Histology and histopathology Vol.12, nº 4 (1997)

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Open Access
Alteration of cell cycle-related genes in hepatocarcinogenesis
(Murcia : F. Hernández, 1997) Nishida, N.; Fukuda, Y.; Ishizaki, K.; Nakao, K.
The mammalian cell cycle is controlled by regulators of the G1 to S transition such as tumor suppressor proteins, p53 and retinoblastoma (RB); cyclin D1 and cyclin-dependent kinase 4; and inhibitor of cyclin dependent kinase, Recently, aberrations of these cell cycle-related genes have been reported to contribute to the formation and development of cancer. In human hepatocellular carcinoma (HCC), high frequencies of aberration have been detected in the p53 and RB genes. Loss of heterozygosity (LOH) of chromosome 13q was detected in 35% of HCC and LOH on chromosome 17p was detected in 49%. Mutation of the p53 gene was also detected in 32%. The aberrations of these genes were observed more frequently in poorly differentiated and in advanced HCCs. On the other hand, genetic alterations of the cyclin DI and p161NK4A genes were not so frequent, but appeared to be associated with the aggressive behavior of the tumor, which suggests that disruption of the cell cycle-related genes results in the progression of HCC. Further study with a substantial number of cases is required to determine the actual frequency of the aberrations of the G1 controlling genes in hepatocarcinogenesis.
Open Access
Arteria1 microvascularization and breast cancer colonization in bone
(Murcia : F. Hernández, 1997) Yoneda, T.
Bone is one of the most preferential target organs of cancer metastases. Breast, prostate and lung cancers have a special predilection for colonization in bone. In an animal model in which inoculation of cancer cells into the left cardiac ventricle selectively develops osteolytic bone metastases but rarely forms metastases in non-bone organs, the pattern of breast cancer colonization in bone was studied radiologically and histologically. Colonization of cancer cells in bone was found to initiate and develop along with or at the terminal end of the major arteries running into bone. It should, therefore, be re-recognized that the anatomical vasculature still remains as a critica1 factor which influences cancer colonization in bone in addition to cellular and molecular properties of the bone microenvironment and metastatic cancer cells.
Open Access
Transcription factors in brain injury
(Murcia : F. Hernández, 1997) Pennypacker, K.
After brain injury, neuronal genes are regulated to adjust to an altered environment; however, if neurons are damaged then genes related apoptosis are activated. Glial cells, astrocytes and microglia, respond to neuronal death by transcribing genes to enhance the survival of remaining neurons and for regeneration and repair. AP-1 transcription factors are induced in the neuronal response to injury. Depending on the AP-1 dimer combination, neuronal genes related to either apoptosis or survival are transcribed. A 35 kDa Fosrelated antigen:JunD dimer is present in neurons that survive injury. Jun and JunD exists in neurons prior to undergoing apoptosis. Neuronal death activates gene expression in astrocytes and microglia. NFkB transcription factors are induced in astrocytes reacting to neuronal injury. In the microglial response, STATs appear to be activated to regulate gene transcription. These transcription factors that modulate the genes involved in the cellular processes of brain injury are examined in this review.
Open Access
Prohormone and proneuropeptide synthesis and secretion
(Murcia : F. Hernández, 1997) Perone, M.J.; Castro, M.G.
Hormones and neuropeptides in eukaryotic cells, are synthesised as large precursor molecules in the rough endoplasmic reticulum (RER), from where they are translocated to the Golgi apparatus. The sorting of proteins destined for the regulated secretory pathway from those which will be released constitutively takes place in the trans-Golgi network (TGN). In both these pathways, vesicles need to be transported to the plasma membrane before their contents can be released by exocytosis. Hormones and neuropeptides need to be secreted from the cells in which are synthesised to exert their biological actions, although they can also play paracrine and autocrine actions. Prohormones and proneuropeptides must undergo post-translational modifications which occur in determined subcellular compartments within eukaryotic cells and are carried out in a strict succession of intracellular events, which give rise to biologically active products. The biosynthesis of prohormones/proneuropeptides is mediated by the action of endoproteolytic enzymes and other post-translational modifying enzymes within the secretory pathway. The major focus of this review will be the biosynthetic pathway, sorting and intracellular trafficking of prohormone and proneuropeptide precursors within the secretory pathway of eukaryotic cells.
