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Sample records for simple rapid hiv

  1. Evaluation of five simple rapid HIV assays for potential use in the Brazilian national HIV testing algorithm.

    PubMed

    da Motta, Leonardo Rapone; Vanni, Andréa Cristina; Kato, Sérgio Kakuta; Borges, Luiz Gustavo dos Anjos; Sperhacke, Rosa Dea; Ribeiro, Rosangela Maria M; Inocêncio, Lilian Amaral

    2013-12-01

    Since 2005, the Department of Sexually Transmitted Diseases (STDs), Acquired Immunodeficiency Syndrome (AIDS) and Viral Hepatitis under the Health Surveillance Secretariat in Brazil's Ministry of Health has approved a testing algorithm for using rapid human immunodeficiency virus (HIV) tests in the country. Given the constant emergence of new rapid HIV tests in the market, it is necessary to maintain an evaluation program for them. Conscious of this need, this multicenter study was conducted to evaluate five commercially available rapid HIV tests used to detect anti-HIV antibodies in Brazil. The five commercial rapid tests under assessment were the VIKIA HIV-1/2 (bioMérieux, Rio de Janeiro, Brazil), the Rapid Check HIV 1 & 2 (Center of Infectious Diseases, Federal University of Espírito Santo, Vitória, Brazil), the HIV-1/2 3.0 Strip Test Bioeasy (S.D., Kyonggi-do, South Korea), the Labtest HIV (Labtest Diagnóstica, Lagoa Santa, Brazil) and the HIV-1/2 Rapid Test Bio-Manguinhos (Oswaldo Cruz Foundation, Rio de Janeiro, Brazil). A total of 972 whole-blood samples were collected from HIV-infected patients, pregnant women and individuals seeking voluntary counselling and testing who were recruited from five centers in different regions of the country. Informed consent was obtained from the study participants. The results were compared with those obtained using the HIV algorithm used currently in Brazil, which includes two enzyme immunoassays and one Western blot test. The operational performance of each assay was also compared to the defined criteria. A total of 972 samples were tested using reference assays, and the results indicated 143 (14.7%) reactive samples and 829 (85.3%) nonreactive samples. Sensitivity values ranged from 99.3 to 100%, and specificity was 100% for all five rapid tests. All of the rapid tests performed well, were easy to perform and yielded high scores in the operational performance analysis. Three tests, however, fulfilled all of the

  2. Rapid and simple screening and supplemental testing for HIV-1 and HIV-2 infections in west Africa.

    PubMed

    Brattegaard, K; Kouadio, J; Adom, M L; Doorly, R; George, J R; De Cock, K M

    1993-06-01

    Researchers from an AIDS research project took blood samples from 1000 consecutive women during childbirth at a maternal and child health center in Abidjan, Cote d'Ivoire, and from 185 hospitalized patients to compare the results of a combination of synthetic peptide-based rapid tests (product names, Testpack and Genie), which check for HIV-1 and HIV-2 antibodies, with those of the Western Blot-based test. They also wanted to see whether the rapid test-based strategy could replace the Western Blot-based test as a supplemental test. The Western Blot indicated the HIV-1 and/or HIV-2 prevalence to be 13% among the new mothers and 78% among the hospitalized patients for an overall prevalence of 23%. 3.3% of all people were positive for both HIV-1 and HIV-2. 17.4% tested positive for just HIV-1. 2.1% were positive for HIV-2. The rapid tests had a sensitivity of 99.6% and a specificity of 99.9%. The positive predictive value was 99.6% and the negative predictive value was 99.9%. The rapid tests identified 4% of the HIV-2 positive samples and 1% of the HIV-1 samples to be dually reactive. These findings demonstrated that rapid synthetic peptide-based assays reliably detect HIV-1 and HIV-2 antibodies and can be supplemental tests. High quality HIV serology can be performed in a setting without running water and electricity which was the case in this study. A further advantage of this strategy is that each test takes only 10 minutes. These tests would have significant effects on HIV testing and counseling, diagnosis, and screening of blood for transfusion in rural areas of developing countries. PMID:8395857

  3. Convenient cell fusion assay for rapid screening for HIV entry inhibitors

    NASA Astrophysics Data System (ADS)

    Jiang, Shibo; Radigan, Lin; Zhang, Li

    2000-03-01

    Human immunodeficiency viruses (HIV)-induced cell fusion is a critical pathway of HIV spread from infected cells to uninfected cells. A rapid and simple assay was established to measure HIV-induce cell fusion. This study is particularly useful to rapid screen for HIV inhibitors that block HIV cell-to-cell transmission. Present study demonstrated that coculture of HIV-infected cells with uninfected cells at 37 degree(s)C for 2 hours resulted in the highest cell fusion rate. Using this cell fusion assay, we have identified several potent HIV inhibitors targeted to the HIV gp41 core. These antiviral agents can be potentially developed as antiviral drugs for chemotherapy and prophylaxis of HIV infection and AIDS.

  4. A simple, rapid, and sensitive system for the evaluation of anti-viral drugs in rats

    SciTech Connect

    Li, Xiaoguang; Qian, Hua; Miyamoto, Fusako; Kawaji, Kumi; Hattori, Toshio; Watanabe, Kentaro; Oishi, Shinya; Fujii, Nobutaka; and others

    2012-07-27

    Highlights: Black-Right-Pointing-Pointer We established a novel, simple and rapid in vivo system for evaluation of anti-HIV-1 drugs with rats. Black-Right-Pointing-Pointer The system may be applicable for other antiviral drugs, and/or useful for initial screening in vivo. Black-Right-Pointing-Pointer In this system, TRI-1144 displayed the most potent anti-HIV-1 activity in vivo. -- Abstract: The lack of small animal models for the evaluation of anti-human immunodeficiency virus type 1 (HIV-1) agents hampers drug development. Here, we describe the establishment of a simple and rapid evaluation system in a rat model without animal infection facilities. After intraperitoneal administration of test drugs to rats, antiviral activity in the sera was examined by the MAGI assay. Recently developed inhibitors for HIV-1 entry, two CXCR4 antagonists, TF14016 and FC131, and four fusion inhibitors, T-20, T-20EK, SC29EK, and TRI-1144, were evaluated using HIV-1{sub IIIB} and HIV-1{sub BaL} as representative CXCR4- and CCR5-tropic HIV-1 strains, respectively. CXCR4 antagonists were shown to only possess anti-HIV-1{sub IIIB} activity, whereas fusion inhibitors showed both anti-HIV-1{sub IIIB} and anti-HIV-1{sub BaL} activities in rat sera. These results indicate that test drugs were successfully processed into the rat sera and could be detected by the MAGI assay. In this system, TRI-1144 showed the most potent and sustained antiviral activity. Sera from animals not administered drugs showed substantial anti-HIV-1 activity, indicating that relatively high dose or activity of the test drugs might be needed. In conclusion, the novel rat system established here, 'phenotypic drug evaluation', may be applicable for the evaluation of various antiviral drugs in vivo.

  5. Simple rapid method for gene transfer

    SciTech Connect

    Cockburn, A.F.; Meier, H.

    1990-01-30

    The object of the present invention is to provide methods for gene transfer that reduce or eliminate cellular pretreatment steps, e.g., the removal of cell wall by chemical or enzymatic methods, is rapid and can be practiced without the need of additional expensive equipment. Cells, embryos or tissues selected for genetic manipulation are suspended in an Eppendorf tube in an aliquot of the desired genetic material to be transferred to which the resulting mixture is added and is agitated by vortexing from about 30 to about 90 seconds. The cells, embryos or tissue are sedimented and the DNA supernatant removed. After sedimentation, the injected material is resuspended in or on a growth medium to assay for expression.

  6. Determination of HIV status in African adults with discordant HIV rapid tests

    PubMed Central

    Fogel, Jessica M.; Piwowar-Manning, Estelle; Donohue, Kelsey; Cummings, Vanessa; Marzinke, Mark A.; Clarke, William; Breaud, Autumn; Fiamma, Agnès; Donnell, Deborah; Kulich, Michal; Mbwambo, Jessie K. K.; Richter, Linda; Gray, Glenda; Sweat, Michael; Coates, Thomas J.; Eshleman, Susan H.

    2015-01-01

    Background In resource-limited settings, HIV infection is often diagnosed using two rapid tests. If the results are discordant, a third tie-breaker test is often used to determine HIV status. This study characterized samples with discordant rapid tests and compared different testing strategies for determining HIV status in these cases. Methods Samples were previously collected from 173 African adults in a population-based survey who had discordant rapid test results. Samples were classified as HIV positive or HIV negative using a rigorous testing algorithm that included two fourth-generation tests, a discriminatory test, and two HIV RNA tests. Tie-breaker tests were evaluated, including: rapid tests (one performed in-country), a third-generation enzyme immunoassay (EIA), and two fourth-generation tests. Selected samples were further characterized using additional assays. Results Twenty-nine (16.8%) samples were classified as HIV positive; 24 (82.8%) of those samples had undetectable HIV RNA. Antiretroviral drugs were detected in one sample. Sensitivity was 8.3%–43% for the rapid tests; 24.1% for the third-generation EIA; 95.8% and 96.6% for the fourth-generation tests. Specificity was lower for the fourth-generation tests than the other tests. Accuracy ranged from 79.5–91.3%. Conclusions In this population-based survey, most HIV-infected adults with discordant rapid tests were virally suppressed without antiretroviral drugs. Use of individual assays as tie-breaker tests was not a reliable method for determining HIV status in these individuals. More extensive testing algorithms that use a fourth-generation screening test with a discriminatory test and HIV RNA test are preferable for determining HIV status in these cases. PMID:25835607

  7. Clinical performance of the Multispot HIV-1/HIV-2 rapid test to correctly differentiate HIV-2 from HIV-1 infection in screening algorithms using third and fourth generation assays and to identify cross reactivity with the HIV-1 Western Blot

    PubMed Central

    Ramos, Eric M.; Harb, Socorro; Dragavon, Joan; Coombs, Robert W.

    2014-01-01

    Background An accurate and rapid serologic method to differentiate HIV-2 from HIV-1 infection is required since the confirmatory HIV-1 Western Blot (WB) may demonstrate cross-reactivity with HIV-2 antibodies. Objectives To evaluate the performance of the Bio-Rad Multispot HIV-1/HIV-2 rapid assay as a supplemental test to correctly identify HIV-2 infection and identify HIV-1 WB cross-reactivity with HIV-2 in clinical samples tested at an academic medical center. Study design Between August 2008 and July 2012, clinical samples were screened for HIV using either 3rd-or 4th-generation HIV-1/2 antibody or combination antibody and HIV-1 p24 antigen assays, respectively. All repeatedly reactive samples were reflexed for Multispot rapid testing. Multispot HIV-2 and HIV-1 and HIV-2-reactive samples were further tested using an HIV-2 immunoblot assay and HIV-1 or HIV-2 RNA assays when possible. The HIV-1 WB was performed routinely for additional confirmation and to assess for HIV-2 antibody cross-reactivity. Results Of 46,061 samples screened, 890 (89.6%) of 993 repeatedly reactive samples were also Multispot-reactive: 882 for HIV-1; three for only HIV-2; and five for both HIV-1 and HIV-2. All three HIV-2-only Multispot-positives along with a single dually reactive HIV-1/2 Multispot-positive were also HIV-2 immunoblot-positive; the latter was HIV-1 RNA negative and HIV-2 RNA positive. Conclusions The Multispot rapid test performed well as a supplemental test for HIV-1/2 diagnostic testing. Four new HIV-2 infections (0.45%) were identified from among 890 Multispot-reactive tests. The use of HIV-1 WB alone to confirm HIV-1/2 screening assays may underestimate the true prevalence of HIV-2 infection in the United States. PMID:24342468

  8. Counselor-Based Rapid HIV Testing in Community Pharmacies

    PubMed Central

    Cowan, Ethan; Rhee, John Y.; Brusalis, Christopher; Leider, Jason

    2013-01-01

    Abstract The purpose of this study was to examine the results of implementing a rapid counselor-based HIV testing program in community pharmacies. A prospective cross-sectional study was conducted on a convenience sample of clients at five community pharmacies in New York City (NYC). In 294 days of pharmacy testing, 2805 clients were eligible to receive testing, and 2030 individuals agreed to test. The average age was 33±15 years, 41% were male, 59% were Hispanic, 77% had been previously tested for HIV, and 34% were uninsured. HIV incidence was 0.3%, median CD4 cell count was 622.0, and the average age of the newly diagnosed positives was 36.0±13.9 years. Participants were satisfied with a counselor-based rapid HIV testing program in community-based pharmacies. PMID:23883320

  9. A Rapid, Simple, Effective, and Inexpensive Reconstructed Nipple Flap Guard.

    PubMed

    Khan, Khurrum; Chevray, Pierre M

    2015-10-01

    Nipple reconstruction is a commonly performed component of breast reconstruction. A nipple reconstructed using local skin flaps requires protection from trauma. Here we describe a novel, effective, simple, rapid, inexpensive, and convenient method to protect a reconstructed nipple in the early postoperative period. PMID:26579352

  10. A Rapid, Simple, Effective, and Inexpensive Reconstructed Nipple Flap Guard

    PubMed Central

    Khan, Khurrum

    2015-01-01

    Summary: Nipple reconstruction is a commonly performed component of breast reconstruction. A nipple reconstructed using local skin flaps requires protection from trauma. Here we describe a novel, effective, simple, rapid, inexpensive, and convenient method to protect a reconstructed nipple in the early postoperative period. PMID:26579352

  11. Microfluidic assay without blocking for rapid HIV screening and confirmation.

    PubMed

    Song, Lusheng; Zhang, Yi; Wang, Wenjun; Ma, Liying; Liu, Yong; Hao, Yanlin; Shao, Yiming; Zhang, Wei; Jiang, Xingyu

    2012-08-01

    The essential step for HIV spreading limitation is the screening tests. However, there are multiple disadvantages in current screening assays which need further confirmation test. Herein we developed a rapid HIV assay combining screening and confirmation test by using the microfluidic network assay. Meanwhile, the assay is accelerated by bypassing the step of blocking. We call this method as microfluidic assay without blocking (MAWB). Both the limit of detection and reagent incubation time of MAWB are determined by screening of one model protein pair: ovalbumin and its antibody. The assay time is accelerated about 25% while the limit of detection (LOD) is well kept. Formatting the method in for both HIV screening (testing 8 HIV-related samples) and confirmation (assaying 6 kinds of HIV antibodies of each sample) within 30 min was successful. Fast HIV screening and confirmation of 20 plasma samples were also demonstrated by this method. MAWB improved the assay speed while keeping the LOD of conventional ELISA. Meanwhile, both the accuracy and throughput of MAWB were well improved, which made it an excellent candidate for a quick HIV test for both screening and confirmation. Methods like this one will find wide applications in clinical diagnosis and biochemical analysis based on the interactions between pairs of molecules. PMID:22374476

  12. HIV Rapid Testing in Drug Treatment: Comparison Across Treatment Modalities

    PubMed Central

    Schwartz, Robert P.; Stitzer, Maxine L.; Feaster, Daniel J.; Korthuis, P. Todd; Alvanzo, Anika A. H.; Winhusen, T. M.; Donnard, Lillian; Snead, Ned; Metsch, L. R.

    2012-01-01

    Despite high rates of risky behavior among patients, many drug abuse treatment programs do not provide on-site HIV testing. This secondary analysis examined differences in outcome by program modality from a multi-site trial in which 1,281 HIV-negative patients in 3 methadone programs, 7 non-methadone outpatient programs, and 3 residential programs were randomly assigned to: (1) off-site referral for HIV risk reduction counseling and testing; or on-site rapid testing (2) with or (3) without risk reduction counseling. The parent study using generalized estimating equations with site as a cluster variable found significantly higher rates of HIV testing and feedback of results by 1 month post-enrollment for the combined on-site conditions compared to the offsite condition (RR=4.52, 97.5% CI (3.57, 5.72). Utilizing the same statistical approach, we found neither significant treatment modality nor significant treatment modality by testing condition interaction effects either for receipt of HIV test results at 1 month or for sexual or drug use HIV-risk behaviors at 6-month follow-up. On-site HIV testing is effective across treatment modalities for achieving high rates of testing and results feedback. All programs should be encouraged to adopt or expand this service. PMID:23021496

  13. Simple and rapid determination of myristicin in human serum.

    PubMed

    Dawidowicz, Andrzej L; Dybowski, Michal P

    2013-01-01

    Myristicin (5-allyl-1-methoxy-2,3-methylenodioxybenzene) is the main component of nutmeg (Myristica fragrans Houtt.) essential oil. The increasing use of myristicin as a cheap hallucinogenic intoxicant, frequently causing fatal cases of myristicin poisoning, requires new methods for determination of this compound in blood. This report describes the rapid, simple, and useful procedure for myristicin analysis in human serum, involving myristicin-protein complex degradation before chromatographic analysis. The developed method is characterized by a high recovery (above 99 %), a low detection limit (6.0 ng/g) and good repeatability (average RDS of 2.01 %). PMID:23440626

  14. A simple method for rapidly processing HEU from weapons returns

    SciTech Connect

    McLean, W. II; Miller, P.E.

    1994-01-01

    A method based on the use of a high temperature fluidized bed for rapidly oxidizing, homogenizing and down-blending Highly Enriched Uranium (HEU) from dismantled nuclear weapons is presented. This technology directly addresses many of the most important issues that inhibit progress in international commerce in HEU; viz., transaction verification, materials accountability, transportation and environmental safety. The equipment used to carry out the oxidation and blending is simple, inexpensive and highly portable. Mobile facilities to be used for point-of-sale blending and analysis of the product material are presented along with a phased implementation plan that addresses the conversion of HEU derived from domestic weapons and related waste streams as well as material from possible foreign sources such as South Africa or the former Soviet Union.

  15. Simple and rapid quantification of thrombocytes in zebrafish larvae.

    PubMed

    Huarng, Michael C; Shavit, Jordan A

    2015-06-01

    Platelets are a critical component of hemostasis, with disorders of number or function resulting in coagulation disturbances. Insights into these processes have primarily been realized through studies using mammalian models or tissues. Increasingly, zebrafish embryos and larvae have been used to study the protein and cellular components of hemostasis and thrombosis, including the thrombocyte, a nucleated platelet analog. However, investigations of thrombocytes have been somewhat limited due to lack of a robust and simple methodology for quantitation, an important component of platelet studies in mammals. Using video capture, we have devised an assay that produces a rapid, reproducible, and precise measurement of thrombocyte number in zebrafish larvae by counting fluorescently tagged cells. Averaging 1000 frames, we were able to subtract background fluorescence, thus limiting assessment to circulating thrombocytes. This method facilitated rapid assessment of relative thrombocyte counts in a population of 372 zebrafish larvae by a single operator in less than 3 days. This technique requires basic microscopy equipment and rudimentary programming, lends itself to high throughput analysis, and will enhance future studies of thrombopoiesis in the zebrafish. PMID:25790244

  16. Rapid and simple preparation of a reagentless glucose electrochemical biosensor.

    PubMed

    Zheng, Dan; Vashist, Sandeep Kumar; Al-Rubeaan, Khalid; Luong, John H T; Sheu, Fwu-Shan

    2012-08-21

    A rapid and simple procedure was developed for the preparation of a highly stable and leach-proof glucose oxidase (GOx)-bound glassy carbon electrode (GCE). Crosslinked GOx via glutaraldehyde was drop-cast on a KOH-pretreated GCE followed by drop-casting of 3-aminopropyltriethoxysilane (APTES) to form a stable bioactive layer. At -0.45 V, the biosensor exhibited a wide dynamic detection range of 0.5-48 mM for commercial glucose and 1.3-28.2 mM for Sugar-Chex blood glucose linearity standards. Several endogenous electroactive substances and drug metabolites commonly found in blood were tested and provoked no signal response. To our knowledge, the developed procedure is the most rapid method for preparing a glucose biosensor. The biosensor suffered no biofouling after 7 days of immersion in Sugar-Chex blood glucose. With excellent production reproducibility, GOx-bound electrodes stored dry at room temperature retained their initial activity after several weeks. PMID:22763782

  17. Can Home-Based HIV Rapid Testing Reduce HIV Disparities Among African Americans in Miami?

    PubMed

    Kenya, Sonjia; Okoro, Ikenna S; Wallace, Kiera; Ricciardi, Michael; Carrasquillo, Olveen; Prado, Guillermo

    2016-09-01

    Sixty percent of African Americans have had an HIV test, yet this population disproportionately contributes to AIDS mortality, suggesting that testing is not occurring early enough to achieve optimal outcomes. OraQuick, the first Food and Drug Administration-approved home-based HIV rapid test (HBHRT) could potentially increase testing rates. We assessed whether community health workers (CHWs) paired with HBRHT could improve HIV screening and health care access among African Americans in Miami, Florida. In October-November 2013, 60 African Americans were enrolled and randomized to the experimental condition, which received CHW assistance to complete HBHRT, or the control condition, which were instructed to complete HBHRT independently. Intervention participants were significantly (p ≤ .05) more likely than control participants to complete HBHRT and, if positive, get linked to HIV care (100% vs. 83%) χ(2) (1, N = 60) = 5.46, p ≤ .02. We concluded that CHW-assisted HBHRT may be a promising strategy to improve HIV testing and care among African Americans. PMID:27091604

  18. Rapid HIV Viral Load Suppression in those Initiating Antiretroviral Therapy at First Visit after HIV Diagnosis.

    PubMed

    Hoenigl, Martin; Chaillon, Antoine; Moore, David J; Morris, Sheldon R; Mehta, Sanjay R; Gianella, Sara; Amico, K Rivet; Little, Susan J

    2016-01-01

    Expert guidelines for antiretroviral therapy (ART) now recommend ART as soon as possible in all HIV infected persons to reduce the risk of disease progression and prevent transmission. The goal of this observational study was to evaluate the impact of very early ART initiation and regimen type on time to viral suppression. We evaluated time to viral suppression among 86 persons with newly-diagnosed HIV infection who initiated ART within 30 days of diagnosis. A total of 36 (42%) had acute, 27 (31%) early, and 23 (27%) had established HIV infection. The median time from an offer of immediate ART to starting ART was 8 days. A total of 56/86 (65%) initiated an integrase inhibitor-based regimen and 30/86 (35%) a protease inhibitor-based regimen. The time to viral suppression was significantly shorter in those receiving an integrase inhibitor- versus a protease inhibitor-based regimen (p = 0.022). Twenty-two (26%) initiated ART at their HIV care intake visit and 79% of these participants achieved viral suppression at week 12, 82% at week 24 and 88% at week 48. ART initiated at the intake visit led to rapid and reliable viral suppression in acute, early and chronic HIV infection, in particular when integrase inhibitor-based regimens were used. PMID:27597312

  19. Rapid HIV Viral Load Suppression in those Initiating Antiretroviral Therapy at First Visit after HIV Diagnosis

    PubMed Central

    Hoenigl, Martin; Chaillon, Antoine; Moore, David J.; Morris, Sheldon R.; Mehta, Sanjay R.; Gianella, Sara; Amico, K. Rivet; Little, Susan J.

    2016-01-01

    Expert guidelines for antiretroviral therapy (ART) now recommend ART as soon as possible in all HIV infected persons to reduce the risk of disease progression and prevent transmission. The goal of this observational study was to evaluate the impact of very early ART initiation and regimen type on time to viral suppression. We evaluated time to viral suppression among 86 persons with newly-diagnosed HIV infection who initiated ART within 30 days of diagnosis. A total of 36 (42%) had acute, 27 (31%) early, and 23 (27%) had established HIV infection. The median time from an offer of immediate ART to starting ART was 8 days. A total of 56/86 (65%) initiated an integrase inhibitor-based regimen and 30/86 (35%) a protease inhibitor-based regimen. The time to viral suppression was significantly shorter in those receiving an integrase inhibitor- versus a protease inhibitor-based regimen (p = 0.022). Twenty-two (26%) initiated ART at their HIV care intake visit and 79% of these participants achieved viral suppression at week 12, 82% at week 24 and 88% at week 48. ART initiated at the intake visit led to rapid and reliable viral suppression in acute, early and chronic HIV infection, in particular when integrase inhibitor-based regimens were used. PMID:27597312

  20. Preferences for rapid point-of-care HIV testing in primary care.

    PubMed

    Schwandt, Michael; Nicolle, Eileen; Dunn, Sheila

    2012-01-01

    Although the identification of individuals infected with HIV is an important element of treatment and prevention programs, many people living with HIV are unaware of their status. Thus, individuals are unable to benefit from treatment, and preventable HIV transmission continues to occur. Rapid point-of-care testing for HIV has been found to be preferred by patients in some contexts. However, few studies have examined preferences in primary care populations. This study investigates HIV testing preferences within an urban primary care clinic. Employing a cross-sectional design, data were collected on demographic characteristics, HIV risk factors, and testing history and preferences of participants. A total of 81% of participants stated that they would prefer rapid testing to standard testing, a finding that is consistent across demographic variables and risk factors examined. Increased availability of this modality may decrease barriers to HIV testing, with positive implications both for clinical management of HIV infection and prevention of HIV transmission. PMID:22247336

  1. Using the rapid HIV test to rescreen women in the third trimester of pregnancy.

    PubMed

    Criniti, Shannon M; Aaron, Erika; Levine, Amy B

    2009-01-01

    Antiretroviral therapy during pregnancy in HIV-infected women has dramatically reduced the rate of mother to child HIV transmission in the United States. National guidelines strongly recommend universal HIV testing of all pregnant women with repeat screening in the third-trimester in high-risk populations. To determine patient attitudes towards third-trimester rescreening, a convenience sample was recruited during routine prenatal visits at an urban clinic and participants were surveyed to determine attitudes about HIV third-trimester retesting, acceptability of the rapid HIV testing, condom use, and knowledge of partner's HIV status during pregnancy. Participants were offered a third-trimester rapid HIV retest with the option to decline the test. Eighty pregnant women participated; 95% agreed to be retested with a rapid HIV test, 100% received immediate HIV results, and 91% reported that the rapid test was less stressful than conventional testing. There were no seroconversions. Although 35% did not know their partner's HIV status, 57% of these women reported never using condoms during pregnancy. There was a significant association between reported stage of behavior change and reported likelihood of using condoms. We found that rescreening with the rapid HIV test in the third trimester of pregnancy was well accepted and is important to prevent perinatal HIV transmission. PMID:19879522

  2. Simple isoquinoline and benzylisoquinoline alkaloids as potential antimicrobial, antimalarial, cytotoxic, and anti-HIV agents.

    PubMed

    Iwasa, K; Moriyasu, M; Tachibana, Y; Kim, H S; Wataya, Y; Wiegrebe, W; Bastow, K F; Cosentino, L M; Kozuka, M; Lee, K H

    2001-11-01

    Twenty-six simple isoquinolines and 21 benzylisoquinolines were tested for antimicrobial, antimalarial, cytotoxic, and anti-HIV activities. Some simple isoquinoline alkaloids were significantly active in each assay, and may be useful as lead compounds for developing potential chemotherapeutic agents. These compounds include 13 (antimicrobial), 25, 26, and 42 (antimalarial), 13 and 25 (cytotoxic), and 28 and 29 (anti-HIV). A quaternary nitrogen atom of isoquinolium or dihydroisoquinolinium type may contribute to enhanced potency in the first three types of activities. In contrast, anti-HIV activity was found with tetrahydroisoquinoline and 6,7-dihydroxyisoquinolium salts. PMID:11597468

  3. Rapid and simple method for purification of nucleic acids.

    PubMed Central

    Boom, R; Sol, C J; Salimans, M M; Jansen, C L; Wertheim-van Dillen, P M; van der Noordaa, J

    1990-01-01

    We have developed a simple, rapid, and reliable protocol for the small-scale purification of DNA and RNA from, e.g., human serum and urine. The method is based on the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate together with the nucleic acid-binding properties of silica particles or diatoms in the presence of this agent. By using size-fractionated silica particles, nucleic acids (covalently closed circular, relaxed circular, and linear double-stranded DNA; single-stranded DNA; and rRNA) could be purified from 12 different specimens in less than 1 h and were recovered in the initial reaction vessel. Purified DNA (although significantly sheared) was a good substrate for restriction endonucleases and DNA ligase and was recovered with high yields (usually over 50%) from the picogram to the microgram level. Copurified rRNA was recovered almost undegraded. Substituting size-fractionated silica particles for diatoms (the fossilized cell walls of unicellular algae) allowed for the purification of microgram amounts of genomic DNA, plasmid DNA, and rRNA from cell-rich sources, as exemplified for pathogenic gram-negative bacteria. In this paper, we show representative experiments illustrating some characteristics of the procedure which may have wide application in clinical microbiology. Images PMID:1691208

  4. Rapid progression to gummatous syphilitic hepatitis and neurosyphilis in a patient with newly-diagnosed HIV.

    PubMed

    Pilozzi-Edmonds, Laura; Kong, Ling Yuan; Szabo, Jason; Birnbaum, Leora M

    2015-11-01

    We review the literature on hepatic involvement in patients with HIV and syphilis co-infection and describe a case of rapid progression to neurosyphilis and presumed gummatous syphilitic hepatitis in a patient newly diagnosed with HIV. To our knowledge, this is the first case of syphilitic hepatitis with gummas described in the HIV population. PMID:25525055

  5. Addressing Unmet Need for HIV Testing in Emergency Care Settings: A Role for Computer-facilitated Rapid HIV Testing?

    PubMed Central

    Kurth, Ann E.; Severynen, Anneleen; Spielberg, Freya

    2014-01-01

    HIV testing in emergency departments (EDs) remains underutilized. We evaluated a computer tool to facilitate rapid HIV testing in an urban ED. Randomly assigned non-acute adult ED patients to computer tool (‘CARE’) and rapid HIV testing before standard visit (n=258) or to standard visit (n=259) with chart access. Assessed intervention acceptability and compared noted HIV risks. Participants were 56% non-white, 58% male; median age 37 years. In the CARE arm nearly all (251/258) completed the session and received HIV results; 4 declined test consent. HIV risks were reported by 54% of users and there was one confirmed HIV-positive and 2 false-positives (seroprevalence 0.4%, 95% CI 0.01–2.2%). Half (55%) preferred computerized, over face-to-face, counseling for future HIV testing. In standard arm, one HIV test and 2 referrals for testing occurred. Computer-facilitated HIV testing appears acceptable to ED patients. Future research should assess cost-effectiveness compared with staff-delivered approaches. PMID:23837807

  6. Rapid HIV testing for developing countries: the challenge of false-negative tests

    NASA Astrophysics Data System (ADS)

    Yogev, Ram

    2012-06-01

    It is a common practice in resource-constrained countries to accept two positive rapid HIV antibody test results as diagnostic for HIV infection. Because these tests are inexpensive and results are obtained quickly, they are recommended by the WHO to "scale-up" HIV testing to increase the number of people tested. The negative predictive value of rapid HIV tests is so high that negative results are considered conclusive despite the fact that false-negative results can occur in several situations. While the specificity and sensitivity of rapid HIV tests in resource-rich countries is acceptable, there are only limited data about their performance in resource-constrained countries. The challenges of rapid HIV testing in these situations will be discussed.

  7. Rapid HIV Testing for Individuals on Probation/Parole: Outcomes of an Intervention Trial

    PubMed Central

    Gordon, Michael S.; Kinlock, Timothy W.; McKenzie, Michelle; Wilson, Monique E.; Rich, Josiah D.

    2013-01-01

    Many probationers and parolees do not receive HIV testing despite being at increased risk for obtaining and transmitting HIV. A two-group randomized controlled trial was conducted between April, 2011 and May, 2012 at probation/parole offices in Baltimore, Maryland and Providence/Pawtucket, Rhode Island. Male and female probationers/parolees were interviewed (N=1263) and then offered HIV testing based on random assignment to one of two conditions: 1) On-site rapid HIV testing conducted at the probation/parole office; or 2) Referral for rapid HIV testing off site at a community HIV testing clinic. Outcomes were: 1) undergoing HIV testing; and 2) receipt of HIV testing results. Participants were significantly more likely to be tested onsite at a probation/parole office versus off-site at a HIV testing clinic (p < .001). There was no difference between the two groups in terms of receiving HIV testing results. Findings indicate that probationers/ parolees are willing to be tested on-site and, independent of testing location, are equally willing to receive their results. Implications for expanding rapid HIV testing to more criminal justice related locations and populations are discussed. PMID:23536140

  8. Rapid testing at labor and delivery to prevent mother-to-child HIV transmission in developing settings: issues and challenges.

    PubMed

    Pai, Nitika Pant; Klein, Marina B

    2009-01-01

    Worldwide, approximately 2.5 million children (95% CI: 2.2-2.6) are living with HIV infection. In 2007 alone, approximately 420,000 children (95%CI: 350,000-540,000) were newly infected with HIV - a vast majority of these infections were acquired through maternal-fetal transmission. Many of these infections could have been reduced by timely diagnosis and the delivery of interventions aimed at preventing mother-to-child HIV transmission. This perspective examines the attitudes preventing women from accessing HIV testing early on during pregnancy and the issues and challenges that remain in the institutionalization of interventions to prevent mother-to-child HIV transmission at labor and delivery. Socio-cultural and economic factors prevent women from accessing testing at an opportune time during pregnancy. In addition, a lack of adequate infrastructure often prevents timely delivery of interventions to those who access testing at the last minute (i.e., during labor and delivery). In the wake of a pediatric HIV epidemic and the need for lifelong provision of antiretroviral therapy to infected children, a simple strategy for provision of round-the-clock rapid testing and counseling services in the labor rooms may be cost saving to the healthcare systems worldwide. PMID:19102641

  9. Laboratory Evaluation of Three Rapid Diagnostic Tests for Dual Detection of HIV and Treponema pallidum Antibodies

    PubMed Central

    Woo, Jennifer S.; Chung, Jun Ho; Sokovic, Anita; Bristow, Claire C.; Klausner, Jeffrey D.

    2014-01-01

    The performance of three research-use-only, dual HIV and syphilis rapid diagnostic tests (RDTs) was evaluated for 150 patient serum samples and compared to reference HIV and Treponema pallidum antibody detection methods. The RDTs performed comparably, with sensitivities of 93 to 99% and specificities of 97 to 100%. The kappa statistic between the RDTs was 0.95. PMID:25297332

  10. Laboratory evaluation of three rapid diagnostic tests for dual detection of HIV and Treponema pallidum antibodies.

    PubMed

    Humphries, Romney M; Woo, Jennifer S; Chung, Jun Ho; Sokovic, Anita; Bristow, Claire C; Klausner, Jeffrey D

    2014-12-01

    The performance of three research-use-only, dual HIV and syphilis rapid diagnostic tests (RDTs) was evaluated for 150 patient serum samples and compared to reference HIV and Treponema pallidum antibody detection methods. The RDTs performed comparably, with sensitivities of 93 to 99% and specificities of 97 to 100%. The kappa statistic between the RDTs was 0.95. PMID:25297332

  11. Rapid evolution of simple sequence repeat induced by allopolyploidization.

    PubMed

    Tang, Zongxiang; Fu, Shulan; Ren, Zhenglong; Zou, Yuting

    2009-09-01

    Microsatellite evolution normally occurs in diploids. Until now, there has been a lack of direct experimental evidence for microsatellite evolution following allopolyploidization. In the present study, F(1) hybrids and newly synthesized allopolyploids were derived from Triticum aestivum Chinese Spring x Secale cereale Jinzhou-heimai. One hundred and sixty-three wheat simple sequence repeat (SSR) markers were used to investigate the variation of wheat microsatellites after allopolyploidization and variation of the PCR products of 29 of the SSR markers was observed. Of these 29 SSR markers, 15 were unable to produce products from amphiploids. The other 14 SSR markers did produce products from parental wheat, F(1) hybrids and amphiploids. However, the length of the products amplified from amphiploids was different from the length of the products amplified from parental wheat and F(1) hybrids. Sequencing indicated that the length variation of the 14 microsatellites stemmed mainly from variation in the number of repeat units. The alteration of repeat units occurred in both perfect and compound repeats. In some compound SSR loci, one motif was observed to expand whereas another to contract. Almost all the microsatellite evolution observed in this study could be explained by the slipped-strand mispairing model. The results of this study seem to indicate that stress caused by allopolyploidization might be one of the factors that induce microsatellite evolution. In addition, the findings of present study provided an instance of how simple sequence repeats evolved after allopolyploidization. PMID:19688286

  12. Simple, rapid method for the preparation of isotopically labeled formaldehyde

    DOEpatents

    Hooker, Jacob Matthew; Schonberger, Matthias; Schieferstein, Hanno; Fowler, Joanna S.

    2011-10-04

    Isotopically labeled formaldehyde (*C.sup..sctn.H.sub.2O) is prepared from labeled methyl iodide (*C.sup..sctn.H.sub.3I) by reaction with an oxygen nucleophile having a pendant leaving group. The mild and efficient reaction conditions result in good yields of *C.sup..sctn.H.sub.2O with little or no *C isotopic dilution. The simple, efficient production of .sup.11CH.sub.2O is described. The use of the .sup.11CH.sub.2O for the formation of positron emission tomography tracer compounds is described. The reaction can be incorporated into automated equipment available to radiochemistry laboratories. The isotopically labeled formaldehyde can be used in a variety of reactions to provide radiotracer compounds for imaging studies as well as for scintillation counting and autoradiography.

  13. A Rapid and Simple Bioassay Method for Herbicide Detection

    PubMed Central

    Li, Xiu-Qing; Ng, Alan; King, Russell; Durnford, Dion G.

    2008-01-01

    Chlamydomonas reinhardtii, a unicellular green alga, has been used in bioassay detection of a variety of toxic compounds such as pesticides and toxic metals, but mainly using liquid culture systems. In this study, an algal lawn—agar system for semi-quantitative bioassay of herbicidal activities has been developed. Sixteen different herbicides belonging to 11 different categories were applied to paper disks and placed on green alga lawns in Petri dishes. Presence of herbicide activities was indicated by clearing zones around the paper disks on the lawn 2–3 days after application. The different groups of herbicides induced clearing zones of variable size that depended on the amount, mode of action, and chemical properties of the herbicides applied to the paper disks. This simple, paper-disk-algal system may be used to detect the presence of herbicides in water samples and act as a quick and inexpensive semi-quantitative screening for assessing herbicide contamination. PMID:19578512

  14. Rapid Immuno-Chromatographic Assay for the Detection of Antibodies to HIV Compare with Elisa among Voluntary and Replacement Blood Donor of Mymensingh Medical College Hospital.

    PubMed

    Chakrabarty, P; Rudra, S; Hossain, M A; Begum, S A; Mirza, T T; Rudra, M

    2015-04-01

    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 voluntary and replacement blood donors & HIV-infected patients (positive samples from BSMMU, Dhaka). 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 (Biotech) were evaluated between 1st February to 30th June, 2013 using 400 whole blood samples from voluntary and replacement 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). Only 01 sample including ten positive samples from BSMMU were confirmed HIV-1 antibody positive, while 399 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 concordantly false positive in Uni-Gold™, Determine and SD Bioline assays. An alternative confirmatory HIV testing strategy based on initial testing on either SD Bioline or Determine assays followed by testing of reactive

  15. Effect of rapid HIV testing on HIV incidence and services in populations at high risk for HIV exposure: an equity-focused systematic review

    PubMed Central

    Pottie, Kevin; Medu, Olanrewaju; Welch, Vivian; Dahal, Govinda P; Tyndall, Mark; Rader, Tamara; Wells, George

    2014-01-01

    Objective To assess the effects of rapid voluntary counselling and testing (VCT) for HIV on HIV incidence and uptake of HIV/AIDS services in people at high risk for HIV exposure. Design Cochrane systematic review and meta-analysis. Data sources We searched PubMed, EMBASE, AIDSearch, LILACS, Global Health, Medline Africa, PsychInfo, CINAHL, Cochrane CENTRAL, Cochrane HIV/AIDS Group Specialized Register and grey literature from 1 January 2001 to 5 June 2014 without language restriction. Data selection We included controlled studies that compared rapid VCT with conventional testing among people at risk for HIV exposure. Data extraction Two reviewers extracted data. We used Cochrane risk of bias tool and GRADE criteria: risk of bias, inconsistency, indirectness, imprecision and publication bias. For observational studies we used the Newcastle-Ottawa Scale. We used the PRISMA-Equity reporting guideline. Results From 2441 articles, we included 8 randomised controlled trials and 5 observational studies. Rapid VCT was associated with a threefold increase in HIV-testing uptake (relative risk (RR)=2.95 95% CI 1.69 to 5.16) and a twofold increase in the receipt of test results (RR=2.14, 95% CI 1.08 to 4.24). Women accepted testing more often than men in rapid VCT arm, but no differences in effect for age or socioeconomic status. Observational studies also showed rapid VCT led to higher rates of uptake of testing. Heterogeneity was high. A cluster-randomised trial reported an 11% reduction in HIV incidence in intervention communities (RR=0.89, 95% CI=0.63 to 1.24) over 3 years trial. Conclusions Rapid VCT in health facilities and communities was associated with a large increase in HIV-testing uptake and receipt of results. This has implications for WHO guidelines. The routine use of rapid VCT may also help avoid human rights violations among marginalised populations where testing may occur without informed consent and where existing stigma may create barriers to testing

  16. Rapid HIV Testing on the College Campus: Comparing Traditional and Outreach Models.

    PubMed

    Przybyla, Sarahmona M

    2013-01-01

    The purpose of this study was to compare rapid HIV testing services on a college campus between a clinic-based testing group and an outreach-based testing group. Study participants were 1,233 individuals who underwent HIV counseling and testing. Questionnaires assessed demographics and HIV transmission risk behaviors. Results indicate that outreach-based testers were more likely to be younger, female, and African American relative to clinic-based testers. Overall, 100% of clinic-based testers and 99.5% of outreach-based testers receiving their test results. All individuals with positive rapid test results received confirmatory blood testing and entered medical care within one week of preliminary diagnosis. College campuses may provide a unique setting to deliver HIV testing and may help increase the percentage of young people who are aware of their serostatus, particularly younger, female, and African American students who may be less likely to undergo testing in traditional clinic settings. PMID:24416620

  17. Offer of rapid testing and alternative biological samples as practical tools to implement HIV screening programs.

    PubMed

    Parisi, Maria Rita; Soldini, Laura; Di Perri, Giovanni; Tiberi, Simon; Lazzarin, Adriano; Lillo, Flavia B

    2009-10-01

    Implementation of HIV testing has the objective to increase screening, identify and counsel persons with infection, link them to clinical services and reduce transmission. Rapid tests and/or alternative biological samples (like oral fluid) give the option for a better general consent in approaching screening, immediate referral of HIV positives to medical treatment and partner notification. We tested the performance characteristics of an oral fluid-based rapid HIV test (Rapidtest HIV lateral flow-Healthchem diag. LLC) in comparison with routinely utilized methods in a selected population of known positive (N = 121) or negative (N = 754) subjects. The sensitivity of the rapid test was 99.1% (one false negative sample) and the specificity 98.8%. Five negatives showed a faint reactivity, 3 of these were reactive also in the reference test, one with a p24 only reaction in Western blot. If these 3 samples were excluded from the analysis the specificity increases to 99.2%. Results from our study confirm that, although a continuous improvement of the test performance is still needed to minimize false negative and positive results, rapid test and alternative biological samples may contribute to HIV prevention strategies by reaching a larger population particularly when and where regular screening procedures are difficult to obtain. PMID:20128446

  18. HIV rapid testing in a Veterans Affairs hospital ED setting: a 5-year sustainability evaluation.

    PubMed

    Knapp, Herschel; Hagedorn, Hildi; Anaya, Henry D

    2014-08-01

    Routine HIV testing in primary care settings is now recommended in the United States. The US Department of Veterans Affairs (VA) has increased the number of patients tested for HIV, but overall HIV testing rates in VA remain low. A proven strategy for increasing such testing involves nurse-initiated HIV rapid testing (HIV RT). The purpose of this work was to use a mixed methodology approach to evaluate the 5-year sustainability of an intervention that implemented HIV RT in a VA emergency department setting in a large, urban VA medical center to reduce missed diagnostic and treatment opportunities in this vulnerable patient population. In-person semistructured interviews were conducted with providers and stakeholders. Interview notes were qualitatively coded for emerging themes. Quarterly testing rates were evaluated for a 5-year time span starting from the launch in July 2008. Findings indicate that HIV RT was sustained by the enthusiasm of 2 clinical champions who oversaw the registered nurses responsible for conducting the testing. The departure of the clinical champions was correlated with a substantial drop-off in testing. Findings also indicate potential strategies for improving sustainability including engaging senior leadership in the project, engaging line staff in the implementation planning from the start to increase ownership over the innovation, incorporating information into initial training explaining the importance of the innovation to quality patient care, providing ongoing training to maintain skills, and providing routine progress reports to staff to demonstrate the ongoing impact of their efforts. PMID:24908442

  19. Triggering HIV polyprotein processing by light using rapid photodegradation of a tight-binding protease inhibitor

    PubMed Central

    Schimer, Jiří; Pávová, Marcela; Anders, Maria; Pachl, Petr; Šácha, Pavel; Cígler, Petr; Weber, Jan; Majer, Pavel; Řezáčová, Pavlína; Kräusslich, Hans-Georg; Müller, Barbara; Konvalinka, Jan

    2015-01-01

    HIV protease (PR) is required for proteolytic maturation in the late phase of HIV replication and represents a prime therapeutic target. The regulation and kinetics of viral polyprotein processing and maturation are currently not understood in detail. Here we design, synthesize, validate and apply a potent, photodegradable HIV PR inhibitor to achieve synchronized induction of proteolysis. The compound exhibits subnanomolar inhibition in vitro. Its photolabile moiety is released on light irradiation, reducing the inhibitory potential by 4 orders of magnitude. We determine the structure of the PR-inhibitor complex, analyze its photolytic products, and show that the enzymatic activity of inhibited PR can be fully restored on inhibitor photolysis. We also demonstrate that proteolysis of immature HIV particles produced in the presence of the inhibitor can be rapidly triggered by light enabling thus to analyze the timing, regulation and spatial requirements of viral processing in real time. PMID:25751579

  20. Triggering HIV polyprotein processing by light using rapid photodegradation of a tight-binding protease inhibitor.

    PubMed

    Schimer, Jiří; Pávová, Marcela; Anders, Maria; Pachl, Petr; Šácha, Pavel; Cígler, Petr; Weber, Jan; Majer, Pavel; Řezáčová, Pavlína; Kräusslich, Hans-Georg; Müller, Barbara; Konvalinka, Jan

    2015-01-01

    HIV protease (PR) is required for proteolytic maturation in the late phase of HIV replication and represents a prime therapeutic target. The regulation and kinetics of viral polyprotein processing and maturation are currently not understood in detail. Here we design, synthesize, validate and apply a potent, photodegradable HIV PR inhibitor to achieve synchronized induction of proteolysis. The compound exhibits subnanomolar inhibition in vitro. Its photolabile moiety is released on light irradiation, reducing the inhibitory potential by 4 orders of magnitude. We determine the structure of the PR-inhibitor complex, analyze its photolytic products, and show that the enzymatic activity of inhibited PR can be fully restored on inhibitor photolysis. We also demonstrate that proteolysis of immature HIV particles produced in the presence of the inhibitor can be rapidly triggered by light enabling thus to analyze the timing, regulation and spatial requirements of viral processing in real time. PMID:25751579

  1. Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method.

    PubMed

    McFall, Sally M; Neto, Mário F; Reed, Jennifer L; Wagner, Robin L

    2016-01-01

    FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad to extract cellular DNA from whole blood in less than 2 min. The blood specimen is treated with detergent, mixed briefly and applied by pipet to the separation membrane. The lysate wicks into the blotting pad due to capillary action, capturing the genomic DNA on the surface of the separation membrane. The extracted DNA is retained on the membrane during a simple wash step wherein PCR inhibitors are wicked into the absorbent blotting pad. The membrane containing the entrapped DNA is then added to the PCR reaction without further purification. This simple method does not require laboratory equipment and can be easily implemented with inexpensive laboratory supplies. Here we describe a protocol for highly sensitive detection and quantitation of HIV-1 proviral DNA from 100 µl whole blood as a model for early infant diagnosis of HIV that could readily be adapted to other genetic targets. PMID:27583575

  2. A simple, rapid and inexpensive screening method for the identification of Pythium insidiosum.

    PubMed

    Tondolo, Juliana Simoni Moraes; Loreto, Erico Silva; Denardi, Laura Bedin; Mario, Débora Alves Nunes; Alves, Sydney Hartz; Santurio, Janio Morais

    2013-04-01

    Growth of Pythium insidiosum mycelia around minocycline disks (30μg) did not occur within 7days of incubation at 35°C when the isolates were grown on Sabouraud, corn meal, Muller-Hinton or RPMI agar. This technique offers a simple and rapid method for the differentiation of P. insidiosum from true filamentous fungi. PMID:23419825

  3. Coins and Costs: A Simple and Rapid Assessment of Basic Financial Knowledge

    ERIC Educational Resources Information Center

    Willner, Paul; Bailey, Rebecca; Dymond, Simon; Parry, Rhonwen

    2011-01-01

    Introduction: We describe a simple and rapid screening test for basic financial knowledge that is suitable for administration to people with mild intellectual disabilities. Method: The Coins and Costs test asks respondents to name coins, and to estimate prices of objects ranging between 1 British Pound (an ice cream) and 100K British Pounds (a…

  4. Setaria viridis floral-dip: A simple and rapid Agrobacterium-medicated transformation method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plan...

  5. A simple and rapid method for the analysis of phenolic compounds in beverages and grains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A simple, rapid and direct method for detection of phenolics in foods and beverages is needed. The current standard method (Folin-Ciocalteau) indirectly measures the “total phenolics” through the reducing capacity of components of food or beverages. A novel method was developed to quantify polypheno...

  6. Rapidly Cleared Episodes of Oral and Anogenital Herpes Simplex Virus Shedding in HIV-Infected Adults

    PubMed Central

    Mark, Karen E.; Wald, Anna; Magaret, Amalia S.; Selke, Stacy; Kuntz, Steven; Huang, Meei-Li; Corey, Lawrence

    2011-01-01

    Objective To determine whether rapidly cleared episodes of herpes simplex virus (HSV) reactivation occur in HIV-infected adults. Methods Twenty HSV-2 seropositive, HIV seropositive adults, including 9 (45%) who were also HSV-1 seropositive, collected oral and anogenital swabs for HSV DNA PCR 4 times a day for 60 days. Samples were positive for HSV if we detected ≥ 150 copies of HSV DNA/mL of specimen. Results Median HSV shedding episode duration was 7.5 (range 4–253) hours for oral and 11 (range 4–328) hours for anogenital reactivation. Thirty-five percent of oral and 29% of anogenital reactivations lasted ≤ 6 hours, and 59% of oral and 53% of anogenital reactivations lasted ≤ 12 hours. Seven of 9 participants who shed orally and 10 of 15 who shed anogenitally had ≥ 1 reactivation lasting ≤ 6 hours. The median maximum level of HSV DNA detected in an episode increased with episode duration for both oral and anogenital episodes. Concurrent oral and anogenital shedding occurred more frequently than expected: Oral HSV shedding was detected on 17% of time points with anogenital, but 1% of time points without anogenital, shedding (p < 0.001). Conclusions Rapidly cleared episodes of oral and anogenital HSV shedding occur in HIV-infected persons, supporting the hypothesis that frequent anogenital mucosal immune activation caused by HSV-2 is present in HIV co-infected persons, potentially contributing to HIV infectiousness. PMID:20616743

  7. Isolation of human immunodeficiency virus type 1 (HIV-1) RNA from feces by a simple method and difference between HIV-1 subpopulations in feces and serum.

    PubMed Central

    van der Hoek, L; Boom, R; Goudsmit, J; Snijders, F; Sol, C J

    1995-01-01

    A simple method for the isolation and subsequent detection of human immunodeficiency virus type 1 (HIV-1) RNA from feces is described. Viral RNA was isolated by the method developed by Boom et al. (R. Boom, C.J.A. Sol, M.M.M. Salimans, C.L. Jansen, P.M.E. Wertheim-van Dillen, and J. van der Noordaa, J. Clin. Microbiol. 28:495-503, 1990), which was adapted for feces. HIV-1 RNA was detected by reverse transcription (RT) followed by a nested PCR encompassing the V3 region. Reconstruction experiments revealed that the efficiencies of the extraction technique and the subsequent RT-PCR were not considerably affected by the varied composition of feces. The method was applied on fecal specimens from 18 HIV-1-infected individuals, among which were samples that had been stored for 9 years. It appeared that HIV-1 RNA was detectable in the feces of 12 persons (67%). Viral RNA was present in the feces of persons who fulfilled the criteria for CDC class II and CDC class III HIV infection as well as in patients who were diagnosed with AIDS (CDC class IV). Direct sequencing of amplimers obtained from paired fecal and serum specimens showed that differences in sequence heterogeneity existed. In one patient a remarkable difference in the HIV-1 sequences between isolates from feces and serum was observed. In conclusion, HIV-1 RNA is frequently present in the feces of HIV-1-infected individuals, and in some cases the HIV-1 subpopulation in feces differs from the HIV-1 subpopulation in serum. PMID:7751361

  8. Rapid HIV Testing and Counseling for Residents in Domestic Violence Shelters

    PubMed Central

    Draucker, Claire Burke; Johnson, Dawn M.; Johnson, Nicole L.; Kadeba, Myriam T.; Mazurczyk, Jill; Zlotnick, Caron

    2015-01-01

    Over one million Americans live with the human immunodeficiency virus (HIV), and roughly 20% of those living with HIV are unaware of their status. One way to decrease this epidemic is community-based rapid testing with high-risk populations. One high-risk population that has received limited attention is victims of intimate partner violence (IPV) who seek shelter. In an effort to gain foundational information to implement rapid HIV testing and counseling services in domestic violence shelters, the current study conducted a series of focus groups with 18 residents and 10 staff of local shelters from October 15th to December 12th, 2012. Participants provided valuable insight into how HIV rapid testing and counseling might be best implemented given the resources and constraints of shelter life. Despite identifying some potential barriers, most believed that the promise of quick results, the convenience and support afforded by the shelter venue, and the timing of the intervention at a point when women are making life changes would render the intervention acceptable to residents. Further insights are discussed in the article. PMID:25738795

  9. A simple bridging flocculation assay for rapid, sensitive and stringent detection of gene specific DNA methylation.

    PubMed

    Wee, Eugene J H; Ha Ngo, Thu; Trau, Matt

    2015-01-01

    The challenge of bringing DNA methylation biomarkers into clinic is the lack of simple methodologies as most current assays have been developed for research purposes. To address the limitations of current methods, we describe herein a novel methyl-protein domain (MBD) enrichment protocol for simple yet rapid and highly stringent selection of highly methylated DNA from limiting input samples. We then coupled this with a DNA-mediated flocculation assay for rapid and low cost naked-eye binary evaluation of highly methylated genes in cell line and blood DNA. The low resource requirements of our method may enable widespread adoption of DNA methylation-based diagnostics in clinic and may be useful for small-scale research. PMID:26458746

  10. A simple bridging flocculation assay for rapid, sensitive and stringent detection of gene specific DNA methylation

    PubMed Central

    Wee, Eugene J. H.; Ha Ngo, Thu; Trau, Matt

    2015-01-01

    The challenge of bringing DNA methylation biomarkers into clinic is the lack of simple methodologies as most current assays have been developed for research purposes. To address the limitations of current methods, we describe herein a novel methyl-protein domain (MBD) enrichment protocol for simple yet rapid and highly stringent selection of highly methylated DNA from limiting input samples. We then coupled this with a DNA-mediated flocculation assay for rapid and low cost naked-eye binary evaluation of highly methylated genes in cell line and blood DNA. The low resource requirements of our method may enable widespread adoption of DNA methylation-based diagnostics in clinic and may be useful for small-scale research. PMID:26458746

  11. [Benefits of using rapid HIV testing at the PMU-FLON walk-in clinic in Lausanne].

    PubMed

    Gilgien, W; Aubert, J; Bischoff, T; Herzig, L; Perdrix, J

    2012-05-16

    Lab tests are frequently used in primary care to guide patient care. This is particularly the case when a severe disorder, or one that will affect patients' initial care, needs to be excluded rapidly. At the PMU-FLON walk-in clinic the use of HIV testing as recommended by the Swiss Office of Public Health was hampered by the delay in obtaining test results. This led us to introduce rapid HIV testing which provides results within 30 minutes. Following the first 250 tests the authors discuss the results as well as the benefits of rapid HIV testing in an urban walk-in clinic. PMID:22730643

  12. Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages.

    PubMed

    Graziano, Francesca; Desdouits, Marion; Garzetti, Livia; Podini, Paola; Alfano, Massimo; Rubartelli, Anna; Furlan, Roberto; Benaroch, Philippe; Poli, Guido

    2015-06-23

    HIV type 1 (HIV-1) infects CD4(+) T lymphocytes and tissue macrophages. Infected macrophages differ from T cells in terms of decreased to absent cytopathicity and for active accumulation of new progeny HIV-1 virions in virus-containing compartments (VCC). For these reasons, infected macrophages are believed to act as "Trojan horses" carrying infectious particles to be released on cell necrosis or functional stimulation. Here we explored the hypothesis that extracellular ATP (eATP) could represent a microenvironmental signal potentially affecting virion release from VCC of infected macrophages. Indeed, eATP triggered the rapid release of infectious HIV-1 from primary human monocyte-derived macrophages (MDM) acutely infected with the CCR5-dependent HIV-1 strain. A similar phenomenon was observed in chronically infected promonocytic U1 cells differentiated to macrophage-like cells (D-U1) by costimulation with phorbol esters and urokinase-type plasminogen activator. Worthy of note, eATP did not cause necrotic, apoptotic, or pyroptotic cell death, and its effect on HIV-1 release was suppressed by Imipramine (an antidepressant agent known to inhibit microvesicle formation by interfering with membrane-associated acid sphingomyelinase). Virion release was not triggered by oxidized ATP, whereas the effect of eATP was inhibited by a specific inhibitor of the P2X7 receptor (P2X7R). Thus, eATP triggered the discharge of virions actively accumulating in VCC of infected macrophages via interaction with the P2X7R in the absence of significant cytopathicity. These findings suggest that the microvesicle pathway and P2X7R could represent exploitable targets for interfering with the VCC-associated reservoir of infectious HIV-1 virions in tissue macrophages. PMID:26056317

  13. Rapid and simple spectrophotometric determination of persulfate in water by microwave assisted decolorization of Methylene Blue.

    PubMed

    Zhao, Lajuan; Yang, Shiying; Wang, Leilei; Shi, Chao; Huo, Meiqing; Li, Yan

    2015-05-01

    A rapid and simple method for determination of persulfate in aqueous solution was developed. The method is based on the rapid reaction of persulfate with Methylene Blue (MB) via domestic microwave activation, which can promote the activation of persulfate and decolorize MB quickly. The depletion of MB at 644 nm (the maximum absorption wavelength of MB) is in proportion to the increasing concentration of persulfate in aqueous solution. Linear calibration curve was obtained in the range 0-1.5 mmol/L, with a limit of detection of 0.0028 mmol/L. The reaction time is rapid (within 60 sec), which is much shorter than that used for conventional methods. Compared with existing analytical methods, it need not any additives, especially colorful Fe2+, and need not any pretreatment for samples, such as pH adjustment. PMID:25968279

  14. HIV testing of children is not simple for health providers and researchers: Legal and policy frameworks guidance in South Africa.

    PubMed

    Van Rooyen, Heidi Eve; Strode, Ann E; Slack, Catherine M

    2016-01-01

    Antiretroviral treatment coverage for children and adolescents is significantly lower than that for adults. A first step in improving this situation is ensuring increased access to HIV counselling and testing services. Current legal and policy frameworks outline four norms that should inform HIV testing of children in South Africa: limiting HIV testing to defined circumstances, and ensuring that consent is obtained, counselling is provided and confidentiality is maintained. Implementing these norms is not simple. We discuss the challenges and opportunities these norms present for children, their families, health providers and researchers working in this area. Better alignment between evolving public health approaches and the HIV counselling and testing legal/policy frameworks (and the internal coherence of domestic frameworks) would better serve children, their parents and those who work with them. PMID:27138658

  15. Implementing Rapid HIV Testing With or Without Risk-Reduction Counseling in Drug Treatment Centers: Results of a Randomized Trial

    PubMed Central

    Feaster, Daniel J.; Gooden, Lauren; Matheson, Tim; Mandler, Raul N.; Haynes, Louise; Tross, Susan; Kyle, Tiffany; Gallup, Dianne; Kosinski, Andrzej S.; Douaihy, Antoine; Schackman, Bruce R.; Das, Moupali; Lindblad, Robert; Erickson, Sarah; Korthuis, P. Todd; Martino, Steve; Sorensen, James L.; Szapocznik, José; Walensky, Rochelle; Branson, Bernard; Colfax, Grant N.

    2012-01-01

    Objectives. We examined the effectiveness of risk reduction counseling and the role of on-site HIV testing in drug treatment. Methods. Between January and May 2009, we randomized 1281 HIV-negative (or status unknown) adults who reported no past-year HIV testing to (1) referral for off-site HIV testing, (2) HIV risk-reduction counseling with on-site rapid HIV testing, or (3) verbal information about testing only with on-site rapid HIV testing. Results. We defined 2 primary self-reported outcomes a priori: receipt of HIV test results and unprotected anal or vaginal intercourse episodes at 6-month follow-up. The combined on-site rapid testing participants received more HIV test results than off-site testing referral participants (P < .001; Mantel-Haenszel risk ratio = 4.52; 97.5% confidence interval [CI] = 3.57, 5.72). At 6 months, there were no significant differences in unprotected intercourse episodes between the combined on-site testing arms and the referral arm (P = .39; incidence rate ratio [IRR] = 1.04; 97.5% CI = 0.95, 1.14) or the 2 on-site testing arms (P = .81; IRR = 1.03; 97.5% CI = 0.84, 1.26). Conclusions. This study demonstrated on-site rapid HIV testing’s value in drug treatment centers and found no additional benefit from HIV sexual risk-reduction counseling. PMID:22515871

  16. Rapid and sensitive detection of HIV-1 p24 antigen by immunomagnetic separation coupled with catalytic fluorescent immunoassay.

    PubMed

    Zhang, Yun; Yang, Hang; Yu, Junping; Wei, Hongping

    2016-09-01

    In this study, a system of magnetic beads (MBs) coupled with catalytic fluorescent immunoassay for rapid and sensitive determination of HIV-1 capsid antigen p24 was developed. p24 was captured by antibody immobilized MBs, and the detection antibody was linked to horseradish peroxidase (HRP) through biotin-streptavidin recognition, catalyzing the oxidation of o-phenylenediamine (OPD) and hydrogen peroxide to produce a fluorescent product. This is the first reported utilization of the fluorescence of OPD oxidation product catalyzed by HRP for immunoassay. Optimization of conditions afforded a low detection limit of 0.5 pg/mL (3σ) for p24 with a linear range of 1.4-90.0 pg/mL. The assay exhibited good reproducibility with a relative standard deviation (RSD) of 4.4 %, 4.7 %, and 5.0 % for detecting 1.4 pg/mL, 22.5 pg/mL, and 45.0 pg/mL p24, respectively. The assay can be completed in less than 90 min. Moreover, the proposed method was successfully applied to detect p24 in spiked serum. This method overcomes the interference of MBs to the fluorescence signal and demonstrated higher sensitivity for detection of p24 than conventional ELISA kits. The system could be applied for detecting other antigens with high sensitivity, rapidity, specificity, and simple operation. Graphical Abstract A rapid and sensitive biosensing method coupling immunomagnetic separation and catalytic fluorescence for determination of HIV-1 p24 has been developed. PMID:27351993

  17. Compression-based distance (CBD): a simple, rapid, and accurate method for microbiota composition comparison

    PubMed Central

    2013-01-01

    Background Perturbations in intestinal microbiota composition have been associated with a variety of gastrointestinal tract-related diseases. The alleviation of symptoms has been achieved using treatments that alter the gastrointestinal tract microbiota toward that of healthy individuals. Identifying differences in microbiota composition through the use of 16S rRNA gene hypervariable tag sequencing has profound health implications. Current computational methods for comparing microbial communities are usually based on multiple alignments and phylogenetic inference, making them time consuming and requiring exceptional expertise and computational resources. As sequencing data rapidly grows in size, simpler analysis methods are needed to meet the growing computational burdens of microbiota comparisons. Thus, we have developed a simple, rapid, and accurate method, independent of multiple alignments and phylogenetic inference, to support microbiota comparisons. Results We create a metric, called compression-based distance (CBD) for quantifying the degree of similarity between microbial communities. CBD uses the repetitive nature of hypervariable tag datasets and well-established compression algorithms to approximate the total information shared between two datasets. Three published microbiota datasets were used as test cases for CBD as an applicable tool. Our study revealed that CBD recaptured 100% of the statistically significant conclusions reported in the previous studies, while achieving a decrease in computational time required when compared to similar tools without expert user intervention. Conclusion CBD provides a simple, rapid, and accurate method for assessing distances between gastrointestinal tract microbiota 16S hypervariable tag datasets. PMID:23617892

  18. Operational feasibility of using whole blood in the rapid HIV testing algorithm of a resource-limited settings like Bangladesh

    PubMed Central

    Munshi, Saif U.; Oyewale, Tajudeen O.; Begum, Shahnaz; Uddin, Ziya; Tabassum, Shahina

    2016-01-01

    Background Serum-based rapid HIV testing algorithm in Bangladesh constitutes operational challenge to scaleup HIV testing and counselling (HTC) in the country. This study explored the operational feasibility of using whole blood as alternative to serum for rapid HIV testing in Bangladesh. Methods Whole blood specimens were collected from two study groups. The groups included HIV-positive patients (n = 200) and HIV-negative individuals (n = 200) presenting at the reference laboratory in Dhaka, Bangladesh. The specimens were subjected to rapid HIV tests using the national algorithm with A1 = Alere Determine (United States), A2 = Uni-Gold (Ireland), and A3 = First Response (India). The sensitivity and specificity of the test results, and the operational cost were compared with current serum-based testing. Results The sensitivities [95% of confidence interval (CI)] for A1, A2, and A3 tests using whole blood were 100% (CI: 99.1–100%), 100% (CI: 99.1–100%), and 97% (CI: 96.4–98.2%), respectively, and specificities of all test kits were 100% (CI: 99.1–100%). Significant (P < 0.05) reduction in the cost of establishing HTC centre and consumables by 94 and 61%, respectively, were observed. The cost of administration and external quality assurance reduced by 39 and 43%, respectively. Overall, there was a 36% cost reduction in total operational cost of rapid HIV testing with blood when compared with serum. Conclusion Considering the similar sensitivity and specificity of the two specimens, and significant cost reduction, rapid HIV testing with whole blood is feasible. A review of the national HIV rapid testing algorithm with whole blood will contribute toward improving HTC coverage in Bangladesh. PMID:26945143

  19. Rapid and simple method for serotyping of staphylocoagulase using polystyrene latex particles.

    PubMed

    Kouguchi, Yoshihiro; Fujiwara, Takako; Teramoto, Miki

    2009-01-01

    The serotyping of staphylocoagulase is widely used in Japan. However, the conventional immunoassay based on neutralization of the antisera is so laborious and time-consuming that it is not widely used in the other countries. In order to overcome these drawbacks we developed a novel staphylocoagulase serotyping method based on a microplate format using polystyrene latex particles. Addition of latex particles promotes the formation of fibrin complexes, which represents a more rapidly and easily detected endpoint. For 83 strains, 90% were classified into serotypes within 3 h, and there was no discrepancy in the results between our method and the conventional method. These results indicate that the present microplate method is rapid, simple, and interpretable. PMID:20528093

  20. Simple model for rapidity fluctuations in the initial state of ultrarelativistic heavy-ion collisions

    NASA Astrophysics Data System (ADS)

    Broniowski, Wojciech; BoŻek, Piotr

    2016-06-01

    Two-particle pseudorapidity correlations are analyzed in a simple model, where in the initial stage of the reaction multiple sources, extended in rapidity, are created. We show how the fluctuations of the length of the sources in rapidity generate correlations in the initial entropy deposition, which later contribute to the observed longitudinal correlations in hadron production. Our analysis, which is analytic and leads to straightforward formulas, allows us to understand the structure of the correlations, in particular to identify the component related to the fluctuation of the numbers of sources and the component from the length fluctuations. We also present the results in terms of the expansion in the basis of the Legendre polynomials. A number of further effects are discussed, such as smearing of the pseudorapidity distributions or resonance decays. Our results reproduce qualitatively and semiquantitatively the basic features of the recent measurements at the LHC.

  1. A simple, low-cost staining method for rapid-throughput analysis of tumor spheroids

    PubMed Central

    Eckerdt, Frank; Alvarez, Angel; Bell, Jonathan; Arvanitis, Constadina; Iqbal, Asneha; Arslan, Ahmet D.; Hu, Bo; Cheng, Shi-Yuan; Goldman, Stewart; Platanias, Leonidas C.

    2016-01-01

    Tumor spheroids are becoming an important tool for the investigation of cancer stem cell (CSC) function in tumors; thus, low-cost and high-throughput methods for drug screening of tumor spheroids are needed. Using neurospheres as non-adherent three-dimensional (3-D) cultures, we developed a simple, low-cost acridine orange (AO)–based method that allows for rapid analysis of live neurospheres by fluorescence microscopy in a 96-well format. This assay measures the cross-section area of a spheroid, which corresponds to cell viability. Our novel method allows rapid screening of a panel of anti-proliferative drugs to assess inhibitory effects on the growth of cancer stem cells in 3-D cultures. PMID:26757811

  2. Simple and rapid multiplex PCR for identification of the main human diarrheagenic Escherichia coli.

    PubMed

    Tobias, Joshua; Vutukuru, Sreekanth-Reddy

    2012-10-12

    Establishment of a simple and rapid multiplex PCR system for identification of the main diarrheagenic E. coli categories, including enteroaggregative E. coli, enterotoxigenic E. coli, enteropathogenic E. coli, and enterohemorrhagic E. coli, is described. This two-step multiplex PCR system allows the identification by targeting CVD432, LT, STh, STp, Eae, Bfp, Stx1, and Stx2. By applying the developed multiplex PCR system, categorization of E. coli isolates isolated from stool samples of infants with diarrhea into the main diarrheagenic E. coli categories is also shown. PMID:22192837

  3. Cellufine sulfate column chromatography as a simple, rapid, and effective method to purify dengue virus.

    PubMed

    Kanlaya, Rattiyaporn; Thongboonkerd, Visith

    2016-08-01

    Conventional method to purify/concentrate dengue virus (DENV) is time-consuming with low virus recovery yield. Herein, we applied cellufine sulfate column chromatography to purify/concentrate DENV based on the mimicry between heparan sulfate and DENV envelope protein. Comparative analysis demonstrated that this new method offered higher purity (as determined by less contamination of bovine serum albumin) and recovery yield (as determined by greater infectivity). Moreover, overall duration used for cellufine sulfate column chromatography to purify/concentrate DENV was approximately 1/20 of that of conventional method. Therefore, cellufine sulfate column chromatography serves as a simple, rapid, and effective alternative method for DENV purification/concentration. PMID:27155240

  4. Spatial and social inequities in HIV testing utilization in the context of rapid scale-up of HIV/AIDS services in rural Mozambique

    PubMed Central

    Yao, Jing; Agadjanian, Victor; Murray, Alan T.

    2015-01-01

    The massive scale-up of HIV counseling, testing, and treatment services in resource-limited sub-Saharan settings with high HIV prevalence has significant implications for the course of the HIV/AIDS epidemic. It also offers important broader policy lessons for improving access to critical health services. Applying GIS-based methods and multilevel regression analysis to unique longitudinal three-wave survey data from rural Mozambique, this study investigates the impact of a rapid expansion of HIV-related services on access to and utilization of HIV testing. The results illustrate the declining importance of spatial barriers to utilization of HIV testing services as these services expanded. In addition, the expansion of HIV-related services decreased the spatial variability of HIV testing among the survey respondents. At the same time, some important non-spatial variation, such as that in educational level, persisted despite the expansion of services. These results illustrate the process and consequences of health service diffusion. PMID:24835024

  5. Spatial and social inequities in HIV testing utilization in the context of rapid scale-up of HIV/AIDS services in rural Mozambique.

    PubMed

    Yao, Jing; Agadjanian, Victor; Murray, Alan T

    2014-07-01

    The massive scale-up of HIV counseling, testing, and treatment services in resource-limited sub-Saharan settings with high HIV prevalence has significant implications for the course of the HIV/AIDS epidemic. It also offers important broader policy lessons for improving access to critical health services. Applying GIS-based methods and multilevel regression analysis to unique longitudinal three-wave survey data from rural Mozambique, this study investigates the impact of a rapid expansion of HIV-related services on access to and utilization of HIV testing. The results illustrate the declining importance of spatial barriers to utilization of HIV testing services as these services expanded. In addition, the expansion of HIV-related services decreased the spatial variability of HIV testing among the survey respondents. At the same time, some important non-spatial variation, such as that in educational level, persisted despite the expansion of services. These results illustrate the process and consequences of health service diffusion. PMID:24835024

  6. An aptamer-based dipstick assay for the rapid and simple detection of aflatoxin B1.

    PubMed

    Shim, Won-Bo; Kim, Min Jin; Mun, Hyoyoung; Kim, Min-Gon

    2014-12-15

    A rapid and simple dipstick assay based on an aptamer has been developed for the determination of aflatoxin B1 (AFB1). The dipstick assay format was based on a competitive reaction of the biotin-modified aptamer specific to AFB1 between target and cy5-modified DNA probes. Streptavidin and anti-cy5 antibody as capture reagents were immobilized at test and control lines on a membrane of the dipstick assay. After optimization, the limit of detection for the dipstick assay was 0.1 ng/ml AFB1 in buffer. The method was confirmed to be specific to AFB1, and the entire process of the assay can be completed within 30 min. Aqueous methanol (20%) provided a good extraction efficiency, and the matrix influence from corn extracts was successfully reduced through 2-fold dilution. The results of AFB1 analysis for corn samples spiked with known concentration of AFB1 by the dipstick assay and ELISA showed good agreement. The cut-off value of the dipstick assay for corn samples was 0.3 ng/g AFB1. Therefore, the dipstick assay is first reported and considered as a rapid, simple, on-site and inexpensive screening tool for AFB1 determination in grains as well as a corn. PMID:25032679

  7. Simple and rapid detection of Tilletia horrida causing rice kernel smut in rice seeds.

    PubMed

    Chen, Yu; Yang, Xue; Yao, Jian; Kyaw, Ei Phyu; Zhang, Ai-Fang; Li, Yun-Fei; Gu, Chun-Yan; Zang, Hao-Yu; Gao, Tong-Chun

    2016-01-01

    A simple and rapid method for the detection of Tilletia horrida, the causal agent of rice kernel smut, in rice seeds is developed based on specific polymerase chain reaction (PCR). To design the specific primers for the detection of T. horrida, partial sequences of internal transcribed spacer (ITS) DNA region of T. horrida, T. controversa, T. walkeri, T. ehrhartae, T. indica and T. caries were analyzed and compared. A 503-bp fragment was amplified with the designed primers from the T. horrida genomic DNA. However, no PCR product was obtained from the DNA of other five Tilletia species and 22 fungal plant pathogens tested in the present work indicating the specificity of the primers for the detection of T. horrida. The PCR was performed by directly using the spores, isolated from the 21 different rice seed samples, as template DNA. The T. horrida was detected in 6 of the samples, indicating that 28.6% of the rice samples were contaminated with the kernel smut pathogen. This simple PCR based diagnostic assay can be applied for the direct and rapid detection and identification of T. horrida to screen large numbers of rice seed samples. PMID:27624858

  8. Acceptability of rapid oral fluid HIV testing among male injection drug users in Taiwan, 1997 and 2007.

    PubMed

    Lyu, Shu-Yu; Morisky, Donald E; Yeh, Ching-Ying; Twu, Shiing-Jer; Peng, Eugene Yu-Chang; Malow, Robert M

    2011-04-01

    Rapid oral fluid HIV testing (rapid oral testing) is in the process of being adapted in Taiwan and elsewhere given its advantages over prior HIV testing methods. To guide this process, we examined the acceptability of rapid oral testing at two time points (i.e., 1997 and 2007) among one of the highest risk populations, male injection drug users (IDUs). For this purpose, an anonymous self-administered survey was completed by HIV-negative IDUs involved in the criminal justice system in 1997 (N (1)=137 parolees) and 2007 (N (2)=106 prisoners). A social marketing model helped guide the design of our questionnaire to assess the acceptability of rapid oral testing. This included assessing a new product, across four marketing dimensions: product, price, promotion, and place. Results revealed that in both 1997 and 2007, over 90% indicated that rapid oral testing would be highly acceptable, particularly if the cost was under US$6, and that a pharmacy would be the most appropriate and accessible venue for selling the rapid oral testing kits. The vast majority of survey respondents believed that the cost of rapid oral testing should be federally subsidized and that television and newspaper advertisements would be the most effective media to advertise for rapid oral testing. Both the 1997 and 2007 surveys suggested that rapid oral HIV testing would be particularly accepted in Taiwan by IDUs after release from the criminal justice system. PMID:21271392

  9. A Simple Paper-Based Colorimetric Device for Rapid Mercury(II) Assay.

    PubMed

    Chen, Weiwei; Fang, Xueen; Li, Hua; Cao, Hongmei; Kong, Jilie

    2016-01-01

    Contamination of the environment by mercury(II) ions (Hg(2+)) poses a serious threat to human health and ecosystems. Up to now, many reported Hg(2+) sensors require complex procedures, long measurement times and sophisticated instrumentation. We have developed a simple, rapid, low cost and naked-eye quantitative method for Hg(2+) environmental analysis using a paper-based colorimetric device (PCD). The sample solution to which platinum nanoparticles (PtNPs) have been added is dispensed to the detection zone on the PCD, where the 3,3,5,5-tetramethylbenzidine (TMB) substrate has been pre-loaded. The PtNPs effect a rapid oxidization of TMB, inducing blue colorization on the PCD. However, Hg(2+) in the solution rapidly interact with the PtNPs, suppressing the oxidation capacity and hence causing a decrease in blue intensity, which can be observed directly by the naked eye. Moreover, Hg(2+) at concentrations as low as 0.01 uM, can be successfully monitored using a fiber optic device, which gives a digital readout proportional to the intensity of the blue color change. This paper-based colorimetric device (PCD) shows great potential for field measurement of Hg(2+). PMID:27554633

  10. A Simple Paper-Based Colorimetric Device for Rapid Mercury(II) Assay

    PubMed Central

    Chen, Weiwei; Fang, Xueen; Li, Hua; Cao, Hongmei; Kong, Jilie

    2016-01-01

    Contamination of the environment by mercury(II) ions (Hg2+) poses a serious threat to human health and ecosystems. Up to now, many reported Hg2+ sensors require complex procedures, long measurement times and sophisticated instrumentation. We have developed a simple, rapid, low cost and naked-eye quantitative method for Hg2+ environmental analysis using a paper-based colorimetric device (PCD). The sample solution to which platinum nanoparticles (PtNPs) have been added is dispensed to the detection zone on the PCD, where the 3,3,5,5-tetramethylbenzidine (TMB) substrate has been pre-loaded. The PtNPs effect a rapid oxidization of TMB, inducing blue colorization on the PCD. However, Hg2+ in the solution rapidly interact with the PtNPs, suppressing the oxidation capacity and hence causing a decrease in blue intensity, which can be observed directly by the naked eye. Moreover, Hg2+ at concentrations as low as 0.01 uM, can be successfully monitored using a fiber optic device, which gives a digital readout proportional to the intensity of the blue color change. This paper-based colorimetric device (PCD) shows great potential for field measurement of Hg2+. PMID:27554633

  11. Cross-sectional study of community serostatus to highlight undiagnosed HIV infections with oral fluid HIV-1/2 rapid test in non-conventional settings.

