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Quantification of coronary artery stenosis on X-ray angiography (XRA) images is of great importance during the intraoperative treatment of coronary artery disease. It serves to quantify the coronary artery stenosis by estimating the clinical morphological indices, which are essential in clinical decision making. However, stenosis quantification is still a challenging task due to the overlapping, diversity and small-size region of the stenosis in the XRA images. While efforts have been devoted to stenosis quantification through low-level features, these methods have difficulty in learning the real mapping from these features to the stenosis indices. These methods are still cumbersome and unreliable for the intraoperative procedures due to their two-phase quantification, which depends on the results of segmentation or reconstruction of the coronary artery. In this work, we are proposing a hierarchical attentive multi-view learning model (HEAL) to achieve a direct quantification of coronary artery stenosis, without the intermediate segmentation or reconstruction. We have designed a multi-view learning model to learn more complementary information of the stenosis from different views. For this purpose, an intra-view hierarchical attentive block is proposed to learn the discriminative information of stenosis. Additionally, a stenosis representation learning module is developed to extract the multi-scale features from the keyframe perspective for considering the clinical workflow. Finally, the morphological indices are directly estimated based on the multi-view feature embedding. Extensive experiment studies on clinical multi-manufacturer dataset consisting of 228 subjects show the superiority of our HEAL against nine comparing methods, including direct quantification methods and multi-view learning methods. The experimental results demonstrate the better clinical agreement between the ground truth and the prediction, which endows our proposed method with a great potential for the efficient intraoperative treatment of coronary artery disease. [ABSTRACT FROM AUTHOR]

The integration of segmentation and direct quantification on the left ventricle (LV) from the paired apical views(i.e., apical 4-chamber and 2-chamber together) echo sequence clinically achieves the comprehensive cardiac assessment: multiview segmentation for anatomical morphology, and multidimensional quantification for contractile function. Direct quantification of LV, i.e., to automatically quantify multiple LV indices directly from the image via task-aware feature representation and regression, avoids accumulative error from the inter-step target. This integration sequentially makes a stereoscopical reflection of cardiac activity jointly from the paired orthogonal cross views sequences, overcoming limited observation with a single plane. We propose a K-shaped Unified Network (K-Net), the first end-to-end framework to simultaneously segment LV from apical 4-chamber and 2-chamber views, and directly quantify LV from major- and minor-axis dimensions (1D), area (2D), and volume (3D), in sequence. It works via four components: 1) the K-Net architecture with the Attention Junction enables heterogeneous tasks learning of segmentation task of pixel-wise classification, and direct quantification task of image-wise regression, by interactively introducing the information from segmentation to jointly promote spatial attention map to guide quantification focusing on LV-related region, and transferring quantification feedback to make global constraint on segmentation; 2) the Bi-ResLSTMs distributed in K-Net layer-by-layer hierarchically extract spatial-temporal information in echo sequence, with bidirectional recurrent and short-cut connection to model spatial-temporal information among all frames; 3) the Information Valve tailing the Bi-ResLSTMs selectively exchanges information among multiple views, by stimulating complementary information and suppressing redundant information to make the efficient cross-flow for each view; 4) the Evolution Loss comprehensively guides sequential data learning, with static constraint for frame values, and dynamic constraint for inter-frame value changes. The experiments show that our K-Net gains high performance with a Dice coefficient up to 91.44% and a mean absolute error of the major-axis dimension down to 2.74mm, which reveal its clinical potential. [ABSTRACT FROM AUTHOR]

• Direct PCR amplification is comparable to conventional DNA profiling. • Identifiler® Plus at 29 cycles, half volume reaction with TE buffer was optimal. • 1 × 1 mm2 and 2 × 1 mm2 sample size is recommended. • Expected mixture ratios not conserved following direct amplification. • Direct quantitation shown to be a qualitative tool. [ABSTRACT FROM AUTHOR]

