Sterile media hold scale-up using MOBIUS® single-use technology

The benefits of single-use systems (SUS) in biopharmaceutical manufacturing are well understood, and their use is widespread in the manufacture of mAb, rProteins, and related therapies. New frontiers in medicine such as cell and gene therapy present an opportunity for SUS to enable speed to the clinic, however some unique hurdles must be overcome. This poster outlines the collaborative development of a single-use process to address the challenge of supplying sterile media to a bioreactor for inoculation and growth of human tissue cells. To alleviate time constraints, cleaning concerns, and contamination risks, the biopharmaceutical manufacturer chose to employ single-use technology when conducting a 5-fold scale up from a glass bottle process. A significant challenge with this human tissue cell culture process is the 60-day sterile media hold at the cell culture temperature of 36oC, during which time the bioreactor is intermittently perfused with fresh media. The Mobius® Mix50 single-use mixer (SUM) solves this challenge by first beginning with a sterile, gamma-irradiated mixer bag to eliminate concerns over validation of CIP and SIP cycles. Next, a low-pressure overlay is maintained with a carefully-sized hydrophobic vent filter, to prevent contaminants from entering the sealed mixer container. Process variables requiring assessment for this application include the air overlay pressure and flow rate, the liquid (media) flow rate during filling and draining of the SUM, sizing of the vent filter area, the liquid volume in the SUM, and the sterile condensate collection rate. A series of experiments provided a repeatable and scalable single-use solution for implementation into the manufacturing process. This novel application demonstrates the flexibility of single-use in the rapidly expanding clinical market of products derived from human cells with the unique challenges they present

GRETCHEN’S SOLILOQUY “ACH NEIGE, DU SCHMERZENREICHE” FROM GOETHE’S \u3cem\u3eFAUST\u3c/em\u3e: A VOCAL PERFORMANCE ANALYSIS AND SET OF PERFORMANCE GUIDELINES FOR VARIOUS SOLO VOICE SETTINGS

The great novelist and poet Johann Wolfgang von Goethe (1749 – 1832) arguably made his most significant contribution to the artistic world with his literary masterpiece Faust I. Goethe’s love of music and melody is evident throughout all of Faust, particularly in the expressive poetry of the character of Gretchen, whose meaningful words gave inspiration to a variety of musical manifestations, especially in German Lied. This document serves as a performance guide for vocalists. It provides vital information on the setting and arrangement of the poetry within the musical settings, the background and significance of the composer and his works, and the organization of the music. The examination of each piece will involve assessment of musical phrasing, tessitura, and overall vocal complexity in eight German Lieder settings of Gretchen’s soliloquy “Ach neige, du Schmerzenreiche” from Goethe’s Faust. The suggestions within the investigations are based on examination of pedagogical practices as well as personal experience and discoveries made while singing and performing these pieces. Through an investigation of each piece, the singer will be able to attain a successful understanding of the framework and approach to the music and poetry, and thereby achieve awareness of accurate performance practice. This document examines in order of composition, the settings by Bettina von Arnim, Franz Schubert (including the completed fragment as arranged by Benjamin Britten), Conrad Kreutzer, Bernhard Klein, Johann Loewe, Robert Schumann, Hugo Wolf, and Fredric Joseph Kroll. Because this document serves to investigate only German Lieder settings, it will not examine the choral works of Hans Pfitzner, Antoni Radziwill, Julius Röntgen, Giuseppe Verdi’s Italian setting “Deh, pietoso, oh Addolorata,” nor Richard Wagner’s Melodram. This document will also very briefly discuss the lost and inaccessible settings of Gretchen’s prayer, including those of Carl Debrois van Bruyck, Edmund von Freyhold, Moritz Hauptmann, Justus Lecerf, Leopold Lenz, Louis Schlottmann, and Hans Sommer

Marquette University Slavic Institute Papers NO. 11

https://epublications.marquette.edu/mupress-book/1011/thumbnail.jp

Graphical representation of data for a multiprocessor platform emulating spiking neural networks

