The DUSP domain of pseudophosphatase MK-STYX interacts with G3BP1 to decrease stress granules.

Mitogen activated protein kinase phosphoserine/threonine/tyrosine-binding protein (MK-STYX) is a dual specificity (DUSP) member of the protein tyrosine phosphatase family. It is a pseudophosphatase, which lacks the essential amino acids histidine and cysteine in the catalytic active signature motif (HCX5R). We previously reported that MK-STYX interacts with G3BP1 [Ras-GAP (GTPase-activating protein) SH3 (Src homology 3) domain-binding-1] and reduces stress granules, stalled mRNA. To determine how MK-STYX reduces stress granules, truncated domains, CH2 (cell division cycle 25 phosphatase homology 2) and DUSP, of MK-STYX were used. Wild-type MK-STYX and the DUSP domain significantly decreased stressed granules that were induced by sodium arsenite, in which G3BP1 (a stress granule nucleator) was used as the marker. In addition, HEK/293 and HeLa cells co-expressing G3BP1-GFP and mCherry-MK-STYX, mCherry-MK-STYX-CH2, mCherry-MK-STYX-DUSP or mCherry showed that stress granules were significantly decreased in the presence of wild-type MK-STYX and the DUSP domain of MK-STYX. Further characterization of these dynamics in HeLa cells showed that the CH2 domain increased the number of stress granules within a cell, relative to wild-type and DUSP domain of MK-STYX. To further analyze the interaction of G3BP1 and the domains of MK-STYX, coimmunoprecipitation experiments were performed. Cells co-expressing G3BP1-GFP and mCherry, mCherry-MK-STYX, mCherry-MK-STYX-CH2, or mCherry-MK-STYX-DUSP demonstrated that the DUSP domain of MK-STYX interacts with both G3BP1-GFP and endogenous G3BP1, whereas the CH2 domain of MK-STYX did not coimmunoprecipitate with G3BP1. In addition, G3BP1 tyrosine phosphorylation, which is required for stress granule formation, was decreased in the presence of wild-type MK-STYX or the DUSP domain but increased in the presence of CH2. These data highlight a model for how MK-STYX decreases G3BP1-induced stress granules. The DUSP domain of MK-STYX interacts with G3BP1 and negatively alters its tyrosine phosphorylation- decreasing stress granule formation.

Nevirapine and zidovudine at birth to reduce perinatal transmission of HIV in an African setting: a randomized controlled trial.

CONTEXT: Antenatal counseling and human immunodeficiency virus (HIV) testing are not universal in Africa; thus, women often present in labor with unknown HIV status without receiving the HIVNET 012 nevirapine (NVP) regimen (a single oral dose of NVP to the mother at the start of labor and to the infant within 72 hours of birth). OBJECTIVE: To determine risk of mother-to-child transmission of HIV when either standard use of NVP alone or in combination with zidovudine (ZDV) was administered to infants of women tested at delivery. DESIGN, SETTING, AND PARTICIPANTS: A randomized, open-label, phase 3 trial conducted between April 1, 2000, and March 15, 2003, at 6 clinics in Blantyre, Malawi, Africa. The trial included all infants born to 894 women who were HIV positive, received NVP intrapartum, and were previously antiretroviral treatment-naive. Infants were randomly assigned to NVP (n = 448) and NVP plus ZDV (n = 446). Infants were enrolled at birth, observed at 6 to 8 weeks, and followed up through 3 to 18 months. The HIV status of 90% of all infants was established at 6 to 8 weeks. INTERVENTION: Mothers received a 200-mg single oral dose of NVP intrapartum and infants received either 2-mg/kg oral dose of NVP or NVP (same dose) plus 4 mg/kg of ZDV twice per day for a week. MAIN OUTCOME MEASURES: HIV infection of infant at birth and 6 to 8 weeks, and adverse events. RESULTS: The mother-to-child transmission of HIV at birth was 8.1% (36/445) in infants administered NVP only and 10.1% (45/444) in those administered NVP plus ZDV (P =.30). A life table estimate of transmission at 6 to 8 weeks was 14.1% (95% confidence interval [CI], 10.7%-17.4%) in infants who received NVP and 16.3% (95% CI, 12.7%-19.8%) in those who received NVP plus ZDV (P =.36). For infants not infected at birth and retested at 6 to 8 weeks, transmission was 6.5% (23/353) in those who received NVP only and 6.9% (25/363) in those who received NVP plus ZDV (P =.88). Almost all infants (99%-100%) were breastfed at 1 week and 6 to 8 weeks. Grades 3 and 4 adverse events were comparable; 4.9% (22/448) and 5.4% (24/446) in infants receiving NVP only and NVP plus ZDV, respectively (P =.76). CONCLUSIONS: The frequency of mother-to-child HIV transmission at 6 to 8 weeks in our 2 study groups was comparable with that observed for other perinatal HIV intervention studies among breastfeeding women in Africa. The safety of the regimen containing neonatal ZDV was similar to that of a standard NVP regimen.

