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  • 301.
    Defourny, Jean
    et al.
    Univ Liege, Unit Cell & Tissue Biol, GIGA Neurosci, CHU B36, B-4000 Liege, Belgium;Univ Liege, Dev Neurobiol Unit, GIGA Neurosci, CHU B36, B-4000 Liege, Belgium.
    Peuckert, Christiane
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för organismbiologi, Evolution och utvecklingsbiologi.
    Kullander, Klas
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för neurovetenskap.
    Malgrange, Brigitte
    Univ Liege, Dev Neurobiol Unit, GIGA Neurosci, CHU B36, B-4000 Liege, Belgium.
    EphA4-ADAM10 Interplay Patterns the Cochlear Sensory Epithelium through Local Disruption of Adherens Junctions2019Ingår i: iScience, E-ISSN 2589-0042 , Vol. 11, s. 246-257Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The cochlear sensory epithelium contains a functionally important triangular fluid-filled space between adjacent pillar cells referred to as the tunnel of Corti. However, the molecular mechanisms leading to local cell-cell separation during development remain elusive. Here we show that EphA4 associates with ADAM10 to promote the destruction of E-cadherin-based adhesions between adjacent pillar cells. These cells fail to separate from each other, and E-cadherin abnormally persists at the pillar cell junction in EphA4 forward-signaling-deficient mice, as well as in the presence of ADAM10 inhibitor. Using immunolabeling and an in situ proximity ligation assay, we found that EphA4 forms a complex with E-cadherin and its sheddase ADAM10, which could be activated by ephrin-B2 across the pillar cell junction to trigger the cleavage of E-cadherin. Altogether, our findings provide a new molecular insight into the regulation of adherens junctions, which might be extended to a variety of physiological or pathological processes.

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  • 302.
    Degiacomi, Giulia
    et al.
    Univ Padua, Dept Mol Med, Via Gabelli 63, I-35121 Padua, Italy..
    Personne, Yoann
    Queen Mary Univ London, London E1 2AD, England.;UCL, Div Infect & Immun, Ctr Clin Microbiol, London, England..
    Mondesert, Guillaume
    Sanofi Aventis R&D, Drug Disposit, F-69367 Lyon, France..
    Ge, Xueliang
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Struktur- och molekylärbiologi.
    Mandava, Chandra Sekhar
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Struktur- och molekylärbiologi.
    Hartkoorn, Ruben C.
    Ecole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland.;Univ Lille Nord France, Inst Pasteur Lille, Ctr Infect & Immun Lille, INSERM,CNRS,UMR 8204,U1019, Lille, France..
    Boldrin, Francesca
    Univ Padua, Dept Mol Med, Via Gabelli 63, I-35121 Padua, Italy..
    Goel, Pavitra
    Queen Mary Univ London, London E1 2AD, England..
    Peisker, Kristin
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi.
    Benjak, Andrej
    Ecole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland..
    Barrio, Maria Belen
    Sanofi Aventis R&D, Drug Disposit, F-69367 Lyon, France..
    Ventura, Marcello
    Univ Padua, Dept Mol Med, Via Gabelli 63, I-35121 Padua, Italy..
    Brown, Amanda C.
    Queen Mary Univ London, London E1 2AD, England.;Cornell Univ, Dept Microbiol & Immunol, Ithaca, NY 14853 USA..
    Leblanc, Veronique
    Sanofi Aventis R&D, Drug Disposit, F-69367 Lyon, France..
    Bauer, Armin
    Sanofi Aventis R&D, Drug Disposit, F-69367 Lyon, France..
    Sanyal, Suparna
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi.
    Cole, Stewart T.
    Ecole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland..
    Lagrange, Sophie
    Sanofi Aventis R&D, Drug Disposit, F-69367 Lyon, France..
    Parish, Tanya
    Queen Mary Univ London, London E1 2AD, England.;Infect Dis Res Inst, TB Discovery Res, Seattle, WA 98102 USA..
    Manganelli, Riccardo
    Univ Padua, Dept Mol Med, Via Gabelli 63, I-35121 Padua, Italy..
    Micrococcin P1-A bactericidal thiopeptide active against Mycobacterium tuberculosis2016Ingår i: Tuberculosis, ISSN 1472-9792, E-ISSN 1873-281X, Vol. 100, s. 95-101Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The lack of proper treatment for serious infectious diseases due to the emergence of multidrug resistance reinforces the need for the discovery of novel antibiotics. This is particularly true for tuberculosis (TB) for which 3.7% of new cases and 20% of previously treated cases are estimated to be caused by multi-drug resistant strains. In addition, in the case of TB, which claimed 1.5 million lives in 2014, the treatment of the least complicated, drug sensitive cases is lengthy and disagreeable. Therefore, new drugs with novel targets are urgently needed to control resistant Mycobacterium tuberculosis strains. In this manuscript we report the characterization of the thiopeptide micrococcin P1 as an anti-tubercular agent. Our biochemical experiments show that this antibiotic inhibits the elongation step of protein synthesis in mycobacteria. We have further identified micrococcin resistant mutations in the ribosomal protein L11 (RplK); the mutations were located in the proline loop at the N-terminus. Reintroduction of the mutations into a clean genetic background, confirmed that they conferred resistance, while introduction of the wild type RplK allele into resistant strains re-established sensitivity. We also identified a mutation in the 23S rRNA gene. These data, in good agreement with previous structural studies suggest that also in M. tuberculosis micrococcin P1 functions by binding to the cleft between the 23S rRNA and the L11 protein loop, thus interfering with the binding of elongation factors Tu and G (EF-Tu and EF-G) and inhibiting protein translocation.

  • 303.
    Deindl, Sebastian
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär systembiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Elf, Johan
    Uppsala universitet, Science for Life Laboratory, SciLifeLab. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär systembiologi.
    More Than Just Letters and Chemistry: Genomics Goes Mechanics2021Ingår i: TIBS -Trends in Biochemical Sciences. Regular ed., ISSN 0968-0004, E-ISSN 1362-4326, Vol. 46, nr 6, s. 431-432Artikel i tidskrift (Övrigt vetenskapligt)
    Abstract [en]

    Although ubiquitously thought of as a simple string of letters, DNA exhibits complex physicochemical properties. As a result, DNA can store information beyond the extensively studied explicit genetic message. The mechanical code of DNA has not been studied systematically in a genome-wide context until recent groundbreaking work by Basu et al.

  • 304.
    Dejana, Elisabetta
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Vaskulärbiologi. FIRC Inst Mol Oncol, Milan, Italy; niv Milan, Dept Oncol & Hematooncol, Milan, Italy.
    Betsholtz, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Vaskulärbiologi. Karolinska Inst, Dept Med Biochem & Biophys, Stockholm, Sweden.
    Oligodendrocytes follow blood vessel trails in the brain Brain microvasculature is a scaffold for neuroglial migration2016Ingår i: Science, ISSN 0036-8075, E-ISSN 1095-9203, Vol. 351, nr 6271, s. 341-342Artikel i tidskrift (Refereegranskat)
  • 305.
    Del Gaudio, Francesca
    et al.
    Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden..
    Liu, Dongli
    Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden.;Guangxi Med Univ Nanning, Dept Pediat, Affiliated Hosp 1, Nanning, Guangxi, Peoples R China..
    Andaloussi Mäe, Maarja
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Vaskulärbiologi.
    Braune, Eike-Benjamin
    Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden..
    Hansson, Emil M.
    Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden..
    Wang, Qing-Dong
    AstraZeneca, Biosci Cardiovasc Res & Early Dev, Cardiovasc Renal & Metab CVRM, BioPharmaceut R&D, Gothenburg, Sweden..
    Betsholtz, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Vaskulärbiologi. Karolinska Inst, Dept Med, Huddinge, Sweden..
    Lendahl, Urban
    Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden..
    Left ventricular hypertrophy and metabolic resetting in the Notch3-deficient adult mouse heart2023Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 13, nr 1, artikel-id 15022Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The heart depends on a functional vasculature for oxygenation and transport of nutrients, and it is of interest to learn how primary impairment of the vasculature can indirectly affect cardiac function and heart morphology. Notch3-deficiency causes vascular smooth muscle cell (VSMC) loss in the vasculature but the consequences for the heart remain largely elusive. Here, we demonstrate that Notch3(-/-) mice have enlarged hearts with left ventricular hypertrophy and mild fibrosis. Cardiomyocytes were hypertrophic but not hyperproliferative, and the expression of several cardiomyocyte markers, including Tnt2, Myh6, Myh7 and Actn2, was altered. Furthermore, expression of genes regulating the metabolic status of the heart was affected: both Pdk4 and Cd36 were downregulated, indicating a metabolic switch from fatty acid oxidation to glucose consumption. Notch3(-/-) mice furthermore showed lower liver lipid content. Notch3 was expressed in heart VSMC and pericytes but not in cardiomyocytes, suggesting that a perturbation of Notch signalling in VSMC and pericytes indirectly impairs the cardiomyocytes. In keeping with this, Pdgfb(ret/ret) mice, characterized by reduced numbers of VSMC and pericytes, showed left ventricular and cardiomyocyte hypertrophy. In conclusion, we demonstrate that reduced Notch3 or PDGFB signalling in vascular mural cells leads to cardiomyocyte dysfunction.

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  • 306.
    Delespaul, Lucile
    et al.
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;INSERM U1218, 229 Cours Argonne, F-33076 Bordeaux, France.;Univ Bordeaux, 146 Rue Leo Saignat, F-33000 Bordeaux, France.;Inst Canc Res, Sarcoma Mol Pathol Team, 15 Cotswold Rd, Sutton SM2 5NG, Surrey, England..
    Gélabert, Caroline
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi. INSERM U1218, 229 Cours Argonne, F-33076 Bordeaux, France.;Univ Bordeaux, 146 Rue Leo Saignat, F-33000 Bordeaux, France.;Uppsala Univ, Dept Med Biochem & Microbiol, Biomed Ctr, Sci Life Lab, Box 582, S-75123 Uppsala, Sweden..
    Lesluyes, Tom
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;INSERM U1218, 229 Cours Argonne, F-33076 Bordeaux, France.;Univ Bordeaux, 146 Rue Leo Saignat, F-33000 Bordeaux, France.;Inst Claudius Regaud, IUCT Oncopole, F-31000 Toulouse, France.;Francis Crick Inst, Canc Genom Grp, 1 Midland Rd, London NW1 1AT, England..
    Le Guellec, Sophie
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;IUCT Oncopole, Inst Claudius Regaud, Dept Pathol, F-31000 Toulouse, France..
    Perot, Gaelle
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;Inst Claudius Regaud, IUCT Oncopole, F-31000 Toulouse, France..
    Leroy, Laura
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;Inst Claudius Regaud, IUCT Oncopole, F-31000 Toulouse, France..
    Baud, Jessica
    INSERM U1218, 229 Cours Argonne, F-33076 Bordeaux, France.;Univ Bordeaux, 146 Rue Leo Saignat, F-33000 Bordeaux, France..
    Merle, Candice
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;Univ Toulouse 3 Paul Sabatier, 118 Route Narbonne, F-31062 Toulouse 9, France..
    Lartigue, Lydia
    INSERM U1218, 229 Cours Argonne, F-33076 Bordeaux, France.;Univ Bordeaux, 146 Rue Leo Saignat, F-33000 Bordeaux, France..
    Chibon, Frederic
    Canc Res Ctr Toulouse CRCT, INSERM U1037, F-31037 Toulouse, France.;IUCT Oncopole, Inst Claudius Regaud, Dept Pathol, F-31000 Toulouse, France.;ONCOSARC Team, Dept Biopathol, 2 Ave Hubert Curien,CS 53717, F-31037 Toulouse 1, France.;CRCT IUCT O, INSERM U1037, 2 Ave Hubert Curien,CS 53717, F-31037 Toulouse 1, France..
    Cell-cell fusion of mesenchymal cells with distinct differentiations triggers genomic and transcriptomic remodelling toward tumour aggressiveness2020Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 10, nr 1, artikel-id 21634Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Cell-cell fusion is a physiological process that is hijacked during oncogenesis and promotes tumour evolution. The main known impact of cell fusion is to promote the formation of metastatic hybrid cells following fusion between mobile leucocytes and proliferating tumour cells. We show here that cell fusion between immortalized myoblasts and transformed fibroblasts, through genomic instability and expression of a specific transcriptomic profile, leads to emergence of hybrid cells acquiring dissemination properties. This is associated with acquisition of clonogenic ability by fused cells. In addition, by inheriting parental properties, hybrid tumours were found to mimic the histological characteristics of a specific histotype of sarcomas: undifferentiated pleomorphic sarcomas with incomplete muscular differentiation. This finding suggests that cell fusion, as macroevolution event, favours specific sarcoma development according to the differentiation lineage of parent cells.

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  • 307.
    Deng, Xiangyi
    et al.
    Tianjin Med Univ, Key Lab Postneuroinjury Neurorepair & Regenerat C, Minist Educ & Tianjin City, Dept Neurosurg,Tianjin Neurol Inst,Gen Hosp, Tianjin, Peoples R China..
    Yang, Fan
    Tianjin Med Univ, Key Lab Postneuroinjury Neurorepair & Regenerat C, Minist Educ & Tianjin City, Dept Neurosurg,Tianjin Neurol Inst,Gen Hosp, Tianjin, Peoples R China..
    Zhang, Lei
    Shaanxi Normal Univ, Natl Engn Lab Resource Developing Endangered Chin, Minist Educ Med Plant Resource & Nat Pharmaceut C, Key Lab,Coll Life Sci, Xian, Peoples R China..
    Wang, Jianhao
    Tianjin Med Univ, Key Lab Postneuroinjury Neurorepair & Regenerat C, Minist Educ & Tianjin City, Dept Neurosurg,Tianjin Neurol Inst,Gen Hosp, Tianjin, Peoples R China..
    Liu, Boxuan
    Second Peoples Hosp Huaihua, Precis Med Ctr, Huaihua, Peoples R China..
    Liang, Wei
    Second Peoples Hosp Huaihua, Precis Med Ctr, Huaihua, Peoples R China..
    Tang, Jiefu
    Hunan Univ Med, Affiliated Hosp 1, Trauma Ctr, Huaihua, Peoples R China..
    Xie, Yuan
    Shaanxi Normal Univ, Natl Engn Lab Resource Developing Endangered Chin, Minist Educ Med Plant Resource & Nat Pharmaceut C, Key Lab,Coll Life Sci, Xian, Peoples R China..
    He, Liqun
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Vaskulärbiologi. Tianjin Med Univ, Key Lab Postneuroinjury Neurorepair & Regenerat C, Minist Educ & Tianjin City, Dept Neurosurg,Tianjin Neurol Inst,Gen Hosp, Tianjin, Peoples R China..
    ECO: An Integrated Gene Expression Omnibus for Mouse Endothelial Cells In Vivo2022Ingår i: Frontiers in Genetics, E-ISSN 1664-8021, Vol. 13, artikel-id 844544Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Endothelial cell (EC) plays critical roles in vascular physiological and pathological processes. With the development of high-throughput technologies, transcriptomics analysis of EC has increased dramatically and a large amount of informative data have been generated. The dynamic patterns of gene expression in ECs under various conditions were revealed. Unfortunately, due to the lack of bioinformatics infrastructures, reuse of these large-scale datasets is challenging for many scientists. Here, by systematic re-analyzing, integrating, and standardizing of 203 RNA sequencing samples from freshly isolated mouse ECs under 71 conditions, we constructed an integrated mouse EC gene expression omnibus (ECO). The ECO database enables one-click retrieval of endothelial expression profiles from different organs under different conditions including disease models, genetic modifications, and clinically relevant treatments in vivo. The EC expression profiles are visualized with user-friendly bar-plots. It also provides a convenient search tool for co-expressed genes. ECO facilitates endothelial research with an integrated tool and resource for transcriptome analysis. The ECO database is freely available at https://heomics. shinyapps.io/ecodb/.

