Histology and histopathology Vol.25, nº1 (2010)
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- PublicationOpen AccessGlycoconjugates within the oviduct and their functional significance with special reference to marsupials(Murcia : F. Hernández, 2010) Chapman, Jamie A.; Chuah, M.I.; Breed, William G.In placental (eutherian) mammals, a number of important events take place within the oviduct including the pre-fertilisation maturation of gametes (including sperm storage), sperm-egg interactions, egg activation and early embryonic development. Many of these events involve interactions of glycoconjugates; both on the surface of the gametes and with the secretions of the oviductal epithelium and these have best been studied in eutherian mammals. In marsupials, however, while the oviduct is known to produce the extracellular egg coat, the mucoid layer, that comes to surround the zona pellucida, its role in the maturation of gametes is only now being elucidated, particularly in the oocyte. This review emphasises what is known of the structure and function of the oviduct and its secretions in marsupials and briefly compares it with data from eutherians. In particular, knowledge of oviductal glycoconjugates in the structure of the post-ovulatory oocyte and its vestments around the time of fertilisation in Australian marsupials is outlined.
- PublicationOpen AccessNicotinamide N-Methyltransferase upregulation correlates with tumour differentiation in oral squamous cell carcinoma(Murcia : F. Hernández, 2010) Emanuelli, Monica; Santarelli, Andrea; Sartini, Davide; Ciavarella, Domenico; Rossi, Valentina; Pozzi, Valentina; Rubini, C.; Lo Muzio, L.We investigated expression levels of Nicotinamide N-Methyltransferase (NNMT), an enzyme involved in the biotransformation of many drugs and xenobiotic compounds, in oral squamous cell carcinoma (OSCC). Measurements were performed by immunohistochemistry and the relationship between tumour characteristics and NNMT levels in OSCC was studied to evaluate the effectiveness of NNMT as a prognostic marker in squamous cell carcinoma of the oral cavity. In conclusion, the present study suggests that NNMT may have potential as a biomarker and as a therapeutic target for OSCC.
- PublicationOpen AccessThe amniotic membrane as a source of stem cells(Murcia : F. Hernández, 2010) Insausti, Carmen L.; Blanquer Blanquer, Miguel; Bleda, Patricia; Iniesta, Paqui; Majado Martínez, Mª Juliana; Castellanos Escrig, Gregorio; Moraleda Jiménez, José MaríaCellular therapy has emerged as a new potential tool for curing a wide range of degenerative diseases and tissue necrosis. Embryonic stem cells possess potential for differentiation into a wide range of cell lineages, but the ethical issues associated with establishment of this human cell line have to be resolved prior to any use. The bone marrow (BM) is the usual source of adult stem cells for hematopoietic stem cell transplants and cellular therapy, but the BM harvest is a surgical procedure that requires general anesthesia or sedation, and there seems to be a reduction of the proliferative potential and differentiation capacity of the marrow mesenchymal stem cells in older donors. For these reasons there is an increasing interest in other sources of stem cells from adult and fetal tissues. The amniotic membrane (AM) or amnion is a tissue of particular interest because its cells possess characteristics of stem cells with multipotent differentiation ability, and because of low immunogenicity and easy procurement from the placenta, which is a discarded tissue after parturition, thus avoiding the current controversies associated with the use of human embryonic stem cells. Therefore, amniotic membrane has been proposed as a good candidate to be used in cellular therapy and regenerative medicine.
