= Emerging. More than 5 years before clinical availability.
= Expected to be clinically available in 1 to 4 years.
= Clinically available now.
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= Emerging. More than 5 years before clinical availability.
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= Expected to be clinically available in 1 to 4 years.
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= Clinically available now.
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Podium Presentations for |
To be presented in Track 3 (Papageno Hall) on Wednesday at 14:30
INTRODUCTION |
To be presented in Track 6 (Doppler Hall) on Thursday at 10:50 Mass spectrometry is a powerful tool to identify pathogens. However some issues such as mixture handling are usually beyond reach of whole-cell MALDI-TOF approaches. We developed a tandem mass spectrometry approach which addresses without a priori any kind of isolates but also more complex samples such as mixtures of organisms. Based on the analysis of the whole set of identified peptides from the sample, a phylogenetic information is quickly processed and presented in a format easily accessible for the clinician. An in-house developed microbiota reference standard comprising 24 different microorganisms was used for optimizing parameters for data acquisition by tandem mass spectrometry and data processing. Furthermore, the peptide signal is automatically search against a protein sequence database listing all the proteins involved in antibiotic resistances. This allows obtaining a quick pathogen identification and proposing an in silico antibiogram. This pipeline has been challenged with hundreds of clinical samples and resulted very robust. This work paves the way to wide proteotyping applications in the clinical lab. |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 14:50
Introduction. Podocyturia is a possible early sign of kidney abnormalities in patients. Currently, kidney damages are assessed using proteinuria measurements and the estimated glomerular filtration rate. Unfortunately, these parameters might not always be efficient in early detection of all patients. Most analytical techniques for the analysis of podocyturia are tedious, time-consuming, and may lead to results variability. We opted to develop a reliable methodology for podocyturia evaluation in patients with kidney involvement. |
To be presented in Track 3 (Papageno Hall) on Thursday at 14:30 This session will focus on three aspects of clinical analyses: 1) the discovery of biomarkers using semi- and untargeted metabolomic approaches; 2) the “de novo” structural elucidation of biomarkers; and 3) the quantitative analysis of biomarkers in the clinical field. The focus on biomarker discovery will be on the choice, collection and preparation of samples for optimal results and to prevent the risk to create artificial biases between the patient and control groups. Different aspects concerning the UPLC/MS analysis of samples will be addressed to increase the quality of the dataset and to obtain the necessary information to optimize the alignment (data mining) of the results. Other tools, such as various mathematical transformations applied to the marker peak areas, will be presented to improve biomarker discovery based on their abundance. Multivariate analyses, mainly the supervised orthogonal partial least-square discriminant analysis (OPLS-DA) statistical method, will show how to target biomarker candidates. In the second part of the seminar, tips on the structural elucidation of biomarkers by tandem mass spectrometry will be presented. Often, novel biomarkers are not listed in tandem mass spectrometry data banks. Therefore, tools will be presented to perform “de novo” structural elucidation of molecules. The issue related to the presence of structural isomers interfering with the interpretation of a biomarker fragmentation spectrum will be presented. The last part of the seminar will be dedicated to the development and validation of Multiple Reaction Monitoring (MRM) methods for the quantitative analysis of biomarkers. Two examples of UPLC methods optimization allowing the separation of the biomarker from structural isomer interferences will be presented as well as how to reduce matrix effects. During this Practical Training Track presentation, biomarker results for Parkinson’s disease and various genetic diseases such as Fabry and Gaucher diseases will be used as clinical applications. |
To be presented in Track 5 (Trakl Hall) on Wednesday at 11:00
Background: |
To be presented in Track 1 (Mozart 1-3) on Thursday at 11:10
Introduction: The adrenal cortex and gonads produce steroid hormones involved in salt and glucose homeostasis, blood pressure regulation, stress response and sex differentiation. These hormones are produced via a series of enzymatic steps and metabolites of steroids from each step are excreted and measurable in urine. Inborn disorders of steroidogenesis result from genetic mutations in distinct enzymes, causing a block in hormone production and lead to several forms of Congenital Adrenal Hyperplasia (CAH) and differences in sex development (DSD). Each enzyme deficiency is characterised by a distinct pattern of altered excretion of individual steroid metabolites relating to the specific enzymatic block. Ratios of urine steroid metabolites measured by gas chromatography-mass spectrometry (GC-MS) can be employed as surrogates of distinct steroidogenic enzyme activities. Widespread use of GC-MS multi-steroid profiling for rapid diagnosis of these disorders in the acute setting is often hampered by lack of specialist expertise. Here, we developed a novel steroid metabolomics approach for the detection and differentiation of inborn steroidogenic disorders, comparing its performance to that of conventional biochemical analysis by established steroid metabolite ratios. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 10:50
BACKGROUND: Several plasma proteins have been suggested |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 11:40
Background: We intended to develop a high-throughput LCMS-method to measure sub-picomolar levels of estradiol (E2) and estrone (E1). The method should be able to differentiate between pretreatment and suppressed blood levels of E2 and E1 in postmenopausal breast cancer patients undergoing aromatase inhibitor (AI) treatment, and should be well applicable in a routine lab. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 10:30
Introduction: In TDM and toxicology, the analyst is often confronted with complex problems, where results can have important clinical consequences. Untargeted screening is an analytical challenge, given the high number of molecules to be detected and the lack of standards available. Considered to be the reference method for screening, liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS) generates a large volume of high quality spectral data, with a lack of tools for visualizing and organizing MS data of these compounds. Here, we applied molecular networking for untargeted screening interpretation. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 10:30
INTRODUCTION: Reliable specific biomarkers are essential for the improvement of diagnosis and accelerated drug discovery. Biofluids like plasma and serum are unique sources of potential biomarkers, capable of revealing an organism’s status. However, despite major efforts and significant investment, only a limited number of new biomarkers stand up to the scrutiny of validation via the analysis of large sample cohorts. Serum extraction is a facile procedure capable of delivering a high consistency of processed samples independent of origin and operator. However, variables such as the collection tube material, blood clotting time, centrifugation speed and temperature etc., can have detrimental effects on the concentration of so-called "contaminating proteins" derived from blood cells. |
To be presented in Track 6 (Doppler Hall) on Thursday at 15:10
INTRODUCTION: The introduction of MALDI-ToF has reduced the time to identification of microbial isolates. However, it still requires a user to add a matrix to assist in ionisation. Furthermore, in some instances, such as for yeasts, additional extraction steps are required for accurate species level identification. REIMS has previously been demonstrated to provide accurate species-level classification of bacteria and yeasts direct from colonies; without the need for additional preparative steps. In comparison to MALDI-ToF based platforms, REIMS utilises the lipidomic profile of bacteria and fungi to determine their species-level classification without sample preparation which expands substantially the potential application of mass spectrometry to clinical microbiology laboratories. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 11:10
