), CCNU (1- (2-chloroethy1)-3-cyclohexyl1-- nitrosourea) (0.04, 0.21, 0.42 pM) and 4-OHA (4-hydroxyanisole) (4.01 x 1 0-4, 1 . 2 0~1 0 -2~.,4 ~ pM), and the effects on cell growth and viability were compared. 24 hours after treatment, 4-OHA ( 1 ~ ~ ~ = 2 . 4 ~ 1 0 - ~ pM) was more cytotoxic than melphalan (ID50=0.016 pM) and CCNU (ID50=0.21 pM). However. after 96 hours exposure, the most effective drug was CCNU (growth rate = -1.277), which caused the death of the culture. This was followed by melphalan (growth rate = - 1.024) and finally 4-OHA (growth rate= -0.69). Similar ultrastructural cell injuries were observed after the use of the three drugs: the dilation of endoplasmic reticulum vesicles and the nuclear membrane; mitochondria swelling; and the existence of lamellar structures and cytoplasmic vacuoles.
Open Access
Epithelial-to-mesenchymal change of differentiation.
(Murcia : F. Hernández, 1995) Guarino, M.
ln embryonic morphogenesis, dramatic changes from one state of differentiation to another take place, and some epithelia transform into mesenchymal cells endowed with the ability to migrate and to form connective tissue. In vitro model systems have been developed which have provided new insights into crucial aspects of this differentiation change. Triggered perhaps by either extracellular matrix or growth factors. this phenotypic conversion involves a reorganization of the cytoskeleton, and changes in both cell-cell and cellmatrix interactions. As embryonic and adult tissues contain the same, albeit differently expressed, genetic information, one could expect, under particular circumstances, conversion to mesenchyme from epithelium to occur in adult tissues too. Indeed, there is evidence that this change really occurs in human diseases: some tissue reactions to injury: the process of tumour invasion and metastasis; and the development of carcinosarcomas, are al1 pathological conditions in which an epithelial conversion into mesenchyme probably plays a role. Here, recent observations on embryonic and in vitro epithelialmesenchymal conversion are reviewed. and these data are compared with findings from some pathological situations. Many similarities emerge which further strengthen the belief that this change in differentiation is involved in the pathogenesis, and underlies the pathological pattern of some diseases.
Open Access
The role of xanthine oxidase in
(Murcia : F. Hernández, 1995) Frederiks, W.M.; Bosch, K.S.
Oxygen radicals have been proposed to be involved in the induction of liver cell damage during reperfusion after ischemia. The role of xanthine oxidase in this process and the potential of the antioxidant system have been studied in a model of in vivo ischemia of rat liver followed by 1 h reperfusion by the use of enzyme histochemistry. Based on decreased lactate dehydrogenase activity in certain areas of liver parenchyma, cell damage could already be detected at 1 h reperfusion after ischemia. Incubations performed on serial sections showed that the same areas contained decreased activities of xanthine oxidoreductase, xanthine oxidase, catalase and glucose-6-phosphate dehydrogenase. Some individual cells in the undamaged liver parenchyma expressed a very high glucose-6-phosphate dehydrogenase, which suggests that these cells have a good defence against oxidative stress. It is concluded that oxygen radicals derived from xanthine oxidase do not play a decisive role in the induction of cell damage immediately at reperfusion after ischemia. However, it cannot be excluded that xanthine oxidase present in the blood stream can give rise to the development of additional damage later on.
Open Access
Sequential expression of glutathione-S-transferase
(Murcia : F. Hernández, 1995) Wollina, Uwe; Paus, R.; Feldrappe, S.
