Histology and histopathology Vol.28, nº 3 (2013)

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  • Publication
    Open Access
    Levels of acyl-Coenzyme A synthetase 5 in urothelial cells and corresponding neoplasias reflect cellular differentiation
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Gaisa, Nadine T.; Reinartz, Andrea; Schneider, Ursula; Klaus, Christina; Heidenreich, Axel; Jakse, Gerhard; Kaemmerer, Elke; Klinkhammer, Barbara Mara; Knuechel, Ruth; Gassler, Nikolaus
    Metabolic components like fatty acids and acyl-Coenzyme A (acyl-CoA) thioesters have been implicated in the pathogenesis of various tumours. The activation of fatty acids to acyl-CoAs is catalysed by long chain acyl-CoA synthetases (ACSLs), and impairment of ACSL expression levels has been associated with tumourigenesis and progression. Since ACSLs have never been investigated in bladder tissues, the study aims to characterize ACSL expression and acyl-CoA synthesis in normal and neoplastic bladder tissues, as well as cell lines. ACSL isoforms 1, 3, 4 and 5 and synthesis of acyl-CoAs were analysed using qRTPCR, western blot analysis, immunohistochemistry and lipid mass spectrometry. In normal urothelium, expression of ACSL1, 3, 4 and 5, with highest levels of ACSL isoform 5 was found. However, ACSL5 expression was reduced in corresponding neoplastic tissues and urothelial cell lines depending on the grade of cellular differentiation. AntiACSL5 immunostainings showed expression in normal urothelium and a gradual loss of ACSL5 protein via preinvasive lesions to invasive carcinomas. High expression of ACSL5 correlated with increased α-galactosidase activity and positive Uroplakin III staining in tumours. In contrast, synthesis of acyl-CoAs was enhanced in neoplastic bladder tissues compared to normal urothelium, and reflected an increase with respect to cellular differentiation. These results confirm an expression of ACSLs, especially isoform 5, in human urothelium, prove enzymatic/lipidomic changes in bladder cancer tissues, and suggest an involvement of ACSL5 in cellular maturation and/or senescence with possible effects onto induction of tumour formation or progression. Further work may identify responsible pathway alterations, and attempting to re-balance the metabolic equilibrium of the urothelium may offer a further opportunity for tumour treatment and prevention.
  • Publication
    Open Access
    DKC1 gene mutations in human sporadic cancer
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Penzo, Marianna; Casoli, Lucia; Ceccarelli, Claudio; Treré, Davide; Ludovini, Vienna; Crinò, Lucio; Montanaro, Lorenzo
    Introduction: Germline mutations in the tumour suppressor gene dyskeratosis congenit 1 (DKC1) cause the cancer prone syndrome called X-linked dyskeratosis congenita. The present study aims to determine whether mutations of the DKC1 gene may also be present in frequent human sporadic cancers (breast, colon and lung cancers), thus potentially contributing to the neoplastic phenotype. Materials and methods: mutation analysis of the DKC1 gene was performed on DNA from 110 primary human lung, 54 breast, and 35 colon cancers, focusing on gene regions where pathogenic germline mutations have been described previously (promoter and exons 1, 3, 9, 10, 11, and 14). Results: Out of a total of 199 primary tumours of different origins, only 5 turned out to have sequence variations in the DKC1 gene. These variations were of two kinds, C8120T and C13554T, which are both classified as synonymous mutations and do not affect DKC1 mRNA splicing. Conclusion: direct DKC1 gene mutations are not a frequent event in tumourigenesis, at least in the tumour types investigated and for the DKC1 gene portions considered in this study.
