Genetic Dissection of Endocytic Trafficking in Drosophila Using a Horseradish Peroxidase-Bride of Sevenless Chimera: hook Is Required for Normal Maturation of Multivesicular Endosomes

Mutations in the hook gene alter intracellular trafficking of internalized ligands in Drosophila. To dissect this defect in more detail, we developed a new approach to visualize the pathway taken by the Bride of Sevenless (Boss) ligand after its internalization into R7 cells. A chimeric protein consisting of HRP fused to Boss (HRP-Boss) was expressed in R8 cells. This chimera was fully functional: it rescued the boss mutant phenotype, and its trafficking was indistinguishable from that of the wild-type Boss protein. The HRP activity of the chimera was used to follow HRP-Boss trafficking on the ultrastructural level through early and late endosomes in R7 cells. In both wild-type and hook mutant eye disks, HRP-Boss was internalized into R7 cells. In wild-type tissue, Boss accumulated in mature multivesicular bodies (MVBs) within R7 cells; such accumulation was not observed in hook eye disks, however. Quantitative electron microscopy revealed a loss of mature MVBs in hook mutant tissue compared with wild type, whereas more than twice as many multilammelar late endosomes were detected. Our genetic analysis indicates that Hook is required late in endocytic trafficking to negatively regulate delivery from mature MVBs to multilammelar late endosomes and lysosomes.

Cloning and Characterization of peter pan, a Novel Drosophila Gene Required for Larval Growth

We identified a new Drosophila gene, peter pan (ppan), in a screen for larval growth–defective mutants. ppan mutant larvae do not grow and show minimal DNA replication but can survive until well after their heterozygotic siblings have pupariated. We cloned the ppan gene by P-element plasmid rescue. ppan belongs to a highly conserved gene family that includes Saccharomyces cerevisiae SSF1 and SSF2, as well as Schizosaccharomyces pombe, Arabidopsis, Caenorhabditis elegans, mouse, and human homologues. Deletion of both SSF1 and SSF2 in yeast is lethal, and depletion of the gene products causes cell division arrest. Mosaic analysis of ppan mutant clones in Drosophila imaginal disks and ovaries demonstrates that ppan is cell autonomous and required for normal mitotic growth but is not absolutely required for general biosynthesis or DNA replication. Overexpression of the wild-type gene causes cell death and disrupts the normal development of adult structures. The ppan gene family appears to have an essential and evolutionarily conserved role in cell growth.

The star formation history of CALIFA galaxies: Radial structures

We have studied the radial structure of the stellar mass surface density (μ∗) and stellar population age as a function of the total stellar mass and morphology for a sample of 107 galaxies from the CALIFA survey. We applied the fossil record method based on spectral synthesis techniques to recover the star formation history (SFH), resolved in space and time, in spheroidal and disk dominated galaxies with masses from 10^9 to 10^12 M_⊙. We derived the half-mass radius, and we found that galaxies are on average 15% more compact in mass than in light. The ratio of half-mass radius to half-light radius (HLR) shows a dual dependence with galaxy stellar mass; it decreases with increasing mass for disk galaxies, but is almost constant in spheroidal galaxies. In terms of integrated versus spatially resolved properties, we find that the galaxy-averaged stellar population age, stellar extinction, and μ_∗ are well represented by their values at 1 HLR. Negative radial gradients of the stellar population ages are present in most of the galaxies, supporting an inside-out formation. The larger inner (≤1 HLR) age gradients occur in the most massive (10^11 M_⊙) disk galaxies that have the most prominent bulges; shallower age gradients are obtained in spheroids of similar mass. Disk and spheroidal galaxies show negative μ∗ gradients that steepen with stellar mass. In spheroidal galaxies, μ∗ saturates at a critical value (~7 × 10^2 M_⊙/pc^2 at 1 HLR) that is independent of the galaxy mass. Thus, all the massive spheroidal galaxies have similar local μ_∗ at the same distance (in HLR units) from the nucleus. The SFH of the regions beyond 1 HLR are well correlated with their local μ_∗, and follow the same relation as the galaxy-averaged age and μ_∗; this suggests that local stellar mass surface density preserves the SFH of disks. The SFH of bulges are, however, more fundamentally related to the total stellar mass, since the radial structure of the stellar age changes with galaxy mass even though all the spheroid dominated galaxies have similar radial structure in μ_∗. Thus, galaxy mass is a more fundamental property in spheroidal systems, while the local stellar mass surface density is more important in disks.

