Quantification Technology Research Articles

Development of an innovative technology to segment luminal borders of intravascular ultrasound image sequences in a fully automated manner

Although intravascular ultrasound (IVUS) is the commonest intravascular imaging modality, it still is inefficient for clinical use as it requires laborious manual analysis. This study demonstrates the feasibility of a near real-time fully automated technology for accurate identification, detection, and quantification of luminal borders in intravascular images. This technology uses a combination of the novel approaches of a self-tuning engine, dynamic and static masking systems, radar-wise scan, and contour correction cycle method. The performance of the computer algorithm developed based on this technology was tested on a sequence of IVUS and True Vessel Characterization (TVC) images obtained from the left anterior descending (LAD) artery of 6 patients with coronary artery disease. The accuracy of the algorithm was evaluated by comparing luminal borders traced manually with those detected automatically. The processing time of the developed algorithm was also tested on a Dell laptop with an Intel Core i7-8750H Processor (4.1 GHz with 6 cores, 9 MB Cache). Linear regression and Bland-Altman analyses indicated high correlation between manual and automatic tracings (Y = 0.80 × X+1.70, R2 = 0.88 & 0.67 ± 1.31 (bias±SD)). Whereas analysis of 2000 IVUS images using one CPU core with a 30% load took 23.12 min, the same analysis using six CPU cores with 90% load took 1.0 min. The performance, accuracy, and speed of the presented state-of-the-art technology demonstrates its capacity for use in clinical settings.

Analysis of chlorination by‐products in swimming pool water by membrane introduction mass spectrometry – Influence of water physicochemical parameters

RationaleChlorine reacts in swimming pools with several compounds released by bathers to form disinfection by‐products (DBPs). Epidemiological studies have shown adverse effects on health associated with the exposure to DBPs present in indoor swimming pool atmosphere. DBPs analyses require the use of multiple techniques depending on the targeted molecules. The measurement process itself is challenging due to the low stability of several compounds and the lack of specificity of certain methods. The Membrane Introduction Mass Spectrometry (MIMS) technique provides a solution to these problems by specific and sensitive in situ measurement of DBPs. This study investigates the effect of analytical conditions on quantification of DBPs and assesses the relevance of using MIMS for reliable analysis under typical swimming pool operating conditions.MethodsMIMS is based on the simultaneous permeation of the selected compounds from the air or water samples through a polydimethylsiloxane (PDMS) membrane. DBPs are identified and quantified with a quadrupole analyzer after electron ionization. Limits of quantification (LOQs) of five DBPs are determined to assess the sensitivity of the system. Moreover, signal changes are monitored while varying physicochemical parameters such as temperature, pH and ionic strength.ResultsThe mass spectra obtained for individual molecules show that the simultaneous measurement of trihalomethanes (THMs) and chloramines requires the monitoring of several ions and mathematical corrections of the signal. The pH and ionic strength of the solution do not significantly influence the determination of THMs. On the contrary, the temperature and hydraulics at the membrane interface must be controlled for accurate determination of DBPs.ConclusionsResults confirm that MIMS is a promising technology for the simultaneous quantification of volatile DBPs in both water and air of swimming pools. However, operating conditions such as membrane temperature should be treated with great care in order to avoid interferences.

MicroRNAs: Key Regulators to Understand Osteoclast Differentiation?

MicroRNAs (miRNAs) are small non-coding single-stranded RNAs that represent important posttranscriptional regulators of protein-encoding genes. In particular, miRNAs play key roles in regulating cellular processes such as proliferation, migration, and cell differentiation. Recently, miRNAs emerged as critical regulators of osteoclasts (OCs) biology and have been involved in OCs pathogenic role in several disorders. OCs are multinucleated cells generated from myeloid precursors in the bone marrow, specialized in bone resorption. While there is a growing number of information on the cytokines and signaling pathways that are critical to control the differentiation of osteoclast precursors (OCPs) into mature OCs, the connection between OC differentiation steps and miRNAs is less well-understood. The present review will first summarize our current understanding of the miRNA-regulated pathways in the sequential steps required for OC formation, from the motility and migration of OCPs to the cell-cell fusion and the final formation of the actin ring and ruffled border in the functionally resorbing multinucleated OCs. Then, considering the difficulty of working on primary OCs and on the generation of robust data we will give an update on the most recent advances in the detection technologies for miRNAs quantification and how these are of particular interest for the understanding of OC biology and their use as potential biomarkers.

