| 1. |
- Bott, Lukas Thomas, et al.
(författare)
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Coulomb dissociation of O-16 into He-4 and C-12
- 2023
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Ingår i: NUCLEAR PHYSICS IN ASTROPHYSICS - X, NPA-X 2022. - : EDP Sciences. - 2100-014X. ; 279
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Konferensbidrag (refereegranskat)abstract
- We measured the Coulomb dissociation of O-16 into He-4 and C-12 within the FAIR Phase-0 program at GSI Helmholtzzentrum fur Schwerionenforschung Darmstadt, Germany. From this we will extract the photon dissociation cross section O-16(alpha,gamma)C-12, which is the time reversed reaction to C-12(alpha,gamma)O-16. With this indirect method, we aim to improve on the accuracy of the experimental data at lower energies than measured so far. The expected low cross section for the Coulomb dissociation reaction and close magnetic rigidity of beam and fragments demand a high precision measurement. Hence, new detector systems were built and radical changes to the (RB)-B-3 setup were necessary to cope with the high-intensity O-16 beam. All tracking detectors were designed to let the unreacted O-16 ions pass, while detecting the C-12 and He-4.
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| 2. |
- Atanasova, Diana, 1991-, et al.
(författare)
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Glycoproteomic profile of human tissue-nonspecific alkaline phosphatase expressed in osteoblasts
- 2024
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Ingår i: JBMR Plus. - : Oxford University Press. - 2473-4039. ; 8:2
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Tidskriftsartikel (refereegranskat)abstract
- Tissue-nonspecific alkaline phosphatase (TNALP) is a glycoprotein expressed by osteoblasts that promotes bone mineralization. TNALP catalyzes the hydrolysis of the mineralization inhibitor inorganic pyrophosphate and ATP to provide inorganic phosphate, thus controlling the inorganic pyrophosphate/inorganic phosphate ratio to enable the growth of hydroxyapatite crystals. N-linked glycosylation of TNALP is essential for protein stability and enzymatic activity and is responsible for the presence of different bone isoforms of TNALP associated with functional and clinical differences. The site-specific glycosylation profiles of TNALP are, however, elusive. TNALP has 5 potential N-glycosylation sites located at the asparagine (N) residues 140, 230, 271, 303, and 430. The objective of this study was to reveal the presence and structure of site-specific glycosylation in TNALP expressed in osteoblasts. Calvarial osteoblasts derived from Alpl+/− expressing SV40 Large T antigen were transfected with soluble epitope-tagged human TNALP. Purified TNALP was analyzed with a lectin microarray, matrix-assisted laser desorption/ionization-time of flight mass spectrometry, and liquid chromatography with tandem mass spectrometry. The results showed that all sites (n = 5) were fully occupied predominantly with complex-type N-glycans. High abundance of galactosylated biantennary N-glycans with various degrees of sialylation was observed on all sites, as well as glycans with no terminal galactose and sialic acid. Furthermore, all sites had core fucosylation except site N271. Modelling of TNALP, with the protein structure prediction software ColabFold, showed possible steric hindrance by the adjacent side chain of W270, which could explain the absence of core fucosylation at N271. These novel findings provide evidence for N-linked glycosylation on all 5 sites of TNALP, as well as core fucosylation on 4 out of 5 sites. We anticipate that this new knowledge can aid in the development of functional and clinical assays specific for the TNALP bone isoforms.
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| 3. |
- Escudero-Hernández, Celia, et al.
(författare)
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The Water Channel Aquaporin 8 is a Critical Regulator of Intestinal Fluid Homeostasis in Collagenous Colitis
- 2020
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Ingår i: Journal of Crohn's & Colitis. - : Oxford University Press. - 1873-9946 .- 1876-4479. ; 14:7, s. 962-973
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND AND AIMS: Diarrhoea is a common, debilitating symptom of gastrointestinal disorders. Pathomechanisms probably involve defects in trans-epithelial water transport, but the role of aquaporin [AQP] family water channels in diarrhoea-predominant diseases is unknown. We investigated the involvement of AQPs in the pathobiology of collagenous colitis [CC], which features chronic, watery diarrhoea despite overtly normal intestinal epithelial cells [IECs].METHODS: We assessed the expression of all AQP family members in mucosal samples of CC patients before and during treatment with the corticosteroid drug budesonide, steroid-refractory CC patients and healthy controls. Samples were analysed by genome-wide mRNA sequencing [RNA-seq] and quantitative real-time PCR [qPCR]. In some patients, we performed tissue microdissection followed by RNA-seq to explore the IEC-specific CC transcriptome. We determined changes in the protein levels of the lead candidates in IEC by confocal microscopy. Finally, we investigated the regulation of AQP expression by corticosteroids in model cell lines.RESULTS: Using qPCR and RNA-seq, we identified loss of AQP8 expression as a hallmark of active CC, which was reverted by budesonide treatment in steroid-responsive but not refractory patients. Consistently, decreased AQP8 mRNA and protein levels were observed in IECs of patients with active CC, and steroid drugs increased AQP8 expression in model IECs. Moreover, low APQ8 expression was strongly associated with higher stool frequency in CC patients.CONCLUSION: Down-regulation of epithelial AQP8 may impair water resorption in active CC, resulting in watery diarrhoea. Our results suggest that AQP8 is a potential drug target for the treatment of diarrhoeal disorders.
