Histology and histopathology Vol.31, nº8 (2016)
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- PublicationOpen AccessHistological and biochemical characteristics of the rabbit anterior cruciate ligament in comparison to potential autografts(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Hoyer, Mariann; Meier, Carola; Kohl, Benjamin; Lohan, Anke; Kokozidou, Maria; Schulze Tanzil, GundulaTissue engineering of an anterior cruciate ligament (ACL) implant with ACL cells requires detailed analysis of the tissue characteristics that should be mimicked. Therefore, we studied the histological and biochemical properties of rabbit derived ACLs in comparison to other knee-associated tendons that are used as autografts in men. Rabbit derived ACLs and Musculus (M.) semimembranosus, M. semitendinosus tendons and patellar ligaments were explanted from adult New Zealand white rabbits and analyzed histologically for tissue organization (e.g. cellularity, nuclear shapes, elastic fibers), total collagen and sulfated glycosamino-glycan (sGAG) contents. Gene expression analysis was performed for the main extracellular matrix (ECM) components type I collagen, decorin and the glycoprotein tenomodulin. The ACLs had an average dimension of 1.39x0.39x0.1 cm in situ. They were characterized by high sGAG content in comparison to the other tendons/ligaments, whereas the total collagen content did not differ. ACLs possessed higher cellularity and lower feret diameter of the cell nuclei compared with the investigated rabbit-derived tendons. In ACLs long elastic fibers were observed. Concerning the gene expression level, lower transcription of tenomodulin was detected in the ACL compared with the other tendons, without significant difference in the decorin gene expression. The M. semitendinosus tendon had a significantly higher type I collagen expression than the ACL and the other investigated tendons. This phenotypical characterization of the lapine ACL presented in this study provides some key standards to evaluate tissue engineered ACL constructs to be tested in the rabbit model.
- PublicationOpen AccessIodine deficiency induces a VEGF-dependent microvascular response in salivary glands and in the stomach(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Vanderstraeten, Jessica; Derradji, Hanane; Craps, Julie; Sonveaux, Pierre; Colin, Ides M.; Many, Marie Christine; Gérard, Anne CatherineDespite efforts to optimize iodine supply in iodine deficient countries, iodine deficiency (ID) remains a global problem worldwide. Activation of the local microvasculature by ID in the thyroid gland aims at improving the local supply of iodide. For this purpose, the thyrocytes secrete vascular endothelial growth factor (VEGF) that acts on adjacent capillaries, via a reactive oxygen species (ROS)/Hypoxia Inducible factor (HIF)- dependent pathway. Beside the thyroid, other organs including salivary glands and the stomach do express the sodium/iodide symporter (NIS) and are able to take iodide up, potentially rendering them sensitive to ID. To verify this hypothesis, ID-induced effects on the local microvasculature were studied in salivary glands and in the stomach. ID was induced by feeding young mice with an iodide-deficient diet and NIS inhibitor perchlorate in the drinking water. In salivary glands, ID induced a transient increase in HIF-1α protein expression accompanied by a transient, VEGFdependent increase in blood flow. In the gastric mucosa, ID transiently increased VEGF expression in the mucinsecreting epithelium and in ghrelin-secreting endocrine cells. These observations suggest that microvascular changes in response to ID occur in NIS-expressing tissues other than the thyroid. NIS expressing cells could be viewed as iodide sensors that respond to ID by inducing vascular changes, probably to optimize iodide bioavailability at regional or systemic levels.
