Form Of Platform Research Articles

Capture and detection of urine bacteria using a microchannel silicon nanowire microfluidic chip coupled with MALDI-TOF MS.

Fast bacterial identification in urine samples was achieved by capturing bacteria on a microchannel silicon nanowire microfluidic chip, followed by MALDI-TOF MS detection. Under the optimized conditions, bacteria with a concentration of 106 CFU mL-1 in urine samples could be identified without culture. If cultured for 4 hours, bacteria with a concentration as low as 103 CFU mL-1 were identified.

MALDI-TOF MS platform combined with machine learning to establish a model for rapid identification of methicillin-resistant Staphylococcus aureus

MALDI-TOF MS is an effective potential tool to distinguish between MSSA and MRSA. By combining the ClinProTools3.0 software and manual grouping intervention, we proposed a model optimization method for the first time. The cross validation of the model increased from 95.82% to 96.68%, and the accuracy of the model increased from 88.89% to 91.98%. Finally, we reported nine characteristic peaks of rapid detection of MRSA.

Proteomic Biomarkers to Guide Stratification for Covid‐19 Treatment: Exemplifying a Path Forward Toward Implementation?

Proteins and peptides serve as biomarkers in the context of multiple pathologies. The hypothesis that protein or peptide biomarkers may also be of value in the context of the Covid-19 pandemic appears self-evident. Proteome based biomarkers are not expected to display significant added value in the detection of viral infection but appear well suited to address a major unmet need: the prognosis of the course of disease, to guide appropriate, timely intervention. Based on similar approaches in the context of other diseases and using a CE-MS platform, urinary peptides are investigated for their value as biomarkers to assess disease progression after SARS-CoV-2 infection. The manuscript presented in this issue of Proteomics reports first results, indicating that urine peptides may be of substantial value in the assessment and prediction of severity of the Covid-19 disease course on an individual level. While the findings are not entirely surprising, the report does stand out from all others by a well-defined context-of-use, and, what is more, by presenting an already initiated validation study that may, if successful, result in immediate implementation of this proteomics-based diagnostic test. This approach should serve as positive example for the planning and execution of clinical proteomics studies.

Identification of selected primary bloodstream infection pathogens in patients attending Kisii level five and Homa Bay county hospitals

Background: Bloodstream infection (BSI) contributes to a substantial proportion of mortality in sub-Saharan Africa and is marked by the presence of bacterial and/or fungal microorganisms in the blood. Because BSI can be life threatening, it requires a timely, reliable and accurate diagnosis. This study retrospectively analyzed data of identified BSI pathogens and compared the performance of the different diagnostic technologies used in terms of accuracy, sensitivity, turnaround time (TAT) and cost. Methods: Currently, culture followed by analytical profile index biochemical strips (API), (BioMerieux) are used as the conventional standard diagnostics in Kenyan public hospitals and labs. We compared the results of this standard to that of the BioFire FilmArray (FA) (BioFire Diagnostics) and MicroScan WalkAway-40 plus System (MS) (Beckman Coulter) used in diagnosis of BSI. The FA technology was able to identify 150/152 bacterial and yeast isolates with an overall accuracy of 99.04% (95% CI: 96.59-99.88%), sensitivity of 98.68% (95% CI: 95.33-99.84%), mean TAT of 8 hours 40 minutes per eight samples and running cost per sample of USD 140.11. The MS identified 150/152 isolates with an overall accuracy of 98.56% (95% CI: 95.86-99.70%), sensitivity of 98.68% (95% CI: 95.30-99.84%), mean TAT per sample was 42 hours and running cost per sample of USD 28.05. API detected 150/152 isolates, with an overall accuracy of 99.04% (95% CI: 96.59-99.88%), sensitivity of 98.68% (95% CI: 95.33-99.84%) and the mean TAT per sample was 53 and 103 hours for bacterial and yeast samples, respectively, with a running cost per sample of USD 28.05.Conclusions: The findings in this paper suggest that the FA and MS platforms should be able to perform adequately in Kenya referral hospitals and medical clinics as a rapid diagnostic tool.

