Publications by authors named "Ivana Ticha"

27 Publications

  • Page 1 of 1

Healthy Food on Instagram Social Network: Vegan, Homemade and Clean Eating.

Nutrients 2021 Jun 9;13(6). Epub 2021 Jun 9.

Department of Management, Faculty of Economics and Management, Czech University of Life Sciences Prague, 165 21 Prague, Czech Republic.

Social media platforms have become part of many people's lives. Users are spending more and more time on these platforms, creating an active and passive digital footprint through their interaction. This footprint has high research potential in many research areas because understanding people's communication on social media is essential in understanding their values, attitudes, experiences and behaviors. Researchers found that the use of social networking sites impacts adolescents' eating behavior. If we define adolescents as individuals between ages 10 and 24 (WHO's definition), 76% of USA young people at age 18-⁠24 use Instagram, so the Instagram social network analysis is important for understanding young people's expressions in the context of healthy food. This study aims to identify the main topic associated with healthy food on the Instagram social network via hashtag and community analysis based on 2,045,653 messages created by 427,936 individual users. The results show that users most associate Healthy food with healthy lifestyle, fitness, weight loss and diet. In terms of food, these are foods that are Vegan, Homemade, Clean and Plant-based. Given that young people change their behavior in relation to people's behavior on social networks, it is possible to use this data to predict their future association with healthy food characteristics.
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http://dx.doi.org/10.3390/nu13061991DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8226706PMC
June 2021

Microscopic extraovarian sex cord proliferation: report of a case with bilateral Fallopian tube involvement and a comprehensive molecular analysis.

Pol J Pathol 2020 ;71(2):175-180

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic.

We report a case of a 58-year-old female with microscopic extraovarian sex cord proliferations affecting both Fallopian tubes. Molecular analysis showed likely pathogenic germline missense mutations of the KDM5A and KMT2D genes. However, mutations of other genes, including FOXL2 and STK11, were not detected. Our case represents the 12th case of extraovarian sex cord proliferation reported in the literature to date. This is the first time that a molecular genetic analysis of the lesion has been performed, and it showed a wild-type FOXL2 gene, which represents another argument supporting the estimated benign nature of these rare lesions.
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http://dx.doi.org/10.5114/pjp.2020.97023DOI Listing
October 2020

Impact of Think-Aloud on Eye-Tracking: A Comparison of Concurrent and Retrospective Think-Aloud for Research on Decision-Making in the Game Environment.

Sensors (Basel) 2020 May 12;20(10). Epub 2020 May 12.

Department of Management, Faculty of Economics and Management, Czech University of Life Sciences Prague, 165 21 Prague, Czech Republic.

Simulations and games bring the possibility to research complex processes of managerial decision-making. However, this modern field requires adequate methodological procedures. Many authors recommend the use of a combination of concurrent think-aloud (CTA) or retrospective think-aloud (RTA) with eye-tracking to investigate cognitive processes such as decision-making. Nevertheless, previous studies have little or no consideration of the possible differential impact of both think-aloud methods on data provided by eye-tracking. Therefore, the main aim of this study is to compare and assess if and how these methods differ in terms of their impact on eye-tracking. The experiment was conducted for this purpose. Participants were 14 managers who played a specific simulation game with CTA use and 17 managers who played the same game with RTA use. The results empirically prove that CTA significantly distorts data provided by eye-tracking, whereas data gathered when RTA is used, provide independent pieces of evidence about the participants' behavior. These findings suggest that RTA is more suitable for combined use with eye-tracking for the purpose of the research of decision-making in the game environment.
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http://dx.doi.org/10.3390/s20102750DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7294419PMC
May 2020

Pathologic Protocols for Sentinel Lymph Nodes Ultrastaging in Cervical Cancer.

Arch Pathol Lab Med 2019 Dec 23. Epub 2019 Dec 23.

From Institute of Pathology (Drs Dundr, Němejcová, Tichá, Bártů, and Jakša) and Gynecologic Oncology Center, Department of Obstetrics and Gynecology (Dr Cibula), First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic.

Context.—: Ultrastaging of sentinel lymph nodes (SLNs) is a crucial aspect in the approach to SLN processing. No consensual protocol for pathologic ultrastaging has been approved by international societies to date.

Objective.—: To provide a review of the ultrastaging protocol and all its aspects related to the processing of SLNs in patients with cervical cancer.

Data Sources.—: In total, 127 publications reporting data from 9085 cases were identified in the literature. In 24% of studies, the information about SLN processing is entirely missing. No ultrastaging protocol was used in 7% of publications. When described, the differences in all aspects of SLN processing among the studies and institutions are substantial. This includes grossing of the SLN, which is not completely sliced and processed in almost 20% of studies. The reported protocols varied in all aspects of SLN processing, including the thickness of slices (range, 1-5 mm), the number of levels (range, 0-cut out until no tissue left), distance between the levels (range, 40-1000 μm), and number of sections per level (range, 1-5).

