Rebels with a cause. Aldo van Eyck and Pancho Guedes, how to find a meaning for the act of built

This paper aims to look at an uncommonly critical attitude against the bureaucratic functionalism in force within a kind of International Style, developing an authentically modern and human architecture in the scope of the Team 10’s battle. Considering Aldo van Eyck (1918-1999) and Pancho Guedes’ (1925-2015) works and thoughts, their parallel paths, sometimes crossed, are analysed: they were both part of Team 10 and they both defined architecture as the “built meaning”, recalling its multiple meanings, languages and responsibilities: ‘I claim for architects the rights and liberties that painters and poets have held for so long’. Aldo van Eyck, from the studies on the sub-Saharan Dogon region to the PREVI proposals in Peru, and Pancho Guedes, from the survey on the Mapogga doors to his surrealist approaches in Mozambique, give examples of the transformation process, on how the modern project got elasticity, creativity, endurance, and finally feeding the utopia. The argument addresses the fact that these two minds envisaged architecture as a language with an emotional impact and a social and cultural scope. Bearing in mind architecture as the primary visual medium with which human society expresses and reveals itself, architecture is conceived as a dialogue and the design of buildings as means for creating relations between people rather than as an end in itself.info:eu-repo/semantics/publishedVersio

La permanente experimentación en Álvaro Siza

[ES] Álvaro Siza (1933) es el arquitecto portugués más importante de su generación. Así lo afi rmaba Peter Testa en 1984, en la introducción de su libro dedicado a la obra de Álvaro Siza. Hoy, pasados ya casi 25 años, podemos afi rmar sin miedo a exagerar que Álvaro Siza es sencillamente uno de los arquitectos más importantes de nuestra contemporaneidad. Con trabajos realizados desde Corea a Santiago de Compostela, desde Porto Alegre a La Haya o Berlín, Siza ha construido un recorrido único, aliando una actitud de permanente experimentación sobre los sitios, los programas o los modos de construir, a una extraordinaria capacidad lírica.Tostoes, A. (2008). La permanente experimentación en Álvaro Siza. EN BLANCO. Revista de Arquitectura. 1(1):6-11. doi:10.4995/eb.2008.7284SWORD6111

REUSE OF COMMON SPACE AS A TACTIC FOR MASS HOUSING REVITALIZATION

Urban decay and obsolescence of post-war mass housing is a global phenomenon. Although the reasons for housing deterioration are different, the altered relationship between public and private spaces is essential for the mass housing. The research hypothesizes that strong polarisation of the urban landscape into private and public is firmly influencing urban decay and obsolescence of post-war mass housing neighbourhoods. Taking New Belgrade blocks as the case study, the research investigates this correlation, following the gradual transformation of the urban landscape of modernity in parallel with different factors. Moreover, the research sheds light on the specific Yugoslav housing policy and developed collective self-management of the urban commons from the time of construction. Although these strategies have been neglected over the time, they are valuable for contemporary, increasing discussions on community-driven approaches for comprehending and managing change in urban environment, specifically for residential neighbourhoods. Furthermore, the research is analysing different contemporary strategies and community practices that are reinventing the public-private relationship in the context of mass housing, contributing to the development of a methodology for mitigating obsolescence and causes of housing deterioration. The methodology is revitalizing the important value of common spaces and the role of community and is reusing the modernist idea of co-creation, contributing to inheritance of the modernist concepts. Moreover, if applied, it would increase liveability of urban space and well-being of its residents, contributing to transformation strategies for adaptation to current needs, and therefore ensuring vitality of mass housing as a core typology of the Modern Movement

