Histology and histopathology Vol.26, nº4 (2011)
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- PublicationOpen AccessNeonatal thymulin gene therapy in nude mice: Effects on the morphology of the pituitary corticotrope population(Murcia: F. Hernández, 2011) Martines, Eliana; Reggiani, Paula; Schwerdt, José I.; Goya, Rodolfo; Cónsole, GloriaThe integrity of the thymus during early life is necessary for a proper maturation of the neuroendocrine system, including the adrenal axis. The thymic metallopeptide thymulin seems to be a central physiologic mediator of thymus-pituitary communication. Furthermore, neonatal thymulin gene therapy has been shown to prevent the typical alterations of gonadotrophic cell number and morphology and serum gonadotropin levels in nude female mice. In the present study we assessed the impact of athymia and the effect of neonatal thymulin gene therapy on the corticotropic cell population in nude mice. The effect of thymulin administration to adult nudes on their hypothalamic content of corticotropin-releasing hormone (CRH) and the adrenal content of corticosterone was also determined. We used an adenoviral vector expressing a synthetic gene for the thymic peptide thymulin (metFTS) termed RAd-FTS. On postnatal day 1 or 2, heterozygous (nu/+) and homozygous (nu/nu) pups of both sexes received a single bilateral i.m. injection of RAd-FTS or RAd-GFP, a control vector. On postnatal day 71, mice were bled and sacrificed, and their pituitaries were immediately dissected, fixed and immunostained for corticotropin. Morphometry was performed by means of an image-analysis system. The following parameters were calculated: volume density (VD: Σ cell area/reference area), cell density (CD: number of cells/reference area), and cell surface (CS: expressed in µm2). Serum thymulin levels were measured by a bioassay, and CRH as well as corticosterone were determined by IRMA and RIA, respectively. Neonatal thymulin gene therapy in the athymic mice restored their serum thymulin levels and increased corticotrope CD, VD and CS in both control and athymic mice. Athymic mice showed only a marginal reduction in corticotrope CD, VD and CS. In these mutants hypothalamic CRH content was slightly increased, whereas adrenal corticosterone tended to be lower. Thymulin administration to adult mice tended to reverse these changes. Our results suggest a possible modulating effect of thymulin on the corticotrope population and the adrenal gland, confirming the existence of a bidirectional thymus-pituitary-adrenal axis.
- PublicationOpen AccessRole of Smad1 in diabetic nephropathy: Molecular mechanisms and implications as a diagnostic marker(Murcia: F. Hernández, 2011) Abe, H.; Matsubara, Takeshi; Arai, Hidenori; Doi, ToshioDiabetic nephropathy (DN) is the leading cause of chronic kidney failure. Moreover, DN is associated with elevated cardiovascular morbidity and mortality. DN is characterized by progressive expansion of the mesangial matrix and thickening of the glomerular basement membrane, resulting in the obliteration of glomerular capillaries. Advanced glycation endproducts (AGEs) produced as the result of hyperglycemia are known to stimulate the production of extracellular matrix (ECM) proteins, resulting in glomerulosclerosis. Exposure of cultured mesangial cells to AGEs results in a receptor-mediated upregulation of mRNA and protein secretion of type IV collagen (Col4), which is a major component of ECM. Here we review recent novel insights into the pathogenesis and diagnosis of DN, with a special emphasis on the emerging concept that diabetic glomerulosclerosis can result from activation of the signaling cascade leading to irreversible ECM overproduction. Finally, we describe signaling pathways involved in the initial change of DN and how these pathways can be manipulated for therapeutic benefit.