Ambient ionization in open environment brings a capability of a coupled mass spectrometry to detect target molecules in situ. However, it is limited to qualitative and semi-quantitative analysis. By coupling of an ambient ionization-based Direct Analysis in Real Time (DART) with high-resolution quadrupole time-of-flight mass spectrometry (QTOF/MS), we observe that, in one-chemical system, the target molecule displays a non-linear response in MS signal vs concentration, accompanying with large variation in MS signal, suggesting two obstacles for quantification to be overcome. Surprisingly, in a two-chemical system, we observe an apparent suppression effect. We prove that, due to this observed suppression effect, a fluctuant response in the MS signal of the stable isotope-labeled analogue can immediately reflect the change in the analyte concentration and ionization efficiency. For example, by taking advantage of this effect, even the analyte of different concentrations despairingly displayed similar signals would be accurately calibrated through the suppression of the internal stable isotope standard. This puts an important foundation on accurate and linear quantitation of analytes in complex matrix using DART-MS assay. Moreover, we for the first time demonstrate an application of in situ calibration of DART-MS for direct and accurate quantification of target molecule (creatinine) in highly complex samples (human urine) without any pre-separation. The quantification is also validated using HPLC-UV analysis (n = 38). At last, we show that stable isotope-labeled-creatinine (m / z 117.0850 amu) can be used for simultaneous in situ calibration of some other urinary metabolites with a mass/charge ratio varying from 120.069 amu to 333.125 amu. Image 1 • DART-QTOF/MS method is developed for direct and accurate quantification of chemicals in complex matrix such as urine. • Urine excretion rate index creatinine is used as a representative example of target molecule. • Stable isotope-labeled creatinine ([ D 3-creatinine]) is used for accurate in situ calibration. • Several other urinary metabolites can be simultaneously calibrated only using stable isotope-labeled creatinine. [ABSTRACT FROM AUTHOR]

Circulating microRNAs (miRNAs) are considered as reliable candidates for biomarker discovery. RNA-Sequencing has become the most suitable technique to accurately quantify the miRNAome. However, RNA-Sequencing relies on several technical passages before reaching the final-end. HTG EdgeSeq technology, thanks to the abrogation of RNA extraction step, allows productivity enhancement by reducing the number of hands-on steps, the time for sample preparation and, therefore, the costs. We found a strong correlation between qPCR and dPCR with HTG (Pearson’s coefficient of 0.93 and 0.94, respectively). In conclusion, we showed that HTG EdgeSeq, performed on human plasma specimens without RNA extraction, is reliable, allows the simultaneous screening of more than 2,000 miRNAs, and thus, it could be applied to biomarker discovery in large cohorts. [ABSTRACT FROM AUTHOR]

Quantitative PCR by directly sampling (Cells-qPCR) has been adapted to detect and quantify total yeasts, and B. bruxellensis, S. cerevisiae and Z. bailii species, in grape musts and wines. To increase assay sensitivity, the effects of a previous cell wall lysis, by both enzymatic and mechanical methods, were evaluated. Cell wall disruption by mechanical methods showed the best results to enhance assay sensitivity. Numerous standard curves were constructed by mechanically lysed cells in culture medium, and in white and red grape musts and wines. Good regression values (>0.99) and efficiencies (>0.99) were obtained, and it was possible to detect one single cell per reaction in all the matrices. Moreover, the population evolution of yeasts during the winemaking process showed that the method provides an effective tool to detect and enumerate yeasts during industrial wine fermentation, and to rapidly control wine spoilage risks. [ABSTRACT FROM AUTHOR]

Falsified drugs are a threat to the health of patients. The analytical control of such products contributes to the fight against this global issue. Raman chemical imaging is a method that relies on consecutive measurements at the surface of a sample, combining spectroscopy, microscopy and chemometrics. This article explores the capabilities of this analytical technique proposing an innovative methodology with spectroscopic screening for the identification of chemical compounds and the direct quantification of the active substance (without prior calibration). Two chemometric methods were used: Multivariate Curve Analysis − Alternate Least Squares for the qualitative analysis and Direct Classical Least Squares for the quantitative analysis. The methodology was optimized with samples prepared in the laboratory and validation parameters were studied. The methodology was then applied to real (authentic and falsified) samples of Viagra ® and Plavix ® . Despite the presence of fluorescence emission in some samples, the methodology succeeded in the detection of active pharmaceutical ingredients, and in the discrimination of three salts of clopidogrel (in generic formulations of Plavix ® ). The quantitative deviation from the reference method ranged from −15% to +24% of the active substance content. This deviation may be considered to be acceptable since it is sufficient for assessing the risk to the health of patients and for quickly alerting the health authorities. [ABSTRACT FROM AUTHOR]

