Histology and histopathology Vol.28, nº 6 (2013)
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- PublicationOpen AccessRoles of versican in cancer biology - tumorigenesis, progression and metastasis(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Du, William Weidong; Yang, Weining; Yee, Albert J.Versican, a large extracellular matrix proteoglycan accumulates in tumor stroma and plays a key role in both malignant transformation and tumor progression. Increased versican expression has been observed in a wide range of malignant tumors, and has been associated with both cancer relapse and poor patient outcomes in breast, prostate, and many other cancer types. Through negatively-charged chondroitin and dermatan sulfate side chains or interactions of the G1 and G3 domains, versican is able to regulate many cellular processes including cell adhesion, proliferation, apoptosis, migration, angiogenesis, invasion and metastasis. In this review, the biological roles that versican plays in cancer development are presented. Therapeutic targeting of versican in malignant tumors is also discussed.
- PublicationOpen AccessChanges in nitrosative stress biomarkers in swine intestine following dietary intervention with verbascoside(2013) Giancamillo, Alessia Di; Rossi, Raffaella; Vitari, Francesca; Carollo, Valentina; Deponti, Daniela; Corino, Carlo; Domeneghini, CinziaIn farm animals, oxidative stress can be involved in several intestinal pathological disorders, and many antioxidant molecules, especially those of plant origin, can counteract free radicals, thus stabilizing the gut environment and enhancing health. The aim of the study was to investigate whether the use of verbascoside (VB), a polyphenol plant compound, in pig feeding could modulate oxidative and/or nitrosative stress in the gut. Eighteen male piglets (Dalland) were assigned to two groups, which were fed with either a control diet (CON) or a diet supplemented with 5 mg/kg of verbascoside (VB) for 166 days. At slaughter, duodenum and jejunum specimens were collected. Immunohistochemistry and Western blot analyses were performed on the samples to evaluate free radical adducts, including acrolein (ACR), 8-hydroxydeoxyguanosine (8-OHdg) and nitrotyrosine (NT). A KRL test was also used to assess the total blood antioxidant activity, and no difference was observed. Immunohistochemistry and Western blot showed that dietary treatment decreased the levels of nitrotyrosine in enteroendocrine cell populations (P<0.05). Characterization of the enteroendocrine cell typology was then performed, and serotonin-immunoreactive cells were revealed to be directly involved in decreasing the nitrosative stress status. This preliminary study demonstrates the important role of dietary VB in decreasing stress biomarkers in swine gut, thus highlighting a possible intervention aimed at building a large prospective for antioxidant dietary supplementation in food animal species.
- PublicationOpen AccessInterleukin 1 receptor antagonist (IL1RN) genetic variations condition post-orthodontic external root resorption in endodontically-treated teeth(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Iglesias-Linares, Alejandro; Yañez-Vico, Rosa Mª; Ballesta-Mudarra, Sofía Ballesta-Mudarra; Ortiz-Ariza, Estefanía; Mendoza-Mendoza, Asunción; Perea-Pérez, Evelio; Moreno-Fernández, Ana Mª; Solano-Reina, Enriquey. External pical root resorption (EARR) is a frequent iatrogenic problem following orthodontic treatment in endodontically-treated teeth, about which the literature reports substantial variability in postorthodontic treatment EARR responses. The main focus of the present study is to clarify whether variants in the interleukin-1 receptor antagonist gene coding for the IL1ra protein have a positive/negative influence on EARR of endodontically-treated teeth. Ninety-three orthodontic patients were genetically screened for a single nucleotide polymorphism (SNP:rs419598) in the IL1 cluster. The sample was classified into 2 groups: group 1 (affectedgroup) showed radiographic EARR of more than 2mm; group 2 (control-group), had no EARR or EARR≤ to 2mm following orthodontic treatment on root-filled teeth. Logistic regression analysis was performed to obtain an adjusted estimate between the SNPs studied and EARR. Genotype distributions, allelic frequencies, adjusted odds ratios (OR) and 95% confidence intervals were also calculated. We found that subjects homozygous [1/1(TT)] for the IL1RN gene [OR:10.85; p=0.001;CI:95%] were at risk of EARR in root-filled teeth. Genetic variants in the antagonist axis balance of the IL1RN(rs419598) have a direct repercussion on the predisposition to post-orthodontic EARR in root-filled teeth. Variants in allele 1 of the interleukin-1 receptor antagonist gene(rs419598) are associated(p=0.001**) with an increased risk of suffering post-orthodontic EARR in root-filled teeth.
