1,982 research outputs found
Risk Doctor Briefing - The definition debate continued
Risk Doctor Briefing - The definition debate continue
Cell culture engineering and biosimilars: The physician’s perspective
Biosimilars promise to increase access to life-saving and life-enhancing therapeutics, while creating cost savings that can fund further innovations in health care. Achievement of these goals is critically dependent on acceptance by prescribing physicians. This session will explore the following questions: What do clinicians care about? What do they know about the manufacture and characterization of biosimilars? What should they know? These questions will be addressed in the context of specific examples arising in the development of biosimilars to Neupogen, Infliximab, Humira, and Rituximab
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Fluorescent amplification for next generation sequencing (FA-NGS) library preparation.
BACKGROUND:Next generation sequencing (NGS) has become a universal practice in modern molecular biology. As the throughput of sequencing experiments increases, the preparation of conventional multiplexed libraries becomes more labor intensive. Conventional library preparation typically requires quality control (QC) testing for individual libraries such as amplification success evaluation and quantification, none of which occur until the end of the library preparation process. RESULTS:In this study, we address the need for a more streamlined high-throughput NGS workflow by tethering real-time quantitative PCR (qPCR) to conventional workflows to save time and implement single tube and single reagent QC. We modified two distinct library preparation workflows by replacing PCR and quantification with qPCR using SYBR Green I. qPCR enabled individual library quantification for pooling in a single tube without the need for additional reagents. Additionally, a melting curve analysis was implemented as an intermediate QC test to confirm successful amplification. Sequencing analysis showed comparable percent reads for each indexed library, demonstrating that pooling calculations based on qPCR allow for an even representation of sequencing reads. To aid the modified workflow, a software toolkit was developed and used to generate pooling instructions and analyze qPCR and melting curve data. CONCLUSIONS:We successfully applied fluorescent amplification for next generation sequencing (FA-NGS) library preparation to both plasmids and bacterial genomes. As a result of using qPCR for quantification and proceeding directly to library pooling, the modified library preparation workflow has fewer overall steps. Therefore, we speculate that the FA-NGS workflow has less risk of user error. The melting curve analysis provides the necessary QC test to identify and troubleshoot library failures prior to sequencing. While this study demonstrates the value of FA-NGS for plasmid or gDNA libraries, we speculate that its versatility could lead to successful application across other library types
The Standard of Proof in Juvenile Proceedings: \u3cem\u3eGault\u3c/em\u3e Beyond a Reasonable Doubt
Some of those who have studied the question of the appropriate standard of proof in juvenile proceedings have determined that the preponderance of the evidence standard-the standard applied in civil cases-is sufficient, and that the criminal standard should not be applied in such cases. Others have suggested that the standard-of proof question is unimportant since the particular standard which is required will seldom, if ever, make a difference to the outcome of a case. The first of these views is the subject to which the bulk of this Article is addressed; the second can be rebutted by the observation that in at least two recent cases youths were found to be delinquent by judges who specifically stated that their conclusions would have been different if the higher standard had been applicable
(The) Jewish motives in Heine's works ..
Typewritten sheets in cover.
Thesis (M.A.)--Boston University
Bibliography: 3 p. at end
An integrated machine learning and experimental approach to uncover ageing-associated processes in Fission Yeast
This work attempts to bring together knowledge of different pathways associated with cellular ageing and create connections between them using both machine learning and experimental methods. Initially, I describe the development of a novel proxy for chronological lifespan as part of the analysis pipeline of a high-throughput chronological lifespan assay in fission yeast. I then use this technique to go on to develop novel machine learning models that can predict lifespan, a complex phenotype, from simple traits, and identify ageing-associated phenotypes in fission yeast.
Complementary to this, I investigate a transcription factor of interest, Hsr1, for its involvement in cellular ageing and ageing-associated processes. I describe direct regulatory targets and how it forms a network with at least four other ageing-associated transcription factors which bridges the gaps between models of ageing, and suggest mechanisms for these interactions.
In this way, this work provides novel links between cellular ageing mechanisms and ageing-associated processes from both machine learning and experimental sources
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Enhancing Terminal Deoxynucleotidyl Transferase Activity on Substrates with 3' Terminal Structures for Enzymatic De Novo DNA Synthesis.
Enzymatic oligonucleotide synthesis methods based on the template-independent polymerase terminal deoxynucleotidyl transferase (TdT) promise to enable the de novo synthesis of long oligonucleotides under mild, aqueous conditions. Intermediates with a 3' terminal structure (hairpins) will inevitably arise during synthesis, but TdT has poor activity on these structured substrates, limiting its usefulness for oligonucleotide synthesis. Here, we described two parallel efforts to improve the activity of TdT on hairpins: (1) optimization of the concentrations of the divalent cation cofactors and (2) engineering TdT for enhanced thermostability, enabling reactions at elevated temperatures. By combining both of these improvements, we obtained a ~10-fold increase in the elongation rate of a guanine-cytosine hairpin
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