Clinical Concentration Range Research Articles

Demyelination detection in CSF based on electrochemical monitoring of myelin basic protein in comparison between Apta vs. Immuno sensing strategies

Multiple sclerosis (MS) is a recurrent inflammatory, demyelinating disease of the white matter in central nervous system (CNS). The number of MS patients is increasing, but the diagnostic process is still quite difficult, costly and requires combination of several methods. Myelin basic protein (MBP) makes up to 30 % of the myelin in CNS. It is known that MBP is released into the cerebrospinal fluid (CSF) as MS bioindicator. Herein, myelin specific DNA aptamer earlier developed for possible therapeutic purposes and anti-MBP antibody were applied as bioreceptors for MBP recognition on the same nanomodified sensor surfaces and their performances were compared. Biosensors were developed by using graphene oxide (GO) nanoparticles integrated onto pencil graphite electrodes (PGE) and bioreceptor molecules immobilized to create a bioactive layer for MBP binding. The measurements were run with electrochemical impedance spectroscopy (EIS). Selectivity of the biosensors was evaluated using human serum albumin (HSA). After optimization of binding parameters, biosensors were validated in artificial CSF. It was shown that LJM-5708 based aptasensor had LOD 0.65 ng/mL that was comparable to immunosensor LOD (0.36 ng/mL) in artificial CSF and showed its applicability in the clinical concentration range between 1 and 128 ng/mL.

A-348 Portable aptamer-based home monitoring system for blood phenylalanine measurement

Abstract Background Aptatek BioSciences, Inc. has developed a home monitoring tool for blood phenylalanine (phe) levels. The Aptatek system integrates a proprietary aptamer assay approach with a user-friendly handheld, smartphone-enabled test platform. PKU (phenylketonuria), is an inborn error of phenylalanine (Phe) metabolism If undiagnosed or untreated, phe accumulates in all body tissues, including the blood and brain, where it is especially neurotoxic and when untreated it can cause significant cognitive impairment and neurological defects. PKU management targets maintaining blood Phe concentration within the therapeutic range (120-360uM) to avoid severe neurological sequelae, mainly through nutritional therapy. Methods PheCheck™ includes a portable hand-held reader, software accessible as a mobile phone application, and a disposable single-use lateral flow cassette, blood collection, cap, and buffer chamber. PheCheck™ detects phe by binding of the phe in a small blood sample using small molecule-specific DNA aptamers. A patient (or caregiver) collects 5μl of capillary blood using a lancet to lance the fingertip and use the collection cap to collect the specimen. Once collected, the specimen is placed into the buffer chamber where the blood is diluted and processed in preparation for a lateral flow assay. The buffer chamber is placed on top of a lateral flow cassette to release the solution into the cassette. The lateral flow cassette is inserted into a small electronic reader after specimen incubation. The reader will detect a fluorescent signal from the strip and report the quantitative Phe concentration present in the specimen to the integrated mobile phone application. The reader has a simple design with a single operating push button, USB data connection port, and replaceable battery compartment. The reader reads the output of the lateral flow cassette and reports the result to a Smartphone app. The app is designed to either display the quantitative results or store the results without displaying the quantitative value. Results Twelve patients were trained on the PheCheck system and observed using the system for self-testing. Patients also prepared dried blood spot cards for comparative testing using LC-mass spectrometry. Patients were asked to return to the clinic within two weeks to retest. The blood phenylalanine levels measured by the PheCheck system were within targeted standard deviation of the dried blood spots demonstrating the feasibility of portable home-based testing for routine monitoring of blood phenylalanine levels in phenylketonuria patients. Such a portable system would enable real time monitoring and adjustment of diet or therapy for chronic diseases such as PKU. Conclusions Aptatek has developed a DNA aptamer based portable monitor for measuring blood phenylalanine levels. The system quantitatively measures phenylalanine from a 5-microliter finger stick blood sampler. Aptamer-based detection of phenylalanine provides reliable quantitation across a wide clinical concentration range and encouraging performance relative to gold standard testing of venous blood draw or dried blood spots. Analysis of samples from patients on Palynziq indicate that the PheCheck system could provide more reliable testing than venous blood draws. The system shows strong promise for routine home monitoring of blood phe levels by patients.

Olaris Global Panel (OGP): A Highly Accurate and Reproducible Triple Quadrupole Mass Spectrometry-Based Metabolomics Method for Clinical Biomarker Discovery.

