Histology and histopathology Vol.26, nº2 (2011)
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- PublicationOpen AccessRegulated expression of galectin-3, a multifunctional glycan-binding protein, in haematopoietic and non-haematopoietic tissues(Murcia: F. Hernández, 2011) Sundblad, Victoria; Croci, Diego O.; Rabinovich, Gabriel A.Galectin-3 belongs to a family of highly conserved animal lectins characterized by their ability to recognize multiple N-acetyllactosamine sequences, which can be displayed on both N- and O-glycans on cell surface glycoconjugates. Although first identified in macrophages, galectin-3 (also called ‘Mac-2, εBP, CBP35 or L-29’) has been found to be widely distributed in several tissues and developmental stages where, depending on its extracellular or intracellular localization, it can display a broad diversity of biological functions including immunomodulation, host-pathogen interactions, embryogenesis, angiogenesis, cell migration, wound healing and apoptosis. In spite of the existence of several reviews describing the multifunctional properties of galectin-3, an integrated view of the regulated expression of this glycan-binding protein in different normal tissues is lacking. Here we attempt to summarize and integrate available information on galectin-3 distribution in normal haematopoietic and non-haematopoietic tissues, mainly in adulthood, with only a brief reference to its expression during embryonic stages. In addition, given the multiplicity of biological roles attributed to this protein, a brief description of galectin-3 functions is also included. Understanding how galectin-3 is regulated in normal tissues will contribute to a rational design of approaches aimed at modulating galectin-3 expression and subcellular localization for experimental and therapeutic purposes.
- PublicationOpen AccessSomatostatin 14 affects the pituitary-ovarian axis in infant rats(Murcia: F. Hernández, 2011) Nestorović, N.; Ristić, N.; Manojlović-Stojanoski, M.; Šošić-Jurjević, B.; Trifunović, S.; Savin, S.; Milošević, V.The effects of multiple somatostatin (SRIH- 14) treatment on the pituitary-ovarian axis were examined in infant rats. Female Wistar rats received subcutaneously two daily 20 µg/100g b.w. doses for five consecutive days (from 11 to 15 days of age). Changes in cell volume, volume density and number per unit area (mm2) of follicle-stimulating (FSH), luteinizing (LH) and somatotropic (GH) immunolabeled cells were evaluated by stereology and morphometry. Serum FSH and LH concentrations were determined by RIA. Ovaries were analyzed by simple point counting of follicles. SRIH-14 treatment significantly reduced FSH and LH cell volume, while their volume density and number per unit area were unaltered. Serum concentrations of FSH and LH were significantly reduced. Volume and volume density of GH cells was significantly decresed after SRIH-14 treatment, while their number per unit area was unaltered. In the ovary, SRIH-14 induced a significant increase in the percentage of primordial follicles followed by a significant decrease in percentage of primary follicles. The number of healthy and atretic preantral follicles was unchanged. It can be concluded that SRIH-14 treatment during the infantile period markedly inhibits pituitary FSH, LH and GH cells. In the ovary, SRIH-14 acts by inhibiting initial folliculogenesis without affecting atretic processes.
- PublicationOpen AccessMedial artery calcification of uremic patients: a histological, histochemical and ultrastructural study(Murcia: F. Hernández, 2011) Ballanti, P.; Silvestrini, G.; Pisanò, S.; De Paolis, P.; Di Giulio, S.; Mantella, D.; Lappelli, M.; Favarò, A.; Bonucci, E.; Coen, G.Recent findings suggest that vascular calcification (VC) is an active process similar to bone mineralization, the vascular smooth muscle cells (VSMCs) undergoing phenotypic differentiation into osteoblastic cells and synthesizing calcificationregulating proteins found in bone. This study has investigated the VC process of uremic patients, with a morphologic approach. Epigastric artery samples from 49 uremic, non-diabetic patients were taken during kidney transplantation. Sections from paraffin-embedded samples were stained with hematoxylin/eosin and von Kossa. CD68 was immunohistochemically detected, and sections from frozen samples were stained with Oil Red O. Deeply calcified samples were stained with Picrosirius Red, PAS, and Alcian blue. Specimens from one patient with moderate and one with severe VC were examined under the electron microscope. None of the samples had atherosclerosis. Calcifications were found in the media of 38 patients. In 23, dot-like calcifications were irregularly scattered near the adventitia (light VC); in 11, granular calcifications formed concentric rings near the adventitia (moderate-advanced VC); in 4, zones of consolidated calcifications were found (severe VC). These zones were poor in collagen, glycoproteins and proteoglycans. In cases with moderate or severe VC, VSCMs showed necrotic changes. Matrix vesicles could be recognized in the extracellular spaces. In cases with severe VC, uncalcified or partially calcified membranous bodies were found, together with Liesegang rings. Patches of fibrin were also found. These findings point to a mainly degenerative mechanism of VC, which proceeds from the outer portion of the media. An active mechanism, however, cannot be excluded. A unifying hypothesis is suggested.
