Generation and Characterization of a Tissue-Specific Centrosome Indicator Mouse Line.

Centrosomes are major microtubule organizing centers (MTOCs) that play an important role in chromosome segregation during cell division. Centrosomes provide a stable anchor for microtubules, constituting the centers of the spindle poles in mitotic cells, and determining the orientation of cell division. However, visualization of centrosomes is challenging because of their small size. Especially in mouse tissues, it has been extremely challenging to observe centrosomes belonging to a specific cell type of interest among multiple comingled cell types. To overcome this obstacle, we generated a tissue-specific centrosome indicator. In this mouse line, a construct containing a floxed neomyocin resistance gene with a triplicate polyA sequence followed by an EGFP-Centrin1 fusion cassette was knocked into the Rosa locus. Upon Cre-mediated excision, EGFP-Centrin1 was expressed under the control of the Rosa locus. Experiments utilizing mouse embryo fibroblasts (MEFs) demonstrated the feasibility of real-time imaging, and showed that EGFP-Centrin1 expression mirrored the endogenous centrosome cycle, undergoing precisely one round of duplication through the cell cycle. Moreover, experiments using embryo and adult mouse tissues demonstrated that EGFP-Centrin1 specifically mirrors the localization of endogenous centrosomes. genesis 54:286-296, 2016. © 2016 The Authors. Genesis Published by Wiley Periodicals, Inc

Fast Reachable Set Approximations via State Decoupling Disturbances

With the recent surge of interest in using robotics and automation for civil purposes, providing safety and performance guarantees has become extremely important. In the past, differential games have been successfully used for the analysis of safety-critical systems. In particular, the Hamilton-Jacobi (HJ) formulation of differential games provides a flexible way to compute the reachable set, which can characterize the set of states which lead to either desirable or undesirable configurations, depending on the application. While HJ reachability is applicable to many small practical systems, the curse of dimensionality prevents the direct application of HJ reachability to many larger systems. To address computation complexity issues, various efficient computation methods in the literature have been developed for approximating or exactly computing the solution to HJ partial differential equations, but only when the system dynamics are of specific forms. In this paper, we propose a flexible method to trade off optimality with computation complexity in HJ reachability analysis. To achieve this, we propose to simplify system dynamics by treating state variables as disturbances. We prove that the resulting approximation is conservative in the desired direction, and demonstrate our method using a four-dimensional plane model.Comment: in Proceedings of the IEE Conference on Decision and Control, 201

Business Analysis of the Women’s Professional Basketball League

http://deepblue.lib.umich.edu/bitstream/2027.42/100221/1/MBAChen1996Final.pd

Computational Identification of Four Spliceosomal snRNAs from the Deep-Branching Eukaryote Giardia intestinalis

Funding: Marsden Fund New Zealand Allan Wilson Centre The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.RNAs processing other RNAs is very general in eukaryotes, but is not clear to what extent it is ancestral to eukaryotes. Here we focus on pre-mRNA splicing, one of the most important RNA-processing mechanisms in eukaryotes. In most eukaryotes splicing is predominantly catalysed by the major spliceosome complex, which consists of five uridine-rich small nuclear RNAs (U-snRNAs) and over 200 proteins in humans. Three major spliceosomal introns have been found experimentally in Giardia; one Giardia U-snRNA (U5) and a number of spliceosomal proteins have also been identified. However, because of the low sequence similarity between the Giardia ncRNAs and those of other eukaryotes, the other U-snRNAs of Giardia had not been found. Using two computational methods, candidates for Giardia U1, U2, U4 and U6 snRNAs were identified in this study and shown by RT-PCR to be expressed. We found that identifying a U2 candidate helped identify U6 and U4 based on interactions between them. Secondary structural modelling of the Giardia U-snRNA candidates revealed typical features of eukaryotic U-snRNAs. We demonstrate a successful approach to combine computational and experimental methods to identify expected ncRNAs in a highly divergent protist genome. Our findings reinforce the conclusion that spliceosomal small-nuclear RNAs existed in the last common ancestor of eukaryotes

Directed evolution, rational design and mechanistic studies of a phosphohydrolase from Enterobacter aerogenes

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Pentingnya Akuntansi Keuangan Bagi Organisasi Nirlaba (Studi Kasus pada Kubca Samakta)

Sociopreneur dipercaya mampu membawa Perubahan ke arah yang lebih baik bagi masyarakat khususnya di Indonesia. Dengan misi sosial memberdayakan masyarakat, sociopreneur dapat menyediakan lapangan kerja bagi masyarakat yang pengangguran dan masyarakat bawah sehingga jumlah masyarakat miskin di Indonesia akan berkurang, dan kemandirian masyarakat semakin meningkat.Kubca Samakta adalah salah satu contoh lembaga nirlaba yang sangat kental dengan semangat sociopreneur. Kubca Samakta melakukan pemberdayaan terhadap para remaja/dewasa penyandang cacat khususnya tuna rungu, melalui program pelatihan keterampilan dan program unit produksi bagi para penyandang cacat. Kubca Samakta diharapkan dapat meningkatkan kemandirian para penyandang cacat untuk meraih kualitas hidup yang lebih baik.Sebagai lembaga nirlaba, Kubca Samakta dituntut untuk melakukan pertanggungjawaban yang memadai kepada para stakeholder, khususnya penyandang dana dan pemberi bantuan sumber daya lainnya. Salah satu bentuk laporan pertanggungjawaban adalah laporan keuangan yang disusun secara berkala dan merupakan output dari akuntansi keuangan. Penelitian ini dilaksanakan untuk membantu Kubca Samakta dan organisasi nirlaba sejenis dalam menerapkan akuntansi keuangan organisasi nirlaba sesuai dengan prinsip akuntansi yang berlaku umum di Indonesia.Kata Kunci : Akuntansi Organisasi Nirlaba, Sociopreneu

Molecular analysis of cDNA clones and the corresponding genomic coding sequences of the Drosophila dunce' gene, the structural gene for cAMP phosphodiesterase

We have isolated and sequenced cDNA clones representing portions of the polyadenylylated transcripts of the dunce+ gene. These define an open reading frame of 1086 bases and some of the 5'- and 3'-untranslated regions of the transcripts. The deduced amino acid sequence is strikingly homologous to the amino acid sequence of a Ca^2+/calmodulin-dependent cyclic nucleotide phosphodiesterase isolated from bovine brain and more weakly related to the predicted amino acid sequence of a yeast cAMP phosphodiesterase. These homologies, together with prior genetic and biochemical studies, provide unambiguous evidence that dunce^+ codes for a phosphodiesterase. In addition, the dunce^+ gene product shares a seven-amino acid sequence with a regulatory subunit of cAMP-dependent protein kinase that is predicted to be part of the cAMP binding site. We also identify a weak homology between a region of the dunce+ gene product and the egg-laying hormone precursor of Aplysia californica. The open reading frame is divided in the genome by four introns