Mass spectrometry (MS)-based clinical metabolomics is very promising for the discovery of new biomarkers and diagnostics. However, poor data accuracy and reproducibility limit its true potential, especially when performing data analysis across multiple sample sets. While high-resolution mass spectrometry has gained considerable popularity for discovery metabolomics, triple quadrupole (QqQ) instruments offer several benefits for the measurement of known metabolites in clinical samples. These benefits include high sensitivity and a wide dynamic range. Here, we present the Olaris Global Panel (OGP), a HILIC LC-QqQ MS method for the comprehensive analysis of ~250 metabolites from all major metabolic pathways in clinical samples. For the development of this method, multiple HILIC columns and mobile phase conditions were compared, the robustness of the leading LC method assessed, and MS acquisition settings optimized for optimal data quality. Next, the effect of U-13C metabolite yeast extract spike-ins was assessed based on data accuracy and precision. The use of these U-13C-metabolites as internal standards improved the goodness of fit to a linear calibration curve from r2 < 0.75 for raw data to >0.90 for most metabolites across the entire clinical concentration range of urine samples. Median within-batch CVs for all metabolite ratios to internal standards were consistently lower than 7% and less than 10% across batches that were acquired over a six-month period. Finally, the robustness of the OGP method, and its ability to identify biomarkers, was confirmed using a large sample set.

Chemical stability of ondansetron hydrochloride with other drugs in admixtures via parenteral; a review

ObjectiveThis review was prepared to offer the most complete information about the use of ondansetron in parenteral admixtures with other drugs. MethodThe search was done from September 2016 to April 2017 by using electronic databases Stabilis® and Micromedex® solutions, Medli- ne/PubMed and Scholar Google searching publications about ondansetron stability in parenteral infusion when is administered by itself or with other medication. Results49 studies are included with a total of 53 drugs. 15 drugs were found compatible administered with ondansetron in a clinical routine concentration range in intravenous administration. Also, four ternary blends were found compatible and another one was incompatible. Otherwise, 38 drugs were found incompatible. ConclusionsCompatibility of ondansetron offers a broad number of options to be used to avoid nausea and vomiting symptoms in patients with other concomitant medication.

Neurocognitive effect of biased µ-opioid receptor agonist oliceridine, a utility function analysis and comparison with morphine.

Oliceridine (Olinvyk®) is a μ-opioid receptor agonist that in contrast to conventional opioids preferentially engages the G-protein-coupled signaling pathway. We determined the utility function of oliceridine versus morphine based on neurocognitive tests and cold pressor test. The study had a randomized, double-blind, placebo-controlled, partial block 3-way crossover design. Experiments were performed in 20 male and female volunteers. Subjects received intravenous oliceridine (1 or 3 mg; cohorts of 10 subjects/dose), morphine (5 or 10 mg; cohorts of 10 subjects/dose) or placebo on three separate occasions. Prior to and following dosing, neurocognitive tests, cold pressor test and plasma drug concentrations were obtained at regular intervals. Population pharmacokinetic-pharmacodynamic analyses served as basis for construction of a utility function, which is an objective function of probability of benefit minus probability of harm. Antinociception served as measure of benefit, slowing of saccadic peak velocity and increased body sway as measures of neurocognitive harm. The oliceridine and morphine C50's, i.e. the effect-site concentrations causing 50% effect, were: antinociception 13±2 and 23±7 ng/mL, saccadic peak velocity 90±14 and 54±15 ng/mL, and body sway 10±2 and 5.6±0.8 ng/mL, respectively. The ratio oliceridine/morphine of the therapeutic indices, C50(benefit)/C50(harm), were 0.34 (95% CI 0.17-0.7; p<0.01) for saccadic peak velocity and 0.33 (0.16-0.50; p<0.01) for body sway. The oliceridine utility was positive across the effect-site concentration 5-77 ng/mL, indicative of a greater probability of benefit than harm. The morphine utility was not significantly different from zero from 0-100 ng/mL. Over the concentration range 15-50 ng/mL the oliceridine utility was superior to that of morphine (p < 0.01). Similar observations were made for body sway. These data indicate that over the clinical concentration range, oliceridine is an analgesic with a favorable safety profile over morphine when considering analgesia and neurocognitive function.

Fluorescent immunochromatographic test strip for therapeutic drug monitoring of methotrexate with high sensitivity and wide dynamic range.

