Histology and histopathology Vol.33, nº9 (2018)
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- PublicationOpen AccessThrombocytopenia in leukemia: Pathogenesis and prognosis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Shahrabi, Saeid; Maleki Behzad, Masumeh; Jaseb, Kaveh; Saki, NajmaldinLeukemias, a heterogeneous group of hematological disorders, are characterized by ineffective hematopoiesis and morphologic abnormalities of hematopoietic cells. Thrombocytopenia is a common problem among leukemia types that can lead to hemorrhagic complications in patients. The purpose of this review article is to identify the conditions associated with the incidence of thrombocytopenia in leukemias. It can be stated that although translocations have been considered responsible for this complication in many studies, other factors such as bone marrow failure, genes polymorphism, a mutation in some transcription factors, and the adverse effects of treatment could be associated with pathogenesis and poor prognosis of thrombocytopenia in leukemias. Considering the importance of thrombocytopenia in leukemias, it is hoped that the recognition of risk factors increasing the incidence of this complication in leukemic patients would be useful for prevention and treatment of this disorder.
- PublicationOpen AccessUsefulness of identifying G-protein-coupled receptor dimers for diagnosis and therapy of neurodegenerative diseases and of gliomas(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Reyes Resina, Irene; Aguinaga, David; Labandeira García, José Luis; Lanciego, José Luis; Navarro, Gemma; Franco, Rafaely. Immunochemical detection of G-proteincoupled receptors (GPCRs) in cells and tissues was a technical challenge for years. After the discovery of formation of GPCR dimers/trimers/tetramers in transfected cells, a most recent challenge has been to confirm receptor-receptor interactions in natural sources. The occurrence of dimers or higher order oligomers is important from a therapeutic point of view, mainly because their physiology/pharmacology is different from those of individual receptors. On the one hand, pathophysiological factors need to count more on GPCR dimers than on individual receptors. On the other hand, the expression of dimers, trimers, etc. may change in pathological conditions and/or along the course of a disease. This review will focus on G-protein-coupled receptor dimers, on how to detect them by novel histological techniques and on how the detection may be used in diagnosis and therapy of ailments of the central nervous system, for instance in neurodegenerative diseases and gliomas.
- PublicationOpen AccessHuman skin model for mimic dermal studies in pathology with a clinical implication in pressure ulcers(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Cristóbal, Lara; Ortega, Miguel A.; Asúnsolo, Ángel; Romero, Beatriz; Álvarez Mon, Melchor; Buján, Julia; Maldonado, Andrés A.; García Honduvilla, NatalioDespite advances in regenerative medicine and tissue engineering, human skin substitutes remain a clear goal to achieve. Autografts remain the principal clinical option. The long-term changes in dermis, as well as its response after injuries, are not well known. Research in this field has been hindered by a lack of experimental animal models. This study analyzes the architectural dermal scaffold (collagen and elastin fibers plus fibrillin-microfibrils) changes in a model of human skin pressure ulcers in mice. Immunosuppressed NOD/Scid mice (n=10) were engrafted with human skin of dimensions 4x3 cm. After 60 days as a permanent graft, a pressure ulcer (PU) was created in the human skin using a compression device. Three study groups were established: full-thickness skin graft before (hFTSG) and after applying mechanical pressure (hFTSG-PU). Native human skin was used as control group. Evaluations were conducted with visual and histological assessment. Scaffold components from each group were compared by immunohistochemical staining (tropoelastin, collagen I and III, metalloproteins (MMP), fibulins, and lysil oxidases (LOX) among others). The long-term engrafted skin showed a certain degradative state of dermis scaffold, as noticed by the active expression of MMPs and tropoelastin compared to native skin. However, a great reparative response after pressure ulcer onto the engrafted skin was observed. A significant increase of fibrillin microfibrils components (TGF-β, MAGP-1 and fibrillin-1), and matrix suprastructures of collagen I, III and LOX lead to an active restructuration of dermal tissue. Our human skin model in mice revealed the important role of the dermal scaffold component to reach skin stability and its capability to react to mechanical pressure injuries. These results showed the important role of dermal scaffold to support the histoarchitecture and mechanosensation of the human skin.