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  • 308.
    Desai, Malavika Bimal
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning. Malavika Desai.
    Understanding endothelial cell distribution in Cerebral Cavernous Malformations2021Självständigt arbete på avancerad nivå (masterexamen), 40 poäng / 60 hpStudentuppsats (Examensarbete)
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  • 309.
    Di Martino, Maria Letizia
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Ek, Viktor
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Hardt, Wolf-Dietrich
    Swiss Fed Inst Technol, Inst Microbiol, Zurich, Switzerland.
    Eriksson, Jens
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Sellin, Mikael E.
    Uppsala universitet, Science for Life Laboratory, SciLifeLab. Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Barcoded Consortium Infections Resolve Cell Type-Dependent Salmonella enterica Serovar Typhimurium Entry Mechanisms2019Ingår i: mBio, ISSN 2161-2129, E-ISSN 2150-7511, Vol. 10, nr 3, artikel-id e00603-19Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Bacterial host cell invasion mechanisms depend on the bacterium's virulence factors and the properties of the target cell. The enteropathogen Salmonella enterica serovar Typhimurium (S. Tm) invades epithelial cell types in the gut mucosa and a variety of immune cell types at later infection stages. The molecular mechanism(s) of host cell entry has, however, been studied predominantly in epithelial cell lines. S. Tm uses a type three secretion system (TTSS-1) to translocate effectors into the host cell cytosol, thereby sparking actin ruffle-dependent entry. The ruffles also fuel cooperative invasion by bystander bacteria. In addition, several TTSS-1-independent entry mechanisms exist, involving alternative S. Tm virulence factors, or the passive uptake of bacteria by phagocytosis. However, it remains ill-defined how S. Tm invasion mechanisms vary between host cells. Here, we developed an internally controlled and scalable method to map S. Tm invasion mechanisms across host cell types and conditions. The method relies on host cell infections with consortia of chromosomally tagged wild-type and mutant S. Tm strains, where the abundance of each strain can be quantified by qPCR or amplicon sequencing. Using this methodology, we quantified cooccurring TTSS-1-dependent, cooperative, and TTSS-1-independent invasion events in epithelial, monocyte, and macrophage cells. We found S. Tm invasion of epithelial cells and monocytes to proceed by a similar MOI-dependent mix of TTSS-1-dependent and cooperative mechanisms. TTSS-1-independent entry was more frequent in macrophages. Still, TTSS-1-dependent invasion dominated during the first minutes of interaction also with this cell type. Finally, the combined action of the SopB/SopE/SopE2 effectors was sufficient to explain TTSS-1-dependent invasion across both epithelial and phagocytic cells. IMPORTANCE Salmonella enterica serovar Typhimurium (S. Tm) is a widespread and broad-host-spectrum enteropathogen with the capacity to invade diverse cell types. Still, the molecular basis for the host cell invasion process has largely been inferred from studies of a few selected cell lines. Our work resolves the mechanisms that Salmonellae employ to invade prototypical host cell types, i.e., human epithelial, monocyte, and macrophage cells, at a previously unattainable level of temporal and quantitative precision. This highlights efficient bacterium-driven entry into innate immune cells and uncovers a type III secretion system effector module that dominates active bacterial invasion of not only epithelial cells but also monocytes and macrophages. The results are derived from a generalizable method, where we combine barcoding of the bacterial chromosome with mixed consortium infections of cultured host cells. The application of this methodology across bacterial species and infection models will provide a scalable means to address host-pathogen interactions in diverse contexts.

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  • 310.
    Dias, Mariana Castro
    et al.
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Coisne, Caroline
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Lazarevic, Ivana
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Baden, Pascale
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Hata, Masaki
    Hyogo Coll Med, Lab Tumor Immunol & Cell Therapy, Nishinomiya, Hyogo, Japan.
    Iwamoto, Noriko
    Kobe Univ, Div Cell Biol, Grad Sch Med, Kobe, Hyogo, Japan.
    Francisco, David Miguel Ferreira
    Univ Bern, Interfac Bioinformat Unit, Bern, Switzerland;Univ Bern, Swiss Inst Bioinformat, Bern, Switzerland.
    Vanlandewijck, Michael
    AstraZeneca, Karolinska Inst, Cardio Metab Ctr, KI,AZ ICMC, Huddinge, Sweden.
    He, Liqun
    AstraZeneca, Karolinska Inst, Cardio Metab Ctr, KI,AZ ICMC, Huddinge, Sweden.
    Baier, Felix A.
    Univ Bern, Dept Biomed Res, Visceral Surg Res Lab, Bern, Switzerland.
    Stroka, Deborah
    Univ Bern, Dept Biomed Res, Visceral Surg Res Lab, Bern, Switzerland.
    Bruggmann, Remy
    Kobe Univ, Div Cell Biol, Grad Sch Med, Kobe, Hyogo, Japan;Univ Bern, Interfac Bioinformat Unit, Bern, Switzerland;Univ Bern, Swiss Inst Bioinformat, Bern, Switzerland.
    Lyck, Ruth
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Enzmann, Gaby
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Deutsch, Urban
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Betsholtz, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. AstraZeneca, Karolinska Inst, Cardio Metab Ctr, KI,AZ ICMC, Huddinge, Sweden.
    Furuse, Mikio
    Natl Inst Physiol Sci, Div Cell Struct, Okazaki, Aichi, Japan;Sch Life Sci, Dept Physiol Sci, Okazaki, Aichi, Japan.
    Tsukita, Shoichiro
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Engelhardt, Britta
    Univ Bern, Theodor Kocher Inst, Bern, Switzerland.
    Claudin-3-deficient C57BL/6J mice display intact brain barriers2019Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 9, artikel-id 203Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The tight junction protein claudin-3 has been identified as a transcriptional target of the Wnt/beta-catenin signaling pathway regulating blood-brain barrier (BBB) maturation. In neurological disorders loss of claudin-3 immunostaining is observed at the compromised BBB and blood-cerebrospinal fluid barrier (BCSFB). Although these observations support a central role of claudin-3 in regulating brain barriers' tight junction integrity, expression of claudin-3 at the brain barriers has remained a matter of debate. This prompted us to establish claudin-3-/-C57BL/6J mice to study the role of claudin-3 in brain barrier integrity in health and neuroinflammation. Bulk and single cell RNA sequencing and direct comparative qRT-PCR analysis of brain microvascular samples from WT and claudin-3-/- mice show beyond doubt that brain endothelial cells do not express claudin-3 mRNA. Detection of claudin-3 protein at the BBB in vivo and in vitro is rather due to junctional reactivity of anti-claudin-3 antibodies to an unknown antigen still detected in claudin-3-/- brain endothelium. We confirm expression and junctional localization of claudin-3 at the BCSFB of the choroid plexus. Our study clarifies that claudin-3 is not expressed at the BBB and shows that absence of claudin-3 does not impair brain barrier function during health and neuroinflammation in C57BL/6J mice.

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  • 311.
    Dierker, Tabea
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Noborn, F.
    Gothenburg Univ, Dept Clin Chem & Transfus Med, Inst Biomed, Gothenburg, Sweden..
    Hinas, Andrea
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Mikrobiologi.
    Kjellén, Lena
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Identification of novel chondroitin sulfate sulfotransferases and proteoglycan core proteins in the nematode C. elegans2017Ingår i: International journal of experimental pathology (Print), ISSN 0959-9673, E-ISSN 1365-2613, Vol. 98, nr 3, s. A4-A4Artikel i tidskrift (Övrigt vetenskapligt)
  • 312.
    Dieterich, Lothar
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Molecular Regulation of Inflammation and Angiogenesis in the Tumor Microenvironment2011Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Tumor growth and progression not only depend on properties of the malignant cells but are strongly influenced by the tumor microenvironment. The tumor stroma consists of various cell types such as inflammatory cells, endothelial cells and fibroblasts, which can either inhibit or promote tumor growth. Consequently, therapeutic targeting of the tumor stroma is increasingly recognized as an important tool to fight cancer. Two particularly important processes that contribute to the pathology of most types of tumors are angiogenesis and inflammation. In order to target these processes specifically and efficiently, it is fundamental to identify and understand the factors and signaling pathways involved.

    This thesis initially describes the multiple functions of the small heat shock protein αB-crystallin in the tumor microenvironment. αB-crystallin was first identified in a screen of proteins specifically up-regulated in endothelial cells forming vessel-like structures. We found that αB-crystallin is expressed in a subset of tumor vessels and promotes angiogenesis by inhibiting endothelial apoptosis, suggesting that targeting of αB-crystallin might inhibit angiogenesis and thereby decrease tumor growth. However, we also discovered an important role of αB-crystallin in regulation of inflammatory processes. We show that αB-crystallin increases the surface levels of E-selectin, an important leukocyte-endothelial adhesion molecule. Thereby, αB-crystallin may alter leukocyte recruitment to inflamed tissues such as the tumor stroma. In addition, we found that αB-crystallin is expressed in immature myeloid cells that accumulate in the periphery and at the tumor site during tumor development. Importantly, lack of αB-crystallin resulted in increased accumulation of immature myeloid cells, which might increase tumor associated inflammation.

    Finally, through combining laser microdissection of vessels from human tissue and microarray analysis, we identified a gene expression signature specifically associated with vessels in high grade glioma. Blood vessels in malignant glioma are highly abnormal and contribute to the pathology of the disease. Thus, knowledge about the molecular set-up of these vessels might contribute to the development of future vascular normalizing treatments.

    Delarbeten
    1. alphaB-crystallin promotes tumor angiogenesis by increasing vascular survival during tube morphogenesis
    Öppna denna publikation i ny flik eller fönster >>alphaB-crystallin promotes tumor angiogenesis by increasing vascular survival during tube morphogenesis
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    2008 (Engelska)Ingår i: Blood, ISSN 0006-4971, E-ISSN 1528-0020, Vol. 111, nr 4, s. 2015-2023Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Selective targeting of endothelial cells in tumor vessels requires delineation of key molecular events in formation and survival of blood vessels within the tumor microenvironment. To this end, proteins transiently up-regulated during vessel morphogenesis were screened for their potential as targets in antiangiogenic tumor therapy. The molecular chaperone alpha B-crystallin was identified as specifically induced with regard to expression level, modification by serine phosphorylation, and subcellular localization during tubular morphogenesis of endothelial cells. Small interfering RNA-mediated knockdown of alpha B-crystallin expression did not affect endothelial proliferation but led to attenuated tubular morphogenesis, early activation of proapoptotic caspase-3, and increased apoptosis. alpha B-crystallin was expressed in a subset of human tumor vessels but not in normal capillaries. Tumors grown in alpha B-crystallin(-/-) mice were significantly less vascularized than wild-type tumors and displayed increased areas of apoptosis/necrosis. Importantly, tumor vessels in alpha B-crystallin(-/-) mice were leaky and showed signs of caspase-3 activation and extensive apoptosis. Ultrastructural analyses showed defective vessels partially devoid of endothelial lining. These data strongly implicate alpha B-crystallin as an important regulator of tubular morphogenesis and survival of endothelial cell during tumor angiogenesis. Hereby we identify the small heat shock protein family as a novel class of anglogenic modulators.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-16117 (URN)10.1182/blood-2007-04-087841 (DOI)000253251100048 ()18063749 (PubMedID)
    Tillgänglig från: 2008-04-24 Skapad: 2008-04-24 Senast uppdaterad: 2022-01-28
    2. αB-crystallin regulates expansion of CD11b+Gr-1+ cells during tumor progression
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    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Identifikatorer
    urn:nbn:se:uu:diva-152231 (URN)
    Tillgänglig från: 2011-04-27 Skapad: 2011-04-27 Senast uppdaterad: 2011-07-01
    3. αB-crystallin influences endothelial-leukocyte interactions by increasing surface E-selectin
    Öppna denna publikation i ny flik eller fönster >>αB-crystallin influences endothelial-leukocyte interactions by increasing surface E-selectin
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    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Identifikatorer
    urn:nbn:se:uu:diva-152246 (URN)
    Tillgänglig från: 2011-04-27 Skapad: 2011-04-27 Senast uppdaterad: 2012-12-15
    4. Transcriptional profiling reveals a distinct gene expression signature of vessels in high grade human glioma
    Öppna denna publikation i ny flik eller fönster >>Transcriptional profiling reveals a distinct gene expression signature of vessels in high grade human glioma
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    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Identifikatorer
    urn:nbn:se:uu:diva-152248 (URN)
    Tillgänglig från: 2011-04-27 Skapad: 2011-04-27 Senast uppdaterad: 2011-07-01
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  • 313.
    Dieterich, Lothar C
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Schiller, Petter
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Huang, Hua
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Cancer och vaskulärbiologi.
    Wawrousek, Eric F
    National Eye Institute, National Institutes of Health.
    Loskog, Angelica
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk immunologi.
    Wanders, Alkwin
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Molekylär och morfologisk patologi.
    Moons, Lieve
    Universiteit Leuven.
    Dimberg, Anna
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Cancer och vaskulärbiologi.
    alpha B-Crystallin regulates expansion of CD11b(+)Gr-1(+) immature myeloid cells during tumor progression2013Ingår i: The FASEB Journal, ISSN 0892-6638, E-ISSN 1530-6860, Vol. 27, nr 1, s. 151-162Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The molecular chaperone αB-crystallin has emerged as a target for cancer therapy due to its expression in human tumors and its role in regulating tumor angiogenesis. αB-crystallin also reduces neuroinflammation, but its role in other inflammatory conditions has not been investigated. Here, we examined whether αB-crystallin regulates inflammation associated with tumors and ischemia. We found that CD45+ leukocyte infiltration is 3-fold increased in tumors and ischemic myocardium in αB-crystallin-deficient mice. Notably, αB-crystallin is prominently expressed in CD11b+ Gr-1+ immature myeloid cells (IMCs), known as regulators of angiogenesis and immune responses, while lymphocytes and mature granulocytes show low αB-crystallin expression. αB-Crystallin deficiency results in a 3-fold higher accumulation of CD11b+ Gr-1+ IMCs in tumors and a significant rise in CD11b+ Gr-1+ IMCs in spleen and bone marrow. Similarly, we noted a 2-fold increase in CD11b+ Gr-1+ IMCs in chronically inflamed livers in αB-crystallin-deficient mice. The effect of αB-crystallin on IMC accumulation is limited to pathological conditions, as CD11b+ Gr-1+ IMCs are not elevated in naive mice. Through ex vivo differentiation of CD11b+ Gr-1+ cells, we provide evidence that αB-crystallin regulates systemic expansion of IMCs through a cell-intrinsic mechanism. Our study suggests a key role of αB-crystallin in limiting expansion of CD11b+ Gr-1+ IMCs in diverse pathological conditions.—Dieterich, L. C., Schiller, P., Huang, H., Wawrousek, E. F., Loskog, A., Wanders, A., Moons, L., Dimberg, A. αB-Crystallin regulates expansion of CD11b+Gr-1+ immature myeloid cells during tumor progression.

  • 314.
    Digre, Andreas
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Lindskog, Cecilia
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    The Human Protein Atlas - Spatial localization of the human proteome in health and disease2021Ingår i: Protein Science, ISSN 0961-8368, E-ISSN 1469-896X, Vol. 30, nr 1, s. 218-233Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    For a complete understanding of a system's processes and each protein's role in health and disease, it is essential to study protein expression with a spatial resolution, as the exact location of proteins at tissue, cellular, or subcellular levels is tightly linked to protein function. The Human Protein Atlas (HPA) project is a large-scale initiative aiming at mapping the entire human proteome using antibody-based proteomics and integration of various other omics technologies. The publicly available knowledge resource www.proteinatlas.org is one of the world's most visited biological databases and has been extensively updated during the last few years. The current version is divided into six main sections, each focusing on particular aspects of the human proteome: (a) the Tissue Atlas showing the distribution of proteins across all major tissues and organs in the human body; (b) the Cell Atlas showing the subcellular localization of proteins in single cells; (c) the Pathology Atlas showing the impact of protein levels on survival of patients with cancer; (d) the Blood Atlas showing the expression profiles of blood cells and actively secreted proteins; (e) the Brain Atlas showing the distribution of proteins in human, mouse, and pig brain; and (f) the Metabolic Atlas showing the involvement of proteins in human metabolism. The HPA constitutes an important resource for further understanding of human biology, and the publicly available datasets hold much promise for integration with other emerging efforts focusing on single cell analyses, both at transcriptomic and proteomic level.