- PublicationOpen AccessImmunohistochemical analysis of angiotensin converting enzyme in sardinian pterygium(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Demurtas, Paolo; Di Girolamo, Nick; Corrias, Michela; Zucca, Ignazio; Maxia, Cristina; Diana, Andrea; Piras, Franca; Lai, Simone; Sirigu, Paola; Perra, TeresaPterygium is a common ocular surface disorder characterized by excessive cell proliferation, inflammation, fibrosis, angiogenesis and extracellular matrix remodeling. The Angiotensin converting enzyme (ACE or ACE I) is the major component of the Reninangiotensin system (RAS) converting the inactive decapeptide Angiotensin I (Ang I) to the active octapeptide Angiotensin II (Ang II). Besides this ‘classical role’, it can act as transcriptional regulator in response to external stimuli that may lead to cell damage and tissue remodeling. Due to this role, it can be internalized into the nuclear compartment to act as transcriptional factor for proteins involved in the inflammatory response. The aim of the present study was to determine ACE expression and localization in pterygium and culture pterygium cells by immunohistochemistry. Our results are the first to demonstrate nuclear immunolocalization of ACE, more so in pterygium compared to conjunctiva epithelial cells in histological sections. ACE was not detected in the nuclei of subcultivated pterygium epithelial cells. The nuclear localization of ACE may be correlated with an antiinflammatory path mediated by activation of its transcriptional role.
- PublicationOpen AccessNrf1 is time-dependently expressed and distributed in the distinct cell types after trauma to skeletal muscles in rats(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Zhang, Shu-Tao; Zhao, Rui; Ma, Wen-Xiang; Fan, Yan-Yan; Guan, Wen-Zheng; Wang, Jiao; Ren, Peng; Zhong, Kun; Yu, Tian-Shui; Pi, Jing-Bo; Guan, Da-WeiOur goal was to elucidate the dynamic expression and distribution of the nuclear factor erythroid-derived factor 2-related factor 1 (Nrf1) by immunohistochemistry, Western blotting, and real-time PCR during wound healing of contused skeletal muscle in rats. An animal model of skeletal muscle contusion was established in 40 Sprague-Dawley male healthy rats. Samples were taken at 6 h, 12 h, 1 day, 3 days, 5 days, 7 days, 10 days, and 14 days post-injury, respectively (5 rats in each posttraumatic interval). 5 rats were employed as control. A weak immunoreactivity of Nrf1 was observed in the sarcoplasm and nuclei of normal myofibers in control rats. Prominent immunostaining for Nrf1 was seen in a large number of polymorphonulcear cells, round-shaped mononuclear cells and spindleshaped fibroblastic cells, and regenerated multinucleated myotubes in the injured tissue. Subsequently, neutrophils, macrophages and myofibroblasts were identified as expressing Nrf1 by double immunofluorescent procedures. By real-time PCR analysis, Nrf1 expression was up-regulated and peaked at inflammatory phase. The expression tendency was also confirmed by Western blot. In conclusion, Nrf1 is time-dependently expressed in certain cell types, such as neutrophils, macrophages, myofibroblasts and regenerated multinucleated myotubes, suggesting that Nrf1 may modulate oxidative stress response and regeneration after trauma to skeletal muscles.