INTRODUCTION: Clofazimine (CFZ) is used as a second line drug for multidrug resistant tuberculosis (MDR-TB) treatment. It exerts slow bactericidal effect on mycobacteria by binding to DNA, leading to cell cycle disruption. CFZ is well absorbed orally with food but has a very long half-life. Its pharmacokinetics (PK) is not well understood especially in drug resistant TB patients where treatment monitoring & adherence play a vital role. There is limited literature on CFZ PK with only 3 - 4 labs testing CFZ drug levels globally. Efficient in vitro and in vivo activities against drug resistant strains and low rates of CFZ resistance have promoted its use in MDR-TB therapy. |
To be presented in Track 3 (Papageno Hall) on Wednesday at 9:20
Introduction |
To be presented in Track 6 (Doppler Hall) on Wednesday at 14:30
Background |
To be presented in Track 1 (Mozart 1-3) on Thursday at 14:50
INTRODUCTION: |
To be presented in Track 5 (Trakl Hall) on Wednesday at 11:40
Introduction |
To be presented in Track 5 (Trakl Hall) on Wednesday at 15:10
INTRODUCTION: Breast cancer (BC) is the most prevalent type of cancer in women in western countries. BC mortality has not declined despite early detection by screening, indicating the need for better informed treatment decisions. It is acknowledged that lipid metabolism is altered in tumours and cancer cells and that the heterogeneity of BC tumour subtypes is reflected in the expression levels of enzymes in lipid metabolism. Therefore, a novel non-invasive diagnostic tool for BC based on lipidomic analysis of plasma samples would allow for subtype-specific treatment and improved prospects for the patients. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 11:20
Background: Mass spectrometric steroid panels have potential for efficient single-sample methods for diagnosis of multiple disorders of steroidogenesis. Moreover within a specific disorder, such methods can facilitate subtyping for more rapid therapeutic intervention than currently allowed. Combined with advances in computational mathematics, such as machine learning (ML), it is now possible to move from traditional uni-dimensional approaches for interpreting diagnostic data to methods that can interpret patterns in multidimensional data. |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 14:30
INTRODUCTION: |
To be presented in Track 2 (Mozart 4-5) on Thursday at 9:10
Background |
To be presented in Track 1 (Mozart 1-3) on Thursday at 15:45
Introduction: Lactic acid is an organic acid found in two enantiomeric forms: the L- and D-lactate stereoisomers. D-lactic acid is produced from bacteria inhabiting the gut and also from mammalian methylglyoxal metabolism. In humans, the majority of systemic D-lactate is derived from the bacterial metabolism of carbohydrate in the upper GI tract. In pathological states, intestinal permeability is increased elevating the uptake of bacterial D-lactate from the gut. Circulating D-lactate concentrations may therefore provide a useful diagnostic tool for bacterial infections, gut permeability and GI health and disease. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 11:00
Background. LC-MS/MS is recommended for circulating steroid measurement, as its intrinsic specificity is supposed to guarantee accurate quantitation. LC-MS/MS also represents a promising opportunity to achieve harmonization of steroid measurements. However, LC-MS/MS assays applied in different laboratories use various pre-analytical and analytical approaches that may influence assay performance and final result. So far, little data have been generated on the comparability of LC-MS/MS steroid measurements. |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 11:00
INTRODUCTION: |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 11:00
INTRODUCTION: Uterine leiomyoma is one of the most frequent gynecological pathologies in women before the menopause. Despite numerous studies of factors involved in the genesis and growth of uterine leiomyomas the causes of uterine fibroids, as well as its recurrence are still the subject of discussion. It is of great interest to develop new noninvasive methods of early diagnosis of uterine leiomyomas and its recurrences using highly informative modern techniques such as mass spectrometry. Early prediction of leiomyoma recurrence will assist the patient and her gynecologist in deciding the most appropriate method of treatment. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 13:55
Introduction: Fetal sex has effects on prenatal stage, being male fetuses more prone to maternal stress than females. Fetus is protected from maternal cortisol by placental 11βHSD2 (by conversion to cortisone) and other enzymes that might inactivate cortisol into its metabolites. The effect of fetus gender on protection against maternal cortisol remains unexplored. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 13:15
Introduction: Risk assessment of new or existing chemicals finds a bottleneck in the evaluation of their potential as toxicants. Current approaches are too resource intensive in terms of time, money and animal use, thus limiting the number of substances which can be assayed. Chemical risk assessment using in vitro biological models such as human cell cultures allows to increase throughput while reducing cost and animal use. In such models, untargeted metabolomics can unveil triggered adverse outcome pathways (AOP) without the need for previous hypotheses. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 9:10
Introduction: |
To be presented in Track 3 (Papageno Hall) on Thursday at 10:30
Here I will be presenting three tracks that will hopefully help new users improve the quality and robustness of their assays. |
To be presented in Track 6 (Doppler Hall) on Wednesday at 9:20
INTRODUCTION: Early diagnosis is central in cancer care since it is directly related to the prognostic, evolution and survival of patients. For instance, the 5-year survival rate for breast cancer (BC) drops drastically if the pathology is diagnosed at a later state, from 80% at stage II to 22% at stage IV for metastatic cancer. |
To be presented in Track 6 (Doppler Hall) on Wednesday at 9:40
INTRODUCTION: |
To be presented in Track 3 (Papageno Hall) on Wednesday at 11:00
This is a practical training session describing the process of certifying traceable reference materials and/or traceable reference values of analytes in a clinical matricies, with a defined uncertainty. This work is routinely carried out at the NML to provide traceable values to materials listed on the JCTLM database, and also to provide traceable reference values to clinical materials. |
To be presented in Track 5 (Trakl Hall) on Wednesday at 9:00
Introduction |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 14:30
Background: Recent studies have shown that the results of immunoassay methods, using streptavidin-biotin interaction methodology, can be affected by the presence of high biotin circulating levels due to supplemental therapy leading to a growing number of adverse events. The evidences so far reporting on biotin interference by immunoassay methods are mainly based on case reports, rather than on experimental or clinical studies. |
To be presented in Track 5 (Trakl Hall) on Thursday at 14:30
Introduction: Today, most pathological tissue samples are stored as formalin-fixed paraffin embedded (FFPE) tissue blocks, which is the gold standard for histopathological analysis. This form preserves the tissue very well as formalin fixation forms methylene cross-bridges between proteins. Additionally, by paraffin embedding the tissue it allows for indefinite storage at room temperature so there are a number of large FFPE tissue archives around the world. Tissue microarrays (TMAs) is one of the most important forms of FFPE categories. TMAs are blocks that contain tissue cores from multiple sources in an array which allows for high-throughput analysis of many tissue samples. Mass spectrometry imaging (MSI) is a powerful tool that enables us to investigate the regional distribution of a variety of molecules in biological samples. It is crucial to have knowledge of the spatial distribution of biomolecules to understand the biological processes that take place within the tissues. Desorption electrospray ionization mass spectrometry imaging (DESI-MSI). particularly suited to investigate the spatial distribution of metabolites. Metabolite measurements are growing in importance as altered metabolism is a hallmark of many diseases, such as cancer. Currently, fresh frozen (FF) samples are preferred over FFPE samples in tissue based MSI studies. This is primarily due to the concern that small molecules will be lost during fixation and chemical processing as well as the belief that that paraffin will cause ion suppression. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 8:50