  • 315.
    Dijksterhuis, Jacomijn P.
    et al.
    Karolinska Inst, Sect Receptor Biol & Signaling, Deptartment Physiol & Pharmacol, S-17177 Stockholm, Sweden..
    Arthofer, Elisa
    Karolinska Inst, Sect Receptor Biol & Signaling, Deptartment Physiol & Pharmacol, S-17177 Stockholm, Sweden..
    Marinescu, Voichita D.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Nelander, Sven
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Neuroonkologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Uhlen, Mathias
    KTH Royal Inst Technol, Sci Life Lab, SE-17121 Stockholm, Sweden..
    Ponten, Frederik
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Mulder, Jan
    Karolinska Inst, Dept Neurosci, Sci Life Lab, S-17177 Stockholm, Sweden..
    Schulte, Gunnar
    Karolinska Inst, Sect Receptor Biol & Signaling, Deptartment Physiol & Pharmacol, S-17177 Stockholm, Sweden.;Masaryk Univ, Fac Sci, Inst Expt Biol, CS-61137 Brno, Czech Republic..
    High levels of WNT-5A in human glioma correlate with increased presence of tumor-associated microglia/monocytes2015Ingår i: Experimental Cell Research, ISSN 0014-4827, E-ISSN 1090-2422, Vol. 339, nr 2, s. 280-288Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Malignant gliomas are among the most severe types of cancer, and the most common primary brain tumors. Treatment options are limited and the prognosis is poor. WNT-5A, a member of the WNT family of lipoglycoproteins, plays a role in oncogenesis and tumor progression in various cancers, whereas the role of WNT-5A in glioma remains obscure. Based on the role of WNT-5A as an oncogene, its potential to regulate microglia cells and the glioma-promoting capacities of microglia cells, we hypothesize that WNT-5A has a role in regulation of immune functions in glioma. We investigated WNT-5A expression by in silico analysis of the cancer genome atlas (TCGA) transcript profiling of human glioblastoma samples and immunohistochemistry experiments of human glioma tissue microarrays (TMA). Our results reveal higher WNT-5A protein levels and mRNA expression in a subgroup of gliomas (WNT-5A(high)) compared to non-malignant control brain tissue. Furthermore, we show a significant correlation between WNT-5A in the tumor and presence of major histocompatibility complex Class II-positive microglia/monocytes. Our data pinpoint a positive correlation between WNT-5A and a proinflammatory signature in glioma. We identify increased presence of microglia/monocytes as an important aspect in the inflammatory transformation suggesting a novel role for WNT-5A in human glioma.

  • 316.
    Dimberg, Lina
    et al.
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Dimberg, Anna
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Ivarsson, Karolina
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Strömberg, Thomas
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Osterborg, Anders
    Nilsson, Kenneth
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Öberg, Fredrik
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Jernberg Wiklund, Helena
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi. Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär cellbiologi.
    Ectopic and IFN-induced expression of Fas overcomes resistance to Fas-mediated apoptosis in multiple myeloma cells.2005Ingår i: Blood, ISSN 0006-4971Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Multiple myeloma (MM) is an as yet incurable B cell malignancy. Increased survival in vitro is a hallmark of MM cells, implying that a therapeutic potential may lie in circumventing anti-apoptotic signals. We have previously reported that interferons (IFNs) sensitize MM cells to Fas/CD95-mediated apoptosis (1). In the present study, we explore the mechanism underlying this effect. In a wide screening of apoptosis-related genes, Apo2L/TRAIL and Fas were identified as IFN-targets. Sensitization to Fas-mediated apoptosis by IFNs was not affected by blocking Apo2L/TRAIL, suggesting that Apo2L/TRAIL is not a key mediator in this process. In contrast, we found that an elevated Fas expression was functionally linked to increased susceptibility to Fas-mediated apoptosis. This was further supported by the finding that IFN-treatment enhanced Fas-mediated caspase-8 activation, one of the earliest signaling events down-stream receptor activation. In addition, IFN treatment attenuated the IL-6 dependent activation of Stat3, interfering with a known survival-pathway in MM that has previously been linked with resistance to Fas-mediated apoptosis. Taken together, our results show that IFN-induced up-regulation of Fas sensitizes MM cells to Fas-mediated apoptosis and suggest that attenuation of Stat3 activation may be a potentially important event in this process.

  • 317.
    Dinic, Jelena
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Ashrafzadeh, Parham
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Parmryd, Ingela
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Actin filaments attachment at the plasma membrane in live cells cause the formation of ordered lipid domains2013Ingår i: Biochimica et Biophysica Acta - Biomembranes, ISSN 0005-2736, E-ISSN 1879-2642, Vol. 1828, nr 3, s. 1102-1111Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The relationship between ordered plasma membrane nanodomains, known as lipid rafts, and actin filaments is the focus of this study. Plasma membrane order was followed in live cells at 37°C using laurdan and di-4-ANEPPDHQ to report on lipid packing. Disrupting actin polymerisation decreased the fraction of ordered domains, which strongly argue that unstimulated cells have a basal level of ordered domains. Stabilising actin filaments had the opposite effect and increased the proportion of ordered domains. Decreasing the plasma membrane level of 4-phosphate-inositides lowers the number of attachment points for actin filaments and reduced the proportion of ordered domains. Aggregation of plasma membrane molecules, both lipid raft and non-lipid raft markers, lead to the formation of ordered domains. The increase in ordered domains was correlated with an increase in actin filaments just beneath the plasma membrane. In live cell plasma membrane blebs, which are detached from the underlying actin filaments, the fraction of ordered domains was low and GM1 could not be patched to form ordered domains. We conclude that ordered domains form when actin filaments attach to the plasma membrane. This downplays lipid-lipid interactions as the main driving force behind the formation of ordered membrane domains in vivo, giving greater prominence to membrane-intracellular filament interactions.

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  • 318.
    Do, Lan
    et al.
    Umeå universitet.
    Dahl, Christen P
    Kerje, Susanne
    Uppsala universitet, Science for Life Laboratory, SciLifeLab. Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Hansell, Peter
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi, Integrativ Fysiologi.
    Mörner, Stellan
    Lindqvist, Ulla
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Reumatologi.
    Engström-Laurent, Anna
    Larsson, Göran
    Hellman, Urban
    High Sensitivity Method to Estimate Distribution of Hyaluronan Molecular Sizes in Small Biological Samples Using Gas-Phase Electrophoretic Mobility Molecular Analysis2015Ingår i: International Journal of Cell Biology, ISSN 1687-8876, E-ISSN 1687-8884, Vol. 2015, artikel-id 938013Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Hyaluronan is a negatively charged polydisperse polysaccharide where both its size and tissue concentration play an important role in many physiological and pathological processes. The various functions of hyaluronan depend on its molecular size. Up to now, it has been difficult to study the role of hyaluronan in diseases with pathological changes in the extracellular matrix where availability is low or tissue samples are small. Difficulty to obtain large enough biopsies from human diseased tissue or tissue from animal models has also restricted the study of hyaluronan. In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan. The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs. Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before.

  • 319.
    Dolinska, Monika
    et al.
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Cai, Huan
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Mansson, Alma
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Shen, Jingyi
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Xiao, Pingnan
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Bouderlique, Thibault
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Li, Xidan
    Karolinska Univ Hosp, Karolinska Inst, Integrated Cardio Metab Ctr, Dept Med, Stockholm, Sweden..
    Leonard, Elory
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Chang, Marcus
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Gao, Yuchen
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Medina, Juan Pablo
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Kondo, Makoto
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Sandhow, Lakshmi
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Johansson, Anne-So fie
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Deneberg, Stefan
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Söderlund, Stina
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Hematologi.
    Jädersten, Martin
    Center for Hematology and Regenerative Medicine, Department of Medicine Huddinge, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
    Ungerstedt, Johanna
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Tobiasson, Magnus
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Östman, Arne
    Univ Hosp, Dept Med Sci, Div Hematol, Uppsala, Sweden. Karolinska Inst, Dept Oncol Pathol, Stockholm, Sweden..
    Mustjoki, Satu
    Univ Helsinki, Helsinki Univ Hosp, Hematol Res Unit Helsinki, Comprehens Canc Ctr, Helsinki, Finland.;Univ Helsinki, Dept Clin Chem & Hematol, Translat Immunol Res Program, Helsinki, Finland.;iCAN Digital Precis Canc Med Flagship, Helsinki, Finland..
    Stenke, Leif
    Karolinska Univ Hosp, Div Hematol, Stockholm, Sweden.;Karolinska Inst, Dept Med Solna, Stockholm, Sweden..
    Le Blanc, Katarina
    Karolinska Univ Hosp, Div Clin Immunol & Transfus Med, Stockholm, Sweden..
    Hellström-Lindberg, Eva
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Lehmann, Sören
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Hematologi. Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden..
    Ekblom, Marja
    Lund Univ, Div Mol Hematol, Lund, Sweden..
    Olsson-Strömberg, Ulla
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Hematologi.
    Sigvarosson, Mikael
    Lund Univ, Div Mol Hematol, Lund, Sweden.;Linköping Univ, Dept Clin & Expt Med, Linköping, Sweden..
    Qian, Hong
    Karolinska Inst, Ctr Hematol & Regenerat Med, Dept Med Huddinge, Stockholm, Sweden.;Karolinska Inst, Ctr Hematol & Regenerat Med HERM, Dept Med Huddinge, Neo Floor 7, S-14186 Stockholm, Sweden..
    Characterization of the bone marrow niche in patients with chronic myeloid leukemia identifies CXCL14 as a new therapeutic option2023Ingår i: Blood, ISSN 0006-4971, E-ISSN 1528-0020, Vol. 142, nr 1, s. 73-89Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Although tyrosine kinase inhibitors (TKIs) are effective in treating chronic myeloid leukemia (CML), they often fail to eradicate the leukemia-initiating stem cells (LSCs), causing disease persistence and relapse. Evidence indicates that LSC persistence may be because of bone marrow (BM) niche protection; however, little is known about the underlying mechanisms. Herein, we molecularly and functionally characterize BM niches in patients with CML at diagnosis and reveal the altered niche composition and function in these patients. Long -term culture initiating cell assay showed that the mesenchymal stem cells from patients with CML displayed an enhanced supporting capacity for normal and CML BM CD34+CD38- cells. Molecularly, RNA sequencing detected dysregulated cytokine and growth factor expression in the BM cellular niches of patients with CML. Among them, CXCL14 was lost in the BM cellular niches in contrast to its expression in healthy BM. Restoring CXCL14 significantly inhibited CML LSC maintenance and enhanced their response to imatinib in vitro, and CML engraftment in vivo in NSG-SGM3 mice. Importantly, CXCL14 treatment dramatically inhibited CML engraftment in patient-derived xenografted NSG-SGM3 mice, even to a greater degree than imatinib, and this inhibition persisted in patients with suboptimal TKI response. Mechanistically, CXCL14 upregulated inflammatory cytokine signaling but downregulated mTOR signaling and oxidative phosphorylation in CML LSCs. Together, we have discovered a suppressive role of CXCL14 in CML LSC growth. CXCL14 might offer a treatment option targeting CML LSCs.

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  • 320. Dong, Chuanjiang
    et al.
    Chen, Wei
    Zou, Lili
    Liu, Binbin
    Deng, Kaihong
    Guo, Dingrui
    Wang, Peng
    Chen, Hao
    Wang, Helen
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Wang, Jun
    The Assessment on Synergistic Activity of Ebselen and Silver Ion Against Yersinia pseudotuberculosis2022Ingår i: Frontiers in Microbiology, E-ISSN 1664-302X, Vol. 13Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Yersinia pseudotuberculosis is a foodborne zoonotic bacterium that is pathogenic to guinea pigs, rabbits, and mice. It also causes pseudotuberculosis in humans. However, it still lacked the scientific basis for control. Here, we found out that Ebselen (EbSe) exhibited synergistic antibacterial activity with silver nitrate (Ag+) against Y. pseudotuberculosis YpIII strain with high efficacy in vitro using UV-visible light absorption spectrum, 5,5’-dithiobis-(2-nitrobenzoic acid), laser scanning confocal microscope, flow cytometry, transmission electron microscopy and Western blotting assays. The depletion of total glutathione (GSH) amount and inhibition of thioredoxin reductase (TrxR) activity in thiol-dependent redox system revealed the destructiveness of EbSe-Ag+-caused intracellular oxidative stress. Furthermore, a YpIII-caused mice gastroenteritis model was constructed. EbSe-Ag+ significantly reduced bacterial loads with low toxicity. It also down-regulated the expression levels of interferon (IL)-1β and tumor necrosis factor-α, up-regulated the expression level of IL-10 on-site. All the in vivo results demonstrated the antibacterial activity and immune-modulatory property of EbSe-Ag+. Collectively, these results provided academic fundament for further analysis and development of EbSe-Ag+ as the antibacterial agents for pseudotuberculosis control.

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    fulltext
  • 321.
    Dou, Zelong
    et al.
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden..
    Muder, Daniel
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper, Ortopedi. Falu Lasarett, Dept Orthoped, Lasarettsvagen 10, SE-79182 Falun, Sweden..
    Baroncelli, Marta
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden..
    Bendre, Ameya
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden..
    Gkourogianni, Alexandra
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden..
    Ottosson, Lars
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden..
    Vedung, Torbjörn
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper, Ortopedi. Elisabeth Hosp, Aleris Hlthcare, Uppsala, Sweden..
    Nilsson, Ola
    Karolinska Inst, Div Pediat Endocrinol, Stockholm, Sweden.;Karolinska Inst & Univ Hosp, Ctr Mol Med L8 01, Dept Womens & Childrens Hlth, SE-17176 Stockholm, Sweden.;Univ Hosp, Stockholm, Sweden.;Örebro Univ & Univ Hosp, Sch Med Sci, Örebro, Sweden..
    Rat perichondrium transplanted to articular cartilage defects forms articular-like, hyaline cartilage2021Ingår i: Bone, ISSN 8756-3282, E-ISSN 1873-2763, Vol. 151, artikel-id 116035Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Objective: Perichondrium autotransplants have been used to reconstruct articular surfaces destroyed by infection or trauma. However, the role of the transplanted perichondrium in the healing of resurfaced joints has not been investigated.

    Design: Perichondrial and periosteal tissues were harvested from rats hemizygous for a ubiquitously expressed enhanced green fluorescent protein (EGFP) transgene and transplanted into full-thickness articular cartilage defects at the trochlear groove of distal femur in wild-type littermates. As an additional control, cartilage defects were left without a transplant (no transplant control). Distal femurs were collected 3, 14, 56, 112 days after surgery.

    Results: Tracing of transplanted cells showed that both perichondrium and periosteum transplant-derived cells made up the large majority of the cells in the regenerated joint surfaces. Perichondrium transplants contained SOX9 positive cells and with time differentiated into a hyaline cartilage that expanded and filled out the defects with Col2a1-positive and Col1a1-negative chondrocytes and a matrix rich in proteoglycans. At later timepoints the cartilaginous perichondrium transplants were actively remodeled into bone at the transplant-bone interface and at post-surgery day 112 EGFP-positive perichondrium cells at the articular surface were positive for Prg4. Periosteum transplants initially lacked SOX9 expression and despite a transient increase in SOX9 expression and chondrogenic differentiation, remained Col1a1 positive, and were continuously thinning as periosteum-derived cells were incorporated into the subchondral compartment.

    Conclusions: Perichondrium and periosteum transplanted to articular cartilage defects did not just stimulate regeneration but were themselves transformed into cartilaginous articular surfaces. Perichondrium transplants developed into an articular-like, hyaline cartilage, whereas periosteum transplants appeared to produce a less resilient fibro-cartilage.

  • 322.
    Dourado, Daniel F. A. R.
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Beräkningsbiologi och bioinformatik.
    Flores, Samuel Coulbourn
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Beräkningsbiologi och bioinformatik.
    Modeling and fitting protein-protein complexes to predict change of binding energy2016Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 6, artikel-id 25406Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    It is possible to accurately and economically predict change in protein-protein interaction energy upon mutation (Delta Delta G), when a high-resolution structure of the complex is available. This is of growing usefulness for design of high-affinity or otherwise modified binding proteins for therapeutic, diagnostic, industrial, and basic science applications. Recently the field has begun to pursue Delta Delta G prediction for homology modeled complexes, but so far this has worked mostly for cases of high sequence identity. If the interacting proteins have been crystallized in free (uncomplexed) form, in a majority of cases it is possible to find a structurally similar complex which can be used as the basis for template-based modeling. We describe how to use MMB to create such models, and then use them to predict Delta Delta G, using a dataset consisting of free target structures, co-crystallized template complexes with sequence identify with respect to the targets as low as 44%, and experimental Delta Delta G measurements. We obtain similar results by fitting to a low-resolution Cryo-EM density map. Results suggest that other structural constraints may lead to a similar outcome, making the method even more broadly applicable.

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  • 323.
    Dragomir, Anca
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Approaches to Pharmacological Treatment and Gene Therapy of Cystic Fibrosis2004Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Cystic fibrosis (CF) is the most common lethal genetic disease in the white population. It is due to mutations in the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR), a protein that functions mainly as a cAMP-activated chloride channel. The disease impairs ion and water transport in epithelia-lined organs such as airways, digestive tract, reproductive epithelium and sweat glands. At present the only therapy is symptomatic and development of curative treatment depends on uncovering the links between the defective CFTR and the disease, as well as on improving end-point measurements.

    A method has been established for studying ion transport in an easily accessible cell type (nasal epithelial cells) from normal and cystic fibrosis patients by X-ray microanalysis. This method represents a rather simple and direct way of measuring simultaneously several chemical elements of biological interest.

    Studies of chloride transport by means of a fluorescent indicator (MQAE) in nasal epithelial cells from CF patients showed that the phenotype cannot exclusively be explained by the CFTR activity in patients with severe genotype.

    A common Portuguese CFTR mutation (A561E) causes protein mislocalization in the endoplasmic reticulum similar to the most common CF mutation (ΔF508) and thus it should be possible to treat it with the same pharmacological strategies.

