Integrated scalable cyto-technology for recombinant protein bioprocessing

Biological knowledge of infectious diseases and other diseases for which vaccines may provide therapeutic benefits, such as cancer, is growing at an accelerated pace. The implications of this knowledge are improved stratification of diseases, possibilities for personalized treatments, and explicit understanding of protective immune responses to be elicited by vaccines. With this knowledge, it is becoming increasingly feasible to engineer vaccines for specific responses rather than relying on empirical development. Despite this potential, the challenge of routine, low-cost manufacturing of vaccines creates a barrier to transforming health care in both high- and low-resource countries. Vaccines today do not benefit from well-defined, platform-like processes for manufacturing, and concepts such as continuous bioprocessing remain largely within the realm of biopharmaceutical products. The InSCyT platform is an advanced prototype manufacturing system that provides integrated and automated production and purification of multiple protein therapeutics. The system allows end-to-end manufacturing of 100\u27s to 10,000\u27s of doses of recombinant protein drugs in days. It uses a state-of-the-art approach to process design and implementation that takes advantage of a fast-growing, tractable microbial host (Pichia pastoris) and continuous processing for automated, hands-free purification through simple 2- or 3-stage chromatographic processes. The platform design is highly modular, allowing facile process development and process deployment for multiple products. This feature emerges from the predictable behavior of the fermentation and cell culture fluids, and rapid cloning of new molecules, that together facilitate fast development of entirely new processes in weeks. To date, this system has been used to reproducibly manufacture high-quality human growth hormone (hGH), granulocyte-colony stimulating factor (G-CSF), and interferon-α2b (IFN-α2b) in an integrated, automated manner. The speed of production using the InSCyT prototype allows volumetric productivities that compare favorably to those for mammalian-based production. This talk will outline the design and capabilities of the InSCyT system, demonstrate the quality of biologic drugs made to date on the system, and outline opportunities for advancing the platform to provide new capabilities in manufacturing recombinant proteins for use in vaccines. As part of a Gates Foundation-funded Grand Challenge called ULTRA, we have begun to assess the feasibility of manufacturing millions of doses of a trivalent recombinant rotavirus vaccine annually on a small-scale production system like InSCyT. Integrated bioprocessing enabled by systems such as these could offer potential advantages for routine production in local regions with minimal infrastructure, and for democratization of manufacturing capacity for new vaccine concepts and personalized treatments in cancer

Insights into sugar metabolism during bilberry (Vaccinium myrtillus L.) fruit development

Bilberry fruit is regarded as one of the best natural sources of anthocyanins and iswidely explored for its health-beneficial compounds. Besides anthocyanins, one of themajor attributes that determine the berry quality is the accumulation of sugars thatprovide sweetness and flavor to ripening fruit. In this study, we have identified 25 sugarmetabolism-related genes in bilberry, including invertases (INVs), hexokinases (HKs),fructokinases (FKs), sucrose synthases (SSs), sucrose phosphate synthases (SPSs), andsucrose phosphate phosphatases (SPPs). The results indicate that isoforms of the identified genes are expressed differentially during berry development, suggesting special-ized functions. The highest sugar content was found in ripe berries, with fructose andglucose dominating accompanied by low sucrose amount. The related enzyme activi-ties during berry development and ripening were further analyzed to understand themolecular mechanism of sugar accumulation. The activity of INVs in the cell wall andvacuole increased toward ripe berries. Amylase activity involved in starch metabolismwas not detected in unripe berries but was found in ripe berries. Sucrose resynthesizing SS enzyme activity was detected upon early ripening and had the highestactivity in ripe berries. Interestingly, our transcriptome data showed that supplementalirradiation with red and blue light triggered upregulation of several sugar metabolism-related genes, including α- and β-amylases. Also, differential expression patterns inresponses to red and blue light were found across sucrose, galactose, and sugar-alcoholmetabolism. Our enzymological and transcriptional data provide new understanding ofthe bilberry fruit sugar metabolism having major effect on fruit quality (20) (PDF) Insights into sugar metabolism during bilberry ( Vaccinium myrtillus L.) fruit development

Comparing Service Delivery Models for Children with Developmental Delays in Canada: Adaptive and Maladaptive Behaviours, Parental Perceptions of Stress and of Care

