Histology and histopathology Vol.21, nº 9 (2006)
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- PublicationOpen AccessAdult stem and transit-amplifying cell location(Murcia : F. Hernández, 2006) Díaz-Flores Jr., L.; Gutiérrez, Ricardo; Varela, H.; Valladares, Francisco; Álvarez-Argüelles, H.; Díaz-Flores, Lucio; Madrid Cuevas, Juan FranciscoAdult stem cells (ASC) -able to self renew and to intervene in maintaining the structural and functional integrity of their original tissue- can express greater plasticity than traditionally attributed to them, adopting functional phenotypes and expression profiles of cells from other tissues. Therefore, they could be useful to regenerative medicine and tissue engineering. Transit-amplifying cells (TAC) are committed progenitors among the ASC and their terminally differentiated daughter cells. The ASC reside in a specialized physical location named niche, which constitutes a three-dimensional microenviroment where ASC and TAC are protected and controlled in their selfrenewing capacity and differentiation. The niche can be located near or far from the recruitment point, requiring a short or long-distance cellular migration, respectively. This paper briefly reviews the current status of research about ASC plasticity, transdifferentiation, fusion and functional adaptation mechanisms. Subsequently, ASC and TAC occurrence, characteristics and location have been considered in the skin, cornea, respiratory tract, teeth, gastrointestinal tract, liver, pancreas, salivary glands, kidney, breast, prostate, endometrium, mesenchyma, bone marrow, skeletal and cardiac muscle, nervous system and pituitary gland. Moreover, the role of cancer ASC has also been revised.
- PublicationOpen AccessElectron microscopic analysis of glucose-induced endothelial damage in primary culture: Possible mechanism and prevention(Murcia : F. Hernández, 2006) Mandal, A.K.; Ping, T.; Caldwell, S.; Bagnell, R.; Hiebert, L.M.We previously reported that high glucose treated cultured endothelial cells (ECs) showed intercellular gaps by transmission electron microscopy (TEM). These gaps were abrogated with insulin and/or heparin treatment. Our aims were to assess the severity of injury in ECs treated with high glucose for variable duration, and to further study the protective effects of insulin and/or heparin. Cells were also treated with Lbuthionine sulfoximine (BSO), a glutathione inhibitor, to help understand the mechanism of high glucose injury. Primary porcine ECs were treated with high glucose (30 mM) for 2, 6 or 10 days; and glucose plus insulin (1 U/ml), glucose plus heparin (5 µg/ml), glucose plus insulin plus heparin for 6 days. ECs were treated with BSO (0.001-0.05 mM) for 2 days. Pellets from trypsinized cells were processed for TEM. High glucose treatment revealed apoptosis or necrosis showing variable cell size, abnormal nuclei, condensation of nuclear chromatin, few mitochondria, cell membrane disruption and needle-shaped structures. Changes increased with duration of exposure. In high glucose plus heparin or insulin treated cultures at least one-half of the cells appeared normal. Most ECs were intact when treated with high glucose plus insulin plus heparin. BSO treatment showed dose-dependent changes with low doses showing apoptosis whereas higher doses revealed necrosis similar to high glucose treatment for 6 or 10 days. High glucose-induced EC injury increased with duration of exposure. These data demonstrate that high glucose injury resembles that of BSO treatment, suggesting that glutathione depletion may be involved in EC injury. Insulin and/or heparin protect against high glucose-induced injury.
