Histology and histopathology Vol. 5, nº 3 (1990)

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Serotonin-producing pancreatic endocrine tumour. Histological, ultrastructural and immunohistochemical study of a case
(Murcia : F. Hernández, 1990) Kanavaros, Panagiotis; Hoang, Catherine; Le Bodic, Marie Francoise; Polivka, Marc; Hautefeuille, Pierre
Serotonin-producing pancreatic endocrine tumours are rare neoplasms which in most cases exhibit malignant biological behaviour. These tumours, in the majority of the well-documented cases, are composed of argyrophil- and argentaffin-positive cells which contain large pleomorphic neurosecretory granules. In contrast, argyrophilic non-argentaffin pancreatic endocrine tumours with tumour cells containing round neurosecretory granules are exceptional. In this study we describe such a tumour not associated with clinical evidence of carcinoid syndrome in a 60-year-old woman. Histological examination revealed tumour extension in pancreatic lymphatic vessels and veins but no evidence of locoregional or distant metastases. Ten months after surgery the patient showed no recurrence of the disease. Immunohistochernistry revealed cytoplasmic serotonin production in the tumour cells which were negative for anti-gastrin, insulin, glucagon, somatostatin, pancreatic polypeptide (PP), vasoactive intestinal peptide (VIP) and ACTH. This study emphasizes the usefulness of combined ultrastructural and immunohistochemical investigations in order to identify and characterize the rare pancreatic endocrine tumours with serotonin production.
Open Access
The ruputure spleen. A hisological, morphometrical an immunohistochemical study
(Murcia : F. Hernández, 1990) Barnard, H.; Dreef, E.J.; van Krieken, J.H.J.M.
A traumatically ruptured spleen is regarded as a proper control in many histological and immunological studies on the human spleen. This paper compares spleens that ruptured due to trauma and spleens which were removed during surgery in patients without splenic pathology. Based on a histological, morphometrical, and immunohistochemcial description of the control spleens it is shown that the traumatically ruptured spleens contain alterations in the lymphoid tissue. The amount of white pulp is increased due to a larger amount of CD4- positive lymphocytes. Furthermore there are alterations in lymphocyte populations in the different splenic compartments. It is concluded that spleens that rupture may be predisposed due to immunological stimulation.
Open Access
Ground-glass hepatocytes. ligth and electron microscopy. Characterization of the different types
(Murcia : F. Hernández, 1990) Jaime Vázquez, J.
Morphological observations of paraffinembedded histological sections stained with H&E led to the discovery of some cytoplasmic changes which occur in different conditions, although they look alike under the light microscope. These hepatocytic changes consisted basically of homogeneous areas which are weakly eosinophilic in H & E-stained sections. They are frequently referred to as «inclusion» bodies, even when they are not true inclusions. The hepatocytic changes obsewed in HBsAg carriers, in chronic alcoholic patients treated with cyanamide to discourage them from drinking alcohol, in Lafora's disease, and in glycogenosis type IV, look very similar in paraffin sections stained with H&E. Nevertheless, they can be differentiated using ancillary techniques. On electron microscopy they do not look alike. Of particular interest are the «inclusion» bodies induced by cyanamide, a predictable and reproductible lesion, which in man eventually leads to cirrhosis. Other types of hepatocytic changes also giving a rather vague eground-glass» appearance to the cytoplasm are those resulting from intracytoplasmic accumulation of proteins, particularly fibrinogen, and those observed in patients treated with different drugs.
Open Access
Studies with the Golgi method in central gangliogliomas and dysplastic gangliocytoma of the cerebellum (Lhermitte-Duclos disease)
(Murcia : F. Hernández, 1990) Ferrer, I.; Martí, E.; Guionneti, N.; Bella, R.; Serrano, T.; Towse, J.; Conesa, G.; Isamat, F.
The rapid Golgi method, combined with current optical and electronmicroscopica1 techniques, was used in three central gangliogliomas and in one dysplastic gangliocytoma of the cerebellum to study the morphology of ganglionic cells. Gangliogliomas were composed of bipolar, fusiform and radiate cells with dense core and clear vesicles in the perikaryon and cellular processes, the number of each cellular type varying from one case to another. These features, together with the fact that isodendritic neurons are considered to be phylogenetically old neurons, suggest that these tumours are composed of «primitive» neurons that are not homogeneous with regard to their morphology. In contrast, ganglionic cells in dysplastic gangliocytoma are huge cells with long, stereotyped neurites that establish unique asymmetric contacts with neighbouring perikarya and neurites by means of claw-shaped processes covered with synaptic buttons. These morphological characteristics are different from those of any other neuron of the CNS.
Open Access
Effect of ethanol on the cerebellar cortex of the chick embryo
(Murcia : F. Hernández, 1990) Quesada, A.; Prada, F. A.; Espinar, A.; Genís-Gálvez, J. M.
