11 research outputs found
Constitutional Law - First Amendment - New York Statute Proscribing Distribution of Nonobscene Materials Depicting Minors Engaged in Sexual Conduct Does Not Violate the First Amendment because the Materials Are Outside First Amendment Protection and the Statute Is Not Substantially Overboard
Constitutional Law - First Amendment - New York Statute Proscribing Distribution of Nonobscene Materials Depicting Minors Engaged in Sexual Conduct Does Not Violate the First Amendment because the Materials Are Outside First Amendment Protection and the Statute Is Not Substantially Overboard
Appropriately differentiated ARPE-19 cells regain phenotype and gene expression profiles similar to those of native RPE cells.
PurposeThe RPE cell line ARPE-19 provides a dependable and widely used alternative to native RPE. However, replication of the native RPE phenotype becomes more difficult because these cells lose their specialized phenotype after multiple passages. Compounding this problem is the widespread use of ARPE-19 cells in an undifferentiated state to attempt to model RPE functions. We wished to determine whether suitable culture conditions and differentiation could restore the RPE-appropriate expression of genes and proteins to ARPE-19, along with a functional and morphological phenotype resembling native RPE. We compared the transcriptome of ARPE-19 cells kept in long-term culture with those of primary and other human RPE cells to assess the former's inherent plasticity relative to the latter.MethodsARPE-19 cells at passages 9 to 12 grown in DMEM containing high glucose and pyruvate with 1% fetal bovine serum were differentiated for up to 4 months. Immunocytochemistry was performed on ARPE-19 cells grown on filters. Total RNA extracted from ARPE-19 cells cultured for either 4 days or 4 months was used for RNA sequencing (RNA-Seq) analysis using a 2 × 50 bp paired end protocol. The RNA-Seq data were analyzed to identify the affected pathways and recognize shared ontological classification among differentially expressed genes. RPE-specific mRNAs and miRNAs were assessed with quantitative real-time (RT)-PCR, and proteins with western blotting.ResultsARPE-19 cells grown for 4 months developed the classic native RPE phenotype with heavy pigmentation. RPE-expressed genes, including RPE65, RDH5, and RDH10, as well as miR-204/211, were greatly increased in the ARPE-19 cells maintained at confluence for 4 months. The RNA-Seq analysis provided a comprehensive view of the relative abundance and differential expression of the genes in the differentiated ARPE-19 cells. Of the 16,757 genes with detectable signals, nearly 1,681 genes were upregulated, and 1,629 genes were downregulated with a fold change of 2.5 or more differences between 4 months and 4 days of culture. Gene Ontology analysis showed that the upregulated genes were associated with visual cycle, phagocytosis, pigment synthesis, cell differentiation, and RPE-related transcription factors. The majority of the downregulated genes play a role in cell cycle and proliferation.ConclusionsThe ARPE-19 cells cultured for 4 months developed a phenotype characteristic of native RPE and expressed proteins, mRNAs, and miRNAs characteristic of the RPE. Comparison of the ARPE-19 RNA-Seq data set with that of primary human fetal RPE, embryonic stem cell-derived RPE, and native RPE revealed an important overall similar expression ratio among all the models and native tissue. However, none of the cultured models reached the absolute values in the native tissue. The results of this study demonstrate that low-passage ARPE-19 cells can express genes specific to native human RPE cells when appropriately cultured and differentiated
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Expression of ABCA4 in the retinal pigment epithelium and its implications for Stargardt macular degeneration.
Recessive Stargardt disease (STGD1) is an inherited blinding disorder caused by mutations in the Abca4 gene. ABCA4 is a flippase in photoreceptor outer segments (OS) that translocates retinaldehyde conjugated to phosphatidylethanolamine across OS disc membranes. Loss of ABCA4 in Abca4 -/- mice and STGD1 patients causes buildup of lipofuscin in the retinal pigment epithelium (RPE) and degeneration of photoreceptors, leading to blindness. No effective treatment currently exists for STGD1. Here we show by several approaches that ABCA4 is additionally expressed in RPE cells. (i) By in situ hybridization analysis and by RNA-sequencing analysis, we show the Abca4 mRNA is expressed in human and mouse RPE cells. (ii) By quantitative immunoblotting, we show that the level of ABCA4 protein in homogenates of wild-type mouse RPE is about 1% of the level in neural retina homogenates. (iii) ABCA4 immunofluorescence is present in RPE cells of wild-type and Mertk -/- but not Abca4 -/- mouse retina sections, where it colocalizes with endolysosomal proteins. To elucidate the role of ABCA4 in RPE cells, we generated a line of genetically modified mice that express ABCA4 in RPE cells but not in photoreceptors. Mice from this line on the Abca4 -/- background showed partial rescue of photoreceptor degeneration and decreased lipofuscin accumulation compared with nontransgenic Abca4 -/- mice. We propose that ABCA4 functions to recycle retinaldehyde released during proteolysis of rhodopsin in RPE endolysosomes following daily phagocytosis of distal photoreceptor OS. ABCA4 deficiency in the RPE may play a role in the pathogenesis of STGD1
