Browsing by Subject "drug screening"

Now showing 1 - 5 of 5
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    A cell-based high-throughput screening method to directly examine transthyretin amyloid fibril formation at neutral pH
    (Elsevier, 2019-07-19) Ueda, Mitsuharu; Okada, Masamitsu; Mizuguchi, Mineyuki; Kluve-Beckerman, Barbara; Kanenawa, Kyosuke; Isoguchi, Aito; Misumi, Yohei; Tasaki, Masayoshi; Ueda, Akihiko; Kanai, Akinori; Sasaki, Ryoko; Masuda, Teruaki; Inoue, Yasuteru; Nomura, Toshiya; Shinriki, Satoru; Shuto, Tsuyoshi; Kai, Hirofumi; Yamashita, Taro; Matsui, Hirotaka; Benson, Merrill D.; Ando, Yukio; Pathology and Laboratory Medicine, School of Medicine
    Transthyretin (TTR) is a major amyloidogenic protein associated with hereditary (ATTRm) and nonhereditary (ATTRwt) intractable systemic transthyretin amyloidosis. The pathological mechanisms of ATTR-associated amyloid fibril formation are incompletely understood, and there is a need for identifying compounds that target ATTR. C-terminal TTR fragments are often present in amyloid-laden tissues of most patients with ATTR amyloidosis, and on the basis of in vitro studies, these fragments have been proposed to play important roles in amyloid formation. Here, we found that experimentally-formed aggregates of full-length TTR are cleaved into C-terminal fragments, which were also identified in patients' amyloid-laden tissues and in SH-SY5Y neuronal and U87MG glial cells. We observed that a 5-kDa C-terminal fragment of TTR, TTR81–127, is highly amyloidogenic in vitro, even at neutral pH. This fragment formed amyloid deposits and induced apoptosis and inflammatory gene expression also in cultured cells. Using the highly amyloidogenic TTR81–127 fragment, we developed a cell-based high-throughput screening method to discover compounds that disrupt TTR amyloid fibrils. Screening a library of 1280 off-patent drugs, we identified two candidate repositioning drugs, pyrvinium pamoate and apomorphine hydrochloride. Both drugs disrupted patient-derived TTR amyloid fibrils ex vivo, and pyrvinium pamoate also stabilized the tetrameric structure of TTR ex vivo in patient plasma. We conclude that our TTR81–127–based screening method is very useful for discovering therapeutic drugs that directly disrupt amyloid fibrils. We propose that repositioning pyrvinium pamoate and apomorphine hydrochloride as TTR amyloid-disrupting agents may enable evaluation of their clinical utility for managing ATTR amyloidosis.
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    Patterns of drug screen results and court-ordered substance use treatment referrals and completion among justice-involved youth
    (Elsevier, 2020-11) Dir, Allyson L.; Clifton, Richelle L.; Magee, Lauren A.; Johnson-Kwochka, Annalee V.; Wiehe, Sarah E.; Aalsma, Matthew C.; Psychiatry, School of Medicine
    Background Substance use is prevalent among justice-involved youth and given the risk of recidivism and other poor outcomes associated with substance use, justice systems have implemented efforts to improve substance use screening and connection to treatment. Although many justice systems use drug screening to monitor substance use, research on patterns of substance use based on drug screen records is lacking. The current study examined court records of drug screens among youth to explore patterns of substance use as well as rates of court-ordered referral to substance use treatment and treatment completion. We also examined differences in these patterns of use and treatment referral and completion by race, ethnicity, and gender. Method We examined court records for N = 3440 youth with records of positive oral drug screen (ODS) between 2011 and 2016 to assess patterns of ODS results (e.g., number and of positive screens), court-ordered referrals to substance use treatment, and rates of treatment completion. Results Of 3440 youth with a positive ODS, 96% tested positive for cannabis and 9.8% for opioids at least once; 48.5% were court-ordered to substance use treatment. Of those referred, 67% had history of completing at least one treatment episode; black youth ( OR = 0.54, p < .01) were less likely to have history of completing substance use treatment. Conclusion Our results underscore the need to utilize objective measures as well as validated self-reports of substance use history in both research and justice system decision-making to aid in identifying youth in need of services. Additional research should identify barriers to substance use treatment completion among this population.
