Interactions in vivo between the Vif protein of HIV-1 and the precursor (Pr55GAG) of the virion nucleocapsid proteins

The abnormality of viral core structure seen in vif-defective HIV-1 grown in PBMCs has suggested a role for Vif in viral morphogenesis. Using an in vivo mammalian two-hybrid assay, the interaction between Vif and the precursor (Pr55GAG) of the virion nucleocapsid proteins has been analysed. This revealed the amino-terminal (aa 1–22) and central (aa 70–100) regions of Vif to be essential for its interaction with Pr55GAG, but deletion of the carboxy-terminal (aa 158–192) region of the protein had only a minor effect on its interaction. Initial deletion studies carried out on Pr55GAG showed that a 35-amino-acid region of the protein bridging the MA(p17)–CA(p24) junction was essential for its ability to interact with Vif. Site-directed mutagenesis of a conserved tryptophan (Trp21) near the amino terminus of Vif showed it to be important for the interaction with Pr55GAG. By contrast, mutagenesis of the highly conserved YLAL residues forming part of the BC-box motif, shown to be important in Vif promoting degradation of APOBEC3G/3F, had little or no effect on the Vif–Pr55GAG interaction

Hubble Space Telescope NICMOS Polarization Observations of Three Edge-on Massive YSOs

Massive young stellar objects (YSOs), like low-mass YSOs, appear to be surrounded by optically thick envelopes and/or disks and have regions, often bipolar, that are seen in polarized scattered light at near-infrared wavelengths. We are using the 0.2'' spatial resolution of NICMOS on Hubble Space Telescope to examine the structure of the disks and outflow regions of massive YSOs in star-forming regions within a few kpc of the Sun. Here we report on 2 micron polarimetry of NGC 6334 V and S255 IRS1. NGC 6334 V consists of a double-lobed bright reflection nebula seen against a dark region, probably an optically thick molecular cloud. Our polarization measurements show that the illuminating star lies ~ 2'' south of the line connecting the two lobes; we do not detect this star at 2 micron, but there are a small radio source and a mid-infrared source at this location. S255 IRS1 consists of two YSOs (NIRS1 and NIRS3) with overlapping scattered light lobes and luminosities corresponding to early B stars. Included in IRS1 is a cluster of stars from whose polarization we determine the local magnetic field direction. Neither YSO has its scattered light lobes aligned with this magnetic field. The line connecting the scattered light lobes of NIRS1 is twisted symmetrically around the star; the best explanation is that the star is part of a close binary and the outflow axis of NIRS1 is precessing as a result of non-coplanar disk and orbit. The star NIRS3 is also offset from the line connecting its two scattered light lobes. We suggest that all three YSOs show evidence of episodic ejection of material as they accrete from dense, optically thick envelopes.Comment: 39 pages, 7 figures, 4 tables To be published in The Astrophysical Journa

Сутність та класифікація ризиків інвестиційної діяльності

Наводиться визначення поняттю "ризики інвестиційної діяльності" за рахунок поєднання його сутнісних характеристик, виконано узагальнення класифікації цих ризиків, запропоновано введення нової класифікаційної групи – "корпоративні ризики", які пов'язані з можливістю втрати контролю над підприємством інвестором-акціонером

Multiomic profiling of breast cancer cells uncovers stress MAPK-associated sensitivity to AKT degradation

More than 50% of human tumors display hyperactivation of the serine/threonine kinase AKT. Despite evidence of clinical efficacy, the therapeutic window of the current generation of AKT inhibitors could be improved. Here, we report the development of a second-generation AKT degrader, INY-05-040, which outperformed catalytic AKT inhibition with respect to cellular suppression of AKT-dependent phenotypes in breast cancer cell lines. A growth inhibition screen with 288 cancer cell lines confirmed that INY-05-040 had a substantially higher potency than our first-generation AKT degrader (INY-03-041), with both compounds outperforming catalytic AKT inhibition by GDC-0068. Using multiomic profiling and causal network integration in breast cancer cells, we demonstrated that the enhanced efficacy of INY-05-040 was associated with sustained suppression of AKT signaling, which was followed by induction of the stress mitogen-activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK). Further integration of growth inhibition assays with publicly available transcriptomic, proteomic, and reverse phase protein array (RPPA) measurements established low basal JNK signaling as a biomarker for breast cancer sensitivity to AKT degradation. Together, our study presents a framework for mapping the network-wide signaling effects of therapeutically relevant compounds and identifies INY-05-040 as a potent pharmacological suppressor of AKT signaling