    Chronic treatment of CF airway epithelial cells with nanomolar concentrations of colchicine increased the chloride efflux via chloride channels other than CFTR, strengthening the notion that colchicine could be beneficial to CF patients.

    Successful in vitro transfection of CF airway epithelial cells with cationic vectors was possible with short incubation times. Heparin added at the end of the transfection incubation time could help to maintain the viability of the cells, without interfering with the transfection efficiency. It seems possible that heparin could be an adjuvant for non-viral mediated gene therapy.

    Delarbeten
    1. Assessment of chloride secretion in human nasal epithelial cells by X-ray microanalysis.
    Öppna denna publikation i ny flik eller fönster >>Assessment of chloride secretion in human nasal epithelial cells by X-ray microanalysis.
    Visa övriga...
    2001 Ingår i: Journal of Microscopy, Vol. 203, nr 3, s. 277-284Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-91153 (URN)
    Tillgänglig från: 2003-12-17 Skapad: 2003-12-17Bibliografiskt granskad
    2. Cystic fibrosis transmembrane conductance regulator (CFTR) activity in nasal epithelial cells from cystic fibrosis patients with severe genotypes.
    Öppna denna publikation i ny flik eller fönster >>Cystic fibrosis transmembrane conductance regulator (CFTR) activity in nasal epithelial cells from cystic fibrosis patients with severe genotypes.
    2002 Ingår i: Clinical Science, Vol. 103, s. 417-424Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-91154 (URN)
    Tillgänglig från: 2003-12-17 Skapad: 2003-12-17Bibliografiskt granskad
    3. Unusually common cystic fibrosis mutation in Portugal encodes a misprocessed protein.
    Öppna denna publikation i ny flik eller fönster >>Unusually common cystic fibrosis mutation in Portugal encodes a misprocessed protein.
    Visa övriga...
    2003 Ingår i: Biochemical and Biophysical Research Communications, Vol. 311, s. 665-671Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-91155 (URN)
    Tillgänglig från: 2003-12-17 Skapad: 2003-12-17Bibliografiskt granskad
    4. Colchicine increases the chloride efflux in airway epithelial cell lines.
    Öppna denna publikation i ny flik eller fönster >>Colchicine increases the chloride efflux in airway epithelial cell lines.
    Artikel i tidskrift (Refereegranskat) Submitted
    Identifikatorer
    urn:nbn:se:uu:diva-91156 (URN)
    Tillgänglig från: 2003-12-17 Skapad: 2003-12-17Bibliografiskt granskad
    5. Heparin can improve the viability of transfected cystic fibrosis cell lines in vitro.
    Öppna denna publikation i ny flik eller fönster >>Heparin can improve the viability of transfected cystic fibrosis cell lines in vitro.
    Artikel i tidskrift (Refereegranskat) Submitted
    Identifikatorer
    urn:nbn:se:uu:diva-91157 (URN)
    Tillgänglig från: 2003-12-17 Skapad: 2003-12-17Bibliografiskt granskad
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  • 324. Dransart, Estelle
    et al.
    Di Cicco, Aurélie
    El Marjou, Ahmed
    Lévy, Daniel
    Johansson, Staffan
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Johannes, Ludger
    Shafaq-Zadah, Massiullah
    Solubilization and Purification of α5β1 Integrin from Rat Liver for Reconstitution into Nanodiscs2022Ingår i: Methods in Molecular Biology, Vol. 2507. / [ed] Isabelle Mus-Veteau, Springer, 2022, s. 1-18Kapitel i bok, del av antologi (Refereegranskat)
  • 325.
    Dryselius, Staffan
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Studies of individual pancreatic -cells: Electrophysiological analysis of rhythmic behaviour and development of new techniques1999Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    The insulin concentration in blood varies periodically, which is believed to prevent down-regulation of the hormone receptors. Loss of the regular insulin oscillations is considered to be an early sign of developing diabetes. The insulin variations are due to pulsatile release of insulin from the pancreatic islets and their β-cells, which occurs in synchrony with slow large amplitude oscillations of the cytoplasmic Ca2+ concentration ([Ca2+]i). It was now investigated whether oscillations in metabolism may drive those in [Ca2+]i or if metabolism follows [Ca2+]i. Using the ATP concentration as an indicator of metabolism, the activity of ATP-regulated K+ (KATP) channels was analysed in the cell-attached configuration of the patch clamp technique. At 0-3 mM glucose, a situation with low and stable [Ca2+]i, there were regular slow fluctuations in KATP channel open-time. These variations, with a frequency similar to that of the slow [Ca2+]i oscillations, provide strong evidence that metabolic oscillations are primary events. Ca2+ entry through voltage-activated channels was evaluated as a mechanism for generating the slow [Ca2+]i oscillations. Using Sr2+ and Mn2+ as analogues for Ca2+, glucose stimulation and depolarisation with K+ were found to enhance the influx. Moreover, the [Ca2+]i oscillations were almost perfectly parallel with slow bursts of action currents. Confirming a central role of voltage-dependent Ca2+ entry all these effects were inhibited by the specific Ca2+ channel antagonist nifedipine. During the bursts occasional pronounced [Ca2+]i spikes, due to intracellular release of the ion, temporarily arrested the action currents. It is discussed how these spikes may generate a hyperpolarising current, explaining the generation of the fast oscillatory pattern observed in pancreatic islets. The wavelet transform was found to be an excellent tool for reducing noise in experimental data and for highlighting recurrent electrophysiological patterns. An improved U-tube technique is presented, allowing change of medium around single cells within 60 msec. An engraving tool, producing 1.5 µm broad lines, was constructed and used to label light microscope object and cover glasses with micro-text.

  • 326.
    Dupret, Vincent
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för organismbiologi, Evolution och utvecklingsbiologi.
    Sanchez, Sophie
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för organismbiologi, Evolution och utvecklingsbiologi.
    Goujet, Daniel
    Muséum National D'Histoire Naturelle, Paris, France.
    Tafforeau, Paul
    European Synchrotron Radiation Facility, Grenoble, France.
    Ahlberg, Per Erik
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för organismbiologi, Evolution och utvecklingsbiologi.
    Bone vascularization and growth in placoderms (Vertebrata): The example of the premedian plate of Romundina stellina Ørvig, 19752010Ingår i: Comptes rendus. Palevol, ISSN 1631-0683, E-ISSN 1777-571X, Vol. 9, nr 6-7, s. 369-375Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The Placodermi (armored jawed fishes), which appeared during the Lower Silurian and disappeared without leading any descendants at the end of the Famennian (Latest Devonian), have the highest diversity of known Devonian vertebrate groups. As phylogenetically basal gnathostomes (jawed vertebrates), they are potentially informative about primitive jawed vertebrate anatomy and origins. Until recently, the study of their internal or histological structures has required destructive methods such as sectioning or serial grinding. Recent advances in tomography and imaging technologies, especially through the increasing use of synchrotron phase contrast imaging for the study of fossils, allow us to reveal the inner structures of the fossil nondestructively and with unprecedented three-dimensional level of detail. Here, we present for the first time the prerostral anatomy of the small acanthothoracid Romundina stellina, one of the earliest and most basal placoderms. Phase contrast imaging allows us to reconstruct the vascularization and nerve canals of the premedian plate and adjacent parts of the skeleton three-dimensionally in great detail, providing important clues to the growth modes and biology of the animal.

  • 327.
    Dyachok, Oleg
    et al.
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Gylfe, Erik
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Ca2+-induced Ca2+ Release via Inositol 1,4,5-trisphosphate Receptors Is Amplified by Protein Kinase A and Triggers Exocytosis in Pancreatic β-Cells2004Ingår i: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 279, nr 44, s. 45455-45461Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Hormones, such as glucagon and glucagon-like peptide-1, potently amplify nutrient stimulated insulin secretion by raising cAMP. We have studied how cAMP affects Ca2+-induced Ca2+ release (CICR) in pancreatic β-cells from mice and rats and the role of CICR in secretion. CICR was observed as pronounced Ca2+ spikes on top of glucose- or depolarization-dependent rise of the cytoplasmic Ca2+ concentration ([Ca2+]i). cAMP-elevating agents strongly promoted CICR. This effect involved sensitization of the receptors underlying CICR, because many cells exhibited the characteristic Ca2+ spiking at low or even in the absence of depolarization-dependent elevation of [Ca2+]i. The cAMP effect was mimicked by a specific activator of protein kinase A in cells unresponsive to activators of cAMP-regulated guanine nucleotide exchange factor. Ryanodine pretreatment, which abolishes CICR mediated by ryanodine receptors, did not prevent CICR. Moreover, a high concentration of caffeine, known to activate ryanodine receptors independently of Ca2+, failed to mobilize intracellular Ca2+. On the contrary, a high caffeine concentration abolished CICR by interfering with inositol 1,4,5-trisphosphate receptors (IP3Rs). Therefore, the cell-permeable IP3R antagonist 2-aminoethoxydiphenyl borate blocked the cAMP-promoted CICR. Individual CICR events in pancreatic β-cells were followed by [Ca2+]i spikes in neighboring human erythroleukemia cells, used to report secretory events in the β-cells. The results indicate that protein kinase A-mediated promotion of CICR via IP3Rs is part of the mechanism by which cAMP amplifies insulin release.

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    FULLTEXT02
  • 328.
    Dyachok, Oleg
    et al.
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Gylfe, Erik
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Store-operated influx of Ca2+ in the pancreatic β-cells exhibits graded dependence on the filling of the endoplasmic reticulum2001Ingår i: Journal of Cell Science, ISSN 0021-9533, E-ISSN 1477-9137, Vol. 114, nr Pt 11, s. 2179-2186Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The store-operated pathway for Ca2+ entry was studied in individual mouse pancreatic β-cells by measuring the cytoplasmic concentrations of Ca2+ ([Ca2+]i) and Mn2+ ([Mn2+]i) with the fluorescent indicator fura-2. Influx through the store-operated pathway was initially shut off by pre-exposure to 20 mM glucose, which maximally stimulates intracellular Ca2+ sequestration. To avoid interference with voltage-dependent Ca2+ entry the cells were hyperpolarized with diazoxide and the channel blocker methoxyverapamil was present. Activation of the store-operated pathway in response to Ca2+ depletion of the endoplasmic reticulum was estimated from the sustained elevation of [Ca2+]i or from the rate of increase in [Mn2+]i due to influx of these extracellular ions. Increasing concentrations of the inositol 1,4,5-trisphosphate-generating agonist carbachol or the sarco(endo)plasmatic reticulum Ca2+-ATPase inhibitor cyclopiazonic acid (CPA) cause gradual activation of the store-operated pathway. In addition, the carbachol- and CPA-induced influx of Mn2+ depended on store filling in a graded manner. The store-operated influx of Ca2+/Mn2+ was inhibited by Gd3+ and 2-aminoethoxydiphenyl borate but neither of these agents discriminated between store-operated and voltage-dependent entry. The finely tuned regulation of the store-operated mechanisms in the β-cell has direct implications for the control of membrane potential and insulin secretion.

    Ladda ner fulltext (pdf)
    FULLTEXT01
  • 329.
    Dyachok, Oleg
    et al.
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Tufveson, Gunnar
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper, Transplantationskirurgi.
    Gylfe, Erik
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Ca2+-induced Ca2+ release by activation of inositol 1,4,5-trisphosphate receptors in primary pancreatic β-cells2004Ingår i: Cell Calcium, ISSN 0143-4160, E-ISSN 1532-1991, Vol. 36, nr 1, s. 1-9Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The effect of sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) inhibition on the cytoplasmic Ca2+ concentration ([Ca2+]i) was studied in primary insulin-releasing pancreatic β-cells isolated from mice, rats and human subjects as well as in clonal rat insulinoma INS-1 cells. In Ca2+-deficient medium the individual primary β-cells reacted to the SERCA inhibitor cyclopiazonic acid (CPA) with a slow rise of [Ca2+]i followed by an explosive transient elevation. The [Ca2+]i transients were preferentially observed at low intracellular concentrations of the Ca2+ indicator fura-2 and were unaffected by pre-treatment with 100 μM ryanodine. Whereas 20 mM caffeine had no effect on basal [Ca2+]i or the slow rise in response to CPA, it completely prevented the CPA-induced [Ca2+]i transients as well as inositol 1,4,5-trisphosphate-mediated [Ca2+]i transients in response to carbachol. In striking contrast to the primary β-cells, caffeine readily mobilized intracellular Ca2+ in INS-1 cells under identical conditions, and such mobilization was prevented by ryanodine pre-treatment. The results indicate that leakage of Ca2+ from the endoplasmic reticulum after SERCA inhibition is feedback-accelerated by Ca2+-induced Ca2+ release (CICR). In primary pancreatic β-cells this CICR is due to activation of inositol 1,4,5-trisphosphate receptors. CICR by ryanodine receptor activation may be restricted to clonal β-cells.

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    FULLTEXT02
  • 330.
    Eberhardson, Michael
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    The interplay of ions in the stimulation of the pancreatic -cell1999Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Glucose stimulation of insulin release is mediated by depolarisation of the pancreatic β-cells with accompanying entry of Ca2+ through voltage-dependent channels. An important feature of the glucose-induced depolarisation is its rhythmicity causing oscillations of the cytoplasmic Ca2+ concentration ([Ca2+]i), which trigger pulsatile release of insulin. In addition to inducing slow [Ca2+]i oscillations (0.2-0.5/min), glucose is known to be a cofactor for brief transients of 10-20 sec duration due to intracellular mobilisation of Ca2+. In the present study microfluorometric technique was used for elucidating how various ion permeabilities interact in the generation of Ca2+ signals in individual β-cells. Increased Ca2+ entry or inhibition of K+ channels promoted the glucose-induced slow oscillations of [Ca2+]i. Studying how inhibition of K+ channels contributed to the generation of slow oscillations, it was discovered that tetraehylammonium+, quinine, and Cs+ precipitated pronounced Ca2+ transients in β-cells stimulated with glucose or tolbutamide. Similar transients were obtained when the entry of Na+ was stimulated with the Na+ channel agonist veratridine. The transients differed from those previously described in reflecting voltage-dependent entry of Ca2+ instead of mobilisation of intracellular Ca2+ stores. The steady-state concentration of Cl- in unstimulated β-cells was found to be 34 mM, indicating that Cl- is accumulated against its electrochemical gradient. Further studies showed that glucose increases the Cl- permeability, an effect which is supposed to contribute to the depolarising action of the sugar. Evidence was provided that transmembrane Cl- fluxes are important for the generation of the slow oscillations as well as for the fast transients of [Ca2+]i, which both depend on entry of Ca2+ through voltage-dependent channels.

  • 331. Eckerle, S.
    et al.
    Brune, V.
    Döring, C.
    Tiacci, E.
    Bohle, V.
    Sundström, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Kodet, R.
    Paulli, M.
    Falini, B.
    Klapper, W.
    Chaubert, A. B.
    Willenbrock, K.
    Metzler, D.
    Bräuninger, A.
    Küppers, Ralf
    Hansmann, M-L.
    Gene expression profiling of isolated tumour cells from anaplastic large cell lymphomas: insights into its cellular origin, pathogenesis and relation to Hodgkin lymphoma2009Ingår i: Leukemia, ISSN 0887-6924, E-ISSN 1476-5551, Vol. 23, nr 11, s. 2129-2138Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Anaplastic large cell lymphoma (ALCL) is a main type of T-cell lymphomas and comprises three distinct entities: systemic anaplastic lymphoma kinase (ALK) positive, systemic ALK(-) and cutaneous ALK(-) ALCL (cALCL). Little is known about their pathogenesis and their cellular origin, and morphological and immunophenotypical overlap exists between ALK(-) ALCL and classical Hodgkin lymphoma (cHL). We conducted gene expression profiling of microdissected lymphoma cells of five ALK(+) and four ALK(-) systemic ALCL, seven cALCL and sixteen cHL, and of eight subsets of normal T and NK cells. The analysis supports a derivation of ALCL from activated T cells, but the lymphoma cells acquired a gene expression pattern hampering an assignment to a CD4(+), CD8(+) or CD30(+) T-cell origin. Indeed, ALCL display a down-modulation of many T-cell characteristic molecules. All ALCL types show significant expression of NFkappaB target genes and upregulation of genes involved in oncogenesis (e.g. EZH2). Surprisingly, few genes are differentially expressed between systemic and cALCL despite their different clinical behaviour, and between ALK(-) ALCL and cHL despite their different cellular origin. ALK(+) ALCL are characterized by expression of genes regulated by pathways constitutively activated by ALK. This study provides multiple novel insights into the molecular biology and pathogenesis of ALCL.