In this paper, we present a compact Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS) designed for real time analysis of volatile organic compounds (VOCs) in air or in water. The spectrometer is based on a structured permanent magnet made with NdFeB segments. Chemical ionization is implemented inside the ICR cell. The most widely used reaction is the proton transfer reaction using H3O+ precursor ions, but other ionic precursors can be used to extend the range of species that can be detected. Complex mixtures are studied by switching automatically from one precursor to another. The accuracy obtained on the mass to charge ratio (Δm/z 5 × 10−3), allows a precise identification of the VOCs present and the limit of detection is 200 ppb without accumulation. The time resolution is a few seconds, mainly limited by the time necessary to come back to background pressure after the gas pulses. The real time measurement will be illustrated by the monitoring of VOCs produced during the thermal degradation of a polymer and by an example where three different precursor ions are used alternatively to monitor a gas sample.

DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration material and it has higher tolerance to inhibitors. DNA quantification without an extraction step (i.e. direct quantification) was performed here using dPCR and two different human cytomegalovirus whole-virus materials. Two dPCR platforms were used for this direct quantification of the viral DNA, and these were compared with quantification of the extracted viral DNA in terms of yield and variability. Direct quantification of both whole-virus materials present in simple matrices like cell lysate or Tris-HCl buffer provided repeatable measurements of virus concentrations that were probably in closer agreement with the actual viral load than when estimated through quantification of the extracted DNA. Direct dPCR quantification of other viruses, reference materials and clinically relevant matrices is now needed to show the full versatility of this very promising and cost-efficient development in virus quantification. [ABSTRACT FROM AUTHOR]

Abstract: The current short tandem repeat (STR) typing workflow for forensic casework samples involves sample collection, sample screening, DNA extraction, DNA qPCR quantification and STR amplification. Although very effective and powerful, this workflow still has room for improvements. For example, the screening assays in practice do not provide DNA related information and also do not work with touch DNA samples, which make up of the majority of the property crime samples. It is known that not all DNA samples have equal probative values. Considering the DNA backlog situation crime laboratories face today, an effective screening tool would be highly desirable. It would allow forensic scientists to prioritize the DNA samples so that the limited resources would be first spent on samples that would have better chances of producing informative STR profiles. qPCR assay does provide DNA quantity and gender information and would be an ideal screening tool. However, prior to quantification, sample extraction and purification are required. By the time a DNA sample is ready for qPCR quantification, time and resources have already been spent on samples that should have been given low priority or excluded from further processing if DNA quantity and gender information were known. To overcome this problem, a direct quantification technology is developed to allow qPCR quantification of casework samples without the need for DNA extraction and purification. The key to a direct qPCR assay is the PE-Swab, a novel sample collection device. A small sample punch can be generated from a PE-Swab and placed in a qPCR reaction for quantification. After optimizing the punch size and the quantification software baseline setting, accurate DNA quantification can be obtained from a sample without the need to carry out DNA extraction and purification. Proof of concept studies were done with low lever touch samples as well as blood samples. The PE-Swab also allows direct STR amplification of casework samples without the need for DNA extraction. Besides its potential as a screening tool, the direct qPCR assay can also be used to normalize the DNA input for a direct STR amplification reaction. The feasibility of the direct qPCR/direct STR amplification workflow was demonstrated with touch DNA samples and blood stain samples. [Copyright &y& Elsevier]

Highlights: [•] Improvement of the direct quantification with analytical pyrolysis. [•] Optimization of the chromatographic parameters after pyrolysis of cationic starch. [•] Quantification of cationic starches in cellulose matrixes. [•] Validation of the cationic starch quantification in paper samples. [ABSTRACT FROM AUTHOR]