Transcriptome and Proteome-Based Network Analysis Reveals a Model of Gene Activation in Wheat Resistance to Stripe Rust.

Stripe rust, caused by the pathogen Puccinia striiformis f. sp. tritici (Pst), is an important fungal foliar disease of wheat (Triticum aestivum). To study the mechanism underlying the defense of wheat to Pst, we used the next-generation sequencing and isobaric tags for relative and absolute quantification (iTRAQ) technologies to generate transcriptomic and proteomic profiles of seedling leaves at different stages under conditions of pathogen stress. By conducting comparative proteomic analysis using iTRAQ, we identified 2050, 2190, and 2258 differentially accumulated protein species at 24, 48, and 72 h post-inoculation (hpi). Using pairwise comparisons and weighted gene co-expression network analysis (WGCNA) of the transcriptome, we identified a stress stage-specific module enriching in transcription regulator genes. The homologs of several regulators, including splicing and transcription factors, were similarly identified as hub genes operating in the Pst-induced response network. Moreover, the Hsp70 protein were predicted as a key point in protein–protein interaction (PPI) networks from STRING database. Taking the genetics resistance gene locus into consideration, we identified 32 induced proteins in chromosome 1BS as potential candidates involved in Pst resistance. This study indicated that the transcriptional regulation model plays an important role in activating resistance-related genes in wheat responding to Pst stress.

Self-emulsifying drug delivery system: Mucus permeation and innovative quantification technologies.

Mucus is a dynamic barrier which covers and protects the underlying mucosal epithelial membrane against bacteria and foreign particles. This protection mechanism extends to include therapeutic macromolecules and nanoparticles (NPs) through trapping of these particles. Mucus is not only a physical barrier that limiting particles movements based on their sizes but it selectively binds with particles through both hydrophilic and lipophilic interactions. Therefore, nano-carriers for mucosal delivery should be designed to eliminate entrapment by the mucus barrier. For this reason, different strategies have been approached for both solid nano-carriers and liquid core nano-carriers to synthesise muco-diffusive nano-carrier. Among these nano-strategies, Self-Emulsifying Drug Delivery System (SEDDS) was recognised as very promising nano-carrier for mucus delivery. The system was introduced to enhance the dissolution and bioavailability of orally administered insoluble drugs. SEDDS has shown high stability against intestinal enzymatic activity and more importantly, relatively rapid permeation characteristics across mucus barrier. The high diffusivity of SEDDS has been tested using various in vitro measurement techniques including both bulk and individual measurement of droplets diffusion within mucus. The selection and processing of an optimum in vitro technique is of great importance to avoid misinterpretation of the diffusivity of SEDDS through mucus barrier. In conclusion, SEDDS is a system with high capacity to diffuse through intestinal mucus even though this system has not been studied to the same extent as solid nano-carriers.

Image quantification technology of the heterochromatin and euchromatin region for differential diagnosis in the lobular endocervical glandular hyperplasia.

Lobular endocervical glandular hyperplasia (LEGH) was first described by Nucci et al. in 1999 and is believed to be a precancerous lesion of minimal deviation adenocarcinoma and gastric-type adenocarcinoma in the uterine cervix. LEGH lesions do not always exhibit apparent cellular and structural atypia, so are difficult to distinguish from normal endocervical cells (EC cells) with cytological examination. Therefore, we often struggle to make a definite diagnosis of LEGH. We used microscopy images of cytological specimens that were diagnosed as EC cells and LEGH cells. Signal intensity in whole nuclear area and in heterochromatin and euchromatin regions, euchromatin area ratio, and nuclear morphological features were quantified in each cell nucleus of the cases. Statistical analyses were conducted to determine statistical significance. Finally, we performed linear support vector machine (LSVM) modeling as a discriminant analysis using the quantified features. Signal intensity in whole nuclear area, and heterochromatin and euchromatin regions of EC cell nuclei were higher than that of the LEGH cell nuclei. Morphologically, EC cell nuclei were larger than LEGH cell nuclei, and nuclei of LEGH cells had irregular nuclear respectively membrane structure and an elongated shape. The LSVM accuracy of 10-fold cross validation and leave-one-case-out cross-validation (LOCOCV) using all measured features were 84.7% to 89.3% and 78.6% to 86.0%, respectively. The LVSM analysis using features extracted from signal intensity and morphological analysis was useful for discrimination of EC cells vs LEGH cells. We therefore believe that this image analysis method could be used for early detection of LEGH.