Introduction: |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 14:50
Introduction: |
To be presented in Track 5 (Trakl Hall) on Thursday at 13:35
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To be presented in Track 6 (Doppler Hall) on Thursday at 10:30
INTRODUCTION. Respiratory tract infections are a major cause of clinical treatment, but clinical assessments and standard clinical laboratory protocols are time consuming and often insufficient for reliable diagnoses. New methods, preferably applied directly to clinical samples, without any culturing, are needed to improve diagnostics, provide adequate treatment and reduce the use of antibiotics and associated development of antibiotic resistance. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 15:45
The fetus is thought to be protected from exposure to foreign antigens, yet CD45RO+ T cells reside in the fetal intestine. Here we combined functional assays with mass cytometry, single-cell RNA sequencing and high-throughput T cell antigen receptor (TCR) sequencing to characterize the CD4+ T cell compartment in the human fetal intestine. We identified 22 CD4+ T cell clusters, including naive-like, regulatory-like and memory-like subpopulations, which were confirmed and further characterized at the transcriptional level. Memory-like CD4+ T cells had high expression of Ki-67, indicative of cell division, and CD5, a surrogate marker of TCR avidity, and produced the cytokines IFN-? and IL-2. Pathway analysis revealed a differentiation trajectory associated with cellular activation and proinflammatory effector functions, and TCR repertoire analysis indicated clonal expansions, distinct repertoire characteristics and interconnections between subpopulations of memory-like CD4+ T cells. Imaging mass cytometry indicated that memory-like CD4+ T cells colocalized with antigen-presenting cells. Collectively, these results provide evidence for the generation of memory-like CD4+ T cells in the human fetal intestine that is consistent with exposure to foreign antigens. |
To be presented in Track 6 (Doppler Hall) on Thursday at 14:30
Introduction: Over the recent years, (multi-) drug resistant clinical isolates escalated resulting in a global threat that requires the fast and reliable detection of the corresponding resistance pattern. This is a major pre-requisite for an adequate and dedicated patient management and the prevention for further spread of those strains. We have developed a novel approach allowing MALDI-TOF mass spectrometry -based antibiotic susceptibility testing (AST), MBT FAST, within only a few hours. |
To be presented in Track 5 (Trakl Hall) on Thursday at 10:30 The majority of proteins that evolved after appearance of multicellular life are glycosylated and glycans significantly affect structure and function of these proteins. However, due to structural complexity of glycans and the absence of a direct genetic template, the analysis of protein glycosylation is much more complicated than the analysis of DNA or proteins. Consequently, the knowledge about the importance of individual variation in glycans for both normal physiological processes and diseases is still limited. In the last few years it is becoming increasingly clear that variations in a DNA sequence are only a beginning of the understanding of complex human diseases. Genetic polymorphisms have to be put in the context of complex biology of life and a more elaborate approach that combines different ‘omics phenotypes is needed to understand disease mechanisms and perform patient stratification that transcends genomics. Glycomics, as by far the most complex posttranslational modification, has an immense potential in this respect, which is only beginning to be investigated. By generating glycomic data for over 80,000 individuals from some of the best characterised clinical and epidemiological cohorts we enabled glycomics to meet other ‘omics. The analysis of this rich gold mind is painting a picture of a very complex genetic and epigenetic regulation of glycosylation that fine tunes protein activity in multiple biological systems and, if altered, contributes to development of different complex diseases. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 9:40
Background: Arterial hypertension is one of the most preventable causes of premature morbidity and mortality with resistant hypertension reported to be present in 5 to 30% of the total hypertensive population. Despite the poor prognosis, as many as 53% of those with resistant hypertension are reported to be non-adherent to their prescribed medication. This has led ourselves and others to develop LC-MS/MS methods to screen for the presence of antihypertensive drugs in urine. Despite the success of these assays, many suffer from not offering sufficient analytical breadth to cover all drugs prescribed in hypertension and/or not being able to identify key metabolites for some classes of drugs. Therefore we have developed a novel method for the detection of 49 commonly prescribed antihypertensive, antidiabetic and cardiovascular disease reducing medications in urine using LC-QTof MS. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 9:20 Irinotecan is a drug used as a chemotherapy agent mainly to treat colorectal cancer. Despite its remarkable efficacy, the gastrointestinal toxicity induced by the drug is life-threatening and often lead to an interruption of the treatment. Indeed, irinotecan is a pro-drug from which the active form is SN-38. Once reaching the liver, SN-38 is conjugated with glucuronic acid to then be excreted. However, once SN-38-G reaches the gut, a bacterial enzyme β-glucuronidase cleaves the glucuronic acid, releasing the aglycone SN-38 back into the GI tract, which is inducing the gastrointestinal toxicity. Even if inhibition of β-glucuronidase had proved to remove the ADRs from SN-38 in mice, it is logical to think that if this bacterial enzyme, which is present in more than 43% of the Human Microbiome Project Database species, might be affecting other metabolic pathways. In a project aiming to explore the potential to target the microbiome by selectively inhibit the bacterial β-glucuronidase, a 1H NMR spectroscopy and UPLC-MS metabolomics platform was used in order to assess the effect of this inhibitor on the metabolic profile of mice, particularly on the liver, gall bladder, colon tissues, plasma, urine, faeces and colon contents. A metabolic shift was observed only for the colon contents, faeces and urine samples, as well as a very slight one for the colon tissues. The metabolites significantly affected by the treatment seem to be mainly microbial metabolites, although metabolite identification process is still on-going. In another study where mice were treated with both the inhibitor and SN-38, the targeted UPLC-MS analysis of intestinal contents and plasma revealed the presence of different SN-38-G forms, potentially explained by a ring-opening mechanism leading to the carboxylate form of the molecule. Structural elucidation of these SN-38-G forms will be performed with high-resolution ion mobility mass spectrometry. Overall, even if further studies are needed to confirm preliminary findings, the inhibition of bacterial β-glucuronidase seems promising. |
To be presented in Track 5 (Trakl Hall) on Wednesday at 14:30
BACKGROUND & OBJECTIVES |
To be presented in Track 2 (Mozart 4-5) on Thursday at 13:15
The metabotropic glutamate receptor-1 (mGluR1) has recently been identified as an oncogene that is abundantly expressed in >60% of human melanoma, breast, renal cell and prostate cancer tissues. Many of these cancers cannot be successfully treated and new treatment modalities are clearly needed. In 2003, glutamate signalling through mGluR1 was reported to be a promising new molecular target for the treatment of cancer. We identified that affinity purified mGluR1 autoantibodies from plasma from a unique ataxia patient effectively block the mGluR1 receptor both in vitro and in vivo. These fully matured human antibodies are selective and have high target affinity. The precise sequence of these antibodies would offer an attractive basis for the development of a new antibody-based therapy for mGluR1 dependent cancers. |
