217 research outputs found
Miro Remo, The Film Author
Get PDFDizertační práce se komplexně zabývá filmovou tvorbou slovenského režiséra Mira Rema. Výzkum vycházející ze stěžejní úvahy o Removi jakožto tvůrci s osobitým autorským rukopisem stojí na několika opěrných bodech. V jedné z rovin se zaměřuje na témata a postavy, kterými se Remo ve svých filmech zabývá, a s ohledem na to reflektuje, skrze jaké režijní metody, stylové a narativní postupy či prvky filmové řeči je zpřítomňuje filmovému publiku (včetně toho, do jaké míry se opakují či variují). Vedle takto zaměřených analýz jednotlivých děl je pozornost koncentrována i na okolnosti jejich vzniku a dílčí fáze jejich produkce, tzn. na vývoj, výrobu, postprodukci a distribuci. Velký prostor je rovněž věnován kritickému diskurzu a na základě zevrubných rešerší je mapováno, jakým způsobem se projekty Mira Rema diskutovaly v médiích, jaké kauzy vyvolávaly nebo jak rezonovaly u filmové kritiky a v rámci festivalové scény. Výzkum zahrnuje i sérii polostrukturovaných rozhovorů: jak se samotným Mirem Remem, tak i s jeho blízkými spolupracovníky ze všech hlavních štábních profesí ? scenáristou a dramaturgem Jurem Šlaukou, kreativním producentem Markem Kučerou, kameramanem Jarem Vaľkem, střihačem Markem Kráľovským a zvukařem Lukášem Kasprzykem.The thesis deals comprehensively with the film work of the Slovak director Miro Remo. The research is based on the central reflection of Remo as a film author who possesses distinctive handwriting. On one level, it focuses on the themes and characters that Remo deals with. The theses also reflects on the directorial methods, stylistic and narrative approaches or elements of film language through which the director communicates them to the film audience (including the extent to which they are repeated or varied). In addition to such focused analyses of individual works, attention is also given to the circumstances of of their creation as well as the sub-phases of their production, i.e. their development, preproduction, shooting, post-production and distribution. A great deal of the work is also dedicated to critical discourse and ? based on extensive research ? it is mapped how Remo's projects were discussed in the media, what kind of controversies they provoked or how they resonated with film critics or within the festival scene. The research includes a series of semi-structured interviews: with Miro Remo himself, as well as with his close collaborators from several professions ? screenwriter and script consultant Juro Šlauka, creative producer Marek Kučera, DOP Jaro Vaľko, editor Marek Kráľovský and sound designer Lukáš Kasprzyk.Ateliér Audioviz
The VP1u of Human Parvovirus B19: A Multifunctional Capsid Protein with Biotechnological Applications
Get PDFThe viral protein 1 unique region (VP1u) of human parvovirus B19 (B19V) is a multifunctional capsid protein with essential roles in virus tropism, uptake, and subcellular trafficking. These functions reside on hidden protein domains, which become accessible upon interaction with cell membrane receptors. A receptor-binding domain (RBD) in VP1u is responsible for the specific targeting and uptake of the virus exclusively into cells of the erythroid lineage in the bone marrow. A phospholipase A2 domain promotes the endosomal escape of the incoming virus. The VP1u is also the immunodominant region of the capsid as it is the target of neutralizing antibodies. For all these reasons, the VP1u has raised great interest in antiviral research and vaccinology. Besides the essential functions in B19V infection, the remarkable erythroid specificity of the VP1u makes it a unique erythroid cell surface biomarker. Moreover, the demonstrated capacity of the VP1u to deliver diverse cargo specifically to cells around the proerythroblast differentiation stage, including erythroleukemic cells, offers novel therapeutic opportunities for erythroid-specific drug delivery. In this review, we focus on the multifunctional role of the VP1u in B19V infection and explore its potential in diagnostics and erythroid-specific therapeutics
Urban models and public-private partnership
No full textThis book addresses the topic of urban models with reference to large western cities and particularly to global cities. In the current transitional phase, the use of language and the systematization of phenomena has become important. The book’s matrix examines two important and strongly connected themes: urban models and public-private partnerships (PPP) determined by urban functions which are transformed in an increasingly rapid and complex manner as a result of globalization. PPPs represent the new border of the modern global state. The book focuses on two principal urban models (renewal and restructuring) through PPPs and subsequently the relationship between state and market in fourteen Italian cities (renewal) and two central European cities, Leipzig and Budapest (restructuring). CoUrbIT (Complex Urban Investment Tools) and the book 'Globalization and Urban Implosion: Creating New Competitive Advantage' by the same author serve as points of referenc
Scepsi e mito in Remo Cantoni
Get PDFSkepticism and Myth in Remo Cantoni. The author examines three essays by Remo Cantoni (1914-1978), written in the 1940s, which focus on the relationship between skepticism and myth. The author argues that, for Cantoni, skepticism and myth represent two fundamental polarities of thought. The hypothesis that reason might ultimately overcome myth is, according to Cantoni, erroneous. Instead, the author emphasizes that Cantoni considers a phenomenology of myth essential for understanding it as a necessary dimension in the life of the spirit. According to the author, Cantoni is a philosopher who seeks to interpret the crisis of values and science, striving to remain within a rationalistic framework
Scepsi e mito in Remo Cantoni
No full textSkepticism and Myth in Remo Cantoni. The author examines three essays by Remo Cantoni (1914-1978), written in the 1940s, which focus on the relationship between skepticism and myth. The author argues that, for Cantoni, skepticism and myth represent two fundamental polarities of thought.The hypothesis that reason might ultimately overcome myth is, according to Cantoni, erroneous. Instead, the author emphasizes that Cantoni considers a phenomenology of myth essential for understanding it as a necessary dimension in the life of the spirit.According to the author, Cantoni is a philosopher who seeks to interpret the crisis of values and science, striving to remain within a rationalistic framework