    PubMed

    Parisi, Maria Rita; Soldini, Laura; Vidoni, Gianmarino; Clemente, Felice; Mabellini, Chiara; Belloni, Teresa; Nozza, Silvia; Brignolo, Livia; Negri, Silvia; Rusconi, Stefano; Schlusnus, Karin; Dorigatti, Fernanda; Lazzarin, Adriano

    2013-04-01

    The submerged portion of undiagnosed HIV infection in Italy is about 30% of subjects found seropositive. This fact represents one of the most important public health problems hindering the control of infection progression. This means we need to fight unawareness and social stigma and promote easy and friendly access to HIV test. We developed a Prevention Program called “EASY test Project”, offering a new rapid HIV test on oral fluid, to evaluate the acceptability of an alternative, free and anonymous test available in different settings (on board a “Motor Home” at public events, Points of Care, STDs outpatient prevention units and GP surgeries). From December 2008 to December 2012 we performed 7,865 HIV saliva tests, with 50 new infections found (0.6% of the total) out of 140,000 informed subjects. From the self-reported characteristics of respondents, the population approaching the EAST test project was represented by males (70%) aged between 20 and 50 years, 61% with a medium-high education level, 62% homosexuals (MSM), 88% reported unsafe sexual behaviours, and 48% had never undergone an HIV screening test. In five years of the Prevention Program, 100% of subjects interviewed gave a general favorable consent in approaching rapid and not invasive screening, immediate return of the result, and a timely specialized approach and treatment of HIV positive subjects. Results from our study confirm that the rapid and alternative test may contribute to HIV prevention strategies and to the control of the spread of infection and HIV disease progression by reaching a larger population, particularly when and where regular screening procedures are difficult to obtain or are not preferred. PMID:23686118

  12. Accuracy in HIV Rapid Testing among Laboratory and Non-laboratory Personnel in Zambia: Observations from the National HIV Proficiency Testing System

    PubMed Central

    Mwangala, Sheila; Musonda, Kunda G.; Monze, Mwaka; Musukwa, Katoba K.; Fylkesnes, Knut

    2016-01-01

    Background Despite rapid task-shifting and scale-up of HIV testing services in high HIV prevalence countries, studies evaluating accuracy remain limited. This study aimed to assess overall accuracy level and factors associated with accuracy in HIV rapid testing in Zambia. Methods Accuracy was investigated among rural and urban HIV testing sites participating in two annual national HIV proficiency testing (PT) exercises conducted in 2009 (n = 282 sites) and 2010 (n = 488 sites). Testers included lay counselors, nurses, laboratory personnel and others. PT panels of five dry tube specimens (DTS) were issued to testing sites by the national reference laboratory (NRL). Site accuracy level was assessed by comparison of reported results to the expected results. Non-parametric rank tests and multiple linear regression models were used to assess variation in accuracy between PT cycles and between tester groups, and to examine factors associated with accuracy respectively. Results Overall accuracy level was 93.1% (95% CI: 91.2–94.9) in 2009 and 96.9% (95% CI: 96.1–97.8) in 2010. Differences in accuracy were seen between the tester groups in 2009 with laboratory personnel being more accurate than non-laboratory personnel, while in 2010 no differences were seen. In both PT exercises, lay counselors and nurses had more difficulties interpreting results, with more occurrences of false-negative, false-positive and indeterminate results. Having received the standard HIV rapid testing training and adherence to the national HIV testing algorithm were positively associated with accuracy. Conclusion The study showed an improvement in tester group and overall accuracy from the first PT exercise to the next. Average number of incorrect test results per 1000 tests performed was reduced from 69 to 31. Further improvement is needed, however, and the national HIV proficiency testing system seems to be an important tool in this regard, which should be continued and needs to be urgently

  13. Costs of Rapid HIV Screening in an Urban Emergency Department and a Nearby County Jail in the Southeastern United States

    PubMed Central

    Spaulding, Anne C.; MacGowan, Robin J.; Copeland, Brittney; Shrestha, Ram K.; Bowden, Chava J.; Kim, Min J.; Margolis, Andrew; Mustaafaa, Genetha; Reid, Laurie C.; Heilpern, Katherine L.; Shah, Bijal B.

    2015-01-01

    Emergency departments and jails provide medical services to persons at risk for HIV infection and are recommended venues for HIV screening. Our main objective in this study was to analyze the cost per new HIV diagnosis associated with the HIV screening program in these two venues. The emergency department’s parallel testing program was conducted at Grady Memorial Hospital in Atlanta, Georgia starting in 2008; the jail’s integrated testing program began at the Fulton County (GA) Jail in 2011. The two sites, four miles apart from one another, employed the same rapid HIV test. Ascertainment that cases were new differed by site; only the jail systematically checked identities against health department HIV registries. The program in the emergency department used dedicated HIV test counselors and made 242 diagnoses over a 40-month period at a cost of $2,981 per diagnosis. The jail program used staff nurses, and found 41 new HIV cases over 10.5 months at a cost of $6,688 per new diagnosis. Differences in methods for ascertainment of new diagnoses, previously undiagnosed HIV sero-positivity, and methodologies used for assessing program costs prevent concluding that one program was more economical than the other. Nonetheless, our findings show that testing in both venues yielded many new diagnoses, with the costs within the range reported in the literature. PMID:26053140

  14. Black men who have sex with men, sexual risk-taking, and willingness to use rapid home HIV tests.

    PubMed

    Eaton, Lisa A; Driffin, Daniel D; Smith, Harlan; Conway-Washington, Christopher; White, Denise; Cherry, Chauncey

    2015-02-01

    The availability of rapid home-based HIV testing (RHT) in the USA has provided us with a valuable, new option in our efforts to identify more people living with HIV and to do so sooner. Furthermore, it is possible that RHT will be or is currently being used as a means of learning one's own and one's partner's HIV status prior to engaging in condomless intercourse. Data regarding knowledge and willingness to use RHT, however, is very limited. In particular, no studies have investigated RHT use among Black men who have sex with men (BMSM). Understanding RHT use among BMSM is critical as we have observed alarming rates of HIV prevalence among this group, and RHT may provide an opportunity to slow HIV transmission among BMSM. In order to better understand RHT, we assessed knowledge, willingness to use and actual use of RHT, HIV testing history, substance use, and sexual risk-taking among 387 HIV-negative BMSM and 157 HIV-positive BMSM attending a community event in the southeastern USA. We used generalized linear modeling to assess factors associated with their willingness to use RHT. Although familiarity with the availability of RHT was somewhat limited among these men, a substantial portion of BMSM did report an interest in using RHT, including with their sex partners. Among HIV-negative BMSM, however, we found a negative relationship between willingness to use RHT and sexual risk-taking, i.e., higher numbers of condomless anal sex acts were associated with a reduction in willingness to use RHT. It appears that men who report the greatest risk-taking for HIV are least interested in RHT. Future research should focus on better understanding concerns regarding RHT among at-risk HIV-negative men and should investigate the usefulness of using RHT as a HIV prevention method. PMID:24906999

  15. Willingness to Use the Oral Fluid HIV Rapid Test among Men Who Have Sex with Men in Beijing, China

    PubMed Central

    Li, Dongliang; Liu, Yingjie; Pan, Stephen W.; Qi, Xiao; Wang, Bo; Luo, Fengji; Xiao, Dong; Shao, Yiming; Ruan, Yuhua

    2013-01-01

    Background Early detection of HIV infection enables timely care and treatment. However, many men who have sex with men (MSM) remain unaware of their HIV status because they do not or are unable to access HIV testing services. Oral fluid HIV rapid tests have the potential to increase HIV testing. This study is the first to evaluate willingness to use the oral fluid test among MSM in China. Methods A cross-sectional study was conducted in Beijing from July to October, 2012. Data were collected by self-administered questionnaires. Results Of 262 who participated in the survey, 223(85.1%) reported that they were willing to use the oral fluid HIV rapid test. Willingness to use the oral fluid test was associated with higher education (adjusted odds ratio (AOR): 2.40, 95% confidence interval (CI): 1.13–5.10), lack of unprotected anal intercourse (UAI) with male partners in the past one month (AOR: 2.38; 95% 95%CI: 1.15–4.95), having taken more than 4 HIV tests (AOR: 3.54; 95%CI:1.52–8.28), and having ever heard of the oral fluid HIV rapid test from gay friends or gay organizations (AOR: 3.24, 95%CI: 1.40–7.51). Among those who expressed willingness to use the oral fluid HIV rapid test, the median amount of money they were willing to pay was 8 dollars. Among the 39 participants who were unwilling to use the oral fluid test, 79.5% (31/39) expressed concerns about the accuracy of the oral fluid HIV rapid test results and 17.9%(7/39) reported that they were not familiar with the oral fluid test and did not know how to use such a test. Conclusions A high proportion of MSM in Beijing appear to be willing to use the oral fluid HIV rapid test. Appropriate cost and education measures could help improve acceptance of the oral fluid test. PMID:23717645

  16. Rapid assessment of HIV risk behavior in drug using sex workers in three cities in South Africa.

    PubMed

    Parry, Charles D H; Dewing, Sarah; Petersen, Petal; Carney, Tara; Needle, Richard; Kroeger, Karen; Treger, Latasha

    2009-10-01

    A rapid assessment was undertaken with drug using commercial sex workers (CSWs) to investigate practices putting them at risk for contracting HIV. It included key informant (KI) (N = 67) and focus group (N = 10) interviews in locations with a high prevalence of drug use in Cape Town, Durban and Pretoria, South Africa. HIV testing of KIs was conducted. Cocaine, Ecstasy, heroin and methaqualone are used by CSWs prior to, during and after sex. Drugs enhance the sexual experience and prolong sex sessions. Interviews revealed inconsistent condom use among CSWs together with other risky sexual practices such as needle sharing. Among CSWs who agreed to HIV testing, 34% tested positive. Barriers to accessing drug treatment and HIV treatment and preventive services were identified. Interventions recognizing the role of drug abuse in HIV transmission should be prioritized, and issues of access to services, stigma and power relations must be considered. PMID:18324470

  17. Clinical evaluation of a simple, rapid procedure for the presumptive identification of anaerobic bacteria.

    PubMed Central

    Holland, J W; Gagnet, S M; Lewis, S A; Stauffer, L R

    1977-01-01

    A simple, rapid procedure for the presumptive identification of anaerobic bacteria has been evaluated. Two hundred and thirty-five clinical isolates were identified using gas-liquid chromatography and 3-ml volumes of a few selected test media. These test media were stored aerobically and incubated in GasPak anaerobic jars. The average incubation time was 39 h. This procedure, when compared to the results of our standard identification procedure, correctly identified 98% of the isolates to the genus level, 83% to the species level, and 83% of Bacteroides fragilis and Bacteroides melaninogenicus to the subspecies level. Fifty-three of the isolates were also identified by using 0.5-ml volumes of test media stored, inoculated, and incubated in an anaerobic glove box. The 3-ml-and the 0.5-ml-volume procedures correctly identified comparable percentages of the 53 isolates. PMID:323283

  18. Measuring acoustic energy density in microchannel acoustophoresis using a simple and rapid light-intensity method.

    PubMed

    Barnkob, Rune; Iranmanesh, Ida; Wiklund, Martin; Bruus, Henrik

    2012-07-01

    We present a simple and rapid method for measuring the acoustic energy density in microchannel acoustophoresis based on light-intensity measurements of a suspension of particles. The method relies on the assumption that each particle in the suspension undergoes single-particle acoustophoresis. It is validated by the single-particle tracking method, and we show by proper re-scaling that the re-scaled light intensity plotted versus re-scaled time falls on a universal curve. The method allows for analysis of moderate-resolution images in the concentration range encountered in typical experiments, and it is an attractive alternative to particle tracking and particle image velocimetry for quantifying acoustophoretic performance in microchannels. PMID:22522812

  19. A rapid and simple method to draw polyethylene nanofibers with enhanced thermal conductivity

    NASA Astrophysics Data System (ADS)

    Ma, Jian; Zhang, Qian; Zhang, Yin; Zhou, Lei; Yang, Juekuan; Ni, Zhonghua

    2016-07-01

    We report on a rapid and simple method to fabricate polyethylene (PE) nanofibers by one-step drawing from PE solution. The diameter of the fiber prepared with this method can be as small as 40 nm. The thermal conductivity of the drawn PE nanofiber was measured with suspended microdevices, and the highest value obtained is 8.8 W m-1 K-1, which is very close to that of electrospun PE nanofibers, and over 20 times higher than bulk value. Raman spectra of these drawn PE nanofibers indicate that molecular chains in these fibers can be as well aligned as that in electrospun fibers, which results in the enhanced thermal conductivity of the drawn PE nanofibers.

  20. mRNA 5'-cap binding activity in purified influenza virus detected by simple, rapid assay.

    PubMed Central

    Kroath, H; Shatkin, A J

    1982-01-01

    Reovirus mRNA 5'-terminal caps were 3'-radiolabeled with pCp and as affinity probes for proteins with cap binding activity. A rapid, simple, and sensitive blot assay was devised that could detect cellular cap binding protein in a complex polypeptide mixture. By using this method, cap binding activity was found in detergent-treated influenza virus but not in reovirus or vaccinia virus. Preincubation of capped reovirus mRNA with purified cellular cap binding protein reduced its primer effect on influenza transcriptase, whereas priming by ApG was not affected. The results indicate that influenza transcriptase complexes include cap-recognizing proteins that are involved in the formation of chimeric mRNAs. Images PMID:7097854

  1. Nanoparticle-based energy transfer for rapid and simple detection of protein glycosylation

    SciTech Connect

    Oh, Eunkeu; Lee, Dohoon; Kim, Young-Pil; Cha, Seung YOUP; Oh, Doo BEYONG; Kim, Jungbae; Kang, Hyun AH; Kim, Hak SUNG

    2006-12-04

    Glycan moiety of glycoproteins plays an essential role in its biological activity in vivo, and the analysis of glycosylation is of great importance in the development of protein therapeutics. In this study, we report a rapid and simple detection of protein glycosylation based on the fluorescence resonance energy transfer (FRET) between concanavalin A-conjugated gold nanoparticles (ConA-AuNPs) and dextran-conjugated quantum dots (Dex-QDs). The increased photoluminescence (PL) signals of Dex-QDs due to the competitive inhibition of glycoproteins were well correlated with the glycosylation chain length of glucose oxidases as well as the mannosylation degree of bovine serum albumin (BSA). The parallel analysis of the diversely mannosylated BSAs using an image analyzer further demonstrated the potential of this new technique in high-throughput screening of glycoprotein and carbohydrate therapeutics.

  2. Simple and Rapid Synthesis of Magnetite/Hydroxyapatite Composites for Hyperthermia Treatments via a Mechanochemical Route

    PubMed Central

    Iwasaki, Tomohiro; Nakatsuka, Ryo; Murase, Kenya; Takata, Hiroshige; Nakamura, Hideya; Watano, Satoru

    2013-01-01

    This paper presents a simple method for the rapid synthesis of magnetite/hydroxyapatite composite particles. In this method, superparamagnetic magnetite nanoparticles are first synthesized by coprecipitation using ferrous chloride and ferric chloride. Immediately following the synthesis, carbonate-substituted (B-type) hydroxyapatite particles are mechanochemically synthesized by wet milling dicalcium phosphate dihydrate and calcium carbonate in a dispersed suspension of magnetite nanoparticles, during which the magnetite nanoparticles are incorporated into the hydroxyapatite matrix. We observed that the resultant magnetite/hydroxyapatite composites possessed a homogeneous dispersion of magnetite nanoparticles, characterized by an absence of large aggregates. When this material was subjected to an alternating magnetic field, the heat generated increased with increasing magnetite concentration. For a magnetite concentration of 30 mass%, a temperature increase greater than 20 K was achieved in less than 50 s. These results suggest that our composites exhibit good hyperthermia properties and are promising candidates for hyperthermia treatments. PMID:23629669

  3. A simple and rapid vascular anastomosis for emergency surgery: a technical case report

    PubMed Central

    Ball, Chad G; Feliciano, David V

    2009-01-01

    A 22-year old male presented with a transected femoral artery following a gunshot wound. He underwent a successful primary repair following limited segmental resection of the injured segment. End-to-end anastomoses after resection of injured arteries include, but are not limited to, interrupted and continuous suturing with, or without "parachuting" of the graft and/or vessel. We offer a rapid and reliable repair using a conceptually and operationally simple technique. Major advantages include: 1) the operating system is always oriented towards the surgeon, 2) the posterior row of sutures is placed as both ends are readily visualized, avoiding the need for potentially obscuring traction stitches, and 3) flushing is easily performed prior to completing the anterior suture row. PMID:19650926

  4. Dansylation metabolite assay: a simple and rapid method for sample amount normalization in metabolomics.

    PubMed

    Wu, Yiman; Li, Liang

    2014-10-01

    Metabolomics involves the comparison of the metabolomes of individual samples from two or more groups to reveal the metabolic differences. In order to measure the metabolite concentration differences accurately, using the same amount of starting materials is essential. In this work, we describe a simple and rapid method for sample amount normalization. It is based on dansylation labeling of the amine and phenol submetabolome of an individual sample, followed by solvent extraction of the labeled metabolites and ultraviolet (UV) absorbance measurement using a microplate reader. A calibration curve of a mixture of 17 dansyl-labeled amino acid standards is used to determine the total concentration of the labeled metabolites in a sample. According to the measured concentrations of individual samples, the volume of an aliquot taken from each sample is adjusted so that the same sample amount is taken for subsequent metabolome comparison. As an example of applications, this dansylation metabolite assay method is shown to be useful in comparative metabolome analysis of two different E. coli strains using a differential chemical isotope labeling LC-MS platform. Because of the low cost of equipment and reagents and the simple procedure used in the assay, this method can be readily implemented. We envisage that, this assay, which is analogous to the bicinchoninic acid (BCA) protein assay widely used in proteomics, will be applicable to many types of samples for quantitative metabolomics. PMID:25215550

  5. MagaZorb: a simple tool for rapid isolation of viral nucleic acids.

    PubMed

    Nargessi, Dokhi; Ou, Chin-Yih

    2010-04-15

    Effective isolation of nucleic acids from samples containing viral materials is an essential step for accurate diagnosis of viral infections. The necessity of this critical step before analytical identification and diagnosis of viral infections is paramount to screening programs and to identifying and monitoring epidemics and pandemics. With molecular assays rapidly evolving into routine practice, clinical laboratories face several challenges, including presence of small amounts of viral nucleic acids in abundant levels of genomic DNA and total RNA, processing of various sample types, and carry-over of polymerase chain reaction inhibitors, which could significantly affect polymerase chain reaction and microarray results. MagaZorb nucleic acid isolation technology overcomes these challenges and offers a simple and reliable method for isolation of high-quality and high-yield nucleic acids. Although the MagaZorb technology is readily adaptable to automated platforms, it is also well suited to laboratories in remote areas of resource-poor countries, because a simple magnet is the only device required to perform the procedure manually. Performance characteristics and clinical application of the MagaZorb technology are briefly described here. PMID:20225944

  6. Simple and rapid quantification of brominated vegetable oil in commercial soft drinks by LC-MS.

    PubMed

    Chitranshi, Priyanka; Gamboa da Costa, Gonçalo

    2016-12-15

    We report here a simple and rapid method for the quantification of brominated vegetable oil (BVO) in soft drinks based upon liquid chromatography-electrospray ionization mass spectrometry. Unlike previously reported methods, this novel method does not require hydrolysis, extraction or derivatization steps, but rather a simple "dilute and shoot" sample preparation. The quantification is conducted by mass spectrometry in selected ion recording mode and a single point standard addition procedure. The method was validated in the range of 5-25μg/mL BVO, encompassing the legal limit of 15μg/mL established by the US FDA for fruit-flavored beverages in the US market. The method was characterized by excellent intra- and inter-assay accuracy (97.3-103.4%) and very low imprecision [0.5-3.6% (RSD)]. The direct nature of the quantification, simplicity, and excellent statistical performance of this methodology constitute clear advantages in relation to previously published methods for the analysis of BVO in soft drinks. PMID:27451219

  7. HIV rapid testing as a key strategy for prevention of mother-to-child transmission in Brazil

    PubMed Central

    Veloso, Valdiléa G; Bastos, Francisco I; Portela, Margareth Crisóstomo; Grinsztejn, Beatriz; João, Esau Custodio; da Silva Pilotto, Jose Henrique; Araújo, Ana Beatriz Busch; Santos, Breno Riegel; da Fonseca, Rosana Campos; Kreitchmann, Regis; Derrico, Monica; Friedman, Ruth Khalili; Cunha, Cynthia B; Morgado, Mariza Gonçalves; Saines, Karin Nielsen; Bryson, Yvonne J

    2015-01-01

    OBJECTIVE To assess the feasibility of HIV rapid testing for pregnant women at maternity hospital admission and of subsequent interventions to reduce perinatal HIV transmission. METHODS Study based on a convenience sample of women unaware of their HIV serostatus when they were admitted to delivery in public maternity hospitals in Rio de Janeiro and Porto Alegre, Brazil, between March 2000 and April 2002. Women were counseled and tested using the Determine HIV1/2 Rapid Test. HIV infection was confirmed using the Brazilian algorithm for HIV infection diagnosis. In utero transmission of HIV was determined using HIVDNA-PCR. There were performed descriptive analyses of sociodemographic data, number of previous pregnancies and abortions, number of prenatal care visits, timing of HIV testing, HIV rapid test result, neonatal and mother-to-child transmission interventions, by city studied. RESULTS HIV prevalence in women was 6.5% (N=1,439) in Porto Alegre and 1.3% (N=3.778) in Rio de Janeiro. In Porto Alegre most of women were tested during labor (88.7%), while in Rio de Janeiro most were tested in the postpartum (67.5%). One hundred and forty-four infants were born to 143 HIV-infected women. All newborns but one in each city received at least prophylaxis with oral zidovudine. It was possible to completely avoid newborn exposure to breast milk in 96.8% and 51.1% of the cases in Porto Alegre and Rio de Janeiro, respectively. Injectable intravenous zidovudine was administered during labor to 68.8% and 27.7% newborns in Porto Alegre and Rio de Janeiro, respectively. Among those from whom blood samples were collected within 48 hours of birth, in utero transmission of HIV was confirmed in 4 cases in Rio de Janeiro (4/47) and 6 cases in Porto Alegre (6/79). CONCLUSIONS The strategy proved feasible in maternity hospitals in Rio de Janeiro and Porto Alegre. Efforts must be taken to maximize HIV testing during labor. There is a need of strong social support to provide this

  8. Laboratory Evaluation of a Dual-Path Platform Assay for Rapid Point-of-Care HIV and Syphilis Testing.

    PubMed

    Leon, S R; Ramos, L B; Vargas, S K; Kojima, N; Perez, D G; Caceres, C F; Klausner, J D

    2016-02-01

    We assessed the laboratory performance of the Chembio dual-path platform HIV-syphilis rapid immunodiagnostic test and electronic reader for detection of HIV and Treponema pallidum antibodies in 450 previously characterized serum specimens. For visual or electronic reader HIV antibody detection, the sensitivity was 100% and the specificity was 98.7%. For visual T. pallidum antibody detection, the test sensitivity was 94.7% and the specificity was 100.0%; with the electronic reader, the sensitivity was 94.7% and the specificity was 99.7%. PMID:26659215

  9. A novel RT-LAMP assay for rapid and simple detection of classical swine fever virus.

    PubMed

    Chen, Lei; Fan, Xue-zheng; Wang, Qin; Xu, Lu; Zhao, Qi-zu; Zhou, Yuan-chen; Liu, Jun; Tang, Bo; Zou, Xing-qi

    2010-02-01

    A simple and rapid assay for the detection of Classical swine fever virus (CSFV) was established using reverse transcription loop-mediated isothermal amplification (RT-LAMP). This study describes the amplification of the genomic RNA of CSFV under isothermal conditions (63 °C) within one hour, using a set of six primers (two outer primers, two inner primers and two loop primers). This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR. This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV. PRRSV. SIV. PRV-PCV, thus showed a good specificity. Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition, either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye. Because RT-LAMP is low-cost and produces rapid results, it has the potential to be an excellent tool for CSFV surveillance in the field, especially in developing countries. PMID:20960285

  10. Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

    PubMed Central

    Rettcher, Stefanie; Jungk, Felicitas; Kühn, Christoph; Krause, Hans-Joachim; Nölke, Greta; Commandeur, Ulrich; Fischer, Rainer; Schillberg, Stefan

    2015-01-01

    Plant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles. Grapevine fanleaf virus (GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 μg/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, including Potato virus X and Tobacco mosaic virus, with detection limits of 2 to 60 ng/ml. PMID:25710366

  11. A Simple and Rapid Identification Method for Mycobacterium bovis BCG with Loop-Mediated Isothermal Amplification

    PubMed Central

    Kouzaki, Yuji; Maeda, Takuya; Sasaki, Hiroaki; Tamura, Shinsuke; Hamamoto, Takaaki; Yuki, Atsushi; Sato, Akinori; Miyahira, Yasushi; Kawana, Akihiko

    2015-01-01

    Bacillus Calmette-Guérin (BCG) is widely used as a live attenuated vaccine against Mycobacterium tuberculosis and is an agent for standard prophylaxis against the recurrence of bladder cancer. Unfortunately, it can cause severe infectious diseases, especially in immunocompromised patients, and the ability to immediately distinguish BCG from other M. tuberculosis complexes is therefore important. In this study, we developed a simple and easy-to-perform identification procedure using loop-mediated amplification (LAMP) to detect deletions within the region of difference, which is deleted specifically in all M. bovis BCG strains. Reactions were performed at 64°C for 30 min and successful targeted gene amplifications were detected by real-time turbidity using a turbidimeter and visual inspection of color change. The assay had an equivalent detection limit of 1.0 pg of genomic DNA using a turbidimeter whereas it was 10 pg with visual inspection, and it showed specificity against 49 strains of 44 pathogens, including M. tuberculosis complex. The expected LAMP products were confirmed through identical melting curves in real-time LAMP procedures. We employed the Procedure for Ultra Rapid Extraction (PURE) kit to isolate mycobacterial DNA and found that the highest sensitivity limit with a minimum total cell count of mycobacterium (including DNA purification with PURE) was up to 1 × 103 cells/reaction, based on color changes under natural light with FDA reagents. The detection limit of this procedure when applied to artificial serum, urine, cerebrospinal fluid, and bronchoalveolar lavage fluid samples was also about 1 × 103 cells/reaction. Therefore, this substitute method using conventional culture or clinical specimens followed by LAMP combined with PURE could be a powerful tool to enable the rapid identification of M. bovis BCG as point-of-care testing. It is suitable for practical use not only in resource-limited situations, but also in any clinical situation

  12. Rapid High-Level Production of Functional HIV Broadly Neutralizing Monoclonal Antibodies in Transient Plant Expression Systems

    PubMed Central

    Rosenberg, Yvonne; Sack, Markus; Montefiori, David; Forthal, Donald; Mao, Lingjun; -Abanto, Segundo Hernandez; Urban, Lori; Landucci, Gary; Fischer, Rainer; Jiang, Xiaoming

    2013-01-01

    Passive immunotherapy using anti-HIV broadly neutralizing monoclonal antibodies (mAbs) has shown promise as an HIV treatment, reducing mother-to-child-transmission (MTCT) of simian/human immunodeficiency virus (SHIV) in non-human primates and decreasing viral rebound in patients who ceased receiving anti-viral drugs. In addition, a cocktail of potent mAbs may be useful as mucosal microbicides and provide an effective therapy for post-exposure prophylaxis. However, even highly neutralizing HIV mAbs used today may lose their effectiveness if resistance occurs, requiring the rapid production of new or engineered mAbs on an ongoing basis in order to counteract the viral resistance or the spread of a certain HIV-1 clade in a particular region or patient. Plant-based expression systems are fast, inexpensive and scalable and are becoming increasingly popular for the production of proteins and monoclonal antibodies. In the present study, Agrobacterium-mediated transient transfection of plants, utilizing two species of Nicotiana, have been tested to rapidly produce high levels of an HIV 89.6PΔ140env and several well-studied anti-HIV neutralizing monoclonal antibodies (b12, 2G12, 2F5, 4E10, m43, VRC01) or a single chain antibody construct (m9), for evaluation in cell-based viral inhibition assays. The protein-A purified plant-derived antibodies were intact, efficiently bound HIV envelope, and were equivalent to, or in one case better than, their counterparts produced in mammalian CHO or HEK-293 cells in both neutralization and antibody dependent viral inhibition assays. These data indicate that transient plant-based transient expression systems are very adaptable and could rapidly generate high levels of newly identified functional recombinant HIV neutralizing antibodies when required. In addition, they warrant detailed cost-benefit analysis of prolonged incubation in plants to further increase mAb production. PMID:23533588

  13. Factors Influencing Uptake of Rapid HIV and Hepatitis C Screening Among Drug Misusing Adult Emergency Department Patients: Implications for Future HIV/HCV Screening Interventions.

    PubMed

    Merchant, Roland C; DeLong, Allison K; Liu, Tao; Baird, Janette R

    2015-11-01

    In this randomized, controlled trial among 957 English- or Spanish-speaking drug misusing adult emergency department (ED) patients, we determined if a tailored brief intervention (BI) increased uptake of rapid HIV/HCV screening, and identified factors associated with greater screening uptake. Rapid HIV/HCV screening uptake was greater in the control than the BI arm (45 vs. 38 %; p < 0.04). Screening uptake depended on elapsed study time and which research staff member offered testing. In the control arm, uptake was lowest for those spending <30 or ≥90 min in the study. In the BI arm, screening uptake generally increased over time. Tailored BI content specifically addressing participant HIV/HCV knowledge, HIV/HCV risk behaviors, or need for HIV/HCV screening was not associated with greater screening uptake. These study findings suggested factors that should be considered when designing future ED-based screening initiatives, such as elapsed study time, who offers testing, and the content of interventions. PMID:26036465

  14. Rapid acid digestion and simple microplate method for milk iodine determination.

    PubMed

    Hedayati, Mehdi; Ordookhani, Arash; Daneshpour, Maryam Sadat; Azizi, Fereidoun

    2007-01-01

    Iodine deficiency leads to deficiency of thyroid hormones, which causes mental retardation in infant. Laboratory confirmation is important in its diagnosis. The major problems associated with the existing methods for iodine determination in milk samples are: 1) nonsafe alkaline solution; 2) harsh thermal condition; and 3) extra time required to complete various steps. In this study, a simple and rapid colorimetric method was investigated, which used acid digestion in combination with a rapid microplate reading format method to determine the total iodine content in milk. Sample digestion was done on 50 microL milk in metavanadate/perchloric, at 230 degrees C for 10 min. After digestion, iodine determination was based on the Sandell-Kolthoff reaction. The reaction results were read in 96-well microplates by an enzyme-linked immunosorbent assay (ELISA) reader. The determination range of the assay was between 2 and 40 microg/dL. The within-run coefficient of variation percent in three levels (3, 12, and 36 microg/dL) ranged from 6.7 to 9.3 and between-run coefficients of variation ranged from 8.6 to 12.3%. The results obtained (n=70) by the optimized method have good correlation with the results of alkaline incineration as a reference method (n=70; r2=0.907; y=0.952x+1.77). Recovery tests for accuracy assessment in six levels from 6.2 to 34.2 microg/dL) were between 91.3 and 113%. This method has enabled us to achieve 0.12 microg/dL sensitivity. The results of this study show that a quick acid digestion combined with mild thermal and low sample volume with a quick reading of assay results were the main advantages of the acid digestion and microplate reading format. PMID:17847102

  15. A Novel and Simple Method for Rapid Generation of Recombinant Porcine Adenoviral Vectors for Transgene Expression

    PubMed Central

    Ma, Jing; Wang, Wenbin; Zhang, Lu; Tikoo, Suresh K.; Yang, Zengqi

    2015-01-01

    Many human (different serotypes) and nonhuman adenovirus vectors are being used for gene delivery. However, the current system for isolating recombinant adenoviral vectors is either time-consuming or expensive, especially for the generation of recombinant non-human adenoviral vectors. We herein report a new and simple cloning approach for the rapid generation of a porcine adenovirus (PAdV-3) vector which shows promise for gene transfer to human cells and evasion of human adenovirus type 5 (HAdV-5) immunity. Based on the final cloning plasmid, pFPAV3-CcdB-Cm, and our modified SLiCE strategy (SLiCE cloning and lethal CcdB screening), the process for generating recombinant PAdV-3 plasmids required only one step in 3 days, with a cloning efficiency as high as 620±49.56 clones/ng and zero background (100% accuracy). The recombinant PAdV-3 plasmids could be successfully rescued in porcine retinal pigment epithelium cells (VR1BL), which constitutively express the HAdV-5 E1 and PAdV-3 E1B 55k genes, and the foreign genes were highly expressed at 24 h after transduction into swine testicle (ST) cells. In conclusion, this strategy for generating recombinant PAdV-3 vectors based on our modified SLiCE cloning system was rapid and cost-efficient, which could be used as universal cloning method for modification the other regions of PAdV-3 genome as well as other adenoviral genomes. PMID:26011074

  16. Simple, rapid and accurate molecular diagnosis of acute promyelocytic leukemia by loop mediated amplification technology

    PubMed Central

    Spinelli, Orietta; Rambaldi, Alessandro; Rigo, Francesca; Zanghì, Pamela; D'Agostini, Elena; Amicarelli, Giulia; Colotta, Francesco; Divona, Mariadomenica; Ciardi, Claudia; Coco, Francesco Lo; Minnucci, Giulia

    2015-01-01

    The diagnostic work-up of acute promyelocytic leukemia (APL) includes the cytogenetic demonstration of the t(15;17) translocation and/or the PML-RARA chimeric transcript by RQ-PCR or RT-PCR. This latter assays provide suitable results in 3-6 hours. We describe here two new, rapid and specific assays that detect PML-RARA transcripts, based on the RT-QLAMP (Reverse Transcription-Quenching Loop-mediated Isothermal Amplification) technology in which RNA retrotranscription and cDNA amplification are carried out in a single tube with one enzyme at one temperature, in fluorescence and real time format. A single tube triplex assay detects bcr1 and bcr3 PML-RARA transcripts along with GUS housekeeping gene. A single tube duplex assay detects bcr2 and GUSB. In 73 APL cases, these assays detected in 16 minutes bcr1, bcr2 and bcr3 transcripts. All 81 non-APL samples were negative by RT-QLAMP for chimeric transcripts whereas GUSB was detectable. In 11 APL patients in which RT-PCR yielded equivocal breakpoint type results, RT-QLAMP assays unequivocally and accurately defined the breakpoint type (as confirmed by sequencing). Furthermore, RT-QLAMP could amplify two bcr2 transcripts with particularly extended PML exon 6 deletions not amplified by RQ-PCR. RT-QLAMP reproducible sensitivity is 10−3 for bcr1 and bcr3 and 10−2 for bcr2 thus making this assay particularly attractive at diagnosis and leaving RQ-PCR for the molecular monitoring of minimal residual disease during the follow up. In conclusion, PML-RARA RT-QLAMP compared to RT-PCR or RQ-PCR is a valid improvement to perform rapid, simple and accurate molecular diagnosis of APL. PMID:25815362

  17. Rapid selection of escape mutants by the first CD8 T cell responses in acute HIV-1 infection

    SciTech Connect

    Korber, Bette Tina Marie

    2008-01-01

    The recent failure of a vaccine that primes T cell responses to control primary HIV-1 infection has raised doubts about the role of CD8+ T cells in early HIV-1 infection. We studied four patients who were identified shortly after HIV-1 infection and before seroconversion. In each patient there was very rapid selection of multiple HIV-1 escape mutants in the transmitted virus by CD8 T cells, including examples of complete fixation of non-synonymous substitutions within 2 weeks. Sequencing by single genome amplification suggested that the high rate of virus replication in acute infection gave a selective advantage to virus molecules that contained simultaneous and gained sequential T cell escape mutations. These observations show that whilst early HIV-1 specific CD8 T cells can act against virus, rapid escape means that these T cell responses are unlikely to benefit the patient and may in part explain why current HIV-1 T cell vaccines may not be protective.

  18. A Simple Electrochemical Method for the Rapid Estimation of Antioxidant Potentials of Some Selected Medicinal Plants

    PubMed Central

    Amidi, Salimeh; Mojab, Faraz; Bayandori Moghaddam, Abdolmajid; Tabib, Kimia; Kobarfard, Farzad

    2012-01-01

    Clinical and Epidemiological studies have shown that a diet rich in fruits and vegetables is associated with a decreased risk of cardiovascular diseases, cancers and other related disorders. These beneficial health effects have been attributed in part to the presence of antioxidants in dietary plants. Therefore screening for antioxidant properties of plant extracts has been one of the interests of scientists in this field. Different screening methods have been reported for the evaluation of antioxidant properties of plant extracts in the literature. In the present research a rapid screening method has been introduced based on cyclic voltammetry for antioxidant screening of some selected medicinal plant extracts. Cyclic Voltammetry of methanolic extracts of seven medicinal plants: Buxus hyrcana, Rumex crispus, Achillea millefolium, Zataria multiflora, Ginkgo biloba, Lippia citriodora and Heptaptera anisoptera was carried out at different scan rates. Based on the interpretation of voltammograms, Rumex crispus, Achillea millefolium and Ginkgo biloba showed higher antioxidant capability than the others while Lippia citriodora contained the highest amount of antioxidants. Cyclic voltammetry is expected to be a simple method for screening antioxidants and estimating the antioxidant activity of foods and medicinal plants. PMID:25317192

  19. Rapid and simple UPLC-MS/MS method for precise phytochelatin quantification in alga extracts.

    PubMed

    Bräutigam, Anja; Wesenberg, Dirk; Preud'homme, Hugues; Schaumlöffel, Dirk

    2010-09-01

    Quantitative phytochelatin (PC) analysis is, due to oxidation sensitivity of the PCs, matrix effects, and time consuming sample preparation, still a challenging analytical task. In this study, a rapid, simple, and sensitive method for precise determination of native PCs in crude extracts of the green alga Chlamydomonas reinhardtii was developed. Algae were exposed 48 h to 70 μM Cd. Coupling of ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry with multi-reaction mode transitions for detection permitted the required short-time, high-resolution separation and detection specificity. Thus, under optimized chromatographic conditions, 10 thiol peptides were baseline-separated within 7 min. Relative detection limits in the nanomolar range in microliter sample volumes were achieved (corresponding to absolute detection limits at femtomole level). Next to glutathione (GSH), the most abundant cadmium-induced PCs in C. reinhardtii, namely CysGSH, PC(2), PC(3), CysPC(2), and CysPC(3), were quantified with high reproducibility at concentrations between 15 and 198 nmol g(-1) fresh weight. The biological variation of PC synthesis of nine independently grown alga cultures was determined to be on average 13.7%. PMID:20632163

  20. Simple, low-cost planar flow casting machine for rapid solidification processing

    NASA Astrophysics Data System (ADS)

    Smith, M. T.; Saletore, M.

    1986-08-01

    The design, fabrication, and operation of a relatively simple low-cost planar flow casting (PFC) machine optimized for small-batch processing were investigated by the Bureau of Mines. Several design features found beneficial to PFC process operation include: a ground nozzle stopper to retain the alloy charge during melting; a remote, large-volume pressure vessel connected to the crucible gas system to reduce temperature-induced pressure fluctuations; and the use of a nested induction coil that allows both the melt charge and the crucible reservoir to be located close to the cooling wheel. The results of several PFC process runs are provided showing typical values of the cooling wheel surface velocity, crucible ejection pressure, and crucible nozzle clearance gap. Examination of the rapidly solidified, Fe-based ribbons for thickness, dimensional uniformity, and atomic structure indicated that good quality glassy ribbon could be produced with proper selection of the controllable process variables. In addition, single-variable linear regression analysis was used to determined the effect of each process variable on the resulting ribbon thickness.

  1. Simple, rapid method for direct isolation of nucleic acids from aquatic environments.

    PubMed Central

    Somerville, C C; Knight, I T; Straube, W L; Colwell, R R

    1989-01-01

    Direct isolation of nucleic acids from the environment may be useful in several respects, including the estimation of total biomass, detection of specific organisms and genes, estimations of species diversity, and cloning applications. We have developed a method that facilitates the concentration of microorganisms from aquatic samples and the extraction of their nucleic acids. Natural water samples of 350 to greater than 1,000 ml are concentrated on a single cylindrical filter membrane (type SVGS01015; Millipore Corp., Bedford, Mass.), and cell lysis and proteolysis are carried out within the filter housing. Crude, high-molecular-weight nucleic acid solutions are then drawn off the filter. These solutions can be immediately analyzed, concentrated, or purified, depending on the intended application. The method is simple, rapid, and economical and provides high-molecular-weight chromosomal DNA, plasmid DNA, and speciated RNAs which comigrate with 5S, 16S, and 23S rRNAs. The methods presented here should prove useful in studying both the ecology and the phylogeny of microbes that resist classical culture methods. Images PMID:2467621

  2. A rapid and simple HPLC method for the analysis of propofol in biological fluids.

    PubMed

    Cussonneau, Xavier; De Smet, Els; Lantsoght, Kristof; Salvi, Jean-Paul; Bolon-Larger, Magali; Boulieu, Roselyne

    2007-07-27

    A selective and sensitive high-performance liquid chromatographic method for the analysis of propofol in biological samples was developed. Propofol and thymol (internal standard) were analysed on a Purospher RP-18 endcapped (75 mmx4 mm, 3 microm) stationary phase using acetonitrile and water (65:35, v/v) as eluents at a flow rate of 0.6 mL/min. The excitation and emission wavelengths were 276 and 310 nm, respectively. Sample treatment consisted of deproteinization by acetonitrile containing the internal standard and direct injection of the supernatant. Mean analytical recovery were 105% (CV 2.0%) at concentrations ranging from 0.05 to 10 mg/L. The quantification limit was 3 ng/mL for a 500 microL sample plasma volume and 5 ng/mL for a 500 microL blood sample. The intra-day and inter-day precisions were lower than 5.5% for three concentrations assessed (0.05, 1.0 and 10.0 mg/L). Considering the column size and the flow rate, the separation was achieved with an analysis time less than 6 min with a reduced consumption of solvent. This rapid HPLC method using a simple treatment procedure is sensitive enough for monitoring propofol in human biological samples. PMID:17129698

  3. A rapid, simple and sensitive flow cytometric system for detection of Plasmodium falciparum.

    PubMed

    Saito-Ito, A; Akai, Y; He, S; Kimura, M; Kawabata, M

    2001-11-01

    We have established a rapid, simple and sensitive flow cytometric system for the detection of Plasmodium falciparum that involves lysing erythrocytes and staining parasites at the same time using a newly developed hemolysing and staining solution containing dodecyl methyl ammonium chloride and acridine orange. In this system, freed parasites of P. falciparum could be plotted separately from erythrocyte ghosts, white blood cells and platelets on the two-dimensional scattergram of forward-angle light scatter and green fluorescence by flow cytometry with an argon laser. It took only 2-3 min per sample to obtain the scattergram and analyze the data, including the time of sample preparation for flow cytometric analysis. Sample preparation with this method does not require any difficult handling procedures. The threshold of parasite detection was almost equal to that of microscopic examination for cultured P. falciparum. The results of drug-susceptibility assays using this system were also almost identical to those obtained using microscopic examination. In this system, parasites at different erythrocytic stages could be easily distinguished. This system must prove useful and practical for basic laboratory studies of P. falciparum including those requiring the differential measurement of parasites at specific erythrocytic stages. PMID:11719111

  4. Hydrodynamic Voltammetry as a Rapid and Simple Method for Evaluating Soil Enzyme Activities

    PubMed Central

    Sazawa, Kazuto; Kuramitz, Hideki

    2015-01-01

    Soil enzymes play essential roles in catalyzing reactions necessary for nutrient cycling in the biosphere. They are also sensitive indicators of ecosystem stress, therefore their evaluation is very important in assessing soil health and quality. The standard soil enzyme assay method based on spectroscopic detection is a complicated operation that requires the removal of soil particles. The purpose of this study was to develop a new soil enzyme assay based on hydrodynamic electrochemical detection using a rotating disk electrode in a microliter droplet. The activities of enzymes were determined by measuring the electrochemical oxidation of p-aminophenol (PAP), following the enzymatic conversion of substrate-conjugated PAP. The calibration curves of β-galactosidase (β-gal), β-glucosidase (β-glu) and acid phosphatase (AcP) showed good linear correlation after being spiked in soils using chronoamperometry. We also performed electrochemical detection using real soils. Hydrodynamic chronoamperometry can be used to assess the AcP in soils, with a detection time of only 90 s. Linear sweep voltammetry was used to measure the amount of PAP released from β-gal and β-glu by enzymatic reaction after 60 min. For the assessment of soil enzymes, the results of hydrodynamic voltammetry assay compared favorably to those using a standard assay procedure, but this new procedure is more user-friendly, rapid and simple. PMID:25746097

  5. Rapid, simple, and cost-effective treatments to achieve long-term hydrophilic PDMS surfaces

    NASA Astrophysics Data System (ADS)

    Hemmilä, Samu; Cauich-Rodríguez, Juan V.; Kreutzer, Joose; Kallio, Pasi

    2012-10-01

    This paper describes rapid, simple, and cost-effective treatments for producing biocompatible and long-term hydrophilic polydimethylsiloxane (PDMS) surfaces identified in an experimental study investigating 39 treatments in all. The wetting of the surfaces was monitored during six months. Changes in surface morphology and chemical composition were also analyzed. Some of the treatments are presented here for the first time, while for earlier presented treatments the selection of investigated parameters was wider and the observation period for the surface wetting longer. The PDMS surfaces were modified by surface activation, physisorption, and synthesis of both “grafting to” and “grafting from” polymer brushes. In surface activation, the PDMS sample was exposed to oxygen plasma, with several combinations of exposure time and RF power. In the physisorption and synthesis of polymer brushes, three commercially available and biocompatible chemicals were used: 2-hydroxyethyl methacrylate (HEMA), polyethylene glycol (PEG), and polyvinylpyrrolidone (PVP). Thirty-three of the 39 treatments rendered the PDMS hydrophilic, and in 12 cases the hydrophilicity lasted at least six months. Seven of these long-term hydrophilic coatings supported a contact angle of 30° or less. Three of the long-lasting hydrophilic coatings required only minutes to prepare.

  6. Rapid and simple isolation of vascular, epidermal and mesophyll cells from plant leaf tissue.

    PubMed

    Endo, Motomu; Shimizu, Hanako; Araki, Takashi

    2016-08-01

    To understand physiological phenomena at the tissue level, elucidation of tissue-specific molecular functions in vivo is required. As an example of the current state of affairs, many genes in plants have been reported to have discordant levels of expression between bulk tissues and the specific tissues in which the respective gene product is principally functional. The principal challenge in deciphering such tissue-specific functions lies in separating tissues with high spatiotemporal resolution to evaluate accurate gene expression profiles. Here, we provide a simple and rapid tissue isolation protocol to isolate all three major leaf tissues (mesophyll, vasculature and epidermis) from Arabidopsis within 30 min with high purity. On the basis of the different cell-to-cell connectivities of tissues, the mesophyll isolation is achieved by making protoplasts, and the vasculature and epidermis isolation is achieved through sonication and enzymatic digestion of leaves. We have successfully tested the protocol on several other plant species, including crop plants such as soybean, tomato and wheat. Furthermore, isolated tissues can be used not only for tissue-specific transcriptome assays but also potentially for tissue-specific proteome and methylome assays. PMID:27388555

  7. ACSM4 polymorphisms are associated with rapid AIDS progression in HIV-infected patients.

    PubMed

    Guzmán-Fulgencio, María; Jiménez, José L; Jiménez-Sousa, María A; Bellón, José M; García-Álvarez, Mónica; Soriano, Vicente; Gijón-Vidaurreta, Paloma; Bernal-Morell, Enrique; Viciana, Pompeyo; Muñoz-Fernández, M Ángeles; Resino, Salvador

    2014-01-01

    : Our aim was to explore the association among ACSM4 and PECI polymorphisms and AIDS progression in 454 HIV-infected patients never treated with antiretroviral drugs (146 long-term nonprogressors, 228 moderate progressors, and 80 rapid progressors). For ACSM4 polymorphisms, rs7137120 AA/AG and rs7961991 CC/CT genotypes had higher odds of having a rapid AIDS progression [odds ratio (OR) = 3.21; 95% of confidence interval (95% CI) = 1.26 to 8.16; P = 0.014 and OR = 3.60; 95% CI = 1.38 to 9.36; P = 0.009, respectively]. Additionally, the ACSM4 haplotype integrated for both rs7961991 A and rs7137120 C alleles had higher odds of having a rapid AIDS progression (OR = 2.85; 95% CI = 1.28 to 6.25; P = 0.010). For PECI polymorphisms, no significant associations were found. In conclusion, ACSM4 polymorphisms might play a significant role in AIDS progression. PMID:23982661

  8. Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris.

    PubMed

    Shah, Kartik A; Clark, John J; Goods, Brittany A; Politano, Timothy J; Mozdzierz, Nicholas J; Zimnisky, Ross M; Leeson, Rachel L; Love, J Christopher; Love, Kerry R

    2015-12-01

    Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria, or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, "hit-to-lead identification" remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigen-specific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV). PMID:26032261

  9. Electrospun solid dispersions of Maraviroc for rapid intravaginal preexposure prophylaxis of HIV.

    PubMed

    Ball, Cameron; Woodrow, Kim A

    2014-08-01

    The development of topical anti-human immunodeficiency virus (HIV) microbicides may provide women with strategies to protect themselves against sexual HIV transmission. Pericoital drug delivery systems intended for use immediately before sex, such as microbicide gels, must deliver high drug doses for maximal effectiveness. The goal of achieving a high antiretroviral dose is complicated by the need to simultaneously retain the dose and quickly release drug compounds into the tissue. For drugs with limited solubility in vaginal gels, increasing the gel volume to increase the dose can result in leakage. While solid dosage forms like films and tablets increase retention, they often require more than 15 min to fully dissolve, potentially increasing the risk of inducing epithelial abrasions during sex. Here, we demonstrate that water-soluble electrospun fibers, with their high surface area-to-volume ratio and ability to disperse antiretrovirals, can serve as an alternative solid dosage form for microbicides requiring both high drug loading and rapid hydration. We formulated maraviroc at up to 28 wt% into electrospun solid dispersions made from either polyvinylpyrrolidone or poly(ethylene oxide) nanofibers or microfibers and investigated the role of drug loading, distribution, and crystallinity in determining drug release rates into aqueous media. We show here that water-soluble electrospun materials can rapidly release maraviroc upon contact with moisture and that drug delivery is faster (less than 6 min under sink conditions) when maraviroc is electrospun in polyvinylpyrrolidone fibers containing an excipient wetting agent. These materials offer an alternative dosage form to current pericoital microbicides. PMID:24913168

  10. Electrospun Solid Dispersions of Maraviroc for Rapid Intravaginal Preexposure Prophylaxis of HIV

    PubMed Central

    Ball, Cameron

    2014-01-01

    The development of topical anti-human immunodeficiency virus (HIV) microbicides may provide women with strategies to protect themselves against sexual HIV transmission. Pericoital drug delivery systems intended for use immediately before sex, such as microbicide gels, must deliver high drug doses for maximal effectiveness. The goal of achieving a high antiretroviral dose is complicated by the need to simultaneously retain the dose and quickly release drug compounds into the tissue. For drugs with limited solubility in vaginal gels, increasing the gel volume to increase the dose can result in leakage. While solid dosage forms like films and tablets increase retention, they often require more than 15 min to fully dissolve, potentially increasing the risk of inducing epithelial abrasions during sex. Here, we demonstrate that water-soluble electrospun fibers, with their high surface area-to-volume ratio and ability to disperse antiretrovirals, can serve as an alternative solid dosage form for microbicides requiring both high drug loading and rapid hydration. We formulated maraviroc at up to 28 wt% into electrospun solid dispersions made from either polyvinylpyrrolidone or poly(ethylene oxide) nanofibers or microfibers and investigated the role of drug loading, distribution, and crystallinity in determining drug release rates into aqueous media. We show here that water-soluble electrospun materials can rapidly release maraviroc upon contact with moisture and that drug delivery is faster (less than 6 min under sink conditions) when maraviroc is electrospun in polyvinylpyrrolidone fibers containing an excipient wetting agent. These materials offer an alternative dosage form to current pericoital microbicides. PMID:24913168

  11. Rapid, simple and efficient method for detection of viral genomes on raspberries.

    PubMed

    Perrin, A; Loutreul, J; Boudaud, N; Bertrand, I; Gantzer, C

    2015-11-01

    In recent years, foodborne viruses, especially human noroviruses (NoV) and hepatitis A virus (HAV), have been increasingly reported as the causes of foodborne disease outbreaks. Soft red fruits, especially raspberries, have a high incidence among the types of food concerned. Due to low infectious doses and low concentrations of enteric viruses in food samples, it is necessary to have an efficient and rapid detection method to implement prevention measures. A standard method for virus detection and quantification in food, including raspberries (XP CEN ISO/TS 15216-1 and -2, 2013) is currently available. This method proposes a consensus detection approach by RT-real time PCR (RT-qPCR) but also a virus extraction procedure based on the elution-concentration principle. In this study, an alternative method of extraction in which RNAs are directly extracted from food matrices (based on direct RNA extraction) has been optimized. First, each step was improved to make it a highly rapid, specific and simple method. Second, the standard virus concentration method was compared with the optimized direct RNA extraction one. Human enteric viral surrogates, Murine Norovirus (MNV) and F-specific RNA bacteriophage GA, were selected according to their adhesion properties and resistance to pH close to our main targets (NoV and HAV). Raspberries were artificially contaminated using two different techniques (immersion and spotting) in order to define a recovery rate and the amounts of virus recovered. Results showed that the direct RNA extraction method revealed significantly higher viral extraction efficiency (46.2%) than the elution-concentration method (20.3%), with similar proportions of inhibitors for both. In the same way with inoculation by spotting, the best recovery rate of GA phage (39.7% against 0.7%) and MNV (42.8% against 0.5%) was observed by direct RNA extraction. For the lowest concentrations of phage and virus in the immersion bath, only the direct RNA extraction method

  12. Simple and rapid in vitro assay for detecting human thyroid peroxidase disruption.

    PubMed

    Jomaa, Barae; de Haan, Laura H J; Peijnenburg, Ad A C M; Bovee, Toine F H; Aarts, Jac M M J G; Rietjens, Ivonne M C M

    2015-01-01

    A simple and rapid luminometric assay for the detection of chemical inhibitors of human thyroid peroxidase (hTPO) activity was developed and validated with 10 model compounds. hTPO was derived from the human thyroid follicular cell line Nthy-ori 3-1 and its activity was quantified by measuring the oxidation of luminol in the presence of hydrogen peroxide (H2O2), which results in the emission of light at 428 nm. In this assay,hTPO activity was shown to be inhibited by 5 known TPO inhibitors and not inhibited by 5 non-inhibitors. Similar results were obtained with porcine TPO (pTPO).The inhibition of hTPO by the model compounds was also tested with guaiacol and Ampliflu Red as alternative indicator substrates. While all substrates allowed the detection of pTPO activity and its inhibition, only the Ampliflu Red and luminol-based methods were sensitive enough to allow the quantification of hTPO activity from Nthy-ori 3-1 cell lysates. Moreover, luminol gave results with a narrower 95% confidence interval and therefore more reliable data.Whole extracts of fast-growing Nthy-ori 3-1 cells circumvent the need for animal-derived thyroid organs,thereby reducing costs, eliminating potential contamination and providing the possibility to study human instead of porcine TPO. Overall, the application of luminol and Nthy-ori 3-1 cell lysate for the detection of the disruption of hTPO activity was found to represent a valuable in vitro alternative and a possible candidate for inclusion within a high throughput integrated testing strategy for the detection of compounds that potentially interfere with normal thyroid function in vivo. PMID:25822105

  13. Exploration of Simple Analytical Approaches for Rapid Detection of Pathogenic Bacteria

    SciTech Connect

    Salma Rahman

    2005-12-17

    Many of the current methods for pathogenic bacterial detection require long sample-preparation and analysis time, as well as complex instrumentation. This dissertation explores simple analytical approaches (e.g., flow cytometry and diffuse reflectance spectroscopy) that may be applied towards ideal requirements of a microbial detection system, through method and instrumentation development, and by the creation and characterization of immunosensing platforms. This dissertation is organized into six sections. In the general Introduction section a literature review on several of the key aspects of this work is presented. First, different approaches for detection of pathogenic bacteria will be reviewed, with a comparison of the relative strengths and weaknesses of each approach, A general overview regarding diffuse reflectance spectroscopy is then presented. Next, the structure and function of self-assembled monolayers (SAMs) formed from organosulfur molecules at gold and micrometer and sub-micrometer patterning of biomolecules using SAMs will be discussed. This section is followed by four research chapters, presented as separate manuscripts. Chapter 1 describes the efforts and challenges towards the creation of imunosensing platforms that exploit the flexibility and structural stability of SAMs of thiols at gold. 1H, 1H, 2H, 2H-perfluorodecyl-1-thiol SAM (PFDT) and dithio-bis(succinimidyl propionate)-(DSP)-derived SAMs were used to construct the platform. Chapter 2 describes the characterization of the PFDT- and DSP-derived SAMs, and the architectures formed when it is coupled to antibodies as well as target bacteria. These studies used infrared reflection spectroscopy (IRS), X-ray photoelectron spectroscopy (XPS), and electrochemical quartz crystal microbalance (EQCM), Chapter 3 presents a new sensitive, and portable diffuse reflection based technique for the rapid identification and quantification of pathogenic bacteria. Chapter 4 reports research efforts in the

  14. A Rapid, Self-confirming Assay for HIV: Simultaneous Detection of Anti-HIV Antibodies and Viral RNA

    PubMed Central

    Chen, Zongyuan; Zhu, Hui; Malamud, Daniel; Barber, Cheryl; Ongagna, Yhombi Yvon Serge; Yasmin, Rubina; Modak, Sayli; Janal, Malvin N.; Abrams, William R.; Montagna, Richard A.

    2016-01-01

    Objective We developed a microfluidic system to simultaneously detect host anti-HIV antibodies and viral RNA in the same specimen in order to satisfy two important diagnostic criteria, especially within resource-limited settings. First, the system can detect acute HIV infection and allow immediate confirmation of a seropositive screening result by detection of HIV RNA. It also addresses the well-known "seroconversion window" during early HIV infection when antibodies are not yet detectable and viral loads are at their highest. Methods We first developed and optimized two separate manual assays for the detection of host anti-HIV antibodies and viral RNA and then converted them to the microfluidic system. We optimized a commercially available serologic assay to run within the microfluidic device while we incorporated the isothermal LAMP assay to detect the presence of viral RNA. The microfluidic device and instrumentation were developed to simultaneously perform both assays without any user intervention. Results The finalized system consists of a disposable injection molded and film-laminated microfluidic CARD disposable device and a portable, software controlled instrument, which together can automatically perform all steps of both assays without any user intervention after the initial loading of samples and reagents. The microfluidic CARD cartridge has multiple microchannels, valves, pumps and reservoirs, which perform the immunoassay, isolates viral RNA for detection by magnetic bead based purification, and Reverse Transcriptase loop-mediated isothermal amplification (RT-LAMP). The microfluidic system was able to detect host anti-HIV antibodies and viral RNA in either a blood or saliva sample. Conclusion The ability to detect antibodies and simultaneously confirm a seropositive HIV-RNA result provides healthcare workers with a complete and accurate appraisal of a patient's infection status in the earliest stages of the disease and represents an important tool for

  15. Brief Report: Relationship and Demographic Factors Associated With Willingness to Use an In-Home Rapid HIV Test to Screen Potential Sex Partners Among a US Sample of HIV-Negative and HIV-Discordant Male Couples.

    PubMed

    Mitchell, Jason W; Sullivan, Patrick S

    2015-06-01

    With dyadic data from a US Internet sample of 275 HIV-negative and 58 discordant male couples, we assessed HIV-negative partnered men's attitudes toward using an in-home rapid HIV test (HT) to screen potential new sex partners and associated factors by multivariate multilevel modeling. HIV-negative partnered men were "likely" to use an HT for screening purposes. More positive attitudes were associated with being in a mixed/nonwhite relationship; having an open sexual agreement. Less positive attitudes were associated with both partners being well educated. These findings may highlight how to make the most of HTs as risk-reduction screening tool among at-risk male couples. PMID:26009834

  16. Relationship and demographic factors associated with willingness to use an in-home HIV rapid test (HT) to screen potential sex partners among a US sample of HIV-negative and HIV-discordant male couples

    PubMed Central

    Mitchell, Jason W.; Sullivan, Patrick S.

    2015-01-01

    With dyadic data from a US Internet sample of 275 HIV-negative and 58 discordant male couples, we assessed HIV-negative partnered men's attitudes towards using an in-home rapid HIV test to screen potential new sex partners and associated factors by multivariate multilevel modeling. HIV-negative partnered men were “likely” to use a HT for screening purposes. More positive attitudes were associated with being in a mixed/nonwhite relationship; having an open sexual agreement. Less positive attitudes were associated with both partners being well educated. These findings may highlight how to make the most of HTs as risk-reduction screening tool among at-risk male couples. PMID:26009834

  17. Simple filter microchip for rapid separation of plasma and viruses from whole blood

    PubMed Central

    Wang, ShuQi; Sarenac, Dusan; Chen, Michael H; Huang, Shih-Han; Giguel, Francoise F; Kuritzkes, Daniel R; Demirci, Utkan

    2012-01-01

    Sample preparation is a significant challenge for detection and sensing technologies, since the presence of blood cells can interfere with the accuracy and reliability of virus detection at the nanoscale for point-of-care testing. To the best of our knowledge, there is not an existing on-chip virus isolation technology that does not use complex fluidic pumps. Here, we presented a lab-on-a-chip filter device to isolate plasma and viruses from unprocessed whole blood based on size exclusion without using a micropump. We demonstrated that viruses (eg, HIV) can be separated on a filter-based chip (2-μm pore size) from HIV-spiked whole blood at high recovery efficiencies of 89.9% ± 5.0%, 80.5% ± 4.3%, and 78.2% ± 3.8%, for viral loads of 1000, 10,000 and 100,000 copies/mL, respectively. Meanwhile, 81.7% ± 6.7% of red blood cells and 89.5% ± 2.4% of white blood cells were retained on 2 μm pore–sized filter microchips. We also tested these filter microchips with seven HIV-infected patient samples and observed recovery efficiencies ranging from 73.1% ± 8.3% to 82.5% ± 4.1%. These results are first steps towards developing disposable point-of-care diagnostics and monitoring devices for resource-constrained settings, as well as hospital and primary care settings. PMID:23055720

  18. Approval and potential use of over-the-counter HIV self-tests: the opinion of participants in a street based HIV rapid testing program in Spain.

    PubMed

    Rosales-Statkus, M Elena; de la Fuente, Luis; Fernández-Balbuena, Sonia; Figueroa, Carmen; Fernàndez-López, Laura; Hoyos, Juan; Ruiz, Mónica; Belza, M José

    2015-03-01

    HIV self-testing, not yet available in Spain, is a strategy thought to be able to increase the number of people tested and testing frequency. 3,373 attenders of a street-based HIV rapid-testing program gave their opinion on authorizing over-the-counter self-tests and a potentially shorter lead time if self-tests were available. 88.0 % of participants were in favor of authorization, 9.2 % had no clear opinion and 2.8 % were against. 54.6 % of men who have sex with men (MSM), 38.4 % of men who have sex with women and 36.3 % of women acknowledged a potential for lead time reduction. Potential lead time was associated with being ≥35 years, having a university degree, never injected drugs, previous HIV tests and being concerned about an HIV positive result, and in MSM, also having ≥5 partners. Self-testing seems a promising strategy for Spain: its authorization is supported by nearly all potential users and almost three quarters of MSM would have opted to advance their current testing if self-tests were available. PMID:25537965

  19. A Multisite Study of the Prevalence of HIV With Rapid Testing in Mental Health Settings

    PubMed Central

    Blank, Michael B.; Himelhoch, Seth S.; Balaji, Alexandra B.; Metzger, David S.; Dixon, Lisa B.; Rose, Charles E.; Oraka, Emeka; Davis-Vogel, Annet; Thompson, William W.; Heffelfinger, James D.

    2014-01-01

    Objectives. We estimated HIV prevalence and risk factors among persons receiving mental health treatment in Philadelphia, Pennsylvania, and Baltimore, Maryland, January 2009 to August 2011. Methods. We used a multisite, cross-sectional design stratified by clinical setting. We tested 1061 individuals for HIV in university-based inpatient psychiatric units (n = 287), intensive case-management programs (n = 273), and community mental health centers (n = 501). Results. Fifty-one individuals (4.8%) were HIV-infected. Confirmed positive HIV tests were 5.9% (95% confidence interval [CI] = 3.7%, 9.4%) for inpatient units, 5.1% (95% CI = 3.1%, 8.5%) for intensive case-management programs, and 4.0% (95% CI = 2.6%, 6.1%) for community mental health centers. Characteristics associated with HIV included Black race, homosexual or bisexual identity, and HCV infection. Conclusions. HIV prevalence for individuals receiving mental health services was about 4 times as high as in the general population. We found a positive association between psychiatric symptom severity and HIV infection, indicating that engaging persons with mental illness in appropriate mental health treatment may be important to HIV prevention. These findings reinforce recommendations for routine HIV testing in all clinical settings to ensure that HIV-infected persons receiving mental health services are identified and referred to timely infectious disease care. PMID:24524493

  20. Clinical versus Rapid Molecular HIV Diagnosis in Hospitalized African Infants: A Randomized Controlled Trial Simulating Point-of-Care Infant Testing

    PubMed Central

    McCollum, Eric D.; Preidis, Geoffrey A.; Maliwichi, Madalitso; Olson, Dan; McCrary, L. Madeline; Kazembe, Peter N.; van der Horst, Charles; Hoffman, Irving; Hosseinipour, Mina C.

    2014-01-01

    Objective Many African infants fail to receive their diagnostic HIV molecular test results and subsequently, antiretroviral therapy (ART). To determine whether a point-of-care molecular HIV test increases ART access for hospitalized Malawian infants, we simulated a point-of-care test using rapid HIV RNA polymerase chain reaction (Rapid PCR) and compared patient outcomes to an optimized standard care that included assessment with the World Health Organization (WHO) clinical algorithm for HIV infection plus a DNA PCR with a turnaround time of several weeks (standard care). Design Randomized controlled trial. Methods Hospitalized HIV-exposed Malawian infants <12 months old were randomized into Rapid PCR or standard care. Rapid PCR infants obtained molecular test results within 48 hours to facilitate immediate ART, similar to a point-of-care test. Standard care infants meeting clinical criteria were also offered inpatient ART. The primary outcome was appropriate in-hospital ART for DNA or RNA PCR-confirmed HIV-infected infants. Results 300 infants were enrolled. A greater proportion of HIV-infected infants receiving Rapid PCR, versus standard care, started inpatient ART (72.3% vs 47.8%, p=0.016). Among molecular test-negative infants, 26.9% receiving standard care unnecessarily initiated inpatient ART, versus 0.0% receiving Rapid PCR (p<0.001). Rapid PCR modestly reduced the median days to ART (3.0 vs 6.5, p=0.001) but did not influence outpatient follow-up for HIV-infected infants (78.1% vs 82.4%, P = 0.418). Conclusions Rapid PCR, versus an optimized standard care, increased the proportion of hospitalized HIV-infected infants initiating ART and reduced ART exposure in molecular test-negative infants, without meaningfully impacting time to ART initiation or follow-up rates. PMID:24326604

  1. Resazurin Microtiter Assay Plate Testing of Mycobacterium tuberculosis Susceptibilities to Second-Line Drugs: Rapid, Simple, and Inexpensive Method

    PubMed Central

    Martin, Anandi; Camacho, Mirtha; Portaels, Françoise; Palomino, Juan Carlos

    2003-01-01

    The emergence of multidrug-resistant tuberculosis calls for new, rapid drug susceptibility tests. We have tested 150 Mycobacterium tuberculosis isolates against the second-line drugs ethionamide, kanamycin, capreomycin, ofloxacin, and para-aminosalicylic acid by the colorimetric resazurin microtiter assay and the proportion method. By visual reading, MICs were obtained after 8 days. A very good correlation between results by the colorimetric resazurin microtiter assay and the proportion method was obtained. The colorimetric resazurin microtiter assay is inexpensive, rapid, and simple to perform, and implementation of the assay is feasible for low-resource countries. PMID:14576129

  2. Simple Adhesive-Tape-Based Sampling of Tomato Surfaces Combined with Rapid Fluorescence In Situ Hybridization for Salmonella Detection▿

    PubMed Central

    Bisha, Bledar; Brehm-Stecher, Byron F.

    2009-01-01

    A simple adhesive-tape-based method for sampling of tomato surfaces was combined with fluorescence in situ hybridization for rapid culture-independent detection of Salmonella strains. Tapes could also be placed face-down on selective agar for on-tape enrichment of captured Salmonella cells. Overlay of cell-charged tapes with small volumes of liquid enrichment media enabled subsequent detection of tape-captured Salmonella via flow cytometry. PMID:19124588

  3. Sex, drugs, and HIV: rapid assessment of HIV risk behaviors among street-based drug using sex workers in Durban, South Africa.

    PubMed

    Needle, Richard; Kroeger, Karen; Belani, Hrishikesh; Achrekar, Angeli; Parry, Charles D; Dewing, Sarah

    2008-11-01

    South Africa is experiencing significant changes in patterns of illicit drug use, including increasing injection and non-injection drug use, and the use of drugs by persons engaged in sex work, both of which could further expand the HIV/AIDS epidemic. In 2005, a rapid ethnographic assessment was conducted in Durban, South Africa, to learn more about patterns of drug use and HIV risk behaviors among drug-using, street-based sex workers. Field teams recruited 52 current injection and non-injection drug users for key informant interviews and focus groups, and they conducted mapping and observation in identified high-risk neighborhoods. Key informants were offered free, voluntary counseling and HIV rapid testing. The results of the assessment indicate that in this population, drugs play an organizing role in patterns of daily activities, with sex work closely linked to the buying, selling, and using of drugs. Participants reported using multiple drugs including crack cocaine, heroin, Ecstasy and Mandrax, and their choices were based on their expectations about the functional role and behavioral and pharmacological properties of the drugs. The organization of sex work and patterns of drug use differ by gender, with males exercising more control over daily routines and drug and sexual transactions than females. Activities of female sex workers are subject to considerable control by individual pimps, many of whom also function as landlords and drug dealers. A strong hold over the overlapping economies of drugs and sex work by a few individuals extends to control of the physical and social settings in which sex is exchanged and drugs are sold and used as well as the terms under which sex work is carried out. The potential for accelerated HIV spread is considerable given the evidence of overlapping drug-using and sexual risk behaviors and the mixing patterns across drug and sexual risk networks. PMID:18678437

  4. Obstetrical, maternal characteristics and outcome of HIV-infected rapid progressor infants at Yaounde: a retrospective study

    PubMed Central

    Dongmo, Roger; Touffic Othman, Carole Leïla; Tatah, Sandra; Njiki Kinkela, Mina Ntoto; Koki Ndombo, Paul Olivier

    2016-01-01

    Background Rapid progressors are exposed to HIV infection at an early stage of life, and the prognosis is poor without treatment. Reducing the proportion of infants who are rapid progressors, require strengthening strategies to achieve the highest level of performance for the PMTCT program. Methods This was a retrospective study carried out on HIV infected infants aged less than 12 months, clinically classified stage 4 (WHO) or having CD4 count <25%. We described maternal and obstetrical characteristics of HIV-infected rapid progressors using univariate and bivariate analysis. Patients’ survival was monitored from the inclusion time to the end of the study. We then estimated their probability of survival with or without anti-retroviral (ARV) treatment from birth using the Kaplan-Meier method. Results The characteristics of the mothers of the 150 rapid progressors infants we included were: low level of education (OR=3.87; P=0.016), CD4 count less than 200/mm3 (OR=43.3; P=0.000), absence of ARV prophylaxis (OR=6.02; P=0.043), or treatment with HAART (OR=5.74; P=0.000) during pregnancy. In the children, the most important findings were lack of co-trimoxazole prophylaxis (OR=11.61; P=0.000) and antiretroviral prophylaxis (OR=2.70; P=0.0344). The survival rate was 84.3% in infants who were receiving HAART as opposed to 43.3% in those who were not (P<0.05). Conclusions HIV infected women who are eligible should start antiretroviral treatment prior to a pregnancy, in order to improve their immunological status. This measure associated to cotrimoxazole prophylaxis and ART could improve their survival. PMID:27186521

  5. Field Evaluation of a Semiautomated Method for Rapid and Simple Analysis of Recreational Water Microbiological Quality

    PubMed Central

    Anglès d'Auriac, Marc B.; Roberts, Hildegarde; Shaw, Terri; Sirevåg, Reidun; Hermansen, Leonila Fajardo; Berg, James D.

    2000-01-01

    An early warning system using a rapid enzymatic semiautomated method suitable for fecal coliform detection in recreational waters within 8 h was developed further and evaluated in this study. This rapid method was compared to the standard method followed in the United Kingdom. We used 1,011 samples originating from 206 different locations in Wales. When we assessed the presence or absence of fecal coliforms, targeting very low levels of contamination, we obtained 83.9% agreement between the rapid method and the lauryl sulfate broth-membrane filtration technique, whereas direct confirmation of the samples processed by the rapid method showed 89.3% agreement. Environmental enzymatic background activity was found to be the main limiting factor for this method. Owing to a specific and integrated handling of the results by the software of the instrument, the percentage of false-positive results (a consequence of enzymatic background) was successfully limited to 2.9% by the direct confirmation evaluation. However, 7.8% false-negative results due to “late-growers” had to be accepted in order to produce results within a working day. At present, the method can be used in a more conservative way to assess the environmental threshold of 100 CFU of fecal coliforms per 100 ml in recreational waters. The implications of our findings with regard to the applicability of rapid enzymatic methods are discussed. PMID:11010890

  6. Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered of Zucchini yellow mosaic virus.

    PubMed

    Kang, Minji; Seo, Jang-Kyun; Choi, Hoseong; Choi, Hong-Soo; Kim, Kook-Hyung

    2016-02-01

    The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits. PMID:26889118

  7. Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered of Zucchini yellow mosaic virus

    PubMed Central

    Kang, Minji; Seo, Jang-Kyun; Choi, Hoseong; Choi, Hong-Soo; Kim, Kook-Hyung

    2016-01-01

    The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits. PMID:26889118

  8. Simple & Rapid Generation of Complex DNA Profiles for the Undergraduate Laboratory

    ERIC Educational Resources Information Center

    Kass, David H.

    2007-01-01

    Deoxyribonucleic acid (DNA) profiles can be generated by a variety of techniques incorporating different types of DNA markers. Simple methods are commonly utilized in the undergraduate laboratory, but with certain drawbacks. In this article, the author presents an advancement of the "Alu" dimorphism technique involving two tetraplex polymerase…

  9. SIMPLE DEVICE TO DELIVER BEADS TO 96-WELL PLATES FOR RAPID RESUSPENSION OF CELLS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genomic sequencing involves sequence determination of inserts from a large numbers of plasmids of a genomic library. In high throughput DNA sequencing projects, cultures are grown in a 96-deep well plate format, centrifuged, and resuspended. Resuspension of cells was aided by use of a simple devic...

  10. Simple and rapid CD4 testing based on large-field imaging system composed of microcavity array and two-dimensional photosensor.

    PubMed

    Saeki, Tatsuya; Sugamura, Yuriko; Hosokawa, Masahito; Yoshino, Tomoko; Lim, Tae-Kyu; Harada, Manabu; Matsunaga, Tadashi; Tanaka, Tsuyoshi

    2015-05-15

    This study presents a novel method for CD4 testing based on one-shot large-field imaging. The large-field imaging system was fabricated by a microcavity array and a two-dimensional (2D) photosensor within the desk-top-sized instrument. The microcavity array was employed to separate leukocytes from whole blood based on differences in the size of leukocytes and other blood cells. The large-field imaging system with lower side irradiation enabled acquisition of cell signatures with high signal-to-noise ratio, because the metallic substrate of the microcavity array obstructed excessive excitation light. In this setting, dual-color imaging of CD4(+) and CD8(+) T cells was achieved within the entire image area (64 mm(2)) in 2s. The practical performance of the large-field imaging system was demonstrated by determining the CD4/CD8 ratio in a few microliter of control whole blood as small as those obtained by a finger prick. The CD4/CD8 ratios measured using the large-field imaging system correlated well with those measured by microscopic analysis. These results indicate that our proposed system provides a simple and rapid CD4 testing for the application of HIV/AIDS treatment. PMID:25192872

  11. Early diagnosis and retention in care of HIV-infected patients through rapid salivary testing: a test-and-treat fast track pilot study.