- PublicationOpen AccessPlasma cell quantification in bone marrow by computer-assisted image analysis(Murcia : F. Hernández, 2006) Went, P.; Mayer, S.; Oberholzer, M.; Dirnhofer, S.Background: Minor and major criteria for the diagnosis of multiple meloma according to the definition of the WHO classification include different categories of the bone marrow plasma cell count: a shift from the 10- 30% group to the >30% group equals a shift from a minor to a major criterium, while the <10% group does not contribute to the diagnosis. Plasma cell fraction in the bone marrow is therefore critical for the classification and optimal clinical management of patients with plasma cell dyscrasias. The aim of this study was (i) to establish a digital image analysis system able to quantify bone marrow plasma cells and (ii) to evaluate two quantification techniques in bone marrow trephines i.e. computer-assisted digital image analysis and conventional light-microscopic evaluation. The results were compared regarding inter-observer variation of the obtained results. Material and methods: Eighty-seven patients, 28 with multiple myeloma, 29 with monoclonal gammopathy of undetermined significance, and 30 with reactive plasmocytosis were included in the study. Plasma cells in H&E- and CD138-stained slides were quantified by two investigators using light-microscopic estimation and computer-assisted digital analysis. The sets of results were correlated with rank correlation coefficients. Patients were categorized according to WHO criteria addressing the plasma cell content of the bone marrow (group 1: 0-10%, group 2: 11-30%, group 3: >30%), and the results compared by kappa statistics. Results: The degree of agreement in CD138-stained slides was higher for results obtained using the computer-assisted image analysis system compared to light microscopic evaluation (corr.coeff.=0.782), as was seen in the intra- (corr.coeff.=0.960) and inter-individual results correlations (corr.coeff.=0.899). Inter-observer agreement for categorized results (SM/PW: kappa 0.833) was in a high range. Conclusions: Computer-assisted image analysis demonstrated a higher reproducibility of bone marrow plasma cell quantification. This might be of critical importance for diagnosis, clinical management and prognostics when plasma cell numbers are low, which makes exact quantifications difficult.
- PublicationOpen AccessNeurofibroma with psammoma bodies(Murcia : F. Hernández, 2006) Kilmurray, L.G.; Ortega, L.; Martínez, A.; Sanz Esponera, J.Neurofibromas are benign tumours of the nerve sheath. Histologically they vary depending on their contents of cells, myxoid stroma and collagen. A 41-year old male with radicular pain had a tumour involving the posterior chest wall. Microscopically it resulted to be a neurofibroma with abundant psammoma bodies. Although these bodies are very frequent in some neoplasias, to our knowledge they have not been described in neurofibromas to date.
- PublicationOpen AccessMorphological effects of electrical stimulation and intermittent muscle stretch after immobilization in soleus muscle(Murcia : F. Hernández, 2006) Mattiello-Sverzut, A.C.; Carvalho, L.C.; Cornachione, A.; Nagashima, M.; Neder, L.; Shimano, A.C.The objective of the present study was to assess the effectiveness of a combined protocol of muscle stretching and strengthening after immobilization of the hindlimb. Thirty female Wistar rats were divided into 6 groups: group immobilized for 14 days to cause full plantar flexion by cast (GI, n=6); group immobilized/stretched (GIS, n=6): submitted to the same immobilization and to 10 days of passive stretching; group immobilized/electrically stimulated (GIES, n=6): similarly immobilized and submitted to 10 days of low frequency electrical stimulation (ES); group immobilized/stretched/electrically stimulated (GISES, n=6): similarly immobilized, submitted to 10 days of stretching and ES application; group immobilized/free (GIF, n=3): similarly immobilized and then left with free limbs for 10 days; control group (CG, n=3). The middle portion of the soleus muscle was frozen and sections were stained with HE or mATPase. Morphological analysis revealed high cellular reactivity in the GISES, GIES and GIS groups. The lesser diameter and proportion of type I fibers (TIF) and type II fibers (TIIF) (at pH 9.4) and connective area (at HE stain) were measured with an image analyzer and the data obtained were analyzed statistically by the unpaired Student t-test (p² 0.05). The results indicated that: a) immobilization generated atrophy of both fiber types (p<0.05); b) joint application of ES and stretching was not efficient in reestablishing the size of the two fiber types compared to CG (p<0.05); c) the ES protocol reestablished only the size of TIIF, which showed values similar to those detected in CG (p<0.05); d) the stretch increased the proliferation of the perimysium connective tissue (p>0,05). Thus, we conclude that, in the model applied here to female rats, a stretching protocol may limit the volume protein gain of soleus muscle fibers and increase the connective interstitial tissue.
