Histology and histopathology Vol. 4, nº 4 (1989)
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- PublicationOpen AccessThe phylogenetic odyssey of the erythrocyte. I Hemoglobin: the universal respiratory pigment(Murcia : F. Hernández, 1989) Glomski, Chester A.; Tamburlin, JudithHemoglobin is a molecular entity that is capable of reversibly binding and releasing oxygen in either extra- or intracellular milieus. It is present in scattered invertebrates in physical solution or in cellular sites while in vertebrates it is universally located in circulating erythrocytes. These cells serve as the vehicle for and otherwise foster the optimum utilization hemoglobin. Hernoglobin's variable sphere of respiratory activities can be viewed as reflecting the specific requirements for each organism in which it is observed. Once these concepts have been established and the advantages and limitations of its cytologic packaging recognized, the study of the erythrocyte as expressed in its dimensions, colligative aspects, geometry, internal morphology and pathologic variations can be approached in a purposeful manner.
- PublicationOpen AccessUltrastructural and immunoelectron microscopic studies on infiltrating mononuclear cells in lymphocytic submandibulitis in NOD mice(Murcia : F. Hernández, 1989) Takahide Sugihara; Yasuro Yoshimura; Tanaka, OsamuThe ultrastructural relations of the infiltrating mononuclear cells to the parenchyma1 tissues were studied in the submandibular gland of the female nonobese diabetic (NOD) mouse. In addition, the phenotype of mononuclear cells infiltrating the submandibular , gland has been determined by light and electron microscopy by using monoclonal antibodies against T-cell subsets (Thyl.2, Lytl, Lyt2). Ultrastructurally, lymphoid cells were frequently observed around and in the acini and ducts. Some of the lymphoid cells observed in the acini and ducts were irregular in shape and sometimes sent spike-like projections into acinar and ductal cells. Immunohistochemical study demonstrated that Thyl.2+ cells were predominant among the infiltrating cells, and the majority of these infiltrating T-cells were composed of Lytl+ cells with a small proportion of Lyt2+ cells. By immunoelectron microscopy, lymphocytes carrying Thyl.2, Lytl or Lyt2 antigen were identified, as is demonstrated by an electron-dense reaction product on the entire cell surface, and these immunopositive cells were frequently observed around and in the acini and ducts. Some of the Thyl.2+, Lytl+ or Lyt2+ cells observed in the acini and ducts demonstrated a close contact with acinar and ductal cells and both Lytl+ and Lyt2+ cells sent spikelike projections into them. Occasionally, a partial degeneration of acinar cell adjacent to the invading lymphocytes was observed. These observations suggest that T-lymphocytes are involved in the direct destruction of acinar and ductal cells in the NOD mouse submandibular glands.
- PublicationOpen AccessGlomerular filtration barrier in experimental endotoxin shock, a histopathological and physiopathological study(Murcia : F. Hernández, 1989) Redondo, E.; Roncero, V.; Durán, E.; Gázquez, A.In the present work a morphopathological study is carried out of the glomerular filtration barrier in 20 Large-White pigs weighing 20 kg, subjected to experimental intravenous inoculations of endotoxin from Salmonella enteriditis. The study is completed with the determination of protein plasmatic levels, through urine test with the determination of pH, density, proteins, glucose, ketone body levels and urinary sediment. The histopathological and physiopathological results reveal alterations at the level of the filtration barrier with quantitative differences between the different experimental groups.
- PublicationOpen AccessPeripheral T-cell lymphoma or Hodg kin's disease in a HIV seropositive patient: a histopathological study(Murcia : F. Hernández, 1989) Kanavaros, Panagiotis; Nemeth, J.; Lavergne, Anne; Ngo Van, P.; Galian, AnnieMalignant lymphomas occurring in patients with AIDS are usually derived from the B-cell lineage while T-cell malignant lymphomas are very rare in these patients. We report a HIV seropositive 29-year-old homosexual man in whom cervical lymph node biopsy showed an atypical lymphoproliferative process. On morphological and paraffin section immunohistochemical grounds the possibility of Hodgkin's disease (HD) mixed cellularity was initially suggested, but frozen section immunohistochemical studies revealed that the cellular infiltrate exhibited an aberrant pan T immunophenotype and consequently the diagnosis of peripheral T-malignant lymphomas (T-ML) was made. However, genotypic studies would be required to definitely confirm this diagnosis, in such cases. In our case, varying numbers of small and mediumsized cells were positive for both Leu 3lCD4 and Leu 2lCD8 whereas some large cells reacted only with Leu 3lCD4 antibody. Some medium-sized, large and giant cells showed cytoplasmic positivity for Leu MlICD15. Furthermore, the positivity of many large and giant cells with the activation markers BerH2lCD30, Ki-1lCD30, TacICD25 and HLA-DR suggested an activation state for these cells. Our findings emphasize the usefulness of frozen section immunohistochemical methods in order to investigate the spectrum of lymphoid malignancies occurring in HIV seropositive patients, and confirm results of previous studies which stressed the diagnostic difficulties that may appear in distiguishing HD from peripheral T-ML.
