Laboratory of Pathological anatomy

Martin, Slovakia

Laboratory of Pathological anatomy

Martin, Slovakia
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Marinowic D.R.,Pontifical Catholic University of Rio Grande do Sul | Marinowic D.R.,Institute for Biomedical Research | Majolo F.,Institute for Biomedical Research | Sebben A.D.,Pontifical Catholic University of Rio Grande do Sul | And 12 more authors.
Molecular Medicine Reports | Year: 2017

Focal cortical dysplasia (FCD) is caused by numerous alterations, which can be divided into abnormalities of the cortical architecture and cytological variations; however, the exact etiology of FCD remains unknown. The generation of induced pluripotent stem cells (iPSCs) from the cells of patients with neurological diseases, and their subsequent tissue-specific differentiation, serves as an invaluable source for testing and studying the initial development and subsequent progression of diseases associated with the central nervous system. A total of 2 patients demonstrating seizures refractory to drug treatment, characterized as FCD Type IIb, were enrolled in the present study. Fibroblasts were isolated from residual skin fragments obtained from surgical treatment and from brain samples obtained during surgical resection. iPSCs were generated following exposure of fibroblasts to viral vectors containing POU class 5 homeobox 1 (OCT4), sex determining region Y-box 2 (SOX2), Kruppel-like factor 4 and c-MYC genes, and were characterized by immunohistochemical staining for the pluripotent markers homeobox protein NANOG, SOX2, OCT4, TRA1-60 and TRA1-81. The brain samples were tested with antibodies against protein kinase B (AKT), phosphorylated-AKT, mechanistic target of rapamycin (mTOR) and phosphorylated-mTOR. Analysis of the AKT/mTOR pathway revealed a statistically significant difference between the cerebral tissues of the two patients, which were of different ages (45 and 12 years old). Clones with the morphological features of embryonic cells were detected on the 13th day and were characterized following three subcultures. The positive staining characteristics of the embryonic cells confirmed the successful generation of iPSCs derived from the patients' fibroblasts. Therefore, the present study presents a method to obtain a useful cellular source that may help to understand embryonic brain development associated with FCD.

Bono Battistoni M.F.,National University of Santa | Orcellet V.,National University of Santa | Peralta J.L.,National University of Santa | Marengo R.,National University of Santa | And 7 more authors.
Veterinary Parasitology: Regional Studies and Reports | Year: 2016

An Argentinian Dogo which suffered from anorexia, lymphadenopathy, cachexia and paresis of the hind limbs was diagnosed with trypanosomiasis in Argentina in 2013. In this study, we describe the clinical profile and its evolution as well as the molecular method employed to identify and quantify Trypanosoma evansi. © 2016 Elsevier B.V.

Adamkov M.,Comenius University | Furjelova M.,Comenius University | Bencat M.,Laboratory of Pathological Anatomy | Kruzliak P.,Masaryk University
Acta Histochemica | Year: 2014

Mismatch repair genes (MMR) play an essential role in DNA repair. MMR mutations predominantly in MLH1, MSH2, MSH6, PMS2, and rarely in PMS1, may cause the production of abnormally short or inactivated proteins. The antiapoptotic protein survivin functions in the inhibition of apoptosis, regulation of cell division and also enhances angiogenesis. Both MMRP and survivin are considered to be powerful prognostic parameters. This study was designed to determine the relationship between MMRP and survivin in colon lesions. The study included 113 cases of colon carcinoma and 51 cases of colon polyps. Survivin expression and MMRP status were assessed by immunohistochemistry. In each section, expression, intensity of immunostaining and percentage of labeled cells were analyzed. In carcinomas, immunoreaction was detected in 100/113 cases for MLH1 (88.5%), 112/113 cases for MSH2 (99.1%), 110/113 cases for MSH6 (97.3%), and 103/113 cases for PMS2 (91.2%). Survivin was shown in 47/113 cases (41.6%). The statistical analysis confirmed a significant correlation between the expression of MMRP and survivin in the assessed parameters. All 51 polyp samples were positive for MLH1, MSH2, MSH6 and PMS2. Only 8 of those (15.7%) were positive for survivin. Statistically significant differences were observed between the expression of MMRP and survivin. In conclusion, this study revealed that MMRP may suppress the antiapoptotic function of survivin through p53 inactivation of its promoter in grade 1 and grade 2 colon carcinomas. © 2014 Elsevier GmbH.

Chevalier N.,Center Hospitalier University rchet | Chevalier N.,University of Nice Sophia Antipolis | Barlier A.,Center Hospitalier University La Conception | Roche C.,Center Hospitalier University La Conception | And 10 more authors.
International Journal of Andrology | Year: 2010

Testicular germ cell cancers are the most common solid malignancies in young men, but their pathogenesis remains undetermined although some epidemiological data have implicated both environmental and genetic factors. Glial cell-derived neurotrophic factor (GDNF) is secreted by Sertoli cells, and promotes germ stem cell proliferation by activating RET, a tyrosine kinase receptor. Over-expression of GDNF in adult transgenic mice induces the development of testicular tumours that mimic human seminoma, the most frequent testicular germ cell tumour. Activating mutations of RET were previously reported in several types of cancer, including thyroid, pituitary, adrenal and melanoma cancer. Both mouse experimental model and clinical studies suggested that mutations or selective polymorphisms of RET might be associated with human seminoma. To verify this hypothesis, we conducted this study in a French University Hospital and carried out an association study using tissue samples from 66 paraffin-embedded seminoma tumours. The most frequently mutated exons of the RET proto-oncogene were sequenced to identify mutations or selective polymorphisms. No somatic mutations were identified. The polymorphic variants frequencies did not differ from those in a control Caucasian population. Human classical seminoma that occurs in young men does not appear to be linked with mutations or relevant polymorphisms of RET. © 2010 The Authors. International Journal of Andrology © 2010 European Academy of Andrology.