High resolution mass spectrometry provides novel insights into the ganglioside pattern of brain cavernous hemangioma

In this study we have optimized nanoelectrospray ionization (nanoESI) high resolution mass spectrometry (HR MS) performed on Orbitrap instrument in the negative ion mode for the determination of the composition and structure of gangliosides extracted from human brain cavernous hemangioma.The optimized HR MS platform, allowed the discrimination of 62 ions, corresponding to 52 different ganglioside species, which represents roughly twice the number of species existing in the current inventory of human brain hemangioma-associated gangliosides. The experiments revealed a ganglioside pattern dominated by GD-type of structures as well as an elevated incidence of species characterized by a low degree of sialylation and short glycan chains, including asialo GA1 (d18:1/18:0), which offer a new perspective upon the ganglioside composition in this benign tumor. Many of the structures are characteristic for this type of tumor only and are to be considered in further investigations for their potential use in early brain hemangioma diagnosis based on molecular markers. The detailed fragmentation analysis performed by collision-induced dissociation (CID) tandem MS provided information of structural elements related to the glycan core and ceramide moiety, which confirmed the molecular configuration of GD3 (d18:1/24:1) and GD3 (d18:1/24:2) species with potential biomarker role.

Olive Oil Quality and Authenticity Assessment Aspects Employing FIA-MRMS and LC-Orbitrap MS Metabolomic Approaches.

Edible vegetable oils comprise integral components of humans' daily diet during the lifetime. Therefore, they constitute a central part of dietary-exposome, which among other factors regulates human health. In particular, the regular consumption of olive oil (OO) has been largely accepted as a healthy dietary pattern. Responsible for its recognition as a superior edible oil is its exceptional aroma and flavor. Its unique composition is characterized by high levels of monounsaturated fatty acids and the presence of minor constituents with important biological properties, such as the so-called OO polyphenols. Being a high added value product, OO suffers from extensive fraud and adulteration phenomena. However, its great chemical complexity, variability, and the plethora of parameters affecting OO composition hamper significantly the selection of the absolute criteria defining quality and authenticity, and a reliable and robust methodology is still unavailable. In the current study, Flow Injection Analysis-Magnetic Resonance Mass Spectrometry (FIA-MRMS) was investigated under a metabolic profiling concept for the analysis of Greek Extra Virgin Olive Oils (EVOO). More than 200 monovarietal (Koroneiki) EVOO samples were collected from the main Greek OO producing regions and investigated. Both intact oil and the corresponding polyphenols were analyzed in fast analysis time of 2 and 8 min, respectively. In parallel, an LC-Orbitrap MS platform was used to verify the efficiency of the method as well as a tool to increase the identification confidence of the proposed markers. Based on the results, with FIA-MRMS, comparable and improved projection and prediction models were generated in comparison to those of the more established LC-MS methodology. With FIA-MRMS more statistically significant compounds and chemical classes were identified as quality and authenticity markers, associated with specific parameters, i.e. geographical region, cultivation practice, and production procedure. Furthermore, it was possible to monitor both lipophilic and hydrophilic compounds with a single analysis. To our knowledge, this approach is among the few studies in which two FT-MS platforms combining LC and FIA methods were integrated to provide solutions to quality control aspects of OO. Moreover, both lipophilic and hydrophilic components are analyzed together, providing a holistic quality control workflow for OO.

Narrow Aperture Detection Electrodes ICR Cell with Quadrupolar Ion Detection for FT-ICR MS at the Cyclotron Frequency.

Ion signal detection at the true (unperturbed) cyclotron frequency instead of the conventional reduced cyclotron frequency has remained a formidable challenge since the inception of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Recently, routine FT-ICR MS at the true cyclotron frequency has become a reality with the implementation of ICR cells with narrow aperture detection electrodes (NADEL). Here, we describe the development and implementation of the next generation of these cells, namely, a 2xNADEL ICR cell, which comprises four flat detect and four ∼45° cylindrical excite electrodes, enabling independent ion excitation and quadrupolar ion detection. The performance of the 2xNADEL ICR cell was evaluated on two commercial FT-ICR MS platforms, 10 T LTQ FT from Thermo Scientific and 9.4 T SolariX XR from Bruker Daltonics. The cells provided accurate mass measurements in the analyses of singly and multiply charged peptides (root-mean-square, RMS, mass error Δm/m of 90 ppb), proteins (Δm/m = 200 ppb), and petroleum fractions (Δm/m < 200 ppb). Due to the reduced influence of measured frequency on the space charge and external (trapping) electric fields, the 2xNADEL ICR cells exhibited stable performance in a wide range of trapping potentials (1-20 V). Similarly, in a 13 h rat brain MALDI imaging experiment, the RMS mass error did not exceed 600 ppb even for low signal-to-noise ratio analyte peaks. Notably, the same set of calibration constants was applicable to Fourier spectra in all pixels, reducing the need for recalibration at the individual pixel level. Overall, these results support further experimental development and fundamentals investigation of this promising technology.

