Puromycin-based vectors promote a ROS-dependent recruitment of PML to nuclear inclusions enriched with HSP70 and Proteasomes

<p>Abstract</p> <p>Background</p> <p>Promyelocytic Leukemia (PML) protein can interact with a multitude of cellular factors and has been implicated in the regulation of various processes, including protein sequestration, cell cycle regulation and DNA damage responses. Previous studies reported that misfolded proteins or proteins containing polyglutamine tracts form aggregates with PML, chaperones, and components of the proteasome, supporting a role for PML in misfolded protein degradation.</p> <p>Results</p> <p>In the current study, we have identified a reactive oxygen species (ROS) dependent aggregation of PML, small ubiquitin-like modifier 1 (SUMO-1), heat shock protein 70 (HSP70) and 20S proteasomes in human cell lines that have been transiently transfected with vectors expressing the puromycin resistance gene, puromycin n-acetyl transferase (pac). Immunofluorescent studies demonstrated that PML, SUMO-1, HSP70 and 20S proteasomes aggregated to form nuclear inclusions in multiple cell lines transfected with vectors expressing puromycin (puro) resistance in regions distinct from nucleoli. This effect does not occur in cells transfected with identical vectors expressing other antibiotic resistance genes or with vectors from which the pac sequence has been deleted. Furthermore, ROS scavengers were shown to ablate the effect of puro vectors on protein aggregation in transfected cells demonstrating a dependency of this effect on the redox state of transfected cells.</p> <p>Conclusion</p> <p>Taken together we propose that puromycin vectors may elicit an unexpected misfolded protein response, associated with the formation of nuclear aggresome like structures in human cell lines. This effect has broad implications for cellular behavior and experimental design.</p

Facilitating smart HACCP strategies with Process Analytical Technology

The critical feature to any food management system, be it safety or quality focused, is timely and accurate product and process data. The HACCP framework is one of a systematic preventive approach that addresses hazards as a means of prevention rather than finished product inspection. The PAT initiative is a framework which also aims to change product assurance from end-product testing to preventive control through timely measurements of critical product and process parameters. The continued emergence of sensitive yet affordable sensors capable of direct measurement of biological, chemical and physical risks associated with foods during production, processing or over their shelf-life undoubtedly will support a higher temporal and spatial profiling of foods

Elemental analysis of fish feed by laser-induced breakdown spectroscopy

In this study, the potential of laser-induced breakdown spectroscopy (LIBS) as an efficient multi-elemental quantification tool for fish feed is determined. A particular focus of this paper is total chromium, an essential element that has the potential to be toxic and carcinogenic. In total six elements, four macro-elements (Ca, Fe, K and Mg) and two micro-elements (Cr and Rb), were modelled using LIBS spectra of aquafeed samples. Reference analysis was conducted via inductively coupled plasma mass spectrometry (ICP-MS) and showed good agreement with LIBS predictions. These results provide evidence that LIBS has the potential to be utilized in the field as a real-time screening tool for establishing the elemental composition of a range of fish feeds

VLBA Imaging of the OH Maser in IIIZw35

We present a parsec-scale image of the OH maser in the nucleus of the active galaxy IIIZw35, made using the Very Long Baseline Array at a wavelength of 18 cm. We detected two distinct components, with a projected separation of 50 pc (for D=110 Mpc) and a separation in Doppler velocity of 70 km/s, which contain 50% of the total maser flux. Velocity gradients within these components could indicate rotation of clouds with binding mass densities of ~7000 solar masses per cubic parsec, or total masses of more than 500,000 solar masses. Emission in the 1665-MHz OH line is roughly coincident in position with that in the 1667-MHz line, although the lines peak at different Doppler velocities. We detected no 18 cm continuum emission; our upper limit implies a peak apparent optical depth greater than 3.4, assuming the maser is an unsaturated amplifier of continuum radiation.Comment: 10 pages, 3 figure

Low fitness partially explains resting metabolic rate differences between African American and white women