- PublicationOpen AccessThe early endosome, a busy sorting station for proteins at the crossroads(Murcia : F. Hernández, 2010) Jovic, Marko; Sharma, Mahak; Rahajeng, Juliati; Caplan, SteveEndocytosis marks the entry of internalized receptors into the complex network of endocytic trafficking pathways. Endocytic vesicles are rapidly targeted to a distinct membrane-bound endocytic organelle referred to as the early endosome. Despite the existence of numerous internalization routes, early endosomes (EE) serve as a focal point of the endocytic pathway. Sorting events initiated at this compartment determine the subsequent fate of internalized proteins and lipids, destining them either for recycling to the plasma membrane, degradation in lysosomes or delivery to the trans-Golgi network. Sorting of endocytic cargo to the latter compartments is accomplished through the formation of distinct microdomains within early endosomes, through the coordinate recruitment and assembly of the sorting machinery. An elaborate network of interactions between endocytic regulatory proteins ensures synchronized sorting of cargo to microdomains followed by morphological changes at the early endosomal membranes. Consequently, the cargo targeted either for recycling back to the plasma membrane, or for retrograde transport to the trans-Golgi network, localizes to newly-formed tubular membranes. With a high ratio of membrane surface to lumenal volume, these tubules effectively concentrate the recycling cargo, ensuring efficient transport out of the EE. Conversely, receptors sorted for degradation cluster at the flat clathrin lattices involved in invaginations of the limiting membrane, associating with newly formed intralumenal vesicles. In this review we will discuss the characteristics of early endosomes, their role in the regulation of endocytic transport, and their aberrant function in a variety of diseases.
- PublicationOpen AccessThe relative effects of severe burn injury and pre- and post-natal protein deprivation on mandibular condyle morphology(Murcia : F. Hernández, 2010) Caixeta de Oliveira, Bruna Cecília; de Oliveira, Flávia; Terra Martini, Dorival; Duarte Prisco, Cleide Rosana; da Silva Riguetti, Marta Maria; Aparecido Liberti, Edson; de Campos Boldrini, SilviaThe mandible has a mixed embryological origin, and its growth is associated with the secondary cartilage of the condyle process (CP). In this area, growth depends on an array of intrinsic and extrinsic factors that influence protein metabolism. In the present study, we used an adolescent rat model to evaluate the growth and development of the CP under conditions of pre- and postnatal protein deficiency, combined with or without the stress of severe burn injury (BI). We found that protein deficiency severely undermined the growth of the CP, by altering the thickness of its constituent layers. BI is also capable of affecting CP growth, although the effect is less severe than protein deficiency. Interestingly, the summed effect of protein deficiency and BI on the CP is less severe than protein deficiency alone. A possible explanation is that the increased carbohydrates in a hypoproteic diet stimulate the production of endogenous insulin and protein synthesis, which partially compensates for the loss of lean body mass caused by BI.
- PublicationOpen AccessDistribution of components of basal lamina and dystrophin-dystroglycan complex in the rat pineal gland, differences from the rat pineal gland: differences from the brain tissue and between the subdivisions of the gland(Murcia : F. Hernández, 2010) Bagyura, Zsolt; Pócsai, Károly; Kálmán, MihályThe pineal gland is an evagination of the brain tissue, a circumventricular neuroendocrine organ. Our immunohistochemical study investigates basal lamina components (laminin, agrin, perlecan, fibronectin), their receptor, the dystrophin-dystroglycan complex (ß-dystroglycan, dystrophin utrophin), aquaporins (-4,-9) and cellular markers (S100, neurofilament, GFAP, glutamine synthetase) in the adult rat corpus pineale. The aim was to compare the immunohistochemical features of the cerebral and pineal vessels and their environment, and to compare their features in the distal and proximal subdivisions of the so-called ’superficial pineal gland’. In contrast to the cerebral vessels, pineal vessels proved to be immunonegative to α1-dystrobrevin, but immunoreactive to laminin. An inner, dense, and an outer, loose layer of laminin as two basal laminae were present. The gap between them contained agrin and perlecan. Basal lamina components enmeshed the pinealocytes, too. Components of dystrophin-dystroglycan complex were also distributed along the vessels. Dystrophin, utrophin and agrin gave a ’patchy’ distribution rather than a continuous one. The vessels were interconnected by wing-like structures, composed of basal laminacomponents: a delicate network forming nests for cells. Cells immunostained with glutamine synthetase, S100- protein or neurofilament protein contacted the vessels, as well as GFAP- or aquaporin-immunostained astrocytes. Within the body a smaller, proximal, GFAP-and aquaporin-containing subdivision, and a larger, distal, GFAP-and aquaporin-free subdivision could be distinguished. The vascular localization of agrin and utrophin, as well as dystrophin, delineated vessels unequally, preferring the proximal or distal end of the body, respectively.
