46 research outputs found
Deletion Screening and Point Mutation Analysis in Regions of the Duchenne/Becker Muscular Dystrophy Gene
No full textDuchenne muscular dystrophy (DMD) is the commonest dystrophy with a birth incidence of one in 3000-3500 males, and approximately one third of all cases result from a new mutation. Affected males present with progressive muscle weakness and die as a result of respiratory or cardiac involvement in their late teens or early twenties. Becker muscular dystrophy (BMD) is a mild variety of the disease, and the most useful criterion for differentiating the two types is the age when patients become wheelchair bound. The gene is located on Xp21, and consists of more than 65 exons encoding a protein 3685 amino acids long, named dystrophin. About 65% of affected boys have a small submicroscopic gene deletion, heterogeneous in both the specific region and number of exons missing, that can be detected by Southern blot analysis using cDNA probes, and/or by sequence amplification with the polymerase chain reaction. Partial gene duplication accounts for the mutation in about 6% of the patients, while in the remainder point mutations are suspected An aim of this project was to screen DMD/BMD patients for deletions in exons 30 to 47 of the dystrophin gene covered by cDNA probe 5b-7, on Bglll digested DNA, and at the 3' end of the gene using the restriction enzymes Hind III and Bgl II in combination with cDNA probes 9, 10 and 11-14. Seventeen DMD/BMD patients were studied with cDNA 5b-7 and one deletion was detected having the distal end at exon 44 while the proximal end was in exon 20, in the region of cDNA probe 4-5a. Five cDNA deletions were detected having both end-points in the distal part of (lie dystrophin gene in a panel of thirty-six DMD/BMD affected males showing no deletion or duplication with the rest of the cDNA probes. Also, two cases of deletions which started in the region of cDNA 8 were identified to extend in the region of cDNA 9, with one of the two having the 3' end into the region of cDNA 10. No deletions were found with cDNA 11-14. Taking advantage of the different size of deletions detected, some of the Bgl II genomic fragments were related to Hind III genomic fragments of known orientation, and the order of the Hind III fragments in the region of cDNA 10 was rearranged. Also, the correlation between deletion and phenotype was examined and found to fit the reading-frame model proposed to explain the clinical difference in severity between DMD and BMD patients. For diagnostic purposes the detection of the molecular pathology of the disease can confirm the diagnosis of DMD/BMD in sporadic cases and offer direct accurate prenatal diagnosis in the family without the necessity for linkage analysis that requires DNA samples from key relatives. As Southern analysis takes five to ten days for results to be obtained assessment of the value and reliability of the polymerase chain reaction in performing deletion detection screening at multiple sites simultaneously was the next aim of the present study. Simultaneous amplification by PCR of exons 4, 8, 12, 17, 19, 44, 45, 48 and 51 of the dystrophin gene in 118 DMD/BMD DNA samples identified a mutation in 48. 3% of the patients and detected 86% of the deletions previously revealed by Southern analysis. Discrepancies were not found between the results obtained by the two methods of analysis. On the contrary, the problem of weak hybridisation was circumvented in two cases allowing the accurate mapping of the deletion end-points. Postnatal-detection screening was also performed in two patients and both results were confirmed by Southern analysis. The ability of PCR to amplify DNA fragments from small starting amounts of not necessarily good quality DNA permitted the amplification of DNA extracted from haematoxylin and eosin stained tissue sections, as well as the detection of a deletion in an individual whose DNA failed to produce detectable signal by Southern analysis because of degradation. Speed, sensitivity, specificity and efficiency characterise the method of multiplex amplification with the polymerase chain reaction, and make it ideal in the analysis of mutations in routine clinical practice as an initial screen to detect the molecular pathology of the disease. The final aim of this project was to scan regions of the DMD/BMD gene in affected males whose molecular pathology was still unknown, for new polymorphic sites or mutations that may account for the development of the disease and to compare and assess the different methodologies for mutational screening. (Abstract shortened by ProQuest.)
Introducing a New Historical Source on the Story, Utatsu kataki-uchi A Tōhoku Tale with an Important Episode Set at Etchū Tateyama
No full textUtatsu kataki-uchi relates a tale of murder and revenge that occurs in the village
of Utatsu, in the township of Minami-Sanriku, Miyagi Prefecture. When the parents
of siblings Ofude and Jūjirō are murdered, the children search for the killer with the
assistance of their great uncle, Jihei (a.k.a. Tarikibō). Ultimately they are able to
take revenge on the criminal, Matsuoka Mondo (a.k.a. Mikumo Danjō).
