A rapid and inexpensive method for surface sterilisation of Ecklonia radiata (Phaeophyta) for tissue culture

Sections of the thallus of the brown alga Ecklonia radiata (C. Ag.) J. Ag. were surface sterilised for tissue culture by dipping in 70% ethanol for 30 s, followed by sterile deionised water for 30 min. A high percentage of aseptic viable explants could be obtained using this treatment, and growth in vitro was initiated

Tissue culture of brown seaweed

The harvesting of macrophyte algae (seaweeds) is well established and on a world-wide scale more than 180,000 tonnes dry weight of algae such as the phaeophytes Laminaria, Undaria,Sargassum and Macrocystis, the red algae Eucheuma, Gracilaria and Porphyra, and the green algae Ulva, Monostroma and Caulerpa are harvested annually. Much of this algal biomass comes from farmed rather than wild species. The red and brown algae are the source of the phycocolloids agar, alginate, agarose and carrageenan which are of fundamental importance to the development of biotechnology; i.e. for the culture of microorganisms (agar), for the separation of biomolecules (alginate and agarose) and for the production of food products (agar, carrageenan, alginate). The successful large-scale cultivation of these algae requires, amongst other things, the ability to select fast growing and disease resistant strains which produce large quantities of the desired phycocolloid. To this purpose classical plant breeding programs are being carried out, however these are slow and the production of superior cultivars takes much time and effort (Van der Meer 1988). In recent years there has therefore been much interest in developing protoplast and tissue culture systems which would allow more rapid selection and propagation of suitable cell lines, the possibility of producing hybrids by cell fusion and new strains by genetic engineering (Polne-Fuller and Gibor 1987b, Le Gallet al. 1990). Work in our laboratory has concentrated mainly on the brown algal genera Ecklonia and Cystophora. Ecklonia was chosen because it is easily obtained and is a potential source of alginate, and Cystophora, a genus endemic to Australia and New Zealand, because previous studies indicated that this genus appears to be a good source of tocopherols and tocotrienols (Gregson et al. 1977, Kazlauskas et al. 1981, unpubl. results). The tocopherols are of interest and also provide a convenient model for the study of the production of secondary metabolites in algal tissue culture. In this paper we describe some of our findings on the tissue culture of these species and on their tocopherol content

Three-dimensional numerical model of heat losses from district heating network pre-insulated pipes buried in the ground

The purpose of the paper is to investigate the challenges in modelling the energy losses of heating networks and to analyse the factors that influence them. The verification of the simulation was conducted on a test stand in-situ and based on the measurements of the testing station, a database for the final version of the numerical model was developed and a series of simulations were performed. Examples of the calculated results are shown in the graphs. The paper presents an innovative method of identify the energy losses of underground heating network pipelines and quantify the temperature distribution around them, in transient working conditions. The presented method makes use of numerical models and measured data of actual objects.The dimensions of the pipelines used were 6m wide, 8m high and 1m in depth, while they were simulated under conditions of zero heat flow in the ground, in the perpendicular to the sides direction of the calculated area and considering the effects of ground's thermal conductivity. The mesh was developed using advanced functions, which resulted its high quality with the average orthogonal quality of 0.99 (close to 1.00) and Skewness of 0.05 (between 0.00 and 0.25). To achieve better accuracy of the simulation model, the initial conditions were determined based on the numerical results of a three-dimensional analysis of heat losses, in steady state conditions in a single moment. The validation process confirmed the high quality of the model, as the differences between the ground temperatures were approximately 0.1°C

Reaching the unreached: de-mystifying the role of ICT in the process of doctoral research

Information and Communication Technology (ICT) has become a necessary element of academic practice in higher education today. Under normal circumstances, PhD students from all disciplines have to use ICT in some form throughout the process of their research, including the preparation, fieldwork, analysis and writing phases of their studies. Nevertheless, there has been little research to date that explores PhD students’ first-hand experiences of using various ICT to support their research practices. This paper brings together the findings and the key points from a review of significant parts of the existing literature associated with the role played by ICT in the processes PhD students use in doctoral research. The review is based on 27 papers appearing in international peer-reviewed journals published from 2005 to 2014. The study seeks to address the under-researched area in the current literature of how ICT plays a role in the processes of doctoral research. While there are many contributions taking the ‘institutional’ or ‘teaching’ perspectives, papers focusing on ‘student’ perspective, or the viewpoint of engaging ICT in daily study routine, are relatively fewer. As far as research methodology is concerned, this review found that many of the papers that were examined were mostly based on perception data such as surveys or interviews, while actual practice data were rarely present. With their ready access to technologies, PhD students are well positioned to take advantage of a range of technologies in order to carry out their research efficiently (in terms of means to an end) and effectively (in terms of reaching goals within a task). This review reveals that in the literature, this important area is under-represented

The Mitochondrial Apoptotic Effectors BAX/BAK Activate Caspase-3 and -7 to Trigger NLRP3 Inflammasome and Caspase-8 Driven IL-1beta Activation