Open Access
Scanning electron microscopic study of natural killer cell-mediated cytotoxicity
(Murcia : F. Hernández, 1990) Fernández-Segura, E.; García, J.M.
The interaction between human natural killer (NK) celis and NK-susceptible target cells, as well as the mechanism involved in target cell lysis, were studied with scanning electron microscopy (SEM). Low density human peripheral blood lymphocytes, highly enriched with large granular lymphocytes (LGL), were used as effector cells, and K562-cells were used as NK-susceptible target ceiis. The surface features of LGLINK cells were examined under SEM. In the area of interaction, NWtarget-cell conjugates showed microvilli andor filipodia, and extensive areas of intercellular contact. In addition, the effector cells in some NWtargetcell conjugates were polarized toward the target cell. Changes in target cell surface features included loss of microvilli, large surface blebs and the appearance of small pore-like lesions on the cell membrane. Our findings show that target cell lysis occurred by apoptosis and plasma membrane lesions analogous to those seen during complement-mediated cytotoxicity.
Open Access
Variations in fibre composition of the gastrocnemius muscle in rats subjected to speed training
(Murcia : F. Hernández, 1990) Morales-López, J. L.; Agüera, E.; Miró, F.; Diz, A.
Thirty-six adult Wistar rats were divided into three groups. One group was used as a control, and the other two underwent different training programmes in which greater relevance was attached to the intensity of exercise than to its duration. Samples of the red and mixed portions of m. gastrocnemius (caputlateralis) were stained with m-ATPase to determine the percentage of type 1, IIA and IIB fibres, and with NADH-TR in order to quantify variations in the percentage of low staining intensity (FG) fibres. The most notable results obtained were: a) the ratio of type 1 type 11 fibres remained unchanged; b) the proportion of IIA fibres increased, while that of IIB fibres decreased correspondingly; c) FG fibres, which were virtually absent from the red portion, recorded a clear decrease which was more marked, and occurred more rapidly, than in IIB fibres. These differences were al1 statistically significant in the mixed portion of the muscle. Adaptative changes in fibre composition in the red portion were less marked.
Open Access
The autonomic innervation of the liver and gallbladder of Podarcis hispanica
(Murcia : F. Hernández, 1990) Azanza, M.J.; Aisa, J.; Junquera, C.
The innervation of the liver and gallbladder of the lizard Podarcis hispanica has been studied by the following methods: a) demonstration of cholinesterase activity; b) FIF method for catecholamines; and c) immunohistochemistry for VIP. The hepatic parenchyma of the reptile's liver show hzpatocytes arranged in regular rows of hepatic cords, the portal triad being typical of higher vertebrates (birds and mammals). Nerve fibers are found in the scarce connective tissue distributed among the hepatocytes. The innervation is restricted to the big branches of blood vessels and biliary ducts. It is represented by cholinergic, noradrenergic and VIPergic fibers. The gallbladder shows a well developed cholinergic plexus with pyramidal cells in the interconnection points of the fiber network. The noradrenergic and VIPergic plexuses are also more widely distributed in the gallbladder thali in the liver.
Open Access
High resolution SEM analysis of acellular glomerular basement membrane following pepsin digestion, intrinsic fibrillar structures
(Murcia : F. Hernández, 1990) Berger, Walter J.; Carlson, E. C.
Microdissection of acellular rat renal cortex with pepsin was carried out to investigate the morphological substructure of glomerular basement membrane (GBM) by high resolution SEM. Renal cortical blocks (< 5 mm ) from adult male Sprague Dawley rats were rendered acellular by sequential detergent extraction and digested up to 184 hrs with 5 mglml pepsin (185 U/ mg) in 0.5 M acetic acid (pH 2) at 10-15°C. Samples were conventionally prepared for SEM, and observed at original magnifications of 500-100,000 diameters. At low magnifications (500-5,00Ox), acellular GBM surfaces appeared smooth at al1 digestion times. At higher magnifications (50,000-100,00Ox), control GBM surfaces were finely granular. Granule diameter ranged from 20-80 nm, with most between 30-40 nm. Pepsin digestion did not affect average granule size. Beginning at 44 hrs of digestion, intrinsic fibrillar structures comprised of linear arrays of 20-40 nm granules were observed onlin GBM surfaces. At later incubation times, this component of GBM became more extensive. At 160 hrs, the fibrillar arrays frequently bifurcated and showed distinctive «forked» termini, some of which comprised two sides of a triangle (120-150 nm on a side). Fork «handles» (310-350 nm in length) radiated from each angle of the triangle. These sometimes terminated in large granules (approximately 100 nm in diameter), two of which appeared to connect fibrillar arrays end-toend. Together with other arrays, the interconnected triangles appeared to comprise a three-dimensional meshwork extending into the GBM and possibly providing support for, its granular components.