Droplet Digital PCR: A New Technology for Detection and Quantification of Chimerism After Allogenic Hematopoietic Stem Cell Transplantation

Droplet Digital PCR: A New Technology for Detection and Quantification of Chimerism After Allogenic Hematopoietic Stem Cell Transplantation

Comparison between the performance of Two-Dimensional and Point Shear Wave elastography for the noninvasive assessment of liver cirrhosis

Background and aim The goals of this study were to compare the noninvasive diagnostic performance of two elastography techniques for the diagnosis of liver cirrhosis in a cohort of patients with known hepatitis C virus using Transient Elastography FibroScan as the method of reference since it is a validated tool for the non-invasive assessment of liver fibrosis. Material and Methods The study included 122 patients aged 38-80 (62 ± 7.9), 35% male (43/122), 65% female (79/122) with known hepatitis C virus in whom liver stiffness was evaluated during the same session by means of three elastography Methods: Point Shear Wave Elastography (pSWE) using virtual touch quantification(VTQ) technology Siemens Acuson S2000TM, Two-Dimensional Shear Wave Elastography (2D-SWE) embedded in General Electrics LOGIQ E9 ultrasound machine and Transient Elastography(TE;FibroScan, EchoSens). Reliable LS measurements were defined, for all techniques, as the median value of 10 measurements with an interquartile range/median(IQR/MED) Results Reliable liver stiffness measurements were obtained in 93% (114/122) of cases by means of TE, 88% (107/122) of cases by means of VTQ and 87% (106/122) of cases by means of 2D-SWE.GE. In the final analysis we included 93 patients which had reliable stiffness measurements with all methods.The AUROCS were calculated considering TE as the reference method. The optimal cutoff of VTQ for cirrhosis was 1.77m/s(SE:83%; SP:88%; AUC:0.89) and for 2D-SWE.GE was 10.2 kPa(SE:86%; SP:77%; AUC:0.89). No statistical differences were found between both methods(p Conclusion Both methods have similar performance for diagnosing liver cirrhosis and seem to be a promising alternative to Transient Elastography in evaluating patients with liver fibrosis.

Current Trends in Detection of Histamine in Food and Beverages.

Histamine is a heterocyclic amine formed by decarboxylation of the amino acid l-histidine. It is involved in the local regulation of physiological processes but also can occur exogenously in the food supply. Histamine is toxic at high intakes; therefore, determination of the histamine level in food is an important aspect of food safety. This article will review the current understanding of physiological functions of endogenous and ingested histamine with a particular focus placed on existing and emerging technologies for histamine quantification in food. Methods reported in this article are sequentially arranged and provide a brief overview of analytical methods reported, including those based on nanotechnologies.

Accounting for equality: Gender budgeting and moderate feminism

This article sheds light on the relationship between quantification and calculative practices of accounting and moderate feminism. Drawing on a case of gender budgeting in Austria and the literature on social studies of accounting, I show how gender equality initiatives are translated into practice and come to codify the governance of gender relations through calculative practices that further the logic of neoliberal governmentality, rather than fundamentally challenging it. As such, this article provides an account of one site of the neoliberal recuperation of feminist critique through technologies of quantification and accountability.

Seropositivity and Antibody Profiling of Patients Are Dramatically Impacted by the Features of Peptides Used as Immunosorbents: A Lesson from Anti-Citrullinated Protein/Peptide Antibody.