  • 332.
    Edlund, Karolina
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
    Microdissection of well defined cell populations for RNA isolation in the analysis of normal human skin and basal cell carcinoma2005Självständigt arbete på grundnivå (kandidatexamen), 10 poäng / 15 hpStudentuppsats
    Abstract [en]

    The human skin provides us with an excellent protective barrier and possesses a remarkable ability of constant renewal. Basal cell carcinoma is the most common type of skin cancer. The aim of this project was to verify results from an earlier study investigating the molecular differences between basal cell carcinoma (BCC) and basal cells of normal human epidermis. In that study microdissection of cell populations from BCC and basal cells of normal epidermis respectively was performed in five cases of confirmed BCC. Following RNA extraction and amplification, a gene expression analysis was performed using a 46 k human cDNA microarray. Comparison of expression profiles showed a differential expression of approximately 300 genes in BCC. An upregulation of signaling pathways previously known to be of importance in BCC development could be observed, as well as a downregulation of differentiation markers, MHC class II molecules, and proteins active in scavenging of oxygen radicals. We wanted to confirm these findings for a number of selected genes, using real time PCR. The focal point of this project was microdissection of cells from BCC and subsequent isolation of RNA. Microdissection based methods offer a possibility of selecting well defined cell populations for further analysis by using a focused laser beam. Initially tests in order to optimize the method were also performed, concerning the dehydration process and choice of slides used in microdissection. Isolation of RNA may, as we experienced, be associated with problems due to destruction of RNA by degrading enzymes.

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  • 333.
    Edlund, Karolina
    et al.
    TU Dortmund Univ, Leibniz Res Ctr Working Environm & Human Factors, Dortmund, Germany.
    Madjar, Katrin
    TU Dortmund Univ, Dept Stat, Dortmund, Germany.
    Mattsson, Johanna Sofia Margareta
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Djureinovic, Dijana
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Lindskog, Cecilia
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Brunnström, Hans
    Lund Univ, Dept Clin Sci, Div Oncol & Pathol, Lund, Sweden.
    Koyi, Hirsh
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Centrum för klinisk forskning, Gävleborg. Gävle Cent Hosp, Dept Resp Med, Gävle, Sweden.
    Brandén, Eva
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Centrum för klinisk forskning, Gävleborg. Gävle Cent Hosp, Dept Resp Med, Gävle, Sweden.
    Jirström, Karin
    Lund Univ, Dept Clin Sci, Div Oncol & Pathol, Lund, Sweden.
    Pontén, Fredrik
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Rahnenführer, Jörg
    TU Dortmund Univ, Dept Stat, Dortmund, Germany.
    Micke, Patrick
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Klinisk och experimentell patologi.
    Hengstler, Jan G
    TU Dortmund Univ, Leibniz Res Ctr Working Environm & Human Factors, Dortmund, Germany.
    Prognostic Impact of Tumor Cell Programmed Death Ligand 1 Expression and Immune Cell Infiltration in NSCLC2019Ingår i: Journal of Thoracic Oncology, ISSN 1556-0864, E-ISSN 1556-1380, Vol. 14, nr 4, s. 628-640, artikel-id S1556-0864(19)30009-7Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Introduction: Infiltration of T and B/plasma cells has been linked to NSCLC prognosis, but this has not been thoroughly investigated in relation to the expression of programmed death ligand 1 (PD-L1). Here, we determine the association of lymphocytes and PD-L1 with overall survival (OS) in two retrospective cohorts of operated NSCLC patients who were not treated with checkpoint inhibitors targeting the programmed death 1/PD-L1 axis. Moreover, we evaluate how PD-L1 positivity and clinicopathologic factors affect the prognostic association of lymphocytes.

    Methods: Cluster of differentiation (CD) 3 (CD3)-, CD8-, CD4-, forkhead box P3 (FOXP3)-, CD20-, CD79A-, and immunoglobulin kappa constant (IGKC)-positive immune cells, and tumor PD-L1 positivity, were determined by immunohistochemistry on tissue microarrays (n = 705). Affymetrix data was analyzed for a patient subset, and supplemented with publicly available transcriptomics data (N = 1724). Associations with OS were assessed by Kaplan-Meier plots and uni- and multivariate Cox regression.

    Results: Higher levels of T and B plasma cells were associated with longer OS (p = 0.004 and p < 0.001, for CD8 and IGKC, respectively). Highly proliferative tumors with few lymphocytes had the worst outcome. No association of PD-L1 positivity with OS was observed in a nonstratified patient population; however, a significant association with shorter OS was observed in never-smokers (p = 0.009 and p = 0.002, 5% and 50% cutoff). Lymphocyte infiltration was not associated with OS in PD-L1–positive tumors (50% cutoff). The prognostic association of lymphocyte infiltration also depended on the patients’ smoking history and histologic subtype.

    Conclusions: Proliferation, PD-L1 status, smoking history, and histology should be considered if lymphocyte infiltration is to be used as a prognostic biomarker.

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  • 334.
    Edwards, Steven J.
    et al.
    KTH Royal Inst Technol, Dept Appl Phys, Sci Life Lab, Stockholm, Sweden..
    Carannante, Valentina
    Karolinska Inst, Sci Life Lab, Dept Microbiol Tumor & Cell Biol, Stockholm, Sweden..
    Kuhnigk, Kyra
    Karolinska Inst, Dept Med Huddinge, Ctr Hematol & Regenerat Med, Stockholm, Sweden..
    Ring, Henrik
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för neurovetenskap, Hallböök: Stamceller, retinal utveckling och regeneration.
    Tararuk, Tatsiana
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för neurovetenskap, Hallböök: Stamceller, retinal utveckling och regeneration.
    Hallböök, Finn
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för neurovetenskap, Hallböök: Stamceller, retinal utveckling och regeneration.
    Blom, Hans
    KTH Royal Inst Technol, Dept Appl Phys, Sci Life Lab, Stockholm, Sweden..
    Önfelt, Björn
    KTH Royal Inst Technol, Dept Appl Phys, Sci Life Lab, Stockholm, Sweden..
    Brismar, Hjalmar
    KTH Royal Inst Technol, Dept Appl Phys, Sci Life Lab, Stockholm, Sweden..
    High-Resolution Imaging of Tumor Spheroids and Organoids Enabled by Expansion Microscopy2020Ingår i: Frontiers in Molecular Biosciences, E-ISSN 2296-889X, Vol. 7, artikel-id 208Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Three-dimensional cell cultures are able to better mimic the physiology and cellular environments found in tissuesin vivocompared to cells grown in two dimensions. In order to study the structure and function of cells in 3-D cultures, light microscopy is frequently used. The preparation of 3-D cell cultures for light microscopy is often destructive, including physical sectioning of the samples, which can result in the loss of 3-D information. In order to probe the structure of 3-D cell cultures at high resolution, we have explored the use of expansion microscopy and compared it to a simple immersion clearing protocol. We provide a practical method for the study of spheroids, organoids and tumor-infiltrating immune cells at high resolution without the loss of spatial organization. Expanded samples are highly transparent, enabling high-resolution imaging over extended volumes by significantly reducing light scatter and absorption. In addition, the hydrogel-like nature of expanded samples enables homogenous antibody labeling of dense epitopes throughout the sample volume. The improved labeling and image quality achieved in expanded samples revealed details in the center of the organoid which were previously only observable following serial sectioning. In comparison to chemically cleared spheroids, the improved signal-to-background ratio of expanded samples greatly improved subsequent methods for image segmentation and analysis.

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    FULLTEXT01
  • 335.
    Egaña, Isabel
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Kaito, Hiroshi
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Nitzsche, Anja
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Becker, Lore
    German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.; Department of Neurology, Friedrich-Baur-Institut, Ludwig-Maximilians-Universität München, Munich, Germany.
    Ballester-Lopez, Carolina
    German Res Ctr Environm Hlth GmbH, Helmholtz Zentrum Munchen, Inst Expt Genet, German Mouse Clin, Neuherberg, Germany.; German Ctr Lung Res DZL, Comprehens Pneumol Ctr, Helmholtz Zentrum Munchen, Inst Lung Biol & Dis,German Res Ctr Environm Hlth, Munich, Germany.
    Niaudet, Colin
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Petkova, Milena
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Liu, Wei
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper.
    Vanlandewijck, Michael
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Vernaleken, Alexandra
    German Res Ctr Environm Hlth GmbH, Helmholtz Zentrum Munchen, Inst Expt Genet, German Mouse Clin, Neuherberg, Germany.; Ludwig Maximilians Univ Munchen, Friedrich Baur Inst, Dept Neurol, Munich, Germany.
    Klopstock, Thomas
    Department of Neurology, Friedrich-Baur-Institut, Ludwig-Maximilians-Universität München, Munich, Germany.; German Center for Vertigo and Balance Disorders, Munich, Germany.; Deutsches Zentrum für Neurodegenerative Erkrankungen e. V. (DZNE), Munich, Germany.; German Network for Mitochondrial Disorders (mitoNET), Munich, Germany.
    Fuchs, Helmut
    German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
    Gailus-Durner, Valerie
    German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
    Hrabe de Angelis, Martin
    German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.; Tech Univ Munich, Ctr Life & Food Sci Weihenstephan, Expt Genet, Neuherberg, Germany.; German Ctr Diabet Res DZD, Neuherberg, Germany.
    Rask-Andersen, Helge
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper.
    Johansson, Henrik J.
    Karolinska Inst, Dept Oncol Pathol, Sci Life Lab, Canc Prote Mass Spectrometry, Stockholm, Sweden.
    Lehtiö, Janne
    Karolinska Inst, Dept Oncol Pathol, Sci Life Lab, Canc Prote Mass Spectrometry, Stockholm, Sweden.
    He, Liqun
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Yildirim, Ali Ö.
    German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.; Comprehensive Pneumology Center, Institute of Lung Biology and Disease, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), The German Center for Lung Research (DZL), Munich, Germany.
    Hellström, Mats
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi.
    Female mice lacking Pald1 exhibit endothelial cell apoptosis and emphysema2017Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 7, nr 15453Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Paladin (Pald1, mKIAA1274 or x99384) was identified in screens for vascular-specific genes and is a putative phosphatase. Paladin has also been proposed to be involved in various biological processes such as insulin signaling, innate immunity and neural crest migration. To determine the role of paladin we have now characterized the Pald1 knock-out mouse in a broad array of behavioral, physiological and biochemical tests. Here, we show that female, but not male, Pald1 heterozygous and homozygous knock-out mice display an emphysema-like histology with increased alveolar air spaces and impaired lung function with an obstructive phenotype. In contrast to many other tissues where Pald1 is restricted to the vascular compartment, Pald1 is expressed in both the epithelial and mesenchymal compartments of the postnatal lung. However, in Pald1 knock-out females, there is a specific increase in apoptosis and proliferation of endothelial cells, but not in non-endothelial cells. This results in a transient reduction of endothelial cells in the maturing lung. Our data suggests that Pald1 is required during lung vascular development and for normal function of the developing and adult lung in a sex-specific manner. To our knowledge, this is the first report of a sex-specific effect on endothelial cell apoptosis.

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  • 336.
    Eger, Nicole
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Advanced Detection Methods of Genomic Barcodes for Genotyping Escherichia coli Libraries2021Självständigt arbete på avancerad nivå (masterexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
    Abstract [en]

    Pooled cell strain libraries are a powerful tool allowing to investigate the influence of genetic modifications on phenotypes in high throughput single-cell assays. To link the genotype to phenotype in each cell of the library, unique 20 base pairs (bp) long barcodes are used to allow in situ genotyping after phenotyping via fluorescence microscopy. In previous studies, these barcode sequences were expressed from high copy number plasmids resulting in a high number of targets for detection via fluorescence in situ hybridization (FISH) and thus, a strong readout signal. However, constant selection pressure must be applied on the cells to maintain the foreign plasmid DNA which may influence the phenotype. Inserting unique barcodes on the chromosome ensures stability of the construct which is required for some genomic library applications. However, the low copy number of the barcode sequence often requires an additional step of DNA amplification for efficient detection. In this study, two methods for barcode amplification were investigated. First, amplification from the double stranded DNA upon binding of peptide nucleic acids and subsequent amplification via rolling circle amplification (AmPPR). Second, amplification from genomic DNA or cDNA via loop-mediated isothermal amplification (LAMP). Whereas the AmPPR approach remained unsuccessful, chromosomal barcode sequences were successfully amplified in situ via LAMP and subsequently detected using FISH. I show that LAMP can potentially be a quick, specific, and elegant amplification technique for in situ genotyping in microfluidic devices. However, nonspecific amplification and partly nonspecific readout signals when using LAMP remain a problem and need to be further investigated before implementing this method on pooled libraries.

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  • 337.
    Egevad, Lars
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Frimmel, Hans
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
    Mattson, Stefan
    Uppsala universitet, Humanistisk-samhällsvetenskapliga vetenskapsområdet, Samhällsvetenskapliga fakulteten, Institutionen för informationsvetenskap, Statistik.
    Bengtsson, Ewert
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
    Busch, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Biopsy protocol stability in a three-dimensional model of prostate cancer: Changes in cancer yield after adjustment of biopsy positions1999Ingår i: Urology, ISSN 0090-4295, E-ISSN 1527-9995, Vol. 54, s. 862-868Artikel i tidskrift (Refereegranskat)
  • 338.
    Egevad, Lars
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Frimmel, Hans
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
    Norberg, Mona
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för onkologi, radiologi och klinisk immunologi, Enheten för radiologi.
    Mattson, Stefan
    Uppsala universitet, Humanistisk-samhällsvetenskapliga vetenskapsområdet, Samhällsvetenskapliga fakulteten, Institutionen för informationsvetenskap, Statistik.
    Carlbom, Ingrid
    Bengtsson, Ewert
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
    Busch, Christer
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Three-dimensional computer reconstruction of prostate cancer from radical prostatectomy specimens: Evaluation of the model by core biopsy simulation1999Ingår i: Urology, ISSN 0090-4295, E-ISSN 1527-9995, Vol. 53, s. 192-198Artikel i tidskrift (Refereegranskat)
  • 339.
    Einarsson, Elin
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Mikrobiologi.
    Ma'ayeh, Showgy Y.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Mikrobiologi.
    Svärd, Staffan G.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Mikrobiologi.
    An up-date on Giardia and giardiasis2016Ingår i: Current Opinion in Microbiology, ISSN 1369-5274, E-ISSN 1879-0364, Vol. 34, s. 47-52Artikel, forskningsöversikt (Refereegranskat)
    Abstract [en]

    Giardia intestinalis is a non-invasive protozoan parasite infecting the upper small intestine causing acute, watery diarrhea or giardiasis in 280 million people annually. Asymptomatic infections are equally common and recent data have suggested that infections even can be protective against other diarrhea! diseases. Most symptomatic infections resolve spontaneously but infections can lead to chronic disease and treatment failures are becoming more common world-wide. Giardia infections can also result in irritable bowel syndrome (IBS) and food allergies after resolution. Until recently not much was known about the mechanism of giardiasis or the cause of post-giardiasis syndromes and treatment failures, but here we will describe the recent progress in these areas.

  • 340.
    Ejdesjö, Andreas
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Genetic predisposition for malformations in diabetic rat pregnancy2008Licentiatavhandling, sammanläggning (Övrigt vetenskapligt)
    Delarbeten
    1. Genetic and environmental influence on diabetic rat embryopathy
    Öppna denna publikation i ny flik eller fönster >>Genetic and environmental influence on diabetic rat embryopathy
    2011 (Engelska)Ingår i: American Journal of Physiology. Endocrinology and Metabolism, ISSN 0193-1849, E-ISSN 1522-1555, Vol. 300, nr 3, s. E454-E467Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    We assessed genetic and environmental influence on fetal outcome in diabetic rat pregnancy. Crossing normal (N) and manifestly diabetic (MD) Wistar Furth (W) and Sprague-Dawley (L) females with W or L males yielded 4 different fetal genotypes (WW, LL, WL, LW) in N or MD rat pregnancies for studies. We also evaluated fetal outcome in litters with enhanced or diminished severity of maternal MD state, denoted MD(+)WL and MD(-)LW. The MDWW litters had less malformations and resorptions (0% and 19%) than the MDLL litters (17% and 30%). The MDWL litters (0% and 8%) were less maldeveloped than the MDLW litters (9% and 22%), whereas the MD(+)WL (3% and 23%) and MD(-)LW (1% and 17%) litters showed increased and decreased dysmorphogenesis (compared to MDWL and MDLW litters). The pregnant MDW rats had lower serum levels of glucose, fructosamine and branched chain amino acids than the pregnant MDL rats, whereas the pregnant MD(+)W and MD(-)L rats had levels comparable to those of the MDL and MDW rats, respectively. The 8-iso-PGF2α levels of the malformed MDLW offspring were increased compared to the non-malformed MDLW offspring. Diabetes decreased fetal heart Ret and increased Bmp-4 gene expression in the MDLW offspring, and caused decreased GDNF and Shh expression in the malformed fetal mandible of the MDLW offspring. We conclude that the fetal (epi)genome controls the embryonic dysmorphogenesis in diabetic pregnancy by instigating a threshold level for the teratological insult, and that the maternal genome controls the teratogenic insult by (dys)regulating the maternal metabolism.

