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The present work proposes the development of a simple and easy-to-make microfluidic paper-based analytical device (µPAD) as a prototype chromatic device for the simultaneous determination of copper and zinc in urine. The µPAD comprises a main circular sampling zone with two identical arms, each containing a circular pretreatment zone and a circular detection zone. Copper(II) and zinc(II) react with 1-(2-pyridylazo)-2-naphthol (PAN) reagent, at pH 6.0 and room temperature, to form chelates colored red and pink, respectively, that can be quantified by digital imaging analysis. The colorimetric reaction sensitivity significantly improved by coating the paper substrate surface with 5% 3-aminopropyltriethoxysilane (APTES) and applying three successive additions of standard or sample onto the sampling zone, without any prior sample preparation step. These combined analytical strategies increased the color intensity of the metal chelates formed on µPAD surface, allowing to detect the desired elements at trace level concentrations. The reaction interferences associated with the presence of foreign elements were eliminated or reduced through the addition of masking agents at different concentrations for each case. Under optimized conditions, excellent linear relationships were obtained in the range 0.1−10 µg mL−1, with determination coefficients (R2) greater than 0.9957. The limits of detection (LOD) and quantification (LOQ), respectively, were 27.1 µg L−1 and 90.3 µg L−1 for copper and 35.9 µg L−1 and 119.7 µg L−1 for zinc. The developed prototype device was validated by analyzing urine certified material samples, obtaining a good concordance without significant difference between the certified values and the measured values. Its application for clinical analysis offers simple, quick, cheap, noninvasive, and environmentally friendly measurements, which are desirable analytical features for point-of-care testing.

Summary: Objective: To explore the therapeutic effect of systemic administration combined with microwave ablation (MWA) under computed tomography (CT) and fiberoptic bronchoscope for treating lung cancer. Methods: Sixty-six patients with advanced lung cancer admitted to our hospital from February 2019 to February 2020 were collected and divided into control group and experimental group with 33 patients in each group. The control group was treated with systemic administration, and the experimental group was treated with systemic administration combined with MWA under CT and fiberoptic bronchoscope. Overall response rate (ORR), adverse events (AEs) during treatment, and survival analysis were used to evaluate the curative effect of lung cancer treatment in each group. Results: MWA under CT and fiberoptic bronchoscope could safely remove the cancerous tissues by point burning without destroying the adjacent normal tissues with high success rate. The ORR of the control group was 24.4%, and that of the experimental group was 63.6%, which was better than the control group. The AEs occurred during treatment in each group were of level 1 or level 2, and no serious life-threatening AEs occurred. Progression-free survival (PFS) time and overall survival (OS) time in the experimental group were both longer than those in the control group. Patients treated with MWA had a lower risk of disease progression and death than those treated with systemic administration alone. Conclusion: The treatment of lung cancer using systemic administration combined with MWA under CT and fiberoptic bronchoscope is more effective than using systemic administration alone, which can be promoted in clinical treatment.

Ion mobility spectrometry (IMS) is an analytical technique where ions are separated in the gas phase based on their mobility through a buffer gas in the presence of an electric field. An ion passing through an IMS device has a characteristic collisional cross section (CCS) value that depends on the buffer gas used. IMS can be coupled with mass spectrometry (MS), which characterizes an ion based on a mass-to-charge ratio (m/z), to increase analytical specificity and provide further physicochemical information. In particular, IMS-MS is of ever-increasing interest for the analysis of lipids, which can be problematic to accurately identify and quantify in bodily fluids by liquid chromatography (LC) with MS alone due to the presence of isomers, isobars, and structurally similar analogs. IMS provides an additional layer of separation when combined with front-end LC approaches, thereby, enhancing peak capacity and analytical specificity. CCS (and also ion mobility drift time) can be plotted against m/z ion intensity and/or LC retention time in order to generate in-depth molecular profiles of a sample. Utilization of IMS-MS for routine clinical laboratory testing remains relatively unexplored, but areas do exist for potential implementation. A brief update is provided here on lipid analysis using IMS-MS with a perspective on some applications in the clinical laboratory.

