Histology and histopathology Vol. 9, nº 1 (1994)

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Open Access
Pulmonary response to bovine albumin. A morphometric study in rats
(Murcia : F. Hernández, 1994) Escolar Castellón, J.de D.; Alfaro, E.; Roche Roche, P.A.; Almajano, C.; Gallego, B.
The following hypothesis is proposed: that hypersensitivity pneumonitis (HP), experimentally induced in rats, is the cause of a thickening in the alveolar wall, a decrease in the size of the alveole, hyperplasia in the bronchus-airociated lymphoid tissue (BALT) and hypertrophy in the goblet cells. Wistar rats were classified into two different groupi, namely, non-treated animals and animals exposed to bovine albumin (BA). A morphometric study was carried out and the following variables were quantified: a) percentage of lymphocytes. neutrophils and alveolar macrophages of the bronchio-alveolar lavage (BAL); b) the interstice of the alveole, the alveolar chord length. the alveolar wall thickness and the number of alveolar macrophages with hemosiderin within its cytoplasm; c) the size of lymphatic area (LA) in BALT, the length of the lymphatic epithelium (LEp) in BALT and the percentage of goblet cells in the bronchial epithelium. The following results were obtained from the animals exposed to BA: 1) a significant increase in both lymphocytes and neutrophils of BAL, and of alveolar macrophages with hemosiderin in its cytoplasm; 2) a significant thickening of the alveolar walls and the BALT elements, which confirms the above mentioned hypothesis; 3) a significant increase in the alveolar chord and a significant decrease in the number of goblet cells of the bronchus, which contradicts the above mentioned hypothesis. The increase in alveolar macrophages with hemoiiderin is related to an increase in the capillary alveole perrneability, which, together with the variations in the BAL formula, lead us to consider the existence of an inflammation in the interstitial alveole; this interstitial alveole inflammation explains the alveolar wall thickening and the BALT hypertrophy.
Open Access
Histopathological characterization of photochemical damage in nervous tissue
(Murcia : F. Hernández, 1994) Van Reempts, J.; Borgers, M.
This paper discusses histological and ultrastructural changes produced by dye-sensitized photoreactions in the central and peripheral nervous system. Particular attention has been given to morphological outcome in experimental models which reproduce widespread clinical pathologies, e.g. stroke, spinal cord injury and peripheral neuropathy. Evaluation of structural alterations may not only help to characterize the evolution of these disease processes but also allow us to study possibilities of therapeutic intervention.
Open Access
coexistence of serotonin and cholecystokinin in paraneurons of the foetal sheep lung
(Murcia : F. Hernández, 1994) Balaguer, L.; Romano, J.; Ruíz-Pesini, P.
The coexistence of serotonin and cholecystokinin was studied in foetal sheep lungs at pseudoglandular stage of developtnerit by light inicroscopic itninunohistochemistry. The coexistence was examined by staining consecutive sections with the different antibodies. Serotonin and cholecystokinin iniinunoreactivity was found within consecutive sections of most bronchopultnonary neuroepithelial bodies and in consecutive sections of the same intrapulmonary autonomic gariglia.
Open Access
The role of the proteasome in cellular protein degradation
(Murcia : F. Hernández, 1994) Driscoll, J.