- PublicationOpen AccessSteroid receptors in human ejaculated sperm as molecular markers of the detrimental effects related to the pathophysiology of testicular varicocele(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Perrotta, Ida; Aquila, SaveriaThe exact physiopathologic effect of testicular varicocele on male fertility is not defined yet. The detrimental role of the varicocele in fertility is supported by the presence of a higher frequency of affected men among the infertile population. However, the mechanism/s by which a varicocele impairs sperm production, structure and function, is not known. In spite of active interest, our understanding of the human male gamete ultrastructural molecular organisation is still incomplete and therefore our knowledge of the sperm molecular anatomy is very limited. The presence of steroid binding sites on human spermatozoa has been evidenced since the 1970s, and afterwards, spermatozoa physiology was linked to the action of different steroids. The presence of steroid/steroid receptor systems was demonstrated in mature spermatozoa as membrane but also as nuclear conventional receptors, suggesting that both systemic and local steroids, through sperm receptors, may influence male fertility. From new data, it emerges that varicocele may induce damage in the male gamete at molecular level, opening a new chapter in the already multifactorial pathophysiology of the varicocele, complicating this issue. In sperm from varicocele, a decreased expression of steroid receptors and a consequent reduced responsiveness to steroids may represent a mechanism involved in the physiopathology of varicocele. Therefore, the modulation of these nuclear receptors pave the way for novel therapeutic opportunities in the treatment of the male pathologies related to human reproduction. The purpose of this review is to gain new insight into the physiopathology of varicocele and to study its impact on human sperm molecular anatomy
- PublicationOpen AccessMolecular pathogenesis of hereditary hemochromatosis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Liu, Jingqi; Pu, Chunwen; Lang, Lang; Qiao, Liang; Abdullahi, Mohamud Abukar Haji; Jiang, Chunmeng. Hereditary hemochromatosis (HH) is an inherited iron overload disorder characterized by normal iron-driven erythropoiesis and abnormal iron metabolism, leading to excess iron deposited in parenchymal cells of liver, heart, and endocrine glands. Iron hormone, hepcidin, plays a critical role in iron homeostasis through interaction with ferroportin (FPN), a major cellular iron exporter. Hepcidin is encoded by hepcidin antimicrobial peptide (HAMP). Mutations in hepcidin and any genes that regulate the biology of hepcidin, including hemochromatosis genes (HFE), Hemojuvelin (HJV), transferring receptor 2 (TFR2) and FPN, result in hemochromatosis. The identification of hepcidin and its role will provide a better understanding for pathogenesis of HH.
- PublicationOpen AccessGastrospheres of human gastric mucosa cells: an in vitro model of stromal and epithelial stem cell niche reconstruction(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Santos, Carlos A.N.; Andrade, Leonardo R.; Costa, Márcia H.M.; Souza, Heitor S.P.; Granjeiro, José M.; Takiya, Christina M.; Borojevic, Radovan; Nasciutti, Luiz EuricoThe molecular characterization of mechanisms involved in the gastrointestinal tract disorders needs an in vitro 3D culture model able to mimic the in vivo gastric microenvironment. Herein, we propose a 3D coculture system where gastric epithelial and stromal cells are grown together building spherical and solid structures using the NASA bioreactor - cell culture system (RCCS), a bioreactor. Epithelial and stromal cells from human antral gastric mucosa were isolated from endoscopic gastric biopsies. Thereafter, these cells were mechanically and enzymatically dispersed by treatment with dispase and collagenase, respectively. Using specific culture procedures, these cells formed 3D structures by using a RCCS, named “gastrospheres”. Briefly, gastrospheres were obtained by initial seeding of 2.5x104 cells/well in 96 well culture plates. At 24 h after their formation, they were transferred into RCCS, and maintained for 7, 14, 21, and 28 days. The gastrospheres were morphologically characterized by immunocytochemisty to evaluate extracellular matrix (ECM), and by electron microscopy. These analysis of gastrospheres revealed that the epithelial cells were cytokeratin (CK) and lectin reactive and were arranged in the outer layer; stromal cells presented long cytoplasmic processes and were localized inside the gastrosphere. They were vimentin (VIM) and α-smooth muscle actin (α-SMA) positive and expressed ECM components such as laminin (LN), fibronectin (FN), and type IV collagen (CIV). Electron microscopy revealed groups of cohesive gastric cells surrounded by complex stromal structures, with multiple microvilli, and tight cellular junctions interspersed with extracellular matrix fibrils and fibers. The presence of some nestinpositive cells was observed in the inner region of the gastrospheres, suggesting an intermediary localization between epithelial and stromal cells. Altogether, our data suggest that in vitro gastrospheres recapitulate the in vivo gastric niche microenvironment.
