Droplet-based microfluidic screening and sorting of microalgal populations for strain engineering applications

The application of microfluidic technologies to microalgal research is particularly appealing since these approaches allow the precise control of the extracellular environment and offer a high-throughput approach to studying dynamic cellular processes. To expand the portfolio of applications, here we present a droplet-based microfluidic method for analysis and screening of Phaeodactylum tricornutum and Nannochloropsis gaditana, which can be integrated into a genetic transformation workflow. Following encapsulation of single cells in picolitre-sized droplets, fluorescence signals arising from each cell can be used to assess its phenotypic state. In this work, the chlorophyll fluorescence intensity of each cell was quantified and used to identify populations of P. tricornutum cells grown in different light conditions. Further, individual P. tricornutum or N. gaditana cells engineered to express green fluorescent protein were distinguished and sorted from wild-type cells. This has been exploited as a rapid screen for transformed cells within a population, bypassing a major bottleneck in algal transformation workflows and offering an alternative strategy for the identification of genetically modified strains

Antiviral CD4+ memory T cells are IL-15 dependent

Survival and intermittent proliferation of memory CD4+ and CD8+ T cells appear to be controlled by different homeostatic mechanisms. In particular, contact with interleukin (IL)-15 has a decisive influence on memory CD8+ cells, but not memory CD4+ cells. Past studies of memory CD4+ cells have relied heavily on the use of naturally occurring memory phenotype (MP) cells as a surrogate for antigen (Ag)-specific memory cells. However, we show here that MP CD4+ cells contain a prominent subset of rapidly proliferating major histocompatibility complex (MHC) II–dependent cells. In contrast, Ag-specific memory CD4 cells have a slow turnover rate and are MHC II independent. In irradiated hosts, these latter cells ignore IL-15 and expand in response to the elevated levels of IL-7 in the lymphopenic hosts. In contrast, in normal nonlymphopenic hosts where IL-7 levels are low, memory CD4 cells are heavily dependent on IL-15. Significantly, memory CD4+ responsiveness to endogenous IL-15 reflects marked competition from other cells, especially CD8+ and natural killer cells, and increases considerably after removal of these cells. Therefore, under normal physiological conditions, homeostasis of CD8+ and CD4+ memory cells is quite similar and involves IL-15 and IL-7

The Distance Scale of Planetary Nebulae

By collecting distances from the literature, a set of 73 planetary nebulae with mean distances of high accuracy is derived. This sample is used for recalibration of the mass-radius relationship, used by many statistical distance methods. An attempt to correct for a statistical peculiarity, where errors in the distances influences the mass--radius relationship by increasing its slope, has been made for the first time. Distances to PNe in the Galactic Bulge, derived by this new method as well as other statistical methods from the last decade, are then used for the evaluation of these methods as distance indicators. In order of achieving a Bulge sample that is free from outliers we derive new criteria for Bulge membership. These criteria are much more stringent than those used hitherto, in the sense that they also discriminate against background objects. By splitting our Bulge sample in two, one with optically thick (small) PNe and one with optically thin (large) PNe, we find that our calibration is of higher accuracy than most other calibrations. Differences between the two subsamples, we believe, are due to the incompleteness of the Bulge sample, as well as the dominance of optical diameters in the ``thin'' sample and radio diameters in the ``thick'' sample. Our final conclusion is that statistical methods give distances that are at least as accurate as the ones obtained from many individual methods. Also, the `long' distance scale of Galactic PNe is confirmed.Comment: 15 pages, 9 figures, accepted for publication in A&

The Synthesis Telescope at the Dominion Radio Astrophysical Observatory

We describe an aperture synthesis radio telescope optimized for studies of the Galactic interstellar medium (ISM), providing the ability to image extended structures with high angular resolution over wide fields. The telescope produces images of atomic hydrogen emission using the 21-cm HI spectral line, and, simultaneously, continuum emission in two bands centred at 1420 MHz and 408 MHz, including linearly polarized emission at 1420 MHz, with synthesized beams of 1 degree and 3.4 degrees at the respective frequencies.Comment: Accepted for publication by Astronomy and Astrophysics, Supplement Serie

Health and Well-Being of International University Students, and Comparison with Domestic Students, in Tasmania, Australia

International students comprise an increasingly larger proportion of higher education students globally. Empirical evidence about the health and well-being of these students is, however, limited. We sought to examine the health and well-being of international students, primarily from Asian countries, attending the University of Tasmania, Australia, using domestic students as a comparison group. Ethics approval was given to invite (via email) all currently enrolled students to participate in the study by completing a pilot-tested, online survey. The survey was completed by 382 international students (response rate = 8.9%) and 1013 domestic students (9.2%). Independent samples t-tests, analysis of variance (ANOVA) and chi-square tests were used for bivariate comparisons between international and domestic students, and between subgroups of international students. Regression models were used to examine the associations between student status (international vs. domestic) and health outcomes, controlling for demographic and enrolment variables. International students, particularly male students, were found to be at increased risk of several adverse health outcomes while also being less likely to seek help for mental health and related problems. The findings indicate the need for accessible, targeted, culturally-sensitive health promotion and early intervention programs