Multiomic profiling of breast cancer cells uncovers stress MAPK-associated sensitivity to AKT degradation

More than 50% of human tumors display hyperactivation of the serine/threonine kinase AKT. Despite evidence of clinical efficacy, the therapeutic window of the current generation of AKT inhibitors could be improved. Here, we report the development of a second-generation AKT degrader, INY-05-040, which outperformed catalytic AKT inhibition with respect to cellular suppression of AKT-dependent phenotypes in breast cancer cell lines. A growth inhibition screen with 288 cancer cell lines confirmed that INY-05-040 had a substantially higher potency than our first-generation AKT degrader (INY-03-041), with both compounds outperforming catalytic AKT inhibition by GDC-0068. Using multiomic profiling and causal network integration in breast cancer cells, we demonstrated that the enhanced efficacy of INY-05-040 was associated with sustained suppression of AKT signaling, which was followed by induction of the stress mitogen-activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK). Further integration of growth inhibition assays with publicly available transcriptomic, proteomic, and reverse phase protein array (RPPA) measurements established low basal JNK signaling as a biomarker for breast cancer sensitivity to AKT degradation. Together, our study presents a framework for mapping the network-wide signaling effects of therapeutically relevant compounds and identifies INY-05-040 as a potent pharmacological suppressor of AKT signaling

Strong CH+ J=1-0 emission and absorption in DR21

We report the first detection of the ground-state rotational transition of the methylidyne cation CH+ towards the massive star-forming region DR21 with the HIFI instrument onboard the Herschel satellite. The line profile exhibits a broad emission line, in addition to two deep and broad absorption features associated with the DR21 molecular ridge and foreground gas. These observations allow us to determine a CH+ J=1-0 line frequency of 835137 +/- 3 MHz, in good agreement with a recent experimental determination. We estimate the CH+ column density to be a few 1e13 cm^-2 in the gas seen in emission, and > 1e14 cm^-2 in the components responsible for the absorption, which is indicative of a high line of sight average abundance [CH+]/[H] > 1.2x10^-8. We show that the CH+ column densities agree well with the predictions of state-of-the-art C-shock models in dense UV-illuminated gas for the emission line, and with those of turbulent dissipation models in diffuse gas for the absorption lines.Comment: Accepted for publication in A&

Dynamics in the satellite system of Triangulum: Is AndXXII a dwarf satellite of M33?

We present results from a spectroscopic survey of the dwarf spheroidal And XXII and the two extended clusters EC1 and EC2. These three objects are candidate satellites of the Triangulum galaxy, M33, which itself is likely a satellite of M31. We use the DEep Imaging Multi-Object Spectrograph mounted on the Keck-II telescope to derive radial velocities for candidate member stars of these objects and thereby identify the stars that are most likely actual members. Eleven most probable stellar members (of 13 candidates) are found for AndXXII. We obtain an upper limit of sigma_v < 6.0 km s-1 for the velocity dispersion of AndXXII, [Fe/H] ~ -1.6 for its metallicity, and 255pc for the Plummer radius of its projected density profile. We construct a colour magnitude diagram for AndXXII and identify both the red giant branch and the horizontal branch. The position of the latter is used to derive a heliocentric distance to And XXII of 853 pm 26 kpc. The combination of the radial velocity, distance, and angular position of AndXXII indicates that it is a strong candidate for being the first known satellite of M33 and one of the very few examples of a galactic satellite of a satellite. N-body simulations imply that this conclusion is unchanged even if M31 and M33 had a strong encounter in the past few Gyr. We test the hypothesis that the extended clusters highlight tidally stripped galaxies by searching for an excess cloud of halo-like stars in their vicinity. We find such a cloud for the case of EC1 but not EC2. The three objects imply a dynamical mass for M33 that is consistent with previous estimates.Comment: 14 pages, 14 figures, revised for MNRAS publicatio