As a front-line chemotherapeutic drug for maintenance and consolidation therapy, methotrexate (MTX) has widely been applied to treat various tumors and some inflammatory diseases. However, because of its severe toxicity ascribed to low selectivity, it is necessary to monitor therapeutic drugs in high-dose MTX therapeutic regimens to ensure treatment safety. In this work, we developed a fluorescent immunochromatographic test strip (FITS) for monitoring MTX by employing time-resolved fluorescent microspheres as signal probes. With a competitive immunoassay mode, the FITS for MTX shows a super-wide dynamic range of 10pM-10μM, covering the entire clinical therapeutic concentration range of MTX. Therapeutic drug monitoring of MTX can be achieved within 7min with high specificity, facilitating the timely rescue of drug poisoning led by high-dose MTX treatment. The methodwasemployedfor monitoring MTX in the spiked human serum, urine, and milk, showing acceptable recoveriesranging from 94.0 to 110.0%. The established FITS has been applied toMTX detection in serum obtained from high-dose MTX treatment. The results from FITS and enzyme multiplied immunoassay technique showed no significant difference, suggesting its reliability for usage in real biological samples. The device shows promise in point-of-caretherapeutic drug monitoring for resource-limited countries and institutes, which significantly facilitates overcoming the lag time between sampling and results.

Oral Oxycodone-Induced Respiratory Depression During Normocapnia and Hypercapnia: A Pharmacokinetic-Pharmacodynamic Modeling Study.

The widely prescribed opioid oxycodone may cause lethal respiratory depression. We compared the effects of oxycodone on breathing and antinociception in healthy young volunteers. After pharmacokinetic/pharmacodynamic (PK/PD) modeling, we constructed utility functions to combine the wanted and unwanted end points into a single function. We hypothesized that the function would be predominantly negative over the tested oxycodone concentration range. Twenty-four male and female volunteers received 20 (n=12) or 40 (n=12) mg oral oxycodone immediate-release tablets. Hypercapnic ventilatory responses (visit 1) or responses to 3 nociceptive assays (pain pressure, electrical, and thermal tests; visit 2) were measured at regular intervals for 7 hours. the PK/PD analyses, that included carbon dioxide kinetics, stood at the basis of the utility function: probability of antinociception minus probability of respiratory depression. Oxycodone had rapid onset/offset times (30-40 minutes) with potency values (effect-site concentration causing 50% of effect) ranging from 0.05 to 0.13 ng/mL for respiratory variables obtained at hypercapnia and antinociceptive responses. Ventilation at an extrapolated end-tidal carbon dioxide partial pressure of 55 mmHg, was used for creation of 3 utility functions, one for each of the nociceptive tests. Contrary to expectation, the utility functions were close to zero or positive over the clinical oxycodone concentration range. The similar or better likelihood for antinociception relative to respiratory depression may be related to oxycodone's receptor activation profile or to is high likeability that possibly alters the modulation of nociceptive input. Oxycodone differs from other μ-opioids, such as fentanyl, that have a consistent negative utility.

Detection of the UV-vis silent biomarker trimethylamine-N-oxide via outer-sphere interactions in a lanthanide metal-organic framework

Trimethylamine-N-oxide (TMAO) is a biomarker of the cardiovascular disease that is one of the leading causes of worldwide death. Facile detection of TMAO can significantly improve the survival rate of this disease by allowing early prevention. However, the UV-vis silent nature of TMAO makes it intricated to be detected by conventional sensing materials or analytical instruments. Here we show a bilanthanide metal-organic framework functionalized by borono group for the recognition of TMAO. Superior sensitivity, selectivity and anti-interference ability were achieved by the inverse emission intensity changes of the two lanthanide centers. The limit of detection is 15.6 μM, covering the clinical urinary concentration range of TMAO. A smartphone application was developed based on the change in R-G-B chromaticity. The sensing mechanism via a well-matched outer-sphere interaction governing the sensing function was studied in detail, providing fundamentals in molecular level for the design of advanced sensing materials for UV-Vis silent molecules.

