Factors associated with condom use among HIV-positive women living in Atlanta, Georgia.

OBJECTIVES:Consistent condom use is essential to reducing heterosexual transmission of HIV. African Americans are disproportionately affected by HIV in the United States despite comprising a small percentage of the population. Our objectives were to evaluate factors associated with self-reported condom use in a cohort of predominantly African American women receiving HIV care in Atlanta, Georgia. METHODS:A cross-sectional study of reproductive knowledge, attitudes, and practices among adult, sexually-active, HIV-positive women attending the Grady Infectious Disease Clinic in Atlanta, Georgia was conducted from July, 2013 to November, 2014 to evaluate factors associated with self-reported condom use. Primary outcomes included: condom use at last vaginal intercourse and consistent condom use with vaginal intercourse over the last six months. Descriptive, bivariable, and multivariable logistic regression analyses were performed. RESULTS:Of 187 women enrolled, 170 reported having vaginal intercourse in the last six months. Seventy-four percent used condoms at last vaginal intercourse, whereas 53% reported consistent condom use over the last six months. In adjusted analyses, factors associated with condom use at last intercourse included decreased frequency of sex, no history of drug use, and confidence to discuss condom use with sexual partners (p<0.05). Factors associated with consistent condom use in the past six months were older age, being single/dating, and confidence to discuss condom use with sexual partners. History of drug use, having HIV-positive partners, and unprotected anal intercourse were associated with inconsistent use (p<0.05). CONCLUSIONS:Improved strategies are needed to educate women on the importance of safe sexual practices and condom negotiation. Healthcare providers should strive to have an open dialogue with patients about condom use, whether they engage in anal sex, and its risks

Novel role of the muskelin-RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation

The evolutionarily conserved kelch-repeat protein muskelin was identified as an intracellular mediator of cell spreading. We discovered that its morphological activity is controlled by association with RanBP9/RanBPM, a protein involved in transmembrane signaling and a conserved intracellular protein complex. By subcellular fractionation, endogenous muskelin is present in both the nucleus and the cytosol. Muskelin subcellular localization is coregulated by its C terminus, which provides a cytoplasmic restraint and also controls the interaction of muskelin with RanBP9, and its atypical lissencephaly-1 homology motif, which has a nuclear localization activity which is regulated by the status of the C terminus. Transient or stable short interfering RNA–based knockdown of muskelin resulted in protrusive cell morphologies with enlarged cell perimeters. Morphology was specifically restored by complementary DNAs encoding forms of muskelin with full activity of the C terminus for cytoplasmic localization and RanBP9 binding. Knockdown of RanBP9 resulted in equivalent morphological alterations. These novel findings identify a role for muskelin–RanBP9 complex in pathways that integrate cell morphology regulation and nucleocytoplasmic communication

(A) Localization of GFP-MK and the panel of muskelin domain deletions, as shown in , visualized in fixed COS-7 cells by laser scanning confocal microscopy

Merged images show GFP proteins (green) and DAPI-stained nuclei (blue). For HA-DDL, anti-HA staining is green. Images are in the xy plane unless otherwise indicated. Exclusion of nuclear-located MKΔC35 from nucleoli is demonstrated in the inset and xz panels as merged images of GFP-MKΔC35 (green) and anti-fibrillarin (red) staining. Bars, 10 μm. (B) Quantification of the subcellular localizations of GFP-tagged muskelin domain deletion proteins by a five-point scoring scheme. The major localization of each protein is also shown schematically. All localizations were scored blind by two independent observers. Each column represents the mean from three to five independent experiments. Error bars indicate SEM. At least 1,000 cells were scored for each construct.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p

