Duplex specific nuclease-amplified detection of microRNA using compact quantum dot-DNA conjugates

Advances in nanotechnology have provided new opportunities for the design of next-generation nucleic acid biosensors and diagnostics. Indeed, combining advances in functional nanoparticles, DNA nanotechnology, and nuclease-enzyme-based amplification can give rise to new assays with advantageous properties. In this work, we developed a microRNA (miRNA) assay using bright fluorescent quantum dots (QDs), simple DNA probes, and the enzyme duplex-specific nuclease. We employed an isothermal target-recycling mechanism, where a single miRNA target triggers the cleavage of many DNA signal probes. The incorporation of DNA-functionalized QDs enabled a quantitative fluorescent readout, mediated by Förster resonance energy transfer (FRET)-based interaction with the DNA signal probes. Our approach splits the reaction in two, performing the enzyme-mediated amplification and QD-based detection steps separately such that each reaction could be optimized for performance of the active components. Target recycling gave ca. 3 orders of magnitude amplification, yielding highly sensitive detection with a limit of 42 fM (or 1.2 amol) of miR-148, with excellent selectivity versus mismatched sequences and other miRNAs. Furthermore, we used an alternative target (miR-21) and FRET pair for direct and absolute quantification of miR-21 in RNA extracts from human cancer and normal cell lines

Correlation of Adherence to the 2012 Infectious Diseases Society of America Practice Guidelines with Patient Outcomes in the Treatment of Diabetic Foot Infections in an Outpatient Parenteral Antimicrobial Programme

Aim To evaluate adherence to the 2012 Infectious Diseases Society of America practice guidelines for the management of patients with diabetic foot infections and to determine an association between adherence and clinical outcome. Methods A retrospective chart review was performed to evaluate the management and clinical outcomes of patients with diabetic foot infections treated with outpatient parenteral antimicrobial therapy between 1 January 2011 and 30 June 2012 at Wishard Health Services/Eskenazi Health. Adherence to individual Infectious Diseases Society of America diabetic foot infection treatment guideline recommendations was measured, and then assessed in relation to clinical outcome. Results A total of 57 patients (61% male, mean age 54 years) with moderate to severe diabetic foot infection met the inclusion criteria. None of the treatment courses of these patients adhered to all the Infectious Diseases Society of America guideline recommendations. The recommendations most frequently adhered to were consultation of appropriate multidisciplinary teams (n=54, 94.7%) and performance of diagnostic imaging (n=52, 89.5%). The recommendations least frequently adhered to were diabetic foot wound classification scoring on admission (n=0, 0%), appropriate culture acquisition (n=12, 21.2%), surgical intervention when indicated (n=32, 46.2%) and appropriate empiric antibiotic selection (n=34, 59.7%). Of 56 patients, 52 (92.9%) experienced clinical cure at the end of outpatient parenteral antimicrobial therapy compared with 34 of 53 patients (64%) at 6 months after the completion of therapy. Adherence to individual guidelines was not associated with clinical outcome. Patients who experienced treatment failure were more likely to have severe diabetic foot infection or peripheral neuropathy. Conclusions Adherence to the Infectious Diseases Society of America diabetic foot infection guideline recommendations was found to be suboptimal in the present study. The effect of adhering to individual Infectious Diseases Society of America diabetic foot infection recommendations on clinical outcome needs to be investigated

The Disk and Dark Halo Mass of the Barred Galaxy NGC 4123. I. Observations

The non-circular streaming motions in barred galaxies are sensitive to the mass of the bar and can be used to lift the degeneracy between disk and dark matter halo encountered when fitting axisymmetric rotation curves of disk galaxies. In this paper, we present photometric and kinematic observations of NGC 4123, a barred galaxy of modest size (V_rot = 130 km/sec, L = 0.7 L_*), which reveal strong non-circular motions. The bar has straight dust lanes and an inner Lindblad resonance. The disk of NGC 4123 has no sign of truncation out to 10 scale lengths, and star-forming regions are found well outside R_25. A Fabry-Perot H-alpha velocity field shows velocity jumps of >100 km/sec at the location of the dust lanes within the bar, indicating shocks in the gas flow. VLA observations yield the velocity field of the H I disk. Axisymmetric mass models yield good fits to the rotation curve outside the bar regionfor disk I-band M/L of 2.25 or less, and dark halos with either isothermal or power-law profiles can fit the data well. In a companion paper, we model the full 2-D velocity field, including non-circular motions, to determine the stellar M/L and the mass of the dark halo.Comment: accepted by ApJ, 16 pages, 9 figures (1 color), uses emulateapj.sty, onecolfloat.st

The Use of a Vaginal Conductivity Probe to Influence Calf Sex Ratio via Altered Insemination Time