Is pain temporary and glory forever? Detection of tramadol using dried blood spot in cycling competitions.

Tramadol is a synthetic opioid drug used in the treatment of chronic and acute pain. An abnormal prevalence of its misuse in elite sport to overcome pain resulting from prolonged physical effort was recently reported. However, besides its antinociceptive effects, tramadol consumption is associated with negative effects such as numbness, confusion, and reduced alertness. This fact prompted the Union Cycliste Internationale to ban the use of tramadol in cycling competitions. Herein, we present the development of a dried blood spot (DBS) sample collection and preparation method followed by a liquid-chromatography mass spectrometry (LC-MS) analysis to rapidly determine the presence of tramadol and its two main metabolites in blood samples. The detection window of each analyte was evaluated and the analysis of performance on various MS platforms (HRMS and MS/MS) was assessed. Tramadol and its two main metabolites were detected up to 12 h after the intake of a single dose of 50 mg of tramadol in positive controls. In professional cycling competitions, 711 DBS samples collected from 361 different riders were analysed using the developed methodology, but all returned negative results (absence of parent and both metabolite compounds). In the context of professional cycling, we illustrate a valid method bringing together the easiness of collection and minimal sample preparation required by DBS, yet affording the performance standards of MS determination. The proposed method to detect tramadol and its metabolites was successfully implemented in cycling races with a probable strong deterrent effect.

Development and validation of a custom panel including 183 Y‐SNPs for Chinese Y‐chromosomal haplogroups dissection using a MALDI‐TOF MS system

Y-chromosome SNP haplogroups exhibit geographic structuring in many populations around the world. Therefore, Y-chromosome haplogroups have been widely used to infer paternal biogeographical ancestry and high-resolution paternal lineage classification. In the present study, we designed a customized panel containing 183 Y-SNPs based on previous studies and evaluated the genotyping performance and repeatability, concordance, sensitivity, and ability of analysing case-type samples using a MALDI-TOF MS platform. The average call rate for duplicate typing of any one SNP in the panel was 97.0%. In the concordance and accuracy study, the results of haplogroup designation obtained from MALDI-TOF MS platform were fully consistent with those obtained from the next-generation sequencing (NGS) platform. The optimal amount of template DNA in the PCR seemed to be 10ng. However, if less DNA (≥156.25pg) was available, it was still possible to obtain meaningful haplogroup information. For the application part, this panel could be applied for the detection of blood, semen, and buccal swabs samples. Particularly, blood stain on FTA card samples could be dissected by direct PCR amplification on the MALDI-TOF MS platform. Besides, 371 unrelated male individuals from four Chinese ethnic groups (Han, Hui, Mongolian, and Kazak) were detected using this panel. Total 78 terminal haplogroups were found and the haplogroup diversity was 0.933576. The results demonstrate that this panel could be an accurate, fast, and cost-effective method for database construction where the amount of sample material is less of a concern and when the cost of the assay is taken into consideration.

Abstract 6482: Activity based protein profiling by SWATH/DIA-MS mode (ABPP-SWATH/DIA-MS) from OCT-embedded tissues biopsies reveals lung tumor-specific protease profiles