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    Pressure sensitive adhesives and paper spray-mass spectrometry for the collection and analysis of fentanyl-related compounds from shipping materials
    (Wiley, 2023-09) Prunty, Sarah; Carmany, Daniel; Dhummakupt, Elizabeth S.; Manicke, Nicholas E.; Chemistry and Chemical Biology, School of Science
    The rise of fentanyl and fentanyl analogs in the drug supply pose serious threats to public health. Much of these compounds enter the United States through shipping routes. Here we provide a method for fentanyl screening and analysis that utilizes pressure-sensitive adhesive (PSA) lined paper to recover drug residues from parcel-related surfaces. The paper used is commercially available repositionable notes (also called post-it or sticky notes). From this paper, mass spectra were obtained by paper spray-mass spectrometry (PS-MS), where PSA paper served as both a sampling and analysis substrate. Seven fentanyl-related compounds were analyzed: fentanyl, 4-anilino-N-phenethylpiperidine (4-ANPP), N,1-diphenethyl-N-phenylpiperidin-4-amine (phenethyl-4-ANPP), valerylfentanyl, 4-fluoroisobutyrylfentanyl (4-FIBF), carfentanil, and p-fluorofentanyl. These compounds were recovered by PSA paper and identified by PS-MS from packaging tape and plastic at 50 ng and from cardboard and shipping labels at 100 ng. The impact of cutting agents on PS-MS analysis of fentanyl analogs was explored. No trends of analyte suppression were found at high concentrations of the cutting agents caffeine, diphenhydramine, and lidocaine when recovered from surfaces. A cartridge that required no precise cutting of PSA paper prior to sampling or analysis was evaluated for use in PS-MS for fentanyl screening. Recovery and detection of fentanyl from plastic sheeting was demonstrated with this cut-free cartridge. The cut-free cartridge showed somewhat less consistency and lower analyte signal than the standard cartridge, but performance was suitable for potential screening applications. In combining PSA surface sampling with PS-MS for drug screening, both sampling and detection of fentanyl-related compounds is simple, rapid, and low-cost.
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    Separation of Opiate Isomers using Electrospray Ionization and Paper Spray Coupled to High-Field Asymmetric Waveform Ion Mobility Spectrometry
    (Springer, 2015-05) Manicke, Nicholas E.; Belford, Michael; Department of Chemistry & Chemical Biology, IU School of Science
    One limitation in the growing field of ambient or direct analysis methods is reduced selectivity caused by the elimination of chromatographic separations prior to mass spectrometric analysis. We explored the use of high-field asymmetric waveform ion mobility spectrometry (FAIMS), an ambient pressure ion mobility technique, to separate the closely related opiate isomers of morphine, hydromorphone, and norcodeine. These isomers cannot be distinguished by tandem mass spectrometry. Separation prior to MS analysis is, therefore, required to distinguish these compounds, which are important in clinical chemistry and toxicology. FAIMS was coupled to a triple quadrupole mass spectrometer, and ionization was performed using either a pneumatically assisted heated electrospray ionization source (H-ESI) or paper spray, a direct analysis method that has been applied to the direct analysis of dried blood spots and other complex samples. We found that FAIMS was capable of separating the three opiate structural isomers using both H-ESI and paper spray as the ionization source.
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    Toxicological Drug Screening using Paper Spray High-Resolution Tandem Mass Spectrometry (HR-MS/MS)
    (Oxford, 2018-06) McKenna, Josiah; Jett, Rachel; Shanks, Kevin; Manicke, Nicholas E.; Chemistry and Chemical Biology, School of Science
    Immunoassays and high-performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) are both widely used methods for drug screening in toxicology. We investigated an alternative approach for rapid drug screening: paper spray MS (PS-MS). In paper spray, the biofluid sample is spotted onto a paper substrate. Upon application of a spray solvent and an electric potential, extraction and ionization occur directly from the paper without any need for additional sample preparation. We developed two paper spray high-resolution MS/MS targeted drug screening assays using a quadrupole-orbitrap mass spectrometer, one the positive ion mode and one in the negative ion mode. In the positive ion mode, over 130 drugs and drug metabolites were semi-quantitatively screened at sub-toxic concentrations in a single 2.5 min analysis. Limits of detection and calibration performances for each target compound are reported. The PS-MS/MS assay was tested on authentic postmortem specimens, and its screening ability and semi-quantitative performance were evaluated against independent LC–MS-MS screening and confirmation assays with good agreement. The paper spray MS/MS showed good qualitative agreement with LC–MS-MS; the true positive rate of paper spray MS/MS was 92%, and the true negative rate was over 98%. The quantitative results between the two methods were also acceptable for a screening application; Passing-Bablok regression yielded a slope of 1.17 and a Pearson’s correlation coefficient of 0.996. A separate PS-MS/MS negative ion screening method was also developed for a small panel of barbiturates and structural analogs, demonstrating its potential for acidic drug detection and screening.