Multiple Roles of LOXL2 in the Progression of Hepatocellular Carcinoma and Its Potential for Therapeutic Targeting.

LOXL2, a copper-dependent amine oxidase, has emerged as a promising therapeutic target in hepatocellular carcinoma (HCC). Increased LOXL2 expression in HCC has been linked with an aggressive phenotype and represents a poor prognostic factor. Here, we focus on the mechanisms through which LOXL2 orchestrates multiple oncogenic functions in HCC development. We performed a review of the current knowledge on the roles LOXL2 performs in the modulation of the HCC tumor microenvironment, formation of premetastatic niches, and epithelial-mesenchymal transition. We also highlighted the complex interplay between LOXL2 and hypoxia, angiogenesis, and vasculogenic mimicry in HCC. At the end of the review, we summarize the current LOXL2 inhibitors and discuss their potential in HCC precision treatment.

Association of Peritoneal Cytology with Other Prognostic Factors in Endometrial Cancer.

Purpose: It is still debatable whether surgical staging of endometrial cancer (EC) should include sampling of peritoneal cytology (PC) and for what purpose this should be done. The aim of our study was to determine the significance of peritoneal cytology in EC and its association with other histological and clinical parameters. Methods: This is a retrospective study that comprises of results from 357 patients with EC that were operated in our center in the previous nine years. Patients were divided into two groups: the first group with a positive and the second group with a negative PC. Results: Malignant cells were found in the peritoneal cytology of 23 patients (6.4%), while 334 patients (93.6%) had negative PC. There was no significant difference in patients' age between the two groups (p = 0.20). Peritoneal cytology was more prevalent in the non-endometrioid than the endometrioid subtype of EC (p = 0.00). There was a significant statistical difference (p = 0.00) in malignant PC in stages where cancer is confined to the uterus (International Federation of Gynecologists and Obstetricians (FIGO) stages I and II) compared with those where cancer has metastasized outside the uterus (stages III and IV). Most of the patients with malignant PC (69.6%) had high-grade disease (G3). Conclusion: Malignant peritoneal cytology is associated with other negative prognostic factors in endometrial cancer (histological grade, FIGO stage, and non-endometrioid histological subtypes). Based on these findings, we encourage sampling of peritoneal washing in all EC patients and consider it mandatory in patients with non-endometrioid subtype, high-grade histology, and in advanced FIGO stage.

Bisphenol A and its analogues in outdoor and indoor air: Properties, sources and global levels.

Bisphenol A (BPA) and its analogues are high-volume production organic synthetic compounds used in the synthesis of plastics. BPA has been categorized as an endocrine disrupting compound due to its ability to disrupt the hormonal makeup of living organisms. Air and dust are common sources of exposure of BPA for living organisms and most sources are anthropogenic and a result of thermal destruction of BPA containing materials, import and export of recyclable materials (especially e-waste) and fugitive emissions near BPA handling facilities. Current reports on BPA levels in air are limited and focused on effluent and surface water analysis (due to BPA's propensity for environmental distribution to water). BPA's presence in the developing part of the world is of particular concern due to lack of regulations and uncontrolled incinerations of domestic and imported waste. The current review summarizes up-to-date scientific literature on BPA's occurrence in air, alongside physico-chemical and partitioning properties, persistence in air, seasonal variation, consideration of analytical strategies for BPA analysis and toxicological information. Globally reported air concentrations of BPA are included in this report, alongside reports on indoor air concentration of BPA and its analogues. As a special interest, levels of tetrabromobisphenol (TBBPA) are also mentioned. Overall, the highest outdoor air levels of BPA were reported in China (1.1 × 106 pg/m3) near a low-tech e-waste recycling site, while examination of indoor dust revealed the presence of bisphenol analogues used in "BPA-free" products, raising questions about their safety. Due to their low volatility, BPA and its analogues are mainly present in air associated with particles; this has important implications for their persistence in air and the role of particulate matter (especially microplastics) in their transport and deposition. Current understanding of BPA's particle association is limited, hence studying its potential for heterogeneous oxidative transformations is a pressing need required for accurate accounting of potential risk to human health and the environment.

Insights into sources and occurrence of oxy- and nitro-PAHs in the alberta oil sands region using a network of passive air samplers.