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| 4. |
- Escudero-Hernández, Celia, et al.
(författare)
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Transcriptomic profiling of collagenous colitis identifies hallmarks of non-destructive inflammatory bowel disease.
- 2021
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Ingår i: Cellular and molecular gastroenterology and hepatology. - : American Gastroenterological Association. - 2352-345X. ; 12:2, s. 665-687
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND AND AIMS: The pathophysiology of the inflammatory bowel disease collagenous colitis (CC) is poorly described. Our aim was to use RNA sequencing of mucosal samples from patients with active CC, CC in remission, refractory CC, ulcerative colitis (UC), and controls to gain insight into CC pathophysiology, identify genetic signatures linked to CC, and uncover potentially druggable disease pathways.METHODS: We performed whole transcriptome sequencing of CC samples from patients before and during treatment with the corticosteroid drug budesonide, CC steroid-refractory patients, UC patients, and healthy controls (n=9-13). Bulk mucosa and laser-captured microdissected intestinal epithelial cell (IEC) gene expression were analyzed by gene-set enrichment and gene-set variation analyses to identify significant pathways and cells, respectively, altered in CC. Leading genes and cells were validated using reverse transcription quantitative PCR and/or immunohistochemistry.RESULTS: We identified an activation of the adaptive immune response to bacteria and viruses in active CC that could be mediated by dendritic cells. Moreover, IECs display hyperproliferation and increased antigen presentation in active CC. Further analysis revealed that genes related to the immune response (DUOX2, PLA2G2A, CXCL9), DNA transcription (CTR9), protein processing (JOSD1, URI1) and ion transport (SLC9A3) remained dysregulated even after budesonide-induced remission. Budesonide-refractory CC patients fail to restore normal gene expression, and displayed a transcriptomic profile close to UC.CONCLUSIONS: Our study confirmed the implication of innate and adaptive immune responses in CC, governed by IECs and dendritic cells, respectively; and identified ongoing epithelial damage. Refractory CC could share pathomechanisms with UC.
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| 5. |
- Koch, Stefan, 1977-
(författare)
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Who controls the Wnt?
- 2019
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Ingår i: Differentiation. - : Elsevier. - 0301-4681 .- 1432-0436. ; 108
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
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| 6. |
- Molinas, Andrea, 1980-, et al.
(författare)
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The Candidate IBD Risk Gene CCNY Is Dispensable for Intestinal Epithelial Homeostasis
- 2021
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Ingår i: Cells. - : MDPI. - 2073-4409. ; 10:9
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Tidskriftsartikel (refereegranskat)abstract
- The CCNY gene, which encodes cyclin Y, has been implicated in the pathogenesis of inflammatory bowel disease (IBD). Cyclin Y promotes Wnt/β-catenin signaling and autophagy, which are critical for intestinal epithelial cell (IEC) homeostasis, and may thereby contribute to wound repair in colitis. However, whether cyclin Y has an essential function in IECs is unknown. We, therefore, investigated the epithelial injury response and mucosal regeneration in mice with conditional knock-out of Ccny in the intestinal epithelium. We observed that Ccny-deficient mice did not exhibit any differences in cell proliferation and disease activity compared to wild-type littermates in the dextran sulfate sodium (DSS) colitis model. Complementary in vitro experiments showed that loss of CCNY in model IECs did not affect Wnt signaling, cell proliferation, or autophagy. Additionally, we observed that expression of the cyclin-Y-associated cyclin-dependent kinase (CDK) 14 is exceedingly low specifically in IEC. Collectively, these results suggest that cyclin Y does not contribute to intestinal epithelial homeostasis, possibly due to low levels of specific CDKs in these cells. Thus, it is unlikely that CCNY mutations are causatively involved in IBD pathogenesis.
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| 7. |
- Moparthi, Lavanya, et al.
