Histology and histopathology Vol.23, nº7 (2008)

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Open Access
Heat shock proteins and survivin, Relationship and effects on proliferation index of retinoblastoma cells
(Murcia : F. Hernández, 2008) Jiang, L.-B.; Liu, X.-Q.; Li, B.; He, X.-J.; Jin, Y.L.; Li, L.-Q.; Gao, F.; Wang, N.-L.
Survivin and HSPs (heat shock proteins) are important anti-apoptotic proteins. However, limited research has been done regarding the collective effects of HSPs and survivin on the proliferative activities of RB cells. The purpose of this study was to narrow this gap by focusing on the expression of HSP70 and HSP90 and the interaction of these proteins with survivin. The proliferative activities of RB cells were analyzed by assessing the Ki-67 labeling index. Ki-67 recognizes a nuclear antigen expressed in all phases of the cell cycle except G(0) and early G(1), which makes it an excellent marker of cells in the proliferative phase. Immunohistochemical procedures were performed on retinal tissues from 43 RB patients who had undergone enucleation. Expression of HSP70, HSP90 and survivin was found in 65.12%, 86.05% and 62.79% of the cases respectively. No expression of any of these markers was found in normal retinal tissues. Expression of survivin was more frequent when HSP90 was detected than when HSP90 was not detected (P<0.05). The Ki-67 labeling index was higher in cases in which HSP90 or survivin was found than in cases in which neither protein was found (P<0.05). The Ki-67 labeling index was higher in cases positive for both HSP90 and survivin than in cases in which neither protein or only one protein was found (P<0.05). Expression of HSP70 neither correlated with that of survivin, nor had any significant effect on the Ki- 67 labeling index (P>0.05). Although expression of HSPs and survivin and the Ki-67 labeling index did not correlate with histopathologic typing of RB (P>0.05), our findings demonstrate that expression of HSP90 correlates with that of survivin in RB and the coexistence of survivin and HSP90 probably plays an important role in cellular proliferation in RB. Further work is indicated to clarify the role of these processes in progression of RB.
Open Access
Lectin histochemistry for in situ profiling of rat colon sialoglycoconjugates
(Murcia : F. Hernández, 2008) Accili, Daniela; Menghi, Giovanna; Gabrielli, M.G.
The growing interest in glycoconjugates expressed and released by the epithelium of the intestinal mucosa is tightly related to the multiple functional roles attributed to sialic acid and its derivatives. In the present work, biotin and HRP conjugated lectins were used to detect the sialylation pattern and to identify specific structural features of sialoderivatives in the rat colon. In particular, the occurrence and distribution of sialic acids linked a2,6 to D-Gal/D-GalNAc and a2,3 to D-Gal were directly demonstrated with SNA and MAL II binding, respectively. In addition, in order to by-pass the specificity problems of SNA and MAL II as histochemical reagents, as well as to look for additional and complementary information about acetylation degree and sites, we combined sialidase digestion, potassium hydroxide deacetylation, and differential periodate oxidation with PNA and DBA binding. The data showed the distribution and structure of sialic acidß- D-Gal(1-3)-D-GalNAc and sialic acid-D-GalNac sequences, which proved to be widely distributed as cellular components or secretory products in surface goblet cells and crypt cells of the colonic epithelium. A high degree of O-acetylation, with acetyl groups mainly at 9 and 4 positions, was found, showing an increasing gradient from the proximal to distal portion of the colon. These results, which largely reproduce the sialylation pattern in other species, contribute new insights in defining the tissue specific expression of sialoderivatives in the colonic mucosa, and testify to their high heterogeneity which the wide range of sialic acid functional correlates in the intestinal tract depend on.
Open Access
Expression of inwardly rectifying K+ channels in the carotid body of rat
(Murcia : F. Hernández, 2008) Yamamoto, Y.; Ishikawa, R.; Omoe, K.; Taniguchi, K.