Eukaryotic cells contain a major intracellular proteolytic activity known as the proteasome. The proteasome is a strongly conserved cylindrical structure of high molecular weight (650 kDa, -20 S) and desmonstrates multiple endopeptidase activities. The general structural, biochemical and genetic features of the proteasonie are conserved from archaebacteria through yeast to humans. This structure fulfills an essential role by functioning as the proteolytic core of a 26 S multienzyme complex responsible for the energydependent degradation of ubiquitinated proteins. The bulk of intracellular proteolysis appears to be through the ubiquitin-dependent pathway. Incorporation of the proteasome into the 26 S niultienzyme complex appears to confer both a specificity for ubiquitinated proteins as well as a nieans to tightly regulate proteolytic activity. Thus, one function of the proteasome is required for the degradation of either abnormal or certain regulatory proteins by the ubiquitin pathway. Proteasoine subunits appear to be encoded by a related gene family as defined by extensive sequence similarities. The gene products are confined to either of two general classes: a-type which appear to be structural and 8-type which may be catalytic. Genes encoding at least two proteasome subunits map to the Ma-jor Histocompatibility Complex. Accurnulating evidence points to the proteasome (or a specialized form) participatirig in the cytosolic degradation of these viral proteins upon cellular infection. Through a previously unforseen mechanisin. it appears that the products from the digestion of the viral proteins may be rescued from further digestion to amino acids and shuttled from the cytoplasm through the endoplasniic reticulum to the cell surface where they serve as antigenic peptides for recognition by the immune system. The proteasome may have been recruited by the immune system to serve as the cytosolic activity responsible for generating these antigenic peptides. The proteasome may function in the ubiquitindependent degradation of not only certain self-proteins but may fulfill a second essential role in the degradation of proteins originating from viral infection
Open Access
DNA synthesis in the embryonic chick lens epithelium ¡S arrested after experimental lens rotation
(Murcia : F. Hernández, 1994) Prada, F. A.; Garcia-Lomas, V.; Genís-Gálvez, J. M.
Using autoradiographic technique, we have studicd DNA synthesis in normal embryonic chick lens epitheliuni and after experimental lens rotation. Analysis of the autoradiograms clearly demonstrates that when the lens priniordium was rotated 180". so that lens epithelium was placed facing the interior of the optic cup. the lens epithelial cells completely stop DNA synthesis. This fact suggests that some retinal and vitieal factors are responsible for differentiation and replicative capacity of the lens epithelial cells.
Open Access
Fine autoradiographical study on scale morphogenesis in the regenerating tail of lizards
(Murcia : F. Hernández, 1994) Alibardi, Lorenzo
Regenerating scales in lizards originate as pockets in the epidermis instead of epidermal elevations as during embryo development. The morphogenesis of scales in the regenerating tail of the lizards. Anolis and Lamphropholis was studied after peritoneal injection of 3~-thymidineT. he tracer was localized in the forming epidermis after progressive post-injection times, by means of autoradiographY on plastic sections. After 4-5 hours post-injection of H-thymidine, the radioactivity was localized in the basal layer. After 2 to 4 days postinjection labelled cells were seen in the basal and intermediate spinosus layers but not in the uppermost keratinizing layers. Labelled cells were seen in the differentiating cornifying layers (pre-B and pre-a) 6-8 days post-injection. At 12-14 days post-injection almost no radioactivity was seen in the basal layer or in the living part of the epidermis. A few labelled cells were present in the dense keratinizing layers of the sloughing wound and interscale lacunar layers. This study shows that scale formation and morphogenesis in the regenerating tail is brought about by a localized cell proliferation along the regenerating epidermis. In the forming scales the percentage of labelled cells in the distal side (future dorsal part of the new scale) is much higher than in the proximal side (the future ventral side of the scale), so that overlapped scales are generated.
Open Access
Mucopolysaccharide histochemistry of the oviduct of the toad, Bufo melanostictus, before and during ovulation
(Murcia : F. Hernández, 1994) Tan, C.K.; Chen, T. W.; Gan, J.M.L.
In both non-ovulating and ovulating toads of the species, Bufo melanostictus, the fimbrium stained only weakly for mucopolysaccharides (MPs) whereas the infundibulum stained strongly for neutral MPs and also for glycogen. In the non-ovulating toad, only neutral and sulphated MPs were detected in the goblet cells of the upper magnum, whereas sulphated, neutral and sialomucins were detected in the glands. In the middle magnum, sulphated and sialic acid-containing carboxylated MPs were detected in both the goblet cells and glands. In the lower magnum, neutral, sulphated, and sialic acid-containing MPs were detected in the goblet cells and only sulphated and sialic acidcontaining MPs were detected in the glands. In the isthmus and ovisac, only sulphated MPs were present in the goblet cells. During ovulation, there was no change in the distribution of sulphated MPs throughout the oviduct. Sialic acid-containing MPs could not be detected in many of the goblet cells of the upper and lower magnum nor in most of the glands of the lower magnum.