Research in the eld of simulating large-scale spiking neural networks (SNN) has been carried out within the frame of Perplexus a European-funded re- search project based on a university consortium. In this project, a semi- custom electronic device called Ubichip has been designed. The mode of interest of this chip to emulate SNNs is based on a SIMD (Single-Instruction Multiple-Data) multiprocessor machine. The software for generating the as- sembly containing simulation of Iglesias-Villa spiking neural network model was also developed within that project and it is currently being successfully used for running neural network emulation on Ubichip. The tools developed so far are useful for debugging by simulation, but in order to evaluate the behavior of SNN being emulated, two needs arose: real- time monitoring of the network evolution and a higher-level, understandable visualization solution. First, the existing software that was developed in the Perplexus project has been analyzed. After examining all available solutions, including writing a standalone dedicated program, it was nally decided to develop the so-called Ubiplot plug-in. The reason was to take advantage of the existing Ubimanager environment. The development started by verifying the communication with the Ubichip, so simple waveforms for data in a given address in the Ubichip's RAM were implemented. Then the plug-in was extended with histogram and raster plots that are accessing multiple locations of the memory in each execution step. This led to the creation of the variable map that de nes the program's variables and their precise placement in the RAM. At the end simple logging facility and possibility to save and restore the layout of the plots were added. This thesis describes the Ubiplot and the development e ort put in its creation

Graphical representation of data for a multiprocessor platform emulating spiking neural networks

Research in the eld of simulating large-scale spiking neural networks (SNN) has been carried out within the frame of Perplexus a European-funded re- search project based on a university consortium. In this project, a semi- custom electronic device called Ubichip has been designed. The mode of interest of this chip to emulate SNNs is based on a SIMD (Single-Instruction Multiple-Data) multiprocessor machine. The software for generating the as- sembly containing simulation of Iglesias-Villa spiking neural network model was also developed within that project and it is currently being successfully used for running neural network emulation on Ubichip. The tools developed so far are useful for debugging by simulation, but in order to evaluate the behavior of SNN being emulated, two needs arose: real- time monitoring of the network evolution and a higher-level, understandable visualization solution. First, the existing software that was developed in the Perplexus project has been analyzed. After examining all available solutions, including writing a standalone dedicated program, it was nally decided to develop the so-called Ubiplot plug-in. The reason was to take advantage of the existing Ubimanager environment. The development started by verifying the communication with the Ubichip, so simple waveforms for data in a given address in the Ubichip's RAM were implemented. Then the plug-in was extended with histogram and raster plots that are accessing multiple locations of the memory in each execution step. This led to the creation of the variable map that de nes the program's variables and their precise placement in the RAM. At the end simple logging facility and possibility to save and restore the layout of the plots were added. This thesis describes the Ubiplot and the development e ort put in its creation

Characterization of the Electronic Excited-State Energetics and Solution Structure of Lanthanide(III) Complexes with the Polypyridine Ligand 6,6\u27-Bis[bis(2-pyridylmethyl)aminomethyl]-2,2\u27-bipyridine

Absorption, emission, and excitation spectra for solid-state and solution of Tb(III), Dy(III), and Gd(III) complexes with the polypyridine ligand 6,6‘-bis[bis(2-pyridylmethyl)-aminomethyl]-2,2‘-bipyridine (C36H34N8) are presented. Measurements of excited-state lifetimes and quantum yields in various solvents at room temperature and 77 K are also reported and used to characterize the excited-state energetics of this system. Special attention is given to the characterization of metal-to-ligand energy transfer efficiency and mechanisms. The measurement of circularly polarized luminescence (CPL) from the solution of the Dy(III) complex following circularly polarized excitation confirms the chiral structure of the complexes under study. No CPL is present in the luminescence from the Eu(III) or Tb(III) complex because of efficient racemization. The variation of the magnitude of the CPL as a function of temperature from an aqueous solution of DyL is used for the first time to characterize the solution equilibria between different chiral species

