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Sökning: WFRF:(Jonsson Anders) > Göteborgs universitet

  • Resultat 11-20 av 110
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11.
  • Sandstedt, Mikael, 1990, et al. (författare)
  • Intracellular flow cytometry may be combined with good quality and high sensitivity RT-qPCR analysis.
  • 2015
  • Ingår i: Cytometry. Part A : the journal of the International Society for Analytical Cytology. - : Wiley. - 1552-4930. ; 87:12
  • Tidskriftsartikel (refereegranskat)abstract
    • Flow cytometry (FCM) has become a well-established method for analysis of both intracellular and cell-surface proteins, while quantitative RT-PCR (RT-qPCR) is used to determine gene expression with high sensitivity and specificity. Combining these two methods would be of great value. The effects of intracellular staining on RNA integrity and RT-qPCR sensitivity and quality have not, however, been fully examined. We, therefore, intended to assess these effects further. Cells from the human lung cancer cell line A549 were fixed, permeabilized and sorted by FCM. Sorted cells were analyzed using RT-qPCR. RNA integrity was determined by RNA quality indicator analysis. A549 cells were then mixed with cells of the mouse cardiomyocyte cell line HL-1. A549 cells were identified by the cell surface marker ABCG2, while HL-1 cells were identified by intracellular cTnT. Cells were sorted and analyzed by RT-qPCR. Finally, cell cultures from human atrial biopsies were used to evaluate the effects of fixation and permeabilization on RT-qPCR analysis of nonimmortalized cells stored prior to analysis by FCM. A large amount of RNA could be extracted even when cells had been fixed and permeabilized. Permeabilization resulted in increased RNA degradation and a moderate decrease in RT-qPCR sensitivity. Gene expression levels were also affected to a moderate extent. Sorted populations from the mixed A549 and HL-1 cell samples showed gene expression patterns that corresponded to FCM data. When samples were stored before FCM sorting, the RT-qPCR analysis could still be performed with high sensitivity and quality. In summary, our results show that intracellular FCM may be performed with only minor impairment of the RT-qPCR sensitivity and quality when analyzing sorted cells; however, these effects should be considered when comparing RT-qPCR data of not fixed samples with those of fixed and permeabilized samples. © 2015 International Society for Advancement of Cytometry.
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12.
  • Sandstedt, Mikael, 1990, et al. (författare)
  • Regional transcriptomic profiling reveals immune system enrichment in nonfailing atria as well as all chambers of the failing human heart
  • 2023
  • Ingår i: American Journal of Physiology. Heart and Circulatory Physiology. - : American Physiological Society. - 0363-6135 .- 1522-1539. ; 325:6, s. H1430-H1445
  • Tidskriftsartikel (refereegranskat)abstract
    • The different chambers of the human heart demonstrate regional physiological traits and may be differentially affected during pathologic remodeling, resulting in heart failure. Few previous studies have, however, characterized the different chambers at a transcriptomic level. We therefore conducted whole-tissue RNA sequencing and gene set enrichment analysis of biopsies collected from the four chambers of adult failing (n = 8) and nonfailing (n = 11) human hearts. Atria and ventricles demonstrated distinct transcriptional patterns. Compared to nonfailing ventricles, the transcriptional pattern of nonfailing atria was enriched for a large number of gene sets associated with cardiogenesis, the immune system and bone morphogenetic protein (BMP), transforming growth factor beta (TGF beta), MAPK/JNK and Wnt signaling. Differences between failing and nonfailing hearts were also determined. The transcriptional pattern of failing atria was distinct compared to that of nonfailing atria and enriched for gene sets associated with the innate and adaptive immune system, TGF beta/SMAD signaling, and changes in endothelial, smooth muscle cell and cardiomyocyte physiology. Failing ventricles were also enriched for gene sets associated with the immune system. Based on the transcriptomic patterns, upstream regulators associated with heart failure were identified. These included many immune response factors predicted to be similarly activated for all chambers of failing hearts. In summary, the heart chambers demonstrate distinct transcriptional patterns that differ between failing and nonfailing hearts. Immune system signaling may be a hallmark of all four heart chambers in failing hearts, and could constitute a novel therapeutic target.
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13.
