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 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 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 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.
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