Quantification of Abs toward a single epitope is critical to understanding immunobiological processes. In autoimmunity, the prognostic value of the serological profiles of patients draws much attention, but the detection of Abs toward a single epitope is not well controlled. Particularly, the rheumatoid arthritis (RA)-specific anti-citrullinated protein/peptide Abs (ACPA) are specific to a two-atom change on arginyl residues and are considered a heterogeneous family of Abs. As a model, we studied ACPA to decipher how peptide features used as immunosorbent impact Ab detection. We synthesized 30 peptides encompassing immunodominant epitopes of citrullinated fibrin differing by their length and biotin location and tested them using ELISA with 120 sera from RA and non-RA rheumatic disease controls, generating over 3000 experimental measurements. We showed that minor molecular changes in peptide chemical structure had dramatic consequences. Even when peptides exhibited the same epitope, measured Ab titers were extremely variable, and patients' seropositivity was discordant in up to 50% of cases. The distance between epitope and biotin was the most critical parameter for efficient Ab detection irrespective of biotin position or peptide length. Finally, we identified a 15-mer peptide bearing a single citrullinated epitope detecting almost all ACPA-positive sera, thus revealing a high degree of homogeneity in RA autoimmune response. This integrative analysis deciphers the dramatic impact of the molecular design of peptide-based technologies for epitope-specific Ab quantification. It provides a model for assay development and highlights that the studies using such technologies can give a wrong perception of biological processes and therefore that medical use of data must be cautious.

Effects of smoking on the lower respiratory tract microbiome in mice

BackgroundRecent studies break with traditional opinion that the lower respiratory tract is sterile, and increasingly focus on the lung microbiome and disease. Smoking, as an important etiology of inflammatory lung disease, was considered as a factor influencing lung microbiome variations in our study, and we aimed to study the effect of smoking on inflammation and microbial diversity and community.MethodsForty male mice were selected and randomly divided into a smoking and a non-smoking group. Mice in the smoking group were exposed to smoke smog for 2 h/day for 90 days. Blood and lung tissues were obtained after the experiment, and ELISA was used to measure interleukin-6 and C reactive protein concentrations. 16S rRNA gene quantification and sequencing technology were used to compare microbial diversity and community between the two groups. SAS 9.1 and R software were used to analyze the data.ResultsThirty-six mice survived, and the weight of the smoking group increased more slowly than that of the non-smoking group. Denser inflammation and congestion were observed in the lungs of the smoking mice compared with the non-smoking group Higher microbial diversity was observed in the smoking group, and Enterobacter, Acidimicrobiales_norank, and Caulobacteraceae_Unclassified genus were significantly more abundant in the non-smoking group (P < 0.001).ConclusionsSmoking altered microbial diversities and communities in the lower respiratory tract of mice. Microbial variation should be considered in future studies focusing on smoking-induced inflammatory disease.

Acceleration of nano-surface and molecular-orientation limited (nSMOL) proteolysis with acidified reduction pretreatment for quantification of Tocilizumab

Antibody drugs are effective therapeutic agents and provide treatment for many types of diseases such as cancer and rheumatoid arthritis. Because many antibody drugs are developed and approved, quantification technologies of antibodies are required for drug development and individualized therapy. We recently reported a high reproducible and robust therapeutic drug monitoring method for antibody drugs. This method was developed to be applicable to all type of antibody drugs and to provide accurate quantification values. The method is named nano-surface and molecular-orientation limited (nSMOL) proteolysis. nSMOL limits the access of protease to immunoglobulin G molecules and aims to selectively proteolyze on Fab region of substrate. However, the bioanalysis of Tocilizumab using nSMOL has not been validated. We newly discovered that acidified reduction pretreatment of Tocilizumab promotes digestive efficiency by nSMOL proteolysis. Exposure of Tocilizumab to Tris(2-carboxyethyl)phosphine hydrochloride before nSMOL proteolysis significantly improved the recovery of peptides. Under this condition, the quantification range of Tocilizumab in human serum was from 0.781 to 200 μg/mL. The quantification values of quality control samples fulfilled all guideline criteria for bioanalytical validation. The signature peptide with the highest quantitative sensitivity was on H-chain VTMLR. From these results, it is expected that the pretreatment with TCEP will broaden the application of antibody drugs quantification by nSMOL proteolysis.

Gold nanoparticles - an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis.

