Browsing by Subject "Immunohistochemical"
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- PublicationOpen AccessAntigenic profile of human bronchial gland(Murcia : F. Hernández, 2005) Sánchez-Mora, N.; Rendón-Henao, J.; Monroy, V.; Herranz Aladro, M.; Álvarez-Fernández, EmilioBronchial glands have been regarded as modified salivary glands. It is well known that there no previous reviews concerning the antigenic profile of the bronchial wall. The aim of this study is a systematic survey of the antigenic profile and to describe the histology of normal human bronchial glands. Six formalin-fixed, paraffin-embedded surgical specimens were studied using a panel of 22 polyclonal and monoclonal antibodies by the avidin-biotin-peroxidase method. Bronchial glands disclosed a tubuloacinar structure. The smallest ducts intercalated originated from a cluster of secretory acini and converge to form an excretory duct. No striated duct was observed. Acinar united is composed by mucous, serous and mixed units. Myoepithelial cells are found in relation to the intercalated ducts and secretory acinis. Secretory cells of bronchial glands reacted strongly with cytokeratin AE1 and moderately for CK7, CK18. Additionally, serous acinar cells reacted with AE3, CK19, CK5/6/8/18, CK8/18/19, and Leu7. Myoepithelial cells reacted strongly with a-smooth muscle actin, CD10 and CK34ßE12. Ductal system cells differed from acinar secretory cells in expressing CK34ßE12 and HSP27. In conclusion, the detailed knowledge of the immunohistochemical reactivities of normal cell types of normal human bronchial glands will prove useful in studies of bronchial pathology, especially of neoplastic processes.
- PublicationOpen AccessClinicopathological variables, immunophenotype, chromosome 1p36 loss and tumour recurrence of 247 meningiomas grade I and II(Murcia : F. Hernández, 2010) Ruiz, Juan; Martínez, Armando; Hernández, Susana; Zimman, Horacio; Ferrer, Milagros; Fernández, Cristina; Sáez, Mamen; López-Asenjo, José A.; Sanz Ortega, JuliánThe WHO grading scheme distinguishesbenign (grade I), atypical (grade II) and anaplastic(grade III) meningiomas. Both atypical and anaplasticmeningiomas exhibited an overall increased rate ofrecurrence, but between 15-20% benign meningiomaswill also exhibit an unfavourable clinical course withrecurrence before 10 years despite aggressive surgery.We investigated 247 cases of meningiomas grade I andII. The immunohistochemical expression of 30 differentmolecular biomarkers of cell adhesion molecules, cell-cycle and apoptosis regulators and checkpoints wasanalyzed. We also determined apoptosis by in-situhybridization (APOPDETEK™) and loss ofchromosome 1p36 by FISH. The study revealed astatistically significant co-variation (p<0.05) betweenmeningiomas grade II associated with severalclinicopathological features (Simpson grade of clinicalresection, necrosis, nuclear atypia, macronucleoli,transition to small cell, sheet-like growth, highcellularity), increased expression of several biomarkersof tumour proliferation (Cyclin A, Cyclin E, MIB-1 orMDM2), proteases (Cathepsin D) or cell-adhesion(CD44) and lower expression of progesterone receptorsthan meningiomas grade I. The presence of Psammomabodies or the location at convexity were protectiveprognostic factors for tumour recurrence while highcellularity and early age of onset (<57 year-old) wereindicators of increased recurrence risk. The expressionof COX-2, γ-catenin, Topoisomerase IIa, VEGF andMIB-1 was significantly higher in the cohort of recurrentmeningiomas. Meningiomas with chromosome 1p36 lossshowed a higher recurrence rate (33.3%) than meningiomas with normal chromosome 1p36 (18%).Increased COX-2 expression in recurrent meningiomamay also suggest a putative role of COX-2 inhibitors asa chemopreventive treatment for recurrence.