Introduction and Objectives: Liver injury in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant- and Omicron subvariant-infected patients is unknown at present, and the aim of this study is to summarize liver injury in these patients. Patients and Methods: In this study, 460 SARS-CoV-2-infected patients were enrolled. Five severe or critical patients were excluded, and 34 patients were also excluded because liver injury was not considered to be related to SARS-CoV-2 infection. Liver injury was compared between Omicron and non-Omicron variants- and between Omicron subvariant-infected patients; additionally, the clinical data related to liver injury were also analyzed. Results: Among the 421 patients enrolled for analysis, liver injury was detected in 76 (18.1%) patients, including 46 Omicron and 30 non-Omicron variant-infected patients. The ratios did not differ between Omicron and non-Omicron variant-, Omicron BA.1, BA.2 and BA.5 subvariant-infected patients (P>0.05). The majority of abnormal parameters of liver function tests were mildly elevated (1-3 × ULN), the most frequently elevated parameter of liver function test was γ-glutamyl transpeptidase (GGT, 9.5%, 40/421), and patients with cholangiocyte or biliary duct injury markers were higher than with hepatocellular injury markers. Multivariate analysis showed that age (>40 years old, OR=1.898, 95% CI=1.058–3.402, P=0.032), sex (male gender, OR=2.031, 95% CI=1.211–3.408, P=0.007), serum amyloid A (SAA) level (>10 mg/ml, OR=3.595, 95% CI=1.840–7.026, P

As an important biomarker for renal related diseases, detection of urea is playing a vital role in human biofluids on clinical diagnosis concern. In this work, a synthetic salicyaldehyde based imine fluorophore was synthesized using sonication method and conjugated with urease which was used as fluorescent biosensor for the detection of urea in serum samples. This enzyme based biosensor has shown a good selectivity and sensitivity towards urea with the linear range from 2 to 80 mM and the detection limit of 73 µM. The sensing response obtain is highly agreeing with existing analytical technique for urea detection which strongly recommends this biosensor for clinical application., Competing Interests: Declaration of competing interest 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., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In the human body, oxalate tends to form calcium oxalate with calcium ions, which can trigger the formation of stones in the urinary system. Therefore, oxalate in urine is usually utilized as a crucial biomarker in clinical urolithiasis diagnoses. In this work, a turn-on fluorescent nanoprobe was developed based on nitrogen-doped carbon dots (N-CDs) and MnO 2 nanosheets (NSs) nanocomposites for oxalate sensing in urolithiasis patients. MnO 2 NSs is a good sensing platform with high extinction coefficients and rich redox chemistry. The fluorescent N-CDs can be quenched efficiently by MnO 2 NSs through the inner filter effect (IFE) to form N-CDs-MnO 2 nanocomposites. The reductive oxalate could operate the decomposition of MnO 2 NSs to Mn 2+ resulting in the dissociation of the N-CDs-MnO 2 nanocomposites and fluorescence recovery of N-CDs. Under optimal conditions, the developed sensor revealed good specificity toward oxalate with a limit of detection (LOD) of 0.69 μM. The developed sensor was successfully applied to quantify oxalate content in 47 urine samples (41 urolithiasis patients and 6 healthy persons). The results showed great consistency with clinical diagnostic reports and computed tomography images. This novel method retains several unique advantages, including affordable, rapid, and validating potential clinical application., Competing Interests: Declaration of competing interest 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., (Copyright © 2023 Elsevier B.V. All rights reserved.)

It is well known that the coexisting metal ions could significantly influence the atomic spectroscopy (AS) analysis. In this work, a cation-modulated mercury ions (Hg 2+ ) strategy via chemical vapor generation (CVG) was developed for oxalate assay due to the phenomenon that the Ag  +  can significantly reduce the Hg 2+ signal. The regulation effect was studied in depth via experimental investigations. Since Ag  +  can be reduced to silver nanoparticles (Ag NPs) by reductant SnCl 2 , the decrease of the Hg 2+ signal is attributed to the formation of a silver-mercury (Ag-Hg) amalgam. Due to the oxalate can react with Ag  +  to generate Ag 2 C 2 O 4 , which can reduce the generation of Ag-Hg amalgam, a portable and low-power point discharge chemical vapor generation atomic emission spectrometry (PD-CVG-AES) system was constructed to quantify the content of oxalate via monitoring the signal of Hg 2+ . Under optimal conditions, the limit of detection (LOD) was as low as 40 nM in the range of 0.1-10 μM for oxalate assay, while exhibiting good specificity. This method was applied to quantitative oxalate in 50 clinical urine samples of urinary stones patients. The levels of oxalate detected in clinical samples were consistent with clinical imaging results, which is promising for point-of-care testing in clinical diagnosis., Competing Interests: Declaration of competing interest 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., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Although insulin is an essential medicine and a life-saving drug, it has also been incriminated in many poisoning deaths; accidental, suicidal and some with malicious intent. Overdosing with insulin precipitates a life-threatening state of hypoglycemia and if untreated leads to coma, irreversible brain damage and death. Normally, the pancreatic β-cells secrete equimolar amounts of insulin and C-peptide into the portal venous blood, although under physiological conditions the plasma concentration ratio (insulin/C-peptide) is less than unity, because insulin is more susceptible to hepatic first-pass metabolism. A high ratio of insulin/C-peptide in plasma from a poisoned patient is compelling evidence that pharmaceutical insulin was administered, which does not contain C-peptide. The analysis of insulin and C-peptide was traditionally done by immunoassay methods (RIA and/or ELISA), although high resolution LC-MS/MS is more suitable for forensic purposes and permits the identification of insulin analogues. Use of insulin as a murder weapon is exemplified by the case of Colin Norris, a male nurse found guilty of murdering four elderly patients and the attempted murder of a fifth by injecting them with insulin. However, the prosecution evidence against Norris was mainly circumstantial and hearsay. Toxicological evidence against Norris consisted of a high insulin/C-peptide concentration ratio in plasma from one of the victims. This analysis was done by an immunoassay method at a clinical laboratory and not a forensic laboratory. Analytical procedures, including chain-of-custody routines, are more stringent at forensic laboratories. Since his conviction, some of the medical evidence against Norris has been called into question, especially the prevalence of spontaneous attacks of hypoglycemia in elderly and frail patients with co-morbidities., Competing Interests: Declaration of competing interest Declaration of competing interests., (Copyright © 2023. Published by Elsevier Ltd.)