Mining-related activities in the Alberta Oil Sands Region (AOSR) are known to emit polycyclic aromatic hydrocarbons (PAHs) and related compounds to ambient air. This is a concern due to the toxicity of PAHs, including their transformation products such as nitrated (NPAHs) and oxygenated (OPAHs) PAHs. This is the first study that provided a more extensive outlook into the sources, occurrence in air, and spatial and seasonal patterns of NPAHs and OPAHs in the AOSR by using passive air sampling. A sampling campaign from 2013 to 2016 revealed concentrations of NPAHs that were much lower than those of OPAHs. The highest concentrations of NPAHs were concentrated in the region associated with extensive mining activities, with ∑NPAH concentrations ranging from 20 to 250 pg/m3. Within the oil sands (OS) mineable area, NPAHs associated with primary release appear more commonly, while NPAHs produced via oxidative transformation are predominant outside of this area. The concentrations of ∑OPAH ranged from 400 to 2400 pg/m3, with the highest air concentrations in the region located south of the main OS activity zone, with peak concentrations attributed to a 2016 forest fire event. Uptake of PAHs from ambient air and their subsequent conversion to generate OPAHs is believed to play an important role in wildfire emissions of OPAHs. The seasonal trend investigation was inconclusive, with NPAHs slightly higher during the winter, while OPAHs were slightly elevated during summer. A preliminary comparison of ambient concentrations of OPAHs and NPAHs in the AOSR to measurements in the Greater Toronto Area revealed a similar range of concentrations, but also a unique presence of certain NPAHs such as 4-nitrobiphenyl, 2-nitrodibenzothiophene, 2,8-dinitrodibenzothiophene and 6-nitrobenzo-(a)-pyrene. This indicates that AOSR might have its own NPAH profile - creating the need to better understand associated NPAH toxicity and propensity for long range transport.

Therapeutic drug monitoring of tranexamic acid in plasma and urine of renally impaired patients using solid phase microextraction.

The purpose of the research was to develop an improved solid phase microextraction (SPME)-based sampling protocol for the therapeutic drug monitoring of tranexamic acid (TXA) from plasma and urine of patients with chronic renal dysfunction (CRD) in order to correct the current dosing schedule to accommodate these patients. A 12-fold improvement in sampling efficiency (25 min for 96 samples -22 s per sample) was achieved with the use of hydrophilic-lipophilic balance (HLB)-coated SPME devices, thereby enabling high throughput profiling of TXA in the plasma and urine of 49 CRD patients undergoing cardiac surgery. A limit of quantification of 10 µg/mL and 25 µg/mL was obtained for plasma and urine respectively while a method accuracy of 103-105% and a precision of less than 8% was achieved. The results from this study were ultimately used by clinicians at the Toronto General Hospital to design a corrective pharmacokinetic dosing schedule for CRD patients. This green method further presents potential application in the clinical field for the fast high throughput monitoring of TXA not only in plasma but also in urine - a biological matrix seldom explored for the analysis of TXA - without the need for solvent-assisted extraction, extensive sample pre-treatment or clean-up, derivatization or excessive pH adjustment to improve amenability for analytical separation.

Untargeted metabolomics profiling of skeletal muscle samples from malignant hyperthermia susceptible patients.

PURPOSE: Malignant hyperthermia (MH) is a potentially fatal hypermetabolic condition triggered by certain anesthetics and caused by defective calcium homeostasis in skeletal muscle cells. Recent evidence has revealed impairment of various biochemical pathways in MH-susceptible patients in the absence of anesthetics. We hypothesized that clinical differences between MH-susceptible and control individuals are reflected in measurable differences in myoplasmic metabolites. METHODS: We performed metabolomic profiling of skeletal muscle samples from MH-negative (control) individuals and MH-susceptible patients undergoing muscle biopsy for diagnosis of MH susceptibility. Cellular metabolites were extracted from 33 fresh and 87 frozen human muscle samples using solid phase microextraction and Metabolon® untargeted biochemical profiling platforms, respectively. Ultra-performance liquid chromatography-high resolution mass spectrometry was used for metabolite identification and validation, followed by analysis of differences in metabolites between the MH-susceptible and MH-negative groups. RESULTS: Significant fold-change differences between the MH-susceptible and control groups in metabolites from various pathways were found (P value range: 0.009 to < 0.001). These included accumulation of long chain acylcarnitines, diacylglycerols, phosphoenolpyruvate, histidine pathway metabolites, lysophosphatidylcholine, oxidative stress markers, and phosphoinositols, as well as decreased levels of monoacylglycerols. The results from both analytical platforms were in agreement. CONCLUSION: This metabolomics study indicates a shift from utilization of carbohydrates towards lipids for energy production in MH-susceptible individuals. This shift may result in inefficiency of beta-oxidation, and increased muscle protein turnover, oxidative stress, and/or lysophosphatidylcholine levels.