- PublicationOpen AccessPhysiopathologic dynamics of vesicle traffic in astrocytes(Murcia: F. Hernández, 2011) Potokar, Maja; Stenovec, Matjaž; Kreft, Marko; Gabrijel, Mateja; Zorec, Robert. The view of how astrocytes, a type of glial cells, contribute to the functioning of the central nervous system (CNS) has changed greatly in the last decade. Although glial cells outnumber neurons in the mammalian brain, it was considered for over a century that they played a subservient role to neurons. This view changed. Functions thought to be exclusively present in neurons, i.e. excitability mediated release of chemical messengers, has also been demonstrated in astrocytes. In this process, following an increase in cytosolic calcium activity, membrane bound vesicles, storing chemical messengers (gliotransmitters), fuse with the plasma membrane, a process known as exocytosis, permitting the exit of vesicle cargo into the extracellular space. Vesicles are delivered to and are removed from the site of exocytosis by an amazingly complex set of processes that we have only started to learn about recently. In this paper we review vesicle traffic, which is subject to physiological regulation and may be changed under pathological conditions.
- PublicationOpen AccessTime-course expression of metallothioneins and tissue metals in chronic relapsing form of experimental autoimmune encephalomyelitis(Murcia: F. Hernández, 2011) Jakovac, Hrvoje; Grebic, Damir; Tota, Marin; Barac-Latas, Vesna; Mrakovčić-Šutić, Ines; Milin, Čedomila; Radosevic-Stasic, BiserkaTo elucidate the role of metallothioneins (MTs) in the pathomechanisms of autoimmune CNS disorders we estimated the expression of MTs I+II and the tissue concentrations of Zn2+ and Cu2+ in the brain, spinal cord (SC) and in the liver during the periods of attacks and remissions in chronic relapsing experimental autoimmune encephalomyelitis (CR-EAE). Disease was induced in the genetically susceptible Dark Agouti (DA) rats by subcutaneous injection of bovine brain homogenate in CFA. Control rats were treated with CFA. The data, obtained by clinical assessment, immunohistochemistry and inductivity coupled plasma spectrometry, have shown that during the first attack (on the 12th day) MTs I+II were markedly upregulated in subarachnoid regions and perivascular space on astrocytes, microglia and on spinal neurons. Simultaneously, the concentrations of zinc in the SC and zinc and copper in the liver have found to be increased. During the second attack (on the 22nd day) a new overexpression of MTs was found in the cerebellum, in sulcus hippocampi, in spinal neurons and particularly in hepatocytes around the central vein. Concomitantly, in the brain and SC the concentration of copper increased. The data point to a neuroprotective role of MTs and to an important regulatory role of essential metals and hepatic MTs in the pathogenesis of CR-EAE
- PublicationOpen AccessExpression of stem cell marker CD133 in fetal and adult human kidneys and pauci-immune crescentic glomerulonephritis(Murcia: F. Hernández, 2011) Kim, Kyungeun; Park, Bong-Hee; Ihm, Hyojin; Kim, Kyung Min; Jeong, Jinuk; Chang, Jai Won; Cho, Yong MeeBackground: Different cell types in the kidney, including those in tubules, glomeruli, and interstitium, have been proposed as candidate adult renal stem cells, raising controversy about the very existence of such cells. In this study, we sought to clarify the location and nature of adult renal stem cells and to address their reparative role in native kidney disease. Methods: The expression of the stem cell marker CD133 was analyzed in 31 normal fetal and adult human kidneys by immunohistochemical methods. Co expression of CD133 with Ki-67 and tubule specific markers was also examined. Seventeen cases of pauciimmune glomerulonephritis were evaluated for CD133 and Ki-67 expression, and this was correlated with the patients’ renal prognosis. Results: CD133 was expressed in S-shaped bodies of fetal kidneys and co-expressed with Ki-67. It was highly expressed in mature tubules of fetal and adult kidneys without Ki-67 co-expression. CD133+ cells were most abundant in the S3 segment of proximal tubules and co-expressed with the distal tubule marker, suggesting multipotency. Most tubular CD133+ cells in normal adult kidneys exhibited pathologic features of acute and chronic injury. In pauci-immune glomerulonephritis, tubular CD133 and Ki-67 coexpression tended to be higher in cases where renal function recovered. Conclusion: These results suggest that adult renal stem cells reside predominantly in the S3 segment of the proximal tubule, where they remain mitotically silent under normal conditions, but can be induced by cellular injury. These results also suggest a potential role for adult renal stem cells in recovery from native human kidney disease.