Routine immunoassays for insulin and C-peptide have the potential to cross-react with partially processed proinsulin products, although in healthy patients these are present at such low levels that the interference is insignificant. Elevated concentrations of proinsulin and des-31,32 proinsulin arising from pathological conditions, or injected insulin analogues, however can cause significant assay interferences, complicating interpretation. Clinical diagnosis and management therefore sometimes require methods that can distinguish true insulin and C-peptide from partially processed proinsulin or injected insulin analogues. In this scenario, the high specificity of mass spectrometric analysis offers potential benefit for patient care. A high throughput targeted LC-MS/MS method was developed as a fit for purpose investigation of insulin, insulin analogues, C-peptide and proinsulin processing intermediates in plasma samples from different patient groups. Using calibration standards and bovine insulin as an internal standard, absolute concentrations of insulin and C-peptide were quantified across a nominal human plasma postprandial range and correlated strongly with immunoassay-based measurements. The ability to distinguish between insulin, insulin analogues and proinsulin intermediates in a single extraction is an improvement over existing immunological based techniques, offering the advantage of exact identification of the species being measured. The method promises to aid in the detection of circulating peptides which have previously been overlooked but may interfere with standard insulin and C-peptide immunoassays., Competing Interests: Declaration of Competing Interest 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., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Contactless conductivity detection (C4D) as a universal detection technique plays an important role in combination with efficient electrophoretic separation carried out in capillaries (CE) or on microchips (ME) in the analysis of clinical samples. C4D is particularly sensitive in the quantification of low molecular weight biogenic substances such as inorganic cations and anions, amino acids, amines, low molecular weight organic acids, saccharides and many drugs such as antibiotics, analgesics, anaesthetics or antiepileptics. Biogenic substances are determined in CE/C4D or ME/C4D directly in their native form without derivatization and sample matrix treatment is often based only on dilution or addition of an organic solvent. The limit of detection for most CE/C4D determinations is at the micromolar concentration level, which is sufficient to monitor physiological or therapeutic levels of most of low molecular weight biogenic substances. Therefore, CE/C4D and ME/C4D are widely used for sequential monitoring of nutrients, metabolites and waste products at the level of individual tissues and organs, low-invasive detection of inborn errors of metabolism and cystic fibrosis, pharmacokinetic monitoring and therapeutic drug monitoring. Innovative trends such as electrophoretic stacking, microdialysis, electromembrane extraction, portable and disposable CE instruments and minimally invasive clinical sampling techniques are mentioned. A critical evaluation of the positives and negatives of this technique is presented, covering the main applications published over the last 10 years., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Sensitive electrophoretic determination of 3-hydroxybutyrate (3HB) as an indicator of human ketogenesis is performed in fused silica capillary covalently coated by an anionic copolymer of poly(acrylamide-co-sodium-2-acrylamido-2-methylpropanesulphonate) (PAMAMPS). Baseline separation of 3HB from other components of human serum is achieved in a 20 μm capillary with an effective length of 17 cm covered by 4% PAMAMPS, which generates a cathodic EOF with a mobility of 8.30 ± 0.00 · 10 -9  m 2 /V.s in 80 mM MES/His as background electrolyte. 