- PublicationOpen AccessAlterations in the dynamics of inflammation, proliferation and apoptosis in subcutaneous implants of lupus-prone mice(Murcia: F. Hernández, 2011) Campos, Paula P.; Vasconcelos, Anilton C.; Ferreira, Mônica A.N.D.; Andrade, Silvia P.Wound repair is a complex process that involves inflammation, proliferation, extracellular matrix deposition/remodeling and apoptosis. Autoimmune diseases profoundly affect the healing process. We have used histological parameters to characterize the recruitment of mast cells and the proliferative activity and apoptosis in the fibrovascular tissue induced by subcutaneous polyether-polyurethane sponge implants in lupus-prone New Zealand White (NZW) and in control Balb/c mouse strains at days 10 and 21 post implantation. Fibrovascular tissue infiltration (hematoxylin and eosin staining), mast cell number (Dominici staining) and cellular proliferation (AgNOR staining) peaked early (day 10) but collagen deposition (picrosirius red staining) and apoptosis remained high in implants of NZW mice during the experimental period. In contrast, implants of Balb/c animals showed a progressive increase in mast cell recruitment and cellular proliferation but apoptosis fell from day 10 to 21 postimplantation. This divergent response early mast cells recruitment, excessive collagen deposition and disturbed removal of apoptotic cells from the site of injury in NZW mice implies that the genotype trait of NZW mice is a determining factor in abnormal healing response.
- PublicationOpen AccessDNA repair mechanisms in mammalian germ cells(Murcia: F. Hernández, 2011) Ozturk, Saffet; Demir, NecdetMammalian germ cells encounter several types of DNA damage. This damage is almost completely repaired in a short period of time to provide the maintenance of genomic integrity. The main repair mechanisms operating in mammalian germline cells are: nucleotide excision repair (NER), base excision repair (BER), mismatch repair (MMR), DNA double strand break repair (DSBR), and post replication repair (PRR). Currently, there are relatively few publications that summarize basic information and new findings on DNA repair mechanisms used in mammalian germ cells. In the present article, we review the studies that discuss repair mechanisms operating in the female and male germ cells. We then survey some of the recent discoveries made in this field.
- PublicationOpen AccessThe primary cilium: A relevant characteristic in interstitial cells of rat duodenum enteric plexus(Murcia: F. Hernández, 2011) Junquera Escribano, Concepción; Cantarero Carmona, Irene; Luesma Bartolomé, María José; Soriano Navarro, Mario; Martínez Ciriano, Carmen; Castiella Muruzábal, Tomás; García-Verdugo, José MaríaStudies in vitro have permitted the identification of enteric neural progenitor cells. Now the question arises as to where these progenitor cells are located in vivo. The purpose of this paper is to identify possible candidate cells by means of transmission electron microscopy (TEM). We have located three interstitial cellular types around the rat duodenum myenteric plexus. Type I cells have been identified as Interstitial Cells of Cajal (ICCs). These cells present well defined ultrastructural characteristics, including the triple connexion ICnervous trunk- blood vessels. Type II cells show characteristics of immature cells, emphasizing the presence of a single cilium with the structure (9+0). To analyse this nanostructure, we have elaborated a reconstruction on ultrathin sections. The two previously described cellular types could be considered to be different functional states of the same cell. Type III cells present ultrastructural characteristics of fibroblast-like cells. This study suggests that Type II cells could be a source of neural progenitor cells.
- PublicationOpen AccessAlterations in diet consistency and variation in Hypoxia Inducible Factor-1α expression in condylar chondrocytes(Murcia: F. Hernández, 2011) Kaklamanos, Eleftherios G.; Papamitsou, Theodora; Sioga, Antonia; Economou, LouiseObjective. To investigate the expression of HIF-1α during the postnatal development of the mandibular condyle under normal and soft consistency diet conditions. Materials and Methods. Forty eight Wistar-Furth rats, aged 21 days, were divided into two groups, each being fed with either normal or soft consistency diet. Three animals from each group were sacrificed after 3, 7, 10 days (initial period), 14, 17 days (intermediate period), and 21, 24 and 28 days (final period) after the start of the experiment. Immediately after sacrifice, the mandible was excised surgically, fixed, demineralised and embedded in paraffin. Six µm thick sections were obtained, processed for conventional and immunohistochemical staining and observed in the optical microscope. HIF-1α expression was assessed semiquantitavely and graded separately for the nucleus stained cells and the cells stained exclusively in the cytoplasm. Differences in HIF-1α expression in the experimental groups were evaluated statistically. Results. HIF-1α expression was evident in the proliferative and chondroblast layers. No differences were observed between the anterior, the intermediate and the posterior parts of the condylar cartilage. In the normal consistency diet group, nuclear HIF-1α expression increased gradually until the end of the experiment. On the contrary, in the soft diet animals, nuclear HIF-1α expression increased only at the final period of the experiment. The normal diet fed animals exhibited more intense nuclear HIF-1α expression compared to cytoplasmic expression. Conclusions. HIF-1α expression in condylar chondrocytes varies under altered conditions of diet consistency