    Nyckelord
    diabetes in pregnancy, congenital abnormalities, teratology, animal experimentation, aldose reductase, glyceraldehyde-3-phosphate dehydrogenase, sonic hedgehog homologue, ret proto-oncogene, glial-derived neurotrophic factor, antioxidative enzymes
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-105013 (URN)10.​1152/​ajpendo.​00543.​2010 (DOI)000287796200004 ()21119026 (PubMedID)
    Tillgänglig från: 2009-05-31 Skapad: 2009-05-31 Senast uppdaterad: 2022-01-28Bibliografiskt granskad
    2. Epigenetic impact on offspring in diabetic rat pregnancy
    Öppna denna publikation i ny flik eller fönster >>Epigenetic impact on offspring in diabetic rat pregnancy
    (Engelska)Manuskript (Övrigt vetenskapligt)
    Identifikatorer
    urn:nbn:se:uu:diva-105016 (URN)
    Tillgänglig från: 2009-05-31 Skapad: 2009-05-31 Senast uppdaterad: 2011-11-02
  • 341.
    Ejdesjö, Andreas
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Teratogenic Predisposition in Diabetic Rat Pregnancy2012Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Pre-gestational diabetes increases the risk of congenital malformation in the offspring and both morbidity and mortality in the diabetic mother and her offspring. During pregnancy, high glucose levels act as a teratogen through several cellular and biochemical pathways and increased production of reactive oxygen species (ROS) has a central role in diabetic embryopathy. The aim of this work was to investigate the importance of genetic predisposition for congenital malformations and to study the genes involved in the teratogenic process of diabetic pregnancy.

    The crossbreeding of two rat strains, with both low and high incidence of diabetes-induced malformations, indicated that strain-specific maternal factors, such as disturbed serum levels of amino acids, triglycerides, and β-hydroxybutyrate, were associated with malformation. In addition, disturbed fetal expression of genes involved in ROS defense and development (Shh, Bmp4, Ret and Gdnf) in mandible and heart, and decreased activity of Gapdh and Aldose Reductase were associated with the teratogenic process, and the trans-generational heredity of the mother determined the type of malformations induced by maternal diabetes.

    In rat embryos, a diabetic environment in utero changed the expression of genes involved in ROS defense (Nrf2, Gpx1 and Cat), development of mandible and heart (Msx2, Shh, Bmp4, Ret and Gdnf), and neural tube closure and apoptosis (Pax3 and p53). The changes were divergent with tissue-specific alterations of gene expression in developing mandible, heart anlage, and whole embryo.

    Disruption of the Receptor for Advanced Glycation End products (RAGE) had a protective effect against diabetic embryopathy in mice, and the blockage of RAGE diminished ROS production in the offspring: this supported oxidative stress being a necessary etiological component in diabetic embryopathy.

    Maternal metabolic state and genetic susceptibility influence fetal outcome in experimental diabetic pregnancy. Disturbed protection against oxidative stress and tissue-specific derangements in the expression of developmental genes play pivotal roles in the teratogenic mechanism, and enhanced levels of Advanced Glycation End products (AGE) and RAGE-induced oxidative stress are involved in diabetic dysmorphogenesis.

    Delarbeten
    1. Genetic and environmental influence on diabetic rat embryopathy
    Öppna denna publikation i ny flik eller fönster >>Genetic and environmental influence on diabetic rat embryopathy
    2011 (Engelska)Ingår i: American Journal of Physiology. Endocrinology and Metabolism, ISSN 0193-1849, E-ISSN 1522-1555, Vol. 300, nr 3, s. E454-E467Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    We assessed genetic and environmental influence on fetal outcome in diabetic rat pregnancy. Crossing normal (N) and manifestly diabetic (MD) Wistar Furth (W) and Sprague-Dawley (L) females with W or L males yielded 4 different fetal genotypes (WW, LL, WL, LW) in N or MD rat pregnancies for studies. We also evaluated fetal outcome in litters with enhanced or diminished severity of maternal MD state, denoted MD(+)WL and MD(-)LW. The MDWW litters had less malformations and resorptions (0% and 19%) than the MDLL litters (17% and 30%). The MDWL litters (0% and 8%) were less maldeveloped than the MDLW litters (9% and 22%), whereas the MD(+)WL (3% and 23%) and MD(-)LW (1% and 17%) litters showed increased and decreased dysmorphogenesis (compared to MDWL and MDLW litters). The pregnant MDW rats had lower serum levels of glucose, fructosamine and branched chain amino acids than the pregnant MDL rats, whereas the pregnant MD(+)W and MD(-)L rats had levels comparable to those of the MDL and MDW rats, respectively. The 8-iso-PGF2α levels of the malformed MDLW offspring were increased compared to the non-malformed MDLW offspring. Diabetes decreased fetal heart Ret and increased Bmp-4 gene expression in the MDLW offspring, and caused decreased GDNF and Shh expression in the malformed fetal mandible of the MDLW offspring. We conclude that the fetal (epi)genome controls the embryonic dysmorphogenesis in diabetic pregnancy by instigating a threshold level for the teratological insult, and that the maternal genome controls the teratogenic insult by (dys)regulating the maternal metabolism.

    Nyckelord
    diabetes in pregnancy, congenital abnormalities, teratology, animal experimentation, aldose reductase, glyceraldehyde-3-phosphate dehydrogenase, sonic hedgehog homologue, ret proto-oncogene, glial-derived neurotrophic factor, antioxidative enzymes
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-105013 (URN)10.​1152/​ajpendo.​00543.​2010 (DOI)000287796200004 ()21119026 (PubMedID)
    Tillgänglig från: 2009-05-31 Skapad: 2009-05-31 Senast uppdaterad: 2022-01-28Bibliografiskt granskad
    2. Influence of maternal metabolism and parental genetics on fetal maldevelopment in diabetic rat pregnancy
    Öppna denna publikation i ny flik eller fönster >>Influence of maternal metabolism and parental genetics on fetal maldevelopment in diabetic rat pregnancy
    2012 (Engelska)Ingår i: American Journal of Physiology. Endocrinology and Metabolism, ISSN 0193-1849, E-ISSN 1522-1555, Vol. 302, nr 10, s. E1198-E1209Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The purpose of this study was to investigate the influence of parental transgenerational genetics and maternal metabolic state on fetal maldevelopment in diabetic rat pregnancy. Rats from an inbred malformation-resistant (W) strain, and an inbred malformation-prone (L) strain, were cross-mated to produce two different F-1 hybrids, WL and LW. Normal (N) and manifestly diabetic (MD) WL and LW females were mated with normal males of the same F1 generation to obtain WLWL and LWLW F-2 hybrids. Maternal diabetes increased malformation and resorption rates in both F-2 generations. MD-WLWL offspring had higher resorption rate but similar malformation rate compared with the MD-LWLW offspring. Malformed MD-WLWL offspring presented with 100% agnathia/micrognathia, whereas malformed MD-LWL offspring had 60% agnathia/micrognathia and 40% cleft lip and palate. The MD-WL dams showed increased beta-hydroxy-butyrate levels and alterations in concentrations of several amino acids (taurine, asparagine, citrulline, cystine, glutamic acid, leucine, tyrosine, and tryptophan) compared with MD-LW dams. Fetal glyceraldehyde-3-phosphate dehydrogenase (Gapdh) activity and gene expression were more altered in MD-WLWL than MD-LWLW. Fetal gene expression of reactive oxygen species (ROS) scavenger enzymes was diminished in MD-WLWL compared with MD-LWLW. Glial cell line-derived neurotrophic factor and Ret proto-oncogene gene expression was decreased in both MD-WLWL and MD-LWLW fetuses, whereas increased bone morphogenetic protein 4 and decreased Sonic hedgehog homolog expression was found only in MD-LWLW fetuses. Despite identical autosomal genotypes, the WL and LW dams gave birth to offspring with markedly different malformation patterns. Together with fetal differences in enzymatic activity and expression of Gapdh, ROS scavengers, and developmental genes, these results may suggest a teratological mechanism in diabetic pregnancy influenced by maternal metabolism and parental strain epigenetics.

    Nyckelord
    diabetes, malformations, teratogenic, gene expression
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-176242 (URN)10.1152/ajpendo.00661.2011 (DOI)000304360400005 ()
    Tillgänglig från: 2012-06-19 Skapad: 2012-06-18 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    3. Alterations in the expression of tissue specific genes of the developing mandible and heart in rat diabetic embryopathy
    Öppna denna publikation i ny flik eller fönster >>Alterations in the expression of tissue specific genes of the developing mandible and heart in rat diabetic embryopathy
    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Abstract [en]

    Background & Aim: Maternal diabetes induces skeletal and cardiac malformations in the offspring, both in human and experimental diabetic pregnancies. A developmental disturbance in neural crest cell (NCC) derived embryonic tissue is a plausible origin for these malformations. Thus, diabetes-exposed and control rat embryos from a locally outbred Sprague-Dawley rat strain were used to investigate tissue-specific alterations in the expression of suggested candidate genes in the developing mandible and heart anlage and in the whole embryo.

    Methods: Female non-diabetic (N) and streptozotocin-induced manifestly diabetic (MD) rats were mated with N males. Embryos were collected and morphologically examined on gestational days (GD) 11 and 13. The developing mandible (first pharyngeal arch), heart anlage and the remaining embryonic tissues were prepared and analyzed with quantitative real time PCR.

    Results: Maternal diabetes changed the gene expression in the developing mandible where Gpx1 (MD11) and Cat (MD13) decreased. In the heart anlage, diabetes decreased Nrf2 (MD11), whereas, in whole embryo, diabetes increased Nrf2 (MD13). Maternal diabetes changed the gene expression in the developing mandible, Bmp4 (MD13) decreased, and Gdnf (MD11 and MD13) and Ret (MD11 and MD13) increased. In the heart anlage, diabetes decreased Shh (MD11) and Gdnf (MD11 and MD13). In whole embryo, diabetes decreased Shh (MD11) and Gdnf (MD13) and increased Msx2 (MD13) and Ret (MD11 and MD13). Maternal diabetes changed the gene expression in the developing mandible; Pax3 was both increased (MD11) and decreased (MD13). In the heart anlage, diabetes decreased Pax3 (MD13), whereas, in the whole embryo, diabetes increased Pax3 (MD13) and both decreased p53 (MD11) and increased p53 (MD13).

    Conclusions: Hyperglycemia in utero causes tissue-specific alterations in embryonic gene expression of several important antioxidative defense and developmental genes. Tissuespecific disturbance of gene expressions suggests a diminished ROS scavenging capacity and a role for altered gene expression of Gdnf, Ret, Bmp4 and Pax3 in the diabetes-induced embryonic dysmorphogenesis.

    Nyckelord
    diabetes, malformations, teratogenic, gene expression
    Nationell ämneskategori
    Cell- och molekylärbiologi
    Forskningsämne
    Medicinsk cellbiologi
    Identifikatorer
    urn:nbn:se:uu:diva-178168 (URN)
    Tillgänglig från: 2012-07-30 Skapad: 2012-07-30 Senast uppdaterad: 2018-01-12
    4. Receptor for Advanced Glycation End products (RAGE) knockout reduces fetal dysmorphogenesis in murine diabetic pregnancy
    Öppna denna publikation i ny flik eller fönster >>Receptor for Advanced Glycation End products (RAGE) knockout reduces fetal dysmorphogenesis in murine diabetic pregnancy
    Visa övriga...
    2016 (Engelska)Ingår i: Reproductive Toxicology, ISSN 0890-6238, E-ISSN 1873-1708, Vol. 62, s. 62-70Artikel i tidskrift (Övrigt vetenskapligt) Published
    Abstract [en]

    Background & Aim: The receptor for Advanced Glycation End products (RAGE) is implicated in the pathogenesis of diabetic complications, but its importance for the induction of congenital malformations in diabetic pregnancy is unclear. The aim of the present study was to investigate a possible role of RAGE activation in the induction of diabetic embryopathy.

    Methods: Female non-diabetic and diabetic wildtype (WT) C57Bl/6 mice and RAGE knockout C57Bl/6 (RAGE‑/-) mice were mated with males of the same genotype. Diabetes was induced by daily streptozotocin (STZ) injections (50 mg/kg STZ i.p.) on five consecutive days. On gestational day 18, pregnant mice were anesthetized and blood was drawn from the heart to measure maternal metabolic parameters. Fetuses and placentas were excised, weighed, and examined for morphological anomalies, and fetal livers were analyzed for 8‑iso‑PGF levels.

    Results: There were no malformations in non-diabetic WT or non-diabetic RAGE‑/- mice. However, resorption rates were higher in non-diabetic WT (10%) than in non-diabetic RAGE‑/- mice (4%). Diabetic WT mice had higher malformation (22%) and resorption (43%) rates than diabetic RAGE‑/- mice (3% malformations and 21% resorptions). Maternal diabetes decreased fetal weight more in WT fetuses (44%) than in RAGE‑/- fetuses (36%). There were no differences in plasma glucose levels between the diabetic WT and RAGE‑/- mice, but plasma levels of triglycerides and cholesterol were lower in diabetic WT mice than in diabetic RAGE-/- mice. Diabetes increased maternal plasma levels of methylglyoxal in WT and RAGE‑/- mice, and increased fetal hepatic levels of 8-iso-PGF in WT fetuses, but not in RAGE‑/- fetuses.

    Discussion: Knockout of RAGE diminished the rates of fetal malformations and resorptions, despite similar levels of hyperglycemia in pregnant diabetic mice. An anti-teratogenic effect was present in RAGE‑/- mice despite having a more severe diabetic state than diabetic WT mice. As 8-iso-PGF, a marker of oxidative stress, only increased in diabetic WT offspring, this suggested a pivotal role of RAGE activation and oxidative stress in the pathogenesis of diabetic embryopathy.

    Nyckelord
    Diabetes; AGE; RAGE; Oxidative stress; Diabetic embryopathy; Neural tube defects; Experimental teratology
    Nationell ämneskategori
    Cell- och molekylärbiologi
    Forskningsämne
    Medicinsk cellbiologi
    Identifikatorer
    urn:nbn:se:uu:diva-178170 (URN)10.1016/j.reprotox.2016.04.015 (DOI)000378367700008 ()27109771 (PubMedID)
    Forskningsfinansiär
    DiabetesförbundetNovo NordiskVetenskapsrådet, 54X-21117Deutsche Forschungsgemeinschaft (DFG), SFB1118
    Tillgänglig från: 2012-07-30 Skapad: 2012-07-30 Senast uppdaterad: 2018-01-12Bibliografiskt granskad
    Ladda ner fulltext (pdf)
    fulltext
  • 342.
    Ejdesjö, Andreas
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Brings, Sebastian
    Fleming, Thomas
    Fred, Rikard G.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Nawroth, Peter P
    University of Heidelberg.
    Eriksson, Ulf J
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Receptor for Advanced Glycation End products (RAGE) knockout reduces fetal dysmorphogenesis in murine diabetic pregnancy2016Ingår i: Reproductive Toxicology, ISSN 0890-6238, E-ISSN 1873-1708, Vol. 62, s. 62-70Artikel i tidskrift (Övrigt vetenskapligt)
    Abstract [en]

    Background & Aim: The receptor for Advanced Glycation End products (RAGE) is implicated in the pathogenesis of diabetic complications, but its importance for the induction of congenital malformations in diabetic pregnancy is unclear. The aim of the present study was to investigate a possible role of RAGE activation in the induction of diabetic embryopathy.

    Methods: Female non-diabetic and diabetic wildtype (WT) C57Bl/6 mice and RAGE knockout C57Bl/6 (RAGE‑/-) mice were mated with males of the same genotype. Diabetes was induced by daily streptozotocin (STZ) injections (50 mg/kg STZ i.p.) on five consecutive days. On gestational day 18, pregnant mice were anesthetized and blood was drawn from the heart to measure maternal metabolic parameters. Fetuses and placentas were excised, weighed, and examined for morphological anomalies, and fetal livers were analyzed for 8‑iso‑PGF levels.

