Cell culture scale translation from a 24-well Single-Use miniature bioreactor and subsequent impact on product and broth quality

To accelerate cell culture process development, most companies have validated scale-down models of their pilot and manufacturing scale bioreactors. Advancing such mimics to even smaller scales requires the large scale engineering environment to be accurately recreated. Here we describe a single-use microwell methodology that accurately reproduces not only cell growth kinetics but also key attributes related to product quality and broth processability. The μ24 miniature bioreactor system enables system level control of agitation (by orbital shaking), with individual well control of pH, DO and temperature. Two distinct plate types are investigated, allowing for either headspace or direct gas sparging. An engineering characterisation was performed evaluating fluid mixing, gas transfer capacity and the dispersed gas phase. Cell culture is investigated using a model CHO DG44 cell line expressing a whole IgG1 mAb [1]. In addition, this work describes scale-up of μ24 results to conventional laboratory scale stirred tank bioreactors (2L) and use of the device for selection of robust and scaleable cell lines through evaluation of product quality. The ‘broth quality’ is also evaluated for primary clarification efficiency using an Ultra Scale-Down (USD) depth filtration rig that requires quantities of material compatible with those available from the miniature bioreactors. Apparent kLa values ranged between 3–22 hr-1 and 4–53 hr-1 for headspace aeration and direct gas sparging respectively. Mixing times were generally in the range 1–13 seconds and decreased with increasing shaking frequency (500–800 rpm). Direct gas sparging also helped to reduce tm values. Cultures performed with headspace aeration showed the highest VCD and antibody titres, whereas those operated with direct gas sparging showed cell growth kinetics and product titres that were more comparable to those found in a conventional 2L stirred bioreactor. Initial results also indicate that key product and broth processability attributes are maintained making the combination of μ24 and USD technologies useful tools in ‘Quality by Design’ driven cell culture process development

A critical review of Pongamia pinnata multiple applications: from land remediation and carbon sequestration to socioeconomic benefits

Pongamia pinnata (L.) Pierre (Pongamia) is a tree native to Southeast Asia. Recently, interest in Pongamia focused on its potential as a biofuel source as its seeds contain around 40% oil. However, Pongamia has multiple applications beyond biofuel production. It is a legume, can form symbiotic associations with mycorrhizal fungi, has been shown to be tolerant to drought, salinity, and heavy metals in soil, and has potential to mitigate climate change. Additionally, Pongamia oil has medicinal properties, can be used as biopesticide, insect repellent, to produce soap, and as a source of edible grade vegetable oil. The seed cake can be used as a source of bioenergy, food and feed protein, and organic fertiliser, and the flowers are a good source of pollen and nectar. Pongamia can also bring socio-economic benefits as its ability to restore degraded and contaminated land provides opportunities for local communities through novel valorisation pathways. These multiple applications have potential to form part of a circular bioeconomy in line with sustainable development goals. Although research on the multiple applications of Pongamia has grown considerably, knowledge gaps remain and these need to be addressed so that the full potential of Pongamia can be achieved. Further understanding of the mechanisms underlying its resilience to abiotic stresses, phytoremediation potential and biotic interactions should be a priority, and co-ordinated breeding efforts will be key. Here, we critically review the available literature on Pongamia and highlight gaps in knowledge in which future research should focus on to ensure that the full potential of this versatile tree can be achieved. We conclude that Pongamia can potentially form part of a circular bioeconomy and that harnessing the multiple applications of Pongamia in a holistic manner, with collaboration among key stakeholders, is crucial for the successful application of its benefits far beyond biofuel production

The GALNT9, BNC1 and CCDC8 genes are frequently epigenetically dysregulated in breast tumours that metastasise to the brain.

Tumour metastasis to the brain is a common and deadly development in certain cancers; 18-30 % of breast tumours metastasise to the brain. The contribution that gene silencing through epigenetic mechanisms plays in these metastatic tumours is not well understood

The relationship between occupational demands and well-being of performing artists: A systematic review

Background: Performing artists are exposed to a range of occupational demands from organisational, interpersonal and intrapersonal sources, which may impact their well-being. The aim of this systematic review was to evaluate and synthesise the literature where researchers have considered the relationship between occupational demands and well-being in performing artists. Methods: A mixed-methods systematic review was conducted including professional and student performing artists. Quantitative, qualitative and mixed-methods study designs were eligible for inclusion in the review. A total of 14 databases were searched from their inception through to October 2017, including MEDLINE, EMBASE and Scopus. Critical appraisal was conducted using the Mixed-Methods Appraisal Tool and results presented as a narrative synthesis. Results: A total of 20 studies were included in the review, comprising of quantitative (n=7), qualitative (n=9) and mixed-methods (n=4) study designs. Several frameworks of occupational stress and well-being were explored in relation to the results. Organisational, social and emotional demands were associated with lower well-being. Conversely, music-making, performance activities and social support were reported to be resources and were related to higher well-being. Conclusion: This systematic review highlights the need for researchers in this field to adopt methodologically robust study designs, which are informed by appropriate theoretical frameworks. The paucity of high quality and theoretically informed research in this area is a hindrance to the development of evidence-based interventions for this population

