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  1. Article ; Online: In Vitro Methods to Study the Golgi Apparatus Role in Proteoglycan and Glycosaminoglycan Synthesis.

    Prydz, Kristian

    Methods in molecular biology (Clifton, N.J.)

    2023  Volume 2557, Page(s) 709–720

    Abstract: Subcellular fractionation is an introductory step in a variety of experimental approaches designed to study intracellular components, like membranes and organelle systems. Subcellular fractions enriched in membranes of the Golgi apparatus of mammalian ... ...

    Abstract Subcellular fractionation is an introductory step in a variety of experimental approaches designed to study intracellular components, like membranes and organelle systems. Subcellular fractions enriched in membranes of the Golgi apparatus of mammalian cells have been isolated to address localization and activity of proteins, including enzymes, to study intracellular membrane transport mechanisms, and to reconstitute in vitro cellular processes associated with the Golgi apparatus. Here, I describe methods to purify Golgi membranes by subcellular fractionation, to assay nucleotide sulfate (PAPS) uptake into Golgi vesicles, and to measure sulfate incorporation into in vitro synthesized glycosaminoglycans.
    MeSH term(s) Animals ; Phosphoadenosine Phosphosulfate/metabolism ; Proteoglycans/metabolism ; Golgi Apparatus/metabolism ; Glycosaminoglycans/metabolism ; Sulfates/metabolism ; Mammals/metabolism
    Chemical Substances Phosphoadenosine Phosphosulfate (482-67-7) ; Proteoglycans ; Glycosaminoglycans ; Sulfates
    Language English
    Publishing date 2023-02-15
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2639-9_42
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  2. Article ; Online: The life cycle and enigmatic egress of coronaviruses.

    Prydz, Kristian / Saraste, Jaakko

    Molecular microbiology

    2022  Volume 117, Issue 6, Page(s) 1308–1316

    Abstract: There has been considerable recent interest in the life cycle of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), the causative agent of the Covid-19 pandemic. Practically every step in CoV replication-from cell attachment and uptake via ... ...

    Abstract There has been considerable recent interest in the life cycle of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), the causative agent of the Covid-19 pandemic. Practically every step in CoV replication-from cell attachment and uptake via genome replication and expression to virion assembly has been considered as a specific event that potentially could be targeted by existing or novel drugs. Interference with cellular egress of progeny viruses could also be adopted as a possible therapeutic strategy; however, the situation is complicated by the fact that there is no broad consensus on how CoVs find their way out of their host cells. The viral nucleocapsid, consisting of the genomic RNA complexed with nucleocapsid proteins obtains a membrane envelope during virus budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface. From here, several alternative routes for CoV extracellular release have been proposed. Strikingly, recent studies have shown that CoV infection leads to the disassembly of the Golgi ribbon and the mobilization of host cell compartments and protein machineries that are known to promote Golgi-independent trafficking to the cell surface. Here, we discuss the life cycle of CoVs with a special focus on different possible pathways for virus egress.
    MeSH term(s) Animals ; COVID-19 ; Humans ; Life Cycle Stages ; Pandemics ; SARS-CoV-2 ; Viral Envelope Proteins/genetics
    Chemical Substances Viral Envelope Proteins
    Language English
    Publishing date 2022-05-04
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 619315-8
    ISSN 1365-2958 ; 0950-382X
    ISSN (online) 1365-2958
    ISSN 0950-382X
    DOI 10.1111/mmi.14907
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    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Metabolic Labeling of Proteoglycans and Analysis of Their Synthesis and Sorting in Filter-Grown and Polarized Epithelial Cells.

    Prydz, Kristian / Adusumalli, Ravi

    Methods in molecular biology (Clifton, N.J.)

    2021  Volume 2303, Page(s) 25–36

    Abstract: Studies of synthesis, turnover, and secretion of macromolecules in cell culture are carried out to address mechanisms of cellular and physiological importance. Culture systems have been developed to mimic the in vivo situation as much as possible. In ... ...

    Abstract Studies of synthesis, turnover, and secretion of macromolecules in cell culture are carried out to address mechanisms of cellular and physiological importance. Culture systems have been developed to mimic the in vivo situation as much as possible. In line with this aim, epithelial and endothelial cells have been grown on filters for more than three decades. Growing such cells on permeable support allows for nutrient uptake via the basolateral membrane of tight epithelial monolayers, from a medium reservoir underneath the filter. While this basolateral medium reservoir resembles the blood supply, the apical medium reservoir resembles the organ lumen. Growing the cells in a polarized manner allows for studies of differential transport and localization of apical and basolateral proteins and of endocytic and secretory transport at both sides of the epithelium. Here we describe how metabolic labeling of proteoglycans (PGs) with
    MeSH term(s) Cell Line ; Endothelial Cells/metabolism ; Epithelial Cells/metabolism ; Kidney/metabolism ; Protein Transport ; Proteoglycans/metabolism
    Chemical Substances Proteoglycans
    Language English
    Publishing date 2021-10-08
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-1398-6_3
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  4. Article ; Online: Assembly and Cellular Exit of Coronaviruses: Hijacking an Unconventional Secretory Pathway from the Pre-Golgi Intermediate Compartment via the Golgi Ribbon to the Extracellular Space.