3HB migrates in counter-current electrophoretic mode against EOF, that effectively improving electrophoretic resolution. Sample pre-treatment is based on adding of 45 μL acetonitrile to 15 μL serum and, after shaking, a 28 mm long zone of supernatant is injected into the capillary, and sharpened after turning on a separation voltage of 20 kV using the technique of large volume sample stacking, where the EOF forces the residual acetonitrile from the capillary. When combined with universal contactless conductivity detection, the achieved LOD and LOQ are 0.43 μM and 1.44 μM, respectively, that are sufficiently low for monitoring the physiological 3HB level. The performed clinical study subsequently showed that serum 3HB increases from a concentration of 71 μM, corresponding to normal food, to level of 1924 μM after 60 h of fasting and returns to the normal physiological concentration 48 h after commencing consumption of high-saccharide food., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Lipid extraction is a critical step in sample preparation of lipidomics studies. Biphasic liquid-liquid extraction protocol with methyl tert-butyl ether (MTBE)/methanol (MeOH) as organic solvents are widely adopted by researchers nowadays as an eco-friendly replacement of classic Folch, and Bligh&Dyer protocols. Yet, it has some limitations such as suboptimal performance for the most polar lipids (e.g. acylcarnitines), complicated handling as it requires phase separation, and is therefore non-ideal for large-scale clinical studies. To advance the extraction protocol for large-scale clinical lipidomics, in this study we explored i) 6 different extraction solvent systems, ii) distinct processing procedures (sonication, mechanical cell lysis and bead homogenizer), and iii) also 7 different reconstitution solvents. The extraction systems investigated included biphasic systems MTBE/MeOH/H 2 O and Hexane/2-propanol (IPA)/1 M acetic acid, and monophasic systems MTBE/MeOH/CHCl 3 , IPA/H 2 O (90% IPA), MeOH/MTBE/IPA, and IPA/H 2 O/MTBE as solvent system for lipid extraction of human platelets. Extraction recovery was investigated by repeated extraction cycles. Subcellular extraction efficiency was assessed by the mitochondria-specific cardiolipins. It turned out that monophasic extraction with MeOH/MTBE/IPA (1.3:1:1, v/v/v), bead homogenizer for cell disruption and MeOH/MTBE (1:1, v/v) as reconstitution solvent provide optimal cellular and subcellular extraction efficiencies for both polar (e.g. acylcarnitines) and apolar lipids (e.g. triglycerides). It is simplified (no phase separation), eco-friendly (reduced solvent consumption and no halogenated ones), fast (5 min for 24 samples in parallel), and can be easily adapted for cells, plasma, and tissue. Therefore, it has great potential for large-scale clinical lipidomics studies., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: To explore the therapeutic effect of systemic administration combined with microwave ablation (MWA) under computed tomography (CT) and fiberoptic bronchoscope for treating lung cancer., Methods: Sixty-six patients with advanced lung cancer admitted to our hospital from February 2019 to February 2020 were collected and divided into control group and experimental group with 33 patients in each group. The control group was treated with systemic administration, and the experimental group was treated with systemic administration combined with MWA under CT and fiberoptic bronchoscope. Overall response rate (ORR), adverse events (AEs) during treatment, and survival analysis were used to evaluate the curative effect of lung cancer treatment in each group., Results: MWA under CT and fiberoptic bronchoscope could safely remove the cancerous tissues by point burning without destroying the adjacent normal tissues with high success rate. The ORR of the control group was 24.4%, and that of the experimental group was 63.6%, which was better than the control group. The AEs occurred during treatment in each group were of level 1 or level 2, and no serious life-threatening AEs occurred. Progression-free survival (PFS) time and overall survival (OS) time in the experimental group were both longer than those in the control group. Patients treated with MWA had a lower risk of disease progression and death than those treated with systemic administration alone., Conclusion: The treatment of lung cancer using systemic administration combined with MWA under CT and fiberoptic bronchoscope is more effective than using systemic administration alone, which can be promoted in clinical treatment., Competing Interests: Declaration of competing interest The authors declare that they have no potential conflicts of interest., (Copyright © 2021. Published by Elsevier Taiwan LLC.)