- PublicationOpen AccessMicroglia – insights into immune system structure, function, and reactivity in the central nervous system(Murcia: F. Hernández, 2011) Wirenfeldt, Martin; Babcock, Alicia A.; Vinters, Harry V.Microglia are essential cellular components of a well-functioning central nervous system (CNS). The development and establishment of the microglial population differs from the other major cell populations in the CNS i.e. neurons and macroglia (astrocytes and oligodendrocytes). This different ontogeny gives microglia unique properties. In recent years detailed studies of the microglial population have been greatly facilitated by the use of bone marrow (BM) chimeric animals. Experimental BM transplants have provided the opportunity to trace and investigate how BM cells migrate into the CNS and settle to become microglia. Furthermore various functional properties of microglia in the normal and pathological CNS are now being revealed because of combinations of BM transplantations and experimental disease models. Here, we describe some of the latest findings in microglial biology and discuss the potential for using microglia in therapeutic interventions.
- PublicationOpen AccessDecorin and biglycan expression: its relation with endothelial heterogeneity(Murcia: F. Hernández, 2011) Calabrese, Graciela C.; Gazzaniga, Silvina; Oberkersch, Roxana; Wainstok, RosaDecorin and biglycan proteoglycans play important roles in the organization of the extracellular matrix, and in the regulation of cell adhesion and migration. Given morphological and functional endothelial heterogeneity, information is needed regarding whether endothelial cells (ECs) from different vascular beds possess different profiles of proteoglycan constituents of the basement membranes. Here, we report that endothelia from different murine organs and EC lines derived thereof produce and secrete different patterns of proteoglycans. A faint colocalization between decorin and PECAM/CD31 was found on tissue sections from mouse heart, lung and kidney by immunofluorescence. Three EC lines derived from these organs produced decorin (100-kDa) and its core protein (45-kDa). Extracellular decorin recognition in culture supernatant was only possible after chondroitin lyase digestion suggesting that the core protein of secreted proteoglycan is more encrypted by glycosaminoglycans than the intracellular one. Heart and lung ECs were able to produce and release decorin. Kidney ECs synthesized the proteoglycan and its core protein but no secretion was detected in culture supernatants. Although biglycan production was recorded in all EC lines, secretion was almost undetectable, consistent with immunofluorescence results. In addition, no biglycan secretion was detected after EC growth supplement treatment, indicating that biglycan is synthesized, secreted and quickly degraded extracellularly by metalloproteinase-2. Low molecularmass dermatan sulfate was the predominant glycosaminoglycan identified bound to the core protein. ECs from different vascular beds, with differences in morphology, physiology and cell biology show differences in the proteoglycan profile, extending their heterogeneity to potential differences in cell migration capacities.
- PublicationOpen AccessQuantitative analysis of vessels with smooth muscle layer in astrocytic tumors: correlation with histological grade and prognostic significance(Murcia: F. Hernández, 2011) Sato, Shinya; Sato, Yuichiro; Hatakeyama, Kinta; Marutsuka, Kousuke; Yamashita, Atsushi; Takeshima, Hideo; Asada, YujiroAngiogenesis plays an important role in the progression of astrocytic tumors and its evaluation is a major prognostic factor. Although the form of proliferating vessels ranges from fine capillaries to welldeveloped vascular structures with a smooth muscle layer, the characteristics of vascular smooth muscle cells (SMCs) are not understood in detail. We therefore examined the density, size and shape of tumor vessels, as well as CD34-immunoreactive (CD34-Vs) or α-smooth muscle actin-immunoreactive (SMA-Vs) vessels in 46 primary astrocytomas (grade II diffuse astrocytomas, n=11, grade III anaplastic astrocytomas, n=15, grade IV glioblastomas, n=20) and in normal brain tissues from 10 autopsies. We also examined the expression of high molecular weight caldesmon (h-CD, a marker of the contractile phenotype of smooth muscle) and of plateletderived growth factor receptor ß (PDGFR-ß). The SMAVs were significantly more dense and larger in grade IV than grade III, whereas those of CD34-Vs did not differ between grade III and IV. Changes in the shape of CD34-Vs and SMA-Vs correlated with histological grading. The expression of h-CD was reduced, whereas that of PFGFR-ß was increased in high gradeastrocytomas. Kaplan-Meier analysis indicated that the density of SMA-Vs, the size of both CD34 and SMA-Vs and PDGFR-ß expression were significant prognostic factors. These findings suggest that SMA-Vs are significantly associated with the progression of astrocytomas and that these vessels provide useful information for the histological diagnosis and survival of patients with these types of brain tumors.