- PublicationOpen AccessDifferential expression of proteins related to smooth muscle cells and myofibroblasts in human thoracic aortic aneurysm(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Forte, Amalia; Della Corte, Alessandro; Grossi, Mario; Bancone, Ciro; Maiello, Ciro; Galderisi, Umberto; Cipollaro, MarilenaObjectives: Increasing knowledge is required for a better comprehension of the etiology of thoracic aortic aneurysm (TAA). The aim of this study was to highlight the modulations in vascular cell phenotypes, including myofibroblasts (MFs), in human TAA specimens compared to healthy aortas. Methods: histology, RT-PCR and immunohistochemical analysis of a panel of molecules, including EDA Fibronectin (Fn), smoothelin, CD34 and alpha-smooth muscle actin (alpha-SMA), selected on the basis of their informative potential as markers of smooth muscle cells (SMCs) and MF phenotypic modulation, were performed on all samples. Results: The media of TAAs was characterized by the absence of smoothelin, the unaltered expression of alpha-SMA accompanied by an alteration of its distribution pattern, and by the activated expression of the ED-A isoform of Fn. We found a concentration of round-shaped cells exclusively in the adventitia and in the perivascular tissue of TAAs, also rich in vasa vasorum, largely expressing alpha-SMA, while a sub-population also expressed ED-A Fn and CD34. CD34 was expressed by several cells in the intima of TAAs, together with cells expressing cytoplasmatic EDA Fn and alpha-SMA in comparison to healthy aortas. Conclusion: TAA specimens show an altered expression and localization of SMC and MF differentiation markers in comparison to healthy aortas, with possible implications on remodeling.
- PublicationOpen AccessImmunohistochemical expression of thymosin ß4 in ameloblastomas and odontomas(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Kiyoshima, Tamotsu; Nagata, Kengo; Wada, Hiroko; Fujiwara, Hiroaki; Shiotsuka, Maho; Kihara, Makiko; Hasegawa, Kana; Someya, Hirotaka; Sakai, HidetakaAmeloblastoma is regarded to be a benign odontogenic tumor, but it is destructive, locally invasive and presents a high rate of recurrence. Thymosin ß4 (Tß4) is closely associated with tooth germ development. Tß4 also plays a role in malignant progression and invasion. However, little is known about the function of Tß4 in odontogenic tumors. Thus, we investigated Tß4 expression in ameloblastomas and compared it with odontomas. We immunohistochemically evaluated the expression of Tß4, ameloblastin (AMBN), amelogenin (AMEL) and enamelin (ENAM) in 57 samples of ameloblastomas from 40 patients, and also assessed the expression of these molecules in 11 cases of odontomas, two of ameloblastic fibro-odontomas and one of tooth germ-like structures without the formation of enamel and dentin. Tß4 signals were observed in almost all of the ameloblastomas. The signals were observed in both peripheral columnar cells and central polyhedral/angular cells. Similar findings were observed in tooth germ-like structures, and in the ameloblastomatous nests in the ameloblastic fibro-odontomas. These samples had negative results for AMBN, AMEL and ENAM. Meanwhile, Tß4 signals were not seen in the odontomas, although immunolabeling for AMBN, AMEL and ENAM was observed in the enamel matrix and in some ameloblasts. Ectomesenhymal regions in the odontomas were negative for staining with the antibodies for AMBN, AMEL and ENAM. These results suggest that Tß4 could be associated with morphogenesis and tumor invasion in the ameloblastoma, and that Tß4 may play a role in the behavior of ameloblastoma.
- PublicationOpen AccessMast cell chymase: an indispensable instrument in the pathological symphony of idiopathic pulmonary fibrosis?(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Kosanovic, Djuro; Daha, Bhola Kumar; Wygrecka, Malgorzata; Reiss, Irwin; Günther, Andreas; Ghofrani, Ardeschir; Weissmann, , Norbert; Grimminger, Friedrich; Seeger, Werner; Schermuly, Ralph Theo; Banat, Gamal-AndreIdiopathic pulmonary fibrosis (IPF) is a chronic, progressive and fatal lung disease with no known etiology and treatment options. The hallmarks of the histopathology, which is characteristic of usual interstitial pneumonia (UIP) pattern, include interstitial fibrosis, honeycomb changes and fibroblast foci that develop owing to fibroblast proliferation and excessive matrix deposition. Although the complete pathomechanism is not yet understood, several molecular culprits, including transforming growth factor (TGF)-ß, Angiotensin (Ang) II, endothelin (ET)-1, matrix metalloproteinases (MMPs) and cytokines have been identified. IPF is increasingly believed to be an epithelial-driven disease; however, the literature does support an implication of altered immune response and inflammatory processes in the onset or progression of the disease. Mast cells (MCs) are multifunctional tissue resident cells involved in the inflammatory and immune response. An increasing body of evidence suggests a role of MCs and their mediator chymase in the pathology of IPF. With regard to the underlying mechanisms, it is conceivable that MC chymase may function via activation or processing of factors such as proteases, cytokines and growth factors. In this review, we will discuss how MC chymase is linked to and can potentially contribute to the development of IPF. Moreover, the findings from animal model studies will be discussed to highlight the chymase inhibitors as a promising strategy for the treatment of pulmonary fibrosis.