    PubMed

    Parisi, Maria Rita; Soldini, Laura; Negri, Silvia; Vidoni, Gian Marino; Gianotti, Nicola; Nozza, Silvia; Schlusnus, Karin; Dorigatti, Fernanda; Lazzarin, Adriano

    2016-01-01

    Aim of this study was to evaluate the efficacy and the retention-in-care of individuals diagnosed during six years of salivary HIV testing (EASY-test project). Among those linked-to-care at the Infectious Diseases Department of San Raffaele Hospital (Milan, Italy), the proportion of patients engaged, retained in care and virologically suppressed after the antiretroviral treatment was 96%, 100% and 95.2%, respectively. Results from our study suggest that salivary HIV testing may help bring to light cases of HIV infection otherwise undiagnosed, and thus favour a more rapid and wider reduction of the HIV infection burden at the population level. PMID:26922986

  12. HIV Rapid Testing in Substance Abuse Treatment: Implementation Following a Clinical Trial

    ERIC Educational Resources Information Center

    Haynes, L. F.; Korte, J. E.; Holmes, B. E.; Gooden, L.; Matheson, T.; Feaster, D. J.; Leff, J. A.; Wilson, L.; Metsch, L. R.; Schackman, B. R.

    2011-01-01

    The Substance Abuse Mental Health Services Administration has promoted HIV testing and counseling as an evidence-based practice. Nevertheless, adoption of HIV testing in substance abuse treatment programs has been slow. This article describes the experience of a substance abuse treatment agency where, following participation in a clinical trial,…

  13. A simple, rapid, inexpensive assay for toxic chemicals using a bacterial indicator

    SciTech Connect

    Botsford, J.L.; Hillaker, T.; Robertson, B.; Gonzales, M.; Benavidez, M.; Jones, B.; Baker, R.; Steen, W.; Pacheco, F.; Homer, V.; Lucero, O.; Matthews, M.; Koehler, V.

    1996-12-31

    A simple test for toxic chemicals has been developed. Rhizobium meliloti is combined with the toxic chemical. A tetrazolium dye, MTT (3-[4,5-Dimethylthiazol-2-yl]2,5-diphenyl-tetrazolium bromide) is added. The bacterium reduces this dye, causing the optical absorbance to increase dramatically. The increase can be determined with a simple spectrophotometer. Toxic chemicals and minerals inhibit the reduction of the dye. Presumably the dye serves as a terminal electron acceptor for electron transport. Toxic substances presumably damage the electron transport system. The results compare favorably with published results of tests using the Microtox{trademark} assay and with the Polytox{trademark} assay. This assay is simpler and requires no specialized equipment. It should be possible to use this assay in a third world situation.

  14. Introducing rapid oral–fluid HIV testing among high risk populations in Shandong, China: feasibility and challenges

    PubMed Central

    2014-01-01

    Background This study was conducted to ascertain the feasibility of using rapid oral fluid testing as an alternative HIV testing method in China. Method This is a mixed-method study among men who have sex with men (MSM), female sex workers (FSW) and VCT clients, conducted in 4 cities in Shandong Province. A pre-tested questionnaire was administered to 1137 participants through face-to-face interview to assess demographic characteristics, HIV testing histories and willingness to accept rapid oral fluid testing. VCT clients were provided with the saliva test kits for a screening test and errors in operation were recorded. Testing results were compared between oral and blood testing. Short feedback questionnaire was administered to 200 FSW who had undergone oral testing. Results The rate of willingness to take oral-fluid HIV testing among MSM, FSW and VCT clients was 72.8%, 72.1% and 67.4% respectively. Common errors recorded during test kit operation by the 229 VCT clients included: unpreparedness, wrong swab sampling, wrong dilution, wrong testing and inability to read test results. Advantages of oral testing listed by participants included: less intrusive, painlessness, easy self- testing and privacy. Disadvantages included perceived unreliable results (55.5%) and not nationally recognised (9%). Comparison of saliva and the blood testing results recorded a consistency rate of 0.970 (χ2 = 153.348, P < 0.001), implying an excellent consistency. Conclusion Introduction of oral rapid fluid testing as an alternative HIV testing method in China is highly feasible but with some challenges including low recognition and operation errors. PMID:24884431

  15. Rapid bacterial antibiotic susceptibility test based on simple surface-enhanced Raman spectroscopic biomarkers

    PubMed Central

    Liu, Chia-Ying; Han, Yin-Yi; Shih, Po-Han; Lian, Wei-Nan; Wang, Huai-Hsien; Lin, Chi-Hung; Hsueh, Po-Ren; Wang, Juen-Kai; Wang, Yuh-Lin

    2016-01-01

    Rapid bacterial antibiotic susceptibility test (AST) and minimum inhibitory concentration (MIC) measurement are important to help reduce the widespread misuse of antibiotics and alleviate the growing drug-resistance problem. We discovered that, when a susceptible strain of Staphylococcus aureus or Escherichia coli is exposed to an antibiotic, the intensity of specific biomarkers in its surface-enhanced Raman scattering (SERS) spectra drops evidently in two hours. The discovery has been exploited for rapid AST and MIC determination of methicillin-susceptible S. aureus and wild-type E. coli as well as clinical isolates. The results obtained by this SERS-AST method were consistent with that by the standard incubation-based method, indicating its high potential to supplement or replace existing time-consuming methods and help mitigate the challenge of drug resistance in clinical microbiology. PMID:26997474

  16. A simple, rapid bioassay for detecting effects of pollutants on bacteria

    SciTech Connect

    Bauer, N.J.; Seidler, R.J.; Knittel, M.D.

    1981-12-01

    A screening bioassay needs to be rapid, and sensitive. The bioassay is described which is accurate, inexpensive, and which utilizes bacteria as the toxicity predictor. The basis of the test involves measuring the kinetics of dissolved oxygen depletion by a mixed microbial population following exposure to a pollutant and allows results to be obtained in as little as 40 min. Pollutants tested were cadmium, copper, nickel, sulfate, diuron, pentachlorophenol, atrazine, tricholoracetic acid, dimethylformamide, and diazinon. (JMT)

  17. Rapid HIV-1 Disease Progression in Individuals Infected with a Virus Adapted to Its Host Population

    PubMed Central

    Shimizu, Akihisa; Zhu, Dayong; Han, Chungyong; Nakamura, Hitomi; Koga, Michiko; Kikuchi, Tadashi; Adachi, Eisuke; Koibuchi, Tomohiko; Gao, George F.; Brumme, Zabrina L.; Iwamoto, Aikichi

    2016-01-01

    HIV-1 escape from CTL is predictable based on the Human Leukocyte Antigen (HLA) class I alleles expressed by the host. As such, HIV-1 sequences circulating in a population of hosts will harbor escape mutations specific to the HLA alleles of that population. In theory, this should increase the frequency of escape mutation transmission to persons expressing the restricting HLA allele, thereby compromising host immunity to the incoming HIV-1 strain. However, the clinical impact of infection with HIV-1 containing immune escape mutations has not conclusively been demonstrated. Japan’s population features limited HLA diversity which is driving population-level HIV adaptation: for example, >60% of Japanese express HLA-A*24:02 and its associated Nef-Y135F escape mutation represents the population consensus. As such, Japan is an ideal population in which to examine this phenomenon. Here, we combine genetic and immunological analyses to identify A*24:02-positive individuals likely to have been infected with Y135F-containing HIV-1. Over a ~5 year follow-up, these individuals exhibited significantly lower CD4 counts compared to individuals inferred to have been infected with wild-type HIV-1. Our results support a significant negative clinical impact of pathogen adaptation to host pressures at the population level. PMID:26953793

  18. Rapid detection of HIV-1 proviral DNA for early infant diagnosis using recombinase polymerase amplification.

    PubMed

    Boyle, David S; Lehman, Dara A; Lillis, Lorraine; Peterson, Dylan; Singhal, Mitra; Armes, Niall; Parker, Mathew; Piepenburg, Olaf; Overbaugh, Julie

    2013-01-01

    Early diagnosis and treatment of human immunodeficiency virus type 1 (HIV-1) infection in infants can greatly reduce mortality rates. However, current infant HIV-1 diagnostics cannot reliably be performed at the point of care, often delaying treatment and compromising its efficacy. Recombinase polymerase amplification (RPA) is a novel technology that is ideal for an HIV-1 diagnostic, as it amplifies target DNA in <20 min at a constant temperature, without the need for complex thermocycling equipment. Here we tested 63 HIV-1-specific primer and probe combinations and identified two RPA assays that target distinct regions of the HIV-1 genome (long terminal repeat [LTR] and pol) and can reliably detect 3 copies of proviral DNA by the use of fluorescence detection and lateral-flow strip detection. These pol and LTR primers amplified 98.6% and 93%, respectively, of the diverse HIV-1 variants tested. This is the first example of an isothermal assay that consistently detects all of the major HIV-1 global subtypes. PMID:23549916

  19. Rapid Detection of HIV-1 Proviral DNA for Early Infant Diagnosis Using Recombinase Polymerase Amplification

    PubMed Central

    Boyle, David S.; Lehman, Dara A.; Lillis, Lorraine; Peterson, Dylan; Singhal, Mitra; Armes, Niall; Parker, Mathew; Piepenburg, Olaf; Overbaugh, Julie

    2013-01-01

    ABSTRACT Early diagnosis and treatment of human immunodeficiency virus type 1 (HIV-1) infection in infants can greatly reduce mortality rates. However, current infant HIV-1 diagnostics cannot reliably be performed at the point of care, often delaying treatment and compromising its efficacy. Recombinase polymerase amplification (RPA) is a novel technology that is ideal for an HIV-1 diagnostic, as it amplifies target DNA in <20 min at a constant temperature, without the need for complex thermocycling equipment. Here we tested 63 HIV-1-specific primer and probe combinations and identified two RPA assays that target distinct regions of the HIV-1 genome (long terminal repeat [LTR] and pol) and can reliably detect 3 copies of proviral DNA by the use of fluorescence detection and lateral-flow strip detection. These pol and LTR primers amplified 98.6% and 93%, respectively, of the diverse HIV-1 variants tested. This is the first example of an isothermal assay that consistently detects all of the major HIV-1 global subtypes. PMID:23549916

  20. Simple, specific, sensitive and rapid loop-mediated method for detecting Yersinia enterocolitica.

    PubMed

    Xu, Yun-Ming; Liu, Xi-Lin; Ma, Jing; Li, Yan-Song; Hu, Pan; Zou, De-Ying; Guo, Xing; Chen, Xiao-Feng; Tang, Feng; Liu, Nan-Nan; Wei, Li-Bin; Zhou, Yu; Liu, Zeng-Shan; Ren, Hong-Lin; Lu, Shi-Ying

    2014-05-01

    Yersinia enterocolitica (YE) is a main pathogenic bacterium causing diarrhea and yersiniosis occurs in both developed and developing countries with high incidence. YE in contaminated food is able to survive for a long duration even under cold storage, thereby enhancing the risk of food infection. In this study, a new loop-mediated isothermal amplification (LAMP) method showing the characteristics of simplicity, rapidity, high specificity and sensitivity was established by targeting outL of pathogenic YE. Two inner-primers and outer-primers were designed and LAMP reaction was optimized for Mg2+, betaine, dNTPs and inner primers concentrations, reaction temperature and time. Sensitivity and specificity of the LAMP assay was evaluated using YE genomic DNA and those of 44 different bacteria strains, respectively. Validation of LAMP detection method was by employing meat samples spiked with varying CFU of YE. The optimized LAMP assay was specific, capable of detecting 97 fg of genomic DNA (equivalent to 37 genome copies) of YE (100-fold more sensitive than PCR) and 80 CFU/ml of YE-spiked meat samples based on ethidium bromide stained amplicon bands on agarose gel-electrophoresis and on GelRed fluorescence of the LAMP reaction solution, respectively. This rapid, sensitive and specific LAMP technique should enable application in field inspection of Y. enterocolitica in food. PMID:24974652

  1. A simple hydrologic model for rapid prediction of runoff from ungauged coastal catchments

    NASA Astrophysics Data System (ADS)

    Wan, Yongshan; Konyha, Kenneth

    2015-09-01

    We developed a lumped conceptual rainfall-runoff model for rapid prediction of runoff generated in the unique hydrological setting with flat terrain, sandy soils, high groundwater table, and a dense drainage canal network in south Florida. The model is conceptualized as rainfall and evapotranspiration filling and emptying the root zone and excess rainfall recharging three storage zones. Outflows from these storage zones, routed with parallel arrangement of three linear reservoirs, represent different flow components of catchment runoff, i.e., slow drainage (shallow subsurface flow), medium drainage (interflow and saturation excess overland flow), and fast drainage (direct runoff from impervious urban areas or from water table management in agricultural land). The model is parsimonious with eight model parameters along with two optional water management parameters. A regionalization study was conducted through model parameterization to achieve target hydrological behavior of typical land uses, which are the most significant basin descriptor affecting catchment hydrology in south Florida. Cross validation with 16 gauged basins dominated by urban, agricultural, and natural lands, respectively, indicated that the model provides an effective tool for rapid prediction of runoff in ungauged basins using the regionalized model parameters. A case study is presented, involving application of the model to support real-time adaptive management to hydrological operations for protection of estuarine ecosystems.

  2. Rapid inflammasome activation in microglia contributes to brain disease in HIV/AIDS

    PubMed Central

    2014-01-01

    Background Human immunodeficiency virus type 1(HIV-1) infects and activates innate immune cells in the brain resulting in inflammation and neuronal death with accompanying neurological deficits. Induction of inflammasomes causes cleavage and release of IL-1β and IL-18, representing pathogenic processes that underlie inflammatory diseases although their contribution HIV-associated brain disease is unknown. Results Investigation of inflammasome-associated genes revealed that IL-1β, IL-18 and caspase-1 were induced in brains of HIV-infected persons and detected in brain microglial cells. HIV-1 infection induced pro-IL-1β in human microglia at 4 hr post-infection with peak IL-1β release at 24 hr, which was accompanied by intracellular ASC translocation and caspase-1 activation. HIV-dependent release of IL-1β from a human macrophage cell line, THP-1, was inhibited by NLRP3 deficiency and high extracellular [K+]. Exposure of microglia to HIV-1 gp120 caused IL-1β production and similarly, HIV-1 envelope pseudotyped viral particles induced IL-1β release, unlike VSV-G pseudotyped particles. Infection of cultured feline macrophages by the related lentivirus, feline immunodeficiency virus (FIV), also resulted in the prompt induction of IL-1β. In vivo FIV infection activated multiple inflammasome-associated genes in microglia, which was accompanied by neuronal loss in cerebral cortex and neurological deficits. Multivariate analyses of data from FIV-infected and uninfected animals disclosed that IL-1β, NLRP3 and caspase-1 expression in cerebral cortex represented key molecular determinants of neurological deficits. Conclusions NLRP3 inflammasome activation was an early and integral aspect of lentivirus infection of microglia, which was associated with lentivirus-induced brain disease. Inflammasome activation in the brain might represent a potential target for therapeutic interventions in HIV/AIDS. PMID:24886384

  3. Rapid quantification of microalgal lipids in aqueous medium by a simple colorimetric method.

    PubMed

    Mishra, Sanjiv K; Suh, William I; Farooq, Wasif; Moon, Myounghoon; Shrivastav, Anupama; Park, Min S; Yang, Ji-Won

    2014-03-01

    Identification of novel microalgal strains with high lipid productivity is one of the most important research topics in renewable biofuel research. However, the major bottleneck in the strain screening process is that currently known methods for the estimation of microalgal lipid are laborious and time-consuming. The present study successfully employed sulpho-phospho-vanillin (SPV) colorimetric method for direct quantitative measurement of lipids within liquid microalgal culture. The SPV reacts with lipids to produce a distinct pink color, and its intensity can be quantified using spectrophotometric methods by measuring absorbance at 530nm. This method was employed for a rapid quantification of intracellular lipid contents within Chlorella sp., Monoraphidium sp., Ettlia sp. and Nannochloropsis sp., all of which were found to have lipid contents ranging in between 10% and 30%. Subsequent analysis of the biomass using gas chromatography confirmed that our protocol is highly accurate (R(2)=0.99). PMID:24463407

  4. Rapid and simple purification of lysozyme from the egg shell membrane.

    PubMed

    Kozuka, Miyuki; Murao, Sato; Yamane, Takuya; Inoue, Tsutomu; Ohkubo, Iwao; Ariga, Hiroyoshi

    2015-01-01

    Lysozyme (EC 3.2.1.17) is a hydrolytic enzyme that cleaves the β-(1,4)-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in peptidoglycan, a major bacterial cell wall polymer. In the food industry, lysozyme is used as an additive mainly in the production of wine and beer. Lysozyme was found to be localized in the egg shell membrane. In this study, we found that lysozyme was easily purified from the egg shell membrane and that the enzyme also had antibacterial activity. Furthermore, we found that the antibacterial activity of purified lysozyme from the egg shell membrane was lower than that of purified lysozyme from the egg white at alkaline pH. The method for rapid purification of lysozyme developed in this study should contribute to the food industry. PMID:25994146

  5. Rapid Plateau border size variations expected in three simple experiments on 2D liquid foams.

    PubMed

    Gay, C; Rognon, P; Reinelt, D; Molino, F

    2011-01-01

    Up to a global scaling, the geometry of foams squeezed between two solid plates (2D GG foams) essentially depends on two independent parameters: the liquid volume fraction and the degree of squeezing (bubble thickness to diameter ratio). We describe it in two main asymptotic regimes: fully dry floor tiles, where the Plateau border radius is smaller than the distance between the solid plates, and dry pancakes, where it is larger. We predict a rapid variation of the Plateau border radius in one part of the pancake regime, namely when the Plateau border radius is larger than the inter-plate distance but smaller than the geometric mean of that distance and the bubble perimeter. This rapid variation is not related to any topological change in the foam: in all the regimes we consider, the bubbles remain in mutual lateral contact through films located at mid-height between both plates. We provide asymptotic predictions in different types of experiments on such 2D GG foams: when foam is being progressively dried or wetted, when it is being squeezed further or stretched, when it coarsens through film breakage or through inter-bubble gas diffusion. Our analysis is restricted to configurations close to equilibrium, as we do not include stresses resulting from bulk viscous flow or from non-homogeneous surfactant concentrations. We also assume that the inter-plate distance is sufficiently small for gravity to be negligible. The present work does not provide a method for measuring small Plateau border radii experimentally, but it indicates that large (and easily observable) Plateau borders should appear or disappear rather suddenly in some types of experiments with small inter-plate gaps. It also gives expected orders of magnitude that should be helpful for designing experiments on 2D GG foams. PMID:21253804

  6. Rapid and simple determination of T1 relaxation times in time-domain NMR by Continuous Wave Free Precession sequence.

    PubMed

    Moraes, Tiago Bueno; Monaretto, Tatiana; Colnago, Luiz Alberto

    2016-09-01

    Longitudinal (T1) and transverse (T2) relaxation times have been widely used in time-domain NMR (TD-NMR) to determine several physicochemical properties of petroleum, polymers, and food products. The measurement of T2 through the CPMG pulse sequence has been used in most of these applications because it denotes a rapid, robust method. On the other hand, T1 has been occasionally used in TD-NMR due to the long measurement time required to collect multiple points along the T1 relaxation curve. Recently, several rapid methods to measure T1 have been proposed. Those methods based upon single shot, known as Continuous Wave Free Precession (CWFP) pulse sequences, have been employed in the simultaneous measurement of T1 and T2 in a rapid fashion. However, these sequences can be used exclusively in instrument featuring short dead time because the magnitude of the signal at thermal equilibrium is required. In this paper, we demonstrate that a special CWFP sequence with a low flip angle can be a simple and rapid method to measure T1 regardless of instruments dead time. Experimental results confirmed that the method called CWFP-T1 may be used to measure both single T1 value and T1 distribution in heterogeneous samples. Therefore, CWFP-T1 sequence can be a feasible alternative to CPMG in the determination of physicochemical properties, particularly in processes where fast protocols are requested such as industrial applications. PMID:27376553

  7. Rapid and simple determination of T1 relaxation times in time-domain NMR by Continuous Wave Free Precession sequence

    NASA Astrophysics Data System (ADS)

    Moraes, Tiago Bueno; Monaretto, Tatiana; Colnago, Luiz Alberto

    2016-09-01

    Longitudinal (T1) and transverse (T2) relaxation times have been widely used in time-domain NMR (TD-NMR) to determine several physicochemical properties of petroleum, polymers, and food products. The measurement of T2 through the CPMG pulse sequence has been used in most of these applications because it denotes a rapid, robust method. On the other hand, T1 has been occasionally used in TD-NMR due to the long measurement time required to collect multiple points along the T1 relaxation curve. Recently, several rapid methods to measure T1 have been proposed. Those methods based upon single shot, known as Continuous Wave Free Precession (CWFP) pulse sequences, have been employed in the simultaneous measurement of T1 and T2 in a rapid fashion. However, these sequences can be used exclusively in instrument featuring short dead time because the magnitude of the signal at thermal equilibrium is required. In this paper, we demonstrate that a special CWFP sequence with a low flip angle can be a simple and rapid method to measure T1 regardless of instruments dead time. Experimental results confirmed that the method called CWFP-T1 may be used to measure both single T1 value and T1 distribution in heterogeneous samples. Therefore, CWFP-T1 sequence can be a feasible alternative to CPMG in the determination of physicochemical properties, particularly in processes where fast protocols are requested such as industrial applications.

  8. Rapid assessment of drug use and sexual HIV risk patterns among vulnerable drug-using populations in Cape Town, Durban and Pretoria, South Africa.

    PubMed

    Parry, Charles; Petersen, Petal; Carney, Tara; Dewing, Sarah; Needle, Richard

    2008-09-01

    This exploratory study examines the links between drug use and high-risk sexual practices and HIV in vulnerable drug-using populations in South Africa, including commercial sex workers (CSWs), men who have sex with men (MSM), injecting drug users (IDUs) and non-injecting drug users who are not CSWs or MSM (NIDUs). A rapid assessment ethnographic study was undertaken using observation, mapping, key informant interviews and focus groups in known 'hotspots' for drug use and sexual risk in Cape Town, Durban and Pretoria. Key informant (KI) and focus group interviews involved drug users and service providers. Purposeful snowball sampling and street intercepts were used to recruit drug users. Outcome measures included drug-related sexual HIV risk behaviour, and risk behaviour related to injection drug use, as well as issues related to service use. HIV testing of drug-using KIs was conducted using the SmartCheck Rapid HIV-1 Antibody Test. Non-injection drug use (mainly cannabis, methaqualone, crack cocaine and crystal methamphetamine) and injection drug use (mainly heroin) was occurring in these cities. Drug users report selling sex for money to buy drugs, and CSWs used drugs before, during and after sex. Most (70%) of the drug-using KIs offered HIV testing accepted and 28% were positive, with rates highest among CSWs and MSM. IDUs reported engaging in needle sharing and needle disposal practices that put them and others at risk for contracting HIV. There was a widespread lack of awareness about where to access HIV treatment and preventive services, and numerous barriers to accessing appropriate HIV and drug-intervention services were reported. Multiple risk behaviours of vulnerable populations and lack of access to HIV prevention services could accelerate the diffusion of HIV. Targeted interventions could play an important role in limiting the spread of HIV in and through these under-reached and vulnerable populations. PMID:18979044

  9. Rapid and Slow Progressors Show Increased IL-6 and IL-10 Levels in the Pre-AIDS Stage of HIV Infection

    PubMed Central

    de Medeiros, Rúbia M.; Valverde-Villegas, Jacqueline M.; Junqueira, Dennis M.; Gräf, Tiago; Lindenau, Juliana D.; de Mello, Marineide G.; Vianna, Priscila; Almeida, Sabrina E. M.; Chies, Jose Artur B.

    2016-01-01

    Cytokines are intrinsically related to disease progression in HIV infection. We evaluated the plasma levels of Th1/Th2/Th17 cytokines in extreme progressors, including slow (SPs) and rapid (RPs) progressors, who were thus classified based on clinical and laboratory follow-up covering a period of time before the initiation of HAART, ranging from 93–136.5 months for SPs and 7.5–16.5 months for RPs. Analyses were also performed based on the different stages of HIV infection (chronic, pre-HAART individuals—subjects sampled before initiating HAART but who initiated therapy from 12 to 24 months—and those receiving HAART). The plasma cytokine levels of 16 HIV-infected rapid progressors and 25 slow progressors were measured using a Human Th1/Th2/Th17 CBA kit. The IL-6 and IL-10 plasma levels differed significantly between the stages of HIV infection. The IL-6 levels were higher in slow progressors pre-HAART than in chronically infected SPs and HIV-seronegative individuals. The IL-10 levels were higher in slow progressors pre-HAART than in slow progressors receiving HAART and HIV-seronegative controls, and in rapid progressors, the IL-10 levels were higher in pre-HAART subjects than in HIV-seronegative controls. The results reflect the changes in the cytokine profile occurring during different clinical stages in HIV+ subjects. Our results suggest an association between increased IL-6 and IL-10 levels and pre-HAART stages independent of the slow or rapid progression status of the subjects. Thus, increased IL-6 and IL-10 levels could indicate a global inflammatory status and could be used as markers of the disease course in HIV-infected individuals. PMID:27214135

  10. Rapid and Slow Progressors Show Increased IL-6 and IL-10 Levels in the Pre-AIDS Stage of HIV Infection.

    PubMed

    de Medeiros, Rúbia M; Valverde-Villegas, Jacqueline M; Junqueira, Dennis M; Gräf, Tiago; Lindenau, Juliana D; de Mello, Marineide G; Vianna, Priscila; Almeida, Sabrina E M; Chies, Jose Artur B

    2016-01-01

    Cytokines are intrinsically related to disease progression in HIV infection. We evaluated the plasma levels of Th1/Th2/Th17 cytokines in extreme progressors, including slow (SPs) and rapid (RPs) progressors, who were thus classified based on clinical and laboratory follow-up covering a period of time before the initiation of HAART, ranging from 93-136.5 months for SPs and 7.5-16.5 months for RPs. Analyses were also performed based on the different stages of HIV infection (chronic, pre-HAART individuals-subjects sampled before initiating HAART but who initiated therapy from 12 to 24 months-and those receiving HAART). The plasma cytokine levels of 16 HIV-infected rapid progressors and 25 slow progressors were measured using a Human Th1/Th2/Th17 CBA kit. The IL-6 and IL-10 plasma levels differed significantly between the stages of HIV infection. The IL-6 levels were higher in slow progressors pre-HAART than in chronically infected SPs and HIV-seronegative individuals. The IL-10 levels were higher in slow progressors pre-HAART than in slow progressors receiving HAART and HIV-seronegative controls, and in rapid progressors, the IL-10 levels were higher in pre-HAART subjects than in HIV-seronegative controls. The results reflect the changes in the cytokine profile occurring during different clinical stages in HIV+ subjects. Our results suggest an association between increased IL-6 and IL-10 levels and pre-HAART stages independent of the slow or rapid progression status of the subjects. Thus, increased IL-6 and IL-10 levels could indicate a global inflammatory status and could be used as markers of the disease course in HIV-infected individuals. PMID:27214135

  11. A simple improved desolvation method for the rapid preparation of albumin nanoparticles.

    PubMed

    Jahanban-Esfahlan, Ali; Dastmalchi, Siavoush; Davaran, Soodabeh

    2016-10-01

    The current study tried to establish a simple and fast method for the preparation of BSA and HSA nanoparticles, based on an improved desolvation procedure under the aspect of a controllable particle size around 100nm for drug delivery applications. The Procedure used for the nanoparticles preparation was simplified by using a designed apparatus for controlling the addition of ethanol and it was used instead of conventional tubing pump which enabled the preparation of nanoparticles under defined conditions. By using EDC as cross-linker instead of glutharaldehyde, the time of nanoparticles preparation procedure was reduced to 3h. Several factors of the preparation process, such as the volume of the albumin solution, desolvating agent volume, the amount of cross-linker, the presence of salts and protein concentration were evaluated. Nanoparticles with smaller size were obtained under experimental conditions without the presence of salts or the use of buffers, 250mg of protein/4ml water, 5mg cross-linker, the addition of 4 and 8ml ethanol by using the designed apparatus to the HSA and BSA solution, respectively. By using this improved method, BSA and HSA nanoparticles of the size around 100nm and polydispersity below 0.2 were obtained. PMID:27177461

  12. A rapid and simple determination of caffeine in teas, coffees and eight beverages.

    PubMed

    Sereshti, Hassan; Samadi, Soheila

    2014-09-01

    Caffeine was extracted and preconcentrated by the simple, fast and green method of dispersive liquid-liquid microextraction (DLLME) and analysed by gas chromatography-nitrogen phosphorus detection (GC-NPD). The influence of main parameters affecting the extraction efficiency investigated and optimised. Under the optimal conditions, the method was successfully applied to determination of caffeine in different real samples including five types of tea (green, black, white, oolong teas and tea bag), two kinds of coffee (Nescafe coffee and coffee), and eight beverages (regular Coca Cola, Coca Cola zero, regular Pepsi, Pepsi max, Sprite, 7up, Red Bull and Hype).The limit of detection (LOD) and limit of quantification (LOQ) were 0.02 and 0.05 μg mL(-1), respectively. Linear dynamic range (LDR) was 0.05-500 μg mL(-1) and determination coefficient (R(2)) was 0.9990. The relative standard deviation (RSD) was 3.2% (n=5, C=1 μg mL(-1)). PMID:24731307

  13. A simple and rapid method for high-resolution visualization of single-ion tracks

    SciTech Connect

    Omichi, Masaaki; Choi, Wookjin; Sakamaki, Daisuke; Seki, Shu; Tsukuda, Satoshi; Sugimoto, Masaki

    2014-11-15

    Prompt determination of spatial points of single-ion tracks plays a key role in high-energy particle induced-cancer therapy and gene/plant mutations. In this study, a simple method for the high-resolution visualization of single-ion tracks without etching was developed through the use of polyacrylic acid (PAA)-N, N’-methylene bisacrylamide (MBAAm) blend films. One of the steps of the proposed method includes exposure of the irradiated films to water vapor for several minutes. Water vapor was found to promote the cross-linking reaction of PAA and MBAAm to form a bulky cross-linked structure; the ion-track scars were detectable at a nanometer scale by atomic force microscopy. This study demonstrated that each scar is easily distinguishable, and the amount of generated radicals of the ion tracks can be estimated by measuring the height of the scars, even in highly dense ion tracks. This method is suitable for the visualization of the penumbra region in a single-ion track with a high spatial resolution of 50 nm, which is sufficiently small to confirm that a single ion hits a cell nucleus with a size ranging between 5 and 20 μm.

  14. Simple and rapid method for the differentiation of Lens culinaris Medik. from false lentil species.

    PubMed

    Piergiovanni, Angela R; Taranto, Giovanni

    2005-08-24

    The practice of using both common vetch (Vicia sativa L.) and single-flower vetch (Vicia articulata Hornem.) seeds as food or feed is encouraged by the very high resemblance of their seeds with those of small-seeded lentil cultivars. Among the Vicieae, antinutritional and toxic factors are particularly important, because many species, containing high levels of these compounds, are not safe. A simple and fast capillary electrophoresis (CE) method was proposed for the differentiation of lentil cultivars from false lentil species (i.e., single-flour vetch and common vetch). Proteins were extracted from defatted milled seeds with an alcoholic/saline solution. Extracts were separated in an uncoated fused-silica capillary with iminodiacetic acid (IDA) isolectric buffer containing 0.05% hydroxypropylmethylcellulose (HPMC) and 20% acetonitrile. The presented method is useful also for the detection of contamination of whole or split seeds of lentil by vetch species. With respect to alternative techniques, such as DNA-based markers or thin-layer chromatography (TLC), CE has the advantages of being less expensive, faster, and fully automated. PMID:16104771

  15. A Simple and Rapid Protocol for Measuring Neutral Lipids in Algal Cells Using Fluorescence

    PubMed Central

    Storms, Zachary J.; Cameron, Elliot; de la Hoz Siegler, Hector; McCaffrey, William C.

    2014-01-01

    Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal fermentation processes and screening for new oil-rich strains requires a fast and reliable protocol for determination of intracellular lipid content. Current practices rely largely on gravimetric methods to determine oil content, techniques developed decades ago that are time consuming and require large sample volumes. In this paper, Nile Red, a fluorescent dye that has been used to identify the presence of lipid bodies in numerous types of organisms, is incorporated into a simple, fast, and reliable protocol for measuring the neutral lipid content of Auxenochlorella protothecoides, a green alga. The method uses ethanol, a relatively mild solvent, to permeabilize the cell membrane before staining and a 96 well micro-plate to increase sample capacity during fluorescence intensity measurements. It has been designed with the specific application of monitoring bioprocess performance. Previously dried samples or live samples from a growing culture can be used in the assay. PMID:24961928

  16. A simple and rapid microplate assay for glycoprotein-processing glycosidases

    SciTech Connect

    Kang, M.S.; Zwolshen, J.H.; Harry, B.S.; Sunkara, P.S. )

    1989-08-15

    A simple and convenient microplate assay for glycosidases involved in the glycoprotein-processing reactions is described. The assay is based on specific binding of high-mannose-type oligosaccharide substrates to concanavalin A-Sepharose, while monosaccharides liberated by enzymatic hydrolysis do not bind to concanavalin A-Sepharose. By the use of radiolabeled substrates (( 3H)glucose for glucosidases and (3H)mannose for mannosidases), the radioactivity in the liberated monosaccharides can be determined as a measure of the enzymatic activity. This principle was employed earlier for developing assays for glycosidases previously reported. These authors have reported the separation of substrate from the product by concanavalin A-Sepharose column chromatography. This procedure is handicapped by the fact that it cannot be used for a large number of samples and is time consuming. We have simplified this procedure and adapted it to the use of a microplate (96-well plate). This would help in processing a large number of samples in a short time. In this report we show that the assay is comparable to the column assay previously reported. It is linear with time and enzyme concentration and shows expected kinetics with castanospermine, a known inhibitor of alpha-glucosidase I.

  17. A simple and rapid protocol for measuring neutral lipids in algal cells using fluorescence.

    PubMed

    Storms, Zachary J; Cameron, Elliot; de la Hoz Siegler, Hector; McCaffrey, William C

    2014-01-01

    Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal fermentation processes and screening for new oil-rich strains requires a fast and reliable protocol for determination of intracellular lipid content. Current practices rely largely on gravimetric methods to determine oil content, techniques developed decades ago that are time consuming and require large sample volumes. In this paper, Nile Red, a fluorescent dye that has been used to identify the presence of lipid bodies in numerous types of organisms, is incorporated into a simple, fast, and reliable protocol for measuring the neutral lipid content of Auxenochlorella protothecoides, a green alga. The method uses ethanol, a relatively mild solvent, to permeabilize the cell membrane before staining and a 96 well micro-plate to increase sample capacity during fluorescence intensity measurements. It has been designed with the specific application of monitoring bioprocess performance. Previously dried samples or live samples from a growing culture can be used in the assay. PMID:24961928

  18. A simple and rapid test of physical performance inchronic obstructive pulmonary disease.

    PubMed

    Albarrati, Ali Mufraih; Gale, Nichola S; Enright, Stephanie; Munnery, Margaret M; Cockcroft, John R; Shale, Dennis J

    2016-01-01

    Impaired physical performance is common in chronic obstructive pulmonary disease (COPD), but its assessment can be difficult in routine clinical practice. We compared the timed up and go (TUG) test and other easily applied assessments of physical performance with the 6-minute walk distance (6MWD). In a longitudinal study of comorbidities in COPD, submaximal physical performance was determined in 520 patients and 150 controls using the TUG test and 6MWD. Spirometry, body composition, handgrip strength, the COPD assessment test, St George's Respiratory Questionnaire (SGRQ), and the modified Medical Research Council dyspnoea scale were also determined. Patients and controls were similar in age, body mass index, and sex proportions. The TUG in the patients was greater than that in the control group, P=0.001, and was inversely related to 6MWD (r=-0.71, P<0.001) and forced expiratory volume in one second predicted (r=-0.19, P<0.01) and was directly related to the SGRQ activity (r=0.39, P<0.001), SGRQ total (r=0.37, P<0.001), and total COPD assessment test scores (r=0.37, P<0.001). The TUG identified the difference in physical performance between patients and controls. The TUG test and validated questionnaires provide a measure of physical performance, which is rapid and could be used in clinical practice. PMID:27536090

  19. A rapid and simple method for the separation of pure lymphocytes from horse blood.

    PubMed

    Zizzadoro, Claudia; Belloli, Chiara; Badino, Paola; Ormas, Paolo

    2002-10-01

    A method for the separation of pure and viable lymphocytes and granulocytes from the same blood sample in horses was reported. By centrifuging equine heparinized blood at 100 xg for 10 min at room temperature (r.t.), the resulting supernatant plasma was an almost pure (97.71 +/- 0.30%; n = 15) suspension of highly viable (98.72 +/- 0.28%) lymphocytes. When sodium citrate was used as an anticoagulant, lymphocyte suspensions collected in the same manner showed lower purity (87.89 +/- 1.59%; n = 9) and higher yields (56.56 +/- 3.89%, n = 9 versus 36.11 +/- 2.23%, n = 15). Where needed, a further centrifugation at 250 xg for 3 min (r.t.) of heparinized lymphocyte preparations removed an average of 87.39% (n = 15) contaminating platelets. A suspension of 85.96 +/- 2.20% pure granulocytes (93.23 +/- 1.74% neutrophils; n = 14) with minimal contamination by erythrocytes and high viability (93.11 +/- 1.26%) was obtained by performing a flash red blood cell lysis on the white-greyish layer resulting from the centrifugation of the heparinized blood samples. Among the several methods available, the procedure described herein is easy, rapid, cheap and reproducible. PMID:12208055

  20. Development and evaluation of a rapid and simple procedure for detection of Pneumocystis carinii by PCR.

    PubMed Central

    Cartwright, C P; Nelson, N A; Gill, V J

    1994-01-01

    We report the development of a simplified PCR-based assay for the detection of Pneumocystis carinii DNA in clinical specimens. The adoption of a rapid DNA extraction procedure and the introduction of a type of enzyme-linked immunosorbent assay for PCR product detection enabled this procedure to be carried out in a single working day in a clinical microbiology laboratory. The PCR assay was prospectively compared with an immunofluorescent-antibody (FA) staining method for the detection of P. carinii in induced sputum and bronchoalveolar lavage (BAL) specimens. The results of the study showed that, for induced sputum specimens, FA staining had a sensitivity of 78% (32 of 41 specimens) and a specificity of 100% (166 of 166 specimens); PCR was 100% (41 of 41 specimens) sensitive and 98% (162 of 166 specimens) specific. For BAL specimens, FA staining was 100% sensitive (21 of 21 specimens) and 100% specific (133 of 133 specimens), and PCR had a sensitivity of 100% (21 of 21 specimens) and a specificity of 99% (132 of 133 specimens). These results strongly suggest that use of our PCR-based assay could effect clinically useful improvements in the sensitivity of induced sputum specimens for the detection of P. carinii. Images PMID:7929749

  1. Rapid detection of neurotoxic insecticides in food using disposable acetyicholinesterase-biosensors and simple solvent extraction.

    PubMed

    Schulze, Holger; Schmid, Rolf D; Bachmann, Till T

    2002-01-01

    The extensive use of pesticides to protect agricultural crops necessitates reliable tools for the detection of residues in food and water, thus ensuring environmental protection and consumer safety. Neuroinhibitors such as organophosphates and carbamates in particular, represent a potential hazard to human health. These compounds are frequently found in food, but conventional methods of analysis are limited as they are either time consuming or not sufficiently sensitive. As a result, a rapid and sensitive biosensor test based on AChE-inhibition was developed. The disposable AChE-biosensor was directly applied in solvent extracts of food samples using isooctane as extraction solvent. A complete assay could be performed in less than 2 h. Recovery rates of 84% were obtained in tests with spiked orange juice samples. Tests in food samples with a lower water content resulted in reduced recovery rates (44% for peach pap baby food). Phosphorothionate insecticides in food could be detected after direct oxidation with N-bromosuccinimide and solvent extraction. The assay displayed a detection limit of 2 microg/kg paraoxon, which was sufficient for the monitoring of maximum residue limits in food according to EU regulations. PMID:11936097

  2. A simple and rapid test of physical performance in chronic obstructive pulmonary disease

    PubMed Central

    Albarrati, Ali Mufraih; Gale, Nichola S; Enright, Stephanie; Munnery, Margaret M; Cockcroft, John R; Shale, Dennis J

    2016-01-01

    Impaired physical performance is common in chronic obstructive pulmonary disease (COPD), but its assessment can be difficult in routine clinical practice. We compared the timed up and go (TUG) test and other easily applied assessments of physical performance with the 6-minute walk distance (6MWD). In a longitudinal study of comorbidities in COPD, submaximal physical performance was determined in 520 patients and 150 controls using the TUG test and 6MWD. Spirometry, body composition, handgrip strength, the COPD assessment test, St George’s Respiratory Questionnaire (SGRQ), and the modified Medical Research Council dyspnoea scale were also determined. Patients and controls were similar in age, body mass index, and sex proportions. The TUG in the patients was greater than that in the control group, P=0.001, and was inversely related to 6MWD (r=−0.71, P<0.001) and forced expiratory volume in one second predicted (r=−0.19, P<0.01) and was directly related to the SGRQ activity (r=0.39, P<0.001), SGRQ total (r=0.37, P<0.001), and total COPD assessment test scores (r=0.37, P<0.001). The TUG identified the difference in physical performance between patients and controls. The TUG test and validated questionnaires provide a measure of physical performance, which is rapid and could be used in clinical practice. PMID:27536090

  3. A simple, rapid and sensitive FRET assay for botulinum neurotoxin serotype B detection.

    PubMed

    Guo, Jiubiao; Xu, Ci; Li, Xuechen; Chen, Sheng

    2014-01-01

    Botulinum neurotoxins (BoNTs), the most potent naturally-occurring neurotoxins known to humans, comprise seven distinct serotypes (BoNT/A-G), each of which exhibits unique substrate specificity. Many methods have been developed for BoNT detection, in particular for BoNT/A, with various complexity and sensitivity, while substrate based FRET assay is considered as the most widely used approach due to its simplicity and sensitivity. In this study, we designed a vesicle-associated membrane protein 2 (VAMP2) based FRET assay based on the understanding of the VAMP2 and light chain/B (LC/B) interactions in our previous studies. The current design constituted the shortest peptide, VAMP2 (63-85), with FRET dyes (EDAN and Dabcyl) labelled at position 76 and 85, respectively, which showed minimal effect on VAMP2 substrate catalysis by LC/B and therefore enhanced the sensitivity of the assay. The FRET peptide, designated as FVP-B, was specific to LC/B, with a detection sensitivity as low as ∼20 pM in 2 h. Importantly, FVP-B showed the potential to be scaled up and used in high throughput screening of LC/B inhibitor. The currently developed FRET assay is one of the most economic and rapid FRET assays for LC/B detection. PMID:25437190

  4. A rapid and simple one-step F-18 labeling of peptides

    PubMed Central

    Jacobson, Orit; Zhu, Lei; Ma, Ying; Weiss, Ido D.; Sun, Xilin; Niu, Gang; Kiesewetter, Dale O.; Chen, Xiaoyuan

    2011-01-01

    Objectives Labeling biomolecules with 18F is usually done through coupling with prosthetic groups, which requires several time-consuming radiosynthesis steps and therefore results in low labeling yield. In this study we designed a simple one-step 18F-labeling strategy to replace the conventional complex and long process of multiple-step radiolabeling procedure. Methods Both Monomeric and dimeric cyclic RGD peptides were modified to contain 4-NO2-3-CF3 arene as precursors for direct 18F labeling. Binding of the two functionalized peptides to integrin αvβ3 was tested in vitro using MDA-MB-435 human breast cell line. The most promising functionalized peptide, the dimeric cyclic RGD, was further evaluated in vivo in an orthotopic MDA-MB-435 tumor xenograft model. Results The use of relatively low amount of precursor (~0.5μmol), gave reasonable yield, ranging from 7–23% (decay corrected, calculated from start of synthesis) after HPLC purification. Overall reaction time was 40 min and the specific activity of the labeled peptide was high. Modification of RGD peptides did not significantly change the biological binding affinities of the modified peptides. Small animal PET and biodistribution studies revealed integrin specific tumor uptake and favorable biokinetics. Conclusions We have developed a novel one-step 18F radiolabeling strategy for peptides that contain a specific arene group, which shortens reaction time and labor significantly, requires low amount of precursor and results in specific activity of 79 ± 13 GBq/μmol. Successful introduction of 4-fluoro-3-trifluoromethylbenzamide into RGD peptides may be a general strategy applicable to other biologically active peptides and proteins. PMID:21338096

  5. Evaluation of Rapidly Disintegrating Vaginal Tablets of Tenofovir, Emtricitabine and Their Combination for HIV-1 Prevention

    PubMed Central

    Clark, Meredith R.; Peet, M. Melissa; Davis, Sarah; Doncel, Gustavo F.; Friend, David R.

    2014-01-01

    Vaginal tablets are being developed as an alternative to gels as an inexpensive, discreet dosage form for the administration of microbicides. This work describes the pharmacokinetic (PK) evaluation of rapidly disintegrating vaginal tablets containing tenofovir (TFV, 10 mg), emtricitabine (FTC, 10 mg), and the combination of TFV and FTC (10 mg each) under in vitro and in vivo conditions, and in direct comparison to the clinical TFV 1% gel, a microbicide product in Phase III clinical testing. The PK of TFV and FTC from tablets were also evaluated in female rabbits following intravaginal administration. Direct comparison of a single dose of TFV tablets (intact or predissolved at 10 mg/mL) and TFV 1% gel showed no differences in the vaginal PK of TFV between groups; however systemic bioavailability of TFV was significantly higher from the gel. When rabbits were dosed either once or daily for seven days with intact tablets of TFV, FTC, or the combination of TFV/FTC, vaginal and systemic concentrations of TFV and FTC were unaffected by co-formulation. Moreover, plasma PK parameters were similar following a single dose or seven once-daily doses. Tissue concentrations of TFV and FTC in the cranial vagina 4 h after administration ranged between 104 and 105 ng/g. Concentrations of TFV-diphospate (TFV-DP, the active metabolite) were also high (over 103 ng/g or about 3000 to 6000 fmol/mg) in the cranial vagina 4 h after administration and similar to those measured following administration of TFV 1% gel. These data demonstrate that rapidly disintegrating vaginal tablets may be a suitable topical microbicide dosage form providing similar vaginal TFV PK to that of TFV 1% gel. The data also support co-administration of FTC with TFV in a single vaginal tablet to create a combination microbicide in a simple and inexpensive dosage form. PMID:25494201

  6. Rapid universal solublization and analysis of bacterial spores using a simple flow-through ultrahigh-temperature capillary device

    NASA Astrophysics Data System (ADS)

    West, Jay A.; Hukari, Kyle W.; Renzi, Ronald F.; Patel, Kamlesh

    2004-12-01

    Rapid identification of viral and bacterial species is dependent of the ability to manipulate biological agents into a form where they are directly analyzed. Many of these species of interest, such as bacterial spores, are inherently hearty and very difficult to lyse or solubilize. Standard protocols for spore inactivation include, chemical treatment, sonication, pressure and thermal lysis. While these protocols are effective for the inactivation of these agents they are less well suited for sample preparation for analysis using capillary electrophoresis techniques. In order to overcome this difficulty we fabricated a simple capillary device to perform thermal lysis of vegatative bacterial cells and bacterial spores. Using an ethylene glycol buffer to super heat bacterial spores we were able to perform rapid flow through lysis and solubilzation of these agents. This device was then coupled to a sample preparation station for on-line fluorescamine dye lableling and buffer exchange for direct analysis using a miniaturized capillary electrophoresis instrument. Using this integrated device were we enabled to perform sample lysis, labeling and protein fingerprint analysis of vegatative bacterial cells, bacterial spores and viruses in less than 10 minutes. The described device is simple, inexpensive and easily integratable with various microfluidic devices.

  7. An improved reverse dot hybridization for simple and rapid detection of adefovir dipivoxil-resistant hepatitis B virus.

    PubMed

    Hu, Y; Zhang, W L; Xie, S L; Zhao, Y; Hu, J L; Cai, X F; Lai, G Q; Huang, A L

    2012-01-01

    Early detection of adefovir dipivoxil-resistant mutants during long-term treatment of chronic hepatitis B virus (HBV) infection with this drug is of great clinical importance. We developed an improved reverse dot hybridization test for simple and rapid detection of the rtA181V/T and rtN236T mutations associated with adefovir dipivoxil resistance in chronic hepatitis B patients. Probes were designed for genotypes B, C, and D of this resistance characteristic; a total of 70 clinical samples were analyzed with this improved reverse dot hybridization assay. Its usefulness was validated by comparing with sequencing data. Discordant results were confirmed by subclone sequencing. This reverse dot hybridization assay was sufficiently sensitive to detect 10(3) copies/mL; it also detected adefovir dipivoxil-resistant mutant strains when they comprised more than 5% of a mixed virus population. This reverse dot hybridization array correctly identified adefovir dipivoxil-resistant mutants; it had high concordance (98.5%) with direct sequencing data. There was no clear relationship between the HBV genotype and the development of adefovir dipivoxil-resistant mutants. This reverse dot hybridization assay proved to be simple and rapid for detection of rtA181V/T and rtN236T mutations associated with resistance to adefovir dipivoxil. PMID:22290465

  8. HIV

    PubMed Central

    Chawla, Sumit; Sahoo, Soumya Swaroop; Jain, Rambilas; Khanna, Pardeep; Mehta, Bharti; Singh, Inderjeet

    2014-01-01

    Getting to zero: zero new HIV infections, zero deaths from AIDS-related illness, zero discrimination is the theme of World AIDS Day 2012. Given the spread of the epidemic today, getting to zero may sound difficult, but significant progress is underway. The total annual loss for the entire country due to HIV is 7% of GDP, which exceeds India’s annual health expenditure in 2004. The additional loss due to loss of labor income and increased medical expenditure as measured by the external transfers, account for 5% of the country’s health expenditure and 0.23% of GDP. Given that the HIV incidence rate is only 0.27% in India, these losses are quite staggering. Despite the remarkable achievements in development of anti-retroviral therapies against HIV and the recent advances in new prevention technologies, the rate of new HIV infections continue to outpace efforts on HIV prevention and control. Thus, the development of a safe and effective vaccine for prevention and control of AIDS remains a global public health priority and the greatest opportunity to eventually end the AIDS pandemic. PMID:24056755

  9. Simple, rapid, and reliable detection of Escherichia coli O26 using immunochromatography.

    PubMed

    Yonekita, Taro; Fujimura, Tatsuya; Morishita, Naoki; Matsumoto, Takashi; Morimatsu, Fumiki

    2013-05-01

    Shiga toxin-producing Escherichia coli (STEC) O26 has been increasingly associated with diarrheal disease all over the world. We developed an immunochromatographic (ic) strip for the rapid detection of E. coli O26 in food samples. To determine the specificity of the IC strip, pure cultures of 67 E. coli and 22 non-E. coli strains were tested with the IC strip. The IC strip could detect all (18 of 18) E. coli O26 strains tested and did not react with strains of any other E. coli serogroup or non-E. coli strains tested (0 of 71). The minimum detection limits for E. coli O26 were 2.2 × 10(3) to 1.0 × 10(5) cfu/ml. To evaluate the ability of the IC strip to detect E. coli O26 in food, 25-g food samples (ground beef, beef liver, ground chicken, alfalfa sprout, radish sprout, spinach, natural cheese, and apple juice) were spiked with E. coli O26. The IC strip was able to detect E. coli O26 at very low levels (approximately 1 cfu/25 g of food samples) after an 18-h enrichment, and the IC strip results were in 100% agreement with the results of the culture method and pcr assay. When 115 meat samples purchased from supermarkets were tested, 5 were positive for E. coli O26 with the IC strip; these results were confirmed with a pcr assay. These results suggest that the IC strip is a useful tool for detecting E. coli O26 in food samples. PMID:23643115

  10. Smart HIV testing system.

    PubMed

    El Kateeb, Ali; Law, Peter; Chan, King

    2005-06-01

    The quick HIV testing method called "MiraWell Rapid HIV Test" uses a specialized testing kit to determine whether an individual's blood is contaminated with the HIV virus or not. When a drop of blood is placed on the center of the testing kit, a simple pattern will appear in the middle of the kit to indicate the test status, i.e., positive or negative. This HIV test should be done in a small clinic or in a lab and the test must be conducted by a trained technician. A smart HIV testing system was developed through this research to eliminate the human error that is associated with the use of the quick HIV testing kits. Also, the smart HIV system will improve the testing productivity in comparison to those achieved by the trained technicians. In this research, we have developed a cost-effective system that analyzes the image produced by the HIV kits. We have used a System-On-Chip (SOC) design approach based on the Field Programmable Gate Array (FPGA) technology and the Xilinx Virtex SOC chip in building the system's prototype. The system used a CMOS digital camera to capture the image and an FPGA chip to process the captured image and send the testing results to the display unit. The system can be used in small clinics and pharmacies and eliminates the need for trained technicians. The system has been tested successfully and 98% of the tests were correct. PMID:16078623

  11. Rapid and simple detection of methicillin-resistance staphylococcus aureus by orfX loop-mediated isothermal amplification assay

    PubMed Central

    2014-01-01

    Background Methicillin-resistant Staphylococcus aureus (MRSA) has become one of the most prevalent pathogens responsible for nosocomial infections throughout the world. As clinical MRSA diagnosis is concerned, current diagnostic methodologies are restricted by significant drawbacks and novel methods are required for MRSA detection. This study aimed at developing a simple loop-mediated isothermal amplification (LAMP) assay targeting on orfX for the rapid detection of methicillin-resistance Staphylococcus aureus (MRSA). Results The protocol was designed by targeting orfX, a highly conserved open reading frame in S. aureus. One hundred and sixteen reference strains, including 52 Gram-positive and 64 Gram-negative isolates, were included for evaluation and optimization of the orfX-LAMP assay. This assay had been further performed on 667 Staphylococcus (566 MRSA, 25 MSSA, 53 MRCNS and 23 MSCNS) strains and were comparatively validated by PCR assay using primers F3 and B3, with rapid template DNA processing, simple equipments (water bath) and direct result determination (both naked eye and under UV light) applied. The indispensability of each primer had been confirmed, and the optimal amplification was obtained under 65°C for 45 min. The 25 μl reactant was found to be the most cost-efficient volume, and the detection limit was determined to be 10 DNA copies and 10 CFU/reaction. High specificity was observed when orfX-LAMP assay was subjected to 116 reference strains. For application, 557 (98.4%, 557/566) and 519 (91.7%, 519/566) tested strains had been detected positive by LAMP and PCR assays. The detection rate, positive predictive value (PPV) and negative predictive value (NPV) of orfX-LAMP were 98.4%, 100% and 92.7% respectively. Conclusions The established orfX-LAMP assay had been demonstrated to be a valid and rapid detection method on MRSA. PMID:24456841

  12. [Simple and rapid screening for psychotropic natural products using Direct Analysis in Real Time (DART)-TOFMS].

    PubMed

    Kawamura, Maiko; Kikura-Hanajiri, Ruri; Goda, Yukihiro

    2009-06-01

    Direct Analysis in Real Time (DART) is a novel ionization technique that provides for the rapid ionization of small molecules under ambient conditions. To investigate the trend of non-controlled psychotropic plants of abuse in Japan, a rapid screening method, without sample preparation, was developed using DART-time of flight mass spectrometer (TOFMS) for plant products. The major psychotropic constituents of these products were determined using liquid chromatography-mass spectrometry (LC/MS). As a result of the DART-TOFMS analyses of 36 products, the protonated molecular ions [M+H](+), corresponding to 6 kinds of major hallucinogenic constituents (mescaline, salvinorin A, N,N-dimethyltryptamine, harmine, harmaline and lysergamide), were detected in 21 products. It was possible to estimate their accurate elemental compositions through exact mass measurements. These results were consistent with those of the LC/MS analyses and the contents of the 6 psychotropic constituents were in the range from 0.05 to 45 microg/mg. Typical controlled narcotic drugs, tetrahydrocannabinol, opioid alkaloids and psilocin were also directly detected in marijuana cigarette, opium gum and magic mushroom respectively. Although it is difficult to estimate the matrix effects caused by other plant ingredients, the DART-TOFMS could be useful as a simple and rapid screening method for the targeted psychotropic natural products, because it provides the molecular information of the target compounds without time-consuming extraction and pre-treatment steps. PMID:19483414

  13. Initial Accuracy of HIV Rapid Test Kits Stored in Suboptimal Conditions and Validity of Delayed Reading of Oral Fluid Tests

    PubMed Central

    Choko, Augustine T.; Taegtmeyer, Miriam; MacPherson, Peter; Cocker, Derek; Khundi, McEwen; Thindwa, Deus; Sambakunsi, Rodrick S.; Kumwenda, Moses K.; Chiumya, Kondwani; Malema, Owen; Makombe, Simon D.; Webb, Emily L.; Corbett, Elizabeth L.

    2016-01-01

    Objectives To evaluate the effect of storing commonly used rapid diagnostic tests above manufacturer-recommended temperature (at 37°C), and the accuracy of delayed reading of oral fluid kits with relevance to HIV self-testing programmes. Design A quality assurance study of OraQuick (OraSure), Determine HIV 1/2™ (Alere) and Uni-Gold™ (Recombigen®). Methods Consecutive adults (≥18y) attending Ndirande Health Centre in urban Blantyre, Malawi in January to April 2012 underwent HIV testing with two of each of the three rapid diagnostic test kits stored for 28 days at either 18°C (optimally-stored) or at 37°C (pre-incubated). Used OraQuick test kits were stored in a laboratory for delayed day 1 and subsequent monthly re-reading was undertaken for one year. Results Of 378 individuals who underwent parallel testing, 5 (1.3%) were dropped from the final analysis due to discordant or missing reference standard results (optimally-stored Determine and Uni-Gold). Compared to the diagnostic reference standard, OraQuick had a sensitivity of 97.2% (95% CI: 93.6–99.6). There were 7 false negative results among all test kits stored at 37°C and three false negatives among optimally stored kits. Excellent agreement between pre-incubated tests and optimally-stored tests with Kappa values of 1.00 for Determine and Uni-Gold; and 0.97 (95% CI: 0.95; 1.00) for OraQuick were observed. There was high visual stability on re-reading of OraQuick, with only 1/375 pre-incubated and 1/371 optimally-stored OraQuick kits changing from the initial result over 12 months. Conclusion Erroneous results observed during HIV testing in low income settings are likely to be due to factors other than suboptimal storage conditions. Re-reading returned OraQuick kits may offer a convenient and accurate quality assurance approach, including in HIV self-testing programmes. PMID:27336161

  14. Combination Antiretroviral Therapy With Raltegravir Leads to Rapid Immunologic Reconstitution in Treatment-Naive Patients With Chronic HIV Infection

    PubMed Central

    Pallikkuth, Suresh; Fischl, Margaret A.; Pahwa, Savita

    2013-01-01

    Background. In treatment-naive, human immunodeficiency virus (HIV)–infected persons, combination antiretroviral therapy (cART) incorporating raltegravir (RAL) is highly effective for virologic suppression, but characteristics of immunologic recovery have not been described. Methods. We performed a 48-week substudy of 15 patients, median age 40 years, within a phase 2 randomized trial of RAL-cART in treatment-naive patients with chronic HIV infection. Results. Plasma viral load decreased from 5.2 ± 5.3 log10 HIV RNA copies/mL to 2.2 ± 2.4 log10 copies/mL at week 4, reaching <50 copies/mL at week 8 in 13 of 15 patients. Total CD4 T cells increased at week 4, as did central memory CD4 T cells in association with reduction of the immune activation markers HLA-DR and CD38 and immune exhaustion marker PD1 in CD4 and CD8 T cells. Naive CD4 T cells increased at week 24 with appearance of HIV gag–specific interleukin 2, interferon-γ, and CD107a responses in CD4 and CD8 T cells at week 48. Plasma lipopolysaccharide and soluble CD14 decreased, but at week 48 were elevated as compared to healthy volunteers. Altogether, the week 48 immune profile was more favorable in patients taking RAL-cART than in patients treated with non–RAL-cART. Conclusions. RAL in first-line treatment regimens results in rapid immune reconstitution with residual low-level microbial translocation. PMID:23922374

  15. A simple and rapid colloidal gold-based immunochromatogarpic strip test for detection of FMDV serotype A.

    PubMed

    Jiang, Tao; Liang, Zhong; Ren, Wei-wei; Chen, Juan; Zhi, Xiao-ying; Qi, Guang-yu; Liu, Xiang-tao; Cai, Xue-peng

    2011-02-01

    A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test, affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip, the capture antibody was laid on a sample pad, the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7 ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C, Swine vesicular disease (SVD), Vesicular stomatitis virus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically, the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple, easy and fast for clinical testing on field sites; no special instruments and skills are required, and the result can be obtained within 15 min. To our knowledge, this is the first rapid immunochromatogarpic assay for serotype A of FMDV. PMID:21331888

  16. Nucleocapsid Annealing-Mediated Electrophoresis (NAME) Assay Allows the Rapid Identification of HIV-1 Nucleocapsid Inhibitors

    PubMed Central

    Sosic, Alice; Cappellini, Marta; Scalabrin, Matteo; Gatto, Barbara

    2015-01-01

    RNA or DNA folded in stable tridimensional folding are interesting targets in the development of antitumor or antiviral drugs. In the case of HIV-1, viral proteins involved in the regulation of the virus activity recognize several nucleic acids. The nucleocapsid protein NCp7 (NC) is a key protein regulating several processes during virus replication. NC is in fact a chaperone destabilizing the secondary structures of RNA and DNA and facilitating their annealing. The inactivation of NC is a new approach and an interesting target for anti-HIV therapy. The Nucleocapsid Annealing-Mediated Electrophoresis (NAME) assay was developed to identify molecules able to inhibit the melting and annealing of RNA and DNA folded in thermodynamically stable tridimensional conformations, such as hairpin structures of TAR and cTAR elements of HIV, by the nucleocapsid protein of HIV-1. The new assay employs either the recombinant or the synthetic protein, and oligonucleotides without the need of their previous labeling. The analysis of the results is achieved by standard polyacrylamide gel electrophoresis (PAGE) followed by conventional nucleic acid staining. The protocol reported in this work describes how to perform the NAME assay with the full-length protein or its truncated version lacking the basic N-terminal domain, both competent as nucleic acids chaperones, and how to assess the inhibition of NC chaperone activity by a threading intercalator. Moreover, NAME can be performed in two different modes, useful to obtain indications on the putative mechanism of action of the identified NC inhibitors. PMID:25650789

  17. A Rapid and Simple LC-MS Method Using Collagen Marker Peptides for Identification of the Animal Source of Leather.

    PubMed

    Kumazawa, Yuki; Taga, Yuki; Iwai, Kenji; Koyama, Yoh-Ichi

    2016-08-01

    Identification of the animal source of leather is difficult using traditional methods, including microscopic observation and PCR. In the present study, a LC-MS method was developed for detecting interspecies differences in the amino acid sequence of type I collagen, which is a major component of leather, among six animals (cattle, horse, pig, sheep, goat, and deer). After a dechroming procedure and trypsin digestion, six tryptic peptides of type I collagen were monitored by LC-MS in multiple reaction monitoring mode for the animal source identification using the patterns of the presence or absence of the marker peptides. We analyzed commercial leathers from various production areas using this method, and found some leathers in which the commercial label disagreed with the identified animal source. Our method enabled rapid and simple leather certification and could be applied to other animals whether or not their collagen sequences are available in public databases. PMID:27397145

  18. Development and Validation of an Inexpensive, Simple, and Rapid Technique for Measuring the Accuracy of Extemporaneously Compounded Pharmaceuticals.

    PubMed

    Meek, Claudia; Rothardt, Andrew; Evans, Jason; Thurman, Rosanne; Ashworth, Lisa; Leff, Richard

    2015-01-01

    Use of ultraviolet detection to quantitate analytes is a basic concept of analytical chemistry. The basis of this application is well defined by Beer-Lambert's law. To this end, the authors applied Beer-Lambert's law as a simple and rapid tool to measure the accuracy of extemporaneously compounded pharmaceuticals. Using two commonly extemporaneously compounded formulations, the authors demonstrated the application of this tool. Advantages and limitations of the ultraviolet-visible technique are discussed. The authors speculate that more advanced spectral techniques for quality control will be adopted in the future. These techniques will be more accurate and will be associated with fewer limitations. However, costs associated with use will be greater. PMID:26625572

  19. Simple, rapid detection of influenza A (H1N1) viruses using a highly sensitive peptide-based molecular beacon.

    PubMed

    Lim, Eun-Kyung; Guk, Kyeonghye; Kim, Hyeran; Chung, Bong-Hyun; Jung, Juyeon

    2016-01-01

    A peptide-based molecular beacon (PEP-MB) was prepared for the simple, rapid, and specific detection of H1N1 viruses using a fluorescence resonance energy transfer (FRET) system. The PEP-MB exhibited minimal fluorescence in its "closed" hairpin structure. However, in the presence of H1N1 viruses, the specific recognition of the hemagglutinin (HA) protein of H1 strains by the PEP-MB causes the beacon to assume an "open" structure that emits strong fluorescence. The PEP-MB could detect H1N1 viruses within 15 min or even 5 min and can exhibit strong fluorescence even at low viral concentrations, with a detection limit of 4 copies. PMID:26509476

  20. Simple and rapid green synthesis of micrometer scale single crystalline gold nanoplates using chitosan as the reducing agent

    NASA Astrophysics Data System (ADS)

    Alex, Saji; Tian, Kun; Teng, Shiang; Siegel, Gene; Tiwari, Ashutosh

    2014-11-01

    A simple, rapid and green chemical method for the synthesis of single crystalline gold nanoplates of several micrometeres in size has been demonstrated. The synthesis involved the reduction of HAuCl4 in aqueous solution using low molecular weight chitosan at boiling temperature for 25 min. The [Au3+]:[chitosan] molar ratio plays an important role in the formation of gold nanoplates and found that an optimized molar ratio in the range of 80 to 125 was suitable for the formation of nanoplates. The size and morphology of the nanoplates can be tuned by adjusting the molar ratio. In this process, the chitosan functions both as a reducing as well as a stabilizing agent and no other special agents were added to induce the nanoplate formation. The obtained nanoplates were single crystals with (1 1 1) planes as the basal planes with shapes of hexagonal, triangular, or truncated triangular plates.

  1. Simple and rapid screening procedure for 143 new psychoactive substances by liquid chromatography-tandem mass spectrometry.

    PubMed

    Adamowicz, Piotr; Tokarczyk, Bogdan

    2016-07-01

    In recent years, many new psychoactive substances (NPS) from several drug classes have appeared on the drug market. These substances, also known as 'legal highs', belong to different chemical classes. Despite the increasing number of NPS, there are few comprehensive screening methods for their detection in biological specimens. In this context, the purpose of this study was to develop a fast and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening procedure for NPS in blood. The elaborated method allows the simultaneous screening of 143 compounds from different groups (number of compounds): cathinones (36), phenethylamines (26), tryptamines (18), piperazines (9), piperidines (2), synthetic cannabinoids (34), arylalkylamines (7), arylcyclohexylamines (3), aminoindanes (2), and other drugs (6). Blood samples (0.2 mL) were precipitated with acetonitrile (0.6 mL). The separation was achieved with gradient mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water in 14 min. Detection of all compounds was based on multiple reaction monitoring (MRM) transitions. The total number of transitions monitored in dynamic mode was 432. The whole procedure was rapid and simple. The limits of detection (LODs) estimated for 104 compounds were in the range 0.01-3.09 ng/mL. The extraction recoveries determined for 32 compounds were from 1.8 to 133%. The procedure was successfully applied to the analysis of forensic blood samples in routine casework. The developed method should have wide applicability for rapid screening of new drugs of abuse in forensic or clinical samples. The procedure can be easily expanded for more substances. Copyright © 2015 John Wiley & Sons, Ltd. PMID:25976069

  2. Simple and Rapid Determination of Ferulic Acid Levels in Food and Cosmetic Samples Using Paper-Based Platforms

    PubMed Central

    Tee-ngam, Prinjaporn; Nunant, Namthip; Rattanarat, Poomrat; Siangproh, Weena; Chailapakul, Orawon

    2013-01-01

    Ferulic acid is an important phenolic antioxidant found in or added to diet supplements, beverages, and cosmetic creams. Two designs of paper-based platforms for the fast, simple and inexpensive evaluation of ferulic acid contents in food and pharmaceutical cosmetics were evaluated. The first, a paper-based electrochemical device, was developed for ferulic acid detection in uncomplicated matrix samples and was created by the photolithographic method. The second, a paper-based colorimetric device was preceded by thin layer chromatography (TLC) for the separation and detection of ferulic acid in complex samples using a silica plate stationary phase and an 85:15:1 (v/v/v) chloroform: methanol: formic acid mobile phase. After separation, ferulic acid containing section of the TLC plate was attached onto the patterned paper containing the colorimetric reagent and eluted with ethanol. The resulting color change was photographed and quantitatively converted to intensity. Under the optimal conditions, the limit of detection of ferulic acid was found to be 1 ppm and 7 ppm (S/N = 3) for first and second designs, respectively, with good agreement with the standard HPLC-UV detection method. Therefore, these methods can be used for the simple, rapid, inexpensive and sensitive quantification of ferulic acid in a variety of samples. PMID:24077320

  3. Robust Polypropylene Fabrics Super-Repelling Various Liquids: A Simple, Rapid and Scalable Fabrication Method by Solvent Swelling.

    PubMed

    Zhu, Tang; Cai, Chao; Duan, Chunting; Zhai, Shuai; Liang, Songmiao; Jin, Yan; Zhao, Ning; Xu, Jian

    2015-07-01

    A simple, rapid (10 s) and scalable method to fabricate superhydrophobic polypropylene (PP) fabrics is developed by swelling the fabrics in cyclohexane/heptane mixture at 80 °C. The recrystallization of the swollen macromolecules on the fiber surface contributes to the formation of submicron protuberances, which increase the surface roughness dramatically and result in superhydrophobic behavior. The superhydrophobic PP fabrics possess excellent repellency to blood, urine, milk, coffee, and other common liquids, and show good durability and robustness, such as remarkable resistances to water penetration, abrasion, acidic/alkaline solution, and boiling water. The excellent comprehensive performance of the superhydrophobic PP fabrics indicates their potential applications as oil/water separation materials, protective garments, diaper pads, or other medical and health supplies. This simple, fast and low cost method operating at a relatively low temperature is superior to other reported techniques for fabricating superhydrophobic PP materials as far as large scale manufacturing is considered. Moreover, the proposed method is applicable for preparing superhydrophobic PP films and sheets as well. PMID:26061028

  4. A Simple and Rapid Procedure for the Detection of Genes Encoding Shiga Toxins and Other Specific DNA Sequences

    PubMed Central

    Nejman-Faleńczyk, Bożena; Bloch, Sylwia; Januszkiewicz, Aleksandra; Węgrzyn, Alicja; Węgrzyn, Grzegorz

    2015-01-01

    A novel procedure for the detection of specific DNA sequences has been developed. This procedure is based on the already known method employing PCR with appropriate primers and a sequence-specific DNA probe labeled with the fluorescent agent 6-carboxylfluorescein (FAM) at the 5′ end and the fluorescence quencher BHQ-1 (black hole quencher) at the 3′ end. However, instead of the detection of the fluorescence signal with the use of real-time PCR cyclers, fluorescence/luminescence spectrometers or fluorescence polarization readers, as in all previously-reported procedures, we propose visual observation of the fluorescence under UV light directly in the reaction tube. An example for the specific detection of the Shiga toxin-producing Escherichia coli (STEC) strains, by detecting Shiga toxin genes, is demonstrated. This method appears to be specific, simple, rapid and cost effective. It may be suitable for use in research laboratories, as well as in diagnostic units of medical institutions, even those equipped only with a thermocycler and a UV transilluminator, particularly if rapid identification of a pathogen is required. PMID:26580652

  5. A simple and rapid procedure for the detection of genes encoding Shiga toxins and other specific DNA sequences.

    PubMed

    Nejman-Faleńczyk, Bożena; Bloch, Sylwia; Januszkiewicz, Aleksandra; Węgrzyn, Alicja; Węgrzyn, Grzegorz

    2015-11-01

    A novel procedure for the detection of specific DNA sequences has been developed. This procedure is based on the already known method employing PCR with appropriate primers and a sequence-specific DNA probe labeled with the fluorescent agent 6-carboxylfluorescein (FAM) at the 5' end and the fluorescence quencher BHQ-1 (black hole quencher) at the 3' end. However, instead of the detection of the fluorescence signal with the use of real-time PCR cyclers, fluorescence/luminescence spectrometers or fluorescence polarization readers, as in all previously-reported procedures, we propose visual observation of the fluorescence under UV light directly in the reaction tube. An example for the specific detection of the Shiga toxin-producing Escherichia coli (STEC) strains, by detecting Shiga toxin genes, is demonstrated. This method appears to be specific, simple, rapid and cost effective. It may be suitable for use in research laboratories, as well as in diagnostic units of medical institutions, even those equipped only with a thermocycler and a UV transilluminator, particularly if rapid identification of a pathogen is required. PMID:26580652

  6. HIV/AIDS

    MedlinePlus

    ... HIV infections. HIV infection is often diagnosed through rapid diagnostic tests (RDTs), which detect the presence or absence of ... accuracy. It is important to note that serological tests detect antibodies produced ... pathogens, rather than direct detection of HIV itself. Most ...

  7. A rapid assessment of post-disclosure experiences of urban HIV-positive and HIV-negative school-aged children in Kenya

    PubMed Central

    2015-01-01

    There has been limited involvement of HIV-negative children in HIV disclosure studies; most studies conducted on the effects of disclosure on children have been with HIV-positive children and HIV-positive mother-child dyads. Seven HIV-positive and five HIV-negative children participated in a larger study conducted to understand the lived experiences of HIV-positive parents and their children during the disclosure process in Kenya. In this study, the experiences of these 12 children after receiving disclosure of their own and their parents’ illnesses respectively are presented. Each child underwent an in-depth qualitative semi-structured digitally recorded interview. The recorded interviews were transcribed and loaded into NVivo8 for phenomenological data analysis. Five themes emerged from the data, indicating that HIV-positive and negative children appear to have differing post-disclosure experiences revolving around acceptance of illness, stigma and discrimination, medication consumption, sexual awareness, and use of coping mechanisms. Following disclosure, HIV-negative children accepted their parents’ illnesses within a few hours to a few weeks; HIV-positive children took weeks to months to accept their own illnesses. HIV-negative children knew of high levels of stigma and discrimination within the community; HIV-positive children reported experiencing indirect incidences of stigma and discrimination. HIV-negative children wanted their parents to take their medications, stay healthy, and pay their school fees so they could have a better life in the future; HIV-positive children viewed medication consumption as an ordeal necessary to keep them healthy. HIV-negative children wanted their parents to speak to them about sexual-related matters; HIV-positive children had lingering questions about relationships, use of condoms, marriage, and childbearing options. All but one preadolescent HIV-positive child had self-identified a person to speak with for social

  8. A rapid assessment of post-disclosure experiences of urban HIV-positive and HIV-negative school-aged children in Kenya.

    PubMed

    Gachanja, Grace

    2015-01-01

    There has been limited involvement of HIV-negative children in HIV disclosure studies; most studies conducted on the effects of disclosure on children have been with HIV-positive children and HIV-positive mother-child dyads. Seven HIV-positive and five HIV-negative children participated in a larger study conducted to understand the lived experiences of HIV-positive parents and their children during the disclosure process in Kenya. In this study, the experiences of these 12 children after receiving disclosure of their own and their parents' illnesses respectively are presented. Each child underwent an in-depth qualitative semi-structured digitally recorded interview. The recorded interviews were transcribed and loaded into NVivo8 for phenomenological data analysis. Five themes emerged from the data, indicating that HIV-positive and negative children appear to have differing post-disclosure experiences revolving around acceptance of illness, stigma and discrimination, medication consumption, sexual awareness, and use of coping mechanisms. Following disclosure, HIV-negative children accepted their parents' illnesses within a few hours to a few weeks; HIV-positive children took weeks to months to accept their own illnesses. HIV-negative children knew of high levels of stigma and discrimination within the community; HIV-positive children reported experiencing indirect incidences of stigma and discrimination. HIV-negative children wanted their parents to take their medications, stay healthy, and pay their school fees so they could have a better life in the future; HIV-positive children viewed medication consumption as an ordeal necessary to keep them healthy. HIV-negative children wanted their parents to speak to them about sexual-related matters; HIV-positive children had lingering questions about relationships, use of condoms, marriage, and childbearing options. All but one preadolescent HIV-positive child had self-identified a person to speak with for social support

  9. Interferon-induced transmembrane protein-3 rs12252-C is associated with rapid progression of acute HIV-1 infection in Chinese MSM cohort

    PubMed Central

    Zhang, Yonghong; Makvandi-Nejad, Shokouh; Qin, Ling; Zhao, Yan; Zhang, Tong; Wang, Lili; Repapi, Emmanouela; Taylor, Stephen; McMichael, Andrew; Li, Ning; Dong, Tao; Wu, Hao

    2015-01-01

    Background: The interferon-inducible transmembrane protein-3 (IFITM3) is a protein that restricts multiple pathogenic viruses such as influenza virus. The single-nucleotide polymorphism rs12252-C, which is rare in Caucasian populations, but much more common in the Han Chinese population, has been found in much higher homozygous frequency in patients with severe acute influenza. Until now, there has been no study on the effect of this genetic variant on the clinical control of other viral infections. Objectives: To investigate the impact of IFITM3-rs12252 genotypes on primary HIV-1 infection progression in an acute HIV-1-infected cohort in Beijing (PRIMO), China. Design and methods: We identified IFITM3-rs12252 genotypes of 178 acute HIV-1-infected patients and 196 HIV-negative candidates from the PRIMO cohort. HIV-1 viral load and CD4+ T-cell counts were monitored at multiple time points during the first year of infection, and the association between IFITM3-rs12252 genotype and disease progression was evaluated. Results: The current study shows that the IFITM3-rs12252 genetic variant affects the progression of HIV-1 infection, but not the acquisition. A significantly higher frequency of the CC/CT genotypes was found in rapid progressors compared to nonprogressors. Patients with CC/CT genotypes showed an elevated peak viremia level and significantly lower CD4+ T-cell count at multiple time points during the first year of primary infection, and a significantly higher risk of rapid decline of the CD4+ T-cell count to below 350 cells/μl. Conclusion: A novel association between IFITM3 gene polymorphism and rapid disease progression is reported in an acute HIV-1-infected MSM cohort in China. PMID:25784441

  10. A simple and rapid chromatographic method to determine unauthorized basic colorants (rhodamine B, auramine O, and pararosaniline) in processed foods.