Open Access
Systemic mast cell disease (mastocytosis). General aspects and histopathological diagnosis
(Murcia : F. Hernández, 1997) Horny, H.P.; Ruck, P.; Krober, S.; Kaiserling, E.
Systemic mast cell disease/mastocytosis (SMCD) is best defined as a multitopic proliferation of cytologically andlor functionally abnormal tissue mast cells (TMC). SMCD preferentially involves the bone marrow, skin, spleen, liver, and lymph nodes. The histopathological diagnosis of SMCD may be very difficult to make, and the disease is often not considered in the differential diagnosis of lymphoreticular neoplasia. In suspected cases of SMCD, basic dyes such as Giemsa and toluidine blue are useful to demonstrate the specific metachromatic granules of TMC. The naphthol AS-D chloroacetate esterase reaction has also proved to be very reliable for enzyme-histochemical identification of TMC. Major diagnostic problems may arise in cases of malignant or «aggressive» SMCD exhibiting tissue infiltrates consisting predominantly of highly atypical, non-metachromatic TMC, which are usually also only weakly reactive for chloroacetate esterase. Immunostaining with antibodies against the mast cell-specific proteases tryptase and chymase has proved to be of great value for establishing the correct diagnosis in such cases. Anti-tryptase antibodies have major diagnostic significance due to their extremely high sensitivity and specificity. The classification of SMCD is controversial, but there is increasing support for the differentiation of at least two major subtypes that differ in prognosis: (i) a benign or «indolent» variant in which skin involvement (urticaria pigmentosa-like skin lesions) is usual, but associated malignant hematological disorders are rare; and (ii) a malignant or «aggressive» variant where skin involvement is usually absent but concomitant malignant hematological disorders (myelodysplastic and myeloproliferative syndromes and acute non-lymphocytic leukemias) are very common. Preliminary molecular biological studies of a few cases of malignant SMCD using the recently developed HUMARA assay have yielded evidence that the disease is monoclonal.
Open Access
Presence and distribution of 5HT-, VIP-, NPY-, and SP-immunoreactive structures in adult mouse lung
(Murcia : F. Hernández, 1997) Verastegui, C.; Fernandez-Vivero, J.; Prada, A.; Rodríguez, F.; Romero, A.
A large number of biologically active substances have been identified and characterised in the respiratory tract of severa1 mammals. These substances (amines and peptides) exert important regulatory influences on respiratory functions, and they act as neurotransmitters/neuromodulators, both being released from nerve terminals as neuroendocrine cells. However, these substances can also have other effects which suggest a paracrine action. Thus, to understand the role of amines and peptides in the lung, it is important to explore their localisation in different species. By using immunocytochemical staining methods we have studied the morphology and distribution of serotonin-, Substance P-, neuropeptide Y- and VIP-like immunoreactivity in the adult mouse lung. Moreover a pretreatment with colchicine, pargyline and 5-hydroxytryptophan as staining enlargement method was made. A widespread distribution of isolated endocrine cells and neuroepithelial bodies containing 5HT-like immunoreactivity was recorded within the lung. NPY-like immunoreactive nerve fibres were localised in the airway smooth muscle and surrounding the blood vessels. VIP-like immunoreactivity was revealed in single cells as well as in some nerve fibres and ganglia around the blood vessels and in the bronchial smooth muscle. SP-like IR was observed in nerve fibres located in the smooth muscle of the airways, surrounding bronchi and bronchioli but not next to the intrapulmonary blood vessels. Their localisation both in cells and nerve fibres of the respiratory system suggests that they play a role in the regulatory function of the mouse respiratory tract, exerting their influence by endocrine, paracrine, neurosecretory pathways or a combination of al1 of these.
Open Access
Cellular and molecular strategies for studying the regulation of bone resorption using the toothless (osteopetrotic) mutation in the rat
(Murcia : F. Hernández, 1997) Odgren, P.R.; Hermey, D.C.; Popoff, S.N.; Marks Jr., S.C.