- PublicationOpen AccessFOXP1 molecular cytogenetics and protein expression analyses in primary cutaneous large B cell lymphoma, leg-type(Murcia: F. Hernández, 2011) Espinet, Blanca; García-Herrera, Adriana; Gallardo, Fernando; Baró, Cristina; Salgado, Rocio; Servitje, Octavio; Estrach, Teresa; Colomo, Lluis; Romagosa, Vicenç; Barranco, Carlos; Serrano, Sergi; Campo, Elías; Pujol, Ramón M.; Solé, FrancescFOXP1 protein is expressed in normal activated B cells and overexpressed in a subset of diffuse large B-cell lymphomas, including primary cutaneous large B-cell lymphomas (PCLBCL), leg type. High expression of FOXP1 has been associated to an unfavourable prognosis with independent survival significance. However, little is known regarding the mechanisms underlying the overexpression of FOXP1 in PCLBCL, leg type. Our aims were to analyze FOXP1 cytogenetic status and protein expression in a series of PCLBCL, leg type. Finally, we compared the observed results with those obtained in a group of patients with primary cutaneous follicle centre lymphoma (PCFCL). Fifteen patients with PCLBCL, leg type and nine patients with primary cutaneous follicle centre lymphoma (PCFCL) were included in the study. For each biopsy specimen, FOXP1 translocation and copy number changes were evaluated by fluorescence in situ hybridization (FISH) and protein expression by immunohistochemistry (IHC). Immunohistochemistry showed FOXP1 staining in 13 PCLBCL, leg type, whereas all PCFCL were negative. FISH analysis disclosed no translocations involving FOXP1 gene in any of the cases. However, FOXP1 gene gains (3 to 4 copies) were observed in 82% of samples of PCLBCL, leg type and in 37% of PCFCL. FOXP1 expression was independent from FOXP1 translocation. Our results confirm that overexpression of FOXP1 is present in a considerable proportion of PCLBCL, leg type and might indicate an unfavourable prognosis. Mechanisms not related to translocation seem to be responsible for this overexpression.
- PublicationOpen AccessN-cadherin expression is involved in malignant behavior of head and neck cancer in relation to epithelial-mesenchymal transition(Murcia: F. Hernández, 2011) Nguyen, Phuong Thao; Kudo, Yasusei; Yoshida, Maki; Kamata, Nobuyuki; Ogawa, Ikuko; Takata, TakashiThe loss of E-cadherin and the gain of Ncadherin expression are known as “cadherin switching”. Cadherin switching is a major hallmark of epithelialmesenchymal transition (EMT). EMT is a crucial process in cancer progression, providing cancer cells with the ability to escape from the primary focus, to invade stromal tissues and to migrate to distant regions. Although down-regulation of E-cadherin is well known in various cancers, there are a few studies on N-cadherin expression in cancer. Here, therefore, we investigated whether N-cadherin expression was associated with the progression of head and neck squamous cell carcinoma (HNSCC). First, we examined the expression of Ncadherin by immunohistochemistry and its correlation with clinico-pathological findings. High expression of N-cadherin was observed in 52 of 80 HNSCC cases and was significantly correlated with malignant behaviors. Next, we examined the correlation between N-cadherin and E-cadherin. Cadherin switching (high expression of N-cadherin and low expression of E-cadherin) was found in 30 of 80 HNSCC cases and was well correlated with histological differentiation, pattern of invasion and lymph node metastasis in HNSCC cases. Moreover, we examined the expression of N-cadherin and E-cadherin by RT-PCR in 16 HNSCC cell lines to confirm the immunohistochemical findings. N-cadherin expression was observed in 7 of 16 HNSCC cells, and cadherin switching was observed in 2 HNSCC cells. Interestingly, HNSCC cells with cadherin switching have EMT features. In conclusion, we suggest that i) N-cadherin may play an important role in malignant behaviors of HNSCC, and ii) cadherin switching might be considered as a discrete critical event in EMT and metastatic potential of HNSCC.