  • Sjölin, Jacob, et al. (författare)
  • Expression of Stem Cell Niche-Related Biomarkers at the Base of the Human Tricuspid Valve
  • 2023
  • Ingår i: Stem Cells and Development. - : Mary Ann Liebert Inc. - 1547-3287 .- 1557-8534. ; 32:5-6, s. 140-151
  • Tidskriftsartikel (refereegranskat)abstract
    • Stem cell niches have been thoroughly investigated in tissue with high regenerative capacity but not in tissues where cell turnover is slow, such as the human heart. The left AtrioVentricular junction (AVj), the base of the mitral valve, has previously been proposed as a niche region for cardiac progenitors in the adult human heart. In the present study, we explore the right side of the human heart, the base of the tricuspid valve, to investigate the potential of this region as a progenitor niche. Paired biopsies from explanted human hearts were collected from multi-organ donors (N = 12). The lateral side of the AVj, right atria (RA), and right ventricle (RV) were compared for the expression of stem cell niche-related biomarkers using RNA sequencing. Gene expression data indicated upregulation of genes related to embryonic development and extracellular matrix (ECM) composition in the proposed niche region, that is, the AVj. In addition, immunohistochemistry showed high expression of the fetal cardiac markers MDR1, SSEA4, and WT1 within the same region. Nuclear expression of HIF1 alpha was detected suggesting hypoxia. Rare cells were found with the co-staining of the proliferation marker PCNA and Ki67 with cardiomyocyte nuclei marker PCM1 and cardiac Troponin T (cTnT), indicating proliferation of small cardiomyocytes. WT1+/cTnT+ and SSEA4+/cTnT+ cells were also found, suggesting cardiomyocyte-specific progenitors. The expression of the stem cell markers gradually decreased with distance from the tricuspid valve. No expression of these markers was observed in the RV tissue. In summary, the base of the tricuspid valve is an ECM-rich region containing cells with expression of several stem cell niche-associated markers. Co-expression of stem cell markers with cTnT indicates cardiomyocyte-specific progenitors. We previously reported similar data from the base of the mitral valve and thus propose that human adult cardiomyocyte progenitors reside around both atrioventricular valves.
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14.
  • Synnergren, Jane, et al. (författare)
  • Transcriptional sex and regional differences in paired human atrial and ventricular cardiac biopsies collected in vivo
  • 2020
  • Ingår i: Physiological Genomics. - : American Physiological Society. - 1094-8341 .- 1531-2267. ; 52:2, s. 110-120
  • Tidskriftsartikel (refereegranskat)abstract
    • Transcriptional studies of the human heart provide insight into physiological and pathophysiological mechanisms, essential for understanding the fundamental mechanisms of normal cardiac function and how they are altered by disease. To improve the understanding of why men and women may respond differently to the same therapeutic treatment it is crucial to learn more about sex-specific transcriptional differences. In this study the transcriptome of right atrium and left ventricle was compared across sex and regional location. Paired biopsies from five male and five female patients undergoing aortic valve replacement or coronary artery bypass grafting were included. Gene expression analysis identified 620 differentially expressed transcripts in atrial and ventricular tissue in men and 471 differentially expressed transcripts in women. In total 339 of these transcripts overlapped across sex but notably, 281 were unique in the male tissue and 162 in the female tissue, displaying marked sex differences in the transcriptional machinery. The transcriptional activity was significantly higher in atrias than in ventricles as 70% of the differentially expressed genes were upregulated in the atrial tissue. Furthermore, pathway- and functional annotation analyses performed on the differentially expressed genes showed enrichment for a more heterogeneous composition of biological processes in atrial compared with the ventricular tissue, and a dominance of differentially expressed genes associated with infection disease was observed. The results reported here provide increased insights about transcriptional differences between the cardiac atrium and ventricle but also reveal transcriptional differences in the human heart that can be attributed to sex.
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15.
  • Vukusic, Kristina, 1979, et al. (författare)
  • High Density Sphere Culture of Adult Cardiac Cells Increases the Levels of Cardiac and Progenitor Markers and Shows Signs of Vasculogenesis
  • 2013
  • Ingår i: Biomed Research International. - : Hindawi Limited. - 2314-6133 .- 2314-6141.