The inwardly rectifying K+ channels, Kir1.1, Kir2.3, Kir4.1-Kir5.1, and Kir4.2-Kir5.1, are candidate chemosensory molecules for CO2/H+. Here, we determined the mRNA expression and immunohistochemical localization of these channels in the carotid body (CB) and petrosal ganglion (PG) of the rat. RT-PCR analysis revealed mRNA expression of Kir4.1 and Kir5.1 in CB, and Kir1.1, Kir4.1, and Kir5.1 in PG. Immunohistochemistry identified the glomus cells in CB to express both Kir4.1 and Kir5.1 protein, while the nerve fibers in CB were immunoreactive for Kir1.1, Kir4.1, and Kir5.1. In the PG, immunoreactivity for Kir1.1, Kir4.1, and Kir5.1 was observed in some ganglion cells. Our findings suggest that Kir channels in the peripheral chemoreceptors play a role in sensing hypercapnic acidosis and maintaining the resting membrane potentials.
Open Access
Expression of the Shwachman- Bodian-Diamond syndrome -SBDS- protein in human pancreatic cancer and chronic pancreatitis
(Murcia : F. Hernández, 2008) Kayed, Hany; Bekasi, Sandor; Keleg, Sehereen; Welsch, Thilo; Esposito, Irene; Shimamura, Akiko; Michalski, Christoph W.; Friess, Helmut; Kleeff, Jörg
Background: The Shwachman-Bodian- Diamond syndrome (SBDS) protein is a member of a highly conserved family which influences RNA activation and is associated with pancreatic, skeletal and bone marrow deficiencies, as well as hematological malignancies. Methods: In this study, the expression and localization of SBDS were investigated in normal human pancreatic tissues, chronic pancreatitis (CP) tissues, primary and metastatic pancreatic ductal adenocarcinoma (PDAC) tissues, as well as in cultured pancreatic cancer cell lines by immunohistochemistry, immunoblotting and immunocytochemistry. Results: In the normal pancreas, SBDS was localized in the cytoplasm of islet cells and ductal cells. In CP tissues, SBDS was found in the cytoplasm of ductal cells, tubular complexes, stromal fibroblasts and in PanIN1-2 lesions. In PDAC tissues, SBDS exhibited cytoplasmic and occasionally nuclear localization in tubular complexes, PanIN1-3 lesions, cancer cells, and stromal fibroblasts. Different levels of SBDS protein were detected in cultured pancreatic cancer cell lines. Conclusion: SBDS is expressed in normal, CP, and PDAC tissues, as well as in pancreatic cancer cell lines. The different expression and localization patterns suggest a role of SBDS in the pathogenesis of, or response to, inflammatory and neoplastic pancreatic diseases.
Open Access
Involvement of FGF and BMP family proteins and VEGF in early human kidney development
(Murcia : F. Hernández, 2008) Carev, Dominko; Saraga, Marijan; Saraga-Babic, Mirna
The spatial and temporal pattern of the appearance of the fibroblast growth factor proteins (FGF-8 and FGF-10), the bone morphogenetic proteins (BMP-2/4 subfamily and BMP-7) and the vascular endothelial growth factor protein (VEGF) was investigated in the human mesonephros and metanephros of the 5-9 week-old conceptuses. In the mesonephros, both FGF’s and BMP’s were found in all structures and their expression slightly decreased in the early fetal period. VEGF positivity appeared in all mesonephric structures, and increased in the fetal period coincidently with formation of the mesonephric blood vessel network. In the metanephros, FGF-8 first appeared only in the metanephric mesenchyme, but from the 7th week on, its reactivity increased and spread to other metanephric structures. FGF-10 positive cells appeared in all metanephric structures already in the 5th week, and slightly intensified with progression of development. Cell survival and nephrogenesis in the permanent kidney might be associated with the appearance of both growth factors. Both BMP-2/4 and BMP-7 displayed a similar pattern of reactivity in all metanephric structures, and their reactivity intensified with advancing development. Alterations in their pattern of appearance might lead to the formation of small and dysplastic kidneys. Already in the earliest developmental stages, VEGF protein appeared in all metanephric structures. At later stages, VEGF showed more intense reaction in the collecting system than in the differentiating nephrons and interstitium. Due to VEGF involvement in vasculogenesis and angiogenesis, abnormal VEGF appearance might lead to impaired formation of the blood vessel network in the human permanent kidney.