Conclusions.—: We found substantial differences in protocols used for SLN pathologic ultrastaging, which can impact sensitivity for detection of micrometastases and even small macrometastases. Since the involvement of pelvic lymph nodes is the most important negative prognostic factor, such profound discrepancies influence the referral of patients to adjuvant radiotherapy and could potentially cause treatment failure. It is urgent that international societies agree on a consensual protocol before SLN biopsy without pelvic lymphadenectomy is introduced into routine clinical practice.
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http://dx.doi.org/10.5858/arpa.2019-0249-RADOI Listing
December 2019

A comprehensive evaluation of pathogenic mutations in primary cutaneous melanomas, including the identification of novel loss-of-function variants.

Sci Rep 2019 11 19;9(1):17050. Epub 2019 Nov 19.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

The most common histological subtypes of cutaneous melanoma include superficial spreading and nodular melanoma. However, the spectrum of somatic mutations developed in those lesions and all potential druggable targets have not yet been fully elucidated. We present the results of a sequence capture NGS analysis of 114 primary nodular and superficial spreading melanomas identifying driver mutations using biostatistical, immunohistochemical and/or functional approach. The spectrum and frequency of pathogenic or likely pathogenic variants were identified across 54 evaluated genes, including 59 novel mutations, and the newly identified TP53 loss-of-function mutations p.(L194P) and p.(R280K). Frequently mutated genes most commonly affected the MAPK pathway, followed by chromatin remodeling, and cell cycle regulation. Frequent aberrations were also detected in the genes coding for proteins involved in DNA repair and the regulation and modification of cellular tight junctions. Furthermore, relatively frequent mutations were described in KDR and MET, which represent potential clinically important targets. Those results suggest that with the development of new therapeutic possibilities, not only BRAF testing, but complex molecular testing of cutaneous melanoma may become an integral part of the decision process concerning the treatment of patients with melanoma.
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http://dx.doi.org/10.1038/s41598-019-53636-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6863855PMC
November 2019

Truncated PPM1D impairs stem cell response to genotoxic stress and promotes growth of APC-deficient tumors in the mouse colon.

Cell Death Dis 2019 10 28;10(11):818. Epub 2019 Oct 28.

Cancer Cell Biology, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czech Republic.

Protein phosphatase magnesium-dependent 1 delta (PPM1D) terminates cell response to genotoxic stress by negatively regulating the tumor suppressor p53 and other targets at chromatin. Mutations in the exon 6 of the PPM1D result in production of a highly stable, C-terminally truncated PPM1D. These gain-of-function PPM1D mutations are present in various human cancers but their role in tumorigenesis remains unresolved. Here we show that truncated PPM1D impairs activation of the cell cycle checkpoints in human non-transformed RPE cells and allows proliferation in the presence of DNA damage. Next, we developed a mouse model by introducing a truncating mutation in the PPM1D locus and tested contribution of the oncogenic PPM1D allele to colon tumorigenesis. We found that p53 pathway was suppressed in colon stem cells harboring PPM1D resulting in proliferation advantage under genotoxic stress condition. In addition, truncated PPM1D promoted tumor growth in the colon in Apc mice and diminished survival. Moreover, tumor organoids derived from colon of the ApcPpm1d mice were less sensitive to 5-fluorouracil when compared to ApcPpm1dand the sensitivity to 5-fluorouracil was restored by inhibition of PPM1D. Finally, we screened colorectal cancer patients and identified recurrent somatic PPM1D mutations in a fraction of colon adenocarcinomas that are p53 proficient and show defects in mismatch DNA repair. In summary, we provide the first in vivo evidence that truncated PPM1D can promote tumor growth and modulate sensitivity to chemotherapy.
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http://dx.doi.org/10.1038/s41419-019-2057-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817818PMC
October 2019

Impact of chemotherapy on the expression of claudins and cadherins in invasive breast cancer.

Exp Ther Med 2019 Oct 20;18(4):3014-3024. Epub 2019 Aug 20.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, 12800 Prague, Czech Republic.

The importance of the expression profile of claudins in the molecular classification of breast cancer (BC) is currently under investigation. Claudins, together with cadherins, serve an important role in the epithelial-mesenchymal transition and influence the chemosensitivity of cancer cells. Adjuvant chemotherapy is administered following surgical resection in selected cases of BC. Previous neoadjuvant chemotherapy may change the molecular profile of a tumour and subsequently also its chemosensitivity. In the current study, the expression of claudin-1, -3 and -4, E- and N-cadherin and the standard BC biomarkers [oestrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and marker of proliferation Ki-67 (Ki-67)] in formalin-fixed, paraffin-embedded sections from 62 patients with invasive BC was analysed using immunohistochemistry prior to and following neoadjuvant chemotherapy. The results revealed increased expression of claudin-1 (P=0.03) and decreased expression of claudin-3 (P=0.005), PR (P<0.001) and Ki-67 (P=0.01) following the neoadjuvant therapy. No significant changes in the expression of ER, claudin-4 or E- and N-cadherin were observed following therapy. Furthermore, an association between the expression of claudin-1 and the standard BC markers (P<0.05) was identified. A high expression of claudin-1 was more frequently observed in the triple-negative BC cohort than in the cohort with positive ER, PR and/or HER2 before (P=0.04) and after chemotherapy (P=0.02). The expression of N-cadherin was associated with the expression of ER, PR, HER2 and tumour grade (P<0.05). A positive association between the expression of claudin-3 and E-cadherin (P=0.005) was observed. No association was found between the expression of E- and N-cadherin. In conclusion, significant changes in the expression of claudin-1 and -3 but not in the expression of claudin-4, E- and N-cadherin were observed in samples taken from patients with BC following chemotherapy. These findings indicate that claudins-1 and -3 serve a role in the response of BC to chemotherapy.
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http://dx.doi.org/10.3892/etm.2019.7930DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6755479PMC
October 2019