- PublicationOpen AccessExpression of smoothelin and smooth muscle actin in the skin(Editores F. Hernandez y Juan F. Madrid. Murcia, Universidad de Murcia, Departamento de Biologia Celular e Histologia, 2011) Aneiros Fernández, José; Husein-ElAhmed, Husein; Arias-Santiago, Salvador; Campos, Antonio; Carriel, Víctor; Sánchez-Montesinos, Indalecio; Garcia del Moral, Raimundo; Sánchez, Guillermo; O’Valle, Francisco; Aneiros, J.Introduction: Smoothelin is a cytoskeletal protein of differentiated smooth muscle cells with contractile capacity, distinguishing it from other smooth muscle proteins, such as smooth muscle actin (SMA). Objective: To evaluate the expression of smoothelin and SMA in the skin in order to establish specific localizations of smoothelin in smooth muscle cells with high contractile capacity and in the epithelial component of cutaneous adnexal structures. Methods: Immunohistochemical analysis (smoothelin and SMA) was performed in 18 patients with normal skin. Results: SMA was expressed by the vascular structures of superficial, deep, intermediate and adventitial plexuses, whereas smoothelin was specifically expressed in the cytoplasm of smooth muscle cells of the deepest vascular plexus and in no other plexus of the dermis. The hair erector muscle showed intense expression of smoothelin and SMA. Cells with nuclear expression of smoothelin and cytoplasmic expression of SMA were observed in the outer root sheath of the inferior portion of the hair follicles and intense cytoplasmic expression in cells of the dermal sheath to SMA. Conclusions: We report the first study of smoothelin expression in normal skin, which differentiates the superficial vascular plexus from the deep. The deep plexus comprises vessels with high contractile capacity, which is important for understanding dermal hemodynamics in normal skin and pathological processes. We suggest that the function of smoothelin in the outer root sheath may be to enhance the function of SMA, which has been related to mechanical stress.
- PublicationOpen AccessHeat shock proteins and survivin, Relationship and effects on proliferation index of retinoblastoma cells(Murcia : F. Hernández, 2008) Jiang, L.-B.; Liu, X.-Q.; Li, B.; He, X.-J.; Jin, Y.L.; Li, L.-Q.; Gao, F.; Wang, N.-L.Survivin and HSPs (heat shock proteins) are important anti-apoptotic proteins. However, limited research has been done regarding the collective effects of HSPs and survivin on the proliferative activities of RB cells. The purpose of this study was to narrow this gap by focusing on the expression of HSP70 and HSP90 and the interaction of these proteins with survivin. The proliferative activities of RB cells were analyzed by assessing the Ki-67 labeling index. Ki-67 recognizes a nuclear antigen expressed in all phases of the cell cycle except G(0) and early G(1), which makes it an excellent marker of cells in the proliferative phase. Immunohistochemical procedures were performed on retinal tissues from 43 RB patients who had undergone enucleation. Expression of HSP70, HSP90 and survivin was found in 65.12%, 86.05% and 62.79% of the cases respectively. No expression of any of these markers was found in normal retinal tissues. Expression of survivin was more frequent when HSP90 was detected than when HSP90 was not detected (P<0.05). The Ki-67 labeling index was higher in cases in which HSP90 or survivin was found than in cases in which neither protein was found (P<0.05). The Ki-67 labeling index was higher in cases positive for both HSP90 and survivin than in cases in which neither protein or only one protein was found (P<0.05). Expression of HSP70 neither correlated with that of survivin, nor had any significant effect on the Ki- 67 labeling index (P>0.05). Although expression of HSPs and survivin and the Ki-67 labeling index did not correlate with histopathologic typing of RB (P>0.05), our findings demonstrate that expression of HSP90 correlates with that of survivin in RB and the coexistence of survivin and HSP90 probably plays an important role in cellular proliferation in RB. Further work is indicated to clarify the role of these processes in progression of RB.