- PublicationOpen AccessEpidermal Langerhans cells in infectious diseases(Murcia : F. Hernández, 1989) Tapia, F.J.; Cáceres-Dittmar, G.; Acuña, Lourdes; Mosca, Walter
- PublicationOpen AccessMuscle regeneration induced by snake venom. A histological and histochemical study(Murcia : F. Hernández, 1989) Peña, J.; Jimena, I.; Luque, E.; Martín, J.D.; Vaamonde, R.This report describes the regeneration pattern of anterior tibial muscle of the rat after the inoculation of the snake venom of Bothrops jararacussu . The results show that this regeneration pattern is rather similar to the pattern described in other experimental models. Three days after the injection, three differentiated areas are established: a periferic one of surviving fibres, a second one called myogenic area, and the last one, more internal, made of necrotic fibres that are phagocited by macrophages. The surface of the surviving muscle fibres has myoblasts sticking to it and five days after, the myogenic area is occupied by many of them. Both the previous phagocytosis and the myoblasts came from the area of current uninjured fibres. After 30 and 60 days the regeneration is completed and there are only a few marks that show that the regeneration has taken place.
- PublicationOpen AccessThe effects of CDP-choline on newborn rat pups with experimental alcohol fetopathy. A Golgi study(Murcia : F. Hernández, 1989) Patt, Stephan; Cervós-Navarro, J.; Stoltenburg-Didinger, G.; Schreiner, C.Generally accepted features of alcoholic fetopathy are delayed maturation and retarded dendritic development of neocortex, hippocampus and cerebellum. The present study investigates the effects of a membrane stabilizing agent (CDP-choline) on Purkinje cells of chronically alcohol intoxicated newborn rat pups, employing a Golgi impregnation technique. Both quantitative and qualitative data indicate that CDP-choline modifies the alcohol induced lesion.
- PublicationOpen AccessUltrastructural and hormonal metabolic studies of rat liver maintained in vitro by perfusion at 30° C and 37O C: a time course study by TEM, SEM and RIA(Murcia : F. Hernández, 1989) Hassan, Ibrahim M.; Al-Ali, Saad Y.; Hassan, MithalIsolated rat liver perfusion system has been extensively used for metabolic and functional studies. Results derived from the application of this system may reflect true biochemical changes but they may also be associated with some structural changes. This study was undertaken to correlate the cytological changes and functional integrity of isolated rat liver perfused in vitro at normal physiological temperature (37°C) and 30°C, using a non-recirculating system. The livers were perfused for 3 hours with modified Ham's F10 culture medium supplemented with thyroxine hormone (T4). The hepatocyte structural integrity was studied by light microscopy, transmission and scanning electron microscopy. The triiodothyronine (T3) and T4 hormones in the perfusion medium and the effluent fractions were assessed by radioimmunoassay. The livers perfused at 30°C remained morphologically intact at the ultrastructural level for 3 hours whilst at 37"C, hepatocytes in the centrilobular zone exhibited marked structural alterations. The percentage of T4 uptake was significantly higher (P < 0.01) in livers perfused at 30°C (50.8 * 7.7% vs 38 * 7.7%, 37"C), but the net T3 output (3.16 * 1.04 pg) and the conversion of T4 to T3 (4 * 0.62%) were significantly higher (P < 0.001) in livers perfused at 37°C in comparison to livers perfused at 30°C (1.61 * 0.84 pg and 1.68 2 0.76%, respectively). In conclusion, at 30°C the hepatic T4 uptake is not inhibited, but the rate of T4 to T3 conversion has decreased, additionally the livers remain morphologically well preserved throughout the experimental period. At 37"C, although T4 to T3 conversion is higher, structurally the livers could not be maintained intact for more than 2 hours. Therefore, isolated rat livers perfused in vitro at 30°C offer the best compromise for further morphological and metabolic studies.