Silica embedded carbon nanosheets derived from biomass acorn cupule for non-enzymatic, label-free, and wide range detection of α 1-acid glycoprotein in biofluids

In this work, we demonstrate the first report on a low-cost, non-enzymatic, label-free electrochemical sensing of alpha-1-acid-glycoprotein (α1GP) biomarker in biofluids using silica embedded carbon nanosheets (SEC) derived from acorn cupules biomass. The SEC/GCE sensor exhibits low detection limit (LOD) of 30 ng/mL (LOD = 3 s/m) which is far below the lowest physiological concentration of α1GP and a wide linear detection range from 100 ng/mL to 10 mg/mL which covers entire clinical concentration range reported for various diseases like cancer, sepsis, etc. The sensing mechanism of the sensor relies on the direct electrooxidation of sialic acid from the α1GP structure which hinder the interfacial electron-transfer process of [Fe (CN)6]3-/4- redox probe at the electrode-electrolyte interface. The sensor exhibits excellent selectivity and stability towards detection of α1GP which is ascribed to the presence of embedded silica nanoparticles on the surface of the carbon nanosheets. The successful determination of α1GP in the simulated blood serum sample with good recovery percentage (i.e., from ∼94.03% to ∼103.50%) proves the feasibility of the sensor towards clinical applications. The overall efficacy of the sensor proves it as a promising low-cost, sustainable, and non-enzymatic platform for a wide variety of bioanalytical applications.

In-fiber optofluidic online SERS detection of trace uremia toxin.

In this Letter, we propose a microstructured in-fiber optofluidic surface-enhanced Raman spectroscopy (SERS) sensor for the initial inspection of uremia through the detection of unlabeled urea and creatinine. As a natural microfluidic device, microstructured hollow fiber has a special structure inside. Through chemical bonds, the SERS substrate can be modified and grown on the surface of the suspended core. Here, the silver nanoparticles (Ag NPs) are embedded on the poly diallyl dimethyl ammonium chloride-modified graphene oxide sheet to achieve the self-assembled SERS substrate. The reduced distance between Ag NPs can increase the strong hot spots that generate enhanced Raman signals. Therefore, it can effectively detect the Raman signal of unlabeled trace uremic toxin analytes (urea, creatinine) inside the optical fiber. The results show that under simulated biophysical conditions, the limit detection (LOD) for urea is 10-4M and the linearity is good, especially at the clinical conventional concentration range (2.5-6.5×10-3M). In addition, the online Raman detection of creatinine aqueous solution LOD is 10-6M, which also has good linearity. Significantly, this Letter provides a microstructured optofluidic in-fiber Raman sensor for the preliminary detection of uremia, which will have good development prospects in the field of clinical biomedicine.

Detection of antibodies against SARS-CoV-2 spike protein by gold nanospikes in an opto-microfluidic chip

The ongoing global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to active research in its associated diagnostics and medical treatments. While quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most reliable method to detect viral genes of SARS-CoV-2, serological tests for specific antiviral antibodies are also important as they identify false negative qRT-PCR responses, track how effectively the patient’s immune system is fighting the infection, and are potentially helpful for plasma transfusion therapies. In this work, based on the principle of localized surface plasmon resonance (LSPR), we develop an opto-microfluidic sensing platform with gold nanospikes, fabricated by electrodeposition, to detect the presence and amount of antibodies specific to the SARS-CoV-2 spike protein in 1μL of human plasma diluted in 1mL of buffer solution, within ∼30min. The target antibody concentration can be correlated with the LSPR wavelength peak shift of gold nanospikes caused by the local refractive index change due to the antigen–antibody binding. This label-free microfluidic platform achieves a limit of detection of ∼0.08ng/mL (∼0.5pM), falling under the clinical relevant concentration range. We demonstrate that our opto-microfluidic platform offers a promising point-of-care testing tool to complement standard serological assays and make SARS-CoV-2 quantitative diagnostics easier, cheaper, and faster.

Placental transfer of Letermovir & Maribavir in the ex vivo human cotyledon perfusion model. New perspectives for in utero treatment of congenital cytomegalovirus infection.