Open Access
Topographic distribution and quantitative analysis of retrograde-labelled cells in the substantia nigra following administration of fluoro gold in the caudate putamen nucleus
(Murcia : F. Hernández, 1994) Juanes, J.A.; Bentivoglio, M.; Riesco, J. M.; Chen, S.; Vázquez, R.
The organization of the efferent projections of the substantia nigra following the irijection of 0.4 pl of 2% Fluoro Gold into the caudate-putamen nucleus of rats of both sexes was studied. After a survival period of 77 h. the greatest number of labelled cells was visuali~ed in ihe substantia nigra, with lower numbers in adjacent structures (ventral tegrnental area. zona incei-ta and ~rredial leinniscus). The cclls featured an intensely fluorescent goldcoloured soma and were varied in .shape (round or n\:al); short sinuous projectioris arose out of thein. corresponding to the dendrites and axons of the neurons. ln the statistical study. no significant differences were observed between r-iiales and females. A sirrrilar pattern of distribution was found rangirig Sr0111 ihe rostral to the caudal regions of the substantia niyra. occupying the whole of the extent of the suhstantia nigra; this distribution was most predominant in the rostro-caudal transition of the pars compacta of the substantia nigra.
Open Access
Effects of in utero exposure to low dose ionizing radiation on development in the rat
(Murcia : F. Hernández, 1994) Bruni, J.E.; Persaud, T.V.N.; Froese, G.; Huang, W.
Most studies of in utero effects of ionizing irradiation involve high doses and examination at postnatal intervals. Little information is available on the effects of low levels of ionizing radiation on embryogenesis. The developmental effects of in utero exposure to 50 cGy gamma radiation on gestational day-9.5 was investigated using Sprague- Dawley rats. Irradiated rats and appropriate controls were killed at prenatal intervals of 4h, 48h and 10 days after exposure. Fetuses were examined for abnormalities and random samples of tissues were prepared for microscopic study. With the exception of the neuroepithelium, no histopathological changes were observed in embryos 4h after exposure to 50 cGy. In irradiated embryos, mitoses were reduced within the neuroepithelium; pyknosis and some necrosis of cells were apparent at this gestational interval. Among the gross developmental abnormalities observed in embryos 48h after irradiation, excessive flexion of the embryo and abnormal flexion of the head were the only ones that appeared to be radiation-induced. The mean numerical score (47.310.2, controls; 42.410.1, irradiated) for 17 morphological parameters examined in fetuses at this gestational period compares favorably with other studies. Controls, however, showed greater variability in the extent of development of their forebrain, olfactory system, midbrain, hindbrain, and caudal neural tube. In al1 cases, there was evidence of slower development in these regions compared to their irradiated counterparts. At term. no significant differences in litter size or resorption rates were observed in irradiated animals compared to the controls, but there was a higher incidence of defective eye development, spinal curvature and visceral anomalies. In utero exposure to 50 cGY gamma-radiation during the period of early organogenesis can produce some irreversible defects that are discernible at term.
Open Access
Heterogeneous immunoreactivity of frozen human benign and malignant breast lesions to C-MYC and C-Ha-ras cellular oncogenes
(Murcia : F. Hernández, 1994) pechoux, C.; Chardonnet, Y.; Chignol, M.C.; Noël, P.
C-myc and c-Ha-ras oncoprotein expression was studied by immunohistochemistry and gene detection by in situ hybridization on serial frozen sections of 32 breast lesions (19 benign biopsies and 13 infiltrating carcinomas). C-myc protein was expressed in 15/19 benign and 12/13 malignant lesions; c-myc gene was detected in 17/19 benign and 13/13 malignant lesions. Although a higher proportion of benign biopsies (819) showed more than 50% of protein-positive cells than malignant specimens, this cannot predict the outcome of a lesion. Conversely, p2 1 ras protein was expressed only in 2/19 benign lesions and in most cases of grade 1 to 111 carcinomas. The c-Ha-ras gene was always detected in a small percentage of cells, in both benign and malignant lesions. The results obtained with atypical hyperplasia, a doubtful proliferating lesion, suggests that p21 c-Ha-ras protein expression is not restricted to breast carcinomas. Although Southern blot is commonly considered as a very sensitive technique for oncogene analysis, no amplification of c-myc and c-Ha-ras gene has been demonstrated either in benign or malignant lesions. The detection, on serial frozen sections, of proteins and DNA of c-myc and c-Ha-ras, showed a possible amplification of the c-myc and c-Ha-ras genes in various benign and malignant lesions.