- PublicationOpen AccessRole of nitric oxide in the regulation of fibrogenic factors in experimental liver fibrosis in mice(Murcia: F. Hernández, 2011) Leung, Tung-Ming; Fung, Man-Lung; Liong, Emily C.; Lau, Thomas Y.H.; Nanji, Amin A.; Tipoe, George L.Previously, we have shown that an increased expression level of iNOS but a reduction in the expression of eNOS is associated with increased oxidative stress markers in CCl4-induced experimental liver fibrosis. The present study aimed to investigate the effect of L-arginine and 5-methylisothiourea hemisulfate (SMT) in the expression of profibrogenic factors in chronic liver injury. ICR mice were treated with CCl4 with or without treatment of L-arginine, an NO donor, or SMT, an iNOS inhibitor. The expression of matrix metalloptroteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), α-smooth muscle actin (α- SMA), tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2) were investigated by RTPCR. The activity of the MMP-2 and MMP-9 were measured by zymography. Our results showed that CCl4- treated mice showed significant up-regulation of expression of pro-fibrogenic factors, TNF-α and COX-2. Treatment with L-arginine or SMT showed a significant reduction in CCl4-induced expression of these profibrogenic factors, TNF-α and COX-2. In conclusion, both SMT and L-arginine effectively attenuated the progression of CCl4-induced liver fibrosis. SMT suppresses iNOS mediated NO production. However, Larginine augments NO production. The similar effect of the two drugs on liver fibrosis indicates that there may be two distinct pathways of NOS mediated fibrogenesis in chronic liver injury by iNOS and eNOS. Our results suggest that eNOS-mediated liver fibrogenesis may play a more important role than that of iNOS in chronic liver injury. Taken together, these results support the contention that NO plays an active role in the progression of liver fibrosis and hepatocellular damage.
- PublicationOpen AccessA morphometric study of the spatial patterns of TDP-43 immunoreactive neuronal inclusions in frontotemporal lobar degeneration (FTLD) with progranulin (GRN) mutation(Murcia: F. Hernández, 2011) Armstrong, Richard A.; Cairns, Nigel J.Mutations of the progranulin (GRN) gene are a major cause of familial frontotemporal lobar degeneration with transactive response (TAR) DNAbinding protein of 43kDa (TDP-43) proteinopathy (FTLD-TDP). We studied the spatial patterns of TDP-43 immunoreactive neuronal cytoplasmic inclusions (NCI) and neuronal intranuclear inclusions (NII) in histological sections of the frontal and temporal lobe in eight cases of FTLD-TDP with GRN mutation using morphometric methods and spatial pattern analysis. In neocortical regions, the NCI were clustered and the clusters were regularly distributed parallel to the pia mater; 58% of regions analysed exhibiting this pattern. The NII were present in regularly distributed clusters in 35% of regions but also randomly distributed in many areas. In neocortical regions, the sizes of the regular clusters of NCI and NII were 400-800 µm, approximating to the size of the modular columns of the cortico-cortical projections, in 31% and 36% of regions respectively. The NCI and NII also exhibited regularly spaced clustering in sectors CA1/2 of the hippocampus and in the dentate gyrus. The clusters of NCI and NII were not spatially correlated. The data suggest degeneration of the cortico-cortical and cortico-hippocampal pathways in FTLD-TDP with GRN mutation, the NCI and NII affecting different clusters of neurons.