Leiomyoma with Bizarre Nuclei: a Study of 108 Cases Focusing on Clinicopathological Features, Morphology, and Fumarate Hydratase Alterations.

Pathol Oncol Res 2020 Jul 31;26(3):1527-1537. Epub 2019 Aug 31.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Studnickova 2, 12800, Prague 2, Czech Republic.

Leiomyoma with bizarre nuclei (LBN) is an uncommon variant of uterine smooth muscle neoplasm. Involvement of fumarate hydratase (FH) has been suggested in the pathogenesis of a subset of LBN. The goal of our study is to assess the clinicopathological, morphological, immunohistochemical and molecular findings focusing on FH in LBNs (n = 108) and compare it with the findings in usual leiomyomas (UL; n = 50) and leiomyosarcomas (LMS; n = 42). Immunohistochemically, loss of FH expression was found in 67/108 of LBN, 1/50 of UL and in no LMS. Class 4/5 FH mutations were detected in 15/53 LBN with sufficient DNA quality for molecular analysis. Pathogenic variants of the FH gene were detected in neither UL nor LMS. Local recurrence after surgery was present in 18/92 of LBN patients, 7 of which were histologically verified and 2 of which were found to be LBN. Our results confirmed that LBN behave in a benign fashion, although they may relapse. FH gene mutations were a common finding only in LBN, but not in UL and LMS. Immunohistochemistry with an antibody against FH seems to have a good sensitivity (87%) and moderate specificity (58%) with regard to predicting FH gene mutations and could be used as a screening method in tumors with features suggestive of FH alterations to identify patients who are at risk for the FH aberrations.
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http://dx.doi.org/10.1007/s12253-019-00739-5DOI Listing
July 2020

Synchronous endometrioid endometrial and ovarian carcinomas are biologically related: A clinico-pathological and molecular (next generation sequencing) study of 22 cases.

Oncol Lett 2019 Feb 20;17(2):2207-2214. Epub 2018 Dec 20.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, 12800 Prague, Czech Republic.

The criteria for distinction between independent primary tumors and metastasis from one site to the other in synchronous endometrioid endometrial and ovarian carcinoma (SEO) has been a matter of dispute for a long time. In our study we performed a comprehensive clinico-pathological and molecular analysis of 22 cases of SEO. Based on conventional clinico-pathological criteria the cases were classified as independent primary tumors (10 cases) and metastasis from one location to the other (12 cases). All tumors were analyzed by NGS with a panel of 73 genes (219 kbp). Clonal origin was confirmed in all cases by at least one shared mutation in and . Two patients carried germline pathogenic mutation in cancer-predisposing genes or . Microsatellite instable phenotype was detected in 5/22 (22.7%) SEO, but in one case only in the endometrial tumor. In conclusion, our results showed that all 22 SEOs were clonally related, irrespectively of their clinico-pathological features. Even low grade and low stage tumors classified as independent primaries, according to the conventional morphological criteria, have a clonal origin. From the practical point of view, only the conventional morphological criteria should be used for the classification (staging) of these tumors. However, molecular profiling of these tumors may have prognostic and predictive meaning.
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http://dx.doi.org/10.3892/ol.2018.9855DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6341770PMC
February 2019

Detection of EGFR Mutations in Circulating Tumor DNA (ctDNA) Retrieved from Plasma - Interlaboratory Quality Assessment in the Czech Republic.

Klin Onkol Fall 2018;31(5):353-360

Background: Detection of EGFR mutations in tumor tissue represents a standard testing procedure in patients with non-small cell lung cancer. Molecular testing of circulating tumor DNA (ctDNA) in plasma enables detection of mutations in cases where tumor specimens are unavailable or when monitoring of therapeutic responses is necessary. In addition, according to the recent literature, ctDNA better reflects the heterogeneity of the neoplastic cell population than isolated tumor lesion or metastasis. We report a national interlaboratory evaluation aimed at assessing the analytical quality of ctDNA EGFR testing in plasma across seven reference laboratories in the Czech Republic.