Open Access
Dendritic cell migration and lymphocyte homing imprinting
(Murcia : F. Hernández, 2008) Villablanca, Eduardo J.; Russo, Vicenzo; Rodrigo Mora, J.
For an effective adaptive immune response to occur, dendritic cells (DC), which are the most efficient antigen-presenting cells, must be able to sample the peripheral microenvironment and migrate towards secondary lymphoid organs (SLO) where they activate naïve lymphocytes. Upon activation, lymphocytes proliferate and acquire the capacity to migrate to extralymphoid compartments. Although the molecular mechanisms controlling lymphocyte homing to lymphoid and to some extralymphoid tissues have been described in significant detail, it is much less clear how DC migration is controlled. Do DC obey similar adhesion cues that lymphocytes do, or do they have their own “zip codes”? This is relevant from a therapeutic standpoint because effective DC-based vaccines should be able to reach the appropriate tissues in order to generate protective immune responses. Here, we discuss some of the mechanisms used by DC to reach their target tissues. Once DC arrive at their destination, they are exposed to the tissue microenvironment, which likely modulates their functional properties in a tissue-specific fashion. This local DC “education” is probably responsible among other things; for the acquisition of tissue-specific homing imprinting capacity by which DC instruct lymphocytes to migrate to specific tissues. Finally, we discuss how dysregulation of these signals may play a key role in disease.
Open Access
The cellular and subcellular localization of zinc transporter 7 in the mouse spinal cord
(Murcia : F. Hernández, 2008) Chi, Zhi-Hong; Ren, Hao; Wang, Xin; Rong, Ming; Huang, Liping; Wang, Zhan-You
The present work addresses the cellular and subcellular localization of the zinc transporter 7 (ZNT7, SLC30a7) protein and the distribution of zinc ions (Zn2+) in the mouse spinal cord. Our results indicated that the ZNT7 immunoreactive neurons were widely distributed in the Rexed’s laminae of the gray matter in all spinal segments examined. The ependyma cells of the central canal and glia cells in the white matter were also shown ZNT7-positive. The ZNT7 immunoreactivity was mainly detected in the perinuclear regions of ZNT7- positive cells in the spinal gray matter. For ependyma cells, the immunoreactivity of ZNT7 was detected in the cytoplasm near the lumina of the central canal. Ultrastructural localization showed that ZNT7 was predominately present in the membrane of the Golgi stacks. The double immunofluorescence studies confirmed this result. Other intracellular organelles including the endoplasmic reticulum, mitochondria and lysosomes were devoid of ZNT7-immunostaining. The chelatable Zn2+ ions in the spinal cord were found predominantly in the terminals of the neuron rather than the cell body in the gray matter. However, overlapping distribution of chelatable Zn2+ ions and ZNT7 was found in the ependyma cells. The present study supports the notion that ZNT7 may function to supply zinc ions to the newly synthesized metalloproteins in the secretory pathway of the spinal neuron and the ependyma cell.
Open Access
Fibroblast remodeling of adsorbed collagen type IV is altered in contact with cancer cells
(Murcia : F. Hernández, 2008) Maneva-Radicheva, L.; Ebert, U.; Dimoudis, N.; Altankov, G.
A series of co-culture experiments between fibroblasts and H-460 human lung carcinoma cells were performed to learn more about the fate of adsorbed type IV collagen (Coll IV). Fibroblasts were able to spatially rearrange Coll IV in a specific linear pattern, similar but not identical to the fibronectin (FN) fibrils. Coll IV partly co-aligns with fibroblast actin cytoskeleton and transiently co-localize with FN, as well as with ß1 and a2 integrin clusters, suggesting a cell-dependent process. We further found that this Coll IV reorganization is suppressed in contact with H460 cells. Zymography revealed strongly elevated MMP-2 activity in supernatants of co-cultures, but no activity when fibroblasts or cancer cells were cultured alone. Thus, we provide evidence that reorganization of substrate associated Coll IV is a useful morphological approach for in vitro studies on matrix remodeling activity during tumorigenesis.