Albumin is the most abundant serum protein that transports hormones, free fatty acids, bilirubin, various ions, and drugs. The current study investigated the potential application of laser-induced fluorescence (LIF) in clinical analysis of human serum albumin (HSA) as a biomarker of liver and kidney disease. The excitation wavelength and HSA concentrations were systematically varied, and the LIF spectrum was recorded, in order to construct a standard calibration curve. Over a wide range of excitation wavelengths, excitation at 350 nm showedthe maximum fluorescence emission centered at 500 nm. We compared the determination of certain concentrations of HSA using both LIF and conventional laboratory assays. The LIF technique proved to be highly accurate and efficient. It may be concluded that femtosecond LIF provides a new, easy, very sensitive, precise, and direct method of detecting albumin in various biological samples., Competing Interests: Declaration of Competing Interest 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., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Cortisol and cortisone are common markers for stress and thus preferentially analyzed in matrices that allow non-invasive sampling such as saliva. Though the major drawback of immunoassays is lack of specificity due to cross reactivities, they are still most commonly used for quantification of steroid hormones. To overcome such problems, sensitive methods based on liquid chromatography-mass spectrometry are becoming more and more accepted as the golden standard for steroid bioanalysis as they achieve accurate quantification at trace levels for multiple analytes in the same run. Along this line, the aim of this study was the development of a new microflow UHPLC-ESI-MS/MS method for the measurement of salivary cortisol and cortisone, which due to its microflow regime provides enhanced sensitivity and is more ecofriendly. The developed method implemented sample preparation by Solid-Phase Extraction (SPE) in a 96-well plate format. Data acquisitions were carried out in MRM (multiple reaction monitoring) mode. The quantitative determination of endogenous compounds in saliva remains a challenge since analyte-free matrix is lacking. Hence, a surrogate calibrant approach with cortisol-d 4 andcortisone- 13 C 3 was applied for the target compounds in the presented method. A number of factors were optimized and the method validated. The lower limit of quantitation (LLOQ) was 72 and 62 pg mL -1 for cortisol and cortisone, respectively. Linear calibration was achieved in the range from 0.062 to 75.5 ng mL -1 for cortisol-d 4 and 0.072 to 44 ng mL -1 forcortisone- 13 C 3 . The performance of the method was also evaluated via proficiency test for salivary cortisol. Finally, it was applied successfully to evaluate cortisol and cortisone concentrations in multiple batches in routine clinical stress study samples (4056 total injections with 1983 study samples). Moreover, the instrument performance (in particular retention time variability) within each batch, between different batches and lot-to-lot of 5 investigated capillary columns over time is described. The work documents that micro-UHPLC-ESI-MS/MS is suitable and robust enough to carry out a full clinical study with greater than 1000s of samples over an extended period if adequate internal standards can be used., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Bone marrow necrosis (BMN) is a rare secondary disorder of many discrepant neoplastic processes. The etiology is diverse, and malignancy is the most common background disease., Patients and Methods: Between 2005 and 2019, a total of 23 cases of BMN were detected and analyzed at Zhujiang Hospital and Nanfang Hospital., Results: In our study, the 40-60-year-old age group was the one with the highest incidence of BMN (n = 12, 52.2%). The background diseases of patients with BMN varied. Eighteen (78.3%) of 23 patients were diagnosed with hematologic diseases at the same time, most of which were acute B lymphocytic leukemia (n = 8, 34.8%). The complete blood count of these 23 patients noted a decrease in hemoglobin (100%), a decrease or increase in white blood cells and neutrophils, and thrombocytopenia (78.3%). The levels of lactate dehydrogenase (> 300 U/L) and serum ferritin (> 500 μg/L) were elevated in all patients, and 16 (94.1%) of 17 patients presented with increased d-dimer levels. The 2-week cumulative survival and 2-year cumulative survival of patients with BMN were 56.5% and 47.4%, respectively. The mortality probability within 2 weeks was 43.5%, and the adjusted mortality probability was 26.7% within 2 weeks to 2 years, indicating that patients with BMN had the greatest risk of death within 2 weeks., Conclusion: BMN patients with B lymphocytic leukemia as the background disease had a better prognosis than those with other background diseases. BMN of unknown etiology may have an extremely poor prognosis. Therefore, diagnosing the background disease plays an important role in the treatment of BMN., (Copyright © 2020. Published by Elsevier Inc.)

This review article summarises aspects of the determination of amino acids using capillary and chip electrophoresis in combination with contactless conductivity detection from their historical beginnings to the present time. Discussion is included of the theory of conductivity detection in electromigration techniques, the design of contactless conductivity cells for detection in capillaries and on microchips, including the use of computer programs for simulation of the conductivity response and the process of the electrophoretic separation of amino acids. Emphasis is placed on optimisation of the background electrolyte composition, chiral separation, multidimensional separation, stacking techniques and the use of multidetection systems. There is also a description of clinical applications, the determination of amino acids in foodstuffs, waters, soils and composts with emphasis on modern techniques of sample treatment, such as microdialysis, liquid membrane extraction and many other techniques., (Copyright © 2020 Elsevier B.V. All rights reserved.)