RéSUMé: OBJECTIF : L'hyperthermie maligne (HM) est une condition hypermétabolique potentiellement mortelle déclenchée par certains agents anesthésiques et causée par une homéostasie calcique perturbée des cellules musculaires squelettiques. Des données probantes récentes ont mis en lumière une atteinte de diverses voies biochimiques chez les patients susceptibles à l'HM en l'absence d'anesthésiques. Nous avons émis l'hypothèse que les différences cliniques entre les individus susceptibles à l'HM et des témoins se refléteraient dans des différences mesurables de métabolites myoplasmiques. MéTHODE : Nous avons réalisé un profilage métabolomique d'échantillons de muscles squelettiques provenant de personnes négatives à l'HM (témoins) et de patients susceptibles à l'HM subissant une biopsie musculaire dans le but de poser un diagnostic de susceptibilité à l'HM. Les métabolites cellulaires ont été extraits de 33 échantillons de muscles humains frais et de 87 échantillons congelés à l'aide d'une microextraction en phase solide et des plateformes de profilage biochimique non ciblées Metabolon®, respectivement. La chromatographie en phase liquide à haute performance et la spectrométrie de masse à haute résolution ont été utilisées pour l'identification et la validation des métabolites, puis suivies d'une analyse des différences dans les métabolites entre les groupes susceptibles à l'HM et les groupes négatifs à l'HM. RéSULTATS : Des différences significatives ont été observées entre les groupes susceptibles à l'HM et les groupes témoins dans les métabolites issus de diverses voies (P : de 0,009 à < 0,001). Ces différences comprenaient l'accumulation d'acylcarnitines à longue chaîne, de diacylglycérols, de phosphoénolpyruvate, de métabolites de la voie d'histidine, de lysophosphatidylcholine, de marqueurs de stress oxydatif, et de phosphoinositols, aussi bien que des taux réduits de monoacylglycérols. Les résultats des deux plateformes analytiques concordaient. CONCLUSION : Cette étude métabolomique indique un changement de l'utilisation des glucides vers les lipides pour la production d'énergie chez les personnes susceptibles à l'HM. Ce changement pourrait entraîner une inefficacité de la bêta-oxydation, ainsi qu'une augmentation du renouvellement des protéines musculaires, du stress oxydatif, et/ou des taux de lysophosphatidylcholine.

Development of thin-film solid-phase microextraction coating and method for determination of artificial sweeteners in surface waters.

A semi-automated and sensitive method was developed for simultaneous determination of the six most consumed artificial sweeteners (AS) in surface waters using thin-film solid-phase microextraction (TF-SPME) and high-performance liquid chromatography (HPLC). A triple quadrupole mass spectrometer and an electrospray ionization source (ESI-MS) run in negative ionization and multiple reaction monitoring modes were employed for instrumental analysis. The TF-SPME method was optimized for the extraction phase, sample pH, desorption solvent, extraction time, and desorption time. In-house-synthetized-hydrophilic-lipophilic balance weak anion exchange (HLB-WAX) particles imbedded within a polyacrylonitrile (PAN) binder were selected as the extraction phase for the thin-film coating due to their cost-effectiveness and enhanced sensitivity for artificial sweeteners. Suitable analytical parameters that include linearity (R2 > 0.9914), recovery > 80%, inter, and intra-reproducibility less than 18% were obtained for the AS compounds studied. The developed method estimated limits of detection (LODs) ranging from 0.004 to 0.038 ng mL-1 The SPME method was successfully applied for the determination of ultra-trace levels of AS in water samples collected from Grand River (Ontario, Canada), downstream of three municipal wastewater treatment plants (WWTPs). Concentrations ranging from 0.03 to 20.3 ng mL-1 were found for the AS compounds studied.

High-throughput quantification of drugs of abuse in biofluids via 96-solid-phase microextraction-transmission mode and direct analysis in real time mass spectrometry.

RATIONALE: The workload of clinical laboratories has been steadily increasing over the last few years. High-throughput (HT) sample processing allows scientists to spend more time undertaking matters of critical thinking rather than laborious sample processing. Herein we introduce a HT 96-solid-phase microextraction (SPME) transmission mode (TM) system coupled to direct analysis in real time (DART) mass spectrometry (MS). METHODS: Model compounds (opioids) were extracted from urine and plasma samples using a 96-SPME-TM device. A standard voltage and pressure (SVP) DART source was used for all experiments. Examination of SPME-TM performance was done using high-resolution mass spectrometry (HRMS) in full scan mode (100-500 m/z), whereas quantitation of opioids was performed using triple quadrupole MS in multiple reaction monitoring mode and by using a matrix-matched internal standard correction method. RESULTS: Thirteen points (0.5 to 200 ng mL-1 ) were used to establish a calibration curve. Low limits of quantitation (LOQ) were obtained (0.5 to 25 ng mL-1 ) for matrices used. Acceptable accuracy (71.4-129.4%) and repeatability (1.1-24%) were obtained for validation levels tested (0.5, 30 and 90 ng mL-1 ). In less than 1.5 hours, 96 samples were extracted, desorbed and processed using the 96-SPME-TM system coupled to DART-MS. CONCLUSIONS: A rapid HT method for detection of opioids in urine and plasma samples was developed. This study demonstrated that ambient ionization mass spectrometry coupled to robust sample preparation methods such as SPME-TM can rapidly and efficiently screen/quantify target analytes in a HT context.

