LRP16 Integrates into NF-κB Transcriptional Complex and Is Required for Its Functional Activation

BACKGROUND: Nuclear factor κB (NF-κB)-mediated pathways have been widely implicated in cell survival, development and tumor progression. Although the molecular events of determining NF-κB translocation from cytoplasm to nucleus have been extensively documented, the regulatory mechanisms of NF-κB activity inside the nucleus are still poorly understood. Being a special member of macro domain proteins, LRP16 was previously identified as a coactivator of both estrogen receptor and androgen receptor, and as an interactor of NF-κB coactivator UXT. Here, we investigated the regulatory role of LRP16 on NF-κB activation. METHODOLOGY: GST pull-down and coimmunoprecipitation (CoIP) assays assessed protein-protein interactions. The functional activity of NF-κB was assessed by luciferase assays, changes in expression of its target genes, and its DNA binding ability. Annexin V staining and flow cytometry analysis were used to evaluate cell apoptosis. Immunohistochemical staining of LRP16 and enzyme-linked immunosorbent assay-based evaluation of active NF-κB were performed on primary human gastric carcinoma samples. RESULTS: We demonstrate that LRP16 integrates into NF-κB transcriptional complex through associating with its p65 component. RNA interference knockdown of the endogenous LRP16 in cells leads to impaired NF-κB activity and significantly attenuated NF-κB-dependent gene expression. Mechanistic analysis revealed that knockdown of LRP16 did not affect tumor necrosis factor α (TNF-α)-induced nuclear translocation of NF-κB, but blunted the formation or stabilization of functional NF-κB/p300/CREB-binding protein transcription complex in the nucleus. In addition, knockdown of LRP16 also sensitizes cells to apoptosis induced by TNF-α. Finally, a positive link between LRP16 expression intensity in nuclei of tumor cells and NF-κB activity was preliminarily established in human gastric carcinoma specimens. CONCLUSIONS: Our findings not only indicate that LRP16 is a crucial regulator for NF-κB activation inside the nucleus, but also suggest that LRP16 may be an important contributor to the aberrant activation of NF-κB in tumors

The impact of transposable element activity on therapeutically relevant human stem cells

Human stem cells harbor significant potential for basic and clinical translational research as well as regenerative medicine. Currently ~ 3000 adult and ~ 30 pluripotent stem cell-based, interventional clinical trials are ongoing worldwide, and numbers are increasing continuously. Although stem cells are promising cell sources to treat a wide range of human diseases, there are also concerns regarding potential risks associated with their clinical use, including genomic instability and tumorigenesis concerns. Thus, a deeper understanding of the factors and molecular mechanisms contributing to stem cell genome stability are a prerequisite to harnessing their therapeutic potential for degenerative diseases. Chemical and physical factors are known to influence the stability of stem cell genomes, together with random mutations and Copy Number Variants (CNVs) that accumulated in cultured human stem cells. Here we review the activity of endogenous transposable elements (TEs) in human multipotent and pluripotent stem cells, and the consequences of their mobility for genomic integrity and host gene expression. We describe transcriptional and post-transcriptional mechanisms antagonizing the spread of TEs in the human genome, and highlight those that are more prevalent in multipotent and pluripotent stem cells. Notably, TEs do not only represent a source of mutations/CNVs in genomes, but are also often harnessed as tools to engineer the stem cell genome; thus, we also describe and discuss the most widely applied transposon-based tools and highlight the most relevant areas of their biomedical applications in stem cells. Taken together, this review will contribute to the assessment of the risk that endogenous TE activity and the application of genetically engineered TEs constitute for the biosafety of stem cells to be used for substitutive and regenerative cell therapiesS.R.H. and P.T.R. are funded by the Government of Spain (MINECO, RYC-2016- 21395 and SAF2015–71589-P [S.R.H.]; PEJ-2014-A-31985 and SAF2015–71589- P [P.T.R.]). GGS is supported by a grant from the Ministry of Health of the Federal Republic of Germany (FKZ2518FSB403)

Preparo e administração venosa de medicamentos e soros sob a ótica da Resolução COFEN n° 311/07 Preparación y administración venosa de medicamentos y sueros bajo la óptica de la Resolución COFEN n° 311/07 Preparation and administration of intravenous drugs and serums from the perspective of Resolution No. 311/07 COFEN