During the last few decades, there has been evident growth in the development and fabrication of nano structured materials due to their wide application for different fields of material science and engineering. Among all these materials, natural polymer-based hydrogels such as polysaccharides, proteins, lipids etc. are gaining significant attention in the field of nanotechnology. Among them Chitosan hydrogels are widely used. These are cross-linked hydrophilic polymers which facilitate the uptake of significant quantity of water without being dissolved in water. They are hydrophilic in nature antimicrobial mainly need for fabrication of novel biomaterial These are chemically and physically firm, soft, stretchy polymeric network, reusable and multi-functional in nature. Chitosan has extensive properties including electrical, photo thermal characteristics, catalysis, anti-microbial characteristics, drug degradation, pollutants removal; bio-sensing etc. make it ultimate for "smart" nanostructure material. Recently, advance in the applications of Chitosan has made it an appreciable research material among the researchers. In this review we will focus on properties and applications of Chitosan as smart material in sensor configurations in various fields like clinical and environmental analysis., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Purpose: To develop a fast and accurate convolutional neural network based method for segmentation of thalamic nuclei., Methods: A cascaded multi-planar scheme with a modified residual U-Net architecture was used to segment thalamic nuclei on conventional and white-matter-nulled (WMn) magnetization prepared rapid gradient echo (MPRAGE) data. A single network was optimized to work with images from healthy controls and patients with multiple sclerosis (MS) and essential tremor (ET), acquired at both 3 T and 7 T field strengths. WMn-MPRAGE images were manually delineated by a trained neuroradiologist using the Morel histological atlas as a guide to generate reference ground truth labels. Dice similarity coefficient and volume similarity index (VSI) were used to evaluate performance. Clinical utility was demonstrated by applying this method to study the effect of MS on thalamic nuclei atrophy., Results: Segmentation of each thalamus into twelve nuclei was achieved in under a minute. For 7 T WMn-MPRAGE, the proposed method outperforms current state-of-the-art on patients with ET with statistically significant improvements in Dice for five nuclei (increase in the range of 0.05-0.18) and VSI for four nuclei (increase in the range of 0.05-0.19), while performing comparably for healthy and MS subjects. Dice and VSI achieved using 7 T WMn-MPRAGE data are comparable to those using 3 T WMn-MPRAGE data. For conventional MPRAGE, the proposed method shows a statistically significant Dice improvement in the range of 0.14-0.63 over FreeSurfer for all nuclei and disease types. Effect of noise on network performance shows robustness to images with SNR as low as half the baseline SNR. Atrophy of four thalamic nuclei and whole thalamus was observed for MS patients compared to healthy control subjects, after controlling for the effect of parallel imaging, intracranial volume, gender, and age (p < 0.004)., Conclusion: The proposed segmentation method is fast, accurate, performs well across disease types and field strengths, and shows great potential for improving our understanding of thalamic nuclei involvement in neurological diseases., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The quantitation of metanephrine (MN), normetanephrine (NMN), and 3-methoxytyramine (3-MT) - referred to as metanephrines -- by LC-MS/MS is the gold-standard for screening for pheochromocytoma and paragangliomas (PPGLs), tumours of the adrenal gland and the peripheral nervous system. An assay for metanephrines from dried blood spots (DBSs) would be of high clinical utility as it simplifies sample collection, enables remote sampling, and could increase compliance with the clinical recommendation for supine sampling. Moreover, DBS sampling facilitates the measurement of blood-derived metanephrines in pediatric patients - where DBSs are well-established - in order to diagnose neuroblastomas. Here, we adapted an established derivatization-based LC-MRM-MS assay for plasma catecholamines, and optimized the sample extraction, LC, and MS parameters to produce a fast, sensitive, and robust method for the measurement of metanephrines from DBSs, including 3-methoxytyramine. The DBS samples were excised, derivatized with phenyl isothiocyanate (PITC) on-spot, extracted, and measured by LC-MRM-MS. To validate assay suitability and performance, we assessed the linearity, precision, accuracy, recovery, and matrix effects of the method, and determined the stability of metanephrines in DBSs under different storage conditions. Assay performance for NMN, MN, and 3-MT was sufficient for quantitation from a single DBS within a linear range from 40 to 2000 pg/mL. MN and NMN were stable in DBSs for 2 weeks, whereas 3-MT was stable for one week regardless of storage temperature. Altogether, this work represents the first quantitative LC-MS/MS method for metanephrines from DBSs and provides a novel opportunity for the diagnosis of PPGLs and neuroblastomas in the future., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: CHB is the CSO of MRM Proteomics, Inc. and the CTO of Molecular You. The other authors declare no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Capillary electrophoretic separation of ketamine, norketamine, hydroxynorketamine, and dehydronorketamine was performed in the counter-current regime under the influence of oppositely-directed electroosmotic flow. For this purpose, the fused silica capillaries were covalently coated with the poly(acrylamide-co-3-acrylamidopropyl trimethylammonium chloride) copolymer (PAMAPTAC). The content of the cationic monomer APTAC in the polymerization mixture varied in the range 0-6 mol. % and the generated electroosmotic flow increased continuously in the 0-20 · 10 -9 m 2 V -1 s -1 interval. Importantly, it resulted in improved electrophoretic resolution of ketamine/norketamine, which