- PublicationOpen AccessImmunohistochemical localization of renin, NO synthase-1, and cyclooxygenase-2 in rodent kidney(Murcia : F. Hernández, 2008) Ichii, Osamu; Yabuki, Akira; Ojima, ToshimichiThe renin-angiotensin system (RAS) and tubuloglomerular feedback (TGF) are central to the maintenance of blood pressure and body fluid composition. Renin, NO synthase-1 (NOS-1), and cyclooxygenase-2 (COX-2) are key regulators of the RAS and TGF. In the present study, to investigate species-specific differences in the RAS and TGF, we immunohistochemically and morphometrically investigated the localization of renin, NOS-1, and COX- 2 in the kidneys of various laboratory rodents and comparing males with females (DBA/2Cr mice, F344/N rats, Syrian hamsters, MON/JmsGbs gerbils and Hartley guinea pigs). In all animals, renin-positive immunoreactions were observed in the vascular walls of afferent arterioles. Renin immunoreactions appeared to be more widely distributed in mice. Mice had a greater number of renin-positive arterioles than other species. NOS-1-positive reactions were detected in the macula densa (MD) of all animals. Mice had the greatest number of NOS-1-positive MD cells. In addition to NOS-1- positive reactions, COX-2-positive reactions were observed in the MD of mice, rats, hamsters and gerbils. Interestingly, guinea pigs had no COX-2-positive MD cells. Rats had the greatest number of COX-2-positive MD cells. In nephron segments excluding the MD, the immunohistochemical localization of NOS-1 and COX-2 differed markedly among not only species but also sexes within the same species. In conclusion, we determined that localization of renin, NOS-1, and COX-2 showed large species- and sex-related differences. These data suggest that the regulation mechanisms of the RAS and TGF via renin, NOS-1, and COX-2 differ among rodents.
- PublicationOpen AccessMucoepidermoid tumors of the bronchus. Ultrastructural and immunohistochemical study. Histiogenic correlations(Murcia : F. Hernández, 2007) Sánchez-Mora, N.; Parra-Blanco, V.; Cebollero-Presmanes, M.; Carretero-Albiñana, L.; Herranz, M.L.; Álvarez-Fernández, E.Bronchial mucoepidermoid tumors are uncommon neoplasms, morphologically similar to their salivary gland counterpart. The histogenesis is controversial. The aim of this study is to identify myoepithelial cells and speculate on their role in the origin of these tumors. Methods and Results: Sixteen bronchial mucoepidermoid tumor surgical specimens were formalin-fixed, paraffin-embedded and studied using a panel of nine antibodies in order to identify a myoepithelial differentiation. Additional antigens against several cytokeratins were performed in four cases and five of the biopies were studied using the electron microscopy. The different types of cells of the primary bronchial mucoepidermoid tumor (mucous luminal, intermediate and squamous) reacted strongly against AE1, CK7, 34bE12 and weakly with AE3, CK18 and CK8/18/19. S-100, a-smooth muscle actin, muscle actin HHF35 and a-actinin were consistently negative in all cell types. CD10 was positive in very few cells in just one case. Conclusion: The immunohistochemical and the ultrastructural study of bronchial mucoepidermiod tumors support a ductal unit origin, without a myoepithelial participation.
- PublicationOpen AccessP-cadherin, Vimentin and CK14 for identification of basal-like phenotype in breast carcinomas: an immunohistochemical study(Murcia : F. Hernández, 2010) Sousa, Bárbara; Paredes, Joana; Milanezi, Fernanda; Lopes, Nair; Martins, Diana; Dufloth, Rozany; Vieira, Daniella; Albergaria, André; Veronese, Luiz; Carneiro, Vitor; Carvalho, Silvia; Costa, José Luis; Zeferino, Luiz; Schmitt, FernandoIntroduction: The most suitable immunohistochemical criterion to identify basal-like breast carcinomas (BLBC), a molecular subgroup of breast cancer associated with poor prognosis, is the triple negative phenotype along with CK5 and/or EGFR immunoreactivity. However, several putative basal markers have been suggested as alternatives to identify BLBC with more accuracy. Experimental Design: The expression of CK5, EGFR, P-cadherin, CK14, Vimentin and p63 were evaluated in 462 invasive breast carcinomas to determine their sensitivity and specificity for BLBC identification. Results: P-cadherin and CK5 showed higher sensitivity values, while EGFR, Vimentin and CK14 were the most specific markers. The combination of CK5 with P-cadherin, Vimentin or CK14 proved to be a reliable option for distinguishing the basal phenotype, compared to the “gold standard” pair CK5/EGFR. Furthermore, P-cadherin was still able to recognize a large number of putative BLBC among the “unclassified” group (ER-/PR-/HER2-/CK5-/EGFR-). Conclusions: P-cadherin, Vimentin and CK14 can recognize BLBC already identified in triple negative/ CK5 and/or EGFR+ tumors, and due to P-cadherin sensitivity for BLBC identification this marker can reliably recruit a large number of breast carcinomas with basal phenotype among immunohistochemistry triple negative/ CK5 and/or EGFR - pool of tumors. Although they need GEP validation, our results can introduce the idea of these markers as additional options in the daily workup of breast pathology laboratories to identify BLBC.
