Histology and histopathology Vol.31,nº 12 (2016)
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- PublicationOpen AccessNestin+cells forming spheroids aggregates resembling tumorspheres in experimental ENU-induced gliomas(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) García Blanco, Alvaro; Bulnes, Susana; Pomposo, Iñigo; Carrasco, Alex; Lafuente, José VicenteNestin+cells from spheroid aggregates display typical histopathological features compatible with cell stemness. Nestin and CD133+cells found in glioblastomas, distributed frequently around aberrant vessels, are considered as potential cancer stem cells. They are possible targets for antitumoral therapy because they lead the tumorigenesis, invasiveness and angiogenesis. However, little is known about their role and presence in low-grade gliomas. The aim of this work is to localize and characterize the distribution of these cells inside tumors during the development of experimental endogenous glioma. For this study, a single dose of Ethyl-nitrosourea was injected into pregnant rats. Double immunofluorescences were performed in order to identify stem-like and differentiated cells. Low-grade gliomas display Nestin+cells distributed throughout the tumor. More malignant gliomas show, in addition to that, a perivascular location with some Nestin+cells co-expressing CD133 or VEGF, and the intratumoral spheroid aggregates of Nestin/CD133+ cells. These structures are encapsulated by welldifferentiated VEGF/GFAP+cells. Spheroid aggregates increase in size in the most malignant stages. Spheroid aggregates have morphological and phenotypic similarities to in vitro neurospheres and could be an in vivo analogue of them. These arrangements could be a reservoir of undifferentiated cells formed to escape adverse microenvironments.
- PublicationOpen AccessEstrogen-mediated dental tissue regeneration(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Lu, Yadie; Jin, Lin; Lei, Gang; Fu, Yujin; Wang, Yanqiu; Yu, JinhuaAs the key regulator of hard tissue metabolism in both men and women, estrogen regulates the processes necessary for cell growth, proliferation, and differentiation through estrogen receptor (ER). Estrogen deficiency usually causes systemic osteoporosis not only in long bones but also in jaw bones, and exogenous estrogen can enhance the osteogenic potential of mesenchymal stem cells. Dental mesenchymal stem cells (DMSCs) represent a group of stem cells isolated from different parts of the tooth, including dental pulps, apical papillae and periodontal ligaments. A number of studies have proved that estrogen plays an important role in the proliferation, differentiation and tissue regeneration of human DMSCs. Thus, this review will focus on the effects of estrogen on proliferation, apoptosis, and differentiation of dental stem cells, discuss evidence from studies in rodents that estrogen plays an important role in dental morphogenesis as well as periodontal remodeling, and suggest directions for future studies in estrogen-related tooth regeneration.
- PublicationOpen AccessMixed acinar-neuroendocrine carcinoma of the pancreas: a case report and a review(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Jakobsen, Mark; Klöppel, Günter; Detlefsen, SönkeA 62-year-old woman presented with abdominal discomfort. Imaging studies showed a tumor in the pancreatic tail. At contrast-enhanced CT and macroscopy, the tumor showed cystic, solid and hemorrhagic areas. Histologically, the tumor was wellcircumscribed and entirely encapsulated. Some of the tumor cells in the cystic areas were reminiscent of acinar cells, and the majority was arranged in a solid growth pattern. Immunohistochemistry revealed >30% positivity for chymotrypsin, chromogranin A, synaptophysin, and CD56. The diagnosis of a mixed acinar-neuroendocrine carcinoma (MAEC) was made. Review of the English language literature revealed 44 previously published cases of resected MAECs. We found that, compared to pure acinar cell carcinoma, patients with MAEC have a slightly higher age and are less frequently males, as the male/female ratio was almost equal. The histogenesis of MAEC is still controversial. Due to the small number of cases it is at present not possible to define an evidencebased optimal treatment strategy for these patients.