Cold DUst around NEarby Stars (DUNES). First results. A resolved exo-Kuiper belt around the solar-like star ζ2 Ret

We present the first far-IR observations of the solar-type stars δ Pav, HR 8501, 51 Peg and ζ^2 Ret, taken within the context of the DUNES Herschel open time key programme (OTKP). This project uses the PACS and SPIRE instruments with the objective of studying infrared excesses due to exo-Kuiper belts around nearby solar-type stars. The observed 100 μm fluxes from δ Pav, HR 8501, and 51 Peg agree with the predicted photospheric fluxes, excluding debris disks brighter than L_dust/L_* ~ 5 x 10^-7 (1σ level) around those stars. A flattened, disk-like structure with a semi-major axis of ~100 AU in size is detected around ζ2 Ret. The resolved structure suggests the presence of an eccentric dust ring, which we interpret as an exo-Kuiper belt with L_dust/L_* ≈ 10^-5.

Caught in the act: gas and stellar velocity dispersions in a fast Quenching compact star-forming galaxy at z ~ 1.7

We present Keck I MOSFIRE spectroscopy in the Y and H bands of GDN-8231, a massive, compact, star-forming galaxy at a redshift of z ~ 1.7. Its spectrum reveals both Hα and [Nii] emission lines and strong Balmer absorption lines. The Hα and Spitzer MIPS 24 μm fluxes are both weak, thus indicating a low star-formation rate of SFR≲5-10 M_⨀ yr−1. This, added to a relatively young age of ~700 Myr measured from the absorption lines, provides the first direct evidence for a distant galaxy being caught in the act of rapidly shutting down its star formation. Such quenching allows GDN-8231 to become a compact, quiescent galaxy, similar to three other galaxies in our sample, by z ~ 1.5. Moreover, the color profile of GDN-8231 shows a bluer center, consistent with the predictions of recent simulations for an early phase of inside-out quenching. Its line-of-sight velocity dispersion for the gas, σ_LOG^gas = 127 ± 32 km s^−1, is nearly 40% smaller than that of its stars, σ_LOG^* = 215 ± 35 km s^−1. High-resolution hydro-simulations of galaxies explain such apparently colder gas kinematics of up to a factor of ~1.5 with rotating disks being viewed at different inclinations and/or centrally concentrated star-forming regions. A clear prediction is that their compact, quiescent descendants preserve some remnant rotation from their star-forming progenitors.

Avaliação de oleuropeína e de sanitizantes químicos, isolados ou associados, para eliminação de biofilmes de Staphylococcus aureus, Listeria monocytogenes e Escherichia coli em superfícies inertes