- PublicationOpen AccessAberrant levels of Wnt/β-catenin pathway components in a rat model of endometriosis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Medina de Mattos, Rômulo; Rodrigues Pereira, Paula; Gouvêa de Oliveira Barros, Eliane; Henriques da Silva, Julianna; Yelimar Palmero, Celia; Meireles da Costa, Nathália; Ribeiro Pinto, Luis Felipe; Rodrigues Pereira Gimba, Etel; Hecht, Fábio; Bueno Ferreira, Luciana; Escorsim Machado, Daniel; Leite de Oliveira, Felipe; Eurico Nasciutti, LuizEndometriosis is a benign gynecological disease affecting approximately 10-15% of women of reproductive age and 25-50% of all infertile women. It is characterized by the presence of glands and/or endometrial stroma outside the uterine cavity. Angiogenesis is a crucial process for the development and maintenance of endometriotic lesions. The Wnt/βcatenin pathway is a major promoter of angiogenesis in both physiological and pathological conditions. In the present study, we evaluated the expression of molecules related to the Wnt/β-catenin pathway in a rat model of peritoneal endometriosis. mRNA analyses showed significantly increased expression of Wnt4 and Wnt7b and decreased expression of Gsk3beta and E-cadherin in endometriotic lesions. However, there were no differences in β-catenin and Fzd2 mRNA expression. In addition, we observed a significant increase of nuclear β-catenin in endometriotic lesions, a hallmark of Wnt/ β -catenin pathway activation. Stromal β-catenin staining was found in 45.4% of endometrial tissues and 77.8% of endometriotic lesions. β-catenin nuclear localization was found in 18.2% of the endometrial tissues and 33.3% of endometriotic lesions. Finally, the expression of galectin-3, a regulator of this pathway, was increased in endometriosis. In summary, this pattern of Wnt/βcatenin components expression suggests an increased activity of this pathway in endometriosis.
- PublicationOpen AccessAnalysis of the expression and localization of tight junction transmembrane proteins, claudin-1, -4, -7, occludin and JAM-A, in human cervical adenocarcinoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Akimoto, Taishi; Takasawa, Akira; Murata, Masaki; Kojima, Yui; Takasawa, Kumi; Nojima, Masanori; Aoyama, Tomoyuki; Hiratsuka, Yutaro; Ono, Yusuke; Tanaka, Satoshi; Osanai, Makoto; Hasegawa, Tadashi; Saito, Tsuyoshi; Sawada, NorimasaObjective. Tight junction proteins have recently been reported to be useful for distinguishing between neoplastic and non-neoplastic tissues. In this study, we evaluated the expression and localization of tight junction transmembrane proteins in human cervical adenocarcinoma and adenocarcinoma in situ (AIS), and we determined whether their expression patterns could distinguish cervical adenocarcinoma from nonneoplastic cervical glands. Methods. Fifty-five patients with cervical adenocarcinoma or AIS were included in this study. Surgical specimens were immunohistochemically stained for claudin (CLDN) -1, -4, -7, occludin, and JAM-A. Results. Significantly higher expression levels of CLDNs and JAM-A were found in cervical AIS and adenocarcinoma than in non-neoplastic glands. In cervical AIS and adenocarcinoma, localization of CLDN-1 and JAM-A was extended throughout the whole cell membranes, whereas they were predominantly expressed at the most apical cell-cell junction in non-neoplastic glands. ROC curve analysis revealed that immunoreactivities of CLDN-1 or JAM-A successfully distinguished neoplasms from nonneoplastic cervical glands with high specificity (CLDN1, 79.1%; JAM-A, 79.1%) and high sensitivity (CLDN1, 84.1%; JAM-A, 95.5%). Conclusions. As expected, there were immunohistochemical differences between cervical adenocarcinoma and non-neoplastic cervical glands by using antibodies against tight junction transmembrane proteins. These results suggest that CLDN-1 and JAM-A are potential biomarkers for cervical adenocarcinoma.