- PublicationOpen AccessNucleolar organiser regions in colonic dysplasia. A preliminary study(Murcia : F. Hernández, 1989) Sutherland, C.G.G.; McMeekin, W.As a preliminary investigation in the evaluation of the argyrophilic nucleolar organiser region (AgNOR) technique in colonic dysplasia, quantitation of AgNORs was carried out in biopsies of normal rectal mucosa and tubulovillous adenomas. The AgNOR counts in the lower third of the normal crypts were approximately twice those in the surface mucosa but there was no significant difference between counts in normal crypt bases and adenomas. It is concluded that the AgNOR technique is unlikely to be of value in the assessment of colonic dysplasia.
- PublicationOpen AccessReorganization of the subplasmalemmal cytoskeleton in association with exocytosis in rat mast cells(Murcia : F. Hernández, 1989) Holm Nielsen, E.; Braun, K.; Johansen, T.The subplasmalernmal cytoskeleton in mast cells has been studied by scanning electron microscopy of the internal side of the plasma membrane. Rearrangement of the dense subplasmalemmal network of actin filaments took place following cell activation by compound 48/80 and secretion of histamine. The rearrangement was a withdrawal of the subplasmalemmal cytoskeleton from the exocytotic sites and the development of bare, filament-free areas around the sites. In calciumdepleted mast cells we demonstrated a dense network that was difficult to break. Activation of the calciumdepleted cells by compound 48/80 did not induce rearrangement of the network, and in parallel there was no secretion of histamine.
- PublicationOpen AccessCompensatory adrenal growth in aldosterone-treated male and female hamsters(Murcia : F. Hernández, 1989) Kasprzak, A.; Malendowicz, L.K.The aim of the study was to investigate the compensatory adrenal growth in aldosterone-treated male and female hamsters. Hemiadrenalectomised and sham-operated animals were treated for 5 days with a daily d-aldosterone dose of 25 pglanimal. In both male and female aldosterone-treated hamsters monoadrenalectomy did not change the relative adrenal weight if compared with sham-operated groups. The fasciculata zonae of monoadrenalectomised aldosterone- treated males was larger and contained more parenchyma1 cells than in appropriate control group. There was no difference in the volume of adrenocortical zones, average cell volume and in cell number between sham-operated and unilaterally adrenalectomised females. In vitro 3H-thymidine incorporation per adrenal was markedly higher in monoadrenalectomised than in sham-operated aldosterone-treated males while the opposite was true for female hamsters. Thus, the action of aldosterone on CAG in the hamster seems to depend on sex, with no effect in males and inhibitory action in females.
- PublicationOpen AccessMorphological and biochemical alterations in growing rats induced by etretinate(Murcia : F. Hernández, 1989) Amo Bernal, C.; Mendez Sanchez, A.; Sinisterra Gago, J.V.; Sierra Plana, M.A.; Fuentes Garcia, A.Etretinate is an aromatic retinoid extensively used on Dermatology. Its toxic effects, however, reduce its application from a clinical point of view. In the present paper, we study etretinate intoxication of 48 growing Wistar rats. The intoxication was for 12 weeks using etretinate doses of 0.5 and 6 (mglkg) / day. The concentrations of etretinate in plasma and liver were determined. Total seric cholesterol and triglyceride concentrations were analyzed. Structural and ultrastructural histological studies of liver samples were carried out. Continuous etretinate ingestions seem to produce an alteration in the detoxication of enzymatic complexes in the growing rats with both the concentrations, due to the increase in etretinate blood plasma observed during the study. There is a relationship between the etretinate dose and its blood plasma concentration and toxic effect, but there is not with etretinate concentration in the liver. The blood plasma concentration of cholesterol and triglycerides is not related to histological liver lesions. The histological study confirms hepatotoxicity with both doses. Nevertheless, the anatomopathological lesions observed do not seem to be related to the blood plasma and liver etretinate concentrations.
- PublicationOpen AccessThe autonomic innervation of the liver and gallbladder of Rana ridibunda(Murcia : F. Hernández, 1989) Azanza, M.J.; Aisa, J.; Junquera, C.; Castiella, T.1 .- The innervation of the liver and gallbladder of Rana ridibunda has been studied by the following methods: (a) demonstration of cholinesterase activity; (b) FIF method for catecholamines; (c) immunohistochemistry for VIP and (d) electron microscopy. 2.- The hepatocytes are arranged in regular rows of hepatic cords, very little connective tissue is distributed in the parenchyma, the innervation being restricted to the big branches of blood vessels. 3.- Well defined cholinergic and adrenergic plexuses surround the hepatic arteries, portal veins and biliary ducts. The VIPergic innervation is scarce in the liver but a richly branched plexus spreads in the wall of the gallbladder. 4.- Cholinesterase-positive cells are widely distributed accompanying the nerve trunks of the gallbladder. The innervation distribution is prominent in the portion of the gallbladder next to the hepatic hilus. 5.- A population of melanin-storing cells besides free melanine granules are present in the liver parenchyma and are prominent in the gallbladder where the melanocytes are disposed in close contact with blood vessels and nerve structures. We have observed that the number of these visceral melanocytes considerably increases in winter, particularly in the liver.
