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  1. Evaluation of the HIV-1 reverse transcriptase inhibitory properties of ...

    African Journals Online (AJOL)

    remedies in Kenya were screened for activity against the HIV-1 reverse transcriptase.“ ... drugs at present have fallen short of expectation in many ways. ..... effective doses: aromatic polycyclic diones hypericin and pseudhypericin. Proceedings of the National Academy of. Science of the USA. 1988; 85: 5230-. 5234. Tang J ...

  2. Initiation of HIV Reverse Transcription

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    Roland Marquet

    2010-01-01

    Full Text Available Reverse transcription of retroviral genomes into double stranded DNA is a key event for viral replication. The very first stage of HIV reverse transcription, the initiation step, involves viral and cellular partners that are selectively packaged into the viral particle, leading to an RNA/protein complex with very specific structural and functional features, some of which being, in the case of HIV-1, linked to particular isolates. Recent understanding of the tight spatio-temporal regulation of reverse transcription and its importance for viral infectivity further points toward reverse transcription and potentially its initiation step as an important drug target.

  3. Evaluation of 4-thiazolidinone derivatives as potential reverse transcriptase inhibitors against HIV-1 drug resistant strains.

    Science.gov (United States)

    Suryawanshi, Rahul; Jadhav, Sushama; Makwana, Nandini; Desai, Dipen; Chaturbhuj, Devidas; Sonawani, Archana; Idicula-Thomas, Susan; Murugesan, Vanangamudi; Katti, Seturam B; Tripathy, Srikanth; Paranjape, Ramesh; Kulkarni, Smita

    2017-04-01

    Rapid emergence of drug resistance is crucial in management of HIV infection limiting implementation of efficacious drugs in the ART regimen. Designing new molecules against HIV drug resistant strains is utmost essential. Based on the anti-HIV-1 activity, we selected four 4-thiazolidinone derivatives (S009-1908, S009-1909, S009-1911, S009-1912) and studied their interaction with reverse transcriptase (RT) from a panel of 10 clinical isolates (8 nevirapine resistant and two susceptible) using in silico methods, and inhibition pattern using in vitro cell based assays. On the basis of binding affinity observed in in silico analysis, 2-(2-chloro-6-nitrophenyl)-3-(4, 6-dimethylpyridin-2-yl) thiazolidin-4-one (S009-1912) was identified as the lead molecule followed by S009-1908, S009-1909 and S009-1911. The in vitro activity against the same panel was assessed using TZM-bl assay (IC50: 0.4-11.44µg/ml, TI: 4-126) and subsequently in PBMC assay against a nevirapine resistant clinical isolate (IC50: 0.8-6.65µg/ml, TI: 8.31-11.43) and standard strain from NIH ARRRP (IC50: 0.95-3.6µg/ml, TI: 9-26). The study shows analogue with pyrimidin-2-yl amino substitution at N-3 position of thiazolidin-4-one ring (S009-1908, S009-1909, S009-1911) exhibited enhanced activity as compared to pyridin-2-yl substituted derivatives (S009-1912), suggesting the use 4-thiazolidinones for developing potent inhibitors against HIV-1 drug resistant strains. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. RT-SHIV, an infectious CCR5-tropic chimeric virus suitable for evaluating HIV reverse transcriptase inhibitors in macaque models

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    Emau Peter

    2009-11-01

    Full Text Available Abstract Background Non-nucleoside reverse transcriptase inhibitors (NNRTIs are an important category of drugs for both chemotherapy and prevention of human immunodeficiency virus type 1 (HIV-1 infection. However, current non-human primate (NHP models utilizing simian immunodeficiency virus (SIV or commonly used chimeric SHIV (SIV expressing HIV-1 envelope are inadequate due to the insensitivity to NNRTIs. To develop a NHP model for evaluation of NNRTI compounds, we characterized a RT-SHIV virus that was assembled by replacing the SIVmac239 reverse transcriptase (RT with that of HIV-1HXB2. Since RT-SHIV exhibited in vitro characteristics of high infectivity, CCR5-usage, and sensitivity to HIV-1 specific NNRTIs, this virus was thought to be suitable for mucosal transmission and then was used to carry out a vaginal transmission study in pigtail macaques (Macaca nemestrina. Results RT-SHIV exhibited in vitro characteristics of an infectious CCR5-tropic chimeric virus. This virus was not only highly sensitive to HIV-1 RT specific NNRTIs; its replication was also inhibited by a variety of NRTIs and protease inhibitors. For in vivo vaginal transmission studies, macaques were either pretreated with a single dose of DMPA (depot medroxyprogesterone acetate or left untreated before intravaginal inoculation with 500 or 1,000 TCID50 of RT-SHIV. All macaques became systemically infected by 2 or 3 weeks post-inoculation exhibiting persistent high viremia, marked CD4+T cell depletion, and antiviral antibody response. DMPA-pretreated macaques showed a higher mean plasma viral load after the acute infection stage, highly variable antiviral antibody response, and a higher incidence of AIDS-like disease as compared with macaques without DMPA pretreatment. Conclusion This chimeric RT-SHIV has exhibited productive replication in both macaque and human PBMCs, predominantly CCR5-coreceptor usage for viral entry, and sensitivity to NNRTIs as well as other anti-HIV

  5. Synthesis, biological evaluation and molecular modeling of 2-Hydroxyisoquinoline-1,3-dione analogues as inhibitors of HIV reverse transcriptase associated ribonuclease H and polymerase.

    Science.gov (United States)

    Tang, Jing; Vernekar, Sanjeev Kumar V; Chen, Yue-Lei; Miller, Lena; Huber, Andrew D; Myshakina, Nataliya; Sarafianos, Stefan G; Parniak, Michael A; Wang, Zhengqiang

    2017-06-16

    Human immunodeficiency virus (HIV) reverse transcriptase (RT) associated ribonuclease H (RNase H) remains the only virally encoded enzymatic function not clinically validated as an antiviral target. 2-Hydroxyisoquinoline-1,3-dione (HID) is known to confer active site directed inhibition of divalent metal-dependent enzymatic functions, such as HIV RNase H, integrase (IN) and hepatitis C virus (HCV) NS5B polymerase. We report herein the synthesis and biochemical evaluation of a few C-5, C-6 or C-7 substituted HID subtypes as HIV RNase H inhibitors. Our data indicate that while some of these subtypes inhibited both the RNase H and polymerase (pol) functions of RT, potent and selective RNase H inhibition was achieved with subtypes 8-9 as exemplified with compounds 8c and 9c. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  6. Design, synthesis and biological evaluations of N-Hydroxy thienopyrimidine-2,4-diones as inhibitors of HIV reverse transcriptase-associated RNase H.

    Science.gov (United States)

    Kankanala, Jayakanth; Kirby, Karen A; Huber, Andrew D; Casey, Mary C; Wilson, Daniel J; Sarafianos, Stefan G; Wang, Zhengqiang

    2017-12-01

    Human immunodeficiency virus (HIV) reverse transcriptase (RT) associated ribonuclease H (RNase H) is the only HIV enzymatic function not targeted by current antiviral drugs. Although various chemotypes have been reported to inhibit HIV RNase H, few have shown significant antiviral activities. We report herein the design, synthesis and biological evaluation of a novel N-hydroxy thienopyrimidine-2,3-dione chemotype (11) which potently and selectively inhibited RNase H with considerable potency against HIV-1 in cell culture. Current structure-activity-relationship (SAR) identified analogue 11d as a nanomolar inhibitor of RNase H (IC 50  = 0.04 μM) with decent antiviral potency (EC 50  = 7.4 μM) and no cytotoxicity (CC 50  > 100 μM). In extended biochemical assays compound 11d did not inhibit RT polymerase (pol) while inhibiting integrase strand transfer (INST) with 53 fold lower potency (IC 50  = 2.1 μM) than RNase H inhibition. Crystallographic and molecular modeling studies confirmed the RNase H active site binding mode. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  7. Reverse transcription of the HIV-1 pandemic.

    Science.gov (United States)

    Basavapathruni, Aravind; Anderson, Karen S

    2007-12-01

    The HIV/AIDS pandemic has existed for >25 years. Extensive work globally has provided avenues to combat viral infection, but the disease continues to rage on in the human population and infected approximately 4 million people in 2006 alone. In this review, we provide a brief history of HIV/AIDS, followed by analysis of one therapeutic target of HIV-1: its reverse transcriptase (RT). We discuss the biochemical characterization of RT in order to place emphasis on possible avenues of inhibition, which now includes both nucleoside and non-nucleoside modalities. Therapies against RT remain a cornerstone of anti-HIV treatment, but the virus eventually resists inhibition through the selection of drug-resistant RT mutations. Current inhibitors and associated resistance are discussed, with the hopes that new therapeutics can be developed against RT.

  8. Natural Plant Alkaloid (Emetine Inhibits HIV-1 Replication by Interfering with Reverse Transcriptase Activity

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    Ana Luiza Chaves Valadão

    2015-06-01

    Full Text Available Ipecac alkaloids are secondary metabolites produced in the medicinal plant Psychotria ipecacuanha. Emetine is the main alkaloid of ipecac and one of the active compounds in syrup of Ipecac with emetic property. Here we evaluated emetine’s potential as an antiviral agent against Human Immunodeficiency Virus. We performed in vitro Reverse Transcriptase (RT Assay and Natural Endogenous Reverse Transcriptase Activity Assay (NERT to evaluate HIV RT inhibition. Emetine molecular docking on HIV-1 RT was also analyzed. Phenotypic assays were performed in non-lymphocytic and in Peripheral Blood Mononuclear Cells (PBMC with HIV-1 wild-type and HIV-harboring RT-resistant mutation to Nucleoside Reverse Transcriptase Inhibitors (M184V. Our results showed that HIV-1 RT was blocked in the presence of emetine in both models: in vitro reactions with isolated HIV-1 RT and intravirion, measured by NERT. Emetine revealed a strong potential of inhibiting HIV-1 replication in both cellular models, reaching 80% of reduction in HIV-1 infection, with low cytotoxic effect. Emetine also blocked HIV-1 infection of RT M184V mutant. These results suggest that emetine is able to penetrate in intact HIV particles, and bind and block reverse transcription reaction, suggesting that it can be used as anti-HIV microbicide. Taken together, our findings provide additional pharmacological information on the potential therapeutic effects of emetine.

  9. Plasma drug concentrations and virologic evaluations after stopping treatment with nonnucleoside reverse-transcriptase inhibitors in HIV type 1-infected children.

    NARCIS (Netherlands)

    Cressey, T.R.; Green, H.; Khoo, S.; Treluyer, J.M.; Compagnucci, A.; Saidi, Y.; Lallemant, M.; Gibb, D.M.; Burger, D.M.

    2008-01-01

    BACKGROUND: The optimum strategy for stopping treatment with drugs that have different half-lives in a combination regimen to minimize the risk of selecting drug-resistant viruses remains unknown. We evaluated drug concentrations in plasma, human immunodeficiency virus (HIV) load, and development of

  10. HIV-1 Non-Nucleoside Reverse Transcriptase Inhibitors

    DEFF Research Database (Denmark)

    Vanangamudi, Murugesan; Poongavanam, Vasanthanathan; Namasivayam, Vigneshwaran

    2017-01-01

    BACKGROUND: Design of inhibitors for HIV-1 reverse transcriptase inhibition (HIV-1 RT) is one of the successful chemotherapies for the treatment of HIV infection. Among the inhibitors available for HIV-1 RT, non-nucleoside reverse transcriptase inhibitors (NNRTIs) have shown to be very promising...... and clinically approved drugs. However, the efficiency of many of these drugs has been reduced by the drug-resistant variants of HIV-1 RT. The aim of the current review is to provide a summary of lead optimization strategies from the 3D-QSARs studies on NNRTI class from the past 21 years (1995 to 2016). METHODS......, formimidoester disulfides, thiocarbamate, thiazolidinone derivatives, etc. have been discussed in detail. In addition, we explore the position of the functional groups that drive the protein-ligand interaction. RESULTS: The structure-activity relationship (SAR) revealed from CoMFA and CoMSIA studies...

  11. Reverse Transcription Mechanically Initiates HIV-1 Capsid Disassembly.

    Science.gov (United States)

    Rankovic, Sanela; Varadarajan, Janani; Ramalho, Ruben; Aiken, Christopher; Rousso, Itay

    2017-06-15

    The HIV-1 core consists of the viral genomic RNA and several viral proteins encased within a conical capsid. After cell entry, the core disassembles in a process termed uncoating. Although HIV-1 uncoating has been linked to reverse transcription of the viral genome in target cells, the mechanism by which uncoating is initiated is unknown. Using time-lapse atomic force microscopy, we analyzed the morphology and physical properties of isolated HIV-1 cores during the course of reverse transcription in vitro We found that, during an early stage of reverse transcription the pressure inside the capsid increases, reaching a maximum after 7 h. High-resolution mechanical mapping reveals the formation of a stiff coiled filamentous structure underneath the capsid surface. Subsequently, this coiled structure disappears, the stiffness of the capsid drops precipitously to a value below that of a pre-reverse transcription core, and the capsid undergoes partial or complete rupture near the narrow end of the conical structure. We propose that the transcription of the relatively flexible single-stranded RNA into a more rigid filamentous structure elevates the pressure within the core, which triggers the initiation of capsid disassembly.IMPORTANCE For successful infection, the HIV-1 genome, which is in the form of a single-stranded RNA enclosed inside a capsid shell, must be reverse transcribed into double-stranded DNA and released from the capsid (in a process known as uncoating) before it can be integrated into the target cell genome. The mechanism that triggers uncoating is a pivotal question of long standing. By using atomic force microscopy, we found that during reverse transcription the pressure inside the capsid increases until the internal stress exceeds the strength of the capsid structure and the capsid breaks open. The application of AFM technologies to study purified HIV-1 cores represents a new experimental platform for elucidating additional aspects of capsid

  12. Development and evaluation of an assay for HIV-1 protease and reverse transcriptase drug resistance genotyping of all major group-M subtypes

    NARCIS (Netherlands)

    Aitken, S.C.; Kliphuis, A.; Wallis, C.L.; Chu, M.L.; Fillekes, Q.; Barth, R.; Stevens, W.B.; Rinke de Wit, T.F.; Schuurman, R.

    2012-01-01

    BACKGROUND: High cost and varying sensitivity for non-B HIV-1 subtypes limits application of current commercial kits for HIV-1 drug resistance genotyping of all major HIV-1 group-M subtypes. OBJECTIVES: Our research aimed to develop and validate an assay specific for all major HIV-1 group-M subtypes

  13. Interaction between Reverse Transcriptase and Integrase Is Required for Reverse Transcription during HIV-1 Replication.

    Science.gov (United States)

    Tekeste, Shewit S; Wilkinson, Thomas A; Weiner, Ethan M; Xu, Xiaowen; Miller, Jennifer T; Le Grice, Stuart F J; Clubb, Robert T; Chow, Samson A

    2015-12-01

    Human immunodeficiency virus type 1 (HIV-1) replication requires reverse transcription of its RNA genome into a double-stranded cDNA copy, which is then integrated into the host cell chromosome. The essential steps of reverse transcription and integration are catalyzed by the viral enzymes reverse transcriptase (RT) and integrase (IN), respectively. In vitro, HIV-1 RT can bind with IN, and the C-terminal domain (CTD) of IN is necessary and sufficient for this binding. To better define the RT-IN interaction, we performed nuclear magnetic resonance (NMR) spectroscopy experiments to map a binding surface on the IN CTD in the presence of RT prebound to a duplex DNA construct that mimics the primer-binding site in the HIV-1 genome. To determine the biological significance of the RT-IN interaction during viral replication, we used the NMR chemical shift mapping information as a guide to introduce single amino acid substitutions of nine different residues on the putative RT-binding surface in the IN CTD. We found that six viral clones bearing such IN substitutions (R231E, W243E, G247E, A248E, V250E, and I251E) were noninfectious. Further analyses of the replication-defective IN mutants indicated that the block in replication took place specifically during early reverse transcription. The recombinant INs purified from these mutants, though retaining enzymatic activities, had diminished ability to bind RT in a cosedimentation assay. The results indicate that the RT-IN interaction is functionally relevant during the reverse transcription step of the HIV-1 life cycle. To establish a productive infection, human immunodeficiency virus type 1 (HIV-1) needs to reverse transcribe its RNA genome to create a double-stranded DNA copy and then integrate this viral DNA genome into the chromosome of the host cell. These two essential steps are catalyzed by the HIV-1 enzymes reverse transcriptase (RT) and integrase (IN), respectively. We have shown previously that IN physically interacts

  14. APOBEC3G inhibits elongation of HIV-1 reverse transcripts.

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    Kate N Bishop

    2008-12-01

    Full Text Available APOBEC3G (A3G is a host cytidine deaminase that, in the absence of Vif, restricts HIV-1 replication and reduces the amount of viral DNA that accumulates in cells. Initial studies determined that A3G induces extensive mutation of nascent HIV-1 cDNA during reverse transcription. It has been proposed that this triggers the degradation of the viral DNA, but there is now mounting evidence that this mechanism may not be correct. Here, we use a natural endogenous reverse transcriptase assay to show that, in cell-free virus particles, A3G is able to inhibit HIV-1 cDNA accumulation not only in the absence of hypermutation but also without the apparent need for any target cell factors. We find that although reverse transcription initiates in the presence of A3G, elongation of the cDNA product is impeded. These data support the model that A3G reduces HIV-1 cDNA levels by inhibiting synthesis rather than by inducing degradation.

  15. Drug Resistance in Non-B Subtype HIV-1: Impact of HIV-1 Reverse Transcriptase Inhibitors

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    Kamalendra Singh

    2014-09-01

    Full Text Available Human immunodeficiency virus (HIV causes approximately 2.5 million new infections every year, and nearly 1.6 million patients succumb to HIV each year. Several factors, including cross-species transmission and error-prone replication have resulted in extraordinary genetic diversity of HIV groups. One of these groups, known as group M (main contains nine subtypes (A-D, F-H and J-K and causes ~95% of all HIV infections. Most reported data on susceptibility and resistance to anti-HIV therapies are from subtype B HIV infections, which are prevalent in developed countries but account for only ~12% of all global HIV infections, whereas non-B subtype HIV infections that account for ~88% of all HIV infections are prevalent primarily in low and middle-income countries. Although the treatments for subtype B infections are generally effective against non-B subtype infections, there are differences in response to therapies. Here, we review how polymorphisms, transmission efficiency of drug-resistant strains, and differences in genetic barrier for drug resistance can differentially alter the response to reverse transcriptase-targeting therapies in various subtypes.

  16. Inhibition of Reverse Transcriptase Activity Increases Stability of the HIV-1 Core

    Science.gov (United States)

    Yang, Yang; Fricke, Thomas

    2013-01-01

    Previous studies showed that HIV-1 reverse transcription occurs during or before uncoating, linking mechanistically reverse transcription with uncoating. Here we show that inhibition of reverse transcriptase (RT) during HIV-1 infection by pharmacologic or genetic means increased the stability of the HIV-1 core during infection. Interestingly, HIV-1 particles with increased core stability were resistant to the core-destabilizing effects of rhesus TRIM5α (TRIM5αrh). Collectively, this work implies that the surface of the HIV-1 core is dynamic and changes upon the ongoing processes within the core. PMID:23077298

  17. NMR structure of the HIV-1 reverse transcriptase thumb subdomain

    Energy Technology Data Exchange (ETDEWEB)

    Sharaf, Naima G. [University of Pittsburgh, School of Medicine, Department of Structural Biology and Pittsburgh Center for HIV Protein Interactions (United States); Brereton, Andrew E. [Oregon State University, Department of Biochemistry and Biophysics, 2011 Ag & Life Sciences Bldg (United States); Byeon, In-Ja L. [University of Pittsburgh, School of Medicine, Department of Structural Biology and Pittsburgh Center for HIV Protein Interactions (United States); Andrew Karplus, P. [Oregon State University, Department of Biochemistry and Biophysics, 2011 Ag & Life Sciences Bldg (United States); Gronenborn, Angela M., E-mail: amg100@pitt.edu [University of Pittsburgh, School of Medicine, Department of Structural Biology and Pittsburgh Center for HIV Protein Interactions (United States)

    2016-12-15

    The solution NMR structure of the isolated thumb subdomain of HIV-1 reverse transcriptase (RT) has been determined. A detailed comparison of the current structure with dozens of the highest resolution crystal structures of this domain in the context of the full-length enzyme reveals that the overall structures are very similar, with only two regions exhibiting local conformational differences. The C-terminal capping pattern of the αH helix is subtly different, and the loop connecting the αI and αJ helices in the p51 chain of the full-length p51/p66 heterodimeric RT differs from our NMR structure due to unique packing interactions in mature RT. Overall, our data show that the thumb subdomain folds independently and essentially the same in isolation as in its natural structural context.

  18. Evaluation of three enzyme immunoassays for HIV-1 antigen detection.

    Science.gov (United States)

    Willoughby, P B; Lisker, A; Folds, J D

    1989-01-01

    Three enzyme immunoassay (EIA) methods for the detection of human immunodeficiency virus (HIV-1) were evaluated. Serum or plasma samples from 22 individuals seropositive for HIV-1 antibodies were tested with the Abbott, Coulter, and DuPont kits for presence of HIV-1 p24 antigen. Another 12 samples were tested with two kits only. Discordant results were obtained with 9 of 34 (26%) HIV-1-antibody-positive patient samples tested. Most of these discrepancies were found in samples containing less than 30 pg/ml of HIV-1 p24 core antigen. A sampling of sera from normal blood donors and patients with infectious or autoimmune diseases revealed a low level of false positive reactions, especially with sera containing antinuclear antibodies or rheumatoid factor. Noteworthy is the frequency of false positive reactions seen with the DuPont EIA for HIV-1 p24 antigen. 18/111 sera (16.2%) containing auto-antibodies tested positively with the DuPont HIV-1 p24 antigen EIA. The nonspecific nature of the test reactivity for 9/10 of these samples was confirmed using an HIV-1 p24 antigen inhibition assay. These findings are discussed in light of the need for HIV-1 antigen detection in the clinical laboratory and of other methods for HIV-1 detection: the polymerase chain reaction and measurements of reverse transcriptase activity.

  19. HIV-1 Protease, Reverse Transcriptase, and Integrase Variation.

    Science.gov (United States)

    Rhee, Soo-Yon; Sankaran, Kris; Varghese, Vici; Winters, Mark A; Hurt, Christopher B; Eron, Joseph J; Parkin, Neil; Holmes, Susan P; Holodniy, Mark; Shafer, Robert W

    2016-07-01

    HIV-1 protease (PR), reverse transcriptase (RT), and integrase (IN) variability presents a challenge to laboratories performing genotypic resistance testing. This challenge will grow with increased sequencing of samples enriched for proviral DNA such as dried blood spots and increased use of next-generation sequencing (NGS) to detect low-abundance HIV-1 variants. We analyzed PR and RT sequences from >100,000 individuals and IN sequences from >10,000 individuals to characterize variation at each amino acid position, identify mutations indicating APOBEC-mediated G-to-A editing, and identify mutations resulting from selective drug pressure. Forty-seven percent of PR, 37% of RT, and 34% of IN positions had one or more amino acid variants with a prevalence of ≥1%. Seventy percent of PR, 60% of RT, and 60% of IN positions had one or more variants with a prevalence of ≥0.1%. Overall 201 PR, 636 RT, and 346 IN variants had a prevalence of ≥0.1%. The median intersubtype prevalence ratios were 2.9-, 2.1-, and 1.9-fold for these PR, RT, and IN variants, respectively. Only 5.0% of PR, 3.7% of RT, and 2.0% of IN variants had a median intersubtype prevalence ratio of ≥10-fold. Variants at lower prevalences were more likely to differ biochemically and to be part of an electrophoretic mixture compared to high-prevalence variants. There were 209 mutations indicative of APOBEC-mediated G-to-A editing and 326 mutations nonpolymorphic treatment selected. Identification of viruses with a high number of APOBEC-associated mutations will facilitate the quality control of dried blood spot sequencing. Identifying sequences with a high proportion of rare mutations will facilitate the quality control of NGS. Most antiretroviral drugs target three HIV-1 proteins: PR, RT, and IN. These proteins are highly variable: many different amino acids can be present at the same position in viruses from different individuals. Some of the amino acid variants cause drug resistance and occur mainly

  20. Inhibitory effect of aqueous dandelion extract on HIV-1 replication and reverse transcriptase activity

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    Han Huamin

    2011-11-01

    Full Text Available Abstract Background Acquired immunodeficiency syndrome (AIDS, which is caused by the human immunodeficiency virus (HIV, is an immunosuppressive disease that results in life-threatening opportunistic infections. The general problems in current therapy include the constant emergence of drug-resistant HIV strains, adverse side effects and the unavailability of treatments in developing countries. Natural products from herbs with the abilities to inhibit HIV-1 life cycle at different stages, have served as excellent sources of new anti-HIV-1 drugs. In this study, we aimed to investigate the anti-HIV-1 activity of aqueous dandelion extract. Methods The pseudotyped HIV-1 virus has been utilized to explore the anti-HIV-1 activity of dandelion, the level of HIV-1 replication was assessed by the percentage of GFP-positive cells. The inhibitory effect of the dandelion extract on reverse transcriptase activity was assessed by the reverse transcriptase assay kit. Results Compared to control values obtained from cells infected without treatment, the level of HIV-1 replication and reverse transcriptase activity were decreased in a dose-dependent manner. The data suggest that dandelion extract has a potent inhibitory activity against HIV-1 replication and reverse transcriptase activity. The identification of HIV-1 antiviral compounds from Taraxacum officinale should be pursued. Conclusions The dandelion extract showed strong activity against HIV-1 RT and inhibited both the HIV-1 vector and the hybrid-MoMuLV/MoMuSV retrovirus replication. These findings provide additional support for the potential therapeutic efficacy of Taraxacum officinale. Extracts from this plant may be regarded as another starting point for the development of an antiretroviral therapy with fewer side effects.

  1. Inhibitory effect of aqueous dandelion extract on HIV-1 replication and reverse transcriptase activity

    Science.gov (United States)

    2011-01-01

    Background Acquired immunodeficiency syndrome (AIDS), which is caused by the human immunodeficiency virus (HIV), is an immunosuppressive disease that results in life-threatening opportunistic infections. The general problems in current therapy include the constant emergence of drug-resistant HIV strains, adverse side effects and the unavailability of treatments in developing countries. Natural products from herbs with the abilities to inhibit HIV-1 life cycle at different stages, have served as excellent sources of new anti-HIV-1 drugs. In this study, we aimed to investigate the anti-HIV-1 activity of aqueous dandelion extract. Methods The pseudotyped HIV-1 virus has been utilized to explore the anti-HIV-1 activity of dandelion, the level of HIV-1 replication was assessed by the percentage of GFP-positive cells. The inhibitory effect of the dandelion extract on reverse transcriptase activity was assessed by the reverse transcriptase assay kit. Results Compared to control values obtained from cells infected without treatment, the level of HIV-1 replication and reverse transcriptase activity were decreased in a dose-dependent manner. The data suggest that dandelion extract has a potent inhibitory activity against HIV-1 replication and reverse transcriptase activity. The identification of HIV-1 antiviral compounds from Taraxacum officinale should be pursued. Conclusions The dandelion extract showed strong activity against HIV-1 RT and inhibited both the HIV-1 vector and the hybrid-MoMuLV/MoMuSV retrovirus replication. These findings provide additional support for the potential therapeutic efficacy of Taraxacum officinale. Extracts from this plant may be regarded as another starting point for the development of an antiretroviral therapy with fewer side effects. PMID:22078030

  2. Focus on Chirality of HIV-1 Non-Nucleoside Reverse Transcriptase Inhibitors

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    Valeria Famiglini

    2016-02-01

    Full Text Available Chiral HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs are of great interest since one enantiomer is often more potent than the corresponding counterpart against the HIV-1 wild type (WT and the HIV-1 drug resistant mutant strains. This review exemplifies the various studies made to investigate the effect of chirality on the antiretroviral activity of top HIV-1 NNRTI compounds, such as nevirapine (NVP, efavirenz (EFV, alkynyl- and alkenylquinazolinone DuPont compounds (DPC, diarylpyrimidine (DAPY, dihydroalkyloxybenzyloxopyrimidine (DABO, phenethylthiazolylthiourea (PETT, indolylarylsulfone (IAS, arylphosphoindole (API and trifluoromethylated indole (TFMI The chiral separation, the enantiosynthesis, along with the biological properties of these HIV-1 NNRTIs, are discussed.

  3. [Process of HIV-1 reverse transcription and its detection by using PCR].

    Science.gov (United States)

    Yao, Wen-Xue; Wu, Ying-Liang; Guo, Ying

    2008-02-01

    Human immunodeficiency virus (HIV) is a retrovirus, belongs to Lentiviridae family. As long as viral genetic material entering into host cytoplasm, double-strand DNAs synthesis occurs which is catalyzed by reverse transcriptase (RT) with viral plus-strand RNA as template. This reverse transcription is a key link of HIV-1 life cycle and an important target for anti-HIV drug development. The process of reverse transcription can be divided into several steps: formation of minus-strand strong-stop DNA; the first translocation; initiation of plus-strand DNA synthesis; and, the second translocation and the completion of both strands. These steps can be detected individually by using polymerase chain reaction (PCR) according to the amplified products on the region of R/U5, U3, U5/PBS and the sequence between LTR and Gag. In this review, we summarize the principle for detecting stages of HIV-1 reverse transcription by using PCR.

  4. Sargassum fusiforme fraction is a potent and specific inhibitor of HIV-1 fusion and reverse transcriptase

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    Thornber Carol

    2008-01-01

    Full Text Available Abstract Sargassum fusiforme (Harvey Setchell has been shown to be a highly effective inhibitor of HIV-1 infection. To identify its mechanism of action, we performed bioactivity-guided fractionation on Sargassum fusiforme mixture. Here, we report isolation of a bioactive fraction SP4-2 (S. fusiforme, which at 8 μg/ml inhibited HIV-1 infection by 86.9%, with IC50 value of 3.7 μg. That represents 230-fold enhancement of antiretroviral potency as compared to the whole extract. Inhibition was mediated against both CXCR4 (X4 and CCR5 (R5 tropic HIV-1. Specifically, 10 μg/ml SP4-2 blocked HIV-1 fusion and entry by 53%. This effect was reversed by interaction of SP4-2 with sCD4, suggesting that S. fusiforme inhibits HIV-1 infection by blocking CD4 receptor, which also explained observed inhibition of both X4 and R5-tropic HIV-1. SP4-2 also inhibited HIV-1 replication after virus entry, by directly inhibiting HIV-1 reverse transcriptase (RT in a dose dependent manner by up to 79%. We conclude that the SP4-2 fraction contains at least two distinct and biologically active molecules, one that inhibits HIV-1 fusion by interacting with CD4 receptor, and another that directly inhibits HIV-1 RT. We propose that S. fusiforme is a lead candidate for anti-HIV-1 drug development.

  5. HIV Nonnucleoside Reverse Transcriptase Inhibitors and Trimethoprim-Sulfamethoxazole Inhibit Plasmodium Liver Stages

    OpenAIRE

    Hobbs, Charlotte V.; Voza, Tatiana; de la Vega, Patricia; Vanvliet, Jillian; Conteh, Solomon; Penzak, Scott R.; Michael P Fay; Anders, Nicole; Ilmet, Tiina; Li, Yonghua; Borkowsky, William; Krzych, Urszula; Duffy, Patrick E.; Sinnis, Photini

    2012-01-01

    Background. Although nonnucleoside reverse transcriptase inhibitors (NNRTIs) are usually part of first-line treatment regimens for human immunodeficiency virus (HIV), their activity on Plasmodium liver stages remains unexplored. Additionally, trimethoprim-sulfamethoxazole (TMP-SMX), used for opportunistic infection prophylaxis in HIV-exposed infants and HIV-infected patients, reduces clinical episodes of malaria; however, TMP-SMX effect on Plasmodium liver stages requires further study.

  6. Ellagic acid & gallic acid from Lagerstroemia speciosa L. inhibit HIV-1 infection through inhibition of HIV-1 protease & reverse transcriptase activity

    Directory of Open Access Journals (Sweden)

    Nutan

    2013-01-01

    Interpretation & conclusions: The present study shows a novel anti-HIV activity of banaba. The active components responsible for anti-HIV activity were gallic acid and ellagic acid, through inhibition of reverse transcriptase and HIV protease, respectively and hence could be regarded as promising candidates for the development of topical anti-HIV-1 agents.

  7. Prevalence of Primary HIV Drug Resistance in Thailand Detected by Short Reverse Transcriptase Genotypic Resistance Assay.

    Science.gov (United States)

    Kiertiburanakul, Sasisopin; Pinsai, Subencha; Chantratita, Wasun; Pasomsub, Ekawat; Leechawengwongs, Manoon; Thipmontree, Wilawan; Siriyakorn, Nirada; Sungkanuparph, Somnuek

    2016-01-01

    HIV drug resistance (HIVDR) is the major cause of treatment failure after scaling up of antiretroviral therapy (ART). HIVDR testing prior to ART initiation is not routinely performed in resource-limited settings. We aimed to assess the prevalence of primary HIVDR by short reverse transcriptase (RT) genotypic resistance assay and evaluate of the impact of the mutations on the treatment outcomes. A prospective cohort study was conducted in treatment-naïve HIV-infected patients. Fourteen major mutations of codon 99-191 on the RT gene were selected (K103N, V106A/M, V108I, Q151M, Y181C/I, M184V/I, Y188C/L/H, and G190S/A) at a cost of testing of 35 USD. The association between the presence of primary HIVDR and undetectable HIV RNA (logistic regression, factors associated with undetectable HIV RNA after 6 months of ART were: having M184V/I (odds ratio [OR] 0.11; 95% confidence interval [CI] 0.02-0.62, p = 0.013), condom use (OR 2.38; 95% CI 1.12-5.06, p = 0.024), and adherence per 5% increase (OR 1.16; 95% CI 1.00-1.35, p = 0.044). The prevalence of primary HIVDR is approximately 8%; it is associated with detectable HIV RNA at 6 months after ART initiation. Routine "short RT" genotypic resistance assay should be considered in resource-limited settings to maximize treatment outcome.

  8. SJ-3366, a unique and highly potent nonnucleoside reverse transcriptase inhibitor of human immunodeficiency virus type 1 (HIV-1) that also inhibits HIV-2.

    Science.gov (United States)

    Buckheit, R W; Watson, K; Fliakas-Boltz, V; Russell, J; Loftus, T L; Osterling, M C; Turpin, J A; Pallansch, L A; White, E L; Lee, J W; Lee, S H; Oh, J W; Kwon, H S; Chung, S G; Cho, E H

    2001-02-01

    We have identified and characterized a potent new nonnucleoside reverse transcriptase (RT) inhibitor (NNRTI) of human immunodeficiency virus type 1 (HIV-1) that also is active against HIV-2 and which interferes with virus replication by two distinct mechanisms. 1-(3-Cyclopenten-1-yl)methyl-6-(3,5-dimethylbenzoyl)-5-ethyl-2,4-pyrimidinedione (SJ-3366) inhibits HIV-1 replication at concentrations of approximately 1 nM, with a therapeutic index of greater than 4 x 10(6). The efficacy and toxicity of SJ-3366 are consistent when evaluated with established or fresh human cells, and the compound is equipotent against all strains of HIV-1 evaluated, including syncytium-inducing, non-syncytium-inducing, monocyte/macrophage-tropic, and subtype virus strains. Distinct from other members of the pharmacologic class of NNRTIs, SJ-3366 inhibited laboratory and clinical strains of HIV-2 at a concentration of approximately 150 nM, yielding a therapeutic index of approximately 20,000. Like most NNRTIs, the compound was less active when challenged with HIV-1 strains possessing the Y181C, K103N, and Y188C amino acid changes in the RT and selected for a virus with a Y181C amino acid change in the RT after five tissue culture passages in the presence of the compound. In combination anti-HIV assays with nucleoside and nonnucleoside RT and protease inhibitors, additive interactions occurred with all compounds tested with the exception of dideoxyinosine, with which a synergistic interaction was found. Biochemically, SJ-3366 exhibited a K(i) value of 3.2 nM, with a mixed mechanism of inhibition against HIV-1 RT, but it did not inhibit HIV-2 RT. SJ-3366 also inhibited the entry of both HIV-1 and HIV-2 into target cells. On the basis of its therapeutic index and multiple mechanisms of anti-HIV action, SJ-3366 represents an exciting new compound for use in HIV-infected individuals.

  9. HIV/AIDS Prevention Program Evaluation Report.

    Science.gov (United States)

    Amaro, Hortensia; Barker, Marybeth; Cassisy, Theresa; Hardy-Fanta, Carol; Hereen, Tim; Levenson, Suzette; McCloskey, Lois; Melendez, Michael

    This report addresses the four research objectives that were established by the Massachusetts Primary Prevention Group (MPPG) and the Massachusetts Department of Public Health's HIV/AIDS Bureau. The objectives were to: (1) review and summarize literature that formally evaluated HIV prevention interventions; (2) describe how currently funded…

  10. Anti-HIV-1 ADCC Antibodies following Latency Reversal and Treatment Interruption.

    Science.gov (United States)

    Lee, Wen Shi; Kristensen, Anne B; Rasmussen, Thomas A; Tolstrup, Martin; Østergaard, Lars; Søgaard, Ole S; Wines, Bruce D; Hogarth, P Mark; Reynaldi, Arnold; Davenport, Miles P; Emery, Sean; Amin, Janaki; Cooper, David A; Kan, Virginia L; Fox, Julie; Gruell, Henning; Parsons, Matthew S; Kent, Stephen J

    2017-08-01

    There is growing interest in utilizing antibody-dependent cellular cytotoxicity (ADCC) to eliminate infected cells following reactivation from HIV-1 latency. A potential barrier is that HIV-1-specific ADCC antibodies decline in patients on long-term antiretroviral therapy (ART) and may not be sufficient to eliminate reactivated latently infected cells. It is not known whether reactivation from latency with latency-reversing agents (LRAs) could provide sufficient antigenic stimulus to boost HIV-1-specific ADCC. We found that treatment with the LRA panobinostat or a short analytical treatment interruption (ATI), 21 to 59 days, was not sufficient to stimulate an increase in ADCC-competent antibodies, despite viral rebound in all subjects who underwent the short ATI. In contrast, a longer ATI, 2 to 12 months, among subjects enrolled in the Strategies for Management of Antiretroviral Therapy (SMART) trial robustly boosted HIV-1 gp120-specific Fc receptor-binding antibodies and ADCC against HIV-1-infected cells in vitro These results show that there is a lag between viral recrudescence and the boosting of ADCC antibodies, which has implications for strategies toward eliminating latently infected cells.IMPORTANCE The "shock and kill" HIV-1 cure strategy aims to reactivate HIV-1 expression in latently infected cells and subsequently eliminate the reactivated cells through immune-mediated killing. Several latency reversing agents (LRAs) have been examined in vivo, but LRAs alone have not been able to achieve HIV-1 remission and prevent viral rebound following analytical treatment interruption (ATI). In this study, we examined whether LRA treatment or ATI can provide sufficient antigenic stimulus to boost HIV-1-specific functional antibodies that can eliminate HIV-1-infected cells. Our study has implications for the antigenic stimulus required for antilatency strategies and/or therapeutic vaccines to boost functional antibodies and assist in eliminating the latent reservoir

  11. Ultrasensitive HIV-1 p24 Assay Detects Single Infected Cells and Differences in Reservoir Induction by Latency Reversal Agents.

    Science.gov (United States)

    Passaes, Caroline Pereira Bittencourt; Bruel, Timothée; Decalf, Jérémie; David, Annie; Angin, Mathieu; Monceaux, Valerie; Muller-Trutwin, Michaela; Noel, Nicolas; Bourdic, Katia; Lambotte, Olivier; Albert, Matthew L; Duffy, Darragh; Schwartz, Olivier; Sáez-Cirión, Asier

    2017-03-15

    The existence of HIV reservoirs in infected individuals under combined antiretroviral therapy (cART) represents a major obstacle toward cure. Viral reservoirs are assessed by quantification of HIV nucleic acids, a method which does not discriminate between infectious and defective viruses, or by viral outgrowth assays, which require large numbers of cells and long-term cultures. Here, we used an ultrasensitive p24 digital assay, which we report to be 1,000-fold more sensitive than classical enzyme-linked immunosorbent assays (ELISAs) in the quantification of HIV-1 Gag p24 production in samples from HIV-infected individuals. Results from ultrasensitive p24 assays were compared to those from conventional viral RNA reverse transcription-quantitative PCR (RT-qPCR)-based assays and from outgrowth assay readout by flow cytometry. Using serial dilutions and flow-based single-cell sorting, we show that viral proteins produced by a single infected cell can be detected by the ultrasensitive p24 assay. This unique sensitivity allowed the early (as soon as day 1 in 43% of cases) and more efficient detection and quantification of p24 in phytohemagglutinin-L (PHA)-stimulated CD4(+) T cells from individuals under effective cART. When seven different classes of latency reversal agents (LRA) in resting CD4(+) T cells from HIV-infected individuals were tested, the ultrasensitive p24 assay revealed differences in the extent of HIV reactivation. Of note, HIV RNA production was infrequently accompanied by p24 protein production (19%). Among the drugs tested, prostratin showed a superior capacity in inducing viral protein production. In summary, the ultrasensitive p24 assay allows the detection and quantification of p24 produced by single infected CD4(+) T cells and provides a unique tool to assess early reactivation of infectious virus from reservoirs in HIV-infected individuals.IMPORTANCE The persistence of HIV reservoirs in infected individuals under effective antiretroviral

  12. JAK-STAT Signaling Pathways and Inhibitors Affect Reversion of Envelope-Mutated HIV-1.

    Science.gov (United States)

    Quan, Yudong; Xu, Hongtao; Han, Yingshan; Mesplède, Thibault; Wainberg, Mark A

    2017-05-01

    HIV can spread by both cell-free and cell-to-cell transmission. Here, we show that many of the amino acid changes in Env that are close to the CD4 binding pocket can affect HIV replication. We generated a number of mutant viruses that were unable to infect T cells as cell-free viruses but were nevertheless able to infect certain T cell lines as cell-associated viruses, which was followed by reversion to the wild type. However, the activation of JAK-STAT signaling pathways caused the inhibition of such cell-to-cell infection as well as the reversion of multiple HIV Env mutants that displayed differences in their abilities to bind to the CD4 receptor. Specifically, two T cell activators, interleukin-2 (IL-2) and phorbol 12-myristate 13-acetate (PMA), both capable of activation of JAK-STAT pathways, were able to inhibit cell-to-cell viral transmission. In contrast, but consistent with the above result, a number of JAK-STAT and mTOR inhibitors actually promoted HIV-1 transmission and reversion. Hence, JAK-STAT signaling pathways may differentially affect the replication of a variety of HIV Env mutants in ways that differ from the role that these pathways play in the replication of wild-type viruses.IMPORTANCE Specific alterations in HIV Env close to the CD4 binding site can differentially change the ability of HIV to mediate infection for cell-free and cell-associated viruses. However, such differences are dependent to some extent on the types of target cells used. JAK-STAT signaling pathways are able to play major roles in these processes. This work sheds new light on factors that can govern HIV infection of target cells. Copyright © 2017 American Society for Microbiology.

  13. A376S in the connection subdomain of HIV-1 reverse transcriptase confers increased risk of virological failure to nevirapine therapy

    DEFF Research Database (Denmark)

    Paredes, Roger; Puertas, Maria Carmen; Bannister, Wendy

    2011-01-01

    Background. The clinical relevance of mutations in the connection subdomain and the ribonuclease (RNase) H domain of HIV-1 reverse transcriptase (RT) is uncertain. Methods. The risk of virological failure to nonnucleoside RT inhibitor (NNRTI)-based antiretroviral therapy (ART) was evaluated...

  14. HDAC inhibition induces HIV-1 protein and enables immune-based clearance following latency reversal

    DEFF Research Database (Denmark)

    Wu, Guoxin; Swanson, Michael; Talla, Aarthi

    2017-01-01

    , and the measurement of clearance of infected cells, is essential to assessing therapeutic efficacy. Here, we apply enhanced methodology extending the sensitivity limits for the rapid detection of subfemtomolar HIV gag p24 capsid protein in CD4+ T cells from ART-suppressed HIV+ individuals, and we show viral protein...... bispecific antibody. These findings extend beyond classical nucleic acid endpoints, which are confounded by the predominance of mutated, defective proviruses and, of paramount importance, enable assessment of cells making HIV protein that can now be targeted by immunological approaches.......Promising therapeutic approaches for eradicating HIV include transcriptional activation of provirus from latently infected cells using latency-reversing agents (LRAs) and immune-mediated clearance to purge reservoirs. Accurate detection of cells capable of producing viral antigens and virions...

  15. Design of Annulated Pyrazoles As Inhibitors of HIV-1 Reverse Transcriptase

    Energy Technology Data Exchange (ETDEWEB)

    Sweeney, Z.K.; Harris, S.F.; Arora, N.; Javanbakht, H.; Li, Y.; Fretland, J.; Davidson, J.P.; Billedeau, J.R.; Gleason, S.; Hirschfeld, D.; Kennedy-Smith, J.J.; Mirzadegan, T.; Roetz, R.; Smith, M.; Sperry, S.; Suh, J.M.; Wu, J.; Tsing, S.; Villasenor, A.G.; Paul, A.; Su, G.

    2009-05-26

    Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are recommended components of preferred combination antiretroviral therapies used for the treatment of HIV. These regimens are extremely effective in suppressing virus replication. Structure-based optimization of diaryl ether inhibitors led to the discovery of a new series of pyrazolo[3,4-c]pyridazine NNRTIs that bind the reverse transcriptase enzyme of human immunodeficiency virus-1 (HIV-RT) in an expanded volume relative to most other inhibitors in this class. The binding mode maintains the {beta}13 and {beta}14 strands bearing Pro236 in a position similar to that in the unliganded reverse transcriptase structure, and the distribution of interactions creates the opportunity for substantial resilience to single point mutations. Several pyrazolopyridazine NNRTIs were found to be highly effective against wild-type and NNRTI-resistant viral strains in cell culture.

  16. Comparative biochemical analysis of recombinant reverse transcriptase enzymes of HIV-1 subtype B and subtype C

    Directory of Open Access Journals (Sweden)

    Moisi Daniella

    2010-10-01

    Full Text Available Abstract Background HIV-1 subtype C infections account for over half of global HIV infections, yet the vast focus of HIV-1 research has been on subtype B viruses which represent less than 12% of the global pandemic. Since HIV-1 reverse transcriptase (RT is a major target of antiviral therapy, and since differential drug resistance pathways have been observed among different HIV subtypes, it is important to study and compare the enzymatic activities of HIV-1 RT derived from each of subtypes B and C as well as to determine the susceptibilities of these enzymes to various RT inhibitors in biochemical assays. Methods Recombinant subtype B and C HIV-1 RTs in heterodimeric form were purified from Escherichia coli and enzyme activities were compared in cell-free assays. The efficiency of (- ssDNA synthesis was measured using gel-based assays with HIV-1 PBS RNA template and tRNA3Lys as primer. Processivity was assayed under single-cycle conditions using both homopolymeric and heteropolymeric RNA templates. Intrinsic RNase H activity was compared using 5'-end labeled RNA template annealed to 3'-end recessed DNA primer in a time course study in the presence and absence of a heparin trap. A mis-incorporation assay was used to assess the fidelity of the two RT enzymes. Drug susceptibility assays were performed both in cell-free assays using recombinant enzymes and in cell culture phenotyping assays. Results The comparative biochemical analyses of recombinant subtype B and subtype C HIV-1 reverse transcriptase indicate that the two enzymes are very similar biochemically in efficiency of tRNA-primed (- ssDNA synthesis, processivity, fidelity and RNase H activity, and that both enzymes show similar susceptibilities to commonly used NRTIs and NNRTIs. Cell culture phenotyping assays confirmed these results. Conclusions Overall enzyme activity and drug susceptibility of HIV-1 subtype C RT are comparable to those of subtype B RT. The use of RT inhibitors (RTIs

  17. Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems.

    Directory of Open Access Journals (Sweden)

    Julia K Bialek

    Full Text Available CRISPR/Cas9 technology is currently considered the most advanced tool for targeted genome engineering. Its sequence-dependent specificity has been explored for locus-directed transcriptional modulation. Such modulation, in particular transcriptional activation, has been proposed as key approach to overcome silencing of dormant HIV provirus in latently infected cellular reservoirs. Currently available agents for provirus activation, so-called latency reversing agents (LRAs, act indirectly through cellular pathways to induce viral transcription. However, their clinical performance remains suboptimal, possibly because reservoirs have diverse cellular identities and/or proviral DNA is intractable to the induced pathways. We have explored two CRISPR/Cas9-derived activator systems as targeted approaches to induce dormant HIV-1 proviral DNA. These systems recruit multiple transcriptional activation domains to the HIV 5' long terminal repeat (LTR, for which we have identified an optimal target region within the LTR U3 sequence. Using this target region, we demonstrate transcriptional activation of proviral genomes via the synergistic activation mediator complex in various in culture model systems for HIV latency. Observed levels of induction are comparable or indeed higher than treatment with established LRAs. Importantly, activation is complete, leading to production of infective viral particles. Our data demonstrate that CRISPR/Cas9-derived technologies can be applied to counteract HIV latency and may therefore represent promising novel approaches in the quest for HIV elimination.

  18. HIV nonnucleoside reverse transcriptase inhibitors and trimethoprim-sulfamethoxazole inhibit plasmodium liver stages.

    Science.gov (United States)

    Hobbs, Charlotte V; Voza, Tatiana; De La Vega, Patricia; Vanvliet, Jillian; Conteh, Solomon; Penzak, Scott R; Fay, Michael P; Anders, Nicole; Ilmet, Tiina; Li, Yonghua; Borkowsky, William; Krzych, Urszula; Duffy, Patrick E; Sinnis, Photini

    2012-12-01

    Although nonnucleoside reverse transcriptase inhibitors (NNRTIs) are usually part of first-line treatment regimens for human immunodeficiency virus (HIV), their activity on Plasmodium liver stages remains unexplored. Additionally, trimethoprim-sulfamethoxazole (TMP-SMX), used for opportunistic infection prophylaxis in HIV-exposed infants and HIV-infected patients, reduces clinical episodes of malaria; however, TMP-SMX effect on Plasmodium liver stages requires further study. We characterized NNRTI and TMP-SMX effects on Plasmodium liver stages in vivo using Plasmodium yoelii. On the basis of these results, we conducted in vitro studies assessing TMP-SMX effects on the rodent parasites P. yoelii and Plasmodium berghei and on the human malaria parasite Plasmodium falciparum. Our data showed NNRTI treatment modestly reduced P. yoelii liver stage parasite burden and minimally extended prepatent period. TMP-SMX administration significantly reduced liver stage parasite burden, preventing development of patent parasitemia in vivo. TMP-SMX inhibited development of rodent and P. falciparum liver stage parasites in vitro. NNRTIs modestly affect liver stage Plasmodium parasites, whereas TMP-SMX prevents patent parasitemia. Because drugs that inhibit liver stages target parasites when they are present in lower numbers, these results may have implications for eradication efforts. Understanding HIV drug effects on Plasmodium liver stages will aid in optimizing treatment regimens for HIV-exposed and HIV-infected infected patients in malaria-endemic areas.

  19. Structure and function of HIV-1 reverse transcriptase: molecular mechanisms of polymerization and inhibition

    Science.gov (United States)

    Sarafianos, Stefan G.; Marchand, Bruno; Das, Kalyan; Himmel, Daniel; Parniak, Michael A.; Hughes, Stephen H.; Arnold, Eddy

    2009-01-01

    The rapid replication of HIV-1 and the errors made during viral replication, cause the virus to evolve rapidly in patients, making the problems of vaccine development and drug therapy particularly challenging. In the absence of an effective vaccine, drugs are the only useful treatment. Anti-HIV drugs work; so far drug therapy has saved more than three million years of life. Unfortunately, HIV-1 develops resistance to all of the available drugs. Although a number of useful anti-HIV drugs have been approved for use in patients, the problems associated with drug toxicity and the development of resistance means that the search for new drugs is an ongoing process. The three viral enzymes, reverse transcriptase (RT), integrase (IN), and protease (PR) are all good drug targets. Two distinct types of RT inhibitors, both of which block the polymerase activity of RT, have been approved to treat HIV-1 infections, nucleoside analogs (NRTIs) and nonnucleosides (NNRTIs), and there are promising leads for compounds that either block the RNase H activity or block the polymerase in other ways. A better understanding of the structure and function(s) of RT and of the mechanism(s) of inhibition can be used to generate better drugs; in particular drugs that are effective against the current drug-resistant strains of HIV-1. PMID:19022262

  20. Reverse electrodialysis : evaluation of suitable electrode systems

    NARCIS (Netherlands)

    Veerman, J.; Saakes, M.; Metz, S. J.; Harmsen, G. J.

    Reverse electrodialysis (RED) is a method for directly extracting electrical energy from salinity gradients, especially from sea and river water. For the commercial implementation of RED, the electrode system is a key component. In this paper, novel electrode systems for RED were compared with

  1. An HIV-1 Replication Pathway Utilizing Reverse Transcription Products That Fail To Integrate

    Science.gov (United States)

    Trinité, Benjamin; Ohlson, Eric C.; Voznesensky, Igor; Rana, Shashank P.; Chan, Chi N.; Mahajan, Saurabh; Alster, Jason; Burke, Sean A.; Wodarz, Dominik

    2013-01-01

    Integration is a central event in the replication of retroviruses, yet ≥90% of HIV-1 reverse transcripts fail to integrate, resulting in accumulation of unintegrated viral DNA in cells. However, understanding what role, if any, unintegrated viral DNA plays in the natural history of HIV-1 has remained elusive. Unintegrated HIV-1 DNA is reported to possess a limited capacity for gene expression restricted to early gene products and is considered a replicative dead end. Although the majority of peripheral blood CD4+ T cells are refractory to infection, nonactivated CD4 T cells present in lymphoid and mucosal tissues are major targets for infection. Treatment with cytokine interleukin-2 (IL-2), IL-4, IL-7, or IL-15 renders CD4+ T cells permissive to HIV-1 infection in the absence of cell activation and proliferation and provides a useful model for infection of resting CD4+ T cells. We found that infection of cytokine-treated resting CD4+ T cells in the presence of raltegravir or with integrase active-site mutant HIV-1 yielded de novo virus production following subsequent T cell activation. Infection with integration-competent HIV-1 naturally generated a population of cells generating virus from unintegrated DNA. Latent infection persisted for several weeks and could be activated to virus production by a combination of a histone deacetylase inhibitor and a protein kinase C activator or by T cell activation. HIV-1 Vpr was essential for unintegrated HIV-1 gene expression and de novo virus production in this system. Bypassing integration by this mechanism may allow the preservation of genetic information that otherwise would be lost. PMID:24049167

  2. Data on synthesis of methylene bisphosphonates and screening of their inhibitory activity towards HIV reverse transcriptase

    Directory of Open Access Journals (Sweden)

    D.V. Yanvarev

    2016-09-01

    Full Text Available Inorganic pyrophosphate (PPi mimetics designed on a basis of methylenediphosphonic acid backbone are promising inhibitors of two key HIV replication enzymes, IN [1] and RT [2]. Herein, we present chemical synthesis of eleven methylenebisphosphonates (BPs with their NMR and HRMS analysis synthesized via five different ways. Also, we present data on inhibition of HIV RT catalyzed phosphorolysis and polymerization by synthesized BPs using two methods based on denaturing urea PAGE. Tests were also performed for thymidine analogue mutations reverse transcriptase (TAM RT, which was expressed and purified for that. Structure–activity relationships and inhibitory activity data of synthesized BPs are presented in “Methylene bisphosphonates as the inhibitors of HIV RT phosphorolytic activity” [2].

  3. The Role of Nucleotide Excision by Reverse Transcriptase in HIV Drug Resistance

    Directory of Open Access Journals (Sweden)

    Walter A. Scott

    2010-01-01

    Full Text Available Nucleoside reverse transcriptase (RT inhibitors of HIV block viral replication through the ability of HIV RT to incorporate chain-terminating nucleotide analogs during viral DNA synthesis. Once incorporated, the chain-terminating residue must be removed before DNA synthesis can continue. Removal can be accomplished by the excision activity of HIV RT, which catalyzes the transfer of the 3'-terminal residue on the blocked DNA chain to an acceptor substrate, probably ATP in most infected cells. Mutations of RT that enhance excision activity are the most common cause of resistance to 3'-azido-3'-deoxythymidine (AZT and exhibit low-level cross-resistance to most other nucleoside RT inhibitors. The resistance to AZT is suppressed by a number of additional mutations in RT, most of which were identified because they conferred resistance to other RT inhibitors. Here we review current understanding of the biochemical mechanisms responsible for increased or decreased excision activity due to these mutations.

  4. Structural Aspects of Drug Resistance and Inhibition of HIV-1 Reverse Transcriptase

    Directory of Open Access Journals (Sweden)

    Stefan G. Sarafianos

    2010-02-01

    Full Text Available HIV-1 Reverse Transcriptase (HIV-1 RT has been the target of numerous approved anti-AIDS drugs that are key components of Highly Active Anti-Retroviral Therapies (HAART. It remains the target of extensive structural studies that continue unabated for almost twenty years. The crystal structures of wild-type or drug-resistant mutant HIV RTs in the unliganded form or in complex with substrates and/or drugs have offered valuable glimpses into the enzyme’s folding and its interactions with DNA and dNTP substrates, as well as with nucleos(tide reverse transcriptase inhibitor (NRTI and non-nucleoside reverse transcriptase inhibitor (NNRTIs drugs. These studies have been used to interpret a large body of biochemical results and have paved the way for innovative biochemical experiments designed to elucidate the mechanisms of catalysis and drug inhibition of polymerase and RNase H functions of RT. In turn, the combined use of structural biology and biochemical approaches has led to the discovery of novel mechanisms of drug resistance and has contributed to the design of new drugs with improved potency and ability to suppress multi-drug resistant strains.

  5. Evaluating Reverse Supply Chain Efficiency: Manufacturer's Perspective

    DEFF Research Database (Denmark)

    Kumar, M.; Tiwari, M. K.; Wong, K. Y.

    2014-01-01

    The paper aims to illustrate the use of fuzzy data envelopment analysis (DEA) in analyzing reverse supply chain (RSC) performance from the manufacturer's perspective. By using an alternative alpha-cut approach, the fuzzy DEA model was converted into a crisp linear programming problem, thereby...... altering the problem to an interval programming one. The model is able to obtain precise and robust efficiency values. An investigation was also made between the obtained efficiency scores and certain relevant background information of the companies. The study revealed that while ISO 14001 certification...

  6. Naringin Reverses Hepatocyte Apoptosis and Oxidative Stress Associated with HIV-1 Nucleotide Reverse Transcriptase Inhibitors-Induced Metabolic Complications

    Directory of Open Access Journals (Sweden)

    Oluwafeyisetan O. Adebiyi

    2015-12-01

    Full Text Available Nucleoside Reverse Transcriptase Inhibitors (NRTIs have not only improved therapeutic outcomes in the treatment of HIV infection but have also led to an increase in associated metabolic complications of NRTIs. Naringin’s effects in mitigating NRTI-induced complications were investigated in this study. Wistar rats, randomly allotted into seven groups (n = 7 were orally treated daily for 56 days with 100 mg/kg zidovudine (AZT (groups I, II III, 50 mg/kg stavudine (d4T (groups IV, V, VI and 3 mL/kg of distilled water (group VII. Additionally, rats in groups II and V were similarly treated with 50 mg/kg naringin, while groups III and VI were treated with 45 mg/kg vitamin E. AZT or d4T treatment significantly reduced body weight and plasma high density lipoprotein concentrations but increased liver weights, plasma triglycerides and total cholesterol compared to controls, respectively. Furthermore, AZT or d4T treatment significantly increased oxidative stress, adiposity index and expression of Bax protein, but reduced Bcl-2 protein expression compared to controls, respectively. However, either naringin or vitamin E significantly mitigated AZT- or d4T-induced weight loss, dyslipidemia, oxidative stress and hepatocyte apoptosis compared to AZT- or d4T-only treated rats. Our results suggest that naringin reverses metabolic complications associated with NRTIs by ameliorating oxidative stress and apoptosis. This implies that naringin supplements could mitigate lipodystrophy and dyslipidemia associated with NRTI therapy.

  7. Chronic toxicological evaluation and reversibility studies of Moringa ...

    African Journals Online (AJOL)

    Chronic toxicological evaluation and reversibility studies of Moringa oleifera ethanolic seed extract in Wistar rats. ... of the animals were sacrificed and blood samples collected for determination of biochemical and haematological parameters; antioxidants and malondialdehyde (MDA); sperm count, motility and morphology.

  8. Evaluation of the Determine™ fourth generation HIV rapid assay.

    Science.gov (United States)

    Brauer, Marieke; De Villiers, Johanna C; Mayaphi, Simnikiwe H

    2013-04-01

    Assays that detect p24 antigen reduce the diagnostic window period of HIV testing. Most point-of-care HIV assays have poor sensitivity to diagnose acute HIV infection as they only detect antibodies against HIV-1 and HIV-2 (HIV-1/2). This was a cross-sectional laboratory-based study that evaluated the performance of the Determine™ HIV-1/2 Ag/Ab Combo fourth generation rapid strip - currently the only rapid assay that detects both HIV-1/2 antibodies and p24 antigen. A total of 79 serum specimens (29 positive for HIV antibodies only, 14 positive for HIV antibodies and p24 antigen, 20 HIV-negative, and 16 positive for p24 antigen only) were used for the evaluation. Results were compared with those from validated fourth generation HIV ELISAs. The Determine™ Combo rapid strips had a sensitivity of 90.7% and a specificity of 100% for the detection of HIV-1/2 antibodies. Its sensitivity for the detection of p24 antigen was only 10% (3 out of 30 p24 antigen positive specimens). This implies that most acute HIV infections will be missed with this assay. The need for a point-of-care assay which can detect acute HIV infection reliably still remains, particularly for use in a high prevalence setting such as South Africa. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Initiation of ART during early acute HIV infection preserves mucosal Th17 function and reverses HIV-related immune activation.

    Directory of Open Access Journals (Sweden)

    Alexandra Schuetz

    2014-12-01

    Full Text Available Mucosal Th17 cells play an important role in maintaining gut epithelium integrity and thus prevent microbial translocation. Chronic HIV infection is characterized by mucosal Th17 cell depletion, microbial translocation and subsequent immune-activation, which remain elevated despite antiretroviral therapy (ART correlating with increased mortality. However, when Th17 depletion occurs following HIV infection is unknown. We analyzed mucosal Th17 cells in 42 acute HIV infection (AHI subjects (Fiebig (F stage I-V with a median duration of infection of 16 days and the short-term impact of early initiation of ART. Th17 cells were defined as IL-17+ CD4+ T cells and their function was assessed by the co-expression of IL-22, IL-2 and IFNγ. While intact during FI/II, depletion of mucosal Th17 cell numbers and function was observed during FIII correlating with local and systemic markers of immune-activation. ART initiated at FI/II prevented loss of Th17 cell numbers and function, while initiation at FIII restored Th17 cell numbers but not their polyfunctionality. Furthermore, early initiation of ART in FI/II fully reversed the initially observed mucosal and systemic immune-activation. In contrast, patients treated later during AHI maintained elevated mucosal and systemic CD8+ T-cell activation post initiation of ART. These data support a loss of Th17 cells at early stages of acute HIV infection, and highlight that studies of ART initiation during early AHI should be further explored to assess the underlying mechanism of mucosal Th17 function preservation.

  10. Evaluation of HIV therapeutic agents on immunological, lipid and ...

    African Journals Online (AJOL)

    HIV-1 infected patients initiating antiretroviral therapy (ART) in Ghana are placed on one of the two most commonly used non-nucleoside reverse transcriptase inhibitors (NNRTIs), nevirapine (NVP) and efavirenz (EFV), in combination with a nucleoside reverse transcriptase inhibitor backbone of either combivir (CBV) or ...

  11. Structural and Preclinical Studies of Computationally Designed Non-Nucleoside Reverse Transcriptase Inhibitors for Treating HIV infection

    Energy Technology Data Exchange (ETDEWEB)

    Kudalkar, Shalley N.; Beloor, Jagadish; Chan, Albert H.; Lee, Won-Gil; Jorgensen, William L.; Kumar, Priti; Anderson, Karen S.

    2017-02-06

    The clinical benefits of HIV-1 non-nucleoside reverse transcriptase (RT) inhibitors (NNRTIs) are hindered by their unsatisfactory pharmacokinetic (PK) properties along with the rapid development of drug-resistant variants. However, the clinical efficacy of these inhibitors can be improved by developing compounds with enhanced pharmacological profiles and heightened antiviral activity. We used computational and structure-guided design to develop two next-generation NNRTI drug candidates, compounds I and II, which are members of a class of catechol diethers. We evaluated the preclinical potential of these compounds in BALB/c mice because of their high solubility (510 µg/ml for compound I and 82.9 µg/ml for compound II), low cytotoxicity, and enhanced antiviral activity against wild-type (WT) HIV-1 RT and resistant variants. Additionally, crystal structures of compounds I and II with WT RT suggested an optimal binding to the NNRTI binding pocket favoring the high anti-viral potency. A single intraperitoneal dose of compounds I and II exhibited a prolonged serum residence time of 48 hours and concentration maximum (Cmax) of 4000- to 15,000-fold higher than their therapeutic/effective concentrations. These Cmax values were 4- to 15-fold lower than their cytotoxic concentrations observed in MT-2 cells. Compound II showed an enhanced area under the curve (0–last) and decreased plasma clearance over compound I and efavirenz, the standard of care NNRTI. Hence, the overall (PK) profile of compound II was excellent compared with that of compound I and efavirenz. Furthermore, both compounds were very well tolerated in BALB/c mice without any detectable acute toxicity. Taken together, these data suggest that compounds I and II possess improved anti-HIV-1 potency, remarkable in vivo safety, and prolonged in vivo circulation time, suggesting strong potential for further development as new NNRTIs for the potential treatment of HIV infection.

  12. The Lysine 65 Residue in HIV-1 Reverse Transcriptase Function and in Nucleoside Analog Drug Resistance

    Directory of Open Access Journals (Sweden)

    Scott J. Garforth

    2014-10-01

    Full Text Available Mutations in HIV-1 reverse transcriptase (RT that confer nucleoside analog RT inhibitor resistance have highlighted the functional importance of several active site residues (M184, Q151 and K65 in RT catalytic function. Of these, K65 residue is notable due to its pivotal position in the dNTP-binding pocket, its involvement in nucleoside analog resistance and polymerase fidelity. This review focuses on K65 residue and summarizes a substantial body of biochemical and structural studies of its role in RT function and the functional consequences of the K65R mutation.

  13. Novel 2-Chloro-8-arylthiomethyldipyridodiazepinone Derivatives with Activity against HIV-1 Reverse Transcriptase

    Directory of Open Access Journals (Sweden)

    Supanna Techasakul

    2007-02-01

    Full Text Available Based on the molecular modeling analysis against Y181CHIV-1 RT, dipyridodiazepinone derivatives containing an unsubstituted lactamnitrogen and 2-chloro-8-arylthiomethyl were synthesized via an efficientroute. Some of them were evaluated for their antiviral activity against HIV-1RT subtype E and were found to exhibit virustatic activity comparable to some clinically usedtherapeutic agents.

  14. [Evaluation of Abbott Fourth Generation HIV Antigen and Antibody Assays.].

    Science.gov (United States)

    Kang, Hee Jung; Yoo, Kyeong Ha; Kim, Han Sung; Cho, Hyoun Chan

    2006-02-01

    In order to reduce the diagnostic window period between the time of human immunodeficiency virus (HIV) infection and serological diagnosis, new fourth generation screening assays which detect HIV p24 antigen and specific antibody simultaneously have been developed. In this study, we evaluated the performance of a new fourth generation assay. We compared a new fourth generation assay, Architect HIV Ag/Ab combo, with another fourth generation assay AxSYM HIV Ag/Ab combo and a third generation assay, AxSYM HIV 1/2 gO for their performance. The assays were evaluated using 3 HIV seroconversion panels, 305 sera of healthy subjects and 100 sera of patients with HBsAg or anti-HCV antibodies. Within-run and total coefficient variations of the three screening assays were analyzed for the evaluation of precision. Architect HIV Ag/Ab combo shortened the window period by 8.7+/-2.1 days relative to AxSYM HIV 1/2 gO and 2.0+/-2.0 days relative to AxSYM HIV Ag/Ab combo in seroconversion panels. Architect HIV Ag/Ab combo presented the best performance in precision among the three reagents; total CV for positive control was 3.6%, 9.6% and 4.6% for Architect HIV Ag/Ab combo, AxSYM HIV Ag/Ab combo and AxSYM HIV 1/2 gO, respectively. Specificities of three assays were not different in this study. HIV Ag/Ab combined assays reduced the diagnostic window as compared to the third generation screening assays, enabling an earlier diagnosis of HIV infection. A new fourth generation assay, Architect HIV Ag/Ab combo presents a better performance than AxSYM HIV Ag/Ab combo, showing improved seroconversion sensitivity and precision.

  15. Prediction of mutational tolerance in HIV-1 protease and reverse transcriptase using flexible backbone protein design.

    Directory of Open Access Journals (Sweden)

    Elisabeth Humphris-Narayanan

    Full Text Available Predicting which mutations proteins tolerate while maintaining their structure and function has important applications for modeling fundamental properties of proteins and their evolution; it also drives progress in protein design. Here we develop a computational model to predict the tolerated sequence space of HIV-1 protease reachable by single mutations. We assess the model by comparison to the observed variability in more than 50,000 HIV-1 protease sequences, one of the most comprehensive datasets on tolerated sequence space. We then extend the model to a second protein, reverse transcriptase. The model integrates multiple structural and functional constraints acting on a protein and uses ensembles of protein conformations. We find the model correctly captures a considerable fraction of protease and reverse-transcriptase mutational tolerance and shows comparable accuracy using either experimentally determined or computationally generated structural ensembles. Predictions of tolerated sequence space afforded by the model provide insights into stability-function tradeoffs in the emergence of resistance mutations and into strengths and limitations of the computational model.

  16. Crystal Engineering of HIV-1 Reverse Transcriptase for structure-Based Drug Design

    Energy Technology Data Exchange (ETDEWEB)

    Bauman,J.; Das, K.; Ho, W.; Baweja, M.; Himmel, D.; Clark, A.; Oren, D.; Shatkin, A.; Arnold, E.

    2008-01-01

    HIV-1 reverse transcriptase (RT) is a primary target for anti-AIDS drugs. Structures of HIV-1 RT, usually determined at {approx}2.5-3.0 Angstroms resolution, are important for understanding enzyme function and mechanisms of drug resistance in addition to being helpful in the design of RT inhibitors. Despite hundreds of attempts, it was not possible to obtain the structure of a complex of HIV-1 RT with TMC278, a nonnucleoside RT inhibitor (NNRTI) in advanced clinical trials. A systematic and iterative protein crystal engineering approach was developed to optimize RT for obtaining crystals in complexes with TMC278 and other NNRTIs that diffract X-rays to 1.8 Angstroms resolution. Another form of engineered RT was optimized to produce a high-resolution apo-RT crystal form, reported here at 1.85 Angstroms resolution, with a distinct RT conformation. Engineered RTs were mutagenized using a new, flexible and cost effective method called methylated overlap-extension ligation independent cloning. Our analysis suggests that reducing the solvent content, increasing lattice contacts, and stabilizing the internal low-energy conformations of RT are critical for the growth of crystals that diffract to high resolution. The new RTs enable rapid crystallization and yield high-resolution structures that are useful in designing/developing new anti-AIDS drugs.

  17. Global Conformational Dynamics of HIV-1 Reverse Transcriptase Bound to Non-Nucleoside Inhibitors

    Directory of Open Access Journals (Sweden)

    Peter V. Coveney

    2012-07-01

    Full Text Available HIV-1 Reverse Transcriptase (RT is a multifunctional enzyme responsible for the transcription of the RNA genome of the HIV virus into DNA suitable for incorporation within the DNA of human host cells. Its crucial role in the viral life cycle has made it one of the major targets for antiretroviral drug therapy. The Non-Nucleoside RT Inhibitor (NNRTI class of drugs binds allosterically to the enzyme, affecting many aspects of its activity. We use both coarse grained network models and atomistic molecular dynamics to explore the changes in protein dynamics induced by NNRTI binding. We identify changes in the flexibility and conformation of residue Glu396 in the RNaseH primer grip which could provide an explanation for the acceleration in RNaseH cleavage rate observed experimentally in NNRTI bound HIV-1 RT. We further suggest a plausible path for conformational and dynamic changes to be communicated from the vicinity of the NNRTI binding pocket to the RNaseH at the other end of the enzyme.

  18. Evaluation of reversible contraceptive activities of methanol extract ...

    African Journals Online (AJOL)

    The present study was undertaken to evaluate the contraceptive activities of methanol stem bark extract of Annoa squamosa L. (Annonaceae) with their respective reversibility in male rats. Proven fertile male rats were gavaged 100% methanol extract of A. squamosa stem bark at the dose level of 50, 100 and 200 mg/rat/day ...

  19. Compounds acting against HIV: Imidazo[1,2-a]pyridines as non-nucleoside reverse transcriptase inhibitors (NNRTIs)

    CSIR Research Space (South Africa)

    Bode, M

    2010-09-01

    Full Text Available A compound possessing anti-HIV activity similar to that of FDA-approved nevirapine was developed. It acts as a non-nucleoside reverse transcriptase inhibitor, the compound shows good cell permeability, and a quantitative structure activity...

  20. Crystal structure of HIV-1 reverse transcriptase in complex with a polypurine tract RNA:DNA

    Science.gov (United States)

    Sarafianos, Stefan G.; Das, Kalyan; Tantillo, Chris; Clark, Arthur D.; Ding, Jianping; Whitcomb, Jeannette M.; Boyer, Paul L.; Hughes, Stephen H.; Arnold, Edward

    2001-01-01

    We have determined the 3.0 Å resolution structure of wild-type HIV-1 reverse transcriptase in complex with an RNA:DNA oligonucleotide whose sequence includes a purine-rich segment from the HIV-1 genome called the polypurine tract (PPT). The PPT is resistant to ribonuclease H (RNase H) cleavage and is used as a primer for second DNA strand synthesis. The ‘RNase H primer grip’, consisting of amino acids that interact with the DNA primer strand, may contribute to RNase H catalysis and cleavage specificity. Cleavage specificity is also controlled by the width of the minor groove and the trajectory of the RNA:DNA, both of which are sequence dependent. An unusual ‘unzipping’ of 7 bp occurs in the adenine stretch of the PPT: an unpaired base on the template strand takes the base pairing out of register and then, following two offset base pairs, an unpaired base on the primer strand re-establishes the normal register. The structural aberration extends to the RNase H active site and may play a role in the resistance of PPT to RNase H cleavage. PMID:11250910

  1. Structure of the HIV-1 reverse transcriptase Q151M mutant: insights into the inhibitor resistance of HIV-1 reverse transcriptase and the structure of the nucleotide-binding pocket of Hepatitis B virus polymerase

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Akiyoshi; Tamura, Noriko; Yasutake, Yoshiaki, E-mail: y-yasutake@aist.go.jp [National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo, Hokkaido 062-8517 (Japan)

    2015-10-23

    The structure of the HIV-1 reverse transcriptase Q151M mutant was determined at a resolution of 2.6 Å in space group P321. Hepatitis B virus polymerase (HBV Pol) is an important target for anti-HBV drug development; however, its low solubility and stability in vitro has hindered detailed structural studies. Certain nucleotide reverse transcriptase (RT) inhibitors (NRTIs) such as tenofovir and lamivudine can inhibit both HBV Pol and Human immunodeficiency virus 1 (HIV-1) RT, leading to speculation on structural and mechanistic analogies between the deoxynucleotide triphosphate (dNTP)-binding sites of these enzymes. The Q151M mutation in HIV-1 RT, located at the dNTP-binding site, confers resistance to various NRTIs, while maintaining sensitivity to tenofovir and lamivudine. The residue corresponding to Gln151 is strictly conserved as a methionine in HBV Pol. Therefore, the structure of the dNTP-binding pocket of the HIV-1 RT Q151M mutant may reflect that of HBV Pol. Here, the crystal structure of HIV-1 RT Q151M, determined at 2.6 Å resolution, in a new crystal form with space group P321 is presented. Although the structure of HIV-1 RT Q151M superimposes well onto that of HIV-1 RT in a closed conformation, a slight movement of the β-strands (β2–β3) that partially create the dNTP-binding pocket was observed. This movement might be caused by the introduction of the bulky thioether group of Met151. The structure also highlighted the possibility that the hydrogen-bonding network among amino acids and NRTIs is rearranged by the Q151M mutation, leading to a difference in the affinity of NRTIs for HIV-1 RT and HBV Pol.