- PublicationOpen AccessUnderstanding the roles of negative immune regulator TIPE2 in different diseases and tumourigenesis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Li, Zequn; Jia, Wenyu; Niu, Jun; Zhang, LiningTumour necrosis factor (TNF)-alpha-induced protein 8-like 2 (TIPE2 or TNFAIP8L2) is a novel identified negative regulator of both innate and adaptive immunity, which participates in immune homeostasis. TIPE2 is expressed in both immune tissues and nonimmune tissues. It is significantly down-regulated in patients with infectious and autoimmune diseases, and the depletion of TIPE2 causes severe inflammatory disease. In recent years, accumulating evidence has shown that TIPE2 serves as an ideal biomarker and as a tumour suppressor in several tumour types. In this review, we summarized the roles and corresponding mechanisms of TIPE2 in immune regulation and different diseases, especially in tumourigenesis.
- PublicationOpen AccessCircular RNAs: Characteristics, function and role in human cancer(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Chen, Shuo; Zhao, YangCircular RNAs (circRNAs) are universal and diverse endogenous noncoding RNAs (ncRNAs) that are widely found in eukaryotic cells. They form a covalently closed, continuous stable loop structure without 3' or 5' tails, which are different from linear RNAs. As new members of the ncRNAs, circRNAs’ formation, function, and mechanism are attracting increased research attention. CircRNAs play important roles in all kinds of cancer and may be potential novel biomarkers and therapeutic targets for cancer treatment through their function as microRNA (miRNA) molecular “sponges”, RNA-binding protein (RBP) sponges, protein translators, and gene transcription regulators. In this review, we introduce the formation and function of circRNAs, and summarize the biological effects of circRNA in tumorigenesis and progression, providing evidence for the potential use of circRNAs in the diagnosis and clinical treatment of cancer.
- PublicationOpen AccessExpression of plakophilin 3 in diffuse malignant pleural mesothelioma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Mašić, Silvija; Brčić, Luka; Krušlin, Božo; Pigac, Biserka; Stančić Rokotov, Dinko; Jakopović, Marko; Seiwerth, SvenDiffuse malignant pleural mesothelioma (DMPM) is the most common primary malignant pleural neoplasm still posing major diagnostic, prognostic and therapeutic challenges. Plakophilins are structural proteins considered to be important for cell stability and adhesion in both tumor and normal tissues. Plakophilin 3 is a protein present in desmosomes of stratified and simple epithelia of normal tissues with presence in malignant cells of various tumors where it participates in the process of tumorigenesis. The aim of this study was to investigate the expression of plakophilin 3 protein in DMPM, but also to study its prognostic significance and relation to histologically accessible parameters of aggressive growth. Archival samples of tissue with established diagnosis of DMPM and samples of normal pleural tissue were used. Tumor samples were classified into three histological types of DMPM (epithelioid, sarcomatoid and biphasic). Additional subclassification of epithelioid mesotheliomas into nine patterns based on the prevalent histological component of the tumor was then performed. After immunohistochemical staining, cytoplasmic and membrane immunopositivity of tumor cells was assesed by scoring the intensity of the staining from 0 (no staining) to 4 (very strong staining). Prognostic value and expression of plakophilin 3 with consideration to histologically estimated aggression in tumor growth were then statistically analyzed using nonparametric tests. The results demonstrated higher level of plakophilin 3 expression in tumor samples with histologically more aggressive tumor growth, but no significant prognostic value. According to our study, plakophilin 3 appears to be involved in tumor invasion in malignant mesothelioma.
- PublicationOpen AccessExpression of EMT inducers integrin-linked kinase (ILK) and ZEB1 in phyllodes breast tumors is associated with aggressive phenotype(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Akrida, Ioanna; Nikou, Sofia; Gyftopoulos, Konstantinos; Argentou, Marianna; Kounelis, Sofia; Zolota, Vassiliki; Bravou, Vasiliki; Papadaki, HelenPhyllodes tumors (PTs) of the breast constitute an uncommon group of mammary fibroepithelial lesions with ambiguous biologic behavior. Recent evidence suggests that epithelial mesenchymal transition (EMT), a driving force of cancer progression is implicated in PTs pathogenesis. Integrin-linked kinase (ILK), a focal adhesion kinase, has been implicated in cancer and EMT and represents a novel cancer therapeutic target. In this study, we aimed to investigate ILK and EMT markers expression in phyllodes breast tumors in relation to tumor grade. Expression of ILK and EMT markers E-cadherin, β-catenin, Ν-cadherin, vimentin, Snail, ZEB1 and Twist was evaluated by immunohistochemistry in paraffin-embedded tissue sections from 96 human phyllodes breast tumors (48 benign, 27 borderline, 21 malignant). Cytoplasmic and nuclear immunopositivity of ILK were observed in both the epithelial and the stromal component of phyllodes breast tumors and were significantly higher with increasing tumor grade. An EMT-related expression profile consisting of decreased membranous and increased nuclear/cytoplasmic immunoreactivity of Ecadherin and β-catenin and increased expression of Ncadherin, vimentin, Snail, ZEB1 and Twist was observed in tumor epithelial and stromal component and was significantly associated with malignant phyllodes breast tumor histopathology. Interestingly, there was a significant correlation of ILK expression with all of the EMT markers examined. Our results suggest that EMT significantly contributes to phyllodes tumor pathogenesis and originally implicate ILK and ZEB1 in phyllodes tumors malignant phenotype.