To be presented in Track 5 (Trakl Hall) on Thursday at 9:10
INTRODUCTION: Over the past few years many studies have demonstrated the importance of altered glycosylation in tumor progression. These findings may contribute to discovery of biomarkers and treatment targets as well as understanding cancer biology. N-glycosylation of colorectal cancer (CRC) cell lines has been recently characterized revealing the association of antennary fucosylation with cell line differentiation and Caudal Type Homebox 1 (CDX1) expression. However, little is known about O-glycosylation of CRC cell lines due to its complexity, the presence of multiple isomeric structures as well as lack of enzymatic release methods making it overall challenging to perform in-depth analysis. OBJECTIVES: To provide a better understanding of the variation in O-glycosylation phenotypes and their association with other molecular features, an in-depth O-glycosylation analysis of 26 different colorectal cancer cell lines was performed. |
To be presented in Track 6 (Doppler Hall) on Wednesday at 11:00
The blood plasma proteome is maintained by influx and efflux of proteins from surrounding cells and organs. The liver secretes the majority of the highly abundant plasma proteins, the so-called classical plasma proteins, involved in the principal functions of plasma such as serving as transport medium, provide colloid osmotic pressure and maintaining hemostasis through the complement and coagulation systems. Blood plasma also contains numerous tissue proteins that most likely do not contribute to the principal functions of blood plasma. This group of proteins is more numerous than the classical plasma proteins and their role, if any, in the plasma is unclear. A subset of these proteins may be waste products resulting from the turnover of proteins and cells under physiological and pathological conditions. |
To be presented in Track 5 (Trakl Hall) on Wednesday at 14:50
Introduction: |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 11:20
Introduction |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 13:35
Background |
To be presented in Track 5 (Trakl Hall) on Wednesday at 11:20
Introduction |
To be presented in Track 5 (Trakl Hall) on Thursday at 13:15
Introduction: |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 14:30
The development of targeted proteomic assays, attempting to take biomarkers from the research lab to the clinic. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 08:30
INTRODUCTION |
To be presented in Track 3 (Papageno Hall) on Thursday at 08:30
Building multi-analyte LC-MS assays is a common temptation in Laboratories who are often faced with the drive to do “more with less”. This is especially true when the analytes in question are related clinically and share similar phys/chem properties. Adding these extra analytes can seem like additional data for free. However, in addition to extra logistical requirements (Increased cals, QCs, QA samples, data review, troubleshooting) there are analytical challenges to overcome such as differing stabilities and recoveries, different relative sensitivities, and the purity of reference standards that raise the question – how free is it really? |
To be presented in Track 6 (Doppler Hall) on Thursday at 13:35
Introduction: Volatolomics is the study of the volatile organic compounds (VOC) and one of the most fast-growing fields in non-invasive diagnostics. Several prospective clinical series have shown exhaled aldehydes, ketones, alkanes and short chain fatty acids to accurately identify patients with cancer. GC-MS primarily and online-MS, as proton transfer reaction-MS or selected ion flow tube-MS, are the most widely used platforms for VOC profiling. They have showed high coverage of the volatolome, low detection limits and high-throughput. However, they lack selectivity in several categories of chemical compounds (e.g. structural isomers). In these cases, multidimensional GC combined with high resolution MS can offer the required resolving power for the robust separation and determination. |
To be presented in Track 5 (Trakl Hall) on Thursday at 15:45 Skin depends on a unique profile of lipids that are necessary for the correct structure and function of the epidermal barrier, management of cellular communications and regulation of cutaneous homeostasis. Alterations in the cutaneous lipid profile can have severe consequences for skin health and such changes have been implicated in many inflammatory skin conditions. Using a targeted lipidomics platform we have investigated the prevalence of bioactive lipids in human skin, and reported an array of eicosanoids, octadecanoids, docosanoids, endocannabinoids, acyl ethanolamines and ceramides. We have conducted clinical studies and used human skin organ culture models and isolated cells in order to explore the differential contributions of lipid families to skin conditions. We have also explored lipid responses to various stimuli, and have examined temporal changes in lipid profiles aiming to understand their contribution to acute cutaneous inflammation and its resolution. Systemic supplementation with omega-3 polyunsaturated fatty acids (PUFA) has also revealed the differing cutaneous activities of these protective fatty acids, and demonstrated how they mediate their activities through perturbation of the profiles of existing species as well as formation of new lipids. Overall, we have shown that targeted lipidomics can elucidate the network of cutaneous bioactive lipids, support the development of biomarkers and diagnostics, and identify therapeutic targets for inflammatory skin disease. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 10:50
INTRODUCTION: Ruthenium metallocomplexes are promising candidates for anti-tumour therapy, which can be efficient even against tumours that are resistant to cisplatin. Cyclopentadienylbis(triphenylphosphine)ruthenium(II) chloride ([Ru(η5-C5H5)(PPh3)2Cl], RuCp in the text), as reported previously by us, presented in vitro significant cytotoxicity properties against several types of cancer cell lines. |
To be presented in Track 5 (Trakl Hall) on Thursday at 15:10
Introduction: Esogastric cancer is the fourth most commonly diagnosed cancer. Poorly cohesive carcinoma is sometimes difficult to identify via histopathology and presents a challenge for rapid and accurate diagnostics. The extent of the esogastric cancer cannot be deduced from the initial CT-scan or endoscopy exam, thus surgery needs to be performed followed by an intraoperative pathology exam. This exam is long and shows and error rate of about 30%. There is then an urgent need for a novel technology allowing for guided surgery and real-time diagnostic using specific molecular signatures in esogastric cancer while being of minimal invasiveness to the patient’s tissue. The water-assisted laser desorption/ionization mass spectrometry (SpiderMass) has demonstrated the capability to analyse biopsies ex vivo, allowing for correct classification of tumour type and grading (Saudemont et al. Cancer Cell 2018). Herein, we present a pipeline for ex vivo analysis using the SpiderMass system combined with Mass Spectrometry Imaging to asses and classify esogastric cancer. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 16:25
INTRODUCTION: Steroids play a crucial role in homeostasis of many biological processes including spermatogenesis, thus being responsible for some male infertility issues. Although steroids have been largely studied in many biological matrices (such as urine and plasma), there is very limited information of the steroid content in seminal liquid and its potential study as potential indicators of male infertility and other conditions. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 13:55