  2. Conservation of functional domains and limited heterogeneity of HIV-1 reverse transcriptase gene following vertical transmission

    Directory of Open Access Journals (Sweden)

    Ahmad Nafees

    2005-05-01

    Full Text Available Abstract Background The reverse transcriptase (RT enzyme of human immunodeficiency virus type 1 (HIV-1 plays a crucial role in the life cycle of the virus by converting the single stranded RNA genome into double stranded DNA that integrates into the host chromosome. In addition, RT is also responsible for the generation of mutations throughout the viral genome, including in its own sequences and is thus responsible for the generation of quasi-species in HIV-1-infected individuals. We therefore characterized the molecular properties of RT, including the conservation of functional motifs, degree of genetic diversity, and evolutionary dynamics from five mother-infant pairs following vertical transmission. Results The RT open reading frame was maintained with a frequency of 87.2% in five mother-infant pairs' sequences following vertical transmission. There was a low degree of viral heterogeneity and estimates of genetic diversity in mother-infant pairs' sequences. Both mothers and infants RT sequences were under positive selection pressure, as determined by the ratios of non-synonymous to synonymous substitutions. Phylogenetic analysis of 132 mother-infant RT sequences revealed distinct clusters for each mother-infant pair, suggesting that the epidemiologically linked mother-infant pairs were evolutionarily closer to each other as compared with epidemiologically unlinked mother-infant pairs. The functional domains of RT which are responsible for reverse transcription, DNA polymerization and RNase H activity were mostly conserved in the RT sequences analyzed in this study. Specifically, the active sites and domains required for primer binding, template binding, primer and template positioning and nucleotide recruitment were conserved in all mother-infant pairs' sequences. Conclusion The maintenance of an intact RT open reading frame, conservation of functional domains for RT activity, preservation of several amino acid motifs in epidemiologically

  3. Performance of 3 Rapid Tests for Discrimination Between HIV-1 and HIV-2 in Guinea-Bissau, West Africa

    DEFF Research Database (Denmark)

    Hønge, Bo Langhoff; Bjarnason Obinah, Magnús Pétur; Jespersen, Sanne

    2014-01-01

    As HIV-2 is intrinsically resistant to nonnucleoside reverse transcriptase inhibitors, it is mandatory to discriminate between HIV types before initiating antiretroviral treatment. Guinea-Bissau has the world's highest prevalence of HIV-2 and HIV-1/HIV-2 dually infected individuals. We evaluated ...

  4. Evaluation neuer Methoden zur HIV-Inzidenztestung

    OpenAIRE

    Han, Orjin

    2014-01-01

    Hintergrund: Die Kenntnis der HIV-Neuinfektionen und ihrem Anteil in einer Bevölkerung (HIV-Inzidenz) ist für die epidemiologische Überwachung von HIV (HIV-Surveillance) von zentraler Bedeutung. Dadurch kann die Epidemie in einer Population charakterisiert werden, indem die Trends im Infektionsgeschehen aufgezeigt und wichtige Risikogruppen identifiziert werden, um die entsprechenden Präventionsmaßnahmen vorzunehmen. Zur Unterscheidung von kürzlich erworbenen und länger bestehenden HIV-Infek...

  5. hiv

    African Journals Online (AJOL)

    2016-03-31

    Mar 31, 2016 ... Indexed By: African Journal Online (AJOL); Texila American University; Genamics; Scholarsteer; EIJASR; CAS-American Chemical. Society; and IRMS Informatics India (J-Gate). ABSTRACT. This study evaluated the effect of HIV infection on CD4 T-lymphocyte depletion in people living with HIV/AIDS.

  6. Integrating Community-Based Interventions to Reverse the Convergent TB/HIV Epidemics in Rural South Africa

    Science.gov (United States)

    Gilbert, Jennifer A.; Long, Elisa F.; Brooks, Ralph P.; Friedland, Gerald H.; Moll, Anthony P.; Townsend, Jeffrey P.; Galvani, Alison P.; Shenoi, Sheela V.

    2015-01-01

    The WHO recommends integrating interventions to address the devastating TB/HIV co-epidemics in South Africa, yet integration has been poorly implemented and TB/HIV control efforts need strengthening. Identifying infected individuals is particularly difficult in rural settings. We used mathematical modeling to predict the impact of community-based, integrated TB/HIV case finding and additional control strategies on South Africa’s TB/HIV epidemics. We developed a model incorporating TB and HIV transmission to evaluate the effectiveness of integrating TB and HIV interventions in rural South Africa over 10 years. We modeled the impact of a novel screening program that integrates case finding for TB and HIV in the community, comparing it to status quo and recommended TB/HIV control strategies, including GeneXpert, MDR-TB treatment decentralization, improved first-line TB treatment cure rate, isoniazid preventive therapy, and expanded ART. Combining recommended interventions averted 27% of expected TB cases (95% CI 18–40%) 18% HIV (95% CI 13–24%), 60% MDR-TB (95% CI 34–83%), 69% XDR-TB (95% CI 34–90%), and 16% TB/HIV deaths (95% CI 12–29). Supplementing these interventions with annual community-based TB/HIV case finding averted a further 17% of TB cases (44% total; 95% CI 31–56%), 5% HIV (23% total; 95% CI 17–29%), 8% MDR-TB (68% total; 95% CI 40–88%), 4% XDR-TB (73% total; 95% CI 38–91%), and 8% TB/HIV deaths (24% total; 95% CI 16–39%). In addition to increasing screening frequency, we found that improving TB symptom questionnaire sensitivity, second-line TB treatment delays, default before initiating TB treatment or ART, and second-line TB drug efficacy were significantly associated with even greater reductions in TB and HIV cases. TB/HIV epidemics in South Africa were most effectively curtailed by simultaneously implementing interventions that integrated community-based TB/HIV control strategies and targeted drug-resistant TB. Strengthening

  7. Evaluation of HIV Surveillance System in Rivers State, Nigeria ...

    African Journals Online (AJOL)

    Background: Rivers State has been reported to have the highest HIV prevalence of all the thirty-six states in Nigeria. HIV surveillance system generates information for timely and appropriate public health action. Evaluation of the surveillance system is vital in ensuring that the purpose of the surveillance system is being met.

  8. HIV reverse transcriptase gene mutations in anti-retroviral treatment naïve rural people living with HIV/AIDS

    Directory of Open Access Journals (Sweden)

    K Mohanakrishnan

    2015-01-01

    Full Text Available This study is designed to find out the mutational variations of reverse transcriptase (RT gene of HIV, after the traditional drug usage among anti-retroviral therapy naïve rural people living with HIV/AIDS. HIV Reactive patients, who were exposed for indigenous medicines such as Siddha, Ayurveda etc., for a minimum period of 6 months were taken for this study. Among 40 patients, two samples (5.55% demonstrated high-level mutational resistance variations for nucleoside RT inhibitor (NRTI and non-NRTI. The predominant polymorphisms detected were K122E (91.7%, V60I (91.7%, V35T (89%, Q207E (89%, D177E (89%, T200A (86.1%, S48T (83.33%, K173A (80.6%.

  9. Efficient in vitro inhibition of HIV-1 gag reverse transcription by peptide nucleic acid (PNA) at minimal ratios of PNA/RNA

    DEFF Research Database (Denmark)

    Koppelhus, Uffe; Zachar, Vladimir; Nielsen, P.E.

    1997-01-01

    ) was investigated. We found that a bis-PNA (parallel antisense 10mer linked to antiparallel antisense 10mer) was superior to both the parallel antisense 10mer and antiparallel antisense 10mer in inhibiting reverse transcription of the gene, thus indicating triplex formation at the target sequence. A complete arrest......We have tested the inhibitory potential of peptide nucleic acid (PNA) on in vitro reverse transcription of the HIV-1 gag gene. PNA was designed to target different regions of the HIV-1 gag gene and the effect on reverse transcription by HIV-1, MMLV and AMV reverse transcriptases (RTs...... of reverse transcription was obtained at approximately 6-fold molar excess of the bis-PNA with respect to the gag RNA. At this molar ratio we found no effect on in vitro translation of gag RNA. A 15mer duplex-forming PNA was also found to inhibit reverse transcription at very low molar ratios of PNA/ gag RNA...

  10. Primary T-lymphocytes rescue the replication of HIV-1 DIS RNA mutants in part by facilitating reverse transcription.

    Science.gov (United States)

    Jones, Kate L; Sonza, Secondo; Mak, Johnson

    2008-03-01

    The dimerization initiation site (DIS) stem-loop within the HIV-1 RNA genome is vital for the production of infectious virions in T-cell lines but not in primary cells. In comparison to peripheral blood mononuclear cells (PBMCs), which can support the replication of both wild type and HIV-1 DIS RNA mutants, we have found that DIS RNA mutants are up to 100 000-fold less infectious than wild-type HIV-1 in T-cell lines. We have also found that the cell-type-dependent replication of HIV-1 DIS RNA mutants is largely producer cell-dependent, with mutants displaying a greater defect in viral cDNA synthesis when viruses were not derived from PBMCs. While many examples exist of host-pathogen interplays that are mediated via proteins, analogous examples which rely on nucleic acid triggers are limited. Our data provide evidence to illustrate that primary T-lymphocytes rescue, in part, the replication of HIV-1 DIS RNA mutants through mediating the reverse transcription process in a cell-type-dependent manner. Our data also suggest the presence of a host cell factor that acts within the virus producer cells. In addition to providing an example of an RNA-mediated cell-type-dependent block to viral replication, our data also provides evidence which help to resolve the dilemma of how HIV-1 genomes with mismatched DIS sequences can recombine to generate chimeric viral RNA genomes.

  11. Evaluation of the HIV-1 reverse transcriptase inhibitory properties of ...

    African Journals Online (AJOL)

    Geoffrey M Rukunga, Mawuli W Kofi-Tsekpo, Masahiko Kurokawa, Seiji Kageyama, Geoffrey M Mungai, John M Muli, Festus M Tolo, Rukia M Kibaya, Charles N Muthaura, James N Kanyara, Peter M Tukei, Kimiyasu Shiraki. Abstract. Extracts from twenty two medicinal plants popularly used in preparing traditional remedies ...

  12. Oxidative stress induced by HIV-1 reverse transcriptase modulates the enzyme's performance in gene immunization.

    Science.gov (United States)

    Isaguliants, Maria; Smirnova, Olga; Ivanov, Alexander V; Kilpelainen, Athina; Kuzmenko, Yulia; Petkov, Stefan; Latanova, Anastasia; Krotova, Olga; Engström, Gunnel; Karpov, Vadim; Kochetkov, Sergey; Wahren, Britta; Starodubova, Elizaveta

    2013-10-01

    HIV-1 infection induces chronic oxidative stress. The resultant neurotoxicity has been associated with Tat protein. Here, we for the first time describe the induction of oxidative stress by another HIV-1 protein, reverse transcriptase (RT). Expression of HIV-1 RT in human embryonic kidney cells generated potent production of the reactive oxygen species (ROS), detected by the fluorescence-based probes. Quantitative RT-PCR demonstrated that expression of RT in HEK293 cells induced a 10- to 15-fold increased transcription of the phase II detoxifying enzymes human quinone oxidoreductase (Nqo1) and heme oxygenase 1 (HO-1), indicating the induction of oxidative stress response. The capacity to induce oxidative stress and stress response appeared to be an intrinsic property of a vast variety of RTs: enzymatically active and inactivated, bearing mutations of drug resistance, following different routes of processing and presentation, expressed from viral or synthetic expression-optimized genes. The total ROS production induced by RT genes of the viral origin was found to be lower than that induced by the synthetic/expression-optimized or chimeric RT genes. However, the viral RT genes induced higher levels of ROS production and higher levels of HO-1 mRNA than the synthetic genes per unit of protein in the expressing cell. The capacity of RT genes to induce the oxidative stress and stress response was then correlated with their immunogenic performance. For this, RT genes were administered into BALB/c mice by intradermal injections followed by electroporation. Splenocytes of immunized mice were stimulated with the RT-derived and control antigens and antigen-specific proliferation was assessed by IFN-γ/IL-2 Fluorospot. RT variants generating high total ROS levels induced significantly stronger IFN-γ responses than the variants inducing lower total ROS, while high levels of ROS normalized per unit of protein in expressing cell were associated with a weak IFN-γ response. Poor

  13. Critical evaluation of reverse engineering tool Imagix 4D!

    Science.gov (United States)

    Yadav, Rashmi; Patel, Ravindra; Kothari, Abhay

    2016-01-01

    The comprehension of legacy codes is difficult to understand. Various commercial reengineering tools are available that have unique working styles, and are equipped with their inherent capabilities and shortcomings. The focus of the available tools is in visualizing static behavior not the dynamic one. Therefore, it is difficult for people who work in software product maintenance, code understanding reengineering/reverse engineering. Consequently, the need for a comprehensive reengineering/reverse engineering tool arises. We found the usage of Imagix 4D to be good as it generates the maximum pictorial representations in the form of flow charts, flow graphs, class diagrams, metrics and, to a partial extent, dynamic visualizations. We evaluated Imagix 4D with the help of a case study involving a few samples of source code. The behavior of the tool was analyzed on multiple small codes and a large code gcc C parser. Large code evaluation was performed to uncover dead code, unstructured code, and the effect of not including required files at preprocessing level. The utility of Imagix 4D to prepare decision density and complexity metrics for a large code was found to be useful in getting to know how much reengineering is required. At the outset, Imagix 4D offered limitations in dynamic visualizations, flow chart separation (large code) and parsing loops. The outcome of evaluation will eventually help in upgrading Imagix 4D and posed a need of full featured tools in the area of software reengineering/reverse engineering. It will also help the research community, especially those who are interested in the realm of software reengineering tool building.

  14. Structures of Reverse Transcriptase Pre- and Post-Excision Complexes Shed New Light on HIV-1 AZT Resistance

    Directory of Open Access Journals (Sweden)

    Walter A. Scott

    2011-01-01

    Full Text Available HIV-1 resistance to 3'-azido-2',3'-deoxythymidine (AZT, zidovudine results from mutations in reverse transcriptase that increase the ability of the enzyme to excise AZT-monophosphate after it has been incorporated. Crystal structures of complexes of wild type and mutant reverse transcriptase with double-stranded DNA with or without the excision product, AZT adenosine dinucleoside tetraphosphate (AZTppppA, have recently been reported [1]. The excision-enhancing mutations dramatically change the way the enzyme interacts with the excision product.

  15. Structure of HIV-1 Reverse Transcriptase with the Inhibitor -thujaplicinol Bound at the RNase H Active Site

    Energy Technology Data Exchange (ETDEWEB)

    Himmel, D.; Maegley, K; Pauly, T; Bauman, J; Das, K; Dharia, C; Clark, Jr., A; Ryan, K; Hickey, M; et al.

    2009-01-01

    Novel inhibitors are needed to counteract the rapid emergence of drug-resistant HIV variants. HIV-1 reverse transcriptase (RT) has both DNA polymerase and RNase H (RNH) enzymatic activities, but approved drugs that inhibit RT target the polymerase. Inhibitors that act against new targets, such as RNH, should be effective against all of the current drug-resistant variants. Here, we present 2.80 {angstrom} and 2.04 {angstrom} resolution crystal structures of an RNH inhibitor, {beta}-thujaplicinol, bound at the RNH active site of both HIV-1 RT and an isolated RNH domain. {beta}-thujaplicinol chelates two divalent metal ions at the RNH active site. We provide biochemical evidence that {beta}-thujaplicinol is a slow-binding RNH inhibitor with noncompetitive kinetics and suggest that it forms a tropylium ion that interacts favorably with RT and the RNA:DNA substrate.

  16. Inhibition of HIV-1 reverse transcriptase, toxicological and chemical profile of Calophyllum brasiliense extracts from Chiapas, Mexico.

    Science.gov (United States)

    César, García-Zebadúa Julio; Alfonso, Magos-Guerrero Gil; Marius, Mumbrú-Massip; Elizabeth, Estrada-Muñoz; Angel, Contreras-Barrios Miguel; Maira, Huerta-Reyes; Guadalupe, Campos-Lara María; Manuel, Jiménez-Estrada; Ricardo, Reyes-Chilpa

    2011-10-01

    Calophyllum species are sources of calanolides, which inhibit human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT). The hexane extract of the leaves from C. brasiliense collected in Soconusco, State of Chiapas, Mexico, analyzed by HPLC showed to contain apetalic acid, calanolides B, and C. It showed potent anti-HIV-1 RT inhibition (IC(50)=20.2 μg/ml), but was not toxic in mice (LD(50)=1.99 g/kg). The histological study of the mice treated at the highest dose revealed no alteration on hepatocytes, and an increase in the number of spleen megakaryocytes. These results suggest this extract is suitable to continue studies for developing a phytodrug against HIV-1. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Novel bimodular DNA aptamers with guanosine quadruplexes inhibit phylogenetically diverse HIV-1 reverse transcriptases.

    Science.gov (United States)

    Michalowski, Daniel; Chitima-Matsiga, Rebecca; Held, Daniel M; Burke, Donald H

    2008-12-01

    DNA aptamers RT5, RT6 and RT47 form a group of related sequences that inhibit HIV-1 reverse transcriptase (RT). The essential inhibitory structure is identified here as bimodular, with a 5' stem-loop module physically connected to a 3'-guanosine quadruplex module. The stem-loop tolerates considerable sequence plasticity. Connections between the guanosine triplets in the quadruplex could be simplified to a single nucleotide or a nonnucleic acid linker, such as hexaethylene glycol. All 12 quadruplex guanosines are required in an aptamer retaining most of the original loop sequence from RT6; only 11 are required for aptamer R1T (single T residue in intra-quadruplex loops). Circular dichroism (CD) spectroscopy gave ellipticity minima and maxima at 240 nm and 264 nm, indicating a parallel arrangement of the quadruplex strands. The simplified aptamers displayed increased overall stability. An aptamer carrying the original intra-quadruplex loops from RT6 inhibited RT in K(+) buffers but not in Na(+) buffers and displayed significant CD spectral broadening in Na(+) buffers, while R1T inhibited RT in both buffers and displayed less broadening in Na(+) buffers. The bimodular ssDNA aptamers inhibited RT from diverse primate lentiviruses with low nM IC(50) values. These data provide insight into the requirements for broad-spectrum RT inhibition by nucleic acid aptamers.

  18. Dideoxynucleoside HIV reverse transcriptase inhibitors and drug-related hepatotoxicity: a case report

    Directory of Open Access Journals (Sweden)

    Lapadula Giuseppe

    2007-05-01

    Full Text Available Abstract This report regards the case of a 43 year-old HIV-positive woman who developed an episode of serious transaminase elevation during stavudine-including antiretroviral therapy. Diagnostic assessment ruled out hepatitis virus co-infection, alcohol abuse besides other possible causes of liver damage. No signs of lactic acidosis were present. Liver biopsy showed portal inflammatory infiltrate, spotty necrosis, vacuoles of macro- and micro-vesicular steatosis, acidophil and foamy hepatocytes degeneration with organelles clumping, poorly formed Mallory bodies and neutrophil granulocytes attraction (satellitosis. A dramatic improvement in liver function tests occurred when stavudine was discontinued and a new antiretroviral regimen with different nucleoside reverse transcriptase inhibitors was used. The importance of considering hepatotoxicity as an adverse event of HAART including stavudine, even in absence of other signs of mitochondrial toxicity should therefore be underlined. Liver biopsy may provide further important information regarding patients with severe transaminase elevation, for a better understanding of the etiology of liver damage.

  19. COMPUTED TOMOGRAPHIC EVALUATION OF POSTERIOR REVERSIBLE ENCEPHALOPATHY SYNDROME

    Directory of Open Access Journals (Sweden)

    Vishwaprem Raj

    2016-04-01

    Full Text Available BACKGROUND AND PURPOSE Posterior Reversible Encephalopathy Syndrome (PRES is a neurotoxic state that occurs secondary to the inability of posterior circulation to autoregulate. The clinical spectrum and the underlying pathophysiology are still poorly defined. No conclusive evidence has been put forward regarding the relationship between clinical conditions and specific imaging findings of severity or location of oedema. PURPOSE To assess the role of computed tomography in evaluation of Posterior Reversible Encephalopathy Syndrome. MATERIALS AND METHODS 55 patients referred to the Department of Radio-Diagnosis, with a history of neurological abnormalities, including altered mental function, visual loss, stupor with a predisposing history favouring PRES and followed up for a period of 10 – 30 days. RESULTS 21 patients (38.2% were females. 32 patients (58.1% were in the age group between 21 to 30 years. Predisposing condition; 16 (29.1% presented with pre-eclampsia, 12 (21.8% with post-partum status in altered sensorium, 9 (16.4% with seizures, 7 (12.7% with hypertension, 6 (10.9% with visual disturbances, 4 (7.3% with eclampsia and 1 (1.8% with uraemia. 20 cases (36.4% showed findings suggestive of posterior reversible encephalopathy syndrome on initial computed tomography examination. 35 cases showed no initial radiological evidence suggestive of posterior reversible encephalopathy syndrome. Of the 20 cases which showed computed tomographic evidence of posterior reversible encephalopathy syndrome, recovery was noted in 5 cases (9.1%. Persistence of findings detected on first CT was noted in 13 patients (23.6%. Regional predominance of the lesions was as follows. Frontal lobe (39%, Parietal lobe (32%, Temporal lobe (15% and occipital lobe (15%. CONCLUSION Varied clinical manifestations are associated with anatomical findings recognisable by neuro-imaging as PRES. Prompt imaging is necessary for the recognition of the condition and appropriate

  20. A Polymorphism at Position 400 in the Connection Subdomain of HIV-1 Reverse Transcriptase Affects Sensitivity to NNRTIs and RNaseH Activity

    NARCIS (Netherlands)

    D.J. Wright (David Justin); I. Deuzing (Ilona); P. Flandre (Philippe); P. van den Eede (Peter); M. Govaert (Micheline); L. Setiawan (Laurentia); P.V. Coveney (Peter); A.-G. Marcelin (Anne-Geneviève); V. Calvez (Vincent); C.A. Boucher (Charles); N. Beerens (Nancy)

    2013-01-01

    textabstractReverse transcriptase (RT) plays an essential role in HIV-1 replication, and inhibition of this enzyme is a key component of HIV-treatment. However, the use of RT inhibitors can lead to the emergence of drug-resistant variants. Until recently, most clinically relevant resistance

  1. Multi-nucleoside reverse transcriptase inhibitor resistant HIV type-1 in a patient from Sierra Leone failing stavudine, lamivudine and nevirapine

    NARCIS (Netherlands)

    Hamers, Raph L.; Wensing, Annemarie M. J.; Back, Nicole K. T.; Arcilla, Maria S.; Frissen, Jos P. H.

    2011-01-01

    We report a 33-year-old HIV type-1 (HIV-1)-infected male from Sierra Leone who harboured extensive drug resistance mutations to all nucleoside reverse transcriptase inhibitors (NRTIs) and non-NRTIs, including the multi-NRTI-resistance Q151M complex, K65R, M184I and Y181I, after using standard

  2. Research on the Environmental Performance Evaluation of Electronic Waste Reverse Logistics Enterprise

    Science.gov (United States)

    Yang, Yu-Xiang; Chen, Fei-Yang; Tong, Tong

    According to the characteristic of e-waste reverse logistics, environmental performance evaluation system of electronic waste reverse logistics enterprise is proposed. We use fuzzy analytic hierarchy process method to evaluate the system. In addition, this paper analyzes the enterprise X, as an example, to discuss the evaluation method. It's important to point out attributes and indexes which should be strengthen during the process of ewaste reverse logistics and provide guidance suggestions to domestic e-waste reverse logistics enterprises.

  3. Efficient in vitro inhibition of HIV-1 gag reverse transcription by peptide nucleic acid (PNA) at minimal ratios of PNA/RNA

    DEFF Research Database (Denmark)

    Koppelhus, Uffe; Zachar, Vladimir; Nielsen, P.E.

    1997-01-01

    We have tested the inhibitory potential of peptide nucleic acid (PNA) on in vitro reverse transcription of the HIV-1 gag gene. PNA was designed to target different regions of the HIV-1 gag gene and the effect on reverse transcription by HIV-1, MMLV and AMV reverse transcriptases (RTs...... that would indicate PNA-mediated RNase H activation of the tested RTs. In conclusion, PNA appears to have a potential to become a specific and efficient inhibitor of reverse transcription in vivo , provided sufficient intracellular levels are achievable.......) was investigated. We found that a bis-PNA (parallel antisense 10mer linked to antiparallel antisense 10mer) was superior to both the parallel antisense 10mer and antiparallel antisense 10mer in inhibiting reverse transcription of the gene, thus indicating triplex formation at the target sequence. A complete arrest...

  4. HIV-1 Reverse Transcriptase Structure with RNase H Inhibitor dihydroxy benzoyl naphthyl Hydrazone Bound at a Novel Site

    Energy Technology Data Exchange (ETDEWEB)

    Himmel,D.; Sarafianos, S.; Dharmasena, S.; Hossain, M.; McCoy-Simandle, K.; Ilina, T.; Clark, A.; Knight, J.; Julias, J.; et al.

    2007-01-01

    The rapid emergence of drug-resistant variants of human immunodeficiency virus, type 1 (HIV-1), has limited the efficacy of anti-acquired immune deficiency syndrome (AIDS) treatments, and new lead compounds that target novel binding sites are needed. We have determined the 3.15 {angstrom} resolution crystal structure of HIV-1 reverse transcriptase (RT) complexed with dihydroxy benzoyl naphthyl hydrazone (DHBNH), an HIV-1 RT RNase H (RNH) inhibitor (RNHI). DHBNH is effective against a variety of drug-resistant HIV-1 RT mutants. While DHBNH has little effect on most aspects of RT-catalyzed DNA synthesis, at relatively high concentrations it does inhibit the initiation of RNA-primed DNA synthesis. Although primarily an RNHI, DHBNH binds >50 {angstrom} away from the RNH active site, at a novel site near both the polymerase active site and the non-nucleoside RT inhibitor (NNRTI) binding pocket. When DHBNH binds, both Tyr181 and Tyr188 remain in the conformations seen in unliganded HIV-1 RT. DHBNH interacts with conserved residues (Asp186, Trp229) and has substantial interactions with the backbones of several less well-conserved residues. On the basis of this structure, we designed substituted DHBNH derivatives that interact with the NNRTI-binding pocket. These compounds inhibit both the polymerase and RNH activities of RT.

  5. Evaluation of reversible contraceptive potential of Cordia dichotoma leaves extract

    Directory of Open Access Journals (Sweden)

    Plaban Bhattacharya

    2013-03-01

    Full Text Available Considering the safety-risk ratio of steroidal contraceptives, the present work was carried out to evaluate ethno-contraceptive use of Cordia dichotoma G. Forst., Boraginaceae, leaves (LCD. Preliminary pharmacological screening was performed on post-coital female albino rats. The leaves extract (LD50 5.50 g/kg bw showed 100% anti-implantation activity (n=10 at 800 mg/kg dose level. (2-hydroxypropyl-β-cyclodextrin (BCD was used as bioavailability enhancer to form LCD-BCD complex, characterized by DLS, SEM and XRD analyses. The LCD-BCD complex (1:1, w/w exhibited 100% pregnancy interception (n=20 at the dose level of 250 mg/kg and also showed strong estrogenic potential with a luteal phase defect. Qualitative and quantitative phytochemical analyses were carried out. The LCD extract was standardized by a validated HPTLC method and two contraceptive phytoconstituents, apigenin and luteolin were isolated. A detailed pharmacological analyses followed by chronic toxicity study were performed to predict the reversible nature of the developed phytopharmaceutical. The histological and biochemical estimations detected the reversible contraceptive potential after withdrawal. The observations suggested that the developed phyto-pharmaceutical has potential antifertility activity with safety aspects.

  6. Evaluation of reversible contraceptive potential of Cordia dichotoma leaves extract

    Directory of Open Access Journals (Sweden)

    Plaban Bhattacharya

    2013-04-01

    Full Text Available Considering the safety-risk ratio of steroidal contraceptives, the present work was carried out to evaluate ethno-contraceptive use of Cordia dichotoma G. Forst., Boraginaceae, leaves (LCD. Preliminary pharmacological screening was performed on post-coital female albino rats. The leaves extract (LD50 5.50 g/kg bw showed 100% anti-implantation activity (n=10 at 800 mg/kg dose level. (2-hydroxypropyl-β-cyclodextrin (BCD was used as bioavailability enhancer to form LCD-BCD complex, characterized by DLS, SEM and XRD analyses. The LCD-BCD complex (1:1, w/w exhibited 100% pregnancy interception (n=20 at the dose level of 250 mg/kg and also showed strong estrogenic potential with a luteal phase defect. Qualitative and quantitative phytochemical analyses were carried out. The LCD extract was standardized by a validated HPTLC method and two contraceptive phytoconstituents, apigenin and luteolin were isolated. A detailed pharmacological analyses followed by chronic toxicity study were performed to predict the reversible nature of the developed phytopharmaceutical. The histological and biochemical estimations detected the reversible contraceptive potential after withdrawal. The observations suggested that the developed phyto-pharmaceutical has potential antifertility activity with safety aspects.

  7. An evaluation of the SD Bioline HIV/syphilis duo test.

    Science.gov (United States)

    Holden, Jeffrey; Goheen, Joshua; Jett-Goheen, Mary; Barnes, Mathilda; Hsieh, Yu-Hsiang; Gaydos, Charlotte A

    2018-01-01

    Many health agencies now recommend routine HIV and syphilis testing for pregnant women and most-at-risk populations such as men who have sex with men. With the increased availability of highly sensitive, low cost rapid point-of-care tests, the ability to meet those recommendations has increased, granting wider access to quick and accurate diagnoses. Using blood specimens collected from a Baltimore City Health Department (BCHD) sexually transmitted infection clinic, we evaluated the SD Bioline HIV/Syphilis Duo, a rapid test that simultaneously detects antibodies to HIV and syphilis and has the potential to further benefit clinics and patients by reducing costs, testing complexity, and patient wait times. SD DUO HIV sensitivity and specificity, when compared to BCHD results, were 91.7 and 99.5%, respectively. SD DUO syphilis sensitivity and specificity, when compared to rapid plasma reagin, were 85.7 and 96.8%, respectively, and 69.7 and 99.7%, respectively, when compared to Treponema pallidum particle agglutination (TPPA). SD DUO syphilis sensitivity and specificity, when compared to a traditional screening algorithm, improved to 92.3 and 100%, respectively, and improved to 72.9 and 99.7%, respectively, when compared to a reverse screening algorithm. The HIV component of the SD DUO performed moderately well. However, results for the SD DUO syphilis component, when compared to TPPA, support the need for further testing and assessment.

  8. Evaluation of Olfactory and Gustatory Function of HIV Infected Women

    Directory of Open Access Journals (Sweden)

    Ayotunde James Fasunla

    2016-01-01

    Full Text Available Background. Compliance with medication requires good sense of smell and taste. Objective. To evaluate the olfactory and gustatory function of HIV infected women in Ibadan, Nigeria. Methods. A case control study of women comprising 83 HIV infected women and 79 HIV uninfected women. Subjective self-rating of taste and smell function was by visual analogue scale. Olfactory function was measured via olfactory threshold (OT, olfactory discrimination (OD, olfactory identification (OI, and TDI using “Sniffin’ sticks” kits and taste function (Total Taste Strips (TTS score measurement was by taste strips. Results. The mean age of the HIV infected women was 43.67 years ± 10.72 and control was 41.48 years ± 10.99. There was no significant difference in the self-reported assessment of smell (p=0.67 and taste (p=0.84 of HIV infected and uninfected women. Although the mean OT, OD, OI, TDI, and TTS scores of HIV infected and uninfected women were within the normosmic and normogeusic values, the values were significantly higher in the controls (p<0.05. Hyposmia was in 39.7% of subjects and 12.6% of controls while hypogeusia was in 15.7% of subjects and 1.3% of controls. Conclusions. Hyposmia and hypogeusia are commoner among the HIV infected women than the HIV uninfected women and the risk increases with an increased duration of highly active antiretroviral therapy.

  9. A Novel Bromodomain Inhibitor Reverses HIV-1 Latency through Specific Binding with BRD4 to Promote Tat and P-TEFb Association

    Directory of Open Access Journals (Sweden)

    Huachao Huang

    2017-06-01

    Full Text Available While combinatory antiretroviral therapy (cART can effectively reduce HIV-1 viremia, it cannot eliminate HIV-1 infection. In the presence of cART, viral reservoirs remain latent, impeding the cure of HIV-1/AIDS. Recently, latency-reversing agents (LRAs have been developed with the intent of purging latent HIV-1, providing an intriguing strategy for the eradication of the residual viral reservoirs. Our earlier studies show that the first-generation, methyl-triazolo bromodomain, and extra-terminal domain inhibitor (BETi, JQ1, facilitates the reversal of HIV-1 latency. BETis have emerged as a new class of compounds that are promising for this HIV-1 “shock and kill” eradication approach. However, when used as a single drug, JQ1 only modestly reverses HIV-1 latency, which complicates studying the underlining mechanisms. Meanwhile, it has been widely discussed that the induction of latent proviruses is stochastic (Ho et al., 2013. Thus, new BETis are currently under active development with focus on improving potency, ease of synthesis and structural diversity. Using fluorous-tagged multicomponent reactions, we developed a novel second-generation, 3,5-dimethylisoxazole BETi based on an imidazo[1,2-a] pyrazine scaffold, UMB-32. Furthermore, we screened 37 UMB-32 derivatives and identified that one, UMB-136, reactivates HIV-1 in multiple cell models of HIV-1 latency with better efficiency than either JQ1 or UMB-32. UMB-136 enhances HIV-1 transcription and increases viral production through the release of P-TEFb. Importantly, UMB-136 enhances the latency-reversing effects of PKC agonists (prostratin, bryostatin-1 in CD8-depleted PBMCs containing latent viral reservoirs. Our results illustrate that structurally improved BETis, such as UMB-136, may be useful as promising LRAs for HIV-1 eradication.

  10. Altered error specificity of RNase H-deficient HIV-1 reverse transcriptases during DNA-dependent DNA synthesis

    Science.gov (United States)

    Álvarez, Mar; Barrioluengo, Verónica; Afonso-Lehmann, Raquel N.; Menéndez-Arias, Luis

    2013-01-01

    Asp443 and Glu478 are essential active site residues in the RNase H domain of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT). We have investigated the effects of substituting Asn for Asp443 or Gln for Glu478 on the fidelity of DNA-dependent DNA synthesis of phylogenetically diverse HIV-1 RTs. In M13mp2 lacZα-based forward mutation assays, HIV-1 group M (BH10) and group O RTs bearing substitutions D443N, E478Q, V75I/D443N or V75I/E478Q showed 2.0- to 6.6-fold increased accuracy in comparison with the corresponding wild-type enzymes. This was a consequence of their lower base substitution error rates. One-nucleotide deletions and insertions represented between 30 and 68% of all errors identified in the mutational spectra of RNase H-deficient HIV-1 group O RTs. In comparison with the wild-type RT, these enzymes showed higher frameshift error rates and higher dissociation rate constants (koff) for DNA/DNA template–primers. The effects on frameshift fidelity were similar to those reported for mutation E89G and suggest that in HIV-1 group O RT, RNase H inactivation could affect template/primer slippage. Our results support a role for the RNase H domain during plus-strand DNA polymerization and suggest that mutations affecting RNase H function could also contribute to retrovirus variability during the later steps of reverse transcription. PMID:23444139

  11. Identification of a Novel Scaffold for Allosteric Inhibition of Wild Type and Drug Resistant HIV-1 Reverse Transcriptase by Fragment Library Screening

    NARCIS (Netherlands)

    Geitmann, M.; Elinder, M; Seeger, C; Brandt, P; de Esch, I.J.P.; Danielson, U.H.

    2011-01-01

    A novel scaffold inhibiting wild type and drug resistant variants of human immunodeficiency virus type 1 reverse transcriptase (HIV-1RT) has been identified in a library consisting of 1040 fragments. The fragments were significantly different from already known non-nucleoside reverse transcriptase

  12. Evaluation of the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test as an alternative to Western blot for confirmation of HIV infection.

    Science.gov (United States)

    Cárdenas, Ana María; Baughan, Eleonore; Hodinka, Richard L

    2013-12-01

    In the United States, a new HIV diagnostic algorithm has been proposed that uses an HIV-1/HIV-2 antibody differentiation immunoassay instead of Western blot or immunofluoresence for confirmatory testing. To evaluate the Multispot HIV-1/HIV-2 Rapid Test (Multispot) as an alternative to Western blot analysis for confirmation of HIV infection. A series of 205 serum and plasma specimens positive for HIV-1 or HIV-2 were used to compare the performance of Multispot to a standard HIV-1 Western blot. Positive samples included 63 specimens from patients>18 months of age, 33 proficiency survey specimens, and 109 specimens from nine commercial seroconversion and performance panels. In addition, 63 specimens from 51 HIV-exposed, uninfected children≤18 months of age in various stages of seroreversion and 192 HIV-negative samples were tested. Specimens were initially screened using a 4th generation HIV Ag/Ab Combo assay. Multispot readily discriminated between individuals with HIV-1 or HIV-2 infection and those who were uninfected. Of the 205 samples repeatedly reactive by the 4th generation screening assay, infection status was correctly confirmed by Multispot in 83.9% (172/205) compared to 68.8% (141/205) for Western blot. Multispot detected HIV-1 earlier in 27.6% of low-titer antibody specimens called indeterminate by Western blot, and effectively reduced the number of indeterminate results in seroreverting HIV-1 exposed, uninfected infants and for HIV-2 infections misinterpreted as indeterminate or positive by HIV-1 Western blot. Multispot offers speed and simplicity over Western blot and has an excellent performance for differentiation and confirmation of antibodies to HIV-1 and HIV-2. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. A simian-human immunodeficiency virus carrying the rt gene from Chinese CRF01_AE strain of HIV is sensitive to nucleoside reverse transcriptase inhibitors and has a highly genetic stability in vivo.

    Science.gov (United States)

    Wang, Wei; Yao, Nan; Ju, Bin; Dong, Zhihui; Cong, Zhe; Jiang, Hong; Qin, Chuan; Wei, Qiang

    2014-06-01

    Human immunodeficiency virus (HIV)-1 subtype CRF01_AE is one of the major HIV-1 subtypes that dominate the global epidemic. However, its drug resistance, associated mutations, and viral fitness have not been systemically studied, because available chimeric simian-HIVs (SHIVs) usually express the HIV-1 reverse transcriptase (rt) gene of subtype B HIV-1, which is different from subtype CRF01_AE HIV-1. In this study, a recombinant plasmid, pRT-SHIV/AE, was constructed to generate a chimeric RT-SHIV/AE by replacing the rt gene of simian immunodeficiency virus (SIVmac239) with the counterpart of Chinese HIV-1 subtype CRF01_AE. The infectivity, replication capacity, co-receptor tropism, drug sensitivity, and genetic stability of RT-SHIV/AE were characterized. The new chimeric RT-SHIV/AE effectively infected and replicated in human T cell line and rhesus peripheral blood mononuclear cells (rhPBMC). The rt gene of RT-SHIV/AE lacked the common mutation (T215I) associated with drug resistance. RT-SHIV-AE retained infectivity and immunogenicity, similar to that of its counterpart RT-SHIV/TC virus following intravenous inoculation in Chinese rhesus macaque. RT-SHIV-AE was more sensitive to nucleoside reverse transcriptase inhibitors (NRTIs) than the RT-SHIV/TC. RT-SHIV/AE was genetically stable in Chinese rhesus macaque. The new chimeric RT-SHIV/AE may be a valuable tool for evaluating the efficacy of the rt-based antiviral drugs against the subtype CRF01_AE HIV-1. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  14. Evaluation of nine HIV rapid test kits to develop a national HIV testing algorithm in Nigeria

    Directory of Open Access Journals (Sweden)

    Orji Bassey

    2015-05-01

    Full Text Available Background: Non-cold chain-dependent HIV rapid testing has been adopted in many resource-constrained nations as a strategy for reaching out to populations. HIV rapid test kits (RTKs have the advantage of ease of use, low operational cost and short turnaround times. Before 2005, different RTKs had been used in Nigeria without formal evaluation. Between 2005 and 2007, a study was conducted to formally evaluate a number of RTKs and construct HIV testing algorithms. Objectives: The objectives of this study were to assess and select HIV RTKs and develop national testing algorithms. Method: Nine RTKs were evaluated using 528 well-characterised plasma samples. These comprised 198 HIV-positive specimens (37.5% and 330 HIV-negative specimens (62.5%, collected nationally. Sensitivity and specificity were calculated with 95% confidence intervals for all nine RTKs singly and for serial and parallel combinations of six RTKs; and relative costs were estimated. Results: Six of the nine RTKs met the selection criteria, including minimum sensitivity and specificity (both ≥ 99.0% requirements. There were no significant differences in sensitivities or specificities of RTKs in the serial and parallel algorithms, but the cost of RTKs in parallel algorithms was twice that in serial algorithms. Consequently, three serial algorithms, comprising four test kits (BundiTM, DetermineTM, Stat-Pak® and Uni-GoldTM with 100.0% sensitivity and 99.1% – 100.0% specificity, were recommended and adopted as national interim testing algorithms in 2007. Conclusion: This evaluation provides the first evidence for reliable combinations of RTKs for HIV testing in Nigeria. However, these RTKs need further evaluation in the field (Phase II to re-validate their performance.

  15. Proteochemometric modeling of the susceptibility of mutated variants of the HIV-1 virus to reverse transcriptase inhibitors.

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    Muhammad Junaid

    Full Text Available BACKGROUND: Reverse transcriptase is a major drug target in highly active antiretroviral therapy (HAART against HIV, which typically comprises two nucleoside/nucleotide analog reverse transcriptase (RT inhibitors (NRTIs in combination with a non-nucleoside RT inhibitor or a protease inhibitor. Unfortunately, HIV is capable of escaping the therapy by mutating into drug-resistant variants. Computational models that correlate HIV drug susceptibilities to the virus genotype and to drug molecular properties might facilitate selection of improved combination treatment regimens. METHODOLOGY/PRINCIPAL FINDINGS: We applied our earlier developed proteochemometric modeling technology to analyze HIV mutant susceptibility to the eight clinically approved NRTIs. The data set used covered 728 virus variants genotyped for 240 sequence residues of the DNA polymerase domain of the RT; 165 of these residues contained mutations; totally the data-set covered susceptibility data for 4,495 inhibitor-RT combinations. Inhibitors and RT sequences were represented numerically by 3D-structural and physicochemical property descriptors, respectively. The two sets of descriptors and their derived cross-terms were correlated to the susceptibility data by partial least-squares projections to latent structures. The model identified more than ten frequently occurring mutations, each conferring more than two-fold loss of susceptibility for one or several NRTIs. The most deleterious mutations were K65R, Q151M, M184V/I, and T215Y/F, each of them decreasing susceptibility to most of the NRTIs. The predictive ability of the model was estimated by cross-validation and by external predictions for new HIV variants; both procedures showed very high correlation between the predicted and actual susceptibility values (Q2=0.89 and Q2ext=0.86. The model is available at www.hivdrc.org as a free web service for the prediction of the susceptibility to any of the clinically used NRTIs for any HIV

  16. Evaluation of the siemens HIV antigen-antibody immunoassay.

    Science.gov (United States)

    Vallefuoco, Luca; Aden Abdi, Fatima; Sorrentino, Rosanna; Spalletti-Cernia, Daniela; Mazzarella, Claudia; Barbato, Sara; Perna, Enzo; Buffolano, Wilma; Di Nicuolo, Giuseppe; Portella, Giuseppe

    2014-01-01

    Fourth-generation assays for the simultaneous detection of human immunodeficiency virus (HIV) antigen and antibodies are available on the international market and are currently used for blood donor screening and for HIV diagnosis. In this study we evaluated the performance of the novel automated fourth-generation ADVIA Centaur® HIV Ag/Ab Combo assay. The assay detected seroconversion at the same bleed or at least one bleed earlier in panels with respect to other assays and showed a detection efficacy equal to those of other assays in a low-titer panel. Samples obtained from blood donors (n = 2,778) or from HIV-positive patients (HIV-1 B subtype, n = 82; non-B subtype, n = 71) were also tested, showing a good correlation with other fourth-generation assays. We assessed the performance of 3 fourth-generation assays for detecting in utero transmitted anti-HIV antibodies and found a more specific detection efficiency with the ADVIA Centaur HIV Ag/Ab Combo assay compared to the other fourth-generation assays.

  17. Evaluation of four rapid tests for diagnosis and differentiation of HIV-1 and HIV-2 infections in Guinea-Conakry, West Africa.

    OpenAIRE

    Chaillet, Pascale; Tayler-Smith, Katie; Zachariah, Rony; Duclos, Nanfack; Moctar, Diallo; Beelaert, Greet; Fransen, Katrien

    2010-01-01

    With both HIV-1 and HV-2 prevalent in Guinea-Conakry, accurate diagnosis and differentiation is crucial for treatment purposes. Thus, four rapid HIV tests were evaluated for their HIV-1 and HIV-2 diagnostic and discriminative capacity for use in Guinea-Conakry. These included SD Bioline HIV 1/2 3.0 (Standard Diagnostics Inc.), Genie II HIV1/HIV2 (Bio-Rad), First Response HIV Card Test 1-2.0 (PMC Medical) and Immunoflow HIV1-HIV2 (Core Diagnostics). Results were compared with gold standard tes...

  18. HIV reverse transcriptase inhibiting antibodies detected by a new technique: relation to p24 and gp41 antibodies, HIV antigenemia and clinical variables.

    Science.gov (United States)

    Neumüller, M; Karlsson, A; Lennerstrand, J; Källander, C F; Holmberg, V; Långström-Persson, U; Thorstensson, R; Sandström, E; Gronowitz, J S

    1991-05-01

    A new assay for HIV reverse transcriptase activity inhibiting antibodies (RTI-ab) was used for the analysis of a large collection of sera sampled before and after confirmation of HIV infection. In this assay HIV-RT was preincubated with diluted serum, after which residual RT activity was determined by a technique using a template coupled to macrobeads and 125I-lodo-deoxyuridine-triphosphate as the tracer-substrate. Of the 936 sera analysed, 818 were found positive for RTI-ab, and 824 were positive in Western blot (Wb). The prevalence of RTI-ab compared to Wb was therefore 99.3%. The corresponding figure for 930 sera analysed for envelope-ab, i.e., gp41-ab, was 823 positive, and of these 930 sera 815 were Wb positive, giving a comparative prevalence of 101%. In contrast, only 678 samples of 993 analyzed for core ab, i.e., p24, were positive, giving a prevalence of 77.0% as 880 of these samples were Wb positive. Thus, RTI-ab was as prevalent as gp41-ab, and although the analyses of RTI-ab amounts in different stages showed decreasing levels in stage IV compared to stages II or III, all of the sera except 1 were found positive in stages III and IV. Further, it was found that both the few RTI-ab negative samples in stage II and the few RTI-ab positive samples among Wb negative sera were sampled in connection with seroconversion. The specificity of the RTI-ab assay was 100% in a test of 200 serum samples from HIV negative blood donors. It was concluded that RTI-ab analyses can be made highly sensitive and specific and useful for studies of HIV infection.

  19. Identification of a 3-aminoimidazo[1,2-a]pyridine inhibitor of HIV-1 reverse transcriptase

    Directory of Open Access Journals (Sweden)

    Elleder Daniel

    2012-12-01

    Full Text Available Abstract Background Despite the effectiveness of highly active antiretroviral therapy (HAART, there remains an urgent need to develop new human immunodeficiency virus type 1 (HIV-1 inhibitors with better pharmacokinetic properties that are well tolerated, and that block common drug resistant virus strains. Methods Here we screened an in-house small molecule library for novel inhibitors of HIV-1 replication. Results An active compound containing a 3-aminoimidazo[1,2-a]pyridine scaffold was identified and quantitatively characterized as a non-nucleoside reverse transcriptase inhibitor (NNRTI. Conclusions The potency of this compound coupled with its inexpensive chemical synthesis and tractability for downstream SAR analysis make this inhibitor a suitable lead candidate for further development as an antiviral drug.

  20. An economic evaluation of mandatory premarital testing for HIV.

    Science.gov (United States)

    McKay, N L; Phillips, K M

    1991-01-01

    This study provides a comprehensive assessment of mandatory premarital testing for HIV. We incorporate new evidence about the assumptions underlying an economic evaluation of premarital testing, use standard methods of program evaluation (cost-effectiveness analysis and cost-benefit analysis), and carefully evaluate how changes in the various assumptions affect the results. The cost-effectiveness results show that, under the most likely conditions, the cost per case of HIV infection prevented by mandatory premarital testing would be between $70,000 and $127,000. In the cost-benefit analysis, the benefit-cost ratio in the most likely scenarios ranges between 3.1 and 28.2.

  1. Enhanced activity of carbosilane dendrimers against HIV when combined with reverse transcriptase inhibitor drugs: searching for more potent microbicides

    Science.gov (United States)

    Vacas-Córdoba, Enrique; Galán, Marta; de la Mata, Francisco J; Gómez, Rafael; Pion, Marjorie; Muñoz-Fernández, M Ángeles

    2014-01-01

    Self-administered topical microbicides or oral preexposure prophylaxis could be very helpful tools for all risk groups to decrease the human immunodeficiency virus (HIV)-1 infection rates. Up until now, antiretrovirals (ARVs) have been the most advanced microbicide candidates. Nevertheless, the majority of clinical trials has failed in HIV-1 patients. Nanotechnology offers suitable approaches to develop novel antiviral agents. Thereby, new nanosystems, such as carbosilane dendrimers, have been shown to be safe and effective compounds against HIV with great potential as topical microbicides. In addition, because most of the attempts to develop effective topical microbicides were unsuccessful, combinatorial strategies could be a valid approach when designing new microbicides. We evaluated various combinations of anionic carbosilane dendrimers with sulfated (G3-S16) and naphthyl sulfonated (G2-NF16) ended groups with different ARVs against HIV-1 infection. The G3-S16 and G2-NF16 dendrimers showed a synergistic or additive activity profile with zidovudine, efavirenz, and tenofovir in the majority of the combinations tested against the X4 and R5 tropic HIV-1 in cell lines, as well as in human primary cells. Therefore, the combination of ARVs and polyanionic carbosilane dendrimers enhances the antiviral potency of the individual compounds, and our findings support further clinical research on combinational approaches as potential microbicides to block the sexual transmission of HIV-1. PMID:25114528

  2. The inophyllums, novel inhibitors of HIV-1 reverse transcriptase isolated from the Malaysian tree, Calophyllum inophyllum Linn.

    Science.gov (United States)

    Patil, A D; Freyer, A J; Eggleston, D S; Haltiwanger, R C; Bean, M F; Taylor, P B; Caranfa, M J; Breen, A L; Bartus, H R; Johnson, R K

    1993-12-24

    As part of a search for novel inhibitors of HIV-1 reverse transcriptase, the acetone extract of the giant African snail, Achatina fulica, was shown to be active. Fractionation of the extract yielded inophyllums A, B, C, and E and calophyllolide (1a, 2a, 3a, 3b, and 6), previously isolated from Calophyllum inophyllum Linn., a known source of nutrition for A. fulica. From a methanol/methylene chloride extract of C. inophyllum, the same natural products in considerably greater yield were isolated in addition to a novel enantiomer of soulattrolide (4), inophyllum P (2b), and two other novel compounds, inophyllums G-1 (7) and G-2 (8). The absolute stereochemistry of inophyllum A (1a) was determined to be 10(R), 11(S), 12(S) from a single-crystal X-ray analysis of its 4-bromobenzoate derivative, and the relative stereochemistries of the other inophyllums isolated from C. inophyllum were established by a comparison of their 1H NMR NOE values and coupling constants to those of inophyllum A (1a). Inophyllums B and P (2a and 2b) inhibited HIV reverse transcriptase with IC50 values of 38 and 130 nM, respectively, and both were active against HIV-1 in cell culture (IC50 of 1.4 and 1.6 microM). Closely related inophyllums A, C, D, and E, including calophyllic acids, were significantly less active or totally inactive, indicating certain structural requirements in the chromanol ring. Altogether, 11 compounds of the inophyllum class were isolated from C. inophyllum and are described together with the SAR of these novel anti-HIV compounds.

  3. Subunit-specific mutational analysis of residue N348 in HIV-1 reverse transcriptase

    Directory of Open Access Journals (Sweden)

    Radzio Jessica

    2011-08-01

    Full Text Available Abstract Background N348I in HIV-1 reverse transcriptase (RT confers resistance to zidovudine (AZT and nevirapine. Biochemical studies demonstrated that N348I indirectly increases AZT resistance by decreasing the frequency of secondary ribonuclease H (RNase H cleavages that reduce the RNA/DNA duplex length of the template/primer (T/P and diminish the efficiency of AZT-monophosphate (MP excision. By contrast, there is some discrepancy in the literature in regard to the mechanisms associated with nevirapine resistance: one study suggested that it is due to decreased inhibitor binding while others suggest that it may be related to the decreased RNase H cleavage phenotype. From a structural perspective, N348 in both subunits of RT resides distal to the enzyme's active sites, to the T/P binding tract and to the nevirapine-binding pocket. As such, the structural mechanisms associated with the resistance phenotypes are not known. Results Using a novel modelled structure of RT in complex with an RNA/DNA T/P, we identified a putative interaction between the β14-β15 loop in the p51 subunit of RT and the RNA template. Substitution of the asparagine at codon 348 in the p51 subunit with either isoleucine or leucine abrogated the observed protein-RNA interaction, thus, providing a possible explanation for the decreased RNase H phenotype. By contrast, alanine or glutamine substitutions exerted no effect. To validate this model, we introduced the N348I, N348L, N348A and N348Q mutations into RT and purified enzymes that contained subunit-specific mutations. N348I and N348L significantly decreased the frequency of secondary RNase H cleavages and increased the enzyme's ability to excise AZT-MP. As predicted by the modelling, this phenotype was due to the mutation in the p51 subunit of RT. By contrast, the N348A and N348Q RTs exhibited RNase H cleavage profiles and AZT-MP excision activities similar to the wild-type enzyme. All N348 mutant RTs exhibited decreased

  4. Substrate mimicry: HIV-1 reverse transcriptase recognizes 6-modified-3'-azido-2',3'-dideoxyguanosine-5'-triphosphates as adenosine analogs.

    Science.gov (United States)

    Herman, Brian D; Schinazi, Raymond F; Zhang, Hong-wang; Nettles, James H; Stanton, Richard; Detorio, Mervi; Obikhod, Aleksandr; Pradère, Ugo; Coats, Steven J; Mellors, John W; Sluis-Cremer, Nicolas

    2012-01-01

    β-D-3'-Azido-2',3'-dideoxyguanosine (3'-azido-ddG) is a potent inhibitor of HIV-1 replication with a superior resistance profile to zidovudine. Recently, we identified five novel 6-modified-3'-azido-ddG analogs that exhibit similar or superior anti-HIV-1 activity compared to 3'-azido-ddG in primary cells. To gain insight into their structure-activity-resistance relationships, we synthesized their triphosphate (TP) forms and assessed their ability to inhibit HIV-1 reverse transcriptase (RT). Steady-state and pre-steady-state kinetic experiments show that the 6-modified-3'-azido-ddGTP analogs act as adenosine rather than guanosine mimetics in DNA synthesis reactions. The order of potency of the TP analogs against wild-type RT was: 3'-azido-2,6-diaminopurine >3'-azido-6-chloropurine; 3'-azido-6-N-allylaminopurine > 2-amino-6-N,N-dimethylaminopurine; 2-amino-6-methoxypurine. Molecular modeling studies reveal unique hydrogen-bonding interactions between the nucleotide analogs and the template thymine base in the active site of RT. Surprisingly, the structure-activity relationship of the analogs differed in HIV-1 RT ATP-mediated excision assays of their monophosphate forms, suggesting that it may be possible to rationally design a modified base analog that is efficiently incorporated by RT but serves as a poor substrate for ATP-mediated excision reactions. Overall, these studies identify a promising strategy to design novel nucleoside analogs that exert profound antiviral activity against both WT and drug-resistant HIV-1.

  5. Evaluation of the acerola juice concentrated by reverse osmosis

    Directory of Open Access Journals (Sweden)

    Eliane Rodrigues dos Santos Gomes

    2005-06-01

    Full Text Available The aim of this study was to obtain the acerola juice using separation processes with membranes. The acerola pulp was initially defrosted and treated with 100 ppm of Citrozym Ultra L enzyme at 45ºC for one hour, then ultrafiltrated at 3 bar at 45ºC using 0.1 µm ceramic membrane, and concentrated by reverse osmosis using a spiral membrane of a compound film. The pressures on the reverse osmosis were 20, 30, and 40 bar at environmental temperature, thus, resulting a juice with 9.76, 14.56, and 17.36 ºBrix, respectively. The physicochemical characteristics of the juice were preserved and, studies on evaluation of the public acceptability, showed that 75% of the consumers liked the juice.O objetivo deste trabalho foi à obtenção de suco de acerola utilizando processos de separação com membranas. Combinou-se a ultrafiltração e a osmose inversa, visando a melhoria do processo produtivo, utilizando-se uma tecnologia limpa. Para a acerola, visou-se manter e concentrar significativamente seu teor de vitamina C, obtendo-se um suco com sabor agradável, o mais próximo possível do suco in natura. A polpa de acerola foi inicialmente descongelada e tratada com 100 ppm da enzima Citrozym Ultra L, à 45º C por 1 hora e posteriormente ultrafiltrada a 3 bar na mesma temperatura em membrana cerâmica de 0,1 µm e na seqüência, concentrada por osmose inversa utilizando membrana espiral de filme composto. As pressões na osmose inversa foram 20, 30 e 40 bar em temperatura ambiente, obtendo-se um suco com 9,76, 14,56 e 17,36 ºBrix respectivamente. As características físico-químicas foram preservadas e na avaliação da aceitabilidade, 75% dos consumidores gostaram do suco, indicando boa aceitação.

  6. Performance evaluation of a new fourth-generation HIV Ag/Ab combination electrochemiluminescence immunoassay - evaluation of a new HIV assay.

    Science.gov (United States)

    Wang, Tingting; Li, Dongdong; Yan, Kening; Yuan, Yu; Yang, Tingfu; Du, Xiaoqing; Yan, Xuedan; Tao, Chuanmin; Wang, Lanlan

    2014-03-01

    A new fourth-generation HIV Ag/Ab electrochemiluminescence immunoassay for screening of HIV infection, the Elecsys HIV Combi PT (Roche Diagnostics, Penzberg, Germany) assay, is going to be commercially available in clinical laboratories in China. This assay was evaluated and compared with two commonly used assays: Elecsys HIV Combi assay and the Livzon anti-HIV-1/2 ELISA. Commercially available panels and 30 established HIV infection samples were tested to evaluate the sensitivity. In addition, a total of 675 routine clinical samples were collected and tested in West China Hospital to compare the specificity. Any reactive result from a screening test was retested and all reactive retested samples were confirmed with Western blot assay, Elecsys HIV Ag test, Elecsys HIV Ag confirmatory test or HIV-1 RNA NAT testing. According to the results of the HIV seroconversion panels, the Elecsys HIV Combi PT could detect seroconversion at the same bleed or at least one bleed earlier compared to the other two assays. Among the 675 clinical samples, most results were consistent except for one specimen with a false-negative result using Elecsys HIV Combi assay. In conclusion, the Elecsys HIV Combi PT has shown satisfactory sensitivity and specificity to be a screening test for HIV infection.

  7. Clinical Evaluation of Shilajatu Rasayana in patients with HIV Infection.

    Science.gov (United States)

    Gupta, G D; Sujatha, N; Dhanik, Ajay; Rai, N P

    2010-01-01

    AIDS is one of the serious global health concerns caused by Human Immuno Deficiency(HIV) virus and is predominantly a sexually transmitted disease. Currently there is no vaccine or cure for AIDS still Anti Retroviral Therapy (ART) is successful. It reduces both the mortality and the morbidity of HIV infection, but is expensive and inaccessible in many countries. However intense the therapy may be, HIV virus is rarely eliminated, and drug resistance is a major setback during long-term therapy. The development of new drugs and strategies and exploring alternative systems of medicine for antiviral herbs or drugs is the need of the age to improve treatment outcomes. Ayurveda describes many diseases which incorporate HIV like illness e.g. Rajayakshma, Ojo Kshaya, Sannipata jwara etc. HIV infection affects multisystems, chiefly the Immune System which can be correlated to Ojo Kshaya. Rasayana Chikitsa is the frontline therapy employed to treat Ojus disorders. Therefore Shilajatu (Mineral pitch), Centella asiatica (Mandukaparni), Tinospora cordifolia (Guduchi) and Emblica officinalis (Amalaki), well known for their Immuno-modulator and antioxidant properties were selected to evaluate their role on immune system. The study was carried on 20 patients from OPD and IPD of Kayachikitsa, S.S.Hospital, IMS, BHU and was randomly allocated into Treated group (Shilajatu+ART) and Control group (ART). Treated Group responded better to ART both clinically and biochemically. The results show that Shilajatu decreases the recurrent resistance of HIV virus to ART and improves the outcome of the therapy.

  8. Evaluation and Diagnosis of HIV-Associated Lung Disease.

    Science.gov (United States)

    Maximous, Stephanie; Huang, Laurence; Morris, Alison

    2016-04-01

    There are myriad pulmonary conditions associated with HIV, ranging from acute infections to chronic noncommunicable diseases. The epidemiology of these diseases has changed significantly in the era of widespread antiretroviral therapy. Evaluation of the HIV-infected patient involves assessment of the severity of illness and a thorough yet efficient pursuit of definitive diagnosis, which may involve multiple etiologies simultaneously. Important clues to a diagnosis include medical and social history, demographic details such as travel and geography of residence, substance use, sexual practices, and domiciliary and incarceration status. CD4 cell count is a tremendously useful measure of immune function and risk for HIV-related diseases, and helps narrow down the differential. Careful history of current symptoms and physical examination with particular attention to extrapulmonary signs are crucial early steps. Many adjunctive laboratory studies can suggest or rule out particular diagnoses. Pulmonary function testing (PFT) may aid in characterization of several chronic noninfectious illnesses accelerated by HIV. Chest radiograph and computed tomography (CT) scan allow for classification of diseases by pathognomonic imaging patterns, although many infectious conditions present atypically, particularly with lower CD4 counts. Ultimately, definitive diagnosis with sputum, bronchoscopy with bronchoalveolar lavage, or lung tissue is often needed. It is of utmost importance to maintain a high degree of suspicion for HIV in otherwise undiagnosed patients, as the first presentation of HIV may be via an acute pulmonary illness. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  9. Physiological Mg(2+) Conditions Significantly Alter the Inhibition of HIV-1 and HIV-2 Reverse Transcriptases by Nucleoside and Non-Nucleoside Inhibitors in Vitro.

    Science.gov (United States)

    Achuthan, Vasudevan; Singh, Kamlendra; DeStefano, Jeffrey J

    2017-01-10

    Reverse transcriptases (RTs) are typically assayed in vitro with 5-10 mM Mg(2+), whereas the free Mg(2+) concentration in cells is much lower. Artificially high Mg(2+) concentrations used in vitro can misrepresent different properties of human immunodeficiency virus (HIV) RT, including fidelity, catalysis, pausing, and RNase H activity. Here, we analyzed nucleoside (NRTIs) and non-nucleoside RT inhibitors (NNRTIs) in primer extension assays at different concentrations of free Mg(2+). At low concentrations of Mg(2+), NRTIs and dideoxynucleotides (AZTTP, ddCTP, ddGTP, and 3TCTP) inhibited HIV-1 and HIV-2 RT synthesis less efficiently than they did with large amounts of Mg(2+), whereas inhibition by the "translocation-defective RT inhibitor" EFdA (4'-ethynyl-2-fluoro-2'-deoxyadenosine) was unaffected by Mg(2+) concentrations. Steady-state kinetic analyses revealed that the reduced level of inhibition at low Mg(2+) concentrations resulted from a 3-9-fold (depending on the particular nucleotide and inhibitor) less efficient incorporation (based on kcat/Km) of these NRTIs under this condition compared to incorporation of natural dNTPs. In contrast, EFdATP was incorporated with an efficiency similar to that of its analogue dATP at low Mg(2+) concentrations. Unlike NRTIs, NNRTIs (nevirapine, efavirenz, and rilviripine), were approximately 4-fold (based on IC50 values) more effective at low than at high Mg(2+) concentrations. Drug-resistant HIV-1 RT mutants also displayed the Mg(2+)-dependent difference in susceptibility to NRTIs and NNRTIs. In summary, analyzing the efficiency of inhibitors under more physiologically relevant low-Mg(2+) conditions yielded results dramatically different from those from measurements using commonly employed high-Mg(2+) in vitro conditions. These results also emphasize differences in Mg(2+) sensitivity between the translocation inhibitor EFdATP and other NRTIs.

  10. Specific reverse transcriptase slippage at the HIV ribosomal frameshift sequence: potential implications for modulation of GagPol synthesis.

    Science.gov (United States)

    Penno, Christophe; Kumari, Romika; Baranov, Pavel V; van Sinderen, Douwe; Atkins, John F

    2017-09-29

    Synthesis of HIV GagPol involves a proportion of ribosomes translating a U6A shift site at the distal end of the gag gene performing a programmed -1 ribosomal frameshift event to enter the overlapping pol gene. In vitro studies here show that at the same shift motif HIV reverse transcriptase generates -1 and +1 indels with their ratio being sensitive to the relative concentration ratio of dNTPs specified by the RNA template slippage-prone sequence and its 5' adjacent base. The GGG sequence 3' adjacent to the U6A shift/slippage site, which is important for ribosomal frameshifting, is shown here to limit reverse transcriptase base substitution and indel 'errors' in the run of A's in the product. The indels characterized here have either 1 more or less A, than the corresponding number of template U's. cDNA with 5 A's may yield novel Gag product(s), while cDNA with an extra base, 7 A's, may only be a minor contributor to GagPol polyprotein. Synthesis of a proportion of non-ribosomal frameshift derived GagPol would be relevant in efforts to identify therapeutically useful compounds that perturb the ratio of GagPol to Gag, and pertinent to the extent in which specific polymerase slippage is utilized in gene expression. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  11. Variability in non-nucleoside reverse transcriptase and protease inhibitor concentrations among HIV-infected adults in routine clinical practice

    Science.gov (United States)

    Moltó, José; Blanco, Asunción; Miranda, Cristina; Miranda, José; Puig, Jordi; Valle, Marta; DelaVarga, Meritxell; Fumaz, Carmina R; Barbanoj, Manuel José; Clotet, Bonaventura

    2006-01-01

    What is already known about this subject The concentration of protease and non-nucleoside reverse transcriptase inhibtors in plasma has been related to both efficacy and toxicity. Most antiretroviral concentration data come from selected populations of patients undergoing therapeutic drug monitoring programmes, which may overestimate interindividual variability. What this study adds Our study has demonstrated the large interindividual variability in antiretroviral drug concentrations in an unselected population of patients during routine clinical practice. These results may provide interesting information to clinicians for the management of antiretroviral therapy in HIV-infected patients. Aims The objective of this study was to assess interindividual variability in trough concentrations of plasma of non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI) among HIV-infected adults in a routine outpatient setting. Methods One hundred and seventeen patients who attended our clinic for routine blood tests, and who were receiving antiretroviral therapy which included NNRTI or PI were studied. Patients were not informed that drug concentrations were going to be measured until blood sampling. The times of the last antiretroviral dose and of blood sampling were recorded. Drug concentrations were considered optimal if they were above the proposed minimum effective value. In addition, efavirenz, nevirapine and atazanavir concentrations were considered potentially toxic if they were >4.0 mg l−1, >6.0 mg l−1 and >0.85 mg l−1, respectively. Results Overall, interindividual variability of NNRTI and PI concentrations in plasma was approximately 50%, and only 68.4% of the patients had drug concentrations within the proposed therapeutic range. Poor adherence explained only 35% of subtherapeutic drug concentrations. Conclusion Interindividual variability in trough concentrations of NNRTI and PI among HIV-infected adults is large in routine clinical

  12. Inhibition of both HIV-1 reverse transcription and gene expression by a cyclic peptide that binds the Tat-transactivating response element (TAR RNA.

    Directory of Open Access Journals (Sweden)

    Matthew S Lalonde

    2011-05-01

    Full Text Available The RNA response element TAR plays a critical role in HIV replication by providing a binding site for the recruitment of the viral transactivator protein Tat. Using a structure-guided approach, we have developed a series of conformationally-constrained cyclic peptides that act as structural mimics of the Tat RNA binding region and block Tat-TAR interactions at nanomolar concentrations in vitro. Here we show that these compounds block Tat-dependent transcription in cell-free systems and in cell-based reporter assays. The compounds are also cell permeable, have low toxicity, and inhibit replication of diverse HIV-1 strains, including both CXCR4-tropic and CCR5-tropic primary HIV-1 isolates of the divergent subtypes A, B, C, D and CRF01_AE. In human peripheral blood mononuclear cells, the cyclic peptidomimetic L50 exhibited an IC(50 ∼250 nM. Surprisingly, inhibition of LTR-driven HIV-1 transcription could not account for the full antiviral activity. Timed drug-addition experiments revealed that L-50 has a bi-phasic inhibition curve with the first phase occurring after HIV-1 entry into the host cell and during the initiation of HIV-1 reverse transcription. The second phase coincides with inhibition of HIV-1 transcription. Reconstituted reverse transcription assays confirm that HIV-1 (- strand strong stop DNA synthesis is blocked by L50-TAR RNA interactions in-vitro. These findings are consistent with genetic evidence that TAR plays critical roles both during reverse transcription and during HIV gene expression. Our results suggest that antiviral drugs targeting TAR RNA might be highly effective due to a dual inhibitory mechanism.

  13. Performance evaluation of reverse logistics: A case of LPG agency

    Directory of Open Access Journals (Sweden)

    Kottala Sri Yogi

    2015-12-01

    Full Text Available Majority of the manufacturing companies are incorporating the practice of reverse logistics in their value chain. In manufacturing processes, the concept of reverse logistics plays a vital role in enhancing the company’s profit margin for sustainable business growth. For every company, there is a need of performance measurement system to be established as successful business tool. However to predict better results, how smartly the inputs for the transformation or business process are being effectively and efficiently used has to be analyzed. Drawing attention to the growing popularity in adapting the best practices of reverse logistics among different manufacturing industries, this paper aims to build a methodology in order to measure the performance of liquefied petroleum gas (LPG agencies. To do so, it has undertaken the Asian emerging market-India as a case study and improved the understanding of performance measurement in reverse logistics to refilling LPG cylinders. Further, it has suggested a framework affecting the implementation of reverse logistics activities and its network.

  14. 8-Modified-2'-deoxyadenosine analogues induce delayed polymerization arrest during HIV-1 reverse transcription.

    Directory of Open Access Journals (Sweden)

    Valérie Vivet-Boudou

    Full Text Available The occurrence of resistant viruses to any of the anti-HIV-1 compounds used in the current therapies against AIDS underlies the urge for the development of new drug targets and/or new drugs acting through novel mechanisms. While all anti-HIV-1 nucleoside analogues in clinical use and in clinical trials rely on ribose modifications for activity, we designed nucleosides with a natural deoxyribose moiety and modifications of position 8 of the adenine base. Such modifications might induce a steric clash with helix αH in the thumb domain of the p66 subunit of HIV-1 RT at a distance from the catalytic site, causing delayed chain termination. Eleven new 2'-deoxyadenosine analogues modified on position 8 of the purine base were synthesized and tested in vitro and in cell-based assays. In this paper we demonstrate for the first time that chemical modifications on position 8 of 2'-deoxyadenosine induce delayed chain termination in vitro, and also inhibit DNA synthesis when incorporated in a DNA template strand. Furthermore, one of them had moderate anti-HIV-1 activity in cell-culture. Our results constitute a proof of concept indicating that modification on the base moiety of nucleosides can induce delayed polymerization arrest and inhibit HIV-1 replication.

  15. Bilateral Visual Loss as Presenting Symptom of Posterior Reversible Encephalopathy Syndrome in a Patient with HIV/Tuberculosis Coinfection: A Case Report

    Directory of Open Access Journals (Sweden)

    S. Guerriero

    2012-01-01

    Full Text Available Posterior reversible encephalopathy syndrome (PRES is a neurotoxic state accompanied by a unique brain imaging pattern. This cliniconeuroradiological entity usually presents with visual disturbances (cortical blindness, homonymous hemianopia, visual neglect, and blurred vision along with neurotoxic manifestations. Only a few cases of PRES have previously been reported in patients with advanced HIV disease. The authors describe a case of posterior reversible encephalopathy syndrome (PRES in a patient with advanced HIV/TBC infection who developed a neurotoxic state following TB and ART therapy initiation. They present a comprehensive review of the literature and discuss the pathogenetic hypotheses.

  16. Evaluation of School- and Community-Based HIV Prevention ...

    African Journals Online (AJOL)

    The Ministry of Education approved Family Life and HIV Education (FLHE) programme delivered in Junior Secondary Schools and a community-based initiative to raise AIDS Competency of rural communities were evaluated using a clustered randomized control trial and mixed qualitative-quantitative methods. Ten schools ...

  17. Evaluation of liver function tests of HIV positive patients on ...

    African Journals Online (AJOL)

    Liver enzymes-alanine and aspartate aminotransferases and alkaline phosphatase (AST, ALT and ALP), bilirubin and serum proteins were determined using standard laboratory methods and these parameters were used to evaluate the liver function of human immunodeficiency virus (HIV)- positive patients receiving ...

  18. High-Resolution Structures of HIV-1 Reverse Transcriptase/TMC278 Complexes: Strategic Flexibility Explains Potency Against Resistance Mutations

    Energy Technology Data Exchange (ETDEWEB)

    Das,K.; Bauman, J.; Clark, Jr., A.; Frenkel, Y.; Lewi, P.; Shatkin, A.; Hughes, S.; Arnold, E.

    2008-01-01

    TMC278 is a diarylpyrimidine (DAPY) nonnucleoside reverse transcriptase inhibitor (NNRTI) that is highly effective in treating wild-type and drug-resistant HIV-1 infections in clinical trials at relatively low doses ({approx}25-75 mg/day). We have determined the structure of wild-type HIV-1 RT complexed with TMC278 at 1.8 Angstroms resolution, using an RT crystal form engineered by systematic RT mutagenesis. This high-resolution structure reveals that the cyanovinyl group of TMC278 is positioned in a hydrophobic tunnel connecting the NNRTI-binding pocket to the nucleic acid-binding cleft. The crystal structures of TMC278 in complexes with the double mutant K103N/Y181C (2.1 Angstroms ) and L100I/K103N HIV-1 RTs (2.9 Angstroms ) demonstrated that TMC278 adapts to bind mutant RTs. In the K103N/Y181C RT/TMC278 structure, loss of the aromatic ring interaction caused by the Y181C mutation is counterbalanced by interactions between the cyanovinyl group of TMC278 and the aromatic side chain of Y183, which is facilitated by an {approx}1.5 Angstroms shift of the conserved Y183MDD motif. In the L100I/K103N RT/TMC278 structure, the binding mode of TMC278 is significantly altered so that the drug conforms to changes in the binding pocket primarily caused by the L100I mutation. The flexible binding pocket acts as a molecular 'shrink wrap' that makes a shape complementary to the optimized TMC278 in wild-type and drug-resistant forms of HIV-1 RT. The crystal structures provide a better understanding of how the flexibility of an inhibitor can compensate for drug-resistance mutations.

  19. In search of a treatment for HIV--current therapies and the role of non-nucleoside reverse transcriptase inhibitors (NNRTIs).

    Science.gov (United States)

    Reynolds, Chevonne; de Koning, Charles B; Pelly, Stephen C; van Otterlo, Willem A L; Bode, Moira L

    2012-07-07

    The human immunodeficiency virus (HIV) causes AIDS (acquired immune deficiency syndrome), a disease in which the immune system progressively deteriorates, making sufferers vulnerable to all manner of opportunistic infections. Currently, world-wide there are estimated to be 34 million people living with HIV, with the vast majority of these living in sub-Saharan Africa. Therefore, an important research focus is development of new drugs that can be used in the treatment of HIV/AIDS. This review gives an overview of the disease and addresses the drugs currently used for treatment, with specific emphasis on new developments within the class of allosteric non-nucleoside reverse transcriptase inhibitors (NNRTIs).

  20. Mutations in HIV-1 Reverse Transcriptase Affect the Errors Made in a Single Cycle of Viral Replication

    Science.gov (United States)

    Abram, Michael E.; Ferris, Andrea L.; Das, Kalyan; Quinoñes, Octavio; Shao, Wei; Tuske, Steven; Alvord, W. Gregory; Arnold, Eddy

    2014-01-01

    ABSTRACT The genetic variation in HIV-1 in patients is due to the high rate of viral replication, the high viral load, and the errors made during viral replication. Some of the mutations in reverse transcriptase (RT) that alter the deoxynucleoside triphosphate (dNTP)-binding pocket, including those that confer resistance to nucleoside/nucleotide analogs, affect dNTP selection during replication. The effects of mutations in RT on the spectrum (nature, position, and frequency) of errors made in vivo are poorly understood. We previously determined the mutation rate and the frequency of different types of mutations and identified hot spots for mutations in a lacZα (the α complementing region of lacZ) reporter gene carried by an HIV-1 vector that replicates using wild-type RT. We show here that four mutations (Y115F, M184V, M184I, and Q151M) in the dNTP-binding pocket of RT that had relatively small effects on the overall HIV-1 mutation rate (less than 3-fold compared to the wild type) significantly increased mutations at some specific positions in the lacZα reporter gene. We also show that changes in a sequence that flanks the reporter gene can affect the mutations that arise in the reporter. These data show that changes either in HIV-1 RT or in the sequence of the nucleic acid template can affect the spectrum of mutations made during viral replication. This could, by implication, affect the generation of drug-resistant mutants and immunological-escape mutants in patients. IMPORTANCE RT is the viral enzyme that converts the RNA genome of HIV into DNA. Errors made during replication allow the virus to escape from the host's immune system and to develop resistance to the available anti-HIV drugs. We show that four different mutations in RT which are known to be associated with resistance to anti-RT drugs modestly increased the overall frequency of errors made during viral replication. However, the increased errors were not uniformly distributed; the additional errors

  1. Evaluation of an alternative supplemental testing strategy for HIV diagnosis by retrospective analysis of clinical HIV testing data.