Adamkov M.,Comenius University | Halasova E.,Comenius University | Rajcani J.,Laboratory of Pathological anatomy | Bencat M.,Laboratory of Pathological anatomy | And 4 more authors.
Medical Science Monitor | Year: 2011

Background: The tumor suppressor gene p53 is a key regulator of cell division and/or apoptosis. Survivin is a multifunctional member of the inhibitor of apoptosis family. Survivin and p53 represent diametrically opposed signals that influence the apoptotic pathway. Material/Methods: To determine the role of p53 and survivin in basal cell carcinoma (BCC), we evaluated the expression pattern of both proteins with regard to the percentage of positively immunostained tumor cells, the intensity of staining, and subcellular localization among 31 subjects with BCC. Results: Overexpression of p53 protein was found in 28 of 31 cases (90.3%), whereas survivin accumulation was seen in 27 (87.1%). For p53, moderate and/or strong immunoreactivity was seen in 20 of 28 cases (71.4%), and 26 of 28 cases (92.9%) showed more than 25% reactive tumor cells. Nuclear p53 staining was detected in 23 of 28 cases (82.1%), whereas combined nuclear and cytoplasmic localization was found in only 5 of 28 cases (17.9%). Survivin revealed mild intensity of immunoreaction in 22 of 27 cases (71%), and 25 of 27 cases (92.6%) showed less than 25% labeled tumor cells. Combined nuclear and cytoplasmic survivin localization was present in 26 of 27 cases (96.3%). Statistically significant differences were detected in the assessed expression parameters between those proteins. Conclusions: Our results suggest that overexpression of wild type p53 protein may suppress the expression of survivin and its antiapoptotic activity in BCC cells. © Med Sci Monit.

Adamkov M.,Comenius University | Halasova E.,Comenius University | Kajo K.,Biocyt Diagnostic Center | Machalekova K.,Biocyt Diagnostic Center | And 3 more authors.
Neoplasma | Year: 2010

The antiapoptotic protein survivin can be detected in most types of malignant tumors, but it is rarely expressed in corresponding normal adult tissues. Therefore, survivin appears to represent a promising diagnostic biomarker. We examined survivin expression in 13 cases of normal breast tissue, 38 cases of fibroadenomas and 80 cases of breast carcinomas by immunohistochemical staining using anti-survivin antibody (DAKO, Clone 12C4). In each section, the intensity of staining, percentage of labeled cells, and the subcellular location of survivin antigen were assessed. Survivin was detected in 4/13 cases of normal breast tissue (30.7%), in 28/38 cases of fibroadenomas (73.7%), and in 67/80 cases of carcinomas (83.8%). Normal breast tissue showed cytoplasmic positivity only. In fibroadenomas, 19 cases (50.0%) revealed cytoplasmic reaction, and in 9 cases (23.7%), small foci of cells with combined nuclear and cytoplasmic location were identified. In carcinomas, cytoplasmic staining was found in 12/80 cases (15.0%), nuclear staining in 10/80 cases (12.5%), and combined cytoplasmic and nuclear staining in 45/80 cases (56.3%). Subcellular location of survivin between benign and malignant lesions revealed significant differences (p<0,001). Our findings point at practical use of survivin detection. We confirm the importance of nuclear staining of survivin antigen in breast carcinoma, which seems to be a notable diagnostic marker for estimation of the degree of neoplasia.

Adamkov M.,Comenius University | Kajo K.,Biocyt Diagnostic Center | Vybohova D.,Comenius University | Krajcovic J.,Comenius University | And 2 more authors.
Neoplasma | Year: 2012

The antiapoptotic protein survivin is widely expressed in most human cancers, including carcinomas of the breast. It is rarely detected in corresponding normal adult tissues. Therefore, survivin comes into the limelight as a promising diagnostic biomarker and prognostic parameter. Immunohistochemically, we examined the expression of this protein in 126 cases of ductal breast carcinoma to determine the association with clinicomorphological parameters such as age of patients, grade, stage and size of the primary tumor, lymph node metastasis, vascular invasion as well as estrogen and progesterone status. In each section, the subcellular location of survivin antigen, the intensity of staining and the percentage of labeled cells were assessed. Overall, survivin was expressed in 111 cases (88.1%). The statistical analysis revealed a significant correlation between the nuclear location of survivin and tumor grade 3. Furthermore, a significant relation was also found between vascular invasion and nuclear and combined nuclear and cytoplasmic survivin expression, together with a higher intensity of immunoreaction. However, no significant correlations were shown with other clinicomorphological parameters, such as stage and size of the tumor, lymph node metastasis, estrogen and progesterone receptors and age. Our findings revealed that survivin was frequently overexpressed in carcinoma cells, where it was present in different subcellular compartments. The nuclear positivity of survivin or combined nuclear and cytoplasmic expression was shown to be a poor prognostic parameter in ductal breast carcinoma.

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