PurposeThe objective of this study is to develop a facile tool for the absolute detection and quantification of nucleic acid transcripts, using a gold nanoparticle-based optical biosensor. Topoisomerase 1 (TOP1) and tyrosyl DNA phosphodiesterase 2 (TDP2) were among the nucleic acid transcripts of choice due to their role as genomic instability biomarkers and their implication in various cancers and neurologic disorders. This opens the door to develop a simple tool that can be used for diagnosing and monitoring treatment response for such diseases, overcoming the requirements for high cost, time, and complexity of the existing technologies for the absolute quantification of transcripts of interest.Materials and methodsThe TOP1 and TDP2 mRNA transcripts were first captured specifically using magnetic nanoparticles that were functionalized with TOP1- and TDP2-specific probes, respectively. The captured mRNA was then directly detected and quantified using the gold aggregating gold (GAG) assay, without the need for amplification as in existing technologies used for the quantification of transcripts.ResultsA linear correlation exists between the GAG assay and the qPCR for the quantification of the TOP1 and TDP2 mRNA transcripts (101–104 copies). The detection limit of the GAG assay in mRNA quantification was up to 10 copies per reaction. Wild-type and TDP2-deficient cell lines confirmed the assay specificity and reproducibility in distinguishing between different transcripts.ConclusionThe GAG assay can be utilized as an inexpensive, rapid, simple, and sensitive tool for the absolute quantification of RNA for different applications, instead of the laborious, expensive, and sophisticated real-time PCR.

Autonomous system for rapid field quantification of Escherichia coli in surface waters.

The purpose of this work is to present and evaluate the performance of a novel Automatic Lab-in-vial Escherichia coli Remote Tracking technology based on an automated real-time defined substrate approach, implemented in both portable and insitu instruments. We present the fresh water calibration procedure, and assess performance using side-by-side comparison with most probable number (MPN) approaches in terms of accuracy, reproducibility and capability to correctly generate early-warning alerts. Long-term data from an operational insitu deployment at a potential bathing site is presented as well. Automatic Lab-in-vial Escherichia coli Remote Tracking technology is shown to be an accurate and rapid bacterial quantification technology, capable of autonomous insitu measurements with metrological capabilities comparable to those of an approved laboratory using MPN microplate techniques. Rapid quantification of bacterial pollution is a requirement in water quality applications ranging from recreational water use, agriculture and aquaculture to drinking and wastewater treatment. The method and instruments presented in this work should enable fast and accurate bacterial concentration measurements to be performed in a portable or insitu manner, thus simplifying operational logistics, reducing time-to-result delays, and eliminating sample transportation constraints associated with traditional techniques.

非住宅建築物を対象とした先導的省エネルギー技術の評価方法に関する研究 その6 外気冷房および換気量制御に関するエネルギー削減量に関する検討

非住宅建築物を対象とした先導的省エネルギー技術の評価方法に関する研究 その6 外気冷房および換気量制御に関するエネルギー削減量に関する検討

Absolute quantification of the α, α′, and β subunits of β-conglycinin from soybeans by liquid chromatography/tandem mass spectrometry using stable isotope-labelled peptides

β-Conglycinin, a major protein in soybeans, shows improvement effect of lipid metabolism. Moreover, this protein influences the processing properties of soybeans. β-Conglycinin is a hetero-trimer constituted by α, α′, and β subunits. In this work, a method for the selective quantification of these subunits was developed by means of protein absolute quantification (AQUA) technology using liquid chromatography/tandem mass spectrometry with the stable isotope-labelled internal standard peptides LQSGDALR[13C6,15N4], NILEASYDTK[13C6,15N2], and NPIYSNNFGK[13C6,15N2]. This method exhibited linear relationships (r2 > 0.99) in the concentration range of 1.2–300 fmol/μL for LQSGDALR[13C6,15N4] and NILEASYDTK[13C6,15N2], and of 4.7–300 fmol/μL for NPIYSNNFGK[13C6,15N2]. As a result, the content of these subunits in β-conglycinin-rich and both α and α′ subunit-deficient soybean cultivars was successfully determined. This quantitative assay is promising for the evaluation of the food functionality and processing properties of soybeans.

