Histology and histopathology Vol.27, nº 8 (2012)

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Open Access
The involvement of the spleen during chronic phase of Schistosoma mansoni infection in galectin-3-/- mice
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Brand, Camila; Oliveira, Felipe L.; Takiya, Christina M.; Palumbo Jr, Antonio; Hsu, Daniel K.; Liu, Fu-Tong; Borojevic, Radovan; Chammas, Roger; El-Cheikh, Márcia C.
Schistosoma mansoni synthesizes glycoconjugates which interact with galectin-3, eliciting an intense humoral immune response. Moreover, it was demonstrated that galectin-3 regulates B cell differentiation into plasma cells. Splenomegaly is a hallmark event characterized by polyclonal B cell activation and enhancement of antibody production. Here, we investigated whether galectin-3 interferes with spleen organization and B cell compartment during chronic schistosomiasis, using wild type (WT) and galectin-3-/- mice. In chronically-infected galectin-3-/- mice the histological architecture of the spleen, including white and red pulps, was disturbed with heterogeneous lymphoid follicles, an increased number of plasma cells (CD19-B220-/lowCD138+) and a reduced number of macrophages (CD19-B220-Mac-1+CD138-) and B lymphocytes (CD19+B220+/highCD138-), compared with the WT infected mice. In the absence of galectin-3 there was an increase of annexin-V+PI- cells and a major presence of apoptotic cells in spleen compared with WT infected mice. In spleen of WT infected mice galectin-3 was largely expressed in lymphoid follicles and extrafollicular sites. Thus, we propose that galectin-3 plays a role in splenic architecture, controlling distinct events such as apoptosis, macrophage activity, B cell differentiation and plasmacytogenesis in the course of S. mansoni infection
Open Access
Von Hippel-Lindau Disease (VHL): A need for a murine model with retinal hemangioblastoma
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Park, Stanley; Chan, Chi-Chao
Von Hippel-Lindau (VHL) disease is a highly penetrant autosomal dominant systemic malignancy that gives rise to cystic and highly vascularized tumors in a constellation of organs. Patients with VHL disease commonly present with hemangioblastomas in the central nervous system and the eye while other manifestations include pheochromocytoma, clear cell renal cell carcinoma, endolymphatic sac tumors of the middle ear, pancreatic cystadenomas, epididymal and broad ligament cystadenomas. Animal models inactivating the VHL gene product in various organ tissues have been constructed over the past 15 years to parse its HIF-associated mechanisms and its link to tumorigenesis. These models, despite advancing our understanding the molecular role of VHL, are by and large unable to recapitulate the more common features of human VHL disease. Up to date, no model exists that develop retinal hemangioblastomas, the most common clinical manifestation. The purpose of this review is: (1) to discuss the need for an ocular VHL model, (2) to review the animal models that recapitulate clinical VHL disease and (3) to propose potential mechanisms of tumorigenesis for the development of ocular VHL
Open Access
Multilocular cystic renal cell carcinoma with focus on clinical and pathobiological aspects
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Kuroda, Naoto; Ohe, Chisato; Mikami, Shuji; Inoue, Keiji; Nagashima, Yoji; Cohen, Ronald J.; Pan, Chin-Chen; Michal, Michal; Hes, Ondrej
Multilocular cystic renal cell carcinoma (MCRCC) accounts for approximately 1 to 2% of all renal tumors. This tumor is currently classified as a subtype of clear cell RCC. Clinically, the majority of these tumors are incidentally found. Macroscopically, the tumor is well demarcated and consists of various-sized cysts. The fibrous septa are generally thin and there is no discernible expansile nodule. Microscopically, the cyst walls are lined with tumor cells with clear to occasionally slightly eosinophilic cytoplasm. The Fuhrman nuclear grade is generally low and usually corresponds to grade 1. The deletion of chromosome 3p was identified in most tumors using FISH analysis and VHL gene mutation was identified in 25% of MCRCC. As MCRCC generally exhibits a low stage of TNM classification, the great majority of these tumors have a favorable clinical course. To date, there are no reports of metastasis, vascular invasion or sarcomatoid change in MCRCC. Accordingly, nephron sparing surgery is first recommended as a therapeutic strategy
Open Access
Neuroma under the fifth metatarsal head. A retrospective study
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Valero, J.; Gallart, J.; González, D.; Agustín, L.; Marquina, R.; Deus, J.; Lahoz, M.