  • Publication
    Open Access
    Immunohistochemical study of enteric nervous system in hirschsprung’s disease and intestinal neuronal dysplasia
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Geramizadeh, Bita; Akbarzadeh, Elahe; Izadi, Babak; Foroutan, Hamid-Reza; Heidari, Tahere
    Background. Hirschsprung’s disease (HD) is one of the most common motility disorders in pediatric age groups and it is very important that it be differentiated from other types of motility disorders, especially intestinal neuronal dysplasia B (IND B). Although many studies regarding the differences between the two disorders by immunohistochemical studies exist, there is as yet no consistent result. The purpose of this research was to study the immunohistochemical findings of enteric nervous system in these two motility disorders in comparison with colectomies without motility disorder. Methods. Full wall thickness specimens of three groups of patients (HD, IND B and non motility disorders) were included in the study to be evaluated by immunohistochemistry (IHC). Markers were specific for neuronal cells and pace maker cells composed of PGP 9.5, c-kit, synaptophysin, S100 and CD56. The number of cells was evaluated in the muscularis properia, and myenteric plexus. Results. The number of all the IHC markers i.e. PGP9.5, c-kit, synaptophysin, S100 and CD56 was completely different in HD from the two other groups, while IND B was quite similar to control group. Conclusion. Our finding suggests that there is a marked and significant difference between HD and IND B by IHC markers, which can be used as an additional test for the diagnosis of HD with more accuracy. Further multicenter studies with a greater number of cases would be necessary to find a cut-off point for every IHC marker to differentiate HD and IND B.
  • Publication
    Open Access
    Clear cell papillary cystadenocarcinoma of the epididymis: a case report and immunohistochemistry of markers for renal cell carcinoma
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Nozawa, Tatsuru; Konda, Ryuichiro; Ohsawa, Taisuke; Yoshida, Makoto; Komatsu, Masayo; Iwama, Takahide; Fujioka, Tomoaki
    Neoplasms of the epididymis are uncommon, and malignant tumors are extremely rare. We report a case of clear cell papillary cystadenocarcinoma of the epididymis presenting with a long history of painless scrotal mass on the left side. Immunohistochemical markers for clear cell renal cell carcinoma (RCC) were examined to distinguish between clear cell papillary cystadenocarcinoma of the epididymis and metastatic clear cell renal cell carcinoma. The present case was positive for cytokeratin-7, PAX2, vinculin, vimentin and carbonic anhydrase IX. Expression of CD10 was focally observed. In contrast, no immunoreactivities for α- methylacyl-CoA racemase, RCC marker, glutathione Stransferase α or C-KIT were detected. The immunophenotypic profile of clear cell papillary cystadenocarcinoma of the epididymis closely resembles that of clear cell papillary RCC, although the immunohistochemical markers tested in this study are useful to make a differential diagnosis between clear cell papillary cystadenocarcinoma of the epididymis and metastatic clear cell RCC.
  • Publication
    Open Access
    Different influence of ovine estrus synchronization treatments on caruncular early angiogenesis
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Ruiz-González, Irene; Sánchez, M.A.; García-Fernández, R.A.; García-Palencia, P.; Sánchez, B.; González-Bulnes, A.; Flores, J.M.
    The present study compares two protocols for ovine estrus synchronization by assessing the caruncular angiogenic response to the establishment of pregnancy. The analysis consisted of the immunohistochemical evaluation of Vascular Endothelial Growth Factor (VEGF), Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1, CD31) and Von Willebrand Factor (vWF) in ovine caruncular stroma. A flock of thirty-eight adult ewes was divided in two groups and synchronized with either progestagens (Group P) or prostaglandin analogues (Group PG). Immunohistochemistry was performed in uterine samples obtained from pregnant ewes (P, n=15; PG, n=13) on days 15 post coitus (pc), 17pc and 21pc (day 0 =day of estrus). Each factor was assessed by total vascular density (TVD, total positive blood vessels/mm2), capillary vascular density (CVD, positive blood capillaries/mm2) and arteriolar vascular density (AVD, positive arterioles/mm2). Group P demonstrated higher VEGF-CVD (P=0.045) when compared to prostaglandin treated animals. Vascular CD31- expression decreased on days 15pc and 21pc (TVD, P=0.007 and CVD, P=0.014) in both groups. vWF analysis did not show significant differences between groups or days of study. These results demonstrate a different influence of progestagen-based and prostaglandin analogues-based synchronization treatments over VEGF vascular expression during caruncular development taking place in response to pregnancy establishment. In addition, observations pointed out in this study support the involvement of CD31 in the angiogenic stimulus that occurs during early maternal placentation in the ewe.