Abstract (a) Proteomics relies on protein digestion into smaller fragments/peptides and converge to proteome sequence information, classification, and expression levels without a screen of the cellular protein activities. Activity-based proteome profiling (ABPP) is emerging as a tool for functional proteomics based on mass spectrometry. We established an ABPP-SWATH/DIA MS platform compatible with OCT-embedded biopsies for differential global serine hydrolase (SHs) activity profiling and determine the Relative Depletion-Detected Activity (RDDA) index that reflects the relative activity of SHs in type IIIA lung adenocarcinoma biopsies. (b) To prevent digestion on streptavidin beads and MS spectra contamination, we used SWATH/DIA-MS2 signal intensities for the assessment of the ratio between the biopsy-extract solution either treated with DMSO or which had earlier been depleted with chemical probe for active enzyme molecules. We processed 24 lung biopsies (having a solution of depleted and non-depleted proteome) of long and short survivors with type IIIA lung adenocarcinoma tumor and their normal tissue counterparts and included peptides of 65 known endogenous inhibitors of SH. In parallel experiments, we analyzed the catalytically depleted part of the molecule by conventional ABPP-MS approach integrated on bead sample digestion. (c) We quantified 117 proteins annotated to the serine hydrolase family, of which as much as 60% was depleted with a CI of 90% in the biopsy-extract solution incubated with chemical probe. Among 133 enzymes monitored over the entire clinical cohort, we found as many as 89 enzymes with significant depletion that we further used to generate the activity SH profiles of each biopsy. We found 36 enzymes exclusively depleted in tumor tissue, both in short and long survivals, and with depletion absent from normal counterparts. By quantification of the catalytically active part of the molecule on the streptavidin bead sample pull-down digest, we found only 9 proteins that contained quality data for spectral counting comparisons. Partial Least Squares Discriminant Analysis (PLS-DA) analysis revealed the activity profiles of 9 enzymes perfectly discriminating the tumor tissues of short survivals, and these enzyme signatures were enhanced by the elevated activity of 2 enzymes - ADH1C and IAH1- in short-term survivors. (d) We have demonstrated that this approach can substantially increase the differential detection of protease activity between oncogenic lung tissue and their normal tissue counterparts. Citation Format: Tatjana Sajic, Stephan Arni, Rudolf Aebersold, Sven Hillinger. Activity based protein profiling by SWATH/DIA-MS mode (ABPP-SWATH/DIA-MS) from OCT-embedded tissues biopsies reveals lung tumor-specific protease profiles [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6482.

Ouabain regulates kidney metabolic profiling in rat offspring of intrauterine growth restriction induced by low-protein diet

AimsIntrauterine growth restriction (IUGR) can increase the risk of hypertension and kidney disease at adulthood due to fetal programming. In our previous study, we found that supplementation with low concentration of ouabain during pregnancy could restore glomerulus numbers at birth, rescuing kidney development. However, the metabolic pattern of kidney in IUGR offspring and the effect of ouabain have not been evaluated. Main methodsIn this study, based on GC–MS and LC-MS platforms, we used the protein restriction rat model to explore the molecular mechanisms of kidney damage induced by IUGR and the protective effect of ouabain. Key findingsThe results showed that malnutrition could induce IUGR in rat offspring at the 20th gestational day but ouabain treatment could partially reverse the body and kidney weight loss. Ouabain treatment could upregulate arginine, N-acetylornithine and carbamoyl phosphate as well as adenine nucleotide and guanine nucleotide downregulated by low-protein diet. Moreover, six metabolites were identified to be significantly correlated with fetal kidney weight, with 3 metabolites involved in arginine metabolism (arginine, N-acetylornithine, urea) and UDP-glucuronate correlated positively, while lysine and anthranilate correlated negatively. SignificanceThe results suggested that the underlying mechanism of ouabain against renal maldevelopment involved the metabolic regulation, particularly the arginine metabolism, which played an important role in the development of fetal kidney.

Identifying potential serum biomarkers of breast cancer through targeted free fatty acid profiles screening based on a GC-MS platform.

Recent advances suggest that abnormal fatty acid metabolism highly correlates with breast cancer, which provide clues to discover potential biomarkers of breast cancer. This study aims to identify serum free fatty acid (FFA) metabolic profiles and screen potential biomarkers for breast cancer diagnosis. Gas chromatography-mass spectrometry and our in-house fatty acid methyl ester standard substances library were combined to accurately identify FFA profiles in serum samples of breast cancer patients and breast adenosis patients (as controls). Potential biomarkers were screened by applying statistical analysis. A total of 18 FFAs were accurately identified in serum sample. Two groups of patients were correctly discriminated by the orthogonal partial least squares-discriminant analysis model based on FFA profiles. Seven FFA levels were significantly higher in serum from breast cancer patients than that in controls, and exhibited positive correlation with malignant degrees of disease. Furthermore, five candidates (palmitic acid, oleic acid, cis-8,11,14-eicosatrienoic acid, docosanoic acid and the ratio of oleic acid to stearic acid) were selected as potential serum biomarkers for differential diagnosis of breast cancer. Our study will help to reveal the metabolic signature of FFAs in breast cancer patients, and provides valuable information for facilitating clinical noninvasive diagnosis.