- PublicationOpen AccessSPHK2 protein expression, Ki-67 index and infiltration of tumor-associated macrophages (TAMs) in human glioma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Liu, J.; Zhou, Q.; Wu, C.P.; Xu, Y.W.; Liu, W.L.; Zhao, H.F.; Li, W.P.Introduction. Sphingosine kinases (SPHKs), the Ki-67 index and tumor-associated macrophages (TAMs) are associated with diverse human malignancies, including glioma. SPHK2, a subtype of SPHKs, has not been assessed in glioma or correlated with the Ki-67 index or TAM infiltration. We tested the hypothesis that expression of SPHK2 correlates with the Ki-67 index and TAM infiltration in patients with glioma. Materials and method. Western blot analysis was performed on protein lysates prepared from human astrocyte (HA) and glioma cell lines. Immunofluorescence was used to determine the subcellular location of SPHK2 protein in glioma cells. Next, immunohistochemistry was employed to analyze the correlations among SPHK2, Ki-67, CD68, and CD206 in 11 non-neoplastic brain tissues and 60 glioma tissues. All slides were evaluated under ×400 magnification, and the ratio of positively stained cells to the total number of cells was calculated for analysis. Results. SPHK2, CD68 and CD206 were all increased in glioma tissues compared to non-neoplastic brain tissues, but there were no differences between WHO grades of glioma. Ki-67 was highest in WHO grade IV tumors and lowest in non-neoplastic brain tissues, and all between-group differences were statistically significant. Moreover, SPHK2 expression was positively correlated with the Ki-67, CD68 and CD206 indexes. Finally, the CD68 and CD206 indexes were both associated with the Ki-67 index. Conclusion. SPHK2 protein expression, the Ki-67 index and TAM infiltration in human glioma tissue were reported in this study for the first time. SPHK2 was positively associated with TAM infiltration and glioma proliferation. Mechanistically, SPHK2 may promote glioma growth by stimulating TAMs to polarize M2-type macrophages.
- PublicationOpen AccessLamellation in fibrous dysplasia: a clinicopathologic study(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Fuat Cicek, Ali; Kilinc, Melih; Safali, Mukerrem; Gunhan, OmerBackground. Fibrous dysplasia (FD) is a maturation defect characterized by immature woven bones and stroma. However, especially in craniofacial bones, lamellation can be seen and this is associated with the maturation. Aim: To show maturation in FD and discuss the factors that may affect the maturation. Materials and Methods. Ninety-five FD cases were divided into three subgroups according to the lamellation percentage as Groups 1, 2 and 3 (low, moderate and high lamellation, respectively). Each group was compared in terms of the peritrabecular clefting (PTC), stromal cellularity and the age. The lesions under pressure and the ones that are not were compared in terms of lamellation percentage. Results. A significant statistical difference was found between Groups 1 and 3 in terms of PTC, stromal cellularity, histologic pattern suggesting maturation (p<0.001, p<0.001, p=0.002, respectively). Conclusion. The findings suggested a strong relation between lamellation and maturation. Lamellation was more prominent in the bones under pressure than the others. Considering lamellation as a finding of maturation, it is possible to establish a relation between maturation and pressure. Therefore, future studies should focus on the question if the pressure could be a factor for maturation and it could be used for treating FD.