Non-Functionalized silica nanoparticles (SiNPs) are some of the widely used nanomaterial in diverse industrial sectors and nanoparticle based drug delivery applications. In industrial manufacturing environment SiNPs can possibly comes in contact with employees. Studies of cellular level toxicity effects of SiNPs in human cell lines are pivotal in the metabolite based biomarker discovery. Human lung epithelial cells (A549) are used to decipher the overall cellular level metabolic changes, non-targeted metabolite profiling with HILIC (hydrophilic interaction liquid chromatography) UPLC-HRMS method in a data dependent (DDA) mode was developed for this study. From the identified metabolome data and corresponding dysregulation in the metabolome of A549 biochemical pathways, our preliminary finding indicated 8 nm SiNPs elicit observable effects on the A549 cellular metabolism over larger SiNPs.The study identified some insights in early stage selective metabolite markers for nanomaterial related cytotoxicity in human cell line.Identified metabolites were annotated to pathways related to glutathione mediated detoxification, amino acid degradation, central carbohydrate metabolism and nucleotide metabolism with statistical significance (p < 0.01). |
To be presented in Track 1 (Mozart 1-3) on Thursday at 16:05
Faecal metabolomics allows for the non-invasive study of biomarkers in gastrointestinal (GI) disease. Current analytical techniques are limited in their applicability as they can lack in sensitivity (Nuclear Magnetic Resonance Spectroscopy) or require time-intensive sample preparation (Gas Chromatography - Mass Spectrometry). Here, we present the optimisation of LA-REIMS for faecal sample analysis and its implementation into a novel high-throughput application of LA-REI-MSI for the near-real time analysis and mapping of metabolites in whole fresh or frozen human faecal samples. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 08:30
Background / Objectives: Cervical cancer is the 4th most common cancer among women while its incidence is expected to rise by 43% in the UK by 2035 (Cancer Research UK, 2019). The microscopic examination of cervical cells currently carried out to screen asymptomatic women is prone to human error and can lead to high numbers of false-positives and false-negatives. Primary HPV DNA testing has been shown to be more accurate in screening and therefore is projected to replace cytology in the UK by the end of 2019 (Rebolj et al., 2019). Rapid evaporative ionization mass spectrometry (REIMS) is an innovative technique that allows interrogation of biological samples without any need for laborious sample preparation. Our main objective is to establish whether REIMS can be employed for the detection and grading of pre-invasive cervical changes. We also seek to assess the ability of REIMS to distinguish women with high-risk HPV (hrHPV) infection from women without infection. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 15:10
Introduction: Cancer is the second leading cause of death globally and is estimated to account for 9.6 million death in 2018 (WHO). Breast cancer (BC) remain the most common cancer in women and is ranked as the fifth amongst all cancers followed by colorectal, lung, cervix, and stomach cancers. The late diagnosis, using invasive and expensive procedures and the critical lack of medical and laboratorial infrastructures in the developing countries, are certainly two key contributing factors for this scenario. Therefore, more sensitive and specific diagnostic methods are urgently required. |
To be presented in Track 6 (Doppler Hall) on Wednesday at 15:10
INTRODUCTION: Matrix-assisted laser desorption and ionization mass spectrometry (MALDI MS) is suitable for clinical applications because of its performances. On the other hand, if organic matrices are used, analysis of small molecules (mass <1000 Da) becomes difficult due to a high number and intensity of matrix –related signals. Certain physiologically important molecules, such as hormones, carbohydrates, drugs, and others, belong to the small molecule group. Application of TiO2 nanoparticles as substrates for surface-assisted laser desorption and ionization (SALDI) MS is, in these cases, promising tool. |
To be presented in Track 6 (Doppler Hall) on Wednesday at 14:50
INTRODUCTION: Procalcitonin (PCT) is a 114 amino acids precursor of calcitonin produced by thyroidal C-cells and other neuroendocrine cells. PCT is clinically well recognized acute-phase protein biomarker, whose levels increase within 4 to 12 h upon stimulation. Compared to other sepsis markers, PCT could distinguish between infectious and non-infectious systematic inflammation or between viral and bacterial infections. Serum concentrations of PCT can vary from 0.1 ng/mL in healthy individuals to 120 ng/mL in patients with acute septic shock. In this study we show a technology for fast in-situ enrichment and mass spectrometry detection of PCT in human serum. The presented technology allows preparation of MALDI compatible protein chips by ambient ion landing. The electrosprayed proteins immobilized on conductive surfaces allow a wide range of bioanalytical assays. Compare to other materials, non-reactive surfaces suffer minimal nonspecific interactions with chemical species in the investigated sample and are thus an ideal substrate for selective protein chips. The reaction takes place directly on the protein chip and is followed by in-situ analysis by MALDI mass spectrometry. |
To be presented in Track 5 (Trakl Hall) on Thursday at 13:55
Introduction |
To be presented in Track 5 (Trakl Hall) on Thursday at 08:30
INTRODUCTION: Glycosylation is complex and highly abundant posttranslational modification of proteins that significantly affects their structure and activity. Glycans are directly involved in pathophysiology of every major disease and show great potential as clinical disease markers. |
To be presented in Track 6 (Doppler Hall) on Thursday at 14:50
Introduction: Differentiation between Vancomycin-Resistant and Vancomycin-Sensitive Enterococci (VRE and VSE) is of paramount important in the clinical microbiology laboratory for the correct management of infected patients. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 10:30
Background: Endothelial dysfunction (ED) contributes to diseases of the vasculature by influencing blood pressure, clotting and transport of fluids, nutrients and immune cells. Metabolic phenotypes associated with ED are not well characterized due to difficulties in assessing endothelial metabolism in situ. |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 11:40
INTRODUCTION: There is growing evidence that serum levels of apolipoproteins (apos) refines cardiovascular disease (CVD) risk assessment as compared to traditional blood-based lipid markers. Several research groups developed LC-MS strategies for quantitation of apos. An IFCC WG was recently established to set up a Reference Measurement System for 7 apolipoproteins. To this end the WG decided to develop a Reference Measurement Procedure using peptide-based primary Reference Materials and bottom up proteomics strategy. This strategy requires equimolar digestion of proteins to peptides to ensure metrological traceability, which is particularly challenging for apos C-I and C-III. The combined use of Lys-C and trypsin improves digestion for protein identification, but has not been evaluated for protein quantitation yet. Here we present the use of Lys-C in combination with trypsin to achieve equimolar digestion of apolipoproteins for bottom up proteomics quantification. |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 9:00
Introduction |
To be presented in Track 3 (Papageno Hall) on Thursday at 13:15
Key take-aways: |
To be presented in Track 2 (Mozart 4-5) on Thursday at 14:30
Introduction: |
To be presented in Track 6 (Doppler Hall) on Wednesday at 9:00
Background: Colon cancer is the fourth most common cancer in the UK resulting in approximately 16,000 deaths per year (Cancer Research UK, 2019). Biomarkers capable of risk stratification or leading to targeted therapy are therefore necessary to promote prompt patient diagnosis and treatment. This raises the need to understand cancer related metabolic pathways, and subsequently identify diagnostic and prognostic biomarkers. Overexpression of the gene TRIM44 has been related with carcinogenesis in a variety of cell lines and biospecimens, emphasising its importance as a potential cancer biomarker. However, TRIM44 mechanism of action is not yet elucidated. At the protein level, TRIM44 is structurally similar to TRIM29, which was previously shown to interact with p53, a crucial cell cycle regulator, by mediating its nuclear export. Taking into consideration the structural similarity between TRIM44 and TRIM29, this study hypothesised that TRIM44 mediates its carcinogenic effects is via p53-related pathway. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 15:10