Material And Methods: Aliquots of 13 plasma samples were sent to 7 laboratories and consisted of commercially available 2ml plasma specimens of genomic DNA with mutant allelic frequencies of 5, 0.5, 0.05, and 0% of the most common sensitizing mutations (deletion in exon 19, L858R) and the resistance mutation T790M. DNA extraction and EGFR testing were performed according to standard procedures. In 6/7 laboratories the cobas® EGFR Mutation Test v2 was used. One laboratory employed the Super-ARMS® EGFR Mutation Detection Kit.

Results: In total, 91 genotypes were determined with an overall error rate of 24.2% (22/91). The overall error rates were 3.2% (2/63) for the 0.5% mutation frequency and 0% for the 5% mutation frequency (0/35), respectively. No false positive results were reported. The cobas® method achieved consistent results with the 0.05% mutation frequency for the exon 19 deletion. For L858R and T790M mutations, the threshold was above the 0.5% frequency.

Conclusions: The results show that EGFR testing for ctDNA in plasma has limited sensitivity, especially for detection of the T790M mutation. Particularly, in ctDNA testing of very low mutated DNA plasma fractions (below 0.01%), emphasis should be placed on the use of highly sensitive molecular methods. The outcomes of this quality assessment confirm the need for rebiopsy in patients with negative plasma results because of a higher false negative rate in comparison to tissue testing. Key words: circulating DNA - liquid biopsy - epidermal growth factor receptor - non-small cell lung cancer - quality control This work was supported by grants of AstraZeneca and the project of the Ministry of Health number 00064203 (Motol University Hospital). The authors declare they have no potential confl icts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 4. 6. 2018 Accepted: 1. 8. 2018.
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http://dx.doi.org/10.14735/amko2018353DOI Listing
August 2019

Comparison of five different scoring methods in the evaluation of inflammatory infiltration (tumor-infiltrating lymphocytes) in superficial spreading and nodular melanoma.

Pigment Cell Melanoma Res 2019 05 21;32(3):412-423. Epub 2018 Dec 21.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

The objective of our study was to compare the five different scoring methods of tumor-infiltrating lymphocytes (TILs) assessment in a group of 213 cases of superficial spreading and nodular melanoma. The scoring methods include (a) Clark scoring; (b) Melanoma Institute Australia system; (c) scoring system used in the study of Saldanha et al.; (d) scoring system used in the TCGA study and modified by Park et al.; and (e) the system recently proposed by the "International Immuno-Oncology Biomarker Working Group" for TILs scoring in all solid tumors. Prediction of survival with three main outcomes-disease-specific-free survival, local recurrence-free survival, and distant metastasis-free survival-was evaluated. The prognostic value of TILs showed statistical significance in univariate analysis regarding all three of the outcomes only for three of the five evaluated methods; the Clark scoring, the Melanoma Institute Australia system, and the system proposed by the "International Immuno-Oncology Biomarker Working Group". However, in multivariate analysis with covariants including Breslow thickness, type of melanoma, location, sex, and age, we did not find TILs to be an independent prognostic factor.
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http://dx.doi.org/10.1111/pcmr.12757DOI Listing
May 2019

Stathmin is a potential therapeutic target but not a prognostic marker in melanoma: an immunohistochemical study of 323 melanocytic lesions.

Melanoma Res 2019 04;29(2):157-162

Department of Dermatology and Venereology, First Faculty of Medicine, Charles University and General University Hospital in Prague.

In several solid tumors, an increased stathmin expression is associated with both poor prognosis and resistance to certain chemotherapy types. However, the data regarding melanocytic lesions are very limited. The goals of our study are as follows: the assessment of stathmin expression in benign and malignant melanocytic lesions, and the significance of its expression for the differential diagnostics between benign and malignant lesions; the analysis of the prognostic significance of stathmin expression in melanoma; and the evaluation of stathmin expression in melanoma and melanoma metastases with respect to possible therapeutic targeting. Immunohistochemical analysis of stathmin expression was done in 323 melanocytic lesions, including 205 primary cutaneous melanomas, 60 melanoma metastases, and 58 melanocytic nevi. Stathmin expression was found in all analyzed groups of melanocytic lesions. Using the H-scoring system, the observed intensity of expression was as follows: melanocytic nevi: 146.1 (mean) and 150 (median); melanomas: 116.7 (mean) and 110 (median); and melanoma metastases: 136.8 (mean) and 140 (median). The stathmin expression was significantly lower in the cohort of primary melanomas when compared with metastases and nevi (P=0.001). The stathmin expression showed no prognostic significance. The high stathmin expression in melanoma suggests that stathmin might be a promising marker for therapeutic targeting in ongoing clinical trials. Compared with several other solid tumors, stathmin expression in melanoma showed no prognostic significance. The potential use of stathmin expression in differential diagnostics is limited by its common expression, and despite the statistically significant differences between nevi and melanoma, it may not be used in this setting.
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http://dx.doi.org/10.1097/CMR.0000000000000550DOI Listing
April 2019

Lymphoepithelioma-like carcinoma of the endometrium: Case report of a rare tumour with comprehensive immunohistochemical and molecular analysis.