    PubMed

    Tatebe, Chiye; Zhong, Xining; Ohtsuki, Takashi; Kubota, Hiroki; Sato, Kyoko; Akiyama, Hiroshi

    2014-09-01

    A simple and rapid high-performance liquid chromatography (HPLC) method to determine basic colorants such as pararosaniline (PA), auramine O (AO), and rhodamine B (RB) in various processed foods was developed. Linearity of the calibration curves ranged from 0.05 to 50 μg/mL for PA and 0.05-100 μg/mL for AO and RB. The detection and quantification limits (LOD and LOQ) of the basic colorants, which were evaluated as signal-to-noise ratios of 3 for LOD and 10 for LOQ, ranged from 0.0125 to 0.05 and 0.025 to 0.125 μg/g, respectively. The recoveries and relative standard deviations of three basic colorants in six processed foods, namely, chili sauce, curry paste, gochujang (hot pepper paste), tandoori chicken (roasted chicken prepared with yogurt and spices), powder soup, and shrimp powder ranged from 70.2% to 102.8% and 0.8% to 8.0%, respectively. The intraday precision of the recovery test ranged from 1.7% to 4.5%, whereas the interday precision ranged from 3.7% to 7.7%. The reported method has been successfully applied to basic colorant determination in various processed foods such as fat-based food matrices (curry paste and tandoori chicken), chili products (gochujang and chili sauce), and protein-based products (shrimp powder and powder soup). Thin layer chromatography and liquid chromatography/mass spectrometry methods for the determination of basic colorants in processed foods were also developed for rapid analysis and identification, respectively. These methods are very useful for monitoring unauthorized basic colorants in inspection centers or quarantine laboratories in many countries. PMID:25473512

  11. A simple and rapid chromatographic method to determine unauthorized basic colorants (rhodamine B, auramine O, and pararosaniline) in processed foods

    PubMed Central

    Tatebe, Chiye; Zhong, Xining; Ohtsuki, Takashi; Kubota, Hiroki; Sato, Kyoko; Akiyama, Hiroshi

    2014-01-01

    A simple and rapid high-performance liquid chromatography (HPLC) method to determine basic colorants such as pararosaniline (PA), auramine O (AO), and rhodamine B (RB) in various processed foods was developed. Linearity of the calibration curves ranged from 0.05 to 50 μg/mL for PA and 0.05–100 μg/mL for AO and RB. The detection and quantification limits (LOD and LOQ) of the basic colorants, which were evaluated as signal-to-noise ratios of 3 for LOD and 10 for LOQ, ranged from 0.0125 to 0.05 and 0.025 to 0.125 μg/g, respectively. The recoveries and relative standard deviations of three basic colorants in six processed foods, namely, chili sauce, curry paste, gochujang (hot pepper paste), tandoori chicken (roasted chicken prepared with yogurt and spices), powder soup, and shrimp powder ranged from 70.2% to 102.8% and 0.8% to 8.0%, respectively. The intraday precision of the recovery test ranged from 1.7% to 4.5%, whereas the interday precision ranged from 3.7% to 7.7%. The reported method has been successfully applied to basic colorant determination in various processed foods such as fat-based food matrices (curry paste and tandoori chicken), chili products (gochujang and chili sauce), and protein-based products (shrimp powder and powder soup). Thin layer chromatography and liquid chromatography/mass spectrometry methods for the determination of basic colorants in processed foods were also developed for rapid analysis and identification, respectively. These methods are very useful for monitoring unauthorized basic colorants in inspection centers or quarantine laboratories in many countries. PMID:25473512

  12. Rapid (1 hour) high performance gel filtration chromatography resolves coexisting simple micelles, mixed micelles, and vesicles in bile.

    PubMed

    Cohen, D E; Carey, M C

    1990-11-01

    We describe the use and validation of Superose 6, a high performance gel filtration medium for rapid, high resolution separation and sizing of coexisting simple micelles, mixed micelles, and vesicles in bile. We fractionated model biles (1.7-4.2 g/dl total lipid concentration, 0.15 M NaCl) composed of lecithin (L), cholesterol (Ch), and the common bile salt taurocholate (TC) using Superose 6 gel filtration columns (1.0 cm diameter, 30 cm length, 0.5 ml model bile application, 1.0 ml fractions) pre-equilibrated and eluted with 2.5-10.0 mM TC. Lipid particle sizes were determined by quasielastic light scattering and lipid compositions by conventional analyses. In the absence of L and Ch, pure TC "biles" (32.2 mM), when eluted in the presence of 7.5 mM TC, yielded a single peak of particles (mean hydrodynamic radii, Rh values of 11-15 A), consistent with simple TC micelles. Model biles containing L and TC ([L] = 13.8 mM, [TC] = 32.2 mM) were fractionated with baseline resolution into TC-L mixed micelles, (Rh values of 30-40 A) and simple TC micelles. In agreement with the ternary TC-L-H2O phase diagram (Mazer, N. A., et al. 1980. Biochemistry. 19: 601-615), the proportions of simple and mixed micelles were inversely related to L concentrations ([L] = 0-32.2 mM) and correlated positively with eluant TC concentration. Superose 6 gel fractionation of model biles "super-saturated" with Ch (TC:L:Ch molar ratio 27:63:10, total lipid concentration 3 g/dl) yielded high resolution separation of vesicles (Rh value of 320 A) from mixed micelles of TC-L-Ch (Rh values of 40-50 A) and simple TC micelles (Rh values of 11-15 A). At an eluant TC concentration of 7.5 mM, Ch-rich vesicles (Ch/L molar ratio = 1.6) separated that contained 40% of total Ch, 9% of total L, and no TC, accurately reflecting predictions of the quaternary L-Ch-TC-H2O metastable phase diagram (Mazer, N. A., and M. C. Carey. 1983 Biochemistry. 22: 426-442). This suggested that a 7.5 mM TC concentration

  13. H sup + -ATP synthase from rat liver mitochondria. A simple, rapid purification method of the functional complex and its characterization

    SciTech Connect

    Yoshihara, Yutaka; Nagase, Hideki; Yamane, Takeshi; Oka, Hideki; Tani, Isamu; Higuti, Tomihiko )

    1991-07-16

    A novel, simple, and rapid preparative method for purification of rat liver H{sup +}-ATP synthase by anion-exchange HPLC was developed. The H{sup +}-ATP synthase purified had higher ATPase activity in the absence of added phospholipids than any preparation reported previously, and this activity was completely inhibited by oligomycin. When reconstituted into proteoliposomes, the H{sup +}-ATP synthase showed an ATP-dependent 8-anilinonaphthalene-1-sulfonate response and ATP-P{sub i} exchange activity, both of which were also completely inhibited by oligomycin and an uncoupler, indicating the intactness of the H{sup +}-ATP synthase. An immunochemical study and a labeling experiment with N,N{prime}-({sup 14}C)dicyclohexylcarbondiimide (({sup 14}C)DCCD) demonstrated the presence of chargerin II (a product of mitochondrial A6L DNA) and DCCD-binding protein (subunit c) in the complex. The subunits of the complex were separated into 11 main fractions by reverse-phase HPLC, and 3 of them and the {sigma} subunit in F{sub 1} were partially sequenced. A search for sequence homologies indicated that these components were subunit b, coupling factor 6, subunit {sigma}, and subunit e. This is the first report of the existence of subunit b, factor 6, and chargerin II in K{sup +}-ATP synthase purified from rat liver mitochondria.

  14. The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue.

    PubMed

    Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

    2016-01-01

    In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15-150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method. PMID:27282822

  15. The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

    PubMed Central

    Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

    2016-01-01

    In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15–150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method. PMID:27282822

  16. Simple and rapid solid-phase radioimmunoassay for serum progesterone, using the protein A of Staphylococcus aureus as immunoadsorbent

    SciTech Connect

    Jungers, J.; Delogne-Desnoeck, J.; Robyn, C.

    1981-07-01

    A simple, rapid, and inexpensive radioimmunoassay method for serum progesterone is described, which uses a solid-phase technique for separation of antibody-bound from antibody-free progesterone. Rabbit antiprogesterone immunoglobulins are adsorbed on the protein A of formaldehyde- and heat-treated Staphylococcus aureus cells (Pansorbin; Calbiochem-Behring Corp., La Jolla, California). The suspension of antibody-coated Pansorbin retains all its binding activity of 1-2-H(N)-progesterone when kept at + 4/sup 0/ or at -25/sup 0/C for at least 4 months. Dose-response curves obtained with ether-serum extracts and with the progesterone standard do not deviate significantly from parallelism. The progesterone standard gives identical dose-response curves whether diluted in the assay buffer or in a progesterone-free ether-serum extract. The sensitivity of the assay is 0.02 ng/assay tube. The intra-assay variation coefficient is 16%, and the routine interassay variation coefficient is 17%. The mean serum progesterone concentrations were 0.55 ng/ml during the follicular phase of the menstrual cycle and 12.5 ng/ml during the luteal phase. The average blank value for distilled water was 0.02 ng/assay tube.

  17. New simple and rapid method for purification of mesenchymal stem cells from the human umbilical cord Wharton jelly.

    PubMed

    Montanucci, Pia; Basta, Giuseppe; Pescara, Teresa; Pennoni, Ilaria; Di Giovanni, Francesca; Calafiore, Riccardo

    2011-11-01

    We have developed a simple and rapid method for isolation of human umbilical cord matrix stem cells (hUCMS). The umbilical cord contains a virtual inexhaustible source of adult stem cells. We have substantially modified, simplified, and improved previously reported hUCMS isolation procedures in terms of either used enzyme type, or digestion time, and substantially enhanced the final product yield and purity. The isolated hUCMS were positive for CD90, CD117, and SCF, and negative for CD31 and CD45 surface markers. mRNA and related proteins (i.e., Sox2, Oct4a, Nanog, ABCG2, and c-Myc) that coincide with an uncommitted cell status also were detected. hUCMS express genes and proteins for CD90 and Nestin that are associated with mesenchymal stem cells, as well as other genes that specifically relate to different embryonic germ layers, namely, Vimentin, Sox7, Sox17, FoxA2, E-cadherin, and N-cadherin. hUCMS showed multilineage cell differentiation potential into adipogenic, osteogenic, and neural cell phenotypes, under the influence of lineage-specific, differentiation culture media. Moreover, the basal expression of endocrine cell markers makes these cells seemingly suitable for endocrine cell phenotype differentiation. Noteworthy, Activin A induced hUCMS to acquire definitive endoderm cell markers. PMID:21679124

  18. Rapid and simple method to determine morphine and its metabolites in rat plasma by liquid chromatography-mass spectrometry.

    PubMed

    Projean, Denis; Minh Tu, The; Ducharme, Julie

    2003-04-25

    A rapid and simple method for the determination of morphine (M), normorphine (NM), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in plasma by high-performance liquid chromatographic separation with mass spectrometric detection (HPLC-MS) has been developed. Samples (40 microl) were cleaned-up by protein precipitation with two volumes (80 microl) of acetonitrile and reconstituted in formic acid 0.1% in water. Naloxone was used as internal standard. Analytes were separated on a phenyl-hexyl column using a step-gradient (1 ml/min) of acetonitrile and formic acid in water. Acetonitrile was added post-column (0.3 ml/min). Quantification of morphine and its metabolites was achieved with an Agilent 1100 series HPLC-MS system equipped with electrospray interface set to selected ion-monitoring (SIM) mode. Calibration curves covered a wide range of concentrations (2.44-10,000 nM) and were best fitted with a weighed quadratic equation. The limits of quantification achieved with this method were 2.44 nM for M and 4.88 nM for NM, M3G and M6G. The method proved accurate (85-98%), precise (C.V.<10%) and was successfully applied to a wide range of in vitro and in vivo pharmacokinetic studies in rodents. PMID:12650748

  19. The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

    NASA Astrophysics Data System (ADS)

    Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

    2016-06-01

    In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15–150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method.

  20. Analysis of a rapid, simple, and inexpensive technique used to obtain platelet-rich plasma for use in clinical practice.

    PubMed

    Rutkowski, James L; Thomas, Joseph M; Bering, C Larry; Speicher, Julie L; Radio, Nicholas M; Smith, Douglas M; Johnson, David A

    2008-01-01

    The use of platelet-rich plasma (PRP) has become more generally accepted, and implant dentists are using PRP more frequently to promote the healing of oral surgical and/or periodontal wounds. Critical elements of PRP are thought to be growth factors contained within the concentrated platelets. These growth factors are known to promote soft-tissue healing, angiogenesis and osteogenesis. We present a rapid, simple, and inexpensive methodology for preparing PRP using the Cliniseal centrifuge method. This study demonstrates that platelets are concentrated approximately 6-fold without altering platelet morphology. Further we demonstrate that key growth factors, platelet-derived growth factor BB (PDGF-BB), transforming growth factor B (TGF-B1), vasculature endothelial growth factor (VEGF), and epidermal growth factor (EGF) are present in comparable or higher concentrations than those reported with the use of other techniques. Prolonged bench set time (>3 hours) after centrifugation resulted in decreased concentration of TGF-B1 but not decreased concentration of PDGF-BB, VEGF, or EGF. This study confirms the molecular aspects of PRP obtained using this inexpensive and efficient methodology. PMID:18390240

  1. A simple and rapid method for monitoring dissolved oxygen in water with a submersible microbial fuel cell (SBMFC).

    PubMed

    Zhang, Yifeng; Angelidaki, Irini

    2012-01-01

    A submersible microbial fuel cell (SBMFC) was developed as a biosensor for in situ and real time monitoring of dissolved oxygen (DO) in environmental waters. Domestic wastewater was utilized as a sole fuel for powering the sensor. The sensor performance was firstly examined with tap water at varying DO levels. With an external resistance of 1000Ω, the current density produced by the sensor (5.6 ± 0.5-462.2 ± 0.5 mA/m(2)) increased linearly with DO level up to 8.8 ± 0.3mg/L (regression coefficient, R(2)=0.9912), while the maximum response time for each measurement was less than 4 min. The current density showed different response to DO levels when different external resistances were applied, but a linear relationship was always observed. Investigation of the sensor performance at different substrate concentrations indicates that the organic matter contained in the domestic wastewater was sufficient to power the sensing activities. The sensor ability was further explored under different environmental conditions (e.g. pH, temperature, conductivity, and alternative electron acceptor), and the results indicated that a calibration would be required before field application. Lastly, the sensor was tested with different environmental waters and the results showed no significant difference (p>0.05) with that measured by DO meter. The simple, compact SBMFC sensor showed promising potential for direct, inexpensive and rapid DO monitoring in various environmental waters. PMID:22726635

  2. Simple and rapid high-performance liquid chromatographic method for the determination of aspartame and its metabolites in foods.

    PubMed

    Gibbs, B F; Alli, I; Mulligan, C N

    1996-02-23

    A method for the determination of aspartame (N-L-alpha-aspartyl-L-phenylalanine methyl ester) and its metabolites, applicable on a routine quality assurance basis, is described. Liquid samples (diet Coke, 7-Up, Pepsi, etc.) were injected directly onto a mini-cartridge reversed-phase column on a high-performance liquid chromatographic system, whereas solid samples (Equal, hot chocolate powder, pudding, etc.) were extracted with water. Optimising chromatographic conditions resulted in resolved components of interest within 12 min. The by-products were confirmed by mass spectrometry. Although the method was developed on a two-pump HPLC system fitted with a diode-array detector, it is straightforward and can be transformed to the simplest HPLC configuration. Using a single-piston pump (with damper), a fixed-wavelength detector and a recorder/integrator, the degradation of products can be monitored as they decompose. The results obtained were in harmony with previously reported tedious methods. The method is simple, rapid, quantitative and does not involve complex, hazardous or toxic chemistry. PMID:8900578

  3. A simple and rapid strategy for the molecular cloning and monitoring of mouse HtrA2 serine protease.

    PubMed

    Kim, Goo-Young; Nam, Min-Kyung; Kim, Sang-Soo; Kim, Ho-Young; Lee, Sang-Kyu; Rhim, Hyangshuk

    2008-03-01

    A simple and rapid strategy for molecular cloning using a gel-free and antibiotic selection method is described which allows for the complete elimination of DNA extraction by gel electrophoresis, and thus has several advantages over gel-based cloning methods, including: (i) a cloning efficiency that is approximately 10-times higher due to the prevention of ethidium bromide ultraviolet-induced DNA damage and contamination with ligase inhibitors; (ii) the amount of plasmid DNA required is approximately five times less; and (iii) the cloning time is several hours less. Once the target gene, such as mouse HtrA2 serine protease, was cloned into the pEGFP-N3 plasmid, the integrity of the kanamycin-resistant molecular clone encoding the GFP fusion protein was verified by immunoblot and immunofluorescence assays. In addition, the integrity of the ampicillin-resistant molecular clone was directly evaluated by analyzing the expression and affinity purification of the GST fusion protein and by measuring its enzymatic activity. Therefore, this method is suitable for the routine construction of a plasmid expressing the gene of interest, and the usefulness of this strategy can be demonstrated by monitoring the expression of the target gene in E. coli and mammalian cells. PMID:17939055

  4. White side test: A simple and rapid test for evaluation of nonspecific bacterial genital infections of repeat breeding cattle

    PubMed Central

    Bhat, Fayaz Ahmad; Bhattacharyya, Hiranya Kumar; Hussain, Syed Akram

    2014-01-01

    The objective of the present study was to determine the grades of nonspecific bacterial infection of genitalia of repeat breeding cattle by a simple and rapid test under field condition. For this purpose, a total of 100 crossbred Jersey cows comprising of 80 repeat breeding animals presented for treatment and 20 normal cyclic (control group) animals presented for artificial insemination at their first service were selected. Estrual cervical mucus from all the animals was collected at 8 to 12 hr after the onset of behavioral estrus and subjected to white side test (WST) and bacteriological examination. The results of WST showed only 15% of control group had infection but the remaining 85% were free of it. In contrast, the majority of repeat breeding animals (57/80) showed infection (71.25%) and only 28.75% animals were free of infection. In bacterial culture, 60 (75.00%) from the 80 repeat breeding animals were found positive, and 20 (25.00%) were free of bacteria. All the three samples of control group that showed no color reaction in WST had also no growth in bacterial culture. The WST results showed a positive (p < 0.01) correlation of 0.48 with bacterial culture. It is thus concluded that under field condition WST can be used as a prime modality for ascertaining nonspecific bacterial infection of repeat breeding cattle before subjecting them to any antibiotic therapy thereby reducing the cost of diagnosis and treatment. PMID:25568715

  5. Sex isn't that simple: culture and context in HIV prevention interventions for gay and bisexual male adolescents.

    PubMed

    Harper, Gary W

    2007-11-01

    Gay and bisexual male adolescents and young adults in the United States have been disproportionately impacted by the HIV pandemic. Despite the steadily increasing rise in their HIV infection rates, there has not been a commensurate increase in HIV prevention programs targeted to the unique social and sexual lives of these youths. Programs that address cultural and contextual factors that influence sexual risk and protective behaviors need to be developed, implemented, and rigorously evaluated. These interventions should address the potential influences of sexual and gay culture on the HIV risk/protective behaviors of gay and bisexual adolescents, as well as the influence of more traditional cultural factors related to ethnicity. The influence of contextual developmental factors should also be addressed. This may include an incorporation into prevention programs of the societal-level influences of heterosexism and masculinity ideology and the individual-level influences of sexual identity and ethnic identity development. Researchers and interventionists need to be creative and innovative in their HIV prevention approaches and ensure that programs are grounded in the lives and realities of gay and bisexual adolescents and young adults. PMID:18020756

  6. Applying qualitative data derived from a Rapid Assessment and Response (RAR) approach to develop a community-based HIV prevention program for adolescents in Thailand.

    PubMed

    Watthayu, Nantiya; Wenzel, Jennifer; Panchareounworakul, Kobkul

    2015-01-01

    HIV education programs are needed to address risk-taking behavior for adolescents. The purpose of our study was to use the World Health Organization's Rapid Assessment and Response (RAR) method to design a community-based, cultural- and age-appropriate HIV prevention program for adolescents in Bangkok, Thailand. Adolescent single-gender-specific focus groups (n = 3; 28 participants) were used to gather reactions/ideas about program topics/approaches. An adult, mixed-gender group was held to review information identified by adolescents. Sessions were audiotaped and transcribed verbatim. Themes regarding HIV content and the process of implementation emerged from a qualitative content analysis of the data. Community representatives recommended incorporation of HIV information and risk-prevention skills. Information delivery suggestions included small group discussions, interactive games/role-playing, program materials/terminology, and HIV-infected program facilitators. Community members provided critical input toward an HIV prevention program tailored to meet adolescents' unique needs/interests. The RAR model provides opportunities to engage communities in developing health-related interventions. PMID:26279387

  7. Case finding advantage of HIV rapid tests in community settings: men who have sex with men in 12 programme areas in China, 2011.

    PubMed

    Zhang, Dapeng; Qi, Jinlei; Fu, Xiaojing; Meng, Sining; Li, Chengmei; Sun, Jiangping

    2015-05-01

    We sought to describe the advantage of rapid tests over ELISA tests in community-based screening for HIV among men who have sex with men (MSM) in urban areas of China. Data of 31,406 screening tests conducted over six months in 2011 among MSM across 12 areas were analyzed to compare the differences between those receiving rapid testing and ELISA. Rapid tests accounted for 45.8% of these screening tests. The rate of being screened positive was 7.2% among rapid tests and 5.3% for ELISA tests (χ(2 )= 49.161, p < 0.001). This advantage of rapid test in HIV case finding persisted even when socio-demographic, behavioural, screening recruitment channel and city were controlled for in logistic regression (exp[beta] = 1.42, p < 0.001, 95% CI = 1.27,1.59). MSM who received rapid tests, compared with those tested by ELISA, were less likely to use condoms during last anal sex (50.8% vs. 72.3%, χ(2 )= 1706.146, p < 0.001), more likely to have multiple sex partners (55.7% vs. 49.5%, χ(2 )= 238.188, p < 0.001) and less likely to have previously undergone HIV testing (38.8% vs. 54.7%, χ(2 )= 798.476, p < 0.001). These results demonstrate the robustness of the advantage of rapid tests over traditional ELISA tests in screening for MSM with HIV infection in cooperation with community-based organizations in urban settings in China. PMID:25028452

  8. Rapid HIV antibody testing among men who have sex with men who visited a gay bathhouse in Hangzhou, China: a cross-sectional study

    PubMed Central

    Ma, Qiaoqin; Xia, Shichang; Pan, Xiaohong; Cai, Gaofeng; Zhou, Xin; Wang, Hui; Peng, Zhihang

    2015-01-01

    Objective To understand the prevalence and correlates of rapid HIV antibody testing (RHT) among men who have sex with men (MSM) clients of gay bathhouses. Design Cross-sectional questionnaire survey. Setting This study was conducted in a gay bathhouse in Hangzhou, China. Participants 354 MSM were validly recruited from October to December 2012. Inclusion criteria were (1) men who visited the gay bathhouse, (2) men who had engaged in sex with men during the previous 6 months, (3) first-time participants in this survey and (4) men who were HIV-negative if already tested. Measures Sociodemographic measures included factors related to sexual behaviour and HIV risk perception, and the scales of HIV-related knowledge and behavioural intervention that each participant received. Results Of the 354 participants, 222 (62.7%) were rapid tested during the previous 6 months; of them, 66.2% were tested at the Centers for Disease Prevention and Control (CDC), and 46.8% at gay venues. The following factors were independently associated with rapid testing within the previous 6 months: sexual initiation at 20–29 years of age, ever having undergone standard testing, ever having seen a sexually transmitted disease doctor, consistent use of condom during the past 6 months, familiarity with RHT and perception of possible HIV infection. Conclusions Publicity of RHT and risk education for HIV infection are necessary to promote RHT among MSM who visit gay bathhouses. The characteristics of sexual behaviours among those who do and do not undergo RHT should be taken into consideration while promoting the service in this group. PMID:26346876

  9. A simple and rapid method for calixarene-based selective extraction of bioactive molecules from natural products.

    PubMed

    Segneanu, Adina-Elena; Damian, Daniel; Hulka, Iosif; Grozescu, Ioan; Salifoglou, Athanasios

    2016-03-01

    Natural products derived from medicinal plants have gained an important role in drug discovery due to their complex and abundant composition of secondary metabolites, with their structurally unique molecular components bearing a significant number of stereo-centers exhibiting high specificity linked to biological activity. Usually, the extraction process of natural products involves various techniques targeting separation of a specific class of compounds from a highly complex matrix. Aiding the process entails the use of well-defined and selective molecular extractants with distinctly configured structural attributes. Calixarenes conceivably belong to that class of molecules. They have been studied intensely over the years in an effort to develop new and highly selective receptors for biomolecules. These macrocycles, which display remarkable structural architectures and properties, could help usher a new approach in the efficient separation of specific classes of compounds from complex matrices in natural products. A simple and rapid such extraction method is presented herein, based on host-guest interaction(s) between a calixarene synthetic receptor, 4-tert-butyl-calix[6]arene, and natural biomolecular targets (amino acids and peptides) from Helleborus purpurascens and Viscum album. Advanced physicochemical methods (including GC-MS and chip-based nanoESI-MS analysis) suggest that the molecular structure and specifically the calixarene cavity size are closely linked to the nature of compounds separated. Incorporation of biomolecules and modification of the macrocyclic architecture during separation were probed and confirmed by scanning electronic microscopy and atomic force microscopy. The collective results project calixarene as a promising molecular extractant candidate, facilitating the selective separation of amino acids and peptides from natural products. PMID:26597796

  10. Rapid, simple and highly sensitive LC-ESI-MS/MS method for the quantification of tamsulosin in human plasma.

    PubMed

    Ramakrishna, N V S; Vishwottam, K N; Manoj, S; Koteshwara, M; Wishu, S; Varma, D P

    2005-12-01

    A simple, rapid, sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of tamsulosin (I), a highly selective alpha1-adrenoceptor antagonist used for the treatment of patients with symptomatic benign prostatic hyperplasia. The analyte and internal standard, mosapride (II) were extracted by liquid-liquid extraction with diethyl ether-dichloromethane (70:30, v/v) using a Glas-Col Multi-Pulse Vortexer. The chromatographic separation was performed on a reverse phase Waters symmetry C18 column with a mobile phase of 0.03% formic acid-acetonitrile (30:70, v/v). The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The mass transitions m/z 409.1 solidus in circle 228.1 and m/z 422.3 solidus in circle 198.3 were used to measure I and II, respectively. The assay exhibited a linear dynamic range of 0.1-50.0 ng/mL for tamsulosin in human plasma. The lower limit of quantitation was 100 pg/mL with a relative standard deviation of less than 10%. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 2.0 min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. PMID:15828055

  11. Simple and rapid RP-HPLC method for simultaneous determination of acyclovir and zidovudine in human plasma.

    PubMed

    Sharma, Megha; Nautiyal, Pragya; Jain, Surendra; Jain, Deepti

    2010-01-01

    Combination therapy with acyclovir and zidovudine is used for the treatment of herpes-infected immunocompromised patients. In the view of the optimal drug concentrations (minimum effective concentrations) for viral suppression and avoidance of drug toxicity, monitoring of drug levels has been considered essential to determine drug concentrations in plasma after administration of a dose of acyclovir and zidovudine. A simple, precise, and rapid RP-HPLC method has been developed for this purpose. Chromatographic separation was performed using methanol-water (50 + 50, v/v), pH 2.5 adjusted with orthophosphoric acid, as an isocratic mobile phase at a flow rate of 0.8 mL/min with an Inertsil ODS (C18) column (5 microm particle size, 250 x 4.60 mm id). Detection was carried out using a UV photo diode array detector at 258 nm. The plasma samples were prepared by a protein precipitation method. The retention time for acyclovir and zidovudine was 3.5 +/- 0.2 and 6.2 +/- 0.3 min, respectively. The method was linear in the range of 200-1800 and 400-3600 ng/mL with LOQ of 200 ng (SD = +/-1.4) and 400 ng (SD = +/-0.9) for zidovudine and acyclovir, respectively, in plasma. The mean accuracy was 98.0 and 96.4%, with average extraction recovery of 64.8 +/- 2.1 and 77.5 +/- 1.7% for lower nominal concentrations of acyclovir and zidovudine, respectively. PMID:21140658

  12. Simple, rapid zebrafish larva bioassay for assessing the potential of chemical pollutants and drugs to disrupt thyroid gland function.

    PubMed

    Raldúa, Demetrio; Babin, Patrick J

    2009-09-01

    Thyroid function may be altered by a very large number of chemicals routinely found in the environment Research evaluating potential thyroid disruption is ongoing, but there are thousands of synthetic and naturally occurring drugs and chemicals to be considered. European and United States policies call for the development of simple methodologies for screening endocrine-disrupting chemicals. Zebrafish are widely used as a model organism for assessing drug effects because of their small size, high fecundity, rapid organogenesis, morphological and physiological similarities to mammals, and easewithwhich large-scale phenotypic screening is performed. A zebrafish-based short-duration screening method was developed to detect the potential effect of chemicals and drugs on thyroid function. This method used a T4 immunofluorescence quantitative disruption test (TIQDT) to measure thyroid function. The 3 day exposure window protocol, from day 2 to day 5 postfertilization (dpf), avoided any potential side effects on thyroid gland morphogenesis. Methimazole, propylthiouracil, and potassium perchlorate, three well-known goitrogens, totally abolished T4 immunoreactivity in thyroid follicles in a dose-specific manner. Amiodarone, a human pharmaceutical with a reported cytotoxic effect on thyroid follicular cells, also decreased T4 levels. Moreover, exposure to 50 nM 3,3',5-triiodothyronine induced a significant decrease in T4 immunoreactivity as did DDT, 2,4-D, and 4-nonylphenol. In conclusion, these data indicated that TIQDT may be useful for obtaining initial information about the ability of environmental pollutants and drugs to impair thyroid gland function as well as assessing the combined effects of endocrine disruptors. PMID:19764258

  13. Rapid screening for Schistosoma mansoni in western Côte d'Ivoire using a simple school questionnaire.

    PubMed Central

    Utzinger, J.; N'Goran, E. K.; Ossey, Y. A.; Booth, M.; Traoré, M.; Lohourignon, K. L.; Allangba, A.; Ahiba, L. A.; Tanner, M.; Lengeler, C.

    2000-01-01

    The distribution of schistosomiasis is focal, so if the resources available for control are to be used most effectively, they need to be directed towards the individuals and/or communities at highest risk of morbidity from schistosomiasis. Rapid and inexpensive ways of doing this are needed, such as simple school questionnaires. The present study used such questionnaires in an area of western Côte d'Ivoire where Schistosoma mansoni is endemic; correctly completed questionnaires were returned from 121 out of 134 schools (90.3%), with 12,227 children interviewed individually. The presence of S. mansoni was verified by microscopic examination in 60 randomly selected schools, where 5047 schoolchildren provided two consecutive stool samples for Kato-Katz thick smears. For all samples it was found that 54.4% of individuals were infected with S. mansoni. Moreover, individuals infected with S. mansoni reported "bloody diarrhoea", "blood in stools" and "schistosomiasis" significantly more often than uninfected children. At the school level, Spearman rank correlation analysis showed that the prevalence of S. mansoni significantly correlated with the prevalence of reported bloody diarrhoea (P = 0.002), reported blood in stools (P = 0.014) and reported schistosomiasis (P = 0.011). Reported bloody diarrhoea and reported blood in stools had the best diagnostic performance (sensitivity: 88.2%, specificity: 57.7%, positive predictive value: 73.2%, negative predictive value: 78.9%). The study, which is probably the largest of its kind ever undertaken in Africa, revealed a moderate diagnostic performance of questionnaires for identifying individuals and/or communities at high risk from S. mansoni. PMID:10812739

  14. Identification of G-Quadruplex Inducers Usinga Simple, Inexpensiveand Rapid High Throughput Assay, and TheirInhibition of Human Telomerase

    PubMed Central

    Sassano, Maria Florencia; Schlesinger, Alexander P; Jarstfer, Michael B

    2012-01-01

    Telomeres are protein and DNA complexes located atchromosome ends. Telomeric DNA is composed of a double stranded region of repetitive DNA followed by single-stranded 3' extension of aG-rich sequence. Single-stranded G-rich sequencescan fold into G-quadruplex structures,and molecules that stabilize G-quadruplexes are known to inhibit the enzyme telomerase and disrupt telomere maintenance. Because telomere maintenance is required for proliferation of cancer cells, G-quadruplex stabilizers have become attractive prospects for anticancer drug discovery.However, telomere-targeting G-quadruplex ligands have yet to enter the clinic owing in part to poor pharmacokinetics and target selectivity. Increasing the pharmacophore diversity of G-quadruplex and specifically telomeric-DNA targeting agents should assist in overcoming these shortcomings. In this work, we report the identification and validation ofligands that bind telomeric DNA and induce G-quadruplex formationusing the NCI Diversity Set I, providing validation of anextremely simple, rapid and high-throughput screen using FRET technology. Hits from the screen were validated by examining telomerase inhibition and G-quadruplex inductionusing CD spectroscopy and DNA polymerase stop assays. We show that two known DNA binding molecules, ellipticine derivativeNSC 176327 (apyridocarbazole) and NSC 305831 (an antiparasitic hetero-cyclediamidine referred to as furamidine and DB75),are selective induceG-quadruplex formation in the human telomeric sequence and bind telomeric DNA quadruplexes in the absence of stabilizing monovalent cations with molar ratios(molecule: DNA)of 4:1and 1.5:1, respectively. PMID:23173022

  15. [Evaluation of Basic Performance of "Point Strip ferritin-3000" for Simple and Rapid Quantification of Serum Ferritin].

    PubMed

    Shibusa, Kotoe; Hatayama, Mayumi; Toki, Yasumichi; Yamamoto, Masayo; Ito, Satoshi; Shindo, Motohiro; Fujiya, Mikihiro; Niizeki, Noriyasu; Tomoda, Yutaka; Kawai, Yuichi; Addo, Lynda; Ikuta, Katsuya

    2015-12-01

    Serum ferritin is an excellent marker for total iron content in the body and is essential for the diagnosis of iron deficiency or iron overload. Recently, a simple and rapid method, which utilizes immunochromatography for the quantification of serum ferritin, was developed. However, the range of measurement in previous reagents was limited (10-500 ng/mL). This range is rather narrow and is not fully helpful for the diagnosis of iron overload which sometimes occurs as a result of prolonged transfusions, or for monitoring iron contents during iron chelation therapy against iron overload. In the present study we evaluated the basic performance of the newly developed "Point Strip ferritin-3000", which can measure serum ferritin in the range of 300-3,000 ng/mL. Coefficient of variation (CV) s of within and inter-day assays were in the ranges of 7.3-11.1% and 2.1-5.2%, respectively. Using 87 serum samples obtained from the patients with written informed consents, the correlation coefficient was calculated to be 0.93 compared to the control method. In addition, the quantification of serum ferritin by "Point Strip ferritin-3000" was not influenced by bilirubin, hemoglobin, chyle, rheumatoid factor, or ascorbic acid. From our data, "Point Strip ferritin-3000" is reliable reagent in the range of 300-3,000 ng/mL, and is therefore considered to be useful for the diagnosis of iron overload, as well as for monitoring iron contents during iron chelation therapy. In addition, this quantification method can be easily performed using a small desktop equipment without any special technique, making this system applicable for epidemiological surveys and clinical studies. PMID:27089653

  16. Social Network and Risk-Taking Behavior Most Associated with Rapid HIV Testing, Circumcision, and Preexposure Prophylaxis Acceptability Among High-Risk Indian Men

    PubMed Central

    Kumar, Rupali; Dandona, Rakhi; Kumar, Prem; Kumar, Anil; Lakshmi, Vemu; Laumann, Edward; Mayer, Kenneth; Dandona, Lalit

    2012-01-01

    Abstract Indian truck drivers and their younger apprentice drivers are at increased risk of HIV infection. We determine network and risk practices associated with willingness to adopt HIV prevention interventions currently not being used in India: rapid HIV testing, circumcision, and preexposure prophylaxis (PrEP) in order to inform the National AIDS Control Program (NACP). Truck drivers and truck cleaners were systematically recruited to participate in a social network and risk survey in Hyderabad, Southern India. Three separate composite measures of acceptability of rapid HIV testing, circumcision, and PrEP acceptability were utilized to independently assess the relationship of these prevention interventions with risk-practices and social network characteristics. An 89% participation rate yielded 1602 truck drivers and truck cleaners with 54.2% younger than 30 years of age and 2.8% HIV infected. Twenty-five percent of respondents reported sex with female sex workers (FSW) and 5% with men (MSM). Rapid testing, circumcision, and PrEP acceptability were 97.4%, 9.1%, and 85.9%, respectively. Participants reporting prosocial network characteristics were more accepting of rapid testing (adjusted odds ratio [AORs] 3.07–6.71; p<0.05) and demonstrated variable PrEP acceptability (AORs 0.08–2.22; p<0.001). Sex with FSWs was associated with PrEP acceptability (AOR 4.27; p<0.001); sex with MSM was associated with circumcision acceptability only (AOR 2.66; p<0.01). Social network factors and risk-practices were associated with novel prevention acceptability, but not consistently across intervention type and with variable directionality. The NACP will need to consider that intervention uptake may likely be most successful when efforts are targeted to individuals with specific behavior and social network characteristics. PMID:22973951

  17. A SIMPLE AND RAPID MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY METHOD TO SCREEN FISH PLASMA SAMPLES FOR ESTROGEN-RESPONSIVE BIOMARKERS

    EPA Science Inventory

    In this study, we describe and evaluate the performance of a simple and rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers using matrix assisted laster desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) couopled with a short...

  18. A simple, rapid, and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A simple, rapid, and high-throughput fluorescent polarization immunoassay (FPIA) for simultaneous determination of organophosphorus pesticides (OPs) was developed. Three haptens were labeled with a fluorescein probe and used as tracers to develop a homogenous FPIA using a broad-specificity monoclon...

  19. Evaluation of Performance of Two Rapid Tests for Detection of HIV-1 and -2 in High- and Low-Prevalence Populations in Nigeria.

    PubMed

    Manak, Mark M; Njoku, Ogbonnaya S; Shutt, Ashley; Malia, Jennifer; Jagodzinski, Linda L; Milazzo, Mark; Suleiman, Aminu; Ogundeji, Amos A; Nelson, Robert; Ayemoba, Ojor R; O'Connell, Robert J; Singer, Darrell E; Michael, Nelson L; Peel, Sheila A

    2015-11-01

    The availability of reliable human immunodeficiency virus types 1 and 2 (HIV-1/2) rapid tests in resource-limited settings represents an important advancement in the accurate diagnosis of HIV infection and presents opportunities for implementation of effective prevention and treatment interventions among vulnerable populations. A study of the potential target populations for future HIV vaccine studies examined the prevalence of HIV infections at six selected sites in Nigeria and evaluated the use of two rapid diagnostic tests (RDTs) for HIV. The populations included market workers at sites adjacent to military installations and workers at highway settlements (truck stops) who may have a heightened risk of HIV exposure. Samples from 3,187 individuals who provided informed consent were tested in parallel using the Determine (DT) and Stat-Pak (SP) RDTs; discordant results were subjected to the Uni-Gold (UG) RDT as a tiebreaker. The results were compared to those of a third-generation enzyme immunoassay screen with confirmation of repeat reactive samples by HIV-1 Western blotting. One participant was HIV-2 infected, yielding positive results on both RDTs. Using the laboratory algorithm as a gold standard, we calculated sensitivities of 98.5% (confidence interval [CI], 97.1 to 99.8%) for DT and 98.1% (CI, 96.7 to 99.6%) for SP and specificities of 98.7% (CI, 98.3 -99.1%) for DT and 99.8% (CI, 99.6 to 100%) for SP. Similar results were obtained when the sites were stratified into those of higher HIV prevalence (9.4% to 22.8%) versus those of lower prevalence (3.2% to 7.3%). A parallel two-test algorithm requiring both DT and SP to be positive resulted in the highest sensitivity (98.1%; CI, 96.7 to 99.6%) and specificity (99.97%; CI, 99.9 to 100%) relative to those for the reference laboratory algorithm. PMID:26311857

  20. Will Gay and Bisexually Active Men at High Risk of Infection Use Over-the-Counter Rapid HIV Tests to Screen Sexual Partners?

    PubMed Central

    Carballo-Diéguez, Alex; Frasca, Timothy; Dolezal, Curtis; Balan, Ivan

    2013-01-01

    The Food and Drug Administration may license OraQuick™, a rapid HIV test, for over-the-counter (OTC) sale. We investigated whether HIV-uninfected, non-monogamous gay and bisexual men who never or rarely use condoms would use the test with partners as a harm-reduction approach. Sixty participants responded to two computer-assisted self-interviews, underwent an in-depth interview, and chose whether to test themselves with OraQuick™. Over 80% of the men said they would use the kit to test sexual partners or themselves if it became available OTC. Most participants understood that antibody tests have a window period in which the virus is undetectable yet saw advantages to using the test to screen partners; 74% tested themselves in our offices. Participants offered several possible strategies to introduce the home-test idea to partners, frequently endorsed mutual testing, and highlighted that home testing could stimulate greater honesty in serostatus disclosure. Participants drew distinctions between testing regular versus occasional partners. Non-monogamous MSM who never or rarely use condoms may nevertheless seek to avoid HIV. Technologies that do not interfere with sexual pleasure are likely to be used when available. Studies are needed to evaluate the advantages and disadvantages of using OTC rapid HIV tests as one additional harm-reduction tool. PMID:22293029

  1. Will gay and bisexually active men at high risk of infection use over-the-counter rapid HIV tests to screen sexual partners?

    PubMed

    Carballo-Diéguez, Alex; Frasca, Timothy; Dolezal, Curtis; Balan, Ivan

    2012-01-01

    The Food and Drug Administration may license OraQuick™, a rapid HIV test, for over-the-counter (OTC) sale. This study investigated whether HIV-uninfected, non-monogamous, gay and bisexual men who never or rarely use condoms would use the test with partners as a harm-reduction approach. Sixty participants responded to two computer-assisted self-interviews, underwent an in-depth interview, and chose whether to test themselves with OraQuick. Over 80% of the men said they would use the kit to test sexual partners or themselves if it became available OTC. Most participants understood that antibody tests have a window period in which the virus is undetectable, yet saw advantages to using the test to screen partners; 74% tested themselves in our offices. Participants offered several possible strategies to introduce the home-test idea to partners, frequently endorsed mutual testing, and highlighted that home testing could stimulate greater honesty in serostatus disclosure. Participants drew distinctions between testing regular versus occasional partners. Non-monogamous men who have sex with men, who never or rarely use condoms, may nevertheless seek to avoid HIV. Technologies that do not interfere with sexual pleasure are likely to be used when available. Studies are needed to evaluate the advantages and disadvantages of using OTC rapid HIV tests as one additional harm-reduction tool. PMID:22293029

  2. Non-instrumented incubation of a recombinase polymerase amplification assay for the rapid and sensitive detection of proviral HIV-1 DNA.

    PubMed

    Lillis, Lorraine; Lehman, Dara; Singhal, Mitra C; Cantera, Jason; Singleton, Jered; Labarre, Paul; Toyama, Anthony; Piepenburg, Olaf; Parker, Mathew; Wood, Robert; Overbaugh, Julie; Boyle, David S

    2014-01-01

    Sensitive diagnostic tests for infectious diseases often employ nucleic acid amplification technologies (NAATs). However, most NAAT assays, including many isothermal amplification methods, require power-dependent instrumentation for incubation. For use in low resource settings (LRS), diagnostics that do not require consistent electricity supply would be ideal. Recombinase polymerase amplification (RPA) is an isothermal amplification technology that has been shown to typically work at temperatures ranging from 25-43°C, and does not require a stringent incubation temperature for optimal performance. Here we evaluate the ability to incubate an HIV-1 RPA assay, intended for use as an infant HIV diagnostic in LRS, at ambient temperatures or with a simple non-instrumented heat source. To determine the range of expected ambient temperatures in settings where an HIV-1 infant diagnostic would be of most use, a dataset of the seasonal range of daily temperatures in sub Saharan Africa was analyzed and revealed ambient temperatures as low as 10°C and rarely above 43°C. All 24 of 24 (100%) HIV-1 RPA reactions amplified when incubated for 20 minutes between 31°C and 43°C. The amplification from the HIV-1 RPA assay under investigation at temperatures was less consistent below 30°C. Thus, we developed a chemical heater to incubate HIV-1 RPA assays when ambient temperatures are between 10°C and 30°C. All 12/12 (100%) reactions amplified with chemical heat incubation from ambient temperatures of 15°C, 20°C, 25°C and 30°C. We also observed that incubation at 30 minutes improved assay performance at lower temperatures where detection was sporadic using 20 minutes incubation. We have demonstrated that incubation of the RPA HIV-1 assay via ambient temperatures or using chemical heaters yields similar results to using electrically powered devices. We propose that this RPA HIV-1 assay may not need dedicated equipment to be a highly sensitive tool to diagnose infant HIV-1 in

  3. Elevated Basal Pre-infection CXCL10 in Plasma and in the Small Intestine after Infection Are Associated with More Rapid HIV/SIV Disease Onset

    PubMed Central

    Ploquin, Mickaël J.; Casrouge, Armanda; Huot, Nicolas; Passaes, Caroline; Lécuroux, Camille; Essat, Asma; Boufassa, Faroudy; Jacquelin, Béatrice; Jochems, Simon P.; Petitjean, Gaël; Angin, Mathieu; Gärtner, Kathleen; Garcia-Tellez, Thalía; Booiman, Thijs; Boeser-Nunnink, Brigitte D.; Roques, Pierre; Saez-Cirion, Asier; Vaslin, Bruno; Dereudre-Bosquet, Nathalie; Barré-Sinoussi, Françoise; Ghislain, Mathilde; Rouzioux, Christine; Lambotte, Olivier; Albert, Matthew L.; Goujard, Cécile; Kootstra, Neeltje; Meyer, Laurence; Müller-Trutwin, Michaela C.

    2016-01-01

    Elevated blood CXCL10/IP-10 levels during primary HIV-1 infection (PHI) were described as an independent marker of rapid disease onset, more robust than peak viremia or CD4 cell nadir. IP-10 enhances the recruitment of CXCR3+ cells, which include major HIV-target cells, raising the question if it promotes the establishment of viral reservoirs. We analyzed data from four cohorts of HIV+ patients, allowing us to study IP-10 levels before infection (Amsterdam cohort), as well as during controlled and uncontrolled viremia (ANRS cohorts). We also addressed IP-10 expression levels with regards to lymphoid tissues (LT) and blood viral reservoirs in patients and non-human primates. Pre-existing elevated IP-10 levels but not sCD63 associated with rapid CD4 T-cell loss upon HIV-1 infection. During PHI, IP-10 levels and to a lesser level IL-18 correlated with cell-associated HIV DNA, while 26 other inflammatory soluble markers did not. IP-10 levels tended to differ between HIV controllers with detectable and undetectable viremia. IP-10 was increased in SIV-exposed aviremic macaques with detectable SIV DNA in tissues. IP-10 mRNA was produced at higher levels in the small intestine than in colon or rectum. Jejunal IP-10+ cells corresponded to numerous small and round CD68neg cells as well as to macrophages. Blood IP-10 response negatively correlated with RORC (Th17 marker) gene expression in the small intestine. CXCR3 expression was higher on memory CD4+ T cells than any other immune cells. CD4 T cells from chronically infected animals expressed extremely high levels of intra-cellular CXCR3 suggesting internalization after ligand recognition. Elevated systemic IP-10 levels before infection associated with rapid disease progression. Systemic IP-10 during PHI correlated with HIV DNA. IP-10 production was regionalized in the intestine during early SIV infection and CD68+ and CD68neg haematopoietic cells in the small intestine appeared to be the major source of IP-10. PMID:27509048

  4. Cost-Effectiveness of Pooled Nucleic Acid Amplification Testing for Acute HIV Infection after Third-Generation HIV Antibody Screening and Rapid Testing in the United States: A Comparison of Three Public Health Settings

    PubMed Central

    Hutchinson, Angela B.; Patel, Pragna; Sansom, Stephanie L.; Farnham, Paul G.; Sullivan, Timothy J.; Bennett, Berry; Kerndt, Peter R.; Bolan, Robert K.; Heffelfinger, James D.; Prabhu, Vimalanand S.; Branson, Bernard M.

    2010-01-01

    Background Detection of acute HIV infection (AHI) with pooled nucleic acid amplification testing (NAAT) following HIV testing is feasible. However, cost-effectiveness analyses to guide policy around AHI screening are lacking; particularly after more sensitive third-generation antibody screening and rapid testing. Methods and Findings We conducted a cost-effectiveness analysis of pooled NAAT screening that assessed the prevention benefits of identification and notification of persons with AHI and cases averted compared with repeat antibody testing at different intervals. Effectiveness data were derived from a Centers for Disease Control and Prevention AHI study conducted in three settings: municipal sexually transmitted disease (STD) clinics, a community clinic serving a population of men who have sex with men, and HIV counseling and testing sites. Our analysis included a micro-costing study of NAAT and a mathematical model of HIV transmission. Cost-effectiveness ratios are reported as costs per quality-adjusted life year (QALY) gained in US dollars from the societal perspective. Sensitivity analyses were conducted on key variables, including AHI positivity rates, antibody testing frequency, symptomatic detection of AHI, and costs. Pooled NAAT for AHI screening following annual antibody testing had cost-effectiveness ratios exceeding US$200,000 per QALY gained for the municipal STD clinics and HIV counseling and testing sites and was cost saving for the community clinic. Cost-effectiveness ratios increased substantially if the antibody testing interval decreased to every 6 months and decreased to cost-saving if the testing interval increased to every 5 years. NAAT was cost saving in the community clinic in all situations. Results were particularly sensitive to AHI screening yield. Conclusions Pooled NAAT screening for AHI following negative third-generation antibody or rapid tests is not cost-effective at recommended antibody testing intervals for high-risk persons

  5. Rapid HIV Testing Is Highly Acceptable and Preferred among High-Risk Gay And Bisexual Men after Implementation in Sydney Sexual Health Clinics

    PubMed Central

    Conway, Damian P.; Guy, Rebecca; Davies, Stephen C; Couldwell, Deborah L.; McNulty, Anna; Smith, Don E.; Keen, Phillip; Cunningham, Philip; Holt, Martin

    2015-01-01

    Background Rapid HIV testing (RHT) is well established in many countries, but it is new in Australia. We assessed the acceptability of RHT and its associations among gay, bisexual and other men who have sex with men (GBM) after implementation of RHT in Sydney sexual health clinics. Methods GBM were invited to complete an acceptability questionnaire before and after provision of the result of finger-prick blood RHT, comparing their experience of RHT with conventional HIV testing (CHT) involving venipuncture. Logistic regression was used to assess associations between patient characteristics and the preference for RHT over CHT next time they tested for HIV. Results Of 1061 GBM who received non-reactive RHT results, 59% found RHT less stressful than CHT and 34% reported no difference, and 61% found RHT more comfortable than CHT and 26% reported no difference. Nearly all men were satisfied with RHT result delivery (99%) and the RHT process overall (99%). Most men (79%) preferred RHT for their next HIV test and this preference was stronger in men who were aged 35-44 years (adjusted odds ratio [AOR] 2.49, p<0.01), reported they would test more often if RHT was available (AOR 1.66, p=0.01), found returning for results annoying (AOR 1.67, p=0.01), and found RHT less stressful (AOR 2.37, p<0.01) and more comfortable (AOR 1.62, p=0.02) than CHT. Men concerned about the reliability of RHT were less than half as likely to prefer RHT for their next HIV test (AOR 0.44, p<0.01). Conclusions Most GBM preferred RHT to CHT next time and this preference was associated with finding RHT more convenient, more comfortable and less stressful than CHT. These findings suggest that in a clinic setting RHT should be considered to improve the patient experience and may potentially increase uptake and frequency of HIV testing. PMID:25898140

  6. HIV treatment as prevention: how scientific discovery occurred and translated rapidly into policy for the global response.

    PubMed

    Cohen, Myron S; Holmes, Charles; Padian, Nancy; Wolf, Megan; Hirnschall, Gottfried; Lo, Ying-Ru; Goosby, Eric

    2012-07-01

    In 2011 interim results of HIV Prevention Trials Network study 052, a National Institutes of Health study designed to test the effectiveness of antiretroviral treatment against the spread of HIV, were reported. These results showed that in a stable relationship in which one member of the couple was infected with HIV, treatment of the infected partner with antiretroviral drugs, combined with couples counseling and condom use, resulted in a 96 percent reduction in sexual transmission of HIV-1. This finding led to the use of antiretroviral treatment as a cornerstone of HIV prevention. Independent advisory committees of the President's Emergency Plan for AIDS Relief (PEPFAR) and the World Health Organization (WHO) have since issued analyses that set the stage for broader use of antiretroviral agents in treatment and prevention. This article describes the separate PEPFAR and WHO recommendations and outlines the design of prospective new trials to test how best to maximize the benefits of early treatment for prevention. PMID:22778333

  7. Assessment of Overlap of Phylogenetic Transmission Clusters and Communities in Simple Sexual Contact Networks: Applications to HIV-1

    PubMed Central

    Villandre, Luc; Günthard, Huldrych F.; Kouyos, Roger; Stadler, Tanja

    2016-01-01

    Background Transmission patterns of sexually-transmitted infections (STIs) could relate to the structure of the underlying sexual contact network, whose features are therefore of interest to clinicians. Conventionally, we represent sexual contacts in a population with a graph, that can reveal the existence of communities. Phylogenetic methods help infer the history of an epidemic and incidentally, may help detecting communities. In particular, phylogenetic analyses of HIV-1 epidemics among men who have sex with men (MSM) have revealed the existence of large transmission clusters, possibly resulting from within-community transmissions. Past studies have explored the association between contact networks and phylogenies, including transmission clusters, producing conflicting conclusions about whether network features significantly affect observed transmission history. As far as we know however, none of them thoroughly investigated the role of communities, defined with respect to the network graph, in the observation of clusters. Methods The present study investigates, through simulations, community detection from phylogenies. We simulate a large number of epidemics over both unweighted and weighted, undirected random interconnected-islands networks, with islands corresponding to communities. We use weighting to modulate distance between islands. We translate each epidemic into a phylogeny, that lets us partition our samples of infected subjects into transmission clusters, based on several common definitions from the literature. We measure similarity between subjects’ island membership indices and transmission cluster membership indices with the adjusted Rand index. Results and Conclusion Analyses reveal modest mean correspondence between communities in graphs and phylogenetic transmission clusters. We conclude that common methods often have limited success in detecting contact network communities from phylogenies. The rarely-fulfilled requirement that network

  8. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy

    PubMed Central

    Tyson, Adam L.; Hilton, Stephen T.; Andreae, Laura C.

    2015-01-01

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  9. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy.

    PubMed

    Tyson, Adam L; Hilton, Stephen T; Andreae, Laura C

    2015-10-30

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  10. Poor performance of the determine HIV-1/2 Ag/Ab combo fourth-generation rapid test for detection of acute infections in a National Household Survey in Swaziland.

    PubMed

    Duong, Yen T; Mavengere, Yvonne; Patel, Hetal; Moore, Carole; Manjengwa, Julius; Sibandze, Dumile; Rasberry, Christopher; Mlambo, Charmaine; Li, Zhi; Emel, Lynda; Bock, Naomi; Moore, Jan; Nkambule, Rejoice; Justman, Jessica; Reed, Jason; Bicego, George; Ellenberger, Dennis L; Nkengasong, John N; Parekh, Bharat S

    2014-10-01

    Fourth-generation HIV rapid tests (RTs) claim to detect both p24 antigen (Ag) and HIV antibodies (Ab) for early identification of acute infections, important for targeting prevention and reducing HIV transmission. In a nationally representative household survey in Swaziland, 18,172 adults, age 18 to 49 years, received home-based HIV rapid testing in 2010 and 2011. Of the 18,172 individuals, 5,822 (32.0%) were Ab positive (Ab(+)) by the Determine HIV-1/2 Ab/Ab combo test, and 5,789 (99.4%) of those were confirmed to be reactive in the Uni-Gold test. Determine combo identified 12 individuals as having acute infections (Ag(+)/Ab negative [Ab(-)]); however, none had detectable HIV-1 RNA and 8 of 12 remained HIV negative at their 6-week follow-up visit (4 were lost to follow-up). All RT-nonreactive samples were pooled and tested by nucleic acid amplification testing (NAAT) to identify acute infections. NAAT identified 13 (0.1%) of the 12,338 HIV antibody-negative specimens as HIV RNA positive, with RNA levels ranging from 300 to >10,000,000 copies/ml. However, none of them were Ag(+) by Determine combo. Follow-up testing of 12 of the 13 NAAT-positive individuals at 6 months demonstrated 12 seroconversions (1 individual was lost to follow-up). Therefore, the Determine combo test had a sensitivity of 0% (95% confidence interval, 0 to 28) and positive predictive value of 0% for the detection of acute infections. The ability of the 4th-generation Determine combo to detect antigen was very poor in Swaziland. Thus, the Determine combo test does not add any value to the current testing algorithm; rather, it adds additional costs and complexity to HIV diagnosis. The detection of acute HIV infections may need to rely on other testing strategies. PMID:25122853

  11. A simple and rapid creatinine sensing via DLS selectivity, using calix[4]arene thiol functionalized gold nanoparticles.

    PubMed

    Sutariya, Pinkesh G; Pandya, Alok; Lodha, Anand; Menon, Shobhana K

    2016-01-15

    A new, simple, ultra-sensitive and selective approach has been reported for the "on spot" colorimetric detection of creatinine based on calix[4]arene functionalized gold nanoparticles (AuNPs) with excellent discrimination in the presence of other biomolecules. The lower detection limit of the method is 2.16nM. The gold nanoparticles and p-tert-butylcalix[4]arene were synthesized by microwave assisted method. Specifically, in our study, we used dynamic light scattering (DLS) which is a powerful method for the determination of small changes in particle size, improved selectivity and sensitivity of the creatinine detection system over colorimetric method. The nanoassembly is characterized by Transmission electron microscopy (TEM), DLS, UV-vis and ESI-MS spectroscopy, which demonstrates the binding affinity due its ability of hydrogen bonding and electrostatic interaction between -NH group of creatinine and pSDSC4. It exhibits fast response time (<60s) to creatinine and has long shelf-life (>5 weeks). The developed pSDSC4-AuNPs based creatinine biosensor will be established as simple, reliable and accurate tool for the determination of creatinine in human urine samples. PMID:26592650

  12. Potential application of immunoassays for simple, rapid and quantitative detections of phytoavailable neonicotinoid insecticides in cropland soils.

    PubMed

    Watanabe, Eiki; Seike, Nobuyasu; Motoki, Yutaka; Inao, Keiya; Otani, Takashi

    2016-10-01

    This study evaluated the applicability of commercially available kit-based enzyme-linked immunosorbent assay (ELISA) to simple, quick, and quantitative detection for three water-extractable (phytoavailable) neonicotinoid insecticides: dinotefuran, clothianidin, and imidacloprid in soils. ELISA showed excellent analytical sensitivity for determination, but with cross-reaction to structurally related neonicotinoid analogues, which might produce false positives. To analyze insecticides in soil samples of diverse physicochemical properties, they were extracted with water. The aqueous soil extracts were assayed directly with ELISA. No matrix interference was observed without additional dilution with water. Recovery experiments for the insecticides from aqueous soil extracts spiked at 2-10 ng/mL showed good accuracy (72-126%) and precision (<16%). Kit-based ELISAs were used to estimate soil-water distribution coefficients (Kd). Values estimated using this method showed positive correlation between organic carbon contents in soil and those for evaluated insecticides. Results indicate that the evaluated kit-based ELISA has applicability for simple, quick, and reliable detection of phytoavailable insecticides in soils and for estimating Kd values in soil. PMID:27344017

  13. A simple and rapid one-time method to evaluate the non-acidic gas content from bioprocesses.

    PubMed

    Bassard, David; André, Laura; Dotal, Nicolas; Valentin, Ludovic; Nonus, Maurice; Pauss, André; Ribeiro, Thierry

    2014-02-01

    This paper presents a rapid less than 2 min and low-cost method involving the use of alkali solution to capture the acidic gasses from a biogas, thereby providing an estimate of the percentage of non-acidic gasses. Such a method was mentioned in the literature but never fully described or optimized. After sampling an aliquot of gas from bioprocess, gas was injected in a sealed flask with a 3 M NaOH solution, and after equilibrium was obtained, the non-acidic gas volume was measured. The method was first calibrated with certified gasses with an accuracy observed between 98 and 105%. Regarding the validation step, certified standard gas mixtures and nine biogas-laboratory batch reactors were used, the overall accuracy reported was 103 + 3%. This rapid and low-cost method may either be used in laboratory conditions as a quick and low cost alternative to standard analysis equipment or in addition as a routine field control method used on full-scale plants. PMID:23743732

  14. A simple and rapid detection of tissue adhesive-induced biochemical changes in cells and DNA using Raman spectroscopy

    PubMed Central

    Jung, Gyeong Bok; Lee, Young Ju; Lee, Gihyun; Park, Hun-Kuk

    2013-01-01

    We demonstrate a cytotoxicity evaluation of tissue adhesive using Raman spectroscopy. This method allows for quantitative, label-free, non-invasive and rapid monitoring of the biochemical changes of cells following tissue adhesive treatment. Here, we show the biochemical property changes in mouse fibroblast L929 cells and cellular DNA following tissue adhesive (Dermabond) treatment using Raman spectroscopy. The Raman band intensities were significantly decreased when the cells were treated with Dermabond as compared to control cells. These results suggest denaturation and conformational changes in proteins and degradation of DNA related to cell death. To support these conclusions, conventional cytotoxicity assays such as WST, LIVE/DEAD, and TUNEL were carried out, and the results were in agreement with the Raman results. Thus, Raman spectroscopy analysis not only distinguishes between viable and damaged cells, but can also be used for identification and quantification of a cytotoxicity of tissue adhesive, which based on the cellular biochemical and structural changes at a molecular level. Therefore, we suggest that this method could be used for cytotoxic evaluation of tissue adhesives by rapid and sensitive detection of cellular changes. PMID:24298425

  15. Development of a Latex Agglutination Test as a Simple and Rapid Method for Diagnosis of Trichomonas vaginalis Infection.

    PubMed

    Darani, Hossein Yousofi; Ahmadi, Firuzeh; Zabardast, Nozhat; Yousefi, Hossein Ali; Shirzad, Hedayat

    2010-01-01

    Trichomoniasis is a worldwide infection and due to its complications rapid and accurate diagnosis of infection especially in pregnant women is very important. In this study, development of a latex agglutination test using native antigens for rapid diagnosis of trichomoniasis is investigated. Trichomonas vaginalis was harvested from TYIS33 culture medium and anti Trichomonas vaginalis antiserum was raised in rabbits. Salt precipitation method was used for antibody purification. Polyesteren latex particles coated with purified antibody and used for detection of Trichomonas vaginalis. Clinical samples of vaginal discharge were collected from 500 women and examined for Trichomonas vaginalis by using wet mount, culture and latex agglutination tests. Sensitivity and specificity of latex test was determined considering culture as golden standard. Sensitivity and specificity of latex agglutination test was 100% and 81% and those of wet mount were 33.3% and 100%, respectively. Positive and negative predictive values of latex agglutination test were 6% and 100%, respectively. Due to inconvenient sensitivity and specificity of the latex agglutination test developed in this study, further work is recommended to improve the test. PMID:23408769

  16. Processes and outcomes of training on rapid assessment and response methods on injecting drug use and related HIV infection in the Russian Federation.

    PubMed

    Burrows; Trautmann; Frost; Bijl; Sarankov; Sarang; Chernenko

    2000-03-01

    In September 1997, Médecins Sans Frontières-Holland (MSF-H) began a project to provide training and support for HIV/AIDS prevention among injecting drug users (IDUs) in the Russian Federation, focusing on the use of the World Health Organization Rapid Assessment and Response Guide on Injecting Drug Use, and the European Peer Support Manual. As part of the training, participants are asked to carry out a rapid situation assessment (RSA) in their city or region as a major step towards designing and implementing an effective program to prevent HIV transmission among IDUs. This paper focuses on the first four training cycles of the programme (from January 1998 to January 1999), in which 89 health professionals and others from 32 Russian cities took part. A total of 28 rapid situation assessments were completed or almost completed by participants during these four cycles. The paper provides an overview of the methods used and major problems faced by participants undertaking these assessments, as well as summarising the 14 harm reduction programmes which resulted. PMID:10699552

  17. Simple, rapid and reliable preparation of [¹¹C]-(+)-α-DTBZ of high quality for routine applications.

    PubMed

    Zhang, Jinming; Zhang, Xiaojun; Li, Yungang; Tian, Jiahe

    2012-01-01

    [¹¹C]-(+)-α-DTBZ has been used as a marker of dopaminergic terminal densities in human striatum and expressed in islet beta cells in the pancreas. We aimed to establish a fully automated and simple procedure for the synthesis of [¹¹C]-(+)-α-DTBZ for routine applications. [¹¹C]-(+)-α-DTBZ was synthesized from a 9-hydroxy precursor in acetone and potassium hydroxide with [¹¹C]-methyl triflate and was purified by solid phase extraction using a Vac tC-18 cartridge. Radiochemical yields based on [¹¹C]-methyl triflate (corrected for decay) were 82.3% ± 3.6%, with a specific radioactivity of 60 GBq/μmol. Time elapsed was less than 20 min from end of bombardment to release of the product for quality control. PMID:22728363

  18. Gaseous VOCs rapidly modify particulate matter and its biological effects - Part 1: Simple VOCs and model PM

    NASA Astrophysics Data System (ADS)

    Ebersviller, S.; Lichtveld, K.; Sexton, K. G.; Zavala, J.; Lin, Y.-H.; Jaspers, I.; Jeffries, H. E.

    2012-12-01

    This is the first of a three-part study designed to demonstrate dynamic entanglements among gaseous organic compounds (VOC), particulate matter (PM), and their subsequent potential biological effects. We study these entanglements in increasingly complex VOC and PM mixtures in urban-like conditions in a large outdoor chamber. To the traditional chemical and physical characterizations of gas and PM, we added new measurements of biological effects, using cultured human lung cells as model indicators. These biological effects are assessed here as increases in cellular damage or expressed irritation (i.e., cellular toxic effects) from cells exposed to chamber air relative to cells exposed to clean air. The exposure systems permit virtually gas-only- or PM-only-exposures from the same air stream containing both gases and PM in equilibria, i.e., there are no extractive operations prior to cell exposure. Our simple experiments in this part of the study were designed to eliminate many competing atmospheric processes to reduce ambiguity in our results. Simple volatile and semi-volatile organic gases that have inherent cellular toxic properties were tested individually for biological effect in the dark (at constant humidity). Airborne mixtures were then created with each compound to which we added PM that has no inherent cellular toxic properties for another cellular exposure. Acrolein and p-tolualdehyde were used as model VOCs and mineral oil aerosol (MOA) was selected as a surrogate for organic-containing PM. MOA is appropriately complex in composition to represent ambient PM, and exhibits no inherent cellular toxic effects and thus did not contribute any biological detrimental effects on its own. Chemical measurements, combined with the responses of our biological exposures, clearly demonstrate that gas-phase pollutants can modify the composition of PM (and its resulting detrimental effects on lung cells). We observed that, even if the gas-phase pollutants are not

  19. Gaseous VOCs rapidly modify particulate matter and its biological effects - Part 1: Simple VOCs and model PM

    NASA Astrophysics Data System (ADS)

    Ebersviller, S.; Lichtveld, K.; Sexton, K. G.; Zavala, J.; Lin, Y.-H.; Jaspers, I.; Jeffries, H. E.

    2012-02-01

    This is the first of a three-part study designed to demonstrate dynamic entanglements among gaseous organic compounds (VOC), particulate matter (PM), and their subsequent potential biological effects. We study these entanglements in increasingly complex VOC and PM mixtures in urban-like conditions in a large outdoor chamber. To the traditional chemical and physical characterizations of gas and PM, we added new measurements of gas-only- and PM-only-biological effects, using cultured human lung cells as model indicators. These biological effects are assessed here as increases in cellular damage or expressed irritation (i.e., cellular toxic effects) from cells exposed to chamber air relative to cells exposed to clean air. The exposure systems permit gas-only- or PM-only-exposures from the same air stream containing both gases and PM in equilibria, i.e., there are no extractive operations prior to cell exposure. Our simple experiments in this part of the study were designed to eliminate many competing atmospheric processes to reduce ambiguity in our results. Simple volatile and semi-volatile organic gases that have inherent cellular toxic properties were tested individually for biological effect in the dark (at constant humidity). Airborne mixtures were then created with each compound and PM that has no inherent cellular toxic properties for another cellular exposure. Acrolein and p-tolualdehyde were used as model VOCs and mineral oil aerosol (MOA) was selected as a surrogate for organic-containing PM. MOA is appropriately complex in composition to represent ambient PM, and it exhibits no inherent cellular toxic effects and thus did not contribute any biological detrimental effects on its own. Chemical measurements, combined with the responses of our biological exposures, clearly demonstrate that gas-phase pollutants can modify the composition of PM (and its resulting detrimental effects on lung cells) - even if the gas-phase pollutants are not considered likely to

  20. AFSM sequencing approach: a simple and rapid method for genome-wide SNP and methylation site discovery and genetic mapping

    PubMed Central

    Xia, Zhiqiang; Zou, Meiling; Zhang, Shengkui; Feng, Binxiao; Wang, Wenquan

    2014-01-01

    We describe methods for the assessment of amplified-fragment single nucleotide polymorphism and methylation (AFSM) sites using a quick and simple molecular marker-assisted breeding strategy based on the use of two restriction enzyme pairs (EcoRI-MspI and EcoRI-HpaII) and a next-generation sequencing platform. Two sets of 85 adapter pairs were developed to concurrently identify SNPs, indels and methylation sites for 85 lines of cassava population in this study. In addition to SNPs and indels, the simplicity of the AFSM protocol makes it particularly suitable for high-throughput full methylation and hemi-methylation analyses. To further demonstrate the ease of this approach, a cassava genetic linkage map was constructed. This approach should be widely applicable for genetic mapping in a variety of organisms and will improve the application of crop genomics in assisted breeding. PMID:25466435

  1. Development and Evaluation of Two Simple, Rapid Immunochromatographic Tests for the Detection of Yersinia pestis Antibodies in Humans and Reservoirs

    PubMed Central

    Rajerison, Minoarisoa; Dartevelle, Sylvie; Ralafiarisoa, Lalao A.; Bitam, Idir; Tuyet, Dinh Thi Ngoc; Andrianaivoarimanana, Voahangy; Nato, Faridabano; Rahalison, Lila

    2009-01-01

    Background Tools for plague diagnosis and surveillance are not always available and affordable in most of the countries affected by the disease. Yersinia pestis isolation for confirmation is time-consuming and difficult to perform under field conditions. Serologic tests like ELISA require specific equipments not always available in developing countries. In addition to the existing rapid test for antigen detection, a rapid serodiagnostic assay may be useful for plague control. Methods/Principal Findings We developed two rapid immunochromatography-based tests for the detection of antibodies directed against F1 antigen of Y. pestis. The first test, SIgT, which detects total Ig (IgT) anti-F1 in several species (S) (human and reservoirs), was developed in order to have for the field use an alternative method to ELISA. The performance of the SIgT test was evaluated with samples from humans and animals for which ELISA was used to determine the presumptive diagnosis of plague. SIgT test detected anti-F1 Ig antibodies in humans with a sensitivity of 84.6% (95% CI: 0.76–0.94) and a specificity of 98% (95% CI: 0.96–1). In evaluation of samples from rodents and other small mammals, the SlgT test had a sensitivity of 87.8% (95% CI: 0.80–0.94) and a specificity of 90.3% (95% CI: 0.86–0.93). Improved performance was obtained with samples from dogs, a sentinel animal, with a sensitivity of 93% (95% CI: 0.82–1) and a specificity of 98% (95% CI: 0.95–1.01). The second test, HIgM, which detects human (H) IgM anti-F1, was developed in order to have another method for plague diagnosis. Its sensitivity was 83% (95% CI: 0.75–0.90) and its specificity about 100%. Conclusion/Significance The SIgT test is of importance for surveillance because it can detect Ig antibodies in a range of reservoir species. The HIgM test could facilitate the diagnosis of plague during outbreaks, particularly when only a single serum sample is available. PMID:19399164

  2. Responsiveness of a simple RAPID-3-like index compared to disease-specific BASDAI and ASDAS indices in patients with axial spondyloarthritis

    PubMed Central

    Castrejón, Isabel; Pincus, Theodore; Wendling, Daniel; Dougados, Maxime

    2016-01-01

    Objective To evaluate the responsiveness of a simple routine assessment of patient index data (RAPID3)-like index that includes only 3 patient self-report measures (physical function, pain and patient global estimate) compared to that of traditional composite indices to assess change in patients with axial spondyloarthritis (Ax-SpA). Methods Devenir des Spondylarthropathies Indifférenciées Récentes (DESIR) is a prospective cohort of patients with inflammatory back pain suggestive of Ax-SpA. The study included 461 patients, who met the Assessment of SpondyloArthritis International Society (ASAS) classification criteria for Ax-SpA. A simple RAPID3-like index was compared with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and the AS Disease Activity Score (ASDAS) scores for responsiveness over 6 months. Construct validity was also evaluated through Pearson correlations and discrimination of disease activity through standardised mean differences for the 3 indices. Results The RAPID3-like index was correlated significantly with BASDAI (r=0.84, p<0.005) and ASDAS-C-reactive protein (CRP) (r=0.74, p<0.005), similar to correlations of BASDAI with ASDAS-CRP (r=0.76, p<0.005). The percentage of patients with inactive disease ranged from 9% to 25% and with high activity from 10% to 45%, according to various measures. The capacity to discriminate between high and low disease activity was similar for the 3 indices. The strength of agreement of RAPID3 with ASDAS-CRP was moderate (0.44) and lower with BASDAI (0.37). Responsiveness over 6 months was slightly higher for ASDAS-CRP and the RAPID3-like index than that for BASDAI. Conclusions The RAPID3-like index provides similar information to BASDAI and ASDAS-CRP concerning responsiveness over 6 months. RAPID3 appears feasible to assess patients with Ax-SpA quantitatively over time in busy clinical settings. PMID:27486525

  3. A simple, rapid, low-cost technique for naked-eye detection of urine-isolated TMPRSS2:ERG gene fusion RNA.

    PubMed

    Koo, Kevin M; Wee, Eugene J H; Mainwaring, Paul N; Trau, Matt

    2016-01-01

    The TMPRSS2:ERG gene fusion is one of a series of highly promising prostate cancer (PCa) biomarker alternatives to the controversial serum PSA. Current methods for detecting TMPRSS2:ERG are limited in terms of long processing time, high cost and the need for specialized equipment. Thus, there is an unmet need for less complex, faster, and cheaper methods to enable gene fusion detection in the clinic. We describe herein a simple, rapid and inexpensive assay which combines robust isothermal amplification technique with a novel visualization method for evaluating urinary TMPRSS2:ERG status at less than USD 5 and with minimal equipment. The assay is sensitive, and rapidly detects as low as 10(5) copies of TMPRSS2:ERG transcripts while maintaining high levels of specificity. PMID:27470540

  4. A simple, rapid, low-cost technique for naked-eye detection of urine-isolated TMPRSS2:ERG gene fusion RNA

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J. H.; Mainwaring, Paul N.; Trau, Matt

    2016-01-01

    The TMPRSS2:ERG gene fusion is one of a series of highly promising prostate cancer (PCa) biomarker alternatives to the controversial serum PSA. Current methods for detecting TMPRSS2:ERG are limited in terms of long processing time, high cost and the need for specialized equipment. Thus, there is an unmet need for less complex, faster, and cheaper methods to enable gene fusion detection in the clinic. We describe herein a simple, rapid and inexpensive assay which combines robust isothermal amplification technique with a novel visualization method for evaluating urinary TMPRSS2:ERG status at less than USD 5 and with minimal equipment. The assay is sensitive, and rapidly detects as low as 105 copies of TMPRSS2:ERG transcripts while maintaining high levels of specificity. PMID:27470540

  5. Exclusion and diagnosis of deep vein thrombosis by a rapid ELISA D-dimer test, compression ultrasonography, and a simple clinical model.

    PubMed

    Michiels, J J; Oortwijn, W J; Naaborg, R

    1999-07-01

    The classical clinical signs of deep vein thrombosis (DVT) are unspecific and may be found in several other conditions besides DVT. Therefore, patients suspicious of DVT are subjected to elaborate invasive or noninvasive evidence-based procedures that actually confirm DVT in only 20% to 30% of patients in this setting. However, simple laboratory tests and noninvasive strategies to exclude and diagnose DVT are becoming available in the clinical emergency setting of outpatients. In the presented literature, a sound basis is provided for quantifying clinical judgment for the diagnosis of acute proximal DVT. The number of positive clinical findings at time of first suspicion of DVT appears to correlate directly with the probability of acute proximal DVT. The modified clinical model of Landefeld and Wells for DVT allows reasonable accurate classification of patients into low, moderate, and high probability for suffering DVT. The rapid automated enzyme-linked immunoabsorbant assay (ELISA) VIDAS D-dimer presently available can be rapidly performed in daily practice and emergency situations and is accurate to a high degree, especially in ruling out ongoing venous thromboembolic processes. The sequential use of the rapid ELISA VIDAS D-dimer test and compression ultrasonography in a well-designed clinical setting using a simple clinical model predicts a significant improvement due to a high sensitivity near 100% for the exclusion and diagnosis of DVT in the majority of outpatients with suspect DVT. A prospective decision analysis management study is proposed to exclude and diagnose DVT based on the rapid ELISA VIDAS D-dimer test and compression ultrasonography within the context of a ready-to-use simple clinical model. The proposed simple model of a rational diagnosis of deep vein thrombosis (RADIA DVT) has to be tested in a large multicenter study of more than 1,000 outpatients with suspected DVT. This model would be less expensive, easy to perform, and likely yield a

  6. A microscale approach for simple and rapid monitoring of cell growth and lipid accumulation in Neochloris oleoabundans.