    Saraste, Jaakko / Prydz, Kristian

    Cells

    2021  Volume 10, Issue 3

    Abstract: Coronaviruses (CoVs) assemble by budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface. However, why CoVs have chosen the IC as their intracellular site of assembly and how progeny viruses are ... ...

    Abstract Coronaviruses (CoVs) assemble by budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface. However, why CoVs have chosen the IC as their intracellular site of assembly and how progeny viruses are delivered from this compartment to the extracellular space has remained unclear. Here we address these enigmatic late events of the CoV life cycle in light of recently described properties of the IC. Of particular interest are the emerging spatial and functional connections between IC elements and recycling endosomes (REs), defined by the GTPases Rab1 and Rab11, respectively. The establishment of IC-RE links at the cell periphery, around the centrosome and evidently also at the noncompact zones of the Golgi ribbon indicates that-besides traditional ER-Golgi communication-the IC also promotes a secretory process that bypasses the Golgi stacks, but involves its direct connection with the endocytic recycling system. The initial confinement of CoVs to the lumen of IC-derived large transport carriers and their preferential absence from Golgi stacks is consistent with the idea that they exit cells following such an unconventional route. In fact, CoVs may share this pathway with other intracellularly budding viruses, lipoproteins, procollagen, and/or protein aggregates experimentally introduced into the IC lumen.
    MeSH term(s) Animals ; COVID-19/therapy ; COVID-19/virology ; Centrosome/metabolism ; Endoplasmic Reticulum/virology ; Extracellular Space/metabolism ; Extracellular Space/virology ; Golgi Apparatus/metabolism ; Golgi Apparatus/virology ; Humans ; Intracellular Membranes/virology ; Protein Transport ; SARS-CoV-2/physiology ; Secretory Pathway ; Virus Release
    Language English
    Publishing date 2021-02-26
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells10030503
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  5. Article ; Online: Collecting evidence of validity for an assessment tool for Norwegian medical students' non-technical skills (NorMS-NTS): usability and reliability when used by novice raters.

    Prydz, Katrine / Dieckmann, Peter / Fagertun, Hans / Musson, David / Wisborg, Torben

    BMC medical education

    2023  Volume 23, Issue 1, Page(s) 865

    Abstract: Background: The NorMS-NTS tool is an assessment tool for assessing Norwegian medical students' non-technical skills (NTS). The NorMS-NTS was designed to provide student feedback, training evaluations, and skill-level comparisons among students at ... ...

    Abstract Background: The NorMS-NTS tool is an assessment tool for assessing Norwegian medical students' non-technical skills (NTS). The NorMS-NTS was designed to provide student feedback, training evaluations, and skill-level comparisons among students at different study sites. Rather than requiring extensive rater training, the tool should capably suit the needs of busy doctors as near-peer educators. The aim of this study was to examine the usability and preliminary assess validity of the NorMS-NTS tool when used by novice raters.
    Methods: This study focused on the usability of the assessment tool and its internal structure. Three raters used the NorMS-NTS tool to individually rate the team leader, a medical student, in 20 video-recorded multi-professional simulation-based team trainings. Based on these ratings, we examined the tools' internal structure by calculating the intraclass correlation coefficient (ICC) (version 3.1) interrater reliability, internal consistency, and observability. After the rating process was completed, the raters answered a questionnaire about the tool's usability.
    Results: The ICC agreement and the sum of the overall global scores for all raters were fair: ICC (3,1) = 0.53. The correlation coefficients for the pooled raters were in the range of 0.77-0.91. Cronbach's alpha for elements, categories and global score were mostly above 0.90. The observability was high (95%-100%). All the raters found the tool easy to use, none of the elements were redundant, and the written instructions were helpful. The raters also found the tool easier to use once they had acclimated to it. All the raters stated that they could use the tool for both training and teaching.
    Conclusions: The observed ICC agreement was 0.08 below the suggested ICC level for formative assessment (above 0.60). However, we know that the suggestion is based on the average ICC, which is always higher than a single-measure ICC. There are currently no suggested levels for single-measure ICC, but other validated NTS tools have single-measure ICC in the same range. We consider NorMS-NTS as a usable tool for formative assessment of Norwegian medical students' non-technical skills during multi-professional team training by raters who are new to the tool. It is necessary to further examine validity and the consequences of the tool to fully validate it for formative assessments.
    MeSH term(s) Humans ; Students, Medical ; Reproducibility of Results ; Physicians ; Educational Measurement ; Feedback ; Clinical Competence
    Language English
    Publishing date 2023-11-15
    Publishing country England
    Document type Journal Article
    ZDB-ID 2044473-4
    ISSN 1472-6920 ; 1472-6920
    ISSN (online) 1472-6920
    ISSN 1472-6920
    DOI 10.1186/s12909-023-04837-6
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  6. Article ; Online: Determinants of Glycosaminoglycan (GAG) Structure.

