Temperature Controlled Packaging Container for Biologics and Pharmaceuticals

Clinical Supplies Management, Inc. (CSM) provides clinical trial services to biotechnology and pharmaceutical companies by shipping pharmaceuticals to clinics and other patients. In winter 2014, CSM presented a temperature control packaging project to the Cal Poly Senior Project class. As a result, three Cal Poly Mechanical Engineering Students were tasked to design, manufacture, and qualify a shipping container that would maintain ten 10 mL vials between 2-8°C for 96 hours. The final product would have to cost less to ship than the Credo Cube, CSM’s current temperature control packaging product. After considerable research and analysis on possible temperature control technologies, it was determined that the most reliable and economical solution was similar technology to the Credo Cube. The main features of the product are as follows: Vacuum Insulated Paneling (VIP) is used as a thermal resistor to reduce the heat transfer from the surroundings to the payload Phase change material (PCM) maintains the desired payload temperature An optimization for the specified payload was performed by creating designs to reduce the tare weight, and dimensional weight using the above technologies. During the manufacturing phase, the selected vendor for phase change material was unwilling to accommodate a lead time within the senior project’s scope. This was believed to be partially due to the low volume and direct competition of the potential product. CSM was in contact with the vendor, and it was decided that the legal steps needed to move forward with the selected PCM would be too far outside of the project’s scope. Consequently, it was decided between the project team and CSM that using water as a prototype PCM was acceptable. Instead of a final product, a proof of concept prototype and a calculator that would help CSM redesign the product if other VIP or PCM were to be pursued in the future acted as the final deliverables. Although the prototype was not able to maintain temperature within range for 96 hours, the design can easily be scaled to do so with further testing and the use of the provided calculator. The following report details the design, manufacture, and qualification of the shipping container prototype

Preparation of highly and generally enriched mammalian tissues for solid state NMR.

An appreciable level of isotope labelling is essential for future NMR structure elucidation of mammalian biomaterials, which are either poorly expressed, or unexpressable, using micro-organisms. We present a detailed protocol for high level (13)C enrichment even in slow turnover murine biomaterials (fur keratin), using a customized diet supplemented with commercial labelled algal hydrolysate and formulated as a gel to minimize wastage, which female mice consumed during pregnancy and lactation. This procedure produced approximately eightfold higher fur keratin labelling in pups, exposed in utero and throughout life to label, than in adults exposed for the same period, showing both the effectiveness, and necessity, of this approach.The authors would like to acknowledge funding from the Biotechnology and Biological Sciences Research Council for DGR and RR; Engineering and Physical Sciences Research Council for WYC and VWCW; National Institute of Health Research for RAB.This is the final version of the article. It first appeared from Springer via http://dx.doi.org/10.1007/s10858-015-9977-

Proline provides site-specific flexibility for in vivo collagen.

Fibrillar collagens have mechanical and biological roles, providing tissues with both tensile strength and cell binding sites which allow molecular interactions with cell-surface receptors such as integrins. A key question is: how do collagens allow tissue flexibility whilst maintaining well-defined ligand binding sites? Here we show that proline residues in collagen glycine-proline-hydroxyproline (Gly-Pro-Hyp) triplets provide local conformational flexibility, which in turn confers well-defined, low energy molecular compression-extension and bending, by employing two-dimensional 13C-13C correlation NMR spectroscopy on 13C-labelled intact ex vivo bone and in vitro osteoblast extracellular matrix. We also find that the positions of Gly-Pro-Hyp triplets are highly conserved between animal species, and are spatially clustered in the currently-accepted model of molecular ordering in collagen type I fibrils. We propose that the Gly-Pro-Hyp triplets in fibrillar collagens provide fibril "expansion joints" to maintain molecular ordering within the fibril, thereby preserving the structural integrity of ligand binding sites.BBSRC, EPSRC, Raymond and Beverly Sackler Fund for Physics of Medicine, Wellcome Trust, ER

NMR spectroscopy of native and in vitro tissues implicates polyADP ribose in biomineralization.