The hair follicle is a stnicture showing a very unique cycling of its epithelial components. It is innervated by an abundance of peptidergic nerves, and neuroendocrine cells occur within the epithelium or in close proximity to it. Recently, it has been suggested that hair growth may be regulated by neuropeptides. We therefore investigated the relationship of VIP receptor expression to intracellular phosphotyrosinase and caldesmon involved in the control of growth and differentiation and metabolizing isoenzymes of the glutathione-S-transferase (GST) family alpha, mu, and pi during induced anagen of C57 BI-6 mice by immunohistotochemistry. It was demonstrated that GST isoenzymes were expressed sequentially in the bulge area and the inner hair root sheath as well as in the sebaceous gland epithelium. Caldesmon was present during the early anagen phases within the bulge region, as was phosphotyrosinase. However, phosphotyrosinase expression decreased in late anagen, and recovered again in post-epilation telogen. VIP receptor was expressed within the bulge area during anagen V, but was absent during the other cycle phases. These results suggest a relationship of protein expression to hair cycle phases and in particular a physiological function of VIPIVIP receptor in terminating the extensive hair follicle growth during anagen 1 to IV.
Open Access
Structural organization of enteric
(Murcia : F. Hernández, 1995) Ibba-Manneschi, L.; Martini, M.; Zecchi-Orlandini, S.; Faussone-Pellegrini, M.S
The organization of the Enteric Nervous System (ENS) was studied in the human colon. Fragments of the whole colonic wall were either routinely processed or Zinc-Iodide Osmium impregnated. Single-layer preparations were also obtained from some of the Zinc-Iodide Osmium-impregnated specimens. The results showed some differences in the organization of human colonic ENS from that of other mammals. In fact, the human submucous plexus was made up of three interconnected ganglionated networks arranged along three different planes. With respect to the myenteric plexus, its ganglia were large sized and irregularly shaped. Moreover, during the microdissection of the colonic wall, we found the absence of a cleavage plane between the circular and longitudinal muscle layers; on the other hand the cleavage plane between mucosa and submucosa was not immediately below the muscularis mucosae, but slightly deeper, since the innermost part of the submucosa remained adhering to overlying layers.
Open Access
Localization of NADPH-diaphorase activity
(Murcia : F. Hernández, 1995) Tay, S.S.W.
NADPH-diaphorase activity was localized in pancreatic ganglia of the young chick. At 1 day posthatching, 60% of the neurons in the pancreatic ganglia were NADPH-diaphorase positive. In each neuron, the NADPH-diaphorase labelling was localized mainly in the cytoplasm of the cell body and its proximal processes, but not in the cell nucleus. There was a gradation in the labelling for the enzyme, with some neurons being heavily labelled while others were lightly to moderately labelled. At 7 days post-hatching, 100% of the pancreatic neurons showed NADPH-diaphorase activity and the average size of the NADPH-diaphorase positive neurons had also increased. By 14 days posthatching, al1 the neurons present were heavily labelled for NADPH-diaphorase activity. Some of the labelled nerve processes traversed long distances and finally terminated on other ganglia as well as on the exocrine acinar or endocrine cells. It is concluded that this increase in NADPH-diaphoraseINOS activity in the pancreatic neurons is possibly correlated to the increase in modulation of neurotransmission in the young chick.
Open Access
Peptidergic innervation in
(Murcia : F. Hernández, 1995) Kusakabe, T.; Tawakami, T.; Takenaka, T.
The amphibian carotid labyrinth, which corresponds to the mammalian carotid body and carotid sinus, is innervated by nerve fibers containing substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY). FMRFamide (FMRF). and somatostatin (SOM). SP, CGRP, VIP, and NPY immunoreactive varicose fibers are more densely distributed in the peripheral portion of the carotid labyrinth than FMRF and SOM fibers. The time of appearance of SP, CGRP. and VIP is different for each. First CGRP fibers, then SP fibers appear at an early stage of larva1 development, and finally VIP fibres are detected at a later stage of larva1 development. Most SP fibres show coexistence with CGRP, and some SP fibres which show coexistence with NPY immunoreactivity are assumed to be continuous with those demonstrating VIP immunoreactivity. This indicates the possibility of coexistence of four different peptides in the same nerve fibers within the labyrinth. In various vasculatures of mammals, it has been shown that SP, CGRP, VIP, and NPY have a vasoactive nature in relation to the vascular smooth muscle cells. On this basis, it seems that the target of the peptidergic innervation in the amphibian carotid labyrinth is the smooth muscle cells which are abundantly distributed in the intervascular stroma. Accordingly, the peptidergic innervation may be involved in the vascular regulatory function of the labyrinth, although the possibility that these peptides participate in the chemoreception cannot be ruled out. In addition, the vascular regulatory function of the labyrinth may be modulated by the interaction of multiple neuropeptides.