Abstract: This review primarily deals with the compensation of ‘matrix effect’ in secondary ion mass spectrometry (SIMS) for direct quantitative analysis of materials using MCs + -SIMS approach. Emphasis has been given on exploring the formation mechanisms of MCsn + (n = 1, 2,..) molecular ions (M denotes the element to be analyzed and Cs+ is the bombarding ion) emitted in the SIMS process. Following a brief introduction on SIMS, a study on MCsn + molecular ions emitted from various metal targets under Cs+ primary bombardment has been discussed. These studies essentially concentrate on the secondary ion energy distributions of the emitted species under different bombarding energies and impact angles. We have discussed a systematic study on the effect of reactive species like oxygen and cesium on the emission of MCsn + molecular ions. In all cases we have estimated the changes in local surface work function from the leading parts of the secondary ion energy distributions. The observations are explained in the light of surface work function changes at the sputtering sites. Formation mechanisms of MCsn + molecular ions are explained in the framework of sputter-ion emission models. Suppression of matrix effects in the compositional analysis of molecular beam epitaxy (MBE) - grown SiGe alloy structures has been discussed in detail. We have essentially applied the MCs + -SIMS method for the minimization of ‘matrix effect’, which is routinely encountered in conventional SIMS analysis hindering quantification of materials. Under the impact of energetic Cs+ ions, both SiCs+ and GeCs+ ions have been monitored for SiGe alloy samples with Ge concentration between 0 and 72 at.%. Following optimization of various experimental parameters, we have shown that the matrix effects are practically suppressed for Ge content up to 72 at.% in SiGe samples. A procedure for the accurate quantification of Ge concentration in Si1−xGex alloys using this method has been presented. This novel methodology has successfully been applied in the depth profiling of Si/Ge multilayer structures grown by MBE technique. [Copyright &y& Elsevier]

Most pesticides, herbicides and other plant treatment agents are applied to the crop surface. Direct mass spectrometric methods, such as desorption electrospray ionization (DESI), offer new ways to analyze plant samples directly and rapidly. A strategy for the development and optimization of a DESI method for the direct determination of chemicals on complex surfaces is described. Chlorpropham (CP) was applied to potato surfaces as an example for a crop protection agent and analyzed using a self-made DESI source. Aspects such as instrument selectivity, sensitivity and reproducibility were investigated. The MS4 fragmentation pattern of CP was analyzed to achieve the necessary detection selectivity, and is discussed in detail. Similar fragmentation was found in the ESI and DESI mass spectra, indicating that the mechanisms of ESI and DESI are closely related. A DESI method for semi-quantification of CP on potatoes was developed. Detection limits of 6.5 µg/kg were found using MS/MS. The reproducibility, in the range of 12% (signal variation), appears to be sufficient for semi-quantitative measurements. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: This article analyses the interferences in the quantification of traces of bisphenol S in paper by applying the direct analytical method “analytical pyrolysis gas chromatography mass spectrometry” (Py-GC/MS) in conjunction with on-line derivatisation with tetramethylammonium hydroxide (TMAH). As the analytes are simultaneously analysed with the matrix, the interferences derive from the matrix. The investigated interferences are found in the analysis of paper samples, which include bisphenol S derivative compounds. As the free bisphenol S is the hydrolysis product of the bisphenol S derivative compounds, the detected amount of bisphenol S in the sample may be overestimated. It is found that the formation of free bisphenol S from the bisphenol S derivative compounds is enhanced in the presence of tetramethylammonium hydroxide (TMAH) under pyrolytic conditions. In order to avoid the formation of bisphenol S trimethylsulphonium hydroxide (TMSH) is introduced. Different parameters are optimised in the development of the quantification method with TMSH. The quantification method based on TMSH thermochemolysis has been validated in terms of reproducibility and accuracy. [Copyright &y& Elsevier]

Multiplex real-time PCR assays were developed to quantify multiple species of Meloidogyne incognita, Pratylenchus penetrans, Globodera rostochiensis and Heterodera glycines in soil. The probes specific for P. penetrans and H. glycines are labelled with a fluorescence molecule, FAM, and those for M. incognita and G. rostochiensis with ROX. The primers and probes are species-specific to P. penetrans, but group-specific to the other species. DNA was extracted from suspensions containing each nematode and multiplex Cycleave® PCR assays were done for pairs of P. penetrans and M. incognita, P. penetrans and G. rostochiensis, or G. rostochiensis and H. glycines. The results revealed that the target nematode, except for H. glycines, was quantified in the presence of less than 100 times that of the other nematode (competitor), but underestimated in the presence of 1000 times the competitor. Such underestimation was solved by the use of SYBR Green I real time PCR assays targeting a single species. Multiplex PCR assay for P. penetrans and M. incognita was done using environmental DNA (eDNA) extracted from a soil naturally infested with the nematodes. Results quantified both species. Multiplex assay using eDNA may enable a sensitive and simultaneous detection of P. penetrans and M. incognita or P. penetrans and G. rostochiensis in soil although caution is needed in case the existing ratio is biased to one of the species. [ABSTRACT FROM AUTHOR]