- PublicationOpen AccessMorphological evaluation of steatosis in monolayer cultures (MDCK cells) after treatment with gentamicin and valproic acid(Murcia : F. Hernández, 1989) Stiemer, B.An attempt was made to develop a method for the morphological and morphometric evaluation of fat inclusions in monolayer cell cultures. For this purpose we used electron microscopy (SEM and TEM) and Sudan black staining methods followed by morphometric evaluation techniques. MDCK (Madin-Darby-caninekidney) cells were treated with valproic acid (VPA; 10, 100, 1000 pglml medium for 22 hrs) and gentamicin (1, 10,50 pglml for 60 min). Sudan black staining revealed a dose-dependent increase in fat deposits after application of the two substances. SEM inspection showed differences in the size and distribution of the fat inclusions. TEM findings only partly allowed us to differentiate clearly between VPA- and gentamicin-induced fat inclusions.
- PublicationOpen AccessConnective tissue of the livers of newborn and adult marmosets (Callithrix jacchus)(Murcia : F. Hernández, 1989) Daqun, Xu.; Schroter-Kermani, C.; Hinz, N.; Merker, H. J.Immunofluorescence microscopic and electron microscopic investigations revealed components of the matrix and of the basal lamina (collagen type I, 111, IV and V, BL-heparan sulfate and fibronectin) in the sinus wall (Disse's space) of the livers of newborn and adult marmosets (Callithrix jacchus). Collagen type I was missing in both the two age groups. Small amounts of laminin were present in the livers of newborn and absent in those of adult animals, whereas collagen type I11 occurred in the form of delicate fibres. Light microscopic inspection showed a continuous distribution of all other components in the sinus wall. The amount of collagen type I11 and V increased depending on the age. Electron microscopic investigations revealed single or bundled fibrils (20-30 nm) and filaments (10-12 nm). After addition of tannic acid, plaques of a fine-filamentous network and incorporated granules were observed. After addition of resting Ruthenium Red, electron-dense granules (20- 60 nm) were irregularly distributed in the structureless space, resting on collagenous fibrils and cell membranes. The fibrils were allocated to collagen type 111, the filaments to collagen type V. The plaques were supposed to contain heparan sulfate. collagen type IV and fibronectin The absence of a Lamina densa of the basal lamina was attributed to the absence of laminin which probably plays an important role in the formation of this layer. Differences in the distribution pattern of the matrix components and thus a functional mosaic of the permeability of Disse's space were assumed. The complete absence of collagen type I and laminin in the lobules makes the adult marmoset liver especially suited for studies on the importance of this collagen type under pathological conditions, since both components are expressed in this way.
- PublicationOpen AccessPalatal shelf reorientation in hamster embryos following treatment with 5-fluorouracil(Murcia : F. Hernández, 1989) Shah, Ravindra M.; Chen, Yung Ping; Burdett, David N.A study was undertaken to examine the issue of whether achieving a critical mass of cells andor palatal shelf volume during vertical development of shelf is essential for reorientation to occur. In control and 5- fluorouracil (5FU)-treated hamster embryos' palatal shelves, at different times during gestation, the numbers of both epithelial and mesenchymal cells were counted and cross-sectional area was measured. DNA synthesis was measured by 3H-thymidine incorporation and was used as an index of growth by cell proliferation. The control data indicated that, unlike development during initial 24 hours, the later period of vertical palatal development was characterized by a steady level of mesenchymal and epithelial cell numbers and palatal shelf area. Following 5FU treatment all the measurements were reduced, and until they reached the equivalent of control values, the palatal shelves did not reorient. The density of mesenchymal cells in the developing palate did not seem to affect cell number. On the basis of the analysis of results of the present study, along with those reported in the literature, it is suggested that, in hamsters, acquisition and maintenance of both a specified number of mesenchymal cells and shelf area, at least 24 hours prior to reorientation, may be critical for ensuing mesenchymal differentiation to enforce palatal shelf reorientation on schedule. 5FU affected these features to delay reorientation of the palatal shelf.