    Science.gov (United States)

    Styer, Linda M; Sullivan, Timothy J; Parker, Monica M

    2011-12-01

    Because newer screening assays are more sensitive than traditional confirmatory assays, a new HIV testing algorithm was proposed by the Centers for Disease Control and Prevention (CDC) and Association of Public Health Laboratories (APHL) in 2010 to replace the current enzyme immunoassay (EIA)-Western blot (WB) algorithm that was established in 1989. The new algorithm includes a sensitive screening EIA and a HIV-1/HIV-2 differentiation immunoassay (Multispot) for confirmation. Concordant reactive specimens are reported as HIV-1 and/or HIV-2 positive; those with discordant results receive nucleic acid testing (NAT). Our laboratory uses all components of both the current and proposed algorithms and conducted a retrospective analysis of test results to compare algorithm performance. All available test results from 38,257 specimens were analyzed, of which 36,598 were EIA non-reactive. Of 1659 EIA-reactive specimens, 1578 were defined by our laboratory as HIV-1 positive, 5 as HIV-2 positive, 69 as negative and 7 as unknown. These results were used to evaluate both algorithms. Under the proposed algorithm, all 1578 HIV-1-positive specimens would be reported as 'HIV-1 positive', whereas the current algorithm only confirmed 98% (1546/1578). The proposed algorithm produced fewer inconclusive results (9 vs 48); consequently, fewer follow-up specimens would be needed. The current algorithm produced one false-positive result whereas the proposed algorithm produced four. Two of these four false-positive results could be eliminated by requiring NAT for specimens with discordant HIV-1 results on the Multispot. Although both algorithms identified all HIV-2-positive specimens, the current algorithm required an additional 112 tests. The CDC/APHL HIV testing algorithm, proposed in 2010, outperformed the current algorithm because it was more sensitive for detecting HIV-1 infection, provided a greater number of definitive HIV results, and detected HIV-2 more efficiently. Copyright

  2. Synthesis of Novel Uracil Non-Nucleoside Derivatives as Potential Reverse Transcriptase Inhibitors of HIV-1

    DEFF Research Database (Denmark)

    El-Brollosy, Nasser R.; Al-Deeb, Omar. A.; El-Emam, Ali A.

    2009-01-01

    with cyclopropylmethyloxymethyl 9a-d, 2-phenylethyloxymethyl 9e-h, and 3-phenylprop-1-yloxymethyl 9i-l were prepared on treatment of the corresponding uracils with the appropriate acetals 8a-c. Some of the tested compounds showed good activity against HIV-1 wild type. Among them, 1-cyclopropylmethyloxymethyl-5-ethyl-6......-(3,5-dimethylbenzyl)uracil 9c and 5-ethyl-6-(3,5-dimethylbenzyl)-1-(2-phenylethyloxymethyl)uracil 9g showed inhibitory potency equally to emivirine against HIV-1 wild type. Furthermore, compounds 9c and 9g showed marginal better activity against NNRTI resistant mutants than emivirine....

  3. Intraoperative Evaluation of Reverse Bypass Using a Naturally Formed "Bonnet" Superficial Temporal Artery: Technical Note.

    Science.gov (United States)

    Nagm, Alhusain; Horiuchi, Tetsuyoshi; Hasegawa, Takatoshi; Hongo, Kazuhiro

    2016-04-01

    In reverse bypass that used a naturally formed "bonnet" superficial temporal artery, intraoperative volume flow measurement quantifies flow augmentation after revascularization, confirms flow preservation, and identifies inadvertent vessel compromise. A 75-year-old man presented with transient ischemic attacks attributed to right internal carotid artery stenosis. He underwent successful reverse bypass via a naturally formed "bonnet" superficial temporal artery middle cerebral artery bypass. As the result of proper intraoperative volume flow evaluation, a successful reverse bypass was achieved. Modification of the intraoperative stroke risk and prediction of the long-term patency after reverse bypass can be achieved by meticulous intraoperative blood flow evaluation. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Site-directed mutagenesis of the conserved Asp-443 and Asp-498 carboxy-terminal residues of HIV-1 reverse transcriptase.

    OpenAIRE

    Mizrahi, V; Usdin, M T; Harington, A; Dudding, L R

    1990-01-01

    Substitution of the conserved Asp-443 residue of HIV-1 reverse transcriptase by asparagine specifically suppressed the ribonuclease H activity of the enzyme without affecting the reverse transcriptase activity, suggesting involvement of this ionizable residue at the ribonuclease H active site. An analogous asparagine substitution of the Asp-498 residue yielded an unstable enzyme that was difficult to enzymatically characterize. However, the instability caused by the Asn-498 mutation was relie...

  5. Drug resistance-related mutations T369V/I in the connection subdomain of HIV-1 reverse transcriptase severely impair viral fitness.

    Science.gov (United States)

    Wang, Zheng; Zhang, Junli; Li, Fan; Ji, Xiaolin; Liao, Lingjie; Ma, Liying; Xing, Hui; Feng, Yi; Li, Dan; Shao, Yiming

    2017-04-02

    Fitness is a key parameter in the measurement of transmission capacity of individual drug-resistant HIV. Drug-resistance related mutations (DRMs) T369V/I and A371V in the connection subdomain (CN) of reverse transcriptase (RT) occur at higher frequencies in the individuals experiencing antiretroviral therapy failure. Here, we evaluated the effects of T369V/I and A371V on viral fitness, in the presence or in the absence of thymidine analogue resistance-associated mutations (TAMs) and assessed the effect of potential RT structure-related mechanism on change in viral fitness. Mutations T369V/I, A371V, alone or in combination with TAMs were introduced into a modified HIV-1 infectious clone AT1 by site-directed mutagenesis. Then, experiments on mutant and wild-type virus AT2 were performed separately using a growth-competition assay, and then the relative fitness was calculated. Structural analysis of RT was conducted using Pymol software. Results showed that T369V/I severely impaired the relative virus fitness, and A371V compensated for the viral fitness reduction caused by TAMs. Structural modeling of RT suggests that T369V/I substitutions disrupt powerful hydrogen bonds formed by T369 and V365 in p51 and p66. This study indicates that the secondary DRMs within CN might efficiently damage viral fitness, and provides valuable information for clinical surveillance and prevention of HIV-1 strains carrying these DRMs. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Long-term survival with HIV-infection : the reverse side of the medal

    NARCIS (Netherlands)

    Jong, E.

    2010-01-01

    The introduction of highly effective combined antiretroviral therapy (cART) in 1996 resulted in effective, long-term suppression of HIV with consequent recovery of the patient’s immune system. Although the enthusiasm about the effectiveness of current cART remains strong, the ongoing work in the

  7. Structure-based virtual screening efforts against HIV-1 reverse transcriptase to introduce the new potent non-nucleoside reverse transcriptase inhibitor

    Science.gov (United States)

    Hosseini, Yaser; Mollica, Adriano; Mirzaie, Sako

    2016-12-01

    The human immunodeficiency virus (HIV) which is strictly related to the development of AIDS, is treated by a cocktail of drugs, but due its high propensity gain drug resistance, the rational development of new medicine is highly desired. Among the different mechanism of action we selected the reverse transcriptase (RT) inhibition, for our studies. With the aim to identify new chemical entities to be used for further rational drug design, a set of 3000 molecules from the Zinc Database have been screened by docking experiments using AutoDock Vina software. The best ranked compounds with respect of the crystallographic inhibitor MK-4965 resulted to be five compounds, and the best among them was further tested by molecular dynamics (MD) simulation. Our results indicate that comp1 has a stronger interaction with the subsite p66 of RT than MK-4965 and that both are able to stabilize specific conformational changes of the RT 3D structure, which may explain their activity as inhibitors. Therefore comp1 could be a good candidate for biological tests and further development.

  8. Developing weighted criteria to evaluate lean reverse logistics through analytical network process

    Science.gov (United States)

    Zagloel, Teuku Yuri M.; Hakim, Inaki Maulida; Krisnawardhani, Rike Adyartie

    2017-11-01

    Reverse logistics is a part of supply chain that bring materials from consumers back to manufacturer in order to gain added value or do a proper disposal. Nowadays, most companies are still facing several problems on reverse logistics implementation which leads to high waste along reverse logistics processes. In order to overcome this problem, Madsen [Framework for Reverse Lean Logistics to Enable Green Manufacturing, Eco Design 2009: 6th International Symposium on Environmentally Conscious Design and Inverse Manufacturing, Sapporo, 2009] has developed a lean reverse logistics framework as a step to eliminate waste by implementing lean on reverse logistics. However, the resulted framework sets aside criteria used to evaluate its performance. This research aims to determine weighted criteria that can be used as a base on reverse logistics evaluation by considering lean principles. The resulted criteria will ensure reverse logistics are kept off from waste, thus implemented efficiently. Analytical Network Process (ANP) is used in this research to determine the weighted criteria. The result shows that criteria used for evaluation lean reverse logistics are Innovation and Learning (35%), Economic (30%), Process Flow Management (14%), Customer Relationship Management (13%), Environment (6%), and Social (2%).

  9. Evaluation of Salivary Vitamin C and Catalase in HIV Positive and Healthy HIV Negative Control Group.

    Science.gov (United States)

    Ahmadi-Motamayel, Fatemeh; Vaziri-Amjad, Samaneh; Goodarzi, Mohammad Taghi; Poorolajal, Jalal

    2017-01-01

    Saliva is a complex oral biologic fluid secreted by major and minor salivary glands. Saliva has immunological, enzymatic and antioxidant defense mechanisms. Infection with human immunodeficiency virus (HIV) is a life-threatening disease. The aim of this study was to evaluate salivary vitamin C and catalase levels in HIV-positive patients in comparison to a healthy control group. Forty-nine HIV-infected individuals and 49 healthy subjects were selected. Five mL of unstimulated saliva was collected in 5 minutes using a sterilized Falcon tube with Navazesh method. Catalase and vitamin C levels were assessed by spectrophotometric assay. Data were analyzed with STATA 12. Salivary catalase levels were 7.99±2.40 and 8.37±1.81 in the case and control groups, respectively. Catalase level was lower in the case group but the difference was not statistically significant (P=0.380). Salivary vitamin C levels in the case and control groups were 3.76±1.92 and 4.87±2.20, respectively (P=0.009). HIV can alter salivary antioxidant capacity as well as vitamin C and catalase levels. Saliva may reflect serum antioxidative changes in these patients. Therefore, further research is necessary on salivary and serum oxidants and the antioxidant changes. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  10. Coccinin, an antifungal peptide with antiproliferative and HIV-1 reverse transcriptase inhibitory activities from large scarlet runner beans.

    Science.gov (United States)

    Ngai, Patrick H K; Ng, T B

    2004-12-01

    An antifungal peptide, designated coccinin, with a molecular mass of 7kDa and an N-terminal sequence resembling those of defensins, was purified from the seeds of large scarlet runner beans (Phaseolus coccineus cv. 'Major'). The peptide isolated was unadsorbed on DEAE-cellulose, and adsorbed on Affi-gel blue gel and Mono S. The peptide excerted antifungal activity on a number of fungal species including Botrytis cinerea, Coprinus comatus, Fusarium oxysporum, Mycosphaerella arachidicola, Physalospora piricola, and Rhizoctonia solani. It also inhibited proliferation in the leukemia cell lines HL60 and L1210, and reduced the activity of HIV-1 reverse transcriptase. However, it did not affect proliferation of mouse splenocytes.

  11. Phaseococcin, an antifungal protein with antiproliferative and anti-HIV-1 reverse transcriptase activities from small scarlet runner beans.

    Science.gov (United States)

    Ngai, Patrick H K; Ng, T B

    2005-04-01

    From the seeds of small scarlet runner beans (Phaseolus coccineus 'Minor'), an antifungal protein with an N-terminal sequence homologous to those of defensins was isolated. The antifungal protein bound to Affi-gel blue gel and Mono S but it did not bind to DEAE-cellulose. It was further purified by gel filtration on a Superdex peptide column. It exhibited a molecular mass of 5422 Da as determined by mass spectrometry. The protein, designated as phaseococcin, suppressed mycelial growth in a number of fungi including Botrytis cinerea, Coprinus comatus, Fusarium oxysporum, Mycosphaerella arachidicola, Physalospora piricola, and Rhizoctonia solani. It also inhibited proliferation in several Bacillus species and the leukemia cell lines HL60 and L1210 and curtailed the activity of HIV-1 reverse transcriptase. It did not affect proliferation of mouse splenocytes and neither did it inhibit protein synthesis in a cell-free rabbit reticulocyte lysate system.

  12. Interpatient variability in the pharmacokinetics of the HIV non-nucleoside reverse transcriptase inhibitor efavirenz: the effect of gender, race, and CYP2B6 polymorphism

    NARCIS (Netherlands)

    Burger, D.M.; Heiden, I. van der; Porte, C.J.L. la; Ende, M. van der; Groeneveld, P.; Richter, C.; Koopmans, P.; Kroon, F.; Sprenger, H.; Lindemans, J.; Schenk, P.; Schaik, R. van

    2006-01-01

    AIMS: To characterize the demographic and pharmacogenetic factors that influence interpatient variability in the plasma concentrations of the HIV non-nucleoside reverse transcriptase inhibitor efavirenz. METHODS: Data from all samples analyzed for efavirenz in our TDM service in 2002 and 2003 were

  13. Interpatient variability in the pharmacokinetics of the HIV non-nucleoside reverse transcriptase inhibitor efavirenz : the effect of gender, race, and CYP2B6 polymorphism

    NARCIS (Netherlands)

    Burger, D; van der Heiden, [No Value; la Porte, C; van der Ende, Marchina E.; Groeneveld, P; Richter, C; Koopmans, P; Sprenger, H; Lindemans, J; Schenk, P; van Schaik, R

    Aims To characterize the demographic and pharmacogenetic factors that influence interpatient variability in the plasma concentrations of the HIV non-nucleoside reverse transcriptase inhibitor efavirenz. Methods Data from all samples analyzed for efavirenz in our TDM service in 2002 and 2003 were

  14. Novel (2,6-difluorophenyl)(2-(phenylamino)pyrimidin-4-yl) methanones with restricted conformation as potent non-nucleoside reverse transcriptase inhibitors against HIV-1

    Czech Academy of Sciences Publication Activity Database

    Šimon, Petr; Baszczyňski, Ondřej; Šaman, David; Stepan, G.; Hu, E.; Lansdon, E. B.; Jansa, P.; Janeba, Zlatko

    2016-01-01

    Roč. 122, Oct 21 (2016), s. 185-195 ISSN 0223-5234 Institutional support: RVO:61388963 Keywords : diarylpyrimidine (DAPY) * etravirine * human immunodeficiency virus ( HIV ) * non-nucleoside reverse transcriptase inhibitors * NNRTIs * rilpivirine Subject RIV: CC - Organic Chemistry Impact factor: 4.519, year: 2016

  15. Measuring enzymatic HIV-1 susceptibility to two reverse transcriptase inhibitors as a rapid and simple approach to HIV-1 drug-resistance testing.

    Directory of Open Access Journals (Sweden)

    Dieter Hoffmann

    Full Text Available Simple and cost-effective approaches for HIV drug-resistance testing are highly desirable for managing increasingly expanding HIV-1 infected populations who initiate antiretroviral therapy (ART, particularly in resource-limited settings. Non-nucleoside reverse trancriptase inhibitor (NNRTI-based regimens with an NRTI backbone containing lamivudine (3TC or emtricitabine (FTC are preferred first ART regimens. Failure with these drug combinations typically involves the selection of NNRTI- and/or 3TC/FTC-resistant viruses. Therefore, the availability of simple assays to measure both types of drug resistance is critical. We have developed a high throughput screening test for assessing enzymatic resistance of the HIV-1 RT in plasma to 3TC/FTC and NNRTIs. The test uses the sensitive "Amp-RT" assay with a newly-developed real-time PCR format to screen biochemically for drug resistance in single reactions containing either 3TC-triphosphate (3TC-TP or nevirapine (NVP. Assay cut-offs were defined based on testing a large panel of subtype B and non-subtype B clinical samples with known genotypic profiles. Enzymatic 3TC resistance correlated well with the presence of M184I/V, and reduced NVP susceptibility was strongly associated with the presence of K103N, Y181C/I, Y188L, and G190A/Q. The sensitivity and specificity for detecting resistance were 97.0% and 96.0% in samples with M184V, and 97.4% and 96.2% for samples with NNRTI mutations, respectively. We further demonstrate the utility of an HIV capture method in plasma by using magnetic beads coated with CD44 antibody that eliminates the need for ultracentifugation. Thus our results support the use of this simple approach for distinguishing WT from NNRTI- or 3TC/FTC-resistant viruses in clinical samples. This enzymatic testing is subtype-independent and can assist in the clinical management of diverse populations particularly in resource-limited settings.

  16. Computational Analysis of Molecular Interaction Networks Underlying Change of HIV-1 Resistance to Selected Reverse Transcriptase Inhibitors.

    Science.gov (United States)

    Kierczak, Marcin; Dramiński, Michał; Koronacki, Jacek; Komorowski, Jan

    2010-12-12

    Despite more than two decades of research, HIV resistance to drugs remains a serious obstacle in developing efficient AIDS treatments. Several computational methods have been developed to predict resistance level from the sequence of viral proteins such as reverse transcriptase (RT) or protease. These methods, while powerful and accurate, give very little insight into the molecular interactions that underly acquisition of drug resistance/hypersusceptibility. Here, we attempt at filling this gap by using our Monte Carlo feature selection and interdependency discovery method (MCFS-ID) to elucidate molecular interaction networks that characterize viral strains with altered drug resistance levels. We analyzed a number of HIV-1 RT sequences annotated with drug resistance level using the MCFS-ID method. This let us expound interdependency networks that characterize change of drug resistance to six selected RT inhibitors: Abacavir, Lamivudine, Stavudine, Zidovudine, Tenofovir and Nevirapine. The networks consider interdependencies at the level of physicochemical properties of mutating amino acids, eg,: polarity. We mapped each network on the 3D structure of RT in attempt to understand the molecular meaning of interacting pairs. The discovered interactions describe several known drug resistance mechanisms and, importantly, some previously unidentified ones. Our approach can be easily applied to a whole range of problems from the domain of protein engineering. A portable Java implementation of our MCFS-ID method is freely available for academic users and can be obtained at: http://www.ipipan.eu/staff/m.draminski/software.htm.

  17. Activation of the human nuclear xenobiotic receptor PXR by the reverse transcriptase-targeted anti-HIV drug PNU-142721

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Yuan; Redinbo, Matthew R. (UNC)

    2012-10-09

    The human pregnane X receptor (PXR) is a member of the nuclear receptor superfamily of ligand-regulated transcription factors. PXR responds to a structurally diverse variety of endogenous and xenobiotic compounds, and coordinates the expression of genes central to the metabolism and excretion of potentially harmful chemicals, including human therapeutics. The reverse transcriptase inhibitor PNU-142721 has been designed to treat human immunodeficiency virus (HIV) infection. Although this compound has anti-HIV activity, it was established using cell-based assays that PNU-142721 is an efficacious PXR agonist. We present here the 2.8 {angstrom} resolution crystal structure of the human PXR ligand-binding domain in complex with PNU-142721. PXR employs one hydrogen bond and fourteen van der Waals contacts to interact with the ligand, but allows two loops adjacent to the ligand-binding pocket to remain disordered in the structure. These observations highlight the role structural flexibility plays in PXR's promiscuous responses to xenobiotics. The crystal structure also explains why PNU-173575, a thiomethyl metabolite of PNU-142721, exhibits enhanced PXR activation relative to the unmodified compound and why PNU-142721 can also activate rat PXR. Taken together, the results presented here elucidate the structural basis for PXR activation by PNU-142721 and related chemicals.

  18. Variability in non-nucleoside reverse transcriptase and protease inhibitors concentrations among HIV-infected adults in routine clinical practice

    Science.gov (United States)

    Moltó, José; Blanco, Asunción; Miranda, Cristina; Miranda, José; Puig, Jordi; Valle, Marta; DelaVarga, Meritxell; Fumaz, Carmina R; Barbanoj, Manuel José; Clotet, Bonaventura

    2007-01-01

    Aims The objective of this study was to assess interindividual variability in plasma trough concentrations of non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI) among HIV-infected adults in an outpatient routine clinical practice setting. Methods The study included 117 patients who attended our clinic for routine outpatient blood tests and who were receiving antiretroviral therapy which included NNRTI or PI. Patients were not informed that drug concentrations were going to be assessed until blood sampling. The time of the last antiretroviral treatment intake and blood sampling were recorded. Drug concentrations were considered optimal if they were above the proposed minimum effective concentration. In addition, efavirenz, nevirapine and atazanavir concentrations were considered potentially toxic if they were higher than 4.0 mg l−1, 6.0 mg l−1, and 0.85 mg l−1, respectively. Results Overall, interindividual variability in NNRTI and PI plasma concentrations was approximately 50%, and only 68.4% of the patients had drug concentrations within the proposed therapeutic range. Inappropriate adherence only explained 35% of subtherapeutic drug concentrations. Conclusion Interindividual variability in trough concentrations of NNRTI and PI among HIV-infected adults is large in routine clinical practice, with drug concentrations being outside the therapeutic window in a significant proportion of patients. Therapeutic drug monitoring may be useful to guide antiretroviral therapy in clinical practice. PMID:17223856

  19. Variability in non-nucleoside reverse transcriptase and protease inhibitor concentrations among HIV-infected adults in routine clinical practice.

    Science.gov (United States)

    Moltó, José; Blanco, Asunción; Miranda, Cristina; Miranda, José; Puig, Jordi; Valle, Marta; DelaVarga, Meritxell; Fumaz, Carmina R; Barbanoj, Manuel José; Clotet, Bonaventura

    2006-11-01

    The objective of this study was to assess interindividual variability in trough concentrations of plasma of non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI) among HIV-infected adults in a routine outpatient setting. One hundred and seventeen patients who attended our clinic for routine blood tests, and who were receiving antiretroviral therapy which included NNRTI or PI were studied. Patients were not informed that drug concentrations were going to be measured until blood sampling. The times of the last antiretroviral dose and of blood sampling were recorded. Drug concentrations were considered optimal if they were above the proposed minimum effective value. In addition, efavirenz, nevirapine and atazanavir concentrations were considered potentially toxic if they were > 4.0 mg l(-1), > 6.0 mg l(-1) and > 0.85 mg l(-1), respectively. Overall, interindividual variability of NNRTI and PI concentrations in plasma was approximately 50%, and only 68.4% of the patients had drug concentrations within the proposed therapeutic range. Poor adherence explained only 35% of subtherapeutic drug concentrations. Interindividual variability in trough concentrations of NNRTI and PI among HIV-infected adults is large in routine clinical practice, with drug concentrations being outside the therapeutic window in a significant proportion of patients. These findings provide further evidence that therapeutic drug monitoring may be useful to guide antiretroviral therapy in clinical practice.

  20. Variability in non-nucleoside reverse transcriptase and protease inhibitors concentrations among HIV-infected adults in routine clinical practice.

    Science.gov (United States)

    Moltó, José; Blanco, Asunción; Miranda, Cristina; Miranda, José; Puig, Jordi; Valle, Marta; Delavarga, Meritxell; Fumaz, Carmina R; Barbanoj, Manuel José; Clotet, Bonaventura

    2007-06-01

    The objective of this study was to assess interindividual variability in plasma trough concentrations of non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI) among HIV-infected adults in an outpatient routine clinical practice setting. The study included 117 patients who attended our clinic for routine outpatient blood tests and who were receiving antiretroviral therapy which included NNRTI or PI. Patients were not informed that drug concentrations were going to be assessed until blood sampling. The time of the last antiretroviral treatment intake and blood sampling were recorded. Drug concentrations were considered optimal if they were above the proposed minimum effective concentration. In addition, efavirenz, nevirapine and atazanavir concentrations were considered potentially toxic if they were higher than 4.0 mg l(-1), 6.0 mg l(-1), and 0.85 mg l(-1), respectively. Overall, interindividual variability in NNRTI and PI plasma concentrations was approximately 50%, and only 68.4% of the patients had drug concentrations within the proposed therapeutic range. Inappropriate adherence only explained 35% of subtherapeutic drug concentrations. Interindividual variability in trough concentrations of NNRTI and PI among HIV-infected adults is large in routine clinical practice, with drug concentrations being outside the therapeutic window in a significant proportion of patients. Therapeutic drug monitoring may be useful to guide antiretroviral therapy in clinical practice.

  1. In Vitro Antioxidant Properties, HIV-1 Reverse Transcriptase and Acetylcholinesterase Inhibitory Effects of Traditional Herbal Preparations Sold in South Africa

    Directory of Open Access Journals (Sweden)

    Johannes Van Staden

    2010-10-01

    Full Text Available The antioxidant potentials for fourteen multipurpose traditional herbal preparations sold in South Africa were determined using the DPPH radical scavenging, ferric reducing power and β-carotene-linoleic acid model system, the anti-HIV-1 reverse transcriptase (RT enzyme inhibitory effects using an ELISA kit and acetylcholinesterase (AChE enzyme inhibition using the microtitre plate assay. Nine of the herbal mixtures (Umzimba omubi, Umuthi wekukhwehlela ne zilonda, Mvusa ukunzi, Umpatisa inkosi, Imbiza ephuzwato, Vusa umzimba, Supreme one hundred, Sejeso herbal mixture Ingwe® and Ingwe® special muti exhibited higher antioxidant potentials, while only four (Imbiza ephuzwato, Ingwe® muthi mixture, Sejeso herbal mixture Ingwe® and African potato extractTM showed potent activity against the RT enzyme. Nine mixtures (Imbiza ephuzwato, Umpatisa inkosi, African potato extractTM, Sejeso herbal mixture Ingwe®, Vusa umzimba; Ingwe® muthi mixture, Ibhubezi™, Lion izifozonke Ingwe® and Ingwe® special muti showed AChE enzyme inhibitory activity greater than 50%. The observed activity exhibited by some of the herbal mixtures gives some credence to the manufacturers’ claims and goes part of the way towards validating their use against certain conditions such as oxidative stress, HIV/AIDS proliferation and some mental conditions. It is however, desirable to carry out further studies to determine the effects of mixing plant species/parts in one mixture on the antioxidant potency as well as isolating active constituents from the herbal mixtures.

  2. Dolutegravir Plus Two Nucleoside Reverse Transcriptase Inhibitors versus Efavirenz Plus Two Nucleoside Reverse Transcriptase Inhibitors As Initial Antiretroviral Therapy for People with HIV: A Systematic Review.

    Directory of Open Access Journals (Sweden)

    George W Rutherford

    Full Text Available Dolutegravir (DTG is a once-daily unboosted second-generation integrase-inhibitor that along with two nucleoside reverse transcriptase inhibitors is one of several regimens recommended by the United States, United Kingdom and European Union for first-line antiretroviral treatment of people with HIV infection. Our objective was to review the evidence for the efficacy and safety of DTG-based first-line regimens compared to efavirenz (EFV-based regimens.We conducted a systematic review. We comprehensively searched a range of databases as well as conference abstracts and a trials registry. We used Cochrane methods in screening and data collection and assessed each study's risk of bias with the Cochrane tool. We meta-analyzed data using a fixed-effects model. We used GRADE to assess evidence quality.From 492 search results, we identified two randomized controlled trials, reported in five peer-reviewed articles and one conference abstract. One trial tested two DTG-based regimens (DTG + abacavir (ABC + lamivudine (3TC or DTG + tenofovir + emtricitabine against an EFV-based regimen (EFV+ ABC+3TC. The other trial tested DTG+ABC+3TC against EFV+ABC+3TC. In meta-analysis, DTG-containing regimens were superior to EFV-containing regimens at 48 weeks and at 96 weeks (RR = 1.10, 95% CI 1.04-1.16; and RR = 1.12, 95% CI 1.04-1.21, respectively. In one trial, the DTG-containing regimen was superior at 144 weeks (RR = 1.13, 95% CI 1.02-1.24. DTG-containing regimens were superior in reducing treatment discontinuation compared to those containing EFV at 96 weeks and at 144 weeks (RR = 0.27, 95% CI 0.15-0.50; and RR = 0.28, 95% CI 0.16-0.48, respectively. Risk of serious adverse events was similar in each regimen at 96 weeks (RR = 1.15, 95% CI 0.80-1.63 and 144 weeks (RR = 0.93, 95% CI 0.68-1.29. Risk of bias was moderate overall, as was GRADE evidence quality.DTG-based regimens should be considered in future World Health Organization guidelines for initial HIV

  3. Development and evaluation of HIV-1 subtype RNA panels for the standardization of HIV-1 NAT assays.

    Science.gov (United States)

    Lee, Sherwin; Wood, Owen; Taffs, Rolf E; Hu, Jinjie; Machuca, Ana; Vallejo, Alejandro; Hewlett, Indira

    2006-11-01

    Multiple nucleic acid-based techniques (NAT) have been implemented for testing blood and plasma donors for HIV-1 RNA which may be detected at an earlier stage of infection when HIV antigen or antibody is absent or below the limit of detection of current assays. The available NAT assays are based on different technologies. In order to evaluate the performance of nucleic acid-based techniques (NAT assays) and to allow accurate comparisons of results from different assays, it is essential to have well characterized specimens with known copy numbers as a standard. For this purpose, a comprehensive study was conducted to develop two HIV-1 RNA reference panels. The first (Panel 1) was prepared using a single specimen from the HIV-1 group M subtype B and consists of panel members with a wide range of HIV-1 RNA copy numbers. Panel 2 consists of 26 members representing HIV-1 group M subtypes A, C, D, E, F, G and groups O and N. For accurate determination of HIV-1 RNA copy numbers of each member of Panel 2, they were analyzed using various testing platforms/technologies available through the cooperation of five independent laboratories participating in the study. A consensus value for HIV RNA copy number was assigned to each member of Panel 2 based on statistical analysis of the data provided by the participants. Both panels could serve as reference panels to be used by manufacturers of HIV NAT tests to evaluate the sensitivity limits of their assays.

  4. Evaluation of TB and HIV services prior to introducing TB-HIV activities in two rural districts in western Kenya

    NARCIS (Netherlands)

    van't Hoog, A. H.; Onyango, J.; Agaya, J.; Akeche, G.; Odero, G.; Lodenyo, W.; Marston, B. J.

    2008-01-01

    SETTING: Health facilities providing tuberculosis (TB) treatment in two districts in rural western Kenya with a high TB and human immunodeficiency virus (HIV) burden. OBJECTIVE: To evaluate TB and HIV/acquired immune-deficiency syndrome (AIDS) services at the facilities and identify barriers to

  5. Evaluation of current rapid HIV test algorithms in Rakai, Uganda.

    Science.gov (United States)

    Galiwango, Ronald M; Musoke, Richard; Lubyayi, Lawrence; Ssekubugu, Robert; Kalibbala, Sarah; Ssekweyama, Viola; Mirembe, Viola; Nakigozi, Gertrude; Reynolds, Steven J; Serwadda, David; Gray, Ronald H; Kigozi, Godfrey

    2013-09-01

    Rapid HIV tests are a crucial component of HIV diagnosis in resource limited settings. In Uganda, the Ministry of Health allows both serial and parallel HIV rapid testing using Determine, Stat-Pak and Uni-Gold. In serial testing, a non-reactive result on Determine ends testing. The performance of serial and parallel algorithms with Determine and Stat-Pak test kits was assessed. A cross-sectional diagnostic test accuracy evaluation using three rapid HIV test kits as per the recommended parallel test algorithm was followed by EIA-WB testing with estimates of the performance of serial testing algorithm. In 2520 participants tested by parallel rapid algorithms, 0.6% had weakly reactive result. Parallel testing had 99.7% sensitivity and 99.8% specificity. If Stat-Pak was used as the first screening test for a serial algorithm, the sensitivity was 99.6% and specificity was 99.7%. However, if Determine was used as the screening test, sensitivity was 97.3% and specificity was 99.9%. Serial testing with Stat-Pak as the initial screening test performed as well as parallel testing, but Determine was a less sensitive screen. Serial testing could be cost saving. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Evaluation Of Algorithms Of Anti- HIV Antibody Tests

    Directory of Open Access Journals (Sweden)

    Paranjape R.S

    1997-01-01

    Full Text Available Research question: Can alternate algorithms be used in place of conventional algorithm for epidemiological studies of HIV infection with less expenses? Objective: To compare the results of HIV sero- prevalence as determined by test algorithms combining three kits with conventional test algorithm. Study design: Cross â€" sectional. Participants: 282 truck drivers. Statistical analysis: Sensitivity and specificity analysis and predictive values. Results: Three different algorithms that do not include Western Blot (WB were compared with the conventional algorithm, in a truck driver population with 5.6% prevalence of HIV â€"I infection. Algorithms with one EIA (Genetic Systems or Biotest and a rapid test (immunocomb or with two EIAs showed 100% positive predictive value in relation to the conventional algorithm. Using an algorithm with EIA as screening test and a rapid test as a confirmatory test was 50 to 70% less expensive than the conventional algorithm per positive scrum sample. These algorithms obviate the interpretation of indeterminate results and also give differential diagnosis of HIV-2 infection. Alternate algorithms are ideally suited for community based control programme in developing countries. Application of these algorithms in population with low prevalence should also be studied in order to evaluate universal applicability.

  7. Transgender HIV prevention: implementation and evaluation of a workshop.

    Science.gov (United States)

    Bockting, W O; Rosser, B R; Scheltema, K

    1999-04-01

    Virtually no HIV prevention education has specifically targeted the transgender community. To fill this void, a transgender HIV prevention workshop was developed, implemented and evaluated. A 4 h workshop, grounded in the Health Belief Model and the Eroticizing Safer Sex approach, combined lectures, videos, a panel, discussion, roleplay and exercises. Evaluation using a pre-, post- and follow-up test design showed an increase in knowledge and an initial increase in positive attitudes that diminished over time. Due to the small sample size (N = 59) and limited frequency of risk behavior, a significant decrease in unsafe sexual or needle practices could not be demonstrated. However, findings suggested an increase in safer sexual behaviors such as (mutual) masturbation. Peer support improved significantly. Future prevention education should make special efforts to target the more difficult-to-reach, high-risk subgroups of the transgender population.

  8. Use of nucleoside reverse transcriptase inhibitors and risk of myocardial infarction in HIV-infected patients

    NARCIS (Netherlands)

    Lundgren, Jens D.; Neuhaus, Jacqueline; Babiker, Abdel; Cooper, David; Duprez, Daniel; El-Sadr, Wafaa; Emery, Sean; Gordin, Fred; Kowalska, Justyna; Phillips, Andrew; Prineas, Ronald J.; Reiss, Peter; Sabin, Caroline; Tracy, Russell; Weber, Rainer; Grund, Birgit; Neaton, James D.

    2008-01-01

    BACKGROUND: Two nucleos(t)ide reverse transcriptase inhibitors (NRTIs)--abacavir and didanosine--may each be associated with excess risk of myocardial infarction. The reproducibility of this finding in an independent dataset was explored and plausible biological mechanisms were sought. METHODS:

  9. Structural features in the HIV-1 repeat region facilitate strand transfer during reverse transcription

    NARCIS (Netherlands)

    Berkhout, B.; Vastenhouw, N. L.; Klasens, B. I.; Huthoff, H.

    2001-01-01

    Two obligatory DNA strand transfers take place during reverse transcription of a retroviral RNA genome. The first strand transfer is facilitated by terminal repeat (R) elements in the viral genome. This strand-transfer reaction depends on base pairing between the cDNA of the 5'R and the 3'R. There

  10. HIV/AIDS/STD prevention intervention messages: An evaluation of ...

    African Journals Online (AJOL)

    The aim of this study is to evaluate HIV/AIDS/STD prevention intervention messages on a rural adult (25-49 years) sample in South Africa over a period of 15 months. A representative community sample of 398 adults at time 1 and 382 at time 2 (25-49 years) participated in the study using a three-stage cluster sampling ...

  11. Evaluation of the pharmacotherapeutic profile of HIV/AIDS bearers

    OpenAIRE

    Bessa Filho, Valter; Bezerra, Everton R.; Ferreira, Aluísio M.; Janebro, Daniele I.; Queiroz, Maria do S.R.; Monteiro, Matheus M. O.; Santana, Jorge E.G.; Vicente, Carlos H.T.B.

    2011-01-01

    In this work was evaluated the pharmacotherapeutic profile of HIV/AIDS bearers registered in the Service of Specialized Attendance located in Campina Grande–PB, Brazil. The research was of the type traverse, documental, descriptive and analytical and was realized in the period of August to October 2010. Were appraised the patient records of 188 people being 66 % males and 34% females. The most part (36 %) just studied the education fundamental level and presented age group between 40-49...

  12. Evaluation of the Bio-Rad Geenius HIV-1/2 test as a confirmatory assay.

    Science.gov (United States)

    Montesinos, Isabel; Eykmans, Joelle; Delforge, Marie-Luce

    2014-08-01

    We have evaluated the recently Conformité Européenne (CE)-marked Bio-Rad Geenius human immunodeficiency virus (HIV)1/2 as a rapid and simple alternative to western blot for confirmation of HIV screening results. A total of 160 serum samples were tested: 44 HIV-1 reactive samples by a fourth-generation Murex HIV Ag/Ab and/or Vidas HIV Duo Ultra, five HIV-2 reactive samples, 15 HIV-1 non-B subtype samples and 11 confirmed HIV-1 early seroconversion samples, 72 nonreactive samples, eight indeterminate samples by MP HIV BLOT 2.2 confirmed negative after follow-up and five low-reactive samples by enzyme immunoassay (EIA) negative by MP HIV BLOT 2.2. The samples were tested according to the manufacturer's guidelines. The overall sensitivity for Bio-Rad Geenius HIV1/2 assay was 92%. Five out of 11 early seroconversion samples were tested positive, four negative and two indeterminate. All HIV-1 non-B subtype samples were tested positive. Two out of the five HIV-2 reactive samples were tested positive HIV-2, two positive HIV-2 with HIV-1 cross-reaction and one HIV positive untypable. After excluding early seroconversion samples, the sensitivity of Bio-Rad Geenius HIV1/2 assay reached 100%. Overall specificity was 96%. All HIV negative serums by fourth-generation EIA were tested negative. All five low-reactive samples by EIA, negative by HIV BLOT 2.2 were tested negative by Bio-Rad Geenius HIV1/2. Two out of the eight indeterminate samples by MP HIV BLOT 2.2 that were confirmed negative after follow-up were tested indeterminate and one invalid, the other five were negative. After excluding these last 13 samples, the specificity of Bio-Rad Geenius HIV1/2 assay reached 100%. In comparison with MP HIV BLOT 2.2, the Bio-Rad Geenius HIV1/2 assay was markedly time saving, allowed full traceability, automatic reading and interpretation. The Bio-Rad Geenius HIV1/2 confirmatory system represents a reliable alternative to other confirmatory assays in HIV testing algorithms and

  13. Identification of the critical sites of NNRTI-resistance in reverse transcriptase of HIV-1 CRF_BC strains.

    Directory of Open Access Journals (Sweden)

    Yang Huang

    Full Text Available BACKGROUND: The polymorphisms involved in drug resistance to non-nucleoside reverse transcriptase inhibitors (NNRTIs in HIV-1 CRF_BC, the most prevalent HIV-1 strain in China, have been poorly characterized. RESULTS: To reveal the drug resistance mutations, we compared the gene sequences of pol region of HIV-1 CRF_BC from 631 treatment-naïve and 363 treatment-experienced patients using the selection pressure-based method. We calculated an individual Ka/Ks value for each specific amino acid mutation. Result showed that eight polymorphic mutations (W88C, K101Q, I132L, R135L, T139K/R, H221Y and L228R in RT for treatment-experienced patients were identified, while they, except for R135L, were completely absent in those from treatment-naïve patients. The I132L and T139K/R mutants exhibited high-level resistance to DLV and NVP and moderate resistance to TMC-125 and EFV, while the K101Q and H221Y mutants exhibited an increased resistance to all four NNRTIs tested. The W88C, R135L, and L228R may be RTI-induced adaptive mutations. Y181C+K101Q mutant showed a 2.5-, 4.4-, and 4.7-fold higher resistance to TMC-125, NVP and EFV, respectively, than Y181C alone mutant, while Y181C+H221Y or K103N+H221Y mutants had significantly higher resistance to all four NNRTIs than Y181C or K103N mutants. K103N+T139K and G190A+T139K mutant induce higher resistance (2.0∼14.2-fold and 1.5∼7.2-fold, respectively to all four NNRTIs than K103N or G190A alone mutation. CONCLUSIONS: I132L and T139K/R are rare but critical mutations associated with NNRTI-resistance for some NNRTIs. K101Q, H221Y and T139K can enhance K103N/Y181C/G190A-assocated NNRTI-resistance. Monitoring these mutations will provide useful information for rational design of the NNRTI-based antiretroviral regimen for HIV-1 CRF_BC-infected patients.

  14. Evaluating Reverse Supply Chain Efficiency: Manufacturer’s Perspective

    Directory of Open Access Journals (Sweden)

    Manasvi Kumar

    2014-01-01

    Full Text Available The paper aims to illustrate the use of fuzzy data envelopment analysis (DEA in analyzing reverse supply chain (RSC performance from the manufacturer’s perspective. By using an alternative α-cut approach, the fuzzy DEA model was converted into a crisp linear programming problem, thereby altering the problem to an interval programming one. The model is able to obtain precise and robust efficiency values. An investigation was also made between the obtained efficiency scores and certain relevant background information of the companies. The study revealed that while ISO 14001 certification usually ensures an environmentally friendly supply chain network, companies which have implemented RSC techniques since a longer duration do not necessarily have a more efficient supply chain in general.

  15. Evaluation of an HIV-risk reduction programme for first-year ...

    African Journals Online (AJOL)

    Evaluation of an HIV-risk reduction programme for first-year university students in South Africa. ... African Journal for Physical Activity and Health Sciences. Journal Home ... Results indicated that HIV-related knowledge; condom knowledge and risk perception were enhanced by the HIV- related risk reduction programme.

  16. Evaluation of an HIV/AIDS peer education programme in a South ...

    African Journals Online (AJOL)

    Objectives. To evaluate a South African workplace HIV I AIDS peer-education programme running since 1997. Methods. In 2001 a cross-sectional study was done of 900 retail-section employees in three geographical areas. The study measured HIV I AIDS knowledge, attitudes towards people living with HIV I AIDS, belief ...

  17. Nondestructive evaluation for remanent life of aged 12Cr ferrite heat resisting steel by reversible permeability

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Kwon-Sang, E-mail: ryuks@kriss.re.kr [Korea Research Institute of Standards and Science, Centre for Materials Measurement, Doryong Dong, Yuseong-Gu, Daejeon 305-340 (Korea, Republic of); Kim, Chung-Seok [Automotive Engineering, Hanyang University, Seoul 133-791 (Korea, Republic of); Baek, Un-bong [Korea Research Institute of Standards and Science, Centre for Materials Measurement, Doryong Dong, Yuseong-Gu, Daejeon 305-340 (Korea, Republic of); Lee, Jong Suk [Gangneung-Wonju National University, Wonju 220-711 (Korea, Republic of)

    2013-01-15

    We present a magnetic and nondestructive method to evaluate the remanent life of advanced ferritic steel using the value of reversible permeability. The method to measure reversible permeability is based on the theory that the value of reversible permeability is the same differential of the hysteresis loop. The measurement principle is based on the foundation of harmonics voltage induced in a sensing coil using a lock-in amplifier tuned to the frequency of the exciting one. Results obtained for reversible permeability, Vickers hardness, and tensile strength on the aged samples show that the peak interval of reversible permeability, Vickers hardness and tensile strength decrease as aging time increases. We could estimate the remanent life of advanced ferritic steel by using the relationship between the peak interval of reversible permeability and the Larson-Miller parameter, non-destructively. - Highlights: Black-Right-Pointing-Pointer Magnetic, nondestructive evaluation method of remanent life of 12Cr ferritic steel is presented. Black-Right-Pointing-Pointer Peak interval of reversible permeability decreases with the increase of aging time. Black-Right-Pointing-Pointer Mechanical properties decrease with the increase of aging time. Black-Right-Pointing-Pointer Magnetic and mechanical properties are decreased with increase of Larson-Miller parameter. Black-Right-Pointing-Pointer Reversible permeability is nondestructively used to estimate remanent life of 12Cr ferrite steel.

  18. The case for addressing primary resistance mutations to non-nucleoside reverse transcriptase inhibitors to treat children born from mothers living with HIV in sub-Saharan Africa

    OpenAIRE

    Khady Kébé; Laurent Bélec; Halimatou Diop Ndiaye; Sokhna Bousso Gueye; Abou Abdallah Malick Diouara; Safiétou Ngom; Ndéye Rama Diagne Gueye; Ngagne Mbaye; Haby Signaté Sy; Souleymane Mboup; Coumba Touré Kane

    2014-01-01

    The prevalence of human immunodeficiency virus (HIV) drug resistance mutations (DRMs) was estimated in 25 untreated infants who were living with HIV-1, younger than 13 months and living in Senegal. Antiretroviral DRMs were detected in 8 of 25 (32%) children. Non-nucleoside reverse transcriptase inhibitor (NNRTI) DRMs were present in all (100%) children whose viruses harboured DRMs: K103N in 43%; Y181C, K101E and V106M each in 29%; and Y188L in 14%. The D67N thymidine-analogue mutation was obs...

  19. Identification of a novel resistance (E40F and compensatory (K43E substitution in HIV-1 reverse transcriptase

    Directory of Open Access Journals (Sweden)

    Kagan Ron M

    2008-02-01

    Full Text Available Abstract Background HIV-1 nucleoside reverse transcriptase inhibitors (NRTIs have been used in the clinic for over twenty years. Interestingly, the complete resistance pattern to this class has not been fully elucidated. Novel mutations in RT appearing during treatment failure are still being identified. To unravel the role of two of these newly identified changes, E40F and K43E, we investigated their effect on viral drug susceptibility and replicative capacity. Results A large database (Quest Diagnostics database was analysed to determine the associations of the E40F and K43E changes with known resistance mutations. Both amino acid changes are strongly associated with the well known NRTI-resistance mutations M41L, L210W and T215Y. In addition, a strong positive association between these changes themselves was observed. A panel of recombinant viruses was generated by site-directed mutagenesis and phenotypically analysed. To determine the effect on replication capacity, competition and in vitro evolution experiments were performed. Introduction of E40F results in an increase in Zidovudine resistance ranging from nine to fourteen fold depending on the RT background and at the same time confers a decrease in viral replication capacity. The K43E change does not decrease the susceptibility to Zidovudine but increases viral replication capacity, when combined with E40F, demonstrating a compensatory role for this codon change. Conclusion In conclusion, we have identified a novel resistance (E40F and compensatory (K43E change in HIV-1 RT. Further research is indicated to analyse the clinical importance of these changes.

  20. Effects of HIV-1 reverse transcriptase connection subdomain mutations on polypurine tract removal and initiation of (+)-strand DNA synthesis.

    Science.gov (United States)

    Betancor, Gilberto; Álvarez, Mar; Marcelli, Barbara; Andrés, Cristina; Martínez, Miguel A; Menéndez-Arias, Luis

    2015-02-27

    HIV-1 reverse transcriptase (RT) connection subdomain mutations at positions 348, 369 and 376 have been associated with resistance to non-nucleoside RT inhibitors (NNRTIs). N348I may interfere with the initiation of (+)-strand DNA synthesis by reducing polypurine tract (PPT) removal in the presence of nevirapine. The effect of NNRTIs on the RNase H-mediated cleavage of PPT-containing template-primers has been studied with wild-type HIV-1 RT and mutants N348I, T369I, T369V, T376S and N348I/T369I. In the presence of NNRTIs, all RTs were able to stimulate PPT cleavage after primer elongation. The enhancing effects of nevirapine and efavirenz were reduced in RTs carrying mutation N348I, and specially N348I/T369I. However, those mutations had no effect on rilpivirine-mediated cleavage. Prior to elongation, the PPT remains resilient to cleavage, although efavirenz and rilpivirine facilitate RNase H-mediated trimming of its 3'-end. The integrity of the 3'-end is essential for the initiation of (+)-strand DNA synthesis. In the presence of dNTPs, rilpivirine was the most effective inhibitor of (+)-strand DNA synthesis blocking nucleotide incorporation and preventing usage of available PPT primers. The N348I/T369I RT showed reduced ability to generate short RNA products revealing a cleavage window defect. Its lower RNase H activity could be attributed to enhanced rigidity compared to the wild-type enzyme. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  1. Distinctive Drug-resistant Mutation Profiles and Interpretations of HIV-1 Proviral DNA Revealed by Deep Sequencing in Reverse Transcriptase.

    Science.gov (United States)

    Yin, Qian Qian; Li, Zhen Peng; Zhao, Hai; Pan, Dong; Wang, Yan; Xu, Wei Si; Xing, Hui; Feng, Yi; Jiang, Shi Bo; Shao, Yi Ming; Ma, Li Ying

    2016-04-01

    To investigate distinctive features in drug-resistant mutations (DRMs) and interpretations for reverse transcriptase inhibitors (RTIs) between proviral DNA and paired viral RNA in HIV-1-infected patients. Forty-three HIV-1-infected individuals receiving first-line antiretroviral therapy were recruited to participate in a multicenter AIDS Cohort Study in Anhui and Henan Provinces in China in 2004. Drug resistance genotyping was performed by bulk sequencing and deep sequencing on the plasma and whole blood of 77 samples, respectively. Drug-resistance interpretation was compared between viral RNA and paired proviral DNA. Compared with bulk sequencing, deep sequencing could detect more DRMs and samples with DRMs in both viral RNA and proviral DNA. The mutations M184I and M230I were more prevalent in proviral DNA than in viral RNA (Fisher's exact test, PDNA, and 5 of these samples with different DRMs between proviral DNA and paired viral RNA showed a higher level of drug resistance to the first-line drugs. Considering 'minority resistant variants', 22 samples (28.57%) were associated with a higher level of drug resistance to the tested RTIs for proviral DNA when compared with paired viral RNA. Compared with viral RNA, the distinctive information of DRMs and drug resistance interpretations for proviral DNA could be obtained by deep sequencing, which could provide more detailed and precise information for drug resistance monitoring and the rational design of optimal antiretroviral therapy regimens. Copyright © 2016 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  2. Parameterization of AZT-A widely used nucleoside inhibitor of HIV-1 reverse transcriptase

    Science.gov (United States)

    Carvalho, Alexandra T. P.; Fernandes, Pedro A.; Ramos, Maria J.

    Seven nucleoside reverse transcriptase (RT) inhibitors are currently used in the clinical treatment of acquired immunodeficiency syndrome (AIDS). These substrate analogues block DNA synthesis by the viral enzyme RT. However, the emergence of resistant variants of RT allied to their long-term toxicity requires the design of new and better RT inhibitors, with long-term in vivo efficacy. In this work we used density functional theory (DFT) calculations to develop a set of molecular mechanics (MM) parameters committed to the AMBER force field for one of the most used in the clinic nucleoside reverse transcriptase inhibitors (NRTIs): zidovudine (AZT). These parameters were tested by comparing the optimized geometries of AZT at both the DFT and MM levels of theory. The ability of the new parameters to reproduce the torsional energy of the azide group was also verified by scanning the surface in MM with the new parameters and comparing the results with the same potential energy surface (PES) at the DFT level. Finally, the parameters were validated through classical MD simulations of AZT in aqueous environment.

  3. Evaluation of the automated 'Enzymen-Test Anti HIV-1 + 2' and 'Enzymen-Test Anti HIV-1/2 selective' for the combined detection and differentiation of anti-HIV-1 and anti-HIV-2 antibodies.

    Science.gov (United States)

    Weber, B; Hess, G; Koberstein, R; Doerr, H W

    1993-10-01

    A new, modular automated ELISA (test 1) for HIV-1 and HIV-2 antibody detection and differentiation (Enzymun-Test Anti HIV-1 + 2; anti HIV 1/2 selective, Boehringer Mannheim) was compared with 3 alternative enzyme immunoassays (Abbott recombinant HIV-1/HIV-2 3rd generation EIA, Abbott (test 2); Enzygnost HIV 1 + 2, Behringwerke (test 3); and Wellcozyme HIV recombinant, Murex (test 4)) and Western blot (New LAV I Blot and New LAV II Blot; Diagnostics Pasteur). 380 serum samples from HIV-1 and HIV-2 seropositive patients at different stages of disease, high risk individuals, patients with conditions unrelated to AIDS and from healthy blood donors were used in this evaluation along with 6 seroconversion panels, 6 serum dilution series and 'tricky' sera (repeatedly positive results in ELISA, but negative or undeterminate in Western blot; n = 67). Using the Western blot as reference assay, the overall sensitivity of the four ELISAs was 100%. Test 4 showed the highest sensitivity for antibody detection in seroconversion and dilution series. A high specificity was achieved with test 1 (100%) and test 2 (99.4%). A relatively high rate of false positive results were obtained with test 2 (n = 12) and test 3 (n = 10) by testing 'tricky' sera or samples obtained from healthy blood donors. In comparison to Western blot, a clear differentiation between HIV-1 and HIV-2 antibody serum samples was achieved with the Enzymun-Test. The results of the present study show that the Enzymun-Test provides reliable selective HIV-1 and HIV-2 antibody detection at a cost which is significantly lower than the costs of Western blot tests. Furthermore, the evaluation of test 1 suggests, that it is a highly specific assay for HIV antibody detection.

  4. Structural Insights into HIV Reverse Transcriptase Mutations Q151M and Q151M Complex That Confer Multinucleoside Drug Resistance

    Energy Technology Data Exchange (ETDEWEB)

    Das, Kalyan; Martinez, Sergio E.; Arnold, Eddy

    2017-04-10

    HIV-1 reverse transcriptase (RT) is targeted by multiple drugs. RT mutations that confer resistance to nucleoside RT inhibitors (NRTIs) emerge during clinical use. Q151M and four associated mutations, A62V, V75I, F77L, and F116Y, were detected in patients failing therapies with dideoxynucleosides (didanosine [ddI], zalcitabine [ddC]) and/or zidovudine (AZT). The cluster of the five mutations is referred to as the Q151M complex (Q151Mc), and an RT or virus containing Q151Mc exhibits resistance to multiple NRTIs. To understand the structural basis for Q151M and Q151Mc resistance, we systematically determined the crystal structures of the wild-type RT/double-stranded DNA (dsDNA)/dATP (complex I), wild-type RT/dsDNA/ddATP (complex II), Q151M RT/dsDNA/dATP (complex III), Q151Mc RT/dsDNA/dATP (complex IV), and Q151Mc RT/dsDNA/ddATP (complex V) ternary complexes. The structures revealed that the deoxyribose rings of dATP and ddATP have 3'-endo and 3'-exo conformations, respectively. The single mutation Q151M introduces conformational perturbation at the deoxynucleoside triphosphate (dNTP)-binding pocket, and the mutated pocket may exist in multiple conformations. The compensatory set of mutations in Q151Mc, particularly F116Y, restricts the side chain flexibility of M151 and helps restore the DNA polymerization efficiency of the enzyme. The altered dNTP-binding pocket in Q151Mc RT has the Q151-R72 hydrogen bond removed and has a switched conformation for the key conserved residue R72 compared to that in wild-type RT. On the basis of a modeled structure of hepatitis B virus (HBV) polymerase, the residues R72, Y116, M151, and M184 in Q151Mc HIV-1 RT are conserved in wild-type HBV polymerase as residues R41, Y89, M171, and M204, respectively; functionally, both Q151Mc HIV-1 and wild-type HBV are resistant to dideoxynucleoside analogs.

  5. N348I in the connection domain of HIV-1 reverse transcriptase confers zidovudine and nevirapine resistance.

    Directory of Open Access Journals (Sweden)

    Soo-Huey Yap

    2007-12-01

    Full Text Available The catalytically active 66-kDa subunit of the human immunodeficiency virus type 1 (HIV-1 reverse transcriptase (RT consists of DNA polymerase, connection, and ribonuclease H (RNase H domains. Almost all known RT inhibitor resistance mutations identified to date map to the polymerase domain of the enzyme. However, the connection and RNase H domains are not routinely analysed in clinical samples and none of the genotyping assays available for patient management sequence the entire RT coding region. The British Columbia Centre for Excellence in HIV/AIDS (the Centre genotypes clinical isolates up to codon 400 in RT, and our retrospective statistical analyses of the Centre's database have identified an N348I mutation in the RT connection domain in treatment-experienced individuals. The objective of this multidisciplinary study was to establish the in vivo relevance of this mutation and its role in drug resistance.The prevalence of N348I in clinical isolates, the time taken for it to emerge under selective drug pressure, and its association with changes in viral load, specific drug treatment, and known drug resistance mutations was analysed from genotypes, viral loads, and treatment histories from the Centre's database. N348I increased in prevalence from below 1% in 368 treatment-naïve individuals to 12.1% in 1,009 treatment-experienced patients (p = 7.7 x 10(-12. N348I appeared early in therapy and was highly associated with thymidine analogue mutations (TAMs M41L and T215Y/F (p < 0.001, the lamivudine resistance mutations M184V/I (p < 0.001, and non-nucleoside RTI (NNRTI resistance mutations K103N and Y181C/I (p < 0.001. The association with TAMs and NNRTI resistance mutations was consistent with the selection of N348I in patients treated with regimens that included both zidovudine and nevirapine (odds ratio 2.62, 95% confidence interval 1.43-4.81. The appearance of N348I was associated with a significant increase in viral load (p < 0.001, which

  6. An Evaluation of Introduction of Rapid HIV Testing in a Perinatal Program.

    Science.gov (United States)

    Saunders, Sarah; Tulloch, Karen; Maan, Evelyn J; van Schalkwyk, Julianne; Money, Deborah M

    2017-08-01

    This study was conducted to evaluate the roll-out of rapid HIV testing as part of an emergency Prevention of Perinatal HIV Transmission Program. Specifically, HIV prevalence in this population, the reason(s) for performing the rapid HIV test, and compliance with recommendations for antiretroviral prophylaxis were assessed. Since November 2011, all women presenting to a tertiary labour and delivery unit with unknown HIV status or with ongoing risk of HIV infection since their last HIV test were offered rapid HIV testing. Through retrospective chart review, demographic data, HIV risk and prior testing history, and antiretroviral prophylaxis, data were collected and descriptive statistics were performed. One hundred fourteen rapid HIV tests were conducted and there were two preliminary reactive rapid results (one true positive, one false positive). None of the infants was HIV infected. Sixty-three percent of women had multiple risk factors for HIV acquisition, most commonly intravenous drug use (54%). Forty-four percent of women were within the 4-week seroconversion window at the time of delivery; 25% of these women and 52% of their infants received prophylactic drug therapy. Rapid HIV testing identified a high-risk cohort and enabled aggressive management of a newly diagnosed HIV-positive pregnancy, successfully preventing perinatal HIV transmission. Risk factors for HIV acquisition were ongoing within the seroconversion window for over half of the women, impacting the utility of the test in eliminating unnecessary antiretroviral prophylaxis in this population because prophylaxis is recommended despite a negative rapid HIV test in these cases. Copyright © 2017 The Society of Obstetricians and Gynaecologists of Canada/La Société des obstétriciens et gynécologues du Canada. Published by Elsevier Inc. All rights reserved.

  7. Economic and Environmental Advantage Evaluation of the Reverse Logistic Implementation in the Supermarket Retail

    OpenAIRE

    Oliveira Neto, Geraldo,; Sousa, Washington,

    2014-01-01

    Part 1: Knowledge-Based Sustainability; International audience; In this article it is presented a market study case that allowed a recycling specialized organization with physical space to implement the reverse logistic of the following packing residues: cardboard, Styrofoam, plastic and aluminum into correct residue destination. This study’s objective consists in developing a methodology to evaluate economic and environmental advantage of the reverse logistic implementation in the supermarke...

  8. A protein with antiproliferative, antifungal and HIV-1 reverse transcriptase inhibitory activities from caper (Capparis spinosa) seeds.

    Science.gov (United States)

    Lam, Sze-Kwan; Ng, Tzi-Bun

    2009-05-01

    A protein exhibiting an N-terminal amino acid sequence with some similarity to imidazoleglycerol phosphate synthase was purified from fresh Capparis spinosa melon seeds. The purification protocol entailed anion exchange chromatography on DEAE-cellulose, cation exchange chromatography on SP-Sepharose, and finally gel filtration by fast protein liquid chromatography on Superdex 75. The protein was adsorbed using 20 mM Tris-HCl buffer (pH 7.4) and desorbed using 1 M NaCl in the starting buffer from the DEAE-cellulose column and SP-Sepharose column. The protein demonstrated a molecular mass of 38 kDa in gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that it was monomeric. The protein inhibited proliferation of hepatoma HepG2 cells, colon cancer HT29 cells and breast cancer MCF-7 cells with an IC(50) of about 1, 40 and 60 microM, respectively. It inhibited HIV-1 reverse transcriptase with IC(50) of 0.23 microM. It inhibited mycelial growth in the fungus, Valsa mali. It did not exhibit hemagglutinating, ribonuclease, mitogenic or protease inhibitory activities.

  9. A trypsin inhibitor from rambutan seeds with antitumor, anti-HIV-1 reverse transcriptase, and nitric oxide-inducing properties.

    Science.gov (United States)

    Fang, Evandro Fei; Ng, Tzi Bun

    2015-04-01

    Nephelium lappaceum L., commonly known as "rambutan," is a typical tropical tree and is well known for its juicy and sweet fruit which has an exotic flavor. Chemical studies on rambutan have led to the identification of various components such as monoterpene lactones and volatile compounds. Here, a 22.5-kDa trypsin inhibitor (N . lappaceum trypsin inhibitor (NLTI)) was isolated from fresh rambutan seeds using liquid chromatographical techniques. NLTI reduced the proteolytic activities of both trypsin and α-chymotrypsin. Dithiothreitol reduced the trypsin inhibitory activity of NLTI at a concentration of 1 mM, indicating that an intact disulfide bond is essential to the activity. NLTI inhibited HIV-1 reverse transcriptase with an IC50 of 0.73 μM. In addition, NLTI manifested a time- and dose-dependent inhibitory effect on growth in many tumor cells. NLTI is one of the few trypsin inhibitors with nitric oxide-inducing activity and may find application in tumor therapy.

  10. A Novel Laccase with Potent Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Mycelia of Mushroom Coprinus comatus

    Directory of Open Access Journals (Sweden)

    Shuang Zhao

    2014-01-01

    Full Text Available A novel laccase was isolated and purified from fermentation mycelia of mushroom Coprinus comatus with an isolation procedure including three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose and one gel-filtration step by fast protein liquid chromatography on Superdex 75. The purified enzyme was a monomeric protein with a molecular weight of 64 kDa. It possessed a unique N-terminal amino acid sequence of AIGPVADLKV, which has considerably high sequence similarity with that of other fungal laccases, but is different from that of C. comatus laccases reported. The enzyme manifested an optimal pH value of 2.0 and an optimal temperature of 60°C using 2,2′-azinobis(3-ethylbenzothiazolone-6-sulfonic acid diammonium salt (ABTS as the substrate. The laccase displayed, at pH 2.0 and 37°C, Km values of 1.59 mM towards ABTS. It potently suppressed proliferation of tumor cell lines HepG2 and MCF7, and inhibited human immunodeficiency virus type 1 (HIV-1 reverse transcriptase (RT with an IC50 value of 3.46 μM, 4.95 μM, and 5.85 μM, respectively, signifying that it is an antipathogenic protein.

  11. A novel laccase with potent antiproliferative and HIV-1 reverse transcriptase inhibitory activities from mycelia of mushroom Coprinus comatus.

    Science.gov (United States)

    Zhao, Shuang; Rong, Cheng-Bo; Kong, Chang; Liu, Yu; Xu, Feng; Miao, Qian-Jiang; Wang, Shou-Xian; Wang, He-Xiang; Zhang, Guo-Qing

    2014-01-01

    A novel laccase was isolated and purified from fermentation mycelia of mushroom Coprinus comatus with an isolation procedure including three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose and one gel-filtration step by fast protein liquid chromatography on Superdex 75. The purified enzyme was a monomeric protein with a molecular weight of 64 kDa. It possessed a unique N-terminal amino acid sequence of AIGPVADLKV, which has considerably high sequence similarity with that of other fungal laccases, but is different from that of C. comatus laccases reported. The enzyme manifested an optimal pH value of 2.0 and an optimal temperature of 60°C using 2,2'-azinobis(3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) as the substrate. The laccase displayed, at pH 2.0 and 37°C, K(m) values of 1.59 mM towards ABTS. It potently suppressed proliferation of tumor cell lines HepG2 and MCF7, and inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) with an IC50 value of 3.46 μM, 4.95 μM, and 5.85 μM, respectively, signifying that it is an antipathogenic protein.

  12. Improving fold resistance prediction of HIV-1 against protease and reverse transcriptase inhibitors using artificial neural networks.

    Science.gov (United States)

    Sheik Amamuddy, Olivier; Bishop, Nigel T; Tastan Bishop, Özlem

    2017-08-15

    Drug resistance in HIV treatment is still a worldwide problem. Predicting resistance to antiretrovirals (ARVs) before starting any treatment is important. Prediction accuracy is essential, as low-accuracy predictions increase the risk of prescribing sub-optimal drug regimens leading to patients developing resistance sooner. Artificial Neural Networks (ANNs) are a powerful tool that would be able to assist in drug resistance prediction. In this study, we constrained the dataset to subtype B, sacrificing generalizability for a higher predictive performance, and demonstrated that the predictive quality of the ANN regression models have definite improvement for most ARVs. Trained regression ANNs were optimized for eight protease inhibitors, six nucleoside reverse transcriptase (RT) inhibitors and four non-nucleoside RT inhibitors by experimenting combinations of rare variant filtering (none versus 1 residue occurrence) and ANN topologies (1-3 hidden layers with 2, 4, 6, 8 and 10 nodes per layer). Single hidden layers (5-20 nodes) were used for training where overfitting was detected. 5-fold cross-validation produced mean R2 values over 0.95 and standard deviations lower than 0.04 for all but two antiretrovirals. Overall, higher accuracies and lower variances (compared to results published in 2016) were obtained by experimenting with various preprocessing methods, while focusing on the most prevalent subtype in the raw dataset (subtype B).We thus highlight the need to develop and make available subtype-specific datasets for developing higher accuracy in drug-resistance prediction methods.

  13. Potent Inhibition of HIV-1 Reverse Transcriptase and Replication by Nonpseudoknot, “UCAA-motif” RNA Aptamers

    Directory of Open Access Journals (Sweden)

    Angela S Whatley

    2013-01-01

    Full Text Available RNA aptamers that bind the reverse transcriptase (RT of human immunodeficiency virus (HIV compete with nucleic acid primer/template for access to RT, inhibit RT enzymatic activity in vitro, and suppress viral replication when expressed in human cells. Numerous pseudoknot aptamers have been identified by sequence analysis, but relatively few have been confirmed experimentally. In this work, a screen of nearly 100 full-length and >60 truncated aptamer transcripts established the predictive value of the F1Pk and F2Pk pseudoknot signature motifs. The screen also identified a new, nonpseudoknot motif with a conserved unpaired UCAA element. High-throughput sequence (HTS analysis identified 181 clusters capable of forming this novel element. Comparative sequence analysis, enzymatic probing and RT inhibition by aptamer variants established the essential requirements of the motif, which include two conserved base pairs (AC/GU on the 5′ side of the unpaired UCAA. Aptamers in this family inhibit RT in primer extension assays with IC50 values in the low nmol/l range, and they suppress viral replication with a potency that is comparable with that of previously studied aptamers. All three known anti-RT aptamer families (pseudoknots, the UCAA element, and the recently described “(6/5AL” motif are therefore suitable for developing aptamer-based antiviral gene therapies.

  14. Inhibition of HIV-1 by Non-Nucleoside Reverse Transcriptase Inhibitors via an Induced Fit Mechanism - Importance of Slow Dissociation and Relaxation Rates for Antiviral Efficacy

    OpenAIRE

    Elinder, Malin; Selhorst, Philippe; Vanham, Guido; Öberg, Bo; Vrang, Lotta; Danielson, U. Helena

    2010-01-01

    Abstract The importance of slow dissociation of non-nucleoside reverse transcriptase inhibitors (NNRTI) for antiviral effect has been investigated. The interaction kinetic characteristics of a series of NNRTIs and wild type and drug resistant variants of HIV-1 RT (EC 2.7.7.49) were analyzed by SPR biosensor technology. The antiviral effect was determined in MT-4 and peripheral blood mononuclear cells. Due to extremely slow dissociation rates and a complex interaction mechanism, rat...

  15. Measuring HIV self-management in women living with HIV/AIDS: a psychometric evaluation study of the HIV Self-management Scale.

    Science.gov (United States)

    Webel, Allison R; Asher, Alice; Cuca, Yvette; Okonsky, Jennifer G; Kaihura, Alphoncina; Dawson Rose, Carol; Hanson, Jan E; Salata, Robert A; Burant, Christopher J

    2012-07-01

    To develop and validate the HIV Self-management Scale for women, a new measure of HIV self-management, defined as the day-to-day decisions that individuals make to manage their illness. The development and validation of the scale was undertaken in 3 phases: focus groups, expert review, and psychometric evaluation. Focus groups identified items describing the process and context of self-management in women living with HIV/AIDS (WLHA). Items were refined using expert review and were then administered to WLHA in 2 sites in the United States (n = 260). Validity of the scale was assessed through factor analyses, model fit statistics, reliability testing, and convergent and discriminate validity. The final scale consists of 3 domains with 20 items describing the construct of HIV self-management. Daily self-management health practices, social support and HIV self-management, and chronicity of HIV self-management comprise the 3 domains. These domains explained 48.6% of the total variance in the scale. The item mean scores ranged from 1.7 to 2.8, and each domain demonstrated acceptable reliability (0.72-0.86) and stability (0.61-0.85). Self-management is critical for WLHA, who constitute over 50% of people living with HIV/AIDS (PLWHA) and have poorer health outcomes than their male counterparts. Methods to assess the self-management behavior of WLHA are needed to enhance their health and wellbeing. Presently, no scales exist to measure HIV self-management. Our new 20-item HIV Self-management Scale is a valid and reliable measure of HIV self-management in this population. Differences in aspects of self-management may be related to social roles and community resources, and interventions targeting these factors may decrease morbidity in WLHA.

  16. Performance evaluation of the Bio-Rad Laboratories GS HIV Combo Ag/Ab EIA, a 4th generation HIV assay for the simultaneous detection of HIV p24 antigen and antibodies to HIV-1 (groups M and O) and HIV-2 in human serum or plasma.

    Science.gov (United States)

    Bentsen, Christopher; McLaughlin, Lisa; Mitchell, Elizabeth; Ferrera, Carol; Liska, Sally; Myers, Robert; Peel, Sheila; Swenson, Paul; Gadelle, Stephane; Shriver, M Kathleen

    2011-12-01

    A multi-center study was conducted to evaluate the Bio-Rad GS HIV Combo Ag/Ab EIA, a 4th generation HIV-1/HIV-2 assay for the simultaneous detection of HIV p24 antigen and antibodies to HIV-1 (groups M and O) and HIV-2 in human serum or plasma in adult and pediatric populations. The objectives of the study were to assess assay performance for the detection of acute HIV infections; sensitivity in known HIV positive samples; percent agreement with HIV status; specificity in low and high risk individuals of unknown HIV status; and to compare assay performance to a 3rd generation HIV assay. The evaluation included testing 9150 samples at four U.S. clinical trial sites, using three kit lots. Unlinked samples were from routine testing, repositories or purchased from vendors. GS HIV Combo Ag/Ab EIA detection in samples from individuals in two separate populations with acute HIV infection was 95.2% (20/21) and 86.4% (38/44). Sensitivity was 100% (1603/1603) in known antibody positive [HIV-1 Groups M and O, and HIV-2] samples. HIV p24 antigen detection was 100% (53/53) in HIV-1 culture supernatants. HIV-1 seroconversion panel detection improved by a range of 0-20 days compared to a 3rd generation HIV test. Specificity was 99.9% (5989/5996) in low risk, 99.9% (959/960) in high risk and 100% (100/100) in pediatric populations. The GS HIV Combo Ag/Ab EIA significantly reduced the diagnostic window when compared to the 3rd generation screening assay, enabling earlier diagnosis of HIV infection. The performance parameters of the Bio-Rad GS HIV Combo Ag/Ab EIA are well suited for use in HIV diagnostic settings. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Critical Contribution of Tyr15 in the HIV-1 Integrase (IN) in Facilitating IN Assembly and Nonenzymatic Function through the IN Precursor Form with Reverse Transcriptase.

    Science.gov (United States)

    Takahata, Tatsuro; Takeda, Eri; Tobiume, Minoru; Tokunaga, Kenzo; Yokoyama, Masaru; Huang, Yu-Lun; Hasegawa, Atsuhiko; Shioda, Tatsuo; Sato, Hironori; Kannagi, Mari; Masuda, Takao

    2017-01-01

    Nonenzymatic roles for HIV-1 integrase (IN) at steps prior to the enzymatic integration step have been reported. To obtain structural and functional insights into the nonenzymatic roles of IN, we performed genetic analyses of HIV-1 IN, focusing on a highly conserved Tyr15 in the N-terminal domain (NTD), which has previously been shown to regulate an equilibrium state between two NTD dimer conformations. Replacement of Tyr15 with alanine, histidine, or tryptophan prevented HIV-1 infection and caused severe impairment of reverse transcription without apparent defects in reverse transcriptase (RT) or in capsid disassembly kinetics after entry into cells. Cross-link analyses of recombinant IN proteins demonstrated that lethal mutations of Tyr15 severely impaired IN structure for assembly. Notably, replacement of Tyr15 with phenylalanine was tolerated for all IN functions, demonstrating that a benzene ring of the aromatic side chain is a key moiety for IN assembly and functions. Additional mutagenic analyses based on previously proposed tetramer models for IN assembly suggested a key role of Tyr15 in facilitating the hydrophobic interaction among IN subunits, together with other proximal residues within the subunit interface. A rescue experiment with a mutated HIV-1 with RT and IN deleted (ΔRT ΔIN) and IN and RT supplied in trans revealed that the nonenzymatic IN function might be exerted through the IN precursor conjugated with RT (RT-IN). Importantly, the lethal mutations of Tyr15 significantly reduced the RT-IN function and assembly. Taken together, Tyr15 seems to play a key role in facilitating the proper assembly of IN and RT on viral RNA through the RT-IN precursor form. Inhibitors of the IN enzymatic strand transfer function (INSTI) have been applied in combination antiretroviral therapies to treat HIV-1-infected patients. Recently, allosteric IN inhibitors (ALLINIs) that interact with HIV-1 IN residues, the locations of which are distinct from the catalytic

  18. Multicenter evaluation of a new 4th generation HIV screening assay Elecsys HIV combi.

    Science.gov (United States)

    Weber, B; Orazi, B; Raineri, A; Thorstensson, R; Bürgisser, P; Mühlbacher, A; Areal, C; Eiras, A; Villaescusa, R; Camacho, R; Diogo, I; Roth, H J; Zahn, I; Bartel, J; Bossi, V; Piro, F; Atamasirikul, K; Permpikul, P; Webber, L; Singh, S

    2006-01-01

    Fourth-generation screening assays which permit a simultaneous detection of human immunodeficiency virus (HIV) antigen and antibody reduce the diagnostic window on average by four days in comparison to third-generation antibody assays. Recently, the new automated Elecsys HIV combi was compared in a multicenter study to alternative fourth- and third-generation assays, p24 antigen test and HIV-1 RNA RT-PCR. A total of 104 serocon-version panels, samples of the acute phase of infection after seroconversion (n = 33), anti-HIV-1 positive specimens (n = 572) from patients in different stages of the disease, 535 subtyped samples from different geographical locations, including group M (subtypes A-J) and group O, anti-HIV-2 positive sera (n = 364), dilutions of cell culture supernatants (n = 60) infected with different HIV-1 subtypes, selected performance panels, 8406 unselected samples from blood donors originating from different blood transfusion centers, 3810 unselected sera from daily routine and from hospitalized patients, 9927 unselected samples from South Africa and 1943 potentially interfering samples were tested with the Elecsys HIV combi. Elecsys HIV combi showed a comparable sensitivity to HIV-1 Ag stand-alone assays for early detection of HIV infection in seroconversion panels. The mean time delay of Elecsys HIV combi (last negative sample + 1 day) in comparison to HIV-1 RT-PCR for 92 panels tested with both methods was 3.23 days. The diagnostic window was reduced with Elecsys HIV combi between 1.56 and 5.32 days in comparison to third-generation assays. The specificity of Elecsys HIV combi in blood donors was 99.80% after repeated testing. Our results show that a fourth-generation assay with improved specificity and sensitivity like the Elecsys HIV combi is suitable for blood donor screening due to its low number of false positives and since it detects HIV p24 antigen with a comparable sensitivity to single antigen assays.

  19. Parallel screening of drug-like natural compounds using Caco-2 cell permeability QSAR model with applicability domain, lipophilic ligand efficiency index and shape property: A case study of HIV-1 reverse transcriptase inhibitors

    Science.gov (United States)

    Patel, Rikin D.; Kumar, Sivakumar Prasanth; Patel, Chirag N.; Shankar, Shetty Shilpa; Pandya, Himanshu A.; Solanki, Hitesh A.

    2017-10-01

    The traditional drug design strategy centrally focuses on optimizing binding affinity with the receptor target and evaluates pharmacokinetic properties at a later stage which causes high rate of attrition in clinical trials. Alternatively, parallel screening allows evaluation of these properties and affinity simultaneously. In a case study to identify leads from natural compounds with experimental HIV-1 reverse transcriptase (RT) inhibition, we integrated various computational approaches including Caco-2 cell permeability QSAR model with applicability domain (AD) to recognize drug-like natural compounds, molecular docking to study HIV-1 RT interactions and shape similarity analysis with known crystal inhibitors having characteristic butterfly-like model. Further, the lipophilic properties of the compounds refined from the process with best scores were examined using lipophilic ligand efficiency (LLE) index. Seven natural compound hits viz. baicalien, (+)-calanolide A, mniopetal F, fagaronine chloride, 3,5,8-trihydroxy-4-quinolone methyl ether derivative, nitidine chloride and palmatine, were prioritized based on LLE score which demonstrated Caco-2 well absorption labeling, encompassment in AD structural coverage, better receptor affinity, shape adaptation and permissible AlogP value. We showed that this integrative approach is successful in lead exploration of natural compounds targeted against HIV-1 RT enzyme.