Pol J Pathol 2018;69(1):87-92

We are reporting a case of endometrial lymphoepithelioma-like carcinoma (LELC) in a 63-year-old female. Microscopically, the tumor consisted of groups of tumor cells surrounded by dense lymphoplasmacytic infiltrate. Immunohistochemically, the tumour cells were positive for cytokeratins AE1/AE3, EMA, PAX8, p16, and estrogen receptors. Protein p53 showed an aberrant type of expression. Molecular genetic analysis revealed mutations in the TP53 and PIKP53CA genes. Based on our results, we believe that the tumor represents an unusual morphological variant of endometrial serous carcinoma.To the best of our knowledge, only six cases of LELC arising in endometrium have been reported in literature to date.
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http://dx.doi.org/10.5114/pjp.2018.75342DOI Listing
July 2018

Germline mutation in the TP53 gene in uveal melanoma.

Sci Rep 2018 05 16;8(1):7618. Epub 2018 May 16.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

We performed comprehensive molecular analysis of five cases of metastasizing uveal malignant melanoma (UM) (fresh-frozen samples) with an NGS panel of 73 genes. A likely pathogenic germline TP53 mutation c.760A > G (p.I254V) was found in two tumor samples and matched nontumor tissue. In three cases, pathogenic BAP1 mutation was detected together with germline missense variants of uncertain significance in ATM. All cases carried recurrent activating GNAQ or GNA11 mutation. Moreover, we analyzed samples from another 16 patients with primary UM by direct Sanger sequencing focusing only on TP53 coding region. No other germline TP53 mutation was detected in these samples. Germline TP53 mutation, usually associated with Li-Fraumeni syndrome, is a rare event in UM. To the best of our knowledge, only one family with germline TP53 mutation has previously been described. In our study, we detected TP53 mutation in two patients without known family relationship. The identification of germline aberrations in TP53 or BAP1 is important to identify patients with Li-Fraumeni syndrome or BAP1 cancer syndrome, which is also crucial for proper genetic counseling.
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http://dx.doi.org/10.1038/s41598-018-26040-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5955881PMC
May 2018

[Evaluation of inflammatory cells (tumor infiltrating lymphocytes - TIL) in malignant melanoma].

Cesk Patol Spring 2018;54(1):27-31

The evaluation of inflammatory infiltrate (tumor infiltrating lymphocytes - TIL) should be a standard part of biopsy examination for malignant melanoma. Currently, the most commonly used assessment method according to Clark is not optimal and there have been attempts to find an alternative system. Here we present an overview of possible approaches involving five different evaluation methods based on hematoxylin-eosin staining, including the recent suggestion of unified TIL evaluation method for all solid tumors. The issue of methodology, prognostic and predictive significance of TIL determination as well as the importance of immunohistochemical subtyping of inflammatory infiltrate is discussed.
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April 2019

Expression of Glut-1 in Malignant Melanoma and Melanocytic Nevi: an Immunohistochemical Study of 400 Cases.

Pathol Oncol Res 2019 Jan 11;25(1):361-368. Epub 2017 Nov 11.

Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Studnickova 2, 12800, Prague 2, Czech Republic.

The glucose transporter-1 (Glut-1) is a cell membrane glycoprotein involved in glucose uptake. An increased expression of Glut-1 is an important cell adaptation mechanism against hypoxia. An upregulation of Glut-1 can be found in several types of malignant tumors, which are able to reprogram their metabolism from oxidative phosphorylation to aerobic glycolysis (Warburg effect). However, the data regarding melanocytic lesions is equivocal. We performed comprehensive immunohistochemical analysis of the Glut-1 expression in 225 malignant melanomas (MM) and 175 benign nevi. Only the membranous expression of Glut-1 was regarded as positive. The expression of Glut-1 (the cut-off for positivity was determined as H-score 15) was found in 69/225 malignant melanomas. The number of positive cases and the H-score of Glut-1 increased where there was a higher Breslow thickness (p < 0.00001) when comparing pT1- pT4 MM groups. All benign nevi were classified as negative. In conclusion, the membranous expression of Glut-1 is a common feature of a malignant melanoma but this type of expression is very rare in benign melanocytic nevi. Our results suggest that the membranous expression of Glut-1 can be used as a surrogate marker in the assessing of the biological nature of benign and malignant melanocytic lesions. However, despite its high specificity, the sensitivity of this marker is relatively low. Moreover, due to the fact that the increased expression of Glut-1 correlates with a shorter survival period (10-year disease free survival, recurrence free survival and metastasis free survival and MFS), it can be used as a prognostically adverse factor.
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http://dx.doi.org/10.1007/s12253-017-0363-7DOI Listing
January 2019

Expression of Glut-1 in Normal Endometrium and Endometrial Lesions: Analysis of 336 Cases.

Int J Surg Pathol 2017 Aug 18;25(5):389-396. Epub 2016 Dec 18.