Solid phase microextraction coupled to mass spectrometry via a microfluidic open interface for rapid therapeutic drug monitoring.

Tranexamic acid (TXA) is an antifibrinolytic used during cardiac surgery that presents high inter-patient variability. High plasma concentrations have been associated with post-operative seizures. Due to the difficulties with maintaining acceptable concentrations of TXA during surgery, implementation of a point-of-care strategy for testing TXA plasma concentration would allow for close monitoring of its concentration during administration. This would facilitate timely corrections to the dosing schedule, and in effect tailor treatment for individual patient needs. In this work, a method for the rapid monitoring of TXA from plasma samples was subsequently carried out via biocompatible solid-phase microextraction (Bio-SPME) coupled directly to tandem mass spectrometry via a microfluidic open interface (MOI). MOI operates under the concept of a flow-isolated desorption volume and was designed with aims to directly hyphenate Bio-SPME to different detection and ionization systems. In addition, it allows the desorption of Bio-SPME fibers in small volumes while it concurrently continues feeding the ESI with a constant flow to minimize cross-talking and instabilities. The methodology was used to monitor six patients with varying degrees of renal dysfunction, at different time points during cardiac surgery. MOI proves to be a reliable and feasible tool for rapid therapeutic drug monitoring. Affording total times of analysis as low as 30 seconds per sample in its high throughput mode configuration while the single sample turn-around time was 15 minutes, including sample preparation. In addition, cross-validation against a standard thin film solid phase microextraction using liquid chromatography coupled to tandem mass spectrometry (TFME-LC-MS/MS) method was performed. Bland-Altman analysis was used to cross-validate the results obtained by the two methods. Data analysis demonstrated that 92% of the compared data pairs (n = 63) were distributed within the acceptable range. The data was also validated by the Passing Bablok regression, demonstrating good statistical agreement between these two methods. Finally, the currently presented method offers comparable results to the conventional liquid chromatography with acceptable RSDs, while only necessitating a fraction of the time. In this way, TXA concentration in plasma can be monitored in a close to real time throughput during surgery.

Miniaturized SPME tips directly coupled to mass spectrometry for targeted determination and untargeted profiling of small samples.

The analysis of small sample volumes, such as biofluids, tissues, and cells, has become increasingly popular over the past few years. In this paper, we introduce a sample-preparation tool that is suitable for such analysis, namely, a solid-phase microextraction (SPME) minitip featuring a tip apex (1 mm) that is coated with biocompatible polyacrylonitrile (PAN) and N-vinylpyrrolidone-co-divinylbenzene, also known as HLB particles. HLB particles are suitable for extracting a wide range of compounds with varying polarities, and the proposed SPME minitips were successfully used for the targeted (MS/MS) quantitation of polar, semi-polar, and nonpolar drugs of abuse (DoAs) in 1 µL of blood via offline nanoelectrospray ionization (nESI). Low limits of detection (LOD) and quantification (LOQ) were obtained for most of the targeted analytes (i.e. LODs between 0.5 and 2.5 ng mL-1). In addition, untargeted metabolomic profiling was performed on a single caviar egg (diameter ≤2.9 mm), with analysis being conducted via liquid chromatography high-resolution mass spectrometry (LC/HRMS). Clear chromatographic peaks were obtained despite the small size of the caviar eggs and the dilution introduced by LC. Multidimensional statistical analysis (PCA & PLS-DA) showed that the SPME minitips were able to successfully discriminate between samples, and the compounds that were tentatively identified corresponded with those expected to be found in fish. These findings highlight the importance of using suitable materials for pre-concentrating probe-sampling approaches, as they can provide analysts with sensitive instrumental response, particularly when studying the chemical profiles of small organisms.

Tranexamic Acid Dosing for Cardiac Surgical Patients With Chronic Renal Dysfunction: A New Dosing Regimen.