  20. Evaluation of a rapid and simple fourth-generation HIV screening assay for qualitative detection of HIV p24 antigen and/or antibodies to HIV-1 and HIV-2.

    Science.gov (United States)

    Beelaert, G; Fransen, K

    2010-09-01

    The performance was assessed of a new, rapid, visual and qualitative immunoassay for the detection of HIV p24 antigen (Ag) and antibodies (Ab) to HIV-1 and HIV-2. Characterised serum or plasma specimens from patients diagnosed with HIV infection were tested: 179 samples of known Ab-positive patients harbouring different subtypes of HIV-1 (n=154) and HIV-2 (n=25) and 200 samples from individuals not infected with HIV. The assay's Ag sensitivity was assessed by testing HIV seroconversion panels (n=10) and primary HIV infection specimens (n=57). In addition, the influence of the genetic variability of HIV-1 on Ag detection was evaluated using dilutions of culture supernatants infected with different subtypes (n=50). The performance of the rapid test was compared to a "gold standard" testing algorithm with the use of a single Ag ELISA and with the Vironostika((R)) HIV Uni-Form II Ag/Ab test, a fourth-generation ELISA. The new assay, the Determine HIV-1/2 Combo demonstrated 100% (98.2-100.0) Ab specificity (200/200) and 100% (98.0-100.0) Ab sensitivity (179/179). In these samples, the observed Ag sensitivity was 86.6% (58/67) with the Determine HIV-1/2 Combo test and 92.5% (62/67) with the Vironostika compared to the reference single Ag ELISA. The assay could not detect Ag in one group O, one subtype F and two subtype H cell supernatant isolates. None of the HIV-2 Ag could be detected. Copyright 2010 Elsevier B.V. All rights reserved.

  1. Lack of association between plasma levels of non-nucleoside reverse transcriptase inhibitors & virological outcomes during rifampicin co-administration in HIV-infected TB patients.

    Science.gov (United States)

    Ramachandran, Geetha; Kumar, A K Hemanth; Ponnuraja, C; Ramesh, K; Rajesh, Lakshmi; Chandrasekharan, C; Swaminathan, Soumya

    2013-12-01

    Among patients with HIV-associated tuberculosis (TB), reduced plasma non-nucleoside reverse transcriptase inhibitors (NNRTI) concentrations during rifampicin (RMP) co-administration could lead to HIV treatment failure. This study was undertaken to examine the association between plasma nevirapine (NVP) and efavirenz (EFV) concentrations and virological outcomes in patients infected with HIV-1 and TB. This was a nested study undertaken in a clinical trial of patients with HIV-1 and TB, randomized to two different once-daily antiretroviral treatment (ART) regimens along with anti-TB treatment (ATT). Trough concentrations of plasma NVP and EFV were estimated at months 1 (during ATT and ART) and 6 months (ART only) by HPLC. Plasma HIV-1 RNA level >400 copies/ml or death within 6 months of ART were considered as unfavourable outcomes. Genotyping of CYP2B6 516G>T polymorphism was performed. Twenty nine per cent of patients in NVP arm had an unfavourable outcome at 6 months compared to 9 per cent in EFV arm (PLogistic regression analysis showed CYP2B6 516G>T polymorphism significantly associated with virologic outcome in patients receiving EFV-based regimen. Trough plasma concentrations of NVP and EFV did not show any association with response to ART in patients on ATT and once-daily ART. CYP2B6 516G>T polymorphism was associated with virologic outcome among patients on EFV.

  2. Structure of a Dihydroxycoumarin Active-Site Inhibitor in Complex with the RNase H Domain of HIV-1 Reverse Transcriptase and Structure-Activity Analysis of Inhibitor Analogs

    Science.gov (United States)

    Himmel, Daniel M.; Myshakina, Nataliya S.; Ilina, Tatiana; Van Ry, Alexander; Ho, William C.; Parniak, Michael A.; Arnold, Eddy

    2014-01-01

    HIV encodes four essential enzymes: protease, integrase, reverse transcriptase (RT) associated DNA polymerase, and RT-associated ribonuclease H (RNase H). Current clinically approved anti-AIDS drugs target all HIV enzymatic activities except RNase H, which has proven to be a very difficult target for HIV drug discovery. Our high-throughput screening activities identified the dihydroxycoumarin compound F3284-8495 as a specific inhibitor of RT RNase H, with low micromolar potency in vitro. Optimization of inhibitory potency can be facilitated by structural information about inhibitor-target binding. Here, we report the crystal structure of F3284-8495 bound to the active site of an isolated RNase H domain of HIV-1 RT at a resolution limit of 1.71 Å. From predictions based on this structure, compounds were obtained that showed improved inhibitory activity. Computational analysis suggested structural alterations that could provide additional interactions with RT and thus improve inhibitory potency. These studies established proof-of-concept that F3284-8495 could be used as a favorable chemical scaffold for development of HIV RNase H inhibitors. PMID:24840303

  3. Evaluation of an HIV/AIDS peer education programme South African ...

    African Journals Online (AJOL)

    Evaluation of an HIV/AIDS peer education programme. South African workplace . tn a. Nicola M Sloan, Jonathan E Myers. Objectives. To evaluate a South African workplace HIV I AIDS peer-education programme running since 1997. Methods. In 2001 a cross-sectional study was done of 900 retail-section employees in ...

  4. Evaluation of the combination effect of different antiviral compounds against HIV in vitro

    DEFF Research Database (Denmark)

    Sørensen, A M; Nielsen, C; Mathiesen, Lars Reinhardt

    1993-01-01

    3'-azido-3'deoxythymidine (AZT), a clinically used anti-HIV compound, was evaluated for antiviral effect on HIV infection in combination with other antiviral compounds in vitro. Interactions were evaluated by the median-effect principle and the isobologram technique. Synergistic effect was obtained...... emphasis on nucleoside analogues and compounds influencing the infectivity of the virus....

  5. Evaluating the Measurement Structure of the Abbreviated HIV Stigma Scale in a Sample of African Americans Living with HIV/AIDS

    Science.gov (United States)

    Johnson, Eboneé T.; Yaghmaian, Rana A.; Best, Andrew; Chan, Fong; Burrell, Reginald, Jr.

    2016-01-01

    Purpose: The purpose of this study was to validate the 10-item version of the HIV Stigma Scale (HSS-10) in a sample of African Americans with HIV/AIDS. Method: One hundred and ten African Americans living with HIV/AIDS were recruited from 3 case management agencies in Baton Rouge, Louisiana. Measurement structure of the HSS-10 was evaluated using…

  6. Recommendations for evaluation and management of bone disease in HIV

    National Research Council Canada - National Science Library

    Brown, Todd T; Hoy, Jennifer; Borderi, Marco; Guaraldi, Giovanni; Renjifo, Boris; Vescini, Fabio; Yin, Michael T; Powderly, William G

    2015-01-01

    Thirty-four human immunodeficiency virus (HIV) specialists from 16 countries contributed to this project, whose primary aim was to provide guidance on the screening, diagnosis, and monitoring of bone disease in HIV-infected patients...

  7. Evaluation of some cellular immune index in HIV infected participants

    African Journals Online (AJOL)

    Stage 1). Similarly, 40 HIV seronegative participants served as Control. Blood samples collected from the participants were used for HIV screening and confirmation, CD4+ T cell count, absolute lymphocyte count and percent lymphocyte ...

  8. Cupping reversal in pediatric glaucoma--evaluation of the retinal nerve fiber layer and visual field.

    Science.gov (United States)

    Ely, Amanda L; El-Dairi, Mays A; Freedman, Sharon F

    2014-11-01

    To identify optic nerve head (ONH) cupping reversal and associated optical coherence tomography (OCT) and Humphrey visual field changes in pediatric glaucoma. Retrospective observational case series. Sequential surgical cases of juvenile open-angle glaucoma (OAG) or primary congenital glaucoma (PCG) with sustained postoperative intraocular pressure (IOP) reduction. Group 1 had preoperative and postoperative ONH photographs and OCT; Group 2 had preoperative clinical ONH assessment and postoperative imaging. Cupping evaluation was confirmed by masked glaucoma and neuro-ophthalmology specialists. Of 80 cases, 9 eyes (9 children) met criteria for Group 1; 24 eyes (19 children) met criteria for Group 2. Group 1: Five of 9 eyes (56%) demonstrated cupping reversal, with preoperative vs postoperative mean IOP 34.2 ± 6.6 mm Hg vs 10.6 ± 4.1 mm Hg (P cupping reversal, with preoperative vs postoperative mean IOP 36.1 ± 8.9 mm Hg vs 13.3 ± 2.1 mm Hg (P cup-to-disc ratio. Limitations include small numbers, few reliable Humphrey visual fields, and absent preoperative imaging (Group 2). Some eyes with IOP reduction and ONH cupping reversal show continued RNFL thinning postoperatively. The preoperative ONH cup-to-disc ratio predicted the postoperative RNFL better than the postoperative "reversed and smaller" cup-to-disc ratio. Cupping reversal in pediatric glaucoma may not predict improved ONH health and deserves further study. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. In vitro and ex vivo evaluations on transdermal delivery of the HIV inhibitor IQP-0410.

    Directory of Open Access Journals (Sweden)

    Anthony S Ham

    Full Text Available The aim of this study was to investigate the physicochemical and in vitro/ex vivo characteristics of the pyrmidinedione IQP-0410 formulated into transdermal films. IQP-0410 is a potent therapeutic anti-HIV nonnucleoside reverse transcriptase inhibitor that would be subjected to extensive first pass metabolism, through conventional oral administration. Therefore, IQP-0410 was formulated into ethyl cellulose/HPMC-based transdermal films via solvent casting. In mano evaluations were performed to evaluate gross physical characteristics. In vitro release studies were performed in both Franz cells and USP-4 dissolution vessels. Ex vivo release and permeability assays were performed on human epidermal tissue models, and the permeated IQP-0410 was collected for in vitro HIV-1 efficacy assays in CEM-SS cells and PBMCs. Film formulation D3 resulted in pliable, strong transdermal films that were loaded with 2% (w/w IQP-0410. Composed of 60% (w/w ethyl cellulose and 20% (w/w HPMC, the films contained < 1.2% (w/w of water and were hygroscopic resulting in significant swelling under humid conditions. The water permeable nature of the film resulted in complete in vitro dissolution and drug release in 26 hours. When applied to ex vivo epidermal tissues, the films were non-toxic to the tissue and also were non-toxic to HIV target cells used in the in vitro efficacy assays. Over a 3 day application, the films delivered IQP-0410 through the skin tissue at a zero-order rate of 0.94 ± 0.06 µg/cm(2/hr with 134 ± 14.7 µM collected in the basal media. The delivered IQP-0410 resulted in in vitro EC50 values against HIV-1 of 2.56 ± 0.40 nM (CEM-SS and 0.58 ± 0.03 nM (PBMC. The film formulation demonstrated no significant deviation from target values when packaged in foil pouches under standard and accelerated environmental conditions. It was concluded that the transdermal film formulation was a potentially viable method of administering IQP-0410 that warrants

  10. Evaluation of HIV testing recommendations in specialty guidelines for the management of HIV indicator conditions

    DEFF Research Database (Denmark)

    Lord, E; Stockdale, A J; Malek, R

    2017-01-01

    OBJECTIVES: European guidelines recommend HIV testing for individuals presenting with indicator conditions (ICs) including AIDS-defining conditions (ADCs). The extent to which non-HIV specialty guidelines recommend HIV testing in ICs and ADCs is unknown. Our aim was to pilot a methodology in the UK...... to review specialty guidelines and ascertain if HIV was discussed and testing recommended. METHODS: UK and European HIV testing guidelines were reviewed to produce a list of 25 ADCs and 49 ICs. UK guidelines for these conditions were identified from searches of the websites of specialist societies...... (48%) and 36 of 49 (73%) ICs. In total, 78 guidelines were reviewed (range 0-13 per condition). HIV testing was recommended in six of 17 ADC guidelines (35%) and 24 of 61 IC guidelines (39%). At least one guideline recommended HIV testing for six of 25 ADCs (24%) and 16 of 49 ICs (33...

  11. Multicenter study of skin rashes and hepatotoxicity in antiretroviral-na?ve HIV-positive patients receiving non-nucleoside reverse-transcriptase inhibitor plus nucleoside reverse-transcriptase inhibitors in Taiwan

    OpenAIRE

    Wu, Pei-Ying; Cheng, Chien-Yu; Liu, Chun-Eng; Lee, Yi-Chien; Yang, Chia-Jui; Tsai, Mao-Song; Cheng, Shu-Hsing; Lin, Shih-Ping; Lin, De-Yu; Wang, Ning-Chi; Lee, Yi-Chieh; Sun, Hsin-Yun; Tang, Hung-Jen; Hung, Chien-Ching

    2017-01-01

    Objectives Two nucleos(t)ide reverse-transcriptase inhibitors (NRTIs) plus 1 non-NRTI (nNRTI) remain the preferred or alternative combination antiretroviral therapy (cART) for antiretroviral-naive HIV-positive patients in Taiwan. The three most commonly used nNRTIs are nevirapine (NVP), efavirenz (EFV) and rilpivirine (RPV). This study aimed to determine the incidences of hepatotoxicity and skin rashes within 4 weeks of initiation of cART containing 1 nNRTI plus 2 NRTIs. Methods Between June,...

  12. Reverse Transcriptase drug resistance mutations in HIV-1 Subtype C infected patients on ART in Karonga District, Malawi

    LENUS (Irish Health Repository)

    Bansode, Vijay B

    2011-10-13

    Abstract Background Drug resistance testing before initiation of, or during, antiretroviral therapy (ART) is not routinely performed in resource-limited settings. High levels of viral resistance circulating within the population will have impact on treatment programs by increasing the chances of transmission of resistant strains and treatment failure. Here, we investigate Drug Resistance Mutations (DRMs) from blood samples obtained at regular intervals from patients on ART (Baseline-22 months) in Karonga District, Malawi. One hundred and forty nine reverse transcriptase (RT) consensus sequences were obtained via nested PCR and automated sequencing from blood samples collected at three-month intervals from 75 HIV-1 subtype C infected individuals in the ART programme. Results Fifteen individuals showed DRMs, and in ten individuals DRMs were seen from baseline samples (reported to be ART naïve). Three individuals in whom no DRMs were observed at baseline showed the emergence of DRMs during ART exposure. Four individuals who did show DRMs at baseline showed additional DRMs at subsequent time points, while two individuals showed evidence of DRMs at baseline and either no DRMs, or different DRMs, at later timepoints. Three individuals had immune failure but none appeared to be failing clinically. Conclusion Despite the presence of DRMs to drugs included in the current regimen in some individuals, and immune failure in three, no signs of clinical failure were seen during this study. This cohort will continue to be monitored as part of the Karonga Prevention Study so that the long-term impact of these mutations can be assessed. Documenting proviral population is also important in monitoring the emergence of drug resistance as selective pressure provided by ART compromises the current plasma population, archived viruses can re-emerge

  13. Brazilian Program For HIV Point-Of-Care Test Evaluation: A Decade’s Experience

    Directory of Open Access Journals (Sweden)

    Orlando da Costa Ferreira Jr.

    2017-11-01

    Full Text Available The point-of-care tests (POCTs for HIV diagnosis have been widely used in Brazil in order to expand and to allow HIV diagnosis outside health units including remote areas, such as the Amazon region. In order to guarantee the quality of HIV diagnostics based on rapid tests, the Brazilian Ministry of Health (MoH implemented the HIV POCT Evaluation Program. This study compiles the Brazilian experience acquired over the last 13 years conducting the HIV POCT Evaluation Program.   Methods and Findings The selection of tests was based on the interest of manufacturers to qualify for the MoH tenders. Each round was performed with fresh whole blood and oral fluid samples, always including HIV positive and negative ones. In addition to the POCT, every sample was submitted to a reference testing protocol, based on an immunoassay followed by Western blot. The POCTs were evaluated for clinical sensitivity, clinical specificity, assay operational characteristics, detection of HIV-2 antibodies, sensitivity to subtypes panels; and sensitivity to seroconversion panels. Since its implementation in 2003, the POCT evaluation protocol has undergone some modifications aiming to improve and simplify the evaluation process, to know: (i  for HIV-positive samples, perform EIA and Western blot only if the POCT is non-reactive; (ii reduction from 800 to 600 HIV negative samples; (iii increase from one to three subtype panels (including HIV-2 samples; and (iv inclusion of seroconversion panel. We evaluated six tests, four of which met the sensitivity criteria of 99.5%: BD Chek™ HIV Multi-test (whole blood, HIV 1/2 Colloidal Gold (whole blood, OraQuick ADVANCE® Rapid HIV-1/2 Antibody Test (whole blood and oral fluid and TR DPP HIV-1/2 (whole blood, plasma and oral fluid. Regarding other evaluated criteria, all assays met the requirements.   Conclusions The successful Brazilian policy on POCT use for HIV infection diagnosis includes the evaluation of the POCT itself in

  14. Salinity-gradient power: Evaluation of pressure-retarded osmosis and reverse electrodialysis

    NARCIS (Netherlands)

    Post, Jan W.; Veerman, J.A.; Hamelers, Hubertus V.M.; Euverink, Gerrit J.W.; Metz, S.J.; Nijmeijer, Dorothea C.; Buisman, Cees J.N.

    2007-01-01

    A huge potential to obtain clean energy exists from mixing water streams with different salt concentrations. Two membrane-based energy conversion techniques are evaluated: pressure-retarded osmosis and reverse electrodialysis. From the literature, a comparison is not possible since the reported

  15. Salinity-gradient power : Evaluation of pressure-retarded osmosis and reverse electrodialysis

    NARCIS (Netherlands)

    Post, Jan W.; Veerman, Joost; Hamelers, Hubertus V.M.; Euverink, Gerrit J.W.; Metz, Sybrand J.; Nymeijer, Kitty; Buisman, Cees J.N.

    2007-01-01

    A huge potential to obtain clean energy exists from mixing water streams with different salt concentrations. Two membrane-based energy conversion techniques are evaluated: pressure-retarded osmosis and reverse electrodialysis. From the literature, a comparison is not possible since the reported

  16. Visceral Leishmaniasis/HIV co-infection in northeast Brazil: evaluation of outcome

    OpenAIRE

    Távora,Lara Gurgel Fernandes; Nogueira, Marina Bizerril; Gomes, Sofia Teixeira

    2015-01-01

    ABSTRACT Since the beginning of the HIV burden, Visceral Leishmaniasis (VL)/HIV co-infection has been diagnosed not only in areas where VL is endemic (Latin America, India, Asia, Southern Europe), but also in North America, were it is considered an opportunistic disease. Clinical presentation, diagnostic tests sensitivity and treatment response in this population differs from VL alone. OBJECTIVES: To evaluate factors related to an unfavorable outcome in patients with VL/HIV diagnosis in a r...

  17. Evaluation of HIV testing recommendations in specialty guidelines for the management of HIV indicator conditions.

    Science.gov (United States)

    Lord, E; Stockdale, A J; Malek, R; Rae, C; Sperle, I; Raben, D; Freedman, A; Churchill, D; Lundgren, J; Sullivan, A K

    2017-04-01

    European guidelines recommend HIV testing for individuals presenting with indicator conditions (ICs) including AIDS-defining conditions (ADCs). The extent to which non-HIV specialty guidelines recommend HIV testing in ICs and ADCs is unknown. Our aim was to pilot a methodology in the UK to review specialty guidelines and ascertain if HIV was discussed and testing recommended. UK and European HIV testing guidelines were reviewed to produce a list of 25 ADCs and 49 ICs. UK guidelines for these conditions were identified from searches of the websites of specialist societies, the National Institute of Clinical Excellence (NICE) website, the NICE Clinical Knowledge Summaries (CKS) website, the Scottish Intercollegiate Guidance Network (SIGN) website and the British Medical Journal Best Practice database and from Google searches. We identified guidelines for 12 of 25 ADCs (48%) and 36 of 49 (73%) ICs. In total, 78 guidelines were reviewed (range 0-13 per condition). HIV testing was recommended in six of 17 ADC guidelines (35%) and 24 of 61 IC guidelines (39%). At least one guideline recommended HIV testing for six of 25 ADCs (24%) and 16 of 49 ICs (33%). There was no association between recommendation to test and publication year (P = 0.62). The majority of guidelines for ICs do not recommend testing. Clinicians managing ICs may be unaware of recommendations produced by HIV societies or the prevalence of undiagnosed HIV infection among these patients. We are piloting methods to engage with guideline development groups to ensure that patients diagnosed with ICs/ADCs are tested for HIV. We then plan to apply our methodology in other European settings as part of the Optimising Testing and Linkage to Care for HIV across Europe (OptTEST) project. © 2016 The Authors. HIV Medicine published by John Wiley & Sons Ltd on behalf of British HIV Association.

  18. HIV-1 transmission during early antiretroviral therapy: evaluation of two HIV-1 transmission events in the HPTN 052 prevention study.

    Directory of Open Access Journals (Sweden)

    Li-Hua Ping

    Full Text Available In the HPTN 052 study, transmission between HIV-discordant couples was reduced by 96% when the HIV-infected partner received suppressive antiretroviral therapy (ART. We examined two transmission events where the newly infected partner was diagnosed after the HIV-infected partner (index initiated therapy. We evaluated the sequence complexity of the viral populations and antibody reactivity in the newly infected partner to estimate the dates of transmission to the newly infected partners. In both cases, transmission most likely occurred significantly before HIV-1 diagnosis of the newly infected partner, and either just before the initiation of therapy or before viral replication was adequately suppressed by therapy of the index. This study further strengthens the conclusion about the efficacy of blocking transmission by treating the infected partner of discordant couples. However, this study does not rule out the potential for HIV-1 transmission to occur shortly after initiation of ART, and this should be recognized when antiretroviral therapy is used for HIV-1 prevention.

  19. Body image in women with HIV: a cross-sectional evaluation

    Directory of Open Access Journals (Sweden)

    Becerra Karen

    2006-07-01

    Full Text Available Abstract Background HIV lipodystrophy syndrome is a recognized complication of potent antiretroviral therapy and is characterized by often dramatic changes in various body fat stores, both central and peripheral. Given prior findings of heightened body image dysphoria among HIV-infected men with lipodystrophy as compared to HIV-infected men without lipodystrophy, we sought to determine body image among HIV-infected and HIV-negative women and to determine the relationship of HIV and lipodystrophy with body image. Our a priori hypothesis was that women with HIV and lipodystrophy would have significantly poorer body image as compared to women without HIV and to women with HIV without lipodystrophy. Results 116 women responded to two previously validated self-report instruments (Body Image Quality of Life Index (BIQLI and the Situational Inventory of Body-Image Dysphoria – Short Form (SIBID-S on body image. 62 (53% subjects HIV-infected women were recruited at the university-based HIV clinic. 54 (47% subjects HIV-negative female controls were recruited from another study evaluating bone density in otherwise healthy controls. 96% identified their sexual orientation as women having sex with men. Among the HIV-infected group, 36 reported the presence of lipodystrophic characteristics and 26 reported no lipodystrophic changes. Agreement regarding the presence of lipodystrophy between physician and subject was 0.67 as measured by the kappa coefficient of agreement. Compared to HIV-negative women, HIV-positive women demonstrated poor body image as measured by BIQLI (p = 0.0009. Compared with HIV-infected women who denied lipodystrophy, HIV-infected women with self-reported lipodystrophy demonstrated poor body image as measured by BIQLI (p = 0.02 and SIBID-S scales (p = 0.001. Conclusion We demonstrate that HIV and lipodystrophy status among women is associated with poor body image. Universal efforts should be made in the HIV medical community to

  20. Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs.

    Science.gov (United States)

    Parker, W B; White, E L; Shaddix, S C; Ross, L J; Buckheit, R W; Germany, J M; Secrist, J A; Vince, R; Shannon, W M

    1991-01-25

    Carbovir (the carbocyclic analog of 2'-3'-didehydro-2',3'-dideoxyguanosine) is a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication. Assays were developed to assess the mechanism of inhibition by the 5'-triphosphate of carbovir of HIV-1 reverse transcriptase using either RNA or DNA templates that contain all four natural nucleotides. Carbovir-TP was a potent inhibitor of HIV-1 reverse transcriptase using either template with Ki values similar to that observed by AZT-TP, ddGTP, and ddTTP. The kinetic constants for incorporation of these nucleotide analogs into DNA by HIV-1 reverse transcriptase using either template were similar to the values seen for their respective natural nucleotides. In addition, the incorporation of either carbovir-TP or AZT-TP in the presence of dGTP or dTTP, respectively, indicated that the mechanism of inhibition by these two nucleotide analogs was due to their incorporation into the DNA resulting in chain termination. Carbovir-TP was not a potent inhibitor of DNA polymerase alpha, beta, or gamma, or DNA primase. Given the potent activity of carbovir-TP against HIV-1 reverse transcriptase and its lack of activity against human DNA polymerases, we believe that further evaluation of this compound as a potential drug for the treatment of HIV-1 infection is warranted.

  1. Performance evaluation of the point-of-care INSTI™ HIV-1/2 antibody test in early and established HIV infections.

    Science.gov (United States)

    Adams, Sarah; Luo, Wei; Wesolowski, Laura; Cohen, Stephanie E; Peters, Philip J; Owen, S Michele; Masciotra, Silvina

    2017-06-01

    The flow-through INSTI™ HIV-1/HIV-2 Rapid Antibody (INSTI) test is a 60s FDA-approved test for HIV-1 and HIV-2 antibody testing using whole blood and plasma. We evaluated the performance of INSTI using plasma and simulated whole blood specimens. INSTI's performance in plasma specimens from commercial seroconversion panels was assessed by estimating the relative sensitivity using a 50% cumulative frequency analysis and by comparing its performance with other FDA-approved rapid tests (RTs). INSTI was further evaluated using 320 HIV-1 plasma specimens collected during a cross-sectional study and with 107 HIV-1 and 24 HIV-2 simulated whole blood specimens. Sensitivity and specificity were calculated using 615 known HIV-1 group M/O and 80 HIV-2 (Western blot (WB)-positive), and 497 HIV-negative plasma specimens, respectively. In HIV-1 seroconversion panels, INSTI became reactive 9days before a positive WB. When compared to FDA-approved antibody-based lateral flow RTs, INSTI detected significantly more early infections. Among HIV-1-infected cross-sectional plasma samples, INSTI detected 23 (27%) of 85 Architect-positive/Multispot-negative or indeterminate specimens. For plasma specimens, the sensitivity was 99.84% for HIV-1 and 100% for HIV-2, and the specificity was 99.80%. Using simulated whole blood from seroconverters, INSTI performed similarly to plasma. INSTI performed significantly better than antibody-based lateral flow RTs during early stages of seroconversion. Sensitivity and specificity were within the manufacturer's reported ranges. Considering the observed test performance and the almost immediate results, INSTI is an accurate option to detect HIV-1/HIV-2 antibodies in point-of-care settings where lab testing is not feasible. Published by Elsevier B.V.

  2. Reação reversa atípica em paciente com hanseníase dimorfa co-infectado pelo HIV Atypical reversal reaction in a borderline leprosy patient co-infected with HIV

    Directory of Open Access Journals (Sweden)

    Rafaela de Lima Caruso

    2007-12-01

    Full Text Available A infecção pelo HIV não altera a história natural da hanseníase. Observa-se maior incidência de estados reacionais nos pacientes co-infectados, além de casos mais graves de neurite. Paciente soropositiva com hanseníase dimorfa tuberculóide manifestou reação reversa exuberante. Lesões cutâneas atípicas e raras surgiram após a introdução da terapia anti-retroviral que promoveu o início da recuperação imunológica com aumento de linfócitos T CD4+ e queda da carga viral. A restauração da imunidade celular nos pacientes soropositivos pode precipitar reações reversas, descritas recentemente como uma das manifestações da síndrome inflamatória de reconstituição imunológica associada à hanseníase.HIV infection does not modify the natural course of leprosy. However, HIV co-infection seems to be associated with an increased rate of reactional states and more severe cases of neuritis. The authors describe a case of borderline tuberculoid leprosy in a HIV-positive patient who developed a marked reversal reaction. Atypical and rare skin manifestations, such as verrucous lesions and ulcers, appeared after highly active antiretroviral therapy, which resulted in increased CD4+ T-lymphocyte count and drop in viral load. The restoration of the cellular immunity in HIV-seropositive patients can trigger reversal reactions that are one of the manifestations of the immune reconstitution inflammatory syndrome. Only recently the syndrome has been associated with leprosy.

  3. Performance evaluation of the ADVIA Centaur(®) HIV Ag/Ab Combo assay.

    Science.gov (United States)

    Pumarola, Tomàs; Freeman, James; Saxton, Emeline; Dillon, Paul; Bal, Tricia; van Helden, Josef

    2010-12-01

    Early diagnosis of HIV infection and appropriate care reduces morbidity and mortality. As a result, recent guidelines recommend that HIV screening be routinely included in patient care. Routine screening will likely result in more patients being tested prior to seroconversion; fourth-generation assays can facilitate diagnosis in these patients. This study evaluated the performance of the automated fourth-generation ADVIA Centaur(®) HIV Ag/Ab Combo assay. Samples from three sites were tested using the HIV Ag/Ab Combo assay and a CE-marked predicate assay. The HIV Ag/Ab Combo assay's relative sensitivity was 98.36% (599/609; 95% confidence interval: 97.00-99.21%), and the relative specificity was 99.74% (7743/7763; 95% confidence interval: 99.60-99.84%). The HIV Ag/Ab Combo assay detected seroconversion at the same bleed or at least one bleed earlier in 34/37 panels compared to the CE-marked predicate assay. Compared to the final result, the HIV Ag/Ab Combo assay's sensitivity was 100% (598/598; 95% confidence interval: 99.39-100.00%), and the specificity was 99.74% (7753/7773; 95% confidence interval: 99.60-99.84%). Sensitivity was 100% for the HIV genotypes tested. The HIV Ag/Ab Combo assay is a sensitive and specific assay that can assist clinicians in the early diagnosis of HIV infection. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Evaluation of Anti-HIV-1 Mutagenic Nucleoside Analogues*

    Science.gov (United States)

    Vivet-Boudou, Valérie; Isel, Catherine; El Safadi, Yazan; Smyth, Redmond P.; Laumond, Géraldine; Moog, Christiane; Paillart, Jean-Christophe; Marquet, Roland

    2015-01-01

    Because of their high mutation rates, RNA viruses and retroviruses replicate close to the threshold of viability. Their existence as quasi-species has pioneered the concept of “lethal mutagenesis” that prompted us to synthesize pyrimidine nucleoside analogues with antiviral activity in cell culture consistent with an accumulation of deleterious mutations in the HIV-1 genome. However, testing all potentially mutagenic compounds in cell-based assays is tedious and costly. Here, we describe two simple in vitro biophysical/biochemical assays that allow prediction of the mutagenic potential of deoxyribonucleoside analogues. The first assay compares the thermal stabilities of matched and mismatched base pairs in DNA duplexes containing or not the nucleoside analogues as follows. A promising candidate should display a small destabilization of the matched base pair compared with the natural nucleoside and the smallest gap possible between the stabilities of the matched and mismatched base pairs. From this assay, we predicted that two of our compounds, 5-hydroxymethyl-2′-deoxyuridine and 5-hydroxymethyl-2′-deoxycytidine, should be mutagenic. The second in vitro reverse transcription assay assesses DNA synthesis opposite nucleoside analogues inserted into a template strand and subsequent extension of the newly synthesized base pairs. Once again, only 5-hydroxymethyl-2′-deoxyuridine and 5-hydroxymethyl-2′-deoxycytidine are predicted to be efficient mutagens. The predictive potential of our fast and easy first line screens was confirmed by detailed analysis of the mutation spectrum induced by the compounds in cell culture because only compounds 5-hydroxymethyl-2′-deoxyuridine and 5-hydroxymethyl-2′-deoxycytidine were found to increase the mutation frequency by 3.1- and 3.4-fold, respectively. PMID:25398876

  5. Evaluation of anti-HIV-1 mutagenic nucleoside analogues.

    Science.gov (United States)

    Vivet-Boudou, Valérie; Isel, Catherine; El Safadi, Yazan; Smyth, Redmond P; Laumond, Géraldine; Moog, Christiane; Paillart, Jean-Christophe; Marquet, Roland

    2015-01-02

    Because of their high mutation rates, RNA viruses and retroviruses replicate close to the threshold of viability. Their existence as quasi-species has pioneered the concept of "lethal mutagenesis" that prompted us to synthesize pyrimidine nucleoside analogues with antiviral activity in cell culture consistent with an accumulation of deleterious mutations in the HIV-1 genome. However, testing all potentially mutagenic compounds in cell-based assays is tedious and costly. Here, we describe two simple in vitro biophysical/biochemical assays that allow prediction of the mutagenic potential of deoxyribonucleoside analogues. The first assay compares the thermal stabilities of matched and mismatched base pairs in DNA duplexes containing or not the nucleoside analogues as follows. A promising candidate should display a small destabilization of the matched base pair compared with the natural nucleoside and the smallest gap possible between the stabilities of the matched and mismatched base pairs. From this assay, we predicted that two of our compounds, 5-hydroxymethyl-2'-deoxyuridine and 5-hydroxymethyl-2'-deoxycytidine, should be mutagenic. The second in vitro reverse transcription assay assesses DNA synthesis opposite nucleoside analogues inserted into a template strand and subsequent extension of the newly synthesized base pairs. Once again, only 5-hydroxymethyl-2'-deoxyuridine and 5-hydroxymethyl-2'-deoxycytidine are predicted to be efficient mutagens. The predictive potential of our fast and easy first line screens was confirmed by detailed analysis of the mutation spectrum induced by the compounds in cell culture because only compounds 5-hydroxymethyl-2'-deoxyuridine and 5-hydroxymethyl-2'-deoxycytidine were found to increase the mutation frequency by 3.1- and 3.4-fold, respectively. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Inhibition of HIV-1 reverse transcriptase-catalyzed synthesis by intercalated DNA Benzo[a]Pyrene 7,8-Dihydrodiol-9,10-Epoxide adducts.

    Directory of Open Access Journals (Sweden)

    Parvathi Chary

    Full Text Available To aid in the characterization of the relationship of structure and function for human immunodeficiency virus type-1 reverse transcriptase (HIV-1 RT, this investigation utilized DNAs containing benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE-modified primers and templates as a probe of the architecture of this complex. BPDE lesions that differed in their stereochemistry around the C10 position were covalently linked to N (6-adenine and positioned in either the primer or template strand of a duplex template-primer. HIV-1 RT exhibited a stereoisomer-specific and strand-specific difference in replication when the BPDE-lesion was placed in the template versus the primer strand. When the C10 R-BPDE adduct was positioned in the primer strand in duplex DNA, 5 nucleotides from the 3΄ end of the primer terminus, HIV-1 RT could not fully replicate the template, producing truncated products; this block to further synthesis did not affect rates of dissociation or DNA binding affinity. Additionally, when the adducts were in the same relative position, but located in the template strand, similar truncated products were observed with both the C10 R and C10 S BPDE adducts. These data suggest that the presence of covalently-linked intercalative DNA adducts distant from the active site can lead to termination of DNA synthesis catalyzed by HIV-1 RT.

  7. An Evaluation of Selected Populations for HIV-1 Vaccine Cohort Development in Nigeria.

    Directory of Open Access Journals (Sweden)

    Ogbonnaya S Njoku

    Full Text Available Development of a globally effective HIV-1 vaccine will need to encompass Nigeria, one of the hardest hit areas, with an estimated 3.2 million people living with HIV. This cross-sectional Institutional Review Board (IRB approved study was conducted in 2009-12 at four market sites and two highway settlements sites in Nigeria to identify and characterize populations at high risk for HIV; engage support of local stakeholders; and assess the level of interest in future vaccine studies. Demographic, HIV risk data were collected by structured interviewer-administered questionnaires. Blood samples were tested on site by HIV rapid diagnostic tests, followed by rigorous confirmatory testing, subtype evaluation and testing for HBV and HCV markers in a clinical reference laboratory. Of 3229 study participants, 326 were HIV infected as confirmed by Western Blot or RNA, with a HIV prevalence of 15.4%-23.9% at highway settlements and 3.1%-9.1% at market sites. There was no observable correlation of prevalence of HIV-1 (10.1% with HBV (10.9% or HCV (2.9%. Major HIV-1 subtypes included CRF02_AG (37.5%; G (27.5%; G/CRF02_AG (25.9%; and non-typeable (8.9%, with 0.3% HIV-2. Univariate analysis found age, gender, marital status, level of education, and sex under substance influence as significant risk factors for HIV (p<0.001. Educating and winning the trust of local community leadership ensured high level of participation (53.3-77.9% and willingness to participate in future studies (95%. The high HIV prevalence and high risk of HIV infection at highway settlement and mammy markets make them well suited for targeting future vaccine trials in Nigeria.

  8. Field evaluation of the Abbott ARCHITECT HIV Ag/Ab Combo immunoassay.

    Science.gov (United States)

    Kfutwah, Anfumbom; Lemée, Véronique; Ngono, Hélène Valérie; De Oliveira, Fabienne; Njouom, Richard; Plantier, Jean-Christophe

    2013-12-01

    Fourth generation assays for HIV diagnosis are progressively being introduced into routine services, due to their improvement of diagnosis. In spite of this, HIV diagnosis remains a challenge in sub-Saharan Africa, due to false positive reactivity. There is a continuous need for field evaluations and routine validations of fourth generation HIV tests in African populations. Evaluate the performances of the ARCHITECT HIV Ag/Ab kit (Abbott) in a population living in an African setting-Cameroon compared to a population living in a European setting-France. 645 HIV samples from both France and Cameroon were evaluated. The positive panel (378 samples) included a diverse series of HIV-1 variants (groups M, N, O, and P) as well as HIV-2 samples. Results were compared to original diagnosis done with other 4th generation assays (AxSYM HIV Ag/Ab (Abbott) and Vidas HIV DUO QUICK) (bioMérieux). Sensitivity of the ARCHITECT was 100% in both sites. It diagnosed all variants of the panel with different reactivity profiles following strain diversity. A wider range of reactivity was observed for group O. Specificity was slightly lower (97.6%) in Cameroon than in France (98.6%), probably due to a higher rate of false positive reactivity. ARCHITECT HIV Ag/Ab assay had high performances in clinical sensitivity and specificity and is adapted to the wide genetic diversity of viruses circulating in West Central Africa. Our results further highlight the need to evaluate HIV diagnostic tests before introduction into routine diagnostic services both in the North and in the South. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Evaluation of accuracy of OraQuick ® rapid test in detecting HIV ...

    African Journals Online (AJOL)

    Objective: The accuracy of OraQuick® rapid test in detecting HIV 1 & 2 antibodies in saliva is evaluated against the blood EIA benchmark tests with confirmatory testing, against which OraQuick® accuracy is determined. Method: Paired samples of saliva and blood from 281 Nigerians were tested for HIV antibodies, and ...

  10. Reporting and evaluation of HIV-related clinical endpoints in two multicenter international clinical trials

    DEFF Research Database (Denmark)

    Lifson, A; Rahme, FS; Belloso, WH

    2006-01-01

    PURPOSE: The processes for reporting and review of progression of HIV disease clinical endpoints are described for two large phase III international clinical trials. METHOD: SILCAAT and ESPRIT are multicenter randomized HIV trials evaluating the impact of interleukin-2 on disease progression...

  11. Monitoring and evaluation of sport-based HIV/AIDS awareness ...

    African Journals Online (AJOL)

    Elma Nelisiwe Maleka

    2016-12-20

    Dec 20, 2016 ... AIDS awareness programmes: Strengthening outcome indicators. Elma Nelisiwe Maleka. To cite this article: Elma Nelisiwe Maleka (2017) Monitoring and evaluation of sport-based HIV/. AIDS awareness programmes: Strengthening outcome indicators, SAHARA-J: Journal of Social. Aspects of HIV/AIDS, ...

  12. Evaluation of a new fourth generation enzyme-linked immunosorbent assay, the LG HIV Ag-Ab Plus, with a combined HIV p24 antigen and anti-HIV-1/2/O screening test.

    Science.gov (United States)

    Yeom, Joon-Sup; Jun, Gyo; Chang, Young; Sohn, Mi-Jin; Yoo, Seungbum; Kim, Eunkyung; Ryu, Seung-Ho; Kang, Hee-Jung; Kim, Young-A; Ahn, Sun-Young; Cha, Je-Eun; Youn, Sung-Tae; Park, Jae-Won

    2006-11-01

    The LG HIV Ag-Ab Plus, a new fourth generation diagnostic assay for HIV infection, was evaluated in comparison to the Enzygnost HIV Integral, an established fourth generation HIV assay. The LG assay showed 100% sensitivity with 109 samples with anti-HIV-1, anti-HIV-2 or anti-HIV-1 group O reactivity. It also detected correctly all 51 positives on three BBI performance panels, slightly outperforming the Enzygnost HIV Integral, which detected 50. The specificity of the LG HIV Ag-Ab Plus was 99.9% with 999 sera from healthy blood donors, which was slightly inferior to the performance of the Enzygnost HIV Integral, which had 100% specificity. The LG assay showed 100% specificity with 81 specimens with underlying diseases including hepatitis B, demonstrating a low risk of cross-reactivity with other infections. The reduction of the diagnostic window by the LG HIV Ag-Ab Plus, compared to a third generation HIV assay, was 6.3 days. The LG assay also showed sufficiently high intra-person and inter-person reproducibility. The overall performance of this new fourth generation HIV assay was adequate for screening and diagnosis of HIV infection.

  13. HIV

    African Journals Online (AJOL)

    Introduction. The·human immunodeficiency virus (HIV) can be transmiHed from one person to onother through the use of non-sterile nee- dles, syringes, and other skin-piercing and invasive instruments. Proper .sterilization of all such instruments is therefore important to prevent its transmission. HIV is very sensitive to ...

  14. A new general method for simultaneous fitting of temperature and concentration dependence of reaction rates yields kinetic and thermodynamic parameters for HIV reverse transcriptase specificity.

    Science.gov (United States)

    Li, An; Ziehr, Jessica L; Johnson, Kenneth A

    2017-04-21

    Recent studies have demonstrated the dominant role of induced fit in enzyme specificity of HIV reverse transcriptase and many other enzymes. However, relevant thermodynamic parameters are lacking, and equilibrium thermodynamic methods are of no avail because the key parameters can only be determined by kinetic measurement. By modifying KinTek Explorer software, we present a new general method for globally fitting data collected over a range of substrate concentrations and temperatures and apply it to HIV reverse transcriptase. Fluorescence stopped-flow methods were used to record the kinetics of enzyme conformational changes that monitor nucleotide binding and incorporation. The nucleotide concentration dependence was measured at temperatures ranging from 5 to 37 °C, and the raw data were fit globally to derive a single set of rate constants at 37 °C and a set of activation enthalpy terms to account for the kinetics at all other temperatures. This comprehensive analysis afforded thermodynamic parameters for nucleotide binding ( K d , Δ G , Δ H , and Δ S at 37 °C) and kinetic parameters for enzyme conformational changes and chemistry (rate constants and activation enthalpy). Comparisons between wild-type enzyme and a mutant resistant to nucleoside analogs used to treat HIV infections reveal that the ground state binding is weaker and the activation enthalpy for the conformational change step is significantly larger for the mutant. Further studies to explore the structural underpinnings of the observed thermodynamics and kinetics of the conformational change step may help to design better analogs to treat HIV infections and other diseases. Our new method is generally applicable to enzyme and chemical kinetics. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Monitoring and evaluation of sport-based HIV/AIDS awareness programmes: Strengthening outcome indicators

    Directory of Open Access Journals (Sweden)

    Elma Nelisiwe Maleka

    2017-01-01

    Full Text Available There are number of Non-Governmental Organisations (NGOs in South Africa that use sport as a tool to respond to Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS, however, little is reported about the outcomes and impact of these programmes. The aim of this study is to contribute to a generic monitoring and evaluation framework by improving the options for the use of outcome indicators of sport-based HIV/AIDS awareness programmes of selected NGOs in South Africa. A qualitative method study was carried out with seven employees of five selected NGOs that integrate sport to deliver HIV/AIDS programmes in South Africa. The study further involved six specialists/experts involved in the field of HIV/AIDS and an official from Sport Recreation South Africa (SRSA. Multiple data collection instruments including desktop review, narrative systematic review, document analysis, one-on-one interviews and focus group interview were used to collect information on outcomes and indicators for sport-based HIV/AIDS awareness programmes. The information was classified according to the determinants of HIV/AIDS. The overall findings revealed that the sport-based HIV/AIDS awareness programmes of five selected NGOs examined in this study focus on similar HIV prevention messages within the key priorities highlighted in the current National Strategic Plan for HIV/AIDS, STIs and TB of South Africa. However, monitoring and evaluating outcomes of sport-based HIV/AIDS programmes of the selected NGOs remains a challenge. A need exists for the improvement of the outcome statements and indicators for their sport-based HIV/AIDS awareness programmes. This study proposed a total of 51 generic outcome indicators focusing on measuring change in the knowledge of HIV/AIDS and change in attitude and intention towards HIV risk behaviours. In addition, this study further proposed a total of eight generic outcome indicators to measure predictors of HIV risk behaviour

  16. Monitoring and evaluation of sport-based HIV/AIDS awareness programmes: Strengthening outcome indicators.

    Science.gov (United States)

    Maleka, Elma Nelisiwe

    2017-12-01

    There are number of Non-Governmental Organisations (NGOs) in South Africa that use sport as a tool to respond to Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS), however, little is reported about the outcomes and impact of these programmes. The aim of this study is to contribute to a generic monitoring and evaluation framework by improving the options for the use of outcome indicators of sport-based HIV/AIDS awareness programmes of selected NGOs in South Africa. A qualitative method study was carried out with seven employees of five selected NGOs that integrate sport to deliver HIV/AIDS programmes in South Africa. The study further involved six specialists/experts involved in the field of HIV/AIDS and an official from Sport Recreation South Africa (SRSA). Multiple data collection instruments including desktop review, narrative systematic review, document analysis, one-on-one interviews and focus group interview were used to collect information on outcomes and indicators for sport-based HIV/AIDS awareness programmes. The information was classified according to the determinants of HIV/AIDS. The overall findings revealed that the sport-based HIV/AIDS awareness programmes of five selected NGOs examined in this study focus on similar HIV prevention messages within the key priorities highlighted in the current National Strategic Plan for HIV/AIDS, STIs and TB of South Africa. However, monitoring and evaluating outcomes of sport-based HIV/AIDS programmes of the selected NGOs remains a challenge. A need exists for the improvement of the outcome statements and indicators for their sport-based HIV/AIDS awareness programmes. This study proposed a total of 51 generic outcome indicators focusing on measuring change in the knowledge of HIV/AIDS and change in attitude and intention towards HIV risk behaviours. In addition, this study further proposed a total of eight generic outcome indicators to measure predictors of HIV risk behaviour. The selected

  17. Monitoring and evaluation of sport-based HIV/AIDS awareness programmes: Strengthening outcome indicators

    Science.gov (United States)

    Maleka, Elma Nelisiwe

    2017-01-01

    Abstract There are number of Non-Governmental Organisations (NGOs) in South Africa that use sport as a tool to respond to Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS), however, little is reported about the outcomes and impact of these programmes. The aim of this study is to contribute to a generic monitoring and evaluation framework by improving the options for the use of outcome indicators of sport-based HIV/AIDS awareness programmes of selected NGOs in South Africa. A qualitative method study was carried out with seven employees of five selected NGOs that integrate sport to deliver HIV/AIDS programmes in South Africa. The study further involved six specialists/experts involved in the field of HIV/AIDS and an official from Sport Recreation South Africa (SRSA). Multiple data collection instruments including desktop review, narrative systematic review, document analysis, one-on-one interviews and focus group interview were used to collect information on outcomes and indicators for sport-based HIV/AIDS awareness programmes. The information was classified according to the determinants of HIV/AIDS. The overall findings revealed that the sport-based HIV/AIDS awareness programmes of five selected NGOs examined in this study focus on similar HIV prevention messages within the key priorities highlighted in the current National Strategic Plan for HIV/AIDS, STIs and TB of South Africa. However, monitoring and evaluating outcomes of sport-based HIV/AIDS programmes of the selected NGOs remains a challenge. A need exists for the improvement of the outcome statements and indicators for their sport-based HIV/AIDS awareness programmes. This study proposed a total of 51 generic outcome indicators focusing on measuring change in the knowledge of HIV/AIDS and change in attitude and intention towards HIV risk behaviours. In addition, this study further proposed a total of eight generic outcome indicators to measure predictors of HIV risk behaviour. The

  18. Ultrasonic guided waves dispersion reversal for long bone thickness evaluation: a simulation study.

    Science.gov (United States)

    Xu, Kailiang; Liu, Chengcheng; Ta, Dean

    2013-01-01

    It has been shown that ultrasonic guided waves have great potentials for long cortical bone evaluation. However, due to the multimodal dispersion, the received signals usually contain several mixed guided modes, which highly complicates the mode separation and signal processing. In the study, we showed that the use of dispersion reversal excitation allows the self-compensation of the dispersive modes in the long cortical bone. Two-dimension finite-difference time-domain (2D-FDTD) method was employed to simulate the propagation of two fundamental guided modes, symmetrical S0 and anti-symmetrical A0, in the long cortical bones. It was demonstrated that the pulse-like modes of S0 and A0 can be detected under the dispersion reversal excitations. The simulations also illustrated that the proposed dispersion reversal method can be used to evaluate the cortical thickness. Results are promising for the application of dispersion reversal method in ultrasonic assessment of the long cortical bone.

  19. Evaluation of simple rapid HIV assays and development of national rapid HIV test algorithms in Dar es Salaam, Tanzania

    Directory of Open Access Journals (Sweden)

    Mbwana Judica

    2009-02-01

    Full Text Available Abstract Background Suitable algorithms based on a combination of two or more simple rapid HIV assays have been shown to have a diagnostic accuracy comparable to double enzyme-linked immunosorbent assay (ELISA or double ELISA with Western Blot strategies. The aims of this study were to evaluate the performance of five simple rapid HIV assays using whole blood samples from HIV-infected patients, pregnant women, voluntary counseling and testing attendees and blood donors, and to formulate an alternative confirmatory strategy based on rapid HIV testing algorithms suitable for use in Tanzania. Methods Five rapid HIV assays: Determine™ HIV-1/2 (Inverness Medical, SD Bioline HIV 1/2 3.0 (Standard Diagnostics Inc., First Response HIV Card 1–2.0 (PMC Medical India Pvt Ltd, HIV1/2 Stat-Pak Dipstick (Chembio Diagnostic System, Inc and Uni-Gold™ HIV-1/2 (Trinity Biotech were evaluated between June and September 2006 using 1433 whole blood samples from hospital patients, pregnant women, voluntary counseling and testing attendees and blood donors. All samples that were reactive on all or any of the five rapid assays and 10% of non-reactive samples were tested on a confirmatory Inno-Lia HIV I/II immunoblot assay (Immunogenetics. Results Three hundred and ninety samples were confirmed HIV-1 antibody positive, while 1043 were HIV negative. The sensitivity at initial testing of Determine, SD Bioline and Uni-Gold™ was 100% (95% CI; 99.1–100 while First Response and Stat-Pak had sensitivity of 99.5% (95% CI; 98.2–99.9 and 97.7% (95% CI; 95.7–98.9, respectively, which increased to 100% (95% CI; 99.1–100 on repeat testing. The initial specificity of the Uni-Gold™ assay was 100% (95% CI; 99.6–100 while specificities were 99.6% (95% CI; 99–99.9, 99.4% (95% CI; 98.8–99.7, 99.6% (95% CI; 99–99.9 and 99.8% (95% CI; 99.3–99.9 for Determine, SD Bioline, First Response and Stat-Pak assays, respectively. There was no any sample which was

  20. Evaluation of the interactions of HIV-1 integrase with small ubiquitin-like modifiers and their conjugation enzyme Ubc9.

    Science.gov (United States)

    Li, Zhihui; Wu, Shuwen; Wang, Jingjing; Li, Wenjuan; Lin, Yun; Ji, Chaoneng; Xue, Jinglun; Chen, Jinzhong

    2012-11-01

    Human immunodeficiency virus type 1 (HIV-1) integrase mediates the integration of reverse-transcribed viral cDNA into the genome of the host for the stable maintenance of the viral genome and the persistence of HIV-1 infection. In this study, the relationships between HIV-1 integrase (HIV-1 IN) and three SUMO conjugation pathway proteins, as well as the effects of these associations, were investigated. The overexpression of SUMO1/SUMO2 and Ubc9 changed the intracellular localization of HIV-1 IN from a diffuse distribution to a punctate localization. SUMO1, SUMO2 and Ubc9 were shown to interact with HIV-1 IN. The SUMOylation of HIV-1 IN was verified. In addition, SUMO1, SUMO2 and Ubc9 were shown to influence the integration of both lentivirus and HIV-1. The overexpression of Ubc9 inhibited viral genome integration, and the upregulation of SUMO1 or SUMO2 enhanced the inhibitory effect of Ubc9. Knockdown of the endogenous levels of SUMO1, SUMO2 and Ubc9 increased the level of viral integration, while reverse transcription and the nuclear import of preintegration complex (PIC) were not affected. Our findings suggest that SUMO conjugation pathway proteins may act as cellular restriction factors and be detrimental to HIV-1 infection. These findings merit further investigation because of their potentially significant implications for the cellular antiviral response to HIV-1 infection.

  1. Selective killing of human immunodeficiency virus infected cells by non-nucleoside reverse transcriptase inhibitor-induced activation of HIV protease

    Directory of Open Access Journals (Sweden)

    Smeulders Liesbeth

    2010-10-01

    Full Text Available Abstract Background Current antiretroviral therapy against human immunodeficiency virus (HIV-1 reduces viral load and thereby prevents viral spread, but it cannot eradicate proviral genomes from infected cells. Cells in immunological sanctuaries as well as cells producing low levels of virus apparently contribute to a reservoir that maintains HIV persistence in the presence of highly active antiretroviral therapy. Thus, accelerated elimination of virus producing cells may represent a complementary strategy to control HIV infection. Here we sought to exploit HIV protease (PR related cytotoxicity in order to develop a strategy for drug induced killing of HIV producing cells. PR processes the viral Gag and Gag-Pol polyproteins during virus maturation, but is also implicated in killing of virus producing cells through off-target cleavage of host proteins. It has been observed previously that micromolar concentrations of certain non-nucleoside reverse transcriptase inhibitors (NNRTIs can stimulate intracellular PR activity, presumably by enhancing Gag-Pol dimerization. Results Using a newly developed cell-based assay we compared the degree of PR activation displayed by various NNRTIs. We identified inhibitors showing higher potency with respect to PR activation than previously described for NNRTIs, with the most potent compounds resulting in ~2-fold increase of the Gag processing signal at 250 nM. The degree of enhancement of intracellular Gag processing correlated with the compound's ability to enhance RT dimerization in a mammalian two-hybrid assay. Compounds were analyzed for their potential to mediate specific killing of chronically infected MT-4 cells. Levels of cytotoxicity on HIV infected cells determined for the different NNRTIs corresponded to the relative degree of drug induced intracellular PR activation, with CC50 values ranging from ~0.3 μM to above the tested concentration range (10 μM. Specific cytotoxicity was reverted by addition

  2. Duplex structural differences and not 2′-hydroxyls explain the more stable binding of HIV-reverse transcriptase to RNA-DNA versus DNA-DNA

    OpenAIRE

    Olimpo, Jeffrey T.; DeStefano, Jeffrey J.

    2010-01-01

    Human immunodeficiency virus reverse transcriptase (HIV-RT) binds more stably in binary complexes with RNA–DNA versus DNA–DNA. Current results indicate that only the -2 and -4 RNA nucleotides (-1 hybridized to the 3′ recessed DNA base) are required for stable binding to RNA–DNA, and even a single RNA nucleotide conferred significantly greater stability than DNA–DNA. Replacing 2′- hydroxyls on pivotal RNA bases with 2′-O-methyls did not affect stability, indicating that interactions between hy...

  3. Multicenter study of skin rashes and hepatotoxicity in antiretroviral-naïve HIV-positive patients receiving non-nucleoside reverse-transcriptase inhibitor plus nucleoside reverse-transcriptase inhibitors in Taiwan.

    Directory of Open Access Journals (Sweden)

    Pei-Ying Wu

    Full Text Available Two nucleos(tide reverse-transcriptase inhibitors (NRTIs plus 1 non-NRTI (nNRTI remain the preferred or alternative combination antiretroviral therapy (cART for antiretroviral-naive HIV-positive patients in Taiwan. The three most commonly used nNRTIs are nevirapine (NVP, efavirenz (EFV and rilpivirine (RPV. This study aimed to determine the incidences of hepatotoxicity and skin rashes within 4 weeks of initiation of cART containing 1 nNRTI plus 2 NRTIs.Between June, 2012 and November, 2015, all antiretroviral-naive HIV-positive adult patients initiating nNRTI-containing cART at 8 designated hospitals for HIV care were included in this retrospective observational study. According to the national HIV treatment guidelines, patients were assessed at baseline, 2 and 4 weeks of cART initiation, and subsequently every 8 to 12 weeks. Plasma HIV RNA load, CD4 cell count and aminotransferases were determined. The toxicity grading scale of the Division of AIDS (DAIDS 2014 was used for reporting clinical and laboratory adverse events.During the 3.5-year study period, 2,341 patients initiated nNRTI-containing cART: NVP in 629 patients, EFV 1,363 patients, and RPV 349 patients. Rash of any grade occurred in 14.1% (n = 331 of the patients. In multiple logistic regression analysis, baseline CD4 cell counts (per 100-cell/μl increase, adjusted odds ratio [AOR], 1.125; 95% confidence interval [95% CI], 1.031-1.228 and use of NVP (AOR, 2.443; 95% CI, 1.816-3.286 (compared with efavirenz were independently associated with the development of skin rashes. Among the 1,455 patients (62.2% with aminotransferase data both at baseline and week 4, 72 (4.9% developed grade 2 or greater hepatotoxicity. In multiple logistic regression analysis, presence of antibody for hepatitis C virus (HCV (AOR, 2.865; 95% CI, 1.439-5.704 or hepatitis B surface antigen (AOR, 2.397; 95% CI, 1.150-4.997, and development of skin rashes (AOR, 2.811; 95% CI, 1.051-7.521 were independently

  4. Multicenter study of skin rashes and hepatotoxicity in antiretroviral-naïve HIV-positive patients receiving non-nucleoside reverse-transcriptase inhibitor plus nucleoside reverse-transcriptase inhibitors in Taiwan.

    Science.gov (United States)

    Wu, Pei-Ying; Cheng, Chien-Yu; Liu, Chun-Eng; Lee, Yi-Chien; Yang, Chia-Jui; Tsai, Mao-Song; Cheng, Shu-Hsing; Lin, Shih-Ping; Lin, De-Yu; Wang, Ning-Chi; Lee, Yi-Chieh; Sun, Hsin-Yun; Tang, Hung-Jen; Hung, Chien-Ching

    2017-01-01

    Two nucleos(t)ide reverse-transcriptase inhibitors (NRTIs) plus 1 non-NRTI (nNRTI) remain the preferred or alternative combination antiretroviral therapy (cART) for antiretroviral-naive HIV-positive patients in Taiwan. The three most commonly used nNRTIs are nevirapine (NVP), efavirenz (EFV) and rilpivirine (RPV). This study aimed to determine the incidences of hepatotoxicity and skin rashes within 4 weeks of initiation of cART containing 1 nNRTI plus 2 NRTIs. Between June, 2012 and November, 2015, all antiretroviral-naive HIV-positive adult patients initiating nNRTI-containing cART at 8 designated hospitals for HIV care were included in this retrospective observational study. According to the national HIV treatment guidelines, patients were assessed at baseline, 2 and 4 weeks of cART initiation, and subsequently every 8 to 12 weeks. Plasma HIV RNA load, CD4 cell count and aminotransferases were determined. The toxicity grading scale of the Division of AIDS (DAIDS) 2014 was used for reporting clinical and laboratory adverse events. During the 3.5-year study period, 2,341 patients initiated nNRTI-containing cART: NVP in 629 patients, EFV 1,363 patients, and RPV 349 patients. Rash of any grade occurred in 14.1% (n = 331) of the patients. In multiple logistic regression analysis, baseline CD4 cell counts (per 100-cell/μl increase, adjusted odds ratio [AOR], 1.125; 95% confidence interval [95% CI], 1.031-1.228) and use of NVP (AOR, 2.443; 95% CI, 1.816-3.286) (compared with efavirenz) were independently associated with the development of skin rashes. Among the 1,455 patients (62.2%) with aminotransferase data both at baseline and week 4, 72 (4.9%) developed grade 2 or greater hepatotoxicity. In multiple logistic regression analysis, presence of antibody for hepatitis C virus (HCV) (AOR, 2.865; 95% CI, 1.439-5.704) or hepatitis B surface antigen (AOR, 2.397; 95% CI, 1.150-4.997), and development of skin rashes (AOR, 2.811; 95% CI, 1.051-7.521) were independently

  5. An isocratic liquid chromatography method for determining HIV non-nucleoside reverse transcriptase inhibitor and protease inhibitor concentrations in human plasma.

    Science.gov (United States)

    Weller, Dennis R; Brundage, Richard C; Balfour, Henry H; Vezina, Heather E

    2007-04-01

    An efficient, isocratic high performance liquid chromatography (HPLC) method for determining human immunodeficiency virus (HIV) non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PIs) in plasma is advantageous for laboratories participating in clinical trials and therapeutic drug monitoring (TDM) programs, or conducting small animal research. The combination of isocratic reversed phase chromatography using an S-3, 3.0 mm x 150 mm column along with low plasma volume (200 microl), rapid liquid-liquid extraction, and detection at a single wavelength (212 nm) over a short run time makes this method valuable. Within and between assay variability ranges from 0.8 to 3.5% and 1.2-6.2%, respectively. Accuracy ranges from 91.0 to 112.8% for four quality controls (50, 100, 1000, and 10,000 ng/ml) for all drugs measured (efavirenz, nevirapine, amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir).

  6. Substrate mimicry: HIV-1 reverse transcriptase recognizes 6-modified-3′-azido-2′,3′-dideoxyguanosine-5′-triphosphates as adenosine analogs

    Science.gov (United States)

    Herman, Brian D.; Schinazi, Raymond F.; Zhang, Hong-wang; Nettles, James H.; Stanton, Richard; Detorio, Mervi; Obikhod, Aleksandr; Pradère, Ugo; Coats, Steven J.; Mellors, John W.; Sluis-Cremer, Nicolas

    2012-01-01

    β-D-3′-Azido-2′,3′-dideoxyguanosine (3′-azido-ddG) is a potent inhibitor of HIV-1 replication with a superior resistance profile to zidovudine. Recently, we identified five novel 6-modified-3′-azido-ddG analogs that exhibit similar or superior anti-HIV-1 activity compared to 3′-azido-ddG in primary cells. To gain insight into their structure–activity–resistance relationships, we synthesized their triphosphate (TP) forms and assessed their ability to inhibit HIV-1 reverse transcriptase (RT). Steady-state and pre-steady-state kinetic experiments show that the 6-modified-3′-azido-ddGTP analogs act as adenosine rather than guanosine mimetics in DNA synthesis reactions. The order of potency of the TP analogs against wild-type RT was: 3′-azido-2,6-diaminopurine >3′-azido-6-chloropurine; 3′-azido-6-N-allylaminopurine > 2-amino-6-N,N-dimethylaminopurine; 2-amino-6-methoxypurine. Molecular modeling studies reveal unique hydrogen-bonding interactions between the nucleotide analogs and the template thymine base in the active site of RT. Surprisingly, the structure–activity relationship of the analogs differed in HIV-1 RT ATP-mediated excision assays of their monophosphate forms, suggesting that it may be possible to rationally design a modified base analog that is efficiently incorporated by RT but serves as a poor substrate for ATP-mediated excision reactions. Overall, these studies identify a promising strategy to design novel nucleoside analogs that exert profound antiviral activity against both WT and drug-resistant HIV-1. PMID:21914723

  7. evaluation of commercial hiv test kits used in nigeria

    African Journals Online (AJOL)

    User

    As technology evolved, screening, confirmatory and. HIV monitoring assays have greatly improved in terms of quality and speed. Coupled with this development is the unprecedented increase in the number of test kits that are available for determining. HIV status. Test kits that are simple, instrument-free simple/rapid tests ...

  8. Integrating tuberculosis/HIV treatment: an evaluation of the ...

    African Journals Online (AJOL)

    2013-01-25

    Jan 25, 2013 ... Scientific Letter: Integrating tuberculosis/HIV treatment: 478. Vol 55 No 5. SA Fam Pract 2013. Introduction. The Infectious Disease Clinic of Worcester Hospital provides antiretroviral (ARV) treatment to eligible patients who are infected with human immunodeficiency virus (HIV) in the Breede.

  9. A retrospective evaluation of proficiency testing, and rapid HIV test ...

    African Journals Online (AJOL)

    Background: Proficiency testing (PT) has been implemented as a form of External Quality Assurance (EQA) by the National HIV Reference Laboratory in Kenya since 2007 in order to monitor and improve on the quality of HIV testing and counselling HTC services. Objective: To compare concordance between National HIV ...

  10. Targeting HIV services to male migrant workers in southern Africa would not reverse generalized HIV epidemics in their home communities: a mathematical modeling analysis.

    Science.gov (United States)

    Klein, Daniel J; Eckhoff, Philip A; Bershteyn, Anna

    2015-03-01

    Migrant populations such as mine workers contributed to the spread of HIV in sub-Saharan Africa. We used a mathematical model to estimate the community-wide impact of targeting treatment and prevention to male migrants. We augmented an individual-based network model, EMOD-HIV v0.8, to include an age-dependent propensity for males to migrate. Migrants were exposed to HIV outside their home community, but continued to participate in HIV transmission in the community during periodic visits. Migrant-targeted interventions would have been transformative in the 1980s to 1990s, but post-2015 impacts were more modest. When targetable migrants comprised 2% of adult males, workplace HIV prevention averted 3.5% of community-wide infections over 20 years. Targeted treatment averted 8.5% of all-cause deaths among migrants. When migrants comprised 10% of males, workplace prevention averted 16.2% of infections in the community, one-quarter of which were among migrants. Workplace prevention and treatment acted synergistically, averting 17.1% of community infections and 11.6% of deaths among migrants. These estimates do not include prevention of secondary spread of HIV or tuberculosis at the workplace. Though cost-effective, targeting migrants cannot collapse generalized epidemics in their home communities. Such a strategy would only have been possible prior to the early 1990s. However, migrant-targeted interventions synergize with general-population expansion of HIV services. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.

  11. The Impact of Macrophage Nucleotide Pools on HIV-1 Reverse Transcription, Viral Replication, and the Development of Novel Antiviral Agents

    Directory of Open Access Journals (Sweden)

    Christina Gavegnano

    2012-01-01

    Full Text Available Macrophages are ubiquitous and represent a significant viral reservoir for HIV-1. Macrophages are nondividing, terminally differentiated cells, which have a unique cellular microenvironment relative to actively dividing T lymphocytes, all of which can impact HIV-1 infection/replication, design of inhibitors targeting viral replication in these cells, emergence of mutations within the HIV-1 genome, and disease progression. Scarce dNTPs drive rNTP incorporation into the proviral DNA in macrophages but not lymphocytes. Furthermore, the efficacy of a ribose-based inhibitor that potently inhibits HIV-1 replication in macrophages, has prompted a reconsideration of the previously accepted dogma that 2′-deoxy-based inhibitors demonstrate effective inhibition of HIV-1 replication. Additionally, higher levels of dUTP and rNTP incorporation in macrophages, and lack of repair mechanisms relative to lymphocytes, provide a further mechanistic understanding required to develop targeted inhibition of viral replication in macrophages. Together, the concentrations of dNTPs and rNTPs within macrophages comprise a distinctive cellular environment that directly impacts HIV-1 replication in macrophages and provides unique insight into novel therapeutic mechanisms that could be exploited to eliminate virus from these cells.

  12. Laboratory evaluation of the Chembio Dual Path Platform HIV-Syphilis Assay

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    Mireille B. Kalou

    2016-02-01

    Full Text Available Background: Use of rapid diagnostic tests for HIV and syphilis has increased remarkably in the last decade. As new rapid diagnostic tests become available, there is a continuous need to assess their performance and operational characteristics prior to use in clinical settings.Objectives: In this study, we evaluated the performance of the Chembio Dual Path Platform (DPP® HIV–Syphilis Assay to accurately diagnose HIV, syphilis, and HIV/syphilis co-infection.Method: In 2013, 990 serum samples from the Georgia Public Health Laboratory in Atlanta, Georgia, United States were characterised for HIV and syphilis and used to evaluate the platform. HIV reference testing combined third-generation Enzyme Immunoassay and Western Blot, whereas reference testing for syphilis was conducted by the Treponema pallidum passive particle agglutination method and the TrepSure assay. We assessed the sensitivity and specificity of the DPP assay on this panel by comparing results with the HIV and syphilis reference testing algorithms.Results: For HIV, sensitivity was 99.8% and specificity was 98.4%; for syphilis, sensitivity was 98.8% and specificity was 99.4%. Of the 348 co-infected sera, 344 (98.9% were detected accurately by the DPP assay, but 11 specimens had false-positive results (9 HIV and 2 syphilis due to weak reactivity.Conclusion: In this evaluation, the Chembio DPP HIV–Syphilis Assay had high sensitivity and specificity for detecting both HIV and treponemal antibodies. Our results indicate that this assay could have a significant impact on the simultaneous screening of HIV and syphilis using a single test device for high-risk populations or pregnant women needing timely care and treatment.

  13. Laboratory evaluation of the Chembio Dual Path Platform HIV-Syphilis Assay

    Science.gov (United States)

    Castro, Arnold; Watson, Amy; Jost, Heather; Clay, Stacy; Tun, Ye; Chen, Cheng; Karem, Kevin; Nkengasong, John N.; Ballard, Ronald; Parekh, Bharat

    2016-01-01

    Background Use of rapid diagnostic tests for HIV and syphilis has increased remarkably in the last decade. As new rapid diagnostic tests become available, there is a continuous need to assess their performance and operational characteristics prior to use in clinical settings. Objectives In this study, we evaluated the performance of the Chembio Dual Path Platform (DPP®) HIV–Syphilis Assay to accurately diagnose HIV, syphilis, and HIV/syphilis co-infection. Method In 2013, 990 serum samples from the Georgia Public Health Laboratory in Atlanta, Georgia, United States were characterised for HIV and syphilis and used to evaluate the platform. HIV reference testing combined third-generation Enzyme Immunoassay and Western Blot, whereas reference testing for syphilis was conducted by the Treponema pallidum passive particle agglutination method and the TrepSure assay. We assessed the sensitivity and specificity of the DPP assay on this panel by comparing results with the HIV and syphilis reference testing algorithms. Results For HIV, sensitivity was 99.8% and specificity was 98.4%; for syphilis, sensitivity was 98.8% and specificity was 99.4%. Of the 348 co-infected sera, 344 (98.9%) were detected accurately by the DPP assay, but 11 specimens had false-positive results (9 HIV and 2 syphilis) due to weak reactivity. Conclusion In this evaluation, the Chembio DPP HIV–Syphilis Assay had high sensitivity and specificity for detecting both HIV and treponemal antibodies. Our results indicate that this assay could have a significant impact on the simultaneous screening of HIV and syphilis using a single test device for high-risk populations or pregnant women needing timely care and treatment. PMID:28879115

  14. Isolation of a laccase with HIV-1 reverse transcriptase inhibitory activity from fresh fruiting bodies of the Lentinus edodes (Shiitake mushroom).

    Science.gov (United States)

    Sun, Jian; Wang, Hexiang; Ng, Tzi Bun

    2011-04-01

    A laccase with a molecular mass of 67 kDa and inhibitory activity toward HIV-1 reverse transcriptase (IC50 = 7.5 microM) was isolated from fresh fruiting bodies of the Lentinus edodes (Shiitake mushroom). Its characteristics were compared with those of laccases from cultured mushroom mycelia reported earlier. The laccase was unadsorbed on DEAE-cellulose, Affi-gel blue gel and CM-cellulose, but was adsorbed on Con A-Sepharose. About 50-fold purification was achieved with a 19.2% yield of the enzyme. The activity of the enzyme increased steadily from 20 degrees C to 70 degreesC. The activity disappeared after exposure to the boiling temperature for 10 min. Its optimal pH was 4 and very little enzyme activity remained at and above pH 10. The laccase inhibited HIV-1 reverse transcriptase with an IC50 of 7.5 microM, but did not demonstrate any antifungal or anti-proliferative activity.

  15. Chelation Motifs Affecting Metal-dependent Viral Enzymes: N'-acylhydrazone Ligands as Dual Target Inhibitors of HIV-1 Integrase and Reverse Transcriptase Ribonuclease H Domain.

    Science.gov (United States)

    Carcelli, Mauro; Rogolino, Dominga; Gatti, Anna; Pala, Nicolino; Corona, Angela; Caredda, Alessia; Tramontano, Enzo; Pannecouque, Christophe; Naesens, Lieve; Esposito, Francesca

    2017-01-01

    Human immunodeficiency virus type 1 (HIV-1) infection, still represent a serious global health emergency. The chronic toxicity derived from the current anti-retroviral therapy limits the prolonged use of several antiretroviral agents, continuously requiring the discovery of new antiviral agents with innovative strategies of action. In particular, the development of single molecules targeting two proteins (dual inhibitors) is one of the current main goals in drug discovery. In this contest, metal-chelating molecules have been extensively explored as potential inhibitors of viral metal-dependent enzymes, resulting in some important classes of antiviral agents. Inhibition of HIV Integrase (IN) is, in this sense, paradigmatic. HIV-1 IN and Reverse Transcriptase-associated Ribonuclease H (RNase H) active sites show structural homologies, with the presence of two Mg(II) cofactors, hence it seems possible to inhibit both enzymes by means of chelating ligands with analogous structural features. Here we present a series of N'-acylhydrazone ligands with groups able to chelate the Mg(II) hard Lewis acid ions in the active sites of both the enzymes, resulting in dual inhibitors with micromolar and even nanomolar activities. The most interesting identified N'-acylhydrazone analog, compound 18, shows dual RNase H-IN inhibition and it is also able to inhibit viral replication in cell-based antiviral assays in the low micromolar range. Computational modeling studies were also conducted to explore the binding attitudes of some model ligands within the active site of both the enzymes.

  16. Hybrid chemistry. Part 4: Discovery of etravirine-VRX-480773 hybrids as potent HIV-1 non-nucleoside reverse transcriptase inhibitors.

    Science.gov (United States)

    Wan, Zheng-Yong; Tao, Yuan; Wang, Ya-Feng; Mao, Tian-Qi; Yin, Hong; Chen, Fen-Er; Piao, Hu-Ri; De Clercq, Erik; Daelemans, Dirk; Pannecouque, Christophe

    2015-08-01

    A novel series of etravirine-VRX-480773 hybrids were designed using structure-guided molecular hybridization strategy and fusing the pharmacophore templates of etravirine and VRX-480773. The anti-HIV-1 activity and cytotoxicity was evaluated in MT-4 cell cultures. The most active hybrid compound in this series, N-(2-chlorophenyl)-2-((4-(4-cyano-2,6-dimethylphenoxy)pyrimidin-2-yl)thio)acetamide 3d (EC50=0.24 , SI>1225), was more potent than delavirdine (EC50=0.66 μM, SI>67) in the anti-HIV-1 in vitro cellular assay. Studies of structure-activity relationships established a correlation between anti-HIV activity and the substitution pattern of the acetanilide group. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. [Evaluation of an immunochromatographic fourth generation test for the rapid diagnosis of acute HIV infection].

    Science.gov (United States)

    Kawahata, Takuya; Nagashima, Mami; Sadamasu, Kenji; Kojima, Yoko; Mori, Haruyo

    2013-07-01

    The early diagnosis of human immunodeficiency virus (HIV) infection is important to provide effective antiviral treatment and to prevent transmission of HIV. One of the key issues to achieve this goal is to shorten the so-called "diagnostic window period" when the humoral immune response toward the virus is not fully developed during the acute phase of HIV-1 infection. In 2008, the Espline HIV Ag/Ab test kit (E4G, Fujirebio Inc. Japan) was marketed in Japan belonging to the fourth generation of HIV test kits characterized by its ability to detect both viral antigens (Ag) and anti-HIV-1/2 antibodies (Ab). E4G is the first and only fourth generation immunochromatographic HIV test kit approved in Japan at present. To evaluate its performance to diagnose acute HIV infection (AHI), E4G was compared with fourth generation Ag/Ab ELISA test kits, a third generation PA test kit, WB and real-time PCR for the testing of 25 AHI clinical specimens. E4G detected HIV infection in 18/25 specimens (sensitivity : 72.0%), of which the viral Ag was detected in only 2 specimens (8.0%) bearing a viral load > 10 million copies/mL. No spesimens were simultaneously reactive to both Ag and Ab against HIV. The third generation PA achieved a positive score of 17/ 25 specimens (68.0%), which was almost the same as the E4G figure. In contrast the fourth generation Ag/ Ab ELISA scored all the 25 AHI specimens positive (sensitivity : 100%). Overall, although having the merit of offering a rapid diagnostic test for HIV infection, E4G does not provide a sensitivity in AHI diagnosis superior to test kits currently available.