Open Access
Regional differences in cell surface patterns in normal human sulcular epithelium
(Murcia : F. Hernández, 1994) Sánchez-Quevedo, M. C.; Moreu, G.; Campos, Antonio; García, J. M.; González-Jaranay, M.
Studies with scanning electron microscopy in the normal human sulcular epithelium are scarce, and no precise information exists about cell surface patterns along the epithelium, the frequencies of these patterns, or possible regional differences within the mouth. In five periodontal biopsy specimens each from the anterior and posterior region of the mouth, we observed three cell patterns on the basis of the overall appearance of morphological surface markers in the coronal and apical zones of sulcular epithelium: microvilli; microplicae; and pits. The percentage of keratinocytes showing the microvillous pattern in the surface of apical sulcular epithelium of the posterior region of the mouth was significantly higher than in the anterior region. We posit that the presente, in the bottom of the normal sulcular epithelium in the posterior region of the mouth, of mainly microvillous keratinocytes (the most undifferentiated and least desquamative type of keratinocyte, and thus the most vulnerable to bacterial colonization) can be associated with observations of longitudinal clinical studies of periodontal disease, which suggest that more severe clinical findings are found in the region of the molars.
Open Access
A Golgi study on the red nucleus in man
(Murcia : F. Hernández, 1994) Patt, Stephan; Gerhard, Lieselotte; Zill, E.
The different cell types comprising the human red nucleus (RN) from eight patients without neuronal diseases were investigated using the Golgi- Braitenberg method for long-stored autopsy material. No giant cells were found due to regression of the magnicellular part of the human RN. We found larger (40 - 50 pm) and smaller (30 pm perikaryon size) medium-sized multipolar neurons with long dendrites, mushroom spines and typical dista1 dendritic tufts. The larger medium-sized RN neurons had some brushshaped dendritic end portions which could not be observed in the Golgi studies on various other mammals described in the literature. We additionally found small neurons with a perikaryon size of 15 pm. These cells were thought to be intrinsic neurons similar to those in animal investigations. The neuronal types found in the normal human RN corresponded to those in the parvicellular part of the mammalian RN. Dendritic end brushes, however. are typical only for the human RN.
Open Access
Histochemical and morphometric study of the cricoarytenoideus lateralis muscle in the horse
(Murcia : F. Hernández, 1994) López-Plana, C.; Sautet, J.Y.; Ruberte, J.; Sabaté, D.
Histochemical and morphometric parameters of the cricoarytenoideus lateralis muscle of the horse are presented. Using myosin ATPase staining after acid preincubation, 3 fibre types (1, IIA and IIC) were identified. Using NADH-TR staining, type 1 fibres showed high oxidative capacity, whereas type 11 fibres had high or low oxidative capacity. The type 1 to type 11 ratio was of 35:65. This ratio remained constant in the age range examined. Statistically significant (p<0.01) differences were found in values for fibre size between groups of horses weighing more than 500 kg and less than 400 kg. Mean area of type 11 fibres was greater (p<0.001) than that of type 1 fibres. There were no significant differences in mean area between left and right muscles in the group of animals with less weight. In contrast, significant differences (p<0.05) in mean area between left and right muscles were found for type 1 fibres in the group of animals exhibiting a higher weight. The histographical distribution of fibre type areas was unimodal. Most adult horses showed muscle fibre type grouping in the left muscle
Open Access
Histochemical study of the blue autofluorescence of collagen in oral irritation fibroma: Effects of age of patients and of the duration of lesions
(Murcia : F. Hernández, 1994) Dayan, D.; Wolman, M.; Hammel, I.
