Hasta Que Haya Justicia: Responses to the 2014 Forced Disappearance of the Students of Ayotzinapa, Mexico

This paper examines responses to the 2014 forced disappearance of 43 university students from Ayotzinapa, Guerrero by Mexican authorities and criminals working together, for reasons that remain unknown. Rather than accept the losses as yet another consequence of official corruption and impunity, Mexican civil society rebelled. The people sustained protests, enlisted the support of international organizations, and exposed the government’s crimes and attempted cover-up. This study concludes that, despite the failure in the following years to locate the missing and bring the perpetrators to justice, the victims’ families and their allies demonstrated their power to hold the government to account and insist on attention to such violations. Ayotzinapa marks a watershed event in the history of the defense of human rights in Mexico

Designing a microbial cultivation platform for continuous biopharmaceutical production

The existing biopharmaceutical manufacturing paradigm is poorly suited to produce biologic drugs on demand at a point-of-care. Generally, commercial-scale (~2,000 - 10,000 L) manufacturing using fed-batch cultivation and fixed stainless-steel infrastructure is concentrated in developed nations and results in process cycle times on the order of weeks to months.1,2 Coupled with the complex logistical challenges associated with continuous “plant-to-patient” cold-chains, the geographically biased nature of therapeutic protein production today can limit access to biologic drugs in developing areas of the world.3 There is an opportunity to create technologies capable of rapidly generating biopharmaceuticals in situ in emergency situations, in remote healthcare settings, and in the battlefield. A platform that incorporates a modular suite of bioreactor, purification, and in-line analytics technologies has the potential to bridge this gap if developed in parallel with appropriately engineered stains of a flexible expression host. This poster will describe a multifaceted approach towards the development of a fully automated bench-scale perfusion process for the cultivation of Pichia pastoris and expression of therapeutically relevant heterologous proteins. We demonstrate the application of computational fluid dynamics (CFD) simulations to the optimization of the cultivation environment within our bench-top bioreactors. We further show that Pichia pastoris is amenable to secreting a variety of recombinant proteins spanning a range of preexisting drug classes (e.g. hormones, cytokines, monoclonal antibodies, vaccine antigens). Among these therapeutic proteins are molecules that require proper co-/post-translational processing for bioactivity. We envision that the development of P. pastoris strains with the capability to perform these critical processing steps in vivo will mitigate the need to chemically modify proteins post-expression and reduce the number of unit operations required in a typical upstream process

A perfusion-capable microfluidic bioreactor for assessing microbial heterologous protein production

We present an integrated microfluidic bioreactor for fully continuous perfusion cultivation of suspended microbial cell cultures. This system allowed continuous and stable heterologous protein expression by sustaining the cultivation of Pichia pastoris over 11 days. This technical capability also allowed testing the impact of perfusion conditions on protein expression. This advance should enable small-scale models for process optimization in continuous biomanufacturing.United States. Defense Advanced Research Projects Agency (N66001-13-C-4025)National Cancer Institute (U.S.) (P30-CA14051)United States. National Institutes of Health (2T32GM008334-26

Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris

Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, ‘hit-to-lead identification’ remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigenspecific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV).Bill & Melinda Gates FoundationUnited States. Defense Advanced Research Projects AgencySpace and Naval Warfare Systems Center San Diego (U.S.) (Contract N66001-13-C-4025)W. M. Keck FoundationNational Institute of Allergy and Infectious Diseases (U.S.) (U19AI090970).National Cancer Institute (U.S.) (David H. Koch Institute for Integrative Cancer Research at MIT. Support (Core) Grant P30-CA14051

Mathematical model of the dynamics of psychotherapy

The success of psychotherapy depends on the nature of the therapeutic relationship between a therapist and a client. We use dynamical systems theory to model the dynamics of the emotional interaction between a therapist and client. We determine how the therapeutic endpoint and the dynamics of getting there depend on the parameters of the model. Previously Gottman et al. used a very similar approach (physical-sciences paradigm) for modeling and making predictions about husband–wife relationships. Given that this novel approach shed light on the dyadic interaction between couples, we have applied it to the study of the relationship between therapist and client. The results of our computations provide a new perspective on the therapeutic relationship and a number of useful insights. Our goal is to create a model that is capable of making solid predictions about the dynamics of psychotherapy with the ultimate intention of using it to better train therapists

On-demand manufacturing of clinical-quality biopharmaceuticals

Conventional manufacturing of protein biopharmaceuticals in centralized, large-scale, single-product facilities is not well-suited to the agile production of drugs for small patient populations or individuals. Previous solutions for small-scale manufacturing are limited in both process reproducibility and product quality, owing to their complicated means of protein expression and purification. We describe an automated, benchtop, multiproduct manufacturing system, called Integrated Scalable Cyto-Technology (InSCyT), for the end-to-end production of hundreds to thousands of doses of clinical-quality protein biologics in about 3 d. Unlike previous systems, InSCyT includes fully integrated modules for sustained production, efficient purification without the use of affinity tags, and formulation to a final dosage form of recombinant biopharmaceuticals. We demonstrate that InSCyT can accelerate process development from sequence to purified drug in 12 weeks. We used integrated design to produce human growth hormone, interferon α-2b and granulocyte colony-stimulating factor with highly similar processes on this system and show that their purity and potency are comparable to those of marketed reference products

Principles of counseling and psychotherapy : learning the essential domains and nonlinear thinking of master practitioners/ Mozdzierz

xxiii, 474 hal.; 26 cm

Principles of counseling and psychotherapy : learning the essential domains and nonlinear thinking of master practitioners/ Mozdzierz

xxiii, 474 hal.; 26 cm

High temperature creep performance and microstructure of SiC-C-SiC composites

The high temperature creep behaviour of a 2D SiC/C/SiC composite is studied at temperatures ranging from 1200 to 1673K, under stresses from 50 to 500MPa. Due to the microstructural evolution of the material, steady state creep is never reached ; creep toughening is observed. Microcrack formation and propagation in the interface and in the matrix were identified as contributing to the deformation