June 20, 2014

Introduction to Biostatistics: Hypothesis Testing

Date: Thursday, August 14, 2014 Time: 1pm - 3pm EDT Duration: 120 Minutes - Online Price: $349 - Introductory Rate

UPS Honored At Annual North American 3PL Awards

UPS, an industry leader in supply chain management and the world’s largest package delivery company, was recently recognized for third-party logistics (3PL) excellence.

Insulin-regulated Glut4 Translocation [Cell Biology]

The trafficking kinetics of Glut4, the transferrin (Tf) receptor, and LRP1 were quantified in adipocytes and undifferentiated fibroblasts. Six steps were identified that determine steady state cell surface Glut4: (i) endocytosis, (ii) degradation, (iii) sorting, (iv) sequestration, (v) release, and (vi) tethering/docking/fusion. Endocytosis of Glut4 is 3 times slower than the Tf receptor in fibroblasts (ken = 0.2 min−1 versus 0.6 min−1). Differentiation decreases Glut4 ken 40% (ken = 0.12 min−1). Differentiation also decreases Glut4 degradation, increasing total and cell surface Glut4 3-fold. In fibroblasts, Glut4 is recycled from endosomes through a slow constitutive pathway (kex = 0.025–0.038 min−1), not through the fast Tf receptor pathway (kex = 0.2 min−1). The kex measured in adipocytes after insulin stimulation is similar (kex = 0.027 min−1). Differentiation decreases the rate constant for sorting into the Glut4 recycling pathway (ksort) 3-fold. In adipocytes, Glut4 is also sorted from endosomes into a second exocytic pathway through Glut4 storage vesicles (GSVs). Surprisingly, transfer from endosomes into GSVs is highly regulated; insulin increases the rate constant for sequestration (kseq) 8-fold. Release from sequestration in GSVs is rate-limiting for Glut4 exocytosis in basal adipocytes. AS160 regulates this step. Tethering/docking/fusion of GSVs to the plasma membrane is regulated through an AS160-independent process. Insulin increases the rate of release and fusion of GSVs (kfuseG) 40-fold. LRP1 cycles with the Tf receptor and Glut4 in fibroblasts but predominantly with Glut4 after differentiation. Surprisingly, AS160 knockdown accelerated LRP1 exocytosis in basal and insulin-stimulated adipocytes. These data indicate that AS160 may regulate trafficking into as well as release from GSVs.

Structures of Human ALKBH5 Demethylase Reveal a Unique Binding Mode for Specific Single-stranded N6-Methyladenosine RNA Demethylation [RNA]

N6-Methyladenosine (m6A) is the most prevalent internal RNA modification in eukaryotes. ALKBH5 belongs to the AlkB family of dioxygenases and has been shown to specifically demethylate m6A in single-stranded RNA. Here we report crystal structures of ALKBH5 in the presence of either its cofactors or the ALKBH5 inhibitor citrate. Catalytic assays demonstrate that the ALKBH5 catalytic domain can demethylate both single-stranded RNA and single-stranded DNA. We identify the TCA cycle intermediate citrate as a modest inhibitor of ALKHB5 (IC50, ∼488 μm). The structural analysis reveals that a loop region of ALKBH5 is immobilized by a disulfide bond that apparently excludes the binding of dsDNA to ALKBH5. We identify the m6A binding pocket of ALKBH5 and the key residues involved in m6A recognition using mutagenesis and ITC binding experiments.

June 19, 2014

Evolution depends on rare chance events, 'molecular time travel' experiments show

Chance events may profoundly shape history. What if Franz Ferdinand's driver had not taken a wrong turn, bringing the Duke face to face with his assassin? Would World War I still have been fought? Would Hitler have risen to power decades later?

Scientists identify link between stem cell regulation and the development of lung cancer

UCLA researchers led by Dr. Brigitte Gomperts have discovered the inner workings of the process thought to be the first stage in the development of lung cancer. Their study explains how factors that regulate the growth of adult stem cells that repair tissue in the lungs can lead to the formation of precancerous lesions.