- PublicationOpen AccessReview of renal carcinoma with t(6;11)(p21;q12) with focus on clinical and pathobiological aspects(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Kuroda, Naoto; Tanaka, Azusa; Sasak, Naomi; Ishihara, AkiraHistology and Histopathology, vol. 28, nº 6, (2013); Matsuura, Keiko; Moriyama, Masatsugu; Nagashima, Yoji; Inoue, Keiji; Petersson, Fredrik; Martignoni, Guido; Michal, Michal; Hes, OndrejRecently, a new category of MiTF/TFE family translocation carcinomas of the kidney has been proposed. This category includes Xp11.2 renal cell carcinoma (RCC) and the t(6;11) RCC. These tumors share clinical, morphological, immunohistochemical and molecular genetic features. In this article, we review t(6;11) RCC. This tumor predominantly affects children and young adults. Macroscopically, the tumor generally forms a well circumscribed mass. Satellite nodules may be observed. Histologically, the tumor comprises large cells and small cells surrounded by basement membrane material. Immunohistochemically, tumor cells show nuclear immunolabeling for TFEB and usually express Cathepsin-K in the cytoplasm. Karyotyping detects the rearrangement between chromosome 6p21 and chromosome 11q12. Alpha-TFEB fusion can be detected by reverse transcriptase polymerase chain reaction (RTPCR) or fluorescence in situ hybridization (FISH). Most cases affecting children and young adults seem to be indolent, but some adult cases have presented with metastasis or caused death. As previously reported cases remain limited to date, further examination in a large scale study will be needed in order to elucidate clinical behavior and molecular characteristics.
- PublicationOpen AccessMechanism of experimental autoimmune neuritis in Lewis rats: the dual role of macrophages(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Shin, Taekyun; Ahn, Meejung; Matsumoto, Yoh; Moon, ChangjongHuman peripheral demyelinating diseases, such as Guillain-Barré syndrome (GBS), are characterized by inflammation and demyelination in the peripheral nervous system. Similarities in the pathology between GBS and the animal model of experimental autoimmune neuritis (EAN) indicate that autoimmune responses are involved in both diseases. This article summarizes the general aspects of the EAN model in Lewis rats and discusses the potential role of macrophages in the progression of EAN. A better understanding of macrophages may help to design alternative therapeutic strategies for organ-specific autoimmune diseases, including GBS.
- PublicationOpen AccessHistomorphometric and immunohistochemical study of the goat omasum during prenatal development(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) García, A.; Masot, Javier; Franco, Antonio; Gazquez, Antonio; Redondo, E.This work studies the morphological changes taking place in the goat omasum during prenatal development, using scanning electron microscope, light microscopy and immunohistochemical analysis. A total of 140 goat embryos and fetuses were used, from the first stages of prenatal life until birth. Differentiation of the omasum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life ([crown-rump length (CRL)] 3 cm, 23% gestation). By 38 days (CRL 4.3 cm, 25% gestation) the omasal wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Omasal laminae appeared in the following order: primary at 38 days (CRL 4.3 cm, 25% gestation), secondary at 50 days (CRL 7.7 cm, 33% gestation), tertiary at 59 days (CRL 12 cm, 39% gestation) and quaternary at 64 days (CRL 13.5 cm, 43% gestation). Neuroendocrine cells were detected by synaptophysin (SYP) at 52 days (CRL 8 cm, 35% gestation), while glial cell markers (glial fibrillary acidic protein - GFAP, and vimentin-VIM) were observed at 64 days (CRL 13.5 cm, 43% gestation) and 38 days (CRL 4.3 cm, 25% gestation), respectively. Sympathetic and parasympathetic nerve fibers and nerve bodies were detected via neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) at 95 days (CRL 20 cm, 63% gestation). In conclusion, prenatal development of the omasum - like that of the rumen - appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar stage to that reported in deer.