Open Access
Human pulmonary dirofilariasis, report of a new European case
(Murcia : F. Hernández, 1990) Fabbretti, G.; Fedeli, F.; Alessi, A.; Boaron, M.; Salpietro, V.; Brisigotti, M.
We report a new European case of pulmonary dirofilariasis occurring in an Italian patient. The paper emphasizes the peculiar pathological features of Pulmonary Dirofilariasis, that, on clinical and radiological grounds, closely irriitates primary or secondary neoplasms. The disease characteristically presents itrelf as a solitary subpleural coin-like lesion, histologically corresponding to a well demarcatec,, roughly spherical infarct, centered by a medium-sized thrombosed artery whose lumen contains the parasite, i.e. a Dirofilaria nematode.
Open Access
Histological features and histochemistry of the mucous glands in ventral skin of the frog (Rana fuscigula)
(Murcia : F. Hernández, 1990) Els, Willem J.; Henneberg, Renata
The glycoconjugate components of secretory granules were analyzed in cells of mucous glands in ventral skin from Rana fuscigula. The analysis was done with standard histochemical methods on semithin glycol methacrylate-embedded tissues. The staining patterns in semithin sections were comparable to those using paraffin-embedded tissue while the cytological detail was better preserved. The mucous glands contained at least two different types of secretory cells lining the lower two-thirds of the mature gland: a principal cell type filled with dense staining secretory granules and a solitary type containing paler staining, globular secretory granules. The principal type of cell contained variable amounts of acid glycoconjugates; predominantly carboxylated but also variably carboxylated and weakly sulfated glycoproteins. Other secretory cells contained mainly neutral glycoproteins. The results indicated that the mucus is a heterogeneous substance and that one cell type may produce different secretory products. We suggested that the variability in histochemical staining might be related to the sequence of biosynthesis of the secretory granule.
Open Access
Ultrastructural characteristics of anterior gut innervation of Gallus gallus
(Murcia : F. Hernández, 1990) Aisa, J.; Parra, P.; Azanza, M.J.
The enteric nervous system of the bird's anterior gut is very well developed. Myelin fibres are seen accompanying the nervous trunks up to the mucous layer. Glial cells duplicate the number of neurons in the myenteric plexuses. Their number decreases at the submucous plexuses, but it is always higher than the neurons. Isolated neurons are widely spread in the circular muscle coat accompanying the nervous trunks which can be inter and intrafascicularly located. Direct synaptic contacts with the soma neurona1 membranes are very often seen. We have never observed synaptic specializations. The most prominent varicosities either in the peripheric nervous trunk axons or directly laying on the soma membranes are those containing peptidergic or mixed vesicles of cholinergic and peptidergic types. The neurons show big nuclei of different size and shape. Neighbouring smooth muscle cells show abundant caveolae near the nervous elements. Although we have not observed close contacts with glands, thin axon bundles spread near the glandular cells of the mucous layer .
Open Access
Vasopressinergic neurons in periependymal and periventricular areas of the rostral third ventricle
(Murcia : F. Hernández, 1990) Roy Choudhury, S.; Ray, P.K.
On the lateral wall of the rostral third ventricle, an area separates the ependyma from the neurosecretory PVN neurons. Since VP from the latter discharges into the vasculature, the above area may be regarded as constituting an interface between the ventncular and vascular compartments of the CNS. As VP release into the two compartments is integrated, the interface region has been explored for possible existence of a neural infrastructure that would allow such an integration. Immunohistochemical staining for VP following colchicine treatment reveals the presence of an elaborate vasopressinergic network in the interface region that is divisible into a media1 periependymal and a lateral periventricular area. A closer examination indicates that the ependymal, periependymal, periventricular and PVN areas (in that order medio-laterally) are al1 interconnected through this network. The media1 area appears to be receptive in nature, while the connectivity of the lateral area points to an effector function. All in all, such a neural network would provide a sound morphological basis for integration of neuroendocrine mechanisms modulating VP release into the ventricular and vascular compartments of the CNS.
Open Access
An ultratructural and immunohistochemical study of extracellular matrix in meningiomas
(Murcia : F. Hernández, 1990) Hisashi Nitta; Tetsumori Yamashima; Junkoh Yamashita; Toshihiko Kubota
Extracellular matrix of meningiomas was studied by light and electron microscopy with the aid of immunohistochemical techniques. Special attention was paid to the distribution of type 1,111, IV, V collagens and laminin with a comparison between meningothelial and fibroblastic types. Connective tissue fibers and basement membrane were not found among the tumor cells in the meningothelial type, but were found in the fibroblastic type. The immunolocalizations were consistently demonstrated extracellularly, but were not within the cytoplasm. Type 1, 111 and V collagens were usually demonstrated in the fibrous septum in the meningothelial type, while they were localized among the tumor cells in the fibroblastic type. Furthermore, type IV collagen and laminin were demonstrated within the vascular walls or around the syncytium in the meningothelial type, while they were localized among the tumor cells in the fibroblastic type. In both types the expression of type IV collagen and laminin was closely related to the distribution of basement membrane. Although meningothelial and fibroblastic meningiomas showed quite different distribution of extracellular matrices, the profile of collagen types expressed by these two basic types was essentially the same. The cellular derivation of meningiomas was discussed with particular attention to the structure of human arachnoid villi and meninges.