Published: November 27, 2018Intrinsic apoptosis resulting from BAX/BAK-mediated mitochondrial membrane damage is regarded as immunologically silent. We show here that in macrophages, BAX/BAK activation results in inhibitor of apoptosis (IAP) protein degradation to promote caspase-8-mediated activation of IL-1β. Furthermore, BAX/BAK signaling induces a parallel pathway to NLRP3 inflammasome-mediated caspase-1-dependent IL-1β maturation that requires potassium efflux. Remarkably, following BAX/BAK activation, the apoptotic executioner caspases, caspase-3 and -7, act upstream of both caspase-8 and NLRP3-induced IL-1β maturation and secretion. Conversely, the pyroptotic cell death effectors gasdermin D and gasdermin E are not essential for BAX/BAK-induced IL-1β release. These findings highlight that innate immune cells undergoing BAX/BAK-mediated apoptosis have the capacity to generate pro-inflammatory signals and provide an explanation as to why IL-1β activation is often associated with cellular stress, such as during chemotherapy.James E. Vince, Dominic De Nardo, Wenqing Gao, Angelina J. Vince, Cathrine Hall, Kate McArthur, Daniel Simpson, Swarna Vijayaraj, Lisa M. Lindqvist, Philippe Bouillet, Mark A. Rizzacasa, Si Ming Man, John Silke, Seth L. Masters, Guillaume Lessene, David C.S. Huang, Daniel H.D. Gray, Benjamin T. Kile, Feng Shao, and Kate E. Lawlo

Assessment of systemic AAV-microdystrophin gene therapy in the GRMD model of Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD) is a progressive muscle wasting disease caused by the absence of dystrophin, a membrane-stabilizing protein encoded by the DMD gene. Although mouse models of DMD provide insight into the potential of a corrective therapy, data from genetically homologous large animals, such as the dystrophin-deficient golden retriever muscular dystrophy (GRMD) model, may more readily translate to humans. To evaluate the clinical translatability of an adeno-associated virus serotype 9 vector (AAV9)–microdystrophin (μDys5) construct, we performed a blinded, placebo-controlled study in which 12 GRMD dogs were divided among four dose groups [control, 1 × 1013 vector genomes per kilogram (vg/kg), 1 × 1014 vg/kg, and 2 × 1014 vg/kg; n = 3 each], treated intravenously at 3 months of age with a canine codon-optimized microdystrophin construct, rAAV9-CK8e-c-μDys5, and followed for 90 days after dosing. All dogs received prednisone (1 milligram/kilogram) for a total of 5 weeks from day-7 through day 28. We observed dose-dependent increases in tissue vector genome copy numbers; μDys5 protein in multiple appendicular muscles, the diaphragm, and heart; limb and respiratory muscle functional improvement; and reduction of histopathologic lesions. As expected, given that a truncated dystrophin protein was generated, phenotypic test results and histopathologic lesions did not fully normalize. All administrations were well tolerated, and adverse events were not seen. These data suggest that systemically administered AAV-microdystrophin may be dosed safely and could provide therapeutic benefit for patients with DMD

Associations of autozygosity with a broad range of human phenotypes

In many species, the offspring of related parents suffer reduced reproductive success, a phenomenon known as inbreeding depression. In humans, the importance of this effect has remained unclear, partly because reproduction between close relatives is both rare and frequently associated with confounding social factors. Here, using genomic inbreeding coefficients (F-ROH) for >1.4 million individuals, we show that F-ROH is significantly associated (p <0.0005) with apparently deleterious changes in 32 out of 100 traits analysed. These changes are associated with runs of homozygosity (ROH), but not with common variant homozygosity, suggesting that genetic variants associated with inbreeding depression are predominantly rare. The effect on fertility is striking: F-ROH equivalent to the offspring of first cousins is associated with a 55% decrease [95% CI 44-66%] in the odds of having children. Finally, the effects of F-ROH are confirmed within full-sibling pairs, where the variation in F-ROH is independent of all environmental confounding.Peer reviewe

Mild-to-Moderate Kidney Dysfunction and Cardiovascular Disease: Observational and Mendelian Randomization Analyses

BACKGROUND: End-stage renal disease is associated with a high risk of cardiovascular events. It is unknown, however, whether mild-to-moderate kidney dysfunction is causally related to coronary heart disease (CHD) and stroke. METHODS: Observational analyses were conducted using individual-level data from 4 population data sources (Emerging Risk Factors Collaboration, EPIC-CVD [European Prospective Investigation into Cancer and Nutrition-Cardiovascular Disease Study], Million Veteran Program, and UK Biobank), comprising 648 135 participants with no history of cardiovascular disease or diabetes at baseline, yielding 42 858 and 15 693 incident CHD and stroke events, respectively, during 6.8 million personyears of follow-up. Using a genetic risk score of 218 variants for estimated glomerular filtration rate (eGFR), we conducted Mendelian randomization analyses involving 413 718 participants (25917 CHD and 8622 strokes) in EPIC-CVD, Million Veteran Program, and UK Biobank. RESULTS: There were U-shaped observational associations of creatinine-based eGFR with CHD and stroke, with higher risk in participants with eG FR values 105 mL.min(-1).1.73 m(-2), compared with those with eG FR between 60 and 105 mL.min(-1).1.73 m(-2). Mendelian randomization analyses for CHD showed an association among participants with eGFR 105 mL.min(-1).1.73 m(-2). Results were not materially different after adjustment for factors associated with the eGFR genetic risk score, such as lipoprotein(a), triglycerides, hemoglobin Alc, and blood pressure. Mendelian randomization results for stroke were nonsignificant but broadly similar to those for CHD. CONCLUSIONS: In people without manifest cardiovascular disease or diabetes, mild-to-moderate kidney dysfunction is causally related to risk of CHD, highlighting the potential value of preventive approaches that preserve and modulate kidney function