increased from 0.8 for neutral PAM coating (i.e. 0% PAMAPTAC) to 3.0 for 6% PAMAPTAC. The determination of ketamine and its derivates in rat serum was performed in a 4% PAMAPTAC capillary with an inner diameter of 25 μm. The separation was performed in a 500 mM aqueous solution of acetic acid (pH 2.3). The clinical sample was deproteinized by the addition of acetonitrile to the serum and a large volume of the treated sample was injected directly into the capillary. The achieved limit of detection ranged from 2.2 ng/mL for dehydronorketamine to 4.1 ng/mL for hydroxynorketamine; the intra-day repeatability was 1.0-1.5% for the migration time and 2.8-3.3% for the peak area. The developed methodology was employed for time monitoring of ketamines in rat serum after intra venous administration of low doses of anaesthetic at a level of 2 μg per g of body weight., Competing Interests: Declaration of 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., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Drug-resistant bacteria is posing one of the greatest threats to human health. Carbapenemase-producing (CP) bacteria are a group of emerging highly drug-resistant bacteria which cause serious health problem in worldwide. Rapid and reliable detection of CP-bacteria is essential for point-of-care therapy and rapid infection control. In this study, a high-throughput tip-desorption electrospray ionization (tip-DESI) with solid-substrate tip was developed to couple ion mobility-tandem mass spectrometry (IM-MS/MS) for rapid screening of CP-bacteria from clinical samples. Raw bacteria spiked with indicators (i.e., carbapenems) was directly loaded on disposable substrate tip that connected with high voltage, and a desorption spray was applied for desorption and ionization of analytes. The substrate materials and desorption/ionization modes were optimized in this study. CP bacteria were ambiguously identified by monitoring of characteristic IM drift time and MS/MS spectra of hydrolyzed and decarboxylated carbapenems. We demonstrated this method for direct detection CP-bacteria in complex samples, showing excellent analytical performances including good tolerance to complex matrices, reducing interferences, high specificity, good repeatability, high sensitivity, and high analytical speed. Furthermore, this method was also applied for fast screening of CP-bacteria from different clinical bacteria, showing the potential applications for fast and reliable detection of antibiotic resistance in clinics., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A non-targeted lipidomics workflow based on C8 core-shell particle ultra high-performance liquid chromatography (UHPLC) hyphenated to ESI-QTOF-MS in data-independent acquisition (DIA) mode with sequential window acquisition of all theoretical fragment ion spectra (SWATH) was developed and applied to differential platelet lipidomics profiling of cardiovascular disease patients (stable angina pectoris (n = 10), ST-elevated myocardial infarction (n = 13)) against healthy controls (n = 10). DIA with SWATH generates comprehensive MS and MS/MS data throughout the entire chromatograms and all study samples. Hence, chromatograms can be extracted based on precursors or fragments which provided some benefits in terms of assay specificity in some cases. SWATH acquisition offers flexible experimental design with variable Q1 isolation windows. Liquid chromatography as well as SWATH settings were optimized to cover the lipidome of human platelets. The flexibility of the SWATH experiment design was utilized to implement target SWATH windows with narrow 5 Da Q1 precursor ion selection width (multiple reaction monitoring (MRM)-like SWATH windows) for the detection of low abundant oxidized phospholipids. Data processing was performed with MS-DIAL, and its feasibilities and caveats are discussed by illustrative examples. Thereby, identification of lipids is still a bottleneck in non-targeted lipidomics workflow. MS-DIAL, however, offers automatic identification via spectral matching using an in silico library. In total 1971 molecular features were detected cross the samples of which 611 were identified (total score >70%). The quality of the acquired data was validated with embedded quality control samples (n = 11). 80.3% of all features detected in the QC samples showed a coefficient of variation of below 30%. Multivariate statistics were used to visualize differences in the lipidome of distinct sample groups at a false discovery rate of 5%., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Poor drug solubility and bioavailability remain a significant and frequently encountered concern for pharmaceutical scientists. Nanoscaled lipid drug delivery systems (NSLDDS) have exhibited great potentials in oral delivery of poorly water-soluble drugs, primarily for lipophilic drugs, with several successful clinical products. In the past few years, we have find out that optimized composition of drug in lipid, surfactant, or mixture of lipid and surfactant omits the solubility, permeability and bioavailability issues, which are potential limitations for oral absorption of poorly water-soluble drugs. Lipids not only vary in structures and physiochemical properties, but also in their digestibility and absorption pathway; therefore selection of lipid excipients and dosage form has a pronounced effect on biopharmaceutical aspects of drug absorption and distribution both in vitro and in vivo. Therefore, in current critical review, a comprehensive overview of the different lipid based nanostructured drug delivery systems intended for oral administration has been presented. In addition, implication of in silico docking in designing of NSLDDS as well as mechanism of absorption of different lipid based nanoformulations through intestinal absorption window has also been offered. Moreover, attention has also been paid to NSLDDS that are currently undergoing preclinical or clinical analysis., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In spite of demonstrated lack of accuracy and consistency, quantification of steroid hormones is still most commonly executed via immunoassays. Mass spectrometric methods with triple quadrupole