The intensity of the autofluorescence of collagen was measured in 27 irritation fibromata of the buccal mucosa and 13 of the lip. The intensity of fluorescence correlated positively with the duration of the lesion. The fluorescence intensity also increased with the patients' age. The present observations show that in irritation fibromata of buccal and lip mucosae, the intensity of blue autofluorescence of the collagen increases with duration of the lesions and with the age of patients.
Open Access
Ultrastructural and immunocytochemical study of the leptomeres in the mouse cardiac muscle fibre
(Murcia : F. Hernández, 1994) Hosokawa, T.; Okada, T.; Kobayahsi, T.; Hashimoto, K.; Seguchi, H.
The three-diiiiensional configuration and cheiriicol composition of leptoniei-es in the inouse \ e i i t r i c ~ i l a r cardiac inuscle fibre were electron iiiicroscopically und iminuiiocytochemically in\-estigated ~ising seinithin sections and specific antibodies agairist actin. the intei-mediate filament proteins desinin and viiiientin. anci the aciin-binding proteins a-actiriin, filainin and vinculin. The leptomeres appeared columnar in shape arid periodically segmeiited by electron-dense tiisc-like septa. The electrori-lucent areas betweeri these septa uere composed of fiiie interlinked filainents running obliquely to the major axis cif the Ieptomere. Actiii was localized in the electron-dense lii-ies of the leptoineres but not in the fine filaments. No reaction was. however. detected for desinin. vinientin. \zi~iculin. rilainin and a-actinin. The preserit results suggest. therefore. that the leptomeres inay not have a contractile function
Open Access
Ultrastructural alterations of the rabbit sciatic nerve, spinal cord and cerebellum, following methionine sulphoximine administration
(Murcia : F. Hernández, 1994) Kallaras, C.; Anogianakis, G.; Apostolakis, M.; Manthos, A.; Sioga, A.; Economou, L.; Foroglou, Ch.
Methii~nine suluhoximine (MSO) is a centrally ácting neurutoxin wh'ich inhibits the glutainate nietabolism enzymes and has convulsive properties. Sniall doses of MSO were administered to rabbits. either intra\,eiiously (IV) or intracerebroventricularly (ICV), anil electron microscopic examination of the cerebellum. the spinal cord and the sciatic nerve was perforined on the first day of rabbit hind leg rigid paralysis (myopathy with histvlogical findings resenibling myositis). which set in by [he 2nd to 4th day after MSO administration. In thc cerebelluin focal minor alterations were hund in the astrocytes (swelling and lucidity. diminution of glycogen graiiules) and sparsely in the presynaptic terminals (luciditj. and clumping). whereas inost of the neuron prescnted a nornial appearance. In the spinal cord and in the sciatic nerve a dissociation of the axon from the iiiyelin shcath was evident in a small number of niyelinatcd nerve fibres, along with the appearance of vacuolated spaces. Mitochondrial disorganisation in the axons. as well as glial cell alterations, were also seen. Th c u l t r a s t r u c t ~ ~ r aall terat ions we r e non spe c i f i c , Lind siiice they wcre induced 2 to 4 days after the adniiriistration of either minimum doses (IV) or of exti-cmely low doses (ICV) of MSO, they inay be attributed to the inordinate increase of metabolism during the period of convulsions.
Open Access
Effect of estrogenization in the first day of life on the reproductive system in male rats
(Murcia : F. Hernández, 1994) Limanowski, A.; Miskowiak, B.; Otulakowski, B.
The aim of the present report was to investigate dynamics of morphological and functional changes in the reproductive system of male rats between 20th and 84th day of life, injected neonatally with a single dose of stilbestrol. Marked reduction in relative weights of testes and accessory sexual glands was demonstrated in various periods of life. This was associated with inhibition of spermatogenesis at the stage of primary spermatocytes and with morphological as well as functional alterations in epididymis? seminal vesicles and ventral prostate. In the serunl, high levels of LH and lowered testosterone levels were demonstrated.