    PubMed

    Kwak, Ho Seok; Kim, Jaoon Young Hwan; Sim, Sang Jun

    2015-10-01

    Due to the increasing environmental problems caused by the use of fossil fuels, microalgae have been spotlighted as renewable resources to produce biomass and biofuels. Therefore, the investigation of the optimum culture conditions of microalgae in a short time is one of the important factors for improving growth and lipid productivity. Herein, we developed a PDMS-based high-throughput screening system to rapidly and easily determine the optimum conditions for high-density culture and lipid accumulation of Neochloris oleoabundans. Using the microreactor, we were able to find the optimal culture conditions of N. oleoabundans within 5 days by rapid and parallel monitoring growth and lipid induction under diverse conditions of light intensity, pH, CO2 and nitrate concentration. We found that the maximum growth rate (µ max = 2.13 day(-1)) achieved in the microreactor was 1.58-fold higher than that in a flask (µ max = 1.34 day(-1)) at the light intensity of 40 µmol photons m(-2) s(-1), 5 % CO2 (v/v), pH 7.5 and 7 mM nitrate. In addition, we observed that the accumulation of lipid in the microreactor was 1.5-fold faster than in a flask under optimum culture condition. These results show that the microscale approach has the great potential for improving growth and lipid productivity by high-throughput screening of diverse optimum conditions. PMID:26209175

  7. Diagnostic accuracy of the genotype MTBDRsl assay for rapid diagnosis of extensively drug-resistant tuberculosis in HIV-coinfected patients.

    PubMed

    Kontsevaya, Irina; Ignatyeva, Olga; Nikolayevskyy, Vladyslav; Balabanova, Yanina; Kovalyov, Alexander; Kritsky, Andrey; Matskevich, Olesya; Drobniewski, Francis

    2013-01-01

    The Russian Federation is a high-tuberculosis (TB)-burden country with high rates of Mycobacterium tuberculosis multidrug resistance (MDR) and extensive drug resistance (XDR), especially in HIV-coinfected patients. Rapid and reliable diagnosis for detection of resistance to second-line drugs is vital for adequate patient management. We evaluated the performance of the GenoType MTBDRsl (Hain Lifescience GmbH, Nehren, Germany) assay on smear-positive sputum specimens obtained from 90 HIV-infected MDR TB patients from Russia. Test interpretability was over 98%. Specificity was over 86% for all drugs, while sensitivity varied, being the highest (71.4%) for capreomycin and lowest (9.4%) for kanamycin, probably due to the presence of mutations in the eis gene. The sensitivity of detection of XDR TB was 13.6%, increasing to 42.9% if kanamycin (not commonly used in Western Europe) was excluded. The assay is a highly specific screening tool for XDR detection in direct specimens from HIV-coinfected TB patients but cannot be used to rule out XDR TB. PMID:23152552

  8. Diagnostic Accuracy of the GenoType MTBDRsl Assay for Rapid Diagnosis of Extensively Drug-Resistant Tuberculosis in HIV-Coinfected Patients

    PubMed Central

    Kontsevaya, Irina; Ignatyeva, Olga; Nikolayevskyy, Vladyslav; Balabanova, Yanina; Kovalyov, Alexander; Kritsky, Andrey; Matskevich, Olesya

    2013-01-01

    The Russian Federation is a high-tuberculosis (TB)-burden country with high rates of Mycobacterium tuberculosis multidrug resistance (MDR) and extensive drug resistance (XDR), especially in HIV-coinfected patients. Rapid and reliable diagnosis for detection of resistance to second-line drugs is vital for adequate patient management. We evaluated the performance of the GenoType MTBDRsl (Hain Lifescience GmbH, Nehren, Germany) assay on smear-positive sputum specimens obtained from 90 HIV-infected MDR TB patients from Russia. Test interpretability was over 98%. Specificity was over 86% for all drugs, while sensitivity varied, being the highest (71.4%) for capreomycin and lowest (9.4%) for kanamycin, probably due to the presence of mutations in the eis gene. The sensitivity of detection of XDR TB was 13.6%, increasing to 42.9% if kanamycin (not commonly used in Western Europe) was excluded. The assay is a highly specific screening tool for XDR detection in direct specimens from HIV-coinfected TB patients but cannot be used to rule out XDR TB. PMID:23152552

  9. Factors associated with satisfaction with community-based non-medicalized counseling and testing using HIV rapid tests among MSM in France.

    PubMed

    Préau, Marie; Lorente, Nicolas; Sagaon-Teyssier, Luis; Champenois, Karen; Gall, Jean Marie Le; Mabire, Xavier; Spire, Bruno; Mora, Marion; Yazdanpanah, Yazdan; Suzan, Marie

    2016-10-01

    The aims of the study were to determine the level of satisfaction of men who have sex with men (MSM) participating in two community-based non-medicalized counseling and testing programs (ANRS-DRAG and ANRS-COM'TEST) offering HIV rapid tests (hereafter CBOffer), and to identify factors associated with satisfaction. Between 2009 and 2011, 436 participants voluntarily benefited from a CBOffer in the two programs. They completed self-administered questionnaires before and after testing. Psychosocial scores were constructed using principal component analyses to reflect the following dimensions: post-test satisfaction, avoidance of at-risk situations as a HIV risk-reduction strategy, and attitudes towards condom use. Logarithmic regression of the post-test satisfaction score was performed on these scores and on other selected explanatory variables, including the variable "self-identification as homosexual or bisexual". Post-test satisfaction ranged between 90-99 and below 90 for 50% and 25% of the participants, respectively. Post-test satisfaction with the CBOffer was independently associated with self-defined sexuality, meeting place for sexual partners, participants' attitudes about being HIV-positive, and condom use. The very high level of satisfaction was associated with both personal and socio-behavioral factors. Vulnerable MSM could be targeted better and, accordingly, could use this offer more frequently as a combined prevention tool. PMID:27088324

  10. Rapid, Simple and Sensitive Detection of Q Fever by Loop-Mediated Isothermal Amplification of the htpAB Gene

    PubMed Central

    Pan, Lei; Zhang, Lijuan

    2013-01-01

    Background Q fever is the most widespread zoonosis, and domestic animals are the most common sources of transmission. It is not only difficult to distinguish from other febrile diseases because of the lack of specific clinical manifestations in humans, but it is also difficult to identify the disease in C. burnetii-carrying animals because of the lack of identifiable features. Conventional serodiagnosis requires sera from the acute and convalescent stages of infection, which are unavailable at early diagnosis. Nested PCR and real-time PCR require equipment. In this study, we developed a Loop-Mediated Isothermal Amplification (LAMP) assay to identify C. burnetii rapidly and sensitively. Methods A universal LAMP primer set was designed to detect the repeated sequence IS1111a of the htpAB gene of C. burnetii using PrimerExplorer V4 software. The sensitivity of the LAMP assay was evaluated using known quantities of recombined reference plasmids containing the targeted genes. The specificity of the developed LAMP assay was determined using 26 members of order Rickettsiae and 18 other common pathogens. The utility of the LAMP assay was further compared with real time PCR by the examination 24 blood samples including 6 confirmed and 18 probable Q fever cases, which diagnosed by IFA serological assessment and real time PCR. In addition, 126 animal samples from 4 provinces including 97 goats, 7 cattle, 18 horses, 3 marmots and 1 deer were compared by these two methods. Results The limits of detection of the LAMP assay for the htpAB gene were 1 copy per reaction. The specificity of the LAMP assay was 100%, and no cross-reaction was observed among the bacteria used in the study. The positive rate of unknown febrile patients was 33.3%(95%CI 30.2%–36.4%) for the LAMP assay and 8.3%(95%CI 7.4%–9.2%) for the real time PCR(P<0.05). Similarly, the total positive rate of animals was 7.9%(95%CI 7.1%–8.7%) for the LAMP assay and 0.8%(95%CI 0.7%–0.9%)for the real time PCR(P<0

  11. Simple, Rapid Mycobacterium ulcerans Disease Diagnosis from Clinical Samples by Fluorescence of Mycolactone on Thin Layer Chromatography

    PubMed Central

    Wadagni, Anita; Frimpong, Michael; Phanzu, Delphin Mavinga; Ablordey, Anthony; Kacou, Emmanuel; Gbedevi, Mirabelle; Marion, Estelle; Xing, Yalan; Babu, Vaddela Sudheer; Phillips, Richard Odame; Wansbrough-Jones, Mark; Kishi, Yoshito; Asiedu, Kingsley

    2015-01-01

    Introduction Mycobacterium ulcerans infection, known as Buruli ulcer, is a disease of the skin and subcutaneous tissues which is an important but neglected tropical disease with its major impact in rural parts of West and Central Africa where facilities for diagnosis and management are poorly developed. We evaluated fluorescent thin layer chromatography (f-TLC) for detection of mycolactone in the laboratory using samples from patients with Buruli ulcer and patients with similar lesions that gave a negative result on PCR for the IS2404 repeat sequence of M. ulcerans Methodology/Principal findings Mycolactone and DNA extracts from fine needle aspiration (FNA), swabs and biopsy specimen were used to determine the sensitivity and specificity of f-TLC when compared with PCR for the IS2404. For 71 IS2404 PCR positive and 28 PCR negative samples the sensitivity was 73.2% and specificity of 85.7% for f-TLC. The sensitivity was similar for swabs (73%), FNAs (75%) and biopsies (70%). Conclusions We have shown that mycolactone can be detected from M. ulcerans infected skin tissue by f-TLC technique. The technique is simple, easy to perform and read with minimal costs. In this study it was undertaken by a member of the group from each endemic country. It is a potentially implementable tool at the district level after evaluation in larger field studies. PMID:26583925

  12. Development of simple algorithm for direct and rapid determination of cotton maturity from FT-IR spectroscopy

    NASA Astrophysics Data System (ADS)

    Liu, Yongliang; Thibodeaux, Devron; Gamble, Gary R.

    2011-06-01

    Fourier transform infrared (FT-IR) spectra of seed and lint cottons were collected to explore the potential for the discrimination of immature cottons from mature ones and also for the determination of actual cotton maturity. Spectral features of immature and mature cottons revealed large differences in the 1200-900 cm-1 region, and such spectral distinctions formed the basis on which to develop simple three-band ratio algorithm for classification analysis. Next, an additional formula was created to assess the degree of cotton fiber maturity by converting the three-band ratios into an appropriate FT-IR maturity (MIR) index. Furthermore, the MIR index was compared with parameters derived from traditional image analysis (IA) and advanced fiber information system (AFIS) measurements. Results indicated strong correlations (R2 > 0.89) between MIR and MAFIS and between MIR and MIA among either International Cotton Calibration (ICC) standards or selected cotton maturity references. On the other hand, low correlations between the pairs were observed among regular cotton fibers, which likely resulted from the heterogeneous distribution of structural, physical, and chemical characteristics in cotton fibers and subsequent different sampling specimens for individual and independent measurement.

  13. A simple and rapid protocol to non-enzymatically dissociate fresh human tissues for the analysis of infiltrating lymphocytes.

    PubMed

    Garaud, Soizic; Gu-Trantien, Chunyan; Lodewyckx, Jean-Nicolas; Boisson, Anaïs; De Silva, Pushpamali; Buisseret, Laurence; Migliori, Edoardo; Libin, Myriam; Naveaux, Céline; Duvillier, Hugues; Willard-Gallo, Karen

    2014-01-01

    The ability of malignant cells to evade the immune system, characterized by tumor escape from both innate and adaptive immune responses, is now accepted as an important hallmark of cancer. Our research on breast cancer focuses on the active role that tumor infiltrating lymphocytes play in tumor progression and patient outcome. Toward this goal, we developed a methodology for the rapid isolation of intact lymphoid cells from normal and abnormal tissues in an effort to evaluate them proximate to their native state. Homogenates prepared using a mechanical dissociator show both increased viability and cell recovery while preserving surface receptor expression compared to enzyme-digested tissues. Furthermore, enzymatic digestion of the remaining insoluble material did not recover additional CD45(+) cells indicating that quantitative and qualitative measurements in the primary homogenate likely genuinely reflect infiltrating subpopulations in the tissue fragment. The lymphoid cells in these homogenates can be easily characterized using immunological (phenotype, proliferation, etc.) or molecular (DNA, RNA and/or protein) approaches. CD45(+) cells can also be used for subpopulation purification, in vitro expansion or cryopreservation. An additional benefit of this approach is that the primary tissue supernatant from the homogenates can be used to characterize and compare cytokines, chemokines, immunoglobulins and antigens present in normal and malignant tissues. This protocol functions extremely well for human breast tissues and should be applicable to a wide variety of normal and abnormal tissues. PMID:25548995

  14. A simple-rapid method to separate uranium, thorium, and protactinium for U-series age-dating of materials.

    PubMed

    Knight, Andrew W; Eitrheim, Eric S; Nelson, Andrew W; Nelson, Steven; Schultz, Michael K

    2014-08-01

    Uranium-series dating techniques require the isolation of radionuclides in high yields and in fractions free of impurities. Within this context, we describe a novel-rapid method for the separation and purification of U, Th, and Pa. The method takes advantage of differences in the chemistry of U, Th, and Pa, utilizing a commercially-available extraction chromatographic resin (TEVA) and standard reagents. The elution behavior of U, Th, and Pa were optimized using liquid scintillation counting techniques and fractional purity was evaluated by alpha-spectrometry. The overall method was further assessed by isotope dilution alpha-spectrometry for the preliminary age determination of an ancient carbonate sample obtained from the Lake Bonneville site in western Utah (United States). Preliminary evaluations of the method produced elemental purity of greater than 99.99% and radiochemical recoveries exceeding 90% for U and Th and 85% for Pa. Excellent purity and yields (76% for U, 96% for Th and 55% for Pa) were also obtained for the analysis of the carbonate samples and the preliminary Pa and Th ages of about 39,000 years before present are consistent with (14)C-derived age of the material. PMID:24681438

  15. A rapid and simple 8-quinolinol-based fluorescent stain of phosphoproteins in polyacrylamide gel after electrophoresis.

    PubMed

    Wang, Xu; Hwang, Sun-Young; Cong, Wei-Tao; Jin, Li-Tai; Choi, Jung-Kap

    2015-10-01

    In order to obtain an easy and rapid protocol to visualize phosphoproteins in SDS-PAGE, a fluorescent detection method named 8-Quinolinol (8-Q) stain is described. 8-Q can form ternary complexes in the gel matrix contributed by the affinity of aluminum ion (Al(3+) ) to the phosphate groups on the proteins and the metal chelating property of 8-Quinolinol, exhibiting strong fluorescence in ultraviolet light. It can visualize as little as 4∼8 ng of α-casein and β-casein, 16∼32 ng of ovalbumin and κ-casein which is more sensitive than Stains-All but less sensitive than Pro-Q Diamond. The protocol of 8-Q requires only 70 min in 0.75 mm mini-size or 1.0 mm large-size gels with five changes of solutions without destaining step; Pro-Q takes at least 250 min with 11 changes of solutions. In addition, the new method was confirmed by the study of dephosphorylation and LC-MS/MS, respectively. The approach to visualize phosphoprotein utilizing 8-Q could be an alternative to simplify the analytical operations for phosphoproteomics research. PMID:26177935

  16. Evaluation of a rapid lateral flow immunoassay for the detection of cryptococcal antigen for the early diagnosis of cryptococcosis in HIV patients in Colombia.

    PubMed

    Escandón, Patricia; Lizarazo, Jairo; Agudelo, Clara Ines; Chiller, Tom; Castañeda, Elizabeth

    2013-10-01

    A previous study carried out in a tertiary care hospital in Colombia demonstrated the usefulness of the Cryptococcus capsular antigen detection by latex (CrAg Latex) in the early diagnosis of cryptococcosis in HIV-infected patients with low CD4 + levels. The aim of this study was to establish the performance of a new rapid lateral flow assay (CrAg LFA) in preserved sera of those HIV-infected patients collected between 2001 and 2006. A total of 421 sera from 297 patients with a confirmed diagnosis of HIV were tested with CrAg LFA and results compared with those obtained with CrAg Latex. All patients provided informed consent for specimen collection. A concordance of 100% was found between positive results obtained by both methods. However, 13 sera that were negative by CrAg Latex, were positive by CrAg LFA (3.1%). In these positive patients, median of CD4 + levels was 67 cells/μl (8-608 cells/μl), while median of viral load was 118,965 copies/ml (50-500,000 copies/ml). Patients who were negative for cryptococcosis had a median of 177 cells/μl in CD4 + levels (4-2516 cells/μl) and a median of 62,318 copies/ml in viral loads (25-50,000 copies/ml). A significant statistical difference was found when comparing CD4 + levels and viral load in patients positive for cryptococcosis and those that were proven to be negative (P < 0.0001). The use of Point-of-Care Tests (POCT) like CrAg LFA play an important role in the diagnosis of infectious diseases, especially in resource limited settings, where it will be a useful means to diagnose cryptococcosis early in HIV patients. PMID:23611420

  17. A simple and rapid detection assay for peptides based on the specific recognition of aptamer and signal amplification of hybridization chain reaction.

    PubMed

    Ma, Chao; Liu, Haiyun; Tian, Tian; Song, Xianrang; Yu, Jinghua; Yan, Mei

    2016-09-15

    A simple and rapid assay for the detection of peptides is designed based on the specific recognition of aptamer, the quenching effect of graphene oxide (GO) and the efficient signal amplification of hybrid chain reaction (HCR). In this assay, the hairpin structure of aptamer is opened after binding with targets, and the initiation sequence could be exposed to hairpin probe 1 (H1) to open its hairpin structure. Then the opened H1 will open the hairpin structure of hairpin probe 2 (H2), and in turn, the opened initiation sequence of H2 continues to open H1. As a result, the specific recognition of target and fluorescent signals are accumulated through the process in short 1h. Attentively, the aptamer can not only identify target peptides, but also initiate the HCR between H1 and H2. More importantly, the HCR is initiated only after the target recognition of aptamer. After HCR, the excess hairpin probes will be anchored on the GO surface, and the background is greatly reduced due to the quenching effect of GO. By using Mucin-1(MUC1) as a model peptide, the assay has a wide linear range as two orders of magnitude and the detection range is from 0.01 to 5nM with low detection limit of 3.33pM. Therefore, the simple and rapid detection of the target can be realized, and the novel assay has great potential in detecting various peptides and even cancer cells. PMID:27093485

  18. Rapid and simple profiling of lipoproteins by polyacrylamide-gel disc electrophoresis to determine the heterogeneity of low-density lipoproteins (LDLs) including small, dense LDL.

    PubMed

    Nakano, Takanari; Inoue, Ikuo; Seo, Makoto; Takahashi, Seiichiro; Awata, Takuya; Komoda, Tsugikazu; Katayama, Shigehiro

    2009-01-01

    This study aimed to explore the potential of polyacrylamide-gel disc electrophoresis (PAGE) for lipoprotein profiling in clinical practice. Blood samples were collected from 146 patients with type 2 diabetes mellitus and lipid parameters were assayed by PAGE, including small, dense low-density lipoprotein (LDL) (n = 41), and triglyceride-rich lipoprotein remnant cholesterol (n = 37). We also used a commercial kit to measure small, dense LDL (n = 41). By PAGE, we obtained the percentage of the area under the curve (AUC %) of each peaks and calculated respective AUC% x total cholesterol (AUC%xTC) values. The calculated values of LDL-AUC%xTC, small LDL-AUC%xTC, and HDL-AUC%xTC values were correlated well with values from homogeneous assay for LDL-cholesterol, small, dense LDL-cholesterol, and HDL-cholesterol assays (r = 0.94, 0.81, and 0.89, respectively). PAGE combined with measurement of total cholesterol and triglycerides provides a rapid evaluation of anti- or pro-atherogenic lipoproteins and a simple profiling system for both the "quantity" and "quality" of lipoproteins, allowing a better assessment of the risk of coronary artery diseases. This article discusses several methods for simple and rapid lipid profiling and outlines some recent patents relevant to the methods. PMID:19149704

  19. Rapid detection of aflatoxigenic Aspergillus sp. in herbal specimens by a simple, bendable, paper-based lab-on-a-chip.

    PubMed

    Chaumpluk, Piyasak; Plubcharoensook, Pattra; Prasongsuk, Sehanat

    2016-06-01

    Postharvest herbal product contamination with mycotoxins and mycotoxin-producing fungi represents a potentially carcinogenic hazard. Aspergillus flavus is a major cause of this issue. Available mold detection methods are PCR-based and rely heavily on laboratories; thus, they are unsuitable for on-site monitoring. In this study, a bendable, paper-based lab-on-a-chip platform was developed to rapidly detect toxigenic Aspergillus spp. DNA. The 3.0-4.0 cm(2) chip is fabricated using Whatman™ filter paper, fishing line and a simple plastic lamination process and has nucleic acid amplification and signal detection components. The Aspergillus assay specifically amplifies the aflatoxin biosynthesis gene, aflR, using loop-mediated isothermal amplification (LAMP); hybridization between target DNA and probes on blue silvernanoplates (AgNPls) yields colorimetric results. Positive results are indicated by the detection pad appearing blue due to dispersed blue AgNPls; negative results are indicated by the detection pad appearing colorless or pale yellow due to probe/target DNA hybridization and AgNPls aggregation. Assay completion requires less than 40 min, has a limit of detection (LOD) of 100 aflR copies, and has high specificity (94.47%)and sensitivity (100%). Contamination was identified in 14 of 32 herbal samples tested (43.75%). This work demonstrates the fabrication of a simple, low-cost, paper-based lab-on-a-chip platform suitable for rapid-detection applications. PMID:27168276

  20. Rapid and simple identification of Beijing genotype strain of Mycobacterium tuberculosis using a loop-mediated isothermal amplification assay.

    PubMed

    Nagai, Yuhki; Iwade, Yoshito; Nakano, Manabu; Akachi, Shigehiro; Kobayashi, Takashi; Nishinaka, Takamichi

    2016-07-01

    Beijing genotype strains of Mycobacterium tuberculosis are geographically widespread and pose a notorious public health problem, these strains causing outbreaks of multidrug-resistant tuberculosis (TB); some studies have reported an association with drug resistance. Because the prevalence of Beijing strain has a substantial impact on TB control programs, the availability of a rapid and reliable method for detecting these strains is important for epidemiological monitoring of their circulation. The main methods currently used to identify Beijing genotype strains are IS6110 DNA fingerprinting, spoligotyping and PCR to detect specific deletions such as region of difference (RD)207. More recently, multiplex PCR assay using a Beijing-specific single nucleotide polymorphism (SNP) has been developed for detecting Beijing lineage strains. However, these methods are time-consuming and technically demanding. In the present study, a loop-mediated isothermal amplification (LAMP) assay that allows specific identification of Beijing genotype strain was developed. This Beijing genotype strain-identifying LAMP assay was performed 214 clinical isolates and the results compared with those of conventional PCR that targeted RD207 and Rv0679c-targreting multiplex PCR for Beijing lineage identification. LAMP assay showed 100% sensitivity and specificity compared with RD207-PCR. Furthermore, the sensitivity and specificity were 99.3% and 100%, respectively, compared with Rv0679c-multiplex PCR. This LAMP assay could be used routinely in local laboratories to monitor the prevalence of the Beijing genotype strain and thereby used to help control the spread of these potentially highly virulent and drug resistant strains. PMID:27213686

  1. Nonenzymatic amperometric response of glucose on a nanoporous gold film electrode fabricated by a rapid and simple electrochemical method.

    PubMed

    Xia, Yue; Huang, Wei; Zheng, Jufang; Niu, Zhenjiang; Li, Zelin

    2011-04-15

    An enzyme-free amperometric method was established for glucose detection using a nanoporous gold film (NPGF) electrode prepared by a rapid one-step anodic potential step method within 5 min. The prepared NPGF had an extremely high roughness and was characterized by scanning electron microscopy (SEM) and cyclic voltammetry. Electrochemical responses of the as-prepared NPGF to glucose in 0.1M phosphate buffer solution (PBS, pH 7.4) with or without Cl(-) were discussed. In amperometric studies carried out at -0.15 V in the absence of Cl(-), the NPGF electrode exhibited a high sensitivity of 232 μA mM(-1)cm(-2) and gave a linear range from 1mM up to 14 mM with a detection limit of 53.2 μM (with a signal-to-noise ratio of 3). In addition, the oxidation of ascorbic acid (AA) and uric acid (UA) can be completely eliminated at such a low applied potential. On the other hand, the quantification of glucose in 0.1M PBS (pH 7.4) containing 0.1M NaCl offered an extended linear range from 10 μM to 11 mM with a sensitivity of 66.0 μA mM(-1)cm(-2) and a low detection limit of 8.7 μM (signal-to-noise ratio of 3) at a detection potential of 0.2V. PMID:21354778

  2. Rapid and sensitive detection of Citrus Bacterial Canker by loop-mediated isothermal amplification combined with simple visual evaluation methods

    PubMed Central

    2010-01-01

    bacteria. The assay was capable of detecting CBC-causing strains from several geographical origins and pathotypes. Conclusions The CBC-LAMP technique is a simple, fast, sensitive and specific method for the diagnosis of Citrus Bacterial Canker. This method can be useful in the phytosanitary programs of the citrus industry worldwide. PMID:20565886

  3. Rapid and simple extraction of lipids from blood plasma and urine for liquid chromatography-tandem mass spectrometry.

    PubMed

    Bang, Dae Young; Byeon, Seul Kee; Moon, Myeong Hee

    2014-02-28

    A simple and fast lipid extraction method from human blood plasma and urine is introduced in this study. The effective lipid extraction from biological systems with a minimization of the matrix effect is important for the successful qualitative and quantitative analysis of lipids in liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). The method described here is based on the modification of the quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction method, which was originally developed for pesticide residue analysis in food, for the purpose of isolating lipids from biological fluids. Applicability of QuEChERS method for lipids was evaluated by varying organic solvents for the extraction/partitioning of lipids in MgSO4/CH3COONa for the removal of water and by varying sorbents (primary secondary amines, graphitized carbon black, silica, strong anion exchange resins and C18 particles) for the dispersive solid-phase extraction (dSPE) step. This study shows that 2:1 (v/v) CHCl3/CH3OH is effective in the extraction/partitioning step and that 50mg of C18 particles (for 0.1mL plasma and 1mL of urine) are more suitable for sample cleanup for the dSPE step of the QuEChERS method. Matrix effects were calculated by comparing the recovery values of lipid standards spiked to both plasma and urine samples after extraction with those of the same standards in a neat solution using nanoflow LC-ESI-MS/MS, resulting in improved MS signals due to the decrease of the ion suppression compared to the conventional Folch method. The modified QuEChERS method was applied to lipid extracts from both human urine and plasma samples, demonstrating that it can be powerfully utilized for high-speed (<15min) preparation of lipids compared to the Folch method, with equivalent or slightly improved results in lipid identification using nLC-ESI-MS/MS. PMID:24491523

  4. Simple and rapid determination of zafirlukast in plasma by ultra-performance liquid chromatography tandem mass spectrometric method: application into pharmacokinetic study in rabbits.

    PubMed

    Iqbal, M; Ezzeldin, E; Al-Rashood, K A; Al-Khamees, K I; Khan, R M A; Raish, M; Anwer, T

    2014-08-01

    Zafirlukast is a selective leukotriene receptor antagonist used for the prophylaxis and chronic treatment of asthma. The aim of the present study was to develop a simple sensitive ultra-performance liquid chromatography tandem mass spectroscopy method for rapid determination of zafirlukast in plasma. After a simple one step protein precipitation by acetonitrile, zafirlukast and montelukast (IS) were separated on Acquity UPLC BEH(TM) C18 column (50 × 2.1 mm, i.d. 1.7 µm, Waters, USA) using a mobile phase of acetonitrile:water containing 10 mM acetic acid (80:20, v/v) at a flow rate of 0.3 mL/min. Zafirlukast and IS were eluted at 0.51 and 1.1 min, respectively with a total run time of only 1.5 min. The mass spectrometric determination was carried out using an electrospray interface operated in the negative mode with multiple reactions monitoring mode. The precursor to product ion transitions of m/z 574.11>462.07 and m/z 584.2>472.1 were used to quantify zafirlukast and IS, respectively. The method was linear in the concentration range of 0.17-600 ng/mL with coefficients of determination greater than 0.996 and lower limit of quantitation of 0.17 ng/mL. Intra-day and inter-day accuracies were 88.3-113.9% and the precisions were ≤ 12.6%. Zafirlukast was found to stable under various storage and sample processing conditions as per guidelines of bio-analytical method validation. The method developed herein is simple and rapid, and was successfully applied for the pharmacokinetic study in rabbits. PMID:24258705

  5. A simple indicator to rapidly assess the short-term impact of heat waves on mortality within the French heat warning system

    NASA Astrophysics Data System (ADS)

    Antics, Annamaria; Pascal, Mathilde; Laaidi, Karine; Wagner, Vérène; Corso, Magali; Declercq, Christophe; Beaudeau, Pascal

    2013-01-01

    We propose a simple method to provide a rapid and robust estimate of the short-term impacts of heat waves on mortality, to be used for communication within a heat warning system. The excess mortality during a heat wave is defined as the difference between the observed mortality over the period and the observed mortality over the same period during the N preceding years. This method was tested on 19 French cities between 1973 and 2007. In six cities, we compared the excess mortality to that obtained using a modelling of the temperature-mortality relationship. There was a good agreement between the excess mortalities estimated by the simple indicator and by the models. Major differences were observed during the most extreme heat waves, in 1983 and 2003, and after the implementation of the heat prevention plan in 2006. Excluding these events, the mean difference between the estimates obtained by the two methods was of 13 deaths [1:45]. A comparison of mortality with the previous years provides a simple estimate of the mortality impact of heat waves. It can be used to provide early and reliable information to stakeholders of the heat prevention plan, and to select heat waves that should be further investigated.

  6. Attitude and behavior changes among gay and bisexual men after use of rapid home HIV tests to screen sexual partners.

    PubMed

    Frasca, Timothy; Balan, Ivan; Ibitoye, Mobolaji; Valladares, Juan; Dolezal, Curtis; Carballo-Diéguez, Alex

    2014-05-01

    HIV testing can now be self-administered outside clinical settings through the purchase of home testing (HT) kits. Individuals also can use the kits to perform a test on a potential sexual partner prior to intercourse. We provided a 3-month supply of HT kits to men who reported multiple male partners and little or no condom use for anal intercourse. Participants used the test kits with partners in over 100 occasions. At the end of the study, approximately half of the participants described shifts in their attitudes and/or behaviors related to sexual risk. Reported changes included increased awareness of risk, increased discussion of STI/HIV safety measures, changes in partner choice and heightened consciousness of partner thinking. Easy access to HT kits may be a risk-reduction strategy for men with a high risk profile because their regular use could have an impact beyond the specific sexual encounter. PMID:24077975

  7. Motif-Optimized Subtype A HIV Envelope-based DNA Vaccines Rapidly Elicit Neutralizing Antibodies When Delivered Sequentially

    PubMed Central

    Pissani, Franco; Malherbe, Delphine C.; Robins, Harlan; DeFilippis, Victor R.; Park, Byung; Sellhorn, George; Stamatatos, Leonidas; Overbaugh, Julie; Haigwood, Nancy L.

    2012-01-01

    HIV-1 infection results in the development of a diverging quasispecies unique to each infected individual. Envelope (Env)-specific neutralizing antibodies (NAbs) typically develop over months to years after infection and initially are limited to the infecting virus. In some subjects, antibody responses develop that neutralize heterologous isolates (HNAbs), a phenomenon termed broadening of the NAb response. Studies of co-crystalized antibodies and proteins have facilitated the identification of some targets of broadly neutralizing monoclonal antibodies (NmAbs) capable of neutralizing many or most heterologous viruses; however, the ontogeny of these antibodies in vivo remains elusive. We hypothesize that Env protein escape variants stimulate broad NAb development in vivo and could generate such NAbs when used as immunogens. Here we test this hypothesis in rabbits using HIV Env vaccines featuring: (1) use of individual quasispecies env variants derived from an HIV-1 subtype A-infected subject exhibiting high levels of NAbs within the first year of infection that increased and broadened with time; (2) motif optimization of envs to enhance in vivo expression of DNA formulated as vaccines; and (3) a combined DNA plus protein boosting regimen. Vaccines consisted of multiple env variants delivered sequentially and a simpler regimen that utilized only the least and most divergent clones. The simpler regimen was as effective as the more complex approach in generating modest HNAbs and was more efficient when modified, motif-optimized DNA was used in combination with trimeric gp140 protein. This is a rationally designed strategy that facilitates future vaccine design by addressing the difficult problem of generating HNAbs to HIV by empirically testing the immunogenicity of naturally occurring quasispecies env variants. PMID:22749601

  8. Simple and rapid method for the determination of ethylenebisdithiocarbamate fungicides in fruits and vegetables using liquid chromatography with tandem mass spectrometry.

    PubMed

    Hayama, Tadashi; Takada, Makoto

    2008-11-01

    A simple and rapid method for determining ethylenebisdithiocarbamates (EBDCs; mancozeb, maneb, and zineb) in fruits and vegetables is described. EBDCs are transformed into dimethylethylenebisdithiocarbamate (EBDC-dimethyl) by methylation after their decomposition with ethylenediaminetetraacetic acid (EDTA). These processes were performed simultaneously in this method. Dimethyl sulfate was used as the methylation reagent, and acetonitrile extracts obtained from partitioning with anhydrous magnesium sulfate and sodium chloride were subjected to dispersive solid-phase extraction with the primary secondary amine sorbent. Liquid chromatography with tandem mass spectrometry in the positive heated-electrospray ionization mode was used for the determination of EBDC-dimethyl produced from EBDCs. The method was validated at levels of 10, 50, and 100 ng g(-1) maneb as a representative EBDC. The recoveries of the present method were between 71 and 101%. The limits of detection and quantification were 0.24 and 0.8 ng g(-1) maneb, respectively. PMID:18759101

  9. Expert System Shells for Rapid Clinical Decision Support Module Development: An ESTA Demonstration of a Simple Rule-Based System for the Diagnosis of Vaginal Discharge

    PubMed Central

    2012-01-01

    Objectives This study demonstrates the feasibility of using expert system shells for rapid clinical decision support module development. Methods A readily available expert system shell was used to build a simple rule-based system for the crude diagnosis of vaginal discharge. Pictures and 'canned text explanations' are extensively used throughout the program to enhance its intuitiveness and educational dimension. All the steps involved in developing the system are documented. Results The system runs under Microsoft Windows and is available as a free download at http://healthcybermap.org/vagdisch.zip (the distribution archive includes both the program's executable and the commented knowledge base source as a text document). The limitations of the demonstration system, such as the lack of provisions for assessing uncertainty or various degrees of severity of a sign or symptom, are discussed in detail. Ways of improving the system, such as porting it to the Web and packaging it as an app for smartphones and tablets, are also presented. Conclusions An easy-to-use expert system shell enables clinicians to rapidly become their own 'knowledge engineers' and develop concise evidence-based decision support modules of simple to moderate complexity, targeting clinical practitioners, medical and nursing students, as well as patients, their lay carers and the general public (where appropriate). In the spirit of the social Web, it is hoped that an online repository can be created to peer review, share and re-use knowledge base modules covering various clinical problems and algorithms, as a service to the clinical community. PMID:23346475

  10. Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor.

    PubMed

    Song, Chunmei; Li, Jianwu; Liu, Jinxin; Liu, Qing

    2016-08-15

    A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 10(6)cells/mL while the LOD for semi-quantitative detection could go down to 10(5)cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing. PMID:27260433

  11. A fast transfer-free synthesis of high-quality monolayer graphene on insulating substrates by a simple rapid thermal treatment.

    PubMed

    Wu, Zefei; Guo, Yanqing; Guo, Yuzheng; Huang, Rui; Xu, Shuigang; Song, Jie; Lu, Huanhuan; Lin, Zhenxu; Han, Yu; Li, Hongliang; Han, Tianyi; Lin, Jiangxiazi; Wu, Yingying; Long, Gen; Cai, Yuan; Cheng, Chun; Su, Dangsheng; Robertson, John; Wang, Ning

    2016-02-01

    The transfer-free synthesis of high-quality, large-area graphene on a given dielectric substrate, which is highly desirable for device applications, remains a significant challenge. In this paper, we report on a simple rapid thermal treatment (RTT) method for the fast and direct growth of high-quality, large-scale monolayer graphene on a SiO2/Si substrate from solid carbon sources. The stack structure of a solid carbon layer/copper film/SiO2 is adopted in the RTT process. The inserted copper film does not only act as an active catalyst for the carbon precursor but also serves as a "filter" that prevents premature carbon dissolution, and thus, contributes to graphene growth on SiO2/Si. The produced graphene exhibits a high carrier mobility of up to 3000 cm(2) V(-1) s(-1) at room temperature and standard half-integer quantum oscillations. Our work provides a promising simple transfer-free approach using solid carbon sources to obtain high-quality graphene for practical applications. PMID:26499039

  12. A fast transfer-free synthesis of high-quality monolayer graphene on insulating substrates by a simple rapid thermal treatment

    NASA Astrophysics Data System (ADS)

    Wu, Zefei; Guo, Yanqing; Guo, Yuzheng; Huang, Rui; Xu, Shuigang; Song, Jie; Lu, Huanhuan; Lin, Zhenxu; Han, Yu; Li, Hongliang; Han, Tianyi; Lin, Jiangxiazi; Wu, Yingying; Long, Gen; Cai, Yuan; Cheng, Chun; Su, Dangsheng; Robertson, John; Wang, Ning

    2016-01-01

    The transfer-free synthesis of high-quality, large-area graphene on a given dielectric substrate, which is highly desirable for device applications, remains a significant challenge. In this paper, we report on a simple rapid thermal treatment (RTT) method for the fast and direct growth of high-quality, large-scale monolayer graphene on a SiO2/Si substrate from solid carbon sources. The stack structure of a solid carbon layer/copper film/SiO2 is adopted in the RTT process. The inserted copper film does not only act as an active catalyst for the carbon precursor but also serves as a ``filter'' that prevents premature carbon dissolution, and thus, contributes to graphene growth on SiO2/Si. The produced graphene exhibits a high carrier mobility of up to 3000 cm2 V-1 s-1 at room temperature and standard half-integer quantum oscillations. Our work provides a promising simple transfer-free approach using solid carbon sources to obtain high-quality graphene for practical applications.

  13. Successful combination of computationally inexpensive GIAO 13C NMR calculations and artificial neural network pattern recognition: a new strategy for simple and rapid detection of structural misassignments.

    PubMed

    Sarotti, Ariel M

    2013-08-01

    GIAO NMR chemical shift calculations coupled with trained artificial neural networks (ANNs) have been shown to provide a powerful strategy for simple, rapid and reliable identification of structural misassignments of organic compounds using only one set of both computational and experimental data. The geometry optimization, usually the most time-consuming step in the overall procedure, was carried out using computationally inexpensive methods (MM+, AM1 or HF/3-21G) and the NMR shielding constants at the affordable mPW1PW91/6-31G(d) level of theory. As low quality NMR prediction is typically obtained with such protocols, the decision making was foreseen as a problem of pattern recognition. Thus, given a set of statistical parameters computed after correlation between experimental and calculated chemical shifts the classification was done using the knowledge derived from trained ANNs. The training process was carried out with a set of 200 molecules chosen to provide a wide array of chemical functionalities and molecular complexity, and the results were validated with a set of 26 natural products that had been incorrectly assigned along with their 26 revised structures. The high prediction effectiveness observed makes this method a suitable test for rapid identification of structural misassignments, preventing not only the publication of wrong structures but also avoiding the consequences of such a mistake. PMID:23779148

  14. A Simple and Rapid LC-MS/MS Method for the Determination of BMCL26 a Novel Anti-Parasitic Agent in Rat Plasma

    PubMed Central

    Voggu, Ramakrishna R; Zhou, Xiang; Su, Bin; Guo, Baochuan

    2015-01-01

    BMCL26 is a potential drug derived from nimesulide, which has exhibited the substantial anti-parasitic activity in various cell lines. To conduct various pharmacological and toxicological properties of this drug, we developed and validated a rapid LC-MS/MS method for its quantification in accordance with the FDA guidelines. Protein precipitation with 0.1% formic acid in acetonitrile was used to extract the analytes along with the internal standard (JCC76) from rat plasma. It was found that the calibration curve of the method had an excellent linearity (r2 ≥ 0.9993) for the analyte concentration ranging from 0.5 to 100 ng/mL with acceptable inter- and intra-assay, precision, accuracy and stability. The matrix effect and extraction recovery were in the range of 101.30–110.10% and 90.16– 105.00%, respectively. This LC-MS/MS method is simple and rapid and can be used in the future pharmaceutical studies of BMCL26. PMID:26823991

  15. Establishment and application of a loop-mediated isothermal amplification method for simple, specific, sensitive and rapid detection of Toxoplasma gondii.

    PubMed

    Cao, Lili; Cheng, Ronghua; Yao, Lin; Yuan, Shuxian; Yao, Xinhua

    2014-01-01

    The Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high simply, specificity, sensitivity and rapidity. In this study, A LAMP assay with 6 primers targeting a highly conserved region of the GRA1 gene was developed to diagnose Toxoplasma gondii. The reaction time of the LAMP assay was shortened to 30 min after optimizing the reaction system. The LAMP assay was found to be highly specific and stable. The detection limit of the LAMP assay was 10 copies, the same as that of the conventional PCR. We used the LAMP assay to develop a real-time fluorogenic protocol to quantitate T. gondii DNA and generated a log-linear regression plot by plotting the time-to-threshold values against genomic equivalent copies. Furthermore, the LAMP assay was applied to detect T. gondii DNA in 423 blood samples and 380 lymph node samples from 10 pig farms, and positive results were obtained for 7.8% and 8.2% of samples, respectively. The results showed that the LAMP method is slightly more sensitive than conventional PCR (6.1% and 7.6%). Positive samples obtained from 6 pig farms. The LAMP assay established in this study resulted in simple, specific, sensitive and rapid detection of T. gondii DNA and is expected to play an important role in clinical detection of T. gondii. PMID:23965849

  16. Rapid Implementation of an Integrated Large-Scale HIV Counseling and Testing, Malaria, and Diarrhea Prevention Campaign in Rural Kenya

    PubMed Central

    Lugada, Eric; Millar, Debra; Haskew, John; Grabowsky, Mark; Garg, Navneet; Vestergaard, Mikkel; Kahn, James; Muraguri, Nicholas; Mermin, Jonathan

    2010-01-01

    Background Integrated disease prevention in low resource settings can increase coverage, equity and efficiency in controlling high burden infectious diseases. A public-private partnership with the Ministry of Health, CDC, Vestergaard Frandsen and CHF International implemented a one-week integrated multi-disease prevention campaign. Method Residents of Lurambi, Western Kenya were eligible for participation. The aim was to offer services to at least 80% of those aged 15–49. 31 temporary sites in strategically dispersed locations offered: HIV counseling and testing, 60 male condoms, an insecticide-treated bednet, a household water filter for women or an individual filter for men, and for those testing positive, a 3-month supply of cotrimoxazole and referral for follow-up care and treatment. Findings Over 7 days, 47,311 people attended the campaign with a 96% uptake of the multi-disease preventive package. Of these, 99.7% were tested for HIV (87% in the target 15–49 age group); 80% had previously never tested. 4% of those tested were positive, 61% were women (5% of women and 3% of men), 6% had median CD4 counts of 541 cell/µL (IQR; 356, 754). 386 certified counselors attended to an average 17 participants per day, consistent with recommended national figures for mass campaigns. Among women, HIV infection varied by age, and was more likely with an ended marriage (e.g. widowed vs. never married, OR.3.91; 95% CI. 2.87–5.34), and lack of occupation. In men, quantitatively stronger relationships were found (e.g. widowed vs. never married, OR.7.0; 95% CI. 3.5–13.9). Always using condoms with a non-steady partner was more common among HIV-infected women participants who knew their status compared to those who did not (OR.5.4 95% CI. 2.3–12.8). Conclusion Through integrated campaigns it is feasible to efficiently cover large proportions of eligible adults in rural underserved communities with multiple disease preventive services simultaneously achieving various

  17. Evaluation of the Rapid Scale-up of Collaborative TB/HIV Activities in TB Facilities in Rwanda, 2005-2009

    PubMed Central

    2011-01-01

    Background In 2005, Rwanda drafted a national TB/HIV policy and began scaling-up collaborative TB/HIV activities. Prior to the scale-up, we evaluated existing TB/HIV practices, possible barriers to policy and programmatic implementation, and patient treatment outcomes. We then used our evaluation data as a baseline for evaluating the national scale-up of collaborative TB/HIV activities from 2005 through 2009. Methods Our baseline evaluation included a cross-sectional evaluation of 23/161 TB clinics. We conducted structured interviews with patients and clinic staff and reviewed TB registers and patient records to assess HIV testing practices, provision of HIV care and treatment for people with TB that tested positive for HIV, and patients' TB treatment outcomes. Following our baseline evaluation, we used nationally representative TB/HIV surveillance data to monitor the scale-up of collaborative TB/HIV activities Results Of 207 patients interviewed, 76% were offered HIV testing, 99% accepted, and 49% reported positive test results. Of 40 staff interviewed, 68% reported offering HIV testing to >50% of patients. From 2005-2009, scaled-up TB/HIV activities resulted in increased HIV testing of patients with TB (69% to 97%) and provision of cotrimoxazole (15% to 92%) and antiretroviral therapy (13% to 49%) for patients with TB disease and HIV infection (TB/HIV). The risk of death among patients with TB/HIV relative to patients with TB not infected with HIV declined from 2005 (RR = 6.1, 95%CI 2.6, 14.0) to 2007 (RR = 1.8, 95%CI 1.68, 1.94). Conclusions Our baseline evaluation highlighted that staff and patients were receptive to HIV testing. However, expanded access to testing, care, and treatment was needed based on the proportion of patients with TB having unknown HIV status and the high rate of HIV infection and poorer TB treatment outcomes for patients with TB/HIV. Following our evaluation, scale-up of TB/HIV services resulted in almost all patients with TB knowing

  18. Photocatalytic reduction of CO2 over Ag/TiO2 nanocomposites prepared with a simple and rapid silver mirror method

    NASA Astrophysics Data System (ADS)

    Yu, Bingcheng; Zhou, Yong; Li, Peng; Tu, Wenguang; Li, Ping; Tang, Lanqin; Ye, Jinhua; Zou, Zhigang

    2016-06-01

    The photocatalytic reduction of CO2 over Ag/TiO2 composites prepared with a simple silver mirror reaction method was investigated under UV-visible irradiation in both gas-phase (CO2 + water vapor) and aqueous solution (CO2-saturated NaHCO3 solution) systems. The as-prepared Ag/TiO2 nanocomposite exhibits efficient photocatalytic activity due to the surface plasmonic resonance and electron sink effect of the Ag component, which was found to be closely related to the size and loading amount of Ag. The rapid silver method is effective at curbing the size of Ag, so photocatalytic activity can be improved. Diverse organic chemical products were detected, including mainly methane and methanol as well as a small amount of C2 and C3 species such as acetaldehyde and acetone. Possible photocatalytic mechanisms were proposed. This artificial photosynthesis process may give a prosperous route to the removal of CO2 while simultaneously converting CO2 to valuable fuels based on highly efficient photocatalysts.The photocatalytic reduction of CO2 over Ag/TiO2 composites prepared with a simple silver mirror reaction method was investigated under UV-visible irradiation in both gas-phase (CO2 + water vapor) and aqueous solution (CO2-saturated NaHCO3 solution) systems. The as-prepared Ag/TiO2 nanocomposite exhibits efficient photocatalytic activity due to the surface plasmonic resonance and electron sink effect of the Ag component, which was found to be closely related to the size and loading amount of Ag. The rapid silver method is effective at curbing the size of Ag, so photocatalytic activity can be improved. Diverse organic chemical products were detected, including mainly methane and methanol as well as a small amount of C2 and C3 species such as acetaldehyde and acetone. Possible photocatalytic mechanisms were proposed. This artificial photosynthesis process may give a prosperous route to the removal of CO2 while simultaneously converting CO2 to valuable fuels based on highly

  19. Selective colorimetric sensors based on the monitoring of an unmodified silver nanoparticles (AgNPs) reduction for a simple and rapid determination of mercury

    NASA Astrophysics Data System (ADS)

    Jarujamrus, Purim; Amatatongchai, Maliwan; Thima, Araya; Khongrangdee, Thatsanee; Mongkontong, Chakrit

    2015-05-01

    In this work, selective colorimetric sensors for simple and rapid detection of Hg(II) ions based on the monitoring of an unmodified silver nanoparticles (AgNPs) reduction were developed. The average diameter of synthesized AgNPs was 8.3 ± 1.4 nm which was characterized by transmission electron microscopy (TEM). The abrupt change in absorbance of the unmodified AgNPs was observed which progressively decreased and slightly shifted to the blue wavelength as the concentration of Hg(II) increased, indicating the oxidation of Ag(0) to Ag(I) occurred. It appears that the AgNPs were oxidized by Hg(II), resulting in disintegration of the AgNPs into smaller particles as well as mediating the reduction of Hg(II) to Hg(0) adsorbed onto the surface of AgNPs. The adsorption of Hg(0) resulted in the lack of sufficient charges on AgNPs surfaces due to the decrease in the surface coverage of negatively charged citrate molecules, which then leaded to enlargement of AgNPs. The calibration curve of this technique was demonstrated from 0.5 to 7 ppm (r2 = 0.995), the limit of detection (LOD) was 0.06 ppm (SDblank/slope of calibration curve) with the precision (RSD, n = 4) of 3.24-4.53. Interestingly, the results show a significant enhance in the Hg(II) analytical sensitivity when Cu(II) is doped onto the unmodified AgNPs, which improves the quantitative detection limit to 0.008 ppm. In addition, greater selectivity toward Hg(II) compared with the other metal ions tested was observed. Furthermore, the percentage recoveries of spiked drinking water, tap water and SRM1641d (mercury in water) were in acceptable range with a good precision (RSD) which were in agreement with the values obtained from graphite furnace atomic absorption spectrometer (GFAAS). The technique proposed in this study provides a rapid, simple, sensitive and selective detection method for Hg(II) in water samples.

  20. Simple and rapid determination of phthalates using microextraction by packed sorbent and gas chromatography with mass spectrometry quantification in cold drink and cosmetic samples.

    PubMed

    Kaur, Ramandeep; Heena; Kaur, Ripneel; Rani, Susheela; Malik, Ashok Kumar

    2016-03-01

    A simple and rapid method using microextraction by packed sorbent coupled with gas chromatography and mass spectrometry has been developed for the analysis of five phthalates, namely, diethyl phthalate, benzyl-n-butyl phthalate, dicyclohexyl phthalate, di-n-butyl phthalate, and di-n-propyl phthalate, in cold drink and cosmetic samples. The various parameters that influence the microextraction by packed sorbent performance such as extraction cycle (extract-discard), type and amount of solvent, washing solvent, and pH have been studied. The optimal conditions of microextraction using C18 as the packed sorbent were 15 extraction cycles with water as washing solvent and 3 × 10 μL of ethyl acetate as the eluting solvent. Chromatographic separation was also optimized for injection temperature, flow rate, ion source, interface temperature, column temperature gradient and mass spectrometry was evaluated using the scan and selected ion monitoring data acquisition mode. Satisfactory results were obtained in terms of linearity with R(2) >0.9992 within the established concentration range. The limit of detection was 0.003-0.015 ng/mL, and the limit of quantification was 0.009-0.049 ng/mL. The recoveries were in the range of 92.35-98.90% for cold drink, 88.23-169.20% for perfume, and 88.90-184.40% for cream. Analysis by microextraction by packed sorbent promises to be a rapid method for the determination of these phthalates in cold drink and cosmetic samples, reducing the amount of sample, solvent, time and cost. PMID:26683135

  1. Assessment of an outreach street-based HIV rapid testing programme as a strategy to promote early diagnosis: a comparison with two surveillance systems in Spain, 2008-2011.

    PubMed

    Belza, M J; Hoyos, J; Fernández-Balbuena, S; Diaz, A; Bravo, M J; de la Fuente, L

    2015-01-01

    We assess the added value of a multisite, street-based HIV rapid testing programme by comparing its results to pre-existing services and assessing its potential to reduce ongoing transmission. Between 2008 and 2011, 8,923 individuals underwent testing. We compare outcomes with those of a network of 20 sexually transmitted infections (STI)/HIV clinics (EPI-VIH) and the Spanish National HIV Surveillance System (SNHSS); evaluate whether good visibility prompts testing and assess whether it reaches under-tested populations. 89.2% of the new infections were in men who have sex with men (MSM) vs 78.0% in EPI-VIH and 56.0% in SNHSS. 83.6% of the MSM were linked to care and 20.9% had <350 CD4 HIV prevalence was substantially lower than in EPI-VIH. 56.5% of the HIV-positive MSM tested because they happened to see the programme, 18.4% were previously untested and 26.3% had their last test ≥2 years ago. The programme provided linkage to care and early diagnosis mainly to MSM but attendees presented a lower HIV prevalence than EPI-VIH. From a cost perspective it would benefit from being implemented in locations highly frequented by MSM. Conversely, its good visibility led to reduced periods of undiagnosed infection in a high proportion of MSM who were not testing with the recommended frequency. PMID:25884149

  2. Use of amplification refractory mutation system PCR assay as a simple and effective tool to detect HIV-1 drug resistance mutations.

    PubMed

    Nanfack, Aubin J; Agyingi, Lucy; Noubiap, Jean Jacques N; Ngai, Johnson N; Colizzi, Vittorio; Nyambi, Phillipe N

    2015-05-01

    Access to genotyping assays to determine successful antiretroviral treatment (ART) is limited in resource-constrained settings by high cost, suggesting the need for a cost-effective and simplified method to identify HIV-1 drug resistance (HIVDR) mutations. In this study, an amplification refractory mutation system (ARMS)-PCR assay was developed and used to investigate the most frequent HIVDR mutations affecting first-line ART in settings where WHO ART guidelines are applied. Seventy-five HIV-positive (HIV(+)) samples from Cameroon were used to assess the performance of this assay. Sequencing of HIV-1 reverse transcriptase was simultaneously performed for comparison, and discordant samples were tested with a Trugene HIV-1 genotyping kit. The ARMS-PCR assay was able to detect M184V, T215Y/F, K103N, and Y181C mutations with sensitivities of 96.8%, 85.7%, 91.3%, and 70%, respectively, and specificities of 90.6%, 95%, 100%, 96.9%, respectively, compared with data on sequencing. The results indicated the highest positive predictive value for K103N (100%) and the highest negative predictive value for M184V (97.5%). ARMS-PCR's limits of detection for mutations M184V, T215Y/F, K103N, and Y181C were <75 copies/ml, 143 copies/ml, 143 copies/ml, and 836 copies/ml, respectively. ARMS-PCR efficiently identified mutations in individuals harboring different HIV-1 clades (CRF02_AG and non-CRF02_AG). In addition, this approach was more cost-effective than other genotyping assays. The high throughput, the cost-effectiveness, and the simplicity of the ARMS-PCR assay make it a suitable tool to monitor HIVDR patterns in resource-constrained settings with broad HIV-1 genetic diversity. PMID:25788547

  3. A simple and rapid screening method for sulfonamides in honey using a flow injection system coupled to a liquid waveguide capillary cell.

    PubMed

    Catelani, Tiago Augusto; Tóth, Ildikó Vargáné; Lima, José L F C; Pezza, Leonardo; Pezza, Helena Redigolo

    2014-04-01

    A rapid and simple screening method was developed for the determination of sulfonamides in honey samples by flow injection analysis (FIA) coupled to a liquid waveguide capillary cell. The proposed method is based on the reaction between sulfonamides and p-dimethylaminocinnamaldehyde (p-DAC) in the presence of sodium dodecylsulate (SDS) in dilute acid medium (hydrochloric acid), with the reaction product being measured spectrophotometrically at λ(max) = 565 nm. Experimental design methodology was used to optimize the analytical conditions. The proposed technique was applied to the determination of sulfonamides (sulfaquinoxaline, sulfadimethoxine, and sulfathiazole) in honey samples, in a concentration range from 6.00 × 10(-3) to 1.15 × 10(-1)mg L(-1). The detection (LOD) and quantification (LOQ) limits were 1.66 × 10(-3) and 5.54 × 10(-3)mg L(-1), respectively. Positive and false positive samples were also analyzed by a confirmatory HPLC method. The proposed system enables the screening of sulfonamides in honey samples with a low number of false positive results, with fast response therefore offers a new tool for consumer protection. PMID:24607139

  4. A rapid and simple UPLC-MS-MS method for determination of glipizide in human plasma and its application to bioequivalence study.

    PubMed

    Qiu, Xiangjun; Zheng, Shuang-li; Wang, Yingfei; Wang, Rong; Ye, Lei

    2015-01-01

    In this study, a simple, rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry method is described for the determination of glipizide in human plasma samples using carbamazepine as the internal standard (IS) from bioequivalence assays. Sample preparation was accomplished through protein precipitation with methanol, and chromatographic separation was performed on an Acquity BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with gradient profile at a flow rate of 0.4 mL/min. Mass spectrometric analysis was performed using an QTrap5500 mass spectrometer coupled with an electrospray ionization source in the positive ion mode. The multiple reaction monitoring transitions of m/z 446.1 → 321.0 and m/z 237.1 → 194.2 were used to quantify for glipizide and IS. The linearity of this method was found to be within the concentration range of 10-1,500 ng/mL for glipizide in human plasma. Only 1.0 min was needed for an analytical run. The method was applied to a bioequivalence study of two drug products containing glipizide in human plasma samples. PMID:24771054

  5. Development and Validation of a Rapid and Simple UPLC-ESI-MS Method for Pharmacokinetics and Tissue Distribution of Astragaloside III in Rats.

    PubMed

    Liu, Xiao-Hua; Guo, Long; Yang, Ying-Lai; Hu, Fang; Chen, Xin-Yue; Feng, Shi-Lan

    2016-05-01

    A rapid and simple ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) method for the determination of astragaloside III was developed and used in a pharmacokinetic and tissue distribution study in rats following the oral administration 95% ethanol extraction of Zhenqi Fuzheng capsules. Although astragaloside III and astragaloside IV have the same molecular weight and very similar structures, they were successfully separated using this method. Quantification was performed using low-energy collision tandem mass spectrometry (CID-MS-MS) with the multiple reaction monitoring scan mode of the following precursor ion → product ion atm/z807.61→335.22 for astragaloside III and atm/z633.18→331.18 for the internal standard (hesperidin). Both astragaloside III and astragaloside IV in rat plasma were best fit to a two-compartment model. The tissue distribution study showed the overall trend of disposition of astragaloside III were Cthymus> Cspleen> Cstomach> Cliver> Cheart> Ckidney> Clung> Ctesticle The high levels of astragaloside III in thymus and spleen indicated an accumulation in organs involved in immune responses and showed that these organs are major target sitesin vivo The results in the article will provide valuable information for use in clinical applications of astragaloside III. PMID:26931734

  6. A simple and rapid ultra-high-performance liquid chromatography-tandem mass spectrometry method to determine plasma biotin in hemodialysis patients.

    PubMed

    Yagi, Shigeaki; Nishizawa, Manabu; Ando, Itiro; Oguma, Shiro; Sato, Emiko; Imai, Yutaka; Fujiwara, Masako

    2016-08-01

    A simple, rapid, and selective method for determination of plasma biotin was developed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). After single-step protein precipitation with methanol, biotin and stable isotope-labeled biotin as an internal standard (IS) were chromatographed on a pentafluorophenyl stationary-phase column (2.1 × 100 mm, 2.7 μm) under isocratic conditions using 10 mm ammonium formate-acetonitrile (93:7, v/v) at a flow rate of 0.6 mL/min. The total chromatographic runtime was 5 min for each injection. Detection was performed in a positive electrospray ionization mode by monitoring selected ion transitions at m/z 245.1/227.0 and 249.1/231.0 for biotin and the IS, respectively. The calibration curve was linear in the range of 0.05-2 ng/mL using 300 μL of plasma. The intra- and inter-day precisions were all <7.1%. The accuracy varied from -0.7 to 8.2%. The developed UHPLC-MS/MS method was successfully applied to determine plasma biotin concentrations in hemodialysis patients. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26715368

  7. A Simple and Rapid Method Based on Anti-aggregation of Silver Nanoparticles for Detection of Poly(diallyldimethylammonium chloride) in Tap Water.

    PubMed

    Trisaranakul, Wichaya; Chompoosor, Apiwat; Maneeprakorn, Weerakanya; Nacapricha, Duangjai; Choengchan, Nathawut; Teerasong, Saowapak

    2016-01-01

    A simple and rapid method was developed for the detection of poly(diallyldimethylammonium chloride) (PDADMAC) using citrate-capped silver nanoparticles (AgNPs). Detection was based on anti-aggregation of AgNPs in phosphate buffer caused by PDADMAC. Due to its positive charges, PDADMAC was adsorbed onto AgNPs via electrostatic interaction with citrate, which resulted in the charges at the particle surfaces to become positive and caused repulsion among particles. Furthermore, long-chain PDADMAC provided steric hindrance. These two effects promoted the dispersion of AgNPs in the phosphate buffer. A change in the state of dispersion influenced the surface plasmon resonance (SPR) of AgNPs. Therefore, in this work, the concentration of PDADMAC was determined by monitoring changes in absorbance (at 396 nm) caused by SPR of AgNPs. Under optimal conditions, the calibration was linear over the range of 1 to 100 mg L(-1) with a detection limit of 0.7 mg L(-1). Satisfactory precision was obtained (RSD = 2.8%). This method was successfully applied to the determination of PDADMAC in tap water samples. The recoveries ranged from 86.0 - 107.5%. PMID:27396659

  8. A Simple and Rapid UPLC Method for the Determination of Rosavin in Rat Plasma and Its Application to a Pharmacokinetic Study.

    PubMed

    Chen, Dahui; Sun, Hao; Shen, Jiaqi; Igor, Longo Phemba; Zheng, Xiaoyong; Hu, Shuping; Xiang, Zheng

    2016-08-01

    Rosavin is a bioactive antidepressant component isolated from Rhodiola rosea L. In this work, an ultra-performance liquid chromatography (UPLC) method was established for the determination of rosavin in rat plasma. The chromatographic separation was achieved on a HSS T3 column (100 mm × 2.1 mm, 1.8 μm) with a gradient mobile phase consisting of acetonitrile and water (0.1% formic acid). Plasma samples were processed with one-step protein precipitation. Rutin was chosen as internal standard and the detection wavelength was 249 nm. The pharmacokinetic parameters were analyzed using the drug and statistics software. The results showed that the established method has an excellent linearity in the range of 10-1,000 ng/mL (R(2) = 0.992) with a lower limit of quantification (10 ng/mL). The intra- and interday precision (relative standard deviation) were from 2.0 to 10.6% and the extraction recovery was 92.4-95.1%. The simple and rapid UPLC method was successfully applied to the pharmacokinetic and bioavailability study of rosavin in rats. PMID:27048645

  9. Rapid and simple method for determination of cephradine in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS): application to the bioequivalence study.

    PubMed

    Choi, Sang-Jun; Ryu, Ju-Hee; Lee, Heon-Woo; Lee, Myung-Jae; Seo, Ji-Hyung; Tak, Seong-Kun; Lee, Kyung-Tae

    2009-12-01

    A rapid and simple procedure was developed for the determination of cephradine in human plasma using liquid chromatography coupled with electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). After trichloroacetic acid (TCA) precipitation of proteins from plasma samples, cephradine and cefaclor (the internal standard; IS) were eluted on a CN column. The isocratic mobile phase used consisted of acetonitrile-water-formic acid (25:75:0.1, v/v/v). Cephradine and the IS were both detected in multiple reaction monitoring (MRM) mode at the transitions: m/z 350.0 --> 90.8 for cephradine and m/z 368.1 --> 106.0 for the IS, respectively. The calibration curve was linear over the concentration range from 0.05 to 50 microg/ml, and correlation coefficients were greater than 0.996. The coefficient of variation of assay precision was less than 9.36%, and its accuracy ranged from 87.92% to 111.16%. The chromatographic run time for each plasma sample was less than 3 min. The developed method was successfully applied to a bioequivalence study of cephradine in healthy male volunteers. PMID:19854118

  10. A simple and rapid method for measuring unconjugated capsular polysaccharide (PRP) of Haemophilus influenzae type b in PRP-tetanus toxoid conjugate vaccine.

    PubMed

    Guo, Y Y; Anderson, R; McIver, J; Gupta, R K; Siber, G R

    1998-03-01

    The authors developed a simple and rapid method for quantitation of free capsular polysaccharide of Haemophilus influenzae type b (polyribosyl ribitol phosphate, PRP) in PRP-tetanus toxoid conjugate vaccine based on acid precipitation of tetanus toxoid (TT). Acid hydrolysis of PRP during the assay was not detected. The conditions used in the assay did not precipitate unconjugated PRP or adipic acid dihydrazide derivatized PRP. The method was highly reliable, reproducible and sensitive. The accuracy of the assay was confirmed by spiking known amounts of unconjugated PRP to PRP-TT conjugate preparations. A PRP-TT preparation, incubated at 37 degrees C for 6 months showing most of the PRP as unconjugated (87% determined by this method), was not immunogenic in mice for the PRP component even after two injections. In contrast, the same preparation held at 4 degrees C for 20 months, showing 17% unconjugated PRP, induced IgG antibodies to PRP which were boosted after second injection. Therefore, this method is very useful to evaluate the stability of PRP-TT conjugate vaccine. The assay may be useful for characterizing other polysaccharide-protein conjugate vaccines. PMID:9637747

  11. A simple and rapid method for HLA-DQA1 genotyping by digestion of PCR-amplified DNA with allele specific restriction endonucleases.

    PubMed

    Maeda, M; Murayama, N; Ishii, H; Uryu, N; Ota, M; Tsuji, K; Inoko, H

    1989-11-01

    The second exon of the HLA-DQA1 genes was selectively amplified from genomic DNAs of 72 HLA-homozygous B cell lines by the polymerase chain reaction (PCR). Amplified DNAs were digested with HaeIII, Ddel, ScrFI, FokI and RsaI, which recognize allelic sequence variations in the polymorphic segments of the DQA1 second exon, and then subjected to electrophoresis in polyacrylamide gels. Eight different polymorphic patterns of restriction fragments were obtained, and seven were identical to patterns predicted from the known DNA sequences, correlating with each HLA-DQw type defined by serological typing. The remaining one pattern cannot be explained from the sequence data, suggesting the presence of a novel DQA1 allele at the nucleotide level. This PCR-RFLP method provides a simple and rapid technique for accurate definition of the HLA-DQ types at the nucleotide level, eliminating the need for radioisotope as well as allele specific oligonucleotide probes and can be extended and applied to HLA-DR, -Dw DP typing. PMID:2576477

  12. The Development of a Novel, Validated, Rapid and Simple Method for the Detection of Sarcocystis fayeri in Horse Meat in the Sanitary Control Setting.

    PubMed

    Furukawa, Masato; Minegishi, Yasutaka; Izumiyama, Shinji; Yagita, Kenji; Mori, Hideto; Uemura, Taku; Etoh, Yoshiki; Maeda, Eriko; Sasaki, Mari; Ichinose, Kazuya; Harada, Seiya; Kamata, Yoichi; Otagiri, Masaki; Sugita-Konishi, Yoshiko; Ohnishi, Takahiro

    2016-01-01

    Sarcocystis fayeri (S. fayeri) is a newly identified causative agent of foodborne disease that is associated with the consumption of raw horse meat. The testing methods prescribed by the Ministry of Health, Labour and Welfare of Japan are time consuming and require the use of expensive equipment and a high level of technical expertise. Accordingly, these methods are not suitable for use in the routine sanitary control setting to prevent outbreaks of foodborne disease. In order to solve these problems, we have developed a new, rapid and simple testing method using LAMP, which takes only 1 hour to perform and which does not involve the use of any expensive equipment or expert techniques. For the validation of this method, an inter-laboratory study was performed among 5 institutes using 10 samples infected with various concentrations of S. fayeri. The results of the inter-laboratory study demonstrated that our LAMP method could detect S. fayeri at concentrations greater than 10(4) copies/g. Thus, this new method could be useful in screening for S. fayeri as a routine sanitary control procedure. PMID:27350431

  13. Simple and rapid method determination for metformin in human plasma using high performance liquid chromatography tandem mass spectrometry: application to pharmacokinetic studies.

    PubMed

    Marques, Marlice A Sípoli; Soares, Alcenir de Souza; Pinto, Olivia Woyames; Barroso, Pedro Tupinambá Werneck; Pinto, Douglas Pereira; Ferreira-Filho, Milton; Werneck-Barroso, Eduardo

    2007-06-01

    A rapid and simple method for quantitation of metformin (MET) in human plasma by HPLC-MS/MS was developed and validated. The sample preparation consists of plasma deproteinization using acetonitrile. The mobile phase consisted of water-acetonitrile and formic acid (55/45/0.048, v/v/%) and the run time was 3 min. A pursuit C(18) (100 mm x 2.0 mm i.d., 3 microm) column connected to a guard column MS-pursuit (0.20 mm x 0.20 mm i.d., 5 microm) was used. The range of the calibration curve was from 20 to 5000 ng/mL, the limit of quantitation being 20 ng/mL. The detection was performed on a mass spectrometer (ESI+), using metoprolol as internal standard. The calibration curves have r(2) values of 0.995 (CV=0.24%, n=10). The accuracy and precision were between 90.74 and 106.7% and coefficients of variations (CV) of 1.10 and 4.35%, respectively. The method was applied to determine the pharmacokinetic parameters: C(max) (1667.25 ng/mL) and T(max) (3.89 h). PMID:17331818

  14. A simple and rapid HPLC-DAD method for simultaneously monitoring the accumulation of alkaloids and precursors in different parts and different developmental stages of Catharanthus roseus plants.

    PubMed

    Pan, Qifang; Saiman, Mohd Zuwairi; Mustafa, Natali Rianika; Verpoorte, Robert; Tang, Kexuan

    2016-03-01

    A rapid and simple reversed phase liquid chromatographic system has been developed for simultaneous analysis of terpenoid indole alkaloids (TIAs) and their precursors. This method allowed separation of 11 compounds consisting of eight TIAs (ajmalicine, serpentine, catharanthine, vindoline, vindolinine, vincristine, vinblastine, and anhydrovinblastine) and three related precursors i.e., tryptophan, tryptamine and loganin. The system has been applied for screening the TIAs and precursors in Catharanthus roseus plant extracts. In this study, different organs i.e., flowers, leaves, stems, and roots of C. roseus were investigated. The results indicate that TIAs and precursor accumulation varies qualitatively and quantitatively in different organs of C. roseus. The precursors showed much lower levels than TIAs in all organs. Leaves and flowers accumulate higher level of vindoline, catharanthine and anhydrovinblastine while roots have higher level of ajmalicine, vindolinine and serpentine. Moreover, the alkaloid profiles of leaves harvested at different ages and different growth stages were studied. The results show that the levels of monoindole alkaloids decreased while bisindole alkaloids increased with leaf aging and upon plant growth. The HPLC method has been successfully applied to detect TIAs and precursors in different types of C. roseus samples to facilitate further study of the TIA pathway and its regulation in C. roseus plants. PMID:26854826

  15. Photocatalytic reduction of CO2 over Ag/TiO2 nanocomposites prepared with a simple and rapid silver mirror method.

    PubMed

    Yu, Bingcheng; Zhou, Yong; Li, Peng; Tu, Wenguang; Li, Ping; Tang, Lanqin; Ye, Jinhua; Zou, Zhigang

    2016-06-01

    The photocatalytic reduction of CO2 over Ag/TiO2 composites prepared with a simple silver mirror reaction method was investigated under UV-visible irradiation in both gas-phase (CO2 + water vapor) and aqueous solution (CO2-saturated NaHCO3 solution) systems. The as-prepared Ag/TiO2 nanocomposite exhibits efficient photocatalytic activity due to the surface plasmonic resonance and electron sink effect of the Ag component, which was found to be closely related to the size and loading amount of Ag. The rapid silver method is effective at curbing the size of Ag, so photocatalytic activity can be improved. Diverse organic chemical products were detected, including mainly methane and methanol as well as a small amount of C2 and C3 species such as acetaldehyde and acetone. Possible photocatalytic mechanisms were proposed. This artificial photosynthesis process may give a prosperous route to the removal of CO2 while simultaneously converting CO2 to valuable fuels based on highly efficient photocatalysts. PMID:27231820

  16. A simple and rapid method for direct determination of Al(III) based on the enhanced resonance Rayleigh scattering of hemin-functionalized graphene-Al(III) system

    NASA Astrophysics Data System (ADS)

    Ling, Yu; Chen, Ling Xiao; Dong, Jiang Xue; Li, Nian Bing; Luo, Hong Qun

    2016-03-01

    A novel method for direct determination of Al(III) by using hemin-functionalized graphene (H-GO) has been established based on the enhancement of resonance Rayleigh scattering (RRS) intensity. The characteristics of RRS spectra, the optimum reaction conditions, and the reaction mechanism have been investigated. In this experiment, the Al(III) would exist in sol-gel Al(OH)3 species under the condition of pH 5.9 in aqueous solutions. When H-GO existed in the solution, the sol-gel Al(OH)3 would react with H-GO and result in enhancement of RRS intensity, owing to the enhanced hydrophobicity of H-GO surface. Therefore, a simple and rapid sensor for Al(III) was developed. The increased intensity of RRS is directly proportional to the concentration of Al(III) in the range of 10 nM-6 μM, along with a detection limit of 0.87 nM. Moreover, the sensor has been applied to determination of Al(III) concentration in real water and aspirin tablet samples with satisfactory results. Therefore, the proposed method is promising as an effective means for selective and sensitive determination of Al(III).

  17. Specific ionic effect for simple and rapid colorimetric sensing assays of amino acids using gold nanoparticles modified with task-specific ionic liquid.

    PubMed

    Wu, Datong; Cai, Pengfei; Tao, Zhihao; Pan, Yuanjiang

    2016-01-01

    In this study, a novel task-specific ionic liquid functionalized gold nanoparticle (TSIL-GNP) was successfully prepared and applied in the recognition of amino acids. Particularly, the surface of GNP was modified with the ionic liquid containing carbamido and ester group via thiol, which was characterized by Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). The stability of this material in aqueous solution improves apparently and can remain unchanged for more than three months. The effect of pH was also discussed in this study. Attractive ionic interaction would effectively weaken intensity of the covalent coupling between the metal ion and the functional groups of amino acids. Thus, TSIL-GNP was successfully applied to recognizing serine, aspartic acid, lysine, arginine, and histidine in the presence of Cu(2+) through distinctive color changes. Suspension would be generated once a spot of cysteine was added into the GNPs solution. Results indicated that it had a good linear relationship between extinction coefficients and concentration of amino acids in a wide range of 10(-3)-10(-6) M. Moreover, the proposed strategy was successfully used to analyze the histidine in urinary samples. In brief, TSIL-GNP is a suitable substrate for discrimination of five amino acids in a rapid and simple way without sophisticated instruments. PMID:26703268

  18. Development of a rapid, simple and sensitive HPLC-FLD method for determination of rhodamine B in chili-containing products.

    PubMed

    Qi, Ping; Lin, Zhihao; Li, Jiaxu; Wang, ChengLong; Meng, WeiWei; Hong, Hong; Zhang, Xuewu

    2014-12-01

    In this work, a simple, rapid and sensitive analytical method for the determination of rhodamine B in chili-containing foodstuffs is described. The dye is extracted from samples with methanol and analysed without further cleanup procedure by high-performance liquid chromatography (HPLC) coupled to fluorescence detection (FLD). The influence of matrix fluorescent compounds (capsaicin and dihydrocapsaicin) on the analysis was overcome by the optimisation of mobile-phase composition. The limit of determination (LOD) and limit of quantification (LOQ) were 3.7 and 10 μg/kg, respectively. Validation data show a good repeatability and within-lab reproducibility with relative standard deviations <10%. The overall recoveries are in the range of 98-103% in chili powder and in the range of 87-100% in chili oil depending on the concentration of rhodamine B in foodstuffs. This method is suitable for the routine analysis of rhodamine B due to its sensitivity, simplicity, reasonable time and cost. PMID:24996311

  19. Molecularly imprinted polymer cartridges coupled on-line with high performance liquid chromatography for simple and rapid analysis of human insulin in plasma and pharmaceutical formulations.

    PubMed

    Moein, Mohammad Mahdi; Javanbakht, Mehran; Akbari-adergani, Behrouz

    2014-04-01

    In this paper, a novel method is described for automated determination of human insulin in biological fluids using principle of sequential injection on a molecularly imprinted solid-phase extraction (MISPE) cartridge as a sample clean-up technique combined with high performance liquid chromatography (HPLC). The water-compatible molecularly imprinted polymers (MIPs) were prepared using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross-linker, chloroform as a porogen and insulin as a template molecule. The imprinted polymers were then employed as the solid-phase extraction sorbent for on-line extraction of insulin from human plasma samples. To achieve the best condition, influential parameters on the extraction efficiency were thoroughly investigated. Rapid and simple analysis of the hormone was successfully accomplished through the good selectivity of the prepared sorbent coupled with HPLC. Limits of detection (LOD) and quantification (LOQ) of 0.2 ng mL(-1), 0.7 ng mL(-1), and 0.03 ng mL(-1), 0.1 ng mL(-1) were obtained in plasma and urine respectively. The obtained data exhibited the great recoveries for extraction of insulin from human plasma and pharmaceutical samples, higher than 87%. PMID:24607106

  20. Simple and rapid method for detection of nitrate reductase activity of Mycobacterium tuberculosis and Mycobacterium canettii grown in the Bactec MGIT960 system.