Congenital cytomegalovirus infection can lead to severe sequelae. When fetal infection is confirmed, we hypothesize that fetal treatment could improve the outcome. Maternal oral administration of an effective drug crossing the placenta could allow fetal treatment. Letermovir (LMV) and Maribavir (MBV) are new CMV antivirals, and potential candidates for fetal treatment. The objective was to investigate the placental transfer of LMV and MBV in the ex vivo method of the human perfused cotyledon. Term placentas were perfused, in an open-circuit model, with LMV or MBV at concentrations in the range of clinical peak plasma concentrations. Concentrations were measured using ultraperformance liquid chromatography coupled with tandem mass spectrometry. Mean fetal transfer rate (FTR) (fetal (FC) /maternal concentration), clearance index (CLI), accumulation index (AI) (retention of each drug in the cotyledon tissue) were measured. Mean FC were compared with half maximal effective concentrations of the drugs (EC50(LMV) and EC50(MBV)). For LMV, the mean FC was (± standard deviation) 1.1 ± 0.2 mg/L, 1,000-fold above the EC50(LMV). Mean FTR, CLI and AI were 9 ± 1%, 35 ± 6% and 4 ± 2% respectively. For MBV, the mean FC was 1.4 ± 0.2 mg/L, 28-fold above the EC50(MBV). Mean FTR, CLI and AI were 10 ± 1%, 50 ± 7% and 2 ± 1% respectively. Drugs' concentrations in the fetal side should be in the range for in utero treatment of fetuses infected with CMV as the mean FC was superior to the EC50 for both molecules.

Testing of the Survivin Suppressant YM155 in a Large Panel of Drug-Resistant Neuroblastoma Cell Lines.

The survivin suppressant YM155 is a drug candidate for neuroblastoma. Here, we tested YM155 in 101 neuroblastoma cell lines (19 parental cell lines, 82 drug-adapted sublines). Seventy seven (77) cell lines displayed YM155 IC50s in the range of clinical YM155 concentrations. ABCB1 was an important determinant of YM155 resistance. The activity of the ABCB1 inhibitor zosuquidar ranged from being similar to that of the structurally different ABCB1 inhibitor verapamil to being 65-fold higher. ABCB1 sequence variations may be responsible for this, suggesting that the design of variant-specific ABCB1 inhibitors may be possible. Further, we showed that ABCC1 confers YM155 resistance. Previously, p53 depletion had resulted in decreased YM155 sensitivity. However, TP53-mutant cells were not generally less sensitive to YM155 than TP53 wild-type cells in this study. Finally, YM155 cross-resistance profiles differed between cells adapted to drugs as similar as cisplatin and carboplatin. In conclusion, the large cell line panel was necessary to reveal an unanticipated complexity of the YM155 response in neuroblastoma cell lines with acquired drug resistance. Novel findings include that ABCC1 mediates YM155 resistance and that YM155 cross-resistance profiles differ between cell lines adapted to drugs as similar as cisplatin and carboplatin.

The immunomodulatory-drug, lenalidomide, sustains and enhances interferon-α production by human plasmacytoid dendritic cells.

Background: Lenalidomide (LEN), an immunomodulatory drug (IMiD), is currently used for treatment of multiple myeloma (MM). LEN potentiates T cell and natural killer cell functions. However, the cellular and molecular mechanisms underlying the immunomodulatory effects of LEN remain unclear. We focused on the effects of LEN on human plasmacytoid dendritic cells (pDCs), which are the major source of interferon (IFN)-α in the blood and play a central role in innate immune responses.Results: We found that bortezomib, a proteasome inhibitor used to treat MM, killed pDCs but that 0.1–3 μM LEN (covering clinical plasma concentration range) did not affect pDC survival or CD86 expression. Bortezomib inhibited pDC-derived IFN-α production in a dose-dependent fashion, but 0.1–3 µM LEN sustained pDC-derived IFN-α production when stimulated with an optimal concentration of CpG-ODN 2216 (3 μM). In pDCs stimulated with a low concentration of CpG-ODN (0.1 μM), LEN enhanced IFN-α production. These results indicated that LEN, when used at a clinically relevant concentration, can potentially enhance IFN-α production by pDCs.Conclusion: Collectively, our findings unveiled a novel target of LEN and extend the repertoire of the drug’s known immunomodulatory effects. These effects may explain the low incidence of herpes zoster viral infection observed during LEN treatment compared with bortezomib treatment. LEN may function as an IMiD affecting a wide array of immune cells, including pDCs, leading to amplification of a positive immune axis able to eliminate MM cells.

Ketamine Within Clinically Effective Range Inhibits Glutamate Transmission From Astrocytes to Neurons and Disrupts Synchronization of Astrocytic SICs.