Healthy expectations of high hydrostatic pressure treatment in food processing industry

Fatty acids play critical roles in biological systems. Imbalances in fatty acids are related to a variety of diseases, which makes the measurement of fatty acids in biological samples important. Many analytical strategies have been developed to investigate fatty acids in various biological samples. Due to the structural diversity of fatty acids, many factors need to be considered when developing analytical methods including extraction methods, derivatization methods, column selections, and internal standard selections. This review focused on gas chromatography-mass spectrometry (GC–MS)-based methods. We reviewed several commonly used fatty acid extraction approaches, including liquid–liquid extraction and solid-phase microextraction. Moreover, both acid and base derivatization methods and other specially designed methods were comprehensively reviewed, and their strengths and limitations were discussed. Having good separation efficiency is essential to building an accurate and reliable GC–MS platform for fatty acid analysis. We reviewed the separation performance of different columns and discussed the application of multidimensional GC for improving separations. The selection of internal standards was also discussed. In the final section, we introduced several biomedical studies that measured fatty acid levels in different sample matrices and provided hints on the relationships between fatty acid imbalances and diseases. © 2019

Application of Derivatization in Fatty Acids and Fatty Acyls Detection: Mass Spectrometry‐Based Targeted Lipidomics

AbstractMass spectrometry (MS) has become an indispensable technique in the analysis of fatty acids (FAs) and fatty acyls due to its superior sensitivity and selectivity. Chemical derivatization is often introduced in MS‐based analysis because of the inherent poor ionization efficiency and selectivity of FAs and fatty acyls. This critical review presents an overview of commonly used MS platforms including gas chromatography‐MS, liquid chromatography‐MS, and mass spectrometry imaging combined with representative derivatization reagents for FAs and fatty acyls based on different chain‐lengths, saturation degrees, and conjugation groups. The method applications and limitations as well as critical derivatization conditions in different biological matrices are summarized. Special remarks on the chemical derivatization for MS‐based analysis are provided. Future perspectives associated with the detection of FAs and fatty acyls are discussed. This review provides valuable information and suggestions for the targeted detection of interested FAs and fatty acyls using different MS‐based platforms.

Effect of a lifestyle intervention program with energy-restricted Mediterranean diet and exercise on the serum polyamine metabolome in individuals at high cardiovascular disease risk: a randomized clinical trial

Many food items included in the Mediterranean diet (MedDiet) are rich in polyamines, small aliphatic amines with potential cardioprotective effects. The consumption of a MedDiet could increase polyamine concentrations. Based on experimental models, polyamine concentrations may be also influenced by physical activity (PA). We aimed to evaluate whether an intervention based on an energy-restricted MedDiet (er-MedDiet) and PA promotion, in comparison with an energy-unrestricted MedDiet and traditional health care, influences the serum pattern of polyamines and related metabolites in subjects at high risk of cardiovascular disease (CVD). This was a substudy from the PREDIMED-Plus trial, an ongoing randomized clinical trial including 6874 participants allocated either to an intensive weight-loss lifestyle intervention based on er-MedDiet, PA promotion, and behavioral support (er-MedDiet+PA group), or to an energy-unrestricted MedDiet and traditional health care group (MedDiet group). A total of 75 patients (n=38, er-MedDiet+PA group; n=37, MedDiet group) were included in this study. Serum concentrations of arginine, ornithine, polyamines, and acetyl polyamines at baseline and 26 wk of intervention were measured by an ultra-high-performance LC-tandem MS platform. At week 26, study groups had similar adherence to the MedDiet but patients randomly assigned to the er-MedDiet+PA group showed significantly lower mean energy intake (-340.3 kcal/d; 95% CI: -567.3, -113.4 kcal/d; P=0.004), higher mean PA (1290.6; 95% CI: 39.9, 2541.3 metabolic equivalent tasks · min/d; P=0.043), and higher mean decrease in BMI (in kg/m2) (-1.3; 95% CI: -1.8, -0.6; P<0.001) than the MedDiet group. However, no significant differences in serum polyamines or related metabolites were found between study groups after 26 wk of intervention and no significant between-group differences were found in glycated hemoglobin, HDL-cholesterol, or triglyceride concentrations. In individuals at high CVD risk, an er-MedDiet with increased PA did not result in significant changes of serum concentrations of polyamines or related metabolites in comparison with an energy-unrestricted MedDiet and no increase in PA. This trial was registered at isrctn.com as ISRCTN89898870.