Open Access
Effects of photoperiod and temperature on testicular function in amphibians
(Murcia : F. Hernández, 1990) Paniagua, R.; Fraile, Benito; Sáez, F.J.
Most amphibians present an annual testicular cycle characterized by a quiescent period (late autumn-winter) and a spermatogenic period (spring and summer). At the end of the period of spermatogenesis undifferentiated interstitial cells transform into steroid-secreting Leydig cells which regress in spring at the beginning of the new spermatogenetic cycle. The testicular cycle is controlled by the pituitary gonadotropin levels which are high in autumn and winter, low in spring and increase temporarily in the middle of summer. Photoperiod and temperature seem to be the most important externa1 factors involved in the regulation of this cycle in many amphibian species since the colder the geographic area, the longer the quiescent period and the shorter the spermatogenic period. This suggests the occurrence of a potentially continuous cycle in these species, in contrast with that which occurs in other species having an endogenous rhythm of testicular function which is much less sensitive to environmental factors. Although the specific response to temperature can vary widely between species, the most frequent observation in amphibians with a potentiaiiy continuous cycle is that exposure to mild temperatures (15-20° C, according to the spring temperatures of the different geographic areas) stimulates spermatogenesis even during the period of testicular quiescence. If this mild temperature is combined with a long photoperiod, complete spermatogenesis is attained. Experiments performed during the period of germ-cell proliferation (development from spermatogonia to round spermatids) indicated that low temperatures (below 11° C) as well as short photoperiods (less than 8 h of light) hinder germcell proliferation. Moderately high temperatures (about 30° C) do not impair this proliferation. In the newt Offprint reqoests to: Dr. R. Paniagua, Department of Cell Biology and Genetics, University of AlcalA de Henares, E-28871 Alcala de Henares, Madrid, Spain Triturus marmoratus, it has been shown that an excessively long photoperiod (over 16 h) has the same effect as a short photoperiod. In this species eyes are not required for the testicular photoperiodic response. Photoperiod appears to have no effect on spermiogenesis (differentiation of round spermatids into spermatozoa), because once round spermatids are formed, spermiogenesis will occur even in total darkness. Mild temperatures seem to be necessary for spermiogenesis as well as for androgen biosynthesis because neither process will take place at extreme temperatures. Results on the effect of photoperiod in steroidogenesis differ between species.
Open Access
Effect of chronic alcoholism on the human hippocampus
(Murcia : F. Hernández, 1990) Bengochea, O.; Gonzalo, L. M.
The effect of chronic alcoholism on the human hippocampus was studied in 21 patients, divided in 4 groups: Group A under 45 years, group B 46-59 years, group C 60-69, and Group D over 70 years; and compared with age-matched control patients who died without neurological complications. The gyrus d~ntatus and the ammonic fields CA1 through CA4 were analyzed by counting the number of neurons and the size of the nuclear area. Both parameters were evaluated statistically. The most important findings were a high neuronal loss in alcoholics in the first age group. In addition, the hippocampal neurons failed to display a vicarious reaction, since the nuclei did not show any increase in size despite the intense neuronal loss. Our results point out an early neuronal loss in the hippocampus of alcoholic patients higher than agematched controls, as well as a lack of reaction to the neuronal insult .
Open Access
Distribution of serotonin-immunoreactive cells in the mouse pancreas during development
(Murcia : F. Hernández, 1990) Rodríguez Sánchez, F.; Casar, J.; De Castro, J.M.; Del Águila, F.
The distribution and time of appearance of 5HT-storing cells were studied in samples from the pancreas of mice embryos from 7 to 19 days of gestation. Additonally, 1- and 15-day-old newborns and adult specimens were also examined. Serotonin-immunoreactive cells appeared for the fist time in the dorsal pancreatic primordium at 10 days of gestation and increased rapidly in number through E15. From this age, the cellular number disminished in the exocrine pancreatic parenchyma, although an increase of endocrine cells in Wirsung's duct can be detected. At day 15 of extrauterine life, we could only observe some cells in the surface epithelium of pancreatic duct of Wirsung. Islet immunoreactive cells could only be detected in adult animals.

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