    Results: There were no malformations in non-diabetic WT or non-diabetic RAGE‑/- mice. However, resorption rates were higher in non-diabetic WT (10%) than in non-diabetic RAGE‑/- mice (4%). Diabetic WT mice had higher malformation (22%) and resorption (43%) rates than diabetic RAGE‑/- mice (3% malformations and 21% resorptions). Maternal diabetes decreased fetal weight more in WT fetuses (44%) than in RAGE‑/- fetuses (36%). There were no differences in plasma glucose levels between the diabetic WT and RAGE‑/- mice, but plasma levels of triglycerides and cholesterol were lower in diabetic WT mice than in diabetic RAGE-/- mice. Diabetes increased maternal plasma levels of methylglyoxal in WT and RAGE‑/- mice, and increased fetal hepatic levels of 8-iso-PGF in WT fetuses, but not in RAGE‑/- fetuses.

    Discussion: Knockout of RAGE diminished the rates of fetal malformations and resorptions, despite similar levels of hyperglycemia in pregnant diabetic mice. An anti-teratogenic effect was present in RAGE‑/- mice despite having a more severe diabetic state than diabetic WT mice. As 8-iso-PGF, a marker of oxidative stress, only increased in diabetic WT offspring, this suggested a pivotal role of RAGE activation and oxidative stress in the pathogenesis of diabetic embryopathy.

  • 343.
    Ejdesjö, Andreas
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Wentzel, Parri
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Eriksson, Ulf J
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Alterations in the expression of tissue specific genes of the developing mandible and heart in rat diabetic embryopathyManuskript (preprint) (Övrigt vetenskapligt)
    Abstract [en]

    Background & Aim: Maternal diabetes induces skeletal and cardiac malformations in the offspring, both in human and experimental diabetic pregnancies. A developmental disturbance in neural crest cell (NCC) derived embryonic tissue is a plausible origin for these malformations. Thus, diabetes-exposed and control rat embryos from a locally outbred Sprague-Dawley rat strain were used to investigate tissue-specific alterations in the expression of suggested candidate genes in the developing mandible and heart anlage and in the whole embryo.

    Methods: Female non-diabetic (N) and streptozotocin-induced manifestly diabetic (MD) rats were mated with N males. Embryos were collected and morphologically examined on gestational days (GD) 11 and 13. The developing mandible (first pharyngeal arch), heart anlage and the remaining embryonic tissues were prepared and analyzed with quantitative real time PCR.

    Results: Maternal diabetes changed the gene expression in the developing mandible where Gpx1 (MD11) and Cat (MD13) decreased. In the heart anlage, diabetes decreased Nrf2 (MD11), whereas, in whole embryo, diabetes increased Nrf2 (MD13). Maternal diabetes changed the gene expression in the developing mandible, Bmp4 (MD13) decreased, and Gdnf (MD11 and MD13) and Ret (MD11 and MD13) increased. In the heart anlage, diabetes decreased Shh (MD11) and Gdnf (MD11 and MD13). In whole embryo, diabetes decreased Shh (MD11) and Gdnf (MD13) and increased Msx2 (MD13) and Ret (MD11 and MD13). Maternal diabetes changed the gene expression in the developing mandible; Pax3 was both increased (MD11) and decreased (MD13). In the heart anlage, diabetes decreased Pax3 (MD13), whereas, in the whole embryo, diabetes increased Pax3 (MD13) and both decreased p53 (MD11) and increased p53 (MD13).

    Conclusions: Hyperglycemia in utero causes tissue-specific alterations in embryonic gene expression of several important antioxidative defense and developmental genes. Tissuespecific disturbance of gene expressions suggests a diminished ROS scavenging capacity and a role for altered gene expression of Gdnf, Ret, Bmp4 and Pax3 in the diabetes-induced embryonic dysmorphogenesis.

  • 344.
    Ekman, Maria
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwiginstitutet för cancerforskning.
    The role of Smad7 and TRAF6 in Prostate Cancer Cell Invasion, Migration and Survival2011Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Transforming growth factor (TGF) β is a tumor suppressor during early tumor development, by inhibiting proliferation and inducing apoptosis. At later stages of cancer, it becomes a tumor promoter, and promotes tumor cell migration and invasion. TGFβ signals via its type II and type I receptors to several downstream signaling pathways. In the present work we have focused on the TRAF6 (tumor necrosis factor receptor-associated factor 6)/ TAK1 (TGFβ activated kinase 1) signaling pathway and the Smad7-dependent activation of p38 in prostate carcinoma cells (PC3U). We found that TGFβ-induced activation of the ubiquitin ligase TRAF6 was needed for cell invasion, by a mechanism that involves activation of the metalloproteinase TNFα converting enzyme (TACE), via protein kinase Cζ (PKCζ). TACE cleaves the TβRI, whereafter the intracellular domain (ICD) translocates to the nucleus, where it binds to the transcriptional co-activator p300 and regulates gene expression, promoting invasion. Interestingly, the translocation of the TβRI ICD was observed in several cancer cell lines and in sections of primary tumors, but not in primary prostate epithelial cells. We also found that Smad7 and adenomatous polyposis coli (APC) are important for TGFβ- and epidermal growth factor (EGF)-induced cell migration in PC3U cells. TGFβ induces the formation of a complex consisting of Smad7, p38, glycogene synthase kinase 3β (GSK-3β), APC and β-catenin, which localizes to the membrane ruffles in the leading edge of migrating cells. The complex links the TβRI to the microtubule system and promotes membrane ruffling and microtubule polarization, which are known to be important for cell migration. In the EGF signaling pathway, Smad7 was found to be important for phosphorylation of the EGF receptor at Tyr1068, for the activation of p38 and JNK, and for induction of membrane ruffles. Smad7 is required for TGFβ-induced activation of p38 and apoptosis. We found that Smad7 forms a complex with p38 and ataxia telangiectasia mutated (ATM), which is important for activation of p53 mediated apoptosis. Many tumor cells including the PC3U cells lack a functional p53, which is one of the reasons to why cancer cells can avoid the tumor suppressor effects of TGFβ.

    Delarbeten
    1. TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer
    Öppna denna publikation i ny flik eller fönster >>TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer
    Visa övriga...
    2011 (Engelska)Ingår i: Nature Communications, E-ISSN 2041-1723, Vol. 2, nr 330Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Transforming growth factor β (TGFβ) is a pluripotent cytokine promoting epithelial cell plasticity during morphogenesis and tumour progression. TGFβ binding to type II and type I serine/threonine kinase receptors (TβRII and TβRI) causes activation of different intracellular signaling pathways. TβRI is associated with the ubiquitin ligase tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6). Here we show that TGFβ, via TRAF6, causes Lys63-linked polyubiquitination of TβRI, promoting cleavage of TβRI by TNF-alpha converting enzyme (TACE), in a PKCζ-dependent manner. The liberated intracellular domain (ICD) of TβRI associates with the transcriptional regulator p300 to activate genes involved in tumour cell invasiveness, such as Snail and MMP2. Moreover, TGFβ-induced invasion of cancer cells is TACE- and PKCζ- dependent and the TβRI ICD is localized in the nuclei of different kinds of tumour cells in tissue sections. Thus, our data reveal a specific role for TβRI in TGFβ mediated tumour invasion.

    Ort, förlag, år, upplaga, sidor
    Macmillan Publishers Limited, 2011
    Nyckelord
    TRAF6, TGF-beta, TGF-beta receptor I, cleavage, TACE, atypical PKC
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Forskningsämne
    Cellforskning
    Identifikatorer
    urn:nbn:se:uu:diva-159148 (URN)10.1038/ncomms1332 (DOI)000294802600035 ()
    Tillgänglig från: 2011-09-22 Skapad: 2011-09-22 Senast uppdaterad: 2023-03-28Bibliografiskt granskad
    2. APC and Smad7 link the TGFβ type I receptors to the microtubule system to promote cell migration
    Öppna denna publikation i ny flik eller fönster >>APC and Smad7 link the TGFβ type I receptors to the microtubule system to promote cell migration
    Visa övriga...
    2012 (Engelska)Ingår i: Molecular Biology of the Cell, ISSN 1059-1524, E-ISSN 1939-4586, Vol. 23, nr 11, s. 2109-2121Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Cell migration occurs by activation of complex regulatory pathways that are spatially and temporally integrated in response to extracellular cues. Binding of adenomatous polyposis coli (APC) to the microtubule plus ends in polarized cells is regulated by glycogen synthase kinase 3 beta (GSK-3 beta). This event is crucial for establishment of cell polarity during directional migration. However, the role of APC for cellular extension in response to extracellular signals is less clear. Smad7 is a direct target gene for transforming growth factor-beta (TGF beta) and is known to inhibit various TGF beta-induced responses. Here we report a new function for Smad7. We show that Smad7 and p38 mitogen-activated protein kinase together regulate the expression of APC and cell migration in prostate cancer cells in response to TGF beta stimulation. In addition, Smad7 forms a complex with APC and acts as an adaptor protein for p38 and GSK-3 beta kinases to facilitate local TGF beta/p38-dependent inactivation of GSK-3 beta, accumulation of beta-catenin, and recruitment of APC to the microtubule plus end in the leading edge of migrating prostate cancer cells. Moreover, the Smad7-APC complex links the TGF beta type I receptor to the microtubule system to regulate directed cellular extension and migratory responses evoked by TGF beta.

    Nyckelord
    APC, Smad7, GSK-3beta, p38, TGF-beta, cell migration
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Forskningsämne
    Cellforskning
    Identifikatorer
    urn:nbn:se:uu:diva-159146 (URN)10.1091/mbc.E10-12-1000 (DOI)000306286400008 ()
    Tillgänglig från: 2011-09-22 Skapad: 2011-09-22 Senast uppdaterad: 2022-01-28Bibliografiskt granskad
    3. Smad7 and APC are required for EGF-induced cell migration in human prostate epithelial cells
    Öppna denna publikation i ny flik eller fönster >>Smad7 and APC are required for EGF-induced cell migration in human prostate epithelial cells
    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Nyckelord
    EGF, EGFR, Smad7, APC, cell migration
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Forskningsämne
    Cellforskning
    Identifikatorer
    urn:nbn:se:uu:diva-159149 (URN)
    Tillgänglig från: 2011-09-22 Skapad: 2011-09-22 Senast uppdaterad: 2011-11-04
    4. TGF beta 1-induced activation of ATM and p53 mediates apoptosis in a Smad7-dependent manner
    Öppna denna publikation i ny flik eller fönster >>TGF beta 1-induced activation of ATM and p53 mediates apoptosis in a Smad7-dependent manner
    Visa övriga...
    2006 (Engelska)Ingår i: Cell Cycle, ISSN 1538-4101, E-ISSN 1551-4005, Vol. 5, nr 23, s. 2787-2795Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    ATM, a DNA-damage sensitive kinase and p53, are frequently inactivated in a variety of cancers as they together with gamma H2AX are critical guardians against DNA damage. Here, we report of a functional cross-talk between the cytokine TGF beta and p53, leading to apoptosis of epithelial cells, involving Smad7, a TGF beta target gene, p38 MAP kinase, and ATM. Using ectopic expression of p53, siRNA for Smad7, p38a(-/-) deficient cells and specific inhibitors, we show that TGF-beta induces apoptosis via ATM and p53 in epithelial cells. Intriguingly, Smad7 act as a scaffold protein to promote functional interactions between p38, ATM and p53 upon TGF beta treatment, facilitating their activation. Smad7. colocalizes with gamma H2AX in DNA damage foci and was required for proper cell cycle checkpoints to prevent genetic instability. Our data imply that Smad7 plays a crucial role upstream of ATM and p53 to protect the genome from insults evoked by extracellular stress.

    Nyckelord
    apoptosis, prostate cancer, p53, Smad, ATM
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-147029 (URN)000242897800016 ()17172861 (PubMedID)
    Tillgänglig från: 2011-02-23 Skapad: 2011-02-23 Senast uppdaterad: 2017-12-11Bibliografiskt granskad
    Ladda ner fulltext (pdf)
    fulltext
  • 345.
    El-Aarag, Bishoy
    et al.
    Menoufia Univ, Fac Sci, Chem Dept, Biochem Div, Shibin Al Kawm 32512, Egypt;Okayama Univ, Grad Sch Nat Sci & Technol, Div Chem & Biotechnol, Okayama 7008530, Japan.
    Khairy, Asmaa
    Menoufia Univ, Fac Sci, Chem Dept, Shibin Al Kawm 32512, Egypt.
    Khalifa, Shaden A. M.
    Stockholm Univ, Wenner Gren Inst, Dept Mol Biosci, SE-10691 Stockholm, Sweden;Novum, Dept Expt Canc Med ECM, S-14157 Huddinge, Sweden.
    El-Seedi, Hesham
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Farmaceutiska fakulteten, Institutionen för läkemedelskemi, Farmakognosi. Menoufia Univ, Fac Sci, Chem Dept, Shibin Al Kawm 32512, Egypt;Jiangsu Univ, Int Res Ctr Food Nutr & Safety, Zhenjiang 212013, Jiangsu, Peoples R China;Al Rayan Coll, Al Rayan Res & Innovat Ctr, Medina 42541, Saudi Arabia.
    Protective Effects of Flavone from Tamarix aphylla against CCl4-Induced Liver Injury in Mice Mediated by Suppression of Oxidative Stress, Apoptosis and Angiogenesis2019Ingår i: International Journal of Molecular Sciences, ISSN 1661-6596, E-ISSN 1422-0067, Vol. 20, nr 20, artikel-id 5215Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The current study aimed to investigate, for the first time, the beneficial effects of 3,5-dihydroxy-4',7-dimethoxyflavone isolated from Tamarix aphylla L. against liver injury in mice. Liver injury was induced by intraperitoneal (i.p.) injection of carbon tetrachloride (CCl4) at a dose of 0.4 mL/kg mixed in olive oil at ratio (1:4) twice a week for 6 consecutive weeks. The administration of CCl4 caused significant histopathological changes in liver tissues while the pre-treatment with the flavone at dose of 10 and 25 mg/kg ameliorated the observed liver damages. Also, it markedly reduced hepatic malondialdehyde (MDA) level as well as increased the activities of liver superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (Gpx) compared with their recorded levels in CCl4 model group. Moreover, the immunohistochemical analysis demonstrated the enhancement in the protein level of B-cell lymphoma-2 (Bcl-2) while the protein levels of cysteine-aspartic acid protease-3 (caspase-3), Bcl-2-associated x protein (Bax), transforming growth factor-beta 1 (TGF-beta 1) and CD31 were suppressed following the flavone treatement. These results suggest that the flavone can inhibit liver injury induced in mice owning to its impact on the oxidation, apoptotic and angiogenesis mechanisms. Further pharmacological investigations are essential to determine the effectiveness of the flavone in human.

    Ladda ner fulltext (pdf)
    FULLTEXT01
  • 346.
    El-Aarag, Bishoy
    et al.
    Menoufia Univ, Fac Sci, Chem Dept, Biochem Div, Shibin Al Kawm 32512, Egypt;Okayama Univ, Grad Sch Nat Sci & Technol, Div Chem & Biotechnol, Okayama 7008530, Japan.
    Magdy, Mohamed
    Menoufia Univ, Dept Chem, Fac Sci, Shibin Al Kawm 32512, Egypt.
    AlAjmi, Mohamed F.
    King Saud Univ, Dept Pharmacognosy, Coll Pharm, Riyadh 11451, Saudi Arabia.
    Khalifa, Shaden A. M.
    Stockholm Univ, Wenner Gren Inst, Dept Mol Biosci, S-10691 Stockholm, Sweden.
    El-Seedi, Hesham
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Farmaceutiska fakulteten, Institutionen för läkemedelskemi, Farmakognosi. Menoufia Univ, Dept Chem, Fac Sci, Shibin Al Kawm 32512, Egypt;Univ Karachi, Int Ctr Chem & Biol Sci, Karachi 75270, Pakistan.
    Melittin Exerts Beneficial Effects on Paraquat-Induced Lung Injuries in Mice by Modifying Oxidative Stress and Apoptosis2019Ingår i: Molecules, ISSN 1431-5157, E-ISSN 1420-3049, Vol. 24, nr 8, artikel-id 1498Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Melittin (MEL) is a 26-amino acid peptide with numerous biological activities. Paraquat (PQ) is one of the most widely used herbicides, although it is extremely toxic to humans. To date, PQ poisoning has no effective treatment, and therefore the current study aimed to assess for the first time the possible effects of MEL on PQ-induced lung injuries in mice. Mice received a single intraperitoneal (IP) injection of PQ (30 mg/kg), followed by IP treatment with MEL (0.1 and 0.5 mg/kg) twice per week for four consecutive weeks. Histological alterations, oxidative stress, and apoptosis in the lungs were studied. Hematoxylin and eosin (H&E) staining indicated that MEL markedly reduced lung injuries induced by PQ. Furthermore, treatment with MEL increased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity, and decreased malonaldehyde (MDA) and nitric oxide (NO) levels in lung tissue homogenates. Moreover, immunohistochemical staining showed that B-cell lymphoma-2 (Bcl-2) and survivin expressions were upregulated after MEL treatment, while Ki-67 expression was downregulated. The high dose of MEL was more effective than the low dose in all experiments. In summary, MEL efficiently reduced PQ-induced lung injuries in mice. Specific pharmacological examinations are required to determine the effectiveness of MEL in cases of human PQ poisoning.

