Absence of nucleolar dominance in mouse-human heterokaryons

Autoradiographic analysis of [³H]uridine incorporation 48 h after polyethylene glycol-mediated cell fusion indicates that nucleolar RNA synthesis persists in both human and mouse nuclei in interspecific heterokaryons. The absence of nucleolar dominance in heterokaryons has been confirmed by zinc-dithizone nucleolus-specific staining, and is true even when there are considerably more nuclei of one species than of the other in the heterokaryon. Studies of actinomycin D-induced nucleolar segregation indicate that the zinc-binding proteins responsible for zinc-dithizone staining are located in a different nucleolar component than the protein responsible for silver staining.     

Experimental Adaptation of Rotaviruses to Tumor Cell Lines

A number of viruses show a naturally extended tropism for tumor cells whereas other viruses have been genetically modified or adapted to infect tumor cells. Oncolytic viruses have become a promising tool for treating some cancers by inducing cell lysis or immune response to tumor cells. In the present work, rotavirus strains TRF-41 (G5) (porcine), RRV (G3) (simian), UK (G6-P5) (bovine), Ym (G11-P9) (porcine), ECwt (murine), Wa (G1-P8), Wi61 (G9) and M69 (G8) (human), and five wild-type human rotavirus isolates were passaged multiple times in different human tumor cell lines and then combined in five different ways before additional multiple passages in tumor cell lines. Cell death caused by the tumor cell-adapted isolates was characterized using Hoechst, propidium iodide, 7-AAD, Annexin V, TUNEL, and anti-poly-(ADP ribose) polymerase (PARP) and -phospho-histone H2A.X antibodies. Multiple passages of the combined rotaviruses in tumor cell lines led to a successful infection of these cells, suggesting a gain-of-function by the acquisition of greater infectious capacity as compared with that of the parental rotaviruses. The electropherotype profiles suggest that unique tumor cell-adapted isolates were derived from reassortment of parental rotaviruses. Infection produced by such rotavirus isolates induced chromatin modifications compatible with apoptotic cell death.     

Turning regenerative technologies into treatment to repair myocardial injuries

Regenerative therapies including stem cell treatments hold promise to allow curing patients affected by severe cardiac muscle diseases. However, the clinical efficacy of stem cell therapy remains elusive, so far. The two key roadblocks that still need to be overcome are the poor cell engraftment into the injured myocardium and the limited knowledge of the ideal mixture of bioactive factors to be locally delivered for restoring heart function. Thus, therapeutic strategies for cardiac repair are directed to increase the retention and functional integration of transplanted cells in the damaged myocardium or to enhance the endogenous repair mechanisms through cell-free therapies. In this context, biomaterial-based technologies and tissue engineering approaches have the potential to dramatically impact cardiac translational medicine. This review intends to offer some consideration on the cell-based and cell-free cardiac therapies, their limitations and the possible future developments.     

Nest box revealed habitat preferences of arboreal mammals in box‐ironbark forest

Habitat preferences need to be understood if species are to be adequately managed or conserved. Habitat preferences are presumed to reflect requirements for food, shelter and breeding, as well as interactions with predators and competitors. However, one or more of these requirements may dominate. Tree‐cavity‐dependent wildlife species are one example where shelter or breeding site requirements may dominate. We installed 120 nest boxes across 40 sites to target the vulnerable Brush‐tailed Phascogale (Phascogale tapoatafa) and the non‐threatened Sugar Glider (Petaurus breviceps). The provision of shelter sites where few of quality are available may enable better resolution of habitat preferences. Over three years, we observed the Brush‐tailed Phascogale at 17 sites, whereas the Sugar Glider was observed at 39 sites. We tested four broad hypotheses (H1–H4) relating to habitat that may influence occupancy by these species. There was no influence of hollow (cavity) abundance (H1) on either species suggesting our nest boxes had satisfied their shelter requirements. There was no influence of habitat structure (canopy and tree proximity) (H2) immediately around the nest box trees. We found no influence of distance to the forest edge (H3). Variables at and away from the nest box site that appear to reflect foraging substrates (H4) were influential on the Brush‐tailed Phascogale. Sugar Glider occupancy was only influenced by a single variable at the nest box site. The lack of influence of any other variables is consistent with the very high occupancy observed, suggesting most of the forest habitat is suitable when shelter sites are available. We found no evidence that the Sugar Glider reduced site use by the Brush‐tailed Phascogale.     