Nuclear magnetic resonance (NMR) spectroscopy is useful to determine molecular structure in tissues grown in vitro only if their fidelity, relative to native tissue, can be established. Here, we use multidimensional NMR spectra of animal and in vitro model tissues as fingerprints of their respective molecular structures, allowing us to compare the intact tissues at atomic length scales. To obtain spectra from animal tissues, we developed a heavy mouse enriched by about 20% in the NMR-active isotopes carbon-13 and nitrogen-15. The resulting spectra allowed us to refine an in vitro model of developing bone and to probe its detailed structure. The identification of an unexpected molecule, poly(adenosine diphosphate ribose), that may be implicated in calcification of the bone matrix, illustrates the analytical power of this approach

May Measurement Month 2017: an analysis of blood pressure screening results from Australia - South-East Asia and Australasia

Increased blood pressure (BP) is the single biggest contributing risk factor to the global disease burden. May Measurement Month (MMM) is a global initiative of the International Society of Hypertension aimed at raising awareness of high BP. In Australia, hypertension affects around six million adults and continues to remain the greatest attributable cause of cardiovascular mortality and morbidity (48.3%), stroke deaths (28%), and kidney disease (14%). An opportunistic cross-sectional survey was carried out during May 2017 predominantly in capital cities across Australia which included adult volunteers. Blood pressure measurement, the definition of hypertension and statistical analysis followed the standard MMM protocol. Additional information obtained included anthropometric data and responses to questionnaires on demographic, lifestyle, and environmental factors. Data were collected from 3817 individuals. After multiple imputation, of the 3758 individuals for whom a mean of the second and third BP reading was available, 1188 (31.2%) had hypertension. Of 3213 individuals not receiving antihypertensive treatment, 591 (18.4%) were hypertensive, and 239 (40.1%) of the 596 individuals receiving treatment had uncontrolled BP. Adjusted BP was higher in association with antihypertensive medication, cerebrovascular disease, smoking, and alcohol consumption. Blood pressure was higher when measured on the right arm and on Tuesdays. MMM17 was one of the largest BP screening campaigns undertaken in Australia using standardized BP measurements. In line with previous surveys, around one-third of screened adults had hypertension and approximately 40% of treated individuals remained uncontrolled. These results suggest that opportunistic screening can identify significant numbers with raised BP

The genome sequence of the Norway rat, Rattus norvegicus Berkenhout 1769.

We present a genome assembly from an individual male Rattus norvegicus (the Norway rat; Chordata; Mammalia; Rodentia; Muridae). The genome sequence is 2.44 gigabases in span. The majority of the assembly is scaffolded into 20 chromosomal pseudomolecules, with both X and Y sex chromosomes assembled. This genome assembly, mRatBN7.2, represents the new reference genome for R. norvegicus and has been adopted by the Genome Reference Consortium

Pigmentation Chemistry and Radical-Based Collagen Degradation in Alkaptonuria and Osteoarthritic Cartilage

Alkaptonuria (AKU) is a rare disease characterized by high levels of homogentisic acid (HGA); patients suffer from tissue ochronosis: dark brown pigmentation, especially of joint cartilage, leading to severe early osteoarthropathy. No molecular mechanism links elevated HGA to ochronosis; the pigment's chemical identity is still not known, nor how it induces joint cartilage degradation. Here we give key insight on HGA-derived pigment composition and collagen disruption in AKU cartilage. Synthetic pigment and pigmented human cartilage tissue both showed hydroquinone-resembling NMR signals. EPR spectroscopy showed that the synthetic pigment contains radicals. Moreover, we observed intrastrand disruption of collagen triple helix in pigmented AKU human cartilage, and in cartilage from patients with osteoarthritis. We propose that collagen degradation can occur via transient glycyl radicals, the formation of which is enhanced in AKU due to the redox environment generated by pigmentation

Demand for Genetic Resources and the U.S. National Plant Germplasm System

Although genetic resources have strong public-goods characteristics, public genebanks often have struggled for adequate funding. A review of economic literature on the value of plant genetic resources indicated that more information is needed about germplasm use. The data compiled in this paper examine patterns of germplasm use for one of the world\u27s largest national genebank networks, the U.S. National Plant Germplasm System (NPGS). Data on 10 major crops, gathered directly from within the NPGS and from end-users, revealed patterns of usage for germplasm during the period from 1995 to 1999. Data were collected describing the characteristics of NPGS users, the types of germplasm requested, the purpose of requests, and, when applicable, the specific traits sought. From these findings, we estimated the utility of distributed materials, their secondary use, and projected future demand for NPGS resources. To explore relationships between the usefulness of germplasm samples and accompanying data in a more systematic fashion, we estimated a linear regression. The regression model suggests that accompanying data make germplasm more useful. We conclude that demand for NPGS resources was substantial and came from broad range of users. Utilization rates were higher than suggested by past studies. Countries with developing economies made greater use of NPGS resources, relatively speaking, than did countries with high-income economies. Finally, demand for NPGS resources is likely to increase, especially among users in countries with developing economies