  • Tidskriftsartikel (refereegranskat)abstract
    • 3D environment and high cell density play an important role in restoring and supporting the phenotypes of cells represented in cardiac tissues.. e aim of this study was therefore to investigate the suitability of high density sphere (HDS) cultures for studies of cardiomyocyte-, endothelial-, and stem-cell biology. Primary adult cardiac cells from nine human biopsies were cultured using different media for up to 9 weeks.. e possibilities to favor a certain cell phenotype and induce production of extra cellular matrix (ECM) were studied by histology, immunohistochemistry, and uantitative real-time PCR. Defined media gave significant increase in both cardiac-and progenitor-specific markers and also an intraluminal position of endothelial cells over time. Cardiac media showed indication of differentiation and maturity of HDS considering the ECM production and activities within NOTCH regulation but no additional cardiac differentiation. Endothelial media gave no positive effects on endothelial phenotype but increased proliferation without fibroblast overgrowth. In addition, indications for early vasculogenesis were found. It was also possible to affect the Wnt signaling in HDS by addition of a glycogen synthase kinase 3 (GSK3) inhibitor. In conclusion, these findings show the suitability of HDS as in vitro model for studies of cardiomyocyte-, endothelial-, and stem-cell biology.
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16.
  • Allwood, Carl Martin, 1952, et al. (författare)
  • Child witnesses meta-memory realism
  • 2006
  • Ingår i: Scandinavian Journal of Psychology. - 0036-5564 .- 1467-9450. ; 47, s. 461-470
  • Tidskriftsartikel (refereegranskat)
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17.
  • Allwood, Carl Martin, 1952, et al. (författare)
  • Child witnesses’ metamemory realism
  • 2006
  • Ingår i: Scandinavian Journal of Psychology. - : Wiley. - 0036-5564 .- 1467-9450. ; 47, s. 461-470
  • Tidskriftsartikel (refereegranskat)abstract
    • This study investigated the degree of realism in the confidence judgments of 11-12 year old children (N=81) of their answers to questions relating to a short film clip showing a kidnapping event. Four different confidence scales were used: a numeric scale, a picture scale, a line scale, and a written scale. The results demonstrated that the children showed a high level of overconfidence in their memories. However, no significant differences between the four confidence scales were found. The results indicate that, at least in the context investigated, 11-12 year-old children’s confidence in their event memory show poor realism. A comparison with previous research on adults indicates that children show noticeably poorer realism.
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18.
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19.
  • Allwood, Carl Martin, 1952, et al. (författare)
  • The effects of source and type of feedback on child witnesses' metamemory accuracy
  • 2005
  • Ingår i: Applied Cognitive Psychology. - : Wiley. - 0888-4080 .- 1099-0720. ; 19:3, s. 331-344
  • Tidskriftsartikel (refereegranskat)abstract
    • This study investigated the effect of feedback on the accuracy (realism) of 12-year-old children's metacognitive judgments of their answers to questions about a film clip. Two types of judgments were investigated: confidence judgments (on each question) and frequency judgments (i.e. estimates of overall accuracy). The source of feedback, whether it was presented as provided by a teacher or a peer child, did not influence metacognitive accuracy. Four types of feedback were given depending on whether the participant's answer was correct and depending on whether the feedback confirmed or disconfirmed the child's answer. The children showed large overconfidence when they received confirmatory feedback but much less so when they received disconfirmatory feedback. The children gave frequency judgments implying that they had more correct answers than they actually had. No main gender differences were found for any of the measures. The results indicate a high degree of malleability in children's metacognitive judgments.
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20.
  • Andersson, Malte, 1941, et al. (författare)
  • ”Minskande befolkning är inte problemet”
  • 2020
  • Ingår i: Dagens Nyheter. ; :1 augusti, DN-debatt
  • Tidskriftsartikel (populärvet., debatt m.m.)abstract
    • Nätverket Population Matters Sweden: En uppmärksammad studie i The Lancet pekar mot en lägre befolkningsökning i världen än tidigare prognoser. Men en miljard människor till är fortfarande långt över vad jorden klarar. Befolkningstrenden måste snarare vända neråt, och det kräver åtgärder för att stärka kvinnors rättigheter världen över.
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