Este estudo avaliou a eficiência da oleuropeína (OLE) (composto fenólico extraído das folhas de Oliveira) isolada e associada aos sanitizantes comerciais ácido peracético 2% (APA), hipoclorito de sódio 2% (HS), peróxido de hidrogênio 3% (PH), digluconato de clorexidina 2% (DC), cloreto de benzalcônio 1% (CB) e iodofor 2% (IO), para inativação de células em suspensão e biofilmes monoespécie e multiespécie formados em superfícies de aço inoxidável ou microplaca de poliestireno por Listeria monocytogenes (ATCC 7644), Staphylococcus aureus (ATCC 25923) e Escherichia coli (ATCC 25922), todas classificadas como fortes produtores de biofilmes. Os isolados foram semeados em caldo TSB (caldo tripticase soja), incubados (37°C/24h) e corrigidos a ~108células/mL (escala 0,5 McFarland). Para bactérias em suspensão, a resistência a sanitizantes foi determinada pela Concentração Inibitória Mínima (CIM) em tubos e pelo método de Disco Difusão em Ágar (DDA), no qual as bactérias foram plaqueadas em ágar TSA contendo discos de 6mm de papel filtro embebidos nos sanitizantes. Após a incubação, a medição dos halos de inibição foi feita com paquímetro. Para os ensaios de resistência dos biofilmes aos compostos sanitizantes, foram utilizadas microplacas de poliestireno 96 poços, as quais foram preparadas para incubação-fixação dos biofilmes e submetidas à leitura em espectrofotômetro de ELISA (600 nm). Em seguida, as placas foram lavadas com solução salina tamponada (PBS, pH 7.4) e os sanitizantes inseridos por 1 minuto. Após neutralização com tiossulfato de sódio (5 minutos), as placas foram lavadas com PBS e metanol, coradas com cristal violeta 1% e coradas com ácido acético glacial (33%) para nova leitura a 570nm. A eficácia da remoção do biofilme pelos sanitizantes foi comparada pelo índice de formação de biofilme (IFB). As imagens do aço inoxidável após tratamento com sanitizante foram feitas através de Microscopia Eletrônica de Varredura (MEV) e Microscopia Confocal, para visualizar a persistência dos biofilmes. Os valores de CIM (diluição 1:2) mostraram que OLE não teve atividade bactericida. No método DDA, L. monocytogenes, foi resistente à OLE, enquanto E. coli e S. aureus apresentaram resistência intermediária. Os sanitizantes comerciais apresentaram boa atividade bactericida nos ensaios de CIM e DDA, sendo que as associações de OLE aos sanitizantes comerciais aumentaram o efeito germicida. Nos ensaios com biofilmes em monoespécie, somente os sanitizantes comerciais, isolados ou associados com OLE, foram eficazes de reduzir o valor de BFI em microplaca de poliestireno. Em biofilmes multiespécie, OLE apresentou efeito antimicrobiano, sobretudo sobre a associação de L. monocytogenes + E. coli + S. aureus (redução: 91,49%). Nenhum dos compostos avaliados foi capaz de inativar completamente os biofilmes nas superfícies de aço inoxidável, uma vez que células viáveis foram observadas após os tratamentos com os sanitizantes, indicando persistência dos biofilmes. Os resultados indicam que a oleuropeína apresentou potencial para incrementar o efeito bactericida de sanitizantes comerciais para eliminação de biofilmes em superfícies inertes, sendo necessários estudos para compreender os mecanismos de ação dessas combinações.

How I Learned to Stop Worrying and Love the Bits: Archival Motivation in the Digital Age

Why do people become archivists? Historically (and anecdotally) it was a deep love of musty, old records that drew people to the profession. While there have been many other motivating forces that inspired would-be archivists, it is most often that one hears of people seeking jobs in archives for love of “the stuff,” as evidenced in Kate Thiemer’s blog post, Honest tips for wannabe archivists (2012). As a result of the continually advancing presence of digitized and born digital archival collections, the physical nature of archival “stuff” is changing. While there remains the physical imprint of digital information on floppy disks, CDs, DVDs, hard drives, and old computers; the aspects of these physical artifacts might not evoke the same visceral pull to the profession as musty, raspy, paper-based documents. In light of this shift in physical presentation of information, we are faced with the question: how does love of archival “stuff” translate to work in digital archives? What is and/or will be the pull to become a digital archivist? To answer these questions, we will perform a survey-based study where we will invite archivists who work with both traditional and digital archival material to answer questions related to the aspects of their work that inspired or motivated them to join the profession. What motivates people to become archivists? What aspects of digital archives do or can potentially motivate people to seek out a career as an archivist? What, if any, motivational factors for becoming a traditional archivist are the same as those for becoming a digital archivist? What, if any, motivational factors for becoming a traditional archivist are different from those for becoming a digital archivist? By answering these questions, we hope to expand the archival discussion on what it means to be an archivist in the digital age. What compelling intrinsic, evidential, or informational values are present in digital archival content that will draw professionals to the field? Are there other values inherent in digital content that are currently unexplored? In our poster, we will present our discussion of the topic, our survey design, and results we have at the time of the Institute. Thiemer, K. (2012). Honest tips for wannabe archivists. Archivesnext blog. Retrieved from http://www.archivesnext.com/?p=2849

GRO J1008−57: an (almost) predictable transient X-ray binary

A study of archival RXTE, Swift, and Suzaku pointed observations of the transient high-mass X-ray binary GRO J1008−57 is presented. A new orbital ephemeris based on pulse arrival-timing shows the times of maximum luminosities during outbursts of GRO J1008−57 to be close to periastron at orbital phase − 0.03. This makes the source one of a few for which outburst dates can be predicted with very high precision. Spectra of the source in 2005, 2007, and 2011 can be well described by a simple power law with high-energy cutoff and an additional black body at lower energies. The photon index of the power law and the black-body flux only depend on the 15–50 keV source flux. No apparent hysteresis effects are seen. These correlations allow us to predict the evolution of the pulsar’s X-ray spectral shape over all outbursts as a function of just one parameter, the source’s flux. If modified by an additional soft component, this prediction even holds during GRO J1008−57’s 2012 type II outburst.

