Stem and Cancer Stem Cell Identities, Cellular Markers, Niche Environment and Response to Treatments to Unravel New Therapeutic Targets

Adult stem cells are a partially quiescent cell population responsible for natural cell renewal and are found in many different regions of the body, including the brain, teeth, bones, muscles, skin, and diverse epithelia, such as the epidermal or intestinal epithelium, among others [...

Testicular Germ Cell Tumours and Proprotein Convertases

Testicular Germ Cell Tumours (TGCT) are widely considered a “curable cancer” due to their exceptionally high survival rate, even if it is reduced by many years after the diagnosis due to metastases and relapses. The most common therapeutic approach to TGCTs has not changed in the last 50 years despite its multiple long-term side effects, and because it is the most common malignancy in young Caucasian men, much research is needed to better the quality of life of the many survivors. Proprotein Convertases (PC) are nine serine proteases responsible for the maturation of inactive proproteins with many diverse functions. Alterations in their expression have been associated with various diseases, including cancer and inflammation. Many of their substrates are adhesion molecules, metalloproteases and proinflammatory molecules, all of which are involved in tumour development. Inhibition of certain convertases has also been shown to slow tumour formation, demonstrating their involvement in this process. Considering the very established link between PCs and inflammation-related malignancies and the recent studies carried out into the immune microenvironment of TGCTs, the study of the involvement of PCs in testicular cancer may open up avenues for being both a biomarker for diagnosis and a therapeutic target.This research was funded by the University of the Basque Country UPV/EHU grant number GIU 20/050

Comparison of Modulation-Assisted Machining Strategies for Achieving Chip Breakage When Turning 17-4 PH Stainless Steel

Chip morphology is an intrinsic characteristic of the machining process that determines the quality of the process. When machining low machinability materials, the chips formed are usually long, continuous, and difficult to break. Due to the negative effect of the accumulation of the chip along the process, chip breakage and the correct extraction out of the machining area have become indispensable requirements. Although numerous chip-breaking methodologies have been proposed, modulation-assisted machining (MAM) is one of the most promising approaches, due to its independence from the workpiece material, tool geometry, and cutting conditions. In this work, a comparison of different modulation-assisted machining strategies, based on the modulation of the feed (F-MAM) or the depth of cut (D-MAM), were experimentally evaluated and compared to conventional turning in terms of chip morphology, surface roughness, and tool wear. Results showed that both MAM strategies enabled chip breakage and improved chip evacuation in comparison to conventional turning; however, D-MAM showed a better performance in terms of tool wear and surface roughness.This research was partially funded from the European Union’s Horizon 2020 Research and Innovation. Programme under the project InterQ (grant agreement No. 958357), and it is an initiative of the Factories-of-the-Future (FoF) Public Private Partnershi

Silencing of Sinusoidal DDR1 Reduces Murine Liver Metastasis by Colon Carcinoma

Liver metastasis depends on the collagenous microenvironment generated by hepatic sinusoidal cells (SCs). DDR1 is an atypical collagen receptor linked to tumor progression, but whether SCs express DDR1 and its implication in liver metastasis remain unknown. Freshly isolated hepatic stellate cells (HSCs), Kupffer cells (KCs), and liver sinusoidal endothelial cells (LSECs), that conform the SCs, expressed functional DDR1. HSCs expressed the largest amounts. C26 colon carcinoma secretomes increased DDR1 phosphorylation in HSCs and KCs by collagen I. Inhibition of kinase activity by DDR1-IN-1 or mRNA silencing of DDR1 reduced HSCs secretion of MMP2/9 and chemoattractant and proliferative factors for LSECs and C26 cells. DDR1-IN-1 did not modify MMP2/9 in KCs or LSECs secretomes, but decreased the enhancement of C26 migration and proliferation induced by their secretomes. Gene array showed that DDR1 silencing downregulated HSCs genes for collagens, MMPs, interleukins and chemokines. Silencing of DDR1 before tumor inoculation reduced hepatic C26 metastasis in mice. Silenced livers bore less tumor foci than controls. Metastatic foci in DDR1 silenced mice were smaller and contained an altered stroma with fewer SCs, proliferating cells, collagen and MMPs than foci in control mice. In conclusion, hepatic DDR1 promotes C26 liver metastasis and favors the pro-metastatic response of SCs to the tumor.We would like to acknowledge the following core facilities and individuals for their support: CIC bioGUNE Center for Cooperative Research in Biosciences, University of the Basque Country Animal Core Facility and SGIker Advanced Light Microscopy Core Facility. We thank Iratxe Basaldua for the in situ MMPs assay

Development, Application and Evaluation of an Active Learning Methodology for Health Science Students, Oriented towards Equity and Cultural Diversity in the Treatment and Care of Geriatric Patients