- PublicationOpen AccessQuantitative mRNA expression analysis in kidney glomeruli using microdissection techniques(Murcia: F. Hernández, 2011) De Spiegelaere, Ward; Cornillie, Pieter; Van Poucke, Mario; Peelman, Luc; Burvenich, Christian; Van den Broeck, WimThe introduction of new tools for molecular analysis, such as RT-qPCR and microarrays, has provided researchers with powerful applications to study renal disease and development. However, the high cellular heterogeneity of the renal tissue complicates the molecular analysis of specific cells and cell groups such as glomerular or tubular cells. In the past, glomerular sieving and manual dissection were used for the isolation of glomeruli. However, these techniques cannot be used for the isolation of specific glomeruli or for the coisolation of additional tissue fractions. In recent decades, new microdissection techniques such as laser-assisted microdissection have been developed. These applications allow the isolation of small cell groups from heterogeneous tissue for molecular analysis, including microarray and RT-qPCR. Although very promising, some drawbacks are associated with these techniques. The isolated sample material is generally small and requires sensitive assays. In addition, the long sample processing time may result in a considerable loss of RNA integrity. Careful optimization and rigorous quality analysis should overcome these drawbacks. In the present paper, the recent literature on the application of microdissection techniques in kidney research is reviewed, together with a discussion of the critical issues that are essential for the application of quantitative mRNA expression analysis with RT-qPCR on microdissected samples.
- PublicationOpen AccessHuman axillary apocrine glands: proteins involved in the apocrine secretory mechanism(Murcia: F. Hernández, 2011) Stoeckelhuber, Mechthild; Schubert, Christoph; Kesting, Marco R.; Loeffelbein, Denys J.; Nieberler, Markus; Koehler, Claudia; Welsch, UlrichThe apocrine secretory mechanism is a mode of secretion by which the apical part of the cell cytoplasm is pinched off, which leads to the formation of an aposome. The distinct mechanism of formation and decapitation of the aposome is not well investigated. Only few proteins are known that are involved in this secretory mechanism. We studied the human axillary apocrine gland and looked at proteins associated with cytokinesis, a process that is comparable to the pinchingoff mechanism of apocrine glandular cells. By immunohistochemistry, we detected actin, myosin II, cytokeratin 7 and 19, α- and ß-tubulin, anillin, cofilin, syntaxin 2, vamp8/endobrevin and septin 2. In highly active glandular cells, these proteins are located at the base of the apical protrusion when the aposome is in the process of being released or are concentrated in the cap of the apical protrusion. These findings demonstrate new insights on apocrine secretory mechanisms and point to similarites to the terminal step of cytokinesis, which is regulated by a SNARE-mediated membrane fusion event.
- PublicationOpen AccessComparative histopathological characteristics of highly pathogenic avian influenza (HPAI) in chickens and domestic ducks in 2008 Korea(Murcia: F. Hernández, 2011) Woo, G.H.; Kim, H.Y; Bae, Y.C.; Jean, Y.H.; Bak, E.J.; Kim, M.J.; Hwang, E.K.; Joo, Y.S.We compared characteristic lesions occurring in chickens and domestic ducks naturally infected with H5N1 HPAI virus in April and May 2008. Infected chickens generally exhibited pale-green, watery diarrhoea, depression, neurological signs and cyanosis of wattles and combs, and infected ducks generally exhibited neurological signs and watery diarrhoea. Gross petechial or ecchymotic haemorrhage affected the heart, proventriculus, liver, muscle, fat, and pancreas in chickens, and muscle in ducks. Necrotic foci were primarily present in the pancreas of both species and in the heart of domestic ducks. Histopathologically, chickens exhibited multifocal encephalomalacia, multifocal lymphohistiocytic myocarditis, multifocal necrotic pancreatitis and haemorrhage of several organs and tissues; ducks exhibited lymphohistiocytic meningoencephalitis with multifocal haemorrhages, multifocal necrotic pancreatitis, and severe necrotic myocarditis with mineralisation. The characteristic histopathologic findings of 2008 HPAI were multifocal encephalomalacia and necrotic pancreatitis accompanied by lymphohistiocytic myocarditis, and haemorrhage in various organs and tissues in chickens, whereas in ducks, they were severe necrotic myocarditis with mineralisation and necrotic pancreatitis, accompanied with lymphohistiocytic meningoencephalitis. The high mortality of domestic ducks may be intimately associated with heart failure resulting from increased H5N1 HPAI viral cardiotropism.