Scepsi e mito in Remo Cantoni
No full textSkepticism and Myth in Remo Cantoni. The author examines three essays by Remo Cantoni (1914-1978), written in the 1940s, which focus on the relationship between skepticism and myth. The author argues that, for Cantoni, skepticism and myth represent two fundamental polarities of thought. The hypothesis that reason might ultimately overcome myth is, according to Cantoni, erroneous. Instead, the author emphasizes that Cantoni considers a phenomenology of myth essential for understanding it as a necessary dimension in the life of the spirit. According to the author, Cantoni is a philosopher who seeks to interpret the crisis of values and science, striving to remain within a rationalistic framework
The VP1u Receptor Restricts Parvovirus B19 Uptake to Permissive Erythroid Cells
Get PDFParvovirus B19 (B19V) is a small non-enveloped virus and known as the causative agent for the mild childhood disease erythema infectiosum. B19V has an extraordinary narrow tissue tropism, showing only productive infection in erythroid precursor cells in the bone marrow. We recently found that the viral protein 1 unique region (VP1u) contains an N-terminal receptor-binding domain (RBD), which mediates the uptake of the virus into cells of the erythroid lineage. To further investigate the role of the RBD in connection with a B19V-unrelated capsid, we chemically coupled the VP1u of B19V to the bacteriophage MS2 capsid and tested the internalization capacity of the bioconjugate on permissive cells. In comparison, we studied the cellular uptake and infection of B19V along the erythroid differentiation. The results showed that the MS2-VP1u bioconjugate mimicked the specific internalization of the native B19V into erythroid precursor cells, which further coincides with the restricted infection profile. The successful mimicry of B19V uptake demonstrates that the RBD in the VP1u is sufficient for the endocytosis of the viral capsid. Furthermore, the recombinant VP1u competed with B19V uptake into permissive cells, thus excluding a significant alternative uptake mechanism by other receptors. Strikingly, the VP1u receptor appeared to be expressed only on erythropoietin-dependent erythroid differentiation stages that also provide the necessary intracellular factors for a productive infection. Taken together, these findings suggest that the VP1u binds to a yet-unknown erythroid-specific cellular receptor and thus restricts the virus entry to permissive cells
The Receptor-Binding Domain in the VP1u Region of Parvovirus B19
Get PDFParvovirus B19 (B19V) is known as the human pathogen causing the mild childhood disease erythema infectiosum. B19V shows an extraordinary narrow tissue tropism for erythroid progenitor cells in the bone marrow, which is determined by a highly restricted uptake. We have previously shown that the specific internalization is mediated by the interaction of the viral protein 1 unique region (VP1u) with a yet unknown cellular receptor. To locate the receptor-binding domain (RBD) within the VP1u, we analyzed the effect of truncations and mutations on the internalization capacity of the recombinant protein into UT7/Epo cells. Here we report that the N-terminal amino acids 5-80 of the VP1u are necessary and sufficient for cellular binding and internalization; thus, this N-terminal region represents the RBD required for B19V uptake. Using site-directed mutagenesis, we further identified a cluster of important amino acids playing a critical role in VP1u internalization. In silico predictions and experimental results suggest that the RBD is structured as a rigid fold of three α-helices. Finally, we found that dimerization of the VP1u leads to a considerably enhanced cellular binding and internalization. Taken together, we identified the RBD that mediates B19V uptake and mapped functional and structural motifs within this sequence. The findings reveal insights into the uptake process of B19V, which contribute to understand the pathogenesis of the infection and the neutralization of the virus by the immune system
Human parvovirus B19 interacts with globoside under acidic conditions as an essential step in endocytic trafficking
Get PDFThe glycosphingolipid (GSL) globoside (Gb4) is essential for parvovirus B19 (B19V) infection. Historically considered the cellular receptor of B19V, the role of Gb4 and its interaction with B19V are controversial. In this study, we applied artificial viral particles, genetically modified cells, and specific competitors to address the interplay between the virus and the GSL. Our findings demonstrate that Gb4 is not involved in the binding or internalization process of the virus into permissive erythroid cells, a function that corresponds to the VP1u cognate receptor. However, Gb4 is essential at a post-internalization step before the delivery of the single-stranded viral DNA into the nucleus. In susceptible erythroid Gb4 knockout cells, incoming viruses were arrested in the endosomal compartment, showing no cytoplasmic spreading of capsids as observed in Gb4-expressing cells. Hemagglutination and binding assays revealed that pH acts as a switch to modulate the affinity between the virus and the GSL. Capsids interact with Gb4 exclusively under acidic conditions and dissociate at neutral pH. Inducing a specific Gb4-mediated attachment to permissive erythroid cells by acidification of the extracellular environment led to a non-infectious uptake of the virus, indicating that low pH-mediated binding to the GSL initiates active membrane processes resulting in vesicle formation. In summary, this study provides mechanistic insight into the interaction of B19V with Gb4. The strict pH-dependent binding to the ubiquitously expressed GSL prevents the redirection of the virus to nonpermissive tissues while promoting the interaction in acidic intracellular compartments as an essential step in infectious endocytic trafficking
Determination of parvovirus retention profiles in virus filter membranes using laser scanning microscopy
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