Open Access
Cardiac ischemia and reperfusion in spontaneously diabetic rats with and without application of EGb 761, I. cardiomyocytes
(Murcia : F. Hernández, 2008) Schneider, Rick; Welt, Klaus; Aust, Wolfram; Löster, Heinz; Fitzl, Günther
Diabetic cardiomyopathy is known to result in increased mortality after ischemic events. Permanently increased oxidative stress with formation of oxygen-free radicals plays a key role in the development of specific heart muscle disease. Associated lesions include structural alterations to cardiomyocytes. Antioxidative treatment in addition to the usual insulin substitution would seem sensible in preventing or delaying long-term diabetic complications and protecting the myocardium against acute ischemic events. We investigated the effects of radical scavenger Ginkgo biloba extract EGb 761 against diabetes-induced damage to cardiomyocytes and additional ischemia/ reperfusion injury in spontaneously diabetic BioBreeding/Ottawa Karlsburg (BB/OK) rats, as a model of diabetic myocardium infarction. Morphological and morphometric parameters of heart muscles were analyzed by light and electron-microscopic techniques. We used immunohistochemistry to evaluate parameters of oxidative stress (superoxide dismutase [SOD]) and inducible nitric oxide synthase (iNOS) protein expression. Our results indicated that A) Diabetic myocardium appears more vulnerable to ischemia/ reperfusion damage concerning ultrastructure of cardiomyocytes (sarcomeres, vacuoles, mitochondria), expression of antioxidative enzymes (CuZnSOD, MnSOD), and iNOS than normal myocardium; B) Pretreatment of diabetic myocardium with EGb and additional ischemia/reperfusion leads to a relative improvement in myocardial ultrastructure compared to unprotected myocardium. In summary, EGb appears to be promising as an adjuvant therapeutic drug in diabetics with respect to ischemic myocardium injury. It may contribute to the prevention of late diabetic complications in diabetic cardiomyopathy.
Open Access
Islet dynamics, A glimpse at beta cell proliferation
(Murcia : F. Hernández, 2008) Yesil, Pinar; Lammert, Eckhard
Pancreatic islets consist of 60-80% beta cells, which secrete insulin, a hormone of profound importance in the regulation of carbohydrate, fat and protein metabolism. Beta cell death and/or dysfunction result in an insufficient amount of insulin that leads to high glucose levels in the blood, a metabolic disorder known as Diabetes mellitus. Many studies aiming to establish new therapeutic applications for this disorder are targeted at understanding and manipulating the mechanisms of beta cell proliferation and function. The present comprehensive review summarizes the advances in the field of beta cell renewal and focuses on three fundamental issues: (i) identification of the cellular origins of new beta cells in the adult, (ii) regulation of beta cell proliferation, and (iii) downstream signaling events controlling the cell cycle machinery. Although the source of new adult beta cells is still being debated, recent findings in mice show an important contribution of beta cell proliferation to adult beta cell mass. In conjunction with describing characterized beta cell mitogens and components of the beta cell cycle machinery, we discuss how manipulating the proliferative potential of beta cells could provide novel methods for expanding beta cell mass. Such an expansion could be achieved either through in vitro systems, where functional beta cells could be generated, propagated and further used for transplantation, or in vivo, through directed beta cell renewal from sources in the organism. Once established, these methods would have profound benefits for diabetic patients.