This retrospective study was carried out over 83 surgical cases at the distal portion of the fifth metatarsal, compromising the treatment of tailor’s bunion, fifth metatarsal overload and the concomitant presence of both pathologies in some cases. Neuromas were founded under the fifth metatarsal head in 18 of the cases studied (21.7%). The results look at whether if there is an association between different fifth metatarsal pathologies and the presence of neuromas and found a significant association between the appearance of neuromas in patients with the same metatarsal overload, especially if it is accompanied by a tailor’s bunion pathology
Open Access
Development of the human foreskin during the fetal period
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Alves Favorito, Luciano; Balassiano, Carlos Miguel; Silva Costa, Waldemar; Barcellos Sampaio, Francisco José
Aims: Foreskin development begins at twelfth gestational week through a circular invagination of the ectoderm in the glandular periphery that grows ventrally and totally involves the glans around the twentieth gestational week. Studies of foreskin formation chronology and its histological constituents in human fetuses are rare. The objective of this study is to analyze foreskin development during the second trimester of the human fetal period. Methods: We studied twelve well-preserved human fetuses between thirteen and nineteen weeks post conception (WPC), according to the foot length criterion. The fetuses’ weight ranged from 70 to 340 g and the crown-rump length from 11 to 18.5 cm. Their penises were formalin-fixed, paraffin-embedded and cut into 5 micrometers sections. Hematoxylin and eosin, Van Gieson solution, Gomori trichrome and Weigert staining were used. Results: The glans was partially covered by the foreskin in the fetus at 13 WPC and almost completely covered by the foreskin in fetuses at 16 WPC and 17 WPC. The complete foreskin was formed only in the fetuses at 18 and 19 WPC, in which the foreskin totally covered the glans. In all the fetuses studied we observed the presence of preputial lamella and a large amount of mesenchymal tissue between the foreskin and glans. Conclusion: The chronology of foreskin formation in the second gestational trimester is well documented in our article. It is a fast process that lasts around five weeks and is coordinated with penile urethra formation
Open Access
Telocytes, a distinct type of cell among the stromal cells present in the lamina propria of jejunum
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Cretoiu, D.; Cretoiu, Sanda M.; Simionescu, Anca A.; Popescu, L.M.
Conventionally, cells described in the stroma of the intestinal wall are fibroblasts/fibrocytes, mast cells, plasma cells, eosinophils, macrophages and, interstitial cells of Cajal (ICCs), the latter being considered as the pacemakers of gastrointestinal rhythmicity. Recently, a new type of stromal cell called telocyte (TCs) was found in various cavitary and non-cavitary organs (www.telocytes.com). We show here direct electron microscopical evidence for the presence of TCs in the lamina propria of rat jejunum just beneath the epithelial layer of the mucosal crypts and in between the smooth muscle cells (SMCs) of muscularis mucosae. TCs are characterized by: several very long (tens to hundreds of µm) prolongations called telopodes (Tps). Tps (with caliber below the resolving power of light microscopy) display podomeres (thin segments ≤0.2 µm) and podoms (dilations accommodating caveolae, mitochondria, and endoplasmic reticulum). Tps present dichotomous branching and form a three dimensional network close to immune cells, SMCs or nerve bundles. TCs could play a role in intercellular signaling and control of local tissue homeostasis
Open Access
HER2 status determination using RNA-ISH - a rapid and simple technique showing high correlation with FISH and IHC in 141 cases of breast cancer
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Alba, Javier; Gutierrez, Javier; Coupe, Victoria Mary; Fernández, Beatriz; Vázquez-Boquete, Angel; Alba, Jesús; Forteza, Jeronimo; García-Caballero, Tomás