BackgroundAstrocytes are now considered as crucial modulators of neuronal synaptic transmission. General anesthetics have been found to inhibit astrocytic activities, but it is not clear whether general anesthetics within the clinical concentration range affects the astrocyte-mediated synaptic regulation.MethodsThe effects of propofol, dexmedetomidine, and ketamine within clinically effective ranges on the slow inward currents (SICs) were tested by using the whole-cell recording in acute prefrontal cortex (PFC) slice preparations of rats. Astrocytes culture and HPLC were used to measure the effects of different anesthetics on the glutamate release of astrocytes.ResultsPropofol and dexmedetomidine showed no significant effect on the amplitude or frequency of SICs. Ketamine was found to inhibit the frequency of SICs in a concentration-dependent manner. The SICs synchronization rate of paired neurons was inhibited by 30 μM ketamine (from 42.5 ± 1.4% to 9.6 ± 0.8%) and was abolished by 300 μM ketamine. The astrocytic glutamate release induced by DHPG, an agonist of astrocytic type I metabotropic glutamate receptors, was not affected by ketamine, and ifenprodil, a selective antagonist of GluN1/GluN2B receptor, blocked all SICs and enhanced the inhibitory effect of 30 μM ketamine on the frequency of SICs. Ketamine at low concentration (3 μM) could inhibit the frequency of SICs, not the miniature excitatory postsynaptic currents (mEPSCs), and the inhibition rate of SICs was significantly higher than mEPSCs with 30 μM ketamine (44.5 ± 3% inhibition vs. 28.3 ± 6% inhibition).ConclusionOur data indicated that ketamine, not propofol and dexmedetomidine, within clinical concentration range inhibits glutamatergic transmission from astrocytes to neurons, which is likely mediated by the extrasynaptic GluN1/GluN2B receptor activation.

Quantum Dot-Based FRET Immunoassay for HER2 Using Ultrasmall Affinity Proteins.

Engineered scaffold affinity proteins are used in many biological applications with the aim of replacing natural antibodies. Although their very small sizes are beneficial for multivalent nanoparticle conjugation and efficient Förster resonance energy transfer (FRET), the application of engineered affinity proteins in such nanobiosensing formats has been largely neglected. Here, it is shown that very small (≈6.5 kDa) histidine-tagged albumin-binding domain-derived affinity proteins (ADAPTs) can efficiently self-assemble to zwitterionic ligand-coated quantum dots (QDs). These ADAPT-QD conjugates are significantly smaller than QD-conjugates based on IgG, Fab', or single-domain antibodies. Immediate applicability by the quantification of the human epidermal growth factor receptor 2 (HER2) in serum-containing samples using time-gated Tb-to-QD FRET detection on the clinical benchtop immunoassay analyzer KRYPTOR is demonstrated here. Limits of detection down to 40 × 10-12 m (≈8 ng mL-1 ) are in a relevant clinical concentration range and outperform previously tested assays with antibodies, antibody fragments, and nanobodies.

Simultaneous Determination of Five Cytochrome P450 Probe Substrates and Their Metabolites and Organic Anion Transporting Polypeptide Probe Substrate in Human Plasma Using Liquid Chromatography-Tandem Mass Spectrometry

A rapid and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of organic anion transporting polypeptide 1B1 (OATP1B1) and cytochrome P450 (P450) probe substrates and their phase I metabolites in human plasma was developed. The OATP1B1 (pitavastatin) and five P450 probe substrates, caffeine (CYP1A2), losartan (CYP2C9), omeprazole (CYP2C19), dextromethorphan (CYP2D6), and midazolam (CYP3A) and their metabolites were extracted from human plasma (50 µL) using methanol. Analytes were separated on a C18 column followed by selected reaction monitoring detection using MS/MS. All analytes were separated simultaneously within a 9 min run time. The developed method was fully validated over the expected clinical concentration range for all analytes tested. The intra- and inter-day precisions for all analytes were lower than 11.3% and 8.82%, respectively, and accuracy was 88.5–117.3% and 96.1–109.2%, respectively. The lower limit of quantitation was 0.05 ng/mL for dextromethorphan, dextrorphan, midazolam, and 1′-hydroxymidazolam; 0.5 ng/mL for losartan, EXP-3174, omeprazole, 5′-hydroxyomeprazole, and pitavastatin; and 5 ng/mL for caffeine and paraxanthine. The method was successfully used in a pharmacokinetic study in healthy subjects after oral doses of five P450 and OATP1B1 probes. This analytical method provides a simple, sensitive, and accurate tool for the determination of OATP1B1 and five major P450 activities in vivo drug interaction studies.