- PublicationOpen AccessMuscular hypertrophy and atrophy in normal rats provoked by the administration of normal and denervated muscle extracts(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Agüera, Eduardo; Castilla, Salvador; Luque, Evelio; Jimena, Ignacio; Leiva Cepas, Fernando; Ruz Caracuel, Ignacio; Peña, JoséThis study was conducted to determine the effects of extracts obtained from both normal and denervated muscles on different muscle types. Wistar rats were used and were divided into a control group and four experimental groups. Each experimental group was treated intraperitoneally during 10 consecutive days with a different extract. These extracts were obtained from normal soleus muscle, denervated soleus, normal extensor digitorum longus, and denervated extensor digitorum longus. Following treatment, the soleus and extensor digitorum longus muscles were obtained for study under optic and transmission electron microscope; morphometric parameters and myogenic responses were also analyzed. The results demonstrated that the treatment with normal soleus muscle and denervated soleus muscle extracts provoked hypertrophy and increased myogenic activity. In contrast, treatment with extracts from the normal and denervated EDL had a different effect depending on the muscle analyzed. In the soleus muscle it provoked hypertrophy of type I fibers and increased myogenic activity, while in the extensor digitorum longus atrophy of the type II fibers was observed without changes in myogenic activity. This suggests that the muscular responses of atrophy and hypertrophy may depend on different factors related to the muscle type which could be related to innervation.
- PublicationOpen AccessHistological assessment for femora of ovariectomized obesity (db/db) mice carrying mutated leptin receptor(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Tanaka, Yusuke; Hasegawa, Tomoka; Yamada, Tamaki; Yamamoto, Tomomaya; Sasaki, Muneteru; Hongo, Hiromi; Tsuboi, Kanako; Haraguchi, Mai; Luiz de Freitas, Paulo Henrique; Li, Minqi; Oda, Kimimitsu; Totsuka, Yasunori; Tei, Kanchu; Amizuka, NorioIn order to provide a clue to understand the interplay between leptin and estrogen, we have examined femoral metaphyses of ovariectomized db/db mice carrying a mutated leptin receptor. We performed ovariectomy (OVX) or sham-operation (sham) on 12- week old female wild-type and db/db mice, and then, after 8 weeks, divided the animals into four groups: wild-type sham, wild-type OVX, db/db sham and db/db OVX. Samples from all groups were prepared for histochemical and ultrastructural examinations. As a result, db/db sham mice showed a reduced number and thickness of metaphyseal trabeculae and excessive adipose tissue when compared to wild-type sham mice. The wild-type OVX group exhibited markedly diminished trabecular number, as well as lower populations of osteoblasts and osteoclasts in comparison to wild-type sham group. On the other hand, trabecular numbers were similar for the two db/db groups, suggesting that the effect of the ovariectomy, i.e., estrogen deficiency may be lessened in this animal model. Leptin receptor was mainly found in osteoblasts and in bone marrow stromal cells including adipocytes. In addition, the expression of estrogen receptor did not seem to change after OVX in wild-type mice and in db/db mice. Both db/db sham and OVX mice featured many adipocytes close to the metaphyseal chondroosseous junction, while osteoblasts accumulated glycogen granules and lipid droplets. Therefore, it seems likely that the disruption of leptin signaling in db/db mice shifts the cell differentiation cascade towards the adipocyte lineage, resulting in an osteoporotic bone independently of estrogen deficiency.
- PublicationOpen AccessTelocytes and lung disease(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Song, Dongli; Cretoiu, Dragos; Cretoiu, Sanda M.; Wang, XiangdongTelocytes (TCs) represent a new distinct type of interstitial cells found in many organs, including lungs. TCs are mainly defined by a small cellular body from which arise very long (hundreds of micrometers) extensions named telopodes. During the last years, TCs were characterized in respect with their microRNA profiles, gene features and proteome signatures. Also, the ultrastructural 3D configuration was further elucidated by the aid of the FIB-SEM technology. TCs are able to communicate by homo- and heterocellular contacts with neighboring cells and are also able to transfer genetic information and signaling molecules to influence other cells by means of extracellular vesicle release. However, the exact function of lung TCs remains unclear. Here, we review the potential significance of TCs in the pathogenesis of pulmonary diseases. We will also discuss some future possibilities for targeting TCs as a potential therapeutic strategy.