Introduction |
To be presented in Track 5 (Trakl Hall) on Thursday at 14:50
INTRODUCTION: Mass Spectrometry Imaging (MSI) is an emerging technique in the field of histopathology which enables the capturing of molecular information of clinically important tissue samples, thus allowing the histological characterisation of tissues. Several techniques are currently available, for example Matrix Assisted Laser Desorption Ionisation, Secondary Ion Mass Spectrometry or Desorption Electrospray Ionisation, however all these techniques have some technical challenges (extensive sample preparation, sampling in vacuum or high voltage requirements). In this presentation a new technique, Laser Desorption Imaging – Rapid Evaporative Ionisation Mass Spectrometry (LDI-REIMS) is described as a new MSI technique suitable for the rapid analysis of a wide range of samples without any sample preparation and under ambient conditions. |
To be presented in Track 6 (Doppler Hall) on Thursday at 13:15
Title: |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 15:10
INTRODUCTION: A key metabolic alteration in tumour cells is increased dependency on glycolysis, resulting in the production of lactate which is transported out of cells by monocarboxylate transporters (MCT1 & MCT4) which are therefore a therapeutic target in cancer. Current literature suggests that inhibition of MCT1 in preclinical models can constrain cancer cell growth in tumours with low MCT4 expression. To date the systemic pharmacodynamic effects of the small-molecule non-competitive inhibitor of MCT1, AZD3965, the agent of study in this first-in-class (FIC) trial CRUKD12/004, have not been fully characterised. Preclinical metabolomics studies conducted at Imperial College London indicated that AZD3965 exposure caused increases in lactate, ketone bodies (also MCT1 substrates) and citrate in blood plasma and urine independently of tumour burden and tumour expression of MCT1, and also caused decreases in fatty acids in blood plasma. |
To be presented in Track 6 (Doppler Hall) on Thursday at 08:30
Introduction: Data review in a high-throughput clinical setting has traditionally involved the monotonous evaluation of every chromatogram at the expense of employees’ sanity. This review is as much art critique as data science, given the confidence to release a patient result more tangibly comes from placing eyes on picture of a chromatogram rather than a set of metrics. However, those eyes are looking for different things depending upon the reviewer, assay, day, week, month, and year, and may lead to inconsistency in data review and patient care. Further, the wealth of data from each patient that could provide insight into everything from instrument capacity to assay performance to personnel utilization remains largely unexplored. |
To be presented in Track 3 (Papageno Hall) on Wednesday at 9:40
INTRODUCTION: Mass spectrometry (MS)is an emerging technology that finds its way into medical laboratories for quantitation of proteins and proteoforms. Due to the complexity of bottom up proteomics quality requirements should be met during both method development and implementation as a lab-developed-test in order to guarantee that MS-based tests are fit-for-clinical-purpose(1). We previously developed a multiplexed LC-MRM-MS test for serum apolipoproteins (apo’s)(2). To maintain stable test performance, test quality has to be managed. Here we present long-term analytical performance data of an MS-based apo test in the real world setting of a medical lab. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 11:10
Introduction: For the past 60 years, the hematopoietic stem cell transplant (HSCT) has been successfully used as standard therapy for hematological disorders. After conditioning therapy, patients undergoing allogeneic HSCT present three phases of engraftment that occurs in different time points. The Pre-engraftment phase is defined by severe neutropenia, the next phase called engraftment occurs in the first 100 days after HSCT characterized by cellular immunodeficiencies and in the Post-engraftment phase the patients undergo an immune reconstitution of T cells, which normally occurs around 100 days or can take up to 2 years after allogeneic HSCT. Severe complications are associated with morbidity, mortality and malignances after allogeneic HSCT, which include effects in the oral cavity. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 8:50
INTRODUCTION: The human gut microbiota represents a pivotal environmental influence on the metabolism and overall health of the host. The immunomodulatory function of microbiota is mediated via the interaction of microbial metabolites with xenobiotic receptors expressed in immune cells and tissues. For instance, microbial catabolites of aromatic amino acids (i.e. tryptophan and tyrosine) were reported to activate pregnane X receptor (PXR) or aryl hydrocarbon receptor (AHR) in a ligand-specific fashion. The immune-metabolic homeostasis may be explored using quantitative metabolic profiling and targeted protein assays. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 14:30
The current regulatory rules governing in vitro diagnostic medical devices (IVDMD) in the EU are mainly represented by the European IVD directive dating back from 1998. This directive does not cover devices with characteristics related to newer techniques and applications in current in vitro diagnostic testing, lacks conformity with current international consensus with regard to the risk-based classification of IVDMD, lacks transparency with regard to the identification and labelling of IVDMD and lacks requirements for laboratory-developed tests (LDT). |
To be presented in Track 2 (Mozart 4-5) on Thursday at 14:50
Cervical cancer is preceded by cervical intraepithelial neoplasia of different degrees of severity (low grade squamous intraepithelial lesion, LSIL and high-grade squamous intraepithelial lesion, HSIL). None of the routine tests allow to assess the risk of neoplasia progression which is very important for young women. Cervicovaginal fluid (CVF) is a valuable source of clinical information about the physiological and pathophysiological status of the female reproductive tract. The aim of this study was to investigate the features of the CVF proteome in HPV-associated transformation of the cervical epithelium. |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 08:30 The C19 steroid 11β-hydroxyandrostenedione (11OHA4) is a major product of adrenal steroidogenesis, but was ignored for decades due to an apparent lack of activity. However, recent studies have demonstrated that 11OHA4 is the precursor to the potent 11-oxygenated androgens, 11-ketotestosterone and 11-ketodihydrotestosterone, that bind and activate the human androgen receptor with affinities and potencies similar to that of testosterone and 5α-dihydrotestosterone (DHT), respectively. The significance of these findings becomes apparent when considering androgen dependent diseases such as castration resistant prostate cancer and endocrine conditions associated with androgen excess such as polycystic ovary syndrome and congenital adrenal hyperplasia. Recent findings pertaining to the importance of the overlooked 11-oxygenated androgens will be presented, highlighting the role of 11-oxygenated androgens in disease states and challenging the paradigm that testosterone and DHT are the only clinically relevant androgens. |
To be presented in Track 2 (Mozart 4-5) on Thursday at 15:10
Introduction: The early-life development of the immune system and a specific low-grade inflammatory response may be monitored via quantification of inflammatory protein markers. Activated neutrophils are associated with calprotectins (CAL1 and 2) and myeloperoxidase (MPO), thus elevated abundance of those proteins in stool indicates the response against pathogens or auto-antigens. Similarly, eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) are released into the intestinal lumen from eosinophils activated by food allergens. In this study, we have developed multiplex protein assay for the absolute quantification of inflammatory proteins (ECP, EDN, CAL1, CAL2, MPO, and A1AT) and IgA in meconium/first stool samples from CELSPAC-TNG birth cohort study. |
To be presented in Track 3 (Papageno Hall) on Thursday at 15:45