  18. The prevalence of resistance-associated mutations to protease and reverse transcriptase inhibitors in treatment-naïve (HIV1)-infected individuals in Casablanca, Morocco.

    Science.gov (United States)

    Bakhouch, Khadija; Oulad-Lahcen, Ahd; Bensghir, Rajae; Blaghen, Mohamed; Elfilali, Kamal Marhoum; Ezzikouri, Sayeh; Abidi, Omar; Hassar, Mohamed; Wakrim, Lahcen

    2009-06-01

    The widespread use of antiretroviral agents and the growing occurrence of HIV-1 strains resistant to these drugs have given rise to serious concerns regarding the transmission of resistant viruses to newly infected persons, which may reduce the efficacy of a first-line antiretroviral therapy. RNA was extracted from plasma samples of 98 treatment-naïve individuals with a plasma HIV RNA viral load of at least 1,000 copies/ml. Both protease (pr) and reverse transcriptase (rt) were amplified and sequenced using an automated sequencer. National Agency for AIDS Research (ANRS) and Stanford HIV database algorithms were used for interpretation of resistance data. In the protease segment, various minor mutations were present in the majority of the sequenced samples with high frequencies. Only two major mutations, M46L and V82L, were separately found in three individuals of 71 (4.2%) with one carrying both mutations. In the reverse transcriptase gene, no NNRTIs-associated resistance mutations were detected. Only one patient of 70 (1.4%) carried the F77L mutation that is associated with NRTIs resistance. Genetic subtyping revealed that 74.6% of samples were infected with subtype B, 15.5% with CRF02_AG, 4.2% with CRF01_AE, 1.4% with C, 2.8% with G and 1.4% with subtype F2. The low prevalence of major mutations associated with resistance to antiretroviral drugs (ARVs) among drug-naïve individuals studied suggests that the routine of drug resistance testing may be unnecessary for all Moroccan individuals newly diagnosed or all patients beginning antiretroviral therapy. Nevertheless, continuous surveillance is required since greater access to antiretroviral drugs is expected in Morocco.

  19. A polymorphism at position 400 in the connection subdomain of HIV-1 reverse transcriptase affects sensitivity to NNRTIs and RNaseH activity.

    Directory of Open Access Journals (Sweden)

    David W Wright

    Full Text Available Reverse transcriptase (RT plays an essential role in HIV-1 replication, and inhibition of this enzyme is a key component of HIV-treatment. However, the use of RT inhibitors can lead to the emergence of drug-resistant variants. Until recently, most clinically relevant resistance mutations were found in the polymerase domain of RT. Lately, an increasing number of resistance mutations has been identified in the connection and RNaseH domain. To further explore the role of these domains we analyzed the complete RT sequence of HIV-1 subtype B patients failing therapy. Position A/T400 in the connection subdomain is polymorphic, but the proportion of T400 increases from 41% in naïve patients to 72% in patients failing therapy. Previous studies suggested a role for threonine in conferring resistance to nucleoside RT inhibitors. Here we report that T400 also mediates resistance to non-nucleoside RT inhibitors. The susceptibility to NVP and EFV was reduced 5-fold and 2-fold, respectively, in the wild-type subtype B NL4.3 background. We show that substitution A400T reduces the RNaseH activity. The changes in enzyme activity are remarkable given the distance to both the polymerase and RNaseH active sites. Molecular dynamics simulations were performed, which provide a novel atomistic mechanism for the reduction in RNaseH activity induced by T400. Substitution A400T was found to change the conformation of the RNaseH primer grip region. Formation of an additional hydrogen bond between residue T400 and E396 may play a role in this structural change. The slower degradation of the viral RNA genome may provide more time for dissociation of the bound NNRTI from the stalled RT-template/primer complex, after which reverse transcription can resume.

  20. Evaluation of dried blood spot protocols with the Bio-Rad GS HIV Combo Ag/Ab EIA and Geenius™ HIV 1/2 Supplemental Assay.

    Science.gov (United States)

    Luo, Wei; Davis, Geoff; Li, LiXia; Shriver, M Kathleen; Mei, Joanne; Styer, Linda M; Parker, Monica M; Smith, Amanda; Paz-Bailey, Gabriela; Ethridge, Steve; Wesolowski, Laura; Owen, S Michele; Masciotra, Silvina

    2017-06-01

    FDA-approved antigen/antibody combo and HIV-1/2 differentiation supplemental tests do not have claims for dried blood spot (DBS) use. We compared two DBS-modified protocols, the Bio-Rad GS HIV Combo Ag/Ab (BRC) EIA and Geenius™ HIV-1/2 (Geenius) Supplemental Assay, to plasma protocols and evaluated them in the CDC/APHL HIV diagnostic algorithm. BRC-DBS p24 analytical sensitivity was calculated from serial dilutions of p24. DBS specimens included 11 HIV-1 seroconverters, 151 HIV-1-positive individuals, including 20 on antiretroviral therapy, 31 HIV-2-positive and one HIV-1/HIV-2-positive individuals. BRC-reactive specimens were tested with Geenius using the same DBS eluate. Matched plasma specimens were tested with BRC, an IgG/IgM immunoassay and Geenius. DBS and plasma results were compared using the McNemar's test. A DBS-algorithm applied to 348 DBS from high-risk individuals who participated in surveillance was compared to HIV status based on local testing algorithms. BRC-DBS detects p24 at a concentration 18 times higher than in plasma. In seroconverters, BRC-DBS detected more infections than the IgG/IgM immunoassay in plasma (p=0.0133), but fewer infections than BRC-plasma (p=0.0133). In addition, the BRC/Geenius-plasma algorithm identified more HIV-1 infections than the BRC/Geenius-DBS algorithm (p=0.0455). The DBS protocols correctly identified HIV status for established HIV-1 infections, including those on therapy, HIV-2 infections, and surveillance specimens. The DBS protocols exhibited promising performance and allowed rapid supplemental testing. Although the DBS algorithm missed some early infections, it showed similar results when applied to specimens from a high-risk population. Implementation of a DBS algorithm would benefit testing programs without capacity for venipuncture. Published by Elsevier B.V.

  1. Evaluation of virulence factors of Candida albicans isolated from HIV-positive individuals using HAART.

    Science.gov (United States)

    de Paula Menezes, Ralciane; de Melo Riceto, Érika Bezerra; Borges, Aércio Sebastião; de Brito Röder, Denise Von Dolingër; dos Santos Pedroso, Reginaldo

    2016-06-01

    The colonization by Candida species is one of the most important factors related to the development of oral candidiasis in HIV-infected individuals. The aim of the study was to evaluate and discuss the phospholipase, proteinase, DNAse and haemolytic activities of Candida albicans isolated from the oral cavity of HIV individuals with high efficiency antiretroviral therapy. Seventy-five isolates of C. albicans obtained from saliva samples of patients with HIV and 41 isolates from HIV-negative individuals were studied. Haemolytic activity was determined in Sabouraud dextrose agar plates containing 3% glucose and 7% sheep red cells. Culture medium containing DNA base-agar, egg yolk, and bovine albumin were used to determine DNase, phospholipase and proteinase activities, respectively. All isolates from the HIV patients group had haemolytic activity, 98% showed phospholipase activity, 92% were positive for proteinase and 32% DNAse activity. Regarding the group of indivídios HIV negative, all 41 isolates presented hemolytic activity, 90.2% showed phospholipase and proteinase activity and 12.2% were positive for DNAse. The phospholipase activity was more intense for the group of HIV positive individuals. DNase production was more frequently observed in the group of HIV-positive individuals. The percentage of isolates having DNAse activity was also significantly different between the groups of patients not using any antiretroviral therapy, those using transcriptase inhibitors and those using transcriptase inhibitor and protease inhibitor in combination. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Evaluation of rapid tests for anti-HIV detection in Brazil.

    Science.gov (United States)

    Ferreira Junior, Orlando C; Ferreira, Cristine; Riedel, Maristela; Widolin, Marcya Regina Visinoni; Barbosa-Júnior, Aristides

    2005-10-01

    This assessment in Brazil was to evaluate the performance of commercially available HIV rapid test (RT) against the gold standard testing and to establish a highly sensitive and specific RT algorithm for HIV diagnosis. A prospective, anonymous and unlinked study. An evaluation of seven commercially available RT to compare their performance against the gold standard tests for Brazil. This includes two competing enzyme immunoassays plus a Western blot for confirmation. After informed consent, whole blood samples were collected from volunteers in voluntary counselling and testing sites (n = 400), antenatal clinics (n = 500) and from HIV-positive controls in AIDS treatment centres (n = 200). Two seroconversion panels, one HIV-1 subtype (B, B', C and F) panel and an operational assay performance evaluation were also part of the study parameters. For the seven RT the clinical sensitivity ranged from 97.74 to 100% and clinical specificity from 99.43 to 100%. However, only four RT were considered acceptable after full evaluation. The two EIA had a clinical sensitivity of 100% and clinical specificity of 99.32 and 99.66%. Two RT had the same performance on the seroconversions panels as the EIA. The operational assay performance evaluation for the RT indicated that Hexagon and Capillus could not be classified as simple assays. We have provided evidence that RT assays can perform equally or better than EIA for the detection of HIV antibodies. The simplicity and rapidity of the RT warrants its utilization in an algorithm for a rapid diagnosis of HIV infection.

  3. FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation

    Science.gov (United States)

    Ruzgys, Paulius; Tamošiūnas, Mindaugas; Lukinsone, Vanesa; Šatkauskas, Saulius

    2017-09-01

    It is widely known that not all of the treated cells survive after introduction of exogenous molecules via any physical method. Therefore, it is important to develop methods that would allow simultaneous evaluation of both molecular delivery efficiency and cell viability. This study presents Förster resonance energy transfer (FRET)-based method that allows molecular transfer and cell viability evaluation in a single measurement by employing two common fluorescent dyes, namely, ethidium bromide and trypan blue. The method has been validated using cell sonoporation. The FRET-based method allows the efficiency evaluation of both reversible and irreversible sonoporation in a single experiment. Therefore, this method could be used to reduce time, labor, and material cost while improving the accuracy of evaluations.

  4. Smartphone-Imaged HIV-1 Reverse-Transcription Loop-Mediated Isothermal Amplification (RT-LAMP on a Chip from Whole Blood

    Directory of Open Access Journals (Sweden)

    Gregory L. Damhorst

    2015-09-01

    Full Text Available Viral load measurements are an essential tool for the long-term clinical care of human immunodeficiency virus (HIV-positive individuals. The gold standards in viral load instrumentation, however, are still too limited by their size, cost, and sophisticated operation for these measurements to be ubiquitous in remote settings with poor healthcare infrastructure, including parts of the world that are disproportionately affected by HIV infection. The challenge of developing a point-of-care platform capable of making viral load more accessible has been frequently approached but no solution has yet emerged that meets the practical requirements of low cost, portability, and ease-of-use. In this paper, we perform reverse-transcription loop-mediated isothermal amplification (RT-LAMP on minimally processed HIV-spiked whole blood samples with a microfluidic and silicon microchip platform, and perform fluorescence measurements with a consumer smartphone. Our integrated assay shows amplification from as few as three viruses in a ~ 60 nL RT-LAMP droplet, corresponding to a whole blood concentration of 670 viruses per μL of whole blood. The technology contains greater power in a digital RT-LAMP approach that could be scaled up for the determination of viral load from a finger prick of blood in the clinical care of HIV-positive individuals. We demonstrate that all aspects of this viral load approach, from a drop of blood to imaging the RT-LAMP reaction, are compatible with lab-on-a-chip components and mobile instrumentation.

  5. Inhibition of the ribonuclease H activity of HIV-1 reverse transcriptase by GSK5750 correlates with slow enzyme-inhibitor dissociation.

    Science.gov (United States)

    Beilhartz, Greg L; Ngure, Marianne; Johns, Brian A; DeAnda, Felix; Gerondelis, Peter; Götte, Matthias

    2014-06-06

    Compounds that efficiently inhibit the ribonuclease (RNase) H activity of the human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) have yet to be developed. Here, we demonstrate that GSK5750, a 1-hydroxy-pyridopyrimidinone analog, binds to the enzyme with an equilibrium dissociation constant (K(d)) of ~400 nM. Inhibition of HIV-1 RNase H is specific, as DNA synthesis is not affected. Moreover, GSK5750 does not inhibit the activity of Escherichia coli RNase H. Order-of-addition experiments show that GSK5750 binds to the free enzyme in an Mg(2+)-dependent fashion. However, as reported for other active site inhibitors, binding of GSK5750 to a preformed enzyme-substrate complex is severely compromised. The bound nucleic acid prevents access to the RNase H active site, which represents a possible biochemical hurdle in the development of potent RNase H inhibitors. Previous studies suggested that formation of a complex with the prototypic RNase H inhibitor β-thujaplicinol is slow, and, once formed, it dissociates rapidly. This unfavorable kinetic behavior can limit the potency of RNase H active site inhibitors. Although the association kinetics of GSK5750 remains slow, our data show that this compound forms a long lasting complex with HIV-1 RT. We conclude that slow dissociation of the inhibitor and HIV-1 RT improves RNase H active site inhibitors and may circumvent the obstacle posed by the inability of these compounds to bind to a preformed enzyme-substrate complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Free Energy-Based Virtual Screening and Optimization of RNase H Inhibitors of HIV-1 Reverse Transcriptase.

    Science.gov (United States)

    Zhang, Baofeng; D'Erasmo, Michael P; Murelli, Ryan P; Gallicchio, Emilio

    2016-09-30

    We report the results of a binding free energy-based virtual screening campaign of a library of 77 α-hydroxytropolone derivatives against the challenging RNase H active site of the reverse transcriptase (RT) enzyme of human immunodeficiency virus-1. Multiple protonation states, rotamer states, and binding modalities of each compound were individually evaluated. The work involved more than 300 individual absolute alchemical binding free energy parallel molecular dynamics calculations and over 1 million CPU hours on national computing clusters and a local campus computational grid. The thermodynamic and structural measures obtained in this work rationalize a series of characteristics of this system useful for guiding future synthetic and biochemical efforts. The free energy model identified key ligand-dependent entropic and conformational reorganization processes difficult to capture using standard docking and scoring approaches. Binding free energy-based optimization of the lead compounds emerging from the virtual screen has yielded four compounds with very favorable binding properties, which will be the subject of further experimental investigations. This work is one of the few reported applications of advanced-binding free energy models to large-scale virtual screening and optimization projects. It further demonstrates that, with suitable algorithms and automation, advanced-binding free energy models can have a useful role in early-stage drug-discovery programs.

  7. Synthesis of new 2,3-diaryl-1,3-thiazolidin-4-ones as anti-HIV agents.

    Science.gov (United States)

    Rao, Angela; Balzarini, Jan; Carbone, Anna; Chimirri, Alba; De Clercq, Erik; Monforte, Anna Maria; Monforte, Pietro; Pannecouque, Christophe; Zappalà, Maria

    2004-01-01

    Several 2,3-diaryl-1,3-thiazolidin-4-ones were synthesized and evaluated as anti-HIV agents. The results of the in vitro tests showed that some of them were highly effective inhibitors of HIV-1 replication at 30-50 nM concentrations with minimal cytotoxicity, thereby acting as non-nucleoside HIV-1 reverse transcriptase inhibitors (NNRTIs).

  8. Decrease of vitamin D concentration in patients with HIV infection on a non nucleoside reverse transcriptase inhibitor-containing regimen

    Directory of Open Access Journals (Sweden)

    Colebunders Robert

    2010-11-01

    Full Text Available Abstract Background Vitamin D is an important determinant of bone health and also plays a major role in the regulation of the immune system. Interestingly, vitamin D status before the start of highly active antiretroviral therapy (HAART has been recently associated with HIV disease progression and overall mortality in HIV-positive pregnant women. We prospectively studied vitamin D status in HIV individuals on HAART in Belgium. We selected samples from HIV-positive adults starting HAART with a pre-HAART CD4 T-cell count >100 cells/mm3 followed up for at least 12 months without a treatment change. We compared 25-hydroxyvitamin D plasma [25-(OHD] concentration in paired samples before and after 12 months of HAART. 25-(OHD levels are presented using two different cut-offs: Results Vitamin D deficiency was common before HAART, the frequency of plasma 25-(OHD concentrations below 20 ng/ml and 30 below ng/ml was 43.7% and 70.1% respectively. After 12 months on HAART, the frequency increased to 47.1% and 81.6%. HAART for 12 months was associated with a significant decrease of plasma 25-(OHD concentration (p = 0.001. Decreasing plasma 25-(OHD concentration on HAART was associated in the multivariate model with NNRTI-based regimen (p = 0.001 and lower body weight (p = 0.008. Plasma 25-(OHD concentrations decreased significantly in both nevirapine and efavirenz-containing regimens but not in PI-treated patients. Conclusions Vitamin D deficiency is frequent in HIV-positive individuals and NNRTI therapy further decreases 25-(OHD concentrations. Consequently, vitamin D status need to be checked regularly in all HIV-infected patients and vitamin D supplementation should be given when needed.

  9. Reverse total shoulder arthroplasty: a biomechanical evaluation of humeral and glenosphere hardware configuration.

    Science.gov (United States)

    Tashjian, Robert Z; Burks, Robert T; Zhang, Yue; Henninger, Heath B

    2015-03-01

    Various reverse total shoulder arthroplasty (rTSA) implant options are available for the humeral and glenosphere components. This study used a cadaveric biomechanical shoulder simulator to evaluate how hardware configurations in 2 common rTSA systems affect (1) abduction/adduction range of motion (ROM), (2) rotational ROM, and (3) forces to elevate the arm. Seven pairs of shoulders were tested on a biomechanical shoulder simulator before and after rTSA implantation. The Aequalis Reverse Shoulder (Tornier, Edina, MN, USA) and the Reverse Shoulder Prosthesis (RSP; DJO Surgical, Austin, TX, USA) were implanted in opposing shoulders. Aequalis implant options included humeral polymer insert thickness and eccentricity and glenosphere tilt. RSP implant options included glenosphere diameter and lateralization, humeral shell offset, and polymer insert depth. Both the RSP and Aequalis shifted the center of rotation inferior and medially compared with native shoulders (P Hardware configurations in rTSA have different effects on passive ROM and deltoid forces required for abduction. Identifying these changes may guide surgical decision making during rTSA placement. Copyright © 2015 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

  10. Longitudinal Evaluation of Language Impairment in Youth With Perinatally Acquired Human Immunodeficiency Virus (HIV) and Youth With Perinatal HIV Exposure

    Science.gov (United States)

    Redmond, Sean M.; Yao, Tzy-Jyun; Russell, Jonathan S.; Rice, Mabel L.; Hoffman, Howard J.; Siberry, George K.; Frederick, Toni; Purswani, Murli; Williams, Paige L.

    2016-01-01

    Background Language impairment (LI) risk is increased for perinatally acquired human immunodeficiency virus-infected (PHIV) and perinatally exposed to HIV but uninfected (PHEU) youth. This study evaluates the persistence of LI in these groups. Methods The Clinical Evaluation of Language Fundamentals was repeated on participants of the Pediatric HIV/AIDS Cohort Study Adolescent Master Protocol 18 months postbaseline. Regression models identified factors associated with change in standardized score (SC) and the resolution or development of LI. Results Of 319 participants, 112 had LI at baseline. Upon re-evaluation, SCs were highly stable and changes were similar in PHIV (n = 212) and PHEU (n = 107) participants. Those with family history of language delays had a 2.39 point lower mean increase in SCs than those without, after controlling for demographic and socioeconomic factors and baseline LI status. Among PHIV participants, CD4 count language delays was a risk factor for persistence of problems. Measures of successful HIV treatment predicted more favorable outcomes among PHIV youth. PMID:27856674

  11. Reverse Link CDMA System Capacity Evaluation for Stratospheric Platform Mobile Communications

    Directory of Open Access Journals (Sweden)

    Iskandar Iskandar

    2010-10-01

    Full Text Available We propose an analysis of reverse link CDMA multispot beam stratospheric platforms (SPF in this paper. The SPF is currently proposed as a novel wireless technology for the development of the next generation fixed and mobile communications. The geometry of this technology is different from that of the terrestrial but rather similar to the satellite based cellular system. However, evaluation on the CDMA system capacity of this technology has not been much reported. This paper addresses all possible multiple access interference analyses including the effects of channel fading and shadowing in order to evaluate the system capacity. Single SPF and multiple SPF model are evaluated under perfect power control and imperfect power control. The results indicate that in SPF systems the reverse link CDMA capacity is significantly reduced because of the power control imperfections. Moreover, in multiple SPF model the interference caused by the users in overlapped region is not trivial. We found that because of this problem the capacity is reduced for both speech and real-time data applications compared with the single SPF model even though the assumption of perfect power control can be made. In order to improve the system capacity we proposed two methods, first is to increase the minimum elevation angle definition for each platform and the second is to employ an adaptive antenna.

  12. Evaluation of the performance of the Abbott ARCHITECT HIV Ag/Ab Combo Assay.

    Science.gov (United States)

    Chavez, Pollyanna; Wesolowski, Laura; Patel, Pragna; Delaney, Kevin; Owen, S Michele

    2011-12-01

    Worldwide, many countries test for HIV infection using combination assays that simultaneously detect p24 antigen and HIV antibodies. One such assay, the ARCHITECT(®) HIV Ag/Ab Combo Assay (ARCHITECT), has recently been approved by the Food and Drug Administration (FDA) for use in the United States. To evaluate the performance of ARCHITECT on well-characterized specimens from four CDC-funded studies. We evaluated 3386 HIV-infected, 7551 HIV-uninfected, and 58 acute HIV infection (AHI) specimens. HIV-infected specimens were repeatedly reactive by enzyme immunoassay (EIA) and Western blot (WB) or positive by nucleic acid amplification testing (NAAT). HIV-uninfected specimens were EIA- and NAAT-negative. AHI specimens were seronegative or indeterminate (using antibody-based EIAs, rapid tests or WB) and NAAT-positive. All specimens were de-identified and sent to Abbott Diagnostics for testing with ARCHITECT. ARCHITECT test results were compared to original study characterizations and were used to assess overall sensitivity and specificity and also sensitivity for AHI. ARCHITECT false-positive specimens with sufficient quantity were retested. Based on results from the initial ARCHITECT test, sensitivity was 99.94% (95% confidence interval [CI]: 99.79, 99.99) and specificity was 98.78% (95% CI: 98.51-99.01). Repeat testing resulted in corrected specificity of 99.50% (95% CI: 99.31, 99.64). Also, 48 AHI specimens (83%) were detected by this screening assay. The sensitivity and specificity of the ARCHITECT combination assay are very high and most AHIs were detected by the assay. Use of Ag/Ab combination assays may improve the number of AHIs identified relative to existing FDA-approved HIV-antibody only based serologic assays, particularly in high incidence populations. Published by Elsevier B.V.

  13. Field evaluation of a dual rapid diagnostic test for HIV infection and syphilis in Lima, Peru.

    Science.gov (United States)

    Bristow, Claire C; Leon, Segundo R; Huang, Emily; Brown, Brandon J; Ramos, Lourdes B; Vargas, Silver K; Flores, Juan A; Caceres, Carlos F; Klausner, Jeffrey D

    2016-05-01

    Screening for HIV and syphilis in key populations is recommended by the WHO to reduce the morbidity, mortality and transmission associated with undiagnosed and untreated infections. Rapid point-of-care tests that can detect multiple infections with a single fingerprick whole blood specimen using a single device are gaining popularity. We evaluated the field performance of a rapid dual HIV and syphilis test in people at high risk of HIV and syphilis infections. Participants included men who have sex with men and transgender women recruited in Lima, Peru. Reference standard testing for detection of HIV and syphilis infections, conducted using blood samples from venipuncture, included Treponema pallidum particle agglutination and fourth-generation HIV enzyme immunoassay for which positive results had a confirmation HIV Western blot test. For the evaluation test, SD BIOLINE HIV/Syphilis Duo test (Standard Diagnostics, Korea), a fingerprick blood specimen was used. Sensitivity and specificity were calculated and the exact binomial method was used to determine 95% CIs. A total of 415 participants were recruited for the study. The dual test sensitivity for detection of T. pallidum infection was 89.2% (95% CI 83.5% to 93.5%) and specificity 98.8% (95% CI 96.5% to 99.8%). For detection of HIV infection, the sensitivity of the dual test was 99.1% (95% CI 94.8% to 100%) and specificity 99.4% (95% CI 97.7% to 99.9%). This high performing dual test should be considered for the use in clinical settings to increase uptake of simultaneous testing of HIV and syphilis and accelerate time to treatment for those who need it. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  14. Clinical laboratory evaluation of a reverse CAMP test for presumptive identification of Clostridium perfringens.

    OpenAIRE

    Buchanan, A G

    1982-01-01

    Ninety-six percent of Clostridium perfringens isolates from clinical specimens were reverse CAMP test positive, whereas several other Clostridium species tested were reverse CAMP test negative. C. perfringens was detected by direct inoculation of clinical specimens to reverse CAMP plates, and the reverse CAMP procedure provided reliable presumptive identification of this organism.

  15. Quantifying factors determining the rate of CTL escape and reversion during acute and chronic phases of HIV infection

    Energy Technology Data Exchange (ETDEWEB)

    Ganusov, Vitaly V [Los Alamos National Laboratory; Korber, Bette M [Los Alamos National Laboratory; Perelson, Alan S [Los Alamos National Laboratory

    2009-01-01

    Human immunodeficiency virus (HIV) often evades cytotoxic T cell (CTL) responses by generating variants that are not recognized by CTLs. However, the importance and quantitative details of CTL escape in humans are poorly understood. In part, this is because most studies looking at escape of HIV from CTL responses are cross-sectional and are limited to early or chronic phases of the infection. We use a novel technique of single genome amplification (SGA) to identify longitudinal changes in the transmitted/founder virus from the establishment of infection to the viral set point at 1 year after the infection. We find that HIV escapes from virus-specific CTL responses as early as 30-50 days since the infection, and the rates of viral escapes during acute phase of the infection are much higher than was estimated in previous studies. However, even though with time virus acquires additional escape mutations, these late mutations accumulate at a slower rate. A poor correlation between the rate of CTL escape in a particular epitope and the magnitude of the epitope-specific CTL response suggests that the lower rate of late escapes is unlikely due to a low efficacy of the HIV-specific CTL responses in the chronic phase of the infection. Instead, our results suggest that late and slow escapes are likely to arise because of high fitness cost to the viral replication associated with such CTL escapes. Targeting epitopes in which virus escapes slowly or does not escape at all by CTL responses may, therefore, be a promising direction for the development of T cell based HIV vaccines.

  16. Technology evaluation: HIV ribozyme gene therapy, Gene Shears Pty Ltd.

    Science.gov (United States)

    de Feyter, R; Li, P

    2000-06-01

    Ribozymes (catalytic RNAs) can be made to specifically cleave target RNAs that are involved in disease conditions and therefore have potential as therapeutic agents. Gene Shears Pty Ltd is developing hammerhead ribozyme technology for therapy against HIV infection, targeting either the tat gene or the RNA packaging sequence (Psi) of HIV. These ribozymes have been expressed from constructs that were introduced into hematopoietic cells in culture, thereby protecting the cells against viral infection. Two phase I clinical trials are underway to test the safety and feasibility of the approach with the anti-tat ribozyme in human subjects.

  17. Trial and error: evaluating and refining a community model of HIV testing in Australia.

    Science.gov (United States)

    Ryan, Kathleen E; Pedrana, Alisa; Leitinger, David; Wilkinson, Anna L; Locke, Peter; Hellard, Margaret E; Stoové, Mark

    2017-10-10

    The 2012 regulatory approval of HIV rapid point of care (RPOC) tests in Australia and a national strategic focus on HIV testing provided a catalyst for implementation of non-clinical HIV testing service models. PRONTO! opened in 2013 as a two-year trial delivering peer-led community-based HIV RPOC tests targeting gay, bisexual and other men who have sex with men (GBM), with the aim of increasing HIV testing frequency. Initial data suggested this aim was not achieved and, as part of a broader service evaluation, we sought to explore client acceptability and barriers to testing at PRONTO! to refine the service model. We present descriptive and thematic analyses of data from two in-depth evaluation surveys and four focus groups with PRONTO! clients focused on service acceptability, client testing history, intentions to test and barriers to testing for HIV and other sexually transmitted infections (STIs). The three novel aspects of the PRONTO! model, testing environment, rapid-testing, peer-staff, were reported to be highly acceptable among survey and focus group participants. Focus group discussions revealed that the PRONTO! model reduced anxiety associated with HIV testing and created a comfortable environment conducive to discussing sexual risk and health. However, an absence of STI testing at PRONTO!, driven by restrictions on medical subsidies for STI testing and limited funds available at the service level created a barrier to HIV testing. An overwhelming majority of PRONTO! clients reported usually testing for STIs alongside HIV and most reported plans to seek STI testing after testing for HIV at PRONTO!. When deciding where, when and what to test for, clients reported balancing convenience and relative risk and consequences for each infection as guiding their decision-making. A community-based and peer-led HIV testing model reduced previously reported barriers to HIV testing, while introducing new barriers. The absence of STI testing at PRONTO! and the need to

  18. Use of nucleoside reverse transcriptase inhibitors and risk of myocardial infarction in HIV-infected patients enrolled in the D:A:D study: a multi-cohort collaboration

    NARCIS (Netherlands)

    Sabin, Caroline A.; Worm, Signe W.; Weber, Rainer; Reiss, Peter; El-Sadr, Wafaa; Dabis, Francois; de Wit, Stephane; Law, Matthew; D'Arminio Monforte, Antonella; Friis-Møller, Nina; Kirk, Ole; Pradier, Christian; Weller, Ian; Phillips, Andrew N.; Lundgren, Jens D.; Collins, S.; Dabis, F.; d'Arminio Monforte, A.; de Wit, S.; El-Sadr, W. M.; Kirk, O.; Law, M.; Pradier, C.; Phillips, A.; Rosseau, F.; Storfer, S. P.; Weber, I.; Lundgren, J. D.; Worm, S. W.; Friis-øller, N.; Sabin, C. A.; Sjøl, A.; Sawitz, A.; Rickenbach, M.; Pezzotti, P.; Krum, E.; Gras, L.; Balestre, E.; Sundström, A.; Poll, B.; Fontas, E.; Torres, F.; Petoumenos, K.; Kjaer, J.; Hammer, S.; Neaton, J.; de Wolf, F.; Zaheri, S.; Bronsveld, W.; Hillebrand-Haverkort, M. E.; Prins, J. M.; Bos, J. C.; Eeftinck Schattenkerk, J. K. M.; Geerlings, S. E.; Godfried, M. H.; Lange, J. M. A.; van Leth, F. C.; Lowe, S. H.; van der Meer, J. T. M.; Nellen, F. J. B.; Pogány, K.; van der Poll, T.; Ruys, Th A.; Sankatsing, Raaj R.; Steingrover, R.; van Twillert, G.; van der Valk, M.; van Vonderen, M. G. A.; Vrouenraets, S. M. E.; van Vugt, M.; Wit, F. W. M. N.; van Eeden, A.; ten Veen, J. H.; van Dam, P. S.; Roos, J. C.; Brinkman, K.; Frissen, P. H. J.; Weigel, H. M.; Mulder, J. W.; van Gorp, E. C. M.; Meenhorst, P. L.; Mairuhu, A. T. A.; Veenstra, J.; Danner, S. A.; van Agtmael, M. A.; Claessen, F. A. P.; Perenboom, R. M.; Rijkeboer, A.; van Vonderen, M.; Richter, C.; van der Berg, J.; van Leusen, R.; Vriesendorp, R.; Jeurissen, F. J. F.; Kauffmann, R. H.; Koger, E. L. W.; Bravenboer, B.; ten Napel, C. H. H.; Kootstra, G. J.; Sprenger, H. G.; Miesen, W. M. A. J.; Doedens, R.; Scholvinck, E. H.; ten Kate, R. W.; van Houte, D. P. F.; Polee, M.; Kroon, F. P.; van den Broek, A. N.; van Dissel, J. T.; Schippers, E. F.; Schreij, G.; van de Geest, S.; Verbon, A.; Koopmans, P. P.; Keuter, M.; Post, F.; van der Ven, A. J. A. M.; van der Ende, M. E.; Gyssens, I. C.; van der Feltz, M.; den Hollander, J. G.; de Marie, S.; Nouwen, J. L.; Rijnders, B. J. A.; de Vries, T. E. M. S.; Juttmann, J. R.; van de Heul, C.; van Kasteren, M. E. E.; Schneider, M. M. E.; Bonten, M. J. M.; Borleffs, J. C. C.; Ellerbroek, P. M.; Hoepelman, I. M.; Jaspers, C. A. J. J.; Schouten, I.; Schurink, C. A. M.; Blok, W. L.; Tanis, A. A.; Groeneveld, P. H. P.; Salamon, R.; Beylot, J.; Dupon, M.; Le Bras, M.; Pellegrin, J. L.; Ragnaud, J. M.; Chéne, G.; Jacqmin-Gadda, H.; Thiébaut, R.; Lawson-Ayayi, S.; Lavignolle, V.; Blaizeau, M. J.; Decoin, M.; Formaggio, A. M.; Delveaux, S.; Labarerre, S.; Uwamaliya, B.; Vimard, E.; Merchadou, L.; Palmer, G.; Touchard, D.; Dutoit, D.; Pereira, F.; Boulant, B.; Morlat, P.; Bernard, N.; Bonarek, M.; Bonnet, F.; Coadou, B.; Gelie, P.; Jaubert, D.; Nouts, C.; Lacoste, D.; Dutronc, H.; Cipriano, G.; Lafarie, S.; Chossat, I.; Lacut, J. Y.; Leng, B.; Mercié, P.; Viallard, J. F.; Faure, I.; Rispal, P.; Cipriano, C.; Tchamgoué, S.; Djossou, F.; Malvy, D.; Pivetaud, J. P.; Chambon, D.; de La Taille, C.; Galperine, T.; Neau, D.; Ochoa, A.; Beylot, C.; Doutre, M. S.; Bezian, J. H.; Moreau, J. F.; Taupin, J. L.; Conri, C.; Constans, J.; Couzigou, P.; Castera, L.; Fleury, H.; Lafon, M. E.; Masquelier, B.; Pellegrin, I.; Trimoulet, P.; Moreau, F.; Mestre, C.; Series, C.; Taytard, A.; Anderson, J.; Cortossis, P.; Hoy, J.; Watson, K.; Roth, N.; Bloch, M.; Franic, T.; Baker, D.; McFarlane, R.; Carr, A.; Cooper, D.; Chuah, J.; Fankhauser, W.; Mallal, S.; Forsdyke, C.; Calvo, G.; Mateu, S.; Domingo, P.; Sambeat, M. A.; Gatel, J.; del Cacho, E.; Cadafalch, J.; Fuster, M.; Codina, C.; Sirera, G.; Vaqué, A.; Clumeck, N.; Gerard, M.; Kabeya, K.; Konopnicki, D.; Libois, A.; Payen, M. C.; van Laethem, Y.; Bartsch, G.; Thompson, G.; Wentworth, D.; Luskin-Hawk, R.; Telzak, E.; Abrams, D. I.; Cohn, D.; Markowitz, N.; Arduino, R.; Mushatt, D.; Friedland, G.; Perez, G.; Tedaldi, E.; Fisher, E.; Gordin, F.; Crane, L. R.; Sampson, J.; Baxter, J.; Mocroft, A.; Phillips, A. N.; Vetter, N.; Karpov, I.; Vassilenko, A.; Colebunders, R.; Machala, L.; Rozsypal, H.; Sedlacek, D.; Nielsen, J.; Benfield, T.; Gerstoft, J.; Katzenstein, T.; Hansen, A. B. E.; Skinhøj, P.; Pedersen, C.; Zilmer, K.; Katlama, C.; Viard, J.-P.; Girard, P.-M.; Saint-Marc, T.; Vanhems, P.; Dietrich, M.; Manegold, C.; van Lunzen, J.; Stellbrink, H.-J.; Staszewski, S.; Bieckel, M.; Goebel, F. D.; Fätkenheuer, G.; Rockstroh, J.; Schmidt, R. E.; Kosmidis, J.; Gargalianos, P.; Sambatakou, H.; Perdios, J.; Panos, G.; Filandras, A.; Banhegyi, D.; Mulcahy, F.; Yust, I.; Burke, M.; Turner, D.; Pollack, S.; Hassoun, J.; Sthoeger, Z.; Maayan, S.; Vella, S.; Chiesi, A.; Arici, C.; Pristerá, R.; Mazzotta, F.; Gabbuti, A.; Esposito, R.; Bedini, A.; Chirianni, A.; Montesarchio, E.; Vullo, V.; Santopadre, P.; Narciso, P.; Antinori, A.; Franci, P.; Zaccarelli, M.; Lazzarin, A.; Castagna, A.; Viksna, L.; Chaplinskas, S.; Hemmer, R.; Staub, T.; Bruun, J.; Maeland, A.; Ormaasen, V.; Knysz, B.; Gasiorowski, J.; Horban, A.; Prokopowicz, D.; Wiercinska-Drapalo, A.; Boron-Kaczmarsk, A.; Pynka, M.; Beniowski, M.; Mularska, E.; Trocha, H.; Antunes, A.; Mansinho, K.; Maltez, F.; Duiculescu, D.; Streinu-Cercel, A.; Vinogradova, E.; Rakhmanova, A.; Jevtovic, D.; Mokrás, M.; Staneková, D.; González-Lahoz, J.; Sanchez-Conde, M.; García-Benayas, T.; Martin-Carbonero, L.; Soriano, V.; Clotet, B.; Jou, A.; Conejero, J.; Ruiz, L.; Tural, C.; Gatell, J. M.; Miró, J. M.; Zamora, L.; Gutierrez, M.; Mateo, G.; Blaxhult, A.; Karlsson, A.; Pehrson, P.; Ledergerber, B.; Weber, R.; Francioli, P.; Telenti, A.; Hirschel, B.; Soravia-Dunand, V.; Furrer, H.; Kravchenko, E.; Chentsova, N.; Fisher, M.; Brettle, R.; Barton, S.; Johnson, A. M.; Mercey, D.; Murphy, M.; Johnson, M. A.; Weber, J.; Scullard, G.; Morfeld, L.; Thulin, G.; Akerlund, B.; Koppel, K.; Flamholc, L.; Håkangård, C.; Moroni, M.; Cargnel, A.; Merli, S.; Vigevani, G. M.; Pastecchia, C.; Morsica, G.; Caggese, L.; Moioli, C.; Mura, M. S.; Mannazzu, M.; Suter, F.; Manconi, P. E.; Piano, P.; Lo Caputo, S.; Poggio, A.; Bottari, G.; Pagano, G.; Alessandrini, A.; Scasso, A.; Abbadessa, V.; Mancuso, S.; Alberici, F.; Ruggieri, A.; Arlotti, M.; Ortolani, P.; de Lalla, F.; Tositti, G.; Cassola, G.; Piscopo, R.; Raise, E.; Ebo, F.; Soscia, F.; Tacconi, L.; Tirelli, U.; Cinelli, R.; Santoro, D.; Pusterla, L.; Carosi, G.; Torti, C.; Cadeo, G.; Bertelli, D.; Carnevale, G.; Citterio, P.; Filice, G.; Bruno, R.; Di Perri, G.; Arnaudo, I.; Caramello, P.; Orofino, G. C.; Soranzo, M. L.; Bonasso, M.; Rizzardini, G.; Melzi, S.; Chiodo, F.; Colangeli, V.; Magnani, G.; Ursitti, M.; Menichetti, F.; Martinelli, C.; Mussini, C.; Ghinelli, F.; Sighinolfi, L.; Coronado, O.; Ballardini, G.; Rizzo, E.; Montroni, M.; Braschi, M. C.; Petrelli, E.; Cioppi, A.; Cauda, R.; de Luca, A.; Petrosillo, N.; Noto, P.; Bontempo, G.; Acinapura, R.; Antonucci, G.; de Longis, P.; Lichtner, M.; Pastore, G.; Ladisa, N.; Viglietti, R.; Piazza, M.; Nappa, S.; Abrescia, N.; de Marco, M.; Colomba, A.; Prestileo, T.; de Stefano, C.; La Gala, A.; Cosco, L.; Scerbo, A.; Grima, P.; Tundo, P.; Vecchiet, J.; D'Alessandro, M.; Grisorio, B.; Ferrara, S.; Caissotti, C.; Dellamonica, P.; Bentz, L.; Bernard, E.; de Salvador-Guillouet, F.; Durant, J.; Mondain-Miton, V.; Perbost, I.; Prouvost-Keller, B.; Pugliese, P.; Rahelinirina, V.; Roger, P. M.; Vander, F.; Battegay, M.; Bernasconi, E.; Böni, J.; Buche, H.; Bürgisser, Ph; Cattacin, S.; Cavassini, M.; Dubs, R.; Egger, M.; Elzi, L.; Erb, P.; Fischer, M.; Flepp, M.; Fontana, A.; Furrer, H. J.; Gorgievski, M.; Günthard, H.; Kaiser, L.; Kind, C.; Klimkait, Th; Lauper, U.; Opravil, M.; Paccaud, F.; Pantaleo, G.; Perrin, L.; Piffaretti, J.-C.; Rudin, C.; Schmid, P.; Schüpbach, J.; Speck, R.; Trkola, A.; Vernazza, P.; Yerly, S.

    2008-01-01

    BACKGROUND: Whether nucleoside reverse transcriptase inhibitors increase the risk of myocardial infarction in HIV-infected individuals is unclear. Our aim was to explore whether exposure to such drugs was associated with an excess risk of myocardial infarction in a large, prospective observational

  19. Immunovirological response to triple nucleotide reverse-transcriptase inhibitors and ritonavir-boosted protease inhibitors in treatment-naive HIV-2-infected patients : The ACHIEV2E Collaboration Study Group

    NARCIS (Netherlands)

    Benard, Antoine; van Sighem, Ard; Taieb, Audrey; Valadas, Emilia; Ruelle, Jean; Soriano, Vicente; Calmy, Alexandra; Balotta, Claudia; Damond, Florence; Brun-Vezinet, Françoise; Chene, Geneviève; Matheron, Sophie; Schölvinck, Elisabeth H.

    2011-01-01

    BACKGROUND: Triple nucleoside reverse-transcriptase inhibitors (NRTIs) are recommended by the World Health Organization as first-line regimen in treatment-naïve HIV-2-infected patients. However, ritonavir-boosted protease inhibitor (PI/r)-containing regimens are frequently prescribed. In the absence

  20. [Screening of HIV in blood banks. Evaluation of fourth generation kits].

    Science.gov (United States)

    Canna, Fernando; Treviño, Elena; Domínguez, Claudia; Gastaldello, Rene; Barbas, Gabriela; Cudola, Analia; Irizar, Marta; Bepre, Hector; Gallego, Sandra

    2003-01-01

    Use of detection tests for p24 HIV antigen (p24Ag) in blood banks in Argentina is recommended by the Argentinean Society of Hemotherapy and Immunohematology. In the blood bank of the National University of Cordoba (Argentina), the recent implementation of the p24Ag screening test has considerably increased the cost of the battery of screening tests and its use in all blood donations has not produced the benefits expected. A 4th generation EIA was evaluated for the screening of HIV in comparison with the currently used assays in the blood bank of National University of Cordoba (3rd generation EIA + p24Ag assay). For this comparison, 11 serum samples from subjects with early HIV infection (early seroconversion period) were tested, as well as 27 serum samples from asymptomatic HIV-infected subjects and other 39 from non-HIV infected subjects. The 3rd generation EIA and the 4th generation EIA showed the same sensitivity value (100%) but the specificity of the 3rd generation EIA was higher (97.5%) comparing with 4th generation (95.1%). Besides, the p24Ag test failed to detect 2 samples from subjects with early HIV infection. These results indicate a good performance of both 3rd and 4th generation assays for screening of HIV. However, due to the lowest cost of 4th generation EIA kit, it could replace the currently used assays for HIV screening in regional blood banks. This screening assay will lead to gain in effectiveness and reduced costs until the detection of HIV RNA can be implemented in blood banks.

  1. Towards more accurate HIV testing in sub-Saharan Africa: a multi-site evaluation of HIV RDTs and risk factors for false positives.

    Science.gov (United States)

    Kosack, Cara S; Page, Anne-Laure; Beelaert, Greet; Benson, Tumwesigye; Savane, Aboubacar; Ng'ang'a, Anne; Andre, Bita; Zahinda, Jean-Paul Bn; Shanks, Leslie; Fransen, Katrien

    2017-03-24

    Although individual HIV rapid diagnostic tests (RDTs) show good performance in evaluations conducted by WHO, reports from several African countries highlight potentially significant performance issues. Despite widespread use of RDTs for HIV diagnosis in resource-constrained settings, there has been no systematic, head-to-head evaluation of their accuracy with specimens from diverse settings across sub-Saharan Africa. We conducted a standardized, centralized evaluation of eight HIV RDTs and two simple confirmatory assays at a WHO collaborating centre for evaluation of HIV diagnostics using specimens from six sites in five sub-Saharan African countries. Specimens were transported to the Institute of Tropical Medicine (ITM), Antwerp, Belgium for testing. The tests were evaluated by comparing their results to a state-of-the-art reference algorithm to estimate sensitivity, specificity and predictive values. 2785 samples collected from August 2011 to January 2015 were tested at ITM. All RDTs showed very high sensitivity, from 98.8% for First Response HIV Card Test 1-2.0 to 100% for Determine HIV 1/2, Genie Fast, SD Bioline HIV 1/2 3.0 and INSTI HIV-1/HIV-2 Antibody Test kit. Specificity ranged from 90.4% for First Response to 99.7% for HIV 1/2 STAT-PAK with wide variation based on the geographical origin of specimens. Multivariate analysis showed several factors were associated with false-positive results, including gender, provider-initiated testing and the geographical origin of specimens. For simple confirmatory assays, the total sensitivity and specificity was 100% and 98.8% for ImmunoComb II HIV 12 CombFirm (ImmunoComb) and 99.7% and 98.4% for Geenius HIV 1/2 with indeterminate rates of 8.9% and 9.4%. In this first systematic head-to-head evaluation of the most widely used RDTs, individual RDTs performed more poorly than in the WHO evaluations: only one test met the recommended thresholds for RDTs of ≥99% sensitivity and ≥98% specificity. By performing all tests

  2. Performance measurement for supply chain management and evaluation criteria determination for reverse supply chain management

    Science.gov (United States)

    Kongar, N. Elif

    2004-12-01

    Today, since customers are able to obtain similar-quality products for similar prices, the lead time has become the only preference criterion for most of the consumers. Therefore, it is crucial that the lead time, i.e., the time spent from the raw material phase till the manufactured good reaches the customer, is minimized. This issue can be investigated under the title of Supply Chain Management (SCM). An efficiently managed supply chain can lead to reduced response time for customers. To achieve this, continuous observation of supply chain efficiency, i.e., a constant performance evaluation of the current SCM is required. Widely used conventional performance measurement methods lack the ability to evaluate a SCM since the supply chain is a dynamic system that requires a more thorough and flexible performance measurement technique. Balanced Scorecard (BS) is an efficient tool for measuring the performance of dynamic systems and has a proven capability of providing the decision makers with the appropriate feedback data. In addition to SCM, a relatively new management field, namely reverse supply chain management (RSCM), also necessitates an appropriate evaluation approach. RSCM differs from SCM in many aspects, i.e., the criteria used for evaluation, the high level of uncertainty involved etc., not allowing the usage of identical evaluation techniques used for SCM. This study proposes a generic Balanced Scorecard to measure the performance of supply chain management while defining the appropriate performance measures for SCM. A scorecard prototype, ESCAPE, is presented to demonstrate the evaluation process.

  3. An evaluation of a refresher training intervention for HIV lay ...

    African Journals Online (AJOL)

    Charles Msisuka a , Ikuma Nozaki b i-nozaki@it.ncgm.go.jpinozaki@hsph.harvard.edu, Kazuhiro Kakimoto c , Motoko Seko d & Mercy M S Ulaya e

    Abstract. To address a severe shortage of human resources for health, the Zambian Ministry of Health has begun to make use of lay counsellors for HIV counselling and testing. However, their skills and knowledge rarely have been reviewed or refreshed. We conducted a two-day refresher workshop for lay counsellors to ...

  4. Evaluation of Socio-Demographic Characteristics of HIV/AIDS ...

    African Journals Online (AJOL)

    Majority (45%) were married, 38% had no formal education while, 33% were traders. The study showed a higher HIV/AIDS prevalence within the ages of 21 and 60 years and females were mostly affected. This study thus suggests a system for implementation of specific and focused educational programs among this age ...

  5. An evaluation of a refresher training intervention for HIV lay ...

    African Journals Online (AJOL)

    To address a severe shortage of human resources for health, the Zambian Ministry of Health has begun to make use of lay counsellors for HIV counselling and testing. However, their skills and knowledge rarely have been reviewed or refreshed. We conducted a two-day refresher workshop for lay counsellors to review their ...

  6. Monitoring and evaluation of sport-based HIV/ AIDS awareness ...

    African Journals Online (AJOL)

    SAHARA-J: Journal of Social Aspects of HIV/AIDS. Journal Home · ABOUT · Advanced Search · Current Issue · Archives · Journal Home > Vol 14, No 1 (2017) >. Log in or Register to get access to full text downloads.

  7. Process evaluation of school-based peer education for HIV ...

    African Journals Online (AJOL)

    In 2005, a survey was conducted among all the 27 high schools of Aden, which revealed low levels of knowledge on major prevention measures, and a high level of stigma and discrimination towards people living with HIV (PLWH). The results served as a baseline for implementing a school-based peer education ...

  8. Monitoring and evaluation of sport-based HIV/ AIDS awareness ...

    African Journals Online (AJOL)

    There are number of Non-Governmental Organisations (NGOs) in South Africa that use sport as a tool to respond to Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS), however, little is reported about the outcomes and impact of these programmes. The aim of this study is to contribute to a ...

  9. Evaluation of HIV Surveillance System in Rivers State, Nigeria ...

    African Journals Online (AJOL)

    Sony Computer (Iby)

    3Department of Neuropsychiatry/Mental Health, University of Port Harcourt Teaching Hospital,. Alakahia, PMB ... Conclusion: The HIV surveillance system in Rivers State is a useful tool for planning public health activities. ... individuals, households and nations, reducing by more than half the Gross Domestic Product (GDP).

  10. Evaluation of the Implementation of Family Life and HIV Education ...

    African Journals Online (AJOL)

    Family Life and HIV Education (FLHE) programme was introduced nationwide in Nigeria in 2003. Since then little is known about the patterns of its implementation across the states in the six geo-political zones in Nigeria. This study represents an attempt to fill this lacuna in the FLHE literature in Nigeria. Quantitative data ...

  11. Evaluation of circulating natural type 1 interferon-producing cells in HIV/GBV-C and HIV/HCV coinfected patients: a preliminary study.

    Science.gov (United States)

    Haji Molla Hoseini, Mostafa; Pourfathollah, Ali-Akbar; Mohraz, Minoo; Soheili, Zahra; Amini, Sedigheh; Aghaiepour, Mahnaz; Samiee, Shahram; Nikoogoftar, Mahin; Meshkani, Reza

    2007-11-01

    GB virus-C (GBV-C) is a flavivirus that probably influences HIV infection-associated disease among HIV/GBV-C coinfected patients and inhibits the progression of HIV infection to AIDS. To address the possibility of immune-modulating effects of GBV-C coinfection in HIV patients, we evaluated interferon-producing cells in HIV/GBV-C coinfected patients and compared them to HIV-infected patients. We performed a pilot study to enumerate interferon-producing cell count by two-color flow cytometric analysis and also for determining the frequency of ongoing GBV-C and HCV infection among patients infected with HIV. Then, 83 asymptomatic HIV-positive patients were considered for evaluation of interferon-producing cells. Eighty three patients were stratified in four groups according to the HCV and GBV-C infection status: patients infected with HCV and GBV-C (GBV-C+/HCV+), patients infected with GBV-C but not with HCV (GBV-C+/HCV-), patients infected with HCV but not with GBV-C (GBV-C-/HCV+), and patients not infected by GBV-C and HCV (GBV-C-/HCV-). GBV-C was detected in plasma samples from 15.5% of HIV-infected patients and the frequency of HCV infection was 47.5%. Interferon-producing cells in GBV-C coinfected individuals revealed a wider range of numbers; however, there was no significant difference in the interferon-producing cell count among HIV-infected individuals. It does not appear that GBV-C coinfection affects generation or redistribution of interferon-producing cells in HIV-infected patients with relatively intact immune system.

  12. K70Q adds high-level tenofovir resistance to "Q151M complex" HIV reverse transcriptase through the enhanced discrimination mechanism.

    Directory of Open Access Journals (Sweden)

    Atsuko Hachiya

    2011-01-01

    Full Text Available HIV-1 carrying the "Q151M complex" reverse transcriptase (RT mutations (A62V/V75I/F77L/F116Y/Q151M, or Q151Mc is resistant to many FDA-approved nucleoside RT inhibitors (NRTIs, but has been considered susceptible to tenofovir disoproxil fumarate (TFV-DF or TDF. We have isolated from a TFV-DF-treated HIV patient a Q151Mc-containing clinical isolate with high phenotypic resistance to TFV-DF. Analysis of the genotypic and phenotypic testing over the course of this patient's therapy lead us to hypothesize that TFV-DF resistance emerged upon appearance of the previously unreported K70Q mutation in the Q151Mc background. Virological analysis showed that HIV with only K70Q was not significantly resistant to TFV-DF. However, addition of K70Q to the Q151Mc background significantly enhanced resistance to several approved NRTIs, and also resulted in high-level (10-fold resistance to TFV-DF. Biochemical experiments established that the increased resistance to tenofovir is not the result of enhanced excision, as K70Q/Q151Mc RT exhibited diminished, rather than enhanced ATP-based primer unblocking activity. Pre-steady state kinetic analysis of the recombinant enzymes demonstrated that addition of the K70Q mutation selectively decreases the binding of tenofovir-diphosphate (TFV-DP, resulting in reduced incorporation of TFV into the nascent DNA chain. Molecular dynamics simulations suggest that changes in the hydrogen bonding pattern in the polymerase active site of K70Q/Q151Mc RT may contribute to the observed changes in binding and incorporation of TFV-DP. The novel pattern of TFV-resistance may help adjust therapeutic strategies for NRTI-experienced patients with multi-drug resistant (MDR mutations.

  13. Potent cross-reactive immune response against the wild-type and drug-resistant forms of HIV reverse transcriptase after the chimeric gene immunization.

    Science.gov (United States)

    Starodubova, Elizaveta; Boberg, Andreas; Ivanov, Alexander; Latyshev, Oleg; Petrakova, Natalia; Kuzmenko, Yulia; Litvina, Marina; Chernousov, Alexander; Kochetkov, Sergey; Karpov, Vadim; Wahren, Britta; Isaguliants, Maria G

    2010-02-23

    HIV reverse transcriptase (RT) can be considered as a target and an instrument of immunotherapy aimed at limiting the emergence and spread of drug-resistant HIV. The chimeric genes coding for the wild-type and multi-drug-resistant RT (RT1.14) fused to lysosome-associated membrane protein 1 (LAMP-1) were injected intramuscularly into BALB/c mice. The immune response was assessed by ELISpot, cytokine ELISA intracellular IFN-gamma staining, and antibody ELISA. The genes for RT- and RT1.14-LAMP fusions (RT-LAMP and RT1.14-LAMP) were immunogenic generating a mixed Th1/Th2-profile of immune response, while the wild-type RT gene induced only weak immune response. Specific secretion of Th1-cytokines increased with increasing level of RT modification: RTHIV genes may represent an immunotherapeutical supplement to antiretroviral treatment preventing the emergence of drug resistance. Copyright 2009 Elsevier Ltd. All rights reserved.

  14. Introducing Catastrophe-QSAR. Application on Modeling Molecular Mechanisms of Pyridinone Derivative-Type HIV Non-Nucleoside Reverse Transcriptase Inhibitors

    Directory of Open Access Journals (Sweden)

    Marius Lazea

    2011-12-01

    Full Text Available The classical method of quantitative structure-activity relationships (QSAR is enriched using non-linear models, as Thom’s polynomials allow either uni- or bi-variate structural parameters. In this context, catastrophe QSAR algorithms are applied to the anti-HIV-1 activity of pyridinone derivatives. This requires calculation of the so-called relative statistical power and of its minimum principle in various QSAR models. A new index, known as a statistical relative power, is constructed as an Euclidian measure for the combined ratio of the Pearson correlation to algebraic correlation, with normalized t-Student and the Fisher tests. First and second order inter-model paths are considered for mono-variate catastrophes, whereas for bi-variate catastrophes the direct minimum path is provided, allowing the QSAR models to be tested for predictive purposes. At this stage, the max-to-min hierarchies of the tested models allow the interaction mechanism to be identified using structural parameter succession and the typical catastrophes involved. Minimized differences between these catastrophe models in the common structurally influential domains that span both the trial and tested compounds identify the “optimal molecular structural domains” and the molecules with the best output with respect to the modeled activity, which in this case is human immunodeficiency virus type 1 HIV-1 inhibition. The best molecules are characterized by hydrophobic interactions with the HIV-1 p66 subunit protein, and they concur with those identified in other 3D-QSAR analyses. Moreover, the importance of aromatic ring stacking interactions for increasing the binding affinity of the inhibitor-reverse transcriptase ligand-substrate complex is highlighted.

  15. HIV-1 Reverse Transcriptase Still Remains a New Drug Target: Structure, Function, Classical Inhibitors, and New Inhibitors with Innovative Mechanisms of Actions

    Directory of Open Access Journals (Sweden)

    Francesca Esposito

    2012-01-01

    Full Text Available During the retrotranscription process, characteristic of all retroviruses, the viral ssRNA genome is converted into integration-competent dsDNA. This process is accomplished by the virus-coded reverse transcriptase (RT protein, which is a primary target in the current treatments for HIV-1 infection. In particular, in the approved therapeutic regimens two classes of drugs target RT, namely, nucleoside RT inhibitors (NRTIs and nonnucleoside RT inhibitors (NNRTIs. Both classes inhibit the RT-associated polymerase activity: the NRTIs compete with the natural dNTP substrate and act as chain terminators, while the NNRTIs bind to an allosteric pocket and inhibit polymerization noncompetitively. In addition to these two classes, other RT inhibitors (RTIs that target RT by distinct mechanisms have been identified and are currently under development. These include translocation-defective RTIs, delayed chain terminators RTIs, lethal mutagenesis RTIs, dinucleotide tetraphosphates, nucleotide-competing RTIs, pyrophosphate analogs, RT-associated RNase H function inhibitors, and dual activities inhibitors. This paper describes the HIV-1 RT function and molecular structure, illustrates the currently approved RTIs, and focuses on the mechanisms of action of the newer classes of RTIs.

  16. The mutation T477A in HIV-1 reverse transcriptase (RT restores normal proteolytic processing of RT in virus with Gag-Pol mutated in the p51-RNH cleavage site

    Directory of Open Access Journals (Sweden)

    Parniak Michael A

    2010-02-01

    Full Text Available Abstract Background The p51 subunit of the HIV-1 reverse transcriptase (RT p66/p51 heterodimer arises from proteolytic cleavage of the RT p66 subunit C-terminal ribonuclease H (RNH domain during virus maturation. Our previous work showed that mutations in the RT p51↓RNH cleavage site resulted in virus with defects in proteolytic processing of RT and significantly attenuated infectivity. In some cases, virus fitness was restored after repeated passage of mutant viruses, due to reversion of the mutated sequences to wild-type. However, in one case, the recovered virus retained the mutated p51↓RNH cleavage site but also developed an additional mutation, T477A, distal to the cleavage site. In this study we have characterized in detail the impact of the T477A mutation on intravirion processing of RT. Results While the T477A mutation arose during serial passage only with the F440V mutant background, introduction of this substitution into a variety of RT p51↓RNH cleavage site lethal mutant backgrounds was able to restore substantial infectivity and normal RT processing to these mutants. T477A had no phenotypic effect on wild-type HIV-1. We also evaluated the impact of T477A on the kinetics of intravirion Gag-Pol polyprotein processing of p51↓RNH cleavage site mutants using the protease inhibitor ritonavir. Early processing intermediates accumulated in p51↓RNH cleavage site mutant viruses, whereas introduction of T477A promoted the completion of processing and formation of the fully processed RT p66/p51 heterodimer. Conclusions This work highlights the extraordinary plasticity of HIV-1 in adapting to seemingly lethal mutations that prevent RT heterodimer formation during virion polyprotein maturation. The ability of T477A to restore RT heterodimer formation and thus intravirion stability of the enzyme may arise from increased conformation flexibility in the RT p51↓RNH cleavage site region, due to loss of a hydrogen bond associated with the

  17. HIV-1 phenotypic reverse transcriptase inhibitor drug resistance test interpretation is not dependent on the subtype of the virus backbone.

    Directory of Open Access Journals (Sweden)

    Michelle Bronze

    Full Text Available To date, the majority of HIV-1 phenotypic resistance testing has been performed with subtype B virus backbones (e.g. HXB2. However, the relevance of using this backbone to determine resistance in non-subtype B HIV-1 viruses still needs to be assessed. From 114 HIV-1 subtype C clinical samples (36 ARV-naïve, 78 ARV-exposed, pol amplicons were produced and analyzed for phenotypic resistance using both a subtype B- and C-backbone in which the pol fragment was deleted. Phenotypic resistance was assessed in resulting recombinant virus stocks (RVS for a series of antiretroviral drugs (ARV's and expressed as fold change (FC, yielding 1660 FC comparisons. These Antivirogram® derived FC values were categorized as having resistant or sensitive susceptibility based on biological cut-off values (BCOs. The concordance between resistance calls obtained for the same clinical sample but derived from two different backbones (i.e. B and C accounted for 86.1% (1429/1660 of the FC comparisons. However, when taking the assay variability into account, 95.8% (1590/1660 of the phenotypic data could be considered as being concordant with respect to their resistance call. No difference in the capacity to detect resistance associated with M184V, K103N and V106M mutations was noted between the two backbones. The following was concluded: (i A high level of concordance was shown between the two backbone phenotypic resistance profiles; (ii Assay variability is largely responsible for discordant results (i.e. for FC values close to BCO; (iii Confidence intervals should be given around the BCO's, when assessing resistance in HIV-1 subtype C; (iv No systematic resistance under- or overcalling of subtype C amplicons in the B-backbone was observed; (v Virus backbone subtype sequence variability outside the pol region does not contribute to phenotypic FC values. In conclusion the HXB2 virus backbone remains an acceptable vector for phenotyping HIV-1 subtype C pol amplicons.

  18. Process and Outcome Evaluation of an Art Therapy Program for People Living with HIV/AIDS

    Science.gov (United States)

    Feldman, Matthew B.; Betts, Donna J.; Blausey, Daniel

    2014-01-01

    Program evaluation offers an opportunity for improving the implementation and impact of art therapy. This article describes a process and outcomes evaluation of an art therapy program within the mental health services unit of a community-based organization for people living with HIV/AIDS. The aims were to assess utilization patterns and program…

  19. [Pathologic markers for evaluation of reversibility in pulmonary hypertension related to congenital heart disease].

    Science.gov (United States)

    Li, Li; Huang, Li; Chen, Guo; Huang, Shian; Liu, Chao; Wang, Hongyue; Duan, Xuejin; Wang, Qingzhi; Zhao, Ranxu; He, Jianguo

    2016-01-01

    To assess the pathologic markers for evaluation of reversibility in pulmonary hypertension (PAH) related to congenital heart disease. Twenty-eight patients with congenital heart disease complicated by PAH were subclassified into reversible pulmonary hypertension (RPAH) and irreversible pulmonary hypertension (IPAH), according to post-operative mean pulmonary artery pressure (MPAP). Pulmonary vascular lesion was analyzed according to Ruan's method. Mean medium thickness percent, mean medium area percent and pulmonary arteriolar density were measured by quantitative morphometry. Immunohistochemical study for transgelin and filamin A was carried out. Amongst the 28 cases studied, 24 were RPAH and 4 were IPAH. Of the 24 patients with RPAH, 13 (54.2%, 13/24) had pulmonary vascular lesion of grade 0, 9 (37.5%, 9/24) of grade 1 and 2 (8.3%, 2/24) of grade 2. Of the 4 patients with IPAH, 1 had lesion of grade 1, 1 of grade 2 and 2 of grade 3. Both preoperative and postoperative MPAP were higher in IPAH patients than that in RPAH patients[(53.3±23.4) mmHg versus (34.1±12.7) mmHg, P=0.020 and (35.0±8.8) mmHg versus (17.8±3.9) mmHg, Pheart disease. Mean medium thickness percent, mean medium area percent and immunoreactivity for transgelin and filamin A are useful parameters.

  20. Nonnucleoside Reverse-transcriptase Inhibitor- vs Ritonavir-boosted Protease Inhibitor-based Regimens for Initial Treatment of HIV Infection

    DEFF Research Database (Denmark)

    Borges, Álvaro H; Lundh, Andreas; Tendal, Britta

    2016-01-01

    BACKGROUND: Previous studies suggest that nonnucleoside reverse-transcriptase inhibitors (NNRTIs) cause faster virologic suppression, while ritonavir-boosted protease inhibitors (PI/r) recover more CD4 cells. However, individual trials have not been powered to compare clinical outcomes. METHODS: ...

  1. Calibrating E-values for hidden Markov models using reverse-sequence null models.

    Science.gov (United States)

    Karplus, Kevin; Karchin, Rachel; Shackelford, George; Hughey, Richard

    2005-11-15

    Hidden Markov models (HMMs) calculate the probability that a sequence was generated by a given model. Log-odds scoring provides a context for evaluating this probability, by considering it in relation to a null hypothesis. We have found that using a reverse-sequence null model effectively removes biases owing to sequence length and composition and reduces the number of false positives in a database search. Any scoring system is an arbitrary measure of the quality of database matches. Significance estimates of scores are essential, because they eliminate model- and method-dependent scaling factors, and because they quantify the importance of each match. Accurate computation of the significance of reverse-sequence null model scores presents a problem, because the scores do not fit the extreme-value (Gumbel) distribution commonly used to estimate HMM scores' significance. To get a better estimate of the significance of reverse-sequence null model scores, we derive a theoretical distribution based on the assumption of a Gumbel distribution for raw HMM scores and compare estimates based on this and other distribution families. We derive estimation methods for the parameters of the distributions based on maximum likelihood and on moment matching (least-squares fit for Student's t-distribution). We evaluate the modeled distributions of scores, based on how well they fit the tail of the observed distribution for data not used in the fitting and on the effects of the improved E-values on our HMM-based fold-recognition methods. The theoretical distribution provides some improvement in fitting the tail and in providing fewer false positives in the fold-recognition test. An ad hoc distribution based on assuming a stretched exponential tail does an even better job. The use of Student's t to model the distribution fits well in the middle of the distribution, but provides too heavy a tail. The moment-matching methods fit the tails better than maximum-likelihood methods

  2. Evaluating barriers for reverse logistics implementation under a multiple stakeholders' perspective analysis using grey decision making approach

    DEFF Research Database (Denmark)

    Bouzon, Marina; Govindan, Kannan; Rodriguez, Carlos Manuel Taboada

    2018-01-01

    In the past few decades, an interest in reverse logistics has attracted the attention of industries and also has become a subject of interest for researchers. However, while reverse logistics is becoming a mandatory element of the supply chain in developed countries particularly due to legislation...... issues, the subject is still in a state of infancy in emerging economies such as Brazil. In these connections, impediments to reverse logistics implementation must be considered and analyzed, as well as the many different perspectives from the key stakeholders for their development. The objective...... of this research is to evaluate the interrelationship among reverse logistics barriers from the perspectives of the most important stakeholders in the Brazilian context. For this purpose, a Multi-Criteria Decision Making tool named grey-based Decision Making Trial and Evaluation Laboratory (grey-DEMATEL) was used...

  3. Recycling of end-of-life reverse osmosis membranes by oxidative treatment: a technical evaluation.

    Science.gov (United States)

    Coutinho de Paula, Eduardo; Gomes, Júlia Célia Lima; Amaral, Míriam Cristina Santos

    2017-07-01

    The adverse impacts caused by the disposal of thousands of tonnes per annum of reverse osmosis (RO) membranes modules have grown dramatically around the world. The objective of this study was to evaluate the technical feasibility of recycling by chemical oxidation of end-of-life RO membranes for applications in other separation processes with specifications less rigorous. The recycling technique consisted in to cause a membrane exposition with oxidant solutions in order to remove its aromatic polyamide layer and subsequent conversion to a porous membrane. The recycling technique was evaluated by water permeability and salt rejection tests before and after the oxidative treatments. Initially, membranes' chemical cleaning and pretreatment procedures were assessed. Among factors evaluated, the oxidizing agent, its concentration and pH, associated with the oxidative treatment time, showed important influence on the oxidation of the membranes. Results showed that sodium hypochlorite and potassium permanganate are efficient agents for the membrane recycling. The great increased permeability and decreased salt rejection indicated changes on membranes' selective properties. Scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), atomic force microscopy (AFM), and contact angle characterization techniques revealed marked changes on the main membranes' physical-chemical properties, such as morphology, roughness and hydrophobicity. Reuse of produced effluents and fouling tendency of recycled membranes were also evaluated.

  4. Intimate Partner Violence and HIV-Risk Behaviors: Evaluating Avoidant Coping as a Moderator.

    Science.gov (United States)

    Weiss, Nicole H; Peasant, Courtney; Sullivan, Tami P

    2017-08-01

    Women who experience intimate partner violence (IPV) report higher rates of HIV-risk behaviors. However, few studies have examined factors that may influence the strength of the link between IPV and HIV-risk behaviors. The goal of the current study was to extend extant research by evaluating the potential moderating role of avoidant coping in this relation. Participants were 212 women currently experiencing IPV (M age = 36.63, 70.8 % African American) who were recruited from the community. Significant positive associations were found between physical, psychological, and sexual IPV severity and both avoidant coping and HIV-risk behaviors. Avoidant coping moderated the relations between both physical and psychological IPV severity and HIV-risk behaviors, such that physical and psychological IPV severity were significantly associated with HIV-risk behaviors when avoidant coping was high (but not low). Findings underscore avoidant coping as an important factor in identifying and subsequently treating IPV-victimized women vulnerable to HIV-risk behaviors.

  5. Imidazo[1,2-a]pyridin-3-amines as potential HIV-1 non-nucleoside reverse transcriptase inhibitors

    CSIR Research Space (South Africa)

    Bode, ML

    2011-06-01

    Full Text Available in this study. HN N N NH CN CN TMC-278 Figure 5. PAMPA permeabilities and AlogP values for HIV RT inhibitors. Compounds were incubated in PAMPA plates for 5 h at 37 ?C at a starting concentration of 100 ?M in the donor wells. Atenolol, pindolol... INOVA instrument. Samples were referenced against chloroform at 77.00 ppm for 13C and against tetramethylsilane at 0.00 ppm for 1H. Spin multiplicities are given as s (singlet), br s (broad singlet), d (doublet), dd (doublet of doublets), t (triplet...

  6. Phenotypic mechanism of HIV-1 resistance to 3'-azido-3'-deoxythymidine (AZT): increased polymerization processivity and enhanced sensitivity to pyrophosphate of the mutant viral reverse transcriptase.

    Science.gov (United States)

    Arion, D; Kaushik, N; McCormick, S; Borkow, G; Parniak, M A

    1998-11-10

    The multiple mutations associated with high-level AZT resistance (D67N, K70R, T215F, K219Q) arise in two separate subdomains of the viral reverse transcriptase (RT), suggesting that these mutations may contribute differently to overall resistance. We compared wild-type RT with the D67N/K70R/T215F/K219Q, D67N/K70R, and T215F/K219Q mutant enzymes. The D67N/K70R/T215F/K219Q mutant showed increased DNA polymerase processivity; this resulted from decreased template/primer dissociation from RT, and was due to the T215F/K219Q mutations. The D67N/K70R/T215F/K219Q mutant was less sensitive to AZTTP (IC50 approximately 300 nM) than wt RT (IC50 approximately 100 nM) in the presence of 0.5 mM pyrophosphate. This change in pyrophosphate-mediated sensitivity of the mutant enzyme was selective for AZTTP, since similar Km values for TTP and inhibition by ddCTP and ddGTP were noted with wt and mutant RT in the absence or in the presence of pyrophosphate. The D67N/K70R/T215F/K219Q mutant showed an increased rate of pyrophosphorolysis (the reverse reaction of DNA synthesis) of chain-terminated DNA; this enhanced pyrophosphorolysis was due to the D67N/K70R mutations. However, the processivity of pyrophosphorolysis was similar for the wild-type and mutant enzymes. We propose that HIV-1 resistance to AZT results from the selectively decreased binding of AZTTP and the increased pyrophosphorolytic cleavage of chain-terminated viral DNA by the mutant RT at physiological pyrophosphate levels, resulting in a net decrease in chain termination. The increased processivity of viral DNA synthesis may be important to enable facile HIV replication in the presence of AZT, by compensating for the increased reverse reaction rate.

  7. Evaluating social outcomes of HIV/AIDS interventions: a critical assessment of contemporary indicator frameworks

    Science.gov (United States)

    Mannell, Jenevieve; Cornish, Flora; Russell, Jill

    2014-01-01

    Introduction Contemporary HIV-related theory and policy emphasize the importance of addressing the social drivers of HIV risk and vulnerability for a long-term response. Consequently, increasing attention is being given to social and structural interventions, and to social outcomes of HIV interventions. Appropriate indicators for social outcomes are needed in order to institutionalize the commitment to addressing social outcomes. This paper critically assesses the current state of social indicators within international HIV/AIDS monitoring and evaluation frameworks. Methods We analyzed the indicator frameworks of six international organizations involved in efforts to improve and synchronize the monitoring and evaluation of the HIV/AIDS response. Our analysis classifies the 328 unique indicators according to what they measure and assesses the degree to which they offer comprehensive measurement across three dimensions: domains of the social context, levels of change and organizational capacity. Results and discussion The majority of indicators focus on individual-level (clinical and behavioural) interventions and outcomes, neglecting structural interventions, community interventions and social outcomes (e.g. stigma reduction; community capacity building; policy-maker sensitization). The main tool used to address social aspects of HIV/AIDS is the disaggregation of data by social group. This raises three main limitations. Indicator frameworks do not provide comprehensive coverage of the diverse social drivers of the epidemic, particularly neglecting criminalization, stigma, discrimination and gender norms. There is a dearth of indicators for evaluating the social impacts of HIV interventions. Indicators of organizational capacity focus on capacity to effectively deliver and manage clinical services, neglecting capacity to respond appropriately and sustainably to complex social contexts. Conclusions Current indicator frameworks cannot adequately assess the social

  8. Evaluation of membrane bioreactor for advanced treatment of industrial wastewater and reverse osmosis pretreatment

    Science.gov (United States)

    2013-01-01

    The evaluation of a membrane bioreactor (MBR) for pretreatment of reverse osmosis (RO) in order to reuse and reclamation of industrial town wastewater treatment plant was investigated in this study. Performance of MBR effluent through water quality in term of parameters such as chemical oxygen demand (COD), total suspended solids (TSS), total nitrogen (TN) and total coliform (TC) were measured. Also Silt density index (SDI) was used as indicator for RO feed water. The results of this study demonstrated that MBR produce a high quality permeate water. Approximately 75%, 98%, 74% and 99.9% removal of COD, TSS, TN and TC were recorded, respectively. Also SDI of the permeate effluent from membrane was below 3 for most of the times. It means that pilot yield a high quality treated effluent from the membrane module which can be used as RO feed water. PMID:24355199

  9. M184I/V substitutions and E138K/M184I/V double substitutions in HIV reverse transcriptase do not significantly affect the antiviral activity of EFdA.

    Science.gov (United States)

    Oliveira, Maureen; Brenner, Bluma G; Xu, Hongtao; Ibanescu, Ruxandra-Ilinca; Mesplède, Thibault; Wainberg, Mark A

    2017-11-01

    4'-Ethynyl-2-fluoro-2'-deoxyadenosine (EFdA) is a potent nucleoside analogue inhibitor of HIV that has an unusually long half-life. Cell culture selections with either EFdA or lamivudine using both subtype B and non-B clinical isolates selected the M184I/V substitutions in reverse transcriptase (RT). Unlike lamivudine, however, EFdA retained significant activity against viruses containing the M184I/V substitutions. In other clinical trials that evaluated rilpivirine together with emtricitabine in first-line therapy, the emergence of both the M184I/V and E138K mutations in RT was demonstrated. Moreover, the M184I/V and E138K substitutions were shown to be compensatory for each other with regard to both efficiency of RT activity and viral replicative capacity. This creates concern that E138K might emerge as a compensatory mutation for M184I/V in the aftermath of the use of EFdA. We wished to determine whether the E138K mutation in HIV RT together with M184I/V would compromise the activity of EFdA. Recombinant viruses containing the M184I/V and/or E138K substitutions were generated by site-directed mutagenesis and evaluated in tissue culture for susceptibility to various nucleoside compounds, including EFdA. Susceptibility to EFdA was retained in M184I/V viruses that also contained the E138K substitution. Moreover, the E138K substitution was not generated in these studies under selection pressure with EFdA. These findings help to alleviate concern that EFdA may not be active against viruses that contain both the M184I/V and E138K substitutions in RT.

  10. The evaluation of a rapid in situ HIV confirmation test in a programme with a high failure rate of the WHO HIV two-test diagnostic algorithm.