- PublicationOpen AccessThe expression of MMP-14 and microRNA-410 in FFPE tissues of human endometrial adenocarcinoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Rak, Beata; Garbicz, Filip; Paskal, Wiktor; Pełka, Kacper; Marczewska, Janina Maja; Wołosz, Dominika; Włodarski, PawełEndometrial cancer (EC) is the most common gynecological malignancy in Europe and North America. It is classified into two types exhibiting different characteristics and prognosis. Type I is an estrogen-dependent tumor, histologically classified as low grade and low stage, usually with an excellent prognosis. Type II EC is unrelated to estrogen stimulation and is characterized by a poor prognosis. MicroRNAs (miRNAs, miRs) are small non-coding RNA polynucleotides that regulate gene expression posttranscriptionally. Various dysregulations in microRNA expression are often considered to have an impact on the diagnosis, prognosis and overall survival in patients diagnosed with different types of cancers. Recent data suggest that microRNAs play an important role in the pathogenesis of EC. The aim of the study was to evaluate the involvement of matrix metaloprotease 14 (MMP-14) and microRNA-410 in formation of the EC tumor. To this end expression of MMP-14 and microRNA-410 was assessed within the cancer, transient and healthy zones in the histological sections of tumours using immunohistochemical staining and laser capture microdissection (LCM) followed by a quantitative real-time PCR. The results revealed significantly higher expression of MMP14 in the cancer tissue zone in comparison to the healthy tissue zone, as well as a lower expression of microRNA410 in the cancer zone compared with the healthy zone. This reverse correlation may suggest a regulatory role of miRNA-410 in modulating levels of MMP-14 in EC. This is the first report on such regulation in human endometrial cancer.
- PublicationOpen AccessExpression of tropomyosins in lung cancer - a potential role in carcinogenesis and its utility in a histopathological diagnosis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Okudela, Koji; Mitsui, Hideaki; Woo, Tetsukan; Kojima, Yoko; Matsumura, Mai; Arai, Hiromasa; Suzuki, Takehisa; Umeda, Shigeaki; Saito, Yuichi; Tajiri, Michihiko; Masuda, Munetaka; Kameda, Yoichi; Ohashi, KenichiWe herein analyzed the relationships between tropomyosin protein expression levels and clinicopathological factors in order to determine the significance of tropomyosins in lung cancers. Although neoplastic cells expressed different isoforms of tropomyosin, overall expression levels were lower than those in bronchial and alveolar epithelial cells. In adenocarcinomas, tropomyosin levels were markedly reduced in poorly differentiated or solid subtype carcinomas, suggesting that a loss in the expression of tropomyosins is involved in the progression of lung adenocarcinomas. The potential utility of the immunohistochemical expression of tropomyosins for a histopathological diagnosis was also investigated. The sensitivity and specificity of a loss in the expression of tropomyosins were 100% and 50%, respectively, which were superior to those for the strong expression of p53 (sensitivity 100% and specificity 44%), a conventional biomarker. An immunohistochemical examination of tropomyosins may assist in the histopathological detection of lung cancer cells in small biopsy specimens.
- PublicationOpen AccessMIR376 family and cancer(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Tekirdag, Kumsal Ayse; Akkoc, Yunus; Kosar, Ali; Gozuacik, DevrimMicroRNAs (miRNAs) are endogenous noncoding small RNAs that negatively regulate gene expression at the post-transcriptional level. They have been implicated in several fundamental biological processes including development, differentiation, apoptosis and stem cell maintenance. There is increasing evidence that microRNAs also play roles in cellular transformation and carcinogenesis by acting either as tumor suppressors or oncogenes. Recent studies introduced MIR376 as an important microRNA family for cancer formation and progression. The MIR376 family is located on human chromosome 14 and it has several members containing identical or similar seed sequences. Biological roles of family members were studied in different cancer settings, including gliomas, leukemia, breast and ovarian cancers. Furthermore, two MIR376 family members, namely MIR376A and MIR376B were implicated in the regulation of macroautophagy (autophagy herein). Since autophagy dysregulation underlies various diseases including cancer, it is essential to understand the role of the MIR376 family in this context. In this article, we summarize the miRNA-cancer connection, and review accumulating data about the involvement of the MIR376 family in cancer biology.