- PublicationOpen AccessExpression of integrins αvß3 and αvß5 and their ligands in primary and secondary central nervous system neoplasms(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Mittelbronn, Michel; Warth, Arne; Meyermann, Richard; Goodman, Simon; Weller, Michael. Aims: To study the expression of integrins αvß3 and αvß5 and their ligands in tumour, stroma and endothelial cells from human glioblastoma and CNS metastases from breast, lung and skin tumours. Methods and results: Integrin and integrin ligand expression was quantified in frozen tumour surgical specimens (15 glioblastomas and breast carcinoma metastases as well as 16 lung carcinoma and melanoma metastases) using immunohistochemistry. Gene expression profiles were evaluated in glioblastomas (n=424) and in normal brain (n=11). Overall, αvß3 expression was more common than αvß5, except in tumours derived from lung. αvß3 expression was most frequent in glioblastomas and melanoma metastases. Most lung-derived tumours expressed αvß5, but expression was less frequent in other tumours; about 20% of breast-derived tumours strongly expressed αvß5. Melanoma-derived tumours did not express αvß5. Expression of integrin ligands vitronectin, fibrinogen, fibronectin and osteopontin was variable between tumours, although most tumours expressed the ligands to some extent. Marked αvß3, but not αvß5, expression was common in stroma of CNS metastases. In blood vessels, αvß3 expression was more frequent than αvß5 and more pronounced in CNS metastases than in glioblastomas. Integrin ligand expression occurred in blood vessels in most tumours. In glioblastomas, mRNA expression of αvß3, αvß5, osteopontin and fibronectin were significantly upregulated over normal brain. Conclusions: Overall, we report distinct and heterogeneous patterns of integrin expression in primary and secondary brain tumours that may be relevant to the future development of integrintargeting therapeutic approaches to brain tumours.
- PublicationOpen AccessExpression of estrogen receptor, progesterone receptor, and Her-2/ neu in primary and extra-corporeal endometrial cancer(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Tangjitgamol, Siriwan; Tanvanich, Sujitra; Srijaipracharoen, Sunamchok; Manusirivithaya, SumonmalObjective: To compare immunohistochemical (IHC) expression of estrogen receptor (ER), progesterone receptor (PR), and Her-2/neu in the primary tumors of endometrial cancer (EMC) and their extra-corporeal lesions. Methods: Paraffin-embedded tissues of the primary and extra-corporeal tumors of EMC were retrieved for IHC study. Expression of ER, PR, and Her-2/ neu in the primary tumors and extra-corporeal lesions were compared. Results: From 72 EMC patients with 87 extracorporeal lesions, positive PR expression was significantly lower in the extra-corporeal lesions than that in the primary sites: 42.5% vs. 63.9%, respectively (p=0.007). No statistically significant differences of ER and Her-2/ neu expressions in the extra-corporeal and the primary sites were found: 42.5% and 55.6% for ER (p=0.102) and 20.7% vs. 13.9% for Her-2/ neu (p=0.262), respectively. The expression of extracorporeal lesions were concordant to the primary tumor in 65.5% of ER (k=0.319), 71.2% of PR (k=0.445), and 83.9% of Her-2/ neu (k=0.413). From 15 cases wherein IHC from two extra-corporeal sites were studied, 73% had concordant ER expression between the two extracorporeal lesions (k=0.412) while 93.3% had concordant PR and concordant Her-2/ neu expression (k=0.842 for PR and 0.634 for Her-2/ neu) Conclusion: PR expression was significantly higher in the primary tumors than the extra-corporeal sites. Higher ER and lower Her-2/ neu expressions in the primary tumors were also observed but the differences were not significant. The tumors heterogeneity suggests it may be important to study tumor tissues from both primary and extra-corporeal sites when planning treatment, especially by hormonal or targeted therapies.