Background High levels of obesity among African American women have been hypothesized to be partially resultant from a lower resting metabolic rate compared with white women. The aim of the current study was to determine if differences in cardiorespiratory fitness and moderate-to-vigorous physical activity are associated with differences in resting metabolic rate among free-living young adult African American women and white women. Methods Participants were 179 women (white women n = 141, African American women n = 38, mean age = 27.7 years). Resting metabolic rate was measured using indirect calorimetry, body composition using dual energy x-ray absorptiometry, cardiorespiratory fitness via maximal treadmill test, and moderate-to-vigorous physical activity using an activity monitor. Results African American women had higher body mass index, fat mass, and fat-free mass compared with white women but lower levels of cardiorespiratory fitness. No differences were observed between African American and white women in resting metabolic rate when expressed as kcal/day (1390.8 ± 197.5 vs 1375.7 ± 173.6 kcal/day, P =.64), but African American women had a lower resting metabolic rate when expressed relative to body weight (2.56 ± 0.30 vs 2.95 ± 0.33 mL/kg/min,

Impacts of the invasive round goby (Neogobius melanostomus) on benthic invertebrate fauna: a case study from the Baltic Sea

The round goby (Neogobius melanostomus) was first observed in the Baltic Sea in 1990 and has since displayed substantial secondary dispersal, establishing numerous dense populations where they may outcompete native fish and negatively impact prey species. There have been multiple round goby diet studies from both the Baltic Sea and the North American Great Lakes where they are similarly invasive. However, studies that quantify their effects on recipient ecosystems and, specifically, their impacts on the benthic invertebrate macrofauna are rare, particularly from European waters. In this study, we conducted the first before-after study of the potential effects of round goby on benthic invertebrate macrofauna taxa in marine-brackish habitats in Europe, focusing of two sites in the Western Baltic Sea, Denmark. Results were in line with those from the Great Lakes, indicating negative impacts on specific molluscan taxa (e.g. Cardiidae bivalves and Neritidae gastropods, which both showed a fall in detected densities of approximately 98% within the Guldborgsund Strait). In contrast, many other groups appeared to be largely unaffected or even show positive trends following invasion. Round goby gut content data were available at one of our study sites from the period immediately after the invasion. These data confirmed that round goby had in fact been preying on the subset of taxa displaying negative trends

Identification of a Novel Keyhole Phenotype in Double-Disk Diffusion Assays of Clindamycin-Resistant Erythromycin-Sensitive Strains of Streptococcus agalactiae

Our objective was to characterize 46 unique, erythromycin-sensitive, and clindamycin-resistant Streptococcus agalactiae strains from S. Korea that displayed a novel phenotype in double-disk diffusion assay. We used polymerase chain reaction to determine presence of erythromycin and clindamycin resistance genes, disc diffusion assays to determine resistance phenotype, and microbroth dilution to determine minimal inhibitory concentration. We detected a novel phenotype in the double-disk diffusion assay for inducible resistance among 46 S. agalactiae strains that were both erythromycin sensitive and clindamycin resistant. Thirty-two strains with the novel phenotype tested positive for erm(B) by DNA-DNA hybridization; sequencing of the erm(B) gene revealed mutations in the ribosomal binding site region in the erm(B) open reading frame, which is consistent with a lack of erythromycin resistance phenotype. Although identified from patients at multiple hospitals, genotyping suggested that the strains are closely related. The new phenotype shows increased sensitivity to clindamycin in the presence of erythromycin.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/90482/1/mdr-2E2010-2E0040.pd

The emerging field of venom-microbiomics for exploring venom as a microenvironment, and the corresponding Initiative for Venom Associated Microbes and Parasites (iVAMP)