Open Access
Demyelinating hereditary neuropathies in
(Murcia : F. Hernández, 1995) Guzzetta, F.; Guzzetta, A.; Rodríguez, J.; Deodato, M.; Ferriere, G.
Twenty-three cases of hereditary demyelinating neuropathies are reported, 13 with different types of hereditary motor and sensory neuropathy (HMSN) and 9 with globoid cell or meta-chromatic leucodystrophies. Ultrastructural and morpho-metric studies showed some critica1 pathological features emphasizing: 1) the variability of the recessive forms of HMSN; 2) the morphological distinction between HMSN type 1 and type 111; and 3) differences between these types of HMSN and other «onion bulb» neuropathies such as those found in leucodystrophies, which account for distinct underlying mechanisms.
Open Access
In vitro modifications in the proliferation
(Murcia : F. Hernández, 1995) Carretero, J.; Rubio, M.; Navarro, N.; Prieto, P.; Vázquez, R.J.; Sanchez, F.; Vazquez, R.
In order to establish the correlation between in vitro proliferation rate and morphometric variations of prolactin immunoreactive cells, a morphometric study was carried out in rat pituitary monolayer cultures by means of the double immunocytochemical staining methods employing mouse monoclonal antiproliferative cell nuclear antigen (PCNA) and rabbit anti-prolactin (PRL) as primary antibodies. PCNA was found to be an adequeate marker for proliferation in pituitary monolayer cultures. 48.35+2.78% of the cells present in the culture were in active cell cycle after 3 days of incubation and a similar proportion, 54.93&2.83% was found after 7 days. On the 3rd day, PRL imrnunopositive cells accounted for 15.16*0.21% of the total cellular content in the dishes and 8.68*0.12% of the PCNA immunoreactive cells were also PRL immunopositive cells and, 60.95112.65% of PRL cells stained for PRL and PCNA. On the 7th day, an increase to 32.18f 0.60% of PRL cells was found; the PCNA and PRL cells accounted for 60.32*2.34% of the total PRL cells, and 19.88I1.09% of the PCNA reactive cells stained for PRL. Additionally, the morphometric analysis performed after 3 or 7 days of incubation showed that, while the size of PRL cells remained unmodified, the nuclear area had increased on the 7th day in relation to the 3rd day (pc0.01). These results suggest: 1) PCNA is a valid proliferative marker for pituitary cells in cultures; 2) a very high percentage of the PRL cells was in early proliferation; 3) on the 7th day of incubation, the proliferative rate of PRL cells was very similar to that observed on the 3rd day, suggesting a maintained proliferation for PRL cells at early incubation phases; and 4) the cellular activity, expressed as variations in the nuclear size, was higher on the 7th day than on the 3rd day; in addition, the numerical density of PRL cells increased.
Open Access
Hepatocyte-like cells in the pancreatic islets, study
(Murcia : F. Hernández, 1995) Tsanadis, G.; Kotoulas, Othon B.; Lollis, D.
Objective: To determine the existence of hepatocyte-like cells in the human foetal pancreas. Study design: Foetal pancreas was examined in parallel with two experimental models involving pancreatic tissue regeneration. Foetal pancreases (n=20; 10 to 18 weeks) were obtained from spontaneous abortions and were histologically examined using haematoxylin-eosin and PAS staining. Tissues from food-deprived and copperdeficient female Wistar rats were studied following Dpenicillamine administration, and tissue from female hamsters was evaluated following administration of a -- pancreatic carcinogen. Histological examination in animal studies included haematoxylin-eosin staining. and diaminobenzidine histochemistry. Results: The presence of a characteristic cell-type whose morphology was distinct from islets cells and exocrine pancreas cells was observed in human foetal pancreatic islets. These c ~ l l sw ere morphologically similar to hepatocyte-like cells and were compatible to those observed in the experimental models. Topographical relationships suggest that these originate from stem cells which are related to the pancreas duct cells. Conclusion: We conclude that hepatocyte-like cells or precursors exist in the human foetal pancreas.