The division of labor among cells of the skeleton is distinct and diverse and the regulation of these cells is interdependent. Osteoclasts are the cellular source of bone resorption and signals for their development and activation come, at least in part, from bone and other cells in the local environment. Studies of isolated cells have identified some factors in the developmental cascade of osteoclasts but there is little understanding of the sequence and local concentrations, not to mention other factors, needed for both the development of competent osteoclasts and for coordinated bone resorption. We review the skeletal biology of one osteopetrotic mutation in the rat, toothless, in which bone resorption is severely reduced because of a failure in the development and function of osteoclasts. Furthermore, we review the advantages and limitations of a relatively new method, differential display of mRNA (DD), that identifíes differences in gene expression in two or more populations of cells. We present a strategy and preliminary data for the application of DD to this mutation. We propose that application of this method to these and other skeletal diseases, with the appropriate controls and confirmations, will provide data about pathogenetic pathways and has a high probability for identifying new regulators of skeletal development and tumover.
Open Access
Stereological study of splenic tissue com partments in FK506-treated rats
(Murcia : F. Hernández, 1997) Milicevic, N.M.; Milicevic, Z.
Model-based stereology was used to study the changes of splenic structure and proportions of splenic tissue compartments in FK506-treated male Wistar rats (0.1 mg/kg three times a week for 14 days). The point-counting method was utilized to study the volume densities of the following tissue compartments: red pulp; white pulp (this compartment was divided in two subcompartments: follicles and periarteriolar lymphocyte sheath); marginal zone; aed connective tissue. The following stereological parameters of lymphoid follicles were also determined: areal numerical density (the number of follicles per mm2 of tissue section); the numerical density (number of follicles per mm3 of tissue); and the mean follicle diameter. After FK506 treatment a significant reduction of volume density of white pulp was observed, which was due to the very prominent decline in the amount of periarteriolar lymphocyte sheath, whereby the cellular density of this tissue compartment was decreased. In the marginal zone of FK506-treated spleens the presence of numerous, large, densely packed lymphoblastic cells of very uniform appearance was registered. These observations are in very good keeping with the changes of splenic morphology registered in cyclosporin Atreated rats, which further documents the parallelism in actions between these imrnunosuppressive agents.
Open Access
The effect of somatostatin analog octreotide on diethylstilbestrol-induced prolactin secretion, cell proliferation and vascular changes in the rat anterior pituitary gland
(Murcia : F. Hernández, 1997) Pawlikowski, M.; Kunert-Radek, J.; Grochal, M.; Zielinski, K.; Kulig, Andrzej
The effects of diethylstilbestrol (DES) and of long-acting somatostatin analog, octreotide (SMS) on the rat anterior pituitary microvasculature have been studied by means of computer-assisted image analysis. Additionally, the effects of DES and SMS on prolactin secretion and anterior pituitary cell proliferation have been studied, as well. The vascularization was visualized using Selye's method modified by Poely et al. (1964). The prolactin serum levels were estimated by radioimmunoassay. The proliferation indices were assessed using bromodeoxyuridine incorporation assay. As expected, it was found that DES sharply increased serum prolactin levels and enhanced cell proliferation in the anterior pituitary gland. DES also induced changes in parameters of vascularization. Simultaneous treatment of rats with SMS inhibited the DES-induced elevation of prolactin levels and pituitary cell proliferation. It also suppressed some but not al1 DES-induced changes in the anterior pituitary vascularization. These data suggest that the angio-inhibitory activity of SMS might be involved in its anti-tumor action on pituitary adenomas, but not as a sole or principal mechanism.
Open Access
Acidic glycosaminoglycans and laminin-1 in
(Murcia : F. Hernández, 1997) Center, Elizabeth M.; Emery, K.E.