    PubMed

    Goh, Khye Seng; Rastogi, Nalin

    2010-05-01

    Mycobacterium tuberculosis reduces nitrate very strongly as compared to Mycobacterium bovis and M. bovis BCG. Nitrate reductase, in conjunction with niacin accumulation, constitutes one of the major biochemical tests used in clinical microbiology laboratories to differentiate M. tuberculosis from other members of the M. tuberculosis complex, as well as nontuberculous Mycobacteria. Determination of nitrate reductase activity is currently performed using cultures grown on solid media with a slow detection time and the need for large quantities of bacilli, as otherwise the test is not reliable. Hereby, we propose a nitrate reduction test coupled to Bactec MGIT960 system as a simple, rapid and economic method with a total gain of time of about 3 to 4weeks over the conventional solid medium. In our study, almost all the M. tuberculosis and Mycobacterium canettii strains gave a strongly positive nitrate reductase result within 1day of positive detection by the MGIT960 system. In contrast, M. bovis, M. bovis BCG and M. africanum strains remained negative even after 14days of incubation. The possibility to detect nitrate reductase within 1 to 3days of a positive culture using MGIT960 opens new perspectives with the possibility of confirming M. tuberculosis - starting directly from pathological specimens. PMID:20298726

  1. Evaluation of Paracheck-PfTM rapid malaria diagnostic test for the diagnosis of malaria among HIV-positive patients in Ibadan, south-western Nigeria

    PubMed Central

    Falade, C O; Adesina-Adewole, B; Dada-Adegbola, H O; Ajayi, I O; Akinyemi, J O; Ademowo, O G; Adewole, I F; Kanki, P

    2013-01-01

    Febrile illnesses occur frequently among HIV positive patients and these are often treated presumptively as malaria in endemic areas. Parasite-based diagnosis of malaria will eliminate unnecessary treatment, reduce drug–drug interactions and the chances for the emergence of drug resistant Plasmodium. We evaluated finger prick blood samples from 387 people living with HIV (PLWHIV) and suspected of having malaria by expert microscopy and Paracheck-PfTM – a histidine-rich protein-II based malaria rapid diagnostic test. The study was conducted at the PEPFAR supported AIDS Prevention Initiative in Nigeria (APIN) Clinic of the University College Hospital Ibadan, southwest Nigeria. Outcome parameters were prevalence of malaria parasitemia, sensitivity and specificity of Paracheck-Pf as well as the positive and negative predictive values for Paracheck-Pf using microscopy of Giemsa-stained blood film as gold standard. Malaria parasites were detected in 19.1% (74/387) of enrollees by microscopy and 19.3% (74/383) by Paracheck-Pf. Geometric mean parasite density was 501/μl (range 39–749 202/μl). Sensitivity and specificity of Paracheck-Pf at all parasite densities were 55.4% and 89.3% while corresponding figures at parasite densities ≧200/μl were 90.9% and 90.3%. Sensitivity and specificity at parasite densities ≧500/μl was 97.6% and 90.3%. Positive and negative predictive values for parasite density ≧200/μl were 55.4% and 98.7%, respectively. Paracheck-pf was found to be a useful malaria diagnostic tool at parasite densities ≧200/μl facilitating appropriate clinical management. PMID:23683333

  2. Rapid, low-cost and instrument-free CD4+ cell counting for HIV diagnostics in resource-poor settings.

    PubMed

    Glynn, Macdara T; Kinahan, David J; Ducrée, Jens

    2014-08-01

    We present a novel, user-friendly and widely autonomous point-of-care diagnostic to enable HIV monitoring in resource-poor regions where the current pandemic is most prevalent. To specifically isolate magnetically tagged CD4+ cells directly from patient blood, the low-cost and disposable microfluidic chip operates by dual-force CD4+ cell magnetophoresis; whereby the interplay of flow and magnetic fields governs the trajectory of target cells depending on whether the cell binds to a magnetic microbead. Instrument-free pumping is implemented by a finger-actuated elastic membrane; tagged beads are laterally deflected by a small and re-useable permanent magnet. The single-depth and monolithic microfluidic structure can easily be fabricated in a single casting step. After their magnetophoretic isolation from whole blood, estimation of CD4+ cell concentrations is then measured by bright-field inspection of the capture chamber. In addition, an optional fluorescence measurement can be used for confirmation of the bright-field result if required. On-chip CD4+ estimation produces a linear response over the full range of medically relevant CD4+ cell concentrations. Our technology combines high-efficiency capture (93.0 ± 3.3%) and cell enumeration. PMID:24911165

  3. HIV-1 protein gp120 rapidly impairs memory in chicks by interrupting the glutamate-glutamine cycle.

    PubMed

    Fernandes, S P; Edwards, T M; Ng, K T; Robinson, S R

    2007-01-01

    Learning and memory impairments are frequently observed in patients suffering from AIDS Dementia Complex (ADC). These effects have been linked to the presence of gp120, an HIV viral coat glycoprotein. The present study investigated the possibility that gp120 prevents the uptake of extracellular glutamate by astrocytes, leading to an interruption of the glutamate-glutamine cycle and a subsequent impairment of memory. Ten microliters of 10nM gp120 was bilaterally injected into the region of the intermediate medial mesopallium of day-old chicks at various times before, or after, training using a single-trial passive avoidance task. Gp120 was found to significantly impair memory retention when injected 10-40 min after training. Memory impairments were evident within 5 min of gp120 administration and remained evident 24h later. Further, the amnestic effect of gp120 could be overcome with glutamine or with precursors of glutamate synthesis, but only weakly by glutamate. These results support the conclusion that the amnestic effect of gp120 is due to an impaired uptake of glutamate by astrocytes and a subsequent interruption of glutamine supply to neurones. The data indicate that the glutamate-glutamine cycle may be a useful therapeutic target in the treatment of ADC. PMID:16714124

  4. Rapid Urine LAM Testing Improves Diagnosis of Expectorated Smear-Negative Pulmonary Tuberculosis in an HIV-endemic Region

    PubMed Central

    Drain, Paul K.; Gounder, Lilishia; Sahid, Faieza; Moosa, Mahomed-Yunus S.

    2016-01-01

    We sought to determine if urine lipoarabinomannan (LAM) would improve diagnosis of pulmonary TB. We enrolled consecutive adults presenting with ≥2 TB-related symptoms, obtained one induced sputum sample for smear microscopy (AFB) and mycobacterial culture, and performed urine LAM testing (DetermineTM TB LAM, Alere). We used culture-confirmed pulmonary TB as the gold standard, and compared accuracy with area under receiver operating characteristic curves (AUROC). Among 90 participants, 82 of 88 tested (93%) were HIV-infected with a median CD4 168/mm3 (IQR 89–256/mm3). Diagnostic sensitivities of urine LAM and sputum AFB were 42.1% (95% CI 29.1–55.9%) and 21.1% (95% CI 11.4–33.9%), and increased to 52.6% (95% CI 39.0–66.0%) when combined. Sensitivity of LAM increased significantly among participants with a lower Karnofsky Performance score, anemia, hypoalbuminemia, and higher C-reactive protein. Combining LAM with AFB had an AUROC = 0.68 (95% CI 0.59–0.77), significantly better than AFB alone (AUROC=0.58; 95% CI 0.51–0.64). The combination of LAM and AFB was significantly better than AFB alone among patients with Karnofsky Performance score ≤90, hemoglobin ≤10 g/dL, albumin ≤25 g/L, C-reactive protein ≥25 mg/L, or CD4 <200/mm3. Urine LAM testing may be most beneficial among patients with functional impairment, elevated inflammatory markers, or greater immunosuppression. PMID:26865526

  5. Acute retroviral syndrome and high baseline viral load are predictors of rapid HIV progression among untreated Argentinean seroconverters

    PubMed Central

    2011-01-01

    Background Diagnosis of primary HIV infection (PHI) has important clinical and public health implications. HAART initiation at this stage remains controversial. Methods Our objective was to identify predictors of disease progression among Argentinean seroconverters during the first year of infection, within a multicentre registry of PHI-patients diagnosed between 1997 and 2008. Cox regression was used to analyze predictors of progression (LT-CD4 < 350 cells/mm3, B, C events or death) at 12 months among untreated patients. Results Among 134 subjects, 74% presented with acute retroviral syndrome (ARS). Seven opportunistic infections (one death), nine B events, and 10 non-AIDS defining serious events were observed. Among the 92 untreated patients, 24 (26%) progressed at 12 months versus three (7%) in the treated group (p = 0.01). The 12-month progression rate among untreated patients with ARS was 34% (95% CI 22.5-46.3) versus 13% (95% CI 1.1-24.7) in asymptomatic patients (p = 0.04). In univariate analysis, ARS, baseline LT-CD4 < 350 cells/mm3, and baseline and six-month viral load (VL) > 100,000 copies/mL were associated with progression. In multivariate analysis, only ARS and baseline VL > 100,000 copies/mL remained independently associated; HR: 8.44 (95% CI 0.97-73.42) and 9.44 (95% CI 1.38-64.68), respectively. Conclusions In Argentina, PHI is associated with significant morbidity. HAART should be considered in PHI patients with ARS and high baseline VL to prevent disease progression. PMID:21831310

  6. Simple and rapid yeast reporter bioassay for dioxin screening: evaluation of the dioxin-like compounds in industrial and municipal waste incineration plants.

    PubMed

    Kawanishi, Masanobu; Ohnisi, Kana; Takigami, Hidetaka; Yagi, Takashi

    2013-05-01

    The CROMIS AhR kit, a simple and rapid yeast bioassay kit, was developed and used to detect dioxins and dioxin-like compounds in 20 gas and solid samples collected from refuse incineration plants in Japan. The World Health Organization toxic equivalent (WHO-TEQ) values of the samples were also calculated using high-resolution gas chromatography/high--resolution mass spectrometry. The WHO-TEQ values of the samples varied greatly, ranging from 0.0021-6.3 ng/g to 0.00013-16 ng/m(3)N (normal cubic meter) in the solid and gas samples, respectively. 2,3,4,7,8-Pentachlorodibenzofuran (23478-PeCDF) and 1,2,3,7,8-pentachlorodibenzo-p-dioxin (12378-PeCDD) were the major contributors to the samples' WHO-TEQ values. The yeast in the bioassay responded to these congeners, and the EC50 values of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2378-TeCDD), 12378-PeCDD, and 2,3,4,7,8-PeCDF were 490, 560, and 590 nM, respectively. The incinerator samples were subjected to the bioassay to obtain 2378-TeCDD equivalent values (CROMIS-TEQ values). The CROMIS-TEQ values of the solid and gas samples ranged from 0.0019 to 5.64 ng/g and from 0.14 to 20 ng/m(3)N, respectively. The CROMIS-TEQ and WHO-TEQ values displayed good correlations (r (2) = 0.94 and 0.95 in the solid and gas samples, respectively), except for those of the samples with low dioxin concentrations (below the Japanese emission standards). Therefore, the CROMIS AhR kit is a useful tool for the initial screening of samples containing dioxin-like compounds. PMID:23054780

  7. A simple and rapid method for measuring α-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector

    PubMed Central

    Jia, Xiaochen; Kang, Jian

    2016-01-01

    The interconversion of hexose-6-phosphate and hexose-1-phosphate can be directly analyzed by high-performance anion-exchange chromatography coupled with an electrochemical detector (HPAEC-PAD). Thus, this method can be used to measure the activities of N-acetylglucosamine-phosphate mutase (AGM), glucosamine-phosphate mutase (GlmM) and phosphoglucomutase (PGM), which are the members of α-D-phosphohexomutases superfamily. The detection limits were extremely low as 2.747 pmol, 1.365 pmol, 0.512 pmol, 0.415 pmol, 1.486 pmol and 0.868 pmol for N-acetylglucosamine-1-phosphate (GlcNAc-1-P), N-acetylglucosamine-6-phosphate (GlcNAc-6-P), glucosamine-1-phosphate (GlcN-1-P), glucosamine-6-phosphate (GlcN-6-P), glucose-1-phosphate (Glc-1-P) and glucose-6-phosphate (Glc-6-P), respectively. By employing HPAEC-PAD, activities of AtAGM (AGM from Arabidopsis thaliana) on these six phosphohexoses can be detected. The Km of AtAGM on Glc-1-P determined by HPAEC-PAD was 679.18 ± 156.40 µM, which is comparable with the Km of 707.09 ± 170.36 µM detected by traditional coupled assay. Moreover, the activity of MtGlmM (GlmM from Mycobacterium tuberculosis) on GlcN-6-P tested by HPAEC-PAD was 7493.40 ± 309.12 nmol∕min ⋅ mg, which is much higher than 288.97 ± 35.28 nmol∕min ⋅ mg obtained by the traditional coupled assay. Accordingly, HPAEC-PAD is a more rapid and simple method than the traditional coupled assays given its high specificity and sensitivity, and will certainly bring convenience to further research of α-D-phosphohexomutases. PMID:26788420

  8. A simple and rapid method for measuring α-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector.

    PubMed

    Jia, Xiaochen; Kang, Jian; Yin, Heng

    2016-01-01

    The interconversion of hexose-6-phosphate and hexose-1-phosphate can be directly analyzed by high-performance anion-exchange chromatography coupled with an electrochemical detector (HPAEC-PAD). Thus, this method can be used to measure the activities of N-acetylglucosamine-phosphate mutase (AGM), glucosamine-phosphate mutase (GlmM) and phosphoglucomutase (PGM), which are the members of α-D-phosphohexomutases superfamily. The detection limits were extremely low as 2.747 pmol, 1.365 pmol, 0.512 pmol, 0.415 pmol, 1.486 pmol and 0.868 pmol for N-acetylglucosamine-1-phosphate (GlcNAc-1-P), N-acetylglucosamine-6-phosphate (GlcNAc-6-P), glucosamine-1-phosphate (GlcN-1-P), glucosamine-6-phosphate (GlcN-6-P), glucose-1-phosphate (Glc-1-P) and glucose-6-phosphate (Glc-6-P), respectively. By employing HPAEC-PAD, activities of AtAGM (AGM from Arabidopsis thaliana) on these six phosphohexoses can be detected. The Km of AtAGM on Glc-1-P determined by HPAEC-PAD was 679.18 ± 156.40 µM, which is comparable with the Km of 707.09 ± 170.36 µM detected by traditional coupled assay. Moreover, the activity of MtGlmM (GlmM from Mycobacterium tuberculosis) on GlcN-6-P tested by HPAEC-PAD was 7493.40 ± 309.12 nmol∕min ⋅ mg, which is much higher than 288.97 ± 35.28 nmol∕min ⋅ mg obtained by the traditional coupled assay. Accordingly, HPAEC-PAD is a more rapid and simple method than the traditional coupled assays given its high specificity and sensitivity, and will certainly bring convenience to further research of α-D-phosphohexomutases. PMID:26788420

  9. Size-exclusion HPLC provides a simple, rapid, and versatile alternative method for quality control of vaccines by characterizing the assembly of antigens.

    PubMed

    Yang, Yanli; Li, Hao; Li, Zhengjun; Zhang, Yan; Zhang, Songping; Chen, Yi; Yu, Mengran; Ma, Guanghui; Su, Zhiguo

    2015-02-25

    The assembly of antigen structure is often crucial to the potency of vaccines. Currently adopted methods like animal testing and ultracentrifugation take long time and are difficult to automate for multiple samples. Here we develop a size-exclusion high-performance liquid chromatography (SE-HPLC) method to characterize the assembly of antigen structure during both manufacturing process and storage. Three important vaccine antigens including inactivated foot and mouth disease virus (FMDV), which is a virus vaccine; and two virus-like particles (VLPs) vaccines involving hepatitis B core antigen (HBcAg) VLPs, and hepatitis B surface antigen (HBsAg) VLPs, were successfully analyzed using commercially available TSK gel columns with pore size above 45nm. Combined with other analytical methods including SDS-PAGE, dynamic light scattering, wavelength scan, and multi-angle laser light scattering, the SE-HPLC method was proven to be a simple, rapid, and reliable tool for antigen particles assembly analysis. Specifically, for FMDV whole virus particle, SE-HPLC was used to analyze 146S content in vaccine preparations and the thermal dissociation of the 146S. For HBcAg-VLPs that are expressed in recombinant Escherichia coli, its expression level during cell culture process was quantitatively monitored by SE-HPLC. The SE-HPLC also showed applicability for quality check of HBsAg vaccine preparations by monitoring the product consistency of different lot number and the product stability during storage. Results shown in this work clearly demonstrated that SE-HPLC method has potential as a versatile alternative technology for control of the final product by both manufacturers and the regulatory agencies. PMID:25604799

  10. Sorptive thin film microextraction followed by direct solid state spectrofluorimetry: A simple, rapid and sensitive method for determination of carvedilol in human plasma.

    PubMed

    Karimi, Shima; Talebpour, Zahra; Adib, Noushin

    2016-06-14

    A poly acrylate-ethylene glycol (PA-EG) thin film is introduced for the first time as a novel polar sorbent for sorptive extraction method coupled directly to solid-state spectrofluorimetry without the necessity of a desorption step. The structure, polarity, fluorescence property and extraction performance of the developed thin film were investigated systematically. Carvedilol was used as the model analyte to evaluate the proposed method. The entire procedure involved one-step extraction of carvedilol from plasma using PA-EG thin film sorptive phase without protein precipitation. Extraction variables were studied in order to establish the best experimental conditions. Optimum extraction conditions were the followings: stirring speed of 1000 rpm, pH of 6.8, extraction temperature of 60 °C, and extraction time of 60 min. Under optimal conditions, extraction of carvedilol was carried out in spiked human plasma; and the linear range of calibration curve was 15-300 ng mL(-1) with regression coefficient of 0.998. Limit of detection (LOD) for the method was 4.5 ng mL(-1). The intra- and inter-day accuracy and precision of the proposed method were evaluated in plasma sample spiked with three concentration levels of carvedilol; yielding a recovery of 91-112% and relative standard deviation of less than 8%, respectively. The established procedure was successfully applied for quantification of carvedilol in plasma sample of a volunteer patient. The developed PA-EG thin film sorptive phase followed by solid-state spectrofluorimetric method provides a simple, rapid and sensitive approach for the analysis of carvedilol in human plasma. PMID:27181643

  11. A simple, rapid and reliable liquid chromatography-mass spectrometry method for determination of methotrexate in human plasma and its application to therapeutic drug monitoring.

    PubMed

    Wu, Dan; Wang, Yixuan; Sun, Yan; Ouyang, Nian; Qian, Jun

    2015-08-01

    A simple, rapid and reliable liquid chromatography-electrospray ionization tandem mass spectrometry method was established and validated for the determination of methotrexate in human plasma. After a straightforward protein precipitation by acetonitrile-water (70:30, v/v), methotrexate (MTX) and p-aminoacetophenone (used as internal standard, IS) were separated on a Column C18 column (50 × 2.1 mm, 3 µm; Column Technology, Fremont, CA, USA) using a gradient elution with mobile phase of acetonitrile and 0.03% acetic acid aqueous solution at a flow rate of 0.5 mL/min. The total chromatographic runtime was 5 min for each injection. Quantification detection was performed in a triple-quadruple tandem mass spectrometer under positive mode monitoring the following mass transitions: m/z 455.3 → 308.3 for MTX and m/z 136.1 → 94.4 for IS. The calibration curve was linear over the range of 0.05-25.0 µmol/L with a lower limit of quantification of 0.05 µmol/L. The intra- and interday precisions were <5.2%, the accuracy varied from -4.1 to 4.5%. The recovery was >94%. The LC-MS/MS method showed an excellent agreement with the existing HPLC-UV method using Passing-Bablok regression and Bland-Altman difference plot analysis. The validated LC-MS/MS can be successfully applied to the routine therapeutic drug monitoring of MTX in clinical laboratories. PMID:25641007

  12. A rapid and simple method for the determination of 3,4-dihydroxyphenylacetic acid, norepinephrine, dopamine, and serotonin in mouse brain homogenate by HPLC with fluorimetric detection.

    PubMed

    De Benedetto, Giuseppe Egidio; Fico, Daniela; Pennetta, Antonio; Malitesta, Cosimino; Nicolardi, Giuseppe; Lofrumento, Dario Domenico; De Nuccio, Francesco; La Pesa, Velia

    2014-09-01

    A fast and simple isocratic high-performance liquid chromatography method for the determination of 3,4-dihydroxyphenylacetic acid (DOPAC), norepinephrine (NE), dopamine (DA), and serotonin (5-HT) in homogenate samples of mouse striatum employing the direct fluorescence of the neurotransmitters is described. The method has been optimized and validated. The analytes were separated in 15min on a reversed-phase column (C18) with acetate buffer (pH 4.0, 12mM)-methanol (86:14, v/v) as mobile phase; the flow rate was 1ml/min. The fluorescence measurements were carried out at 320nm with excitation at 279nm. The calibration curve for DA was linear up to about 2.5μg/ml, with a coefficient of determination (r(2)) of 0.9995 with a lower limit of quantification of 0.031μg/ml. Since the procedure does not involve sample pre-purification or derivatisation, the recovery ranged from 97% to 102% and relative standard deviation (RSD) was better than 2.9%, the use of the internal standard is not mandatory, further simplifying the method. Similar performance was obtained for the other analytes. As a result, thanks to its simplicity, rapidity and adequate working range, the method can be used for the determination of 3,4-dihydroxyphenylacetic acid, dopamine, norepinephrine and serotonin in animal tissues. An experimental 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson-like disease has been used to demonstrate the method is fit-for-purpose. PMID:24971521

  13. Simple Machines Made Simple.

    ERIC Educational Resources Information Center

    St. Andre, Ralph E.

    Simple machines have become a lost point of study in elementary schools as teachers continue to have more material to cover. This manual provides hands-on, cooperative learning activities for grades three through eight concerning the six simple machines: wheel and axle, inclined plane, screw, pulley, wedge, and lever. Most activities can be…

  14. Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody

    PubMed Central

    Sainsbury, Frank; Sack, Markus; Stadlmann, Johannes; Quendler, Heribert; Fischer, Rainer; Lomonossoff, George P.

    2010-01-01

    Background The capacity of plants and plant cells to produce large amounts of recombinant protein has been well established. Due to advantages in terms of speed and yield, attention has recently turned towards the use of transient expression systems, including viral vectors, to produce proteins of pharmaceutical interest in plants. However, the effects of such high level expression from viral vectors and concomitant effects on host cells may affect the quality of the recombinant product. Methodology/Principal Findings To assess the quality of antibodies transiently expressed to high levels in plants, we have expressed and characterised the human anti-HIV monoclonal antibody, 2G12, using both replicating and non-replicating systems based on deleted versions of Cowpea mosaic virus (CPMV) RNA-2. The highest yield (approximately 100 mg/kg wet weight leaf tissue) of affinity purified 2G12 was obtained when the non-replicating CPMV-HT system was used and the antibody was retained in the endoplasmic reticulum (ER). Glycan analysis by mass-spectrometry showed that the glycosylation pattern was determined exclusively by whether the antibody was retained in the ER and did not depend on whether a replicating or non-replicating system was used. Characterisation of the binding and neutralisation properties of all the purified 2G12 variants from plants showed that these were generally similar to those of the Chinese hamster ovary (CHO) cell-produced 2G12. Conclusions Overall, the results demonstrate that replicating and non-replicating CPMV-based vectors are able to direct the production of a recombinant IgG similar in activity to the CHO-produced control. Thus, a complex recombinant protein was produced with no apparent effect on its biochemical properties using either high-level expression or viral replication. The speed with which a recombinant pharmaceutical with excellent biochemical characteristics can be produced transiently in plants makes CPMV-based expression vectors

  15. Pyrosequencing-based validation of a simple cell-suspension polymerase chain reaction assay for Campylobacter... of high-processivity polymerase with novel internal amplification controls for rapid and specific detection.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although Campylobacter is an important food-borne human pathogen, there remains a lack of molecular diagnostic assays that are simple to use, cost-effective, and provide rapid results in research, clinical, or regulatory laboratories. Of the numerous Campylobacter assays that do exist, to our knowl...

  16. BST-2 is rapidly down-regulated from the cell surface by the HIV-1 protein Vpu: evidence for a post-ER mechanism of Vpu-action

    PubMed Central

    Skasko, Mark; Tokarev, Andrey; Chen, Cheng-Chang; Fischer, Wolfgang B.; Pillai, Satish K.; Guatelli, John

    2011-01-01

    Recent evidence suggests that transmembrane domain (TMD) interactions are essential for HIV-1 Vpu-mediated antagonism of the restriction factor BST-2/tetherin. We made Vpu TMD mutants to study the mechanism of BST-2 antagonism. Vpu-I17A, -A18F, - W22L, and -S23L co-localized with BST-2 within endosomal membranes while effectively enhancing virion release and down-regulating surface BST-2. However, Vpu-A18H was confined to an endoplasmic reticulum (ER)-like distribution, resulting in impaired down-regulation of BST-2 and reduced virion release. Brefeldin A confined wild type Vpu to the ER, resulting in a similarly impaired phenotype, as did the addition of a C-terminal ER-retention signal to Vpu. We determined the half-life of cell-surface BST-2 to be ~8 hours, whereas Vpu mediated an ~80% reduction of surface BST-2 within 6 hours, suggesting that TMD interactions between Vpu and BST-2 occur within post-ER membranes to directly and rapidly remove BST-2 from the cell surface and relieve restricted virion release. PMID:21237475

  17. Key gp120 Glycans Pose Roadblocks to the Rapid Development of VRC01-Class Antibodies in an HIV-1-Infected Chinese Donor.

    PubMed

    Kong, Leopold; Ju, Bin; Chen, Yajing; He, Linling; Ren, Li; Liu, Jiandong; Hong, Kunxue; Su, Bin; Wang, Zheng; Ozorowski, Gabriel; Ji, Xiaolin; Hua, Yuanzi; Chen, Yanli; Deller, Marc C; Hao, Yanling; Feng, Yi; Garces, Fernando; Wilson, Richard; Dai, Kaifan; O'Dell, Sijy; McKee, Krisha; Mascola, John R; Ward, Andrew B; Wyatt, Richard T; Li, Yuxing; Wilson, Ian A; Zhu, Jiang; Shao, Yiming

    2016-04-19

    VRC01-class antibodies neutralize diverse HIV-1 strains by targeting the conserved CD4-binding site. Despite extensive investigations, crucial events in the early stage of VRC01 development remain elusive. We demonstrated how VRC01-class antibodies emerged in a Chinese donor by antigen-specific single B cell sorting, structural and functional studies, and longitudinal antibody and virus repertoire analyses. A monoclonal antibody DRVIA7 with modest neutralizing breadth was isolated that displayed a subset of VRC01 signatures. X-ray and EM structures revealed a VRC01-like angle of approach, but less favorable interactions between the DRVIA7 light-chain CDR1 and the N terminus with N276 and V5 glycans of gp120. Although the DRVIA7 lineage was unable to acquire broad neutralization, longitudinal analysis revealed a repertoire-encoded VRC01 light-chain CDR3 signature and VRC01-like neutralizing heavy-chain precursors that rapidly matured within 2 years. Thus, light chain accommodation of the glycan shield should be taken into account in vaccine design targeting this conserved site of vulnerability. PMID:27067056

  18. A New Method for Rapid Screening of End-Point PCR Products: Application to Single Genome Amplified HIV and SIV Envelope Amplicons.

    PubMed

    Houzet, Laurent; Deleage, Claire; Satie, Anne-Pascale; Merlande, Laetitia; Mahe, Dominique; Dejucq-Rainsford, Nathalie

    2015-01-01

    PCR is the most widely applied technique for large scale screening of bacterial clones, mouse genotypes, virus genomes etc. A drawback of large PCR screening is that amplicon analysis is usually performed using gel electrophoresis, a step that is very labor intensive, tedious and chemical waste generating. Single genome amplification (SGA) is used to characterize the diversity and evolutionary dynamics of virus populations within infected hosts. SGA is based on the isolation of single template molecule using limiting dilution followed by nested PCR amplification and requires the analysis of hundreds of reactions per sample, making large scale SGA studies very challenging. Here we present a novel approach entitled Long Amplicon Melt Profiling (LAMP) based on the analysis of the melting profile of the PCR reactions using SYBR Green and/or EvaGreen fluorescent dyes. The LAMP method represents an attractive alternative to gel electrophoresis and enables the quick discrimination of positive reactions. We validate LAMP for SIV and HIV env-SGA, in 96- and 384-well plate formats. Because the melt profiling allows the screening of several thousands of PCR reactions in a cost-effective, rapid and robust way, we believe it will greatly facilitate any large scale PCR screening. PMID:26053379

  19. HIV Risk among MSM in Senegal: A Qualitative Rapid Assessment of the Impact of Enforcing Laws That Criminalize Same Sex Practices

    PubMed Central

    Poteat, Tonia; Diouf, Daouda; Drame, Fatou Maria; Ndaw, Marieme; Traore, Cheikh; Dhaliwal, Mandeep; Beyrer, Chris; Baral, Stefan

    2011-01-01

    Men who have sex with men (MSM) are at high risk for HIV in Senegal, with a prevalence of 21.5%. In December 2008, nine male HIV prevention workers were imprisoned for “acts against nature” prohibited by Senegalese law. This qualitative study assessed the impact of these arrests on HIV prevention efforts. A purposive sample of MSM in six regions of Senegal was recruited by network referral. 26 in-depth interviews (IDIs) and 6 focus group discussions (FGDs) were conducted in July–August 2009. 14 key informants were also interviewed. All participants reported pervasive fear and hiding among MSM as a result of the December 2008 arrests and publicity. Service providers suspended HIV prevention work with MSM out of fear for their own safety. Those who continued to provide services noticed a sharp decline in MSM participation. An effective response to the HIV epidemic in Senegal should include active work to decrease enforcement of this law. PMID:22194906

  20. The Impact of Rapid HIV Home Test Use with Sexual Partners on Subsequent Sexual Behavior among Men Who Have Sex with Men

    PubMed Central

    Balán, Iván C.; Carballo-Diéguez, Alex; Frasca, Timothy; Dolezal, Curtis; Ibitoye, Mobolaji

    2013-01-01

    This study explores the sexual behavior 27 men who have sex with men (MSM) who regularly engage in unprotected anal intercourse (UAI), in the context of HIV home test (HT) use with potential sex partners. Participants were given 16 HT kits to use over three months. Among 40 sexual occasions following HIV-negative HT results, there were 25 UAI occasions (16 based on not typically using condoms and nine on HT results), 15 occasions in which condoms were used, and three in which sex did not occur. In the seven occasions where a potential partner received HIV-positive HT results, the sexual encounter ended. Almost all participants encountered potential partners who refused HT. Over half of these participants ended sexual encounters when HT was refused, perceiving these partners as HIV-positive or too high risk. Some participants reported that HT use heightened their awareness of HIV risk and their commitment to reducing it. PMID:23657758

  1. HIV Associated Opportunistic Pneumonias.

    PubMed

    Ismail, T; Lee, C

    2011-03-01

    Opportunistic pneumonias are major causes of morbidity and mortality in HIV infected individuals. The majority of new HIV infections in Malaysia are adults aged 20 to 39 years old and many are unaware of their HIV status until they present with an opportunistic infection. HIV associated opportunistic pneumonias can progress rapidly without appropriate therapy. Therefore a proper diagnostic evaluation is vital and prompt empiric treatment of the suspected diagnosis should be commenced while waiting for the results of the diagnostic studies. Tuberculosis, Pneumocystis pneumonia (PCP) and recurrent bacterial pneumonias are common causes of AIDS-defining diseases and are discussed in this article. PMID:23765154

  2. Development of a definition for Rapid Progression (RP) of renal function in HIV-positive persons: the D:A:D study

    PubMed Central

    2014-01-01

    Background No consensus exists on how to define abnormally rapid deterioration in renal function (Rapid Progression, RP). We developed an operational definition of RP in HIV-positive persons with baseline estimated glomerular filtration rate (eGFR) >90 ml/min/1.73 m2 (using Cockcroft Gault) in the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D) study from 2004 to 2011. Methods Two definitions were evaluated; RP definition A: An average eGFR decline (slope) ≥5 ml/min/1.73 m2/year over four years of follow-up with ≥3 eGFR measurements/year, last eGFR <90 ml/min/1.73 m2 and an absolute decline ≥5 ml/min/1.73 m2/year in two consecutive years. RP definition B: An absolute annual decline ≥5 ml/min/1.73 m2/year in each year and last eGFR <90 ml/min/1.73 m2. Sensitivity analyses were performed considering two and three years’ follow-up. The percentage with and without RP who went on to subsequently develop incident chronic kidney disease (CKD; 2 consecutive eGFRs <60 ml/min/1.73 m2 and 3 months apart) was calculated. Results 22,603 individuals had baseline eGFR ≥90 ml/min/1.73 m2. 108/3655 (3.0%) individuals with ≥4 years’ follow-up and ≥3 measurements/year experienced RP under definition A; similar proportions were observed when considering follow-up periods of three (n=195/6375; 3.1%) and two years (n=355/10756; 3.3%). In contrast under RP definition B, greater proportions experienced RP when considering two years (n=476/10756; 4.4%) instead of three (n=48/6375; 0.8%) or four (n=15/3655; 0.4%) years’ follow-up. For RP definition A, 13 (12%) individuals who experienced RP progressed to CKD, and only (21) 0.6% of those without RP progressed to CKD (sensitivity 38.2% and specificity 97.4%); whereas for RP definition B, fewer RP individuals progressed to CKD. Conclusions Our results suggest using three years’ follow-up and at least two eGFR measurements per year is most appropriate for a RP definition, as it allows inclusion of a reasonable

  3. A simplified and less expensive strategy for confirming anti HIV-1 screening results in a diagnostic laboratory in Lubumbashi, Zaire.

    PubMed

    Laleman, G; Kambale, M; Van Kerckhoven, I; Kapila, N; Konde, M; Selemani, U; Piot, P; van der Groen, G

    1991-12-01

    The conventional algorithm for HIV testing based on the confirmation of all positive anti-HIV screening reactions by Western blot (WB) is too expensive for developing countries. We investigated the validity of confirming positive screening assay reactions by a second screening test, limiting the use of the supplemental assay to the discrepant test results (algorithm 3), or screening all sera with 2 different assays and retesting all discrepant results by a supplemental assay (algorithm 4) on a panel of 519 sera in a regional reference laboratory in Lubumbashi, Zaire. Combining the Vironostika anti-HTLV-III ELISA with HIV Chek 1 + 2 or Clonatec Rapid HIV 1/2 Ab on all samples and retesting the discrepant results in WB or a line immunoassay (INNO-LIA) (algorithm 4), yielded a sensitivity of 100% and specificities of 98.4% and 99.0% respectively, at costs of 7.3 US $ and 9.3 US $ per test, respectively, for a 40% prevalence of HIV antibody positive samples. The conventional algorithm scored a sensitivity of 97.1% and a specificity of 100% for 11.3 US $ per test. The testing strategy of combining HIV Chek 1 + 2 and Clonatec Rapid HIV 1/2 Ab, an interesting option for small isolated centra, had a 96.6% sensitivity, but yielded only a slightly better specificity of 99.0%, as compared to 97.8% for HIV Chek alone. The price of combining the two simple assays using algorithm 3 was 6.8 US $ per test, using algorithm 4 was 10.6 US $. HIV testing strategies based on ELISA and a simple HIV test are a valuable alternative for reference laboratories faced with a high prevalence of HIV positive samples. PMID:1789703

  4. A New Reporter Cell Line to Monitor HIV Infection and Drug Susceptibility in vitro

    NASA Astrophysics Data System (ADS)

    Gervaix, Alain; West, Daniel; Leoni, Lorenzo M.; Richman, Douglas D.; Wong-Staal, Flossie; Corbeil, Jacques

    1997-04-01

    Determination of HIV infectivity in vitro and its inhibition by antiretroviral drugs by monitoring reduction of production of p24 antigen is expensive and time consuming. Such assays also do not allow accurate quantitation of the number of infected cells over time. To develop a simple, rapid, and direct method for monitoring HIV infection, we generated a stable T-cell line (CEM) containing a plasmid encoding the green fluorescent protein (humanized S65T GFP) driven by the HIV-1 long terminal repeat. Clones were selected that displayed low constitutive background fluorescnece, but a high level of GFP expression upon infection with HIV. HIV-1 infection induced a 100- to 1,000-fold increase in relative fluorescence of cells over 2 to 4 days as monitored by fluorescence microscopy, cytofluorimetry, and flow cytometry. Addition of inhibitors of reverse transcriptase, protease, and other targets at different multiplicities of infection permitted the accurate determination of drug susceptibility. This technique also permitted quantitation of infectivity of viral preparations by assessment of number of cells infected in the first round of infection. In conclusion, the CEM-GFP reporter cell line provides a simple, rapid, and direct method for monitoring HIV infectivity titers and antiretroviral drug susceptibility of syncytium-inducing strains.

  5. Preclinical and Clinical Performance of the Efoora Test, a Rapid Test for Detection of Human Immunodeficiency Virus-Specific Antibodies

    PubMed Central

    Arens, Max Q.; Mundy, Linda M.; Amsterdam, Daniel; Barrett, J. Tom; Bigg, Dan; Bruckner, David; Hanna, Bruce; Prince, Harry; Purington, Timothy; Hanna, Todd; Hewitt, Ross; Kalinka, Carolyn; Koppes, Thomas; Maxwell, Sarz; Moe, Ardis; Doymaz, Mehmet; Poulter, Melinda; Saber-Tehrani, Maryam; Simard, Lorenzo; Wilkins-Carmody, Donna; Vidaver, John; Berger, Cheryl; Davis, Alan H.; Alzona, Mortimer T.

    2005-01-01

    Barriers to effective diagnostic testing for human immunodeficiency virus type 1 (HIV-1) infection can be reduced with simple, reliable, and rapid detection methods. Our objective was to determine the accuracy, sensitivity, and specificity of a new rapid, lateral-flow immunochromatographic HIV-1 antibody detection device. Preclinical studies were performed using seroconversion, cross-reaction, and interference panels, archived clinical specimens, and fresh whole blood. In a multicenter, prospective clinical trial, a four-sample matrix of capillary (fingerstick) whole-blood specimens and venous whole blood, plasma, and serum was tested for HIV-1 antibodies with the Efoora HIV rapid test (Efoora Inc., Buffalo Grove, IL) and compared with an enzyme immunoassay (EIA) (Abbott Laboratories) licensed by the Food and Drug Administration. Western blot and nucleic acid test supplemental assays were employed to adjudicate discordant samples. Preclinical testing of seroconversion panels showed that antibodies were often detected earlier by the rapid test than by a reference EIA. No significant interference or cross-reactions were observed. Testing of 4,984 archived specimens yielded a sensitivity of 99.2% and a specificity of 99.7%. A prospective multicenter clinical study with 2,954 adult volunteers demonstrated sensitivity and specificity for the Efoora HIV rapid test of 99.8% (95% confidence interval [CI], 99.3 and 99.98%) and 99.0% (95% CI, 98.5 and 99.4%), respectively. Reactive rapid HIV-1 antibody detection was confirmed in 99.6% of those with a known HIV infection (n = 939), 5.2% of those in the high-risk group (n = 1,003), and 0.1% of those in the low-risk group (n = 1,012). For 21 (0.71%) patients, there was discordance between the results of the rapid test and the confirmatory EIA/Western blot tests. We conclude that the Efoora HIV rapid test is a simple, rapid assay for detection of HIV-1 antibodies, with high sensitivity and specificity compared to a standardized

  6. Simple, Inexpensive, and Rapid Approach to Fabricate Cross-Shaped Memristors Using an Inorganic-Nanowire-Digital-Alignment Technique and a One-Step Reduction Process.

    PubMed

    Xu, Wentao; Lee, Yeongjun; Min, Sung-Yong; Park, Cheolmin; Lee, Tae-Woo

    2016-01-20

    A rapid, scalable, and designable approach to produce a cross-shaped memristor array is demonstrated using an inorganic-nanowire digital-alignment technique and a one-step reduction process. Two-dimensional arrays of perpendicularly aligned, individually conductive Cu-nanowires with a nanometer-scale Cux O layer sandwiched at each cross point are produced. PMID:26585580

  7. Development of a quick and simple detection methodology for foot-and-mouth disease virus serotypes O, A and Asia 1 using a generic RapidAssay Device

    PubMed Central

    2013-01-01

    Background Outbreaks of Foot-and-mouth disease (FMD) have resulted in tremendous economic losses. Thus, the development of a rapid and easily performed test for FMD detection is important for controlling a FMD outbreak and containing its spread. The purpose of this project is to develop a lateral flow immunochromatographic (LFI) strip test for rapid detection of FMD virus serotypes O, A and Asia 1. Methods Specific monoclonal antibodies (mAbs) against each serotype were produced and used as the capture mAbs. A serotype independent mAb was selected and used as the detection mAb with the aim of subsequently developing a multi-serotype strip test. A new generation of the generic RapidAssay Device (gRAD) was used for the test. Result Each strip test can specifically detect the FMDV O, A or Asia 1 viruses, but not other vesicular disease viruses. The LFI strip tests for serotypes A and Asia 1 were able to identify all tested serotype A (n= 39) and Asia 1 field isolates (n=17). Whereas the test for serotype O detected 45 out of 46 field isolates. The sensitivity of this strip test was comparable with the double antibody sandwich ELISA for viral antigen detection. All vesicular fluid and epithelium samples collected from experimentally infected animals with serotype O, A and Asia 1 were identified as positive by the LFI strip test. Swab samples (n=11) collected over the lesion area from experimentally inoculated animals (serotype A) were examined. All of them demonstrated positive results using the LFI serotype A strip test and double antibody sandwich (DAS) ELISA. Conclusions The ability of strip tests to produce rapid results and high specificity makes it a valuable tool for early detection of FMDV O, A and Asia 1 in the field. PMID:23607273

  8. Development of simple and rapid elution methods for proteins from various affinity beads for their direct MALDI-TOF downstream application.

    PubMed

    Mlynarcik, Patrik; Bencurova, Elena; Madar, Marian; Mucha, Rastislav; Pulzova, Lucia; Hresko, Stanislav; Bhide, Mangesh

    2012-07-19

    Commercially available desalting techniques, necessary for downstream MALDI-TOF analysis of proteins, are often costly or time consuming for large-scale analysis. Here, we present techniques to elute proteins from various affinity resins, free from salt and ready for MALDI mass spectrometry. We showed that 0.1% TFA in 50% acetonitrile or 40% ethanol can be used as salt-free eluents for His-tagged proteins from variety of polyhistidine-affinity resins, while washing of resin beads twice with double-distilled water prior to the elution effectively desalted and recovered wide-range-molecular size proteins than commercially available desalting devices. Modified desalting and elution techniques were also applied for Flag- and Myc-tag affinity resins. The technique was further applied in co-precipitation assay, where the maximum recovery of wide-range molecular size proteins is crucial. Further, results showed that simple washing of the beads with double distilled water followed by elution with acetonitrile effectively desalted and recovered 150 kDa factor H protein of the sheep and its binding partner ~30 kDa BbCRASP-1 in co-precipitation assay. In summary, simple modifications in the desalting and elution strategy save time, labor and cost of the protein preparation for MALDI mass spectrometry; and large-scale protein purifications or co-precipitations can be performed with ease. PMID:22433248

  9. A simple Cr(VI)-S(IV)-O2 system for rapid and simultaneous reduction of Cr(VI) and oxidative degradation of organic pollutants.

    PubMed

    Yuan, Yanan; Yang, Shaojie; Zhou, Danna; Wu, Feng

    2016-04-15

    Hexavalent chromium (Cr(VI)), a heavy-metal contaminant, can be easily reduced to less toxic trivalent chromium (Cr(III)) by sulfite ions (S(IV)). However, S(IV) has not drawn as much attention as the ferrous ion has. We report herein a novel Cr(VI)-S(IV)-O2 system containing sulfite ions that rapidly and simultaneously reduces Cr(VI) and oxidize organic pollutants in the presence of oxygen in aqueous solutions. This Cr(VI)-S(IV)-O2 system contains the initiator Cr(VI), the reductant S(IV), and the oxidant O2, which produce oxysulfur radicals (mainly SO4(-) and SO5(-)) and hydroxyl radicals (OH). The Cr(VI)/S(IV) molar ratio, pH, and oxygen content play important roles in the entire reaction system. Acidic conditions (pH 3.0) facilitated degradation of organic compounds and reduction of Cr(VI) as well. In addition, experiments of rapid degradation of several kinds of organic pollutants such as azo dye (acid orange 7, AO7), aniline, phenol, bisphenol A etc were also conducted. Preliminary results show that the removal rates of the analogs of phenols or aromatic amines in this Cr(VI)-S(IV)-O2 system have a relationship with the electronic parameters (Hammett constant, σ) of the substituted groups. Thus, the Cr(VI)-S(IV)-O2 system, provides an excellent strategy of "waste control by waste" for removing multiple industrial contaminants. PMID:26799220

  10. Rapid formation of a superhydrophobic surface on a magnesium alloy coated with a cerium oxide film by a simple immersion process at room temperature and its chemical stability.

    PubMed

    Ishizaki, Takahiro; Saito, Naobumi

    2010-06-15

    We have developed a facile, simple, time-saving method of creating a superhydrophobic surface on a magnesium alloy by a simple immersion process at room temperature. First, a crystalline CeO(2) film was vertically formed on the magnesium alloy by immersion in a cerium nitrate aqueous solution for 20 min. The density of the crystals vertically with respect to the magnesium alloy increased with increasing immersion time. Next, the film were covered with fluoroalkylsilane (FAS: CF(3)(CF(2))(7)CH(2)CH(2)Si(OCH(3))(3)) molecules within 30 min by immersion in a toluene solution containing FAS and tetrakis(trimethylsiloxy)titanium (TTST: (CH(3))(3)SiO)(4)Ti). TTST was used as a catalyst to promote the hydrolysis and/or polymerization of FAS molecules. The FAS-coated CeO(2) film had a static contact angle of more than 150 degrees, that is, a superhydrophobic property. The shortest processing time for the fabrication of the superhydrophobic surface was 40 min. The contact angle hysteresis decreased with an increase in the immersion time in the cerium nitrate aqueous solution. The chemical stability of the superhydrophobic surface on magnesium alloy AZ31 was investigated. The average static water contact angles of the superhydrophobic surfaces after immersion in the solutions at pH 4, 7, and 10 for 24 h were found to be 139.7 +/- 2, 140.0 +/- 2, and 145.7 +/- 2 degrees, respectively. In addition, the chemical stability of the superhydrophobic surface in the solutions at pH ranging from 1 to 14 was also examined. The superhydrophobic surfaces had static contact angles of more than 142 degrees in the solutions at pH ranging from 1 to 14, showing that our superhydrophobic surface had a high chemical stability. Moreover, the corrosion resistance of the superhydrophobic surface on the magnesium alloy was investigated using electrochemical measurements. PMID:20377219

  11. RADMAP: Simple probes for rapid assessment of complex reactivity: A method and case studies on the reaction of hydrogen atoms with unsaturated organic molecules.

    PubMed

    Long, Andrew K; Fawcett, Jason A; Clyburne, Jason A C; Pye, Cory C

    2016-03-01

    RADMAP, an open source program, allows for rapid analysis and visualization of the earliest stages of reactions between any molecule and a monoatomic probe (i.e., H*, H(+), H(-), Br*, or any other monoatomic species) using ab initio methods. This program creates non-planar potential energy surfaces of the initial interaction between a molecule of interest and the monoatomic probe. These surfaces can be used to both predict the site of addition as well as provide a qualitative estimate for the relative proportion of the formation of adducts; therefore, it gives insight into both the reactivity and the kinetic stability of a molecule. The program presents a way to quickly predict the number of signals anticipated in transverse field muon spin resonance spectra as well as their relative intensities. PMID:26851865

  12. Simple and rapid method for the determination of the diastereomers of difenacoum in blood and liver using high-performance liquid chromatography with fluorescence detection.

    PubMed

    Kelly, M J; Chambers, J; MacNicoll, A D

    1993-10-22

    A rapid and sensitive high-performance liquid chromatographic method for the analysis of cis and trans diastereomers of the anticoagulant rodenticide difenacoum has been described. The methodology demonstrates potential for the analysis of diastereomers of related 4-hydroxycoumarin anticoagulants. Separations were achieved by reversed-phase chromatography on a Zorbax ODS column with gradient elution using acetonitrile-water, modified with 0.1% acetic acid, as the mobile phase. Detection of the analytes was effected by fluorescence at excitation and emission wavelengths of 310 and 390 nm, respectively. Sample preparation from both plasma and liver has been simplified to reduce preparation time and manipulation. The minimum detectable concentration of each diastereomer was 5 ng/ml. Recoveries of 100% were obtained from plasma and 93% from liver tissue. This method has been used for the investigation of the pharmacokinetics of difenacoum diastereomers in rats, and for investigation of unexplained hypoprothrombinaemic events encountered clinically. PMID:8106576

  13. A Simple Modification to the Mosquito Homogenization Protocol Safely Inactivates West Nile Virus and Allows Virus Detection by the Rapid Analyte Measurement Platform (RAMP®) ASSAY.

    PubMed

    Burkhalter, Kristen L; Biggerstaff, Brad J; Horiuchi, Kalanthe; Savage, Harry M

    2016-06-01

    We evaluated the ability of the Rapid Analyte Measurement Platform (RAMP(®)) mosquito-grinding buffer to inactivate West Nile virus (WNV) by subjecting WNV-positive samples ground in RAMP buffer to incubation intervals ranging from 5 min to 60 min. At each time point an aliquot was removed and serially diluted in bovine albumin (BA)-1 cell culture media to stop the inactivation process by RAMP buffer. Each BA-1 sample was tested for viable virus using Vero 6-well cell culture plaque assay and observed for plaques. We observed very limited inactivation of WNV (1-2 log10 plaque-forming units/ml) by RAMP buffer. Concerned for RAMP operators who may be using this assay in low-level biocontainment facilities, we developed an alternate sample homogenization protocol using Triton X-100 detergent that ensures complete WNV inactivation without compromising the performance of the RAMP assay. PMID:27280345

  14. A simple method for the rapid determination of the stereospecificity of NAD-dependent dehydrogenases applied to mammalian IMP dehydrogenase and bacterial NADH peroxidase.

    PubMed

    Cooney, D; Hamel, E; Cohen, M; Kang, G J; Dalal, M; Marquez, V

    1987-11-01

    The stereospecificity of IMP dehydrogenase (IMP:NAD+ oxidoreductase, EC 1.1.1.205) from two different sources was determined. The enzyme preparations were obtained from murine lymphoblasts and from Escherichia coli. Both enzymes transferred the 2-3H of IMP to the pro-S position of carbon atom C-4 of the nicotinamide ring in NAD. Thus, B-sided stereospecificity is common to the enzyme from two very different species. In addition, the studies described here demonstrate that alcohol dehydrogenase and NADH peroxidase, used as auxiliary enzymes, in combination with a microdistillation procedure, should permit rapid determination of the stereospecificity of any NAD-dependent dehydrogenase for which the appropriate tritiated substrate is available. PMID:2889473

  15. A rapid and simple pipeline for synthesis of mRNA-ribosome-V(H)H complexes used in single-domain antibody ribosome display.

    PubMed

    Bencurova, Elena; Pulzova, Lucia; Flachbartova, Zuzana; Bhide, Mangesh

    2015-06-01

    The single-domain antibody (VHH) is a promising building block for a number of antibody-based applications. Ribosome display can successfully be used in the production of VHH. However, the construction of the expression cassette, confirmation of the translation and proper folding of the nascent chain, and the purification of the ribosome complexes, remain cumbersome tasks. Additionally, selection of the most suitable expression system can be challenging. We have designed primers that will amplify virtually all Camelidae VHH. With the help of a double-overlap extension (OE) polymerase chain reaction (PCR) we have fused VHH with the F1 fragment (T7 promoter and species-independent translation sequence) and the F2 fragment (mCherry, Myc-tag, tether, SecM arrest sequence and 3' stem loop) to generate a full-length DNA cassette. OE-PCR generated fragments were incubated directly with cell-free lysates (Leishmania torentolae, rabbit reticulocyte or E. coli) for the synthesis of mRNA-VHH-mCherry-ribosome complexes in vitro. Alternatively, the cassette was ligated in pQE-30 vector and transformed into E. coli to produce ribosome complexes in vivo. The results showed that the same expression cassette could be used to synthesize ribosome complexes with different expression systems. mCherry reporter served to confirm the synthesis and proper folding of the nascent chain, Myc-tag was useful in the rapid purification of ribosome complexes, and combination of the SecM sequence and 3' stem loop made the cassette universal, both for cells-free and E. coli in vivo. This rapid and universal pipeline can effectively be used in antibody ribosome display and VHH production. PMID:25902394

  16. Modification and evaluation of the Carba NP test by use of paper strip for simple and rapid detection of carbapenemase-producing Enterobacteriaceae.

    PubMed

    Srisrattakarn, Arpasiri; Lulitanond, Aroonlug; Wilailuckana, Chotechana; Charoensri, Nicha; Wonglakorn, Lumyai; Piyapatthanakul, Sirikan; Supajeen, Ampai; Chanawong, Aroonwadee

    2016-07-01

    Carbapenemase-producing Enterobacteriaceae (CPE) isolates have now emerged worldwide. We therefore modified the phenotypic Carba NP test by use of a filter paper strip for easily and rapidly identifying CPE in routine laboratory. A collection of 56 CPE and carbapenemase-producing Pseudomonas spp. isolates (including 28 NDM-1, 11 IMP-14a, 1 IMP-1, 1 IMP-4, 1 IMP-9, 1 IMP-15, 4 VIM-2, 1 VIM-1, 1 IMP-14a with VIM-2, 3 OXA-48, 3 OXA-181 and 1 KPC-2 producers) and 41 non-CPE isolates (including 19 ESBL, 7 pAmpC, 3 AmpC, 9 ESBL with pAmpC and 3 non-ESBL & non-AmpC producers) as confirmed by the PCR methods were tested by the paper strip method using pharmaceutical imipenem/cilastatin as a substrate. Bacterial colonies of each isolate were applied directly on filter paper strips dropped with either imipenem-phenol red (test strip) or phenol red solution alone (control strip). The reaction was read within 5 min. This test failed to detect 3 OXA-181, 2 OXA-48 and 3 IMP-14a producers (85.7 % sensitivity), whereas no false positives were seen (100 % specificity). Further evaluation of the paper strip test in 267 CPE screening-positive isolates from three hospitals by their medical technologists showed 92.0 % sensitivity (100 % for NDM producers) and 100 % specificity compared with the PCR methods. Because of its ease, rapidness and cost effective, the paper strip test has a potential for routine CPE testing in low-resource laboratories particularly in areas with high prevalence of NDM enzymes, leading to appropriate antimicrobial therapy and infection control strategy. PMID:27263012

  17. HIV Symptoms

    MedlinePlus

    ... Submit Home > HIV/AIDS > What is HIV/AIDS? HIV/AIDS This information in Spanish ( en español ) HIV symptoms Photo courtesy of AIDS.gov More information ... and brain Return to top More information on HIV symptoms Explore other publications and websites Basic Information ...

  18. A microfluidic device for practical label-free CD4(+) T cell counting of HIV-infected subjects.

    PubMed

    Cheng, Xuanhong; Irimia, Daniel; Dixon, Meredith; Sekine, Kazuhiko; Demirci, Utkan; Zamir, Lee; Tompkins, Ronald G; Rodriguez, William; Toner, Mehmet

    2007-02-01

    Practical HIV diagnostics are urgently needed in resource-limited settings. While HIV infection can be diagnosed using simple, rapid, lateral flow immunoassays, HIV disease staging and treatment monitoring require accurate counting of a particular white blood cell subset, the CD4(+) T lymphocyte. To address the limitations of current expensive, technically demanding and/or time-consuming approaches, we have developed a simple CD4 counting microfluidic device. This device uses cell affinity chromatography operated under differential shear flow to specifically isolate CD4(+) T lymphocytes with high efficiency directly from 10 microliters of unprocessed, unlabeled whole blood. CD4 counts are obtained under an optical microscope in a rapid, simple and label-free fashion. CD4 counts determined in our device matched measurements by conventional flow cytometry among HIV-positive subjects over a wide range of absolute CD4 counts (R(2) = 0.93). This CD4 counting microdevice can be used for simple, rapid and affordable CD4 counting in point-of-care and resource-limited settings. PMID:17268618

  19. Simple combination of oxidants with zero-valent-iron (ZVI) achieved very rapid and highly efficient removal of heavy metals from water.

    PubMed

    Guo, Xuejun; Yang, Zhe; Dong, Haiyang; Guan, Xiaohong; Ren, Qidong; Lv, Xiaofang; Jin, Xin

    2016-01-01

    This study, for the first time, demonstrated a continuously accelerated Fe(0) corrosion driven by common oxidants (i.e., NaClO, KMnO4 or H2O2) and thereby the rapid and efficient removal of heavy metals (HMs) by zero-valent iron (ZVI) under the experimental conditions of jar tests and column running. ZVI simply coupled with NaClO, KMnO4 or H2O2 (0.5 mM) resulted in almost complete As(V) removal within only 10 min with 1000 μg/L of initial As(V) at initial pH of 7.5(±0.1) and liquid solid ratio of 200:1. Simultaneous removal of 200 μg/L of initial Cd(II) and Hg(II) to 2.4-4.4 μg/L for Cd(II) and to 4.0-5.0 μg/L for Hg(II) were achieved within 30 min. No deterioration of HM removal was observed during the ten recycles of jar tests. The ZVI columns activated by 0.1 mM of oxidants had stably treated 40,200 (NaClO), 20,295 (KMnO4) and 40,200 (H2O2) bed volumes (BV) of HM-contaminated drinking water, but with no any indication of As breakthrough (<10 μg/L) even at short empty bed contact time (EBCT) of 8.0 min. The high efficiency of HMs removal from both the jar tests and column running implied a continuous and stable activation (overcoming of iron passivation) of Fe(0) surface by the oxidants. Via the proper increase in oxidant dosing, the ZVI/oxidant combination was applicable to treat highly As(V)-contaminated wastewater. During Fe(0) surface corrosion accelerated by oxidants, a large amount of fresh and reactive iron oxides and oxyhydroxides were continuously generated, which were responsible for the rapid and efficient removal of HMs through multiple mechanisms including adsorption and co-precipitation. A steady state of Fe(0) surface activation and HM removal enabled this simply coupled system to remove HMs with high speed, efficiency and perdurability. PMID:26575476

  20. Development of a simple, rapid, and robust liquid chromatographic method for the simultaneous determination of sulfalene, sulfadoxine, and pyrimethamine in tablets.

    PubMed

    Mwalwisi, Yonah H; Hoellein, Ludwig; Kaale, Eliangiringa; Holzgrabe, Ulrike

    2016-09-10

    A simple, cost effective, accurate, and precise RP-HPLC method was developed for the simultaneous determination of sulfalene and sulfadoxine in fixed dose dual combinations with pyrimethamine together with their related substances. Proprietary products containing these combinations are often being prescribed in malaria endemic countries. Quantification of the active compounds and impurity profiling was achieved using two standard C18 columns with a mobile phase being composed of 60% (v/v) of a 0.05M KH2PO4 buffer solution (pH=2.6) and 40% (v/v) of methanol, applying an isocratic elution mode and a detection wavelength of 215nm. The method allows a quick quantitative determination of sulfadoxine and sulfalene and the separation of the respective impurities within a total runtime of approximately 15min and was validated with respect to specificity, linearity, precision, accuracy, limits of detection and quantification, robustness, and stability of the standard and sample solutions. The method is simpler than the corresponding method described in the International Pharmacopoeia and the United States Pharmacopoeia in terms of being easy to apply, being less time consuming, and utilizing reagents and chemicals which are cost efficient. PMID:27505128

  1. A simple and rapid method to identify and quantitatively analyze triterpenoid saponins in Ardisia crenata using ultrafast liquid chromatography coupled with electrospray ionization quadrupole mass spectrometry.

    PubMed

    Ma, Ling; Li, Wei; Wang, Hanqing; Kuang, Xinzhu; Li, Qin; Wang, Yinghua; Xie, Peng; Koike, Kazuo

    2015-01-01

    Ardisia plant species have been used in traditional medicines, and their bioactive constituents of 13,28-epoxy triterpenoid saponins have excellent biological activities for new drug development. In this study, a fast and simple method based on ultrafast liquid chromatography coupled to electrospray ionization mass spectrometry (UFLC-MS) was developed to simultaneously identify and quantitatively analyze triterpenoid saponins in Ardisia crenata extracts. In total, 22 triterpenoid saponins, including two new compounds, were identified from A. crenata. The method exhibited good linearity, precision and recovery for the quantitative analysis of eight marker saponins. A relative quantitative method was also developed using one major saponin (ardisiacrispin B) as the standard to break through the choke-point of the lack of standards in phytochemical analysis. The method was successfully applied to quantitatively analyze saponins in commercially available plant samples. This study describes the first systematic analysis of 13,28-epoxy-oleanane-type triterpenoid saponins in the genus Ardisia using LC-ESI-MS. The results can provide the chemical support for further biological studies, phytochemotaxonomical studies and quality control of triterpenoid saponins in medicinal plants of the genus Ardisia. PMID:25459939

  2. Simple, sensitive and rapid LC-MS method for the quantitation of indapamide in human plasma--application to pharmacokinetic studies.

    PubMed

    Chen, Wei-Dong; Liang, Yan; Zhang, Hong; Li, Hao; Xiong, Ye; Wang, Guang-Ji; Xie, Lin

    2006-09-14

    A sensitive and specific method using liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS) has been developed and validated for the identification and quantification of indapamide in human plasma. A simple liquid-liquid extraction procedure was followed by injection of the extracts on to a C18 column with gradient elution and detection using a single quadrupole mass spectrometer in selected ion monitoring (SIM) mode. The method was tested using six different plasma batches. Linearity was established for the concentration range 0.5-100.0 ng/ml, with a coefficient of determination (r) of 0.9998 and good back-calculated accuracy and precision. The intra- and inter-day precision (RSD%) was lower than 10%, and accuracy ranged from 85 to 115%. The lower limit of quantification was reproducible at 0.2 ng/ml with 0.2 ml plasma. The proposed method enables the unambiguous identification and quantification of indapamide for pre-clinical and clinical studies. PMID:16843739

  3. A simple and rapid method for the multielement analysis of wheat crispbread products by inductively coupled plasma-optical emission spectrometry.

    PubMed

    Szymczycha-Madeja, Anna

    2014-01-01

    A simple and fast method for the determination of total concentrations of Ba, Ca, Cd, Cr, Cu, Fe, Mg, Mn, Ni, P, Sr, and Zn in wheat crispbreads using inductively coupled plasma-optical emission spectrometry was developed. Initially, three different sample preparation procedures, i.e., microwave- assisted digestion in an HNO3 and H2O2 mixture and solubilization in aqua regia or tetramethylammonium hydroxide solution were compared. The performance of these procedures was evaluated for precision, accuracy, and LOD of the elements. It was established that the application of aqua regia allows determining elements with LOD within 0.1-42 ng/mL, precision of 0.2-4.1%, and accuracy better than 5%. Furthermore, good agreement between measured and certified values of a National Institute of Standards and Technology certified reference material of wheat flour (SRM 1567a) was found. It was confirmed that the proposed method could be used successfully as an alternative to microwave-assisted acid digestion in routine multielement analyses of wheat crispbreads. PMID:25632441

  4. Simple and rapid assay for effect of the new oral anticoagulant (NOAC) rivaroxaban: preliminary results support further tests with all NOACs

    PubMed Central

    2014-01-01

    Background New oral anticoagulant (NOAC) drugs are known to influence the results of some routine hemostasis tests. Further data are needed to enable routine assessment of the effects of NOAC on clotting parameters in some special circumstances. Methods Following administration of rivaroxaban to patients, at the likely peak and trough activity times, we assessed the effects on prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and clotting time using Russell’s viper venom, and in the presence of phospholipids and calcium reagent available as DVVreagent® and DVVconfirm®. The data were used to determine an adequate NOAC plasma level based on anticoagulant activities expressed as a ratio (patients/normal, R-C). Results DVVconfirm as R-C could be rapidly performed, and the results were reasonably sensitive for rivaroxaban and probably for other FX inhibitors. This assay is not influenced by lupus anticoagulant and heparin, does not require purified NOAC as control, and will measure whole-plasma clotting activity. Conclusions We propose a cut-off R-C value of 4.52 ± 0.33 for safety, but clinical studies are needed to establish whether this cut-off is useful for identifying patients at increased risk of hemorrhage or exhibiting low anticoagulation effect. It also seems possible that normal R-C could indicate an absence of anticoagulant activity when rivaroxaban is discontinued due to episodes of uncontrolled bleeding during anticoagulation or for emergency surgery. PMID:24656069

  5. A simple and rapid flow-injection chemiluminescence method for the determination of noscapine with Ru(phen)3(2+)-Ce(IV) system.

    PubMed

    Rezaei, Behzad; Mokhtari, Ali; Khayamian, Taghi

    2007-08-01

    A new flow injection chemiluminescence (CL) system was used for the determination of noscapine. This technique is based on the reduction effect of noscapine on the Ru(phen)3(3+), which is produced by reaction between Ru(phen)3(2+) and acidic Ce(IV) solutions, and this rapid reduction produces strong CL. Calibration plots were linear over the range of 3.0 x 10(-7) - 2.0 x 10(-6) mol L(-1) and 2.0 x 10(-6) - 2.0 x 10(-4) mol L(-1). The CL intensity was so high, that it is able to produce a detection limit of 6.6 x 10(-8) M noscapine (3sigma). The relative standard deviation of 2.0 x 10(-6) M noscapine was 1.0% (n=10). The proposed method was successfully applied for the flow injection determination of noscapine in cough and Tonin syrup samples. The results of real sample analyses show good recovery percentages (97.3-102.4%). The minimum sampling rate was 100 samples per hour. PMID:17899875

  6. Development of a novel real-time PCR-based strategy for simple and rapid molecular pathotyping of Newcastle disease virus.

    PubMed

    Yacoub, Alia; Leijon, Mikael; McMenamy, Michael J; Ullman, Karin; McKillen, John; Allan, Gordon; Belák, Sándor

    2012-05-01

    A novel real-time PCR strategy was applied to simultaneously detect and to discriminate low-pathogenic lentogenic and virulent meso/velogenic Newcastle disease virus (NDV). The pathotyping is achieved by a three-step semi-nested PCR. A pre-amplification of the cleavage site (CS) region of the F gene is followed by a two-level duplex real-time PCR directly targeting the CS, combining detection and pathotyping in a single tube. A wide range of NDV isolates spanning all genotypes were successfully detected and pathotyped. Clinical samples from outbreaks in Sweden in 2010 that were positive by the novel PCR method were also successfully pathotyped. The method is time-saving, reduces labour and costs and provides opportunities for rapid diagnosis at remote locations and in the field. Since the same strategy was also recently applied to avian influenza virus pathotyping, it shows promise of finding broad utility in diagnostics of infectious diseases caused by different RNA viruses in various hosts. PMID:22302287

  7. HIV Prevention

    MedlinePlus

    ... to treat HIV infection (called antiretroviral therapy, or ART) the right way, every day and his or ... way, every day, the medicine to treat HIV (ART) reduces the amount of HIV (called “viral ...

  8. AAV2/8 Vectors Purified from Culture Medium with a Simple and Rapid Protocol Transduce Murine Liver, Muscle, and Retina Efficiently

    PubMed Central

    Doria, Monica; Ferrara, Antonella

    2013-01-01

    Abstract During the production of some adeno-associated virus (AAV) serotypes, a large amount of vectors is found in the medium of producing cells. For their purification, previous protocols used tangential flow filtration (TFF) of the medium followed by iodixanol gradient centrifugation. Taking advantage of the higher purity of the medium than the cell-derived material as the source of AAV, we tested a simple method that combines production of large culture medium volumes containing AAV from cell stacks with medium clarification+TFF without further time-consuming and nonscalable centrifugation. To test this, we selected AAV2/8, which is emerging as a favored serotype for transduction of liver, muscle, and retina and abundantly found in the extracellular medium. We show that yields and in vitro infectivity of AAV2/8 vectors produced from the culture medium using this method are higher than those of vectors purified from the same cell lysate using a conventional CsCl2 gradient ultracentrifugation-based method, although purity appears inferior. In addition, we found that the transduction efficiency of AAV2/8 purified from medium was similar to that of AAV2/8 purified from the same cell lysate in the murine liver, muscle, and retina. Considering that the purification protocol from the medium we describe requires 3 hr as opposed to the 63 hr of a conventional two-round CsCl2-gradient ultracentrifugation+desalting, we conclude that TFF of the medium containing AAV2/8 represents a quick and scalable method to purify research-grade vectors for use in animal models. PMID:24116943

  9. Predicting rapid herbicide leaching to surface waters from an artificially drained headwater catchment using a one dimensional two-domain model coupled with a simple groundwater model.

    PubMed

    Tediosi, A; Whelan, M J; Rushton, K R; Gandolfi, C

    2013-02-01

    Pesticide losses to water can present problems for environmental management, particularly in catchments where surface waters are abstracted for drinking water supply. The relative role of different transfer pathways (spray drift, spills, overland flow and leaching from soils) is often uncertain, and there is a need for experimental observation and modelling to ensure that processes are understood under a range of conditions. Here we examine the transport of propyzamide and carbetamide in a small (15.5 ha) headwater sub-catchment dominated by an artificially drained field with strongly undulating topography (topographic gradients >1:10). Specifically, we explore the validity of the "field-scale lysimeter" analogy by applying the one dimensional mathematical model MACRO. Although one dimensional representation has been shown to be reasonable elsewhere, the scale and topography of the monitored system challenge many of the underlying assumptions. MACRO considers two interacting flow domains: micropores and macropores. The effect of subsurface drains can also be included. A component of the outflow from the main drain was identified as originating from an upslope permeable shallow aquifer which was represented using a simple groundwater model. Predicted herbicide losses were sensitive to drain spacing and the organic carbon to water partition coefficient, K(OC). The magnitude of the peak water and herbicide transport and their timing were simulated satisfactorily, although model performance was poor following a period of one month when snow covered the ground and precipitation was underestimated by the rain gauge. Total herbicide loads were simulated adequately by MACRO, suggesting that the field-scale lysimeter analogy is valid at this scale, although baseflow contributions to flow needed to be accounted for separately in order to adequately represent hydrological response. PMID:23313906

  10. Predicting rapid herbicide leaching to surface waters from an artificially drained headwater catchment using a one dimensional two-domain model coupled with a simple groundwater model

    NASA Astrophysics Data System (ADS)

    Tediosi, A.; Whelan, M. J.; Rushton, K. R.; Gandolfi, C.

    2013-02-01

    Pesticide losses to water can present problems for environmental management, particularly in catchments where surface waters are abstracted for drinking water supply. The relative role of different transfer pathways (spray drift, spills, overland flow and leaching from soils) is often uncertain, and there is a need for experimental observation and modelling to ensure that processes are understood under a range of conditions. Here we examine the transport of propyzamide and carbetamide in a small (15.5 ha) headwater sub-catchment dominated by an artificially drained field with strongly undulating topography (topographic gradients > 1:10). Specifically, we explore the validity of the "field-scale lysimeter" analogy by applying the one dimensional mathematical model MACRO. Although one dimensional representation has been shown to be reasonable elsewhere, the scale and topography of the monitored system challenge many of the underlying assumptions. MACRO considers two interacting flow domains: micropores and macropores. The effect of subsurface drains can also be included. A component of the outflow from the main drain was identified as originating from an upslope permeable shallow aquifer which was represented using a simple groundwater model. Predicted herbicide losses were sensitive to drain spacing and the organic carbon to water partition coefficient, KOC. The magnitude of the peak water and herbicide transport and their timing were simulated satisfactorily, although model performance was poor following a period of one month when snow covered the ground and precipitation was underestimated by the rain gauge. Total herbicide loads were simulated adequately by MACRO, suggesting that the field-scale lysimeter analogy is valid at this scale, although baseflow contributions to flow needed to be accounted for separately in order to adequately represent hydrological response.

  11. Inductively coupled plasma mass spectrometry as a simple, rapid, and inexpensive method for determination of uranium in urine and fresh water: Comparison with LIF

    SciTech Connect

    Karpas, Z.; Halicz, L.; Roiz, J.

    1996-12-01

    A simple method, based on inductively coupled plasma mass spectrometry, for determination of uranium in urine at levels that indicate occupational exposure, is presented. Sample preparation involves a fifty-fold dilution of the urine by nitric acid (2% HNO{sub 3}) and no other chemical treatment or separation. The analysis itself is completed in under 3 min. The analytical procedure is fully autominated so that a technician may perform over 100 analyses per day. With proper control of the blank contribution, a lower limit of detection of 3 ng L{sup {minus}1} in the original urine sample was achieved. Uranium concentrations in the range 6-30 ng L{sup {minus}1} were found in urine samples of people that are not occupationally exposed. The validity of the results was demonstrated through measurement of standards, controlled uranium addition experiments and, at higher concentrations, by comparison with results obtained by an independent method based on laser induced fluorescence. The laser induced fluorescence technique was found to be sufficient for detection of occupational exposure at an action level of 1.5 {mu}g L{sup {minus}1}. Use of internal standards, indium, and thallium, improved quantification by about 10%, but was not deemed necessary for routine analysis. The inductively coupled plasma mass spectrometry is also ideally suited for monitoring uranium in fresh water and drinking water, as no sample dilution is required and the lower limit of detection is below 0.15 ng L{sup {minus}1}. 41 refs., 4 figs., 5 tabs.

  12. Optimisation of the separation of four major neutral glycosphingolipids: application to a rapid and simple detection of urinary globotriaosylceramide in Fabry disease.

    PubMed

    Roy, S; Gaudin, K; Germain, D P; Baillet, A; Prognon, P; Chaminade, P

    2004-06-15

    A simple method for the separation of the four major neutral glycosphingolipids, present in all human tissue, was developed. This gradient normal phase-HPLC method utilises a polyvinyl alcohol bonded stationary phase and an evaporative light-scattering detection (ELSD). Screening pure solvents in a binary gradient elution mode allowed, in a first step, to assess the behaviour of the studied solutes and to select the solvents for further mobile phase optimisation. The proportion of the remaining solvents was defined to reach a maximal resolution. The reduction of the analysis time and the enhancement of the signal were obtained by optimising the gradient slope and the flow-rate. Optimal levels of triethylamine and formic acid (TEA-FA) for the enhancement of the evaporative light scattering detector response were established at 0.1% (v/v). Thus, the optimal conditions for the separation of the four glycosphingolipids was obtained with a gradient elution from a 100% chloroform to a 100% acetone:methanol (90:10 (v/v)) mobile phase at 0.2 ml min-1, using a 10% min-1 gradient slope. Finally, this method was applied to detect the excess of one of the neutral sphingolipids, namely globotriaosylceramide (Gb3) in the urine of patients affected with Fabry disease. A liquid-liquid extraction of the sediments obtained from an aliquot of only ten ml of urine proved sufficient to detect the excess of Gb3 present in both hemizygote and heterozygote patients. In all, the ability of our method to detect abnormal amounts of Gb3 in urinary sediments could allow the diagnosis of weakly symptomatic Fabry patients in large screening programs PMID:15135109

  13. Rapid and simple method for detecting the toxin B gene of Clostridium difficile in stool specimens by loop-mediated isothermal amplification.

    PubMed

    Kato, Haru; Yokoyama, Toshiyuki; Kato, Hideaki; Arakawa, Yoshichika

    2005-12-01

    We applied the loop-mediated isothermal amplification (LAMP) assay to the detection of the toxin B gene (tcdB) of Clostridium difficile for identification of toxin B (TcdB)-positive C. difficile strains and detection of tcdB in stool specimens. tcdB was detected in all toxin A (TcdA)-positive, TcdB-positive (A(+)B(+)) and TcdA-negative, TcdB-positive (A(-)B(+)) C. difficile strains but not from TcdA-negative, TcdB-negative strains. Of the 74 stool specimens examined, A(+)B(+) or A(-)B(+) C. difficile was recovered from 39 specimens, of which 38 specimens were LAMP positive and one was negative. Amplification was obtained in 10 specimens that were culture negative, indicating that LAMP is highly sensitive. The LAMP assay was applied to detection of tcdB in DNA extracted by a simple boiling method from 47 of those 74 specimens, which were cultured overnight in cooked-meat medium (CMM). Twenty-two of 24 culture-positive specimens were positive for LAMP on DNA from the culture in CMM. Four specimens were culture negative but positive by LAMP on DNA from CMM cultures. The LAMP assay is a reliable tool for identification of TcdB-positive C. difficile as well as for direct detection of tcdB in stool specimens with high sensitivity. Detection of tcdB by LAMP from overnight cultures in CMM could be an alternative method of diagnostic testing at clinical laboratories without special apparatus. PMID:16333105

  14. Simple and rapid fabrication of disposable carbon-based electrochemical cells using an electronic craft cutter for sensor and biosensor applications.

    PubMed

    Afonso, André S; Uliana, Carolina V; Martucci, Diego H; Faria, Ronaldo C

    2016-01-01

    This work describes the construction of an all-plastic disposable carbon-based electrochemical cell (DCell) using a simple procedure based on the use of a home cutter printer for prototyping and laminating. The cutter printer and adhesive vinyl films were used to produce three electrodes in an electrochemical cell layout, and a laminating process was then used to define the geometric area and insulate the electrodes. The DCell showed excellent performance in several applications including the determination of toxic metals in water samples, the immobilization of DNA and the detection of Salmonella. An unmodified DCell was applied for Pb and Cd detection in the range of 100-300 ng mL(-1) with a limit of detection of 50 and 39 ng mL(-1) for Cd and Pb, respectively. DNA was successfully immobilized on a DCell and used for studies of interaction between bisphenol A and DNA. The square wave voltammetry of a DNA modified DCell presented a guanine oxidation current 2.5 times greater after exposure of the electrode to bisphenol A and no current variation for the adenine moiety indicating that bisphenol A showed a preference for DNA interaction sites. A magneto-immunoassay was developed using a DCell for Salmonella detection in milk samples. The system presented a linear range from 100 to 700 cells mL(-1) with a limit of detection of 100 cells mL(-1) and good recovery values between 93% and 101% in milk samples, with no interference from Escherichia coli. Using the proposed method, hundreds of DCells can be assembled in less than two hours, at a material cost of less than US $0.02 per cell. The all-plastic disposable electrochemical cell developed was successfully applied as an electrochemical sensor and biosensor. The feasibility of the developed all-plastic disposable electrochemical cell was demonstrated in applications as both sensor and biosensor. PMID:26695279

  15. Rapid and Simultaneous Detection of Major Drug Resistance Mutations in Reverse Transcriptase Gene for HIV-1 CRF01_AE, CRF07_BC and Subtype B in China Using Sequenom MassARRAY® System.