- PublicationOpen AccessThe use of slaughterhouse-obtained small intestinal tissue as control material in histological studies should be applied with prudence(Murcia: F. Hernández, 2011) De Ceulaer, K.; Van Ginneken, C.; Delesalle, C.; Van Brantegem, L.; Deprez, P.; Weyns, A.This study aimed to evaluate the reliability of slaughterhouse-obtained small intestinal tissue as control material in equine colic research where molecular stress responses in small intestinal tissue are investigated. For this purpose, small intestinal samples from colic horses were collected during surgery or immediately after euthanasia at the oral border of strangulation resection sites and routinely processed for histopathology (i.c. rinsed with 4°C Krebs’ solution, fixated overnight with 4% neutral buffered formaldehyde (FH) at room temperature). Control samples consisted of pieces of mid-jejunum, collected at the slaughterhouse and routinely processed for histopathology under 4 different conditions. The 4 conditions differed with regard to incubation and fixation temperature and whether or not oxygenated Krebs’ solution was used. Histological scoring revealed that slaughterhouse samples had a higher mean lesion score (P<0.001) than colic samples. In addition, more slaughterhouse samples had a higher mean inflammation score than colic samples (P=0.001). The inflammatory cells in the small intestine consisted mostly of eosinophils and as such were very suggestive for parasitic infestation. Hypoxia-inducible factor-1α (HIF1α) nuclear immunoreactivity was more pronounced in slaughterhouse tissue, probably as a result of the delay between slaughter and sampling (P=0.034). The histopathological score (P=0.291), the inflammation score (P=0.248) and the HIF1α nuclear immunoreactivity (P=0.538) did not differ between the different collection protocols. It is concluded that slaughterhouseobtained small intestinal tissue shows distinct alterations and that its use as control tissue when evaluating molecular stress responses should be applied with prudence.
- PublicationOpen AccessTranscription factor snail1 expression and poor survival in pharyngeal squamous cell carcinoma(Murcia: F. Hernández, 2011) Jouppila-Mättö, Anna; Tuhkanen, Hanna; Soini, Ylermi; Pukkila, Matti; Närkiö-Mäkelä, Mervi; Sironen, Reijo; Virtanen, Ismo; Mannermaa, Arto; Kosma, Veli-MattiSnail1, a key regulator of epithelialmesenchymal transition (EMT), plays an important role in tumour progression. Previous studies of snail1 have mainly focused on the epithelial tumour cells. The objective of this study was to evaluate the expression of snail1 protein in endothelial cells, stromal myofibroblasts and malignant epithelial cells of pharyngeal squamous cell carcinomas (PSCC), as well as its relation to clinicopathological features and survival. One hundred and ten tissue microarray samples were analyzed for snail1 expression using immunohistochemistry. In endothelial cells snail1 expression was observed in 51 (48%) of 107 cases and it predicted reduced disease specific survival (DSS) (p=0.009). In 49 (46%) tumour samples snail1 immunostaining was detected in stromal myofibroblasts and there was a tendency to poorer DSS in that group (p=0.067). Snail1 expression in endothelial cells and stromal myofibroblasts is also associated with hypopharyngeal tumours (p=0.01 and p=0.038 respectively), increasing T category (T3-4) (p=0.005, p=0.037 respectively) and poorer general condition of the patient (Karnofsky performance status score <70; p=0.029, p=0.039 respectively). Moreover endothelial expression correlated with advanced stage (III-IV) (p=0.005) and poorer differentiation (grade 2-3; p=0.012). In malignant epithelial cells snail1 immunostaining was detected in 75 of 110 cases (68%). Expression of the protein was more common in hypopharyngeal tumours (p=0.044). Snail1 positive tumours associated with a lower Karnofsky performance status score (p=0.039) and regional failure (p=0.042). Our findings indicate that snail1 protein expression in endothelial cells and to some extent also in tumour stromal myofibroblasts seems to be a predictor of poor survival in PSCC. The presence of snail1 protein in tumour microenvironment rather than in malignant epithelial tumour cells may induce tissue remodelling and tumour progression.