Assessing environmentally effective post‑COVID green recovery plans for reducing social and economic inequality

Given the current environmental crisis there have been multiple calls for a green recovery from COVID-19 which address environmental concerns and provide jobs in industries and communities economically damaged by the pandemic. Here, we holistically evaluate a range of recovery scenarios, evaluated on environmental and socio-economic equity metrics. Using a modified version of a Bayesian belief network, we show that economic stimuli across green sectors, including jobs in renewable energy, waste management, retrofitting of buildings, heat-pump installation and public transport can help economic growth, but will have limited environmental benefits. The inclusion of carbon taxes and ending fossil fuel subsidies, alongside investment in nature-based solutions and jobs in ecological conservation, can greatly increase the environmental gains as well as socio-economic equality. Additionally, jobs not associated with green industries, but with low carbon footprints, such as those in social care can further improve social equality with minimal negative environmental effects. However, in these latter scenarios involving taxation and ending fossil fuel subsidies, economic growth is reduced. We suggest a comprehensive green recovery and green new deal are needed, and we should reimagine economies, without the focus on economic growth

Multiplex Cytological Profiling Assay to Measure Diverse Cellular States

Computational methods for image-based profiling are under active development, but their success hinges on assays that can capture a wide range of phenotypes. We have developed a multiplex cytological profiling assay that “paints the cell” with as many fluorescent markers as possible without compromising our ability to extract rich, quantitative profiles in high throughput. The assay detects seven major cellular components. In a pilot screen of bioactive compounds, the assay detected a range of cellular phenotypes and it clustered compounds with similar annotated protein targets or chemical structure based on cytological profiles. The results demonstrate that the assay captures subtle patterns in the combination of morphological labels, thereby detecting the effects of chemical compounds even though their targets are not stained directly. This image-based assay provides an unbiased approach to characterize compound- and disease-associated cell states to support future probe discovery

Reactivity of a series of triaryl borates, B(OAr x) 3, in hydroboration catalysis

In this paper, we compare the reactivity of a series of triaryl borates B(OArx)3 as catalysts for the hydroboration of alkenes and alkynes. It was observed that commercially available B(OPh)3 performed the poorest, whereas catalysts with o-F atoms appeared to perform much better

A novel small-caliber bacterial cellulose vascular prosthesis: production, characterization, and preliminary in vivo testing

Vascular grafts are used to bypass damaged or diseased blood vessels. Bacterial cellulose (BC) has been studied for use as an off-the-shelf graft. Herein, we present a novel, cost-effective, method for the production of small caliber BC grafts with minimal processing or requirements. The morphology of the graft wall produced a tensile strength above that of native vessels, performing similarly to the current commercial alternatives. As a result of the production method, the luminal surface of the graft presents similar topography to that of native vessels. We have also studied the in vivo behavior of these BC graft in order to further demonstrate their viability. In these preliminary studies, 1 month patency was achieved, with the presence of neo-vessels and endothelial cells on the luminal surface of the graft.This study was supported by the Portuguese Foundation for Science and Technology (FCT) and the European Community fund FEDER, through Program COMPETE, under the scope of the Projects FCOMP-01-0124-FEDER-007025 (PTDC/AMB/68393/2006), PEST-OE/EQB/LA0023/2013, PEST-C/FIS/UI607/2013, RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and the Projects "BioEnv-Biotechnology and Bioengineering for a sustainable world" and "Matepro-Optimizing Materials and Processes". NORTE-07-0124-FEDER-000048, co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER. The authors also acknowledge the fellowship awarded to Alexandre Felipe Leitao (SFRH/BD/66094/2009) funded by the Fundacao para a Ciencia e Tecnologia (FCT). The authors also thank support by FCT through the project BCGrafts, FCOMP-01-0124-FEDER-014773 (PTDC/EBB/EBI/112170/2009) and by the People Program (Marie Curie Actions) of the European Union's Seventh Framework Program FP7/2007-2013/under REA grant agreement n317512