- PublicationOpen AccessThe role of cancer stem cells and the side population in epithelial ovarian cancer(Murcia : F. Hernández, 2010) Fong, Miranda Y.; Kakar, S.S.Ovarian cancer is the most lethal cancer of the female reproductive tract, accounting for ~15,000 deaths per year according to the National Cancer Institute and American Cancer Society. This review article covers risk factors for the development of ovarian cancer, current detection strategies, prognostic markers, treatment strategies, etiology of tumorigenesis, and ovarian somatic stem cells. While the etiology of ovarian cancer is still unknown, several theories have been proposed as the mechanism of carcinogenesis. One theory states that the surface epithelium undergoing invagination and forming inclusion cysts that are exposed to growth factors and cytokines. The “gonadotropin theory” has also been proposed. Other reigning models for tumorigenesis include the stochastical model where a distinct population of cells acquires somatic mutations leading to metastasis, and the hierarchical model where the tumor is initiated by cancer stem cells (CSCs). CSCs isolated from primary tumors have the ability to regenerate the tumor and reconstitute the original tumor phenotype with as few as 100 cells. CSCs from ovarian carcinomas display the cell surface markers CD44+CD117+CD133+. CSCs are also thought to account for chemotherapy resistance through the expression of highly selective transporters ABCG2 and MDR1 and activation of TLR4/MyD88. The side population has been characterized by their ability to efflux lipophilic substrates, including the dye Hoechst 33342 and many chemotherapy agents. This ability has been attributed to the expression of the transporters ABCG2 and MDR1.
- PublicationOpen AccessExpression of OAT1 and OAT3 in differentiating proximal tubules of the mouse kidney(Murcia : F. Hernández, 2010) Hwang, Jin-Sun; Park, Eun-Young; Kim, W.J.; Yang, Chul-Woo; Kim, JinOrganic anion transporter 1 (OAT1) and OAT3 in the proximal tubules (PT) of the kidney play important roles in the elimination of harmful endogenous compounds and xenobiotics from the body. We investigated the temporal and spatial expression of OAT1 and OAT3 in the differentiating PT in mouse kidney. Ontogenic expression of OAT1 and OAT3 was investigated by immunohistochemical analysis. The S1, S2, and S3 segments of the PT were identified using antibodies to aquaporin 1 (AQP1), Na+-HCO3 – cotransporter 1 (kNBC1), and AQP4. OAT1 immunoreactivity was first detected at PT in the inner cortex of 15-day-old fetuses (F15) and in the outer cortex of 7-day old pups. OAT3 was first observed in the distal tubule of F14 and in S2 segment of the PT of F16 and in S1 and S3 segments around the time of birth; expression increased through postpartum day 21. The ontogenic pattern of expression of OAT1 and OAT3 in the differentiating PT suggests that both transporters may function in the S2 segment in the fetus, but not until after birth in S1 and S3 segments.
