Production and Utility of Extracellular Vesicles with 3D Culture Methods

In recent years, extracellular vesicles (EVs) have emerged as promising biomarkers, cell-free therapeutic agents, and drug delivery carriers. Despite their great clinical potential, poor yield and unscalable production of EVs remain significant challenges. When using 3D culture methods, such as scaffolds and bioreactors, large numbers of cells can be expanded and the cell environment can be manipulated to control the cell phenotype. This has been employed to successfully increase the production of EVs as well as to enhance their therapeutic effects. The physiological relevance of 3D cultures, such as spheroids, has also provided a strategy for understanding the role of EVs in the pathogenesis of several diseases and to evaluate their role as tools to deliver drugs. Additionally, 3D culture methods can encapsulate EVs to achieve more sustained therapeutic effects as well as prevent premature clearance of EVs to enable more localised delivery and concentrated exosome dosage. This review highlights the opportunities and drawbacks of different 3D culture methods and their use in EV research

Essential Roles of Drosophila Inner Centromere Protein (Incenp) and Aurora B in Histone H3 Phosphorylation, Metaphase Chromosome Alignment, Kinetochore Disjunction, and Chromosome Segregation

We have performed a biochemical and double-stranded RNA-mediated interference (RNAi) analysis of the role of two chromosomal passenger proteins, inner centromere protein (INCENP) and aurora B kinase, in cultured cells of Drosophila melanogaster. INCENP and aurora B function is tightly interlinked. The two proteins bind to each other in vitro, and DmINCENP is required for DmAurora B to localize properly in mitosis and function as a histone H3 kinase. DmAurora B is required for DmINCENP accumulation at centromeres and transfer to the spindle at anaphase. RNAi for either protein dramatically inhibited the ability of cells to achieve a normal metaphase chromosome alignment. Cells were not blocked in mitosis, however, and entered an aberrant anaphase characterized by defects in sister kinetochore disjunction and the presence of large amounts of amorphous lagging chromatin. Anaphase A chromosome movement appeared to be normal, however cytokinesis often failed. DmINCENP and DmAurora B are not required for the correct localization of the kinesin-like protein Pavarotti (ZEN-4/CHO1/MKLP1) to the midbody at telophase. These experiments reveal that INCENP is required for aurora B kinase function and confirm that the chromosomal passengers have essential roles in mitosis

A state-of-the-art review on birds as indicators of biodiversity : Advances, challenges, and future directions

The current loss of biodiversity has been broadly acknowledged as the main cause of ecosystem change. To halt this trend, several international agreements have been made, and various biodiversity metrics have been developed to evaluate whether the targets of these agreements are being met. The process of developing good indicators is not trivial. Indicators should be able to synthesize and communicate our current knowledge, but they also need to meet both scientific and practical criteria. Since it would not be practical to monitor all species, indicators are mainly built on the monitoring of some well-known taxa, such as birds. Here we systematically review the wide spectrum of bird biodiversity indicators (hereafter indicators) available to: i) evaluate recent methodological advances; ii) identify current knowledge gaps jeopardizing indicator interpretation and use in guiding decision-making; and iii) examine challenges in their applicability across different spatial and temporal contexts. We pay particular attention to indicator characteristics such as site and species selection, spatial, seasonal and habitat coverage, and statistical issues in developing indicators and tools to tackle them, to provide specific recommendations for the future construction of indicators. Several methodological advances have recently been made to enhance the process of indicator development, including multiple ways to select sites and species to increase their robustness. However, we found that there are strong spatial, seasonal and habitat biases among the selected indicators. Most of them are from Europe, using mainly census data from the breeding season and typically covering farmland and forest habitats. The major challenges that we detected in their applicability were related to the modelling of the statistical uncertainty associated to the indicator. We recommend the use of quantitative methods in site and species selection procedures whenever possible. Current indicators should be expanded to areas outside Europe and to less studied habitats and should not neglect monitoring work outside the breeding season. Time-series analyses studying temporal trends and using multi-species data should in general account for temporal autocorrelation as well as for phylogenetic correlation. Multi-species hierarchical models are a good alternative for analysing and constructing indicators, but they need to include annual random effects allowing for unexplained annual variation in the average status of the community, i.e. the indicator target. Despite methodological and context-specific differences in the indicators reviewed, most of them seem to highlight the urgent need of devising strategic climate and conservation policies to improve the status and trends of biodiversity.Peer reviewe

Optimal 57Co Flood Source Activity and Acquisition Time for Lymphoscintigraphy Localization Images

Martha V. Mar, Renee L. Dickinson, William D. Erwin, Richard E. Wendt. Optimal 57Co Flood Source Activity and Acquisition Time for Lymphoscintigraphy Localization Images. Journal of Nuclear Medicine Technology Jun 2008, 36 (2) 82-87; DOI: 10.2967/jnmt.107.047043https://openworks.mdanderson.org/mdacc_imgphys_pubs/1008/thumbnail.jp

Joyce’s heirs. Joyce’s imprint on recent global literatures

166 p.The figure of James Joyce is intangible, an almost all-encompassing figure whose height and breadth bypasses countries, continents and even time constraints and limitations. Many writers have confessed to their being indebted to his works and readings, as have many scholars over the years. However, not everything said about Joyce has always been praising; his contemporary D.H. Lawrence is known to have criticised him on the basis of his Biblical references or his journalistic-indebted narrative, which he defined as “old and hard-worked staleness, masquerading as the all-new”. Many more writers have, however, found inspiration in Joyce’s narratives and stories than not. Admirers and detractors aside, it is clear that Joyce’s figure is larger than life. Almost as a reversed parallelism to D.H. Lawrence’s criticism, Joyce can be pronounced a figure of almost Biblical proportions. One may like him or despise him; however, no one is left indifferent by it. This lofty, academic assertion has its more mundane mirror image in the widespread myth that all Dubliners, upon entering a conversation of literary dimensions, will firmly state their own opinions on the Dubliner’s work, even discuss some of his passages, only to later acknowledge (perhaps in the intimacy of one of those public houses Joyce himself so well depicted) that they have not read the book at all. This, rather than taking away literary value form Joyce’s work, proves how all-encompassing and ever-arching his work can be. Discussed alike by highranking academics and Dublin taxi drivers, Joyce’s oeuvre is an ever-continuing metaphor of life and its essence at the core of the city which he left but always inhabited