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  • 347.
    Elf, Johan
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för cell- och molekylärbiologi, Molekylär systembiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Hypothesis: Homologous Recombination Depends on Parallel Search2016Ingår i: CELL SYSTEMS, ISSN 2405-4712, Vol. 3, nr 4, s. 325-327Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    It is not known how a cell manages to find a specific DNA sequence sufficiently fast to repair a broken chromosome through homologous recombination. I propose a solution based on freely diffusing molecules that are programmed with sequences corresponding to those flanking the break site. In such a process, the target search would be parallelized.

  • 348.
    Elias, Mikael H.
    et al.
    Univ Minnesota, Dept Biochem Mol Biol & Biophys, St Paul, MN 55108 USA.;Univ Minnesota, BioTechnol Inst, St Paul, MN 55108 USA..
    Fraser, James S.
    Univ Calif San Francisco, Dept Bioengn & Therapeut Sci, San Francisco, CA 94143 USA..
    Kamerlin, Shina C. Lynn
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för kemi - BMC, Biokemi.
    Patrick, Wayne M.
    Victoria Univ Wellington, Sch Biol Sci, Wellington, New Zealand..
    Jackson, Colin J.
    Australian Natl Univ, Res Sch Chem, Canberra, ACT, Australia..
    Biographical item: Dan Salah Tawfik (1955-2021): A giant of protein evolution. In: EMBO Reports, Volume 22, Issue 7, Article Number e53256, 20212021Övrigt (Övrigt vetenskapligt)
  • 349.
    Elksnis, Andris
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Pharmaceutical Protection of Beta-Cells in Diabetes: Using Tyrosine Kinase Inhibition and NOX4 Inhibitors2022Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Diabetes mellitus is a complex and heterogenous disease, with loss of beta-cell function and mass being a characteristic of not only type 1 diabetes (T1D), but also type 2 diabetes (T2D). In T1D, inappropriate inflammatory signaling is thought to participate in the autoimmune suppression and destruction of beta-cells. In T2D progressive insulin resistance with resulting glucolipotoxicity, increased inflammation and oxidative stress, drives islet amyloid formation and subsequent beta-cell exhaustion and failure. Even under best managed care, disease progression and eventual complications are unavoidable. New interventions that aim to improve beta-cell survival are highly needed. This thesis investigates two such possible interventions: the tyrosine kinase inhibitor Imatinib, and selective NADPH-oxidase inhibition.

    Imatinib mesylate, used in treatment of chronic myeloid leukemia and other malignancies, was soon after its introduction reported to possess anti-diabetic properties in both T1D and T2D patients undergoing treatment. Imatinib has been shown to prevent and reverse diabetes in NOD mice and improve glucose tolerance in high fat diet treated rats. In paper I, we aimed to characterize the mechanisms by which imatinib protects beta-cells. We found that imatinib inhibits complex I and II of the respiratory chain, leading to improved beta-cell survival through AMPK activation, reduced amyloid formation and protection against TXNIP upregulation.

    Oxidative stress may play a pivotal role in the development of beta-cell dysfunction and failure in T2D. The NADPH-oxidases are a family of 7 enzymes (NOX1-5 and DUOX 1-2), that produce reactive oxygen species that are important in various physiological processes but may, if excessively activated, also be a source for oxidative stress in T2D. In paper II, we evaluate novel selective NOX inhibitors as protective agents against in vitro induced human beta-cell stress. Selective NOX4 inhibition protected beta-cells against both cytokines and high-glucose + palmitate. In paper III we found that NOX4 inhibition increased mitochondrial membrane potential, mitochondrial reactive oxygen species and ATP/ADP ratio in a human beta-cell line, and this was paralleled with protection against human islet cell death when challenged with high-glucose and palmitate. Finally, in paper IV, we attempt to apply these findings in vivo, by transplanting athymic diabetic mice with human islets and treating them with a NOX4 inhibitor over a period of 4 weeks. Treated mice achieved lower blood glucose levels and water consumption throughout the treatment period, and apoptotic rates of insulin-positive human cells, measured as co-localization of insulin and cleaved caspase-3, were greatly reduced.

    Delarbeten
    1. Imatinib protects against human beta-cell death via inhibition of mitochondrial respiration and activation of AMPK
    Öppna denna publikation i ny flik eller fönster >>Imatinib protects against human beta-cell death via inhibition of mitochondrial respiration and activation of AMPK
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    2021 (Engelska)Ingår i: Clinical Science, ISSN 0143-5221, E-ISSN 1470-8736, Vol. 135, nr 19, s. 2243-2263Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The protein tyrosine kinase inhibitor imatinib is used in the treatment of various malignancies but may also promote beneficial effects in the treatment of diabetes. The aim of the present investigation was to characterize the mechanisms by which imatinib protects insulin producing cells. Treatment of non-obese diabetic (NOD) mice with imatinib resulted in increased beta-cell AMP-activated kinase (AMPK) phosphorylation. Imatinib activated AMPK also in vitro, resulting in decreased ribosomal protein S6 phosphorylation and protection against islet amyloid polypeptide (IAPP)-aggregation, thioredoxin interacting protein (TXNIP) up-regulation and beta-cell death. 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) mimicked and compound C counteracted the effect of imatinib on beta-cell survival. Imatinib-induced AMPK activation was preceded by reduced glucose/pyruvate-dependent respiration, increased glycolysis rates, and a lowered ATP/AMP ratio. Imatinib augmented the fractional oxidation of fatty acids/malate, possibly via a direct interaction with the beta-oxidation enzyme enoyl coenzyme A hydratase, short chain, 1, mitochondrial (ECHS1). In non-beta cells, imatinib reduced respiratory chain complex I and II-mediated respiration and acyl-CoA carboxylase (ACC) phosphorylation, suggesting that mitochondrial effects of imatinib are not beta-cell specific. In conclusion, tyrosine kinase inhibitors modestly inhibit mitochondrial respiration, leading to AMPK activation and TXNIP down-regulation, which in turn protects against beta-cell death.

    Ort, förlag, år, upplaga, sidor
    Portland PressPORTLAND PRESS LTD, 2021
    Nationell ämneskategori
    Biokemi och molekylärbiologi Endokrinologi och diabetes
    Identifikatorer
    urn:nbn:se:uu:diva-458241 (URN)10.1042/CS20210604 (DOI)000707198700002 ()34569605 (PubMedID)
    Forskningsfinansiär
    BarndiabetesfondenDiabetesförbundetStiftelsen familjen Ernfors fondEXODIAB - Excellence of Diabetes Research in Sweden
    Tillgänglig från: 2021-11-16 Skapad: 2021-11-16 Senast uppdaterad: 2024-01-15Bibliografiskt granskad
    2. The novel NADPH oxidase 4 selective inhibitor GLX7013114 counteracts human islet cell death in vitro
    Öppna denna publikation i ny flik eller fönster >>The novel NADPH oxidase 4 selective inhibitor GLX7013114 counteracts human islet cell death in vitro
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    2018 (Engelska)Ingår i: PLOS ONE, E-ISSN 1932-6203, Vol. 13, nr 9, artikel-id e0204271Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    It has been proposed that pancreatic beta-cell dysfunction in type 2 diabetes is promoted by oxidative stress caused by NADPH oxidase (Nox) over-activity. The aim of the present study was to evaluate the efficacy of novel Nox inhibitors as protective agents against cytokine- or high glucose + palmitate-induced human beta-cell death. The Nox2 protein was present mainly in the cytoplasm and was induced by cytokines. Nox4 protein immunoreactivity, with some nuclear accumulation, was observed in human islet cells, and was not affected by islet culture in the presence of cytokines or high glucose + palmitate. Nox inhibitors with partial or no isoform selectivity (DPI, dapsone, GLX351322, and GLX481372) all reduced ROS production of human islet cells exposed to high glucose + palmitate. This was paralleled by improved viability and reduced caspase 3 activation. The Nox1 selective inhibitor ML171 failed to reduce human islet cell death in response to both cytokines and high glucose + palmitate. The selective Nox2 inhibitor Phox-12 also failed to protect against cytokines, but protected partially against high glucose + palmitate-induced cellular death. The highly selective Nox4 inhibitor GLX7013114 protected islet cells against both cytokines and high glucose + palmitate. However, as no osmotic control for high glucose was used, we cannot exclude the possibility that the high glucose effect was due to osmosis. It is concluded that Nox4 may participate in stress-induced islet cell death in human islets in vitro. We propose that Nox4 mediates pro-apoptotic effects in intact islets under stressful conditions and that selective Nox4-inhibition may be a therapeutic strategy in type 2 diabetes.

    Ort, förlag, år, upplaga, sidor
    PUBLIC LIBRARY SCIENCE, 2018
    Nationell ämneskategori
    Cell- och molekylärbiologi
    Identifikatorer
    urn:nbn:se:uu:diva-368106 (URN)10.1371/journal.pone.0204271 (DOI)000446005500024 ()30265686 (PubMedID)
    Forskningsfinansiär
    Vetenskapsrådet, 2017-01343Stiftelsen Olle Engkvist ByggmästareDiabetesförbundetNovo NordiskStiftelsen familjen Ernfors fondBarndiabetesfonden
    Tillgänglig från: 2018-12-03 Skapad: 2018-12-03 Senast uppdaterad: 2022-04-11Bibliografiskt granskad
    3. Pharmacological Inhibition of NOX4 Improves Mitochondrial Function and Survival in Human Beta-Cells
    Öppna denna publikation i ny flik eller fönster >>Pharmacological Inhibition of NOX4 Improves Mitochondrial Function and Survival in Human Beta-Cells
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    2021 (Engelska)Ingår i: Biomedicines, E-ISSN 2227-9059, Vol. 9, nr 12, artikel-id 1865Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Previous studies have reported beneficial effects of NADPH oxidase 4 (NOX4) inhibition on beta-cell survival in vitro and in vivo. The mechanisms by which NOX4 inhibition protects insulin producing cells are, however, not known. The aim of the present study was to investigate the effects of a pharmacological NOX4 inhibitor (GLX7013114) on human islet and EndoC-beta H1 cell mitochondrial function, and to correlate such effects with survival in islets of different size, activity, and glucose-stimulated insulin release responsiveness. We found that maximal oxygen consumption rates, but not the rates of acidification and proton leak, were increased in islets after acute NOX4 inhibition. In EndoC-beta H1 cells, NOX4 inhibition increased the mitochondrial membrane potential, as estimated by JC-1 fluorescence; mitochondrial reactive oxygen species (ROS) production, as estimated by MitoSOX fluorescence; and the ATP/ADP ratio, as assessed by a bioluminescent assay. Moreover, the insulin release from EndoC-beta H1 cells at a high glucose concentration increased with NOX4 inhibition. These findings were paralleled by NOX4 inhibition-induced protection against human islet cell death when challenged with high glucose and sodium palmitate. The NOX4 inhibitor protected equally well islets of different size, activity, and glucose responsiveness. We conclude that pharmacological alleviation of NOX4-induced inhibition of beta-cell mitochondria leads to increased, and not decreased, mitochondrial ROS, and this was associated with protection against cell death occurring in different types of heterogeneous islets. Thus, NOX4 inhibition or modulation may be a therapeutic strategy in type 2 diabetes that targets all types of islets.

    Ort, förlag, år, upplaga, sidor
    MDPIMDPI AG, 2021
    Nyckelord
    NOX4, beta-cell, mitochondria, cell death
    Nationell ämneskategori
    Endokrinologi och diabetes Cell- och molekylärbiologi
    Identifikatorer
    urn:nbn:se:uu:diva-465071 (URN)10.3390/biomedicines9121865 (DOI)000736240800001 ()34944680 (PubMedID)
    Forskningsfinansiär
    Vetenskapsrådet, 2017-01343Olle Engkvists stiftelseDiabetesförbundetNovo NordiskStiftelsen familjen Ernfors fondBarndiabetesfondenEXODIAB - Excellence of Diabetes Research in Sweden
    Tillgänglig från: 2022-01-21 Skapad: 2022-01-21 Senast uppdaterad: 2024-01-15Bibliografiskt granskad
    4. The selective NOX4 inhibitor GLX7013159 decreases blood glucose concentrations and human beta-cell apoptotic rates in diabetic NMRI nu/nu mice transplanted with human islets
    Öppna denna publikation i ny flik eller fönster >>The selective NOX4 inhibitor GLX7013159 decreases blood glucose concentrations and human beta-cell apoptotic rates in diabetic NMRI nu/nu mice transplanted with human islets
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    2023 (Engelska)Ingår i: Free radical research, ISSN 1071-5762, E-ISSN 1029-2470, Vol. 57, nr 6-12, s. 460-469Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    NADPH oxidase 4 (NOX4) inhibition has been reported to mitigate diabetes-induced beta-cell dysfunction and improve survival in vitro, as well as counteract high-fat diet-induced glucose intolerance in mice. We investigated the antidiabetic effects of the selective NOX4 inhibitor GLX7013159 in vivo in athymic diabetic mice transplanted with human islets over a period of 4 weeks. The GLX7013159-treated mice achieved lower blood glucose and water consumption throughout the treatment period. Furthermore, GLX7013159 treatment resulted in improved insulin and c-peptide levels, better insulin secretion capacity, as well as in greatly reduced apoptotic rates of the insulin-positive human cells, measured as colocalization of insulin and cleaved caspase-3. We conclude that the antidiabetic effects of NOX4 inhibition by GLX7013159 are observed also during a prolonged study period in vivo and are likely to be due to an improved survival and function of the human beta-cells.

    Ort, förlag, år, upplaga, sidor
    Taylor & Francis, 2023
    Nyckelord
    NOX inhibitor, NOX4, NADPH oxidase, reactive oxygen radical, human islet, beta-cell death, diabetes
    Nationell ämneskategori
    Endokrinologi och diabetes Cell- och molekylärbiologi
    Identifikatorer
    urn:nbn:se:uu:diva-472195 (URN)10.1080/10715762.2023.2284637 (DOI)001118087200001 ()
    Forskningsfinansiär
    Diabetesfonden, DIA-2020-540Vetenskapsrådet, 72XD-15043Stiftelsen familjen Ernfors fondBarndiabetesfondenEXODIAB - Excellence of Diabetes Research in SwedenOlle Engkvists stiftelse
    Tillgänglig från: 2022-04-07 Skapad: 2022-04-07 Senast uppdaterad: 2024-02-20Bibliografiskt granskad
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  • 350.
    Elksnis, Andris
    et al.
    Uppsala universitet, Science for Life Laboratory, SciLifeLab. Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi.
    Cen, Jing
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Wikström, Per
    Glucox Biotech AB, Fralsegardsvagen 8, SE-17997 Farentuna, Sweden..
    Carlsson, Per-Ola
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi. Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Transplantation och regenerativ medicin. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Welsh, Nils
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk cellbiologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Pharmacological Inhibition of NOX4 Improves Mitochondrial Function and Survival in Human Beta-Cells2021Ingår i: Biomedicines, E-ISSN 2227-9059, Vol. 9, nr 12, artikel-id 1865Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Previous studies have reported beneficial effects of NADPH oxidase 4 (NOX4) inhibition on beta-cell survival in vitro and in vivo. The mechanisms by which NOX4 inhibition protects insulin producing cells are, however, not known. The aim of the present study was to investigate the effects of a pharmacological NOX4 inhibitor (GLX7013114) on human islet and EndoC-beta H1 cell mitochondrial function, and to correlate such effects with survival in islets of different size, activity, and glucose-stimulated insulin release responsiveness. We found that maximal oxygen consumption rates, but not the rates of acidification and proton leak, were increased in islets after acute NOX4 inhibition. In EndoC-beta H1 cells, NOX4 inhibition increased the mitochondrial membrane potential, as estimated by JC-1 fluorescence; mitochondrial reactive oxygen species (ROS) production, as estimated by MitoSOX fluorescence; and the ATP/ADP ratio, as assessed by a bioluminescent assay. Moreover, the insulin release from EndoC-beta H1 cells at a high glucose concentration increased with NOX4 inhibition. These findings were paralleled by NOX4 inhibition-induced protection against human islet cell death when challenged with high glucose and sodium palmitate. The NOX4 inhibitor protected equally well islets of different size, activity, and glucose responsiveness. We conclude that pharmacological alleviation of NOX4-induced inhibition of beta-cell mitochondria leads to increased, and not decreased, mitochondrial ROS, and this was associated with protection against cell death occurring in different types of heterogeneous islets. Thus, NOX4 inhibition or modulation may be a therapeutic strategy in type 2 diabetes that targets all types of islets.

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