Reduction of medium consumption in perfusion mammalian cell cultures using a perfusion rate equivalent concentrated nutrient feed

Media preparation for perfusion cell culture processes contributes significantly to operational costs and the footprint of continuous operations for therapeutic protein manufacturing. In this study, definitions are given for the use of a perfusion equivalent nutrient feed stream which, when used in combination with basal perfusion medium, supplements the culture with targeted compounds and increases the medium depth. Definitions to compare medium and feed depth are given in this article. Using a concentrated nutrient feed, a 1.8-fold medium consumption (MC) decrease and a 1.67-fold increase in volumetric productivity (PR) were achieved compared to the initial condition. Later, this strategy was used to push cell densities above 100 × 10⁶ cells/ml while using a perfusion rate below 2 RV/day. In this example, MC was also decreased 1.8-fold compared to the initial condition, but due to the higher cell density, PR was increased 3.1-fold and to an average PR value of 1.36 g L⁻¹ day⁻¹ during a short stable phase, and versus 0.46 g L⁻¹ day⁻¹ in the initial condition. Overall, the performance improvements were aligned with the given definitions. This multiple feeding strategy can be applied to gain some flexibility during process development and also in a manufacturing set-up to enable better control on nutrient addition.     

Characterization of Indoor Environmental Quality in Primary Schools in Maia: A Portuguese Case Study

Scientific evidence associates indoor environment pollutants with respiratory effects (asthma and allergies), and children constitute one of most sensitive groups. Indoor air quality (IAQ) in schools may indeed be a significant health factor for children, with effects on school attendance and performance. Our aim was to characterize IAQ of classrooms in Maia County (north of Portugal) for which there was no information available. The study was conducted in 21 of the 40 primary schools, selected by stratified random sampling. Depending on the dimension, one or two classrooms were tested at each school. Walkthrough surveys of school grounds, buildings, and individual classrooms were done. Continuous measurements were taken of temperature, relative humidity, airborne respirable particles, total volatile organic compounds, and carbon dioxide, whereas bioaerosols were counted on Plate Count Agar during regular school activities. The indoor arithmetic mean for PM₁₀, CO₂, TCOV, and bioaerosol concentrations were 0.14 mg/m³, 999 ppm, 0.41 mg/m³, and 4140 UCF/m³, respectively. The values of PM₁₀ and CO₂ were close to their acceptable maximum values, but bioaerosols were much higher. This study supports previous studies conducted in school environments and emphasizes the need for proactive indoor air quality audits in school buildings.     

Using Information Interaction to Discover Epistatic Effects in Complex Diseases

It is widely agreed that complex diseases are typically caused by the joint effects of multiple instead of a single genetic variation. These genetic variations may show stronger effects when considered together than when considered individually, a phenomenon known as epistasis or multilocus interaction. In this work, we explore the applicability of information interaction to discover pairwise epistatic effects related with complex diseases. We start by showing that traditional approaches such as classification methods or greedy feature selection methods (such as the Fleuret method) do not perform well on this problem. We then compare our information interaction method with BEAM and SNPHarvester in artificial datasets simulating epistatic interactions and show that our method is more powerful to detect pairwise epistatic interactions than its competitors. We show results of the application of information interaction method to the WTCCC breast cancer dataset. Our results are validated using permutation tests. We were able to find 89 statistically significant pairwise interactions with a p-value lower than . Even though many recent algorithms have been designed to find epistasis with low marginals, we observed that all (except one) of the SNPs involved in statistically significant interactions have moderate or high marginals. We also report that the interactions found in this work were not present in gene-gene interaction network STRING.