255 research outputs found

    Code and its image: the functions of text and visualisation in a code-based design studio

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    Traditionally, design learning in the architecture studio has taken place through a combination of individual work and joint projects. The introduction of code-based design practices in the design studio has altered this balance, introducing new models of joint authorship and new ways for individuals to contribute to co-authored projects. This paper presents a case study describing four design studios in a higher education setting that used code as a tool for generating architectural geometry. The format of the studios encouraged the students to reflect critically on their role as authors and to creatively address the multiple opportunities for shared authorship available with code-based production. The research question addressed in this study involved the role of code-based practices in altering the model of architectural education in the design studio, in particular the role of visual representations of a code-based design process in the production of shared knowledge

    Modeling Pichia pastoris Growth on Methanol and Optimizing the Production of a Recombinant Protein, the Heavy-Chain Fragment C of Botulinum Neurotoxin, Serotype A

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    An unstructured growth model for the recombinant methylotrophic yeast P. pastoris Mut+ expressing the heavy-chain fragment C of botulinum neurotoxin serotype A [BoNT/A(Hc)], was successfully established in quasi-steady state fed-batch fermentations with varying cell densities. The model describes the relationships between specific growth rate and methanol concentration, and the relationships between specific methanol and ammonium consump-tion rates and specific growth rate under methanol-limited growth conditions. The maximum specific growth rate (μ) determined from the model was 0.08 h−1 at a methanol concentration of 3.65 g/L, while the actual maximum μ was 0.0709 h−1. The maximum specific methanol consumption rate was 0.0682 g/g WCW/h. From the model, growth can be defined as either methanol-limited or metha-nol-inhibited and is delineated at a methanol concentration of 3.65 g/L. Under inhibited conditions, the observed biomass yield (YX/MeOH) was lower and the maintenance coefficient (mMeOH) was higher than compared to limited methanol conditions. The YX/MeOH decreased and mMeOH increased with increasing methanol concen-tration under methanolinhibited conditions. BoNT/A(Hc) content in cells (a) under inhibited growth was lower than that under limited growth, and decreased with increasing methanol concentration. A maximum a of 1.72 mg/g WCW was achieved at a μ of 0.0267 h−1 and induction time of 12 h

    Crystallization and preliminary X-ray analysis of \u3ci\u3eNa\u3c/i\u3e-ASP-1, a multi-domain pathogenesis-related-1 protein from the human hookworm parasite \u3ci\u3eNecator americanus\u3c/i\u3e

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    Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis-related-1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been expressed, purified and crystallized. Na-ASP-1 is a 406-amino-acid multi-domain ASP from the prevalent human hookworm parasite Necator americanus. Useful X-ray data to 2.2 A ° have been collected from a crystal that belongs to the monoclinic space group P21 with unit-cell parameters a = 67.7, b = 74.27, c = 84.60 Å, β = 112.12°. An initial molecular-replacement solution has been obtained with one monomer in the asymmetric unit

    Crystallization and preliminary X-ray analysis of \u3ci\u3eNa\u3c/i\u3e-ASP-1, a multi-domain pathogenesis-related-1 protein from the human hookworm parasite \u3ci\u3eNecator americanus\u3c/i\u3e

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    Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis-related-1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been expressed, purified and crystallized. Na-ASP-1 is a 406-amino-acid multi-domain ASP from the prevalent human hookworm parasite Necator americanus. Useful X-ray data to 2.2 A ° have been collected from a crystal that belongs to the monoclinic space group P21 with unit-cell parameters a = 67.7, b = 74.27, c = 84.60 Å, β = 112.12°. An initial molecular-replacement solution has been obtained with one monomer in the asymmetric unit

    Cell bank characterization and fermentation optimization for production of recombinant heavy chain C-terminal fragment of botulinum neurotoxin serotype E (rBoNTE(H\u3csub\u3ec\u3c/sub\u3e): Antigen E) by \u3ci\u3ePichia pastoris\u3c/i\u3e

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    A process was developed for production of a candidate vaccine antigen, recombinant C-terminal heavy chain fragment of the botulinum neurotoxin serotype E, rBoNTE(Hc)in Pichia pastoris. P. pastoris strain GS115 was transformed with the rBoNTE(Hc) gene inserted into pHILD4 Escherichia coli—P. pastoris shuttle plasmid. The clone was characterized for genetic stability, copy number, and BoNTE(Hc) sequence. Expression of rBoNTE(Hc) from the Mut+ HIS4 clone was confirmed in the shake-flask, prior to developing a fed-batch fermentation process at 5 and 19 L scale. The fermentation process consists of a glycerol growth phase in batch and fed-batch mode using a defined medium followed by a glycerol/methanol transition phase for adaptation to growth on methanol and a methanol induction phase resulting in the production of rBoNTE(Hc). Specific growth rate, ratio of growth to induction phase, and time of induction were critical for optimal rBoNTE(Hc) production and minimal proteolytic degradation. A computer-controlled exponential growth model was used for process automation and off-gas analysis was used for process monitoring. The optimized process had an induction time of 9 h on methanol and produced up to 3 mg of rBoNTE(Hc) per gram wet cell mass as determined by HPLC and Western blot analysis