The increased aging of populations and rises in immigration have prompted the design of new methodologies and instruments for fostering the invisible care of geriatric patients among health science students in accordance with the 2030 Agenda and the SDGs. A total of 656 psychology, nursing and dentistry students participated in this study, which had a pretest–posttest design and was implemented over the course of three academic years. The intervention groups received training using an active learning methodology based on a case study involving a geriatric patient; specifically, a Maghrebi woman. The control groups were not exposed to the case study. The CCI-U questionnaire was designed ad hoc to evaluate the acquisition of invisible competences for caring for geriatric patients in accordance with their age, sex, emotional situation and ethnic origin. The questionnaire had a reliability of α = 0.63 to 0.72 and its factor solution was found to have a good fit. Students in the intervention groups scored higher than those in the control groups, with the difference being statistically significant for ethnic origin in all three undergraduate courses and all three academic years. The proper application of this active learning methodology fosters the invisible care of geriatric patients among students in accordance with the 2030 Agenda.This research project was funded by the University of the Basque Country, Office of the Pro Vice-Chancellor for Educational Innovation, grant number IKDI3-21-04, and the APC was funded by the University of the Basque Country

GeromiRs Are Downregulated in the Tumor Microenvironment during Colon Cancer Colonization of the Liver in a Murine Metastasis Model

Cancer is a phenomenon broadly related to ageing in various ways such as cell cycle deregulation, metabolic defects or telomerases dysfunction as principal processes. Although the tumor cell is the main actor in cancer progression, it is not the only element of the disease. Cells and the matrix surrounding the tumor, called the tumor microenvironment (TME), play key roles in cancer progression. Phenotypic changes of the TME are indispensable for disease progression and a few of these transformations are produced by epigenetic changes including miRNA dysregulation. In this study, we found that a specific group of miRNAs in the liver TME produced by colon cancer called geromiRs, which are miRNAs related to the ageing process, are significantly downregulated. The three principal cell types involved in the liver TME, namely, liver sinusoidal endothelial cells, hepatic stellate (Ito) cells and Kupffer cells, were isolated from a murine hepatic metastasis model, and the miRNA and gene expression profiles were studied. From the 115 geromiRs and their associated hallmarks of aging, which we compiled from the literature, 75 were represented in the used microarrays, 26 out of them were downregulated in the TME cells during colon cancer colonization of the liver, and none of them were upregulated. The histone modification hallmark of the downregulated geromiRs is significantly enriched with the geromiRs miR-15a, miR-16, miR-26a, miR-29a, miR-29b and miR-29c. We built a network of all of the geromiRs downregulated in the TME cells and their gene targets from the MirTarBase database, and we analyzed the expression of these geromiR gene targets in the TME. We found that Cercam and Spsb4, identified as prognostic markers in a few cancer types, are associated with downregulated geromiRs and are upregulated in the TME cells.This work was supported by grants from Instituto de Salud Carlos III (AC17/00012), cofounded by the European Union projects (European Regional Development Fund/European Science Foundation, Investing in your future), (ERA-Net program EracoSysMed, JTC-2 2017) and (H2020-FETOPEN, Circular Vision, Project 899417); Diputación Foral de Gipuzkoa and the Department of Economic Development and Infrastructures of the Basque Government (DFG109/20) and the Department of Economic Development and Infrastructures of the Basque Government (DFG109/Grants Health Department of the Basque Government (Spain), RIS3 call, Exp. No. 2020333039 and 2020333001. 20)

shinyCurves, a shiny web application to analyse multisource qPCR amplification data: a COVID‑19 case study

[EN]Background Quantitative, reverse transcription PCR (qRT-PCR) is currently the gold-standard for SARS-CoV-2 detection and it is also used for detection of other virus. Manual data analysis of a small number of qRT-PCR plates per day is a relatively simple task, but automated, integrative strategies are needed if a laboratory is dealing with hundreds of plates per day, as is being the case in the COVID-19 pandemic. Results Here we present shinyCurves, an online shiny-based, free software to analyze qRT-PCR amplification data from multi-plate and multi-platform formats. Our shiny application does not require any programming experience and is able to call samples Positive, Negative or Undetermined for viral infection according to a number of user-defined settings, apart from providing a complete set of melting and amplification curve plots for the visual inspection of results. Conclusions shinyCurves is a flexible, integrative and user-friendly software that speeds-up the analysis of massive qRT-PCR data from different sources, with the possibility of automatically producing and evaluating melting and amplification curve plots.This project was supported by funding from the UPV/EHU (Accion Especial "Desarrollo e implementacion del test de diagnostico para COVID-19"). The funding body did not play any roles in the study design; nor in the data collection, analysis and interpretation, or in the writing of the paper

Proprotein convertases blockage up-regulates specifically metallothioneins coding genes in human colon cancer stem cells