(A–D) siRNA-based depletion of muskelin leads to morphological alterations

(A) Reduced attachment of A549 cells with stable expression of muskelin-shRNA clone 2 (cl2) and clone 3 (cl3) cells to the TSP-1 C-terminal protein T3GCT. Adhesion assays were performed for 1 h under serum-free conditions as previously described (). (B and C) Cell perimeter measurements for control and muskelin-depleted A549 (B) or C2C12 (C) cells adherent on 50 nM FN for 1 h. (B) A549 lines were also compared after 48 h expression of GFP or GFP-MK. Only GFP-MK restored normal morphology. In each graph, each column represents the mean of three independent experiments. Error bars indicate SEM. *, P ≤ 0.0001. (D) Representative F-actin organization in control and muskelin-depleted C2C12 cells under the same experimental conditions as in C. Bar, 10 μm. (E) Activity of muskelin mutants in restoring the morphology of muskelin-depleted A549 clone 3 cells. GFP, GFP-MK, or the indicated mutants were expressed in clone 3 cells for 48 h, the cells adhered to 50 nM FN for 1 h, and cell perimeter lengths of GFP-positive cells were measured. Each column represents the mean of three independent experiments. Error bars indicate SEM. *, P ≤ 0.00001. At least 500 cells were scored per condition. (F) RanBP9 knockdown results in similar morphological changes in FN-adherent A549 cells. Where indicated, cells were treated with 0.75 μg/ml doxycycline for 36 h. Each column represents the mean from >50 cells. Error bars indicate SEM. *, P < 0.0001. Insets show representative merged cell images of F-actin (green), DAPI (blue), and, where induced, shRNA transcript expression (red; RFP). Bar, 10 μm.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p

(A) Schematic diagram of the FLAG-tagged RanBP9 domains prepared for these experiments

SPRY, Sp1a and ryanodine receptor domain; L, LisH motif; H, CTLH motif; u, unique region; CRA, CT11/RanBP9 domain. (B) Specific coimmunoprecipitation of FLAG-RanBP9[142–748] with GFP-MK. (C) Analysis of RanBP9 domain requirements for coimmunoprecipitation with GFP-MK. The boxed inset is a longer exposure of three lysate lanes demonstrating the poorly expressed LHuCRA and HuCRA proteins. Asterisks indicate observed weak specific coimmunoprecipitations. All molecular mass markers are shown in kilodaltons. Results shown are representative of three experiments.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p

(A) Localization of GFP-MK in COS-7 cells treated with solvent (Con) or 10 μg/ml leptomycin B for 18 h

Bar, 10 μm. (B) Quantification of the cellular localizations of GFP-MK or cyclin B1 in cells treated with solvent or 10 μg/ml leptomycin B for 18 h. Only the distribution of cyclin B1 was significantly altered (*, P < 0.001). Each column represents the mean from three independent experiments. Error bars indicate SEM. At least 100 cells were scored in each experiment.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p

(A) Schematic diagram of RanBP9 domain architecture and the domains included in the yeast two-hybrid clone

(B) Localization of RanBP9 in subcellular fractions of C2C12 cells. Cy, cytosol; M, membranes; NE, nuclear extract; NP, nuclear pellet. (C) The domains sufficient for coimmunoprecipitation of endogenous RanBP9 with GFP-MK are included in the LHKC fragment. (D) Analysis of the minimal domain requirements within the LHKC domains for coimmunoprecipitation of endogenous RanBP9. (E and F) Coimmunoprecipitation of endogenous RanBP9 by GFP-MK is regulated by the status of muskelin C35 region. (F) RanBP9 signal intensities normalized relative to GFP-MK immunoprecipitation are given below the blot. All molecular mass markers are shown in kilodaltons. Results shown are representative of three experiments.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p

(A and B) Immunoblots demonstrate equivalent expression of GFP-MK and the indicated point mutant proteins

Molecular mass markers are shown in kilodaltons. (C–F) Merged confocal images showing localization of GFP-tagged muskelin point mutants (green) in COS-7 cells costained with DAPI (blue). Bars, 10 μm. (G) Quantification of subcellular localizations. Each column represents the mean from two to three independent experiments. Error bars indicate SEM. At least 500 cells were scored for each construct. The localization of each protein is also shown schematically.<p><b>Copyright information:</b></p><p>Taken from "Novel role of the muskelin–RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation"</p><p></p><p>The Journal of Cell Biology 2008;182(4):727-739.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518711.</p><p></p