Dysregulated cell homeostasis and miRNAs in human iPSC-derived cardiomyocytes from a propionic acidemia patient with cardiomyopathy

Propionic acidemia (PA) disorder shows major involvement of the heart, among other alterations. A significant number of PA patients develop cardiac complications, and available evidence suggests that this cardiac dysfunction is driven mainly by the accumulation of toxic metabolites. To contribute to the elucidation of the mechanistic basis underlying this dysfunction, we have successfully generated cardiomyocytes through the differentiation of induced pluripotent stem cells (iPSCs) from a PCCB patient and its isogenic control. In this human cellular model, we aimed to examine microRNAs (miRNAs) profiles and analyze several cellular pathways to determine miRNAs activity patterns associated with PA cardiac phenotypes. We have identified a series of upregulated cardiac-enriched miRNAs and alterations in some of their regulated signaling pathways, including an increase in the expression of cardiac damage markers and cardiac channels, an increase in oxidative stress, a decrease in mitochondrial respiration and autophagy; and lipid accumulation. Our findings indicate that miRNA activity patterns from PA iPSC-derived cardiomyocytes are biologically informative and advance the understanding of the molecular mechanisms of this rare disease, providing a basis for identifying new therapeutic targets for intervention strategie

Drosophila Polo Kinase Is Required for Cytokinesis

A number of lines of evidence point to a predominance of cytokinesis defects in spermatogenesis in hypomorphic alleles of the Drosophila polo gene. In the pre-meiotic mitoses, cytokinesis defects result in cysts of primary spermatocytes with reduced numbers of cells that can contain multiple centrosomes. These are connected by a correspondingly reduced number of ring canals, structures formed by the stabilization of the cleavage furrow. The earliest defects during the meiotic divisions are a failure to form the correct mid-zone and mid-body structures at telophase. This is accompanied by a failure to correctly localize the Pavarotti kinesin- like protein that functions in cytokinesis, and of the septin Peanut and of actin to be incorporated into a contractile ring. In spite of these defects, cyclin B is degraded and the cells exit M phase. The resulting spermatids are frequently binuclear or tetranuclear, in which case they develop either two or four axonemes, respectively. A significant proportion of spermatids in which cytokinesis has failed may also show the segregation defects previously ascribed to polo1 mutants. We discuss these findings in respect to conserved functions for the Polo-like kinases in regulating progression through M phase, including the earliest events of cytokinesis

Designs and Optical Tests of Thermal Links for an Optical Refrigerator

Dielectric mirror leakage at large angles of incidence limits the effectiveness of solid state optical refrigerators due to reheating caused by photon absorption in an attached load. In this paper, we present several thermally conductive link solutions to greatly reduce the net photon absorption. The Los Alamos Solid State Optical Refrigerator (LASSOR) has demonstrated cooling of a Yb/sup 3+/ doped ZBLANP glass to 208 K. We have designed optically isolating thermal link geometries capable of extending cooling to a typical heat load with minimal absorptive reheating, and we have tested the optical performance of these designs. A surrogate source operating at 625 nm was used to mimic the angular distribution of light from the LASSOR cooling element. While total link performance is dependent on additional factors, we have found that the best thermal link reduced the net transmission of photons to 0.04%, which includes the trapping mirrors 8.1% transmission. Our measurements of the optical performance of the various link geometries are supported by computer simulations of the designs using Code V, a commercially available optical modeling software package

Instanton size distribution in O(3)

We present calculations of the size distribution of instantons in the 2d O(3) non-linear sigma-model, and briefly discuss the effects cooling has upon the configurations and the topological objects. (This preprint is also available via anonymous ftp to suna.amtp.liv.ac.uk in /pub/pss/ as instdist.uue.)Comment: 17 pages, LaTeX, needs cite.sty (appended), with appended uuencoded compressed tarfile of PostScript figures, Liverpool preprint LTH-33

Computer Modeling and Analysis of Thermal Link Performance for an Optical Refrigerator

We have used the thermal modeling tool in COMSOL Multiphysics to investigate factors that affect the thermal performance of the optical refrigerator. Assuming an ideal cooling element and a non-absorptive dielectric trapping mirror, the three dominant heating factors are blackbody radiation from the surrounding environment, conductive heat transfer through mechanical supports, and the absorption of fluorescent photons transmitted through the thermal link. Laboratory experimentation coupled with computer modeling using Code V optical software have resulted in link designs capable of reducing the transmission to 0.04% of the fluoresced photons emitted toward the thermal link. The ideal thermal link will have minimal surface area, provide complete optical isolation for the load, and possess high thermal conductivity. Modeling results imply that a 1 cm/sup 3/ load can be chilled to 102 K with currently available cooling efficiencies using a 100 W pump laser on a YB:ZBLANP system, and using an ideal link that has minimal surface area and no optical transmission. We review the simulated steady-state cooling temperatures reached by the heat load for several link designs and system configurations as a comparative measure of how well particular configurations perform