    Prydz, Kristian

    Biomolecules

    2015  Volume 5, Issue 3, Page(s) 2003–2022

    Abstract: Proteoglycans (PGs) are glycosylated proteins of biological importance at cell surfaces, in the extracellular matrix, and in the circulation. PGs are produced and modified by glycosaminoglycan (GAG) chains in the secretory pathway of animal cells. The ... ...

    Abstract Proteoglycans (PGs) are glycosylated proteins of biological importance at cell surfaces, in the extracellular matrix, and in the circulation. PGs are produced and modified by glycosaminoglycan (GAG) chains in the secretory pathway of animal cells. The most common GAG attachment site is a serine residue followed by a glycine (-ser-gly-), from which a linker tetrasaccharide extends and may continue as a heparan sulfate, a heparin, a chondroitin sulfate, or a dermatan sulfate GAG chain. Which type of GAG chain becomes attached to the linker tetrasaccharide is influenced by the structure of the protein core, modifications occurring to the linker tetrasaccharide itself, and the biochemical environment of the Golgi apparatus, where GAG polymerization and modification by sulfation and epimerization take place. The same cell type may produce different GAG chains that vary, depending on the extent of epimerization and sulfation. However, it is not known to what extent these differences are caused by compartmental segregation of protein cores en route through the secretory pathway or by differential recruitment of modifying enzymes during synthesis of different PGs. The topic of this review is how different aspects of protein structure, cellular biochemistry, and compartmentalization may influence GAG synthesis.
    MeSH term(s) Animals ; Glycosaminoglycans/chemistry ; Glycosaminoglycans/metabolism ; Golgi Apparatus/metabolism ; Humans ; Membrane Transport Proteins/metabolism ; Secretory Pathway
    Chemical Substances Glycosaminoglycans ; Membrane Transport Proteins
    Language English
    Publishing date 2015-08-21
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2701262-1
    ISSN 2218-273X ; 2218-273X
    ISSN (online) 2218-273X
    ISSN 2218-273X
    DOI 10.3390/biom5032003
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  7. Article ; Online: The development of a tool to assess medical students' non-technical skills - The Norwegian medical students' non-technical skills (NorMS-NTS).

    Prydz, Katrine / Dieckmann, Peter / Musson, David / Wisborg, Torben

    Medical teacher

    2022  Volume 45, Issue 5, Page(s) 516–523

    Abstract: Purpose: New physicians need to master non-technical skills (NTS), as high levels of NTS have been shown to increase patient safety. It has also been shown that NTS can be improved through training. This study aimed to establish the necessary NTS for ... ...

    Abstract Purpose: New physicians need to master non-technical skills (NTS), as high levels of NTS have been shown to increase patient safety. It has also been shown that NTS can be improved through training. This study aimed to establish the necessary NTS for Norwegian medical students to create a tool for formative and summative assessments.
    Methods: Focus group interviews were conducted with colleagues and patients of newly graduated physicians. Interviews were then analyzed using card sort methods, and the identified NTS were used to establish a framework. Focus groups commented on a prototype of an NTS assessment tool. Finally, we conducted a search of existing tools and literature. The final tool was developed based on the combined inputs.
    Results: We created Norwegian medical students' non-technical skills (NorMS-NTS) assessment tool containing four main categories; together comprising 13 elements and a rating scale for the NTS of the person observed.
    Conclusions: The NorMS-NTS represents a purpose-made tool for assessing newly graduated physicians' NTS. It is similar to existing assessment tools but based on domain-specific user perspectives obtained through focus group interviews and feedback, integrated with results from a literature search, and with consideration of existing NTS tools.
    MeSH term(s) Humans ; Students, Medical ; Clinical Competence ; Physicians ; Focus Groups ; Norway
    Language English
    Publishing date 2022-11-07
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 424426-6
    ISSN 1466-187X ; 0142-159X
    ISSN (online) 1466-187X
    ISSN 0142-159X
    DOI 10.1080/0142159X.2022.2140034
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  8. Article: Specific Heats, C

    Prydz, Rolf / Goodwin, Robert D

    Journal of research of the National Bureau of Standards. Section A, Physics and chemistry

    2020  Volume 74A, Issue 5, Page(s) 661–665

    Abstract: ... reported from 80 K to 300 K at pressures to about 23 MN/m ...