Successful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes

The microbial parasite Blastocystis colonizes the large intestines of numerous animal species and increasing evidence has linked Blastocystis infection to enteric diseases with signs and symptoms including abdominal pain, constipation, diarrhea, nausea, vomiting, and flatulence. It has also recently been reported to be an important member of the host intestinal microbiota. Despite significant advances in our understanding of Blastocystis cell biology and host-parasite interactions, a genetic modification tool is absent. In this study, we successfully established a robust gene delivery protocol for Blastocystis subtype 7 (ST7) and ectopic protein expression was further tested using a high sensitivity nano-luciferase (Nluc) reporter system, with promoter regions from several genes. Among them, a strong promoter encompassing a region upstream of the legumain 5? UTR was identified. Using this promoter combined with the legumain 3? UTR, which contains a conserved, precise polyadenylation signal, a robust transient transfection technique was established for the first time in Blastocystis. This system was validated by ectopic expression of proteins harbouring specific localization signals. The establishment of a robust, reproducible gene modification system for Blastocystis is a significant advance for Blastocystis research both in vitro and in vivo. This technique will spearhead further research to understand the parasite’s biology, its role in health and disease, along with novel ways to combat the parasite

The aged lymphoid tissue environment fails to support naive T cell homeostasis.

Aging is associated with a gradual loss of naive T cells and a reciprocal increase in the proportion of memory T cells. While reduced thymic output is important, age-dependent changes in factors supporting naive T cells homeostasis may also be involved. Indeed, we noted a dramatic decrease in the ability of aged mice to support survival and homeostatic proliferation of naive T cells. The defect was not due to a reduction in IL-7 expression, but from a combination of changes in the secondary lymphoid environment that impaired naive T cell entry and access to key survival factors. We observed an age-related shift in the expression of homing chemokines and structural deterioration of the stromal network in T cell zones. Treatment with IL-7/mAb complexes can restore naive T cell homeostatic proliferation in aged mice. Our data suggests that homeostatic mechanisms that support the naive T cell pool deteriorate with age.11128Ysciescopu

Experimental and computational study of trace element distribution between orthopyroxene and anhydrous silicate melts: substitution mechanisms and the effect of iron

Although orthopyroxene (Opx) is present during a wide range of magmatic differentiation processes in the terrestrial and lunar mantle, its effect on melt trace element contents is not well quantified. We present results of a combined experimental and computational study of trace element partitioning between Opx and anhydrous silicate melts. Experiments were performed in air at atmospheric pressure and temperatures ranging from 1,326 to 1,420°C in the system CaO-MgO-A

The \u3cem\u3eChlamydomonas\u3c/em\u3e Genome Reveals the Evolution of Key Animal and Plant Functions

Chlamydomonas reinhardtii is a unicellular green alga whose lineage diverged from land plants over 1 billion years ago. It is a model system for studying chloroplast-based photosynthesis, as well as the structure, assembly, and function of eukaryotic flagella (cilia), which were inherited from the common ancestor of plants and animals, but lost in land plants. We sequenced the ∼120-megabase nuclear genome of Chlamydomonas and performed comparative phylogenomic analyses, identifying genes encoding uncharacterized proteins that are likely associated with the function and biogenesis of chloroplasts or eukaryotic flagella. Analyses of the Chlamydomonas genome advance our understanding of the ancestral eukaryotic cell, reveal previously unknown genes associated with photosynthetic and flagellar functions, and establish links between ciliopathy and the composition and function of flagella

Ablation of the Pro-Apoptotic Protein Bax Protects Mice from Glucocorticoid-Induced Bone Growth Impairment

Dexamethasone (Dexa) is a widely used glucocorticoid to treat inflammatory diseases; however, a multitude of undesired effects have been reported to arise from this treatment including osteoporosis, obesity, and in children decreased longitudinal bone growth. We and others have previously shown that glucocorticoids induce apoptosis in growth plate chondrocytes. Here, we hypothesized that Bax, a pro-apoptotic member of the Bcl-2 family, plays a key role in Dexa-induced chondrocyte apoptosis and bone growth impairment. Indeed, experiments in the human HCS-2/8 chondrocytic cell line demonstrated that silencing of Bax expression using small-interfering (si) RNA efficiently blocked Dexa-induced apoptosis. Furthermore, ablation of Bax in female mice protected against Dexa-induced bone growth impairment. Finally, Bax activation by Dexa was confirmed in human growth plate cartilage specimens cultured ex vivo. Our findings could therefore open the door for new therapeutic approaches to prevent glucocorticoid-induced bone growth impairment through specific targeting of Bax