C-to-U RNA editing

Background: RNA editing encompasses a post-transcriptional process in which the genomically templated sequence is enzymatically altered and introduces a modified base into the edited transcript. Mammalian C-to-U RNA editing represents a distinct subtype of base modification, whose prototype is intestinal apolipoproteinB mRNA, mediated by the catalytic deaminase Apobec-1. However, the genome-wide identification, tissue-specificity and functional implications of Apobec-1 mediated C-to-U RNA editing remain incompletely explored. Results: Deep sequencing, data filtering and Sanger-sequence validation of intestinal and hepatic RNA from wild-type and Apobec-1 deficient mice revealed 56 novel editing sites in 54 intestinal mRNAs and 22 novel sites in 17 liver mRNAs, all within 3[prime] untranslated regions. Eleven of 17 liver RNAs shared editing sites with intestinal RNAs, while 6 sites are unique to liver. Changes in RNA editing lead to corresponding changes in intestinal mRNA and protein levels in 11 genes. Analysis of RNA editing in vivo following tissue-specific Apobec-1 adenoviral or transgenic Apobec-1 overexpression reveals that a subset of targets identified in wild-type mice are restored in Apobec-1 deficient mouse intestine and liver following Apobec-1 rescue. We find distinctive polysome profiles for several RNA editing targets and demonstrate novel exonic editing sites in nuclear preparations from intestine but not hepatic apolipoprotein B RNA. RNA editing is validated using cell-free extracts from wild-type but not Apobec-1 deficient mice, demonstrating that Apobec-1 is required. Conclusions: These studies define selective, tissue-specific targets of Apobec-1 dependent RNA editing and show the functional consequences of editing are both transcript- and tissue-specific.

June 18, 2014

Communication With FDA: What Do We Say And How Do We Say It?

Date: Wednesday, July 23, 2014 Time: 1pm - 2:30pm EDT Duration: 90 Minutes - Online Price: $299 - Introductory Rate

FDA Approval Pushes Novartis Into 21st Century Vaccine Development

In 2009, the world experienced a global threat in the form of H1N1. Despite a prompt response to the need for a vaccine in the United States, it was still not available until six months later and not enough doses were even produced to cover all Americans. During the year-long battle with the deadly virus, the CDC estimates between 8,870 and 18,300 people died due to H1N1-related complications. In the world of influenza vaccine production using chicken eggs, the response experienced during the H1N1 outbreak is not uncommon. Is this the best our industry can do?

At the 2013 ISPE Biotechnology Symposium, Scott Billman, director of engineering at Novartis Vaccines and Diagnostics, answered this question with a resounding NO. In his presentation, he detailed the plan between Novartis and the U.S. Department of Health and Human Services to defeat influenza and any threat of future pandemics by using groundbreaking cell culture technology to produce flu vaccines. This week, the FDA granted license to the state-of-the-art Novartis Flu Cell Culture manufacturing facility in Holly Springs, NC, where the first FDA-approved vaccine using cell culture technology, Flucelvax, is being produced.

Cell culture technology uses laboratory-grown mammalian cells that are capable of hosting a growing virus. The ability to freeze these cell lines and use them later to produce a flu vaccine in large quantities is just one feature of this technology that makes it so vital in the fight against pandemics. Because cell lines grow exponentially, there is an ability to thaw them out when needed and then scale up to the manufacturing levels needed in a matter of weeks.

EMD Millipore Launches Simplicon RNA Reprogramming Technology For Efficient Generation Of Virus-Free iPSCs

EMD Millipore, the Life Science division ofMerck KGaAof Darmstadt, Germany, recently launched Simplicon RNA Reprogramming Technology, which uses synthetic self-replicating RNA to create large numbers of human induced pluripotent stem cells (iPSCs) using a single transfection step.

June 17, 2014

Bioresearch Online Editorial Board

Bioresearch Online Editorial Board

June 16, 2014

Oncobiologics Secures EU Approval For Humira Biosimilar Trial

Oncobiologics announced that it has secured approval in the EU to start a Phase I clinical trial for its Humira biosimilar molecule ONS-3010.

AstraZeneca Buys Synairgen Asthma Drug For $7.25M

AstraZeneca announced that it has signed into a global license agreement with Synairgen, a pharmaceutical company focused on respiratory disease, to buy SNG001, a novel immuno-modulatory therapy for viral-induced exacerbation in asthma.

Results of First Ever REMOTE Clinical Trial

In the clinical research industry, we’re always evaluating things. We evaluate drugs and biologics. We evaluate medical devices. We also evaluate research techniques. Yes, we try trials. We study studies. That’s what Pfizer was doing when it launched its REMOTE pilot trial in early 2011 – evaluating the effectiveness of a study in which patients participated remotely, without ever having to visit a study site. How would the results of the REMOTE study compare with those of a similar, completed, conventional study? Would this innovative research approach be validated? ByLaurie Meehan,Polaris Compliance Consultants, Inc.