- PublicationOpen AccessMorphologic changes within the cerebellar cortex in the unilateral 6-hydroxydopamine lesioned rat model for Parkinson disease(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Wu, Chenghua; Fan, Guoguang; Wu, Chunli; Yu, Guibo; Li, ZixuanParkinson’s disease (PD) is a common neurodegenerative disorder caused by the progressive loss of dopaminergic neurons in the substantia nigra. Most investigations have focused on the cerebral regions such as the basal ganglia, thalamus, or the substantia nigra, but whether there is pathologic impairment within the cerebellum has rarely been assessed. Synapsin and neurofilament as the inner markers of neurons and synapses reflect the functional state by their distribution or expression. Significant morphologic changes at the cellular level have been demonstrated directly or indirectly in multiple neurodegenerative diseases. The purpose of this study was to determine whether the behavioral abnormalities that accompany PD are associated with the cerebellum using an in vivo 6- hydroxydopamine lesioned rat model. Forty-two rats were divided into three groups, the Parkinsonian group (N=22), sham group (N=10) and control group (N=10). The dopaminergic lesion was determined by immunohistochemical analysis for tyrosine hydroxylaseimmunopositive cells. Immunohistochemical studies showed that the density of synapsin I in the granular layer of the cerebellum on both sides of the Parkinsonian -model was not statistically significantly different compared to the control and sham groups. However, expression of neurofilament H in the cortex within bilateral paramedian lobule (PML) and Crus 2 of the ansiform lobule (C2AL) in cerebellum posterior lobe of Parkinsonian rats was decreased compared with controls (P<0.05), especially in the loss of Purkinje cells and the presence of morphologic abnormalities in the cell nucleus. The study suggested that loss of neurons and synapses may take place in the cerebellar cortex of Parkinson’s disease, and might play an important role in the pathologic mechanism of PD.
- PublicationOpen AccessCancer stem cell as therapeutic target for melanoma treatment(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Alamodi, Abdulhadi A.; Eshaq, Abdulaziz M.; Hassan, Sofie Yasmin; Hmada, Youssef Al; El Jamal, Siraj M.; Fothan, Ahmed M.; Arain, Omair M.; Hassan, Sarah-Lilly; Haikel, Youssef; Megahed, Mosaad; Hassan, MohamedHuman malignant melanoma is a highly aggressive skin tumor that is characterized by its extraordinary heterogeneity, propensity for dissemination to distant organs and resistance to cytotoxic agents. Although chemo- and immune-based therapies have been evaluated in clinical trials, most of these therapeutics do not show significant benefit for patients with advanced disease. Treatment failure in melanoma patients is attributed mainly to the development of tumor heterogeneity resulting from the formation of genetically divergent subpopulations. These subpopulations are composed of cancer stem-like cells (CSCs) as a small fraction and non-cancer stem cells that form the majority of the tumor mass. In recent years, CSCs gained more attention and suggested as valuable experimental model system for tumor study. In melanoma, intratumoral heterogeneity, progression and drug resistance result from the unique characteristics of melanoma stem cells (MSCs). These MSCs are characterized by their distinct protein signature and tumor growth-driving pathways, whose activation is mediated by driver mutation-dependent signal. The molecular features of MSCs are either in a causal or consequential relationship to melanoma progression, drug resistance and relapse. Here, we review the current scientific evidence that supports CSC hypothesis and the validity of MSCs-dependent pathways and their key molecules as potential therapeutic target for melanoma treatment.
- PublicationOpen AccessAnalysis of HER2 status in gastroesophageal tumor specimens using a new automated HER2 IQFISH pharmDx™ (Dako Omnis) assay(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Viale, Giuseppe; Paterson, Jennifer; Bloch, Miriam; Csathy, George; Allen, David; Dell’Orto, Patrizia; Kjærsgaard, Gitte; Levy, Yaron Y.; Trøst Jørgensen, JanThe human epidermal growth factor receptor 2 (HER2) is an important target for treatment of gastroesophageal cancer. Different slide-based assays are available for assessment of HER2 status. Overexpression of the HER2 protein is assessed by immunohistochemistry (IHC) whereas amplification of the HER2 gene is assessed by fluorescence in situ hybridization (FISH) or other in situ hybridization (ISH) methods. Here we report a summary of the validation data on HER2 IQFISH pharmDx™ (Dako Omnis), a newly developed assay for the automated staining platform Dako Omnis. This assay uses a non-toxic buffer that significantly reduces the hybridization time, which results in a total turnaround time of less than 4 hours from deparaffinization to counting of the gene and centromere signals. The data reported in the current summary cover method comparison, assessment of staining quality, observer-to-observer reproducibility as well as reproducibility within and between laboratories. Based on data from the different studies it was concluded that HER2 IQFISH pharmDx (Dako Omnis) is a reliable and robust assay, with high precision and at least comparable to the manual HER2 IQFISH pharmDx™ assay. The HER2 IQFISH pharmDx (Dako Omnis) assay is currently not commercially available outside the European Union.