ABSTRACT: Despite continuous exertion made, colon cancer still represents a major health problem and its incidence continues being high worldwide. There is growing evidence in support of the cancer stem cells (CSCs) being central in the initiation of this cancer, and CSCs have been the focus of various studies for the identification of new ways of treatment. Lately, the proprotein convertases (PCs) were reported to regulate the maturation and expression of various molecules involved in the malignant phenotype of colon cancer cells, however, the identity of the molecules regulated by these serine proteases in CSCs is unknown. In this study, we used the general PCs inhibitor, the Decanoyl-RVKR-chloromethylketone (Decanoyl-RVKR-CMK) that inhibits all the PCs found in the secretory pathway, and analyzed its effect on CSCs using RNA-seq analysis. Remarkably, from the only 9 upregulated genes in the human SW620-derived sphere-forming cells, we identified 7 of the 11 human metallothioneins, all of them localized on chromosome 16, and zinc related proteins as downstream effectors of the PCs. The importance of these molecules in the regulation of cell proliferation, differentiation and chemoresistance, and their reported potential tumor suppressor role and loss in colon cancer patients associated with worse prognosis, suggests that targeting PCs in the control of the malignant phenotype of CSCs is a new potential therapeutic strategy in colon cancer

Association among University Students’ Motivation, Resilience, Perceived Competence, and Classroom Climate from the Perspective of Self-Determination Theory

Self-determination theory (SDT) suggests that motivation can interact with resilience and perceived competence. The climate-related characteristics of the classroom can influence student motivation. This study aimed to evaluate the associations between the differentiated motivation of theoretical and practical teaching, resilience, and perceived competence, considering the number of students per class and the profiles of the lecturers. A total of 789 students participated (mean age = 19.31; SD = 3.37) from Psychology, Nursing, and Education degrees from different Spanish universities. The BRS (resilience), PCNS (perceived competence), and PLOC-U (university student motivation) questionnaires were used with a new scale designed ad hoc to measure motivation in practical teaching. Student-to-class ratios and different levels of teaching experience were also recorded. A test–retest design was used to verify the stability of the measures before and after the examination of the subjects. Intrinsic motivation in practical teaching was significantly associated with resilience (r = 0.09, p < 0.03) and perceived competence (r = 0.23, p < 0.01), and in theoretical teaching, it was associated only with perceived competence (r = 20, p < 0.01). The factorial analysis of the new subscale of the PLOC-U for the measurement of motivation in practical teaching presented a good fit and reliability (α = 0.60 to 0.84) in the five factors. Test–retest analyses revealed good temporal stability. Students in small groups with more experienced lecturers scored higher on intrinsic motivation, particularly in practical classes. The stable and reliable measurement of the different types of student motivation allows their analysis and association with other variables of interest in university education, which could lead to significant improvements in teaching planning.The APC was partially funded by the University of the Basque Country and by the Biodonostia Health Research Institute

Furin Prodomain ppFurin Enhances Ca2+ Entry Through Orai and TRPC6 Channels’ Activation in Breast Cancer Cells

The intracellular calcium concentration ([Ca2+]i) modulation plays a key role in the regulation of cellular growth and survival in normal cells and failure of [Ca2+]i homeostasis is involved in tumor initiation and progression. Here we showed that inhibition of Furin by its naturally occurring inhibitor the prodomain ppFurin in the MDA-MB-231 breast cancer cells resulted in enhanced store-operated calcium entry (SOCE) and reduced the cell malignant phenotype. Expression of ppFurin in a stable manner in MDA-MB-231 and the melanoma MDA-MB-435 cell lines inhibits Furin activity as assessed by in vitro digestion assays. Accordingly, cell transfection experiments revealed that the ppFurin-expressing cells are unable to adequately process the proprotein convertase (PC) substrates vascular endothelial growth factor C (proVEGF-C) and insulin-like growth factor-1 receptor (proIGF-1R). Compared to MDA-MB-435 cells, expression of ppFurin in MDA-MB-231 and BT20 cells significantly enhanced SOCE and induced constitutive Ca2+ entry. The enhanced SOCE is impaired by inhibition of Orai channels while the constitutive Ca2+ entry is attenuated by silencing or inhibition of TRPC6 or inhibition of Orai channels. Analysis of TRPC6 activation revealed its upregulated tyrosine phosphorylation in ppFurin-expressing MDA-MB-231 cells. In addition, while ppFurin had no effect on MDA-MB-435 cell viability, in MDA-MB-231 cells ppFurin expression reduced their viability and ability to migrate and enhanced their sensitization to the apoptosis inducer hydrogen peroxide and similar results were observed in BT20 cells. These findings suggest that Furin inhibition by ppFurin may be a useful strategy to interfere with Ca2+ mobilization, leading to breast cancer cells’ malignant phenotype repression and reduction of their resistance to treatments.This research was funded by SIRIC-Brio, La Ligue Contre le Cancer and Region Nouvelle Aquitaine to A.M.K. and by PID2019-104084GB-C21 and PID2019-104084GB-C22/AEI/10.13039/501100011033, MICINN (Grant BFU2016-74932-C2) and Junta de Extremadura-Fondo Europeo de Desarrollo Regional (FEDER; Grants IB16046 and GR18061) to J.A.R. and T.S. J.J.L. is supported by a contract from Junta de Extremadura (TA18011). C.C. is supported by a contract from Junta de Extremadura—FEDER