1 Department of Pathology, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.

Background: Glucose transporter-1 (Glut-1) is a membrane glycoprotein that is, together with other glucose transporters, responsible for the regulation of glucose uptake. An increased expression of this protein seems to be a general feature of several malignant tumors that are able to reprogram their metabolism and switch from oxidative phosphorylation to aerobic glycolysis.

Methods: We performed comprehensive immunohistochemical analysis of Glut-1 expression in 336 endometrial samples, including tumors, nontumor lesions, and normal tissues.

Results: Expression of Glut-1 was found in 87% of endometrioid carcinomas (160/184 cases), 100% of serous carcinomas (29/29 cases), 100% of clear cell carcinomas (17/17 cases), 50% of polyps with atypical hyperplasia (8/16 cases), 12.5% of polyps with non-atypical hyperplasia (3/24 cases), 77% of hyperplasias with atypias (10/13 cases), 9% of hyperplasias without atypias (1/11 cases), 87% of secretory endometrium samples (13/15 cases), and in none of the nonsecretory endometrium samples (0/27 cases). In endometrioid carcinomas, Glut-1 was expressed in a marked geographical pattern. In nontumor lesions, its expression was more common in atypical hyperplasia and polyps with atypical hyperplasia compared with polyps with non-atypical hyperplasia and hyperplasias without atypia ( P = .00032).

Conclusion: Our study confirms the high expression of Glut-1 not only in endometrioid carcinomas but also in other carcinomas of endometrium including clear cell and serous types. Glut-1 expression can be used as a surrogate marker in differential diagnosis between hyperplasia with and without atypia. Because of common Glut-1 expression in malignant tumors, therapeutic strategies influencing this protein or its signaling pathways can be beneficial.
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http://dx.doi.org/10.1177/1066896916683510DOI Listing
August 2017

The c.657del5 variant in the NBN gene predisposes to pancreatic cancer.

Gene 2016 Aug 2;587(2):169-72. Epub 2016 May 2.

Institute of Biochemistry and Experimental Oncology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic. Electronic address:

Pancreatic ductal adenocarcinoma (PDAC) is the sixth most frequent cancer type in the Czech Republic with a poor prognosis that could be improved by an early detection and subsequent surgical treatment combined with chemotherapy. Genetic factors play an important role in PDAC risk. We previously identified one PDAC patient harboring the Slavic founder deleterious mutation c.657del5 in the NBN gene, using a panel next-generation sequencing (NGS). A subsequent analysis of 241 unselected PDAC patients revealed other mutation carriers. The overall frequency of c.657del5 in unselected PDAC patients (5/241; 2.07%) significantly differed from that in non-cancer controls (2/915; 0.2%; P=0.006). The result indicates that the NBN c.657del5 variant represents a novel PDAC-susceptibility allele increasing PDAC risk (OR=9.7; 95% CI: 1.9 to 50.2). The increased risk of PDAC in follow-up recommendations for NBN mutation carriers should be considered if other studies also confirm an increased frequency of c.657del5 carriers in PDAC patients from other populations.
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http://dx.doi.org/10.1016/j.gene.2016.04.056DOI Listing
August 2016

MTDH genetic variants in colorectal cancer patients.

Sci Rep 2016 Mar 17;6:23163. Epub 2016 Mar 17.

Department of Oncology and Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.

The colorectal carcinogenesis is a complex process encompassing genetic alterations. The oncoprotein AEG-1, encoded by the MTDH gene, was shown previously to be involved in colorectal cancer (CRC). The aim of this study was to determine the frequency and the spectrum of MTDH variants in tumor tissue, and their relationship to clinicopathological variables in CRC patients. The study included tumors from 356 unselected CRC patients. Mutation analysis of the MTDH gene, including coding region and adjacent intronic sequences, was performed by direct DNA sequencing. The corresponding normal colorectal tissue was analyzed in the carriers of exonic variant to confirm germline or somatic origin. We detected 42 intronic variants, where 25 were novel. Furthermore, we found 8 exonic variants of which four, one missense (c.977C > G-germline) and three frameshift mutations (c.533delA-somatic, c.1340dupA-unknown origin, c.1731delA-unknown origin), were novel. In silico prediction analyses suggested four deleterious variants (c.232G > T, c.533delA, c.1340dupA, and c.1731delA). There were no correlations between the MTDH variants and tumor stage, differentiation or patient survival. We described several novel exonic and intronic variants of the MTDH gene. The detection of likely pathogenic truncating mutations and alterations in functional protein domains indicate their clinical significance, although none of the variants had prognostic potential.
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http://dx.doi.org/10.1038/srep23163DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4794727PMC
March 2016

Expression, Epigenetic and Genetic Changes of HNF1B in Endometrial Lesions.

Pathol Oncol Res 2016 Jul 19;22(3):523-30. Epub 2015 Dec 19.

Department of Pathology, First Faculty of Medicine and General University Hospital, Charles University in Prague, Studnickova 2, 2 12800, Prague, Czech Republic.