The Vital Role of Social Workers in Community Partnerships: The Alliance for Gay, Lesbian, Bisexual, Transgender and Questioning Youth

The account of The Alliance for Gay, Lesbian, Bisexual, Transgender, and Questioning (GLBTQ) Youth formation offers a model for developing com- munity-based partnerships. Based in a major urban area, this university-community collaboration was spearheaded by social workers who were responsible for its original conceptualization, for generating community support, and for eventual staffing, administration, direct service provision, and program evaluation design. This article presents the strategic development and evolution of this community- based service partnership, highlighting the roles of schools of social work, academics, and social work students in concert with community funders, practitioners and youth, in responding to the needs of a vulnerable population

Green Fluorescent Protein Labeling of Listeria, Salmonella, and Escherichia coli O157:H7 for Safety-Related Studies

Many food safety-related studies require tracking of introduced foodborne pathogens to monitor their fate in complex environments. The green fluorescent protein (GFP) gene (gfp) provides an easily detectable phenotype so has been used to label many microorganisms for ecological studies. The objectives of this study were to label major foodborne pathogens and related bacteria, including Listeria monocytogenes, Listeria innocua, Salmonella, and Escherichia coli O157:H7 strains, with GFP and characterize the labeled strains for stability of the GFP plasmid and the plasmid's effect on bacterial growth. GFP plasmids were introduced into these strains by a CaCl2 procedure, conjugation or electroporation. Stability of the label was determined through sequential propagation of labeled strains in the absence of selective pressure, and rates of plasmid-loss were calculated. Stability of the GFP plasmid varied among the labeled species and strains, with the most stable GFP label observed in E. coli O157:H7. When grown in nonselective media for two consecutive subcultures (ca. 20 generations), the rates of plasmid loss among labeled E. coli O157:H7, Salmonella and Listeria strains ranged from 0%–30%, 15.8%–99.9% and 8.1%–93.4%, respectively. Complete loss (>99.99%) of the plasmid occurred in some labeled strains after five consecutive subcultures in the absence of selective pressure, whereas it remained stable in others. The GFP plasmid had an insignificant effect on growth of most labeled strains. E. coli O157:H7, Salmonella and Listeria strains can be effectively labeled with the GFP plasmid which can be stable in some isolates for many generations without adversely affecting growth rates

A random mutation capture assay to detect genomic point mutations in mouse tissue

Herein, a detailed protocol for a random mutation capture (RMC) assay to measure nuclear point mutation frequency in mouse tissue is described. This protocol is a simplified version of the original method developed for human tissue that is easier to perform, yet retains a high sensitivity of detection. In contrast to assays relying on phenotypic selection of reporter genes in transgenic mice, the RMC assay allows direct detection of mutations in endogenous genes in any mouse strain. Measuring mutation frequency within an intron of a transcribed gene, we show this assay to be highly reproducible. We analyzed mutation frequencies from the liver tissue of animals with a mutation within the intrinsic exonuclease domains of the two major DNA polymerases, δ and ε. These mice exhibited significantly higher mutation frequencies than did wild-type animals. A comparison with a previous analysis of these genotypes in Big Blue mice revealed the RMC assay to be more sensitive than the Big Blue assay for this application. As RMC does not require analysis of a particular gene, simultaneous analysis of mutation frequency at multiple genetic loci is feasible. This assay provides a versatile alternative to transgenic mouse models for the study of mutagenesis in vivo