A novel filtration system for point of care washing of cellular therapy products

The cell therapy industry would greatly benefit from a simple point of care solution to remove Dimethyl Sulfoxide (DMSO) from small volume thawed cell suspensions prior to injection. We have designed and validated a novel dead-end filtration device, which takes advantage of the higher density of thawed cell suspensions to remove the DMSO and protein impurities from the cell suspension without fouling the filter membrane. The filter was designed to avoid fluid circuits and minimize the surface area that is contacted by the cell suspension, thus reducing cell losses by design. The filtration process was established through optimization of the fluid flow configuration, backflush cycles and filter geometry. Overall, this novel filtration device allows for a 1 mL of thawed cryopreserved cell suspensions, containing 107 cells of a foetal lung fibroblast cell line (MRC-5), to be washed in less than 30 minutes. More than 95% of the DMSO and up to 94% of the Albumin- Fluorescein-Isothiocyanate content can be removed while the viable cell recovery is higher than 80%. We have also demonstrated that this system can be used for bone marrow-derived human mesenchymal stem cells with more than 73% cell recovery and 85% DMSO reduction. This is the first time that a dead end (normal) filtration process has been used to successfully wash high density human cell suspensions. In practice, this novel solid-liquid separation technology fills the need for small volume washing in closed processing systems for cellular therapies

Early retinal differentiation of human pluripotent stem cells in microwell suspension cultures

OBJECTIVE: To develop a microwell suspension platform for the adaption of attached stem cell differentiation protocols into mixed suspension culture. RESULTS: We adapted an adherent protocol for the retinal differentiation of human induced pluripotent stem cells (hiPSCs) using a two-step protocol. Establishing the optimum embryoid body (EB) starting size and shaking speed resulted in the translation of the original adherent process into suspension culture. Embryoid bodies expanded in size as the culture progressed resulting in the expression of characteristic markers of early (Rx, Six and Otx2) and late (Crx, Nrl and Rhodopsin) retinal differentiation. The new process also eliminated the use of matrigel, an animal-derived extracellular matrix coating. CONCLUSIONS: Shaking microwells offer a fast and cost-effective method for proof-of-concept studies to establish whether pluripotent stem cell differentiation processes can be translated into mixed suspension culture

Volume reduction, cell washing and affinity cell selection using multi-dimensional acoustic standing wave technology

Acoustic Cell Processing is a unique acousto-fluidics platform technology for shear-free manipulation of cells using ultrasonic standing waves. The platform has broad applications in the field of cell and gene therapy, e.g., cell concentration and washing, cell culturing, microcarrier/cell separation, acoustic affinity cell selection and label-free cell selection. The acoustic radiation force exerted by the ultrasonic standing wave on the suspended cells in combination with fluid drag forces and gravitational forces is used to manipulate the cells and achieve a certain cell processing unit operation, e.g., separate, concentrate, or wash. The technology is single-use, continuous, and can be scaled up, down or out. It therefore allows for a flexible and modular approach that can be customized to process a desired cell count, cell culture volume or cell concentration within a given required process time. Utilizing its proprietary multi-dimensional standing wave platform, FloDesign Sonics (FD Sonics) has been developing two applications for cell and gene therapy manufacturing, an Acoustic Concentrate-Wash (ACW) and Acoustic Affinity Cell Selection (AACS) system for closed and shear free Cell and Gene Therapy manufacturing, namely CAR-T immunocellular therapies. The ACW technology has been applied to Jurkat T-cells and primary cultures of T-cells of 1-2 Liters (L) with cell concentrations ranging from 1 million cells per milliliter (ml) to 40 million cells per ml. The process flow rate varies from 2-3 L/hour with average cell recoveries of more than 80% in 60 to 90 minutes. The efficiency of the cell washing process ranges from 95-99% depletion of a model protein (BSA), depending on the wash methodology. The AACS technology is a scalable acoustic affinity cell selection method using acoustic (non-paramagnetic) affinity beads for positive or negative cell selection. A multi-dimensional acoustic standing wave is then used to separate the affinity bead-cell complexes from the unbound cells, thereby completing the process of a negative or positive cell selection. A population of 1 billion CAR-T cells containing 30% T-Cell Receptor positive (TCR+) and 70% T-cell Receptor Negative (TCR-) cells has been depleted of 99% of its TCR+ population. The TCR- cell recovery for this process was above 70% and the full process took less than 2 hours. When used for positive selection of CD3+ cells, AACS allowed for an enrichment of 2.5-fold in CD3+ population. ACW and AACS are powerful acoustic-based cell processing technologies that lower cost and risk while enabling a modular, automation-friendly manufacturing process for cell and gene therapy manufacturing. Please click Additional Files below to see the full abstract

Oxygen-controlled automated neural differentiation of mouse embryonic stem cells.