    Science.gov (United States)

    Klarkowski, Derryck B; Wazome, Joseph M; Lokuge, Kamalini M; Shanks, Leslie; Mills, Clair F; O'Brien, Daniel P

    2009-01-01

    Concerns about false-positive HIV results led to a review of testing procedures used in a Médecins Sans Frontières (MSF) HIV programme in Bukavu, eastern Democratic Republic of Congo. In addition to the WHO HIV rapid diagnostic test algorithm (RDT) (two positive RDTs alone for HIV diagnosis) used in voluntary counselling and testing (VCT) sites we evaluated in situ a practical field-based confirmation test against western blot WB. In addition, we aimed to determine the false-positive rate of the WHO two-test algorithm compared with our adapted protocol including confirmation testing, and whether weakly reactive compared with strongly reactive rapid test results were more likely to be false positives. 2864 clients presenting to MSF VCT centres in Bukavu during January to May 2006 were tested using Determine HIV-1/2 and UniGold HIV rapid tests in parallel by nurse counsellors. Plasma samples on 229 clients confirmed as double RDT positive by laboratory retesting were further tested using both WB and the Orgenics Immunocomb Combfirm HIV confirmation test (OIC-HIV). Of these, 24 samples were negative or indeterminate by WB representing a false-positive rate of the WHO two-test algorithm of 10.5% (95%CI 6.6-15.2). 17 of the 229 samples were weakly positive on rapid testing and all were negative or indeterminate by WB. The false-positive rate fell to 3.3% (95%CI 1.3-6.7) when only strong-positive rapid test results were considered. Agreement between OIC-HIV and WB was 99.1% (95%CI 96.9-99.9%) with no false OIC-HIV positives if stringent criteria for positive OIC-HIV diagnoses were used. The WHO HIV two-test diagnostic algorithm produced an unacceptably high level of false-positive diagnoses in our setting, especially if results were weakly positive. The most probable causes of the false-positive results were serological cross-reactivity or non-specific immune reactivity. Our findings show that the OIC-HIV confirmation test is practical and effective in field contexts

  11. The evaluation of a rapid in situ HIV confirmation test in a programme with a high failure rate of the WHO HIV two-test diagnostic algorithm.

    Directory of Open Access Journals (Sweden)

    Derryck B Klarkowski

    Full Text Available BACKGROUND: Concerns about false-positive HIV results led to a review of testing procedures used in a Médecins Sans Frontières (MSF HIV programme in Bukavu, eastern Democratic Republic of Congo. In addition to the WHO HIV rapid diagnostic test algorithm (RDT (two positive RDTs alone for HIV diagnosis used in voluntary counselling and testing (VCT sites we evaluated in situ a practical field-based confirmation test against western blot WB. In addition, we aimed to determine the false-positive rate of the WHO two-test algorithm compared with our adapted protocol including confirmation testing, and whether weakly reactive compared with strongly reactive rapid test results were more likely to be false positives. METHODOLOGY/PRINCIPAL FINDINGS: 2864 clients presenting to MSF VCT centres in Bukavu during January to May 2006 were tested using Determine HIV-1/2 and UniGold HIV rapid tests in parallel by nurse counsellors. Plasma samples on 229 clients confirmed as double RDT positive by laboratory retesting were further tested using both WB and the Orgenics Immunocomb Combfirm HIV confirmation test (OIC-HIV. Of these, 24 samples were negative or indeterminate by WB representing a false-positive rate of the WHO two-test algorithm of 10.5% (95%CI 6.6-15.2. 17 of the 229 samples were weakly positive on rapid testing and all were negative or indeterminate by WB. The false-positive rate fell to 3.3% (95%CI 1.3-6.7 when only strong-positive rapid test results were considered. Agreement between OIC-HIV and WB was 99.1% (95%CI 96.9-99.9% with no false OIC-HIV positives if stringent criteria for positive OIC-HIV diagnoses were used. CONCLUSIONS: The WHO HIV two-test diagnostic algorithm produced an unacceptably high level of false-positive diagnoses in our setting, especially if results were weakly positive. The most probable causes of the false-positive results were serological cross-reactivity or non-specific immune reactivity. Our findings show that the OIC-HIV

  12. Mutagenesis of the Glu-89 residue in human immunodeficiency virus type 1 (HIV-1) and HIV-2 reverse transcriptases: effects on nucleoside analog resistance.

    OpenAIRE

    Song, Q; Yang, G; Goff, S P; Prasad, V R

    1992-01-01

    A Glu-89-->Gly alteration in the human immunodeficiency virus type 1 reverse transcriptase (RT) was previously shown to result in resistance to several dideoxynucleoside analogs and to phosphonoformic acid (PFA; foscarnet). This residue was altered to Ala, Val, Ser, Thr, Gln, Asp, Asn, or Lys, and the ddGTP and PFA sensitivities of the mutant RTs were measured. Replacements with Ala, Gly, Val, and Thr led to resistance to inhibition by ddGTP, while mutants with amino acid Ser, Gln, Asn, Asp, ...

  13. Evaluating Safer Conception Options for HIV-Serodiscordant Couples (HIV-Infected Female/HIV-Uninfected Male: A Closer Look at Vaginal Insemination

    Directory of Open Access Journals (Sweden)

    Okeoma Mmeje

    2012-01-01

    Full Text Available HIV serodiscordant couples represent at least half of all HIV-affected couples worldwide. Many of these couples have childbearing desires. Safer methods of conception may allow for pregnancy while minimizing the risk of sexual transmission of HIV. In serodiscordant partnerships with an HIV-infected female and HIV-uninfected male, vaginal insemination of a partner's semen during the fertile period coupled with 100% condom use may be the safest method of conception.

  14. Is the HIV sentinel surveillance system adequate in China? Findings from an evaluation of the national HIV sentinel surveillance system

    Directory of Open Access Journals (Sweden)

    Marc Bulterys

    2012-11-01

    Full Text Available Background: An external evaluation was conducted to assess the performance of the national HIV sentinel surveillance system (HSS, identify operational challenges at national and local levels and provide recommendations for improvement.Methods: The United States Centers for Disease Control and Prevention’s (CDC Updated Guidelines for Evaluating Public Health Surveillance Systems were followed to assess the key attributes of HSS. Comprehensive assessment activities were conducted, including: using a detailed checklist to review surveillance guidelines, protocols and relevant documents; conducting self-administered, anonymous surveys with 286 local China CDC staff; and carrying out field observations in 32 sentinel sites in four provinces.Results: China has built an extensive HSS with 1888 sentinel sites to monitor HIV epidemic trends by population groups over time. The strengths of HSS lie in its flexibility, simplicity, usefulness and increase in coverage in locations and populations. With its rapid expansion in 2010, HSS faces challenges in maintaining acceptability, timeliness, data quality, representativeness and sustainability.Recommendations: Implementation of the national guidelines should be standardized by strengthening training, monitoring and supervision of all staff involved, including community-based organizations. National surveillance guidelines need to be revised to strengthen data quality and representativeness, particularly to include specific instructions on HIV testing result provision, collection of identifying information, sample size and sampling methods particularly for men who have sex with men (MSM, collection of refusal information, and data interpretation. Sustainability of China’s HSS could be strengthened by applying locally tailored surveillance strategies, strengthening coordination and cooperation among government agencies and ensuring financial and human resources.

  15. Evaluating HIV prevention strategies for populations in key affected groups: The example of Cabo Verde

    Science.gov (United States)

    Monteiro, João Filipe G.; Galea, Sandro; Flanigan, Timothy; Monteiro, Maria de Lourdes; Friedman, Samuel R.; Marshall, Brandon DL

    2015-01-01

    Objectives We used an individual-based model to evaluate the effects of hypothetical prevention interventions on HIV incidence trajectories in a concentrated, mixed epidemic setting from 2011 to 2021, and using Cabo Verde as an example. Methods Simulations were conducted to evaluate the extent to which early HIV treatment and optimization of care, HIV testing, condom distribution, and substance abuse treatment could eliminate new infections (i.e., reduce incidence to less than 10 cases per 10,000 person-years) among non-drug users, female sex workers (FSW), and people who use drugs (PWUD). Results Scaling up all four interventions resulted in the largest decreases in HIV, with estimates ranging from 1.4 (95%CI:1.36–1.44) per 10,000 person-years among non-drug users to 8.2 (95%CI:7.8–8.6) per 10,000 person-years among PWUD in 2021. Intervention scenarios targeting FWS and PWUD also resulted in HIV incidence estimates at or below 10 per 10,000 person-years by 2021 for all population sub-groups. Conclusions Our results suggest that scaling up multiple interventions among entire population is necessary to achieve elimination. However, prioritizing key populations with this combination prevention strategy may also result in a substantial decrease in total incidence. PMID:25838121

  16. Evaluation of Xpert HIV-1 Qual assay for resolution of HIV-1 infection in samples with negative or indeterminate Geenius HIV-1/2 results.

    Science.gov (United States)

    Michaeli, Michal; Wax, Marina; Gozlan, Yael; Rakovsky, Aviya; Mendelson, Ella; Mor, Orna

    2016-03-01

    Diagnosis of HIV infection is a multistage algorithm. Following screening with 4(th) generation combination immunoassay, confirmation of HIV infection is performed with an antibody assay that differentiates HIV-1 from HIV-2 infection. In the newly updated algorithm, samples that are nonreactive or indeterminate in the differentiation assay are to be tested with an HIV-1 nucleic acid amplification (NAAT) test for resolution. Xpert HIV-1 Qual is a new NAAT assay approved for the identification of HIV infection in whole and dried blood. To assess the performance of Xpert HIV-1 Qual supplementary assay in resolving the clinical status of serum samples reactive by 4(th) generation immunoassays and indeterminate or negative by Geenius HIV-1/2 confirmatory assay. In a retrospective study, samples from 97 individuals for whom the true HIV-1 status was already known (by follow-up samples) and which were negative or indeterminate by HIV-1/2 Geenius assay were tested with Xpert Qual HIV-1 assay. Xpert Qual assay correctly classified all 97 samples from HIV-1 positive (n=49) and negative (n=48) individuals. The sensitivity and specificity of Xpert Qual when using the true HIV status as a reference were 100% (92.7-100% at 95% confidence interval [CI] and 92.6-100% at 95% CI, respectively). Applying Xpert Qual HIV-1 assay in the new HIV multi-stage diagnostic algorithm correctly classified 100% of HIV-1 infections including 49 from HIV-1 carriers who have not yet seroconverted. With this assay the total time required for acute HIV diagnosis could be significantly reduced. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Evaluation of lorcaserin on progression of prediabetes to type 2 diabetes and reversion to euglycemia.

    Science.gov (United States)

    Nesto, Richard; Fain, Randi; Li, Yuhan; Shanahan, William

    2016-05-01

    Lorcaserin is a selective 5-HT2C (5-hydroxytryptamine 2C) receptor agonist indicated for weight management. Here, we assess the impact of lorcaserin on progression from prediabetes to type 2 diabetes (T2D) and on reversion from prediabetes to euglycemia. This is a post hoc analysis of pooled data from two Phase 3 studies, BLOOM and BLOSSOM (N = 6136), evaluating the impact of lorcaserin on weight and glycemic parameters over 52 weeks in the subpopulation of obese/overweight subjects with prediabetes, alternately defined by fasting plasma glucose (FPG) 100-125 mg/dl or glycated hemoglobin (HbA1c) 5.7-6.4% at baseline. At Week 52, in the subpopulation with prediabetes, nearly twice as many lorcaserin-treated subjects achieved ≥5% weight loss versus placebo (HbA1c: 55.6% vs. 27.5%, p lorcaserin-treated subjects progressed to T2D versus placebo based on HbA1c (lorcaserin 3.2%, placebo 5.0%, p = 0.032) but not FPG (lorcaserin 1.6%, placebo 2.6%, p = 0.227). A significantly greater proportion of lorcaserin-treated subjects versus placebo also reverted to euglycemia based on both HbA1c (lorcaserin 40%, placebo 29.5%, p lorcaserin 52.4%, placebo 46.5%, p = 0.047). In subjects with prediabetes, lorcaserin may contribute to weight loss and improve glycemic parameters, and thus may help with preventing progression to T2D and promoting reversion to euglycemia. www.clinicaltrials.gov identifiers are NCT00395135 (BLOOM) and NCT00603902 (BLOSSOM).

  18. A reverse genetics cell-based evaluation of genes linked to healthy human tissue age.

    Science.gov (United States)

    Crossland, Hannah; Atherton, Philip J; Strömberg, Anna; Gustafsson, Thomas; Timmons, James A

    2017-01-01

    We recently developed a binary (i.e., young vs. old) classifier using human muscle RNA profiles that accurately distinguished the age of multiple tissue types. Pathway analysis did not reveal regulators of these 150 genes, so we used reverse genetics and pharmacologic methods to explore regulation of gene expression. Using small interfering RNA, well-studied age-related factors (i.e., rapamycin, resveratrol, TNF-α, and staurosporine), quantitative real-time PCR and clustering analysis, we studied gene-gene interactions in human skeletal muscle and renal epithelial cells. Individual knockdown of 10 different age genes yielded a consistent pattern of gene expression in muscle and renal cells, similar to in vivo. Potential epigenetic interactions included HIST1H3E knockdown, leading to decreased PHF19 and PCDH9, and increased ICAM5 in muscle and renal cells, while ICAM5 knockdown reduced HIST1H3E expression. Resveratrol, staurosporine, and TNF-α significantly regulated the in vivo aging genes, while only rapamycin perturbed the healthy-age gene expression signature in a manner consistent with in vivo. In vitro coordination of gene expression for this in vivo tissue age signature indicates a degree of direct coordination, and the observed link with mTOR activity suggests a direct link between a robust biomarker of healthy neuromuscular age and a major axis of life span in model systems.-Crossland, H., Atherton, P. J., Strömberg, A., Gustafsson, T., Timmons, J. A. A reverse genetics cell-based evaluation of genes linked to healthy human tissue age. © The Author(s).

  19. Mutagenesis of the Glu-89 residue in human immunodeficiency virus type 1 (HIV-1) and HIV-2 reverse transcriptases: effects on nucleoside analog resistance.

    Science.gov (United States)

    Song, Q; Yang, G; Goff, S P; Prasad, V R

    1992-12-01

    A Glu-89-->Gly alteration in the human immunodeficiency virus type 1 reverse transcriptase (RT) was previously shown to result in resistance to several dideoxynucleoside analogs and to phosphonoformic acid (PFA; foscarnet). This residue was altered to Ala, Val, Ser, Thr, Gln, Asp, Asn, or Lys, and the ddGTP and PFA sensitivities of the mutant RTs were measured. Replacements with Ala, Gly, Val, and Thr led to resistance to inhibition by ddGTP, while mutants with amino acid Ser, Gln, Asn, Asp, or Lys displayed only moderate or no resistance. A similar result was obtained with inhibition by PFA, except that the Asp-89 mutant also displayed resistance. Furthermore, the introduction of Glu-89-->Gly alteration into the RT of human immunodeficiency virus type 2 likewise rendered it resistant to both ddGTP and PFA.

  20. Prevention of mother to child transmission of HIV: evaluation of a pilot programme in a district hospital in rural Zimbabwe.

    Science.gov (United States)

    Perez, Freddy; Orne-Gliemann, Joanna; Mukotekwa, Tarisai; Miller, Anna; Glenshaw, Monica; Mahomva, Agnes; Dabis, François

    2004-11-13

    Zimbabwe has one of the highest rates of HIV seroprevalence in the world. In 2001 only 4% of women and children in need of services for prevention of mother to child transmission of HIV were receiving them. Pilot implementation of the first programme for prevention of mother to child transmission of HIV in rural Zimbabwe. 120 bed district hospital in Buhera district (285,000 inhabitants), Manicaland, Zimbabwe. Programme uptake indicators monitored for 18 months; impact of policy evaluated by assessing up-scaling of programme. Voluntary counselling and testing services for HIV were provided in the hospital antenatal clinic. Women identified as HIV positive and informed of their serostatus and their newborn were offered a single dose antiretroviral treatment of nevirapine; mother-child pairs were followed up through routine health services. Nursing staff and social workers were trained, and community mobilisation was conducted. No services for prevention of mother to child transmission of HIV were available at baseline. Within 18 months, 2298 pregnant women had received pretest counselling, and the acceptance of HIV testing reached 93.0%. Of all 2137 women who had an HIV test, 1588 (74.3%) returned to collect their result; 326 of the 437 HIV positive women diagnosed had post-test counselling, and 104 (24%) mother-child pairs received nevirapine prophylaxis. Minimum staffing, an enhanced training programme, and involvement of district health authorities are needed for the implementation and successful integration of services for prevention of mother to child transmission of HIV. Voluntary counselling and testing services are important entry points for HIV prevention and care and for referral to community networks and medical HIV care services. A district approach is critical to extend programmes for prevention of mother to child transmission of HIV in rural settings. The lessons learnt from this pilot programme have contributed to the design of the national expansion

  1. Process Evaluation of HIV Prevention Peer Groups in Malawi: A Look inside the Black Box

    Science.gov (United States)

    McCreary, Linda L.; Kaponda, Chrissie P. N.; Kafulafula, Ursula K.; Ngalande, Rebecca C.; Kumbani, Lily C.; Jere, Diana L. N.; Norr, James L.; Norr, Kathleen F.

    2010-01-01

    This paper reports the process evaluation of a peer group intervention for human immunodeficiency virus (HIV) prevention which had positive outcomes for three target groups in Malawi: rural adults, adolescents and urban hospital workers. The six-session intervention was delivered to small groups of 10-12 participants by 85 trained volunteer peer…

  2. Evaluation of the performance of HIV1 & 2 one-step selftest kit for ...

    African Journals Online (AJOL)

    further confirmed using New Lav Blot 1 western blot kit (BIO-RAD; 3, Boulevard Raymond Poincare 92430 MARNES LA COQUETTE- FRANCE). These samples were screened using the HIV1 & 2 one-step self-test kit (Bremancos Diagnostics Inc. BDI with lot Number 0141503) to evaluate its performance. Whole blood ...

  3. A six year evaluation of cracked teeth diagnosed with reversible pulpitis: treatment and prognosis.

    Science.gov (United States)

    Krell, Keith V; Rivera, Eric M

    2007-12-01

    The purpose of this investigation was to report on the clinical outcomes of cracked teeth diagnosed with reversible pulpitis (RP). Eight thousand one hundred seventy-five patients referred for evaluation and treatment during a 6-year period had medical and dental histories, radiographs, pulpal and periapical diagnosis, periodontal probings, direct identification of crack(s) with transillumination, and biting responses on various cusps recorded. All data were stored daily in a database. All cases were treatment planned according to the pulpal and periapical diagnosis. Cases with RP were treatment planned for crowns only, regardless of periapical diagnosis. All patients were recalled at 1 year unless root canal treatment was needed before the anniversary. Results indicated that cracks were identified in 9.7% (796 of 8175) of all teeth evaluated during this time period. Of 127 patients specifically diagnosed with RP, 27 converted to irreversible pulpitis (N = 21) in 58 days or to necrotic pulp (N = 6) in 149 days. To date, none of the original remaining 100 cases of RP have required root canal treatment. The outcomes of this study suggest that if a marginal ridge crack is identified early enough in teeth with a diagnosis of RP and a crown is placed, root canal treatment will be necessary in about 20% of these cases within a 6-month period.

  4. Evaluation of reverse electrodialysis system with various compositions of natural resources

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Kil Sung; Park, Byung Ho; Kim, Duk Han; Kim, Dae Joong [Dept. of Mechanical Engineering, Sogang University, Seoul (Korea, Republic of)

    2015-06-15

    Salinity gradient power (SGP) has attracted significant attention because of its high potential. In this study, we evaluate reverse electrodialysis (RED) with various compositions of available resources. The polarization curve (I-V characteristics) shows linear behavior, and therefore the power density curve has a parabolic shape. We measure the power density with varying compartment thicknesses and inlet flow rates. The gross power density increases with decreasing compartment thickness and increasing flow rate. The net power density, which is the gross power density minus the pumping power, has a maximum value at a compartment thickness of 0.2 mm and an inlet flow rate of 22.5 mL/min. The power density in RED is also evaluated with compositions of desalination brines, seawater, river water, wastewater, and brackish water. A maximum power density of 1.75W/m{sup 2} is obtained when brine discharged from forward osmosis (FO) and river water are used as the concentrated and the diluted solutions, respectively.

  5. Evaluation of Various Real-Time Reverse Transcription Quantitative PCR Assays for Norovirus Detection.

    Science.gov (United States)

    Yoo, Ju Eun; Lee, Cheonghoon; Park, SungJun; Ko, GwangPyo

    2017-04-28

    Human noroviruses are widespread and contagious viruses causing nonbacterial gastroenteritis. Real-time reverse transcription quantitative PCR (real-time RT-qPCR) is currently the gold standard for the sensitive and accurate detection of these pathogens and serves as a critical tool in outbreak prevention and control. Different surveillance teams, however, may use different assays, and variability in specimen conditions may lead to disagreement in results. Furthermore, the norovirus genome is highly variable and continuously evolving. These issues necessitate the re-examination of the real-time RT-qPCR's robustness in the context of accurate detection as well as the investigation of practical strategies to enhance assay performance. Four widely referenced real-time RT-qPCR assays (Assays A-D) were simultaneously performed to evaluate characteristics such as PCR efficiency, detection limit, and sensitivity and specificity with RT-PCR, and to assess the most accurate method for detecting norovirus genogroups I and II. Overall, Assay D was evaluated to be the most precise and accurate assay in this study. A ZEN internal quencher, which decreases nonspecific fluorescence during the PCR, was added to Assay D's probe, which further improved the assay performance. This study compared several detection assays for noroviruses, and an improvement strategy based on such comparisons provided useful characterizations of a highly optimized real-time RT-qPCR assay for norovirus detection.

  6. A template-dependent dislocation mechanism potentiates K65R reverse transcriptase mutation development in subtype C variants of HIV-1.

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    Dimitrios Coutsinos

    Full Text Available Numerous studies have suggested that the K65R reverse transcriptase (RT mutation develops more readily in subtype C than subtype B HIV-1. We recently showed that this discrepancy lies partly in the subtype C template coding sequence that predisposes RT to pause at the site of K65R mutagenesis. However, the mechanism underlying this observation and the elevated rates of K65R development remained unknown. Here, we report that DNA synthesis performed with subtype C templates consistently produced more K65R-containing transcripts than subtype B templates, regardless of the subtype-origin of the RT enzymes employed. These findings confirm that the mechanism involved is template-specific and RT-independent. In addition, a pattern of DNA synthesis characteristic of site-specific primer/template slippage and dislocation was only observed with the subtype C sequence. Analysis of RNA secondary structure suggested that the latter was unlikely to impact on K65R development between subtypes and that Streisinger strand slippage during DNA synthesis at the homopolymeric nucleotide stretch of the subtype C K65 region might occur, resulting in misalignment of the primer and template. Consequently, slippage would lead to a deletion of the middle adenine of codon K65 and the production of a -1 frameshift mutation, which upon dislocation and realignment of the primer and template, would lead to development of the K65R mutation. These findings provide additional mechanistic evidence for the facilitated development of the K65R mutation in subtype C HIV-1.

  7. Quantification of 8 HIV-protease inhibitors and 2 nonnucleoside reverse transcriptase inhibitors by ultra-performance liquid chromatography with diode array detection.

    Science.gov (United States)

    Elens, Laure; Veriter, Sophie; Di Fazio, Vincent; Vanbinst, Roger; Boesmans, Daniel; Wallemacq, Pierre; Haufroid, Vincent

    2009-01-01

    Most HPLC-UV methods for therapeutic drug monitoring of anti-HIV drugs have long run times, which reduce their applicability for high-throughput analysis. We developed an ultra-performance liquid chromatography (UPLC)-diode array detection method for the simultaneous quantification of the HIV-protease inhibitors (PIs) amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, saquinavir, and tipranavir (TPV), and the nonnucleoside reverse transcriptase inhibitors (NNRTIs) efavirenz and nevirapine. Solid-phase extraction of 1 mL plasma was performed with Waters HLB cartridges. After 3 wash steps, we eluted the drugs with methanol, evaporated the alcohol, and reconstituted the residue with 50 microL methanol. We injected a 4-microL volume into the UPLC system (Waters ACQUITY UPLC BEH C8 column maintained at 60 degrees C) and used a linear gradient of 50 mmol/L ammonium acetate and 50 mmol/L formic acid in water versus acetonitrile to achieve chromatographic separation of the drugs and internal standard (A-86093). Three wavelengths (215, 240, and 260 nm) were monitored. All drugs were eluted within 15 min. Calibration curves with concentrations of 0.025-10 mg/L (1.875-75 mg/L for TPV) showed coefficients of determination (r(2)) between 0.993 and 0.999. The lower limits of quantification were well below the trough concentrations reported in the literature. Inter- and intraassay CVs and the deviations between the nominal and measured concentrations were <15%. The method was validated by successful participation in an international interlaboratory QC program. This method allows fast and simultaneous quantification of all commercially available PIs and NNRTIs for therapeutic drug monitoring.

  8. A Full-Coordinate Model of the Polymerase Domain of HIV-I Reverse Transcriptase and its Interaction With a Nucleic Acid Substrate.

    Science.gov (United States)

    Setlik, Robert F; Meyer, David J; Shibata, Masayuki; Roskwitalski, Raymond; Ornstein, Rick L; Rein, Robert

    1994-08-01

    Abstract We present a full-coordinate model of residues 1-319 of the polymerase domain of HIV-I reverse transcriptase. This model was constructed from the x-ray crystallographic structure of Jacobo-Molina et al. (Jacobo-Molina et al., P.N.A.S. USA 90, 6320-6324 (1993)) which is currently available to the degree of C- coordinates. The backbone and side-chain atoms were constructed using the MAXSPROUT suite of programs (L. Holm and C. Sander, Mol. Biol. 218, 183-194 (1991)) and refined through molecular modeling. A seven base pair A-form dsDNA was positioned in the nucleic acid binding cleft to represent the template-primer complex. The orientation of the template-primer complex in the nucleic acid binding cleft was guided by the positions of phosphorus atoms in the crystal structure. Two magnesium ions were placed in the active site in order to better understand the polymerization mechanism. The positions of metal ions in a number of structures guided the placement of ions in this study (L.S. Beese and T.A. Steitz, EMBO. J10, 25-33 (1991); T.A. Steitz and J.A. Steitz, P.N.A.S. USA 90, 6498-6502 (1993); D.L. Sloan et al., J. Biol. Chem. 250, 8913-8920 (1975); R.F. Setlik et al., J. Biomol. Str. Dyn. 10, 945-972 (1993)). The geometry of the active site allowed metal ions to be bound to Asp 110 and Asp 186 of the catalytic triad. However, due to spacial constraints, Asp 185 was found unable to bind to a metal ion. Due to its proximity to the attacking 3'OH group of the 3' terminal residue of the primer strand, it is proposed that this residue acts as a general base which abstracts a proton from the attacking group. Based on the locations of these metal ions with respect to the attacking group of the 3' end of the primer strand and to an incoming dTTP placed in the active site, we propose roles for the magnesium ions and discuss a mechanism through which chain elongation occurs. Also reported are the interactions between the polymerase domain and the template

  9. HIV treatment as prevention: principles of good HIV epidemiology modelling for public health decision-making in all modes of prevention and evaluation.

    Directory of Open Access Journals (Sweden)

    Wim Delva

    Full Text Available Public health responses to HIV epidemics have long relied on epidemiological modelling analyses to help prospectively project and retrospectively estimate the impact, cost-effectiveness, affordability, and investment returns of interventions, and to help plan the design of evaluations. But translating model output into policy decisions and implementation on the ground is challenged by the differences in background and expectations of modellers and decision-makers. As part of the PLoS Medicine Collection "Investigating the Impact of Treatment on New HIV Infections"--which focuses on the contribution of modelling to current issues in HIV prevention--we present here principles of "best practice" for the construction, reporting, and interpretation of HIV epidemiological models for public health decision-making on all aspects of HIV. Aimed at both those who conduct modelling research and those who use modelling results, we hope that the principles described here will become a shared resource that facilitates constructive discussions about the policy implications that emerge from HIV epidemiology modelling results, and that promotes joint understanding between modellers and decision-makers about when modelling is useful as a tool in quantifying HIV epidemiological outcomes and improving prevention programming.

  10. A Mathematical Model of Antiretroviral Therapy Evaluation for HIV Type 1

    Science.gov (United States)

    Raimundo, Silvia Martorano; Venturino, Ezio; Mo Yang, Hyun

    2009-09-01

    Treating HIV-infected patients with a combination of several antiretroviral drugs can lead to emergence of the drug-resistant strain. This work proposes a mathematical model to evaluate the emergence of HIV-1 drug resistant during antiretroviral therapy. The model assumes that all susceptible individuals who can be infected by the wildtype strain (sensible to the treatment) or by drug-resistant virus receive antiretroviral therapy. Patients on treatment regimen can evolve to a state of success or failure and for the individuals in therapeutic fail the therapeutic schema is changed. The analysis of system is performed. The existence and stability of the steady states are considered. We address an analytical expression for the reproductive number in a community where antiretroviral therapy are widely used to treat HIV and where both drug sensitive and drug resistant strains are co-circulating.

  11. Visceral Leishmaniasis/HIV co-infection in northeast Brazil: evaluation of outcome.

    Science.gov (United States)

    Távora, Lara Gurgel Fernandes; Nogueira, Marina Bizerril; Gomes, Sofia Teixeira

    2015-01-01

    Since the beginning of the HIV burden, Visceral Leishmaniasis (VL)/HIV co-infection has been diagnosed not only in areas where VL is endemic (Latin America, India, Asia, Southern Europe), but also in North America, were it is considered an opportunistic disease. Clinical presentation, diagnostic tests sensitivity and treatment response in this population differs from VL alone. To evaluate factors related to an unfavorable outcome in patients with VL/HIV diagnosis in a reference center in northeast Brazil. Co-infected patients, diagnosed from 2010 to 2012, were included. Data from medical records were collected until one year after VL treatment completion. Forty-two HIV-infected patients were included in the study. Anemia, leukopenia, and thrombocytopenia were present in 95%, 70.7%, and 63.4%, respectively. Mean T CD4+ (LTCD4) lymphocyte count was 183 cells/dL. Highly active antiretroviral therapy (HAART) was being used by 54.7% of cases. A favorable outcome was seen in 71.4% of cases. Recurrence of VL occurred in nine patients and deaths were secondary to infectious complications (3/42 patients). Very low LTCD4 count (LTCD4 count at presentation was associated with unfavorable outcome in VL/HIV patients. Copyright © 2015 Elsevier Editora Ltda. All rights reserved.

  12. Visceral Leishmaniasis/HIV co-infection in northeast Brazil: evaluation of outcome

    Directory of Open Access Journals (Sweden)

    Lara Gurgel Fernandes Távora

    Full Text Available ABSTRACT Since the beginning of the HIV burden, Visceral Leishmaniasis (VL/HIV co-infection has been diagnosed not only in areas where VL is endemic (Latin America, India, Asia, Southern Europe, but also in North America, were it is considered an opportunistic disease. Clinical presentation, diagnostic tests sensitivity and treatment response in this population differs from VL alone. OBJECTIVES: To evaluate factors related to an unfavorable outcome in patients with VL/HIV diagnosis in a reference center in northeast Brazil. METHODS: Co-infected patients, diagnosed from 2010 to 2012, were included. Data from medical records were collected until one year after VL treatment completion. RESULTS: Forty-two HIV-infected patients were included in the study. Anemia, leukopenia, and thrombocytopenia were present in 95%, 70.7%, and 63.4%, respectively. Mean T CD4+ (LTCD4 lymphocyte count was 183 cells/dL. Highly active antiretroviral therapy (HAART was being used by 54.7% of cases. A favorable outcome was seen in 71.4% of cases. Recurrence of VL occurred in nine patients and deaths were secondary to infectious complications (3/42 patients. Very low LTCD4 count (<100 cells/dL was the only independent variable associated with an unfavorable outcome in multivariate analysis (p = 0.03. CONCLUSION: Low LTCD4 count at presentation was associated with unfavorable outcome in VL/HIV patients.

  13. Evaluation of different commercial kits for HIV/HTLV-III EIA.

    Science.gov (United States)

    Rai, A; Kumari, S; Prabhakaran, P K

    1991-06-01

    Choice of an ideal, cost-effective and rapid diagnostic test for HIV infection is of immense value in developing countries like India where resources are limited. A number of commercial HIV antibody testing kits are now available with varying sensitivities and specificities. Six different commercial HIV kits namely, Wellcozyme, Flow HIV-TEKG, Abbott HIV EIA, Abbott VIA, Dip-stick EIA and Abbott env/core recombinant EIA were evaluated. Du-Pont Western blot (W.B.) kit was used as gold standard to compare the results. Of the 376 sera from various high-risk individuals screened, Wellcozyme kit yielded 100 per cent concordant results with W.B. Abbott VIA and Abbott env/core also yielded results in confirmation with W.B., excepting the fact that both detected one extra sample positive, which was negative in W.B. Abbott EIA yielded 4 false positive results. Dip-stick kit yielded the maximum number of false positives. The study indicated that 3 kits, namely Wellcozyme, Abbott VIA and Abbott EIA could be used to achieve optimum and acceptable results.

  14. Evaluating the Impact of Zimbabwe's Prevention of Mother-to-Child HIV Transmission Program: Population-Level Estimates of HIV-Free Infant Survival Pre-Option A.

    Directory of Open Access Journals (Sweden)

    Raluca Buzdugan

    Full Text Available We estimated HIV-free infant survival and mother-to-child HIV transmission (MTCT rates in Zimbabwe, some of the first community-based estimates from a UNAIDS priority country.In 2012 we surveyed mother-infant pairs residing in the catchment areas of 157 health facilities randomly selected from 5 of 10 provinces in Zimbabwe. Enrolled infants were born 9-18 months before the survey. We collected questionnaires, blood samples for HIV testing, and verbal autopsies for deceased mothers/infants. Estimates were assessed among i all HIV-exposed infants, as part of an impact evaluation of Option A of the 2010 WHO guidelines (rolled out in Zimbabwe in 2011, and ii the subgroup of infants unexposed to Option A. We compared province-level MTCT rates measured among women in the community with MTCT rates measured using program monitoring data from facilities serving those communities.Among 8568 women with known HIV serostatus, 1107 (12.9% were HIV-infected. Among all HIV-exposed infants, HIV-free infant survival was 90.9% (95% confidence interval (CI: 88.7-92.7 and MTCT was 8.8% (95% CI: 6.9-11.1. Sixty-six percent of HIV-exposed infants were still breastfeeding. Among the 762 infants born before Option A was implemented, 90.5% (95% CI: 88.1-92.5 were alive and HIV-uninfected at 9-18 months of age, and 9.1% (95%CI: 7.1-11.7 were HIV-infected. In four provinces, the community-based MTCT rate was higher than the facility-based MTCT rate. In Harare, the community and facility-based rates were 6.0% and 9.1%, respectively.By 2012 Zimbabwe had made substantial progress towards the elimination of MTCT. Our HIV-free infant survival and MTCT estimates capture HIV transmissions during pregnancy, delivery and breastfeeding regardless of whether or not mothers accessed health services. These estimates also provide a baseline against which to measure the impact of Option A guidelines (and subsequently Option B+.

  15. Evaluation of HIV, STI and CD4 results among voluntary attendees at the HIV/AIDS program of Mexico City

    Directory of Open Access Journals (Sweden)

    Luis Alfredo Juárez-Figueroa

    2017-03-01

    Full Text Available Objective. To describe results of HIV, sexually transmitted diseases (STI and CD4 counts at the HIV-specialized Condesa Clinic (CC in Mexico City. Materials and methods. Individuals who requested voluntary counseling and testing at CC were studied. We identified antibodies against HIV, syphilis, hepatitis C, and hepatitis B HBsAg. CD4 cell counts and viral load of HIV positive individuals were also obtained. Late HIV infection diagnosis was established if CD4 counts were lower than 200 cells/μL. Results. Global seroprevalence of HIV, syphilis, HBsAg, and anti HCV markers was of 20.1, 6, 1 and 1, respectively. Men displayed higher seroprevalence of infection markers than women. Among men, HIV infection was related to age and with all STI markers. Late HIV diagnosis was 31.8%. The risk of late HIV diagnosis was higher among women and it increased as age increased. Conclusions. Differences between genders regarding HIV and STIs prevalence as well as risk factors for HIV infection and late HIV diagnosis were observed.

  16. Combination Pod-Intravaginal Ring Delivers Antiretroviral Agents for HIV Prophylaxis: Pharmacokinetic Evaluation in an Ovine Model

    Science.gov (United States)

    Moss, John A.; Butkyavichene, Irina; Churchman, Scott A.; Gunawardana, Manjula; Fanter, Rob; Miller, Christine S.; Yang, Flora; Easley, Jeremiah T.; Marzinke, Mark A.; Hendrix, Craig W.; Smith, Thomas J.

    2016-01-01

    Preexposure prophylaxis (PrEP) against HIV using oral regimens based on the nucleoside reverse transcriptase inhibitor tenofovir disoproxil fumarate (TDF) has been effective to various degrees in multiple clinical trials, and the CCR5 receptor antagonist maraviroc (MVC) holds potential for complementary efficacy. The effectiveness of HIV PrEP is highly dependent on adherence. Incorporation of the TDF-MVC combination into intravaginal rings (IVRs) for sustained mucosal delivery could increase product adherence and efficacy compared with oral and vaginal gel formulations. A novel pod-IVR technology capable of delivering multiple drugs is described. The pharmacokinetics and preliminary local safety characteristics of a novel pod-IVR delivering a combination of TDF and MVC were evaluated in the ovine model. The device exhibited sustained release at controlled rates over the 28-day study and maintained steady-state drug levels in cervicovaginal fluids (CVFs). Dilution of CVFs during lavage sample collection was measured by ion chromatography using an inert tracer, allowing corrected drug concentrations to be measured for the first time. Median, steady-state drug levels in vaginal tissue homogenate were as follows: for tenofovir (TFV; in vivo hydrolysis product of TDF), 7.3 × 102 ng g−1 (interquartile range [IQR], 3.0 × 102, 4.0 × 103); for TFV diphosphate (TFV-DP; active metabolite of TFV), 1.8 × 104 fmol g−1 (IQR, 1.5 × 104, 4.8 × 104); and for MVC, 8.2 × 102 ng g−1 (IQR, 4.7 × 102, 2.0 × 103). No adverse events were observed. These findings, together with previous pod-IVR studies, have allowed several lead candidates to advance into clinical evaluation. PMID:27067321

  17. High-performance liquid chromatography assay for the quantification of HIV protease inhibitors and non-nucleoside reverse transcriptase inhibitors in human plasma.

    Science.gov (United States)

    Rezk, Naser L; Tidwell, Richard R; Kashuba, Angela D M

    2004-06-15

    An accurate, sensitive, and specific reverse-phase high-performance liquid chromatography (HPLC) assay for the simultaneous quantitative determination of HIV-protease inhibitors (PIs) (indinavir, IDV; amprenavir, APV; saquinavir, SQV; nelfinavir, NFV; ritonavir, RTV; and lopinavir, LPV) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) (nevirapine, NVP; delavirdine, DLV; and efavirenz, EFV) in human blood plasma is described. The method provides excellent resolution and peak shape for nine analytes through a linear gradient (36-86%) of 25% phosphate buffer (pH 4.5), 60% acetonitrile, 15% methanol, and 0.75 ml TFA, with a gradient mobile phase flow rate (0.9-1.1 ml) over 30 min run time. The optimized solid phase extraction (SPE) extraction method using (1.0 ml, 100mg BOND ELUT-C18 Varian) column provides a clean base line and high extraction efficiency using a 550 microl plasma sample. The method was validated over the range of 10-10,000 ng/ml for NVP, IDV, and SQV; 10-5000 ng/ml for EFV; 25-10000 ng/ml for APV; and 25-5000 ng/ml for DLV, NFV, RTV, and LPV. This method is accurate (average accuracies of three different concentrations ranged from 91 to 112%), and precise (within- and between-day precision measures ranged from 0.2 to 5.7% and 0.1 to 5.4%, respectively). This method is suitable for use in clinical pharmacokinetic studies as well as in therapeutic drug monitoring (TDM).

  18. ETV REPORT: EVALUATION OF HYDROMETRICS, INC., HIGH EFFICIENCY REVERSE OSMOSIS (HERO™) INDUSTRIAL WASTEWATER TREATMENT SYSTEM

    Science.gov (United States)

    Hydrometrics, founded in 1979 and located in Helena, MT, manufactures a commercial-ready High Efficiency Reverse Osmosis (HERO™) industrial wastewater treatment system. The system uses a three-stage reverse osmosis process to remove and concentrate metals for recovery while prod...

  19. One percent tenofovir applied topically to humanized BLT mice and used according to the CAPRISA 004 experimental design demonstrates partial protection from vaginal HIV infection, validating the BLT model for evaluation of new microbicide candidates.

    Science.gov (United States)

    Denton, Paul W; Othieno, Florence; Martinez-Torres, Francisco; Zou, Wei; Krisko, John F; Fleming, Elisa; Zein, Sima; Powell, Daniel A; Wahl, Angela; Kwak, Youn Tae; Welch, Brett D; Kay, Michael S; Payne, Deborah A; Gallay, Philippe; Appella, Ettore; Estes, Jacob D; Lu, Min; Garcia, J Victor

    2011-08-01

    Recent iPrEx clinical trial results provided evidence that systemic preexposure prophylaxis (PrEP) with emtricitabine (FTC) and tenofovir disoproxil fumarate (TDF) can partially prevent rectal HIV transmission in humans. Similarly, we have previously demonstrated that systemic administration of the same FTC-TDF combination efficiently prevented rectal transmission in humanized bone marrow/liver/thymus (BLT) mice. The CAPRISA 004 trial recently demonstrated that topical application of the tenofovir could partially prevent vaginal HIV-1 transmission in humans. To further validate the usefulness of the BLT mouse model for testing HIV prevention strategies, we evaluated the topical administration of tenofovir as used in CAPRISA 004 to prevent vaginal HIV transmission in BLT mice. Our results demonstrate that vaginally administered 1% tenofovir significantly reduced HIV transmission in BLT mice (P = 0.002). Together with the results obtained after systemic antiretroviral PrEP, these topical inhibitor data serve to validate the use of humanized BLT mice to evaluate both systemic and topical inhibitors of HIV transmission. Based on these observations, we tested six additional microbicide candidates for their ability to prevent vaginal HIV transmission: a C-peptide fusion inhibitor (C52L), a membrane-disrupting amphipathic peptide inhibitor (C5A), a trimeric d-peptide fusion inhibitor (PIE12-Trimer), a combination of reverse transcriptase inhibitors (FTC-TDF), a thioester zinc finger inhibitor (TC247), and a small-molecule Rac inhibitor (NSC23766). No protection was seen with the Rac inhibitor NSC23766. The thioester compound TC247 offered partial protection. Significant protection was afforded by FTC-TDF, and complete protection was offered by three different peptide inhibitors tested. Our results demonstrate that these effective topical inhibitors have excellent potential to prevent vaginal HIV transmission in humans.

  20. Real-world evaluation of the effectiveness of reversing camera and parking sensor technologies in preventing backover pedestrian injuries.

    Science.gov (United States)

    Keall, M D; Fildes, B; Newstead, S

    2017-02-01

    Backover injuries to pedestrians are a significant road safety issue, but their prevalence is underestimated as the majority of such injuries are often outside the scope of official road injury recording systems, which just focus on public roads. Based on experimental evidence, reversing cameras have been found to be effective in reducing the rate of collisions when reversing; the evidence for the effectiveness of reverse parking sensors has been mixed. The wide availability of these technologies in recent model vehicles provides impetus for real-world evaluations using crash data. A logistic model was fitted to data from crashes that occurred on public roads constituting 3172 pedestrian injuries in New Zealand and four Australian States to estimate the odds of backover injury (compared to other sorts of pedestrian injury crashes) for the different technology combinations fitted as standard equipment (both reversing cameras and sensors; just reversing cameras; just sensors; neither cameras nor sensors) controlling for vehicle type, jurisdiction, speed limit area and year of manufacture restricted to the range 2007-2013. Compared to vehicles without any of these technologies, reduced odds of backover injury were estimated for all three of these technology configurations: 0.59 (95% CI 0.39-0.88) for reversing cameras by themselves; 0.70 (95% CI 0.49-1.01) for both reversing cameras and sensors; 0.69 (95% CI 0.47-1.03) for reverse parking sensors by themselves. These findings are important as they are the first to our knowledge to present an assessment of real-world safety effectiveness of these technologies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Evaluation of EIA kits for the detection of HIV antibodies.

    Science.gov (United States)

    Ovcak, S; Drinovec, B; Likar, M

    1987-09-01

    We compared four commercially available enzyme immunoassay (EIA) kits for the detection of HIV antibodies using immunoblot analysis as a confirmatory test. The kits gave satisfactory results as far as sensitivity and specificity are concerned as required for the use in the laboratories of blood banks. For the sera of patients on haemodialysis, haemophiliacs, patients "under observation" for AIDS and homosexuals the results obtained by Behring and Organon kits were less satisfactory, as the number of false positive results was much higher than with kits produced by Genetic and Wellcome. The frequency of false negative results was small in the tests using Organon and Genetic kits, while using Behring and Wellcome kits no false negative results were found.

  2. [Application of different death evaluation indicators for HIV/AIDS prevention and treatment].

    Science.gov (United States)

    Zeng, L; Ma, Y

    2016-05-01

    AIDS has gradually changed from a fatal disease to a manageable chronic disease since the advent of antiretroviral drugs. In 2003, China initiated a national free antiretroviral treatment program for people living with HIV/AIDS, several death evaluation indicators have been used to evaluate public health effect of the program. Death evaluation indicators used frequently in domestic and overseas include mortality, case fatality rate, excess mortality, standard mortality ratio, years of potential life lost, disability-adjusted life year and life expectancy. This paper summarizes the different death indicators applied in effectiveness evaluation of HIV/AIDS prevention and treatment, elaborates the application range and significance of these indicators and suggests the research in related life expectancy and burden of disease which have not been conducted in China.

  3. Evaluation of a HIV voluntary opt-out screening program in a Singapore hospital.

    Science.gov (United States)

    Tan, Xin Quan; Goh, Wei-Ping; Venkatachalam, Indumathi; Goh, Diana; Sridhar, Revathi; Chan, Hwang Ching; Archuleta, Sophia

    2015-01-01

    Early diagnosis of human immunodeficiency virus (HIV) allows for appropriately timed interventions with improved outcomes, but HIV screening among asymptomatic persons and the general population in Singapore remains low. In 2008, Singapore's Ministry of Health implemented HIV voluntary opt-out screening (VOS) for hospitalised adults. We evaluated the outcome of VOS and surveyed reasons for its low uptake in our institution. We assessed the outcomes of the VOS programme from January 2010 to December 2013 at National University Hospital, a 1081-bed tertiary hospital in Singapore. We also examined reasons for opting-in and opting-out using an interviewer-administered structured questionnaire in a representative sample in January 2013. 107,523 patients fulfilled VOS criteria and were offered HIV screening, of which 5215 (4.9%) agreed to testing. 4850 (93.1%) of those who opted-in had an HIV test done. Three (0.06%) tested positive for HIV. 238 patients (14.2%) were surveyed regarding reasons for opting-in or out of VOS. 21 (8.8%) had opted-in. Patients who opted-in were likely to be younger, more educated and reported having more regular sexual partners. Type of housing, number of casual sexual partners, sexual orientation, intravenous drug use, condom use and previous sexually transmitted infection were not associated with deciding to opt-in/out. Patients' most common reasons for opting-out were: belief that they were at low risk (50.2%), belief that they were too old (26.8%), cost (6.9%) and aversion to venepuncture (6.5%). The most common reason for opting-in was desire to know their HIV status (47.6%). The success of an HIV-VOS program is largely determined by test uptake. Our study showed that the majority of eligible VOS patients opted-out of HIV screening. Given the considerable cost and low yield of this programme, more needs to be done to better equip patients in self-risk assessment and opting in to testing.

  4. Evaluation of a HIV voluntary opt-out screening program in a Singapore hospital.

    Directory of Open Access Journals (Sweden)

    Xin Quan Tan

    Full Text Available BACKGROUND: Early diagnosis of human immunodeficiency virus (HIV allows for appropriately timed interventions with improved outcomes, but HIV screening among asymptomatic persons and the general population in Singapore remains low. In 2008, Singapore's Ministry of Health implemented HIV voluntary opt-out screening (VOS for hospitalised adults. We evaluated the outcome of VOS and surveyed reasons for its low uptake in our institution. METHODS: We assessed the outcomes of the VOS programme from January 2010 to December 2013 at National University Hospital, a 1081-bed tertiary hospital in Singapore. We also examined reasons for opting-in and opting-out using an interviewer-administered structured questionnaire in a representative sample in January 2013. RESULTS: 107,523 patients fulfilled VOS criteria and were offered HIV screening, of which 5215 (4.9% agreed to testing. 4850 (93.1% of those who opted-in had an HIV test done. Three (0.06% tested positive for HIV. 238 patients (14.2% were surveyed regarding reasons for opting-in or out of VOS. 21 (8.8% had opted-in. Patients who opted-in were likely to be younger, more educated and reported having more regular sexual partners. Type of housing, number of casual sexual partners, sexual orientation, intravenous drug use, condom use and previous sexually transmitted infection were not associated with deciding to opt-in/out. Patients' most common reasons for opting-out were: belief that they were at low risk (50.2%, belief that they were too old (26.8%, cost (6.9% and aversion to venepuncture (6.5%. The most common reason for opting-in was desire to know their HIV status (47.6%. CONCLUSION: The success of an HIV-VOS program is largely determined by test uptake. Our study showed that the majority of eligible VOS patients opted-out of HIV screening. Given the considerable cost and low yield of this programme, more needs to be done to better equip patients in self-risk assessment and opting in to testing.

  5. Economic Evaluation of a Hybrid Desalination System Combining Forward and Reverse Osmosis

    Science.gov (United States)

    Choi, Yongjun; Cho, Hyeongrak; Shin, Yonghyun; Jang, Yongsun; Lee, Sangho

    2015-01-01

    This study seeks to evaluate the performance and economic feasibility of the forward osmosis (FO)–reverse osmosis (RO) hybrid process; to propose a guideline by which this hybrid process might be more price-competitive in the field. A solution-diffusion model modified with film theory was applied to analyze the effects of concentration polarization, water, and salt transport coefficient on flux, recovery, seawater concentration, and treated wastewater of the FO process of an FO-RO hybrid system. A simple cost model was applied to analyze the effects of flux; recovery of the FO process; energy; and membrane cost on the FO-RO hybrid process. The simulation results showed that the water transport coefficient and internal concentration polarization resistance are very important factors that affect performance in the FO process; however; the effect of the salt transport coefficient does not seem to be large. It was also found that the flux and recovery of the FO process, the FO membrane, and the electricity cost are very important factors that influence the water cost of an FO-RO hybrid system. This hybrid system can be price-competitive with RO systems when its recovery rate is very high, the flux and the membrane cost of the FO are similar to those of the RO, and the electricity cost is expensive. The most important thing in commercializing the FO process is enhancing performance (e.g.; flux and the recovery of FO membranes). PMID:26729176

  6. Evaluation of emerging contaminants in a drinking water treatment plant using electrodialysis reversal technology.

    Science.gov (United States)

    Gabarrón, S; Gernjak, W; Valero, F; Barceló, A; Petrovic, M; Rodríguez-Roda, I

    2016-05-15

    Emerging contaminants (EC) have gained much attention with globally increasing consumption and detection in aquatic ecosystems during the last two decades from ng/L to lower ug/L. The aim of this study was to evaluate the occurrence and removal of pharmaceutically active compounds (PhACs), endocrine disrupting chemicals (EDCs) and related compounds in a Drinking Water Treatment Plant (DWTP) treating raw water from the Mediterranean Llobregat River. The DWTP combined conventional treatment steps with the world's largest electrodialysis reversal (EDR) facility. 49 different PhACs, EDCs and related compounds were found above their limit of quantification in the influent of the DWTP, summing up to a total concentration of ECs between 1600-4200 ng/L. As expected, oxidation using chlorine dioxide and granular activated carbon filters were the most efficient technologies for EC removal. However, despite the low concentration detected in the influent of the EDR process, it was also possible to demonstrate that this process partially removed ionized compounds, thereby constituting an additional barrier against EC pollution in the product. In the product of the EDR system, only 18 out of 49 compounds were quantifiable in at least one of the four experimental campaigns, showing in all cases removals higher than 65% and often beyond 90% for the overall DWTP process. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Economic Evaluation of a Hybrid Desalination System Combining Forward and Reverse Osmosis

    Directory of Open Access Journals (Sweden)

    Yongjun Choi

    2015-12-01

    Full Text Available This study seeks to evaluate the performance and economic feasibility of the forward osmosis (FO–reverse osmosis (RO hybrid process; to propose a guideline by which this hybrid process might be more price-competitive in the field. A solution-diffusion model modified with film theory was applied to analyze the effects of concentration polarization, water, and salt transport coefficient on flux, recovery, seawater concentration, and treated wastewater of the FO process of an FO-RO hybrid system. A simple cost model was applied to analyze the effects of flux; recovery of the FO process; energy; and membrane cost on the FO-RO hybrid process. The simulation results showed that the water transport coefficient and internal concentration polarization resistance are very important factors that affect performance in the FO process; however; the effect of the salt transport coefficient does not seem to be large. It was also found that the flux and recovery of the FO process, the FO membrane, and the electricity cost are very important factors that influence the water cost of an FO-RO hybrid system. This hybrid system can be price-competitive with RO systems when its recovery rate is very high, the flux and the membrane cost of the FO are similar to those of the RO, and the electricity cost is expensive. The most important thing in commercializing the FO process is enhancing performance (e.g.; flux and the recovery of FO membranes.

  8. Economic Evaluation of a Hybrid Desalination System Combining Forward and Reverse Osmosis.

    Science.gov (United States)

    Choi, Yongjun; Cho, Hyeongrak; Shin, Yonghyun; Jang, Yongsun; Lee, Sangho

    2015-12-29

    This study seeks to evaluate the performance and economic feasibility of the forward osmosis (FO)-reverse osmosis (RO) hybrid process; to propose a guideline by which this hybrid process might be more price-competitive in the field. A solution-diffusion model modified with film theory was applied to analyze the effects of concentration polarization, water, and salt transport coefficient on flux, recovery, seawater concentration, and treated wastewater of the FO process of an FO-RO hybrid system. A simple cost model was applied to analyze the effects of flux; recovery of the FO process; energy; and membrane cost on the FO-RO hybrid process. The simulation results showed that the water transport coefficient and internal concentration polarization resistance are very important factors that affect performance in the FO process; however; the effect of the salt transport coefficient does not seem to be large. It was also found that the flux and recovery of the FO process, the FO membrane, and the electricity cost are very important factors that influence the water cost of an FO-RO hybrid system. This hybrid system can be price-competitive with RO systems when its recovery rate is very high, the flux and the membrane cost of the FO are similar to those of the RO, and the electricity cost is expensive. The most important thing in commercializing the FO process is enhancing performance (e.g.; flux and the recovery of FO membranes).

  9. Evaluation of reverse-transcriptase PCR as a diagnostic tool to confirm Japanese encephalitis virus infection.

    Science.gov (United States)

    Saxena, Vandana; Mishra, Virendra K; Dhole, Tapan N

    2009-04-01

    Japanese encephalitis (JE) is a serious central nervous system infection and major public health problem in several countries of Southeast Asia including India. This study evaluated the use of IgM ELISA and reverse-transcriptase (RT)-PCR in blood and cerebrospinal fluid (CSF) samples from acute encephalitis patients for the detection of Japanese encephalitis virus (JEV). Forty-four children suffering from acute encephalitis were enrolled, and 36 were selected from whom both CSF and serum samples were available. Twenty-two of the 36 CSF samples were positive for JEV by IgM ELISA and all were negative by RT-PCR. Twenty-three of the 36 serum samples were positive by IgM ELISA while 28 were positive by RT-PCR. Total positivity for JEV infection in CSF and serum samples was 66.7% (24/36) and 83.3% (30/36) respectively by one or both tests. The overall positivity for JEV infection was 86.1% (31/36). We suggest that the use of RT-PCR in serum samples during the early days of JEV infection may be helpful in confirming diagnosis in those cases which are negative for JEV-specific IgM antibodies in both serum and CSF samples.

  10. A comparison of the ability of rilpivirine (TMC278 and selected analogues to inhibit clinically relevant HIV-1 reverse transcriptase mutants

    Directory of Open Access Journals (Sweden)

    Johnson Barry C

    2012-12-01

    Full Text Available Abstract Background The recently approved anti-AIDS drug rilpivirine (TMC278, Edurant is a nonnucleoside inhibitor (NNRTI that binds to reverse transcriptase (RT and allosterically blocks the chemical step of DNA synthesis. In contrast to earlier NNRTIs, rilpivirine retains potency against well-characterized, clinically relevant RT mutants. Many structural analogues of rilpivirine are described in the patent literature, but detailed analyses of their antiviral activities have not been published. This work addresses the ability of several of these analogues to inhibit the replication of wild-type (WT and drug-resistant HIV-1. Results We used a combination of structure activity relationships and X-ray crystallography to examine NNRTIs that are structurally related to rilpivirine to determine their ability to inhibit WT RT and several clinically relevant RT mutants. Several analogues showed broad activity with only modest losses of potency when challenged with drug-resistant viruses. Structural analyses (crystallography or modeling of several analogues whose potencies were reduced by RT mutations provide insight into why these compounds were less effective. Conclusions Subtle variations between compounds can lead to profound differences in their activities and resistance profiles. Compounds with larger substitutions replacing the pyrimidine and benzonitrile groups of rilpivirine, which reorient pocket residues, tend to lose more activity against the mutants we tested. These results provide a deeper understanding of how rilpivirine and related compounds interact with the NNRTI binding pocket and should facilitate development of novel inhibitors.

  11. A Novel Lectin with Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Dried Fruiting Bodies of the Monkey Head Mushroom Hericium erinaceum

    Science.gov (United States)

    Li, Yanrui; Zhang, Guoqing; Ng, Tzi Bun; Wang, Hexiang

    2010-01-01

    A lectin designated as Hericium erinaceum agglutinin (HEA) was isolated from dried fruiting bodies of the mushroom Hericium erinaceum with a chromatographic procedure which entailed DEAE-cellulose, CM-cellulose, Q-Sepharose, and FPLC Superdex 75. Its molecular mass was estimated to be 51 kDa and its N-terminal amino acid sequences was distinctly different from those of other isolated mushroom lectins. The hemagglutinating activity of HEA was inhibited at the minimum concentration of 12.5 mM by inulin. The lectin was stable at pH 1.9–12.1 and at temperatures up to 70°C, but was inhibited by Hg2+, Cu2+, and Fe3+ ions. The lectin exhibited potent mitogenic activity toward mouse splenocytes, and demonstrated antiproliferative activity toward hepatoma (HepG2) and breast cancer (MCF7) cells with an IC50 of 56.1 μM and 76.5 μM, respectively. It manifested HIV-1 reverse transcriptase inhibitory activity with an IC50 of 31.7 μM. The lectin exhibited potent mitogenic activity toward murine splenocytes but was devoid of antifungal activity. PMID:20625408

  12. A Novel Lectin with Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Dried Fruiting Bodies of the Monkey Head Mushroom Hericium erinaceum

    Directory of Open Access Journals (Sweden)

    Yanrui Li

    2010-01-01

    Full Text Available A lectin designated as Hericium erinaceum agglutinin (HEA was isolated from dried fruiting bodies of the mushroom Hericium erinaceum with a chromatographic procedure which entailed DEAE-cellulose, CM-cellulose, Q-Sepharose, and FPLC Superdex 75. Its molecular mass was estimated to be 51 kDa and its N-terminal amino acid sequences was distinctly different from those of other isolated mushroom lectins. The hemagglutinating activity of HEA was inhibited at the minimum concentration of 12.5 mM by inulin. The lectin was stable at pH 1.9–12.1 and at temperatures up to 70∘C, but was inhibited by Hg2+, Cu2+, and Fe3+ ions. The lectin exhibited potent mitogenic activity toward mouse splenocytes, and demonstrated antiproliferative activity toward hepatoma (HepG2 and breast cancer (MCF7 cells with an IC50 of 56.1 M and 76.5 M, respectively. It manifested HIV-1 reverse transcriptase inhibitory activity with an IC50 of 31.7 M. The lectin exhibited potent mitogenic activity toward murine splenocytes but was devoid of antifungal activity.

  13. Isolation and characterization of a French bean hemagglutinin with antitumor, antifungal, and anti-HIV-1 reverse transcriptase activities and an exceptionally high yield.

    Science.gov (United States)

    Lam, S K; Ng, T B

    2010-05-01

    A dimeric 64-kDa hemagglutinin was isolated with a high yield from dried Phaseolus vulgaris cultivar "French bean number 35" seeds using a chromatographic protocol that involved Blue-Sepharose, Q-Sepharose, and Superdex 75. The yield was exceptionally high (1.1g hemagglutinin per 100g seed), which is around 10-85 times higher than other Phaseolus cultivars. Its N-terminal sequence resembled those of other Phaseolus hemagglutinins. The hemagglutinating activity of the hemagglutinin was stable in the pH range 6-8, and in the temperature range 0 degrees C-50 degrees C. It inhibited HIV-1 reverse transcriptase with an IC50 of 2microM. It suppressed mycelial growth in Valsa mali with an IC50 of 10microM. It inhibited proliferation of hepatoma HepG2 cells and breast cancer MCF-7 cells with an IC50 of 100 and 2microM, respectively. It had no antiproliferative effect on normal embryonic liver WRL68 cells. Copyright 2009 Elsevier GmbH. All rights reserved.

  14. Enzymatic properties and sensitivity to inhibitors of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase with Glu-138-->Arg and Tyr-188-->His mutations.

    Science.gov (United States)

    Zhang, H; Vrang, L; Bäckbro, K; Unge, T; Noréen, R; Oberg, B

    1994-05-01

    Two mutants of HIV-1 reverse transcriptase (RT), Tyr-188-->His and Glu-138-->Arg have been prepared and their catalytic properties and sensitivities to inhibitors studied. As compared to wild type RT, a reduction in catalytic efficiency and turn over number was observed, especially for the Tyr-188-->His mutant. The non-nucleoside inhibitors nevirapine, L-697,661 and 9-Cl-TIBO caused a mixed type of inhibition of RT (Arg-138) with respect to substrate, and with the exception of a non-competitive inhibition by nevirapine, also a mixed type of inhibition of RT (His-188). Foscarnet (PFA) caused a non-competitive type of inhibition of RT (Arg-138) and a mixed inhibition of RT (His-188). The inhibition by ddG-TP was competitive with both mutant RTs. Inhibition by nevirapine gave IC50 values of 0.15, 0.23 and 0.72 microM; by 9-Cl-TIBO of 0.20, 2.50 and 10.3 microM; by L-697,661 of 0.064, 0.28 and 0.60 microM; by ddGTP of 0.13, 0.14 and 0.02 microM; by PFA of 17.0, 48.0 and 15.0 microM for RT wt, RT (Arg-138) and RT (His-188), respectively.

  15. Fusion to the Lysosome Targeting Signal of the Invariant Chain Alters the Processing and Enhances the Immunogenicity of HIV-1 Reverse Transcriptase.

    Science.gov (United States)

    Starodubova, E S; Isaguliants, M G; Kuzmenko, Y V; Latanova, A A; Krotova, O A; Karpov, V L

    2014-01-01

    Intracellular processing of the antigen encoded by a DNA vaccine is one of the key steps in generating an immune response. Immunization with DNA constructs targeted to the endosomal-lysosomal compartments and to the MHC class II pathway can elicit a strong immune response. Herein, the weakly immunogenic reverse transcriptase of HIV-1 was fused to the minimal lysosomal targeting motif of the human MHC class II invariant chain. The motif fused to the N-terminus shifted the enzyme intracellular localization and accelerated its degradation. Degradation of the chimeric protein occurred predominantly in the lysosomal compartment. BALB/c mice immunized with the plasmid encoding the chimeric protein demonstrated an enhanced immune response, in the form of an increased antigen-specific production of Th1 cytokines, INF-γ and IL-2, by mouse splenocytes. Moreover, the majority of the splenocytes secreted both cytokines; i.e., were polyfunctional. These findings suggest that retargeting of the antigen to the lysosomes enhances the immune response to DNA vaccine candidates with low intrinsic immunogenicity.

  16. Purification and Characterization of a White Laccase with Pronounced Dye Decolorizing Ability and HIV-1 Reverse Transcriptase Inhibitory Activity from Lepista nuda

    Directory of Open Access Journals (Sweden)

    Mengjuan Zhu

    2016-03-01

    Full Text Available A strain LN07 with high laccase yield was identified as basidiomycete fungus Lepista nuda from which a white laccase without type I copper was purified and characterized. The laccase was a monomeric protein with a molecular mass of 56 kDa. Its N-terminal amino acid sequence was AIGPAADLHIVNKDISPDGF. Besides, eight inner peptide sequences were determined and lac4, lac5 and lac6 sequences were in the Cu2+ combination and conservation zones of laccases. HIV-1 reverse transcriptase was inhibited by the laccase with a half-inhibitory concentration of 0.65 μM. Cu2+ ions (1.5 mM enhanced the laccase production and the optimal pH and temperature of the laccase were pH 3.0 and 50 °C, respectively. The Km and Vmax of the laccase using ABTS as substrate were respectively 0.19 mM and 195 μM. Several dyes including laboratory dyes and textile dyes used in this study, such as Methyl red, Coomassie brilliant blue, Reactive brilliant blue and so on, were decolorized in different degrees by the purified laccase. By LC-MS analysis, Methyl red was structurally degraded by the laccase. Moreover, the laccase affected the absorbance at the maximum wavelength of many pesticides. Thus, the white laccase had potential commercial value for textile finishing and wastewater treatment.

  17. Comparative Molecular Field Analysis (CoMFA for Thiotetrazole Alkynylacetanilides, a Non-Nucleoside Inhibitor of HIV-1 Double Mutant K103N/Y181C Reverse Transcriptase

    Directory of Open Access Journals (Sweden)

    Sivan Sree Kanth

    2009-01-01

    Full Text Available Non-nucleoside reverse transcriptase inhibitors (NNRTIs are allosteric inhibitors of the HIV-1 reverse transcriptase. HIV-1 resistance to nucleoside RT inhibitors such as AZT can arise through mutations in the coding region of RT. Recently a series of thioterazolyl acetanilides were reported as potent inhibitors of HIV-1 wild type and double mutant K103N/Y181C reverse transcriptase. In the present study 3D quantitative structure activity relationship (3D QSAR studies involving comparative molecular field analysis (CoFMA were performed on 28 thiotetrazole alkynylacetanilides. CoMFA was performed using sibyl software 6.7v. The molecules were divided into training set and test set. The developed model based on training set containing 20 molecules gave Q2 value of 0.587 and non cross validation R2 value of 0.961 and standard error of estimate 0.098 for CoMFA. The steric and electrostatic contributions are 42.7% and 57.3% The cross validation on test set gave R2 value of 0.74. The CoMFA model provided the most significant correlation of steric and electrostatic fields with biological activities. The information rendered by 3D QSAR model may afford valuable clues to optimize the lead and design new potential inhibitors

  18. Palbociclib, a selective inhibitor of cyclin-dependent kinase4/6, blocks HIV-1 reverse transcription through the control of sterile α motif and HD domain-containing protein-1 (SAMHD1) activity.

    Science.gov (United States)

    Pauls, Eduardo; Badia, Roger; Torres-Torronteras, Javier; Ruiz, Alba; Permanyer, Marc; Riveira-Muñoz, Eva; Clotet, Bonaventura; Marti, Ramón; Ballana, Ester; Esté, José A

    2014-09-24

    Sterile α motif and HD domain-containing protein-1 (SAMHD1) inhibits HIV-1 reverse transcription by decreasing the pool of intracellular deoxynucleotides. SAMHD1 is controlled by cyclin-dependent kinase (CDK)-mediated phosphorylation. However, the exact mechanism of SAMHD1 regulation in primary cells is unclear. We explore the effect of palbociclib, a CDK6 inhibitor, in HIV-1 replication. Human primary monocytes were differentiated into macrophages with monocyte-colony stimulating factor and CD4 T lymphocytes stimulated with phytohaemagglutinin (PHA)/interleukin-2. Cells were treated with palbociclib and then infected with a Green fluorescent protein-expressing HIV-1 or R5 HIV-1 BaL. Viral DNA was measured by quantitative PCR and infection assessed by flow cytometry. Deoxynucleotide triphosphate (dNTP) content was determined using a polymerase-based method. Pan-CDK inhibitors AT7519, roscovitine and purvalanol A reduced SAMHD1 phosphorylation. HIV-1 replication was blocked by AT7519 (66.4 ± 3.8%; n = 4), roscovitine (47.3 ± 3.9%; n = 4) and purvalanol A (55.7 ± 15.7%; n = 4) at subtoxic concentrations. Palbociclib, a potent and selective CDK6 inhibitor, blocked SAMHD1 phosphorylation, intracellular dNTP levels, HIV-1 reverse transcription and HIV-1 replication in primary macrophages and CD4 T lymphocytes. Notably, treatment of macrophages with palbociclib led to reduced CDK2 activation, measured as the phosphorylation of the T-loop at the Thr160. The antiviral effect was lost when SAMHD1 was degraded by Vpx, providing further evidence for a role of SAMHD1 in mediating the antiretroviral effect. Our results indicate that SAMHD1-mediated HIV-1 restriction is controlled by CDK as previously suggested but point to a preferential role for CDK2 and CDK6 as mediators of SAMHD1 activation. Our study provides a new signaling pathway susceptible for the development of new therapeutic approaches against HIV-1 infection.

  19. Community-based interventions that work to reduce HIV stigma and discrimination: results of an evaluation study in Thailand.

    Science.gov (United States)

    Jain, Aparna; Nuankaew, Ratana; Mongkholwiboolphol, Nungruthai; Banpabuth, Arunee; Tuvinun, Rachada; Oranop Na Ayuthaya, Pakprim; Richter, Kerry

    2013-11-13

    HIV stigma and discrimination are major issues affecting people living with HIV in their everyday lives. In Thailand, a project was implemented to address HIV stigma and discrimination within communities with four activities: (1) monthly banking days; (2) HIV campaigns; (3) information, education and communication (IEC) materials and (4) "Funfairs." This study evaluates the effect of project interventions on reducing community-level HIV stigma. A repeated cross-sectional design was developed to measure changes in HIV knowledge and HIV-related stigma domains among community members exposed to the project. Two cross-sectional surveys were implemented at baseline (respondent n=560) and endline (respondent n=560). T-tests were employed to assess changes on three stigma domains: fear of HIV infection through daily activity, shame associated with having HIV and blame towards people with HIV. Baseline scales were confirmed at endline, and each scale was regressed on demographic characteristics, HIV knowledge and exposure to intervention activities. No differences were observed in respondent characteristics at baseline and endline. Significant changes were observed in HIV transmission knowledge, fear of HIV infection and shame associated with having HIV from baseline to endline. Respondents exposed to three specific activities (monthly campaign, Funfair and IEC materials) were less likely to exhibit stigma along the dimensions of fear (3.8 points lower on average compared to respondents exposed to none or only one intervention; 95% CI: -7.3 to -0.3) and shame (4.1 points lower; 95% CI: -7.7 to -0.6), net of demographic controls and baseline levels of stigma. Personally knowing someone with HIV was associated with low fear and shame, and females were less likely to possess attitudes of shame compared to males. The multivariate linear models suggest that a combination of three interventions was critical in shifting community-level stigma--monthly campaign, Funfair and IEC

  20. Rationale, Design, and Baseline Characteristics of Beijing Prediabetes Reversion Program: A Randomized Controlled Clinical Trial to Evaluate the Efficacy of Lifestyle Intervention and/or Pioglitazone in Reversion to Normal Glucose Tolerance in Prediabetes

    National Research Council Canada - National Science Library

    Yingying Luo; Sanjoy K. Paul; Xianghai Zhou; Cuiqing Chang; Wei Chen; Xiaohui Guo; Jinkui Yang; Linong Ji; Hongyuan Wang

    2017-01-01

    .... Beijing Prediabetes Reversion Program (BPRP) would evaluate whether intensive lifestyle modification and/or pioglitazone could revert prediabetic state to normoglycemia and improve the risk factors of CVD as well. Methods...

  1. Evaluation of the HIV lay counselling and testing profession in South Africa.

    Science.gov (United States)

    Mwisongo, Aziza; Mehlomakhulu, Vuyelwa; Mohlabane, Neo; Peltzer, Karl; Mthembu, Jacque; Van Rooyen, Heidi

    2015-07-22

    With the launch of the national HIV Counselling and Testing (HCT) campaign in South Africa (SA), lay HIV counsellors, who had been trained in blood withdrawal, have taken up the role of HIV testing. This study evaluated the experiences, training, motivation, support, supervision, and workload of HIV lay counsellors and testers in South Africa. The aim was to identify gaps in their resources, training, supervision, motivation, and workload related to HCT services. In addition it explored their experiences with providing HIV testing under the task shifting context. The study was conducted in eight of South Africa's nine provinces. 32 lay counsellors were recruited from 67 HCT sites, and were interviewed using two questionnaires that included structured and semi-structured questions. One questionnaire focused on their role as HIV counsellors and the other on their role as HIV testers. Ninety-seven percent of counsellors reported that they have received training in counselling and testing. Many rated their training as more than adequate or adequate, with 15.6% rating it as not adequate. Respondents reported a lack of standardised counselling and testing training, and revealed gaps in counselling skills for specific groups such as discordant couples, homosexuals, older clients and children. They indicated health system barriers, including inadequate designated space for counselling, which compromises privacy and confidentiality. Lay counsellors carry the burden of counselling and testing nationally, and have other tasks such as administration and auxiliary duties due to staff shortages. This study demonstrates that HCT counselling and testing services in South Africa are mainly performed by lay counsellors and testers. They are challenged by inadequate work space, limited counselling skills for specific groups, a lack of standardised training policies and considerable administrative and auxiliary duties. To improve HCT services, there needs to be training needs with a

  2. Comparative evaluation of glasses reprocessing and reversible conditioning of calcinates; Evaluation comparative de la reprise des verres et du conditionnement reversible des calcinats

    Energy Technology Data Exchange (ETDEWEB)

    Boen, R

    2000-01-15

    Fission products and minor actinides separated during the spent fuel reprocessing treatment are industrially vitrified on-line and thus confined inside a glass matrix with admittedly durability properties. In the framework of the feasibility of a reversible conditioning, this document examines first the possible alternative ways of conditioning and storage of calcinates before vitrification, which may simplify the reversibility aspect. Such a conditioning must be compatible with the storage process, with a possible extraction of actinides and long-lived fission products, and with the vitrification process if no extraction is performed. Calcinates are pulverulent and comprise an important soluble fraction, a proportion of nitrates of about 30%, and release a high thermal power (17 kW/m{sup 3}) combined to a low thermal conductivity (0.1 to 0.15 W.m{sup -1} k{sup -1}). Among the different foreseeable solutions (denitration, mixing with another material, with or without compacting, dissolution inside another material..), the dissolution inside a borate seems to be the most acceptable with respect to the safety, feasibility and vitrification aspects. The thermal aspect of the storage remains complex as a specific container is necessary. In a second part, this report analyzes the possibility to re-extract back the long-lived radionuclides from vitrified wastes. The different possible ways to destroy the glass structure and to transfer the fission products and minor actinides in an aqueous solution compatible with an hydrometallurgical separation process are explored. Two processes are foreseeable: a low temperature dissolution process which requires a preliminary crushing and the handling of huge amounts of acids, and a both high and low temperature process which comprises the following steps: melting, fractionation by water tempering, addition of Na{sub 2}O or sodium tetraborate to make it sensible to hot leaching, separation of fission products and minor actinides

  3. The relationship between economic evaluations and HIV and AIDS treatment policies.

    Science.gov (United States)

    Taleski, Sarah Jane; Ahmed, Khaled; Whiteside, Alan

    2010-05-01

    Economics, and specifically economic evaluations, are increasingly being utilized to provide treatment policy guidance to decision makers. This article reviews work that has contributed to understanding of the relationship. There is a paucity of research explicitly investigating the association between economic evaluations and HIV and AIDS treatment policy. Where it does exist, it is weak. Factors contributing to the limited impact include lack of reliable and trusted data; absence of local cost-effectiveness data for different interventions; contradictory results; challenges associated with understanding complex economic/mathematical models; inefficient implementation of HIV and AIDS policies; inability to pursue long-term health planning needs; and political will. Consideration of the ways in which economic evaluations can have greater influence over HIV and AIDS policies is needed. The weak relationship between the two reflects the complicated and multifaceted decision-making process that is often influenced by socioeconomic and political factors. If an economic evaluation is to influence policy, then cognizance of this is important. Extending the economic toolkit to include broader-based models that incorporate political economy variables, but do not compromise on comprehension, validity and robustness, will offer better informed policy recommendations.

  4. Moving from the HIV Organ Policy Equity Act to HIV Organ Policy Equity in action: changing practice and challenging stigma.

    Science.gov (United States)

    Doby, Brianna L; Tobian, Aaron A R; Segev, Dorry L; Durand, Christine M

    2018-02-09

    The HIV Organ Policy Equity (HOPE) Act, signed in 2013, reversed the federal ban on HIV-to-HIV transplantation. In this review, we examine the progress in HOPE implementation, the current status of HIV-to-HIV transplantation, and remaining challenges. Pursuant to the HOPE Act, the Department of Health and Human Services revised federal regulations to allow HIV-to-HIV transplants under research protocols adherent to criteria published by the National Institutes of Health. The first HIV-to-HIV kidney and liver transplants were performed at Johns Hopkins in March of 2016. Legal and practical challenges remain. Further efforts are needed to educate potential HIV+ donors and to support Organ Procurement Organizations. As of November 2017, there are 22 transplant centers approved to perform HIV-to-HIV transplants in 10 United Network for Organ Sharing regions. To date, 16 Organ Procurement Organizations in 22 states have evaluated HIV+ donors. The National Institutes of Health-funded HOPE in Action: A Multicenter Clinical Trial of HIV-to-HIV Deceased Donor (HIVDD) Kidney Transplantation Kidney Trial will launch at 19 transplant centers in December of 2017. A HOPE in Action Multicenter HIVDD Liver Trial is in development. Significant progress toward full HOPE implementation has been made though barriers remain. Some challenges are unique to HIV-HIV transplantation, whereas others are amplifications of issues across the current transplant system. In addition to a public health benefit for all transplant candidates in the United States, partnership on the HOPE Act has the potential to address systemic challenges to national donation and transplantation.