Hepatocyte nuclear factor 1-beta (HNF-1-beta) is a transcription factor involved in cancerogenesis of various tumors, including endometrioid carcinoma. We performed comprehensive analysis of HNF-1-beta in lesions of the endometrium, including protein expression and genetic and epigenetic changes. Expression of HNF-1-beta was analyzed immunohistochemically in 320 cases including both tumor and non-tumor endometrial lesions. Promoter methylation and genetic variants were evaluated, using bisulphite and direct sequencing, in 30 (18 fresh frozen, 12 FFPE tumors) endometrioid carcinomas (ECs) and 15 ovarian clear cell carcinomas (OCCCs) as a control group. We detected expression of HNF-1-beta in 28 % of ECs (51/180 cases), 26 % of serous carcinoma (7/27 cases), 83 % of endometrial clear cell carcinoma (15/18 cases), 93 % of hyperplastic polyps with atypias (13/14 cases), 100 % of hyperplastic polyps without atypias (16/16 cases), 88 % of hyperplasias with atypias (14/16 cases), 91 % of hyperplasias without atypias (10/11 cases), and in ≥80 % of different normal endometrium samples. The control group of OCCCs showed HNF-1-beta expression in 95 % (18/19 cases). Methylation in promoter region was detected in 13.3 % (4/30) of ECs, but not in corresponding normal tissue where available, nor in OCCCs (0/15 cases). Mutation analysis revealed truncating variant c.454C > T (p.Gln152X) in one EC and missense variant c.848C > T (p.Ala283Val) was detected in one OCCC. In conclusion, expression of HNF-1-beta was detected in various extents in all types of lesions analyzed, nevertheless its strong expression was mostly limited to clear cell carcinomas. Biological significance of genetic and epigenetic changes needs further investigation.
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http://dx.doi.org/10.1007/s12253-015-0037-2DOI Listing
July 2016

Spectral fluence of neutrons generated by radiotherapeutic linacs.

Radiat Prot Dosimetry 2015 Feb 12;163(3):373-80. Epub 2014 Jun 12.

Thomayer's Hospital, Vídeňská 80, Prague 4 CZ-140 59, Czech Republic.

Spectral fluences of neutrons generated in the heads of the radiotherapeutic linacs Varian Clinac 2100 C/D and Siemens ARTISTE were measured by means of the Bonner spheres spectrometer whose active detector of thermal neutrons was replaced by an activation detector, i.e. a tablet made of pure manganese. Measurements with different collimator settings reveal an interesting dependence of neutron fluence on the area defined by the collimator jaws. The determined neutron spectral fluences were used to derive ambient dose equivalent rate along the treatment coach. To clarify at which components of the linac neutrons are mainly created, the measurements were complemented with MCNPX calculations based on a realistic model of the Varian Clinac.
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http://dx.doi.org/10.1093/rpd/ncu192DOI Listing
February 2015

Variants of the PPARD gene and their clinicopathological significance in colorectal cancer.

PLoS One 2013 31;8(12):e83952. Epub 2013 Dec 31.

Division of Oncology, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, County Council of Östergötland, University of Linköping, Linköping, Sweden.

Background: Peroxisome proliferator-activated receptor delta (PPARD) is nuclear hormone receptor involved in colorectal cancer (CRC) differentiation and progression. The purpose of this study was to determine prevalence and spectrum of variants in the PPARD gene in CRC, and their contribution to clinicopathological endpoints.

Methods And Findings: Direct sequencing of the PPARD gene was performed in 303 primary tumors, in blood samples from 50 patients with ≥ 3 affected first-degree relatives, 50 patients with 2 affected first-degree relatives, 50 sporadic patients, 360 healthy controls, and in 6 colon cancer cell lines. Mutation analysis revealed 22 different transversions, 7 of them were novel. Three of all variants were somatic (c.548A>G, p.Y183C, c.425-9C>T, and c.628-16G>A). Two missense mutations (p.Y183C and p.R258Q) were pathogenic using in silico predictive program. Five recurrent variants were detected in/adjacent to the exons 4 (c.1-87T>C, c.1-67G>A, c.130+3G>A, and c.1-101-8C>T) and exon 7 (c.489T>C). Variant c.489C/C detected in tumors was correlated to worse differentiation (P = 0.0397).

Conclusions: We found 7 novel variants among 22 inherited or acquired PPARD variants. Somatic and/or missense variants detected in CRC patients are rare but indicate the clinical importance of the PPARD gene.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0083952PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3877104PMC
September 2014

Native red electrophoresis--a new method suitable for separation of native proteins.

Electrophoresis 2011 Dec;32(24):3597-9

Department of Biochemistry, Faculty of Science, Charles University, Hlavova, Prague, Czech Republic.

A new type of native electrophoresis was developed to separate and characterize proteins. In this modification of the native blue electrophoresis, the dye Ponceau Red S is used instead of Coomassie Brilliant Blue to impose uniform negative charge on proteins to enable their electrophoretic separation according to their relative molecular masses. As Ponceau Red S binds less tightly to proteins, in comparison with Coomassie Blue, it can be easily removed after the electrophoretic separation and a further investigation of protein properties is made possible (e.g. an enzyme detection or electroblotting). The tested proteins also kept their native properties (enzyme activity or aggregation state).
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http://dx.doi.org/10.1002/elps.201100310DOI Listing
December 2011

Native polyacrylamide electrophoresis in the presence of Ponceau Red to study oligomeric states of protein complexes.