    PubMed

    Cheung, Ka-Wai; Peng, Qiaoli; He, Liufen; Cai, Kanru; Jiang, Qiang; Zhou, Boping; To, Sabrina Wai-Chi; Yam, Wing-Cheong; Liu, Li; Chen, Zhiwei; Wang, Hui

    2016-01-01

    The development of a rapid, high-throughput and cost-effective HIV-1 drug resistance (HIV-DR) testing system is a challenge for areas consisting different HIV-1 strains. In this study, we established a broadly reactive multiplex assay that could simultaneously detect major drug resistance mutations at 8 loci, which are associated with resistance to commonly used nucleoside reverse transcriptase inhibitors (NRTIs) and Non-nucleoside reverse transcriptase inhibitors (NNRTIs), in specimens of HIV-1 CRF01_AE, CRF07_BC and subtype B, the three major circulating strains in China, using the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) provided by Sequenom MassARRAY® system. To establish the assay, pol gene fragments were prepared from the plasma viral RNA of 159 patients by nested PCR and the presence of wild type and mutant alleles at the 8 loci were analyzed by MALDI-TOF MS. In terms of loci, the detection rate of the alleles was greater than 97% for M41L, K65R, M184V and G190A, 91.2% for K101E/Q/P, 91.2% for T215F/Y, 89.9% for K103N/S and 80.5% for L210W. In terms of individuals, 80% of the alleles were detected in 95.4% CRF01_AE patients, 100% CRF07_BC patients and 83.3% subtype B patients. Importantly, the MALDI-TOF MS results were concordant to the drug resistance profiles of patients obtained from conventional sequencing analysis after excluded the failed detections. Using plasmid templates, the assay was estimated to be sensitive to detect drug resistant variants at level about 20% of the circulating viral population. The capability of this assay to detect mixed viral populations was further verified by two different patient specimens. In conclusion, this study evaluated the use of Sequenom MassARRAY® system for high-throughput detection of HIV-DR mutations towards the commonly used reverse transcriptase inhibitors in China. PMID:27092551

  16. Rapid and Simultaneous Detection of Major Drug Resistance Mutations in Reverse Transcriptase Gene for HIV-1 CRF01_AE, CRF07_BC and Subtype B in China Using Sequenom MassARRAY® System

    PubMed Central

    He, Liufen; Cai, Kanru; Jiang, Qiang; Zhou, Boping; To, Sabrina Wai-Chi; Yam, Wing-Cheong; Liu, Li; Chen, Zhiwei; Wang, Hui

    2016-01-01

    The development of a rapid, high-throughput and cost-effective HIV-1 drug resistance (HIV-DR) testing system is a challenge for areas consisting different HIV-1 strains. In this study, we established a broadly reactive multiplex assay that could simultaneously detect major drug resistance mutations at 8 loci, which are associated with resistance to commonly used nucleoside reverse transcriptase inhibitors (NRTIs) and Non-nucleoside reverse transcriptase inhibitors (NNRTIs), in specimens of HIV-1 CRF01_AE, CRF07_BC and subtype B, the three major circulating strains in China, using the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) provided by Sequenom MassARRAY® system. To establish the assay, pol gene fragments were prepared from the plasma viral RNA of 159 patients by nested PCR and the presence of wild type and mutant alleles at the 8 loci were analyzed by MALDI-TOF MS. In terms of loci, the detection rate of the alleles was greater than 97% for M41L, K65R, M184V and G190A, 91.2% for K101E/Q/P, 91.2% for T215F/Y, 89.9% for K103N/S and 80.5% for L210W. In terms of individuals, 80% of the alleles were detected in 95.4% CRF01_AE patients, 100% CRF07_BC patients and 83.3% subtype B patients. Importantly, the MALDI-TOF MS results were concordant to the drug resistance profiles of patients obtained from conventional sequencing analysis after excluded the failed detections. Using plasmid templates, the assay was estimated to be sensitive to detect drug resistant variants at level about 20% of the circulating viral population. The capability of this assay to detect mixed viral populations was further verified by two different patient specimens. In conclusion, this study evaluated the use of Sequenom MassARRAY® system for high-throughput detection of HIV-DR mutations towards the commonly used reverse transcriptase inhibitors in China. PMID:27092551

  17. Evaluation of two line probe assays for rapid detection of Mycobacterium tuberculosis, tuberculosis (TB) drug resistance, and non-TB Mycobacteria in HIV-infected individuals with suspected TB.

    PubMed

    Luetkemeyer, Anne F; Kendall, Michelle A; Wu, Xingye; Lourenço, Maria Cristina; Jentsch, Ute; Swindells, Susan; Qasba, Sarojini S; Sanchez, Jorge; Havlir, Diane V; Grinsztejn, Beatriz; Sanne, Ian M; Firnhaber, Cynthia

    2014-04-01

    Limited performance data from line probe assays (LPAs), nucleic acid tests used for the rapid diagnosis of tuberculosis (TB), nontuberculosis mycobacteria (NTM), and Mycobacterium tuberculosis drug resistance are available for HIV-infected individuals, in whom paucibacillary TB is common. In this study, the strategy of testing sputum with GenoType MTBDRplus (MTBDR-Plus) and GenoType Direct LPA (Direct LPA) was compared to a gold standard of one mycobacterial growth indicator tube (MGIT) liquid culture. HIV-positive (HIV(+)) individuals with suspected TB from southern Africa and South America with <7 days of TB treatment had 1 sputum specimen tested with Direct LPA, MTBDR-Plus LPA, smear microscopy, MGIT, biochemical identification of mycobacterial species, and culture-based drug-susceptibility testing (DST). Of 639 participants, 59.3% were MGIT M. tuberculosis culture positive, of which 276 (72.8%) were acid-fast bacillus (AFB) smear positive. MTBDR-Plus had a sensitivity of 81.0% and a specificity of 100%, with sensitivities of 44.1% in AFB smear-negative versus 94.6% in AFB smear-positive specimens. For specimens that were positive for M. tuberculosis by MTBDR-Plus, the sensitivity and specificity for rifampin resistance were 91.7% and 96.6%, respectively, and for isoniazid (INH) they were 70.6% and 99.1%. The Direct LPA had a sensitivity of 88.4% and a specificity of 94.6% for M. tuberculosis detection, with a sensitivity of 72.5% in smear-negative specimens. Ten of 639 MGIT cultures grew Mycobacterium avium complex or Mycobacterium kansasii, half of which were detected by Direct LPA. Both LPA assays performed well in specimens from HIV-infected individuals, including in AFB smear-negative specimens, with 72.5% sensitivity for M. tuberculosis identification with the Direct LPA and 44.1% sensitivity with MTBDR-Plus. LPAs have a continued role for use in settings where rapid identification of INH resistance and clinically relevant NTM are priorities. PMID:24430455

  18. Cellular resistance to HIV-1 infection in target cells coincides with a rapid induction of X-DING-CD4 mRNA: indication of the unique host innate response to virus regulated through function of the X-DING-CD4 gene.

    PubMed

    Shilpi, Rasheda Y; Sachdeva, Rakhee; Simm, Malgorzata

    2012-08-01

    Clinical reports indicate that some infected individuals control HIV-1 replication through undefined mechanisms. Our group reported that a human protein named X-DING-CD4 holds a potent antiviral activity, blocking transcription of HIV-1 LTR through the inhibition of NF-κB/DNA binding. Based on observations that transformed HIV-1 resistant CD4(+) T cells produce higher levels of soluble X-DING-CD4 protein upon their exposure to virus, we hypothesized that resistance to HIV-1 in these cells may be regulated through function of the X-DING-CD4 gene. Real-time PCR evaluations of X-DING-CD4 mRNA expression confirmed our hypothesis; HIV-1 exposure caused rapid up-regulation of X-DING-CD4 mRNA in resistant, but not susceptible, cells; and the burst of X-DING-CD4 mRNA expression correlated with restriction of HIV-1 transcription. Subsequently, we examined the activity of the X-DING-CD4 gene in monocytes and macrophages from (n = 13) HIV-negative donors. The assessment of HIV-1 gag mRNA showed that the majority of cells were permissive to virus replication; however, macrophages from four donors were refractory to HIV-1 infection. In response to virus, these cells up-regulated X-DING-CD4 gene expression by 2- to 1000-fold. These data provide evidence that the X-DING-CD4 gene contributes to early cellular protection from HIV infection in some individuals and this protection depends solely on the unique genetic regulation of the host. PMID:22042911

  19. HIV and Hepatitis Testing: Global Progress, Challenges, and Future Directions.

    PubMed

    Easterbrook, Philippa; Johnson, Cheryl; Figueroa, Carmen; Baggaley, Rachel

    2016-01-01

    HIV infection and viral hepatitis due to HBV and HCV infection are major causes of chronic disease worldwide, and share some common routes of transmission, epidemiology, initial barriers faced in treatment access, and in strategies for a global public health response. Testing and diagnosis of HIV, HBV, and HCV infection is the gateway for access to both care and treatment and prevention services, and crucial for an effective HIV and hepatitis epidemic response. In this review article, we first summarize the common goals and guiding principles in a public health approach to HIV and hepatitis testing. We summarize the impressive global progress in HIV testing scale-up and evolution of approaches, with expansion of provider-initiated testing and counseling in clinical settings (particularly antenatal and tuberculosis clinics), the introduction of more community based testing services, and use of rapid diagnostic tests enabling provision of same-day test results. However, 46% of all people living with HIV are still unaware of their serostatus, and many continue to be diagnosed and start antiretroviral therapy late. As testing and treatment scale-up accelerates for an "treat all" approach, other challenges to address include how to better focus testing and reach those yet undiagnosed and most at risk, especially key populations, men, adolescents, and children. We summarize future directions in HIV testing to speed scale-up and close gaps that are addressed in the WHO 2015 consolidated HIV testing guidelines. In contrast to HIV, action in hepatitis testing and treatment has been fragmented and limited to a few countries, and there remains a large burden of undiagnosed cases globally. We summarize key challenges in the hepatitis testing response, including lack of simple, reliable, and low-cost diagnostic tests, laboratory capacity, and testing facilities; inadequate data to guide country specific hepatitis testing approaches and who to screen; stigmatization and social

  20. Results from a rapid national assessment of services for the prevention of mother-to-child transmission of HIV in Côte d'Ivoire

    PubMed Central

    Granato, S Adam; Gloyd, Stephen; Robinson, Julia; Dali, Serge A; Ahoba, Irma; Aka, David; Kouyaté, Seydou; Billy, Doroux A; Kalibala, Samuel; Koné, Ahoua

    2016-01-01

    Introduction Loss-to-follow-up (LTFU) in the prevention of mother-to-child HIV transmission (PMTCT) programmes can occur at multiple stages of antenatal and follow-up care. This paper presents findings from a national assessment aimed at identifying major bottlenecks in Côte d'Ivoire's PMTCT cascade, and to distinguish characteristics of high- and low-performing health facilities. Methods This cross-sectional study, based on a nationally representative sample of 30 health facilities in Côte d'Ivoire used multiple data sources (registries, patient charts, patient booklets, interviews) to determine the magnitude of LTFU in PMTCT services. A composite measure of retention – based on child prophylaxis, maternal treatment and infant testing – was used to identify high- and low-performing sites and determine significant differences using Student's t-tests. Results Among 1,741 pregnant women newly recorded as HIV-positive between June 2011 and May 2012, 43% had a CD4 count taken, 77% received appropriate prophylaxis and 70% received prophylaxis intended for their infant. During that time, 1,054 first infant HIV tests were recorded. A conservative rate of adherence to antiretroviral therapy was estimated at 50% (n=219 patient charts). Significant differences between high- and low-performing sites included: duration of time elapsed between HIV testing and CD4 results (29.5 versus 56.3 days, p=0.001); and density (number per 100 first antenatal care visits) of full-time physicians (6.7 versus 1.7, p=0.04), laboratory technicians (2.3 versus 0.7, p=0.046), staff trained in PMTCT (10.7 versus 4.7, p=0.01), and staff performing patient follow-up activities (7.9 versus 2.5, p=0.02). Key informants highlighted staff presence and training, the availability of medical supplies and equipment (i.e., on-site CD4 machine), and the adequacy of infrastructure (i.e., space and ventilation) as perceived key factors positively and negatively impacting retention in care. Conclusions

  1. Frequency of HIV Screening in the Veterans Health Administration: Implications for Early Diagnosis of HIV Infection

    ERIC Educational Resources Information Center

    Valdiserri, Ronald O.; Rodriguez, Fred; Holodniy, Mark

    2008-01-01

    We evaluated the frequency of HIV testing across the Department of Veterans Affairs (VA), the largest provider of HIV care in the United States. An electronic survey was used to determine the volume and location of HIV screening, confirmatory testing, rapid testing and laboratory consent policies in VA medical centers between October 1, 2005, and…

  2. A simple and rapid 3D view method for selective and sensitive determination of paclitaxel in micro volume rat plasma by LC-diode array UV and its application to a pharmacokinetic study.

    PubMed

    Kumar, Sekar Vasantha; Srinath, Selladurai; Saha, Ranendra N

    2012-03-01

    A simple, highly repeatable and reproducible method for the estimation of Paclitaxel (TAX) in micro volume amounts of rat plasma is successfully developed and validated. The extraction procedure using 800 µL of ice-cold acetonitrile is very simple and economical with high sensitivity. The rectangular ratiograms and purity curve demonstrate the selectivity of the method. The validation and stability results show that propylparaben (PP) is a suitable internal standard (resolution 7.70 ± 0.15 min) for the estimation of TAX in micro volume rat plasma. TAX and PP are separated by isocratic reversed-phase high-performance liquid chromatography with diode array UV method with a retention time of 8.0 ± 0.25 and 5.3 ± 0.15 min, respectively, with a total run time of 10 min. The system suitability results show that the method has good reproducibility. The stability of TAX is well studied in rat plasma, and the % RSD of all stability studies of TAX are well within the acceptable range of ± 20 % at the lower limit of quantitation (LLOQ) and ± 15% at all quality control levels. The limit of detection (LOD) and LLOQ of the method are 5 and 10 ng/mL, respectively. This rapid method is successfully used to study the i.v pharmacokinetic of TAX at 10 mg/kg in wistar rats, and drug concentration is detected up to 24 h. PMID:22337803

  3. The prevalence and correlates of receiving confirmatory HIV test results among newly diagnosed HIV-positive individuals at a community-based testing center.

    PubMed

    Feldman, Matthew; Wu, Elwin; Mendoza, Moira; Lowry, Blakely; Ford, Lynnette; Holloway, Ian

    2012-10-01

    This study examined the prevalence and correlates of completing the HIV testing process-specifically receiving a confirmatory HIV test and returning for the results-in a sample of newly diagnosed HIV-positive individuals at an HIV testing center in New York City. Of the 213 individuals who received a reactive rapid HIV test result, 82% received a confirmatory HIV test. Of the 236 individuals who received a positive result on a rapid or traditional HIV test that was validated by a positive confirmatory HIV test, 65% returned for the confirmatory test results. Multivariate analyses revealed that being a non-U.S. citizen, homeless/living in transitional housing, being uninsured, and testing off-site were significantly associated with completing the HIV testing process. The findings indicate the need to explore strategies that address obstacles to receiving confirmatory HIV testing and returning for the results, in addition to the feasibility of a rapid confirmatory HIV test. PMID:23016505

  4. Women and HIV

    MedlinePlus

    ... Consumer Information by Audience For Women Women and HIV Share Tweet Linkedin Pin it More sharing options ... HIV? What should pregnant women know about HIV? HIV Quick Facts What is HIV? HIV is the ...

  5. HIV in Southeast Asia.

    PubMed

    Abrams, S

    1998-01-01

    This article explores the HIV/AIDS epidemic in Southeast Asia. Prostitution and injecting drug use are two major factors in the appearance of HIV/AIDS in a country. But, it is the correct social network that assures its transmission to epidemic proportions. Heterosexual transmission in Cambodia, Myanmar, and Thailand is linked with prevalence among female sex workers and their clients. In Malaysia, the Ministry of Health responded immediately, but the number of new infections continued to increase. The failures suggest the need for more effective, intensive health education programs, outreach by nongovernmental organizations, and peer education at the grassroots level and in remote areas. Public health officials need to promote political change. International agencies could play an important role, if countries such as Myanmar, Cambodia, and Viet Nam were open to international exchanges. In Myanmar, political unrest has a priority over the need for aggressive health interventions. In Indonesia, the Islamic influence prevents recognition of the country's significant sex industry or the existence of a homosexual community. In Cambodia, health officials warned about the high number of sexual partners, high mobility rate, and low condom use, but HIV spread rapidly in the 1990s. Thailand initiated a 100% condom campaign to combat HIV prevalence in the 1990s, and HIV prevalence declined among sex workers and military recruits. Risk factors for rapid transmission include mobility, the number of sexual partners/sex worker, the proportion engaging in commercial sex, and the rate of regular condom use among sex workers. PMID:12294443

  6. IDEPI: Rapid Prediction of HIV-1 Antibody Epitopes and Other Phenotypic Features from Sequence Data Using a Flexible Machine Learning Platform

    PubMed Central

    Hepler, N. Lance; Scheffler, Konrad; Weaver, Steven; Murrell, Ben; Richman, Douglas D.; Burton, Dennis R.; Poignard, Pascal; Smith, Davey M.; Kosakovsky Pond, Sergei L.

    2014-01-01

    Since its identification in 1983, HIV-1 has been the focus of a research effort unprecedented in scope and difficulty, whose ultimate goals — a cure and a vaccine – remain elusive. One of the fundamental challenges in accomplishing these goals is the tremendous genetic variability of the virus, with some genes differing at as many as 40% of nucleotide positions among circulating strains. Because of this, the genetic bases of many viral phenotypes, most notably the susceptibility to neutralization by a particular antibody, are difficult to identify computationally. Drawing upon open-source general-purpose machine learning algorithms and libraries, we have developed a software package IDEPI (IDentify EPItopes) for learning genotype-to-phenotype predictive models from sequences with known phenotypes. IDEPI can apply learned models to classify sequences of unknown phenotypes, and also identify specific sequence features which contribute to a particular phenotype. We demonstrate that IDEPI achieves performance similar to or better than that of previously published approaches on four well-studied problems: finding the epitopes of broadly neutralizing antibodies (bNab), determining coreceptor tropism of the virus, identifying compartment-specific genetic signatures of the virus, and deducing drug-resistance associated mutations. The cross-platform Python source code (released under the GPL 3.0 license), documentation, issue tracking, and a pre-configured virtual machine for IDEPI can be found at https://github.com/veg/idepi. PMID:25254639

  7. Rapid and simple determination of acrylamide in conventional cereal-based foods and potato chips through conversion to 3-[bis(trifluoroethanoyl)amino]-3-oxopropyl trifluoroacetate by gas chromatography coupled with electron capture and ion trap mass spectrometry detectors.

    PubMed

    Russo, Mario Vincenzo; Avino, Pasquale; Centola, Angela; Notardonato, Ivan; Cinelli, Giuseppe

    2014-03-01

    A new, simple, rapid and fully validated method based on gas chromatography coupled with Electron capture and ion trap mass spectrometry detectors (GC-ECD and GC-IT/MS) is presented for quantitative analysis of acrylamide contaminant in conventional cereal-based foods and potato chips. Before analysis acrylamide was efficiently derivatized with trifluoroacetic anhydride, the effects of temperature, reaction time and catalyst on the acylation reaction were evaluated. Chromatographic analysis was performed on SE-54 capillary column; good retention and peak response of the acrylamide derivative achieved under the optimal conditions. The analytical method has been fully validated by assessment of the following parameters: LODs and LOQs (1 and 25ngg(-1) by GC-ECD and 2 and 36ngg(-1) by GC-IT/MS, with a Relative Standard Deviations <4 and <6, respectively), linearity (R(2) above 0.981 in the range 0.005-50μgg(-1)) and extraction recovery (ranging between 91% and 99%, RSD below 4.0, for acrylamide spiked at levels of 1, 20, 50 and 100ngg(-1)). Furthermore, the method proposed requires no clean-up step of the acrylamide derivative to be performed prior to injection. The developed method has been successfully applied to determine acrylamide in different commercial cereal-based foods (including French fries and potato chips). PMID:24176333

  8. Finding those at risk: Acute HIV infection in Newark, NJ

    PubMed Central

    Martin, Eugene G.; Salaru, Gratian; Mohammed, Debbie; Coombs, Robert W.; Paul, Sindy M.; Cadoff, Evan M.

    2014-01-01

    Background A screening strategy combining rapid HIV-1/2 (HIV) antibody testing with pooled HIV-1 RNA testing increases identification of HIV infections, but may have other limitations that restrict its usefulness to all but the highest incidence populations. Objective By combining rapid antibody detection and pooled nucleic acid amplification testing (NAAT) testing, we sought to improve detection of early HIV-1 infections in an urban Newark, NJ hospital setting. Study design Pooled NAAT HIV-1 RNA testing was offered to emergency department patients and out-patients being screened for HIV antibodies by fingerstick-rapid HIV testing. For those negative by rapid HIV and agreeing to NAAT testing, pooled plasma samples were prepared and sent to the University of Washington where real-time reverse transcription-polymerase chain reaction (RT-PCR) amplification was performed. Results Of 13,226 individuals screened, 6381 had rapid antibody testing alone, and 6845 agreed to add NAAT HIV screening. Rapid testing identified 115 antibody positive individuals. Pooled NAAT increased HIV-1 case detection by 7.0% identifying 8 additional cases. Overall, acute HIV infection yield was 0.12%. While males represent only 48.1% of those tested by NAAT, all samples that screened positive for HIV-1 RNA were obtained from men. Conclusion HIV-1 RNA testing of pooled, HIV antibody-negative specimens permits identification of recent infections. In Newark, pooled NAAT increased HIV-1 case detection and provided an opportunity to focus on treatment and prevention messages for those most at risk of transmitting infection. Although constrained by client willingness to participate in testing associated with a need to return to receive further results, use of pooled NAAT improved early infection sensitivity. PMID:23953941

  9. HIV Surveillance in a Large, Community-Based Study: Results from the Pilot Study of Project Accept (HIV Prevention Trials Network 043)

    PubMed Central

    2011-01-01

    Background Project Accept is a community randomized, controlled trial to evaluate the efficacy of community mobilization, mobile testing, same-day results, and post-test support for the prevention of HIV infection in Thailand, Tanzania, Zimbabwe, and South Africa. We evaluated the accuracy of in-country HIV rapid testing and determined HIV prevalence in the Project Accept pilot study. Methods Two HIV rapid tests were performed in parallel in local laboratories. If the first two rapid tests were discordant (one reactive, one non-reactive), a third HIV rapid test or enzyme immunoassay was performed. Samples were designated HIV NEG if the first two tests were non-reactive, HIV DISC if the first two tests were discordant, and HIV POS if the first two tests were reactive. Samples were re-analyzed in the United States using a panel of laboratory tests. Results HIV infection status was correctly determined based on-in country testing for 2,236 (99.5%) of 2,247 participants [7 (0.37%) of 1,907 HIV NEG samples were HIV-positive; 2 (0.63%) of 317 HIV POS samples were HIV-negative; 2 (8.3%) of 24 HIV DISC samples were incorrectly identified as HIV-positive based on the in-country tie-breaker test]. HIV prevalence was: Thailand: 0.6%, Tanzania: 5.0%, Zimbabwe 14.7%, Soweto South Africa: 19.4%, Vulindlela, South Africa: 24.4%, (overall prevalence: 14.4%). Conclusions In-country testing based on two HIV rapid tests correctly identified the HIV infection status for 99.5% of study participants; most participants with discordant HIV rapid tests were not infected. HIV prevalence varied considerably across the study sites (range: 0.6% to 24.4%). Trial Registration ClinicalTrials.gov registry number NCT00203749. PMID:21943026

  10. Rapid infectious diseases diagnostics using Smartphones

    PubMed Central

    Bates, Matthew

    2015-01-01

    The “Smartphone” is an almost universal possession in high-income populations, and is rapidly becoming so in lower-income regions, particularly among urban populations, and serves social networking and a quest for information and knowledge. The field of infectious disease diagnostics is at a potential watershed moment, with the essential building blocks for the development of diagnostic assays being ever more available and affordable, which is leading to creative innovative approaches to developing much-needed accurate and simple point-of-care (POC) diagnostic tools for high disease burden, low-income settings. We review the importance and implications of a paper published in Science Translational Medicine on the development of a smartphone-powered and -controlled multiplex immunological assay that tests for HIV and syphilis simultaneously. This is reviewed in the context of other prototype smartphone-enabled/assisted diagnostic devices, and how such developments might shape the future of the POC diagnostics field. PMID:26488011

  11. HIV-1 Capsid Stabilization Assay.

    PubMed

    Fricke, Thomas; Diaz-Griffero, Felipe

    2016-01-01

    The stability of the HIV-1 core in the cytoplasm is crucial for productive HIV-1 infection. Mutations that stabilize or destabilize the core showed defects in HIV-1 reverse transcription and infection. We developed a novel and simple assay to measure stability of in vitro-assembled HIV-1 CA-NC complexes. This assay allowed us to demonstrate that cytosolic extracts strongly stabilize the HIV-1 core (Fricke et al., J Virol 87:10587-10597, 2013). By using our novel assay, one can measure the ability of different drugs to modulate the stability of in vitro-assembled HIV-1 CA-NC complexes, such as PF74, CAP-1, IXN-053, cyclosporine A, Bi2, and the peptide CAI. We also found that purified CPSF6 (1-321) protein stabilizes in vitro-assembled HIV-1 CA-NC complexes (Fricke et al., J Virol 87:10587-10597, 2013). Here we describe in detail the use of this capsid stability assay. We believe that our assay can be a powerful tool to assess HIV-1 capsid stability in vitro. PMID:26714703

  12. Get Tested for HIV

    MedlinePlus

    ... Print This Topic En español Get Tested for HIV Browse Sections The Basics Overview What Is HIV? ... 1 of 7 sections The Basics: What Is HIV? What is HIV? HIV stands for human immunodeficiency ...

  13. HIV Treatment: The Basics

    MedlinePlus

    HIV Treatment HIV Treatment: The Basics (Last updated 3/1/2016; last reviewed 3/1/2016) Key Points Antiretroviral therapy (ART) ... reduces the risk of HIV transmission . How do HIV medicines work? HIV attacks and destroys the infection- ...

  14. Simple scale interpolator facilitates reading of graphs

    NASA Technical Reports Server (NTRS)

    Fetterman, D. E., Jr.

    1965-01-01

    Simple transparent overlay with interpolation scale facilitates accurate, rapid reading of graph coordinate points. This device can be used for enlarging drawings and locating points on perspective drawings.

  15. Cost-Effective and Rapid Presumptive Identification of Gram-Negative Bacilli in Routine Urine, Pus, and Stool Cultures: Evaluation of the Use of CHROMagar Orientation Medium in Conjunction with Simple Biochemical Tests

    PubMed Central

    Ohkusu, Kiyofumi

    2000-01-01

    The algorithm for a new identification system was designed on the basis of colony color and morphology on CHROMagar Orientation medium in conjunction with simple biochemical tests such as indole (IND), lysine decarboxylase (LDC), and ornithine decarboxylase (ODC) utilization tests with gram-negative bacilli isolated from urine samples as well as pus, stool, and other clinical specimens by the following colony characteristics, biochemical reactions, and serological results: pinkish to red, IND positive (IND+), Escherichia coli; metallic blue, IND+, LDC+, and ODC negative (ODC−), Klebsiella oxytoca; IND+, LDC−, and ODC+, Citrobacter diversus; IND+ or IND−, LDC−, and ODC−, Citrobacter freundii; IND−, LDC+, and ODC+, Enterobacter aerogenes; IND−, LDC−, and ODC+, Enterobacter cloacae; IND−, LDC+, and ODC−, Klebsiella pneumoniae; diffuse brown and IND+, Morganella morganii; IND−, Proteus mirabilis; aqua blue, Serratia marcescens; bluish green and IND+, Proteus vulgaris; transparent yellow-green, serology positive, Pseudomonas aeruginosa; clear and serology positive, Salmonella sp.; other colors and reactions, the organism was identified by the full identification methods. The accuracy and cost-effectiveness of this new system were prospectively evaluated. During an 8-month period, a total of 345 specimens yielded one or more gram-negative bacilli. A total of 472 gram-negative bacillus isolates were detected on CHROMagar Orientation medium. For 466 of the isolates (98.7%), no discrepancies in the results were obtained on the basis of the identification algorithm. The cost of identification of gram-negative bacilli during this period was reduced by about 70%. The results of this trial for the differentiation of the most commonly encountered gram-negative pathogens in clinical specimens with the new algorithm were favourable in that it permitted reliable detection and presumptive identification. In addition, this rapid identification system not only

  16. Screening for acute HIV infection in South Africa: finding acute and chronic disease

    PubMed Central

    Bassett, Ingrid V.; Chetty, Senica; Giddy, Janet; Reddy, Shabashini; Bishop, Karen; Lu, Zhigang; Losina, Elena; Freedberg, Kenneth A.; Walensky, Rochelle P.

    2010-01-01

    Background The yield of screening for acute HIV infection among general medical patients in resource-scarce settings remains unclear. Our objective was to evaluate a strategy of pooled HIV plasma RNA to diagnose acute HIV infection in patients with negative or discordant rapid HIV antibody tests in Durban, South Africa. Methods We prospectively enrolled patients with negative or discordant rapid HIV antibody tests from a routine HIV screening program in an outpatient department in Durban with an HIV prevalence of 48%. Study participants underwent venipuncture for pooled qualitative HIV RNA, and if positive, quantitative RNA, enzyme immunoassay and Western Blot (WB). Patients with negative or indeterminate WB and positive quantitative HIV RNA were considered acutely infected. Those with chronic infection (positive RNA and WB) despite negative or discordant rapid HIV tests were considered false negative rapid antibody tests. Results Nine hundred ninety-four participants were enrolled with either negative (N=976) or discordant (N=18) rapid test results. Eleven (1.1%, 95% CI: 0.6–2.0%) had acute HIV infection. Of the 994 patients, an additional 20 (2.0%, 95% CI: 1.3–.3.1%) had chronic HIV infection (false negative rapid test). Conclusions One percent of outpatients with negative or discordant rapid HIV tests in Durban, South Africa had acute HIV infection readily detectable through pooled serum HIV RNA screening. Pooled RNA testing also identified an additional 2% of patients with chronic HIV infection. HIV RNA screening has the potential to identify both acute and chronic HIV infections that are otherwise missed by standard HIV testing algorithms. PMID:20553336

  17. Contextual factors influencing HIV risk behavior in Central Asia

    PubMed Central

    Smolak, Alex

    2010-01-01

    Central Asia has experienced a rapid increase in HIV. HIV interventions and prevention programmes are needed that adequately appreciate and account for the ways that ongoing cultural, political, and economic changes in this region affect HIV risk reduction efforts. Drawing on relevant literature, this paper provides a contextual foundation to better understand the impact of context on HIV risk behaviour in the countries of Central Asia and to begin the conversation on the contextual factors of Islam and polygamy. PMID:20301020

  18. HIV prevention transformed: the new prevention research agenda.

    PubMed

    Padian, Nancy S; McCoy, Sandra I; Karim, Salim S Abdool; Hasen, Nina; Kim, Julia; Bartos, Michael; Katabira, Elly; Bertozzi, Stefano M; Schwartländer, Bernhard; Cohen, Myron S

    2011-07-16

    We have entered a new era in HIV prevention whereby priorities have expanded from biomedical discovery to include implementation, effectiveness, and the effect of combination prevention at the population level. However, gaps in knowledge and implementation challenges remain. In this Review we analyse trends in the rapidly changing landscape of HIV prevention, and chart a new path for HIV prevention research that focuses on the implementation of effective and efficient combination prevention strategies to turn the tide on the HIV pandemic. PMID:21763938

  19. Advances in Developing HIV-1 Viral Load Assays for Resource-Limited Settings

    PubMed Central

    Wang, ShuQi; Xu, Feng; Demirci, Utkan

    2010-01-01

    Commercial HIV-1 RNA viral load assays have been routinely used in developed countries to monitor antiretroviral treatment (ART). However, these assays require expensive equipment and reagents, well-trained operators, and established laboratory infrastructure. These requirements restrict their use in resource-limited settings where people are most afflicted with the HIV-1 epidemic. Inexpensive alternatives such as the Ultrasensitive p24 assay, the Reverse Transcriptase (RT) assay and in-house reverse transcription quantitative polymerase chain reaction (RT-qPCR) have been developed. However, they are still time-consuming, technologically complex and inappropriate for decentralized laboratories as point-of-care (POC) tests. Recent advances in microfluidics and nanotechnology offer new strategies to develop low-cost, rapid, robust and simple HIV-1 viral load monitoring systems. We review state-of-the-art technologies used for HIV-1 viral load monitoring in both developed and developing settings. Emerging approaches based on microfluidics and nanotechnology, which have potential to be integrated into POC HIV-1 viral load assays, are also discussed. PMID:20600784

  20. eHealth interventions for HIV prevention.

    PubMed

    Noar, Seth M; Willoughby, Jessica Fitts

    2012-01-01

    The rapidly changing media landscape and proliferation of new technologies creates vast new opportunities for HIV prevention. The fast growth of the relatively new eHealth field is a testament to the excitement and promise of these new technologies. eHealth interventions in HIV prevention tested to date include computer- and Internet-based interventions; chat room interventions; text messaging interventions; and social media. The current article provides a brief review of these types of interventions in HIV prevention, including their unique advantages and evidence of efficacy. Implications for future research in the eHealth HIV prevention field are discussed. PMID:22519523

  1. HIV surveillance in complex emergencies.

    PubMed

    Salama, P; Dondero, T J

    2001-04-01

    Many studies have shown a positive association between both migration and temporary expatriation and HIV risk. This association is likely to be similar or even more pronounced for forced migrants. In general, HIV transmission in host-migrant or host-forced-migrant interactions depends on the maturity of the HIV epidemic in both the host and the migrant population, the relative seroprevalence of HIV in the host and the migrant population, the prevalence of other sexually transmitted infections (STIs) that may facilitate transmission, and the level of sexual interaction between the two communities. Complex emergencies are the major cause of mass population movement today. In complex emergencies, additional factors such as sexual interaction between forced-migrant populations and the military; sexual violence; increasing commercial sex work; psychological trauma; and disruption of preventive and curative health services may increase the risk for HIV transmission. Despite recent success in preventing HIV infection in stable populations in selected developing countries, internally displaced persons and refugees (or forced migrants) have not been systematically included in HIV surveillance systems, nor consequently in prevention activities. Standard surveillance systems that rely on functioning health services may not provide useful data in many complex emergency settings. Secondary sources can provide some information in these settings. Little attempt has been made, however, to develop innovative HIV surveillance systems in countries affected by complex emergencies. Consequently, data on the HIV epidemic in these countries are scarce and HIV prevention programs are either not implemented or interventions are not effectively targeted. Second generation surveillance methods such as cross-sectional, population-based surveys can provide rapid information on HIV, STIs, and sexual behavior. The risks for stigmatization and breaches of confidentiality must be recognized

  2. HIV Transmission

    MedlinePlus

    ... pre-chewed by an HIV-infected person. The contamination occurs when infected blood from a caregiver’s mouth ... pre-chewed by an HIV-infected person. The contamination occurs when infected blood from a caregiver’s mouth ...

  3. Human Microbiome and HIV/AIDS

    PubMed Central

    Li, Yihong; Yang, Liying; Pei, Zhiheng; Poles, Michael; Abrams, William R.; Malamud, Daniel

    2013-01-01

    Understanding of the human microbiome continues to grow rapidly; however, reports on changes in the microbiome after HIV infection are still limited. This review surveys the progress made in methodology associated with microbiome studies and highlights the remaining challenges to this field. Studies have shown that commensal oral, gut, vaginal, and penile bacteria are vital to the health of the human immune system. Our studies on crosstalk among oral and gastrointestinal soluble innate factors, HIV, and microbes indicated that the oral and gut microbiome was altered in the HIV-positive samples compared to the negative controls. The importance of understanding the bacterial component of HIV/AIDS, and likelihood of “crosstalk” between viral and bacterial pathogens, will help in understanding the role of the microbiome in HIV-infected individuals and facilitate identification of novel antiretroviral factors for use as novel diagnostics, microbicides, or therapeutics against HIV infection. PMID:22193889

  4. Oral lesions, HIV phenotypes, and management of HIV-related disease: Workshop 4A.

    PubMed

    Patton, L L; Ranganathan, K; Naidoo, S; Bhayat, A; Balasundaram, S; Adeyemi, O; Taiwo, O; Speicher, D J; Chandra, L

    2011-04-01

    The workshop considered 5 questions related to oral lesions, HIV phenotypes, and the management of HIV-related disease, with a focus on evidence and challenges in resource-poor settings. First, are oral lesions unique with respect to geographic location or phenotype? Second, how useful would an oral lesion index be to predict HIV in resource-poor countries with no access to CD4 counts or viral load? Third, what are the latest methods and delivery modes for drugs used to treat oral lesions associated with HIV? Fourth, what is the role of the oral health care worker in rapid diagnostic testing for HIV? Fifth, what ethical and legal issues are to be considered when managing the HIV patient? The consensus of the workshop was the need for additional research in 4 key areas in developing countries: (1) additional investigation of comorbidities associated with HIV infection that may affect oral lesion presentation and distribution, especially in pediatric populations; (2) the development of region-specific algorithms involving HIV oral lesions, indicating cumulative risk of immune suppression and the presence of HIV disease; (3) well-designed clinical trials to test new therapies for oral lesions, new treatments for resistant oral fungal and viral diseases, effectiveness of therapies in children, and new drug delivery systems; and (4) the role of the oral health care worker in rapid diagnostic testing for HIV in various regions of the world. PMID:21441491

  5. HIV Medication Adherence

    MedlinePlus

    HIV Treatment HIV Medication Adherence (Last updated 3/1/2016; last reviewed 3/1/2016) Key Points Medication adherence means sticking ... exactly as prescribed. Why is adherence to an HIV regimen important? Adherence to an HIV regimen gives ...

  6. HIV among Transgender People

    MedlinePlus

    ... of transgender Virginians . Richmond, VA: Virginia HIV Community Planning Committee and Virginia Department of Health; 2007. Accessed April 14, 2016. Additional ... HIV/AIDS CDC HIV CDC HIV/AIDS ...

  7. Screening and diagnosis for HIV

    MedlinePlus

    HIV testing; HIV screening; HIV screening test; HIV confirmatory test ... A positive result on a screening test does not confirm that the person has HIV infection. More tests are needed to confirm HIV infection. A negative test ...

  8. Preventing sexual transmission of HIV: anti-HIV bioregulatory and homeostatic components of commercial sexual lubricants.

    PubMed

    Nguyen, D; Lee, H; Poast, J; Cloyd, M W; Baron, S

    2004-01-01

    Certain safe over-the-counter (OTC) sexual lubricants such as Astroglide, KY Liquid, Replens, Vagisil, ViAmor, and Wet Stuff inhibit both cell-free HIV and the production of HIV by infected leukocytes in vitro even in the presence of seminal fluid. To identify which components of the lubricants were active against HIV, we tested five components (glycerin, methylparaben, propylparaben, polyquaternium-32, and propylene glycol). The paraben preservatives and propylene glycol in the lubricants did not inhibit HIV, while the common natural homeostatic metabolite, glycerin, and the thickener polyquaternium-32 did strongly inactivate infectious HIV and HIV-infected leukocytes. Activity against HIV and HIV-infected cells by glycerin was stable through 24 hours at 37 degrees C. Glycerin and polyquaternium-32 were active at minimum concentrations of approximately 2% and 0.01%, respectively--well within the highest FDA safety guidelines. Both active components disrupted infected leukocytes within 5 minutes which resulted in inhibition of infectious HIV production by infected leukocytes of greater than 25 to 100-fold. These components do not disrupt vaginal epithelial cells in vivo. These components also rapidly inactivate cell-free HIV by 10- to 30-fold. Thus, we may conclude that the active components of the OTC lubricants are glycerin and polyquaternium-32. Using these components, OTC sexual lubricants could be reformulated to optimize their anti-HIV activity. Furthermore, clinical trials of these lubricants and components seem to be indicated because of their FDA safety level, wide availability, and low cost. PMID:15786693

  9. HIV among Women

    MedlinePlus

    ... testing site near you. Enter ZIP code or city Follow HIV/AIDS CDC HIV CDC HIV/AIDS ... HIV infection—National HIV Behavioral Surveillance, 20 U.S. cities, 2013 . HIV Surveillance Special Report 13 . Accessed January ...

  10. Maternal HIV status affects the infant hemoglobin level

    PubMed Central

    Feleke, Berhanu Elfu

    2016-01-01

    Abstract Children, especially infants, are highly vulnerable to iron-deficiency anemia because of their rapid growth of the brain and the rest of the body. The objectives of this study were to compare the prevalence of iron-deficiency anemia in infants born from HIV-positive mothers and HIV-negative mothers and to identify the determinants of iron-deficiency anemia in infants. A comparative cross-sectional study was conducted in Bahir Dar city. Simple random sampling technique was used to select the study participants. Mothers were interviewed; blood samples were collected from mothers and infants to measure the hemoglobin level and anthropometric indicators were obtained from the infants using world health organization standards. Descriptive statistics were used to estimate the prevalence of infantile anemia. Binary logistic regression and multiple linear regressions were used to identify the determinants of infant anemia. A total of 1459 infants born from HIV-positive and HIV-negative mothers were included. The prevalence of iron-deficiency anemia in infants born from HIV-positive and HIV-negative mothers was 41.9% (95% CI: 39–44). Infantile iron-deficiency anemia was associated with maternal HIV infection (adjusted odds ratio [AOR] 2.54 [95% CI: 1.65–3.9]), stunting (AOR 3.46 [95% CI: 2.41–4.97]), low income (AOR 2.72 [95% CI: 2–3.73]), maternal malaria during pregnancy (AOR 1.81 [95% CI: 1.33–2.47]), use of cow milk before 6 month (AOR 1.82 [95% CI: 1.35–2.45]), residence (AOR 0.09 [95% CI: 0.06–0.13]), history of cough or fever 7 days preceding the survey (AOR 2.71 [95% CI: 1.99–3.69]), maternal hemoglobin (B 0.65 [95% CI: 0.61–0.68]), educational status of mother (B 0.22 [95% CI: 0.2–0.23]), age of the mother (B –0.03 [95% CI: –0.03, –0.02]), and family size (B –0.14 [95% CI: –0.18,–0.11]). PMID:27495044

  11. A cost-effective sandwich electrochemiluminescence immunosensor for ultrasensitive detection of HIV-1 antibody using magnetic molecularly imprinted polymers as capture probes.

    PubMed

    Zhou, Jing; Gan, Ning; Li, Tianhua; Hu, Futao; Li, Xing; Wang, Lihong; Zheng, Lei

    2014-04-15

    In this report, a rapid and cost-effective sandwich electrochemiluminescence (ECL) immunosensor was constructed for the ultrasensitive detection of human immunodeficiency virus type 1 antibody (anti-HIV-1) using magnetic molecularly imprinted polymers (MMIPs) as capture probes by combining surface and epitope imprinting techniques and antigen conjugated with horseradish peroxidase (HRP-HIV-1) as labels. First, 3-aminobenzeneboronic acid (APBA) was used as the functional monomer and cross-linking reagent, which was polymerized on the surface of silicate-coated magnetic iron oxide nanoparticles (Fe3O4@SiO2 NPs) in the presence of human immunoglobulin G (HIgG), as the template exhibiting the same Fc region but different Fab region to anti-HIV-1 after the addition of the initiator, ammonium persulfate. This process resulted in grafting a hydrophilic molecularly imprinted polymer (MIP) film on the Fe3O4@SiO2 NPs. Thus, MMIPs, which could be reused after eluting the template, were used to recognize and enrich ultra-trace levels of anti-HIV-1. Subsequently, a novel sandwich ECL immunosensor was formed through the immunoreaction between MMIPs conjugated with varied concentrations of anti-HIV-1 and HRP-HIV-1. By the catalysis of HRP immobilized onto HRP-HIV-1 on the ECL system of Luminol-H2O2, a linear response range of the anti-HIV-1 dilution ratio (standard positive serum) was achieved from 1:20,000 to 1:50, with a detection limit of 1:60,000 (S/N=3). The developed method provides a low-cost, simple, and sensitive way for the early diagnosis of HIV infected patients. PMID:24280050

  12. HIV / AIDS

    MedlinePlus

    ... Marketing Share this: Main Content Area Understanding HIV/AIDS AIDS was first reported in the United States in ... and has since become a major worldwide epidemic. AIDS is caused by the human immunodeficiency virus, or ...

  13. Humanized Mouse Models of HIV Infection

    PubMed Central

    Denton, Paul W.; Garcia, J. Victor

    2013-01-01

    Because of the limited tropism of HIV, in vivo modeling of this virus has been almost exclusively limited to other lentiviruses such as SIV that reproduce many important characteristics of HIV infection. However, there are significant genetic and biological differences among lentiviruses and some HIV-specific interventions are not effective against other lentiviruses in non-human hosts. For these reasons much emphasis has recently been placed on developing alternative animal models that support HIV replication and recapitulate key aspects of HIV infection and pathogenesis in humans. Humanized mice, CD34+ hematopoietic progenitor cell transplanted immunodeficient mice and in particular mice also implanted with human thymic/liver tissue (BLT mice) that develop a functional human immune system, have been the focus of a great deal of attention as possible models to study virtually all aspects of HIV biology and pathogenesis. Humanized mice are systemically reconstituted with human lymphoid cells offering rapid, reliable and reproducible experimental systems for HIV research. Peripheral blood of humanized mice can be readily sampled longitudinally to assess reconstitution with human cells and to monitor HIV replication permitting the evaluation of multiple parameters of HIV infection such as viral load levels, CD4+ T cell depletion, immune activation, as well as the effects of therapeutic interventions. Of high relevance to HIV transmission is the extensive characterization and validation of the reconstitution with human lymphoid cells of the female reproductive tract and of the gastrointestinal tract of humanized BLT mice that renders them susceptible to both vaginal and rectal HIV infection. Other important attributes of all types of humanized mice include: 1) their small size and cost that make them broadly accessible; 2) multiple cohorts of humanized mice can be made from multiple human donors and each cohort has identical human cells, permitting control of

  14. Effectiveness of the U.S. National HIV Testing Day Campaigns in Promoting HIV Testing: Evidence from CDC-Funded HIV Testing Sites, 2010

    PubMed Central

    Mulatu, Mesfin S.

    2014-01-01

    Objectives We assessed if HIV testing and diagnoses increased during the week of National HIV Testing Day (NHTD) and if characteristics of people who were tested varied compared with control weeks. Methods We analyzed HIV testing data from the 2010 National HIV Prevention Program Monitoring and Evaluation system to compare NHTD week (June 24–30, 2010) with two control weeks (January 7–13, 2010, and August 12–18, 2010) for the number of HIV testing events and new HIV-positive diagnoses, by demographics and other HIV-related variables. Characteristics associated with testing during NHTD week compared with control weeks were identified using Chi-square analyses. Results In 2010, an average of 15,000 more testing events were conducted and 100 more new HIV-positive diagnoses were identified during NHTD week than during the control weeks (p<0.001). Compared with control weeks, people tested during NHTD week were significantly less likely to be aged 20–29 years and non-Hispanic white and significantly more likely to be (1) aged ≥50 years, (2) non-Hispanic black or African American, (3) men who have sex with men, (4) low-risk heterosexuals, (5) tested with a rapid HIV test, or (6) tested in a non-health-care setting. Conclusion In 2010, CDC-funded HIV testing events and new HIV-positive diagnoses increased during NHTD week compared with control weeks. HIV testing programs increased the use of rapid tests and returned a high percentage of test results. NHTD campaigns reached populations disproportionately affected by HIV and further expanded testing to people traditionally less likely to be tested. Incorporating strategies used during NHTD in programs conducted throughout the year may assist in increasing HIV testing and the number of HIV-positive diagnoses. PMID:25177056

  15. Simple manipulator for rotating spheres.

    PubMed

    Weinstein, B W; Hendricks, C D; Ward, C M; Willenborg, D L

    1978-06-01

    We describe a simple device for rapidly rotating a small sphere to any orientation for inspection of the surface. The ball is held between two small, flat surfaces and rolls as the surfaces are moved differentially parallel to one another. PMID:18699214

  16. Atazanavir/ritonavir-based combination antiretroviral therapy for treatment of HIV-1 infection in adults.

    PubMed

    Achenbach, Chad J; Darin, Kristin M; Murphy, Robert L; Katlama, Christine

    2011-02-01

    In the past 15 years, improvements in the management of HIV infection have dramatically reduced morbidity and mortality. Similarly, rapid advances in antiretroviral medications have resulted in the possibility of life-long therapy with simple and tolerable regimens. Protease inhibitors have been important medications in regimens of combination antiretroviral therapy for the treatment of HIV. One of the recommended and commonly used therapies in this class is once-daily-administered atazanavir, pharmacologically boosted with ritonavir (atazanavir/r). Clinical studies and practice have shown these drugs, in combination with other antiretroviral agents, to be potent, safe and easy to use in a variety of settings. Atazanavir/r has minimal short-term toxicity, including benign bilirubin elevation, and has less potential for long-term complications of hyperlipidemia and insulin resistance compared with other protease inhibitors. A high genetic barrier to resistance and a favorable resistance profile make it an excellent option for initial HIV treatment or as the first drug utilized in the protease inhibitors class. Atazanavir/r is also currently being studied in novel treatment strategies, including combinations with new classes of antiretrovirals to assess nucleoside reverse transcriptase inhibitor-sparing regimens. In this article we review atazanavir/r as a treatment for HIV infection and discuss the latest information on its pharmacology, efficacy and toxicity. PMID:21731578

  17. Atazanavir/ritonavir-based combination antiretroviral therapy for treatment of HIV-1 infection in adults

    PubMed Central

    Achenbach, Chad J; Darin, Kristin M; Murphy, Robert L; Katlama, Christine

    2011-01-01

    In the past 15 years, improvements in the management of HIV infection have dramatically reduced morbidity and mortality. Similarly, rapid advances in antiretroviral medications have resulted in the possibility of life-long therapy with simple and tolerable regimens. Protease inhibitors have been important medications in regimens of combination antiretroviral therapy for the treatment of HIV. One of the recommended and commonly used therapies in this class is once-daily-administered atazanavir, pharmacologically boosted with ritonavir (atazanavir/r). Clinical studies and practice have shown these drugs, in combination with other antiretroviral agents, to be potent, safe and easy to use in a variety of settings. Atazanavir/r has minimal short-term toxicity, including benign bilirubin elevation, and has less potential for long-term complications of hyperlipidemia and insulin resistance compared with other protease inhibitors. A high genetic barrier to resistance and a favorable resistance profile make it an excellent option for initial HIV treatment or as the first drug utilized in the protease inhibitors class. Atazanavir/r is also currently being studied in novel treatment strategies, including combinations with new classes of antiretrovirals to assess nucleoside reverse transcriptase inhibitor-sparing regimens. In this article we review atazanavir/r as a treatment for HIV infection and discuss the latest information on its pharmacology, efficacy and toxicity. PMID:21731578

  18. The cytokine network in HIV infection.

    PubMed

    Alfano, M; Poli, G

    2002-12-01

    Cytokines are major controller of HIV replication and represent, at the same time, a target for viral-induced immune dysregulation. This mutual relationship has profound implications for both active HIV replication and immune-mediated governance of latency; in addition, cytokines have therapeutic value in the perspective of immune reconstitution. In the current article we will review the most relevant aspects emerged in almost 20 years of research in this area with particular reference to the distinct, but interconnected contribution of the most simple (cell lines) to the most complex (animal) models of HIV infection. PMID:12462389

  19. [Pneumocystosis during HIV infection].

    PubMed

    El Fane, M; Sodqi, M; Oulad Lahsen, A; Chakib, A; Marih, L; Marhoum El Filali, K

    2016-08-01

    Pneumocystosis is an opportunistic disease caused by invasion of unicellular fungus Pneumocystic jirovecii which is responsible for febrile pneumonia among patients with cellular immunodeficiency especially those HIV infected. Despite the decreasing of its incidence due to the introduction of antiretroviral therapy, as well as anti-Pneumocystis prophylaxis among these patients, Pneumocystis pneumonia remains the first AIDS-defining event and a leading cause of mortality among HIV-infected patients. The usual radiological presentation is that of diffuse interstitial pneumonia. The diagnosis is confirmed by the detection of trophozoides and/or cysts P. jirovecii in bronchoalveolar lavage (BAL) samples using several staining techniques. The use of polymerase chain reaction in the BAL samples in conjunction with standard immunofluorescent or colorimetric tests have allowed for more has allowed for more rapid and accurate diagnosis. The standard regimen of treatment is the association of trimethoprim-sulfamethoxazole which has been utilized as an effective treatment with a favourable recovery. Early HIV diagnosis and antiretroviral therapy should reduce the incidence of this dreaded disease. PMID:27349824

  20. Simple, efficient UHV manipulator

    SciTech Connect

    Thiel, P.A.; Anderegg, J.W.

    1984-10-01

    A simple manipulator is described for use in ultrahigh vacuum. Unhindered rotation within vacuum is provided by a commercial differentially pumped adapter to which the entire XYZ stage is attached. The combination of movements provided by the XYZ stage, the rotating vacuum flange, and a gimbal permits alignment of the sample with any of the peripheral view ports. The resistively heated sample support, mounted through a cold finger, permits rapid variation of sample temperature from several hundred degrees Kelvin to near cryogenic. It is anticipated that this design could be easily used with many existing types of commercial vacuum systems, with the consequent advantage of increased ease and simplicity of both mechanical motion and cryogenic cooling.

  1. HIV Life Cycle

    MedlinePlus

    HIV Overview The HIV Life Cycle (Last updated 9/8/2016; last reviewed 9/8/2016) Key Points HIV gradually destroys the immune ... life cycle. What is the connection between the HIV life cycle and HIV medicines? Antiretroviral therapy (ART) ...

  2. Nanomaterials and Optical Diagnosis of HIV.

    PubMed

    Valizadeh, Alireza

    2016-09-01

    The investigators had previously shown that the risk of AIDS/HIV-related illness and transmission reduced (by 96%) with early antiretroviral treatment. Nanomaterials could be applied in early diagnosis of HIV by improving the ability to detect serum biomarkers of the blood-borne infectious diseases, with low sample volume, rapidity, and more sensitivity than currently available FDA-approved methods such as ELISA, particle agglutination assay, and Western Blotting assay. We have demonstrated several experimental studies for optical HIV diagnosis based on nanomaterials in three categories (e.g., the fluorescence-, the SPR-, and the SERS- based biosensors), and have explained each assay. PMID:26099718

  3. Comparison of viro-immunological marker changes between HIV-1 and HIV-2-infected patients in France

    PubMed Central

    Drylewicz, Julia; Matheron, Sophie; Lazaro, Estibaliz; Damond, Florence; Bonnet, Fabrice; Simon, François; Dabis, François; Brun-Vezinet, Françoise; Chêne, Geneviève; Thiebaut, Rodolphe

    2008-01-01

    Background HIV-2 is known to be less pathogenic than HIV-1, although the underlying mechanisms are still debated. We compared the changes over time in viro-immunologic markers in HIV-1 and HIV-2 infected patients living in France during natural history and after initiation of the first Combination of AntiRetroviral Treatment (CART). Method Patients were included in the ANRS CO3 HIV-1 cohort (N=6707) or the ANRS CO5 HIV-2 cohort (N=572). HIV-1 infected patients were matched to HIV-2 patients according to sex, age, HIV transmission group and period of treatment initiation. Changes in markers have been estimated with linear mixed models. Results Analyses were performed for three groups of patients: (1) those with estimated date of contamination (98 HIV-1 and 49 HIV-2 seroincident patients), (2) untreated seroprevalent patients (320 HIV-1 and 160 HIV-2) and (3) those who initiated a first CART (59 HIV-1 and 63 HIV-2). In group 1, CD4 T-cell decreased less rapidly in HIV-2 than HIV-1 patients (−9 vs. −49 cells/mm3/year, p<10−4). Estimated slopes in untreated group 2 were similar to those estimated in group 1 (−11 vs. −49 cells/mm3/year, p=0.003). In group 3, baseline CD4 at CART initiation was not different according to the type of infection (269 vs.220 cells/mm3). During the first two months of treatment, CD4 count increased by +59 cells/mm3/month (95% Confidence Interval [CI]=34;84) for HIV-1 and +24 (CI=6;42) for HIV-2. The plasma viral load drop was 3-fold more important in HIV-1 patients: −1.56 log10/ml/month (CI=−1.83; − 1.30) vs. −0.62 (CI=−0.84; −0.40) among HIV-2 patients (p<10−4). Conclusion Differences between the two infections during natural history are similar to those previously described in Africa. Paradoxically, once treatment is started, response is poorer in HIV-2 patients than in HIV-1 patients. PMID:18301058

  4. The Invention Convention: Mind Meets Simple Machines.

    ERIC Educational Resources Information Center

    Hadi-Tabassum, Samina

    1997-01-01

    Describes an Earth Day celebration where students had to design an invention made of simple machines that could crush an empty aluminum can through 10 rapid mechanical movements using materials foraged from the students' homes. (JRH)

  5. Mucosal Immunology of HIV Infection

    PubMed Central

    Xu, Huanbin; Wang, Xiaolei; Veazey, Ronald S.

    2013-01-01

    Summary Recent advances in the immunology, pathogenesis, and prevention of human immunodeficiency virus (HIV) infection continue to reveal clues to the mechanisms involved in the progressive immunodeficiency attributed to infection but more importantly have shed light on the correlates of immunity to infection and disease progression. HIV selectively infects, eliminates, and/or dysregulates several key cells of the human immune system, thwarting multiple arms of the host immune response, and inflicting severe damage to mucosal barriers, resulting in tissue infiltration of ‘symbiotic’ intestinal bacteria and viruses that essentially become opportunistic infections promoting systemic immune activation. This leads to activation and recruitment or more target cells for perpetuating HIV infection, resulting in persistent, high level viral replication in lymphoid tissues, rapid evolution of resistant strains, and continued evasion of immune responses. However, vaccine studies and studies of spontaneous controllers are finally providing correlates of immunity from protection and disease progression, including virus-specific CD4+ T-cell responses, binding antibodies, innate immune responses, and generation of antibodies with potent antibody-dependent cell-mediated cytotoxicity activity. Emerging correlates of immunity indicate that prevention of HIV infection may be possible through effective vaccine strategies that protect and stimulate key regulatory cells and immune responses in susceptible hosts. Further, immune therapies specifically directed towards boosting specific aspects of the immune system may eventually lead to a cure for HIV-infected patients. PMID:23772612

  6. Representational Fluency in HIV Clinical Practice: A Model of Instructor Discourse

    ERIC Educational Resources Information Center

    Banach, Mary A.; Gifford, Bernard R.; Holodniy, Mark

    2007-01-01

    Introduction: Clinicians treating human immunodeficiency virus (HIV) patients are expected to stay up-to-date with rapidly changing knowledge and practice. Continuing medical education (CME) programs are one source of new knowledge about HIV clinical management. Little is known about instructor-participant discourse in HIV CME programs and whether…

  7. HIV Testing

    MedlinePlus

    ... the right way, every day. If you have health insurance, your insurer is required to cover some medicines ... to treat HIV. If you don’t have health insurance, or you’re unable to afford your co- ...

  8. HIV/AIDS

    MedlinePlus

    ... at risk for serious infections and certain cancers. AIDS stands for acquired immunodeficiency syndrome. It is the final stage of infection with HIV. Not everyone with HIV develops AIDS. HIV most often spreads through unprotected sex with ...

  9. Preventing HIV with Medicine

    MedlinePlus

    ... information in Spanish ( en español ) Preventing HIV with medicine Get medicine right after you are exposed to ... to top More information on Preventing HIV with medicine Explore other publications and websites National HIV and ...

  10. HIV/AIDS

    MedlinePlus

    HIV stands for human immunodeficiency virus. It kills or damages the body's immune system cells. AIDS stands for acquired immunodeficiency syndrome. It is the most advanced stage of infection with HIV. HIV most ...

  11. How HIV Causes AIDS

    MedlinePlus

    ... Share this: Main Content Area How HIV Causes AIDS HIV destroys CD4 positive (CD4+) T cells, which ... and disease, ultimately resulting in the development of AIDS. Most people who are infected with HIV can ...

  12. HIV/AIDS Basics

    MedlinePlus

    ... Enter ZIP code or city Follow Act Against AIDS Act Against AIDS @talkHIV Act Against AIDS Get Email Updates on AAA Anonymous Feedback HIV/AIDS Media Infographics Syndicated Content Podcasts Slide Sets HIV/ ...

  13. HIV-AIDS Connection

    MedlinePlus

    ... Marketing Share this: Main Content Area The HIV-AIDS Connection AIDS was first recognized in 1981 and ... is there overwhelming scientific consensus that HIV causes AIDS? Before HIV infection became widespread in the human ...

  14. Older People and HIV

    MedlinePlus

    ... common than they were before the use of anti-HIV drugs. It is difficult to know what is causing mental problems in older people with HIV. Is it normal aging, or is it HIV disease? Research studies have ...

  15. Integrating HIV Care and HIV Prevention: Legal, Policy, and Programmatic Recommendations

    PubMed Central

    Remien, Robert H.; Berkman, Alan; Myer, Landon; Bastos, Francisco I.; Kagee, Ashraf; El-Sadr, Wafaa

    2009-01-01

    Since the start of the HIV epidemic we have witnessed significant advances in our understanding of the impact of HIV disease worldwide. Further, breakthroughs in treatment and the rapid expansion of HIV care and treatment programs in heavily impacted countries over the past five years are potentially critical assets in a comprehensive approach to controlling the continued spread of HIV globally. A strategic approach to controlling the epidemic requires continued and comparable expansion and integration of care, treatment, and prevention programs. As every new infection involves transmission, whether vertically or horizontally, from a person already living with HIV/AIDS (PLWHA), integration of HIV prevention into HIV care settings has the potential to prevent thousands of new infections, as well as improve the lives of PLWHAs. In this paper, we highlight how to better utilize opportunities created by the antiretroviral (ARV) roll-out to achieve more effective prevention, particularly in Sub Saharan Africa. We offer specific recommendations for action in the domains of healthcare policy and practice in order to better utilize the advances in HIV treatment to advance HIV prevention. PMID:18641470

  16. Sexual Risk Behaviors for HIV/AIDS in Chuuk State, Micronesia: The Case for HIV Prevention in Vulnerable Remote Populations

    PubMed Central

    Russell, Toya V.; Do, Ann N.; Setik, Eleanor; Sullivan, Patrick S.; Rayle, Victoria D.; Fridlund, Carol A.; Quan, Vu M.; Voetsch, Andrew C.; Fleming, Patricia L.

    2007-01-01

    Background After the first two cases of locally-acquired HIV infection were recognized in Chuuk State, Federated States of Micronesia (FSM), a public health response was initiated. The purpose of the response was to assess the need for HIV education and prevention services, to develop recommendations for controlling further spread of HIV in Chuuk, and to initiate some of the prevention measures. Methodology/Principal Findings A public health team conducted a survey and rapid HIV testing among a sample of residents on the outer islands in Chuuk. Local public health officials conducted contact tracing and testing of sex partners of the two locally-acquired cases of HIV infection. A total of 333 persons completed the survey. The majority knew that HIV is transmitted through unprotected sexual contact (81%), injection drug use (61%), or blood transfusion (64%). Sexual activity in the past 12 months was reported among 159 participants, including 90 females and 69 males. Compared to women, men were more likely to have had multiple sex partners, to have been drunk during sex, but less likely to have used a condom in the past 12 months. The two men with locally acquired HIV infection had unprotected anal sex with a third Chuukese man who likely contracted HIV while outside of Chuuk. All 370 persons who received voluntary, confidential HIV counseling and testing had HIV negative test results. Conclusions/Significance Despite the low HIV seroprevalence, risky sexual behaviors in this small isolated population raise concerns about the potential for rapid spread of HIV. The lack of knowledge about risks, along with stigmatizing attitudes towards persons infected with HIV and high risk sexual behaviors indicate the need for resources to be directed toward HIV prevention in Chuuk and on other Pacific Islands. PMID:18074009

  17. A rapid and simple method for the simultaneous determination of four endogenous monoamine neurotransmitters in rat brain using hydrophilic interaction liquid chromatography coupled with atmospheric-pressure chemical ionization tandem mass spectrometry.

    PubMed

    Zhou, Wenbin; Zhu, Bangjie; Liu, Feng; Lyu, Chunming; Zhang, Shen; Yan, Chao; Cheng, Yu; Wei, Hai

    2015-10-01

    Endogenous monoamine neurotransmitters play an essential role in neural communication in mammalians. Many quantitative methods for endogenous monoamines have been developed during recent decades. Yet, matrix effect was usually a challenge in the quantification, in many cases asking for tedious sample preparation or sacrificing sensitivity. In this work, a simple, fast and sensitive method with no matrix effect was developed to simultaneously determine four endogenous monoamines including serotonin, dopamine, epinephrine and norepinephrine in rat brain tissues, using hydrophilic interaction liquid chromatography coupled with atmospheric-pressure chemical ionization tandem mass spectrometry. Various conditions, including columns, chromatographic conditions, ion source, MS/MS conditions, and brain tissue preparation methods, were optimized and validated. Pre-weighed 20mg brain sample could be effectively and reproducibly homogenized and protein-precipitated by 20 times value of 0.2% formic acid in cold organic solvents (methanol-acetonitrile, 10:90, v/v). This method exhibited excellent linearity for all analytes (regression coefficients>0.998 or 0.999). The precision, expressed as coefficients of variation, was less than 3.43% for intra-day analyses and ranged from 4.17% to 15.5% for inter-day analyses. Good performance was showed in limit of detection (between 0.3nM and 3.0nM for all analytes), recovery (90.8-120%), matrix effect (84.4-107%), accuracy (89.8-100%) and stability (88.3-104%). The validated method was well applied to simultaneously determine the endogenous serotonin, dopamine, epinephrine and norepinephrine in four brain sections of 18 Wistar rats. The quantification of four endogenous monoamines in rat brain performed excellently in the sensitivity, high throughput, simple sample preparation and matrix effect. PMID:26363373

  18. NELA: A Community Response to HIV/AIDS in Nigeria

    ERIC Educational Resources Information Center

    Soyinka, Femi; Ogundare, Dipo; Olowookere, Kemi; Akinsola, Yemisi; Alade, Adeyemi; Moronkola, O. A.

    2004-01-01

    The greatest current threat to humanity, most especially in the developing countries of the world, is HIV/AIDS. The first case of HIV/AIDS in Nigeria was in 1986 in Lagos. Due to inaction and denial by the people, there was a rapid but subtle transmission of the virus within Nigeria's various populations and communities. Presently, the disease has…

  19. Improving HIV proteome annotation: new features of BioAfrica HIV Proteomics Resource

    PubMed Central

    Druce, Megan; Hulo, Chantal; Masson, Patrick; Sommer, Paula; Xenarios, Ioannis; Le Mercier, Philippe; De Oliveira, Tulio

    2016-01-01

    The Human Immunodeficiency Virus (HIV) is one of the pathogens that cause the greatest global concern, with approximately 35 million people currently infected with HIV. Extensive HIV research has been performed, generating a large amount of HIV and host genomic data. However, no effective vaccine that protects the host from HIV infection is available and HIV is still spreading at an alarming rate, despite effective antiretroviral (ARV) treatment. In order to develop effective therapies, we need to expand our knowledge of the interaction between HIV and host proteins. In contrast to virus proteins, which often rapidly evolve drug resistance mutations, the host proteins are essentially invariant within all humans. Thus, if we can identify the host proteins needed for virus replication, such as those involved in transporting viral proteins to the cell surface, we have a chance of interrupting viral replication. There is no proteome resource that summarizes this interaction, making research on this subject a difficult enterprise. In order to fill this gap in knowledge, we curated a resource presents detailed annotation on the interaction between the HIV proteome and host proteins. Our resource was produced in collaboration with ViralZone and used manual curation techniques developed by UniProtKB/Swiss-Prot. Our new website also used previous annotations of the BioAfrica HIV-1 Proteome Resource, which has been accessed by approximately 10 000 unique users a year since its inception in 2005. The novel features include a dedicated new page for each HIV protein, a graphic display of its function and a section on its interaction with host proteins. Our new webpages also add information on the genomic location of each HIV protein and the position of ARV drug resistance mutations. Our improved BioAfrica HIV-1 Proteome Resource fills a gap in the current knowledge of biocuration. Database URL: http://www.bioafrica.net/proteomics/HIVproteome.html PMID:27087306

  20. Improving HIV proteome annotation: new features of BioAfrica HIV Proteomics Resource.

    PubMed

    Druce, Megan; Hulo, Chantal; Masson, Patrick; Sommer, Paula; Xenarios, Ioannis; Le Mercier, Philippe; De Oliveira, Tulio

    2016-01-01

    The Human Immunodeficiency Virus (HIV) is one of the pathogens that cause the greatest global concern, with approximately 35 million people currently infected with HIV. Extensive HIV research has been performed, generating a large amount of HIV and host genomic data. However, no effective vaccine that protects the host from HIV infection is available and HIV is still spreading at an alarming rate, despite effective antiretroviral (ARV) treatment. In order to develop effective therapies, we need to expand our knowledge of the interaction between HIV and host proteins. In contrast to virus proteins, which often rapidly evolve drug resistance mutations, the host proteins are essentially invariant within all humans. Thus, if we can identify the host proteins needed for virus replication, such as those involved in transporting viral proteins to the cell surface, we have a chance of interrupting viral replication. There is no proteome resource that summarizes this interaction, making research on this subject a difficult enterprise. In order to fill this gap in knowledge, we curated a resource presents detailed annotation on the interaction between the HIV proteome and host proteins. Our resource was produced in collaboration with ViralZone and used manual curation techniques developed by UniProtKB/Swiss-Prot. Our new website also used previous annotations of the BioAfrica HIV-1 Proteome Resource, which has been accessed by approximately 10 000 unique users a year since its inception in 2005. The novel features include a dedicated new page for each HIV protein, a graphic display of its function and a section on its interaction with host proteins. Our new webpages also add information on the genomic location of each HIV protein and the position of ARV drug resistance mutations. Our improved BioAfrica HIV-1 Proteome Resource fills a gap in the current knowledge of biocuration.Database URL:http://www.bioafrica.net/proteomics/HIVproteome.html. PMID:27087306

  1. A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China.

    PubMed

    Chai, Zheng; Ma, Wenjun; Fu, Fang; Lang, Yuekun; Wang, Wei; Tong, Guangzhi; Liu, Qinfang; Cai, Xuehui; Li, Xi

    2013-02-01

    SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82.98 ± 0.25 °C and 85.95 ± 0.24 °C for HP-PRRSVs or 82.74 ± 0.26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID(50) for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China. PMID:23070137

  2. A novel highly flexible, simple, rapid and low-cost fabrication tool for paper-based microfluidic devices (μPADs) using technical drawing pens and in-house formulated aqueous inks.

    PubMed

    Nuchtavorn, Nantana; Macka, Mirek

    2016-05-01

    Paper-based microfluidic devices (μPADs) are capable of achieving rapid quantitative measurements of a variety of analytes inexpensively. μPADs rely on patterning hydrophilic-hydrophobic regions on a sheet of paper in order to create capillary channels within impermeable fluidic brakes on the paper. Here, we present a novel, highly flexible and low-cost fabrication method using a desktop digital craft plotter/cutter and technical drawing pens with tip size of 0.5 mm. The pens were used with either commercial black permanent ink for drawing fluidic brakes, or with specialty in-house formulated aqueous inks. With the permanent marker ink it was possible to create barriers on paper rapidly and in a variety of designs in a highly flexible manner. For instance, a design featuring eight reservoirs can be produced within 10 s for each μPAD with a consistent line width of brakes (%RSD < 1.5). Further, we investigated the optimal viscosity range of in-house formulated inks controlled with additions of poly(ethylene glycol). The viscosity was measured by capillary electrophoresis and the optimal viscosity was in the range of ∼3-6 mPa s. A functional test of these μPADs was conducted by the screening of antioxidant activity. Colorimetric measurements of flavonoid, phenolic compounds and DPPH free radical scavenging activity were carried out on μPADs. The results can be detected by the naked eye and simply quantified by using a camera phone and image analysis software. The fabrication method using technical drawing pens provides flexibility in the use of in-house formulated inks, short fabrication time, simplicity and low cost. PMID:27086101

  3. Development and Validation of an HIV Risk Exposure and Indicator Conditions Questionnaire to Support Targeted HIV Screening

    PubMed Central

    Elías, María Jesús Pérez; Gómez-Ayerbe, Cristina; Elías, Pilar Pérez; Muriel, Alfonso; de Santiago, Alberto Diaz; Martinez-Colubi, María; Moreno, Ana; Santos, Cristina; Polo, Lidia; Barea, Rafa; Robledillo, Gema; Uranga, Almudena; Espín, Agustina Cano; Quereda, Carmen; Dronda, Fernando; Casado, Jose Luis; Moreno, Santiago

    2016-01-01

    Abstract The aim of our study was to develop a Spanish-structured HIV risk of exposure and indicator conditions (RE&IC) questionnaire. People attending to an emergency room or to a primary clinical care center were offered to participate in a prospective, 1 arm, open label study, in which all enrolled patients filled out our developed questionnaire and were HIV tested. Questionnaire accuracy, feasibility, and reliability were evaluated. Valid paired 5329 HIV RE&IC questionnaire and rapid HIV tests were performed, 69.3% in the primary clinical care center, 49.6% women, median age 37 years old, 74.9% Spaniards, 20.1% Latin-Americans. Confirmed hidden HIV infection was detected in 4.1%, while HIV RE&IC questionnaire was positive in 51.2%. HIV RE&IC questionnaire sensitivity was 100% to predict HIV infection, with a 100% negative predictive value. When considered separately, RE or IC items sensitivity decreases to 86.4% or 91%, and similarly their negative predictive value to 99.9% for both of them. The majority of people studied, 90.8% self-completed HIV RE&IC questionnaire. Median time to complete was 3 minutes. Overall HIV RE&IC questionnaire test-retest Kappa agreement was 0.82 (almost perfect), likewise for IC items 0.89, while for RE items was lower 0.78 (substantial). A feasible and reliable Spanish HIV RE&IC self questionnaire accurately discriminated all non–HIV-infected people without missing any HIV diagnoses, in a low prevalence HIV infection area. The best accuracy and reliability were obtained when combining HIV RE&IC items. PMID:26844471

  4. Development and Validation of an HIV Risk Exposure and Indicator Conditions Questionnaire to Support Targeted HIV Screening.

    PubMed

    Elías, María Jesús Pérez; Gómez-Ayerbe, Cristina; Elías, Pilar Pérez; Muriel, Alfonso; de Santiago, Alberto Diaz; Martinez-Colubi, María; Moreno, Ana; Santos, Cristina; Polo, Lidia; Barea, Rafa; Robledillo, Gema; Uranga, Almudena; Espín, Agustina Cano; Quereda, Carmen; Dronda, Fernando; Casado, Jose Luis; Moreno, Santiago

    2016-02-01

    The aim of our study was to develop a Spanish-structured HIV risk of exposure and indicator conditions (RE&IC) questionnaire. People attending to an emergency room or to a primary clinical care center were offered to participate in a prospective, 1 arm, open label study, in which all enrolled patients filled out our developed questionnaire and were HIV tested. Questionnaire accuracy, feasibility, and reliability were evaluated.Valid paired 5329 HIV RE&IC questionnaire and rapid HIV tests were performed, 69.3% in the primary clinical care center, 49.6% women, median age 37 years old, 74.9% Spaniards, 20.1% Latin-Americans. Confirmed hidden HIV infection was detected in 4.1%, while HIV RE&IC questionnaire was positive in 51.2%. HIV RE&IC questionnaire sensitivity was 100% to predict HIV infection, with a 100% negative predictive value. When considered separately, RE or IC items sensitivity decreases to 86.4% or 91%, and similarly their negative predictive value to 99.9% for both of them. The majority of people studied, 90.8% self-completed HIV RE&IC questionnaire. Median time to complete was 3 minutes. Overall HIV RE&IC questionnaire test-retest Kappa agreement was 0.82 (almost perfect), likewise for IC items 0.89, while for RE items was lower 0.78 (substantial).A feasible and reliable Spanish HIV RE&IC self questionnaire accurately discriminated all non-HIV-infected people without missing any HIV diagnoses, in a low prevalence HIV infection area. The best accuracy and reliability were obtained when combining HIV RE&IC items. PMID:26844471

  5. HIV-1 assembly in macrophages

    PubMed Central

    2010-01-01

    The molecular mechanisms involved in the assembly of newly synthesized Human Immunodeficiency Virus (HIV) particles are poorly understood. Most of the work on HIV-1 assembly has been performed in T cells in which viral particle budding and assembly take place at the plasma membrane. In contrast, few studies have been performed on macrophages, the other major target of HIV-1. Infected macrophages represent a viral reservoir and probably play a key role in HIV-1 physiopathology. Indeed macrophages retain infectious particles for long periods of time, keeping them protected from anti-viral immune response or drug treatments. Here, we present an overview of what is known about HIV-1 assembly in macrophages as compared to T lymphocytes or cell lines. Early electron microscopy studies suggested that viral assembly takes place at the limiting membrane of an intracellular compartment in macrophages and not at the plasma membrane as in T cells. This was first considered as a late endosomal compartment in which viral budding seems to be similar to the process of vesicle release into multi-vesicular bodies. This view was notably supported by a large body of evidence involving the ESCRT (Endosomal Sorting Complex Required for Transport) machinery in HIV-1 budding, the observation of viral budding profiles in such compartments by immuno-electron microscopy, and the presence of late endosomal markers associated with macrophage-derived virions. However, this model needs to be revisited as recent data indicate that the viral compartment has a neutral pH and can be connected to the plasma membrane via very thin micro-channels. To date, the exact nature and biogenesis of the HIV assembly compartment in macrophages remains elusive. Many cellular proteins potentially involved in the late phases of HIV-1 cycle have been identified; and, recently, the list has grown rapidly with the publication of four independent genome-wide screens. However, their respective roles in infected cells

  6. A new idea for simple and rapid monitoring of gene expression: requirement of nucleotide sequences encoding an N-terminal HA tag in the T7 promoter-driven expression in E. coli.

    PubMed

    Moon, Jeong-Mi; Kim, Goo-Young; Rhim, Hyangshuk

    2012-10-01

    Mammalian expression vectors are used to overexpress genes of interest in mammalian cells. High temperature requirement protein A1 (HtrA1), used as a specific target, was expressed from the pHA-M-HtrA1 plasmid in HEK293T cells, inducing cell death. Expression of HtrA1 was driven by the pHA-M-HtrA1 mammalian expression vector in E. coli resulting in growth suppression of E. coli in an HtrA1 serine protease-dependent manner. By using various combinations of promoters, target genes and N-terminal tags, the T7 promoter and N-terminal HA tag in the mammalian expression vector were shown to be responsible for expression of target genes in E. coli. Thus the pHA-M-HtrA1 plasmid can be used as a novel, rapid pre-test system for expression and cytotoxicity of the specific target gene in E. coli before assessing its functions in mammalian cells. PMID:22714269

  7. Unique variability of tocopherol composition in various seed oils recovered from by-products of apple industry: rapid and simple determination of all four homologues (α, β, γ and δ) by RP-HPLC/FLD.