Open Access
WWOX tumor suppressor gene
(Murcia : F. Hernández, 2008) Yang, Jilong; Zhang, Wei
Loss of heterozygosity and chromosomal rearrangement of the WWOX gene, which is located at 16q23.3-24.1, have been detected in ovarian, breast, hepatocellular, and prostate carcinomas and in other neoplasias. This gene, which spans the common chromosomal fragile site 16D, contains 9 exons and encodes a 46 kDa WWOX protein that contains 414 amino acids. The evidence from cancer cell lines and primary tumor tissues suggests that WWOX is a tumor suppressor gene and that its inactivation contributes to cancer development. The results from studies of WWOX gene knockout cancer cells and a WWOX knockout mouse model partly confirm this hypothesis. The nature of the various proteins that the WWOX protein can interact with, such as c-Jun, TNF, p53, p73, AP- 2gamma, and E2F-1, suggests that WWOX plays a central role in tumor suppression through transcriptional repression and apoptosis, with its apoptotic function the more prominent of the two. However, there is not universal agreement that WWOX is a tumor suppressor gene. Further analysis is needed to reveal the true nature of WWOX.
Open Access
Prognostic significance of nuclear and cytoplasmic expression of metallothioneins as related to proliferative activity in squamous cell carcinomas of oral cavity
(Murcia : F. Hernández, 2008) Szelachowska, Jolanta; Dziegiel, Piotr; Jelen-Krzeszewska, Joanna; Jelen, Michal; Tarkowski, Radoslaw; Wlodarska, Iwona; Spytkowska, Barbara; Gisterek, Iwona; Matkowski, Rafal; Kornafel, Jan
Metallothioneins (MT) are low molecular weight proteins with high metal and cystein contents. This study was designed to test the hypothesis that cytoplasmic and nuclear MT expression are of prognostic importance in patients with squamous cell carcinomas of the oral cavity, treated by surgery with subsequent radiotherapy. The second aim of the study was to test the potential correlation between the nuclear and cytoplasmic MT expressions as compared to expression of proliferation markers and other clinicopathological variables. Material and Methods: The studies were performed on tumor samples from 50 patients with diagnosis of squamous cell carcinoma of the oral cavity floor or of oral part of the tongue. All the patients were subjected to radical surgery, accompanied by removal of lymph nodes and post-operative radiotherapy. Results: No significant correlation could be detected between percentage and intensity of MT expression on one hand and proportions of cells with Mcm-2 (minichromosome maintenance protein 2), Ki-67 expressions, nor the grade of malignancy (G) on the other. A significantly shorter survival was detected among patients with tumors of MT expression rated 9 or 12 according to the Remmele scale and among patients with a high percentage (> 50%) of nuclear MT staining. In mulivariate analyses, only OTT (Overall Treatment Time), lymph node involvement and high expression of Mcm-2 were found to be independent risk factors for decreased patient’s survival. Conclusion: This is relevant evidence that MT overexpression could be related to worse prognosis in patients with oral cancer. We have found no relationship between MT expression and proliferative activity.
Open Access
Immunolocalization of histamine H3 receptors on endocrine cells in the rat gastrointestinal tract
(Murcia : F. Hernández, 2008) Grandi, Daniela; Shenton, Fiona C.; Chazot, Paul L.; Morini, Giuseppina
The histamine H3 receptor (H3R) has been identified in the gastrointestinal tract of the rat by immunohistochemistry, using the first validated anti-H3 receptor antibody. Immunoreactivity to H3R was exclusively localized to the endocrine cells scattered in the gastrointestinal mucosa, with positive cells being prominently abundant in the gastric fundus, while they were rarely found in the other regions. In the fundus, positive cells were distributed in the lower half of the mucosa and their number significantly decreased after a 24 h-fasting period. Double-labeling studies were undertaken to identify the H3R-immunoreactive cell types in the fundic and antral mucosa. The H3Rimmunoreactive cells were positive for chromogranin A. In the fundus, approximately 90% of cells positive to H3R were also positive to the histamine-forming enzyme, histidine decarboxylase. None of the cells expressing H3R displayed immunoreactivity for gastrin, somatostatin or ghrelin. Location, the influence of food deprivation and colocalization with histidine decarboxylase indicate that H3R positive cells correspond to the enterochromaffin-like cells (ECL).
Open Access
Indices 7,8,9
(Murcia : F. Hernández, 2013-01-18)
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