In 1995 the folklore scholar Kawashima Shūichi identified eleven recensions of
Utatsu kataki-uchi. Since then, in June of 2020 another recension sold on Yahoo’s
auction website, and more recently this author acquired an additional recension,
bringing the total number of known manuscripts of the story to thirteen. This article
introduces and transcribes the complete text of the most recently discovered recension
in order to make this source available for new research.departmental bulletin pape
Liquid Biopsy in Gastrointestinal Cancers: How Close Are We to Reaching the Clinic?
No full textAn editorial published in Cancers as part of the Special Issue Liquid Biopsy in Gastrointestinal Cancers.This research was funded by the European Union's Horizon 2020 research and innovation program under grant agreement No 857381/VISION, the Spanish Biomedical Research Network in Cancer CIBERONC (CB16/12/00446), from the Slovak Research and Development Agency (APVV-21-0197, APVV-20-0143) and TRANSCAN-2 program ERA-NET JTC 2017 "Translational research on rare cancers" within the project NExT
The Metastatic Process through the Eyes of Epigenetic Regulation: A Promising Horizon for Cancer Therapy
No full textGenetic aberrations, including chromosomal rearrangements, loss or amplification of DNA, and point mutations, are major elements of cancer development [...
The spectrum of myocardial homeostasis mechanisms in the settings of cardiac surgery procedures (Review)
No full textA machine-learning approach for pancreatic neoplasia classification based on plasma extracellular vesicles
No full textIntroduction: Pancreatic cancer (PC) is a lethal disease developing from either exocrine or endocrine cells. Efforts to assist early diagnosis focus on liquid biopsy methods, and especially on the detection of Extracellular Vesicles (EVs) secreted from cancer cells in their microenvironment and accumulated in systemic circulation. Multiple studies explore how EVs size, surface biomarkers or content can determine their unique role and function in the recipient cell’s gene expression, metabolism and behavior affecting cancer development. This study aimed to develop a machine learning-driven (ML) pipeline utilizing clinical variables and EV-based features to predict the presence of pancreatic tumors of different nature (exocrine/endocrine) in patients’ plasma compared to patients with benign lesions or age-matched non-oncological patients. Methods: All available plasma samples (N=126) and variables were collected prior to surgery. EVs were detected and characterized by flow cytometry-immunostaining. Data including size and a unique set of biomarkers (CD45, CD63 and EphA2) were combined with hematological/biochemical data and processed under two use cases, each formulated as a 3-class classification problem for patient risk stratification. The first use case aimed at classifying patients as with benign lesions or exocrine/endocrine neoplasms. The second use case aimed to distinguish patients with exocrine/endocrine neoplasms from non-oncological patients. Various ML methods were applied, including Logistic Regression, Random Forest, Support Vector Machines, and Extreme Gradient Boosting. Evaluation metrics, as area under the receiver operating characteristic curve (AUC-ROC), were computed, and Shapley values were utilized to determine features with the greatest impact on the discrimination of outcome groups. Results: Analyses identified hematological and biochemical features, among significant predictors. Models demonstrated substantial accuracy and AUC-ROC values based on plasma EVs subpopulations, which scored over 0.90 in accuracy of the Random Forest and XGBoost algorithms, presenting 0.96 +/- 0.03 accuracy in the first use case and 0.93 +/- 0.04 in the second. Discussion: By leveraging advanced analytical ML-driven approaches and integrating diverse data types, this study achieved significant accuracy, assisting patient’s risk estimation and supporting the feasibility for early detection of pancreatic cancer. Going beyond currently used biomarkers such as CEA, or CA19.9, EV-based features represent an added value offering increased diagnostic capacity. Copyright © 2025 Angelioudaki, Iosif, Kourou, Tzingounis, Kigka, Skreka, Costopoulos, Memos, Kataki, Konstadoulakis and Fotiadis
Senescence in HBV-, HCV- and NAFLD- Mediated Hepatocellular Carcinoma and Senotherapeutics: Current Evidence and Future Perspective