instruments are well established and, because of their proven robustness and sensitivity, best suited for targeted analysis. However, recent studies have shown that high-resolution mass spectrometers, like quadrupole time-of-flight instruments (QTOF), show comparable performance in terms of quantification and can generate additional sample information via untargeted profiling workflows. We demonstrate that adequate accuracy and selectivity for estradiol and testosterone can be achieved with a QTOF by data-independent acquisition with sequential window acquisition of all theoretical fragment-ion mass spectra (SWATH). Besides potential combination of targeted quantification and untargeted profiling, SWATH offers advantages with respect to sensitivity because the reduced total number of MS/MS experiments could be used to increase accumulation time without increasing cycle time. By applying a surrogate calibrant method leading to successful validation, a reliable method for absolute steroid quantification and high potential for steroid profiling has been developed. Linear calibration was achieved in the range from 10 to 1,000 pg mL -1 for 13 C 3 -estradiol and from 20 to 15,000 pg mL -1 for 13 C 3 -testosterone. Results for inter-day precision ( 13 C 3 -estradiol: 4.5-10.2%; 13 C 3 -testosterone: 5.1-7.8%) and inter-day accuracy ( 13 C 3 -estradiol: 94.6-112.8%; 13 C 3 -testosterone: 98.2-107.7%) were found to be well acceptable. Eventually, the method has been utilized to measure clinical samples of a study in which male volunteers obtained transdermal estradiol patches and sex hormone levels were quantified in plasma., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The first step to determine a medical or surgical intervention is the clinical exam. The clinical exam is the paramount step to provide a treatment plan that can be modified and individualized accounting the patient preferences. During the consultation for facial rejuvenation, attention should be paid to understand the patient's motivation and expectations. A thorough understanding of the anatomy and the natural history of ageing will facilitate the analysis of the face. Not only the degree of ptosis but also the degree of volume loss will need to be assessed, as well as the influence of the facial muscles, the skin quality, and the different causes of rythids. The comprehensive age-related anatomical changes are often perceived and described as tiredness or sadness. Patients very commonly only point out a single anatomical region. During the consultation, the surgeon should provide the patient with the information of the comprehensive interplay of the different anatomical regions and their individual ageing process. Obtaining a harmonious, natural appearing outcome is the result of excellent surgical skills and applied knowledge. The clinical exam should also find out traps and technical difficulties. Although standardized photographs allow a static evaluation of one's result, video may deliver additional information about the postoperative result, and may contribute to the understanding of the technique used. Spending the additional time by performing a thorough facial analysis and preoperative planning is well-invested time. Having a good understanding of the possible surgical improvements and limitations will be beneficial for both, the patient and the surgeon. Managing the expectations of the patient and careful preoperative planning will increase patient's satisfaction. At the same time, the surgeon will able to critically assess his/her own result and taking pleasure improving their own technique., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

A capillary electrophoresis approach with capacitively coupled contactless conductivity detection method has been developed for the determination of inorganic metabolites (thiocyanate, nitrite and nitrate) in human saliva. Field amplified sample injection, as a simple sample stacking technique, was used in conjunction for online preconcentration of above inorganic anions. A selective separation for the target anions from other coexisting constituents present in saliva could be obtained within 14min in a 10mmol/L His-90mmol/L HAc buffer (pH 3.70) at the separation voltage of -18kV. The limits of detection and limits of quantification of the three analytes were within the range of 3.1-4.9ng/mL (S/N=3) and 10-16ng/mL (S/N=10), respectively. The average recovery data were in the range of 81-108% at three different concentrations. This method provides a simple, rapid and direct approach for metabolite analyses of nitric oxide and cyanide based on noninvasive saliva sample, which presents a potential fast screening tool for clinical test., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objectives: To investigate the dynamics and clinical significance of hepatitis B surface antigen (HBsAg) levels in hepatitis B e antigen (HBeAg)-negative acute-on-chronic liver failure (ACLF) patients receiving lamivudine., Methods: Fifty-nine nucleoside-naïve patients with HBeAg-negative ACLF were enrolled and treated with 100mg of lamivudine daily. The dynamics of serum levels of HBsAg and HBV DNA were analyzed at baseline and at week 12, or before death if the patient died within 12 weeks., Results: Twenty-eight cases died within 12 weeks and 31 cases survived after treatment. HBsAg levels of patients who survived were significantly higher than those of patients who died both pre- and post-treatment (all p<0.05). There were 33 patients with pretreatment HBsAg levels above 4000 COI and 26 cases with levels below this value. The 12-week survival rate of those with levels ≥4000 COI was significantly higher than that of patients with levels <4000 COI (66.7% (22/33) vs. 34.5% (9/26), Chi-square = 5.991, p=0.014). Regardless of the pretreatment HBsAg level, there was no significant difference in HBV DNA levels between survivors and those who died, and no correlation was observed between HBV DNA and HBsAg (all P>0.05)., Conclusions: Changes in HBsAg level could be a useful parameter for predicting the short-term outcome of lamivudine monotherapy in HBeAg-negative ACLF., (Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.)