    PubMed

    Górnaś, Paweł

    2015-04-01

    The tocochromanol profile was studied in seed oils recovered from by-products of fruit industry, five dessert and seven crab apple varieties grown in Eastern Europe (Latvia). The seed oils obtained from dessert apples were characterized by higher contents of tocopherols (191.05-379.08 mg/100g oil) when compared to seed oils recovered from crab apples (130.55-202.54 mg/100g oil). The predominant homologues of tocopherol in all the studied samples were α and β over γ and δ. However, seed oils recovered from the apple cultivars 'Antej' and 'Beforest' had a unique profile of four tocopherol homologues (α:β:γ:δ) 91.41:80.55:72.46:79.03 and 114.55:112.84:78.69:73.00 mg/100g oil, respectively. A single dilution of seed oils in 2-propanol facilitated the direct use samples in the DPPH assay as well as injection into the RP-HPLC system containing a PFP (pentafluorophenyl) column, which resulted in a rapid separation of all four tocopherol homologues with excellent repeatability and reproducibility. PMID:25442533

  8. Chlamydia trachomatis Infection in HIV-Infected Women: Need for Screening by a Sensitive and Specific Test

    PubMed Central

    Bhattar, Sonali; Chadha, Sanjim; Tripathi, Reva; Kaur, Ravinder; Sardana, Kabir

    2013-01-01

    Reproductive tract infection (RTIs)/sexually transmitted infections (STIs) are recognized as a major public health problem, particularly due to their relationship with HIV infection. Early detection and treatment of Chlamydia trachomatis infection (CTI) among HIV-infected and HIV-uninfected women may impact heterosexual HIV transmission. A total of 120 participants were enrolled: 30 HIV seropositive women with symptoms of RTIs, 30 HIV seropositive women without symptoms of RTIs, 30 HIV seronegative women with symptoms of RTIs, and 30 HIV seronegative women without symptoms of RTIs. One endocervical swab was collected from all participants and CTI was detected by real-time PCR (COBAS TaqMan CT Test, v2.0). CTI was detected in 4 (6.67%) HIV-infected women and in 1 (1.67%) HIV-uninfected woman (OR 4.214; 95% CI 0.457–38.865). Vaginal discharge was present in almost half of HIV-infected and HIV-uninfected women; lower abdominal pain was present in 11 (18.3%) of HIV-infected and in 9 (15%) of HIV-uninfected women. This study showed that CTI is more prevalent among HIV-infected females as compared to HIV-uninfected females. As the use of real-time PCR is not feasible in most hospitals, efforts should be made to develop a simple, sensitive, and specific test to identify women with CTI for prevention of sequelae and HIV transmission. PMID:24382941

  9. Performance of an alternative HIV diagnostic algorithm using the ARCHITECT HIV Ag/Ab Combo assay and potential utility of sample-to-cutoff ratio to discriminate primary from established infection☆

    PubMed Central

    Ramos, Eric M.; Harb, Socorro; Dragavon, Joan; Swenson, Paul; Stekler, Joanne D.; Coombs, Robert W.

    2014-01-01

    Background The ARCHITECT HIV Ag/Ab Combo assay has a wide dynamic range for determining the sample-to-cutoff ratio (S/CO) values compared to other diagnostic HIV antibody assays. Objectives Determine the performance of an HIV testing algorithm that uses the ARCHITECT combo assay in the clinical setting and explore the utility of the signal-to-cutoff (S/CO) ratio to predict acute HIV-1 infection status. Study design A retrospective analysis of clinical samples from a hospital and referral population screened for HIV-1 infection between May 2011 and March 2013. Repeatedly reactive samples were tested using the Multispot HIV-1/HIV-2 rapid test and depending on that result, confirmatory orthogonal testing used the Western blot (WB) for HIV-1, Immunoblot for HIV-2 and nucleic acid amplification testing (NAAT) for HIV RNA. Results A total of 21,317 test results were evaluated of which 509 were ARCHITECT repeatedly reactive; of these, 422 were Multispot-reactive only for HIV-1 (413 WB-positive; 9 indeterminate), 4 were Multispot-reactive for both HIV-1 and HIV-2 (one HIV-2 immunoblot-positive with 17 HIV-2 RNA copies/mL) and 83 were Multispot-non-reactive of which 15 were HIV-1 RNA positive and represented acute HIV-1 infection. There was an association among the ARCHITECT S/CO (median; IQR) values for antibody-negative (0.14; 0.11–0.16), acute infection (33; 2.1–76) and established HIV-1 infection (794; 494–1,029) (Kruskal–Wallis, p < 0.0001). Conclusions The ARCHITECT combo assay with Multispot confirmation and reserved use of HIV-1 WB, HIV-2 Immunoblot and HIV NAAT for Multispot dual HIV-1/2 infection, and NAAT alone for Multispot-negative specimens, had a suitable test performance for detecting acute and established HIV infection. PMID:24029686

  10. HIV prevention transformed: the new prevention research agenda

    PubMed Central

    Padian, Nancy S.; McCoy, Sandra I.; Karim, Salim Abdool; Hasen, Nina; Kim, Julia; Bartos, Michael; Katabira, Elly; Bertozzi, Stefano; Schwartländer, Bernhard; Cohen, Myron S.

    2013-01-01

    SUMMARY We have entered a new era in HIV prevention whereby priorities have expanded from biomedical discovery to include implementation, effectiveness, and the effect of combination prevention at the population level. However, gaps in knowledge and implementation challenges remain. In this Review we analyse trends in the rapidly changing landscape of HIV prevention, and chart a new path for HIV prevention research that focuses on the implementation of effective and efficient combination prevention strategies to turn the tide on the HIV pandemic. PMID:21763938

  11. Interrelationship of alcohol misuse, HIV sexual risk and HIV screening uptake among emergency department patients

    PubMed Central

    2013-01-01

    Background Emergency department (ED) patients comprise a high-risk population for alcohol misuse and sexual risk for HIV. In order to design future interventions to increase HIV screening uptake, we examined the interrelationship among alcohol misuse, sexual risk for HIV and HIV screening uptake among these patients. Methods A random sample of 18-64-year-old English- or Spanish-speaking patients at two EDs during July-August 2009 completed a self-administered questionnaire about their alcohol use using the Alcohol Use Questionnaire, the Alcohol Use Disorders Identification Test (AUDIT), and the HIV Sexual Risk Questionnaire. Study participants were offered a rapid HIV test after completing the questionnaires. Binging (≥ five drinks/occasion for men, ≥ four drinks for women) was assessed and sex-specific alcohol misuse severity levels (low-risk, harmful, hazardous, dependence) were calculated using AUDIT scores. Analyses were limited to participants who had sexual intercourse in the past 12 months. Multivariable logistic regression was used to assess the associations between HIV screening uptake and (1) alcohol misuse, (2) sexual risk for HIV, and (3) the intersection of HIV sexual risk and alcohol misuse. Adjusted odds ratios (AORs) with 95% confidence intervals (CIs) were estimated. All models were adjusted for patient demographic characteristics and separate models for men and women were constructed. Results Of 524 participants (55.0% female), 58.4% identified as white, non-Hispanic, and 72% reported previous HIV testing. Approximately 75% of participants reported drinking alcohol within the past 30 days and 74.5% of men and 59.6% of women reported binge drinking. A relationship was found between reported sexual risk for HIV and alcohol use among men (AOR 3.31 [CI 1.51-7.24]) and women (AOR 2.78 [CI 1.48-5.23]). Women who reported binge drinking were more likely to have higher reported sexual risk for HIV (AOR 2.55 [CI 1.40-4.64]) compared to women who do

  12. A Simple and Rapid Method Based on Liquid Chromatography-Tandem Mass Spectrometry for the Measurement of α-L-Iduronidase Activity in Dried Blood Spots: An Application to Mucopolysaccharidosis I (Hurler) Screening

    PubMed Central

    Yang, Jeong Soo; Min, Hye Kyeong; Oh, Hyeon Ju; Woo, Hye In; Lee, Soo-Youn; Kim, Jong-Won; Song, Junghan

    2015-01-01

    Background We developed an analytical method to measure α-L-iduronidase (IDUA) activity in dried blood spots. This was achieved by using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) with electrospray ionization in the positive ion mode. Methods Chromatographic separation was completed using mobile phase involving water-formic acid and acetonitrile-formic acid over 2.8 min of run time on a column with a Kinetex XB-C18 (Phenomenex, USA). The detection of column effluent was performed using a Xevo TQ-S triple quadrupole mass spectrometer (Waters, USA) in the multiple-reaction monitoring mode. This method was verified with blank and control samples at four activity levels: base, low, medium, and high. Control materials were provided from Centers for Disease Control and Prevention (CDC). Results Intra- and inter-day precisions were between 2.6% and 16.5% and between 7.9% and 17.0%, respectively. A correlative regression study on the IDUA activity in CDC-control samples performed to assess the validity of the developed method showed a highly significant linear association (r2=0.9976) between the calculated and CDC-reported values and an obvious difference in activity among the four levels. This reliable analytical method was applied to mucopolysaccharidosis I (Hurler) screening of patients under treatment (n=4) and in normal controls (n=129). IDUA activity ranged from 8.98 to 77.12 µmol/hr/L) in normal controls, and patients undergoing medical treatment showed low IDUA activity. Conclusions This method had advantages of simplicity, rapid sample preparation, and liquid chromatographic separation, which efficiently inhibited ionization suppression induced by matrix effects in mass spectrometric detection. PMID:25553279

  13. Simple Kidney Cysts

    MedlinePlus

    ... Organizations​​ . (PDF, 345 KB)​​​​​ Alternate Language URL Simple Kidney Cysts Page Content On this page: What are simple ... Points to Remember Clinical Trials What are simple kidney cysts? Simple kidney cysts are abnormal, fluid-filled sacs ...

  14. HIV and AIDS

    MedlinePlus

    ... I Help a Friend Who Cuts? HIV and AIDS KidsHealth > For Teens > HIV and AIDS Print A A A Text Size What's in ... in human history. HIV causes a condition called acquired immunodeficiency syndrome — better known as AIDS . HIV destroys a type ...

  15. HIV Structural Database

    National Institute of Standards and Technology Data Gateway

    SRD 102 HIV Structural Database (Web, free access)   The HIV Protease Structural Database is an archive of experimentally determined 3-D structures of Human Immunodeficiency Virus 1 (HIV-1), Human Immunodeficiency Virus 2 (HIV-2) and Simian Immunodeficiency Virus (SIV) Proteases and their complexes with inhibitors or products of substrate cleavage.

  16. An SNP marker at the STAT6 locus can identify the hybrids between rhesus (Macaca mulatta) and long-tailed macaques (M. fascicularis) in Thailand: a rapid and simple screening method and its application.

    PubMed

    Jadejaroen, Janya; Kawamoto, Yoshi; Hamada, Yuzuru; Malaivijitnond, Suchinda

    2016-01-01

    A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was developed to genetically discriminate rhesus (Macaca mulatta) macaques from long-tailed (M. fascicularis) macaques. The 745 bp PCR amplicon of the STAT6 locus that spans a potentially species-diagnostic single nucleotide polymorphism (SNP) marker was digested with ApaI and gel electrophoresed to give (1) two (234 and 511 bp), (2) one (745 bp) and (3) three (234, 511 and 745 bp) band patterns that correspond to the genotypes G/G (long-tailed macaque specific homozygote), A/A (rhesus macaque specific homozygote) and A/G (hybrid specific heterozygote), respectively. The diagnostic robustness and efficiency of this PCR-RFLP assay was tested on wild rhesus and long-tailed macaques inhabiting Thailand and a known hybrid population. The Indochinese and Sundaic long-tailed macaque samples (n = 18) all showed a homozygous G/G pattern, while the Indochinese rhesus macaques (n = 10) all showed a homozygous A/A pattern. The rhesus/long-tailed hybrid population at Khao Khieow Open Zoo, which resulted from an introduced group of rhesus macaques that hybridized with the indigenous long-tailed macaques about 20 years ago, revealed 47% (56/118 samples analyzed) with the heterogenous A/G genotype. In addition, the frequency of the rhesus-specific allele A significantly decreased in the hybrid population during 2006-2014, where a strong association between the STAT6 genotype and the morphology of the individuals was detected. In conclusion, a robust PCR-RFLP assay allows a simple, effective and inexpensive approach, in particular for field studies, to assess hybrid individuals between rhesus and long-tailed macaques. Although this assay cannot conclusively identify all the hybrids over two or more generations, it at least can allow the evaluation of the process of hybridization, and so it is applicable to the assessment of the status of natural or anthropogenic hybridization between the two

  17. Mutagenicity and pausing of HIV reverse transcriptase during HIV plus-strand DNA synthesis.

    PubMed Central

    Ji, J; Hoffmann, J S; Loeb, L

    1994-01-01

    The unusually high frequency of misincorporation by HIV-1 reverse transcriptase (HIV RT) is likely to be the major factor in the rapid accumulation of viral mutations in AIDS, especially in the env gene. To investigate the ability of HIV RT to copy the env gene, we subcloned an HIV env gene fragment into a single-stranded DNA vector and measured the progression of synthesis by HIV RT. We observed that HIV RT, but not RT from avian myeloblastosis virus, DNA polymerase-alpha or T7 DNA polymerase, pauses specifically at poly-deoxyadenosine stretches within the env gene. The frequency of bypassing the polyadenosine stretches by HIV RT is enhanced by increasing the ratio of enzyme to template. We measured the fidelity of DNA synthesis within a segment of the hypervariable region 1 of the env gene (V-1) containing a poly-deoxyadenosine sequence by repetitively copying the DNA by HIV RT, and then cloning and sequencing the copied fragments. We found that 27% of the errors identified in V-1 sequence were frameshift mutations opposite the poly-adenosine tract, a site where strong pausing was observed. Pausing of HIV RT at the polyadenosine tract could be enhanced by either distamycin A or netropsin, (A-T)-rich minor groove binding peptides. Moreover, netropsin increases the frequency of frameshift mutations in experiments in which HIV RT catalyzes gap filling synthesis within the lacZ gene in double-stranded circular M13mp2 DNA. These combined results suggest that the enhanced mutation frequency may be due to increased pausing at netropsin-modified polyadenosine tracts. Therefore, netropsin and related A-T binding chemicals may selectively enhance frameshift mutagenesis induced by HIV RT and yield predominantly non-viable virus. Images PMID:7510388

  18. Testing (HIV). Quick test receives Singapore approval.

    PubMed

    1996-04-22

    Hema-Strip HIV 1/2 is a rapid HIV antibody immunoassay developed by Saliva Diagnostic Systems, Inc. (SDS) which can be used by anyone who can read the product insert. The test kit is comprised of a small lancet for a finger stick, a cylindrical tube with a capillary tip and a SDS diagnostic strip inside, and a vial of buffer. Once blood is drawn by the lancet, the capillary tip is placed upon the blood droplet and the blood is automatically drawn into the tube. The tube is then inserted tip first into the vial of buffer. The buffer and blood migrate over the diagnostic strip inside, yielding stable results within 15 minutes. Studies have found Hema-Strip HIV 1/2 to have a sensitivity and specificity greater than 99.4%, as accurate as most conventional HIV tests which require the use of laboratory equipment and trained staff, and possibly hours to produce results. Moreover, the test kit requires neither refrigeration nor special storage. Hema-Strip HIV 1/2 has received a certificate of free sale from the Ministry of Health in Singapore and is now being submitted for regulatory approval in Brazil, China, Russia, India, Malaysia, Thailand, and the UK. SDS products in production include Sero-Strip HIV 1/2, a rapid serum-based HIV antibody test; Omni-SAL, a saliva collector which is the principal sample collection device used by British insurance companies for HIV testing with other confirmatory tests; Omni-Swab, a serrated swab which collects body fluids or cells; Saliva-Sampler, a saliva collection device used for general testing purposes; and Saliva Check, a test which checks the composition of saliva samples. SDS is in the final stages of developing Saliva-Strip HIV-1/2, a rapid saliva-based HIV antibody test. The company also intends to complete development in 1996 of a rapid blood-based antibody test for the Helicobacter pylori bacteria, a pathogen linked to 80% of peptic ulcers and gastric cancers. PMID:12290908

  19. HIV-associated immune complex kidney disease.

    PubMed

    Nobakht, Ehsan; Cohen, Scott D; Rosenberg, Avi Z; Kimmel, Paul L

    2016-05-01

    The introduction in the late 20(th) century of combination antiretroviral therapy (cART) to treat patients infected with HIV has changed the natural history of the disease from an acute illness that rapidly culminates in death, to a chronic condition that can be managed with medications. Over the past decade the epidemiology of kidney disease in US patients infected with HIV has changed, perhaps because of the increased availability and use of cART. Patients with HIV infection exhibit unique immunologic characteristics, including immunodeficiency and dysregulation of immunoglobulin synthetic responses and T-cell function, which can result in glomerular immune complex deposition and subsequent kidney injury. This Review examines the differential diagnoses of HIV-associated immune complex kidney diseases (HIVICD), and discusses the clinical manifestations and mechanisms underlying their development. We address the issues associated with treatment, clinical outcomes, and research needs to enhance our ability to diagnose and optimally treat patients with HIVICD. PMID:26782145

  20. Specific Elimination of Latently HIV-1 Infected Cells Using HIV-1 Protease-Sensitive Toxin Nanocapsules

    PubMed Central

    Wen, Jing; Yan, Ming; Liu, Yang; Li, Jie; Xie, Yiming; Lu, Yunfeng; Kamata, Masakazu; Chen, Irvin S. Y.

    2016-01-01

    Anti-retroviral drugs suppress HIV-1 plasma viremia to undetectable levels; however, latent HIV-1 persists in reservoirs within HIV-1-infected patients. The silent provirus can be activated through the use of drugs, including protein kinase C activators and histone deacetylase inhibitors. This “shock” approach is then followed by “kill” of the producing cells either through direct HIV-1-induced cell death or natural immune mechanisms. However, these mechanisms are relatively slow and effectiveness is unclear. Here, we develop an approach to specifically target and kill cells that are activated early in the process of virus production. We utilize a novel nanocapsule technology whereby the ricin A chain is encapsulated in an inactive form within a polymer shell. Specificity for release of the ricin A toxin is conferred by peptide crosslinkers that are sensitive to cleavage by HIV-1 protease. By using well-established latent infection models, J-Lat and U1 cells, we demonstrate that only within an HIV-1-producing cell expressing functional HIV-1 protease will the nanocapsule release its ricin A cargo, shutting down viral and cellular protein synthesis, and ultimately leading to rapid death of the producer cell. Thus, we provide proof of principle for a novel technology to kill HIV-1-producing cells without effects on non-target cells. PMID:27049645

  1. Specific Elimination of Latently HIV-1 Infected Cells Using HIV-1 Protease-Sensitive Toxin Nanocapsules.

    PubMed

    Wen, Jing; Yan, Ming; Liu, Yang; Li, Jie; Xie, Yiming; Lu, Yunfeng; Kamata, Masakazu; Chen, Irvin S Y

    2016-01-01

    Anti-retroviral drugs suppress HIV-1 plasma viremia to undetectable levels; however, latent HIV-1 persists in reservoirs within HIV-1-infected patients. The silent provirus can be activated through the use of drugs, including protein kinase C activators and histone deacetylase inhibitors. This "shock" approach is then followed by "kill" of the producing cells either through direct HIV-1-induced cell death or natural immune mechanisms. However, these mechanisms are relatively slow and effectiveness is unclear. Here, we develop an approach to specifically target and kill cells that are activated early in the process of virus production. We utilize a novel nanocapsule technology whereby the ricin A chain is encapsulated in an inactive form within a polymer shell. Specificity for release of the ricin A toxin is conferred by peptide crosslinkers that are sensitive to cleavage by HIV-1 protease. By using well-established latent infection models, J-Lat and U1 cells, we demonstrate that only within an HIV-1-producing cell expressing functional HIV-1 protease will the nanocapsule release its ricin A cargo, shutting down viral and cellular protein synthesis, and ultimately leading to rapid death of the producer cell. Thus, we provide proof of principle for a novel technology to kill HIV-1-producing cells without effects on non-target cells. PMID:27049645

  2. Evaluation of a Rapid Point of Care Test for Detecting Acute and Established HIV Infection, and Examining the Role of Study Quality on Diagnostic Accuracy: A Bayesian Meta-Analysis

    PubMed Central

    Smallwood, Megan; Vijh, Rohit; Nauche, Bénédicte; Lebouché, Bertrand; Joseph, Lawrence; Pant Pai, Nitika

    2016-01-01

    Introduction Fourth generation (Ag/Ab combination) point of care HIV tests like the FDA-approved Determine HIV1/2 Ag/Ab Combo test offer the promise of timely detection of acute HIV infection, relevant in the context of HIV control. However, a synthesis of their performance has not yet been done. In this meta-analysis we not only assessed device performance but also evaluated the role of study quality on diagnostic accuracy. Methods Two independent reviewers searched seven databases, including conferences and bibliographies, and independently extracted data from 17 studies. Study quality was assessed with QUADAS-2. Data on sensitivity and specificity (overall, antigen, and antibody) were pooled using a Bayesian hierarchical random effects meta-analysis model. Subgroups were analyzed by blood samples (serum/plasma vs. whole blood) and study designs (case-control vs. cross-sectional). Results The overall specificity of the Determine Combo test was 99.1%, 95% credible interval (CrI) [97.3–99.8]. The overall pooled sensitivity for the device was at 88.5%, 95% [80.1–93.4]. When the components of the test were analyzed separately, the pooled specificities were 99.7%, 95% CrI [96.8–100] and 99.6%, 95% CrI [99.0–99.8], for the antigen and antibody components, respectively. Pooled sensitivity of the antibody component was 97.3%, 95% CrI [60.7–99.9], and pooled sensitivity for the antigen component was found to be 12.3%, 95% (CrI) [1.1–44.2]. No significant differences were found between subgroups by blood sample or study design. However, it was noted that many studies restricted their study sample to p24 antigen or RNA positive specimens, which may have led to underestimation of overall test performance. Detection bias, selection (spectrum) bias, incorporation bias, and verification bias impaired study quality. Conclusions Although the specificity of all test components was high, antigenic sensitivity will merit from an improvement. Besides the accuracy of the

  3. An empiric risk scoring tool for identifying high-risk heterosexual HIV-1 serodiscordant couples for targeted HIV-1 prevention

    PubMed Central

    KAHLE, Erin M.; HUGHES, James P.; LINGAPPA, Jairam R.; JOHN-STEWART, Grace; CELUM, Connie; NAKKU-JOLOBA, Edith; NJUGUNA, Stella; MUGO, Nelly; BUKUSI, Elizabeth; MANONGI, Rachel; BAETEN, Jared M.

    2012-01-01

    Background and objectives Heterosexual HIV-1 serodiscordant couples are increasingly recognized as an important source of new HIV-1 infections in sub-Saharan Africa. A simple risk assessment tool could be useful for identifying couples at highest risk for HIV-1 transmission. Methods Using data from three prospective studies of HIV-1 serodiscordant couples from seven African countries and standard methods for development of clinical prediction rules, we derived and validated a risk scoring tool developed from multivariate modeling and composed of key predictors for HIV-1 risk that could be measured in standard research and clinical settings. Results The final risk score included age of the HIV-1 uninfected partner, married and/or cohabiting partnership, number of children, unprotected sex, uncircumcised male HIV-1 uninfected partner, and plasma HIV-1 RNA in the HIV-1 infected partner. The maximum risk score was 12, scores ≥5 were associated with an annual HIV-1 incidence of >3%, and couples with a score ≥6 accounted for only 28% of the population but 67% of HIV-1 transmissions. The area under the curve for predictive ability of the score was 0.74 (95% CI 0.70–0.78). Internal and external validation showed similar predictive ability of the risk score, even when plasma viral load was excluded from the risk score. Conclusions A discrete combination of clinical and behavioral characteristics defines highest-risk HIV-1 serodiscordant couples. Discriminating highest-risk couples for HIV-1 prevention programs and clinical trials using a validated risk score could improve research efficiency and maximize the impact of prevention strategies for reducing HIV-1 transmission. PMID:23187945

  4. Doctors Report Groundbreaking HIV-To-HIV Organ Transplants

    MedlinePlus

    ... medlineplus/news/fullstory_158039.html Doctors Report Groundbreaking HIV-to-HIV Organ Transplants One donor supplied a kidney to ... News) -- Trailblazing liver and kidney transplants from an HIV-positive donor to HIV-positive recipients were announced ...

  5. Side Effects of HIV Medicines: HIV and Lactic Acidosis

    MedlinePlus

    ... HIV medicines. All HIV medicines in the nucleoside reverse transcriptase inhibitor (NRTI) drug class may cause lactic acidosis, but ... some HIV medicines. HIV medicines in the nucleoside reverse transcriptase inhibitor (NRTI) drug class can cause the body to ...

  6. Intra-Facility Linkage of HIV-Positive Mothers and HIV-Exposed Babies into HIV Chronic Care: Rural and Urban Experience in a Resource Limited Setting

    PubMed Central

    Mugasha, Christine; Kigozi, Joanita; Kiragga, Agnes; Muganzi, Alex; Sewankambo, Nelson; Coutinho, Alex; Nakanjako, Damalie

    2014-01-01

    Introduction Linkage of HIV-infected pregnant women to HIV care remains critical for improvement of maternal and child outcomes through prevention of maternal-to-child transmission of HIV (PMTCT) and subsequent chronic HIV care. This study determined proportions and factors associated with intra-facility linkage to HIV care and Early Infant Diagnosis care (EID) to inform strategic scale up of PMTCT programs. Methods A cross-sectional review of records was done at 2 urban and 3 rural public health care facilities supported by the Infectious Diseases Institute (IDI). HIV-infected pregnant mothers, identified through routine antenatal care (ANC) and HIV-exposed babies were evaluated for enrollment in HIV clinics by 6 weeks post-delivery. Results Overall, 1,025 HIV-infected pregnant mothers were identified during ANC between January and June, 2012; 267/1,025 (26%) in rural and 743/1,025 (74%) in urban facilities. Of these 375/1,025 (37%) were linked to HIV clinics [67/267(25%) rural and 308/758(41%) urban]. Of 636 HIV-exposed babies, 193 (30%) were linked to EID. Linkage of mother-baby pairs to HIV chronic care and EID was 16% (101/636); 8/179 (4.5%)] in rural and 93/457(20.3%) in urban health facilities. Within rural facilities, ANC registration <28 weeks-of-gestation was associated with mothers' linkage to HIV chronic care [AoR, 2.0 95% CI, 1.1–3.7, p = 0.019] and mothers' multi-parity was associated with baby's linkage to EID; AoR 4.4 (1.3–15.1), p = 0.023. Stigma, long distance to health facilities and vertical PMTCT services affected linkage in rural facilities, while peer mothers, infant feeding services, long patient queues and limited privacy hindered linkage to HIV care in urban settings. Conclusion Post-natal linkage of HIV-infected mothers to chronic HIV care and HIV-exposed babies to EID programs was low. Barriers to linkage to HIV care vary in urban and rural settings. We recommend targeted interventions to rapidly improve linkage to

  7. Microdevices for examining immunological responses of single cells to HIV

    PubMed Central

    Choi, Jonghoon; Jeong, Yoon; Han, Hyung-Seop; Lee, Kwan Hyi

    2014-01-01

    More than 60 million people in the world have been diagnosed with HIV infections since the virus was recognized as the causative agent of AIDS in the 1980s. Even though more than half of the infected patients have died, effective disease treatment and prevention measures have not been established. ART (antiretroviral therapy) is the only proven HIV treatment that sustains the suppression of patient viraemia. Current routine approaches to treat HIV infections are targeted at developing vaccines that will induce humoral or cell memory immune responses. However, developing an effective vaccine has been challenging because the HIV mutates rapidly, which allows the virus to evade immune surveillances established against the previous strain. In addition, the virus is able to quickly establish a reservoir and treatment is difficult because of the general lack of knowledge about HIV immune response mechanisms. This review introduces common disease symptoms and the progression of HIV infection with a brief summary of the current treatment approaches. Different cellular immune responses against HIV are also discussed, with emphasis on a nanotechnology research that has focused on probing T-cell response to HIV infection. Furthermore, we discuss recent noteworthy nanotechnology updates on T-cell response screening that is focused on HIV infection. Finally, we review potential future treatment strategies based on the correlations between T-cell response and HIV infection. PMID:25028990

  8. HIV testing in community pharmacies and retail clinics: A model to expand access to screening for HIV infection

    PubMed Central

    Weidle, Paul J.; Lecher, Shirley; Botts, Linda W.; Jones, LaDawna; Spach, David H.; Alvarez, Jorge; Jones, Rhondette; Thomas, Vasavi

    2015-01-01

    Objective To test the feasibility of offering rapid, point-of-care human immunodeficiency virus (HIV) testing at community pharmacies and retail clinics. Design Pilot program to determine how to implement confidential HIV testing services in community pharmacies and retail clinics. Setting 21 community pharmacies and retail clinics serving urban and rural patients in the United States, from August 2011 to July 2013. Participants 106 community pharmacy and retail clinic staff members. Intervention A model was developed to implement confidential HIV counseling and testing services using community pharmacy and retail clinic staff as certified testing providers, or through collaborations with organizations that provide HIV testing. Training materials were developed and sites selected that serve patients from urban and rural areas to pilot test the model. Each site established a relationship with its local health department for HIV testing policies, developed referral lists for confirmatory HIV testing/care, secured a CLIA Certificate of Waiver, and advertised the service. Staff were trained to perform a rapid point-of-care HIV test on oral fluid, and provide patients with confidential test results and information on HIV. Patients with a preliminary positive result were referred to a physician or health department for confirmatory testing and, if needed, HIV clinical care. Main outcome measures Number of HIV tests completed and amount of time required to conduct testing. Results The 21 participating sites administered 1,540 HIV tests, with 1,087 conducted onsite by staff during regular working hours and 453 conducted at 37 different HIV testing events (e.g., local health fairs). The median amount of time required for pretest counseling/consent, waiting for test results, and posttest counseling was 4, 23, and 3 minutes, respectively. A majority of the sites (17) said they planned to continue HIV testing after the project period ended and would seek assistance or support

  9. Rapid diagnosis of Mycobacterium tuberculosis bacteremia by PCR.

    PubMed Central

    Folgueira, L; Delgado, R; Palenque, E; Aguado, J M; Noriega, A R

    1996-01-01

    A method based on DNA amplification and hybridization has been used for the rapid detection of Mycobacterium tuberculosis in blood samples from 38 hospitalized patients (15 human immunodeficiency virus [HIV] positive and 23 HIV negative) in whom localized or disseminated forms of tuberculosis were suspected. In 32 of these patients, the diagnosis of tuberculosis was eventually confirmed by conventional bacteriological or histological procedures. M. tuberculosis DNA was detected with the PCR technique in the peripheral blood mononuclear cells from 9 of 11 (82%) HIV-infected patients and in 7 of 21 (33%) HIV-negative patients (P < 0.01), while M. tuberculosis blood cultures were positive in 1 of 8 (12.5%) and 1 of 18 (5.5%) patients, respectively. PCR was positive in all cases with disseminated disease in both HIV-negative and HIV-positive patients and also in the HIV-positive patients with extrapulmonary tuberculosis. Seven samples from patients with documented illness other than tuberculosis and 12 specimens from healthy volunteers, including seven volunteers with a recent positive purified protein derivative test, were used as controls and had a negative PCR. These results suggest that detection of M. tuberculosis DNA in peripheral blood mononuclear cells may be a useful tool for rapid diagnosis of disseminated and extrapulmonary forms of tuberculosis, especially in an HIV-positive population. PMID:8904404

  10. Use of a High Resolution Melting (HRM) Assay to Compare Gag, Pol, and Env Diversity in Adults with Different Stages of HIV Infection

    PubMed Central

    Cousins, Matthew M.; Laeyendecker, Oliver; Beauchamp, Geetha; Brookmeyer, Ronald; Towler, William I.; Hudelson, Sarah E.; Khaki, Leila; Koblin, Beryl; Chesney, Margaret; Moore, Richard D.; Kelen, Gabor D.; Coates, Thomas; Celum, Connie; Buchbinder, Susan P.; Seage, George R.; Quinn, Thomas C.; Donnell, Deborah; Eshleman, Susan H.

    2011-01-01

    Background Cross-sectional assessment of HIV incidence relies on laboratory methods to discriminate between recent and non-recent HIV infection. Because HIV diversifies over time in infected individuals, HIV diversity may serve as a biomarker for assessing HIV incidence. We used a high resolution melting (HRM) diversity assay to compare HIV diversity in adults with different stages of HIV infection. This assay provides a single numeric HRM score that reflects the level of genetic diversity of HIV in a sample from an infected individual. Methods HIV diversity was measured in 203 adults: 20 with acute HIV infection (RNA positive, antibody negative), 116 with recent HIV infection (tested a median of 189 days after a previous negative HIV test, range 14–540 days), and 67 with non-recent HIV infection (HIV infected >2 years). HRM scores were generated for two regions in gag, one region in pol, and three regions in env. Results Median HRM scores were higher in non-recent infection than in recent infection for all six regions tested. In multivariate models, higher HRM scores in three of the six regions were independently associated with non-recent HIV infection. Conclusions The HRM diversity assay provides a simple, scalable method for measuring HIV diversity. HRM scores, which reflect the genetic diversity in a viral population, may be useful biomarkers for evaluation of HIV incidence, particularly if multiple regions of the HIV genome are examined. PMID:22073290

  11. SIMPLE: An Introduction.

    ERIC Educational Resources Information Center

    Endres, Frank L.

    Symbolic Interactive Matrix Processing Language (SIMPLE) is a conversational matrix-oriented source language suited to a batch or a time-sharing environment. The two modes of operation of SIMPLE are conversational mode and programing mode. This program uses a TAURUS time-sharing system and cathode ray terminals or teletypes. SIMPLE performs all…

  12. HIV Epidemic in Asia: Implications for HIV Vaccine and Other Prevention Trials.

    PubMed

    Phanuphak, Nittaya; Lo, Ying-Ru; Shao, Yiming; Solomon, Sunil Suhas; O'Connell, Robert J; Tovanabutra, Sodsai; Chang, David; Kim, Jerome H; Excler, Jean Louis

    2015-11-01

    An overall decrease of HIV prevalence is now observed in several key Asian countries due to effective prevention programs. The decrease in HIV prevalence and incidence may further improve with the scale-up of combination prevention interventions. The implementation of future prevention trials then faces important challenges. The opportunity to identify heterosexual populations at high risk such as female sex workers may rapidly wane. With unabating HIV epidemics among men who have sex with men (MSM) and transgender (TG) populations, an effective vaccine would likely be the only option to turn the epidemic. It is more likely that efficacy trials will occur among MSM and TG because their higher HIV incidence permits smaller and less costly trials. The constantly evolving patterns of HIV-1 diversity in the region suggest close monitoring of the molecular HIV epidemic in potential target populations for HIV vaccine efficacy trials. CRF01_AE remains predominant in southeast Asian countries and MSM populations in China. This relatively steady pattern is conducive to regional efficacy trials, and as efficacy warrants, to regional licensure. While vaccines inducing nonneutralizing antibodies have promise against HIV acquisition, vaccines designed to induce broadly neutralizing antibodies and cell-mediated immune responses of greater breadth and depth in the mucosal compartments should be considered for testing in MSM and TG. The rationale and design of efficacy trials of combination prevention modalities such as HIV vaccine and preexposure prophylaxis (PrEP) remain hypothetical, require high adherence to PrEP, are more costly, and present new regulatory challenges. The prioritization of prevention interventions should be driven by the HIV epidemic and decided by the country-specific health and regulatory authorities. Modeling the impact and cost-benefit may help this decision process. PMID:26107771

  13. The Oral Bacterial Communities of Children with Well-Controlled HIV Infection and without HIV Infection.

    PubMed

    Goldberg, Brittany E; Mongodin, Emmanuel F; Jones, Cheron E; Chung, Michelle; Fraser, Claire M; Tate, Anupama; Zeichner, Steven L

    2015-01-01

    The oral microbial community (microbiota) plays a critical role in human health and disease. Alterations in the oral microbiota may be associated with disorders such as gingivitis, periodontitis, childhood caries, alveolar osteitis, oral candidiasis and endodontic infections. In the immunosuppressed population, the spectrum of potential oral disease is even broader, encompassing candidiasis, necrotizing gingivitis, parotid gland enlargement, Kaposi's sarcoma, oral warts and other diseases. Here, we used 454 pyrosequencing of bacterial 16S rRNA genes to examine the oral microbiome of saliva, mucosal and tooth samples from HIV-positive and negative children. Patient demographics and clinical characteristics were collected from a cross-section of patients undergoing routine dental care. Multiple specimens from different sampling sites in the mouth were collected for each patient. The goal of the study was to observe the potential diversity of the oral microbiota among individual patients, sample locations, HIV status and various dental characteristics. We found that there were significant differences in the microbiome among the enrolled patients, and between sampling locations. The analysis was complicated by uneven enrollment in the patient cohorts, with only five HIV-negative patients enrolled in the study and by the rapid improvement in the health of HIV-infected children between the time the study was conceived and completed. The generally good oral health of the HIV-negative patients limited the number of dental plaque samples that could be collected. We did not identify significant differences between well-controlled HIV-positive patients and HIV-negative controls, suggesting that well-controlled HIV-positive patients essentially harbor similar oral flora compared to patients without HIV. Nor were significant differences in the oral microbiota identified between different teeth or with different dental characteristics. Additional studies are needed to better

  14. The Oral Bacterial Communities of Children with Well-Controlled HIV Infection and without HIV Infection

    PubMed Central

    Goldberg, Brittany E.; Mongodin, Emmanuel F.; Jones, Cheron E.; Chung, Michelle; Fraser, Claire M.; Tate, Anupama; Zeichner, Steven L.

    2015-01-01

    The oral microbial community (microbiota) plays a critical role in human health and disease. Alterations in the oral microbiota may be associated with disorders such as gingivitis, periodontitis, childhood caries, alveolar osteitis, oral candidiasis and endodontic infections. In the immunosuppressed population, the spectrum of potential oral disease is even broader, encompassing candidiasis, necrotizing gingivitis, parotid gland enlargement, Kaposi’s sarcoma, oral warts and other diseases. Here, we used 454 pyrosequencing of bacterial 16S rRNA genes to examine the oral microbiome of saliva, mucosal and tooth samples from HIV-positive and negative children. Patient demographics and clinical characteristics were collected from a cross-section of patients undergoing routine dental care. Multiple specimens from different sampling sites in the mouth were collected for each patient. The goal of the study was to observe the potential diversity of the oral microbiota among individual patients, sample locations, HIV status and various dental characteristics. We found that there were significant differences in the microbiome among the enrolled patients, and between sampling locations. The analysis was complicated by uneven enrollment in the patient cohorts, with only five HIV-negative patients enrolled in the study and by the rapid improvement in the health of HIV-infected children between the time the study was conceived and completed. The generally good oral health of the HIV-negative patients limited the number of dental plaque samples that could be collected. We did not identify significant differences between well-controlled HIV-positive patients and HIV-negative controls, suggesting that well-controlled HIV-positive patients essentially harbor similar oral flora compared to patients without HIV. Nor were significant differences in the oral microbiota identified between different teeth or with different dental characteristics. Additional studies are needed to better

  15. Maternal HIV status affects the infant hemoglobin level: A comparative cross-sectional study.

    PubMed

    Feleke, Berhanu Elfu

    2016-08-01

    Children, especially infants, are highly vulnerable to iron-deficiency anemia because of their rapid growth of the brain and the rest of the body. The objectives of this study were to compare the prevalence of iron-deficiency anemia in infants born from HIV-positive mothers and HIV-negative mothers and to identify the determinants of iron-deficiency anemia in infants.A comparative cross-sectional study was conducted in Bahir Dar city. Simple random sampling technique was used to select the study participants. Mothers were interviewed; blood samples were collected from mothers and infants to measure the hemoglobin level and anthropometric indicators were obtained from the infants using world health organization standards. Descriptive statistics were used to estimate the prevalence of infantile anemia. Binary logistic regression and multiple linear regressions were used to identify the determinants of infant anemia.A total of 1459 infants born from HIV-positive and HIV-negative mothers were included. The prevalence of iron-deficiency anemia in infants born from HIV-positive and HIV-negative mothers was 41.9% (95% CI: 39-44). Infantile iron-deficiency anemia was associated with maternal HIV infection (adjusted odds ratio [AOR] 2.54 [95% CI: 1.65-3.9]), stunting (AOR 3.46 [95% CI: 2.41-4.97]), low income (AOR 2.72 [95% CI: 2-3.73]), maternal malaria during pregnancy (AOR 1.81 [95% CI: 1.33-2.47]), use of cow milk before 6 month (AOR 1.82 [95% CI: 1.35-2.45]), residence (AOR 0.09 [95% CI: 0.06-0.13]), history of cough or fever 7 days preceding the survey (AOR 2.71 [95% CI: 1.99-3.69]), maternal hemoglobin (B 0.65 [95% CI: 0.61-0.68]), educational status of mother (B 0.22 [95% CI: 0.2-0.23]), age of the mother (B -0.03 [95% CI: -0.03, -0.02]), and family size (B -0.14 [95% CI: -0.18,-0.11]). PMID:27495044

  16. New York State 2010 HIV testing law: an evaluation of testing rates using laboratory data.

    PubMed

    Lazariu, Victoria; Parker, Monica M; Leung, Shu-Yin J; McVoy, Matthew; Gieryic, Susan; Rowe, Kirsten A; Ortega-Peluso, Christina; Anderson, Bridget J; McNutt, Louise-Anne; Smith, Lou C

    2015-01-01

    As of September 2010, New York State (NYS) Public Health Law mandates the offer of HIV testing to all persons aged 13-64 years receiving hospital or primary care services. Changes in the number of HIV tests 13 months before and after law enactment were assessed using HIV test volume data from 166 laboratories holding NYS permits to conduct HIV testing on specimens originating in NYS. Compared with the pre-enactment baseline, overall HIV testing volume increased by 13% following enactment, with the volume of conventional and rapid HIV screening tests increasing by 12.0% and 13.7%, respectively. These data suggest that testing law is having an impact consistent with the legislative intent to increase HIV testing in NYS. Monitoring should be continued to assess testing trends across a variety of health care venues to identify and address additional barriers to HIV testing access. PMID:25545488

  17. Effect of HIV type 1 subtype on virological and immunological response to combination antiretroviral therapy: evidence for a more rapid viral suppression for subtype A than subtype B-infected Greek individuals.

    PubMed

    Paraskevis, Dimirios; Touloumi, Giota; Bakoyannis, Giorgos; Paparizos, Vassilios; Lazanas, Marios; Gargalianos, Panagiotis; Chryssos, Georgios; Antoniadou, Anastasia; Psichogiou, Mina; Panos, Georgios; Katsarou, Olga; Sambatakou, Helen; Kordossis, Theodoros; Hatzakis, Angelos

    2013-03-01

    Whether response to combination antiretroviral therapy (cART) differs between those infected with HIV-1 subtype A or B remains unclear. We compared virological and immunological response to cART in individuals infected with subtype A or B in an ethnically homogeneous population. Data derived from the Athens Multicenter AIDS Cohort Study (AMACS) and analysis were restricted to those of Greek origin. Time to virological response (confirmed HIV-RNA <500 copies/ml) and time to failure (>500 copies/ml at any time or no response by month 6) were analyzed using survival models and CD4 changes after cART initiation using piecewise linear mixed effects models. Of the 571 subjects included in the analysis, 412 (72.2%) were infected with subtype B and 159 (27.8%) with subtype A. After adjusting for various prognostic factors, the rate of virological response was higher for those infected with subtype A versus B (adjusted HR: 1.35; 95% CI: 1.08-1.68; p=0.009). Subtype A was also marginally associated with a lower hazard of virological failure compared to subtype B (HR=0.73; 95% CI: 0.53-1.02; p=0.062). Further adjustment for treatment adherence did not substantially changed the main results. No significant differences were observed in the rates of CD4 increases by subtype. The overall median (95% CI) CD4 increase at 2 years of cART was 193 (175, 212) cells/μl. Our study, based on one of the largest homogeneous groups of subtype A and B infections in Europe, showed that individuals infected with subtype A had an improved virological but similar immunological response to cART compared to those infected with subtype B. PMID:23034083

  18. Evolving T-cell vaccine strategies for HIV, the virus with a thousand faces

    SciTech Connect

    Korber, Bette

    2009-01-01

    HIV's rapid global spread and the human suffering it has left in its wake have made AIDS a global heath priority for the 25 years since its discovery. Yet its capacity to rapidly evolve has made combating this virus a tremendous challenge. The obstacles to creating an effective HIV vaccine are formidable, but there are advances in the field on many fronts, in terms of novel vectors, adjuvants, and antigen design strategies. SIV live attenuated vaccine models are able to confer protection against heterologous challenge, and this continues to provide opportunities to explore the biological underpinnings of a protective effect (9). More indirect, but equally important, is new understanding regarding the biology of acute infection (43), the role of immune response in long-term non-progression (6,62, 81), and defining characteristics of broadly neutralizing antibodies (4). In this review we will focus on summarizing strategies directed towards a single issue, that of contending with HIV variation in terms of designing aT-cell vaccine. The strategies that prove most effective in this area can ultimately be combined with the best strategies under development in other areas, with the hope of ultimately converging on a viable vaccine candidate. Only two large HIV vaccine efficacy trials have been completed and both have failed to prevent infection or confer a benefit to infected individual (23,34), but there is ample reason to continue our efforts. A historic breakthrough came in 1996, when it was realized that although the virus could escape from a single antiretroviral (ARV) therapy, it could be thwarted by a combination of medications that simultaneously targeted different parts of the virus (HAART) (38). This revelation came after 15 years of research, thought, and clinical testing; to enable that vital progress the research and clinical communities had to first define and understand, then develop a strategy to counter, the remarkable evolutionary potential of the virus

  19. International epidemiology of HIV and AIDS among injecting drug users.

    PubMed

    Des Jarlais, D C; Friedman, S R; Choopanya, K; Vanichseni, S; Ward, T P

    1992-10-01

    HIV/AIDS and iv drug use (IVDU) are of significant multinational scope and growing. Supporting increased IVDU in many countries are countries' geographical proximity to illicit drug trafficking distribution routes, law enforcement efforts which increase the demand for more efficient drug distribution and consumption, and countries' infrastructural and social modernization. Given the failures of intensified law enforcement efforts to thwart the use and proliferation of illegal drugs, countries with substantial IVDU should look away from preventing use to preventing HIV transmission within drug user populations. With HIV seroprevalence rates rapidly reaching 40-50% in some developing country IVDU groups, a variety of prevention programs is warranted. Such programs should be supported and implemented while prevention remains feasible. This paper examines the variation in HIV seroprevalence among IVD users, rapid HIV spread among users, HIV among IVDUs in Bangkok, emerging issues in HIV transmission among IVDUs, non-AIDS manifestations of HIV infection among IVDUs, prevention programs and effectiveness, and harm reduction. PMID:1466837

  20. HIV knowledge, stigma, and illness beliefs among pediatric caregivers in Ghana who have not disclosed their child's HIV status

    PubMed Central

    Paintsil, Elijah; Renner, Lorna; Antwi, Sampson; Dame, Joycelyn; Enimil, Anthony; Ofori-Atta, Angela; Alhassan, Amina; Ofori, Irene Pokuaa; Cong, Xiangyu; Kyriakides, Tassos; Reynolds, Nancy R.

    2015-01-01

    The majority of HIV-infected children in sub-Saharan Africa have not been informed of their HIV status. Caregivers are reluctant to disclose HIV status to their children because of concern about the child’s ability to understand, parental sense of guilt, and fear of social rejection and isolation. We hypothesized that the low prevalence of pediatric HIV disclosure in Ghana is due to lack of accurate HIV information and high HIV stigma among caregivers. This is a preliminary analysis of baseline data of an HIV pediatric disclosure intervention study in Ghana (“Sankofa”). “Sankofa” – is a two-arm randomized controlled clinical trial comparing disclosure intervention plus usual care (intervention arm) vs usual care (control arm) at Korle-Bu Teaching Hospital (KBTH; control arm) and Komfo-Anokye Teaching Hospital (KATH; intervention arm). We enrolled HIV-infected children, ages 7–18 years who do not know their HIV status, and their caregivers. Baseline data of caregivers included demographic characteristics; Brief HIV Knowledge Questionnaire (HIV-KQ-18); Brief Illness Perception Questionnaire; and HIV Stigma Scale. Simple and multivariable linear regression analyses were used to assess the relationship between caregiver characteristics and HIV knowledge, stigma, and illness perception. Two hundred and ninety-eight caregivers were enrolled between January 2013 and July 2014 at the two study sites; KBTH (n = 167) and KATH (n = 131). The median age of caregivers was 41 years; 80.5% of them were female and about 60% of caregivers were HIV-positive. Seventy-eight percent of caregivers were self-employed with low household income. In both unadjusted and adjusted analyses, HIV negative status and lower level of education were associated with poor scores on HIV-KQ. HIV positive status remained significant for higher level of stigma in the adjusted analyses. None of the caregiver’s characteristics predicted caregiver’s illness perception. Intensification of

  1. Adolescent HIV/AIDS: Issues and challenges.

    PubMed

    Naswa, Smriti; Marfatia, Y S

    2010-01-01

    Adolescence (10-19 years) is a phase of physical growth and development accompanied by sexual maturation, often leading to intimate relationships. Adolescent HIV/AIDS is a separate epidemic and needs to be handled and managed separately from adult HIV. The adolescents can be subdivided into student, slum and street youth; street adolescents being most vulnerable to HIV/AIDS. Among various risk factors and situations for adolescents contracting HIV virus are adolescent sex workers, child trafficking, child labor, migrant population, childhood sexual abuse, coercive sex with an older person and biologic (immature reproductive tract) as well as psychological vulnerability. The most common mode of transmission is heterosexual, yet increasing number of perinatally infected children are entering adolescence. This is due to "bimodal progression" (rapid and slow progressors) among the vertically infected children. Clinically, the HIV infected adolescents present as physically stunted individuals, with delayed puberty and adrenarche. Mental illness and substance abuse are important co-morbidities. The disclosure and declaration of HIV status to self and family is challenging and guilt in sexually infected adolescents and tendency to blame parents if vertically affected need special consideration and proper counseling. Serodiscordance of the twins and difference in disease progression of seroconcordant twins are added causes of emotional trauma. Treatment related issues revolve around the when and what of initiation of ART; the choice of antiretrovirals and their dosages; issues related to long term ADRs; sense of disinhibition following ART commencement; adherence and resistance. PMID:21808429

  2. Adolescent HIV treatment issues in South Africa.

    PubMed

    Dawood, H

    2015-11-01

    Following the discovery of the human immunodeficiency virus (HIV), our knowledge of HIV infection and management has increased rapidly, but implementation of interventions has been slow in resource-limited settings. In particular, interventions such as antiretroviral treatment (ART) and prevention of mother-to-child transmission were hindered owing to lack of access to antiretroviral drugs. This resulted in ongoing HIV transmission, morbidity and mortality associated with opportunistic infections. Notwithstanding the current progress in HIV prevention and treatment, challenges remain in preventing new infections in adolescents and supporting and treating HIV-infected adolescents. Barriers to successful treatment of infection in adolescents include denial of diagnosis, poor understanding or perception of future benefits of treatment and current-orientated thinking that may contribute to non-adherence to ART. Side-effects that lead to stigmatisation, such as lipoatrophy (stavudine, zidovudine), diarrhoea and flatulence (lopinavir/ritonavir) and gynaecomastia (efavirenz), maybe intolerable and prevent adherence to treatment. This article highlights common treatment issues in HIV adolescent care and provides guidance on their management in the South African setting. PMID:26937511

  3. HIV Risk Perception and Behavior among Sex Workers in Three Major Urban Centers of Mozambique

    PubMed Central

    Langa, Judite; Sousa, César; Sidat, Mohsin; Kroeger, Karen; McLellan-Lemal, Eleanor; Belani, Hrishikesh; Patel, Shama; Shodell, Daniel; Shodell, Michael; Benech, Irene; Needle, Richard

    2014-01-01

    HIV risk perceptions and behaviors of 236 commercial sex workers from three major Mozambican urban centers were studied using the International Rapid Assessment, Response and Evaluation (I-RARE) methodology. All were offered HIV testing and, in Maputo, syphilis testing was offered as well. Sixty-three of the 236 opted for HIV testing, with 30 (48%) testing positive for HIV. In Maputo, all 30 receiving HIV tests also had syphilis testing, with 6 (20%) found to be positive. Results include interview excerpts and qualitative results using I-RARE methodology and AnSWR-assisted analyses of the interviews and focus group sessions. PMID:24736653

  4. Potent Cell-Intrinsic Immune Responses in Dendritic Cells Facilitate HIV-1-Specific T Cell Immunity in HIV-1 Elite Controllers

    PubMed Central

    Martin-Gayo, Enrique; Buzon, Maria Jose; Ouyang, Zhengyu; Hickman, Taylor; Cronin, Jacqueline; Pimenova, Dina; Walker, Bruce D.; Lichterfeld, Mathias; Yu, Xu G.

    2015-01-01

    The majority of HIV-1 elite controllers (EC) restrict HIV-1 replication through highly functional HIV-1-specific T cell responses, but mechanisms supporting the evolution of effective HIV-1-specific T cell immunity in these patients remain undefined. Cytosolic immune recognition of HIV-1 in conventional dendritic cells (cDC) can facilitate priming and expansion of HIV-1-specific T cells; however, HIV-1 seems to be able to avoid intracellular immune recognition in cDCs in most infected individuals. Here, we show that exposure of cDCs from EC to HIV-1 leads to a rapid and sustained production of type I interferons and upregulation of several interferon-stimulated effector genes. Emergence of these cell-intrinsic immune responses was associated with a reduced induction of SAMHD1 and LEDGF/p75, and an accumulation of viral reverse transcripts, but inhibited by pharmacological blockade of viral reverse transcription or siRNA-mediated silencing of the cytosolic DNA sensor cGAS. Importantly, improved cell-intrinsic immune recognition of HIV-1 in cDCs from elite controllers translated into stronger abilities to stimulate and expand HIV-1-specific CD8 T cell responses. These data suggest an important role of cell-intrinsic type I interferon secretion in dendritic cells for the induction of effective HIV-1-specific CD8 T cells, and may be helpful for eliciting functional T cell immunity against HIV-1 for preventative or therapeutic clinical purposes. PMID:26067651

  5. The rapid evolution of complex systems from simple beginnings

    NASA Technical Reports Server (NTRS)

    Fox, S. W.

    1971-01-01

    Methods for the geological production of proteinous amino acids are examined. Particular attention was given to organic evolution and thermal polymerization. Also included are data on amino acid reactions after polymerization.

  6. Behavioural risk factors for HIV/AIDS in a low-HIV prevalence Muslim nation: Bangladesh.

    PubMed

    Gibney, L; Choudhury, P; Khawaja, Z; Sarker, M; Vermund, S H

    1999-03-01

    A review of published and unpublished data indicates the prevalence of high-risk behaviours for HIV transmission in segments of the Bangladeshi population. These include casual unprotected sex, heterosexual as well as between males, prior to and after marriage. Intravenous drug use (IVDU) exists though illicit drugs are more commonly inhaled. There is a fear, however, that inhalers may turn to injecting drugs, as is common in neighbouring countries. The lack of public awareness of HIV/AIDS, and misconceptions about the disease, may contribute to continued high-risk behaviours by segments of the population and, thus, to the spread of HIV. Bangladesh's proximity to India and Myanmar (countries with high HIV endemicity and a rapidly growing number of cases) increases fears of an epidemic in Bangladesh. This proximity will only be a risk factor, however, if high-risk contacts occur between nationals of these countries. PMID:10340200

  7. Behavioural risk factors for HIV/AIDS in a low-HIV prevalence Muslim nation: Bangladesh

    PubMed Central

    Gibney, L; Choudhury, P; Khawaja, Z; Sarker, M; Vermund, SH

    2008-01-01

    Summary A review of published and unpublished data indicates the prevalence of high-risk behaviours for HIV transmission in segments of the Bangladeshi population. These include casual unprotected sex, heterosexual as well as between males, prior to and after marriage. Intravenous drug use (IVDU) exists though illicit drugs are more commonly inhaled. There is a fear, however, that inhalers may turn to injecting drugs, as is common in neighbouring countries. The lack of public awareness of HIV/AIDS, and misconceptions about the disease, may contribute to continued high-risk behaviours by segments of the population and, thus, to the spread of HIV. Bangladesh’s proximity to India and Myanmar (countries with high HIV endemicity and a rapidly growing number of cases) increases fears of an epidemic in Bangladesh. This proximity will only be a risk factor, however, if high-risk contacts occur between nationals of these countries. PMID:10340200

  8. Polyclonal B-cell activation reveals antibodies against human immunodeficiency virus type 1 (HIV-1) in HIV-1-seronegative individuals.

    PubMed Central

    Jehuda-Cohen, T; Slade, B A; Powell, J D; Villinger, F; De, B; Folks, T M; McClure, H M; Sell, K W; Ahmed-Ansari, A

    1990-01-01

    Identification of human immunodeficiency virus type 1 (HIV-1)-infected individuals is of paramount importance for the control of the spread of AIDS worldwide. Currently, the vast majority of screening centers throughout the world rely on serological techniques. As such, clinically asymptomatic but HIV-infected, seronegative individuals are rarely identified. In this report we show that 18% (30/165) of seronegative individuals who were considered to be a unique cohort of patients at high risk for HIV infection had circulating B cells that, upon in vitro polyclonal activation with pokeweed mitogen, produced antibodies reactive with HIV. Furthermore, polymerase chain reaction analysis of DNA obtained from aliquots of the peripheral blood mononuclear cells from these seronegative but pokeweed mitogen assay-positive individuals tested revealed the presence of HIV-specific sequences in a significant number of samples. In addition, depletion of CD8+ T cells from peripheral blood mononuclear cells of HIV-1-seronegative individuals prior to in vitro culture with pokeweed mitogen resulted in increased sensitivity for detecting HIV-reactive antibodies. This assay has obvious epidemiological implications, especially in the case of high-risk groups, and also provides a simple technique to enhance detection of HIV-infected individuals. Of further interest is the determination of the mechanisms related to the lack of HIV-specific antibodies in the serum of these infected individuals. Images PMID:2111024

  9. Genome editing strategies: potential tools for eradicating HIV-1/AIDS

    PubMed Central

    Khalili, Kamel; Gordon, Jennifer; Cosentino, Laura; Hu, Wenhui

    2015-01-01

    Current therapy for controlling HIV-1 infection and preventing AIDS progression has profoundly decreased viral replication in cells susceptible to HIV-1 infection, but it does not eliminate the low level of viral replication in latently infected cells which contain integrated copies of HIV-1 proviral DNA. There is an urgent need for the development of HIV-1 genome eradication strategies that will lead to a permanent or “sterile” cure of HIV-1/AIDS. In the past few years, novel nuclease-initiated genome editing tools have been developing rapidly, including ZFNs, TALENs, and the CRISPR/Cas9 system. These surgical knives, which can excise any genome, provide a great opportunity to eradicate the HIV-1 genome by targeting highly conserved regions of the HIV-1 long terminal repeats or essential viral genes. Given the time consuming and costly engineering of target-specific ZFNs and TALENs, the RNA-guided endonuclease Cas9 technology has emerged as a simpler and more versatile technology to allow permanent removal of integrated HIV-1 proviral DNA in eukaryotic cells, and hopefully animal models or human patients. The major unmet challenges of this approach at present include inefficient nuclease gene delivery, potential off-target cleavage, and cell-specific genome targeting. Nanoparticle or lentivirus-mediated delivery of next generation Cas9 technologies including nickase or RNA-guided FokI nuclease (RFN) will further improve the potential for genome editing to become a promising approach for curing HIV-1/AIDS. PMID:25716921

  10. Testing for HIV

    MedlinePlus

    ... Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Vaccines, Blood & Biologics Home Vaccines, Blood & Biologics Safety & Availability (Biologics) HIV Home Test Kits Testing for HIV Share Tweet Linkedin Pin it More ...

  11. HIV/AIDS

    MedlinePlus

    ... immunodeficiency virus (HIV) is the virus that causes AIDS. When a person becomes infected with HIV, the ... cancers. When that happens, the illness is called AIDS. Once a person has the virus, it stays ...

  12. HIV and AIDS

    MedlinePlus

    ... that causes the disease AIDS. HIV Hurts the Immune System People who are HIV positive have been tested ... to everyone in the world. When the person's immune system has weakened and more of the blood's T ...

  13. Microbiome in HIV infection

    PubMed Central

    Salas, January T.; Chang, Theresa L.

    2014-01-01

    HIV primary infection occurs at mucosa tissues, suggesting an intricate interplay between microbiome and HIV infection. Recent advanced technologies of high-throughput sequencing and bioinformatics allow researchers to explore nonculturable microbes including bacteria, virus and fungi and their association with diseases. HIV/SIV infection is associated with microbiome shifts and immune activation that may affect the outcome of disease progression. Similarly, altered microbiome and inflammation are associated with increased risks of HIV acquisition, suggesting the role of microbiome in HIV transmission. In this review, we will focus on microbiome in HIV infection at various mucosal compartments. Understanding the relationship between microbiome and HIV may offer insights into development of better strategies for HIV prevention and treatment. PMID:25439273

  14. HIV and Cardiovascular Disease

    MedlinePlus

    ... CVD. ART can increase blood fats (cholesterol and triglycerides, see fact sheet 123.) It can also help ... disease. HIV infection decreases good cholesterol and increases triglycerides. HIV causes inflammation. This can also contribute to ...

  15. Older People and HIV

    MedlinePlus

    ... Many older people believe that HIV only affects younger people Most older people get little training in ... diseases among older people, as they do for younger people. Physicians may not diagnose HIV infection in ...

  16. Smoking and HIV

    MedlinePlus

    ... 28, 2014 Select a Language: Fact Sheet 803 Smoking and HIV WHY IS SMOKING MORE DANGEROUS FOR ... It can also worsen liver problems like hepatitis. Smoking and Side Effects People with HIV who smoke ...

  17. HIV and Pregnancy

    MedlinePlus

    ... Pregnancy Patient Education FAQs HIV and Pregnancy Patient Education Pamphlets - Spanish HIV and Pregnancy FAQ113, December 2012 PDF Format ... Your Practice Patient Safety & Quality Payment Reform (MACRA) Education & Events Annual ... Pamphlets Teen Health About ACOG About Us Leadership & ...

  18. Total HIV-1 DNA, a Marker of Viral Reservoir Dynamics with Clinical Implications.

    PubMed

    Avettand-Fènoël, Véronique; Hocqueloux, Laurent; Ghosn, Jade; Cheret, Antoine; Frange, Pierre; Melard, Adeline; Viard, Jean-Paul; Rouzioux, Christine

    2016-10-01

    HIV-1 DNA persists in infected cells despite combined antiretroviral therapy (cART), forming viral reservoirs. Recent trials of strategies targeting latent HIV reservoirs have rekindled hopes of curing HIV infection, and reliable markers are thus needed to evaluate viral reservoirs. Total HIV DNA quantification is simple, standardized, sensitive, and reproducible. Total HIV DNA load influences the course of the infection and is therefore clinically relevant. In particular, it is predictive of progression to AIDS and death, independently of HIV RNA load and the CD4 cell count. Baseline total HIV DNA load is predictive of the response to cART. It declines during cART but remains quantifiable, at a level that reflects both the history of infection (HIV RNA zenith, CD4 cell count nadir) and treatment efficacy (residual viremia, cumulative viremia, immune restoration, immune cell activation). Total HIV DNA load in blood is also predictive of the presence and severity of some HIV-1-associated end-organ disorders. It can be useful to guide individual treatment, notably, therapeutic de-escalation. Although it does not distinguish between replication-competent and -defective latent viruses, the total HIV DNA load in blood, tissues, and cells provides insights into HIV pathogenesis, probably because all viral forms participate in host cell activation and HIV pathogenesis. Total HIV DNA is thus a biomarker of HIV reservoirs, which can be defined as all infected cells and tissues containing all forms of HIV persistence that participate in pathogenesis. This participation may occur through the production of new virions, creating new cycles of infection and disseminating infected cells; maintenance or amplification of reservoirs by homeostatic cell proliferation; and viral transcription and synthesis of viral proteins without new virion production. These proteins can induce immune activation, thus participating in the vicious circle of HIV pathogenesis. PMID:27559075

  19. Immunogenetics of HIV and HIV associated tuberculosis.

    PubMed

    Raghavan, S; Alagarasu, K; Selvaraj, P

    2012-01-01

    Tuberculosis (TB) is the frequent major opportunistic infection in HIV-infected patients, and is the leading cause of mortality among HIV-infected patients. Genetic susceptibility to TB in HIV negative subjects is well documented. Since coinfections can influence the way in which immune system respond to different pathogens, genetic susceptibility to TB in HIV patients might also change. Studies from India and other parts of the world have shown that genetic susceptibility to TB is influenced by HIV infection. In the present review, we emphasize the role of genetic factors in determining susceptibility to HIV infection, disease progression and development of TB in HIV-infected patients. Polymorphisms in human leukocyte antigen (HLA), MBL2, CD209, vitamin D receptor, cytokine, chemokine and chemokine receptor genes have been shown to be associated with development of TB in HIV patients. However, the results are inconclusive and larger well-defined studies with precise clinical data are required to validate these associations. Apart from candidate gene approach, genome-wide association studies are also needed to unravel the unknown or to establish the previously reported genetic associations with HIV associated TB. Despite the preliminary status of the reported associations, it is becoming clear that susceptibility to development of TB in HIV patients is influenced by both environmental and genetic components. Understanding the genetic and immunologic factors that influence susceptibility to TB in HIV patients could lead to novel insights for vaccine development as well as diagnostic advances to target treatment to those who are at risk for developing active disease. PMID:21943869

  20. The effect of delayed death in HIV/AIDS models.

    PubMed

    Lutambi, Angelina Mageni

    2016-01-01

    HIV-infected patients who receive treatment survive for some years after they have acquired the disease. The received treatment causes sustained reduction of viral reproduction by improving the immune function, leading to prolonged progression period to AIDS development. This prolonged progression period has created variability in survival times that affects estimates produced using mathematical models that do not include delay in disease related mortality. This paper investigates the effect of including delay in AIDS death occurrence in HIV/AIDS transmission models. A simple mathematical model with two stages of HIV progression is developed and extended to include time delay in the occurrence of AIDS deaths. Numerical simulations indicate that time delay changes the mortality curves considerably but has less effect on the proportion of infectives. The study highlights the importance of incorporating delay in models of HIV/AIDS for the production of accurate HIV/AIDS estimates. PMID:27141921

  1. Reduce HIV Risk

    MedlinePlus

    ... are increasing among younger people from 13 to 30 years of age. The key to defeating HIV lies ... Control and Prevention (CDC) has used them as models, and Dr. Jemmott was invited to South Africa to help decrease HIV/AIDS there. "For the past 15 years, I have observed how the HIV/AIDS epidemic ...

  2. HIV and the menopause.

    PubMed

    Fan, Maria D; Maslow, Bat-Sheva; Santoro, Nanette; Schoenbaum, Ellie

    2008-12-01

    Dramatic improvement in the survival of the HIV population has occurred with the ascendance of highly active antiretroviral therapy (HAART). In the foreseeable future, HIV-infected women who acquired disease during the peak years of the epidemic are expected to survive to experience menopause and even years beyond. The HIV epidemic may be viewed as 'mature', as its earlier victims become part of the geriatric population. Research about the process of menopause in HIV-infected women and, conversely, about HIV infection in women undergoing menopause is currently limited. Existing research suggests that the process of menopause is affected by HIV infection, inasmuch as infected women appear to experience menopause at an earlier age, with greater symptomatology, and with different reproductive hormone profiles compared with HIV-uninfected women. HIV infection also appears to affect bone mineral density, cardiovascular disease and cognition, with some age-related interactions. Lifestyle and demographic factors have pervasive importance for both HIV infection and the menopause in women. This article reviews the current state of knowledge about the menopausal process in HIV-infected women, and the common conditions in postmenopausal women that are likely to be affected by HIV infection. Clinicians should appreciate the potential role of HIV infection in caring for menopause-aged women. PMID:19037065

  3. HIV Disease: Current Concepts.

    ERIC Educational Resources Information Center

    Keeling, Richard P.

    1993-01-01

    Describes human immunodeficiency virus (HIV), newly characterized human retrovirus which causes chronic, progressive, immune deficiency disease, the most severe phase of which is Acquired Immune Deficiency Syndrome (AIDS). Reviews most important current epidemiologic, clinical, and virologic information about HIV and HIV disease and provides…

  4. Simple pulmonary eosinophilia

    MedlinePlus

    Pulmonary infiltrates with eosinophilia; Loffler syndrome; Eosinophilic pneumonia; Pneumonia - eosinophilic ... simple pulmonary eosinophilia is a severe type of pneumonia called acute idiopathic eosinophilic pneumonia.

  5. HIV Acquisition Among Women From Selected Areas of the United States

    PubMed Central

    Hodder, Sally L.; Justman, Jessica; Hughes, James P.; Wang, Jing; Haley, Danielle F.; Adimora, Adaora A.; Del Rio, Carlos; Golin, Carol E.; Kuo, Irene; Rompalo, Anne; Soto-Torres, Lydia; Mannheimer, Sharon B.; Johnson-Lewis, LeTanya; Eshleman, Susan H.; El-Sadr, Wafaa M.

    2014-01-01

    Background Women account for 23% of newly diagnosed HIV infections in the United States, but there are few recent, well-characterized cohorts of U.S. women in whom behavior characteristics and HIV acquisition have been well-described. Objective To evaluate HIV incidence and describe behaviors among U.S. women residing in areas of high HIV prevalence. Design Multisite, longitudinal cohort of women who had HIV rapid testing and audio computer-assisted self-interviews at baseline and every 6 months for up to 12 months. (ClinicalTrials.gov: NCT00995176) Setting 10 urban and periurban communities with high HIV prevalence and poverty rates, located in the northeastern and southeastern United States. Patients Venue-based sampling was used to recruit women aged 18 to 44 years who recently had unprotected sex and had 1 or more additional personal or partner risk factors and no self-reported previous HIV diagnosis. Measurements HIV prevalence and incidence, frequency of HIV risk behaviors, and health status perceptions. Results Among 2099 high-risk women (85.9% black and 11.7% of Hispanic ethnicity), 32 (1.5%) were diagnosed with HIV infection at enrollment. Annual HIV incidence was 0.32% (95% CI, 0.14% to 0.74%). Older age, substance use, and knowing a partner had HIV were associated with HIV prevalence. Ten women died during the study (0.61% per year). Limitations Longitudinal assessment of risk behaviors was limited to a maximum of 12 months. There were few incident HIV infections, precluding identification of characteristics predictive of HIV acquisition. Conclusion This study enrolled a cohort of women with HIV incidence substantially higher than the Centers for Disease Control and Prevention national estimate in the general population of U.S. black women. Concerted efforts to improve preventive health care strategies for HIV and overall health status are needed for similar populations. PMID:23277896

  6. Impact of HLA-driven HIV adaptation on virulence in populations of high HIV seroprevalence

    PubMed Central

    Payne, Rebecca; Muenchhoff, Maximilian; Mann, Jaclyn; Roberts, Hannah E.; Matthews, Philippa; Adland, Emily; Hempenstall, Allison; Huang, Kuan-Hsiang; Brockman, Mark; Brumme, Zabrina; Sinclair, Marc; Miura, Toshiyuki; Frater, John; Essex, Myron; Shapiro, Roger; Walker, Bruce D.; Ndung’u, Thumbi; McLean, Angela R.; Carlson, Jonathan M.; Goulder, Philip J. R.

    2014-01-01

    It is widely believed that epidemics in new hosts diminish in virulence over time, with natural selection favoring pathogens that cause minimal disease. However, a tradeoff frequently exists between high virulence shortening host survival on the one hand but allowing faster transmission on the other. This is the case in HIV infection, where high viral loads increase transmission risk per coital act but reduce host longevity. We here investigate the impact on HIV virulence of HIV adaptation to HLA molecules that protect against disease progression, such as HLA-B*57 and HLA-B*58:01. We analyzed cohorts in Botswana and South Africa, two countries severely affected by the HIV epidemic. In Botswana, where the epidemic started earlier and adult seroprevalence has been higher, HIV adaptation to HLA including HLA-B*57/58:01 is greater compared with South Africa (P = 7 × 10−82), the protective effect of HLA-B*57/58:01 is absent (P = 0.0002), and population viral replicative capacity is lower (P = 0.03). These data suggest that viral evolution is occurring relatively rapidly, and that adaptation of HIV to the most protective HLA alleles may contribute to a lowering of viral replication capacity at the population level, and a consequent reduction in HIV virulence over time. The potential role in this process played by increasing antiretroviral therapy (ART) access is also explored. Models developed here suggest distinct benefits of ART, in addition to reducing HIV disease and transmission, in driving declines in HIV virulence over the course of the epidemic, thereby accelerating the effects of HLA-mediated viral adaptation. PMID:25453107

  7. HIV multidisciplinary teams work: support services improve access to and retention in HIV primary care.

    PubMed

    Sherer, R; Stieglitz, K; Narra, J; Jasek, J; Green, L; Moore, B; Shott, S; Cohen, M

    2002-08-01

    The multidisciplinary team model of HIV care evolved out of necessity due to the diverse characteristics and needs of people living with HIV disease. Though it is now accepted as the international standard of care, it represents a significant departure from methods of care for other infectious diseases, and debate continues regarding the effectiveness of its interventions. The debate has been largely uninformed by data; for example, little is known about the relationship between ancillary support services and primary care outcomes. We hypothesized that support services increase access to and retention in HIV primary care in an inner city public hospital clinic. We conducted a retrospective analysis of clinical data sets on 2,647 patients at the CORE Center, Chicago from 1997-1998 to investigate the relationship between four support services-case management (CM), transportation (TRANS), mental health (MH) and chemical dependency (CD)-and access to and retention in HIV primary care. We found that patients who received each of these services were significantly more likely to receive any care, regular care and had more visits than patients with no service, and retention increased by 15-18%. Female gender, younger age, self-pay status and IDU predicted less regular care. Need for all services was substantial and significantly greater in women. Outcomes improved to the greatest extent among patients who needed and received each service. We conclude that support services significantly increased access to and retention in HIV primary care. Our findings validate the multidisciplinary team model of HIV care, and suggest that health services that are tailored to the express needs of patients lead to better care and improved health outcomes. Further testing of changes in health care delivery to meet the rapidly changing needs of people living with HIV disease and respond to the constantly changing practice of HIV medicine is urgently needed to maintain and extend the advances

  8. Hospital treatment of HIV patients.

    PubMed

    Ola, Samuel Olawale

    2006-12-01

    Treatment of patients with HIV/AIDS in Nigeria has progressed from the stage of inactivity, unconcern, abandonment and neglect to the present stage of holistic care involving treatment of the infection with Highly Active Anti Retroviral Agents, complications of the disease and side effects of antiretroviral therapy as well as that of human behavioural responses towards the disease with hope and promising outcome. The goal of the treatment is to prolong the patient's life while maintaining the best possible quality of health and life. It is now a continuum of care between the hospital and the different sectors of the community. Hospital treatment of patients with HIV-AIDS is complex and yet a simple task if there is healthy interaction of the patients and health care providers in a milieu of well equipped hospital setting with available treatment facilities for proper management of diseases. Similarly, for the care to achieve its goal, it requires a joint participation of the community and the commitment of the government not only on curtailment of the reservoir of HIV infection by antiretroviral therapy but total eradication of diseases, poverty and ignorance in all its entirety. PMID:18050774

  9. Characterization of a folding intermediate from HIV-1 ribonuclease H.

    PubMed Central

    Kern, G.; Handel, T.; Marqusee, S.

    1998-01-01

    The RNase H domain from HIV-1 (HIV RNase H) encodes an essential retroviral activity. Refolding of the isolated HIV RNase H domain shows a kinetic intermediate detectable by stopped-flow far UV circular dichroism and pulse-labeling H/D exchange. In this intermediate, strands 1, 4, and 5 as well as helices A and D appear to be structured. Compared to its homolog from Escherichia coli, the rate limiting step in refolding of HIV RNase H appears closer to the native state. We have modeled this kinetic intermediate using a C-terminal deletion fragment lacking helix E. Like the kinetic intermediate, this variant folds rapidly and shows a decrease in stability. We propose that inhibition of the docking of helix E to this folding intermediate may present a novel strategy for anti HIV-1 therapy. PMID:9792104

  10. HIV/AIDS knowledge and self-esteem among adolescents.

    PubMed

    Oxley, G M

    2001-05-01

    The incidence of HIV/AIDS is rapidly increasing among adolescents and young adults with some studies linking sexual risk taking and self-esteem. A convenience sample of 39 ethnically diverse adolescents, ages 14-18, participated in a pilot study designed to assess HIV/AIDS knowledge and to build self-esteem. Adolescents selected from two centers in California completed the Coopersmith Self-Esteem Inventory and the Student Health Questionnaire (SHQ) before beginning and after completing a program of six 2-hour educational sessions. These sessions focused on HIV/AIDS knowledge and building self-esteem. Knowledge of HIV/AIDS prevention and transmission increased by 2096 from pretest to posttest. Practitioners addressing the needs of adolescents should focus on in-depth information regarding HIV/AIDS, especially in the area of prevention strategies and cultural factors influencing levels of self-esteem. PMID:11881719

  11. Rapid Prototyping

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Javelin, a Lone Peak Engineering Inc. Company has introduced the SteamRoller(TM) System as a commercial product. The system was designed by Javelin during a Phase II NASA funded small commercial product. The purpose of the invention was to allow automated-feed of flexible ceramic tapes to the Laminated Object Manufacturing rapid prototyping equipment. The ceramic material that Javelin was working with during the Phase II project is silicon nitride. This engineered ceramic material is of interest for space-based component.

  12. Laparoscopic simple prostatectomy.

    PubMed

    Blew, Brian D M; Fazio, Luke M; Pace, Kenneth; D'A Honey, R John

    2005-12-01

    Classically, surgical options for very large prostate glands, not amenable to transurethral resection, include suprapubic or retropubic simple prostatectomy and Holmium laser enucleation of the prostate (HoLEP). We pres