Field-Caught Permethrin-Resistant Anopheles gambiae Overexpress CYP6P3, a P450 That Metabolises Pyrethroids

Insects exposed to pesticides undergo strong natural selection and have developed various adaptive mechanisms to survive. Resistance to pyrethroid insecticides in the malaria vector Anopheles gambiae is receiving increasing attention because it threatens the sustainability of malaria vector control programs in sub-Saharan Africa. An understanding of the molecular mechanisms conferring pyrethroid resistance gives insight into the processes of evolution of adaptive traits and facilitates the development of simple monitoring tools and novel strategies to restore the efficacy of insecticides. For this purpose, it is essential to understand which mechanisms are important in wild mosquitoes. Here, our aim was to identify enzymes that may be important in metabolic resistance to pyrethroids by measuring gene expression for over 250 genes potentially involved in metabolic resistance in phenotyped individuals from a highly resistant, wild A. gambiae population from Ghana. A cytochrome P450, CYP6P3, was significantly overexpressed in the survivors, and we show that the translated enzyme metabolises both alpha-cyano and non–alpha-cyano pyrethroids. This is the first study to demonstrate the capacity of a P450 identified in wild A. gambiae to metabolise insecticides. The findings add to the understanding of the genetic basis of insecticide resistance in wild mosquito populations

Study Protocol - Accurate assessment of kidney function in Indigenous Australians: aims and methods of the eGFR Study

Background: There is an overwhelming burden of cardiovascular disease, type 2 diabetes and chronic kidney disease among Indigenous Australians. In this high risk population, it is vital that we are able to measure accurately kidney function. Glomerular filtration rate is the best overall marker of kidney function. However, differences in body build and body composition between Indigenous and non-Indigenous Australians suggest that creatinine-based estimates of glomerular filtration rate derived for European populations may not be appropriate for Indigenous Australians. The burden of kidney disease is borne disproportionately by Indigenous Australians in central and northern Australia, and there is significant heterogeneity in body build and composition within and amongst these groups. This heterogeneity might differentially affect the accuracy of estimation of glomerular filtration rate between different Indigenous groups. By assessing kidney function in Indigenous Australians from Northern Queensland, Northern Territory and Western Australia, we aim to determine a validated and practical measure of glomerular filtration rate suitable for use in all Indigenous Australians

Job Characteristics and Labour Supply

We document the importance of non-pecuniary aspects in employment relationships by showing that labour supply elasticities differ significantly among individuals' job characteristics. Factor analysis indicates the relevance of four characteristics: autonomy, workload, variety and job security. Using a discrete choice model of family labour supply on the basis of Australian data, we show that income elasticities are significantly higher among individuals with 'good' characteristics (e.g. a securer job) whereas wage elasticities are significantly lower. This result holds for both men and women. Our main hypothesis are derived within the 'new approach to consumer theory proposed by Lancaster.Wir untersuchen den Einfluss von (nicht-monetären) Arbeitsbedingungen auf individuelle Arbeitsangebotsentscheidungen mittels eines diskreten Entscheidungsmodells. Um Arbeitsbedingungen zunächst zu konzeptionalisieren, greifen wir auf den sog. 'Characteristics Approach' zurück. Die Wirkung von verschiedenen Charakteristika eines Jobs wie z.B. der Grad an Autonomie oder die Arbeitsauslastung auf das individuelle Arbeitsangebot lassen sich dadurch sowohl theoretisch als auch empirisch in einem konsistenten Analyserahmen untersuchen. Zum einen zeigen wir, dass man theoretisch einen positiven Zusammenhang zwischen einer Verbesserung der Arbeitseigenschaften und der tatsächlich gearbeiteten Arbeitsstundenzahl erwarten würde. Zum anderen weisen unsere Befunde darauf hin, dass sich dieser theoretische Zusammenhang, auch in Form von signifikant unterschiedlichen Arbeitsangebotselastizitäten zwischen solchen Individuen mit guten Arbeitseigenschaften und jenen mit weniger vorteilhaften, niederschlägt