Harmful algal blooms and eutrophication : examining linkages from selected coastal regions of the United States
Author Posting. © Elsevier B.V., 2008. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Harmful Algae 8 (2008): 39-53, doi:10.1016/j.hal.2008.08.017.Coastal waters of the United States (U.S.) are subject to many of the major harmful algal
bloom (HAB) poisoning syndromes and impacts. These include paralytic shellfish poisoning
(PSP), neurotoxic shellfish poisoning (NSP), amnesic shellfish poisoning (ASP), ciguatera
fish poisoning (CFP) and various other HAB phenomena such as fish kills, loss of submerged
vegetation, shellfish mortalities, and widespread marine mammal mortalities. Here, the
occurrences of selected HABs in a selected set of regions are described in terms of their
relationship to eutrophication, illustrating a range of responses. Evidence suggestive of
changes in the frequency, extent or magnitude of HABs in these areas is explored in the
context of the nutrient sources underlying those blooms, both natural and anthropogenic. In
some regions of the U.S., the linkages between HABs and eutrophication are clear and well
documented, whereas in others, information is limited, thereby highlighting important areas
for further research.Support was provided through the Woods Hole Center for Oceans
and Human Health (to DMA), National Science Foundation (NSF) grants OCE-9808173 and
OCE-0430724 (to DMA), OCE-0234587 (to WPC), OCE04-32479 (to MLP), OCE-0138544
(to RMK), OCE-9981617 (to PMG); National Institute of Environmental Health Sciences
(NIEHS) grants P50ES012742-01 (to DMA) and P50ES012740 (to MLP); NOAA Grants
NA96OP0099 (to DMA), NA16OP1450 (to VLT), NA96P00084 (to GAV and CAH),
NA160C2936 and NA108H-C (to RMK), NA860P0493 and NA04NOS4780241 (to PMG),
NA04NOS4780239-02 (to RMK), NA06NOS4780245 (to DWT). Support was also provided from the West Coast Center for Oceans and Human Health (to VLT and WPC), USEPA
Grant CR826792-01-0 (to GAV and CAH), and the State of Florida Grant S7701617826 (to
GAV and CAH)
Part of a candidate disease region (AXPC1) showing a novel spliced gene transcript
This view shows a detail of one end of the AXPC1 region. An expressed sequence tag (EST) from the retinal pigment epithelium (RPE) shows the structure (vertical bars are exons; arrowed lines show introns) of a novel gene. EyeSAGE has tag counts for some Unigenes in this region, including the novel gene (designated as C1orf132 in Unigene). Note that the EyeSAGE tag bars are positioned according to the corresponding Unigene. Their positions may change slightly with different releases of Unigene.<p><b>Copyright information:</b></p><p>Taken from "NEIBank: Genomics and bioinformatics resources for vision research"</p><p></p><p>Molecular Vision 2008;14():1327-1337.</p><p>Published online 18 Jul 2008</p><p>PMCID:PMC2480482.</p><p></p
Expression of ABCA4 in the retinal pigment epithelium and its implications for Stargardt macular degeneration
As abordagens market-driven e market-driving de orientação para o mercado e inovação: proposição de um modelo integrado Market-driving strategy for market orientation: a theoretical model and suggestions for research
Direcionamentos recentes da teoria de orientação para o mercado têm apontado para a natureza eminentemente reativa das estratégias relacionadas. Argumenta-se que a abordagem tradicional - market-driven - seja excessivamente determinista e de forte conotação adaptativa perante o ambiente externo. Essa visão passa a ser complementada por uma nova perspectiva - market-driving -, pressupondo a possibilidade de que as estruturas e o comportamento do mercado possam ser modelados pelas organizações. Entretanto, ainda são incipientes os esforços na construção de um modelo de referência que possa nortear futuras pesquisas no âmbito dessa estratégia. É nessa problemática que se insere o objetivo do presente trabalho, em vistas de apresentar um modelo teórico e proposições de pesquisa sobre o tema, baseados em intensiva revisão de literatura. Como conclusão, o modelo construído permite vislumbrar os antecedentes da estratégia market-driving, suas respectivas ações de implementação e variáveis correlatas. Também se verifica a natureza complementar dessa abordagem em relação à postura tradicional de orientação. Ao final, apresentam-se as implicações acadêmicas e gerenciais do presente estudo, bem como a agenda para pesquisas futuras.<br>Recent developments in the market orientation theory have been indicating for the reactive nature of the related strategies. It argues that the traditional approach, market-driven, is excessively determinism and of strong adaptative connotation in front of external atmosphere. That vision becomes complemented by a new perspective, market-driving, presupposing that the market structures and its behavior can be modeled by the organizations. However, this theory is still incipient in the construction of a reference model that can orientate future research using that strategy. This paper looks for fulfill this gap by presenting a theoretical model and some research propositions on the theme, based on intensive literature review. As conclusion, the model allows to analyze the antecedents of the market-driving strategy, review some actions to be implemented and verify correlated variables. The complementally nature of that approach is also verified in relation to the traditional posture of orientation. At the end, some academic and managerial implications as discussed, as well as, the agenda for future research