    Abstract Experimental specific heats at constant volume for compressed gaseous and liquid fluorine are reported from 80 K to 300 K at pressures to about 23 MN/m
    Language English
    Publishing date 2020-08-03
    Publishing country United States
    Document type Journal Article
    ZDB-ID 241519-7
    ISSN 0022-4332
    ISSN 0022-4332
    DOI 10.6028/jres.074A.054
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    In stock of ZB MED Bonn / Germany

    Z 4' 62/62: Show issues

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  9. Article: Vitamin K--dependent clotting factors.

    Prydz, H

    Seminars in thrombosis and hemostasis

    1977  Volume 4, Issue 1, Page(s) 1–14

    MeSH term(s) Amino Acids ; Antithrombins/pharmacology ; Chemical Phenomena ; Chemistry ; Factor IX/biosynthesis ; Factor VII/biosynthesis ; Factor VII/metabolism ; Factor X/biosynthesis ; Glutamates/pharmacology ; Humans ; Prothrombin/biosynthesis ; Vitamin K/metabolism
    Chemical Substances Amino Acids ; Antithrombins ; Glutamates ; Vitamin K (12001-79-5) ; Factor VII (9001-25-6) ; Prothrombin (9001-26-7) ; Factor IX (9001-28-9) ; Factor X (9001-29-0)
    Language English
    Publishing date 1977
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 196901-8
    ISSN 1098-9064 ; 0094-6176
    ISSN (online) 1098-9064
    ISSN 0094-6176
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1259: Show issues Location:
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  10. Article ; Online: Evidence for the role of Rab11-positive recycling endosomes as intermediates in coronavirus egress from epithelial cells.

    Saraste, Jaakko / Enyioko, Mary / Dale, Hege / Prydz, Kristian / Machamer, Carolyn

    Histochemistry and cell biology

    2022  Volume 158, Issue 3, Page(s) 241–251

    Abstract: After their assembly by budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface, coronaviruses (CoVs) are released from their host cells following a pathway that remains poorly understood. The ... ...

    Abstract After their assembly by budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface, coronaviruses (CoVs) are released from their host cells following a pathway that remains poorly understood. The traditional view that CoV exit occurs via the constitutive secretory route has recently been questioned by studies suggesting that this process involves unconventional secretion. Here, using the avian infectious bronchitis virus (IBV) as a well-established model virus, we have applied confocal microscopy to investigate the pathway of CoV egress from epithelial Vero cells. We report a novel effect of IBV infection on cellular endomembranes, namely, the compaction of the pericentrosomal endocytic recycling compartment (ERC) defined by the GTPase Rab11, which coincides with the previously described Golgi fragmentation, as well as virus release. Despite Golgi disassembly, the IC elements containing the major IBV membrane protein (M)-which mostly associates with newly formed virus particles-maintain their close spatial connection with the Rab11-positive endocytic recycling system. Moreover, partial colocalization of the M protein with Rab11 was observed, whereas M displayed negligible overlap with LAMP-1, indicating that IBV egress does not occur via late endosomes or lysosomes. Synchronization of virus release using temperature-shift protocols was accompanied by increased colocalization of M and Rab11 in vesicular and vacuolar structures in the pericentrosomal region and at the cell periphery, most likely representing IBV-containing transport carriers. In conclusion, these results add CoVs to the growing list of viruses exploiting the endocytic recycling apparatus defined by Rab11 for their assembly and/or release.
    MeSH term(s) Animals ; Chlorocebus aethiops ; Coronavirus/metabolism ; Endosomes/metabolism ; Golgi Apparatus/metabolism ; Vero Cells ; rab GTP-Binding Proteins/metabolism
    Chemical Substances rab GTP-Binding Proteins (EC 3.6.5.2)
    Language English
    Publishing date 2022-05-23
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 1222930-1
    ISSN 1432-119X ; 0301-5564 ; 0948-6143
    ISSN (online) 1432-119X
    ISSN 0301-5564 ; 0948-6143
    DOI 10.1007/s00418-022-02115-y
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