Abstract: mRNA quantification has become a research hotspot. Quantitative real-time RT-PCR is a popular method but is known to lack precision. To rapidly monitor the kinetics of mRNA levels for the control of microbial fermentation processes, we developed an SYBR Green I-based universal method to directly quantify mRNA from fermentation samples. After total RNA was extracted, the mRNA was hybridized and protected by a longer DNA oligonucleotide. The probe length determined the strength of signal amplification. S1 nuclease and RNase A were used to remove excess probe, single-stranded RNA, and mis-matched RNA/DNA hybrids. Finally, the perfect-matched RNA/DNA hybrid was quantified by SYBR Green I dye. The conditions of liquid hybridization and enzyme digestion were systemically optimized. The kinetic tendency of phzC mRNA levels during phenazine-1-carboxylic acid fermentation was consistent with the results from MB hybridization in our previous report. The detection of mRNA levels of ten genes in Pseudomonas sp. M18G proved that this method is universal and feasible for mRNA quantification, and has great potential for application in mRNA quantification in various organisms. [Copyright &y& Elsevier]

Abstract: Near Infrared Chemical Imaging (NIR-CI) is an attractive technique in pharmaceutical development and manufacturing, where new and more robust methods for assessment of the quality of the final dosage products are continuously demanded. The pharmaceutical manufacturing process of tablets is usually composed by several unit operations such as blending, granulation, compression, etc. Having reliable, robust and timely information about the development of the process is mandatory to assure the quality of the final product. One of the main advantages of NIR-CI is the capability of recording a great amount of spectral information in short time. To extract the relevant information from NIR-CI images, several Chemometric methods, like Partial Least Squares Regression, have been demonstrated to be powerful tools. Nevertheless, these methods require a calibration phase. Developing new methods that do not need any prior calibration would be a welcome development. In this context, we study the potential usefulness of Classical Least Squares and Multivariate Curve Resolution models to provide quantitative and spatial information of all the ingredients in a complex tablet matrix composed of five components without the development of any previous calibration model. The distribution of the analytes in the surfaces, as well as the quantitative determination of the five components is studied and tested. [Copyright &y& Elsevier]

Abstract: The development of a fast, reliable and inexpensive protocol for the concentration of bacteria from food by the removal of fat, carbohydrates and proteins that is compatible with downstream alternative DNA-based quantification methods is described. The protocol was used for dairy products, cooked and smoked fish and meat, carbohydrate-rich cooked products, ready-to-eat sauces, egg and blood. Lysis resulted in pellets of reasonable size for further processing. Starch, plant materials, fungi, tissues such as sinew, and chalaza could not be dissolved. Using L. monocytogenes, S. aureus and B. cereus as model organisms, microscopic analysis of the remaining bacterial pellets revealed that the recovered bacteria remained physically intact, albeit that the viability of the cells was compromised. Using real-time PCR, 7.3 CFU of L. monocytogenes were detected in artificially contaminated ultra-high temperature treated (UHT) milk and raw milk. [Copyright &y& Elsevier]

We describe the preparation and characterization of functionalized nano-PbS. The functionalized nanoparticles are water-soluble. Reaction of mercaptoacetic acid functionalized nano-PbS with γ-globulin (γ-IgG) results in an enhanced resonance light scattering (RLS) at 385 nm. Based on this, a new direct quantitative method for γ-globulin in blood serum samples without separation of human serum albumin is established. Under optimal conditions, the enhanced RLS intensity is in proportion to the γ-IgG concentration in the range 10–500 ng ml−1. The 3σ limit of detection is 2.75 ng ml−1. The contents of γ-IgG in blood serum samples were determined with a recovery of 97–104% and R.S.D. of 1.5–2.1% (n=6). This method proved to be very sensitive, rapid, simple and tolerant of most interfering substances. [Copyright &y& Elsevier]