Aims: the assessment of the human epidermic growth factor receptor 2 (HER2) is currently performed in most laboratories using two techniques: Fluorescence in situ hybridisation (FISH) and inmunohistochemistry (IHC), and novel methodology is being investigated continuously in the assessment of HER2, such as SISH, CISH, DNA chips, ELISA or real time PCR to make assessment easier, faster or more accurate. RNA-ISH (RNA in Situ Hybridisation) is a new technique designed to detect mRNA expression levels, conducted by light microscope without the need for counting or grading systems in a total processing time of 4 hours. This study aims to determine if RNA-ISH is a viable and effective technique and a possible alternative to the currently used techniques by analysing and comparing genetic amplification (FISH) and protein levels (IHC) with mRNA over-expression (RNA-ISH) in 141 cases of breast cancer. Results: This study demonstrated a 96.5% concordance between over-expression of HER2 as determined by RNA-ISH and gene amplification as determined by FISH. The relationship between RNA-ISH-evaluated and IHC-evaluated over-expression was equally well reflected with a 95.2% concordance. Importantly, a considerable reduction in processing and evaluation time was achieved of only 4 hours. Conclusions: We conclude that the probe developed for RNA-ISH represents a viable, effective possible alternative to FISH and IHC for analysing HER2 status in primary breast tumours
Open Access
Clonal relationship of relapsing lymphoid neoplasms
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Obermann, E.C.; Dirnhofer, S.; Tzankov, A.
Lymphomas encompass a broad spectrum of neoplasias. Traditionally it has been assumed that recurrent neoplasia, especially lymphoma, represents a relapse of the original clone. However, this concept has been challenged. Here we present an overview of novel perceptions regarding the clonal relationship of relapsing lymphoid neoplasms, i.e. precursor cell acute lymphoblastic lymphoma/leukemia (ALL), so called non-Hodgkin lymphomas (NHL) and classical Hodgkin lymphoma (cHL) and discuss the potential implications of these findings. In ALL, approximately 10% of “relapses” were found to be clonally unrelated to the original disease. In NHL, small series and case reports showed the occurrence of meta- or synchronous lymphoid malignancies, which were of different clonal origin. In cHL, there is evidence that both early and late “relapses” may constitute to a certain proportion a novel neoplasm of different clonal origin too. These findings warrant further investigations in order to verify and strengthen the existing data and might have important clinical implications because novel clonally unrelated lymphomas imitating relapses could possibly be treatable with less aggressive regimens compared to true recurrences
Open Access
Overexpression of vasoactive intestinal peptide receptors and cyclooxygenase-2 in human prostate cancer. Analysis of potential prognostic relevance
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Fernández-Martínez, Ana B.; Carmena, María J.; Arenas, María Isabel; Bajo, Ana M.; Prieto, Juan C.; Sánchez-Chapado, Manuel
Vasoactive intestinal peptide (VIP) is a potent inductor of cyclooxygenase-2 (COX-2) expression in human prostate cancer cell lines. There are conflicting data regarding the role of COX-2 in the progression of this disease. Here we examined the expression of VIP receptors (VPAC1 and VPAC2) and COX-2 in prostate cancer specimens. Correlations among protein levels and various clinicopathological factors and prognosis of patients were statistically analyzed. For these purposes, formaldehyde-fixed, paraffin-embedded prostate tissue specimens from 63 patients with prostate cancer and 9 control samples were used. The expression of VPAC1 and VPAC2 receptors and COX-2 was analyzed at mRNA levels by quantitative reverse transcriptase-PCR. The corresponding expression at protein level was studied by immunohistochemistry, scored as negative, weak, moderate, or strong, and correlated with different clinicopathological factors by means of multivariate analysis. 88% of prostate cancer tissues overexpressed VPAC1-receptor at mRNA level, 72% VPAC2-receptor and 77% COX-2. Simultaneous overexpression of the three genes was seen in 52% of patients. Similar overexpression patterns were observed at protein level. The correlation between VPAC1 and VPAC2 receptor protein levels was statistically significant. However, no significant correlations existed among protein levels of VPAC receptors and COX-2 with patient age, prostate-specific antigen (PSA) levels, tumor stage, Gleason score and survival time. The overexpression of VPAC1 and VPAC2 receptors and COX-2 in cancer tissue gives them a potential role as targets for diagnosis of prostate cancer but results do not support a clear value as biomarkers for the clinical prognosis of this disease.