Dual mass spectrometry as a tool to improve annotation and quantification in targeted plasma lipidomics.

High quality data, based on reliable quantification and clear identification of the reported lipid species, are required for the clinical translation of human plasma lipidomic studies. Lipid quantification can be efficiently performed on triple quadrupole (QqQ) mass spectrometers in targeted multiple reaction monitoring (MRM) mode. However, a series of issues can be encountered when aiming at unambiguous identification and accurate quantification, including (i) resolving peaks of polyunsaturated species, (ii) discriminating between plasmanyl-, plasmenyl- and odd chain species and (iii) resolving the isotopic overlap between co-eluting lipid species. As a practical tool to improve the quality of targeted lipidomics studies, we applied a Dual MS platform by simultaneously coupling a reversed-phase liquid chromatography separation to a QqQ and a quadrupole-time of flight (Q-ToF) mass spectrometers. In one single experiment, this platform allows to correctly identify, by high-resolution MS and MS/MS, the peaks that are quantified by MRM. As proof of concept, we applied the platform on glycerophosphocholines (GPCs) and sphingomyelins (SMs), which are highly abundant in human plasma and play crucial roles in various physiological functions. Our results demonstrated that Dual MS could provide a higher level of confidence in the identification and quantification of GPCs and SMs in human plasma. The same approach can also be applied to improve the study of other lipid classes and expanded for the identification of novel lipid molecular species. This methodology might have a great potential to achieve a better specificity in the quantification of lipids by targeted lipidomics in high-throughput studies.

Membrane Proteins — the Lipid Connection

We have used native MS of membrane proteins liberated form micelles and native membranes to study protein lipid interactions. Using different Orbitrap MS platforms developed for membrane proteins we show that it is possible to distinguish lipids, cofactors and drugs while in direct contact with membrane protein targets.We also showed how drug binding mediates phosphorylation in a class A GPCR (P2Y1R) and that purification of three further class A GPCRs results in binding of specific lipids that mediate downstream coupling.The ability to detect binding which has both structural and functional effects for example on lipid flippase activity or G‐protein coupling, is a powerful and important attribute of the study of membrane proteins by mass spectrometry.Very recently we have added yet further insights by ejecting membrane protein directly from their native membrane environments. Recent results will be presented.Support or Funding InformationWellcome Trust Investigator AwardERC Advanced Grant MRC Programme Grant

Serum metabonomics study on Cr (Ⅵ ) subchronic exposure rats based on UPLC-Q-TOF-MS/MS platform

Objective: To ananlyze the toxic effects and mechanisms of Cr (Ⅵ) subchronic exposure based on metabonomics techniques. Methods: Twenty-nine female Sprague-Dawley rats were randomly divided into control group, low dose group and high dose group, 10, 9, 10, respectively. The control group, low dose group and high dose group were treated with 0, 10, 50 mg/L Cr (Ⅵ) for 90 days respec tively. The serum samples of rats with different dose of Cr (Ⅵ) treatment were detected Using UPLC-Q-TOF-MS/MS technique and data was analyzed by PCA, PLS-DA and OPLS-DA to compare with metabolic profile in different Cr (Ⅵ) dose treatments. Pathway analysis was performed using MetaboAnalyst 4.0 software. Results: UPLC-Q- TOF-MS/MS has stable detection performance and reliable experimental data. The control group, low Cr (Ⅵ) and high Cr (Ⅵ ) metabolic profiles of rats serum differences was obviously, and there is significant difference of serum metabolic profile among rats treated with different dose of Cr (Ⅵ) . 18 differential metabolites were screened between Cr (Ⅵ) low dose group and control group, 23 differential metabolites between Cr (Ⅵ) high dose group and control group. Compared to control group, there were 13 differential metabolites in both Cr (Ⅵ) high dose group and Cr (Ⅵ ) low dose group, such as 3-Hydroxy-11Z-octadecenoylcarnitine, Anserine, Farnesyl pyrophosphate, Linoleoyl ethanolamid e, Linoleyl carnitine, Lithocholate 3-O-glucuronide, LysoPC [20∶2(11Z, 14Z) ], LysoPC[20∶3 (5Z, 8Z, 11Z) ], LysoPC[22∶2(13Z, 16Z) ], PG[16∶0/22∶5(7Z, 10Z, 13Z, 16Z, 19Z) ], PI[18∶1 (11Z) /20∶4(5Z, 8Z, 11Z, 14Z) ], PI[20∶3(5Z, 8Z, 11Z) /18∶0], Serotonin. These differential metabolites were related to Glycerophospholipid metabolism, Tryptophan metabolism, Pentose and glucuronate interconversions, Terpenoid backbone biosynthesis. Conclusion: Cr (Ⅵ) subchronic exposure could induce the significant difference of serum metabolic profile. The differential metabolites induced by Cr (Ⅵ) subchronic ex- posure were mainly related to amino acid and lipid metabolism.