A direct on sampling filter in solution (DOSIS) method for quantification of airborne cat allergens has been developed. In this method, the allergens firmly adsorbed to a porous polytetrafluoroethylene filter are reacted with specific antibodies conjugated to alkaline phosphatase, generating a matrix-bound allergen–antibody–phosphatase complex. The treated filter is subsequently floated on a commercially available chemiluminescent phosphatase substrate solution. Aliquots of this solution are removed and analyzed luminometrically. The light intensity of the product is linearly related to the amount of allergen over a large mass range, 0–100 SQ units (1 SQ unit is about 146 pg of the allergenic protein Fel d 1). DOSIS demonstrated intra- and interassay precisions of 9% and 8% and 14% and 21% for the levels 4 and 20 SQ units per filter, respectively. The limit of quantification was estimated to 0.4 SQ units (58 pg Fel d 1) of cat allergen per filter. Application of DOSIS to analysis of cat allergen concentrations of indoor air in homes with and without cats revealed, on average, a six times higher concentration in the former (142 SQ units/m3) as compared to the latter (24 SQ units/m3). The recorded concentrations for airborne cat allergen in homes with cats are in accordance with previously reported figures. Allergen-specifically stained sampling filters revealed the particulate nature of airborne cat allergen which seemed predominantly to be carried by numerous large dust particles. [Copyright &y& Elsevier]

Reverse transcription quantitative PCR (RT-qPCR) has emerged as the gold standard for virus detection and quantification, being utilized in numerous diagnostic and research applications. However, the direct detection of viruses has so far posed a challenge as the viral genome is often encapsidated by a proteinaceous layer surrounded by a lipid envelope. This necessitates an additional and undesired RNA extraction step prior to RT-qPCR amplification. To circumvent this limitation, we have developed a direct RT-qPCR method for the detection of RNA viruses. In our method, we provide a proof-of-concept using phage phi6, a safe-to-use proxy for pathogenic enveloped RNA viruses that is commonly utilized in e.g. aerosolization studies. First, the phage phi6 envelope is removed by 1% chloroform treatment and the virus is then directly quantified by RT-qPCR. To identify false negative results, firefly luciferase is included as a synthetic external control. Thanks to the duplex format, our direct RT-qPCR method reduces the reagents needed and provides an easy to implement and broadly applicable, fast, and cost-effective tool for the quantitative analysis of enveloped RNA viruses.•One-step direct RT-qPCR quantification of phage phi6 virus without prior RNA isolation.•Reduced reaction volume for sustainable and cost-effective analysis., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s). Published by Elsevier B.V.)

Background It is well known that the difference between prescribed and delivered dialysis doses greatly affects the morbidity and mortality of dialysed patients. The on-line monitoring of delivered dialysis is therefore of paramount importance. Recently, a conductivity- based method for determining Kt/V on a routine basis has been proposed. Methods The study was performed using a specially designed module (Biofeedback Module, COT, Hospal) which, when connected to a dialysis monitor, automatically determines effective ionic dialysance (ID). During three consecutive dialysis sessions, administered to each of eight patients at the same depurative efficiency, we determined Kt/V by using mean effective ionic dialysance and by assuming, as suggested, that urea distribution volume corresponded to 55% of body weight. This method was compared with the gold standard of the direct quantification method. The Kt/V was also calculated by using mean effective ionic dialysance and the volume of urea distribution derived from anthropometric parameters. Result The Kt/V determined by using mean effective ionic dialysance and by assuming that urea distribution volume corresponded to 55% of body weight was heavily underestimated (−22%). This difference was due to both the overestimate of urea distribution volume (+l7%) and underestimate of effective urea clearance (Kueff) (−11%). The mean Kt/V calculated on the basis of ionic dialysance and anthropometric volume was also underestimated (−23%) since this volume was overestimated (+17%). Nevertheless, ionic dialysance and urea clearance proved to be closely correlated (r2=0.89) so that effective urea clearance can be derived according to: Kueff=ID×0.865+39.89. Conclusions In steady-state patients, once urea distribution volume has been correctly determined by means of direct quantification, effective urea clearance can be easily derived from ionic dialysance and Kt/V calculated on-line at each session, without blood sampling or any additional costs. [ABSTRACT FROM PUBLISHER]

FIM protein, which consists of 155 amino acids, was developed as a novel GLP‐1 analog to reduce blood glucose, and pharmacodynamic results showed that it had a certain effect when used in treating Alzheimer's disease. The molecular weight of FIM is 16,304 Da. In theory, the concentration of FIM in biological samples should be determined by the ligand binding assay method or indirectly quantified using LC–MS/MS instrumentation. However, the above methods are complex and time‐consuming. In this study, we successfully developed a simpler LC–MS/MS method for directly quantifying the intact FIM protein in monkey plasma for the first time. The chromatographic separation of FIM was achieved using an InertSustain Bio C18 column with a mobile phase of acetonitrile containing 0.1% formic acid (A)–water containing 0.1% formic acid (B) at a flow rate of 0.3 ml/min. Good linearity was observed in the concentration range of 5–500 ng/ml (r2 > 0.99). The intra‐ and inter‐day precisions (expressed as relative standard deviation, RSD) of FIM were 2.30–12.8 and 7.30–13.2%, respectively. The intra‐ and inter‐day accuracies (expressed as a relative error, RE) were −12.7–6.55 and − 10.1–0.892%, respectively. This method was successfully applied for a pharmacokinetic study of the FIM protein in four monkeys after subcutaneous administration. [ABSTRACT FROM AUTHOR]