Deposition of glycosaminoglycans and laminin-1 in the renal corpuscles of the kidneys of mylencephalic blebs, 'blebs', (my) and normal C57BLl 65 mice was compared in embryonic, neonatal (newbom to approximately two days old) and adult animals. Utilization of Alcian Blue 8GX staining, at a pH of 2.5, revealed an increase in acidic glycosaminoglycans in the parietal layer of Bowman's capsule and in general, an increase in the mesangial matrix of the glomerulus of my mutant adults. An increase in glycosaminoglycans was also noted in the developing kidney in certain my embryos in tissues associated with the glomeruli, but no significant differences were observed between the kidneys of neonatai my and control mice. The laminin-1 procedure revealed more deposition of laminin in the basement membrane of the parietal layer of Bowman's capsule in the neonatal and adult mutant my mice. Altered deposition of basement membrane and extracellular matrix components may reflect changes in the pattern of development and in the functioning of the kidney. Morphological changes in the human kidney are associated with alteration of function; a similar association may be occurring in the mice homozygous for the my gene.
Open Access
Silicone granulomatous lymphadenopathy and siliconomas of the breast
(Murcia : F. Hernández, 1997) Vaamonde, R.; Cabrera, J.M.; Vaamonde-Martín, R.J.; Jimena, I.; Marcos Martín, J.
In the present study, two histologicallydistinct cases of granulomatous lymphadenitis induced by dimethylpolysiloxane (silicone polymer) implants were studied. Four and six years after implant, and following surgery for breast cancer, painful homolateral axillary adenopathies were observed and biopsied. In both cases, histological examination led to a diagnosis of "silicone-induced granulomatous adenitis" requiring removal of implants. Foreign-body granulomas (siliconomas) were observed in surrounding tissue with no apparent rupture of implant capsules; however, visible retraction, hardening and scattered calcifications were noted. The presence of silica was revealed by incineration of a number of biopsied lymph nodes, a technique not hitherto used in the study of this pathology. A review is offered of the literature available
Open Access
lmmunocytochemical study on the innervation of the chicken pancreas by vasoactive intestinal polypeptide (VIP)-containing nerves
(Murcia : F. Hernández, 1997) Hiramatsu, K.; Oshima, K.
Ultrastructural localization of vasoactive intestinal polypeptide (VIP) was studied in the chicken pancreas by the immunocytochemical method. VIPimmunoreactive nerve endings were found in contact with acinar cells in the pericapillary space or between acinar cells. They also terminated on epithelial cells of arteriolae. In pancreatic islets, VIP-immunoreactive nerve endings were in contact with B-cells, but not with A- or D-cells. VIP-imrnunoreactive terminals contained many small clear vesicles which indicate the cholinergic feature. Nerve endings containing many small clear vesicles but not showing VIP-immunoreactivity were also detected in both the exocrine and endocrine pancreas. According to these data and the previous studies concerning the cholinergic and VIPergic innervation of the chicken pancreas, it is suggested that VIP-containing cholinergic nerves innervate B-cells and cholinergic nerves not containing VIP innemate D-cell.
Open Access
Bronchial epithelium associated to lymphoid tissue does not selectively express vimentin
(Murcia : F. Hernández, 1997) Alonso, L.M.; Cortés, A.; Barrutia, M.G.; Romo, T.; Varas, A.; Zapata, A.G.
The existence of a lymphoepithelium containing M cells in the bronchus-assmiated lymphoid tissue (BALT) of severa1 species has repeatedly been questioned. In previous electron microscopical studies we failed to ultrastmcturally identify these cells in the epithelium covering bronchial lymphoid tissue of adult rats. In the present study, we analyze immunohistochemically the expression of vimentin, an intermediate filament, reported to be a sensitive marker for rabbit M cells, in both BALT and Peyer's patches. Our results demonstrate, however, the absence of vimentin expression in the epithelium covering the bronchial lymphoid aggregates as well as in the lymphoepithelium of the Peyer's patches. On the contrary, both epithelia are strongly cytokeratin positive. Furthermore, numerous vimentin-positive lymphocytes appear in both lymphoid organs. Results are discussed from a view of the possible relationship between BALT and the so-called mucosaeassociated lymphoid tissue (MALT).
Open Access
Desmosomes and disease
(Murcia : F. Hernández, 1997) Chidgey, M.A.J.