Open Access
Effect of tempol on myocardial vascular remodeling in female spontaneously hypertensive rats
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Bonacasa Fernández, Bárbara; Hernández Albaladejo, Inmaculada; Fenoy Palacios, Francisco José; Quesada Pérez, Tomás; López Cano, Bernardo; Facultades, Departamentos, Servicios y Escuelas::Departamentos de la UMU::Fisiología
Objective: The present study evaluated whether the treatment with the superoxide anion dismutase mimetic tempol prevents the worsening in hypertension and in myocardial vascular remodeling induced by ovariectomy in female spontaneously hypertensive rats (SHR). Methods: Experiments were performed in ten week old female SHRs randomly assigned to the groups: intact (INT: given vehicle; INT+T: treated with tempol, 90 mg/kg/day), ovariectomized (OVX: vehicle and OVX+T: tempol, respectively) and ovariectomized treated with 17ß-estradiol (OVX+E2 and OVX+E2+T). Evolution of systolic blood pressure (SBP) was determined every other week in lightly restrained awake rats using a noninvasive computerized tail-cuff plethysmography system. At 18 weeks of age the heart was excised and structural changes in histopathological sections of coronary vessels were quantified on a computerized imaging system analyzer. Results: SBP was significantly lower in female SHRs treated with tempol compared to the values measured in untreated animals. In the vascular remodeling of myocardial arterioles, OVX+T rats had a lower media cross sectional area and media-to-lumen ratio than those observed in the OVX SHR. Interestingly, treatment with tempol in the presence of estradiol (in female INT and OVX+E2 SHR ) increased media cross sectional area and wall-to-lumen ratio of myocardial arterioles, despite the fact that it lowered arterial pressure in those groups. Conclusions: These results indicate that tempol prevents arterial hypertension and blunts myocardial vascular remodeling in ovariectomized SHR. Paradoxically, when tempol is given in presence of estradiol it has a detrimental effect on myocardial arteriolar remodeling
Open Access
Effects of Helicobacter pylori on biological characteristics of gastric epithelial cells
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Wang, Ping; Mei, Juan; Tao, Jing; Zhang, Ning; Tian, Hua; Fu, Guo-Hui
Infection with Helicobacter pylori (H. pylori) strains is linked to an increased risk of inflammation and gastric cancer. To investigate the effects of H. pylori on biological characteristics of gastric epithelial cells SGC-7901, derived from human adenocarcinoma, morphological appearances of both the pathogen and these cells, as well as features of attachment and internalization were observed by using transmission electron microscopy (TEM). We also investigated cell junctions and invasion by TEM and Transwell Invasion Assay. Cell proliferation and apoptosis were assessed by using chromogenic methylthiazol tetrazolium bromide (MTT) dye and flow cytometry. Three types of H. pylori were observed around, attaching to, or invading tumor cells. Cellular damage was characterized by vacuolar degeneration, dilated endoplasmic reticulum (ER), and reduction of organelles. Cell junctions and cell microvilli reduced or disappeared. H. pylori inhibited cell proliferation, whereas it had no effect on apoptosis. It also promoted gastric carcinoma cell invasion. H. pylori damages cell construction, destroys cell junctions, inhibits cell proliferation, promotes cell invasive ability, and, therefore, might accelerate the malignant progress and metastasis of gastric cancer
Open Access
Dendritic spines of the medial amygdala: plasticity, density, shape, and subcellular modulation by sex steroids
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Rasia-Filho, Alberto A.; Dalpian, Francine; Menezes, Itiana C.; Brusco, Janaína; Moreira, Jorge E.; Cohen, Rochelle S.