No full textSimple Summary Recent scientific discoveries identify cell senescence as pivotal in hepatocellular cancer (HCC) biology. Specifically, hepatitis B virus (HBV), hepatitis C virus (HCV) and non-alcoholic fatty liver disease (NAFLD) are major risk factors for HCC occurrence and it seems that cell senescence serves as a mediator. Furthermore, senescence is also implicated in HCC therapy resistance. Therefore, understanding and harnessing senescence (via senotherapeutics) seems highly important towards the discovery of new preventative and treatment strategies. Herein, we review the role of cell senescence in HBV-, HCV- and NAFLD- mediated HCC, and also explore the possible place of senotherapeutics in the management of HCC. By shining the spotlight on senescence-mediated HCC, we aim to inspire future research towards this rapidly evolving and highly promising field. Cell senescence constitutes a physiological process that serves as protection from malignant transformation of cells. However, recent scientific discoveries also identify cell senescence as pivotal in hepatocellular cancer (HCC) biology. The review herein aimed to accumulate evidence on senescence as a mediator of HCC occurrence in hepatitis B (HBV), C (HCV) virus infections, and non-alcoholic fatty liver disease (NAFLD). In HBV infection, the carcinogenic HBV X protein frequently mutates during chronic infection, and subsequently exhibits different effects on senescence. In HCV infection, senescent non-functional T-cells do not effectively clear pre-malignant hepatocytes. Furthermore, the HCV Core protein inhibits the occurrence of normal stress-induced hepatocyte senescence, allowing damaged cells to maintain their proliferative potential. In NAFLD-mediated HCC, current data point towards the gut microbiome and hepatic stellate cell senescence. Additionally, senescence contributes in the development of resistance in targeted therapies, such as sorafenib. Finally, the promising role of senotherapeutics in HCC was also explored. Overall, although we may still be at a primitive stage in fully unraveling the role of senescence in cancer, it seems that understanding and harnessing senescence may have the potential to revolutionize the way we treat hepatocellular cancer
Deliverable 3.1: Training activities
No full textTo develop and acquire scientific and technical skills, short-term staff exchanges of early stage scientists and training activities are performed within the task 3.1. This training contributes to strengthening their personal and professional development in the field of gastrointestinal cancer and nanobiotechnology. To promote the involvement of early stage researchers by enhancing the reputation, credibility, attractiveness and networking activities of BMC SAV at both international and national levels, short term staff exchanges are realized to acquire skills in various cellular, molecular and genetic techniques involved in basic and clinical research. Due to the coronavirus pandemic situation the start of this task is delayed. Various short trainings have been defined
Membranous CD44v6 is upregulated as an early event in colorectal cancer: Downregulation is associated with circulating tumor cells and poor prognosis
No full textPrevious studies have reported that CD44 variant 6 (CD44v6) and metastasis-associated protein 1 (MTA1) are contributing factors to cancer progression. The present study aimed to evaluate the expression profiles for associations with patients' demographic data, clinicopathological characteristics, the presence of partial epithelial-to-mesenchymal transition (pEMT), metastatic potential based on the presence of CK20(+) CEA(+) CXCR4(+) circulating tumor cells (CTCs) and prognosis (median follow-up, 45 months). Thus, frozen tissue samples from 31 patients with stage I-III colorectal cancer (CRC), 15 benign colorectal polyps and seven normal colorectal tissues were analyzed to detect membranous (m)CD44v6 and MTA1 expression via flow cytometry. The results demonstrated that the mCD44v6 and MTA1 expression profiles were significantly correlated (r(s)=+0.786, P<0.001). Notably, MTA1 expression was not associated with any of the clinicopathological characteristics assessed. The percentage of mCD44v6-positive cells within tumors was higher in the right-sided cancer lesions (P=0.014), suggesting that proximal and distal CRCs are distinct clinicopathological entities. Furthermore, downregulated mCD44v6 expression was significantly associated with the presence of CTCs (P=0.017). This association was stronger for pEMT (co-expression of N- and E-cadherin mRNAs) primary lesions (P=0.009). In addition, patients with CRC with low levels of mCD44v6 had unfavorable survival outcomes (P=0.037). Taken together, these results suggest that targeted analysis of membranous CD44v6 as opposed to membranous-cytoplasmic expression is important in determining the prognosis of patients with CRC. Furthermore, downregulated mCD44v6 expression in malignancies presenting CTCs reinforces the importance of tumor-stroma reciprocal influence during the metastatic process and encourages the assessment of relevant therapeutic strategies