Quantitative analysis of prompt gamma ray imaging during proton boron fusion therapy according to boron concentration

The purpose of this study is to evaluate the prompt gamma ray imaging technique according to the clinical boron concentration range during proton boron fusion therapy (PBFT). To acquire a prompt gamma ray image from 32 projections, we simulated four head single photon emission computed tomography and a proton beam nozzle using a Monte Carlo simulation. We used modified ordered subset expectation maximization reconstruction algorithm with a graphic processing unit for fast image acquisition. Boron concentration was set as 20 to 100 μg at intervals of 20 μg. For quantitative analysis of the prompt gamma ray image, we acquired an image profile drawn through two boron uptake regions (BURs) and calculated the contrast value, signal-to-noise ratio (SNR), and difference between the physical target volume and volume of the prompt gamma ray image. The relative counts of prompt gamma rays were noticeably increased with increasing boron concentration. Although the intensities on the image profiles showed a similar tendency according to the boron concentration, the SNR and contrast value were improved with increasing boron concentration. This study suggests that a tumor monitoring technique using prompt gamma ray detection can be clinically applicable even if the boron concentration is relatively low.

Detection of Biomarkers of Periodontal Disease in Human Saliva Using Stabilized, Vertical Flow Immunoassays.

We report methods for stabilizing cellulose-based immunoassays and using this platform to analyze human saliva. Stabilization treatments of immunoassays for matrix metalloproteinases (MMP)-8 and -9, biomarkers of periodontal disease, were conducted and compared, revealing that anti-MMP-8 and -9 capture antibodies could be stabilized with the addition of a 5% trehalose solution to the test zones, followed by drying in a vacuum oven. After stabilization, the paper devices retained equivalent binding activity to that of freshly prepared tests for 14 days-a time frame that enables US-based clinical testing of this diagnostic assay. A saliva pretreatment method was developed to remove viscous elements without reducing the concentration or binding activity of dissolved proteins. Immunoassays were stored in ziplock bags containing desiccant, and used to detect nanomolar concentrations of MMP-9 in human saliva across the relevant clinical concentration range. These methods and findings facilitate rapid, affordable validation studies of this and other biomarkers that are found in saliva using vertical flow immunoassays.

Quantitative comparison of PET performance-Siemens Biograph mCT and mMR.

BackgroundIntegrated clinical whole-body PET/MR systems were introduced in 2010. In order to bring this technology into clinical usage, it is of great importance to compare the performance with the well-established PET/CT. The aim of this study was to evaluate PET performance, with focus on image quality, on Siemens Biograph mMR (PET/MR) and Siemens Biograph mCT (PET/CT).MethodsA direct quantitative comparison of the performance characteristics between the mMR and mCT system was performed according to National Electrical Manufacturers Association (NEMA) NU 2-2007 protocol. Spatial resolution, sensitivity, count rate and image quality were evaluated. The evaluation was supplemented with additional standardized uptake value (SUV) measurements.ResultsThe spatial resolution was similar for the two systems. Average sensitivity was higher for the mMR (13.3 kcps/MBq) compared to the mCT system (10.0 kcps/MBq). Peak noise equivalent count rate (NECR) was slightly higher for the mMR (196 kcps @ 24.4 kBq/mL) compared to the mCT (186 kcps @ 30.1 kBq/mL). Scatter fractions in the clinical activity concentration range yielded lower values for the mCT (34.9 %) compared to those for the mMR (37.0 %). Best image quality of the systems resulted in approximately the same mean hot sphere contrast and a difference of 19 percentage points (pp) in mean cold contrast, in favour of the mCT. In general, point spread function (PSF) increased hot contrast and time of flight (TOF) increased both hot and cold contrast. Highest hot contrast for the smallest sphere (10 mm) was achieved with the combination of TOF and PSF on the mCT. Lung residual error was higher for the mMR (22 %) than that for the mCT (17 %), with no effect of PSF. With TOF, lung residual error was reduced to 8 % (mCT). SUV was accurate for both systems, but PSF caused overestimations for the 13-, 17- and 22-mm spheres.ConclusionsBoth systems proved good performance characteristics, and the PET image quality of the mMR was close to that of the mCT. Differences between the systems were mainly due to the TOF possibility on the mCT, which resulted in an overall better image quality, especially for the most challenging settings with higher background activity and small uptake volumes.