Use of tablet-based kiosks in the emergency department to guide patient HIV self-testing with a point-of-care oral fluid test.

Despite successes in efforts to integrate HIV testing into routine care in emergency departments, challenges remain. Kiosk-facilitated, directed HIV self-testing offers one novel approach to address logistical challenges. Emergency department patients, 18-64 years, were recruited to evaluate use of tablet-based-kiosks to guide patients to conduct their own point-of-care HIV tests followed by standard-of-care HIV tests by healthcare workers. Both tests were OraQuick Advance tests. Of 955 patients approached, 473 (49.5%) consented; 467 completed the test, and 100% had concordant results with healthcare workers. Median age was 41 years, 59.6% were female, 74.8% were African-American, and 19.6% were White. In all, 99.8% of patients believed the self-test was "definitely" or "probably" correct; 91.7% of patients "trusted their results very much"; 99.8% reported "overall" self-testing was "easy or somewhat easy" to perform. Further, 96.9% indicated they would "probably" or "definitely" test themselves at home were the HIV test available for purchase; 25.9% preferred self-testing versus 34.4% who preferred healthcare professional testing (p>0.05). Tablet-based kiosk testing proved to be highly feasible, acceptable, and an accurate method of conducting rapid HIV self-testing in this study; however, rates of engagement were moderate. More research will be required to ascertain barriers to increased engagement for self-testing.

Patients Can Accurately Perform Their Own Rapid HIV Point-of-Care Test in the Emergency Department.

OBJECTIVE: The objective of this study was to evaluate the feasibility, acceptability, and accuracy of having emergency department (ED) patients perform a rapid, point-of-care (POC) self-test for HIV before routine HIV testing. METHODS: Patients aged 18 to 65 years were recruited to perform a rapid POC HIV oral fluid at The Johns Hopkins ED in conjunction with the standard-of-care HIV POC test. Acceptability and ease of use were assessed by a questionnaire. RESULTS: A total of 259 patients were approached for testing, and 249 (96.1%) consented to perform a self POC HIV test. Of patients performing a self-test, 100% had concordant results with those obtained by the health care worker. Four females (1.6%) were newly identified as HIV positive. Median participant age was 41 years, and 58% of patients were female; 83% were African American, and 16% were white. Overall, greater than 90% of patients reported trust of the test results, ease of testing, and willingness to test again. Approximately 35% of patients indicated they would pay up to a maximum price of $30 for testing. Overall, 46.9% of patients preferred self-testing, and 39.5% preferred health care professional testing. Regarding preferred location for testing, 51.0% preferred home self-testing, 39.5% preferred clinic/ED self-testing (P > 0.05), and 9.5% had no preference. CONCLUSIONS: A significant proportion of patients offered POC testing in the ED agreed to perform a self-HIV test. Patients' results were concordant with those obtained by the health care worker; 1.6% were HIV positive. The majority of participants believed the veracity of their results. A greater number of patients preferred self-testing.

The effect of human immunodeficiency virus and breastfeeding on the nutritional status of African children.