Influence of Altitude on Phytochemical Composition of Hemp Inflorescence: A Metabolomic Approach.

The phytochemical profiling of hemp inflorescences of clonal plants growing in different conditions related to altitude was investigated. Four strains of industrial hemp (Cannabis sativa L., family Cannabaceae) of Kompolti variety were selected and cloned to provide genetically uniform material for analyses of secondary metabolites (terpenes, cannabinoids, and flavonoids) at two different elevations: mountain (Alagna Valsesia 1200 m ASL) and plains (Vercelli Province 130 m ASL). Environmental conditions influenced by elevation have proven to be important factors inducing variations in hemp inflorescences’ secondary metabolite composition. In fact, all plants grown at altitude exhibited a higher total amount of terpenes when compared with plains counterparts, with β-Myrcene, trans-Caryophyllene and α-Humulene as the main contributors. A metabolomic, un-targeted approach performed by HPLC-Q-Exactive-Orbitrap®-MS platform with subsequent data processing performed by Compound Discoverer™ software, was crucial for the appropriate recognition of many metabolites, clearly distinguishing mountain from plains specimens. Cannabidiolic acid CBDA was the most abundant phytocannabinoid, with significantly higher concentrations in the mountain samples. The metabolic pathway of CBGA (considered as the progenitor/precursor of all cannabinoids) was also activated towards the production of CBCA, which occurs in considerably 3 times higher quantities than in the clones grown at high altitude. Isoprenoid flavones (Cannaflavins A, B, and C) were correspondingly upregulated in mountain samples, while apigenin turned out to be more abundant in plains samples. The possibility to use hemp inflorescences in pharmaceutical/nutraceutical applications opens new challenges to understand how hemp crops respond in terms of secondary metabolite production in various environments. In this regard, our results with the applied analytical strategy may constitute an effective way of phytochemical profiling hemp inflorescences.

A conjunctive lipidomic approach reveals plasma ethanolamine plasmalogens and fatty acids as early diagnostic biomarkers for colorectal cancer patients

ABSTRACTBackground: Colorectal cancer (CRC) represents a third leading cause of cancer-related death worldwide. The reliable diagnostic biomarkers for detecting CRC at early stage is critical for decreasing the mortality.Method: A conjunctive lipidomic approach was employed to investigate the differences in plasma lipid profiles of CRC patients (n = 101) and healthy volunteers (n = 52). Based on UHPLC-Q-TOF MS and UHPLC-QQQ MS platforms, a total of 236 lipids were structurally detected. Multivariate data analysis was conducted for biomarkers discovery.Results: A total of 11 lipid species, including 1 Glycerophosphoethanolamine (PE), 3 ethanolamine plasmalogens (PlsEtn), 1 plasmanyl glycerophosphatidylethanolamine (PE-O), 3 fatty acids (FFA), 1 Fatty acid ester of hydroxyl fatty acid (FAHFA) and 2 Diacylglycerophosphates (PA) were identified to distinguish the CRC patients at early stage from healthy controls. In addition, these potential lipid biomarkers achieved an estimated AUC=0.981 in a validation set for univariate ROC analysis.Conclusion: By combining Q-TOF MS and QQQ MS analysis, the 11 lipids exhibited good performance in differentiating early-stage CRC and healthy control. This study also demonstrated that lipidomics is a powerful tool in discovering new potential biomarkers for cancer diagnosis.