- PublicationOpen AccessRegulation of EGFR endocytic trafficking by rab proteins(Murcia : F. Hernández, 2006) Ceresa, B.P.The Epidermal Growth Factor Receptor (EGFR) is a member of the receptor tyrosine kinase family and has important roles in development and cancer. Through ligand stimulation, the EGFR initiates a number of biochemical pathways that integrate to form specific physiological responses. In addition to these signaling pathways, the ligand stimulation also causes the EGFR to internalize and be transported through the endocytic pathway. The endocytic pathway regulates the rate of EGFR degradation and recycling, as well as the signaling mediated by the EGFR. In this review, the role of rabs, a family of small molecular weight guanine nucleotide binding proteins, is examined in how they regulate endocytic trafficking
- PublicationOpen AccessFhit protein is preferentially expressed in the nucleus of monocyte-derived cells and its possible biological significance(Murcia : F. Hernández, 2006) Zhao, P.; Hou, N.; Lu, Y.The FHIT gene encompassing the most active common human fragile region, FRA3B, has been proposed as a tumour suppressor gene for important common human carcinomas. The mechanism in which Fhit protein exerts its tumour suppressor activity is still obscure. To further understand the Fhit function associated with its intracellular localization we have investigated its cellular localization and distribution in human normal and cancerous tissues. Data of 1500 samples from immunohistochemistry showed that Fhit protein was preferentially and stably expressed in the nucleus of monocyte-derived or histiocytic lineage cells including monocytes of the circulating blood cells, macrophages of the connective tissue, Kupffer cells of the liver, alveolar macrophages or dust cells of the lung, osteoclasts of bone, microglia of the brain, epithelioid cells under chronic inflammatory conditions, foreignbody giant cells, Langerhans cells of the epidermis and dendritic cells of various kinds of human tissue, although the protein could also be infrequently observed in the nucleus of some quiescent epithelial cells. In active cells other than histiocytes, Fhit protein was detected either in cytoplasm or was negative. Neurons expressed Fhit strongly and neuroglial cells did so moderately but only in the cytoplasm. There was no Fhit protein detected in the neutrophils, lymphocytes, plasma cells and lipocytes. The present data showes that the stable nuclear localization of Fhit is not only a special marker for histiocytes with various morphologies but also may suggest the other function concerning Fhit as a signaling molecule related to anti-proliferation function. The detailed biological function related to nuclear localization of Fhit protein in the histiocytes remains to be further studied.
- PublicationOpen AccessCytoprotection by pyruvate through an anti-oxidative mechanism in cultured rat calvarial osteoblasts(Murcia : F. Hernández, 2006) Moriguchi, N.; Hinoi, E.; Tsuchihashi, Y.; Fujimori, S.; Iemata, M.; Takarada, T.; Yoneda, Y.Although we have previously shown drastic cell death by pyruvate deficiency in osteoblasts at the proliferative stage, the exact mechanism remains unclear so far. Cell survivability was significantly decreased in rat calvarial osteoblasts cultured for 0 to 3 days in vitro (DIV) following replacement of the eutrophic a- modified minimum essential medium (a-MEM) with Dulbecco’s modified eagle medium (DMEM) for cultivation. The addition of pyruvate enriched in a- MEM, but not in MEM, entirely prevented cell death induced by the medium replacement throughout a culture period from 0 to 3 DIV. Both cysteine and reduced glutathione protected cell death in cells cultured for 3 DIV without significantly affecting that in cells cultured for 1 DIV, however, while none of lactate, acetate and insulin significantly prevented the cell death irrespective of the culture period up to 3 DIV. A marked increase was detected in intracellular reactive oxygen species (ROS) levels 4 h after the medium replacement. In osteoblasts cultured in a-MEM for 3 DIV, but not in those for 7 DIV, hydrogen peroxide (H2O2) markedly decreased cell survivability when exposed for 2 to 24 h. Furthermore, H2O2 was effective in significantly decreasing cell survivability in osteoblasts cultured in DMEM for 7 DIV. Pyruvate at 1 mM not only prevented cell death by H2O2, but also suppressed the generation of intracellular ROS in osteoblasts exposed to H2O2. These results suggest that pyruvate could be cytoprotective through a mechanism associated with the anti-oxidative property rather than an energy fuel in cultured rat calvarial osteoblasts.