BACKGROUND: The risk of HIV-1 infection is high among breast-fed children in sub-Saharan Africa. Monitoring the nutritional status can provide useful information to determine the effect of HIV infection and breast-feeding on child growth and development. We longitudinally assessed the nutritional status and determined its association with HIV infection and breast-feeding among Malawian children. METHODS: We analyzed data from 2 clinical trials to prevent mother-to-child transmission of HIV in Malawi. These trials were conducted during 2000-2003 before the current guidelines were implemented to breast-feed exclusively during the first 6 months and wean thereafter. The nutritional status of children was measured up to age 24 months, using z-scores. Age-specific differences in length-for-age (L/A), weight-for-age (W/A), and weight-for-length (W/L) were compared stratifying by gender and HIV infection status. Multivariable models examined the mean change in z-scores controlling for breast-feeding and other factors. RESULTS: In this analysis, 1589 children were included. Boys had significantly lower L/A scores and became stunted (z-score -<2 standard deviations) earlier than girls. HIV-infected children had significantly lower mean L/A and W/A z-scores than HIV-uninfected children and became stunted and underweight at an earlier age. In multivariable analysis not being breast-fed and being HIV infected were significantly (P < 0.001) associated with decreases in mean L/A, W/A, and W/L z-scores. CONCLUSIONS: This study shows the impact of infant HIV infection on growth and supports the critical importance of breast-feeding. Mother-to-child transmission of HIV programs should endeavor to preserve breast-feeding and find alternative measures to prevent postnatal HIV transmission.

Patient variability in intraoperative ultrasonographic characteristics of colorectal liver metastases.

OBJECTIVE: To determine the distribution of echogenicity (hypoechoic, isoechoic, or hyperechoic) and predominant intraoperative ultrasonography (IOUS) echogenic appearance of colorectal liver metastasis. The interpatient and intrapatient variability of tumor IOUS echogenicity was assessed. DESIGN: Retrospective review of prospectively collected database. SETTING: Tertiary cancer center. PATIENTS: Between January 1998 and July 2001, 99 patients (194 tumors) underwent hepatic resection for colorectal metastases. MAIN OUTCOME MEASURES: During surgery, IOUS of the liver was performed and the images were digitally recorded. Images were randomly coded, blindly reviewed, and scored for echogenicity and ultrasonographic appearance pattern. RESULTS: The ultrasonographic appearance of the colorectal liver metastasis was hypoechoic in 52.0%, isoechoic in 35.7%, and hyperechoic in 12.3% of cases. Most colorectal liver metastases appeared homogeneous (50.8%). Less commonly, identified lesions were characterized by a target or "bull's-eye" appearance (20%) or contained calcifications (19%). Clinicopathologic characteristics, including patient age and sex, as well as tumor size, number, and location and presence of hepatic steatosis, did not correlate with tumor echogenicity or ultrasonographic appearance pattern (all P > .05). Lesions within patients were more similar in echogenicity than lesions between patients (P < .001). Similarly, intrapatient variability in appearance pattern was significantly less than the variability between patients (P = .002). CONCLUSIONS: The ultrasonographic characteristics of hepatic metastases within patients were more similar than between patients. Such information is important because it suggests that, in patients with more than 1 metastasis, the echogenic appearance of an index lesion may predict the echogenic appearance of additional occult disease.

Gender differences in perinatal HIV acquisition among African infants.

OBJECTIVE: We investigated gender-specific risks of mother-to-child transmission (MTCT) at birth and at 6 to 8 weeks among infants born to HIV-infected African women. DESIGN: Follow-up study of infants enrolled in 2 randomized, phase III, clinical trials to prevent MTCT, conducted in Blantyre, Malawi, in southeast Africa. METHODS: Infants were enrolled at birth and monitored postnatally, and their HIV status was assessed at birth and at 6 to 8 weeks (assessment beyond 6-8 weeks is ongoing). Statistical analyses were stratified according to gender, and comparisons were made with descriptive, univariate, and multivariate statistical tests. MTCT was estimated at birth and at 6 to 8 weeks among infants who were not infected at birth. RESULTS: Overall, 966 boys and 998 girls were enrolled. The rate of HIV transmission at birth was 9.5% (187 of 1964 infants). However, at birth significantly more girls (12.6%) than boys (6.3%) were infected with HIV. This association remained significant after controlling for maternal viral load and other factors. Among infants who were uninfected at birth, 8.7% (135 of 1554 infants) acquired HIV by 6 to 8 weeks; of these infants, more girls acquired HIV (10.0%), compared with boys (7.4%). CONCLUSIONS: Female infants may be more susceptible to HIV infection before birth and continuing after birth. Alternatively, in utero mortality rates of HIV-infected male infants may be disproportionately higher and thus more HIV-infected female infants are born. In areas of sub-Saharan Africa, where HIV infection rates are high among women of reproductive age, the magnitude of the gender transmission differences observed in this study could have clinical, preventive, and demographic implications.