Automation and oxygen tension control are two tools that provide significant improvements to the reproducibility and efficiency of stem cell production processes. Aim: the aim of this study was to establish a novel automation platform capable of controlling oxygen tension during both the cell-culture and liquid-handling steps of neural differentiation processes. Materials & methods: We built a bespoke automation platform, which enclosed a liquid-handling platform in a sterile, oxygen-controlled environment. An airtight connection was used to transfer cell culture plates to and from an automated oxygen-controlled incubator. Results: Our results demonstrate that our system yielded comparable cell numbers, viabilities, metabolism profiles and differentiation efficiencies when compared with traditional manual processes. Interestingly, eliminating exposure to ambient conditions during the liquid-handling stage resulted in significant improvements in the yield of MAP2-positive neural cells, indicating that this level of control can improve differentiation processes. Conclusion: This article describes, for the first time, an automation platform capable of maintaining oxygen tension control during both the cell-culture and liquid-handling stages of a 2D embryonic stem cell differentiation process

Long‐Term Retinal Differentiation of Human Induced Pluripotent Stem Cells in a Continuously Perfused Microfluidic Culture Device

Understanding how microenvironmental cues influence cellular behavior will enable development of efficient and robust pluripotent stem cell differentiation protocols. Unlike traditional cell culture dishes, microfluidic bioreactors can provide stable microenvironmental conditions by continuous medium perfusion at a controlled rate. The aim of this study is to investigate whether a microfluidic culture device could be used as a perfused platform for long‐term cell culture processes such as the retinal differentiation of human induced pluripotent stem cells. The perfusion flow rate is established based on the degradation and consumption of growth factors (DKK‐1, Noggin, IGF‐1, and bFGF) and utilizing the Péclet number. The device's performance analyzed by qRT‐PCR show improvements compared to the well‐plate control as characterized by significantly higher expression of the markers Pax6, Chx10, and Crx on Day 5, Nrl on day 10, Crx, and Rhodopsin on day 21. Optimization of perfusion rate is an important operating variable in development of robust processes for differentiation cultures. Result demonstrates convective delivery of nutrients via perfusion has a significant impact upon the expression of key retinal markers. This study is the first continuously perfused long‐term (21 days) retinal differentiation of hiPSCs in a microfluidic device

Entrevista a Álvaro Siza

[ES] En sus primeras obras, el restaurante de Boa Nova y las piscinas de Leça de Palmeira, utilizó usted el hormigón visto, y tras un largo periodo de abandono de este material, lo ha retomado en sus últimos edificios adoptando ahora el hormigón blanco, ¿Cual es su relación con ese material?Mas Llorens, V.; García-Gasco Lominchar, S.; Silvestre Navarro, F.; Tostoes, A. (2008). Entrevista a Álvaro Siza. EN BLANCO. Revista de Arquitectura. 1(1):12-17. doi:10.4995/eb.2008.7285SWORD12171

Characterization of a functional C3A liver spheroid model

More predictive in vitro liver models are a critical requirement for preclinical screening of compounds demonstrating hepatotoxic liability. 3D liver spheroids have been shown to have an enhanced functional lifespan compared to 2D monocultures; however a detailed characterisation of spatiotemporal function and structure of spheroids still needs further attention before widespread use in industry. We have developed and characterized the structure and function of a 3D liver spheroid model formed from C3A hepatoma cells. Spheroids were viable and maintained a compact in vivo-like structure with zonation features for up to 32 days. MRP2 and Pgp transporters had polarised expression on the canalicular membrane of cells in the spheroids and were able to functionally transport CMFDA substrate into these canalicular structures. Spheroids expressed CYP2E1 and were able to synthesise and secrete albumin and urea to a higher degree than monolayer C3A cultures. Penetration of doxorubicin throughout the spheroid core was demonstrated. Spheroids showed increased susceptibility to hepatotoxins when compared to 2D cultures, with acetaminophen having an IC50 of 7.2 mM in spheroids compared to 33.8 mM in monolayer culture. To conclude, we developed an alternative method for creating C3A liver spheroids and demonstrated cellular polarisation and zonation, as well as superior liver-specific functionality and more sensitive toxicological response compared to standard 2D liver models, confirming a more in vivo-like liver model