Trajectory-Based Morphological Operators: A Model for Efficient Image Processing

Mathematical morphology has been an area of intensive research over the last few years. Although many remarkable advances have been achieved throughout these years, there is still a great interest in accelerating morphological operations in order for them to be implemented in real-time systems. In this work, we present a new model for computing mathematical morphology operations, the so-called morphological trajectory model (MTM), in which a morphological filter will be divided into a sequence of basic operations. Then, a trajectory-based morphological operation (such as dilation, and erosion) is defined as the set of points resulting from the ordered application of the instant basic operations. The MTM approach allows working with different structuring elements, such as disks, and from the experiments, it can be extracted that our method is independent of the structuring element size and can be easily applied to industrial systems and high-resolution images.

Deriving the Dynamical Masses of Ultra-Luminous X-ray Sources with OSIRIS

Ultra Luminous X-ray Sources (ULXs) are extragalactic X-ray point sources with LX ∼ 1039 − 1041 erg s−1 discovered in the 80s with the Einstein satellite and confirmed as black hole X-ray binaries during the last decade. The nature of the compact object is highly controversial. They could be super-Eddington stellar-mass black holes or intermediate mass black holes. Deriving dynamical masses of the brightest ULXs, which can be done with OSIRIS, is the only way to find out the nature of the compact object.

Screening Subclinical Keratoconus With Placido-Based Corneal Indices

Purpose. To assess in a sample of normal, keratoconic, and keratoconus (KC) suspect eyes the performance of a set of new topographic indices computed directly from the digitized images of the Placido rings. Methods. This comparative study was composed of a total of 124 eyes of 106 patients from the ophthalmic clinics Vissum Alicante and Vissum Almería (Spain) divided into three groups: control group (50 eyes), KC group (50 eyes), and KC suspect group (24 eyes). In all cases, a comprehensive examination was performed, including the corneal topography with a Placidobased CSO topography system. Clinical outcomes were compared among groups, along with the discriminating performance of the proposed irregularity indices. Results. Significant differences at level 0.05 were found on the values of the indices among groups by means of Mann-Whitney-Wilcoxon nonparametric test and Fisher exact test. Additional statistical methods, such as receiver operating characteristic analysis and K-fold cross validation, confirmed the capability of the indices to discriminate between the three groups. Conclusions. Direct analysis of the digitized images of the Placido mires projected on the cornea is a valid and effective tool for detection of corneal irregularities. Although based only on the data from the anterior surface of the cornea, the new indices performed well even when applied to the KC suspect eyes. They have the advantage of simplicity of calculation combined with high sensitivity in corneal irregularity detection and thus can be used as supplementary criteria for diagnosing and grading KC that can be added to the current keratometric classifications.

Measuring the stellar wind parameters in IGR J17544-2619 and Vela X-1 constrains the accretion physics in supergiant fast X-ray transient and classical supergiant X-ray binaries