- PublicationOpen AccessGenetic alterations in pulmonary epithelioid hemangioendothelioma and epithelioid angiosarcoma(Murcia: F. Hernández, 2011) Cao, Y.; Zou, S.M.; Zhang, K.T.; Lu, N.; Liu, Y.; Feng, L.; Wen, P.; Han, N.J.; Lin, D.M.Epithelioid hemangioendothelioma (EHE) is a low-to-intermediate-grade vascular tumor that occurs in many organs, and epithelioid angiosarcoma (EA) is a subtype of angiosarcoma that is associated with highgrade malignancy. These two types of tumors have different forms of biological behavior. Pulmonary epithelioid hemangioendothelioma (PEH) and epithelioid angiosarcoma (PEA) are both very rare, and genetic studies on them are extremely limited. We examined and compared the cytogenetic characteristics of these two types of lung tumors in two patients utilizing the Array-Comparative Genomic Hybridization (Array-CGH) method. Considerable differences in the cytogenetic characteristics were observed between the two types of tumors. Small fragment gains (<10 MB) were dominant in PEH, whereas large fragment gains and deletions (>10 MB) were dominant in PEA. Some large fragment alterations, such as gains in chromosomes 19q and 19p, and deletions in chromosomes 9p and 13q, involved over half of a chromosome arm. PEH and PEA showed great cytogenetic differences; therefore, further genetic studies on these two types of tumors are warranted.
- PublicationOpen AccessOverexpression of Aquaporin-1 in lung adenocarcinomas and pleural mesotheliomas(Murcia: F. Hernández, 2011) López-Campos, José Luis; Sánchez Silva, Rocío; Gómez Izquierdo, Lourdes; Márquez, Eduardo; Ortega Ruiz, Francisco; Cejudo, Pilar; Barrot Cortés, Emilia; Toledo Aral, Juan José; Echevarría, MiriamAquaporin-1 (AQP1) is the main water channel responsible for water transport through many epithelia and endothelia. The latest evidence pointed toward an important role of this protein also in gas permeation, angiogenesis, cell proliferation and migration. In the present work we studied the expression of AQP1 by immunohistochemical staining of 92 lung biopsies from patients diagnosed with a pleuropulmonary tumor (71 lung and 21 pleural neoplasms). AQP1 expression was analyzed comparing the results among the different histological patterns and against 9 control cases (5 parenchyma and 4 healthy pleura). Clear staining of AQP1 was detected in 39 of the 92 tumors analyzed. In parenchyma, AQP1 was more frequently detected in primary lung adenocarcinomas (55%, P<0.001); in contrast, small cell carcinomas were the least AQP1 expressive tumors studied (93% of negative staining, P<0.05). Carcinomas analyzed in pleura (mesotheliomas and metastatic adenocarcinomas) also revealed strong expression of AQP1. High expression of this protein was detected in small capillaries in areas near or surrounding the tumor, and novel intense AQP1 immunostaining was detected over thicker alveolar walls in alveoli inside or next to the tumoral tissue regardless of the tumor type. An important role of AQP1 in tumor angiogenesis is sustained by the abundant expression of this protein in the endothelia of tumor capillaries. Further studies are necessary to elucidate the potential pathophysiological role of this protein in pleuropulmonary neoplasms.