The medial nucleus of the amygdala (MeA) is a complex component of the “extended amygdala” in rats. Its posterodorsal subnucleus (MePD) has a remarkable expression of gonadal hormone receptors, is sexually dimorphic or affected by sex steroids, and modulates various social behaviors. Dendritic spines show remarkable changes relevant for synaptic strength and plasticity. Adult males have more spines than females, the density of dendritic spines changes in the course of hours to a few days and is lower in proestrous and estrous phases of the ovarian cycle, or is affected by both sex steroid withdrawal and hormonal replacement therapy in the MePD. Males also have more thin spines than mushroom-like or stubby/wide ones. The presence of dendritic fillopodia and axonal protusions in the MePD neuropil of adult animals reinforces the evidence for local plasticity. Estrogen affects synaptic and cellular growth and neuroprotection in the MeA by regulating the activity of the cyclic AMP response element-binding protein (CREB)-related gene products, brain-derived neurotrophic factor (BDNF), the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) and the activity-regulated cytoskeleton-related protein (Arc). These effects on signal transduction cascades can also lead to local protein synthesis and/or rearrangement of the cytoskeleton and subsequent numerical/morphological alterations in dendritic spines. Various working hypotheses are raised from these experimental data and reveal the MePD as a relevant region to study the effects of sex steroids in the rat brain
Open Access
Regular consumption of a silicic acid-rich water prevents aluminium-induced alterations of nitrergic neurons in mouse brain: histochemical and immunohistochemical studies
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Foglio, E.; Buffoli, B.; Exley, C.; Rezzani, R.; Rodella, L.F.
Silicon is not generally considered an essential nutrient for mammals and, to date, whether it has a biological role or beneficial effects in humans is not known. The results of a number of studies suggest that dietary silicon supplementation might have a protective effect both for limiting aluminium absorption across the gut and for the removal of systemic aluminium via the urine, hence, preventing potential accumulation of aluminium in the brain. Since our previous studies demonstrated that aluminium exposure reduces the number of nitrergic neurons, the aim of the present study was to compare the distribution and the morphology of NO-containing neurons in brain cortex of mice exposed to aluminium sulphate dissolved in silicic acid-rich or poor drinking water to assess the potential protective role of silicon against aluminium toxicity in the brain. NADPH-d histochemistry and nNOS immunohistochemistry showed that high concentrations of silicon in drinking water were able to minimize the impairment of the function of nitrergic neurons induced by aluminium administration. We found that silicon protected against aluminium-induced damage to the nitrergic system: in particular, we demonstrated that silicon maintains the number of nitrergic neurons and their expression of nitrergic enzymes at physiological levels, even after a 12 and 15 month exposure to aluminium
Open Access
Trehalose treatment accelerates the healing of UVB-irradiated corneas. Comparative immunohistochemical studies on corneal cryostat sections and corneal impression cytology
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Cejkova, Jitka; Cejka, Čestmír; Luyckx, Jacques
The UVB-irradiated cornea is damaged by oxidative stress. Toxic oxygen products induced by UVB radiation in the cornea are insufficiently removed by antioxidants, whose numbers decrease with increasing UVB irradiation. In addition, the UVB-irradiated cornea suffers from hypoxic conditions because damaged corneal cells cannot utilize oxygen normally, although the supply of oxygen to the cornea is unchanged (normal). This contributes to attenuated re-epithelialization, corneal neovascularization and apoptotic cell death. Our previous publications reported that trehalose applied on the corneal surface during irradiation significantly suppressed UVB-induced corneal oxidative damage. The results of this study provide for the first time important evidence that trehalose applied on the surface of corneas for two weeks following repeated UVB irradiation (312 nm, daily dose 0.5 J/cm2) accelerated corneal healing, restored corneal transparency and suppressed corneal neovascularization. Compared to buffered saline treatment, following which caspase-3, nitrotyrosine, malondialdehyde and urokinase-type plasminogen activator were still strongly expressed in the corneal epithelium two weeks after irradiation and corneal neovascularization was evident, apoptotic cell death was already significantly reduced after one week of trehalose application. The expression of other markers of injury returned to normal levels during two weeks of trehalose treatment. In conclusion, our results show that trehalose accelerated healing of the UVB irradiated cornea, very probably via suppression of hypoxia-response injury. In addition, immunohistochemical results on corneal cryostat sections corresponded with those obtained using corneal impression cytologies, thus confirming that corneal impression cytologies are useful for diagnostic purposes

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