Considerable progress has been made in our knowledge of desmosomes and their components. Molecular cloning of the desmosomal glycoproteins has established that desmoglein 1 and desmoglein 3 are targets for autoantibodies in the blistering diseases pemphigus foliaceus and pemphigus vulgaris respectively. New evidence suggests that another desmosomal glycoprotein, desmocollin 1, is the major target antigen in the upper epidermal form of intercellular IgA dermatosis (IgA pemphigus). In human cancer there is accumulating evidence which suggests a role for desmosomes in the prevention of invasion and metastasis. The possibility exists that a mutation in a desmosomal glycoprotein gene is responsible for an inheritable human disease, the striated form of palmoplantar keratodenna.
Open Access
When intracellular pathogens invade the frontiers of cell biology and immunology
(Murcia : F. Hernández, 1997) Pizarro-Cerdá, J.; Moreno, E.; Desjardins, M.; Gorvel, J.P.
Cellular microbiology has recently been described as a new discipline emerging at the interface between cell biology and rnicrobiology (Cossart et al., 1996). Many microbial pathogens can enter eukaryotic cells and live intracellularly either inside vacuoles or in the cytoplasm. The different steps during the invasion process are on the way of being dissected at the molecular leve1 revealing new insights in basic cellular functions. Indeed, bacterial pathogenesis can help us to better understand the dynamics of cell cytoskeleton, intracellular membrane traffic and signal transduction events. The recent advancements in the field of microbial pathogenesis are creating a new cross-taik between cell biologists, microbiologists and immuno-logists. In this review, the different strategies used by several pathogens are presented and the mechanisms elaborated by host cells from the immune system to eliminate the parasites discussed.
Open Access
The growth of non-pyramidal neurons in the primary motor cortex of man: a Golgi study
(Murcia : F. Hernández, 1997) Prinz, M.; Prinz, B.; Schulz, E.
In Golgi-Cox-impregnated motor cortex (regio precentralis) of six children (36 weeks of gestation to 13 month after birth) four types of interneurons of laminae 11-VI have been investigated qualitatively and quantitatively and finally, compared with interneurons of a child (3 month of age) with Down's Syndrome. Al1 cell measurements have been made by using a computerized digital morphometric system. In order to study the developmental stage of interneurons in our cases we statistically compared analogue neuron types in the brains. As pararneters for characterizing non-pyramidal neurons served the pattem of dendritic branching, dendritic length for al1 dendritic orders within single dendritic fields, and of the whole neuron. The quantitative morphological study of dendritic arborizations revealed that bitufted and large multipolar intemeurons had significantly more branching material in the postnatal brains than their prenatal counterparts. These differences were due to the increase of both length and number of branches during perinatal period. Such constant increment in dendritic span and branching provides larger receptive areas and that should improve the development of connections in functional intracortical columns. The quantitative parameter area of dendritic fields and number of branching points of the brain with Down's syndrome revealed in comparison with normal brain of the same age differences such as lower dendritic areas and a higher arnount of branching points
Open Access
Apoptosis in hepatocellular carcinomas with neuroendocrine differentiation
(Murcia : F. Hernández, 1997) Zhao, M.; Zimmermann, Astrid
We have studied apoptosis in a subset of thirty hepatocellular carcinomas (HCCs) exhibiting neuroendocrine differentiation (ND). Apoptosis was assessed by use of in situ DNA end labelling, and was quantified employing a TdT labelling index. It turned out that apoptosis occurred in HCCs with ND, albeit at different rates. Apoptosis was visualized as a clearly detectable reaction product mostly localized in tumor cell nuclei and in apoptotic bodies. Almost no staining was observed in nuclei of multinuclear tumor giant cells. In HCCs with ND, apoptosis was not related to type or grade, but tumors mainly consisting of hepatocyte-like cells andlor clear cells showed a significantly higher apoptotic rate. This was also the cell type most frequently disclosing neuroendocrine features. The apoptotic rate was significantly higher in HCCs with ND han in a control group of HCCs not showing ND. The findings suggest that neuroendocrine differentiation in liver cell tumors is associated with an aitered pattem of programmed cell death, and that this phenomenon may therefore have an influence on clinical behavior.