One hundred eighty-nine mixed breed beef heifers from 13 consignors enrolled in the MACEP heifer development project were utilized in this study. Heifers were synchronized by feeding 0.5 mg melengestrol acetate (MGA) per head per day for 14 days followed by an injection of prostaglandin F2a (PGF2a; 25 mg Lutalyse®) 17 days after the last MGA feeding. Each heifer was fitted with a Heatwatch® transmitter on the morning of PGF2a administration to facilitate detection of estrus. Vaginal conductivity measurements were taken using an Ovatec® probe every 12 hours for 96 hours beginning at the time of PGF2a injection. Heifers randomly assigned to produce a female calf were inseminated near the onset of estrus (as indicated by probe values of £ 55 on the decline). Heifers randomly assigned to produce a male calf were inseminated approximately 24 hours after the onset of estrus (as indicated by probe values of ³ 60 on the incline). All heifers not inseminated by 96 hours after PGF2a were mass inseminated in an attempt to impregnate as many heifers as possible. Heifers that were diagnosed as pregnant as a result of the artificial insemination were subjected to ultrasonography for fetal sex determination. Only 70 of the 189 heifers (37.0%) exhibited estrus according to Heatwatch® and incidence of estrus was influenced by heifer average daily gain, reproductive tract score, and disposition score. Heifers receiving a disposition score of 3 (78.7) had a higher (P\u3c.05) probe reading at AI than those receiving a disposition score of 1 or 2 (70.8 and 72.5, respectively). Heifers with probe readings at insemination of 80 - 84 and \u3e 84 had lower (P\u3c.05) pregnancy rates to AI (13.6 and 0.0%, respectively) than heifers with probe readings in the ranges of \u3c 60, 60 - 64, 65 - 69, 70 - 74, and 75 - 79 (35.7, 40.9, 31.4, 35.3, and 26.9% respectively). Heifers that were bred when probe values were increasing had a lower (P\u3c.05) percentage of male fetuses (34.4%) than those bred during a period of decreasing probe values (69.2% male fetuses). These results demonstrate that a vaginal conductivity probe may be a useful tool to determine an insemination time that could potentially alter calf sex ratio

Physical Structure and Nature of Supernova Remnants in M101

Supernova remnant (SNR) candidates in the giant spiral galaxy M101 have been previously identified from ground-based H-alpha and [SII] images. We have used archival Hubble Space Telescope (HST) H-alpha and broad-band images as well as stellar photometry of 55 SNR candidates to examine their physical structure, interstellar environment, and underlying stellar population. We have also obtained high-dispersion echelle spectra to search for shocked high-velocity gas in 18 SNR candidates, and identified X-ray counterparts to SNR candidates using data from archival observations made by the Chandra X-ray Observatory. Twenty-one of these 55 SNR candidates studied have X-ray counterparts, although one of them is a known ultra-luminous X-ray source. The multi-wavelength information has been used to assess the nature of each SNR candidate. We find that within this limited sample, ~16% are likely remnants of Type Ia SNe and ~45% are remnants of core-collapse SNe. In addition, about ~36% are large candidates which we suggest are either superbubbles or OB/HII complexes. Existing radio observations are not sensitive enough to detect the non-thermal emission from these SNR candidates. Several radio sources are coincident with X-ray sources, but they are associated with either giant HII regions in M101 or background galaxies. The archival HST H-alpha images do not cover the entire galaxy and thus prevents a complete study of M101. Furthermore, the lack of HST [SII] images precludes searches for small SNR candidates which could not be identified by ground-based observations. Such high-resolution images are needed in order to obtain a complete census of SNRs in M101 for a comprehensive investigation of the distribution, population, and rates of SNe in this galaxy.Comment: 37 pages, 4 Tables, 7 Figures, accepted for publication in the Astronomical Journa

Comparative effectiveness of dual-action versus single-action antidepressants for the treatment of depression in people living with HIV/AIDS

Background Depression is the most common psychiatric comorbidity among people living with HIV/AIDS (PLWHA). Little is known about the comparative effectiveness between different types of antidepressants used to treat depression in this population. We compared the effectiveness of dual-action and single-action antidepressants in PLWHA for achieving remission from depression. Methods We used data from the Centers for AIDS Research Network of Integrated Clinic Systems to identify 1175 new user dual-action or single-action antidepressant treatment episodes occurring from 2005 to 2014 for PLWHA diagnosed with depression. The primary outcome was remission from depression defined as a Patient Health Questionnaire-9 (PHQ-9) score <5. Mean difference in PHQ-9 depressive symptom severity was a secondary outcome. The main approach was an intent-to-treat (ITT) evaluation complemented with a per protocol (PP) sensitivity analysis. Generalized linear models were fitted to estimate treatment effects. Results In ITT analysis, 32% of the episodes ended in remission for both dual-action and single-action antidepressants. The odds ratio (OR) of remission was 1.02 (95%CI=0.63,1.67). In PP analysis, 40% of dual-action episodes ended in remission compared to 32% in single-action episodes. Dual-action episodes had 1.33 times the odds of remission (95%CI=0.55,3.21), however the result was not statistically significant. Non-significant differences were also observed for depressive symptom severity. Limitations Missing data was common but was addressed with inverse probability weights. Conclusions Results suggest that single-action and dual-action antidepressants are equally effective in PLWHA. Remission was uncommon highlighting the need to identify health service delivery strategies that aid HIV providers in achieving full remission of their patients’ depression