  5. Effectiveness of antihypertensive therapy in HIV-positive patients: evaluation to 144 weeks

    Directory of Open Access Journals (Sweden)

    K Falasca

    2012-11-01

    Full Text Available Hypertension is more prevalent among HIV-infected subjects than in the general population, contributing to increased cardiovascular risk in HIV+ patients. The angiotensin II receptor blocker telmisartan is also a partial peroxisome proliferator activated receptor (PPAR-γ agonist, with documented effects on improving hypertension, lipid metabolism and renal function. Therefore telmisartan was found to be the first choice for treatment of HIV+ hypertensive patients. Aim of this study was to evaluate the durability on 144 weeks of treatment with telmisartan in HIV+ patients. 13 HIV+ Caucasian male patients treated with combined antiretroviral therapy (cART and discovered to be naïve hypertensive, were given 80 mg telmisartan daily. Systolic (SBP and diastolic (DBP blood pressure, viro-immunological, lipid and metabolic parameters, including triglycerides, cholesterol, insulin resistance (HOMA-IR, inflammatory markers, C-reactive protein (CRP, erythrocyte sedimentation rate (ESR, indexes of renal function and cardiovascular risk, microalbuminuria, cystatin C, were measured at baseline (T0, and after 24 (T24, 48 (T48, 72 (T72, 96 (T96, 120 (T120 and 144 (T144 weeks. Treatment with telmisartan decreased SBP and DBP levels during the 144 weeks of observation. We also observed improved HDL-cholesterol, HOMA-IR, microalbuminuria and cystatin C at the end of study. Triglycerides and total cholesterol significantly decreased and HDL-cholesterol significantly increased. Total cholesterol/HDL cholesterol ratio improved significantly. Throughout in the course of the trial our patients showed a significant improvement of the percentage of CD4+ and CD8+. Eventually in all 144 weeks of therapy with telmisartan 80 mg/day have not observed adverse events or dropouts. Telmisartan was effective in improving hypertension, lipid metabolism and renal function in 144 weeks of evaluation. It determines the improvement of cystatin C and microalbuminuria, markers of

  6. HIV prevention outreach in commercial gay venues in large cities: evaluation findings from London.

    Science.gov (United States)

    Bonell, Chris; Strange, V; Allen, E; Barnett-Page, E

    2006-08-01

    Human immunodeficiency virus (HIV) prevention delivered in gay venues in US cities has been found to be effective in reducing HIV transmission in the 1990s but effects might not be generalizable to different times and settings. Doubts have been raised about: outreach's ability to address skills and explore personal behaviour; big-city commercial gay venues being appropriate sites for outreach because of gossip and social surveillance; and acceptability of outreach by professionals rather than 'popular opinion formers'. We evaluated coverage, feasibility, acceptability and perceived impact of venue-based HIV prevention outreach by professionals in London, employing observation, surveys and interviews with venue-users, and focus groups/semi-structured interviews with workers. We found high coverage especially among target groups. Addressing negotiation skills and personal behaviour was feasible but required worker motivation and skill. Social surveillance rarely impeded work. Gay men generally found outreach acceptable and useful, and professionals were not regarded negatively. Impact on knowledge was commonly reported; impacts on negotiation skills and reflection on personal behaviour were more common among men experiencing longer contacts. In conclusion, professional HIV prevention outreach in gay venues in large cities is a feasible and acceptable intervention with significant potential impacts. Workers need to be well briefed and trained to maximize impact.

  7. An improved microtiter assay for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma

    Directory of Open Access Journals (Sweden)

    Chen Yunyun

    2003-12-01

    Full Text Available Abstract Background The anti-HIV-1 neutralizing antibody assay is widely used in AIDS vaccine research and other experimental and clinical studies. The vital dye staining method applied in the detection of anti-HIV-1 neutralizing antibody has been used in many laboratories. However, the unknown factor(s in sera or plasma affected cell growth and caused protection when the tested sera or plasma was continuously maintained in cell culture. In addition, the poor solubility of neutral red in medium (such as RPMI-1640 also limited the use of this assay. Methods In this study, human T cell line C8166 was used as host cells, and 3-(4,5-Dimethyl-2-thiazolyl-2,5-diphenyl-2H-tetrazolium bromide (MTT instead of neutral red was used as vital dye. In order to avoid the effect of the unknown factor(s, the tested sera or plasma was removed by a washout procedure after initial 3–6 h culture in the assay. Result This new assay eliminated the effect of the tested sera or plasma on cell growth, improved the reliability of detection of anti-HIV-1 neutralizing antibody, and showed excellent agreement with the p24 antigen method. Conclusion The results suggest that the improved assay is relatively simple, highly duplicable, cost-effective, and well reliable for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma.

  8. An evaluation of an education programme on HIV infection using puppetry and street theatre.

    Science.gov (United States)

    Skinner, D; Metcalf, C A; Seager, J R; de Swardt, J S; Laubscher, J A

    1991-01-01

    'Puppets Against AIDS' is a novel educational medium being used to try to reduce the spread of HIV infection in South Africa. It involves the use of street theatre employing two-metre-high puppets who act out a story of how one person, who is infected with HIV, passes it onto a series of other people until he eventually dies. The puppet show was evaluated in two phases. The first involved a content analysis of a video recording of the show by a multidisciplinary group, according to a set of criteria for appropriate education on HIV infection. This show was found to be professional and comprehensive in terms of the educational messages provided. Some suggestions were made for improvements. The second phase was a before and after study of the impact on the audience at a series of live shows. The show made a significant contribution to knowledge and intended behaviour in the short term. Overall it was felt that the show does make a valuable contribution, but could be made more effective if incorporated into existing community-based education programmes on HIV infection.

  9. Prothrombin Time Tests for the Monitoring of Direct Oral Anticoagulants and Their Evaluation as Indicators of the Reversal Effect.

    Science.gov (United States)

    Nagakari, Kunihiko; Emmi, Mari; Iba, Toshiaki

    2017-09-01

    The prompt assessment and the reversal of direct oral anticoagulants (DOACs) are urgent matters in the emergency care setting. Thus, we planned to elucidate the adequate prothrombin time (PT) test for the evaluation of the anticoagulant effects of various DOACs. The anticoagulant effects of rivaroxaban, apixaban, and edoxaban were measured with 3 PT tests (Triniclot PT Excel S, Neoplastin R, and Thromborel S). Human plasma was spiked with each DOAC at a range of 0 to 1000 ng/mL, and the PT was measured using each PT test. In another series, the reversal effect of either 4-factor prothrombin complex concentrate (PCC) or activated PCC (aPCC) was evaluated with each PT test. All PT reagents correlated with the concentrations of each DOAC, however, the reactivity was considerably different between the DOACs and the PT tests. A prolonged PT with DOACs was reversed both by PCC and aPCC in a dose-dependent manner; however, Triniclot PT Excel S showed reprolongation of the PT with a higher dose of PCC. The proper choice of PT test is necessary for the assessments of the anticoagulant activity of DOACs. It is also important to understand the different characteristics of each PT test for the assessment of the reversal effects of PCC.

  10. [Treatment of proximal humeral fractures by reverse shoulder arthroplasty: mid-term evaluation of functional results and Notching].

    Science.gov (United States)

    Hernández-Elena, J; de la Red-Gallego, M Á; Garcés-Zarzalejo, C; Pascual-Carra, M A; Pérez-Aguilar, M D; Rodríguez-López, T; Alfonso-Fernández, A; Pérez-Núñez, M I

    2015-01-01

    An analysis was made on relationship between Notching and functional and radiographic parameters after treatment of acute proximal humeral fractures with reverse total shoulder arthroplasty. A retrospective evaluation was performed on 37 patients with acute proximal humeral fracture treated by reversed shoulder arthroplasty. The mean follow-up was 24 months. Range of motion, intraoperative and postoperative complications were recorded. Nerot's classification was used to evaluate Notching. Patient satisfaction was evaluated with the Constant Score (CS). Statistical analysis was performed to evaluate the relationship between Notching and glenosphere position, or functional outcomes. Mean range of elevation, abduction, external and internal rotation were 106.22°, 104.46°, 46.08° and 40.27°, respectively. Mean CS was 63. Notching was present at 12 months in 29% of patients. Statistical analysis showed significance differences between age and CS, age and notching development, and tilt with notching. No statistical significance differences were found between elevation, abduction, internal and external rotation and CS either with scapular or glenosphere-neck angle. Reverse shoulder arthroplasty is a valuable option for acute humeral fractures in patients with osteoporosis and cuff-tear arthropathy. It leads to early pain relief and shoulder motion. Nevertheless, it is not exempt from complications, and long-term studies are needed to determine the importance of notching. Copyright © 2014 SECOT. Published by Elsevier Espana. All rights reserved.

  11. Non Castigat Ridendo Mores: evaluating the effectiveness of humor appeal in printed advertisements for HIV/AIDS prevention in Italy.

    Science.gov (United States)

    Soscia, Isabella; Turrini, Alex; Tanzi, Emilio

    2012-01-01

    This article investigates the effects of different emotional appeals in HIV/AIDS prevention campaigns using printed advertisements. More specifically, it examines the effectiveness of humor appeal compared with shock and fear appeals. The authors experimentally test the level of attention drawn and the spontaneous recall arising when young Italian adults are shown different HIV/AIDS prevention campaigns. Findings show that humor appeals are less effective than fear and shock appeals, evidencing the failures in HIV/AIDS prevention campaigns in Italy, a country where the former communication strategy has been used in substantive ways. The results also indicate the higher effectiveness of fear appeals (over shock and humor) in printed HIV/AIDS advertising campaigns. The implications of these results for further studies and for improving the design, implementation, and evaluation of HIV/AIDS campaign efforts are also discussed.

  12. Mild desalination demo pilot: New normalization approach to effectively evaluate electrodialysis reversal technology

    Directory of Open Access Journals (Sweden)

    Roel Bisselink

    2016-06-01

    Full Text Available Key performance indicators for characterization of nanofiltration performance are well developed, similar key performance indicators for electrodialysis reversal are however underdeveloped. Under the E4Water project Dow Benelux BV and Evides Industriewater BV operate a pilot facility to compare both technologies for their application to mildly desalinate a variety of brackish water streams. Normalized pressure drop, normalized current efficiency and normalized membrane resistance proved to be a useful tool to interpret process performance and to initiate a cleaning procedure if required. The availability of these normalized key performance indicators enables optimization and process monitoring and control of electrodialysis reversal independent of the continuously changing conditions of the feed water.

  13. Tapping youth as agents for change: evaluation of a peer leadership HIV/AIDS intervention.

    Science.gov (United States)

    Pearlman, Deborah N; Camberg, Lois; Wallace, Laurie Jo; Symons, Paul; Finison, Lorenz

    2002-07-01

    To evaluate the impact of a community-based HIV/AIDS peer leadership prevention program on newly enrolled peer leaders and youth enrolled as peer educators for one or more years (repeat peer leaders). Quasi-experimental nonrandomized design with two intervention groups (newly enrolled and repeat peer leaders) and one comparison group. The sample consisted of 235 adolescents, 164 peer leaders, and 71 comparison youth, drawn from nine communities in Massachusetts. The intervention consisted of a short course and ongoing group work with an adult advisor to plan and implement HIV/AIDS outreach activities for youth. A confidential questionnaire administered at baseline and postintervention measured change in (a) HIV/AIDS knowledge, (b) planning and presenting skills, (c) self-efficacy, (d) perception of one's self as a change agent in the community, and (e) sexual risk-taking behaviors. Information was collected from both groups of peer leaders on specific activities resulting from the program and perceived benefits. Data were analyzed by both descriptive and multivariate statistics. Over a 9-month period newly enrolled peer leaders had significantly higher mean scores for HIV/AIDS knowledge and perception of one's self as a change agent in the community than comparison youth. On all baseline outcome measures except risk-taking behaviors, repeat peer leaders reported higher scores than newly enrolled peer leaders. Post-intervention, HIV/AIDS knowledge continued to increase significantly more among repeat peer leaders compared with those newly enrolled in the program. Repeat peer leaders also reported more benefits from peer leadership training. A peer education program was found to have benefits to adolescent peer leaders. Benefits gained from the program were sustained and enhanced over time as evidenced by repeat peer leaders included in the study.

  14. Development of HIV-1 rectal-specific microbicides and colonic tissue evaluation.

    Directory of Open Access Journals (Sweden)

    Charlene S Dezzutti

    Full Text Available The gastrointestinal tract is structurally and functionally different from the vagina. Thus, the paradigm of topical microbicide development and evaluation has evolved to include rectal microbicides (RMs. Our interest was to create unique RM formulations to safely and effectively deliver antiretroviral drugs to mucosal tissue. RMs were designed to include those that spread and coat all surfaces of the rectum and distal colon rapidly (liquid and those that create a deformable, erodible barrier and remain localized at the administration site (gel. Tenofovir (TFV (1% was formulated as an aqueous thermoreversible fluid and a carbopol-based aqueous hydrogel. Lipid-based liquid and gel formulations were prepared for UC781 (0.1% using isopropyl myristate and GTCC (Caprylic/Capric Triglycerides, respectively. Formulations were characterized for pH, viscosity, osmolality, and drug content. Pre-clinical testing incorporated ex vivo colonic tissue obtained through surgical resections and flexible sigmoidoscopy (flex sig. As this was the first time using tissue from both sources side-by-side, the ability to replicate HIV-1 was compared. Efficacy of the RM formulations was tested by applying the products with HIV-1 directly to polarized colonic tissue and following viral replication. Safety of the formulations was determined by MTT assay and histology. All products had a neutral pH and were isoosmolar. While HIV-1BaL and HIV-1JR-CSF alone and in the presence of semen had similar replication trends between surgically resected and flex sig tissues, the magnitude of viral replication was significantly better in flex sig tissues. Both TFV and UC781 formulations protected the colonic tissue, regardless of tissue source, from HIV-1 and retained tissue viability and architecture. Our in vitro and ex vivo results show successful formulation of unique RMs. Moreover, the results of flex sig and surgically resected tissues were comparable suggesting the incorporation

  15. Drugs + HIV, Learn the Link

    Medline Plus

    Full Text Available ... Drug Misuse Hepatitis (Viral) HIV/AIDS Mental Health Military Opioid Overdose Reversal with Naloxone (Narcan, Evzio) Pain ... is treatment, but no cure, at the present time. The virus (HIV) and the disease it causes ( ...

  16. Research Report: HIV/AIDS

    Science.gov (United States)

    ... Viral) HIV/AIDS Mental Health Military Opioid Overdose Reversal with Naloxone (Narcan, Evzio) Pain Prevention Recovery Substance ... or alcohol are more likely to have unsafe sex that might expose them to HIV and other ...

  17. Aligning vertical interventions to health systems: a case study of the HIV monitoring and evaluation system in South Africa

    Directory of Open Access Journals (Sweden)

    Kawonga Mary

    2012-01-01

    Full Text Available Abstract Background Like many low- and middle-income countries, South Africa established a dedicated HIV monitoring and evaluation (M&E system to track the national response to HIV/AIDS. Its implementation in the public health sector has however not been assessed. Since responsibility for health services management lies at the district (sub-national level, this study aimed to assess the extent to which the HIV M&E system is integrated with the overall health system M&E function at district level. This study describes implementation of the HIV M&E system, determines the extent to which it is integrated with the district health information system (DHIS, and evaluates factors influencing HIV M&E integration. Methods The study was conducted in one health district in South Africa. Data were collected through key informant interviews with programme and health facility managers and review of M&E records at health facilities providing HIV services. Data analysis assessed the extent to which processes for HIV data collection, collation, analysis and reporting were integrated with the DHIS. Results The HIV M&E system is top-down, over-sized, and captures a significant amount of energy and resources to primarily generate antiretroviral treatment (ART indicators. Processes for producing HIV prevention indicators are integrated with the DHIS. However processes for the production of HIV treatment indicators by-pass the DHIS and ART indicators are not disseminated to district health managers. Specific reporting requirements linked to ear-marked funding, politically-driven imperatives, and mistrust of DHIS capacity are key drivers of this silo approach. Conclusions Parallel systems that bypass the DHIS represent a missed opportunity to strengthen system-wide M&E capacity. Integrating HIV M&E (staff, systems and process into the health system M&E function would mobilise ear-marked HIV funding towards improving DHIS capacity to produce quality and timely HIV

  18. Evaluation of the dried blood spot filter paper technology and five testing strategies of HIV-1 and HIV-2 infections in West Africa

    NARCIS (Netherlands)

    Sarge-Njie, Ramu; Schim van der Loeff, Maarten; Ceesay, Saihou; Cubitt, David; Sabally, Saihou; Corrah, Tumani; Whittle, Hilton

    2006-01-01

    Simple robust approaches are needed to monitor the prevalence and incidence of HIV in Africa. The aim of this study was to evaluate the use of dried blood spot (DBS) as an alternative to serum or plasma for sentinel surveillance. Paired DBS and blood samples were obtained from 200 patients attending

  19. Managing dependencies in forest offset projects: toward a more complete evaluation of reversal risk

    Science.gov (United States)

    David M Cooley; Chrsitopher S Galik; Thomas P Holmes; Carolyn Kousky; Roger M Cooke

    2011-01-01

    Although forest carbon offsets can play an important role in the implementation of comprehensive climate policy, they also face an inherent risk of reversal. If such risks are positively correlated across projects, it can affect the integrity of larger project portfolios and potentially the entire offsets program. Here, we discuss three types of risks that could affect...

  20. Mild desalination demopilot : New normalization approach to effectively evaluate electrodialysis reversal technology

    NARCIS (Netherlands)

    Bisselink, R.; Schepper, W. de; Trampé, J.; Broek, W. van den; Pinel, P.; Krutko, A.; Groot, N.

    2016-01-01

    Key performance indicators for characterization of nanofiltration performance are well developed, similar key performance indicators for electrodialysis reversal are however underdeveloped. Under the E4Water project Dow Benelux BV and Evides Industriewater BV operate a pilot facility to compare both

  1. Evaluation of massive MIMO systems using time-reversal beamforming technique

    DEFF Research Database (Denmark)

    Mbeutcha, Marie; Fan, Wei; Hejselbæk, Johannes

    2016-01-01

    In this paper, we investigate the performance of a massive MIMO system using the time-reversal beamforming technique. The massive MIMO channels are simulated with ray-tracing at 3.5 GHz with a 200 MHz-bandwidth. We use a 64-element uniform cylindrical array as base station (BS) and we equip two...

  2. Evaluation of the toxicity and reversibility profile of the aqueous seed ...

    African Journals Online (AJOL)

    Hunteria umbellata (K. Schum.) Hallier f. (family: Apocynaceae) is reputed for the folkloric management of labour, pain and swellings, stomach ulcers, diabetes, obesity, and anaemia, with no scientific report of its toxicity and reversibility profile. The present study was, therefore, aimed at investigating the in vivo toxicity and ...

  3. Extensive evaluation of a seronegative participant in an HIV-1 vaccine trial as a result of false-positive PCR.

    Science.gov (United States)

    Schwartz, D H; Laeyendecker, O B; Arango-Jaramillo, S; Castillo, R C; Reynolds, M J

    1997-07-26

    In the USA, more than 2000 volunteers have received one or more experimental HIV-1 vaccines in phase I and II clinical trials, and there have been breakthrough HIV-1 infections among participants receiving vaccine and placebo. Serological diagnosis of new HIV-1 infections in vaccine-trial participants will become increasingly complicated as more viral components are used in vaccines. Recognition of this problem has led to a reliance on viral-genome measurement to distinguish vaccine-induced immunity from HIV-1 infection. Currently, quantitative RNA measurement is expensive, prone to contamination, and reliable only in laboratories certified by manufacturers or that have quality-control programmes. A high-risk vaccinee presented after an acute febrile episode and was tested for HIV-1 infection by reverse transcriptase (RT) PCR of viral RNA. Further extensive tests were required to clarify the HIV-1 infection and immune status of the vaccinee, including repeat RT-PCR, nested DNA PCR, western blot, lymphoproliferation assay, cytotoxic T-cell lysis, CD8-depleted co-culture, and HIV-1 challenge culture. Initial testing of plasma by RNA RT-PCR was reported as positive. This result was not confirmed by viral cultures, nested DNA PCR, western blot, or EIA. Additional RNA RT-PCR assays gave positive results from earlier occasions in the vaccine trial. Eventually, testing of all previously reactive samples by RNA RT-PCR was repeated in a quality-controlled laboratory, and confirmed the negative HIV-1 status of the individual. This case report exemplifies the difficulties with use of viral-genome measurement as a screening test to diagnose HIV-1 infection, particularly in individuals who have ever participated in HIV-1 vaccine trials. Monitoring of large numbers of phase III vaccinees by RNA RT-PCR will not be feasible. The design of efficacy trials for new vaccines should be in parallel with development of antibody-based diagnostic tests that are capable of differentiating

  4. Evaluation of supplemental testing with the Multispot HIV-1/HIV-2 Rapid Test and APTIMA HIV-1 RNA Qualitative Assay to resolve specimens with indeterminate or negative HIV-1 Western blots.

    Science.gov (United States)

    Linley, Laurie; Ethridge, Steven F; Oraka, Emeka; Owen, S Michele; Wesolowski, Laura G; Wroblewski, Kelly; Landgraf, Kenneth M; Parker, Monica M; Brinson, Myra; Branson, Bernard M

    2013-12-01

    The use of Western blot (WB) as a supplemental test after reactive sensitive initial assays can lead to inconclusive or misclassified HIV test results, delaying diagnosis. To determine the proportion of specimens reactive by immunoassay (IA) but indeterminate or negative by WB that could be resolved by alternative supplemental tests recommended under a new HIV diagnostic testing algorithm. Remnant HIV diagnostic specimens that were reactive on 3rd generation HIV-1/2 IA and either negative or indeterminate by HIV-1 WB from 11 health departments were tested with the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test (Multispot) and the Gen-Probe APTIMA HIV-1 RNA Qualitative Assay (APTIMA). According to the new testing algorithm, 512 (89.8%) specimens were HIV-negative, 55 (9.6%) were HIV-1 positive (including 19 [3.3%] that were acute HIV-1 and 9 [1.6%] that were positive for HIV-1 by Multispot but APTIMA-negative), 2 (0.4%) were HIV-2 positive, and 1 (0.2%) was HIV-positive, type undifferentiated. 47 (21.4%) of the 220 WB-indeterminate and 8 (2.3%) of the 350 WB-negative specimens were HIV-1 positive. Applying the new HIV diagnostic algorithm retrospectively to WB-negative and indeterminate specimens, the HIV infection status could be established for nearly all of the specimens. IA-reactive HIV-infected persons with WB-negative results had been previously misclassified as uninfected, and HIV diagnosis was delayed for those with WB-indeterminate specimens. These findings underscore the limitations of the WB to confirm HIV infection after reactive results from contemporary 3rd or 4th generation IAs that can detect HIV antibodies several weeks sooner than the WB. Published by Elsevier B.V.

  5. Nutrition evaluation in HIV seropositive patients using the ...

    African Journals Online (AJOL)

    Objectives: The study was designed to compare the nutritional status evaluation of PLWHA using the Malnutrition Universal Screening Tool (MUST) and Subjective Global Assessment (SGA) tool, and to also determine the cut-off values of MUST and SGA that corresponds to underweight with BMI (<18.5kg/m2) as the gold ...

  6. Doppler echocardiographic evaluation of HIV-positive patients in different stages of the disease

    Directory of Open Access Journals (Sweden)

    Werneck Guilherme Lobosco

    1999-01-01

    Full Text Available OBJETIVE: To evaluate by Doppler echocardiography (DE early abnormalities of ventricular function in HIV-positive patients, as well as other cardiac abnormalities that can be detected by this method, with special emphasis on mitral valve flow. METHODS: 84 HIV- positive patients, 59 with CD4 cell count >500/mm³ (Group A and 25 with CD4 cell count 500/mm³ had no abnormalities by DE. Patients with a more advanced infection (those with a CD4 cell count <500/mm³, had a significantly abnormal LV systolic function and a higher incidence of pericardial effusion and mitral regurgitation. Mitral valve inflow by Doppler did not indicate diastolic dysfunction.

  7. Evaluation of an External Quality Assessment Program for HIV Testing in Haiti, 2006–2011

    Science.gov (United States)

    Louis, Frantz Jean; Anselme, Renette; Ndongmo, Clement; Buteau, Josiane; Boncy, Jacques; Dahourou, Georges; Vertefeuille, John; Marston, Barbara; Balajee, S. Arunmozhi

    2016-01-01

    Objectives To evaluate an external quality assessment (EQA) program for human immunodeficiency virus (HIV) rapid diagnostics testing by the Haitian National Public Health Laboratory (French acronym: LNSP). Acceptable performance was defined as any proficiency testing (PT) score more than 80%. Methods The PT database was reviewed and analyzed to assess the testing performance of the participating laboratories and the impact of the program over time. A total of 242 laboratories participated in the EQA program from 2006 through 2011; participation increased from 70 laboratories in 2006 to 159 in 2011. Results In 2006, 49 (70%) laboratories had a PT score of 80% or above; by 2011, 145 (97.5%) laboratories were proficient (P quality of testing and allowed the LNSP to document improvements in the quality of HIV rapid testing over time. PMID:24225755

  8. Evaluation of adhesive properties of Candida albicans isolated from the oral cavity in HIV positive patients.

    Science.gov (United States)

    Macura, A B; Bort, A

    2001-01-01

    The aim of the work was to compare adhesive properties of Candida albicans strains isolated from the oral cavity in HIV+ vs. HIV- persons. The materials were Candida albicans strains and buccal epithelial cells isolated from both HIV+ and HIV- persons. We applied the in vitro adherence test, primarily described by Kimura and Pearsall and modified by Macura. The strongest adherence was found between both fungi and epithelial cells isolated from a HIV+ person. The adherence of C. albicans isolated from HIV+ patients was significantly stronger to epithelium collected from HIV+ than HIV- persons.

  9. Evaluation involvement of local HIV/AIDS non-governmental organisations in Benin

    Directory of Open Access Journals (Sweden)

    Maurice T. Agonnoude

    2016-06-01

    Full Text Available Background: For some years, non-governmental organisations (NGOs and civil society have become increasingly involved in the fight against the HIV/AIDS pandemic in Africa. But even though their role is well appreciated, their actions are perceived as ineffective because of a lack of monitoring and evaluation capacity.Objective: This paper aims to describe local HIV/AIDS NGOs’ involvement in evaluation and the characteristics of this involvement.Method: Descriptive analysis of data collected in questionnaires completed by 34 NGO executives (one per NGO.Results: Most NGOs do not have the minimal conditions required for positive and effective involvement in evaluations. In addition, funding agencies’ expectations for evaluations, total human resources as well as experience as NGO are contextual factors that explain most aspects of their involvement in evaluations.Conclusion: This study provides funding agencies, NGO leaders and all those interested in developing evaluation capacity in these NGOs to understand the extent of the task in this area. They must keep in mind that there is no solution for all, but that solutions must be adapted to the developmental level of each organisation.

  10. Administrative integration of vertical HIV monitoring and evaluation into health systems: a case study from South Africa.

    Science.gov (United States)

    Kawonga, Mary; Fonn, Sharon; Blaauw, Duane

    2013-01-24

    In light of an increasing global focus on health system strengthening and integration of vertical programmes within health systems, methods and tools are required to examine whether general health service managers exercise administrative authority over vertical programmes. To measure the extent to which general health service (horizontal) managers, exercise authority over the HIV programme's monitoring and evaluation (M&E) function, and to explore factors that may influence this exercise of authority. This cross-sectional survey involved interviews with 51 managers. We drew ideas from the concept of 'exercised decision-space' - traditionally used to measure local level managers' exercise of authority over health system functions following decentralisation. Our main outcome measure was the degree of exercised authority - classified as 'low', 'medium' or 'high' - over four M&E domains (HIV data collection, collation, analysis, and use). We applied ordinal logistic regression to assess whether actor type (horizontal or vertical) was predictive of a higher degree of exercised authority, independent of management capacity (training and experience), and M&E knowledge. Relative to vertical managers, horizontal managers had lower HIV M&E knowledge, were more likely to exercise a higher degree of authority over HIV data collation (OR 7.26; CI: 1.9, 27.4), and less likely to do so over HIV data use (OR 0.19; CI: 0.05, 0.84). A higher HIV M&E knowledge score was predictive of a higher exercised authority over HIV data use (OR 1.22; CI: 0.99, 1.49). There was no association between management capacity and degree of authority. This study demonstrates a HIV M&E model that is neither fully vertical nor integrated. The HIV M&E is characterised by horizontal managers producing HIV information while vertical managers use it. This may undermine policies to strengthen integrated health system planning and management under the leadership of horizontal managers.

  11. An evaluation of quality of life and its determinants among people living with HIV/AIDS from Southern Brazil

    Directory of Open Access Journals (Sweden)

    Susane Müller Klug Passos

    2015-04-01

    Full Text Available This cross-sectional study evaluated the quality of life and its associated factors among people living with HIV/AIDS at a regional reference center for the treatment of HIV/AIDS in southern Brazil. WHOQOL-HIV Bref, ASSIST 2.0, HAD Scale, and a questionnaire were used to assess 625 participants on quality of life, clinical and sociodemographic characteristics, drug use, depression and anxiety. Multivariate analysis was performed through linear regression. The lowest results for quality of life were associated with being female, age (< 47 years, low education levels, low socioeconomic class, unemployment, not having a stable relationship, signs of anxiety and depression, abuse or addiction of psychoactive substances, lack of perceived social support, never taking antiretroviral medication, lipodystrophy, comorbidities, HIV related hospitalizations and a CD4+ cell count less than 350. Psychosocial factors should be included in the physical and clinical evaluation given their strong association with quality of life domains.

  12. Improved Healing of Large, Osseous, Segmental Defects by Reverse Dynamization: Evaluation in a Sheep Model

    Science.gov (United States)

    2015-10-01

    greatest possible force generated during the full ovine gait cycle [8]. This showed that within 500N axial loading, the fixator remains within the...was determined. Moreover, negligible inter-fragmentary movement differences of the fracture gap occurred during repeated load- cycling to mimic the...stiffness (left) and interfragmentary movement (right) Page 6 Dynamization Model Reverse-Dynamization Model Figure 4 Life cycle axial cyclic testing (3

  13. Evaluation of New Reverse Osmosis Membranes for the Separation of Toxic Compounds from Wastewater

    Science.gov (United States)

    1976-06-01

    showed that sodium perborate , EDTA and BIZ flushes all restored the flux to 80-85% of the initial values. Each additive qave approximately equal results...separation of organic compounds using a simple test with sodium chloride solution. In addition, exploratory work was conducted on the potential application of...Reverse Osmosis 74 20 Performance of NS-1O0 Membranes when Tested with 500 ppm Sodium Chloride Solution at 600 psig and 250C 82 21 Removal of Isomers of

  14. Computational fluid dynamics evaluation of flow reversal treatment of giant basilar tip aneurysm.

    Science.gov (United States)

    Alnæs, Martin Sandve; Mardal, Kent-Andre; Bakke, Søren; Sorteberg, Angelika

    2015-10-01

    Therapeutic parent artery flow reversal is a treatment option for giant, partially thrombosed basilar tip aneurysms. The effectiveness of this treatment has been variable and not yet studied by applying computational fluid dynamics. Computed tomography images and blood flow velocities acquired with transcranial Doppler ultrasonography were obtained prior to and after bilateral endovascular vertebral artery occlusion for a giant basilar tip aneurysm. Patient-specific geometries and velocity waveforms were used in computational fluid dynamics simulations in order to determine the velocity and wall shear stress changes induced by treatment. Therapeutic parent artery flow reversal lead to a dramatic increase in aneurysm inflow and wall shear stress (30 to 170 Pa) resulting in an increase in intra-aneurysmal circulation. The enlargement of the circulated area within the aneurysm led to a re-normalization of the wall shear stress and the aneurysm remained stable for more than 8 years thereafter. Therapeutic parent artery flow reversal can lead to unintended, potentially harmful changes in aneurysm inflow which can be quantified and possibly predicted by applying computational fluid dynamics. © The Author(s) 2015.

  15. Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks.

    Science.gov (United States)

    Otani, Marcia M; Salles, Nanci A; Barreto, Angela M E; Barreto, Claudia C; Chamone, Dalton F; Sabino, Ester C

    2003-01-01

    In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV) concomitantly using two different enzyme immunoassay (EIA) tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of São Paulo (Fundação Pró-Sangue/Hemocentro de São Paulo), which is a major blood center in the city of São Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB) test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. Out of the 698 191 blood units that were donated, 2 718 of them (0.4%) had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58%) did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great majority of HIV-positive donors have already seroconverted and

  16. Factors associated with the use of irreversible contraception and continuous use of reversible contraception in a cohort of HIV-positive women

    NARCIS (Netherlands)

    Kancheva Landolt, Nadia; Ramautarsing, Reshmie Ashmanie; Phanuphak, Nittaya; Teeratakulpisarn, Nipat; Pinyakorn, Suteeraporn; Rodbamrung, Piyanee; Chaithongwongwatthana, Surasith; Ananworanich, Jintanat

    2013-01-01

    Effective contraception can be lifesaving by reducing maternal mortality linked to childbirth and unsafe abortion and by reducing vertical and horizontal transmission of HIV, in the case of an HIV-positive woman. This study is a secondary analysis of a prospective cohort study. We assessed factors

  17. Evaluation of a needle social marketing strategy to control HIV among injecting drug users in China.

    Science.gov (United States)

    Wu, Zunyou; Luo, Wei; Sullivan, Sheena G; Rou, Keming; Lin, Peng; Liu, Wei; Ming, Zhongqiang

    2007-12-01

    To evaluate the effectiveness of a needle social marketing strategy to reduce needle sharing and hepatitis C Virus (HCV)/HIV transmission among injecting drug users (IDU) in China. Two-armed, prospective, community-randomized prevention trial. Four counties/townships in Guangxi and Guangdong provinces; one randomized to intervention the other to control in each province. Injecting drug users: 823 (443 intervention, 382 control) at baseline and 852 (415 intervention, 407 control) at the second cross-sectional survey 12 months later. A needle social marketing programme, including promotion of safe injection norms and increased access to clean needles over a 12 month period. Cross sectional surveys at baseline and follow-up compared changes in drug using behaviours and HIV and HCV rates in the intervention and control communities. Needle sharing behaviours were similar in the two groups at baseline (68.4 vs. 67.8%), and dropped significantly to 35.3% in the intervention community and remained relatively stable in the control community (62.3%; P marketing can reduce risky injecting behaviour and HIV/HCV transmission among injecting drug users in China and should be expanded.

  18. Evaluation of a school-based HIV prevention intervention among Yemeni adolescents

    Directory of Open Access Journals (Sweden)

    Crutzen Rik

    2011-05-01

    Full Text Available Abstract Background This article describes an evaluation of a school-based peer education intervention for HIV prevention among students in twenty seven high schools in Aden, Yemen. The intervention was developed after a survey among the same population in 2005, which revealed a high level of stigma towards people living with HIV (PLWH and a low level of HIV knowledge. Methods In a quasi-experimental design students who received the peer education intervention (78.6% were compared with students who did not receive the intervention (21.4%. No systematic procedure was applied in selecting students for the intervention condition. Data were collected using a self-administered questionnaire from a sample of 2510 students from all 27 high-schools in Aden governorate. To increase internal validity, students were also compared with a cohort control sample surveyed in 2005, which was a random sample of 2274 students from the same schools. Results Sixty eight percent of students targeted by peer education had good knowledge scores, compared with 43.3% of students not targeted by peer education (χ2 = (df = 1 = 111.15, p Conclusion The school-based peer education intervention has succeeded in improving levels of knowledge on modes of transmission and prevention, and in decreasing levels of stigma and discrimination in a culturally conservative setting.

  19. Synthesis and evaluation of antiviral activities of novel sonochemical silver nanorods against HIV and HSV viruses

    Directory of Open Access Journals (Sweden)

    Mazyar Etemadzade

    2016-11-01

    Full Text Available Objective: To evaluate the effect of novel sonochemical silver nanorods on HIV and herpes simplex virus type 1 (HSV-1 viruses in human cervical cancer HeLa cells. Methods: The formation of silver nanorods conjugated with sodium 2-mercaptoethane sulfonate (Ag-MES was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and thermal gravimetric analysis. The antiviral activity of this Ag-MES was examined against HIV and HSV-1 virus replication. Results: The characterizations of Ag-MES and physiochemical structure were determined by scanning electron microscopy, Fourier transform infrared spectroscopy and thermal gravimetric analysis. Approximately entire viral replication was inhibited by Ag-MES at 10 µmol/mL concentration. About 90% of HSV virions failed to replicate in the present of this concentration of nanorods. However, HIV showed more sensitivity to Ag-MES than HSV-1. Conclusions: According to the obtained data, the synthesized sonochemical silver nanorod in this study is a promising candidate for further drug discovery investigation.

  20. Economic evaluation of universal prenatal HIV screening compared with current 'at risk' policy in a very low prevalence country.

    Science.gov (United States)

    Chowers, Michal; Shavit, Oren

    2017-03-01

    Our objective was to economically evaluate universal HIV prenatal screening in Israel, a very low prevalence country (0.1%), compared with the current policy of testing only women belonging to high-risk (HR) groups. A cost-effectiveness analytical model was constructed. Life expectancies, direct medical costs and utility weights of an HIV-positive newborn and a healthy newborn were derived from the literature. Screening was assessed using fourth-generation combo tests. Structural uncertainties were discussed with leading Israeli HIV experts. Univariate and multivariate sensitivity analyses were conducted to account for uncertainty of the model's parameters. Under the current policy, about 2700 women are tested annually identifying 27 HIV-positive women. With the universal screening, 171 000 women would be tested yearly identifying 37 as HIV positive. The analysis included the increased life expectancy of vertically infected children based on current standards of care. Over the lifetime expectancy, universal screening is projected to grant 15 additional quality-adjusted life years and save $177 521 when compared with the current HR only policy. Universal prenatal HIV screening is projected to be cost saving in Israel, despite a very low HIV prevalence in the general population. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  1. Reducing HIV risk among transgender women in Thailand: a quasi-experimental evaluation of the sisters program.

    Directory of Open Access Journals (Sweden)

    Duangta Pawa

    Full Text Available Transgender women are particularly at risk of HIV infection, but little evidence exists on effective HIV prevention strategies with this population. We evaluated whether Sisters, a peer-led program for transgender women, could reduce HIV risks in Pattaya, Thailand. The study used time-location sampling to recruit 308 transgender women in Pattaya into a behavioral survey in 2011. Coarsened exact matching was used to create statistically equivalent groups of program participants and non-participants, based on factors influencing likelihood of program participation. Using multivariable logistic regression, we estimated effects of any program participation and participation by delivery channel on: condom use at last sex; consistent condom and condom/water-based lubricant use in the past 3 months with commercial, casual, and regular partners; and receipt of HIV testing in the past 6 months. Program coverage reached 75% of the population. In a matched sub-sample (n = 238, participation in outreach was associated with consistent condom/water-based lubricant use with commercial partners (AOR 3.22, 95% CI 1.64-6.31. Attendance at the Sisters drop-in center was associated with receiving an HIV test (AOR 2.58, 95% CI 1.47-4.52. Dedicated transgender-friendly programs are effective at reducing HIV risks and require expansion to better serve this key population and improve HIV prevention strategies.

  2. Evaluation of sequence variability in HIV-1 gp41 C-peptide helix-grafted proteins.

    Science.gov (United States)

    Tennyson, Rachel L; Walker, Susanne N; Ikeda, Terumasa; Harris, Reuben S; McNaughton, Brian R

    2017-08-01

    Many therapeutically-relevant protein-protein interactions (PPIs) have been reported that feature a helix and helix-binding cleft at the interface. Given this, different approaches to disrupting such PPIs have been developed. While short peptides (<15 amino acids) typically do not fold into a stable helix, researchers have reported chemical approaches to constraining helix structure. However, these approaches rely on laborious, and often expensive, chemical synthesis and purification. Our premise is that protein-based solutions that stabilize a therapeutically-relevant helix offer a number of advantages. In contrast to chemically constrained helical peptides, or minimal/miniature proteins, which must be synthesized (at great expense and labor), a protein can be expressed in a cellular system (like all current protein therapeutics). If selected properly, the protein scaffold can stabilize the therapeutically-relevant helix. We recently reported a protein engineering strategy, which we call "helix-grafted display", and applied it to the challenge of suppressing HIV entry. We have reported helix-grafted display proteins that inhibit formation of an intramolecular PPI involving HIV gp41 C-peptide helix, and HIV gp41 N-peptide trimer, which contain C-peptide helix-binding clefts. Here, we used yeast display to screen a library of grafted C-peptide helices for N-peptide trimer recognition. Using 'hits' from yeast display library screening, we evaluated the effect helix mutations have on structure, expression, stability, function (target recognition), and suppression of HIV entry. Copyright © 2017. Published by Elsevier Ltd.

  3. Evaluation of a new generation synthetic peptide combination assay for detection of antibodies to HIV-1, HIV-2, HTLV-I, and HTLV-II simultaneously.

    Science.gov (United States)

    Zhang, X; Constantine, N T; Bansal, J; Callahan, J D; Marsiglia, V C

    1992-09-01

    A new generation combination test (Detect-Plus, IAF BioChem, Montreal, Canada) based on synthetic peptides for HIV-1, HIV-2, HTLV-I, and HTLV-II was compared with three routine commercial screening assays and confirmatory assays to determine its sensitivity and specificity and to evaluate it as a substitute screening method. Samples from 356 sexually transmitted disease (STD) patients were tested by the four screening tests. All initially reactive samples were retested in duplicate by the corresponding EIA and repeatedly reactive samples were confirmed by Western blots for HIV-1, HIV-2, and HTLV-I/II. The confirmed positives detected by each screening assay were HIV-1 (23/356, 6.46%), HIV-2 (11/356, 3.09%), and HTLV-I/II (5/356, 1.4%). The new generation Detect-Plus test produced only two results (2/356, 0.56%) that were presumed to be false-positives in comparison to the screening tests, but the OD/CO values were just slightly high (1.5 and 1.9). There were no false-negative results, indicating that the sensitivity of the new combination test was excellent (100%). Compared with routine retroviral EIA assays, the test is easy to perform--the total time requirement is only 2 hr and there is no need for incubation equipment. The OD/CO values were very high when samples were positive, making even visual interpretation possible. We conclude that this new combination assay is an excellent screening method for detection of antibodies to the human retroviruses, and may be particularly useful for screening blood for transfusion and in epidemiological investigations.

  4. Tighter binding of HIV reverse transcriptase to RNA-DNA vs. DNA-DNA results mostly from interactions in the polymerase domain and requires just a small stretch of RNA-DNA*

    OpenAIRE

    Bohlayer, William P.; DeStefano, Jeffrey J.

    2006-01-01

    Binding of HIV reverse transcriptase (RT) to unique substrates that positioned RNA-DNA or DNA-DNA near the polymerase or RNase H domains was measured. The substrates consisted of a 50 nucleotide template and DNA primers ranging from 23–43 nucleotides. Five different types of template strands were used: homogeneous (1) RNA or (2) DNA, (3) first 20 5′ nucleotides DNA and last 30 RNA, (4) first 20 RNA and last 30 DNA, (5) 15 nucleotides DNA followed by 5 RNA then 30 DNA. The different length pri...

  5. Evaluation of a new fourth-generation microwell enzyme-linked immunosorbent assay for detection of HIV-1 subtype B and E antibodies.

    Science.gov (United States)

    Chanbancherd, P; Limpairojn, N; de Souza, M S; Jugsudee, A; Julananto, P; Tienamporn, P; Leucha, W; Tasaniyananda, C; Brown, A E

    2001-03-01

    The recent fourth-generation enzyme-immunoassays have been used to increase the sensitivity for detecting HIV-1 antibodies and reduce the window period of HIV infection. The HIV antigens utilized in those assays were prepared from HIV-1 clade B which is different from HIV-1 subtypes circulating in Thailand. We evaluated 323 HIV-1 seropositives either B or E subtype to determine whether they were detected with the new combined anti-HIV and the p24 Ag assay. Under evaluation we found that this enzyme immunoassay manufactured by Organon Teknika showed the high sensitivity and specificity with a greater delta (delta) value with B than E subtypes samples (+15.29 vs +5.73).

  6. Process evaluation of school-based peer education for HIV prevention among Yemeni adolescents.

    Science.gov (United States)

    Al-Iryani, Buthaina; Basaleem, Huda; Al-Sakkaf, Khaled; Kok, Gerjo; van den Borne, Bart

    2013-03-01

    In 2005, a survey was conducted among all the 27 high schools of Aden, which revealed low levels of knowledge on major prevention measures, and a high level of stigma and discrimination towards people living with HIV (PLWH). The results served as a baseline for implementing a school-based peer education intervention for HIV prevention in the 27 schools of Aden. In 2008, and after 3 years of implementation, a quasi-experimental evaluation was conducted, which revealed that the peer education intervention has succeeded in improving HIV knowledge and skills; and in decreasing stigmatization of PLWH. This process evaluation aims to give a deeper understanding of the quasi-experimental evaluation which was conducted in the 27 high schools of Aden, and to highlight the factors that facilitated or inhibited school peer education in such a conservative Muslim setting. Qualitative methodologies were pursued, where 12 focus group discussions and 12 in-depth interviews were conducted with peer educators, targeted students, school principals, social workers, and parents of peer educators. Results revealed that school-peer education was well received. There was an apparent positive effect on the life skills of peer educators, but the intervention had a lesser effect on targeted students. Key enabling factors have been the high quality of training for peer educators, supportive school principals, and acceptance of the intervention by parents. These findings are important for improving the life skills and peer education intervention at the school level, and in better planning and implementation of life skills and peer programmes at a national scale.

  7. Evaluating potential artefacts of photo-reversal on behavioral studies with nocturnal invasive sea lamprey (Petromyzon marinus)

    Science.gov (United States)

    Barnett, Matthew; Imre, Istvan; Wagner, Michael C.; Di Rocco, Richard T.; Johnson, Nicholas; Brown, Grant E.

    2016-01-01

    Sea lampreys (Petromyzon marinus L., 1758) are nocturnal, so experiments evaluating their behaviour to chemosensory cues have typically been conducted at night. However, given the brief timeframe each year that adult P. marinus are available for experimentation, we investigated whether P. marinus exposed to a 12 h shifted diurnal cycle (reversed photoperiod) could be tested in a darkened arena during the day and show the same response to chemosensory cues as natural photoperiod P. marinus that were tested during the night. Ten replicates of 10 P. marinus, from each photoperiod, were exposed to deionized water (negative control), 2-phenylethylamine hydrochloride (PEA HCl, putative predator cue), or P. marinus whole-body extract (conspecific alarm cue). All P. marinus demonstrated a significant avoidance response to both cues. No significant differences were found in avoidance to PEA HCl between photoperiods. Avoidance of P. marinus whole-body extract was significantly stronger in natural compared with reversed photoperiod P. marinus. The use of reversed photoperiod subjects is suitable for examining the presence or absence of avoidance in response to novel chemosensory alarm cues, or the change in the magnitude of antipredator response. Studies investigating the natural magnitude of antipredator response should use natural photoperiod experimental subjects.

  8. Evaluation of four simple/rapid assays and two fourth-generation ELISAs for the identification of HIV infection on a serum panel representing the HIV-1 group M genetic diversity in Cameroon.

    Science.gov (United States)

    Aghokeng, Avelin Fobang; Ewane, Léonard; Awazi, Bih; Nanfack, Aubin; Delaporte, Eric; Peeters, Martine; Zekeng, Léopold

    2004-12-15

    The performance of 4 rapid and simple assays: Camstix-HIV 1+2 (Camdiagnostix, Yaounde, Cameroon); Determine HIV 1+2+0 (Abbott Laboratories, Tokyo, Japan); Genie II HIV-1/HIV-2 (Bio-Rad, Marnes la Coquette, France); ImmunoComb II HIV 1 & 2 BiSpot (Orgenics, Yavne, Israel); and 2 fourth-generation ELISAs: Enzygnost HIV Integral (Dade Behring, Marburg, Germany) and Genscreen plus HIV Ag-Ab (Bio-Rad, Marnes la Coquette, France) currently used in Cameroon to detect HIV infections were evaluated on a local serum panel. A total of 503 samples were collected, using the Camstix-HIV 1+2 assay. Overall, 280 samples were confirmed HIV positive, 181 were negative, and 42 were indeterminate. All positive samples belonged to group M: CRF02_AG (73.5%), A1 (7.1%), A2 (1.2%), G (4.7%), F2 (5.1%), D (1.6%), CRF11 (1.6%), CRF06 (1.2%), and CRF01_AE (1.6%). Sensitivity, specificity, test efficiency, and positive and negative predictive values were calculated both including and excluding indeterminate samples. Except for Genie II and ImmunoComb II (98.9 and 99.3%, respectively), sensitivities were 100% for the remaining 4 tests. Specificities, efficiencies, and positive predictive values of all assays were negatively affected by the addition of HIV-indeterminate samples in the calculations. These data show the importance of prior test evaluations on local serum panels and in field conditions before a national policy for HIV screening is decided on and stress also the need to use tests and algorithms that can reduce the high number of HIV-indeterminate results in Africa.

  9. Surface modification of seawater desalination reverse osmosis membranes: Characterization studies & performance evaluation

    KAUST Repository

    Matin, Asif

    2014-06-01

    In this work we report surface modification of commercial reverse osmosis membranes by depositing ultrathin copolymer coatings, which could potentially enhance the biofouling resistance of RO membranes. Hydrophilic monomer hydroxyethyl methacrylate (HEMA) and a hydrophobic monomer, perfluorodecyl acrylate (PFDA) were copolymerized directly on the active layer of commercial aromatic polyamide reverse osmosis (RO) membranes using an initiated Chemical Vapor Deposition (iCVD) technique. Attenuated total reflective Fourier transform infrared spectra (ATR-FTIR) verified the successful modification of the membrane surfaces as a new FTIR adsorption band around 1730cm-1 corresponding to carbonyl groups in the copolymer film appeared after the deposition. X-ray Photoelectron spectroscopy (XPS) analysis also confirmed the presence of the copolymer film on the membrane surface by showing strong fluorine peaks emanating from the fluorinated alkyl side chains of the PFA molecules. Contact angle measurements with deionized water showed the modified membrane surfaces to be initially very hydrophobic but quickly assumed a hydrophilic character within few minutes. Atomic Force Microscopy (AFM) revealed that the deposited films were smooth and conformal as the surface topology of the underlying membrane surface remained virtually unchanged after the deposition. FESEM images of the top surface also showed that the typical ridge-and-valley structure associated with polyamide remained intact after the deposition. Short-term permeation tests using DI water and 2000ppm NaCl water showed that the deposited copolymer coatings had negligible effect on permeate water flux and salt rejection. © 2013 Elsevier B.V.

  10. Evaluation of the use of reverse osmosis to eliminate natural radionuclides from water samples.

    Science.gov (United States)

    Nieto, Antonio; Palomo, Marta; Ruana, Josep; Peñalver, Alejandra; Aguilar, Carme; Borrull, Francesc

    2013-12-01

    The objective of drinking water treatment plants (DWTP) is to supply the population with tap water that is in optimal condition and in compliance with water quality regulations. In the DWTP of L'Ampolla (Tarragona, Spain), slightly high values of gross alpha activity and the amount of salts in the raw water have been observed. Conventional treatment has reduced these levels only minimally. This study tested a tertiary treatment based on reverse osmosis is tested in an industrial pilot plant (240 m3/day) The efficiency of this pilot plant to reduce the gross alpha and beta activities and the activity of some individual radioisotopes (U(238), U(234), U(235) and Ra(226)) was tested. Results showed that the elimination of alpha emitters was greater than 90%, whereas the elimination of beta emitters was about 35%. Overall, the data provided evidence that the pilot plant is effective for removing different radionuclides that can be present in the incoming water treated. Therefore, tertiary treatment based on reverse osmosis has a positive effect in water quality.

  11. Synthesis, in vitro evaluation, and docking studies of novel chromone derivatives as HIV-1 protease inhibitor

    Science.gov (United States)

    Ungwitayatorn, Jiraporn; Wiwat, Chanpen; Samee, Weerasak; Nunthanavanit, Patcharawee; Phosrithong, Narumol

    2011-08-01

    Novel chromone derivatives with a benzopyran-4-one scaffold have been prepared by the one-pot cyclization reaction. The in vitro inhibitory activity of these new compounds towards HIV-1 protease have been evaluated using stop time HPLC method as the preliminary screening. The most potent compound, 7,8-dihydroxy-2-(3'-trifluoromethyl phenyl)-3-(3″-trifluoromethylbenzoyl)chromone ( 32), showed IC 50 = 0.34 μM. The molecular docking study supported results from experimental activity testing and also provided structure-activity relationship of this series.

  12. Changes in liver steatosis evaluated by transient elastography with the controlled attenuation parameter in HIV-infected patients.

    Science.gov (United States)

    Macías, J; Real, L M; Rivero-Juárez, A; Merchante, N; Camacho, A; Neukam, K; Rivero, A; Mancebo, M; Pineda, J A

    2016-11-01

    There are scant data on the progression of hepatic steatosis (HS) in HIV infection. We therefore evaluated changes in HS over time in HIV-infected patients using the controlled attenuation parameter (CAP). A prospective cohort of 326 HIV-infected patients was included in this study. All patients underwent a CAP measurement. Changes in steatosis were evaluated by calculating the median (Q1-Q3) difference between baseline and 12-month CAP values. The median (Q1-Q3) CAP was 221 (196-252) dB/m at baseline and 224 (198-257) dB/m at the 12-month visit (P = 0.617). Significant steatosis, that is, CAP ≥ 238 dB/m, was observed in 76 individuals (37%) at baseline and in 80 (39%) at the 12-month visit (P = 0.683). The following variables were associated with ΔCAP: plasma HIV RNA [CAP values over a period of 12 months in HIV-infected patients were strongly associated with elevations in BMI. Other metabolic factors and antiretroviral drugs were not predictors of CAP changes independent of BMI. © 2016 British HIV Association.

  13. Prevalence of self-reported comorbidities in HIV positive and HIV negative men who have sex with men over 55 years-The Australian Positive & Peers Longevity Evaluation Study (APPLES.

    Directory of Open Access Journals (Sweden)

    Kathy Petoumenos

    Full Text Available In Australia, almost half of HIV-positive people are now aged over 50 and are predominately gay and bisexual men (GBM. Compared to the general HIV-negative population, GBM engage more in behaviours that may increase the risk of age-related comorbidities, including smoking, high alcohol consumption and recreational drug use. The objective of APPLES was to compare comorbidities and risk factors in HIV-positive older GBM with an appropriate control group of HIV-negative GBM. We undertook a prospectively recruited cross-sectional sample of HIV-positive and HIV-negative GBM ≥ 55 years. Detailed data collection included clinic data, a health and lifestyle survey, and blood sample collection. We report key demographic, laboratory markers and self-reported comorbidities by HIV status. For selected comorbidities we also adjust HIV status a priori for age, smoking and body mass index. Over 16 months 228 HIV-positive and 218 HIV-negative men were recruited. Median age was 63 years (IQR: 59-67. Although more HIV-positive men reported having ever smoked, smoking status was not statistically different between HIV positive and HIV negative men (p = 0.081. Greater alcohol use was reported by HIV-negative men (p = 0.002, and recreational drug use reported more often by HIV-positive men (p<0.001. After adjustment, HIV-positive men had significantly increased odds of diabetes (adjusted Odds ratio (aOR: 1.97, p = 0.038, thrombosis (aOR: 3.08, p = 0.007, neuropathy (aOR: 34.6, P<0.001, and non-significantly increased odds for heart-disease (aOR: 1.71, p = 0.077. In conclusion, HIV-positive GBM have significantly increased odds for key self-reported comorbidities. This study underscores the importance of an appropriate HIV-negative control group for more accurate evaluation of the risk and attribution of age-related comorbidities in HIV-positive people.

  14. [The Global Fund to fight HIV/AIDS, TB and Malaria 5-y: evaluation policy issues].

    Science.gov (United States)

    Kerouedan, D

    2010-05-01

    The Global Fund to fight HIV/AIDS, Tuberculosis and Malaria (GFATM) was founded in 2002 in the context of increased political and financial commitments towards health and development, in the aftermath of the Millennium Declaration, and on track to implement the Millennium Development Goals (MDGs). As of today, the institution has mobilized over 16 billion US dollars through its partnership, and spent over 8 billion dollars through 620 contracts in 140 countries for these three diseases. Principles at inception were to accelerate and expand HIV, TB, and Malaria prevention and awareness, care, and treatment related activities, in the poorest and the most affected countries worldwide, with a special emphasis on Africa, being the continent with the highest disease burden, especially with respect to HIV/AIDS and its dreadful social and economic consequences. In 2006, a Technical and Evaluation Reference Group was set up. This group responding to the GFATM Board in relation to the 5-year evaluation, defined the Terms of reference for the 5-year evaluation. Macro International, a firm based in Washington DC, was given the contract to conduct three studies over the period 2006-2009, looking at: (i) GFATM organizational effectiveness, (ii) partnerships at international and global levels, as well as systems effects, (iii) collective impact of the GFATM, the World Bank and (PEPFAR) funds on HIV, TB, and Malaria control. Twenty-five countries participated all together in the evaluation, out of which 18 in study area 3. Total budget for the evaluation amounted almost 17 million US dollars. This paper outlines: (i) the results of study areas 2 and 3 as well as the 5-year Evaluation Synthesis report, contents, and (ii) comments on the results and potential policy implications of the GFATM 5-year evaluation findings, as well as first responses prepared by the GF Secretariat shared at the GFATM Board meeting held in Ethiopia in November 2009. The evaluators raised the weaknesses of

  15. Developing a Motion Comic for HIV/STD Prevention for Young People Ages 15-24, Part 2: Evaluation of a Pilot Intervention.

    Science.gov (United States)

    Willis, Leigh A; Kachur, Rachel; Castellanos, Ted J; Nichols, Kristen; Mendoza, Maria C B; Gaul, Zaneta J; Spikes, Pilgrim; Gamayo, Ashley C; Durham, Marcus D; LaPlace, Lisa; Straw, Julie; Staatz, Colleen; Buge, Hadiza; Hogben, Matthew; Robinson, Susan; Brooks, John; Sutton, Madeline Y

    2018-03-01

    In the United States, young people (ages 15-24 years) are disproportionately affected by human immunodeficiency virus (HIV) and other sexually transmitted diseases (STDs), due at least in part to inadequate or incorrect HIV/STD-related knowledge, attitudes, beliefs, and behavioral intentions (KABI). Comic book narratives are a proven method of HIV/STD prevention communication to strengthen KABI for HIV/STD prevention. Motion comics, a new type of comic media, are an engaging and low-cost means of narrative storytelling. The objective of this study was to quantitatively evaluate the effectiveness of a pilot six-episode HIV/STD-focused motion comic series to improve HIV/STD-related KABI among young people. We assessed change in HIV/STD knowledge, HIV stigma, condom attitudes, HIV/STD testing attitudes, and behavioral intentions among 138 participants in 15 focus groups immediately before and after viewing the motion comic series. We used paired t-tests and indicators of overall improvement to assess differences between surveys. We found a significant decrease in HIV stigma (p comic intervention improved HIV/STD-related KABI of young adult viewers by reducing HIV stigma and increasing behavioral intentions to engage in safer sex. Our results demonstrate the promise of this novel intervention and support its use to deliver health messages to young people.

  16. Evaluation of screening kits for the detection of anti-human immunodeficiency virus type 1 and 2 (HIV-1/2) antibodies.

    Science.gov (United States)

    Chin, L T; Yang, B S; Chen, J W; Yang, C M; Chou, C C; Li, L; Hung, C M; Tsai, S J; Lin, K S

    1995-08-01

    HIV-1/HIV-2 3rd generation (Abbott), Wellcozyme HIV 1 + 2 (Murex), Enzygnost Anti-HIV 1/-HIV 2 (Behring), and Genelavia Mixt (Sanofi Diagnostics Pasteur) are currently registered by authorities as enzyme immunoassays (EIAs) for detecting HIV-1/2 infection. The present study dissects these reagents by means of the major antigenic components, assay principles and their actual performance. The performances have been evaluated by their test results in international panels of seroconversion, mixed titer performance and HIV-1/2 combination, respectively. Those EIA tests were further used to examine 26 potentially false-reacting samples, serial diluted sera prepared from two confirmed positive specimens and 720 specimens obtained from random blood donors in the Taipei Blood Center, Chinese Blood Services Foundation (CBSF). The results showed that, although standard sera of the mixed titer, performance and HIV-1/2 combination rows could not distinguish significantly among various EIAs, the seroconverting samples clearly showed their differences. The differences, as calculated by using 3 of 4 seroconverting sera, was a backward window period ranging from 19 to 23 days as compared to the detection of HIV-1 antigens. Together, these studies strongly suggest that assays which are capable of detecting HIV-specific IgM and IgG antibodies have a shorter seroconversion window. Furthermore, the HIV-2 antigen seems to be crucial for successful detection of anti-HIV-2. Finally, testing anti-HIV-1/2 in the routine screenings is expected not to increase the exclusion rate of blood units currently acquired from the examination of anti-HIV-1. Consequently, with both HIV-1/2 specificities and the ability of early detection, IgM/IgG-captured EIAs may represent a better screening method than assays based solely on the detection of HIV-specific IgG.

  17. Conventional HPLC Method Used for Simultaneous Determination of the Seven HIV Protease Inhibitors and Nonnucleoside Reverse Transcription Inhibitor Efavirenz in Human Plasma

    National Research Council Canada - National Science Library

    Masaaki TAKAHASHIa; b; Masao YOSHIDAa; Tsuyoshi OKIa; Naoya OKUMURAa; Tatsuo SUZUKIa; Tsuguhiro KANEDAb

    2005-01-01

    We developed a simple HPLC method for the simultaneous quantitative determination of seven HIV protease inhibitors:amprenavir (APV), atazanavir (ATV), indinavir (IDV), lopinavir (LPV), nelfinavir (NFV), ritonavir (RTV), saquinavir (SQV...

  18. Conventional HPLC Method Used for Simultaneous Determination of the Seven HIV Protease Inhibitors and Nonnucleoside Reverse Transcription Inhibitor Efavirenz in Human Plasma

    National Research Council Canada - National Science Library

    Takahashi, Masaaki; Yoshida, Masao; Oki, Tsuyoshi; Okumura, Naoya; Suzuki, Tatsuo; Kaneda, Tsuguhiro

    2005-01-01

    We developed a simple HPLC method for the simultaneous quantitative determination of seven HIV protease inhibitors: amprenavir (APV), atazanavir (ATV), indinavir (IDV), lopinavir (LPV), nelfinavir (NFV), ritonavir (RTV), saquinavir (SQV...

  19. Fused heterocycles bearing bridgehead nitrogen as potent HIV-1 NNRTIs. Part 4: design, synthesis and biological evaluation of novel imidazo[1,2-a]pyrazines.

    Science.gov (United States)

    Huang, Boshi; Liang, Xin; Li, Cuicui; Chen, Wenmin; Liu, Tao; Li, Xiao; Sun, Yueyue; Fu, Lu; Liu, Huiqing; De Clercq, Erik; Pannecouque, Christophe; Zhan, Peng; Liu, Xinyong

    2015-03-26

    Through a structure-guided core-refining approach, a series of novel imidazo[1,2-a]pyrazine derivatives were designed, synthesized and evaluated as HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs). Biological results of antiviral assay in MT-4 cell cultures showed that 12 target compounds displayed moderate activities against wild-type (wt) HIV-1 strain (IIIB) with EC50 values ranging from 0.26 μM to 19 μM. Among them, 4a and 5a were found to be the two most active analogues possessing EC50 values of 0.26 μM and 0.32 μM respectively, comparable to delavirdine (DLV, EC50 = 0.54 μM) and nevirapine (NVP, EC50 = 0.31 μM) in a cell-based assay. Additionally, 9 compounds showed RT inhibitory activity superior to that of NVP. Moreover, some predicted drug-like properties of representative compounds 4a and 5a, as well as the structure-activity relationship (SAR) analysis were discussed in detail. The binding mode of compound 4a was investigated by molecular simulation studies. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  20. Full Viral Suppression, Low-Level Viremia, and Quantifiable Plasma HIV-RNA at the End of Pregnancy in HIV-Infected Women on Antiretroviral Treatment.

    Science.gov (United States)

    Baroncelli, Silvia; Pirillo, Maria F; Tamburrini, Enrica; Guaraldi, Giovanni; Pinnetti, Carmela; Degli Antoni, Anna; Galluzzo, Clementina M; Stentarelli, Chiara; Amici, Roberta; Floridia, Marco

    2015-07-01

    There is limited information on full viral suppression and low-level HIV-RNA viremia in HIV-infected women at the end of pregnancy. We investigated HIV-RNA levels close to delivery in women on antiretroviral treatment in order to define rates of complete suppression, low-level viremia, and quantifiable HIV-RNA, exploring as potential determinants some clinical and viroimmunological variables. Plasma samples from a national study in Italy, collected between 2003 and 2012, were used. According to plasma HIV-RNA levels, three groups were defined: full suppression (target not detected), low-level viremia (target detected but HIV-RNA (≥37 copies/ml). Multivariable logistic regression was used to define determinants of full viral suppression and of quantifiable HIV-RNA. Among 107 women evaluated at a median gestational age of 35 weeks, 90 (84.1%) had HIV-RNA HIV-RNA was 109 copies/ml (IQR 46-251), with only one case showing resistance (mutation M184V; rate: 9.1%). In multivariable analyses, women with higher baseline HIV-RNA levels and with hepatitis C virus (HCV) coinfection were significantly more likely to have quantifiable HIV-RNA in late pregnancy. Full viral suppression was significantly more likely with nonnucleoside reverse transcriptase inhibitor (NNRTI)-based regimens and significantly less likely with higher HIV-RNA in early pregnancy. No cases of HIV transmission occurred. In conclusion, HIV-infected pregnant women showed a high rate of viral suppression and a low resistance rate before delivery. In most cases no target HIV-RNA was detected in plasma, suggesting a low risk of subsequent virological rebound and development of resistance. Women with high levels of HIV-RNA in early pregnancy and those who have concomitant HCV infection should be considered at higher risk of having quantifiable HIV-RNA at the end of pregnancy.

  1. Evaluation of GBV-C / HVG viremia in HIV-infected women

    Directory of Open Access Journals (Sweden)

    Synara Araújo Silva

    2012-02-01

    Full Text Available The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

  2. Evaluation of GBV-C / HVG viremia in HIV-infected women.

    Science.gov (United States)

    Silva, Synara Araújo; Rodrigues, Célia Lima; Campos, Aléia Faustino; Levi, José Eduardo

    2012-01-01

    The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

  3. An Outdated Notion of Antibody Specificity is One of the Major Detrimental Assumptions of the Structure-Based Reverse Vaccinology Paradigm, Which Prevented It from Helping to Develop an Effective HIV-1 Vaccine.

    Science.gov (United States)

    Van Regenmortel, Marc H V

    2014-01-01

    The importance of paradigms for guiding scientific research is explained with reference to the seminal work of Karl Popper and Thomas Kuhn. A prevalent paradigm, followed for more than a decade in HIV-1 vaccine research, which gave rise to the strategy known as structure-based reverse vaccinology is described in detail. Several reasons why this paradigm did not allow the development of an effective HIV-1 vaccine are analyzed. A major reason is the belief shared by many vaccinologists that antibodies possess a narrow specificity for a single epitope and are not polyspecific for a diverse group of potential epitopes. When this belief is abandoned, it becomes obvious that the one particular epitope structure observed during the crystallographic analysis of a neutralizing antibody-antigen complex does not necessarily reveal, which immunogenic structure should be used to elicit the same type of neutralizing antibody. In the physical sciences, scientific explanations are usually presented as logical deductions derived from a relevant law of nature together with certain initial conditions. In immunology, causal explanations in terms of a single cause acting according to a law of nature are not possible because numerous factors always play a role in bringing about an effect. The implications of this state of affairs for the rational design of HIV vaccines are outlined. An alternative approach to obtain useful scientific understanding consists in intervening empirically in the immune system and it is suggested that manipulating the system experimentally is needed to learn to control it and achieve protective immunity by vaccination.

  4. Hepatitis B virus sequencing and liver fibrosis evaluation in HIV/HBV co-infected Nigerians.

    Science.gov (United States)

    Grant, Jennifer; Agbaji, Oche; Kramvis, Anna; Yousif, Mukhlid; Auwal, Mu'azu; Penugonda, Sudhir; Ugoagwu, Placid; Murphy, Robert; Hawkins, Claudia

    2017-06-01

    Molecular characteristics of hepatitis B virus (HBV), such as genotype and genomic mutations, may contribute to liver-related morbidity and mortality. The association of these characteristics with liver fibrosis severity in sub-Saharan Africa is uncertain. We aimed to characterise molecular HBV features in human immunodeficiency virus (HIV)/HBV co-infected Nigerians and evaluate associations between these characteristics and liver fibrosis severity before and after antiretroviral therapy (ART) initiation. HIV/HBV co-infected Nigerians underwent liver fibrosis estimation by transient elastography (TE) prior to and 36 months after ART initiation. Basal core promoter/precore (BCP/PC) and preS1/preS2/S regions of HBV were sequenced from baseline plasma samples. We evaluated associations between HBV mutations and liver fibrosis severity by univariate and multivariable regression. At baseline, 94 patients underwent TE with median liver stiffness of 6.4 (IQR 4.7-8.7) kPa. Patients were predominantly infected with HBV genotype E (45/46) and HBe-antigen negative (75/94, 79.8%). We identified BCP A1762T/G1764A in 15/35 (43%), PC G1896A in 20/35 (57%), 'a' determinant mutations in 12/45 (26.7%) and preS2 deletions in 6/16 (37.5%). PreS2 mutations were associated with advanced fibrosis in multivariable analysis. At follow-up, median liver stiffness was 5.2 (IQR 4.1-6.6) kPa. No HBV molecular characteristics were associated with lack of fibrosis regression, although HIV virologic control, body mass index (BMI) and baseline CD4+ T-cell count were associated with a decline in fibrosis stage. Frequent BCP/PC and preS1/preS2/S mutations were found in ART-naïve HIV/HBV co-infected Nigerians. Median liver stiffness declined after initiation of ART, regardless of pre-ART HBV mutational pattern or virologic characteristics. © 2017 John Wiley & Sons Ltd.

  5. Iterative Evaluation in a Mobile Counseling and Testing Program to Reach People of Color at Risk for HIV--New Strategies Improve Program Acceptability, Effectiveness, and Evaluation Capabilities

    Science.gov (United States)

    Spielberg, Freya; Kurth, Ann; Reidy, William; McKnight, Teka; Dikobe, Wame; Wilson, Charles

    2011-01-01

    This article highlights findings from an evaluation that explored the impact of mobile versus clinic-based testing, rapid versus central-lab based testing, incentives for testing, and the use of a computer counseling program to guide counseling and automate evaluation in a mobile program reaching people of color at risk for HIV. The program's…

  6. HIV-related stigma and universal testing and treatment for HIV prevention and care: design of an implementation science evaluation nested in the HPTN 071 (PopART) cluster-randomized trial in Zambia and South Africa.

    Science.gov (United States)

    Hargreaves, James R; Stangl, Anne; Bond, Virginia; Hoddinott, Graeme; Krishnaratne, Shari; Mathema, Hlengani; Moyo, Maureen; Viljoen, Lario; Brady, Laura; Sievwright, Kirsty; Horn, Lyn; Sabapathy, Kalpana; Ayles, Helen; Beyers, Nulda; Bock, Peter; Fidler, Sarah; Griffith, Sam; Seeley, Janet; Hayes, Richard

    2016-12-01

    Stigma and discrimination related to HIV and key populations at high risk of HIV have the potential to impede the implementation of effective HIV prevention and treatment programmes at scale. Studies measuring the impact of stigma on these programmes are rare. We are conducting an implementation science study of HIV-related stigma in communities and health settings within a large, pragmatic cluster-randomized trial of a universal testing and treatment intervention for HIV prevention in Zambia and South Africa and will assess how stigma affects, and is affected by, implementation of this intervention. A mixed-method evaluation will be nested within HIV prevention trials network (HPTN) 071/PopART (Clinical Trials registration number NCT01900977), a three-arm trial comparing universal door-to-door delivery of HIV testing and referral to prevention and treatment services, accompanied by either an immediate offer of anti-retroviral treatment to people living with HIV regardless of clinical status, or an offer of treatment in-line with national guidelines, with a standard-of-care control arm. The primary outcome of HPTN 071/PopART is HIV incidence measured among a cohort of 52 500 individuals in 21 study clusters. Our evaluation will include integrated quantitative and qualitative data collection and analysis in all trial sites. We will collect quantitative data on indicators of HIV-related stigma over 3 years from large probability samples of community members, health workers and people living with HIV. We will collect qualitative data, including in-depth interviews and observations from members of these same groups sampled purposively. In analysis, we will: (1) compare HIV-related stigma measures between study arms, (2) link data on stigma to measures of the success of implementation of the PopART intervention and (3) explore changes in the dominant drivers and manifestations of stigma in study communities and the health system. HIV-related stigma may impede the

  7. Performance Evaluation of Reverse Osmosis Technology for Selected Antibiotics Removal from Synthetic Pharmaceutical Wastewater

    Directory of Open Access Journals (Sweden)

    Mitra Gholami

    2012-12-01

    Full Text Available This study addresses the possibility for low pressure reverse osmosis membrane (RE 2521, CSM process to serve as an alternative to remove selected antibiotics (ampicillin and amoxicillin from synthetic wastewater by changing operating conditions such as pH = 3, 6.5 and 10; Pressure = 9, 11 and13 (bar; antibiotic concentration = 10, 255and 500(mg/L, and temperature = 20, 30 and 40°C. The experiment was designed based on Box-benken, which is a Response Surface methodology design (RSM, using Design Expert software. The concentration of antibiotics was measured by applying a UV-spectrophotometer (Cecil, at the wavelength of 254 nm. Results showed a range ofrejection percentage from 73.52% to 99.36% and 75.1% to 98.8%, for amoxicillin and ampicillin, respectively.Considering the solute rejections and the membrane porosity show that the prevailing rejection mechanism of the examined antibiotics by the membrane was the size exclusion effect. The permeate flux for both of the antibiotics was 12–18.73 L/m2.h. Although the permeate flux and antibiotic rejection are influenced by operating pressure, pH,and temperature individually, the interaction between operating parameters did not have noticeable effects. According to the results obtained in this study, the application of RO membrane is recommended for the selected antibiotics to be removed to a considerable degree (up to 95%.

  8. Performance evaluation of reverse osmosis technology for selected antibiotics removal from synthetic pharmaceutical wastewater

    Science.gov (United States)

    2012-01-01

    This study addresses the possibility for low pressure reverse osmosis membrane (RE 2521, CSM) process to serve as an alternative to remove selected antibiotics (ampicillin and amoxicillin) from synthetic wastewater by changing operating conditions such as pH = 3, 6.5 and 10; Pressure = 9, 11 and13 (bar); antibiotic concentration = 10, 255 and 500(mg/L), and temperature = 20, 30 and 40°C. The experiment was designed based on Box-benken, which is a Response Surface methodology design (RSM), using Design Expert software. The concentration of antibiotics was measured by applying a UV-spectrophotometer (Cecil), at the wavelength of 254 nm. Results showed a range of rejection percentage from 73.52% to 99.36% and 75.1% to 98.8%, for amoxicillin and ampicillin, respectively. Considering the solute rejections and the membrane porosity show that the prevailing rejection mechanism of the examined antibiotics by the membrane was the size exclusion effect. The permeate flux for both of the antibiotics was 12–18.73 L/m2.h. Although the permeate flux and antibiotic rejection are influenced by operating pressure, pH, and temperature individually, the interaction between operating parameters did not have noticeable effects. According to the results obtained in this study, the application of RO membrane is recommended for the selected antibiotics to be removed to a considerable degree (up to 95%). PMID:23369431

  9. Evaluation of potential for reuse of industrial wastewater using metal-immobilized catalysts and reverse osmosis.

    Science.gov (United States)

    Choi, Jeongyun; Chung, Jinwook

    2015-04-01

    This report describes a novel technology of reusing the wastewater discharged from the display manufacturing industry through an advanced oxidation process (AOP) with a metal-immobilized catalyst and reverse osmosis (RO) in the pilot scale. The reclaimed water generated from the etching and cleaning processes in display manufacturing facilities was low-strength organic wastewater and was required to be recycled to secure a water source. For the reuse of reclaimed water to ultrapure water (UPW), a combination of solid-phase AOP and RO was implemented. The removal efficiency of TOC by solid-phase AOP and RO was 92%. Specifically, the optimal acid, pH, and H2O2 concentrations in the solid-phase AOP were determined. With regard to water quality and operating costs, the combination of solid-phase AOP and RO was superior to activated carbon/RO and ultraviolet AOP/anion polisher/coal carbon. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Performance evaluation of reverse osmosis technology for selected antibiotics removal from synthetic pharmaceutical wastewater

    Directory of Open Access Journals (Sweden)

    Gholami Mitra

    2012-12-01

    Full Text Available Abstract This study addresses the possibility for low pressure reverse osmosis membrane (RE 2521, CSM process to serve as an alternative to remove selected antibiotics (ampicillin and amoxicillin from synthetic wastewater by changing operating conditions such as pH = 3, 6.5 and 10; Pressure = 9, 11 and13 (bar; antibiotic concentration = 10, 255 and 500(mg/L, and temperature = 20, 30 and 40°C. The experiment was designed based on Box-benken, which is a Response Surface methodology design (RSM, using Design Expert software. The concentration of antibiotics was measured by applying a UV-spectrophotometer (Cecil, at the wavelength of 254 nm. Results showed a range of rejection percentage from 73.52% to 99.36% and 75.1% to 98.8%, for amoxicillin and ampicillin, respectively. Considering the solute rejections and the membrane porosity show that the prevailing rejection mechanism of the examined antibiotics by the membrane was the size exclusion effect. The permeate flux for both of the antibiotics was 12–18.73 L/m2.h. Although the permeate flux and antibiotic rejection are influenced by operating pressure, pH, and temperature individually, the interaction between operating parameters did not have noticeable effects. According to the results obtained in this study, the application of RO membrane is recommended for the selected antibiotics to be removed to a considerable degree (up to 95%.