Open Access
Ultrastructure of spontaneous glomerular lesions in Syrian hamsters of APA strain
(Murcia : F. Hernández, 1994) Yamanouchi, J.; Yasoshima, A.; Han, J.S.; Itagaki, S.; Doi, K.
Electron microscopic observations were carried out on the spontaneous glomerular lesions in male APA hamsters from 3 to 12 months of age. Until 6 months of age, focal expansion of mesangial region due to an increase of matrix material and mesangial cells was characteristic, and segmental thickening of capillary basement membrane and partial effacement of foot processes of podocytes were also sometimes observed. At 12 months of age, although al1 of these changes became more severe, the most prominent alteration was found in podocytes, which showed various degenerative changes. No deposition of amyloid fibrils was detected in any portion of the glomerulus.
Open Access
Three types of liver cell dysplasia (LCD) in small cirrhotic nodules are distinguishable by karyometry and PCNA labelling, and their features resemble distinct grades of hepatocellular carcinoma
(Murcia : F. Hernández, 1994) Zhao, M.; zhang, N.X.; Du, Z.Y.; Laissue, J.A.; Zimmermann, Astrid
We have studied the occurrence and specific features of li\er cell dysplasia (LCD) in Chinese patients showing li\.er cirrhosis with or without hepatocellular carciiioina (HCC). Three types of LCD (SLCD, LLCDo. LLCDe) were rnorphologically defined. and these types were further analyzed using karyoinetry, estiniation of nucleic acid conteiit and deiisity, and PCNA iinmuiiostaiiiing. Features found for three types of LCD were coniparetl with those of normal hepatocytes (NLC). siinple regei~eratiiig hepatocytes (SRLC), and cells of HCCs covering different grades. The results show that 1 ) Aaryoinetry arid nucleic acid parameters allow an objecti\,e separation of LCD types both from NLC aiid SRLC; 2) karyometric features of LLCDe are most cloile to those of tiighly differentiated HCCs. whereas nuclear size nnd chroin~itinc omposition of SLCD closely 1-etlect those of poorly differentiated HCCs: 3) the frequency of LCD cluster.; was higher iii cirrhotic livers carrying HCC. being about douhle for al1 three LCD types: 4) the h i ~ h e sP~CN A labelling occurred in the small cell group of LCD (SLCD), still, however, beiiig smaller tlian that of simple i-egenerating hepatocytes. Based on these fiiidiiiys i t i \ suggested that, similar tu atypical adenoinatous hyperplasia, LCDs of distinct inorphotypes riiay represent precursor lesions for HCC, aiid soine cellular forms niay mimick cell types known to occur in experiinental carcinogenesis.
Open Access
Exoglycosidases and lectins as sequencing approaches of salivary gland oligosaccharides
(Murcia : F. Hernández, 1994) Menghi, Giovanna; Materazzi, G.
This review was focused on the salivary gland oligosaccharide chains studied by lectin histochemistry combined with exoglycosidase digestion. Glycoconjugates play an important role in many biofunctions and, generally. salivary mucins, which consist of numerous oligosaccharide chains attached at closely spaced intervals to a peptide backbone, serve as lubricants and protective agents, but in many instances we are ignorant about the role of biochemically identified oligosaccharides. Lectin histochemistry represents the greatest analytic tool to study carbohydrates in situ; in addition, there is availability of selective enzymes, so glycosidase degradation is useful to both investigate the structure of a given oligosaccharide and verify the influence of neighbouring sugars on the affinity towards the respective specific lectins. Using stepwise digestion of samples, followed by lectin labelling, the structure of terminal short oligosaccharides with blood-group activity was also elucidated. Additional histochemical methodologies were developed to establish the presence of acetylated groups in sialic acid residues, and the position of the linkage to the underlying monosaccharide. Sequencing approaches by exoglycosidases and lectins were also seen to be particularly useful when substantial differences did not emerge in lectin affinity, glycoconjugate composition and complex carbohydrate cytochemistry.