J Sep Sci 2011 Jul 16;34(14):1692-5. Epub 2011 Jun 16.

Department of Biochemistry, Charles University, Prague, Czech Republic.

Native polyacrylamide electrophoresis in the presence of two reversible protein anionic stains (Ponceau S and Ponceau 2R) was used to study the oligomeric states of soluble proteins. A mild binding of the used protein stains to nondissociated protein oligomers imposed a charge shift on the proteins resulting into separation of protein species according to their size under physiological conditions. Adsorbed stains could be easily removed after electrophoresis by washing of polyacrylamide gel with buffer and protein complexes could be visualized either by the detection of their enzyme activity or by using a nonspecific protein stain. The specific detection of enzyme activity of glycosidases, lactate dehydrogenase, or phosphatases was shown as an example.
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http://dx.doi.org/10.1002/jssc.201000869DOI Listing
July 2011

Screening for genomic rearrangements in BRCA1 and BRCA2 genes in Czech high-risk breast/ovarian cancer patients: high proportion of population specific alterations in BRCA1 gene.

Breast Cancer Res Treat 2010 Nov 5;124(2):337-47. Epub 2010 Feb 5.

Institute of Biochemistry and Experimental Oncology, First Faculty of Medicine, Charles University in Prague, U Nemocnice 5, 128 53, Prague 2, Czech Republic.

Large genomic rearrangements (LGR) represent substantial proportion of pathogenic mutations in the BRCA1 gene, whereas the frequency of rearrangements in the BRCA2 gene is low in many populations. We screened for LGRs in BRCA1 and BRCA2 genes by multiplex ligation-dependent probe amplification (MLPA) in 521 unrelated patients negative for BRCA1/2 point mutations selected from 655 Czech high-risk breast and/or ovarian cancer patients. Besides long range PCR, a chromosome 17-specific oligonucleotide-based array comparative genomic hybridization (aCGH) was used for accurate location of deletions. We identified 14 patients carrying 8 different LGRs in BRCA1 that accounted for 12.3% of all pathogenic BRCA1 mutations. No LGRs were detected in the BRCA2 gene. In a subgroup of 239 patients from high-risk families, we found 12 LGRs (5.0%), whereas two LGRs were revealed in a subgroup of 282 non-familial cancer cases (0.7%). Five LGRs (deletion of exons 1-17, 5-10, 13-19, 18-22 and 21-24) were novel; two LGRs (deletion of exons 5-14 and 21-22) belong to the already described Czech-specific mutations; one LGR (deletion of exons 1-2) was reported from several countries. The deletions of exons 1-17 and 5-14, identified each in four families, represented Czech founder mutations. The present study indicates that screening for LGRs in BRCA1 should include patients from breast or ovarian cancer families as well as high-risk patients with non-familial cancer, in particular cases with early-onset breast or ovarian cancer. On the contrary, our analyses do not support the need to screen for LGRs in the BRCA2 gene. Implementation of chromosome-specific aCGH could markedly facilitate the design of primers for amplification and sequence analysis of junction fragments, especially in deletions overlapping gene boundaries.
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http://dx.doi.org/10.1007/s10549-010-0745-yDOI Listing
November 2010

Lack of large intragenic rearrangements in dihydropyrimidine dehydrogenase (DPYD) gene in fluoropyrimidine-treated patients with high-grade toxicity.

Cancer Chemother Pharmacol 2009 Aug 14;64(3):615-8. Epub 2009 Mar 14.

Institute of Biochemistry and Experimental Oncology, 1st Faculty of Medicine, Charles University in Prague, Prague 2, Czech Republic.

Purpose: Deficiency of dihydropyrimidine dehydrogenase (DPD) has been associated with severe fluoropyrimidines (FP) toxicity. Mutations in DPD-coding gene (DPYD) were shown to increase the risk of severe toxicity in FP-treated cancer patients. However, the majority of DPYD alterations characterized in these patients has been considered as polymorphisms and known deleterious mutations are rare and present in only limited subgroup of patients with high toxicity. Recently, the common fragile site FRA1E was mapped within DPYD locus but intragenic rearrangements in DPYD gene were not studied so far.

Methods: We performed the analysis of intragenic rearrangements of DPYD using multiplex ligation-dependent probe amplification in 68 patients with high-grade gastrointestinal and/or hematological toxicity developed at the beginning of FP treatment.

Results: We did not detect any deletion/duplication of one or more DPYD exons in analyzed patients.

Conclusions: We assume that rearrangements in DPYD gene play insignificant role in the development of serious FP-related toxicity.
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http://dx.doi.org/10.1007/s00280-009-0970-4DOI Listing
August 2009
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