  11. Impact and economic evaluation of a novel HIV service delivery model in rural Malawi.

    Science.gov (United States)

    McBain, Ryan K; Petersen, Elizabeth; Tophof, Nora; Dunbar, Elizabeth L; Kalanga, Noel; Nazimera, Lawrence; Mganga, Andrew; Dullie, Luckson; Mukherjee, Joia; Wroe, Emily B

    2017-09-10

    We performed an impact and cost-effectiveness analysis of a novel HIV service delivery model in a high prevalence, remote district of Malawi with a population of 143 800 people. A population-based retrospective analysis of 1-year survival rates among newly enrolled HIV-positive patients at 682 health facilities throughout Malawi, comparing facilities implementing the service delivery model (n = 13) and those implementing care-as-usual (n = 669). Through district-level health surveillance data, we evaluated 1-year survival rates among HIV patients newly enrolled between July 2013 and June 2014 - representing 129 938 patients in care across 682 health facilities - using a multilevel modeling framework. The model, focused on social determinants of health, was implemented throughout Neno District at 13 facilities and compared with facilities in all other districts. Activity-based costing was used to annualize financial and economic costs from a societal perspective. Incremental cost-effectiveness ratios were expressed as quality-adjusted life-years gained. The national average 1-year survival rate for newly enrolled antiretroviral therapy clients was 78.9%: this rate was 87.9% in Neno District, compared with 78.8% across all other districts in Malawi (P cost of receiving care in Neno district (n = 6541 patients) was $317/patient/year, compared with an estimated $219/patient in other districts. This translated to $906 per quality-adjusted life-year gained. Neno District's comprehensive model of care, featuring a strong focus on the community, is $98 more expensive per capita per annum but demonstrates superior 1-year survival rates, despite its remote location. Moreover, it should be considered cost-effective by traditional international standards.

  12. Research Protocol - An Evaluation of False Positive HIV Results due to Testing Errors

    OpenAIRE

    Maparo, Tatenda; Mungofa, Stanley; Bara, Hilda T.; Chirisa, Florence

    2014-01-01

    An unacceptably high frequency of false positive HIV test results has been reported in various settings. Given the severity and implications of an HIV+ diagnosis, a false positive result is likely to be psychologically traumatic and may result in inappropriate and potentially harmful treatment. The current HIV testing algorithm being used in Zimbabwe does not include repeat testing for HIV positive results, and it is not currently known whether testing errors are leading to false positive dia...

  13. A flexible model of HIV-1 latency permitting evaluation of many primary CD4 T-cell reservoirs.

    Science.gov (United States)

    Lassen, Kara G; Hebbeler, Andrew M; Bhattacharyya, Darshana; Lobritz, Michael A; Greene, Warner C

    2012-01-01

    Latently infected cells form the major obstacle to HIV eradication. Studies of HIV latency have been generally hindered by the lack of a robust and rapidly deployable cell model that involves primary human CD4 T lymphocytes. Latently infected cell lines have proven useful, but it is unclear how closely these proliferating cells recapitulate the conditions of viral latency in non-dividing CD4 T lymphocytes in vivo. Current primary lymphocyte models more closely reflect the in vivo state of HIV latency, but they are limited by protracted culture periods and often low cell yields. Additionally, these models are always established in a single latently infected cell type that may not reflect the heterogeneous nature of the latent reservoir. Here we describe a rapid, sensitive, and quantitative primary cell model of HIV-1 latency with replication competent proviruses and multiple reporters to enhance the flexibility of the system. In this model, post-integration HIV-1 latency can be established in all populations of CD4 T cells, and reactivation of latent provirus assessed within 7 days. The kinetics and magnitude of reactivation were evaluated after stimulation with various cytokines, small molecules, and T-cell receptor agonists. Reactivation of latent HIV proviruses was readily detected in the presence of strong activators of NF-κB. Latently infected transitional memory CD4 T cells proved more responsive to these T-cell activators than latently infected central memory cells. These findings reveal potentially important biological differences within the latently infected pool of memory CD4 T cells and describe a flexible primary CD4 T-cell system to evaluate novel antagonists of HIV latency.

  14. Evaluation of HIV and AIDS knowledge in rural Cameroon men with the use of a questionnaire

    NARCIS (Netherlands)

    H.P. Versteegh (Hendt); A. Bakia (Affuenti); H.M. Koopman (Hendrik); V. Kraaij (Vivian); F.G. Versteegh (Florens)

    2013-01-01

    textabstractIntroduction: HIV/AIDS, the most important health problem in Africa, is the leading cause of death on the continent. Ignorance on HIV/AIDS status will hamper treatment and prevention. To investigate the level of HIV/AIDS knowledge among men in a rural area, we performed a questionnaire

  15. Design and pre-clinical evaluation of a universal HIV-1 vaccine.

    Directory of Open Access Journals (Sweden)

    Sven Létourneau

    2007-10-01

    Full Text Available One of the big roadblocks in development of HIV-1/AIDS vaccines is the enormous diversity of HIV-1, which could limit the value of any HIV-1 vaccine candidate currently under test.To address the HIV-1 variation, we designed a novel T cell immunogen, designated HIV(CONSV, by assembling the 14 most conserved regions of the HIV-1 proteome into one chimaeric protein. Each segment is a consensus sequence from one of the four major HIV-1 clades A, B, C and D, which alternate to ensure equal clade coverage. The gene coding for the HIV(CONSV protein was inserted into the three most studied vaccine vectors, plasmid DNA, human adenovirus serotype 5 and modified vaccine virus Ankara (MVA, and induced HIV-1-specific T cell responses in mice. We also demonstrated that these conserved regions prime CD8(+ and CD4(+ T cell to highly conserved epitopes in humans and that these epitopes, although usually subdominant, generate memory T cells in patients during natural HIV-1 infection.Therefore, this vaccine approach provides an attractive and testable alternative for overcoming the HIV-1 variability, while focusing T cell responses on regions of the virus that are less likely to mutate and escape. Furthermore, this approach has merit in the simplicity of design and delivery, requiring only a single immunogen to provide extensive coverage of global HIV-1 population diversity.

  16. Evaluation of prevention of mother-to-child hiv transmission program ...

    African Journals Online (AJOL)

    The results showed that among 3774 antenatal attendees, 2528 (67%) accepted pre-test counselling and 2390 (63%) HIV testing. Majority (95%) of those who had (2528) pre-test counselling accepted HIV testing, post test counselling and test results. The prevalence of HIV infection was 41% (980) (95% CI, 39%-43%).

  17. The Development of a Test System for the Evaluation of Reverse Osmosis Water Purification Membranes

    Science.gov (United States)

    1984-06-01

    The absence of chlorine residual was confirmed by regular determinations with NN-diethyl-p-phenylene- diamine (DPD) according to Standard Methods for the...changed initially. The performance evaluation sample supplied with the test column contains azobenzene , octadecane and ma•.athion in 2,2,4...trimethylpentane. Evaluation of performance is based on the ratios of azobenzene and malathion peak I..,. areas to the octadecane peak area and the ratios of these

  18. Drugs + HIV, Learn the Link

    Medline Plus

    Full Text Available ... Consequences of Drug Misuse Hepatitis (Viral) HIV/AIDS Mental Health Military Opioid Overdose Reversal with Naloxone (Narcan, Evzio) ... HIV or transmitting it to someone else. Biological effects of drugs. Drug ... overall health, thereby altering susceptibility to HIV and progression of ...

  19. An In-Depth Comparison of Latency-Reversing Agent Combinations in Various In Vitro and Ex Vivo HIV-1 Latency Models Identified Bryostatin-1+JQ1 and Ingenol-B+JQ1 to Potently Reactivate Viral Gene Expression.

    Directory of Open Access Journals (Sweden)

    Gilles Darcis

    2015-07-01

    Full Text Available The persistence of latently infected cells in patients under combinatory antiretroviral therapy (cART is a major hurdle to HIV-1 eradication. Strategies to purge these reservoirs are needed and activation of viral gene expression in latently infected cells is one promising strategy. Bromodomain and Extraterminal (BET bromodomain inhibitors (BETi are compounds able to reactivate latent proviruses in a positive transcription elongation factor b (P-TEFb-dependent manner. In this study, we tested the reactivation potential of protein kinase C (PKC agonists (prostratin, bryostatin-1 and ingenol-B, which are known to activate NF-κB signaling pathway as well as P-TEFb, used alone or in combination with P-TEFb-releasing agents (HMBA and BETi (JQ1, I-BET, I-BET151. Using in vitro HIV-1 post-integration latency model cell lines of T-lymphoid and myeloid lineages, we demonstrated that PKC agonists and P-TEFb-releasing agents alone acted as potent latency-reversing agents (LRAs and that their combinations led to synergistic activation of HIV-1 expression at the viral mRNA and protein levels. Mechanistically, combined treatments led to higher activations of P-TEFb and NF-κB than the corresponding individual drug treatments. Importantly, we observed in ex vivo cultures of CD8+-depleted PBMCs from 35 cART-treated HIV-1+ aviremic patients that the percentage of reactivated cultures following combinatory bryostatin-1+JQ1 treatment was identical to the percentage observed with anti-CD3+anti-CD28 antibodies positive control stimulation. Remarkably, in ex vivo cultures of resting CD4+ T cells isolated from 15 HIV-1+ cART-treated aviremic patients, the combinations bryostatin-1+JQ1 and ingenol-B+JQ1 released infectious viruses to levels similar to that obtained with the positive control stimulation. The potent effects of these two combination treatments were already detected 24 hours post-stimulation. These results constitute the first demonstration of LRA

  20. Performance evaluation of a reverse-gradient artificial recharge system in basalt aquifers of Maharashtra, India

    Science.gov (United States)

    Bhusari, Vijay; Katpatal, Y. B.; Kundal, Pradeep

    2017-05-01

    Drinking water scarcity in rural parts of central India in basaltic terrain is common. Most of the rural population depends on groundwater sources located in the fractured and weathered zone of the basaltic aquifers. Long-term indiscriminate withdrawal has caused an alarming rate of depletion of groundwater levels in both pre- and post-monsoon periods. The aquifer is not replenished through precipitation under natural conditions. To overcome this situation, an innovative artificial recharge system, called the reverse-gradient recharge system (RGRS), was implemented in seven villages of Wardha district of Maharashtra. The study described here presents a comparative analysis of recharge systems constructed in the year 2012 downstream of dug-well locations in these seven villages. The post-project comparative analysis reveals that the area of influence (AOI) of the groundwater recharge system, within which increases in groundwater levels and yield are observed, is directly related to the specific yield, thickness of the weathered and fractured zone, porosity, and transmissivity of the aquifer, showing high correlation coefficients of 0.92, 0.88, 0.85 and 0.83, respectively. The study indicates that the RGRS is most effective in vesicular weathered and fractured basalt, recording a maximum increase in well yield of 65-82 m3/day, while a minimum increase in yield of 15-30 m3/day was observed in weathered vesicular basalt. The comparative analysis thus identifies the controlling factors which facilitate groundwater recharge through the proposed RGRS. After implementation of these projects, the groundwater availability in these villages increased significantly, solving their drinking water problems.

  1. Reverse Engineering and Security Evaluation of Commercial Tags for RFID-Based IoT Applications

    Science.gov (United States)

    Fernández-Caramés, Tiago M.; Fraga-Lamas, Paula; Suárez-Albela, Manuel; Castedo, Luis

    2016-01-01

    The Internet of Things (IoT) is a distributed system of physical objects that requires the seamless integration of hardware (e.g., sensors, actuators, electronics) and network communications in order to collect and exchange data. IoT smart objects need to be somehow identified to determine the origin of the data and to automatically detect the elements around us. One of the best positioned technologies to perform identification is RFID (Radio Frequency Identification), which in the last years has gained a lot of popularity in applications like access control, payment cards or logistics. Despite its popularity, RFID security has not been properly handled in numerous applications. To foster security in such applications, this article includes three main contributions. First, in order to establish the basics, a detailed review of the most common flaws found in RFID-based IoT systems is provided, including the latest attacks described in the literature. Second, a novel methodology that eases the detection and mitigation of such flaws is presented. Third, the latest RFID security tools are analyzed and the methodology proposed is applied through one of them (Proxmark 3) to validate it. Thus, the methodology is tested in different scenarios where tags are commonly used for identification. In such systems it was possible to clone transponders, extract information, and even emulate both tags and readers. Therefore, it is shown that the methodology proposed is useful for auditing security and reverse engineering RFID communications in IoT applications. It must be noted that, although this paper is aimed at fostering RFID communications security in IoT applications, the methodology can be applied to any RFID communications protocol. PMID:28029119

  2. Reverse Engineering and Security Evaluation of Commercial Tags for RFID-Based IoT Applications

    Directory of Open Access Journals (Sweden)

    Tiago M. Fernández-Caramés

    2016-12-01

    Full Text Available The Internet of Things (IoT is a distributed system of physical objects that requires the seamless integration of hardware (e.g., sensors, actuators, electronics and network communications in order to collect and exchange data. IoT smart objects need to be somehow identified to determine the origin of the data and to automatically detect the elements around us. One of the best positioned technologies to perform identification is RFID (Radio Frequency Identification, which in the last years has gained a lot of popularity in applications like access control, payment cards or logistics. Despite its popularity, RFID security has not been properly handled in numerous applications. To foster security in such applications, this article includes three main contributions. First, in order to establish the basics, a detailed review of the most common flaws found in RFID-based IoT systems is provided, including the latest attacks described in the literature. Second, a novel methodology that eases the detection and mitigation of such flaws is presented. Third, the latest RFID security tools are analyzed and the methodology proposed is applied through one of them (Proxmark 3 to validate it. Thus, the methodology is tested in different scenarios where tags are commonly used for identification. In such systems it was possible to clone transponders, extract information, and even emulate both tags and readers. Therefore, it is shown that the methodology proposed is useful for auditing security and reverse engineering RFID communications in IoT applications. It must be noted that, although this paper is aimed at fostering RFID communications security in IoT applications, the methodology can be applied to any RFID communications protocol.

  3. Reverse Engineering and Security Evaluation of Commercial Tags for RFID-Based IoT Applications.

    Science.gov (United States)

    Fernández-Caramés, Tiago M; Fraga-Lamas, Paula; Suárez-Albela, Manuel; Castedo, Luis

    2016-12-24

    The Internet of Things (IoT) is a distributed system of physical objects that requires the seamless integration of hardware (e.g., sensors, actuators, electronics) and network communications in order to collect and exchange data. IoT smart objects need to be somehow identified to determine the origin of the data and to automatically detect the elements around us. One of the best positioned technologies to perform identification is RFID (Radio Frequency Identification), which in the last years has gained a lot of popularity in applications like access control, payment cards or logistics. Despite its popularity, RFID security has not been properly handled in numerous applications. To foster security in such applications, this article includes three main contributions. First, in order to establish the basics, a detailed review of the most common flaws found in RFID-based IoT systems is provided, including the latest attacks described in the literature. Second, a novel methodology that eases the detection and mitigation of such flaws is presented. Third, the latest RFID security tools are analyzed and the methodology proposed is applied through one of them (Proxmark 3) to validate it. Thus, the methodology is tested in different scenarios where tags are commonly used for identification. In such systems it was possible to clone transponders, extract information, and even emulate both tags and readers. Therefore, it is shown that the methodology proposed is useful for auditing security and reverse engineering RFID communications in IoT applications. It must be noted that, although this paper is aimed at fostering RFID communications security in IoT applications, the methodology can be applied to any RFID communications protocol.

  4. Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks

    Directory of Open Access Journals (Sweden)

    Otani Marcia M.

    2003-01-01

    Full Text Available OBJECTIVE: In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV concomitantly using two different enzyme immunoassay (EIA tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. METHODS: We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of São Paulo (Fundação Pró-Sangue/Hemocentro de São Paulo, which is a major blood center in the city of São Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. RESULTS: Out of the 698 191 blood units that were donated, 2 718 of them (0.4% had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58% did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. CONCLUSIONS: Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great

  5. Comparative evaluation of the Bio-Rad Geenius HIV-1/2 Confirmatory Assay and the Bio-Rad Multispot HIV-1/2 Rapid Test as an alternative differentiation assay for CLSI M53 algorithm-I.

    Science.gov (United States)

    Malloch, L; Kadivar, K; Putz, J; Levett, P N; Tang, J; Hatchette, T F; Kadkhoda, K; Ng, D; Ho, J; Kim, J

    2013-12-01

    The CLSI-M53-A, Criteria for Laboratory Testing and Diagnosis of Human Immunodeficiency Virus (HIV) Infection; Approved Guideline includes an algorithm in which samples that are reactive on a 4th generation EIA screen proceed to a supplemental assay that is able to confirm and differentiate between antibodies to HIV-1 and HIV-2. The recently CE-marked Bio-Rad Geenius HIV-1/2 Confirmatory Assay was evaluated as an alternative to the FDA-approved Bio-Rad Multispot HIV-1/HIV-2 Rapid Test which has been previously validated for use in this new algorithm. This study used reference samples submitted to the Canadian - NLHRS and samples from commercial sources. Data was tabulated in 2×2 tables for statistical analysis; sensitivity, specificity, predictive values, kappa and likelihood ratios. The overall performance of the Geenius and Multispot was very high; sensitivity (100%, 100%), specificity (96.3%, 99.1%), positive (45.3, 181) and negative (0, 0) likelihood ratios respectively, high kappa (0.96) and low bias index (0.0068). The ability to differentiate HIV-1 (99.2%, 100%) and HIV-2 (98.1%, 98.1%) Ab was also very high. The Bio-Rad Geenius HIV-1/2 Confirmatory Assay is a suitable alternative to the validated Multispot for use in the second stage of CLSI M53 algorithm-I. The Geenius has additional features including traceability and sample and cassette barcoding that improve the quality management/assurance of HIV testing. It is anticipated that the CLSI M53 guideline and assays such as the Geenius will reduce the number of indeterminate test results previously associated with the HIV-1 WB and improve the ability to differentiate HIV-2 infections. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  6. Total HIV/AIDS expenditures in Dehong Prefecture, Yunnan province in 2010: the first systematic evaluation of both health and non-health related HIV/AIDS expenditures in China.

    Directory of Open Access Journals (Sweden)

    Duo Shan

    Full Text Available We assessed HIV/AIDS expenditures in Dehong Prefecture, Yunnan Province, one of the highest prevalence regions in China, and describe funding sources and spending for different categories of HIV-related interventions and at-risk populations.2010 HIV/AIDS expenditures in Dehong Prefecture were evaluated based on UNAIDS' National AIDS Spending Assessment methodology.Nearly 93% of total expenditures for HIV/AIDS was contributed by public sources. Of total expenditures, 52.7% was allocated to treatment and care, 24.5% to program management and administration and 19.8% to prevention. Spending on treatment and care was primarily allocated to the treatment of opportunistic infections. Most (40.4% prevention spending was concentrated on most-at-risk populations, injection drug users (IDUs, sex workers, and men who have sex with men (MSM, with 5.5% allocated to voluntary counseling and testing. Prevention funding allocated for MSM, partners of people living with HIV and prisoners and other confined populations was low compared to the disproportionate burden of HIV/AIDS in these populations. Overall, people living with HIV accounted for 57.57% of total expenditures, while most-at-risk populations accounted for only 7.99%.Our study demonstrated the applicability of NASA for tracking and assessing HIV expenditure in the context of China, it proved to be a useful tool in understanding national HIV/AIDS response from financial aspect, and to assess the extent to which HIV expenditure matches epidemic patterns. Limited funding for primary prevention and prevention for MSM, prisoners and partners of people living with HIV, signal that resource allocation to these key areas must be strengthened. Comprehensive analyses of regional and national funding strategies are needed to inform more equitable, effective and cost-effective HIV/AIDS resource allocation.

  7. Reverse Logistics

    OpenAIRE

    Kulikova, Olga

    2016-01-01

    This thesis was focused on the analysis of the concept of reverse logistics and actual reverse processes which are implemented in mining industry and finding solutions for the optimization of reverse logistics in this sphere. The objective of this paper was the assessment of the development of reverse logistics in mining industry on the example of potash production. The theoretical part was based on reverse logistics and mining waste related literature and provided foundations for further...

  8. Evaluation of Project RISE, an HIV Prevention Intervention for Black Bisexual Men Using an Ecosystems Approach.

    Science.gov (United States)

    Lauby, Jennifer; Milnamow, Mary; Joseph, Heather A; Hitchcock, Shannon; Carson, Lee; Pan, Yi; Mendoza, Maria; Millett, Greg

    2017-09-04

    Black men who have sex with men and women (MSMW) are among the populations at highest risk for HIV infection. We describe the evaluation of Project RISE, a six-session individual-level intervention developed for black MSMW using an ecosystems approach. A randomized controlled trial was used to test the effect of the intervention on sexual risk outcomes. Eligibility criteria included having both male and female sex partners in the past 12 months. Complete data at 5-month follow-up were collected from 86.7% of the 165 participants. In analyses controlling for HIV status, age, and baseline risk, intervention participants were found to have significantly greater reductions in number of female partners (p < 0.05) and total male and female partners (p < 0.05) at follow-up, compared to the control group. Intervention participants also were significantly more likely to report a reduction in number of sex episodes without a condom with female partners (p < 0.05) and with all partners (p < 0.02) at follow-up, compared to the control group.

  9. Approach to Cerebrospinal Fluid (CSF) Biomarker Discovery and Evaluation in HIV Infection

    Energy Technology Data Exchange (ETDEWEB)

    Price, Richard W.; Peterson, Julia; Fuchs, Dietmar; Angel, Thomas E.; Zetterberg, Henrik; Hagberg, Lars; Spudich, Serena S.; Smith, Richard D.; Jacobs, Jon M.; Brown, Joseph N.; Gisslen, Magnus

    2013-12-13

    Central nervous system (CNS) infection is a nearly universal facet of systemic HIV infection that varies in character and neurological consequences. While clinical staging and neuropsychological test performance have been helpful in evaluating patients, cerebrospinal fluid (CSF) biomarkers present a valuable and objective approach to more accurate diagnosis, assessment of treatment effects and understanding of evolving pathobiology. We review some lessons from our recent experience with CSF biomarker studies. We have used two approaches to biomarker analysis: targeted, hypothesis-driven and non-targeted exploratory discovery methods. We illustrate the first with data from a cross-sectional study of defined subject groups across the spectrum of systemic and CNS disease progression and the second with a longitudinal study of the CSF proteome in subjects initiating antiretroviral treatment. Both approaches can be useful and, indeed, complementary. The first is helpful in assessing known or hypothesized biomarkers while the second can identify novel biomarkers and point to broad interactions in pathogenesis. Common to both is the need for well-defined samples and subjects that span a spectrum of biological activity and biomarker concentrations. Previouslydefined guide biomarkers of CNS infection, inflammation and neural injury are useful in categorizing samples for analysis and providing critical biological context for biomarker discovery studies. CSF biomarkers represent an underutilized but valuable approach to understanding the interactions of HIV and the CNS and to more objective diagnosis and assessment of disease activity. Both hypothesis-based and discovery methods can be useful in advancing the definition and use of these biomarkers.

  10. INTENTION TO USE CONDOM, CUSP MODELING, AND EVALUATION OF AN HIV PREVENTION INTERVNETION TRIAL

    Science.gov (United States)

    Chen, Xinguang; Stanton, Bonita; Chen, Ding-Geng; Li, Xiaoming

    2014-01-01

    Adolescents are at particularly high risk to acquire HIV infection; increasing the likelihood of condom use is an effective measure to reduce the risk of such infections. Challenges in assessing actual condom use behavior among early adolescents render the precursor measure, intention to use condoms, an appealing alternative. While analyzing data from a randomized controlled trial to evaluate a theory-based intervention program to promote condom use among early adolescents, we observed a modest effect with regard to condom use intention when the linear analytical approach was used. If intention, as a measure of the readiness to perform a behavior, also contains a nonlinear discrete component, it would be more appropriately modeled using a non-linear approach. In this study, data from a randomized controlled trial (N=1360) were analyzed using the cusp catastrophe method with HIV knowledge and condom skills as the asymmetry variables and condom use self-efficacy as the bifurcation variables. Findings from concurrent and longitudinal modeling analyses indicated a much better fit of the cusp model (R2 = 0.85 and 4+ times smaller AIC and BIC) than the linear (R2 intention but did not as a bifurcation variable. In conclusion, adolescent intentions to use a condom contain both a continuous process and a discrete process and can better be modeled with cusp methods. A much greater program effect is likely from the same prevention intervention if additional measures are taken to foster sudden changes in condom intention. PMID:23735493

  11. Evaluation of seroepidemiological toxoplasmosis in HIV/AIDS patients in the south of Brazil.

    Science.gov (United States)

    Xavier, Graciela Augusto; Cademartori, Beatris Gonzalez; Cunha Filho, Nilton Azevedo da; Farias, Nara Amélia da Rosa

    2013-01-01

    Toxoplasmosis is considered one of the opportunistic infections for individuals with the Acquired Immunodeficiency Syndrome (AIDS), and is also a major cause of morbidity and mortality. The aim of this study was to evaluate the prevalence of neurotoxoplasmosis, ocular toxoplasmosis and antibodies for Toxoplasma gondii in HIV-positive patients attending the SAE (Specialized Assistance Service for HIV/AIDS), as well as to associate their serological profile with epidemiological and clinical data. A total of 250 patients participated in the study from December, 2009 to November, 2010. Serological analysis was performed using the indirect immunofluorescent technique; epidemiological data were gathered by a questionnaire, and clinical history was based on the analysis of medical charts. Prevalence of seropositivity was 80%, with history of neurotoxoplasmosis in 4.8% and of ocular toxoplasmosis in 1.6% of the patients. The Highly Active Antiretroviral Treatment (HAART) was not used by 32% of the patients, 18.4% of the patients had CD4+ T- lymphocyte count less than 200 cells/mm³ and 96.8% of them were not aware of the modes of disease transmission. These findings led us to conclude that the study population is at high risk of clinical toxoplasmosis, because of both reactivation of infection in the seropositive patients who do not make a regular use of HAART, and primo-infection in seronegative patients worsened by an unawareness of the modes of infection reported in this study.

  12. Evaluating Patient Interest in an Adherence-Focused Smartphone App to Improve HIV Care

    Directory of Open Access Journals (Sweden)

    Joshua W Gaborcik

    2017-02-01

    Full Text Available Objective: Evaluate patient interest in a smartphone mobile application (app to assist in medication adherence. Methods: In January 2014, a 19-question, anonymous, paper survey was distributed to a convenience sample of patients in the reception area of a nonprofit HIV primary care clinic and pharmacy. Results: Of the 101 patients surveyed, 72.3% had a smartphone and 70.3% were interested in downloading and using an adherence app if one was available. If an app was customizable, patients desired appointment reminders (87%, notifications to schedule appointments (85%, refill notifications (83%, medication reminders (79%, and adherence tracked by pharmacy (59%. Conclusions: Results share insights on the potential use of technology to assist an HIV patient population with medication adherence. Conflict of Interest Dr. Jennifer Rodis is the creator and director of the Partner For Promotion (PFP program otherwise she has no additional conflicts of interest or financial interests that the authors or members of their immediate families have in any product or service discussed in the manuscript, including grants (pending or received, employment, gifts, stock holdings or options, honoraria, consultancies, expert testimony, patents and royalties. All other authors declare no conflicts of interest or financial interests that the authors or members of their immediate families have in any product or service discussed in the manuscript, including grants (pending or received, employment, gifts, stock holdings or options, honoraria, consultancies, expert testimony, patents and royalties   Type: Student Project

  13. Evaluating complications of local anesthesia administration and reversal with phentolamine mesylate in a portable pediatric dental clinic.

    Science.gov (United States)

    Boynes, Sean G; Riley, Amah E; Milbee, Sarah; Bastin, Meghan R; Price, Maylyn E; Ladson, Andrea

    2013-08-01

    This study sought to identify and quantify complications with local anesthetic administration and reversal on consecutive patients seen for comprehensive dental care in a school-based, portable dental clinic, and includes data on the patients seen by the participating portable dental providers. In 923 dental visits where local anesthetic was administered, a standardized form was used to gain further information and identify any complications; this was accompanied by a questionnaire for the student's teacher, in order to quantify the student's distraction and disruption ratings following the dental visit. After statistical analysis of the 923 consecutive cases, the overall complication rate was 5.3%. All of the complications were considered to be mild or moderate, and there were no severe event reports. The complications encountered most frequently (n = 49) were associated with self-inflicted soft tissue injury. The results of this study indicate that comprehensive care with local anesthesia delivered by a school-based portable dental clinic has a low risk of complications. Whereas safe administration of dental care is achievable with or without phentolamine mesylate as a local anesthetic reversal agent, its use was determined to improve safety outcomes. Three factors appeared to directly increase the incidence of complications: the administration of an inferior alveolar nerve block, attention deficit disorder, and obesity. Teacher evaluations demonstrated that children receiving care by a portable dental team were able to reorient back to classwork and were not disruptive to classmates.

  14. Efficient in vitro inhibition of HIV-1 gag reverse transcription by peptide nucleic acid (PNA) at minimal ratios of PNA/RNA

    DEFF Research Database (Denmark)

    Koppelhus, Uffe; Zachar, Vladimir; Nielsen, P.E.

    1997-01-01

    of reverse transcription was obtained at approximately 6-fold molar excess of the bis-PNA with respect to the gag RNA. At this molar ratio we found no effect on in vitro translation of gag RNA. A 15mer duplex-forming PNA was also found to inhibit reverse transcription at very low molar ratios of PNA/ gag RNA...... that would indicate PNA-mediated RNase H activation of the tested RTs. In conclusion, PNA appears to have a potential to become a specific and efficient inhibitor of reverse transcription in vivo , provided sufficient intracellular levels are achievable....

  15. Administrative integration of vertical HIV monitoring and evaluation into health systems: a case study from South Africa

    Directory of Open Access Journals (Sweden)

    Mary Kawonga

    2013-01-01

    Full Text Available Background: In light of an increasing global focus on health system strengthening and integration of vertical programmes within health systems, methods and tools are required to examine whether general health service managers exercise administrative authority over vertical programmes. Objective: To measure the extent to which general health service (horizontal managers, exercise authority over the HIV programme's monitoring and evaluation (M&E function, and to explore factors that may influence this exercise of authority. Methods: This cross-sectional survey involved interviews with 51 managers. We drew ideas from the concept of ‘exercised decision-space’ – traditionally used to measure local level managers’ exercise of authority over health system functions following decentralisation. Our main outcome measure was the degree of exercised authority – classified as ‘low’, ‘medium’ or ‘high’ – over four M&E domains (HIV data collection, collation, analysis, and use. We applied ordinal logistic regression to assess whether actor type (horizontal or vertical was predictive of a higher degree of exercised authority, independent of management capacity (training and experience, and M&E knowledge. Results: Relative to vertical managers, horizontal managers had lower HIV M&E knowledge, were more likely to exercise a higher degree of authority over HIV data collation (OR 7.26; CI: 1.9, 27.4, and less likely to do so over HIV data use (OR 0.19; CI: 0.05, 0.84. A higher HIV M&E knowledge score was predictive of a higher exercised authority over HIV data use (OR 1.22; CI: 0.99, 1.49. There was no association between management capacity and degree of authority. Conclusions: This study demonstrates a HIV M&E model that is neither fully vertical nor integrated. The HIV M&E is characterised by horizontal managers producing HIV information while vertical managers use it. This may undermine policies to strengthen integrated health system

  16. Evaluating an HIV and AIDS Community Training Partnership Program in five diamond mining communities in South Africa.

    Science.gov (United States)

    Rispel, L C; Peltzer, K; Nkomo, N; Molomo, B

    2010-11-01

    In 2006, De Beers Consolidated Diamond Mines in South Africa entered into a partnership with the Soul City Institute for Health and Development Communications to implement an HIV and AIDS Community Training Partnership Program (CTPP), initially in five diamond mining areas in three provinces of South Africa. The aim of CTPP was to improve HIV knowledge and to contribute to positive behavior changes in the targeted populations. This paper describes the evaluation of the CTPP, one year after implementation. The evaluation combined qualitative interviews with key informants and trainers and a post-intervention survey of 142 community members. The successes of the CTPP included capacity building of trainers through an innovative training approach and HIV and AIDS knowledge transfer to community trainers and targeted communities in remote mining towns. The Soul City edutainment brand is popular and emerged as a major reason for success. Challenges included insufficient attention paid to contextual factors, resource constraints and the lack of a monitoring and evaluation framework. Independent evaluations are useful to strengthen program implementation. In remote areas and resource constraint settings, partnerships between non-governmental organisations and corporations may be required for successful community HIV and AIDS initiatives. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  17. Ribonuclease H/DNA Polymerase HIV-1 Reverse Transcriptase Dual Inhibitor: Mechanistic Studies on the Allosteric Mode of Action of Isatin-Based Compound RMNC6.

    Directory of Open Access Journals (Sweden)

    Angela Corona

    Full Text Available The DNA polymerase and ribonuclease H (RNase H activities of human immunodeficiency virus type 1 (HIV-1 are needed for the replication of the viral genome and are validated drug targets. However, there are no approved drugs inhibiting RNase H and the efficiency of DNA polymerase inhibitors can be diminished by the presence of drug resistance mutations. In this context, drugs inhibiting both activities could represent a significant advance towards better anti-HIV therapies. We report on the mechanisms of allosteric inhibition of a newly synthesized isatin-based compound designated as RMNC6 that showed IC50 values of 1.4 and 9.8 μM on HIV-1 RT-associated RNase H and polymerase activities, respectively. Blind docking studies predict that RMNC6 could bind two different pockets in the RT: one in the DNA polymerase domain (partially overlapping the non-nucleoside RT inhibitor [NNRTI] binding pocket, and a second one close to the RNase H active site. Enzymatic studies showed that RMNC6 interferes with efavirenz (an approved NNRTI in its binding to the RT polymerase domain, although NNRTI resistance-associated mutations such as K103N, Y181C and Y188L had a minor impact on RT susceptibility to RMNC6. In addition, despite being naturally resistant to NNRTIs, the polymerase activity of HIV-1 group O RT was efficiently inhibited by RMNC6. The compound was also an inhibitor of the RNase H activity of wild-type HIV-1 group O RT, although we observed a 6.5-fold increase in the IC50 in comparison with the prototypic HIV-1 group M subtype B enzyme. Mutagenesis studies showed that RT RNase H domain residues Asn474 and Tyr501, and in a lesser extent Ala502 and Ala508, are critical for RMNC6 inhibition of the endonuclease activity of the RT, without affecting its DNA polymerization activity. Our results show that RMNC6 acts as a dual inhibitor with allosteric sites in the DNA polymerase and the RNase H domains of HIV-1 RT.

  18. Ribonuclease H/DNA Polymerase HIV-1 Reverse Transcriptase Dual Inhibitor: Mechanistic Studies on the Allosteric Mode of Action of Isatin-Based Compound RMNC6.

    Science.gov (United States)

    Corona, Angela; Meleddu, Rita; Esposito, Francesca; Distinto, Simona; Bianco, Giulia; Masaoka, Takashi; Maccioni, Elias; Menéndez-Arias, Luis; Alcaro, Stefano; Le Grice, Stuart F J; Tramontano, Enzo

    2016-01-01

    The DNA polymerase and ribonuclease H (RNase H) activities of human immunodeficiency virus type 1 (HIV-1) are needed for the replication of the viral genome and are validated drug targets. However, there are no approved drugs inhibiting RNase H and the efficiency of DNA polymerase inhibitors can be diminished by the presence of drug resistance mutations. In this context, drugs inhibiting both activities could represent a significant advance towards better anti-HIV therapies. We report on the mechanisms of allosteric inhibition of a newly synthesized isatin-based compound designated as RMNC6 that showed IC50 values of 1.4 and 9.8 μM on HIV-1 RT-associated RNase H and polymerase activities, respectively. Blind docking studies predict that RMNC6 could bind two different pockets in the RT: one in the DNA polymerase domain (partially overlapping the non-nucleoside RT inhibitor [NNRTI] binding pocket), and a second one close to the RNase H active site. Enzymatic studies showed that RMNC6 interferes with efavirenz (an approved NNRTI) in its binding to the RT polymerase domain, although NNRTI resistance-associated mutations such as K103N, Y181C and Y188L had a minor impact on RT susceptibility to RMNC6. In addition, despite being naturally resistant to NNRTIs, the polymerase activity of HIV-1 group O RT was efficiently inhibited by RMNC6. The compound was also an inhibitor of the RNase H activity of wild-type HIV-1 group O RT, although we observed a 6.5-fold increase in the IC50 in comparison with the prototypic HIV-1 group M subtype B enzyme. Mutagenesis studies showed that RT RNase H domain residues Asn474 and Tyr501, and in a lesser extent Ala502 and Ala508, are critical for RMNC6 inhibition of the endonuclease activity of the RT, without affecting its DNA polymerization activity. Our results show that RMNC6 acts as a dual inhibitor with allosteric sites in the DNA polymerase and the RNase H domains of HIV-1 RT.

  19. Design, Synthesis, and Biological Evaluation of Novel Cholesteryl Peptides with Anticancer and Multidrug Resistance-Reversing Activities.

    Science.gov (United States)

    Deng, Xin; Qiu, Qianqian; Wang, Xuekun; Huang, Wenlong; Qian, Hai

    2016-03-01

    Antimicrobial peptides have been suggested as promising chemotherapeutics for cancer therapy due to their efficient antitumor activity and lower toxicity to benign cells. In previous study, we find the peptide B1 presents specific cytotoxicity to cancer cells. As hydrophobicity plays a pivotal role in the anticancer activity of peptide, we introduce cholesterol-like moiety (3β-amino-5-cholestene) to the N-terminus of B1 expect to ameliorate the anticancer activity of B1. Biological evaluations revealed that target peptides show improved anticancer activity. The peptides can also penetrate into the cytoplasm and activating mitochondria-cytochrome c apoptosis pathway. Besides, the peptides acted on multidrug-resistant cells and had multidrug resistance-reversing activity. It is therefore suggested these peptides might be promising candidates for oncotherapy. © 2015 John Wiley & Sons A/S.

  20. [Total reverse shoulder replacement. Evaluation of the clinical results and complications in a series of 52 cases].

    Science.gov (United States)

    Cáceres-Sánchez, L; Mesa-Mateo, A; Barrionuevo-Sánchez, F J; García-Benítez, B; Expósito-Triano, S

    2015-01-01

    To evaluate the clinical results and analyse the complications of total reverse shoulder replacement performed in our centre over an 8 year period. A retrospective study was conducted on 50 patients (52 shoulders), with a mean age of 70.15 years (range 51 to 84 years) between December 2004 and December 2012, who received a total reverse shoulder replacement, all performed by the same surgeon. The results have been evaluated according to clinical data, radiography study, a satisfaction scale, and the Constant scale, with a minimum follow-up of 16 months. Five of the cases (9.62%) had been intervened due to fractures of the proximal end of the humerus, 6 cases (11.53%) as surgical consequence of a prosthesis revision, 10 cases (19.23%) due to fracture sequelae, and 30 cases (59.62%) were patients with arthropathy due to a massive fracture of the rotator cuff. After a mean follow up of 35.78 months (range, 16-82), satisfactory clinical results were obtained in 80% of cases, with a mean preoperative Constant of 27.7 points, and reaching 67.1 points 12 months after the operation. On the visual analogue scale, 8.25 points were obtained before the surgery, which decreased to 2.25 points 12 months later. The complications rate was 15.38%, which were due to an intra-operative fracture (1.92%), deep infection (3.84%), instability (3.84%), and early mechanical loosening (3.84%). Scapular notching was observed in the radiographic study in 9 (17.3%) cases. After the results obtained, it could be said that total reverse shoulder replacement achieved encouraging results in the short term for the treatment of glenohumeral arthrosis and massive tears of the rotary cuff. On analysing our series, it can be seen that the complications rate is much higher when it is used to treat fracture sequelae in which there is a loss of proximal humerus bone stock. Copyright © 2014 SECOT. Published by Elsevier Espana. All rights reserved.

  1. Prognostic evaluation of DNA index in HIV-HPV co-infected women cervical samples attending in reference centers for HIV-AIDS in Recife.

    Directory of Open Access Journals (Sweden)

    Albert Eduardo Silva Martins

    Full Text Available INTRODUCTION: Persistence of cervical infection caused by human papillomavirus (HPV types with high oncogenic risk may lead to cervical intraepithelial neoplasia (CIN. The aim of the present study was to evaluate whether, in HIV-positive women, the presence of aneuploidy in cervical cell samples is associated with presence and evolution of CIN. METHODS: The present study had two stages. In the first stage, comprising a cross-sectional study, the association between the presence of aneuploidy seen via flow cytometry and sociodemographic characteristics, habits and characteristics relating to HPV and HIV infection was analyzed. In the second stage, comprising a cohort study, it was investigated whether aneuploidy was predictive of CIN evolution. RESULTS: No association was observed between the presence of aneuploidy and HPV infection, or between its presence and alterations seen in oncotic cytological analysis. On the other hand, aneuploidy was associated with the presence of CIN (p = 0.030 in histological analysis and with nonuse of antiretroviral therapy (p = 0.001. Most of the HIV-positive women (234/272 presented normal CD4+ T lymphocyte counts (greater than 350 cells/mm3 and showed a greater aneuploidy regression rate (77.5% than a progression rate (23.9% over a follow-up of up to two years. CONCLUSION: Although there was an association between the presence of cervical tissue lesions and the DNA index, the latter was not predictive of progression of the cervical lesion. This suggests that progression of the cervical lesion to cancer in HIV-positive women may also be changed through improvement of the immunological state enabled by using antiretroviral therapy.

  2. Isolation of HIV-1 RNA from plasma: evaluation of seven different methods for extraction (part two).

    Science.gov (United States)

    Fransen, K; Mortier, D; Heyndrickx, L; Verhofstede, C; Janssens, W; van der Groen, G

    1998-12-01

    Some new commercial methods for the extraction of viral RNA have been introduced recently. In addition to the study published previously (Verhofstede, C., Reniers, S., Van Wanzeele. F., Plum J., 1996. AIDS 8, 1421-1427), seven different methods (four newly developed and three reference methods) for extraction of HIV-1 RNA from plasma have been evaluated. The RNA preparation method that gave the best results (acceptable reproducibility, highest sensitivity, reasonable price, fast and easy to perform), was the QIAamp Viral RNA kit from QIAgen. The High Pure Viral RNA Kit (Boehringer Mannheim) as well as the non-commercialised extraction kits were also very sensitive. The non-commercial tests seem less suitable for routine use and for the processing of large number of samples. Two methods, RNA Insta-Pure LS (Eurogentec) and PANext RNA extraction kit 1 (NTL, PANsystems GmbH) are not adapted for HIV plasma extraction. The single step methods using glass fibre or silica column are rapid (from 60 to 75 min depending on the number of wash steps) and although the price is high they are cheaper than the Boom extraction methods: High Pure Viral RNA Kit (Boehringer Mannheim) ($3.3/sample), QIAamp Viral RNA Kit (Qiagen) ($3.6/sample), Boom extraction ($5/sample). The Qiagen kit is the only kit that combines sensitivity with reproducibility, it is commercialised, rapid and affordable in price and can be automated. For most of the methods evaluated the inter-test variability was acceptable (mean variation coefficient between duplicate extractions varied between 26.4 and 48.6%).

  3. Evaluation of school- and community-based HIV prevention interventions with junior secondary school students in Edo State, Nigeria.

    Science.gov (United States)

    Arnold, Robert; Maticka-Tyndale, Eleanor; Tenkorang, Eric; Holland, Daniel; Gaspard, Adeline; Luginaah, Isaac

    2012-06-01

    This study examined the impact of two interventions delivered in rural communities and schools in Edo State, Nigeria designed to decrease youth vulnerability to HIV infection. The Ministry of Education approved Family Life and HIV Education (FLHE) programme delivered in Junior Secondary Schools and a community-based initiative to raise AIDS Competency of rural communities were evaluated using a clustered randomized control trial and mixed qualitative-quantitative methods. Ten schools were assigned to each of three research arms: FLHE programme only, FLHE and community programme, and control. Results demonstrated positive effects on rejection of myths, attitudes related to abstinence and use of condoms, and sexual activity. Confidence in these results is supported by both levels of statistical significance and consistency in patterns of results across different levels of schooling. Results support expansion of delivery of the FLHE programme and development of community-based initiatives as effective methods of reducing youth vulnerability to HIV infection.

  4. Benchmarking HIV health care: from individual patient care to health care evaluation. An example from the EuroSIDA study.

    Science.gov (United States)

    Podlekareva, Daria N; Reekie, Joanne; Mocroft, Amanda; Losso, Marcelo; Rakhmanova, Aza G; Bakowska, Elzbieta; Karpov, Igor A; Lazarus, Jeffrey V; Gatell, Jose; Lundgren, Jens D; Kirk, Ole

    2012-09-25

    State-of-the-art care involving the utilisation of multiple health care interventions is the basis for an optimal long-term clinical prognosis for HIV-patients. We evaluated health care for HIV patients based on four key indicators. Four indicators of health care were assessed: Compliance with current guidelines on initiation of: 1) combination antiretroviral therapy (cART); 2) chemoprophylaxis; 3) frequency of laboratory monitoring; and 4) virological response to cART (proportion of patients with HIV-RNA 90% of time on cART). 7097 EuroSIDA patients were included from Northern (n = 923), Southern (n = 1059), West Central (n = 1290) East Central (n = 1366), Eastern (n = 1964) Europe, and Argentina (n = 495). Patients in Eastern Europe with a CD4 Argentina and Eastern Europe, respectively (p Argentina (adjusted OR 0.16 [95%CI 0.11-0.23, p < 0.0001]; 0.20[0.14-0.28, p < 0.0001] respectively). This assessment of HIV health care utilization revealed pronounced regional differences in adherence to guidelines and can help to identify gaps and direct target interventions. It may serve as a tool for the assessment and benchmarking of the clinical management of HIV patients in any setting worldwide.

  5. Priority setting in HIV/AIDS control in West Java Indonesia: an evaluation based on the accountability for reasonableness framework.

    Science.gov (United States)

    Tromp, Noor; Prawiranegara, Rozar; Subhan Riparev, Harris; Siregar, Adiatma; Sunjaya, Deni; Baltussen, Rob

    2015-04-01

    Indonesia has insufficient resources to adequately respond to the HIV/AIDS epidemic, and thus faces a great challenge in prioritizing interventions. In many countries, such priority setting processes are typically ad hoc and not transparent leading to unfair decisions. Here, we evaluated the priority setting process in HIV/AIDS control in West Java province against the four conditions of the accountability for reasonableness (A4R) framework: relevance, publicity, appeals and revision, and enforcement. We reviewed government documents and conducted semi-structured qualitative interviews based on the A4R framework with 22 participants of the 5-year HIV/AIDS strategy development for 2008-13 (West Java province) and 2007-11 (Bandung). We found that criteria for priority setting were used implicitly and that the strategies included a wide range of programmes. Many stakeholders were involved in the process but their contribution could be improved and particularly the public and people living with HIV/AIDS could be better engaged. The use of appeal and publicity mechanisms could be more transparent and formally stated. Public regulations are not yet installed to ensure fair priority setting. To increase fairness in HIV/AIDS priority setting, West Java should make improvements on all four conditions of the A4R framework. Published by Oxford University Press in association with The London School of Hygiene and Tropical Medicine © The Author 2014; all rights reserved.

  6. Evaluation of 'see-see and treat' strategy and role of HIV on cervical cancer prevention in Uganda

    Directory of Open Access Journals (Sweden)

    Sandin Sven

    2010-05-01

    Full Text Available Abstract Background There is scant information on whether Human Immunodeficiency Virus (HIV seropositivity has an influence on the outcome of treatment of precancerous cervical lesions using cryotherapy. We studied the prevalence of cervical abnormalities detectable by visual inspection and cervical lesions diagnosed by colposcopy according to HIV serostatus and described the outcomes of cryotherapy treatment. Methods Trained nurses examined women not previously screened for cervical cancer using visual inspection with acetic acid (VIA and Lugol's iodine (VILI in two family planning/post natal clinics in Kampala, Uganda, from February 2007 to August 2008. Women with abnormal visual inspection findings were referred for colposcopic evaluation and HIV testing. Women with precancerous cervical lesions detected at colposcopy were treated mainly by cryotherapy, and were evaluated for treatment outcome after 3 months by a second colposcopy. Results Of the 5 105 women screened, 834 presented a positive screening test and were referred for colposcopy. Of these 625 (75% returned for the colposcopic evaluation and were tested for HIV. For the 608 (97.5% women in the age range 20-60 years, colposcopy revealed 169 women with cervical lesions: 128 had inflammation, 19 had low grade squamous intraepithelial lesion (LGSIL, 13 had high grade squamous intraepithelial lesion (HGSIL, 9 had invasive cervical cancer and 2 had inconclusive findings. Detection rates per 1 000 women screened were higher among the older women (41-60 years compared to women aged 20-40 years. They were accordingly 55% and 20% for inflammation, 10% and 2% for LGSIL, 5% and 2% for HGSIL, 6% and 1% for invasive cervical cancer. Of the 608 women, 103 (16% were HIV positive. HIV positivity was associated with higher likelihood of inflammation (RR = 1.7; 95% CI: 1.2-2.4. Conclusions Detection rates were higher among older women 41-60 years. Visual inspection of the cervix uteri with acetic acid

  7. Mobile technology intervention to improve care coordination between HIV and substance use treatment providers: development, training, and evaluation protocol.

    Science.gov (United States)

    Claborn, Kasey; Becker, Sara; Ramsey, Susan; Rich, Josiah; Friedmann, Peter D

    2017-03-14

    People living with HIV (PLWH) with a substance use disorder (SUD) tend to receive inadequate medical care in part because of a siloed healthcare system in which HIV and substance use services are delivered separately. Ideal treatment requires an interdisciplinary, team-based coordinated care approach, but many structural and systemic barriers impede the integration of HIV and SUD services. The current protocol describes the development and preliminary evaluation of a care coordination intervention (CCI), consisting of a tablet-based mobile platform for HIV and SUD treatment providers, an interagency communication protocol, and a training protocol. We hypothesize that HIV and SUD treatment providers will find the CCI to be acceptable, and that after receipt of the CCI, providers will: exhibit higher retention in dual care among patients, report increased frequency and quality of communication, and report increased rates of relational coordination. A three phase approach is used to refine and evaluate the CCI. Phase 1 consists of in-depth qualitative interviews with 8 key stakeholders as well as clinical audits of participating HIV and SUD treatment agencies. Phase 2 contains functionality testing of the mobile platform with frontline HIV and SUD treatment providers, followed by refinement of the CCI. Phase 3 consists of a pre-, post-test trial with 30 SUD and 30 HIV treatment providers. Data will be collected at the provider, organization, and patient levels. Providers will complete assessments at baseline, immediately post-training, and at 1-, 3-, and 6-months post-training. Organizational data will be collected at baseline, 1-, 3-, and 6-months post training, while patient data will be collected at baseline and 6-months post training. This study will develop and evaluate a CCI consisting of a tablet-based mobile platform for treatment providers, an interagency communication protocol, and a training protocol as a means of improving the integration of care for PLWH

  8. Efficacy and safety of etravirine in treatment-experienced, HIV-1 patients: pooled 48 week analysis of two randomized, controlled trials

    NARCIS (Netherlands)

    Katlama, Christine; Haubrich, Richard; Lalezari, Jacob; Lazzarin, Adriano; Madruga, José V.; Molina, Jean-Michel; Schechter, Mauro; Peeters, Monika; Picchio, Gaston; Vingerhoets, Johan; Woodfall, Brian; de Smedt, Goedele; Ariza, H. A.; Benetucci, J.; Cahn, P.; Calanni, L. M.; Cassetti, I.; Corral, J.; David, D. O.; Krolewiecki, A.; Losso, M. H.; Patterson, P.; Teijeiro, R. A.; Grinsztejn, B.; da Cunha, C. A.; Kallas, E. G.; Netto, E. M.; Pilotto, J. H.; Suleiman, J.; Timerman, A.; Ballesteros, J.; Northland, R.; Alvilés Montoya, A. A.; Herrera Martinez, G.; Solano Chinchilla, A.; Dupon, M.; Livrozet, J. M.; Morlat, P.; Pialoux, G.; Piketty, C.; Poizot-Martin, I.; Andrade-Villanueva, J.; Reyes-Terán, G.; Sierra-Madero, J.; Canton, A.; Rodriguez, A.; Sosa, N.; Morales Ramirez, J. O.; Santana Bagur, J. L.; Soto-Malave, R.; Anekthananon, T.; Mootsikapun, P.; Ruxrungtham, K.; Albrecht, M.; Bellos, N.; Bolan, R.; Brachman, P.; Brinson, C.; Cruickshank, F.; Elion, R.; Fessel, W. J.; Hawkins, T.; Hodder, S.; Hutcherson, P.; Jefferson, T.; Katner, H.; Kinder, C.; Kozal, M.; Leider, J.; Mills, T.; McDonough, D.; Mounzer, K.; Nadler, J.; Norris, D.; O'Brien, W.; Pierone, G.; Raben, K.; Rashbaum, B.; Rawlings, M.; Rodwick, B.; Ruane, P.; Sampson, J.; Schrader, S.; Scribner, A.; Sension, M.; Sweet, D.; Wade, B.; Wheeler, D.; Wilkin, A.; Wills, T.; Wohlfeiler, M.; Workowski, K.; Chuah, J.; Cooper, D.; Eu, B.; Hoy, J.; Workman, C.; Clumeck, N.; Colebunders, R.; Moutschen, M.; Gill, J.; Gough, K.; Junod, P.; Kilby, D.; Montaner, J.; Rachlis, A.; Trottier, B.; Tsoukas, C. M.; Walmsley, S. L.; Arvieux, C.; Cotte, L.; Delfraissy, J. F.; Girard, P. M.; Marchou, B.; Vittecoq, D.; Yazdanpanah, Y.; Yeni, P.; Arastéh, K.; Esser, S.; Fätkenheuer, G.; Gellermann, H.; Göbels, K.; Goebel, F. D.; Jäger, H.; Moll, A.; Rockstroh, J. K.; Schuster, D.; Staszewski, S.; Stoehr, A.; Antinori, A.; Carosi, G.; Di Perri, G.; Esposito, R.; Mazzotta, F.; Pagano, G.; Raise, E.; Rusconi, S.; Sighinolfi, L.; Suter, F.; Frissen, P. H. J.; Prins, J. M.; Rijnders, B. J. A.; Horban, A.; Antunes, F.; Miranda, M.; Vera, J.; Clotet, B.; Domingo, P.; Garcia, G.; Gatell, J. M.; González-Lahoz, J.; López-Aldeguer, J.; Podzamczer, D.; Easterbrook, P.; Fisher, M.; Johnson, M.; Orkin, C.; Wilkins, E.; Barnett, B.; Baxter, J.; Beatty, G.; Berger, D.; Borkert, C.; Campell, T.; Cohen, C.; Conant, M.; Ernst, J.; Farthing, C.; File, T.; Frank, M.; Gallant, J. E.; Greenberg, R. N.; Hicks, C.; Jayaweera, D. T.; Kerkar, S.; Markowitz, N.; Martorell, C.; McDonald, C.; McMahon, D.; Mogyoros, M.; Myers, R. A.; Richmond, G.; Sathasivam, K.; Schneider, S.; Schrager, H.; Shalit, P.; Siegal, F. P.; Sloan, L.; Smith, K.; Smith, S.; Tebas, P.; Tkatch, L. S.; Towner, W.

    2009-01-01

    OBJECTIVE: To evaluate the efficacy, safety and virologic resistance profile of etravirine (TMC125), a next-generation nonnucleoside reverse transcriptase inhibitor, over 48 weeks in treatment-experienced adults infected with HIV-1 strains resistant to a nonnucleoside reverse transcriptase inhibitor

  9. HIV testing in Europe: Evaluating the impact, added value, relevance and usability of the European Centre for Disease Prevention and Control (ECDC)'s 2010 HIV testing guidance.

    Science.gov (United States)

    Sullivan, Ann K; Sperle, Ida; Raben, Dorthe; Amato-Gauci, Andrew J; Lundgren, Jens Dilling; Yazdanpanah, Yazdan; Jakobsen, Stine Finne; Tavoschi, Lara

    2017-11-01

    An evaluation of the 2010 ECDC guidance on HIV testing, conducted in October 2015-January 2016, assessed its impact, added value, relevance and usability and the need for updated guidance. Data sources were two surveys: one for the primary target audience (health policymakers and decision makers, national programme managers and ECDC official contact points in the European Union/European Economic Area (EU/EEA) countries and one for a broader target audience (clinicians, civil society organisations and international public health agencies); two moderated focus group discussions  (17 participants each); webpage access data; a literature citation review; and an expert consultation (18 participants) to discuss the evaluation findings. Twenty-three of 28 primary target audience and 31 of 51 broader target audience respondents indicated the guidance was the most relevant when compared with other international guidance. Primary target audience respondents in 11 of 23 countries reported that they had used the guidance in development, monitoring and/or evaluation of their national HIV testing policy, guidelines, programme and/or strategy, and 29 of 51 of the broader target audience respondents reported having used the guidance in their work. Both the primary and broader target audience considered it important or very important to have an EU/EEA-level HIV testing guidance (23/28 and 46/51, respectively). The guidance has been widely used to develop policies, guidelines, programmes and strategies in the EU/EEA and should be regularly updated due to continuous developments in the field in order to continue to serve as an important reference guidance in the region.

  10. A human rights-focused HIV intervention for sex workers in Metro Manila, Philippines: evaluation of effects in a quantitative pilot study.

    Science.gov (United States)

    Urada, Lianne A; Simmons, Janie; Wong, Betty; Tsuyuki, Kiyomi; Condino-Enrera, Gerlita; Hernandez, Laufred I; Simbulan, Nymia Pimentel; Raj, Anita

    2016-11-01

    This study evaluated a brief human rights-focused HIV community mobilization intervention for sex workers in the Philippines, a country with one of the fastest rising number of HIV cases worldwide. Five single-session group interventions to reduce sexual risk and increase HIV testing among 86 sex workers in Manila were evaluated with pre-post-test data via Wilcoxon's signed-ranks and Mann-Whitney tests. The 4-h intervention, Kapihan (August-November, 2013), integrated human rights with HIV skill-building. Demographic data, violence/trafficking victimization, human rights knowledge, and intentions to HIV test and treat were collected. Participants were median aged 23; female (69 %); had children (55; 22 % had 3+ children); used drugs (past 3 months: 16 %); sexually/physically abused by clients (66 %); 20 % street sex workers ever took an HIV test. Pre-post-test scores significantly improved in knowledge of HIV (z = -8.895, p human rights (z = -4.391, p rights of research participants (z = -5.081, p human rights into HIV interventions may empower sex workers to address their health and human rights and test for HIV.

  11. Programmatic evaluation of a combined antigen and antibody test for rapid HIV diagnosis in a community and sexual health clinic screening programme.

    Science.gov (United States)

    Taegtmeyer, Miriam; MacPherson, Peter; Jones, Kathy; Hopkins, Mark; Moorcroft, Jay; Lalloo, David G; Chawla, Anu

    2011-01-01

    A substantial proportion of HIV-infected individuals in the UK are unaware of their status and late presentations continue, especially in low prevalence areas. Fourth generation antigen/antibody rapid test kits could facilitate earlier diagnosis of HIV in non-clinical settings but lack data on performance under programmatic conditions. We evaluated the performance of Determine HIV-1/2 Ag/Ab Combo Test (Determine Combo), a rapid test with indicators for both HIV antibodies and p24 antigen, in participants recruited from community outreach and hospital-based sexual health clinics. HIV infection was confirmed using laboratory enzyme-linked immunosorbent assay (EIA), Line Immuno Assay (LIA) and quantitative polymerase chain reaction (PCR). In total, 953 people underwent HIV testing. HIV antibody (Ab) prevalence was 1.8% (17/953). Four false positive rapid tests were identified: two antibody and two p24 antigen (Ag) reactions. Of participants diagnosed as HIV Ab positive, 2/17 (12%) were recent seroconverters based on clinical history and HIV antibody avidity test results. However, none of these were detected by the p24 antigen component of the rapid test kit. There were no other true positive p24 Ag tests. These data lend support to an increasing body of evidence suggesting that 4th generation rapid HIV tests have little additional benefit over 3rd generation HIV kits for routine screening in low prevalence settings and have high rates of false positives. In order to optimally combine community-based case-finding among hard-to-reach groups with reliable and early diagnosis 3rd generation kits should be primarily used with laboratory testing of individuals thought to be at risk of acute HIV infection. A more reliable point of care diagnostic is required for the accurate detection of acute HIV infection under programmatic conditions.

  12. Programmatic evaluation of a combined antigen and antibody test for rapid HIV diagnosis in a community and sexual health clinic screening programme.

    Directory of Open Access Journals (Sweden)

    Miriam Taegtmeyer

    Full Text Available BACKGROUND: A substantial proportion of HIV-infected individuals in the UK are unaware of their status and late presentations continue, especially in low prevalence areas. Fourth generation antigen/antibody rapid test kits could facilitate earlier diagnosis of HIV in non-clinical settings but lack data on performance under programmatic conditions. METHODS AND FINDINGS: We evaluated the performance of Determine HIV-1/2 Ag/Ab Combo Test (Determine Combo, a rapid test with indicators for both HIV antibodies and p24 antigen, in participants recruited from community outreach and hospital-based sexual health clinics. HIV infection was confirmed using laboratory enzyme-linked immunosorbent assay (EIA, Line Immuno Assay (LIA and quantitative polymerase chain reaction (PCR. In total, 953 people underwent HIV testing. HIV antibody (Ab prevalence was 1.8% (17/953. Four false positive rapid tests were identified: two antibody and two p24 antigen (Ag reactions. Of participants diagnosed as HIV Ab positive, 2/17 (12% were recent seroconverters based on clinical history and HIV antibody avidity test results. However, none of these were detected by the p24 antigen component of the rapid test kit. There were no other true positive p24 Ag tests. CONCLUSION: These data lend support to an increasing body of evidence suggesting that 4th generation rapid HIV tests have little additional benefit over 3rd generation HIV kits for routine screening in low prevalence settings and have high rates of false positives. In order to optimally combine community-based case-finding among hard-to-reach groups with reliable and early diagnosis 3rd generation kits should be primarily used with laboratory testing of individuals thought to be at risk of acute HIV infection. A more reliable point of care diagnostic is required for the accurate detection of acute HIV infection under programmatic conditions.

  13. Evaluating the cost-effectiveness of combination antiretroviral therapy for the prevention of mother-to-child transmission of HIV in Uganda

    NARCIS (Netherlands)

    Kuznik, Andreas; Lamorde, Mohammed; Hermans, Sabine; Castelnuovo, Barbara; Auerbach, Brandon; Semeere, Aggrey; Sempa, Joseph; Ssennono, Mark; Ssewankambo, Fred; Manabe, Yukari C.

    2012-01-01

    Objective To model the cost-effectiveness in Uganda of combination antiretroviral therapy (ART) to prevent mother-to-child transmission of human immunodeficiency virus (HIV). Methods The cost-effectiveness of ART was evaluated on the assumption that ART reduces the risk of an HIV-positive pregnant

  14. Evaluation of emotion processing in HIV-infected patients and correlation with cognitive performance

    OpenAIRE

    Baldonero, Eleonora; Ciccarelli, Nicoletta; Fabbiani, Massimiliano; Colafigli, Manuela; Improta, Erika; D?Avino, Alessandro; Mondi, Annalisa; Cauda, Roberto; Di Giambenedetto, Simona; Silveri, Maria Caterina

    2013-01-01

    Background Facial emotion recognition depends on cortical and subcortical networks. HIV infection of the central nervous system can damage these networks, leading to impaired facial emotion recognition. Methods We performed a cross-sectional single cohort study consecutively enrolling HIV?+?subjects during routine outpatient visits. Age, gender and education-matched HIV-negative healthy individuals were also selected. Subjects were submitted to a Facial Emotion Recognition Test, which assesse...

  15. First-line HIV treatment: evaluation of backbone choice and its budget impact

    Directory of Open Access Journals (Sweden)

    Orietta Zaniolo

    2013-06-01

    Full Text Available OBJECTIVE: The gradual increase of persons living with HIV, mainly due to the reduced mortality achieved with effective antiretroviral therapies, calls for increased rationality and awareness in health resources consumption also during the early illness phases. Aim of this work is the estimation of the budget impact related to the variation in backbone prescribing trends in naïve patients.METHODS: Target population is the number of patients starting antiretroviral therapy each year, according to the Italian HIV surveillance registry, excluding patients receiving non-authorized or non-recommended regimens. We modeled 3-year mortality and durability rates on a dynamic cohort, basing on international literature. A prevalent patients analysis has also been conducted, for which the model is fed by a closed cohort consisting of all the patients without experience of virologic failure. The aim of this collateral analysis is to estimate the difference in current annual expenditures if the past prescription trends for patients starting therapy would have led to the evaluated hypothetical scenarios. Current Italian market shares of triple regimens containing first-choice or alternative backbones (tenofovir/emtricitabine, abacavir/lamivudine, tenofovir/lamivudine and zidovudine/lamivudine are compared to three hypothetical scenarios (base-case, minimum and maximum in which increasing shares of patients eligible to abacavir/lamivudine start first line treatment with this backbone. Annual cost for each regimen comprises drugs acquisition under hospital pricing rules, monitoring exams and preventive tests, valued basing on regional reimbursement tariffs.RESULTS: According to current prescribing trends, in the next three years about 13,000 patients starting HIV therapy will receive tenofovir/emtricitabine (83% of the target population, and minor portions other regimens (9% abacavir/lamivudine, 8% zidovudine/lamivudine. Patients that would be eligible to

  16. Syphilis detection: evaluation of serological screening and pilot reverse confirmatory assay algorithm in blood donors.

    Science.gov (United States)

    Sommese, Linda; Paolillo, Rossella; Sabia, Chiara; Costa, Dario; De Pascale, Maria Rosaria; Iannone, Carmela; Esposito, Antonella; Schiano, Concetta; Napoli, Claudio

    2016-07-01

    Serological assays are still considered the most useful tests in the diagnosis of syphilis. Since no single serological assay is able to provide a satisfactory result, in our laboratory we have evaluated the usefulness of a commercially-available immunoblot to diagnose syphilis infection among blood donors. From October 2012 to June 2013, 4572 blood donors were screened for syphilis with an automated chemiluminescent microparticle immunoassay (CMIA). To confirm the presence of treponemal antibodies, CMIA-reactive sera were tested by standard Treponema pallidum haemagglutination assay (TPHA). In addition, an alternative confirmatory test - the immunoblot INNO-LIA assay was introduced in our laboratory. Since two additional positives among CMIA-reactive-TPHA-negative samples were found, we concluded that the INNO-LIA immunoblot allowed a better detection of syphilis compared to TPHA. A confirmatory strategy based on the use of two treponemal assays could meet the screening requirements for blood donors as well as in our centre. © The Author(s) 2015.

  17. HIV: Treatment and Comorbidity

    NARCIS (Netherlands)

    C. Rokx (Casper)

    2016-01-01

    markdownabstractClinicians worldwide strive to improve HIV care for their patients. Antiretroviral therapy prevents HIV related mortality and is lifelong. A clinical evaluation of these treatment strategies is necessary to identify strategies that may jeopardize treatment effectiveness and patient

  18. Evaluation of the treatment of reverse osmosis concentrates from municipal wastewater reclamation by coagulation and granular activated carbon adsorption.

    Science.gov (United States)

    Sun, Ying-Xue; Yang, Zhe; Ye, Tao; Shi, Na; Tian, Yuan

    2016-07-01

    Reverse osmosis concentrate (ROC) from municipal wastewater reclamation reverse osmosis (mWRRO) contains elevated concentrations of contaminants which pose potential risks to aquatic environment. The treatment of ROC from an mWRRO using granular activated carbon (GAC) combined pretreatment of coagulation was optimized and evaluated. Among the three coagulants tested, ferric chloride (FeCl3) presented relatively higher DOC removal efficiency than polyaluminium chloride and lime at the same dosage and coagulation conditions. The removal efficiency of DOC, genotoxicity, and antiestrogenic activity concentration of the ROC could achieve 16.9, 18.9, and 39.7 %, respectively, by FeCl3 coagulation (with FeCl3 dosage of 180.22 mg/L), which can hardly reduce UV254 and genotoxicity normalized by DOC of the DOM with MW <5 kDa. However, the post-GAC adsorption column (with filtration velocity of 5.7 m/h, breakthrough point adsorption capacity of 0.22 mg DOC/g GAC) exhibited excellent removal efficiency on the dominant DOM fraction of MW <5 kDa in the ROC. The removal efficiency of DOC, UV254, and TDS in the ROC was up to 91.8, 96, and 76.5 %, respectively, by the FeCl3 coagulation and post-GAC adsorption. Also, the DOM with both genotoxicity and antiestrogenic activity were completely eliminated by the GAC adsorption. The results suggest that GAC adsorption combined pretreatment of FeCl3 coagulation as an efficient method to control organics, genotoxicity, and antiestrogenic activity in the ROC from mWRRO system.

  19. Expansion of CD56- NK cells in chronic HCV/HIV-1 co-infection: reversion by antiviral treatment with pegylated IFNalpha and ribavirin.

    Science.gov (United States)

    Gonzalez, Veronica D; Falconer, Karolin; Michaëlsson, Jakob; Moll, Markus; Reichard, Olle; Alaeus, Annette; Sandberg, Johan K

    2008-07-01

    Co-infection with HCV and HIV-1 is a problem of increasing importance and the role of innate cellular immunity in this co-infection is incompletely understood. Here, we have observed sharply elevated numbers of CD56(-)CD16(+) perforin(low) NK cells in HCV/HIV-1 co-infected subjects on antiretroviral therapy. Interestingly, this expansion of unconventional CD56(-) NK cells rapidly reverted when HCV was suppressed by IFNalpha and ribavirin treatment, and was not seen in mono-infected control groups. In vitro experiments suggested that this effect of treatment was due to suppression of HCV viremia rather than a direct effect of IFNalpha on these cells. In contrast, the conventional CD56(+) NK cells were largely unchanged in subjects with high HCV loads, although they exhibited slightly decreased perforin expression. With delayed kinetics, the CD56(bright) immuno-regulatory NK cell subset temporarily increased to supranormal levels in response to HCV treatment. In contrast to the NK compartment, the CD1d-restricted NKT cells were severely reduced by the co-infection and not restored by treatment. Together, our data suggest that the high HCV loads in HCV/HIV-1 co-infection alter the NK cell compartment in a way not observed in HCV mono-infection.

  20. The K65R mutant reverse transcriptase of HIV-1 cross-resistant to 2', 3'-dideoxycytidine, 2',3'-dideoxy-3'-thiacytidine, and 2',3'-dideoxyinosine shows reduced sensitivity to specific dideoxynucleoside triphosphate inhibitors in vitro.

    Science.gov (United States)

    Gu, Z; Fletcher, R S; Arts, E J; Wainberg, M A; Parniak, M A

    1994-11-11

    The K65R mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) encodes cross-resistance to 2',3'-dideoxycytidine (ddC), 2',3'-dideoxy-3'-thiacytidine (3TC), and 2',3'-dideoxyinosine (ddI). We characterized the in vitro sensitivities of recombinant wild type (wt) and K65R mutant RT to dideoxynucleoside triphosphate (ddNTP) inhibitors, using a variety of primer-templates. With poly(rA)-oligo(dT), the K65R mutant showed slight increases in Ki for ddTTP and 3'-azido, 3'-deoxythymidine triphosphate (AZTTP) compared to wt RT, but neither wt nor K65R RT was inhibited by ddCTP or ddATP. With poly(rI)-oligo(dC), the K65R mutant showed a 2-fold increase in Km for dCTP and a 20-fold increase in Ki for ddCTP compared to wt, whereas ddATP, ddTTP, and AZTTP failed to inhibit either enzyme. With a heteropolymeric primer-template, the K65R mutant showed 10-fold reduced sensitivities to ddCTP, 3TCTP, and ddATP, and 4-fold reduced sensitivity to AZTTP, compared to wt. In contrast, both enzymes were equally inhibited by ddTTP and ddGTP. HIV-1 cross-resistance to ddC/3TC/ddI resulting from the K65R mutation may therefore involve selective alterations in substrate/inhibitor recognition. Additionally, competitive inhibition by ddNTPs noncomplementary to the template base appears to be unimportant in the mechanism of inhibition of HIV-1 RT by dideoxynucleoside analogs.

  1. An Outdated Notion of Antibody Specificity is One of the Major Detrimental Assumptions of the Structure-Based Reverse Vaccinology Paradigm, Which Prevented It from Helping to Develop an Effective HIV-1 Vaccine

    Science.gov (United States)

    Van Regenmortel, Marc H. V.

    2014-01-01

    The importance of paradigms for guiding scientific research is explained with reference to the seminal work of Karl Popper and Thomas Kuhn. A prevalent paradigm, followed for more than a decade in HIV-1 vaccine research, which gave rise to the strategy known as structure-based reverse vaccinology is described in detail. Several reasons why this paradigm did not allow the development of an effective HIV-1 vaccine are analyzed. A major reason is the belief shared by many vaccinologists that antibodies possess a narrow specificity for a single epitope and are not polyspecific for a diverse group of potential epitopes. When this belief is abandoned, it becomes obvious that the one particular epitope structure observed during the crystallographic analysis of a neutralizing antibody–antigen complex does not necessarily reveal, which immunogenic structure should be used to elicit the same type of neutralizing antibody. In the physical sciences, scientific explanations are usually presented as logical deductions derived from a relevant law of nature together with certain initial conditions. In immunology, causal explanations in terms of a single cause acting according to a law of nature are not possible because numerous factors always play a role in bringing about an effect. The implications of this state of affairs for the rational design of HIV vaccines are outlined. An alternative approach to obtain useful scientific understanding consists in intervening empirically in the immune system and it is suggested that manipulating the system experimentally is needed to learn to control it and achieve protective immunity by vaccination. PMID:25477882

  2. An outdated notion of antibody specificity is one of the major detrimental assumptions of the structure-based reverse vaccinology paradigm which prevented it from helping to develop an effective HIV-1 vaccine

    Directory of Open Access Journals (Sweden)

    Marc H V Van Regenmortel

    2014-11-01

    Full Text Available The importance of paradigms for guiding scientific research is explained with reference to the seminal work of Karl Popper and Thomas Kuhn. A prevalent paradigm, followed for more than a decade in HIV-1 vaccine research, which gave rise to the strategy known as structure-based reverse vaccinology is described in detail. Several reasons why this paradigm did not allow the development of an effective HIV-1 vaccine are analyzed. A major reason is the belief shared by many vaccinologists that antibodies possess a narrow specificity for a single epitope and are not polyspecific for a diverse group of potential epitopes. When this belief is abandoned, it becomes obvious that the one particular epitope structure observed during the crystallographic analysis of a neutralizing antibody-antigen complex does not necessarily reveal which immunogenic structure should be used to elicit the same type of neutralizing antibody.In the physical sciences, scientific explanations are usually presented as logical deductions derived from a relevant law of nature together with certain initial conditions. In immunology, causal explanations in terms of a single cause acting according to a law of nature are not possible because numerous factors always play a role in bringing about an effect. The implications of this state of affairs for the rational design of HIV vaccines are outlined. An alternative approach to obtain useful scientific understanding consists in intervening empirically in the immune system and it is suggested that manipulating the system experimentally is needed to learn to control it and achieve protective immunity by vaccination.

  3. Evaluation of four commercial virological assays for early infant HIV-1 diagnosis using dried blood specimens.

    Science.gov (United States)

    Alvarez, Patricia; Prieto, Luis; Martín, Leticia; Obiang, Jacinta; Avedillo, Pedro; Vargas, Antonio; Rojo, Pablo; Fernández McPhee, Carolina; Sanz Canalejas, Leticia; Benito, Agustín; Ramos, José Tomás; Holguín, África

    2017-01-01

    Early infant diagnosis (EID) of HIV-1 is necessary to reduce HIV-related mortality. As maternal antibodies transferred across the placenta may persist up to 18 mo, commercial virological assays (CVAs) are needed. This study compares four CVAs for EID using dried blood specimens (DBS) from HIV-1-exposed infants. DBS from 68 infants born to HIV-1-infected women were collected from November 2012 to December 2013 in Equatorial Guinea. Four CVAs were performed: Siemens VERSANT HIV-1 RNA 1.0 kPCR assay, Roche CAP/CTM Quantitative Test v2.0, CAP/CTM Qualitative Tests v1.0 and v2.0. Definitive diagnosis was established following World Health Organization (WHO) recommendations. Two HIV-1-infected infants (2.9%) were detected by the four CVAs while 49 (72%) resulted negative. Discordant resu