NEW GENERATION CO-OPERATIVES AND THE LAW IN SASKATCHEWAN

Agribusiness,

NEW GENERATION CO-OPERATIVES: RESPONDING TO CHANGES IN AGRICULTURE

Agribusiness,

Development of CRISPR/Cas9 in vivo bladder cancer models

Bladder cancer prevalence is high, and patients diagnosed with this malignancy incur a high economic burden and a poor-quality lifestyle. The majority of bladder cancers are urothelial carcinomas, with two subtypes: papillary non-invasive and muscle-invasive bladder cancer. The muscle-invasive subtype is associated with a poor prognosis and a high mutation frequency. Nevertheless, there are no efficient in vivo bladder cancer models in the field to study disease initiation and progression. Thus far, most in vivo bladder cancer models rely on the Cre-LoxP system. However, this technique is expensive and time-consuming. Furthermore, it depends on available mice with floxed alleles for target genes. Recently, CRISPR/Cas9 has been used to study cancer and overcomes the hurdles of Cre-LoxP models. The objective of this work is to determine if CRISPR/Cas9 can be coopted for bladder cancer research in vivo. As-proof-of concept CRISPR/Cas9 was used to recapitulate a Cre-LoxP muscle-invasive bladder mouse model produced by knockout of tumor suppressors PTEN and TP53. Single guide RNAs targeting TP53 and PTEN were delivered into the bladder urothelium of Cas9 expressing mice by a novel electroporation approach. Histological and pathological characterization of bladders indicated tumor development in 2 mice, with a tumor penetrance of 33%. Tumors phenotypically resembled hyperplasia and papillary carcinoma, with an increase of CK5 positive basal layer. Papillary tumors exhibited an increase in cell proliferation compared to controls. Overall, optimizations are required in order to use the CRISPR/Cas9 technique for in vivo bladder cancer models

New Generation Cooperatives and Cooperative Theory

North Dakota and Minnesota are currently witnessing a renewal in the growth of cooperative enterprises. At the heart of this renewal lies the so-called New Generation Cooperative (NGC), a term that has been applied to the dozens of value-added processing, selected membership cooperatives that have recently formed in the region's agricultural sector. A key organizational feature of NGCs is the linking of producer capital contributions and product delivery rights. This paper describes the organizational features of NGCs and positions them in the broader context of cooperative incentive structures, governance structures, and the cooperative development process. More generally, the paper uses NGCs as a lens through which important elements of cooperative theory can be reviewed

Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation.

Genetically engineered mouse models (GEMMs) are extremely valuable in revealing novel biological insights into the initiation and progression mechanisms of human diseases such as cancer. Transgenic and conditional knockout mice have been frequently used for gene overexpression or ablation in specific tissues or cell types in vivo. However, generating germline mouse models can be time-consuming and costly. Recent advancements in gene editing technologies and the feasibility of delivering DNA plasmids by viral infection have enabled rapid generation of non-germline autochthonous mouse cancer models for several organs. The bladder is an organ that has been difficult for viral vectors to access, due to the presence of a glycosaminoglycan layer covering the urothelium. Here, we describe a novel method developed in lab for efficient delivery of DNA plasmids into the mouse bladder urothelium in vivo. Through intravesical instillation of pCAG-GFP DNA plasmid and electroporation of surgically exposed bladder, we show that the DNA plasmid can be delivered specifically into the bladder urothelial cells for transient expression. Our method provides a fast and convenient way for overexpression and knockdown of genes in the mouse bladder, and can be applied to building GEMMs of bladder cancer and other urological diseases

FOST 2 Upgrade with Hollow-Fiber CTA FO Module and Generation of Osmotic Agent for Microorganism Growth Studies

FOST 2 is an integrated membrane system that incorporates a forward osmosis subsystem and a reverse osmosis subsystem working in series. It has been designed as a post treatment system to process the effluent from the Membrane Aerated Biological Reactor developed at NASA Johnson Space Center and Texas Tech University. Its function is to remove dissolved solids residual such as ammonia and suspended solids, as well as to provide a physical barrier to microbial and viral contamination. A tubular CTA membrane module from HTI and a flat-sheet lipid-base membrane module from Porifera were integrated and tested on FOST 2 in the past, using both a bioreactor's effluent and greywater as the feed solution. This paper documents the performance of FOST 2 after its upgrade with a hollow-fiber CTA membrane module from Toyobo, treating real black-water to generate the osmotic agent solution necessary to conduct growth studies of genetically engineered microorganism for the Synthetic Biological Membrane project

The Effect of Fatty Acids to Protect Forward Osmosis Membranes from Damage

NASA has conducted research and development on forward osmosis (FO) membranes for wastewater reclamation in space since 1993. The lessons learned during operation of the International Space Station and FO based technologies on the ground taught us that reliability is a key limitation. Membranes are susceptible to organic fouling, oxidation and calcium scaling, and these factors tend to damage the membrane reducing their operating life and performance. The development of a Synthetic Biological Membrane (SBM), a membrane that mimics naturally occurring biological processes, will mitigate membrane damage and improve reliability. The SBM is a lipid-based membrane with a protective fatty acid layer configured for use in a FO water purification system. In this configuration, the protective layer on the surface of the lipid membrane is composed of fatty acids (FA). The FA interact with the chemicals found in the wastewater feed, and protect the membrane from damage. In this study, we conducted preliminary experiments to determine the feasibility of using fatty acids to alleviate damage from calcium scaling, oxidation and organic fouling