Open Access
Prolonged oval cell proliferation with Ito cell activation and extracellular matrix accumulation in galactosamine-induced acute hepatitis in mini rats
(Murcia : F. Hernández, 1997) Uetsuka, K.; Suzuki, M.; Nakayama, Hiroyuki; Doi, K.
Histopathological and immunohistochemical studies were carried out on D-galactosamine (Ga1NAc)- induced acute hepatitis in rats of the JCl: Wistar TgN (ARGHGEN) 1 Nts strain (Mini rats), in which expression of the growth hormone gene is suppressed by an antisense transgene. Hepatitis characterized by hepatocellular acidophilic necrosis with inflammatory cell infiltration was most prominent at 2 days after GalNAc (1000mglkg)-injection, when proliferation of Ito cells and deposition of fibronectin and laminin were found along the sinusoidal linings. At 72 hours after GalNAc-injection, Ito cell proliferation with deposition of laminin and fibronectin became more prominent, and marked proliferation of small epithelial cells was observed in the periportal area. At 7 days after GalNAcinjection, quite a number of a-smooth muscle actinpositive Ito cells, surrounded by abundant fibronectin, laminin and type IV collagen, were still observed in close juxtaposition to rapidly proliferating small epithelial cells. The small epithelial cells were found to be positive for both a-fetoprotein and cytokeratin 7 and were therefore considered to be so-called oval cells. The results suggest that there may be some relation between oval cell proliferation, Ito cell activation and extracellular matrix accumulation in GalNAc-induced acute hepatitis in Mini rats.
Open Access
Characterization of a monoclonal antibody recognizing mast cells
(Murcia : F. Hernández, 1997) Oliveira-Nogueira, T. de; Aubin, J.E.
Immunohistochemical screening for monoclonal antibodies prepared by immunization of mice with a rat osteoblastic cell population led to identification of one antibody that reacted against a small population of cells present in the soft connective tissue compartment of 21 days fetal rat calvaria. The morphology of the cells and the immunohistochemical staining characteristics (a distinct intracellular granular pattern) suggested that the antibody might be reacting specifically against mast cells. We used combined histochemistry and immunohistochemistry to further characterize this antibody, designated RCJ102. Cryosections containing calvaria bone, soft connective tissues and skin were prepared from the top of the head of 21 days fetal rats, and from adult rats cryosections of lung, muscle, adipose tissue and small intestine were prepared. Some sections were labelled by indirect immunofluorescence with RCJ102; corresponding sections were labelled histochemically with toluidine blue. There was a direct correspondence between mast cells identified histochemically and cells labelling with RCJ102 in al1 tissues except intestine, in which the mast cell detectable by histochemistry were not labelled by RCJ102. These results suggest that the RCJ102 antibody will be a valuable new reagent for further elucidation of the heterogeneity described between connective tissue and intestinal mucosal mast cells.
Open Access
Histopathology and clinical assessment correlate with the cysteine-serine-valinethreonine- cysteine-glycine (CSVTCG) receptor of thrombospondin-1 in breast tumors
(Murcia : F. Hernández, 1997) Roth, J.J.; Reiver, D.M.; Granick, M.S.; Rothman, V.L.; Nicosia, R.F.; Tuszynski, G.P.
Thrombospondin-1 (TSP-1) is a matrix protein implicated in mechanisms of tumor metastasis. TSP-1 has a characteristic Cysteine-Serine-Valine- Threonine-Cysteine-Glycine (CSVTCG) sequence that functions as a tumor cell adhesion domain. Our laboratory has isolated a novel CSVTCG specific tumor cell receptor. Immunohistochemical staining techniques and computerized image analysis were used to identify and quantitate the CSVTCG receptor of TSP-1 in a wide spectrum of human archiva1 breast tumors. Histopathologic and quantitative examination was correlated with clinical findings two years post operation. Increasing amounts of CSVTCG receptor correlated positively with worsening histopathologic and clinical findings. These findings suggest a role for the TSP-1 CSVTCG receptor in breast tumor progression. This receptor may have utility for the diagnosis, staging, and treatrnent of this common and deadly disease.