- PublicationOpen AccessPrognostic value of p53 protein expression and clinicopathological factors in infiltrating ductal carcinoma of the breast. A study of 192 patients(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2001) Sirvent, J. J.; Fortuño Mar, A.; Olana, M.; Ortí, A.The p53 gene is located on the short arm of chromosome 17. It encodes a 53-kd nuclear protein (p53) found in scant amounts in normal tissue. Mutations of the p53 gene have been reported in different human tumours. In breast cancer, it has been noted that the overexpression of p53 protein in the nucleus is an indicator of poor prognosis, although there is a high degree of variability, which may be due to different immunohistochemical techniques, varying assessment of results a nd the type of monoclonal antibody used. This study is an immunohistochemical analysis of p53 expression in 192 cases of infiltrating ductal carcinoma of the breast, correlating it with clinicopathological factors and the clinical course of the disease. Of all the breast-cancer tissue analysed, stains for p53 antibody were found in 87 tumours (45.3%). The results of multivariate ana lysis show that the independent predictors related to recurrence are tumour size, Iymphnode metastasis and p53, while those related to death are necrosis, lymph-node metastasis and p53. In summary, our series showed prognostic significance between the expression of p53 and shorter survival time and disease-free interval for all patients in general as well as those who prese nted lymph-node metastases at the time of diagnosis.
- PublicationOpen AccessThe postulated mechanism of the protective effect of ginger on the aspirin induced gastric ulcer: Histological and immunohistochemical studies(Universidad de Murcia. Departamento de Biología Celular e Histología, 2015) Salah Khalil, MahmoudThere are many available drugs for treating gastric ulcer, but they have various side effects. Ginger is a folk, herbal medicine, which is used for treatment of various diseases including gastric ulcer. This study investigates the possible mechanism of the protective effect of ginger on aspirin induced gastric ulcer. Forty adult male albino rats were randomized into four groups (10 animals per each group) and orally received the followings once daily for 5 days: Group I: 3 ml of 1% carboxymethyl cellulose; Group II: ginger powder (200 mg/kg body weight) suspended in 3 mL of 1% carboxymethylcellulose; Group III: aspirin (400 mg/kg body weight) suspended in 3 ml of 1% carboxymethylcellulose in water. Group IV: ginger and 30 minutes later, received aspirin suspended in 1% carboxymethylcellulose, in similar doses as received in groups II and III. On day 6, rats were sacrificed. The animals were anesthetized and the stomach was removed for the macroscopic, histological (Haematoxylin and Eosin and Periodic Acid Shiff) and immunohistochemical investigations (Bax, inducible nitric oxide synthase and heat shock protein 70). Aspirin induced a significant increase of the macroscopic ulcer score, shed and disrupted epithelium, mucosal hemorrhage, submucosal edema and leukocyte infiltration, loss of the mucus of the mucosal surface significantly increased expression of apoptosis regulator Bax, inducible nitric oxide synthase (iNOS) and heatshock protein 70 (HSP70). Ginger ameliorated the histological changes by reducing Bax and iNOS and increasing HSP70 expressions.