Civic Engagement Among Orphans and Non-Orphans in Five Low- and Middle-Income Countries

Background: Communities and nations seeking to foster social responsibility in their youth are interested in understanding factors that predict and promote youth involvement in public activities. Orphans and separated children (OSC) are a vulnerable population whose numbers are increasing, particularly in resource-poor settings. Understanding whether and how OSC are engaged in civic activities is important for community and world leaders who need to provide care for OSC and ensure their involvement in sustainable development. Methods: The Positive Outcomes for Orphans study (POFO) is a multi-country, longitudinal cohort study of OSC randomly sampled from institution-based care and from family-based care, and of non-OSC sampled from the same study regions. Participants represent six sites in five low-and middle-income countries. We examined civic engagement activities and government trust among subjects \u3e =16 years old at 90-month follow-up (approximately 7.5 years after baseline). We calculated prevalences and estimated the association between key demographic variables and prevalence of regular volunteer work using multivariable Poisson regression, with sampling weights to accounting for the complex sampling design. Results: Among the 1,281 POFO participants \u3e =16 who were assessed at 90-month follow-up, 45 % participated in regular community service or volunteer work; two-thirds of those volunteers did so on a strictly voluntary basis. While government trust was fairly high, at approximately 70 % for each level of government, participation in voting was only 15 % among those who were \u3e =18 years old. We did not observe significant associations between demographic characteristics and regular volunteer work, with the exception of large variation by study site. Conclusion: As the world’s leaders grapple with the many competing demands of global health, economic security, and governmental stability, the participation of today’s youth in community and governance is essential for sustainability. This study provides a first step in understanding the degree to which OSC from different care settings across multiple low- and middle-income countries are engaged in their communities

Glycosylated superparamagnetic nanoparticle gradients for osteochondral tissue engineering

In developmental biology, gradients of bioactive signals direct the formation of structural transitions in tissue that are key to physiological function. Failure to reproduce these native features in an in vitro setting can severely limit the success of bioengineered tissue constructs. In this report, we introduce a facile and rapid platform that uses magnetic field alignment of glycosylated superparamagnetic iron oxide nanoparticles, pre-loaded with growth factors, to pattern biochemical gradients into a range of biomaterial systems. Gradients of bone morphogenetic protein 2 in agarose hydrogels were used to spatially direct the osteogenesis of human mesenchymal stem cells and generate robust osteochondral tissue constructs exhibiting a clear mineral transition from bone to cartilage. Interestingly, the smooth gradients in growth factor concentration gave rise to biologically-relevant, emergent structural features, including a tidemark transition demarcating mineralized and non-mineralized tissue and an osteochondral interface rich in hypertrophic chondrocytes. This platform technology offers great versatility and provides an exciting new opportunity for overcoming a range of interfacial tissue engineering challenges

Multiple Overimputation to Address Missing Data and Measurement Error: Application to HIV Treatment During Pregnancy and Pregnancy Outcomes

Investigations of the association of combination antiretroviral therapy (ART) with pregnancy outcomes often rely on routinely collected clinical data, which are prone to missing data and measurement error. Measurement error in gestational age may bias the relationship between combination ART and gestational age-based outcomes

A Comparison of the Wellbeing of Orphans and Abandoned Children Ages 6–12 in Institutional and Community-Based Care Settings in 5 Less Wealthy Nations

Background: Leaders are struggling to care for the estimated 143,000,000 orphans and millions more abandoned children worldwide. Global policy makers are advocating that institution-living orphans and abandoned children (OAC) be moved as quickly as possible to a residential family setting and that institutional care be used as a last resort. This analysis tests the hypothesis that institutional care for OAC aged 6-12 is associated with worse health and wellbeing than community residential care using conservative two-tail tests. Methodology: The Positive Outcomes for Orphans (POFO) study employed two-stage random sampling survey methodology in 6 sites across 5 countries to identify 1,357 institution-living and 1,480 community-living OAC ages 6-12, 658 of whom were double-orphans or abandoned by both biological parents. Survey analytic techniques were used to compare cognitive functioning, emotion, behavior, physical health, and growth. Linear mixed-effects models were used to estimate the proportion of variability in child outcomes attributable to the study site, care setting, and child levels and institutional versus community care settings. Conservative analyses limited the community living children to double-orphans or abandoned children. Principal Findings: Health, emotional and cognitive functioning, and physical growth were no worse for institution-living than community-living OAC, and generally better than for community-living OAC cared for by persons other than a biological parent. Differences between study sites explained 2-23% of the total variability in child outcomes, while differences between care settings within sites explained 8-21%. Differences among children within care settings explained 64-87%. After adjusting for sites, age, and gender, institution vs. community-living explained only 0.3-7% of the variability in child outcomes. Conclusion: This study does not support the hypothesis that institutional care is systematically associated with poorer wellbeing than community care for OAC aged 6-12 in those countries facing the greatest OAC burden. Much greater variability among children within care settings was observed than among care settings type. Methodologically rigorous studies must be conducted in those countries facing the new OAC epidemic in order to understand which characteristics of care promote child wellbeing. Such characteristics may transcend the structural definitions of institutions or family homes