- PublicationOpen AccessExpression of Reg IV and Hath1 in neuroendocrine neoplasms(Murcia : F. Hernández, 2010) Heiskala, Kukka; Arola, Johanna; Heiskala, Marja; Andersson, Leif C.Reg IV (RELP), a Regenerating protein family member, is constitutively expressed in neuroendocrine cells of the intestinal mucosa. The helixloop-helix transcription factor Hath1 is the human homologue of murine Math1, which regulates the embryonic differentiation of neural and intestinal secretory lineage cells. Hath1 is constitutively expressed in a subset of mature secretory gastrointestinal cells. We investigated by immunohistochemistry the expression of Reg IV and Hath1 in 63 neuroendocrine tumors. Intestinal neuroendocrine neoplasms showed coexpression of Reg IV and Hath1, as did parathyroidal and Merkel cell tumors. Lung small-cell carcinoma and gastric mucocellular carcinoma expressed only Reg IV. Pancreatic islet-derived tumors, pheochromocytomas, and paragangliomas expressed only Hath1. Lymph node and liver metastases retained the tissue-specific expression patterns. These distinct expression profiles may be useful for differential diagnostics of metastatic lesions of neuroendocrine tumors. The dissimilar expression patterns suggest that the proteins belong to different signaling pathways and are activated at different stages of neuroendocrine differentiation. Local Reg IV expression may be influenced by the growth factors bFGF and HGF and/or their receptors CD138 and c-met, which were found to co-localize with Reg IV in intestinal neuroendocrine tumors.
- PublicationOpen AccessClaudins in human cancer, A review(Murcia : F. Hernández, 2010) Ouban, Abderrahman; Ahmed, A.Claudins are tight junction proteins that are critical for the sealing of cellular sheets and controlling paracellular ion flux. The claudin family of proteins is composed of at least 24 closely related transmembrane proteins, most of them are well characterized at the gene and protein levels. The claudins are present in variety of normal tissues, hyperplastic conditions, benign neoplasms, and cancers that exhibit epithelial differentiation. Loss of claudins expression has also been reported in several malignancies as well. Differential expression of various members of the claudins family in cancers can be used in confirming the histologic identity of certain cancers and excluding others. Examples include the use of immunohistochemical detection of claudins to differentiate between oncocytoma and chromophobe renal cell carcinoma, endometrial endometrioid carcinoma and seropapillary carcinoma, mesothelioma and metastatic adenocarcinoma, hepatocellular and biliary tract carcinomas, and between intestinal-type and diffuse-type gastric carcinoma. Expression of certain claudins can also be used as markers that can predict patient’s prognosis. Thus, it seems that attempts to identify expression claudins in cancers are becoming increasingly useful in histologic diagnosis of tumors as well as means to assess patient’s prognosis.
- PublicationOpen AccessExpression of claudin-1, -3, -4, -5 and -7 proteins in low grade colorectal carcinoma of canines(Murcia : F. Hernández, 2010) Jakab, Cs.; Rusvai, M.; Gálfi, P.; Szabó, Z.; Szabára, Á.; Kulka, J.The aim of the present study was to characterise the expression pattern of claudin-1, -3, -4, -5 and -7 tight junction proteins in canine normal colorectum and in the low-grade, tubulopapillary colorectal carcinoma in canines. Methods and results: The biopsy samples included 10 canine normal colorectal tissues and 20 canine low grade colorectal carcinomas (CLGCCs). The canine normal colorectal mucosa was negative for claudin-1. Claudin-1 was detected as a non-diffuse intense membrane labelling of neoplastic epithelial cells in low grade colorectal cancer in canines. Fifty five per cent of all tumours showed a weak cytoplasmic pattern of staining for claudin-1 protein. The normal colorectal mucosa showed diffuse punctate positivity for claudin-3. Claudin-3 was detected as an intense lateral membrane labelling of tumour cells in CLGCCs. Claudin-4 expression in surface and crypt epithelial cells of the intact colorectal mucosa in canines was punctate. Claudin-4 molecule was detected as a lateral membrane labelling of neoplastic cells in CLGCCs. The epithelium of the CLGCCs and the low grade colorectal carcinoma were negative for claudin-5. The surface and crypt epithlial cells of the canine normal colorectal mucosa showed a diffuse lateral membranous pattern of staining for claudin-7. Claudin-7 molecule was detected as an intense membrane labelling of neoplastic cells in CLGCCs. Seventy per cent of all tumours showed weak cytoplasmic positivity for claudin-7. Conclusion: Consequently, we hypothesize that claudin-1 plays a role in the progression of CLGCCs. Further functional studies are needed to clarify the biological role of the mislocalization of the claudin-1 molecule from cell membrane to the cytoplasm in CLGCCs. Lower claudin-4 expression suggests that reduced expression of claudin-4 molecule may lead to cellular disorientation, detachment and invasion of CLGCCs. Further functional studies are needed to clarify the biological role of overexpression and mislocalisation of claudin-7 in CLGCCs.
