BackgroundThe transcription factor NK2 homeobox 1 (Nkx2-1) plays essential roles in epithelial cell proliferation and differentiation in mouse and human lung development and tumorigenesis. A better understanding of genes and pathways downstream of Nkx2-1 will clarify the multiple roles of this critical lung factor. Nkx2-1 regulates directly or indirectly numerous protein-coding genes; however, there is a paucity of information about Nkx2-1-regulated microRNAs (miRNAs).Methods and resultsBy miRNA array analyses of mouse epithelial cell lines in which endogenous Nkx2-1 was knocked-down, we revealed that 29 miRNAs were negatively regulated including miR-200c, and 39 miRNAs were positively regulated by Nkx2-1 including miR-1195. Mouse lungs lacking functional phosphorylated Nkx2-1 showed increased expression of miR-200c and alterations in the expression of other top regulated miRNAs. Moreover, chromatin immunoprecipitation assays showed binding of NKX2-1 protein to regulatory regions of these miRNAs. Promoter reporter assays indicated that 1kb of the miR-200c 5′ flanking region was transcriptionally active but did not mediate Nkx2-1- repression of miR-200c expression. 3′UTR reporter assays support a direct regulation of the predicted targets Nfib and Myb by miR-200c.ConclusionsThese studies suggest that Nkx2-1 controls the expression of specific miRNAs in lung epithelial cells. In particular, we identified a regulatory link between Nkx2-1, the known tumor suppressor miR-200c, and the developmental and oncogenic transcription factors Nfib and Myb, adding new players to the regulatory mechanisms driven by Nkx2-1 in lung epithelial cells that may have implications in lung development and tumorigenesis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12931-015-0186-6) contains supplementary material, which is available to authorized users.

Edited by Jeffrey E. Pessin In mammals, leptin production in adipocytes is up-regulated by feeding and insulin. Although this regulatory connection is central to all physiological effects of leptin, its molecular mechanism remains unknown. Here, we show that the transcription factor early growth response 1, Egr1, is rapidly but transiently induced by insulin in adipose cells both in vitro and in vivo, and its induction is followed by an increase in leptin transcription. ChIP and luciferase assays demonstrate that Egr1 directly binds to and activates the leptin promoter. Interestingly, the lipid droplet protein FSP27 may work as a co-factor for Egr1 in regulating leptin expression. By using siRNA-mediated knockout of Egr1 along with its overexpression in adipocytes, we demonstrate that Egr1 is both necessary and sufficient for the stimulatory effect of insulin on leptin transcription.

The vasculopathy of sickle cell disease (SCD), characterized by a chronic hemolytic anemia, endothelial dysfunction and recurrent vaso-occlusive events, underlies most of the clinical complications. Hydroxyurea (HU), the only FDA approved medication to treat patients with SCD, acts via the induction of fetal hemoglobin (HbF) production. Recent studies suggest that HU increases endothelial nitric oxide (NO) production and reduces adhesion molecule expression as another potential mechanism for the observed reduction in vaso-occlusive events and improved survival in SCD. Despite its benefits, HU is currently under-utilized and we hypothesize that improved targeted drug delivery may increase its usage. The use of nano-emulsions, a class of stable emulsions formed by a monolayer of phospholipids and/or biodegradable material, is a mechanism by which to enhance targeted cellular delivery of therapeutics. We hypothesized that a nano-formulation of HU would enhance drug delivery to the bone marrow and this may be a direct way of impacting HbF expression in SCD. Nano-formulated HU (NHU) was developed using a phospholipid approach and this compound was found to be biologically stable for > 6 months. Chronic myelogenous leukemia bone marrow (K562) cells were treated with a NHU or HU suspension in concentrations ranging from 25µM to 250µM over a course of 4, 5 and 6 days. Cells were harvested for RNA isolation and qPCR was performed using primers for HbF and basal cell adhesion molecule (BCAM, a cellular adhesion molecule on red and white blood cells. On day 5, when HbF expression in K562 cells peaks, there was a five-fold increase in HbF expression treated with lower concentrations of NHU compared to untreated cells and a two-fold increase in expression compared with HU treated cells. With increasing concentrations of NHU and HU, HbF expression became similar. This suggests that the use of NHU can provide increased HbF expression at a lower concentration with likely less toxicity. Treatment of K562 cells with NHU and HU resulted in decreased BCAM expression on day 6 of treatment. Treatment with NHU produced a four-fold decrease in BCAM expression compared to the same concentration of a HU suspension which supports the concept that NHU can impact gene expression at lower doses than unformulated HU. Nanoformulated HU produces a molecule which is highly stable, and able to achieve higher intracellular concentrations with increased efficacy in K562 cells. This suggests a potential novel therapeutic for patients with sickle cell disease. Disclosures Klings: Pfizer: Consultancy; Actelion Pharmaceuticals: Research Funding.