Venom is a known source of novel antimicrobial natural products. The substantial, increasing number of these discoveries have unintentionally culminated in the misconception that venom and venom-producing glands are largely sterile environments. Culture-dependent and -independent studies on the microbial communities in venom microenvironments reveal the presence of archaea, algae, bacteria, endoparasites, fungi, protozoa, and viruses. Venom-centric microbiome studies are relatively sparse to date and the adaptive advantages that venom-associated microbes might offer to their hosts, or that hosts might provide to venom-associated microbes, remain unknown. We highlight the potential for the discovery of venom-microbiomes within the adaptive landscape of venom systems. The considerable number of known, convergently evolved venomous animals juxtaposed with the comparatively few studies to identify microbial communities in venom provides new possibilities for both biodiversity and therapeutic discoveries. We present an evidence-based argument for integrating microbiology as part of venomics to which we refer to as venom-microbiomics. We also introduce iVAMP, the Initiative for Venom Associated Microbes and Parasites (https://ivamp-consortium.github.io/), as a growing consortium for interested parties to contribute and collaborate within this subdiscipline. Our consortium seeks to support diversity, inclusion and scientific collaboration among all researchers interested in this subdiscipline

Infrastructural Speculations: Tactics for Designing and Interrogating Lifeworlds

This paper introduces “infrastructural speculations,” an orientation toward speculative design that considers the complex and long-lived relationships of technologies with broader systems, beyond moments of immediate invention and design. As modes of speculation are increasingly used to interrogate questions of broad societal concern, it is pertinent to develop an orientation that foregrounds the “lifeworld” of artifacts—the social, perceptual, and political environment in which they exist. While speculative designs often imply a lifeworld, infrastructural speculations place lifeworlds at the center of design concern, calling attention to the cultural, regulatory, environmental, and repair conditions that enable and surround particular future visions. By articulating connections and affinities between speculative design and infrastructure studies research, we contribute a set of design tactics for producing infrastructural speculations. These tactics help design researchers interrogate the complex and ongoing entanglements among technologies, institutions, practices, and systems of power when gauging the stakes of alternate lifeworlds

The Scattering and Intrinsic Structure of Sagittarius A* at Radio Wavelengths

Radio images of the Galactic Center supermassive black hole, Sagittarius A* (Sgr A*), are dominated by interstellar scattering. Previous studies of Sgr A* have adopted an anisotropic Gaussian model for both the intrinsic source and the scattering, and they have extrapolated the scattering using a purely $\lambda^2$ scaling to estimate intrinsic properties. However, physically motivated source and scattering models break all three of these assumptions. They also predict that refractive scattering effects will be significant, which have been ignored in standard model fitting procedures. We analyze radio observations of Sgr A* using a physically motivated scattering model, and we develop a prescription to incorporate refractive scattering uncertainties when model fitting. We show that an anisotropic Gaussian scattering kernel is an excellent approximation for Sgr A* at wavelengths longer than 1cm, with an angular size of $(1.380 \pm 0.013) \lambda_{\rm cm}^2\,{\rm mas}$ along the major axis, $(0.703 \pm 0.013) \lambda_{\rm cm}^2\,{\rm mas}$ along the minor axis, and a position angle of $81.9^\circ \pm 0.2^\circ$. We estimate that the turbulent dissipation scale is at least $600\,{\rm km}$, with tentative support for $r_{\rm in} = 800 \pm 200\,{\rm km}$, suggesting that the ion Larmor radius defines the dissipation scale. We find that the power-law index for density fluctuations in the scattering material is $\beta < 3.47$, shallower than expected for a Kolmogorov spectrum ($\beta=11/3$), and we estimate $\beta = 3.38^{+0.08}_{-0.04}$ in the case of $r_{\rm in} = 800\,{\rm km}$. We find that the intrinsic structure of Sgr A* is nearly isotropic over wavelengths from 1.3mm to 1.3cm, with a size that is roughly proportional to wavelength. We discuss implications for models of Sgr A*, for theories of interstellar turbulence, and for imaging Sgr A* with the Event Horizon Telescope.Comment: 26 Pages, 17 Figures, Accepted for publication in Ap