This study compares two service delivery models (community-based and centre-based), examining them in light of children’s adaptive and maladaptive behaviours, and parental perceptions of stress and of care. More specifically, parents of 96 children with developmental delays assessed their children’s adaptive and maladaptive behaviours and rated their own perceived levels of stress as well as their perceptions of care from service providers. Findings indicated that children from the community-based sites were perceived as having less severe social skill deficits than those from centre-based sites. Regarding parental stress, mothers from community-based settings reported more challenges with their child’s father than did the mothers from centre-based settings; and fathers from the community-based settings reported more challenges related to their health than did the fathers from the centre-based settings. Regarding care, parents from the centre-based settings had more positive perceptions of care than did parents from the community-based settings. Therefore, in general, parents receiving services within community-based settings reported fewer positive perceptions of care and more challenges than those from centre-based settings. Overall, the results of this investigation can inform future programming for community- and centre-based service delivery systems. More specifically, the findings highlight the important role that family-centred care can play in supporting the needs of children with developmental delays and their families; particularly for families using community-based services

Influence of Arctic light conditions on crop production and quality

The natural light conditions above the Arctic Circle are unique in terms of annual variation creating special growth conditions for crop production. These include low solar elevations, very long daily photosynthetic light periods, midnight sun/absence of dark nights, and altered spectral distribution depending on solar elevation. All these factors are known to affect the growth and the metabolism of plants, although their influence on northern crop plants has not yet been reviewed. The ongoing global warming is especially affecting the temperature x light interactions in the Arctic, and understanding the impact on crop production and plant metabolism will be important for an Arctic contribution to global food production. Arctic light conditions have a strong influence on the timing of plant development, which together with temperature limits the number of cultivars suitable for Arctic agriculture. This review compiles information from the reports about the effects of light conditions at high latitudes on growth, biomass production, flowering and quality of the crop plants and discusses the gained knowledge and the key gaps to be addressed.Peer reviewe

A preliminary study of genetic factors that influence susceptibility to bovine tuberculosis in the British cattle herd

Associations between specific host genes and susceptibility to Mycobacterial infections such as tuberculosis have been reported in several species. Bovine tuberculosis (bTB) impacts greatly the UK cattle industry, yet genetic predispositions have yet to be identified. We therefore used a candidate gene approach to study 384 cattle of which 160 had reacted positively to an antigenic skin test (‘reactors’). Our approach was unusual in that it used microsatellite markers, embraced high breed diversity and focused particularly on detecting genes showing heterozygote advantage, a mode of action often overlooked in SNP-based studies. A panel of neutral markers was used to control for population substructure and using a general linear model-based approach we were also able to control for age. We found that substructure was surprisingly weak and identified two genomic regions that were strongly associated with reactor status, identified by markers INRA111 and BMS2753. In general the strength of association detected tended to vary depending on whether age was included in the model. At INRA111 a single genotype appears strongly protective with an overall odds ratio of 2.2, the effect being consistent across nine diverse breeds. Our results suggest that breeding strategies could be devised that would appreciably increase genetic resistance of cattle to bTB (strictly, reduce the frequency of incidence of reactors) with implications for the current debate concerning badger-culling

Red and blue light treatments of ripening bilberry fruits reveal differences in signalling through abscisic acid-regulated anthocyanin biosynthesis

The biosynthesis of anthocyanins has been shown to be influenced by light quality. However, the molecular mechanisms underlying the light-mediated regulation of fruit anthocyanin biosynthesis are not well understood. In this study, we analysed the effects of supplemental red and blue light on the anthocyanin biosynthesis in non-climacteric bilberry (Vaccinium myrtillus L.). After 6 days of continuous irradiation during ripening, both red and blue light elevated concentration of anthocyanins, up to 12- and 4-folds, respectively, compared to the control. Transcriptomic analysis of ripening berries showed that both light treatments up-regulated all the major anthocyanin structural genes, the key regulatory MYB transcription factors and abscisic acid (ABA) biosynthetic genes. However, higher induction of specific genes of anthocyanin and delphinidin biosynthesis alongside ABA signal perception and metabolism were found in red light. The difference in red and blue light signalling was found in 9-cis-epoxycarotenoid dioxygenase (NCED), ABA receptor pyrabactin resistance-like (PYL) and catabolic ABA-8'hydroxylase gene expression. Red light also up-regulated expression of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) domain transporters, which may indicate involvement of these proteins in vesicular trafficking of anthocyanins during fruit ripening. Our results suggest differential signal transduction and transport mechanisms between red and blue light in ABA-regulated anthocyanin and delphinidin biosynthesis during bilberry fruit ripening.Peer reviewe