INTRODUCTION: Method development for a quantitative LC-MS/MS method consists of several integrated steps involving many experimental factors which need to be simultaneously optimized to obtain maximum selectivity and sensitivity at minimum retention time. Optimization of experimental conditions for LC-MS/MS methods is usually performed by changing one-factor-at-time (OFAT) experiments. However, a much more effective optimization strategy for discovering important experimental factors and to optimize the responses is to utilize design of experiment (DoE). DoE is a systematic approach to data collection where all relevant experimental factors are studied simultaneously according to predefined plan. DoE can determine the individual and interactive effects of factors that can influence the output results of the measurements and estimate the optimum operating conditions for the LC-MS/MS method. |
To be presented in Track 4 (Paracelsus Hall) on Wednesday at 9:00
Introduction |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 9:20
INTRODUCTION: Cancer research is among the most studied areas of science and prostate cancer (PCa) is one of the most common types of cancer among men. Tissue samples, especially biopsies are often used in mass spectrometry based biomarker research as they have a great potential in understanding biochemical mechanisms underlying diseases such as cancer. Tissue microarrays (TMA) consist of several biopsies of different patients placed on a microscope slide. |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 11:20
INTRODUCTION:Acute Kidney Injury (AKI) is a frequent complication in hospitalized patients. Early detection of AKI is an unmet clinical need because current markers are delayed markers of functional kidney loss. Early kidney damage biomarkers such as neutrophil gelatinase-associated lipocalin (NGAL), tissue inhibitor of metalloproteinases 2 (TIMP-2) and insulin-like growth factor-binding protein 7 (IGFBP7) in urine are under investigation for early AKI detection and are typically measured by immunoassays. To increase specificity for kidney injury and to obtain information on the underlying pathology, kidney-specific proteins (e.g. uromodulin) and AKI biomarkers may be measured in a multiplex method. Here we aim to develop a multiplex LC-MRM-MS based method that is suitable for AKI biomarker quantitation. |
To be presented in Track 1 (Mozart 1-3) on Thursday at 10:50
INTRODUCTION: Processing and analysis of NMR metabolomics data typically involves a number of different software packages and that are partly controlled manually, limiting the reproducibility of the data processing workflow. |
To be presented in Track 3 (Papageno Hall) on Wednesday at 9:00
Background: So far, most publications reporting mass spectrometry-based measurement methods intended for diagnostic use describe in detail the method realization in one individual laboratory site - showing a very limited level of abstraction |
To be presented in Track 1 (Mozart 1-3) on Thursday at 13:35
Introduction |
To be presented in Track 1 (Mozart 1-3) on Thursday at 14:30
Introduction: There is a need to develop new diagnostic tools to diagnose and subtype patients with dysregulated steroidogenesis. Methods used today may require medication to be altered, multistep testing, imaging and even surgery to be able to make a final diagnosis. We hypothesize that it is possible to avoid many of these steps if the correct biomarkers or plasma steroid profiles are identified for patients suffering from these steroid dysregulation diseases, allowing diagnosis using only a single plasma sample. |
To be presented in Track 5 (Trakl Hall) on Thursday at 8:50
INTRODUCTION: Glycosylation is involved in many biological processes, one of these being fertilization. For example, literature indicates that alterations in N-glycan branching, sialylation and fucosylation are observed in seminal plasma of infertile males as compared to fertile males. While a causal relationship was not established, it is hypothesized that changes in glycosylation could disturb the normal course of fertilization. One of the major proteins in seminal plasma is the glycoprotein prostate-specific antigen (PSA). It acts as serine protease to liquefy seminal fluid by digesting seminal proteins, which results in an increased sperm motility. Moreover, it also plays an important role for the breakdown of cervical mucus and allows sperm to enter the uterus. A recent study screened seminal plasma proteins and revealed an up-regulation of fucosylation in subfertile men, which could be partially attributed to PSA. However, the exact role and effects of PSA glycosylation on male infertility is not yet clear. |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 11:40
INTRODUCTION: Burn injury can be a devastating trauma, affecting millions of people worldwide, with long-term personal and social implications for patients. Burns can result in life-limiting chronic pain, often refractory to treatment. Mechanisms behind burn injury are poorly understood and there has been little research into the molecular basis of burns and subsequent pain. It is important to explore the local signalling environment, but studies are often destructive in nature and preclude the collection of longitudinal temporal data. Microdialysis is a sampling method allowing the in vivo collection of solutes primarily from the extracellular interstitium and is ideal for the study of burn injury. Metabolomics offers an unbiased approach to the elucidation of metabolites involved in pathological events. Coupled with mass spectrometry, it provides a sensitive platform for the detection of metabolic changes due to burn injury. |
To be presented in Track 6 (Doppler Hall) on Thursday at 8:50
Introduction |
To be presented in Track 6 (Doppler Hall) on Thursday at 13:55
Introduction: There is a pressing need to develop new non-invasive screening tests for Oesophago-gastric (OG) cancer due to its high prevalence and poor survival. Previous studies have reported that urinary volatile organic compounds (VOCs) reflect human pathophysiological status. GC-MS based methods are the main approaches for urinary VOC profiling. However, biomarker discovery is limited by often inefficient and labour-intensive solvent extractions, by chromatographic resolution and by the unavailability of a complete, detailed, and high-throughput data-preprocessing methodology for large-scale untargeted VOC analysis of urine samples. Novel HiSorb sorptive extraction and conventional solid phase microextraction (SPME) are both tested and evaluated. GCxGC combined with TOF-MS is employed and offers outstanding identification capabilities. By coupling HiSorb/SPME with GCxGC-TOF-MS, this project aims to discover new predictive biomarkers for OG cancer. |
To be presented in Track 1 (Mozart 1-3) on Wednesday at 9:00
INTRODUCTION: Metabolite profiling – or metabolomics – presents a powerful global approach to measure shifts in metabolites as functional readouts of cellular state. Metabolites can complement upstream biochemical information obtained from genes, transcripts, and proteins and advance our understanding of how cells are altered in health and disease. Unfortunately, the great success in the characterization of genes, transcripts and proteins has currently no parallel in metabolites. Metabolomic studies are revealing large numbers of naturally occurring metabolites that cannot be characterized because their chemical structures and spectrometric data are not available. This is preventing metabolomics from evolving as fast as other omic sciences, and thus it is restricting the integration of multiple layers of omic data to gain more insights into the emergence of observed phenotypes. |
To be presented in Track 6 (Doppler Hall) on Thursday at 11:10
Rapid and accurate identification of microorganisms and their biomass |
To be presented in Track 2 (Mozart 4-5) on Wednesday at 9:40
Introduction: |
To be presented in Track 4 (Paracelsus Hall) on Thursday at 13:15
Introduction |
To be presented in Track 5 (Trakl Hall) on Wednesday at 9:40
Introduction: Fatty acids (FA) - organic compounds belonging to the group of carboxylic acids, the molecules of which contain a different number of carbon atoms (from 10 to 20). FA are divided into saturated (only single bonds), monounsaturated (with one double bond), and polyunsaturated (with several double bonds). The Omega-3 Index reflects the relative amount of omega-3 FA within red blood cell membranes. Many studies show that a low Omega-3 Index is associated with increased risk of cardiovascular disease (CVD), and it has been proposed that raising the index may help to reduce CVD risk. Measurements of FA in red blood cell membranes can provide important information about FA intake. In different regions of the world the value of the Omega-3 Index in the population varies considerably due to the peculiarities of nutrition. The purpose of this study was to identify the effect of age and gender on the Omega-3 Index in the average Caucasian population. |