- PublicationOpen AccessMatrix metalloproteinase-1 (MMP-1) and (MMP-8) gene polymorphisms promote increase and remodeling of the collagen III and V in posterior tibial tendinopathy(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Diniz Fernandes, Tulio; Godoy Santos, Alexandre Leme; Santos, Maria Cristina; Pontin, Pedro; Alves Pereira, Caio Augusto; Justi Jardim, Yuri; Pereira Velosa, Ana Paula; Maffulli, Nicola; Teodoro, Walcy Rosolia; Capelozzi, Vera LuizaPosterior tibial tendinopathy (PTT) can lead to acquired flatfoot in adults. Many patients develop PTT without any identifiable risk factors. Molecular changes in extracellular matrix (ECM) and matrix metalloproteinase (MMP) polymorphism may influence the risk of developing PTT. We aim to investigate the association between matrix metalloproteinase-1 (MMP1) and (MMP-8) gene polymorphisms with changes in collagen I, III and V in PTT. A case-control study with 22 patients and 5 controls was performed. The MMP-1 (2G/2G) and MMP-8 (T/T) genotypes were determined by PCR-restriction fragment length polymorphism. Tendon specimens were evaluated by a histologic semiquantitative score, immunofluorescence and histomorphometry for collagen I, III and V. Tendon specimens from PTT demonstrated marked distortion of the architecture with necrosis, large basophilic areas with disruption of the normal linear orientation of collagen bundles, infiltration of inflammatory cells, dystrophic calcification and ossification. Under immunofluorescence, PTT tendon specimens showed weak green fluorescence and diffuse distribution of collagen I fibers, but strong fluorescence of collagen III and V. The collagen I fibers were significantly decreased whereas an increase of collagen III and V were found in PTT compared to control groups. In addition, PTT group presented a significant association with MMP-1 and MMP-8 gene polymorphisms. Patients with PTT matrix metalloproteinase-1 (MMP-1) and (MMP-8) gene polymorphisms presented an increase of the collagen III and V ratio, suggesting that the higher proportion in degenerated tendons could contribute to a decrease in the mechanical resistance of the tissue. Still, functional and association studies are needed to elucidate evident roles of MMPs in PTT.
- PublicationOpen AccessIn vivo toxicity of the ribosome- inactivating lectin ebulin f in elderly mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Garrosa, Manuel; Jiménez, Pilar; Córdoba Díaz, Damián; García Recio, Verónica; Gayoso, Sara; Rojo, María Ángeles; Gayoso, Manuel J.; Girbés, TomásEbulin f is a ribosome-inactivating protein (RIP) present in green fruits of the dwarf elder (Sambucus ebulus L). Since dwarf elder fruits are used for food and as a medicine, we assessed the study of toxicological effects and safety of ebulin f in elderly mice, comparing these results with those reported in young animals and with other RIPs. Female Swiss mice aged 6 and 12 months of age were intraperitoneally injected with a single dose from 1.4 to 4.5 mg/kg ebulin f. Heart, stomach, intestines, lung, kidney, liver, spleen, pancreas, adrenal gland, uterus, ovary and brain were studied. Histology analysis was carried out by staining with hematoxylin and eosin and Masson’s trichrome observed with a light microscope, or apoptosis detection by TUNEL method observed with a confocal laser microscope. Treated animals injected with the lower dose could recover their weights, but after 14 days half of them died. The higher dose caused a progressive loss of body weight leading to death. In the animals of the experimental groups it was found atrophy of Lieberkühn’s crypts, pneumonia, nephronal degeneration, myocardial atrophy, centrolobular hepatic necrosis, splenic white pulp necrosis foci and increased rate of apoptosis in the intestines and liver, in which apoptoses were mainly located in the vicinity of the lobular central vein. We conclude that ebulin f affects vital organs in elderly mice.
- PublicationOpen AccessIdentification of serotoninergic system components in stallion sperm(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Jiménez Trejo, F.; Coronado Mares, I.; Boeta, M.; González Santoyo, I.; Vigueras Villaseñor, R.; Arriaga Canon, C.; Herrera, L.A.; Tapia Rodríguez, M.Assisted reproductive techniques have been used on several domestic animals to preserve desirable traits in strains of high genetic and commercial value; however in equines its efficiency rate is relatively low. To increase the conception ratio in stallions, some research groups have used pharmacological treatments which promote sperm hyperactivation in order to increase male’s fertility rates. In this way, our previous work suggests that serotonin (5-HT) could be a good pharmacological candidate that facilitates conception rate in domestic horses. 5-HT is a neurohormone involved in several reproductive processes, i.e., it enhances hyperactivation, motility, and promotes the acrosome reaction in mammalian sperm, but it has not been described in the stallion sperm yet. Therefore, using both immunofluorescence and western blot techniques, we searched for and found some serotonin markers such as 5-HT, 5-HT1B, 5-HT2A, 5-HT3 receptors, both TPH1 and MAOA enzymes, and serotonin transporter (5-HTT) in stallion sperm. In addition, we found a non-neuroendocrine cell, V-MAT1 transporter, which has not been previously reported in mammalian sperm. Our results suggest that serotoninergic system is present in stallion sperm, which could be a pharmacological target to increase the conception rates in domestic horses.