OBJETIVOS: Levantar na literatura os procedimentos cabíveis ao enfermeiro quanto ao preparo e administração venosa de medicamentos e soros e relacioná-los com os aspectos éticos estabelecidos pela Resolução COFEN n° 311/07. MÉTODOS: Pesquisa do tipo revisão bibliográfica realizada nas bases de dados Medline, Lilacs, SciELO e Bdenf. Uma análise temática foi realizada em 13 artigos e emergiram as categorias: Procedimentos realizados pelo enfermeiro no preparo e na administração venosa; Responsabilidades éticas do enfermeiro: atuação do Poder Judiciário frente às decisões administrativas de natureza disciplinar proferidas pelo Conselho Federal de Enfermagem e Conselhos Regionais da Enfermagem. RESULTADOS: O enfermeiro deve realizar o preparo e administração venosa observando os requisitos básicos que garantam a ausência de danos ao cliente, e em conformidade com o Código de Ética dos profissionais de Enfermagem. CONCLUSÃO: O conhecimento da ética é essencial para a profissão da enfermagem, porque subsidia o embasamento teórico e propicia um fazer com resultados efetivos de forma a proteger o ser humano.<br>OBJETIVOS: Levantar en la literatura los procedimientos que le caben al enfermero en lo que se refiere a la preparación y administración venosa de medicamentos y sueros; y, relacionarlos con los aspectos éticos establecidos por la Resolución COFEN n° 311/07. MÉTODOS: Se trata de una investigación del tipo revisión bibliográfica, realizada en las bases de datos Medline, Lilacs, SciELO y Bdenf. Se realizó un análisis temático en 13 artículos de los que surgieron las categorías: 1) procedimientos realizados por el enfermero en la preparación y en la administración venosa; 2) responsabilidades éticas del enfermero (actuación del Poder Judicial frente a las decisiones administrativas de naturaleza disciplinar proferidas por el Consejo Federal de Enfermería y Consejos Regionales de la Enfermería). RESULTADOS: El enfermero debe realizar la preparación y administración venosa observando los requisitos básicos que garantizan la ausencia de daños al cliente, y de conformidad con el Código de Ética de los profesionales de Enfermería. CONCLUSIÓN: El conocimiento de la ética es esencial para la profesión de enfermería, porque subsidia el embasamiento teórico y es propicio a un quehacer con resultados efectivos de forma de proteger al ser humano.<br>OBJECTIVES: To survey in the literature the procedures performed by nurses related to the preparation and administration of intravenous medications and serums; and, to relate the procedures to ethical standards established by Resolution No. 311/07 COFEN. METHODS: The type of research was literature review, conducted in the databases Medline, Lilacs, SciELO and BDENF. A thematic analysis was performed on 13 items and two categories emerged: 1) procedures performed by nurses in preparing and administering intravenous medications; and 2) ethical responsibilities of nurses (the role of the judiciary in the face of administrative decisions of disciplinary nature made by the Federal Nursing Council and Regional Nursing Councils). RESULTS: The nurse must carry out the preparation and administration of intravenous medications in accordance with the basic requirements of ensuring no damage to the client, and also following the Code of Ethics of Professional Nursing. CONCLUSION: The knowledge of ethics is essential for the nursing profession because it subsidizes the theoretical foundations and provides the performing of tasks with effective results that have the intention of protecting the human being

Intrinsic epigenetic regulation of the D4Z4 macrosatellite repeat in a transgenic mouse model for FSHD

Contains fulltext : 118685.pdf (publisher's version ) (Open Access)Facioscapulohumeral dystrophy (FSHD) is a progressive muscular dystrophy caused by decreased epigenetic repression of the D4Z4 macrosatellite repeats and ectopic expression of DUX4, a retrogene encoding a germline transcription factor encoded in each repeat. Unaffected individuals generally have more than 10 repeats arrayed in the subtelomeric region of chromosome 4, whereas the most common form of FSHD (FSHD1) is caused by a contraction of the array to fewer than 10 repeats, associated with decreased epigenetic repression and variegated expression of DUX4 in skeletal muscle. We have generated transgenic mice carrying D4Z4 arrays from an FSHD1 allele and from a control allele. These mice recapitulate important epigenetic and DUX4 expression attributes seen in patients and controls, respectively, including high DUX4 expression levels in the germline, (incomplete) epigenetic repression in somatic tissue, and FSHD-specific variegated DUX4 expression in sporadic muscle nuclei associated with D4Z4 chromatin relaxation. In addition we show that DUX4 is able to activate similar functional gene groups in mouse muscle cells as it does in human muscle cells. These transgenic mice therefore represent a valuable animal model for FSHD and will be a useful resource to study the molecular mechanisms underlying FSHD and to test new therapeutic intervention strategies