(författare)
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A uniform expression library for the exploration of FOX transcription factor biology
- 2020
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Ingår i: Differentiation. - : Elsevier. - 0301-4681 .- 1432-0436. ; 115, s. 30-36
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Tidskriftsartikel (refereegranskat)abstract
- Forkhead box (FOX) family transcription factors play essential roles in development, tissue homeostasis, and disease. Although the biology of several FOX proteins has been studied in depth, it is unclear to what extent these findings apply to even closely related family members, which frequently exert overlapping but non-redundant functions. To help address this question, we have generated a uniform, ready-to-use expression library of all 44 human FOX transcription factors with a convenient peptide tag for parallel screening assays. In addition, we have generated multiple universal forkhead box reporter plasmids, which can be used to monitor the transcriptional activity of most FOX proteins with high fidelity. As a proof-of-principle, we use our plasmid library to identify the DNA repair protein XRCC6/Ku70 as a selective FOX interaction partner and regulator of FOX transcriptional activity. We believe that these tools, which we make available via the Addgene plasmid repository, will considerably expedite the investigation of FOX protein biology.
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| 8. |
- Pizzolato, Giulia, 1990-
(författare)
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Molecular characterization of FOX factors and Wnt signalling interplay in human cancers
- 2023
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Wnt/β-catenin signalling, also referred to as canonical Wnt signalling, is a critical regulator of tissue homeostasis and of the differentiation of cells during development. The outcome of canonical Wnt pathway activity is defined by the regulation of target gene transcription, which ultimately determines cell identity and proliferation. How multiple proteins coordinate to modulate Wnt signalling in numerous tissues is an evolving question.Over the years, FOX transcription factors have been emerging as modulators of Wnt signalling in a variety of tissue and cell-specific contexts. Nevertheless, the function of each FOX protein in the pathway as well as their role in different pathophysiological contexts is an open matter.The overall aim of this thesis was to investigate two FOX family members, FOXB2 and FOXQ1, to uncover their roles in Wnt/β-catenin signalling. Additionally, in the last work, I aimed to investigate how the FOXQ1 oncogene is transcriptionally regulated in cancer.In the first paper, we uncovered FOXB2 as a new potent activator of Wnt signalling via the induction of agonistic Wnt ligands, particularly WNT7B. In addition, FOXB2 is induced in aggressive prostate cancer where it is associated with a neuroendocrine differentiation program and poor prognosis.In the second paper, we explored the molecular mechanisms used by the carcinoma oncogene FOXQ1 to drive Wnt signalling activation. Our results showed that FOXQ1 has a major role in tuning the Wnt transcriptional output and, in synergy with active Wnt signalling, converged on a transcriptional program linked to epithelial-to-mesenchymal transition (EMT) and cell migration, which has important implications for cancer biology.In the third paper, we reveal that p53 functions as transcriptional repressor of FOXQ1 in cancers. Loss of p53 is present in the majority of human cancers and, in synergy with activation of Wnt signalling, could boost FOXQ1 expression, thereby affecting the progression of cancer.Overall, this thesis provides a better understanding of the complexity of Wnt signalling on the molecular level and newly elucidates the function of these two FOX proteins as drivers of oncogenic Wnt pathway activation. Additionally, this new evidence highlights the importance of further in-depth investigation of FOX transcription factors in cancer biology. The relevant role of FOX proteins in the development and progression of cancer is increasingly evident, and in the long run, it will be valuable to characterize the role of FOX factors in a tissue-specific context for the development of targeted cancer therapies.
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| 9. |
- Pizzolato, Giulia, 1990-, et al.
(författare)
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The tumour suppressor p53 is a negative regulator of the carcinoma-associated transcription factor FOXQ1
- 2024
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Ingår i: Journal of Biological Chemistry. - : Elsevier. - 0021-9258 .- 1083-351X. ; 300:4
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Tidskriftsartikel (refereegranskat)abstract
- The forkhead box family transcription factor FOXQ1 is highly induced in several types of carcinomas, where it promotes epithelial-to-mesenchymal transition and tumour metastasis. The molecular mechanisms that lead to FOXQ1 deregulation in cancer are incompletely understood. Here, we used CRISPR/Cas9-based genomic locus proteomics (GLoPro) and promoter reporter constructs to discover transcriptional regulators of FOXQ1, and identified the tumour suppressor p53 as a negative regulator of FOXQ1 expression. ChIP-qPCR as well as complementary gain and loss-of-function assays in model cell lines indicated that p53 binds close to the transcription start site of the FOXQ1 promoter, and that it suppresses FOXQ1 expression in various cell types. Consistently, pharmacological activation of p53 using nutlin-3 or doxorubicin reduced FOXQ1 mRNA and protein levels in cancer cell lines harboring wild-type p53. Finally, we observed that p53 mutations are associated with increased FOXQ1 expression in human cancers. Altogether, these results suggest that loss of p53 function - a hallmark feature of many types of cancer - de-represses FOXQ1, which in turn promotes tumour progression.
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