BACKGROUND: Tranexamic acid (TXA) is a common antifibrinolytic agent used to minimize bleeding in cardiac surgery. Up to 50% cardiac surgical patients have chronic renal dysfunction (CRD). Optimal dosing of TXA in CRD remains poorly investigated. This is important as TXA is renally eliminated with accumulation in CRD. High TXA doses are associated with postoperative seizures. This study measures plasma TXA concentrations in CRD cardiac surgical patients for pharmacokinetic modeling and dose adjustment recommendations. METHODS: This prospective cohort study enrolled 48 patients with stages 1-5 CRD, classified by Kidney Disease Outcome Quality Initiative. Patients were separated into 2 treatment groups. A "low-risk" group underwent simple aortocoronary bypass or single-valve repair/replacement and received a 50 mg/kg TXA bolus. A "high-risk" group underwent redo, aortic, multiple valve or combination surgery and received the Blood Conservation Using Anti-fibrinolytics Trial dosing regimen (loading dose 30 mg/kg, infusion 16 mg/kg/h with 2 mg/kg in pump prime). Primary outcome identified changes in TXA clearance and distribution volume, which provided the rationale for dose adjustment. Descriptive clinical outcomes assessed postoperative seizures, blood loss, ischemic-thrombotic complications, in-hospital mortality, and length of hospital stay. RESULTS: TXA concentrations were elevated and sustained above the therapeutic threshold for approximately 12 hours in high-risk stages 3-5 groups, in accordance to CRD severity. CONCLUSIONS: Using a pharmacokinetic model, we propose a simple new TXA dosing regimen that optimizes maximal antifibrinolysis and avoids excessive drug dosing.

Single-Use Poly(etheretherketone) Solid-Phase Microextraction-Transmission Mode Devices for Rapid Screening and Quantitation of Drugs of Abuse in Oral Fluid and Urine via Direct Analysis in Real-Time Tandem Mass Spectrometry.

The analysis of oral fluid (OF) and urine samples to detect drug consumption has garnered considerable attention as alternative biomatrices. Efficient implementation of microextraction and ambient ionization technologies for rapid detection of target compounds in such biomatrices creates a need for biocompatible devices which can be implemented for in vivo sampling and easily interfaced with mass spectrometry (MS) analyzers. This study introduces a novel solid-phase microextraction-transmission mode (SPME-TM) device made of poly(etheretherketone) (PEEK) mesh that can rapidly detect prohibited substances in biofluids via direct analysis in real-time tandem MS (DART-MS/MS). PEEK mesh was selected due to its biocompatibility, excellent resistance to various organic solvents, and its ability to withstand relatively high temperatures (≤350 °C). The meshes were coated with hydrophilic-lipophilic-balance particle-poly(acrylonitrile) (HLB-PAN) slurry. The robustness of the coated meshes was tested by performing rapid vortex agitation (≥3200 rpm) in LC/MS-grade solvents and by exposing them to the DART source jet stream at typical operational temperatures (∼250-350 °C). PEEK SPME-TM devices proved to be robust and were therefore used to perform ex vivo analysis of drugs of abuse spiked in urine and OF samples. Excellent results were obtained for all analytes under study; furthermore, the tests yielded satisfactory limits of quantitation (median, ∼0.5 ng mL-1), linearity (≥0.99), and accuracy (80-120%) over the evaluated range (0.5-200 ng mL-1). This research highlights plastic SPME-TM's potential usefulness as a method for rapidly screening for prohibited substances in on-site/in vivo scenarios, such as roadside or workplace drug testing, antidoping controls, and pain management programs.

Towards on-site analysis of complex matrices by solid-phase microextraction-transmission mode coupled to a portable mass spectrometer via direct analysis in real time.

On-site screening for target analytes in complex matrices, such as biofluids and food specimens, not only requires reliable and portable analytical instrumentation, but also solvent-free and easy-to-use sampling/sample preparation approaches that allow analytes of interest to be isolated from such matrices. The integration of sampling devices with field deployable instruments should be as efficient as possible, and should aim to provide rapid, precise, and accurate results that enable quick on-site decision. In this study, we evaluated solid-phase microextraction-transmission (SPME-TM) coupled to a portable single quadrupole MS system, via direct analysis in real time (DART), as an effective tool for the rapid screening of target analytes in biological and food matrices. Limits of quantitation (LOQ) in the low parts-per-billion levels (≤50 ng mL-1) were attained for most of the investigated analytes with total analysis times under 2 min per sample. Furthermore, we explored the suitability of this technology for on-site rapid molecular profiling of complex matrices. As a proof-of-concept, we demonstrate the rapid identification of milk samples from assorted animal and vegetal sources.