Context. Classical supergiant X-ray binaries (SGXBs) and supergiant fast X-ray transients (SFXTs) are two types of high-mass X-ray binaries (HMXBs) that present similar donors but, at the same time, show very different behavior in the X-rays. The reason for this dichotomy of wind-fed HMXBs is still a matter of debate. Among the several explanations that have been proposed, some of them invoke specific stellar wind properties of the donor stars. Only dedicated empiric analysis of the donors’ stellar wind can provide the required information to accomplish an adequate test of these theories. However, such analyses are scarce. Aims. To close this gap, we perform a comparative analysis of the optical companion in two important systems: IGR J17544-2619 (SFXT) and Vela X-1 (SGXB). We analyze the spectra of each star in detail and derive their stellar and wind properties. As a next step, we compare the wind parameters, giving us an excellent chance of recognizing key differences between donor winds in SFXTs and SGXBs. Methods. We use archival infrared, optical and ultraviolet observations, and analyze them with the non-local thermodynamic equilibrium (NLTE) Potsdam Wolf-Rayet model atmosphere code. We derive the physical properties of the stars and their stellar winds, accounting for the influence of X-rays on the stellar winds. Results. We find that the stellar parameters derived from the analysis generally agree well with the spectral types of the two donors: O9I (IGR J17544-2619) and B0.5Iae (Vela X-1). The distance to the sources have been revised and also agree well with the estimations already available in the literature. In IGR J17544-2619 we are able to narrow the uncertainty to d = 3.0 ± 0.2 kpc. From the stellar radius of the donor and its X-ray behavior, the eccentricity of IGR J17544-2619 is constrained to e< 0.25. The derived chemical abundances point to certain mixing during the lifetime of the donors. An important difference between the stellar winds of the two stars is their terminal velocities (ν∞ = 1500 km s-1 in IGR J17544-2619 and ν∞ = 700 km s-1 in Vela X-1), which have important consequences on the X-ray luminosity of these sources. Conclusions. The donors of IGR J17544-2619 and Vela X-1 have similar spectral types as well as similar parameters that physically characterize them and their spectra. In addition, the orbital parameters of the systems are similar too, with a nearly circular orbit and short orbital period. However, they show moderate differences in their stellar wind velocity and the spin period of their neutron star which has a strong impact on the X-ray luminosity of the sources. This specific combination of wind speed and pulsar spin favors an accretion regime with a persistently high luminosity in Vela X-1, while it favors an inhibiting accretion mechanism in IGR J17544-2619. Our study demonstrates that the relative wind velocity is critical in class determination for the HMXBs hosting a supergiant donor, given that it may shift the accretion mechanism from direct accretion to propeller regimes when combined with other parameters.

Using new scanning electron microscopy (SEM) approaches to elucidate bacterial biofilm architecture

Healthcare-associated infections (HAI) are a major public health problem being Klebsiella pneumoniae and nontuberculous mycobacteria, both with high antibiotic resistance rates, among their etiological agent. Since biofilme assembly is pointed as one of the mechanisms involved in emergence of antibiotic resistance understanding bacteria organization within the biofilm and the identification of differences between planktonic and sessile forms of bacteria will be a step forward to fight HAI. In the present work we used SEM as a tool to characterize the internal structure of biofilm assembled on different surfaces. For SEM analysis, biofilms were allowed to form either on six-well cell culture plates, silicon or metallic disks placed inside the wells for different incubation periods at 37 °C. The biofilm assembled on the cell culture dish was for both secondary and backscattered electron analysis as described before. Biofilms assembled on silicon disks instead of being sectioned were prepared as metallographic samples, by grinding with grit SIC paper and polishing with diamond particles. Samples were cleaned (70% ethanol), dried with hot air, further coated and analysed. A preliminary study using FIB-SEM has been performed to access the ultrastructure of biofilms assembled on metallic surfaces. The results obtained showed that the same bacteria assembled biofilms with different ratios of biomass and extracellular matrix depending on the surface. SEM performed on thin sections of biofilms is a powerful tool to elucidate biofilm structure allowing the quantification of the major components. FIB-SEM is also a promising tool in this field.

ICR water utility database system users' guide /

"EPA 814/B-96-004."

“How Do You ‘Do’ Bokeh?” The Aesthetics of How-to Videos about Background Blur and Eroticized Women

Producers of online instructional videos about bokeh emphasize disks of light and out-of-focus backgrounds. They demonstrate how camera lenses and technical features can render bokeh, or unfocused areas. Photographic and video bokeh ordinarily appears away from the center of attention. The bokeh genre, in opposition to typical photography and video practices, foregrounds the peripheral and proposes aesthetics and ways of looking by seeing and not seeing objects. However, producers of online instructional videos about bokeh sometimes couple their sensual aestheticization of backgrounds to their stated attempts to satisfy viewers’ investments in filling foregrounds with images of objectified women. These producers emphasize unconventional aesthetics as a means of establishing their creative and technical expertise and obscuring their reproduction of traditional conceptions of women as viewable and controllable. Close textual analysis, literature on photography and transparency, and feminist considerations of representation allow me to consider the aesthetics and functions of this how-to form.