• Direct and indirect quantification of phosphate metabolites of nucleoside analogs. • Quantification of intracellular and total tissue concentrations of nucleotide metabolites. • Compare methods for sample collection, preparation, storage conditions, and LC–MS analysis. Nucleoside reverse transcriptase inhibitors (NRTIs) are prodrugs that require intracellular phosphorylation to active triphosphate nucleotide metabolites (NMs) for their pharmacological activity. However, monitoring these pharmacologically active NMs is challenging due to their instability, high hydrophilicity, and their low concentrations in blood and tissues. Liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) is the gold standard technique for the quantification of NRTIs and their phosphorylated NMs. In this review, an overview of the publications describing the quantitative analysis of intracellular and total tissue concentration of NMs is presented. The focus of this review is the comparison of the different approaches and challenges associated with sample collection, tissue homogenization, cell lysis, cell counting, analyte extraction, sample storage conditions, and LC–MS analysis. Quantification methods of NMs via LC–MS can be categorized into direct and indirect methods. In the direct LC–MS methods, chromatographic retention of the NMs is accomplished by ion-exchange (IEX), ion-pairing (IP), hydrophilic interaction (HILIC), porous graphitic carbon (PGC) chromatography, or capillary electrophoresis (CE). In indirect methods, parent nucleosides are 1st generated from the dephosphorylation of NMs during sample preparation and are then quantified by reverse phase LC–MS as surrogates for their corresponding NMs. Both approaches have advantages and disadvantages associated with them, which are discussed in this review. [ABSTRACT FROM AUTHOR]

In this paper, we present a compact Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS) designed for real time analysis of volatile organic compounds (VOCs) in air or in water. The spectrometer is based on a structured permanent magnet made with NdFeB segments. Chemical ionization is implemented inside the ICR cell. The most widely used reaction is the proton transfer reaction using H3O+ precursor ions, but other ionic precursors can be used to extend the range of species that can be detected. Complex mixtures are studied by switching automatically from one precursor to another. The accuracy obtained on the mass to charge ratio (Δm/z 5 × 10−3), allows a precise identification of the VOCs present and the limit of detection is 200 ppb without accumulation. The time resolution is a few seconds, mainly limited by the time necessary to come back to background pressure after the gas pulses. The real time measurement will be illustrated by the monitoring of VOCs produced during the thermal degradation of a polymer and by an example where three different precursor ions are used alternatively to monitor a gas sample. [ABSTRACT FROM AUTHOR]

Being the forensic workflow a long process, with many steps associated, it is highly desirable the development of a method that will allow to select more quickly samples with enough DNA quantity to proceed to amplification. A comparative study was performed to evaluate the viability of a direct DNA quantification methodology in the forensic workflow. [ABSTRACT FROM AUTHOR]

Circulating miRNAs are protected from ribonuclease degradation by assembly into microvesicles and exosomes. Releasing miRNAs completely from these particles is the key step to quantify the circulating miRNAs. Currently purified RNA-based quantitative analysis is widely used while it is time and cost consuming with high risk for those circulating miRNAs with low abundance due to partial loss of RNA during the steps of total RNA extraction and small RNA enrichment. Herein, we optimized a simple, effective and time-saving method to directly measure plasma miRNAs without RNA isolation. It is based on complete miRNA release from the protein complexes, followed by miRNA-specific reverse transcription and quantitative real-time PCR amplification. By comparison to the RNA-based approach, the direct quantification method showed more efficiency for circulating miRNA analysis, higher accuracy and specificity. By application of the direct quantification method to clinical samples combined with the RNA-based miRNA screening analysis, upregulation of miR-106a in blood was validated in metastatic breast cancer patients, indicating miR-106a are a potential biomarker for metastatic breast cancer., Competing Interests: None.