    Cell bank characterization and fermentation optimization for production of recombinant heavy chain C-terminal fragment of botulinum neurotoxin serotype E (rBoNTE(H\u3csub\u3ec\u3c/sub\u3e): Antigen E) by \u3ci\u3ePichia pastoris\u3c/i\u3e

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    A process was developed for production of a candidate vaccine antigen, recombinant C-terminal heavy chain fragment of the botulinum neurotoxin serotype E, rBoNTE(Hc)in Pichia pastoris. P. pastoris strain GS115 was transformed with the rBoNTE(Hc) gene inserted into pHILD4 Escherichia coli—P. pastoris shuttle plasmid. The clone was characterized for genetic stability, copy number, and BoNTE(Hc) sequence. Expression of rBoNTE(Hc) from the Mut+ HIS4 clone was confirmed in the shake-flask, prior to developing a fed-batch fermentation process at 5 and 19 L scale. The fermentation process consists of a glycerol growth phase in batch and fed-batch mode using a defined medium followed by a glycerol/methanol transition phase for adaptation to growth on methanol and a methanol induction phase resulting in the production of rBoNTE(Hc). Specific growth rate, ratio of growth to induction phase, and time of induction were critical for optimal rBoNTE(Hc) production and minimal proteolytic degradation. A computer-controlled exponential growth model was used for process automation and off-gas analysis was used for process monitoring. The optimized process had an induction time of 9 h on methanol and produced up to 3 mg of rBoNTE(Hc) per gram wet cell mass as determined by HPLC and Western blot analysis

    cGMP Recombinant FIX for IV and Oral Hemophilia B Therapy

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    Three specific aims are proposed: Specific Aim # 1. Process engineer and scale-up the recovery and purification of transgenic recombinant human Factor IX. The University of Nebraska-Lincoln Biological Process Development Facility will complete process development and scale-up, and produce clinical grade materials for preclinical studies. The endpoint is a proposed final product specification to help facilitate transfer to current Good Manufacturing Practices compliant production of clinical grade material to support an Investigational New Drug filing with the United States Food and Drug Administration (FDA) leading to clinical trials. Specific Aim #2. Characterize and formulate transgenic recombinant human Factor IX for intravenous dosage, and evaluate in a hemophilia B dog model. These activities are directed toward characterization of the product important to assure the provision of safe and reproducibly effective hemostasis. The results of these investigations will help support an IND filing with the FDA. Specific Aim # 3. Develop an oral dosage form of transgenic recombinant human Factor IX, and evaluate in hemophilia B mice and dog models. Oral administration of coagulation therapy will obviate the invasiveness, discomfort, potential for opportunistic infection, and complications of storage and supplies that accompany intravenous administration. Oral dosage forms of Factor IX will thus greatly increase the proportion of the patient population that can be treated. There is also published evidence suggesting that oral administration may reduce the potential for complicating immune responses to replacement therapy, especially in patients with severe hemophilia

    Mineral chemistry of igneous melanite garnets from analcite-bearing volcanic rocks, Alberta, Canada

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    The mineral chemistry of melanite garnets from the Crowsnest volcanic rocks of SW Alberta, Canada, has been investigated by using electron microprobe scans, quantitative analyses and multivariate statistical analysis. The garnets occur with aegirine-augite, sanidine, analcite and rare plagioclase as phenocrysts in trachyte and phonolite flows, agglomerates and tuffs. Wavelength dispersive microprobe scans reveal complex zonation patterns, both normal and oscillatory. The results of fifty quantitative analyses were subjected to R-mode factor analysis to delineate the chemical exchanges producing the zonation. The chemical zonation of the garnets may be attributed to four independent binary exchanges; Al-Fe3+, Si-Ti, Ca-Mn and Mg-Fe2+. The stoichiometry of these garnets, based on microprobe and wet chemical Fe analyses, combined with the strongly antithetic behavior of Si and Ti lead us to infer that the Ti in these garnets is dominantly tetravalent. It is clear from this study that quantitative modelling of the processes of crystal growth and zonation of melanite garnets in alkaline, undersaturated igneous rocks should be aimed at simulating the four chemical exchanges listed above
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