- PublicationOpen AccessMorphological and biochemical patterns in skeletal muscle apoptosis(Murcia : F. Hernández, 2010) D’Emilio, A.; Biagiotti, L.; Burattini, S.; Battistelli, M.; Canonico, B.; Evangelisti, C.; Ferri, P.; Papa, S.; Martelli, A.M.; Falcieri, E.Some neuromuscular disorders, such as Duchenne muscular dystrophy, hereditary inclusion body myopathy, malignant hyperthermia, alcoholic myopathy and mitochondrial myopathies are characterized by oxidative stress and loss of muscle fibres due to apoptosis. In this study we have analyzed muscle cell death in vitro utilizing C2C12 myoblasts and myotubes, inducing apoptosis by means of UVB irradiation. C2C12 cells were analysed by scanning and transmission electron microscopy (SEM, TEM) as well as by TUNEL reaction. DNA analysis was performed by gel electrophoresis and flow cytometry. MitoTracker red CMXRos and JC-1 fluorescent probes were also used to study mitochondrial behavior. Finally, caspase activity was investigated by means of Western blot, while caspase-9 and -3 inhibitor effects by means of SEM. SEM showed the typical membrane blebbing while TEM revealed the characteristic chromatin condensation. The TUNEL reaction presented a certain positivity too. Apoptotic and non-apoptotic nuclei in the same myotube were identified both by TUNEL and TEM. Gel electrophoresis never showed oligonucleosomal DNA fragmentation, in agreement with the cell cycle analysis performed by flow cytometry which did not reveal a sharp subdiploid peak. Mitochondrial response to UVB was later investigated and a decrease in mitochondrial functionality appeared. Caspase-9 and -3 cleavage, and, consequently, the activation of the caspase cascade, was also demonstrated by Western blot. Moreover a decrease in apoptotic cell number was noted after caspase-9 and-3 inhibitor treatment. All these results indicated that UVB irradiation induces apoptosis, both in myoblasts and in myotubes, the second being more resistant. DNA fragmentation, at least the nucleosomic type, does not occur. A certain double-strand cleavage appears in TUNEL analysis, as well as characteristic ultrastructural changes in chromatin.
- PublicationOpen AccessNo loss of melanopsin-expressing ganglion cells detected during postnatal development of the mouse retina(Murcia : F. Hernández, 2010) González-Menéndez, Irene; Contreras, Felipe; Cernuda-Cernuda, R.; García-Fernández, J.M.Melanopsin, an opsin protein expressed in mammalian retinal ganglion cells (RGCs), makes them responsive to light. Such photosensitive RGCs form the retinohypothalamic tract (RHT) that provides signals to the suprachiasmatic nucleus (SCN), the master regulator of circadian rhythms. The SCN is adjusted daily to the environmental day/night cycle by signal inputs incoming from the RHT. In the present work we have studied, using immunohistochemistry techniques, the types and number of cells which expressed melanopsin during the postnatal development of pigmented C3H/He mice maintained in a standard daily cycle (12-h light / 12-h dark). Our results clearly show for the first time that the retina maintains a rather constant number of melanopsinexpressing RGCs from the first postnatal day and, thus, demonstrate that no loss of these photosensitive cells occurs during postnatal development. This supports the general idea that the non-image-forming system, in which these cells are involved, is functional at the very early postnatal stage.