Saccharomyces cerevisiae Rio2p (encoded by open reading frame Ynl207w) is an essential protein of unknown function that displays significant sequence similarity to Rio1p/Rrp10p. The latter was recently shown to be an evolutionarily conserved, predominantly cytoplasmic serine/threonine kinase whose presence is required for the final cleavage at site D that converts 20 S pre-rRNA into mature 18 S rRNA. A data base search identified homologs of Rio2p in a wide variety of eukaryotes and Archaea. Detailed sequence comparison and in vitro kinase assays using recombinant protein demonstrated that Rio2p defines a subfamily of protein kinases related to, but both structurally and functionally distinct from, the one defined by Rio1p. Failure to deplete Rio2p in cells containing a GAL-rio2 gene and direct analysis of Rio2p levels by Western blotting indicated the protein to be low abundant. Using a GAL-rio2 gene carrying a point mutation that reduces the kinase activity, we found that depletion of this mutant protein blocked production of 18 S rRNA due to inhibition of the cleavage of cytoplasmic 20 S pre-rRNA at site D. Production of the large subunit rRNAs was not affected. Thus, Rio2p is the second protein kinase that is essential for cleavage at site D and the first in which the processing defect can be linked to its enzymatic activity. Contrary to Rio1p/Rrp10p, however, Rio2p appears to be localized predominantly in the nucleus.Like their counterparts in other eukaryotes, Saccharomyces cerevisiae ribosomes contain four species of rRNA: 5 S, 5.8 S, 18 S, and 25 S rRNAs. The genes encoding these rRNAs are organized on the yeast genome in 150 -200 tandem repeats, each of which comprises two transcriptional units separated by non-transcribed spacers. One of these units consists of a 5 S rRNA gene, transcribed by RNA polymerase III. The other unit contains single genes for each of the mature 18 S, 5.8 S, and 25 S rRNAs that are separated by internal transcribed spacers 1 and 2, whereas external transcribed spacer regions are present at either end of the unit (see Fig. 1A). After transcription of this polycistronic unit by RNA polymerase I, the spacers are removed from the primary transcript via an ordered series of endo-and exonucleolytic cleavages (see Fig. 1B) (reviewed in Refs. 1 and 2). The first detectable precursor species is 35 S pre-rRNA, which results from a cleavage at site B 0 in the 3Ј-external transcribed spacer by the yeast RNase III homolog Rnt1p (3,4). Subsequent cleavage at sites A 0 and A 1 in the 5Ј-external transcribed spacer first gives rise to 33 S and then 32 S pre-rRNA. The latter is cleaved at site A 2 to produce separate 20 S and 27 S A 2 precursors for the small and large ribosomal subunit, respectively.The majority (90%) of the 27 S A 2 precursor molecules are cleaved endonucleolytically at site A 3 , followed by exonucleolytic trimming to B 1S . The remainder are processed endonucleolytically 1 at site B 1L . The resulting 27 S B S and 27 S B L precursors, whose 5Ј-ends are located 6 nucleotide...
This article challenges the assumption that patient autonomy can best be assured by providing proper information through formalized procedures such as informed consent. We suggest that to understand and consider laypeople's ways of knowing and decision making, one has to move beyond the information paradigm and take into account a much broader context. Concretely, we investigate informed consent in connection with donating skin tissue remaining from medically indicated surgery. We use interviews with patients and observation protocols to analyse patients' perceptions and ways of making sense of informed consent beyond its bioethical ideal. Patients situate themselves in a larger system of solidarity, enroll in an overall positive image of science as a linear process of innovation oriented towards output, and simultaneously take a pragmatic stance towards hospital routines as a necessary passage point towards receiving good treatment. Because informed consent is one of the central articulations between the biomedical system and society, we conclude by reflecting on the consequences of our findings on a socio-political level.
Melanoma is one of the most aggressive neoplastic transformations and characterized by a high metastatic potential. The current study was performed to assess the impact of "spleen tyrosine kinase" (Syk), a non-receptor-associated tyrosine kinase, on growth and metastatic behavior of melanoma cells in vitro and in a severe combined immunodeficient (SCID)-mouse/human-melanoma xenotransplantation model in vivo. Syk was expressed in melanocytes but was found to be downregulated in melanoma cells. Vector-driven expression of Syk in two different melanoma cell lines did not influence growth speed, but significantly reduced the invasive growth potential of both cell lines in a Matrigel assay in vitro. In a SCID-mouse/human melanoma xenotransplantation model, Syk expressing Mel-Juso cells exhibited delayed and reduced tumor growth. After intravenous as well as subcutaneous injection of tumor cells, Syk-transfected cells formed significantly fewer metastatic tumor lesions than control cells. The presented data define Syk as a novel regulator of metastatic behavior of melanoma cells.
In ethnographic research and analysis, reflexivity is vital to achieving constant coordination between field and concept work. However, it has been conceptualized predominantly as an ethnographer’s individual mental capacity. In this article, we draw on ten years of experience in conducting research together with partners from social psychiatry and mental health care across different research projects. We unfold three modes of achieving reflexivity co-laboratively: contrasting and discussing disciplinary concepts in interdisciplinary working groups and feedback workshops; joint data interpretation and writing; and participating in political agenda setting. Engaging these modes reveals reflexivity as a distributed process able to strengthen the ethnographer’s interpretative authority, and also able to constantly push the conceptual boundaries of the participating disciplines and professions.
For almost half a century social scientists have explored the phenomenon of chronic illness. In this paper, I examine how the concept of chronicity participates in present-day mental health care settings. Using ethnomethodology and material-semiotic theory within science and technology studies, I investigate how the classification 'chronically mentally ill' interacts with the everyday socio-material shaping of public mental health care in the context of professional institutions. Drawing on ethnographic fieldwork in a psychiatric day hospital and in a community day care centre in Berlin, Germany, I demonstrate how the classification of chronicity acts as a tool of description (of people or their conditions), regulation (of therapy, health care or administration), and connection to infrastructures of care (practised technologies or standards of various kinds). In these ways, I argue, the classification engages in actions of producing treatability, arranging resources, demarcating responsibilities, practicing accountability, and doing presence. Notably, community mental health care has developed into a designated territory of the concept: explicitly arranged for 'the chronically mentally ill' as a human kind, we can take everyday life in these institutions as instructive of how chronicity is defined in daily practice.
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