Molecular Subtypes of Urothelial Bladder Cancer: Results from a Meta-cohort Analysis of 2411 Tumors

BackgroundPrevious molecular subtyping for bladder carcinoma (BLCA) involved <450 samples, with diverse classifications. ObjectiveTo identify molecular subtypes by curating a large BLCA dataset. Design, setting, and participantsGene expression publicly available were combined and reanalyzed. The dataset contained 2411 unique tumors encompassing non-muscle-invasive (NMIBC) and muscle-invasive BLCA (MIBC). Subtypes were reproduced on The Cancer Genome Atlas, UROMOL, and IMvigor210. InterventionSubtypes were assigned by gene expression. Outcome measurements and statistical analysisKaplan-Meier analyses were performed for subtype-clinical outcome correlations; Chi-square/Fisher exact tests were used for subtype-clinicopathological parameters associations. Results and limitationsWe identified six molecular subtypes with different overall survival (OS) and molecular features. Subtype Neural-like (median OS, 87 mo) is prevalent in MIBC and characterized by high WNT/β-catenin signaling. HER2-like (107.7 mo) is distributed evenly across NMIBC and MIBC, with higher ERBB2 amplification and signaling. Papillary-like (>135 mo), an NMIBC subtype enriched in urothelial differentiation genes, shows a high frequency of actionable FGFR3 mutations, amplifications, and FGFR3-TACC3 fusion. Luminal-like (91.7 mo), predominantly NMIBC, has higher MAPK signaling and more KRAS and KMT2C/D mutations than other subtypes. Mesenchymal-like (MES; 86.6 mo) and Squamous-cell carcinoma-like (SCC; 20.6 mo) are predominant in MIBC. MES is high in AXL signaling, whereas SCC has elevated PD1, CTLA4 signaling, and macrophage M2 infiltration. About 20% of NMIBCs show MIBC subtype traits and a lower 5-yr OS rate than Papillary-like NMIBC (81% vs 96%). The main limitations of our study are the incomplete clinical annotation, and the analyses were based on transcriptome subset due to comparisons across gene expression quantification technologies. ConclusionsBLCA can be stratified into six molecular subtypes. NMIBC, with a high risk of progression, displays the molecular features of MIBC. Patient summaryBiomarkers are urgently needed to guide patient treatment selection and avoid unnecessary toxicities in those who fail to respond. We believe molecular subtyping is a promising way to tailor disease management for those who will benefit most.

Glyphosate detection: methods, needs and challenges

Glyphosate is considered toxicologically harmful and presents potential association with human carcinogenesis and other chronic diseases, including mental and reproductive behaviors. The challenges to analyse and demonstrate its toxicity are likely due to its metal-chelating properties, the interference of organic compounds in the environment, and similarity with its by-products. Whereas there is a link with serious health and environmental problems, there is an absence of public health policies, which is probably due to the difficulties in detecting glyphosate in the environment, further complicated by the undetectable hazard in occupational safety and health. The historical lenient use of glyphosate in transgenic-resistant crops, corroborated by the fact that it is not easily detected, creates the “Glyphosate paradox”, by which it is the most widely used herbicide and one of the most hardly determined. In this review, we revisited all available technologies for detection and quantification of glyphosate, including their drawbacks and advantages, and we further discuss the needs and challenges. Briefly, most of the technologies require high-end equipments and resources in low throughput, and none of them are adequate for real-time field tests, which may explain the lack of studies on occupational health associated with the chemical hazard. The real-time detection is an urgent and highly demanded need to improve public policies.

Quantitative analysis of amino acids in human and bovine colostrum milk samples through iTRAQ labeling

The types and quantity of proteins vary widely between bovine and human milk, with corresponding differences in free and hydrolytic amino acids. In this study, the free and hydrolytic amino acids of bovine and human colostrum were for the first time qualitatively and quantitatively determined using isobaric tags for relative and absolute quantification technology combined with liquid chromatography tandem mass spectrometry detection. Total free amino acid content was 0.32 g L-1 and 0.63 g L-1 in bovine and human colostrum respectively, with free amino acid content in human colostrum twice that of bovine colostrum. However, total hydrolytic amino acid content was 4.2 g L-1 and 2.2 g L-1 in bovine and human colostrum respectively. We found that the hydrolytic amino acid content in bovine colostrum was higher than that in human colostrum; however, the amount of free amino acids and the overall amino acid content in human colostrum were respectively substantially higher and more varied than in bovine colostrum. Our findings revealed differences between bovine and human colostrum, with these data providing the basis for further research into amino acid metabolomics and infant formula. © 2018 Society of Chemical Industry.