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The train that appeared in Dutch East Indies in the mid 19th century was originally used for plantation product transport. However, the train was also used as transportation for the public. In 1894, between Batavia and Surabaya was connected with the railway line. Both cities can be reached by train in two days. To reduce travel time between those two cities, the Dutch East Indies railway company, Staatsspoorwegen was launched two high-speed trains, Eendaagsche Express in 1929 and Java Nacht Express in 1936. These two high-speed trains brought changes for railways condition in Dutch East Indies, especially in Java. During operation, these two high-speed trains also had interesting stories to write about. To write this problem, historical methods are used which consisting of heuristics, criticism, interpretation, and historiography. Based on the research conducted, these two high-speed trains made the travel time of Batavia – Surabaya 12 hours and made railways technology in the Dutch East Indies develop.

The vasculopathy of sickle cell disease (SCD), characterized by a chronic hemolytic anemia, endothelial dysfunction and recurrent vasoocclusive events, underlies most of the clinical complications. Recent work has suggested that hydroxyurea (HU) enhances endothelial nitric oxide (NO) production and reduces adhesion molecule expression which may contribute to its associated observed reduction in vasoocclusive events and improved survival in SCD. However, clinically HU is under-utilized, at least in part related to medication related toxicity, stressing a need for enhanced drug delivery. The use of nano-emulsions, a class of stable emulsions formed by a monolayer of phospholipids and/or biodegradable material such as chitosan, is a mechanism by which to enhance targeted cellular delivery of therapeutics. We hypothesized that a nano-formulation of HU would enhance drug delivery to the endothelium and may be a direct way of targeting endothelial dysfunction in SCD. Nano-formulated HU (nano-HU) was developed using a phospholipid approach and this compound was found to be biologically stable for > 6 months. Human pulmonary arterial endothelial cells (HPAECs) were co-incubated with 25µM, 125µM and 250µM HU, nano-HU, or the empty nano-emulsification preparation for 48 hours. HPAEC were isolated, and stained with propidium iodide (PI) for cell cycle analysis by FACS. HPAEC treated with 25µM nano-HU demonstrated the highest level of cell viability (55%) and similar levels of S phase inhibition as HPAEC treated with 125µM HU (where 10-20% cell viability was observed). RNA isolation of treated HPAECs was performed and qPCR for VCAM1 and SLX4, a regulator of DNA damage which promotes S phase inhibition was performed. 25µM nano-HU significantly decreased VCAM1 and increased SLX4 expression compared with 25µM HU suggesting increased efficacy at lower levels of toxicity. The impact of nano-HU on endothelial gene expression is currently being evaluated by RNA-seq. Nano-formulation of HU produces a molecule which is highly stable, and able to achieve higher intracellular concentrations with lower rates of toxicity in HPAECs .which provides the opportunity to gain greater understanding of the impact of this medication on the vascular endothelium and the potential for a more targeted and better tolerated delivery of this therapeutic to patients with sickle cell disease. Disclosures Klings: Pfizer: Consultancy.

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Tujuan penulisan artikel ini adalah untuk membangkitkan kembali memori kolektif kebencanaan gempa bumi di Jakarta. Sejarah mencatat, Jakarta yang dahulu bernama Batavia pernah beberapa kali diguncang gempa bumi. Guncangan yang paling besar yang pernah mengguncang Batavia adalah gempa bumi tahun 1699 dan 1780. Gempa bumi tahun 1699 membuat kerusakan yang cukup parah. Banyak bangunan yang mengalami kerusakan akibat gempa bumi tersebut. Kondisi ini diperparah dengan rusaknya jaringan air minum dan saluran pembuangan serta kondisi cuaca Batavia saat itu yang membuat banyak orang-orang Belanda di Batavia meninggal dunia. Banyaknya orang Belanda yang meninggal membuat Batavia saat itu mendapat julukan graf der Hollanders atau kuburan orang-orang Belanda. Bencana alam yang pernah terjadi di Batavia pada masa lalu, sangat penting untuk selalu diingat. Narasi dari sejarah bencana alam, yang dalam tulisan kali ini berfokus pada gempa bumi, dapat menjadi memori kolektif pengingat bencana. Memori kolektif ini kemudian menjadi penting sebagai salah satu cara agar masyarakat yang tinggal di daerah rawan bencana gempa bumi menjadi lebih waspada terhadap bencana yang mengintai mereka. Sebagai manusia, hendaknya sesekali melihat ke masa lalu agar dapat memahami bencana yang pernah terjadi. Dengan demikian, kita dapat mengetahui potensi bencana yang mungkin akan terjadi di masa depan dan menjadi lebih siap untuk menghadapinya.