- PublicationOpen AccessStem cells, vascular smooth muscle cells and atherosclerosis(Murcia : F. Hernández, 2006) Margariti, A.; Zeng, L.; Xu, Q.Stem cells have the ability to differentiate into a variety of cells to replace dead cells or to repair tissue. Recently, accumulating evidence indicates that mechanical forces, cytokines and other factors can influence stem cell differentiation into vascular smooth muscle cells (SMCs). In developmental process, SMCs originate from several sources, which show a great heterogenicity in different vessel walls. In adult vessels, SMCs display a less proliferative nature, but are altered in response to risk factors for atherosclerosis. Traditional view on SMC origins in atherosclerotic lesions is challenged by the recent findings that stem cells and smooth muscle progenitors contribute to the development of atherosclerotic lesions. Vascular progenitor cells circulating in human blood and the presence of adventitia in animals are recent discoveries, but the source of these cells is still unknown. The present review gives an update on the progress of stem cell and SMC research in atherosclerosis, and discusses possible mechanisms of stem/progenitor cell differentiation that contribute to the disease process.
- PublicationOpen AccessHistological changes of liver glycogen storage in mice (Mus musculus) caused by high-protein diets(Murcia : F. Hernández, 2006) Ulusoy, E.; Eren, B.High Protein diets (HP) have been popular for people who want to lose weight since the 1960s. Even though these diets do not harm healthy people in the short term, there is insufficient data to support their safe use and efficiency over a long period. Because of the fact that the proteins in these diets are mainly from animal sources, it induces a higher intake of total fat, saturated fat and cholesterol. It is proven that high protein diets cause both physical and pathological abnormalities in the body. However, there exist very few studies about the effects of high protein nutrition on liver glycogen storage. For this study 40 Swiss albino mice consisting of two groups were used. The first group was fed with 25% High Protein; the other was fed with standard meal. The two groups were fed with respect to their diets for 30 days. At the end of 15th, 20th, 25th and 30th days 5 from each group were killed with cervical dislocation. The livers were removed after perfusion then fixated. The routine paraffin pursuit was applied before cutting into 5 µm sections and staining with H-E, PAS and silver. There were major differences in weight loss between the first and the fifteenth days. Glycogen storage was significantly reduced in HP (15) stained with PAS. Hydropic degeneration and regenerative activity was observed in H-E and silver stained HP group. As a result for the high protein diet group, weight loss at the 15th day and a significant decrement in glycogen storage at the 30th day was observed.
- PublicationOpen AccessNeurotrophins, airway smooth muscle and the fetal breathing-like movements(Murcia : F. Hernández, 2006) Inanlou, M.R.; Baguma-Nibasheka, M.; Keating, M.M.; Kablar, B.Central nervous system and skeletal muscles secrete a group of polypeptide hormones called neurotrophins (NTs). More recent studies show that NTs and their receptors are also expressed in the lung, suggesting a role for NTs in lung development. To examine the role of NTs during normal and diseased lung organogenesis, we employed wild-type and amyogenic mouse embryos (designated as Myf5-/- :MyoD-/-). Amyogenic embryos completely lacked skeletal muscles and were not viable after birth due to the respiratory failure secondary to lung hypoplasia. To examine the importance of lung-secreted NTs during normal and hypoplastic lung organogenesis, immunohistochemistry was employed. Distribution of NTs and their receptors was indistinguishable between normal and hypoplastic lungs. To further examine the importance of non-lung-secreted NTs (e.g., from the skeletal muscle and CNS) in lung organogenesis, in utero injections of two NTs were performed. The exogenously introduced NTs (i.e., non-lung-secreted) did not appear to improve development of the lung in amyogenic embryos. Moreover, immunohistochemistry showed significantly reduced number of airway smooth muscle cells (ASMCs) in hypoplastic lungs of amyogenic embryos, suggesting that the number of ASMCs is primarily regulated by the fetal breathing-like movements (i.e., mechanical factors).