To be presented in Track 5 (Trakl Hall) on Wednesday at 9:20
Background: There are some reference methods to determine serum total glycerol and triglyceride all by isotope dilution gas chromatography mass spectrometry which are time consuming and complicated. A need exists for a simple reference method that can be easily adopted to verify the accuracy of serum triglyceride measurements, especially with different measurement principles. Just as serum triglyceride concentrations are generally determined from total glycerol with or without a subtraction of free glycerol. So candidate reference methods involving isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) for total glycerol and free glycerol were established. The triglyceride concentration was the difference between total glycerol and free glycerol. |
Poster Presentations for |
Poster #4c View Map
INTRODUCTION: Colorectal cancer (CRC) is the third and second most common cancer occurring in men and women respectively. During CRC development, the tumor microenvironment has shown to be a key component to sustain progress and spreading of the tumor. The tumor microenvironment is a complex and dynamic matrix in which different cell types interact with each other to sustain tumor growth. In addition, this dynamic system shows fluctuation in physicochemical conditions. Hypoxia is a well-known state during tumor development responsible for metabolic adaptation, and promotion of tumor invasion. However, the role and mechanisms by which hypoxia induces lipid re-modeling are still not understood. Nevertheless, this regulation is related to tumor and stroma phenotype key players to better understand the tumor-microenvironment. |
Poster #13b View Map
BACKGROUND |
Poster #4b View Map
Introduction: Metanephrine and normetanephrine are the metabolites of adrenaline and noradrenaline, respectively. The determination of elevated levels of these compounds in plasma is used to diagnose the presence of pheochromocytoma and paraganglioma by a number of different methods, but mainly based on liquid chromatography coupled to mass spectrometry (LC-MS). A need for standardisation and harmonisation of plasma metanephrines (METs) analysis has been identified by the international pheochromocytoma and paraganglioma research support organization (PRESSOR). This is particularly challenging due to the low uncertainty and traceability required at the very low concentrations these compounds are typically measured at in plasma; normetanephrine 23-192 pg/mL and metanephrine 6-88 pg/mL(1). |
Poster #10d View Map
Introduction: For toxicological screening, methods not only need to be highly specific and sensitive but capable of identifying and quantifying a broad range of analytes in complex matrices. In this work a generic data independent acquisition (DIA) method was developed to detect and quantify a panel of drugs of abuse in human plasma using a high resolution accurate mass LC-MS/MS QTOF system. The method was designed as a generic data acquisition approach to extend the capability of LC-MS/MS analysis in routine clinical pathology for both targeted and untargeted data analysis. |
Poster #10b View Map
Introduction |
Poster #25b View Map
INTRODUCTION: Cortisol measurement is a key-test in the diagnosis of Cushing syndrome and in other clinically relevant conditions. It can be performed in different biological samples: serum, urine and, in more recent years, saliva. In salivary measurements only free hormone is detected, samples can be collected during normal daily routine and stress-induced cortisol release is less likely to occur than after venipuncture. Although immunometric assays for salivary cortisol are employed, but suffer from non-specificity due to cross reactivity with steroid metabolites, synthetic corticosteroids or drugs and lack of standardization between labs. |
Poster #14e View Map
Introduction: Vitamin D is hydrolized to its prohormone 25-hydroxyvitamin D in the liver, and being a predominant metabolite, circulating 25-hydroxyvitamin D serves as the preferred analyte that is monitored to determine nutritional status of Vitamin D. |
Poster #27e View Map
Introduction |
Poster #12b View Map
Introduction. Measurement of volatile organic compounds (VOCs) in breath is increasing in importance as a non-invasive diagnostic tool, especially for complex pathologies such as cancer [1]. Many methodological and clinical studies have been carried out to develop sophisticated new mass spectrometry methods and pinpoint differences in VOC levels between patients and controls. However, the origin of VOCs and their imbalance in certain diseases remain poorly understood. In a recent study from our group, cancer tissue headspace (HS) has been analysed ex-vivo to evaluate the in-loco production of VOCs [2]. In this study, we developed two different techniques of sample collection and analysis for VOCs in the HS of cells and organoids to create a new tool that will help clarify the role and production of VOCs in-vitro. |
Poster #25e View Map
Background: |
Poster #14h View Map
Background: Assessment of vitamin status is of clinical as well as nutritional importance. We wanted to develop a robust high-throughput LCMS-method for routine analysis and quantification of serum all-trans-retinol (Vitamin A) and α-tocopherol (Vitamin E). The method was based on comprehensive optimization of an assay currently in use at the Hormone Laboratory. |
Poster #24d View Map
Introduction: |
Poster #1c View Map
Introduction |
Poster #9a View Map
Introduction: Vitamin B12 deficiency can lead to potentially irreversible neurological symptoms such as memory deficits and gait ataxia. Up to 40% of older adults show metabolic abnormalities due to vitamin B12 deficiency. However, most adults have normal levels of serum B12 so that this condition often remains under-recognized. Metabolic B12 deficiency causes an elevation of serum methylmalonic acid (MMA), an expensive test rarely available in clinical settings. Urine MMA also increases in cases of B12 deficiency, but it is unclear whether it correlates with serum MMA and thus be reliable for diagnostic purposes. Also, various inborn errors of metabolism lead to increased MMA levels in urine and plasma. |
Poster #20a View Map
INTRODUCTION: Acylcarnitine profiling is routinely performed in order to quantitatively screen for disorders related to β-oxidation and / or organic acid metabolism. Today many laboratories still perform acylcarnitine profiling using LC-MS/MS based methodology that has remained almost unchanged for around 2 decades. This “classical” approach normally involves a direct infusion (no chromatographic separation) into the MS system following a butylation based sample derivatisation. Although this approach is still widely used today, improvement in LC-MS technology observed over the last 15 years such as UHPLC and higher sensitivity MS systems means that a non-derivatisation approach that can be more informative with a relatively rapid LC separation is feasible. |
Poster #17d View Map
Background: The male factor contributes to more than 50% of couples with conceive failure. Seminal plasma seems to be a promising noninvasive informative source for male infertility biomarkers. The possibility combined omics analysis of seminal plasma composition in defining the residual spermatogenesis during azoospermia was tested by the supervised OPLS-DA. Quantitative parameters of the model obtained turned out to be satisfactory to differentiate patients with residual spermatogenesis. |
Poster #24b View Map
Introduction: |
Poster #21a View Map
INTRODUCTION: Coated Blade Spray (CBS) is a technology that combines sample preparation and direct coupling to mass spectrometry (MS) on a single device. CBS is a coated stainless steel sheet with the shape of a small sword with an ultra-thin SPME coating that permits rapid enrichment of small molecules present in complex samples via free-concentration. As a substrate spray technology, it can generate instrumental signal on a mass spectrometer by supplying a small amount of organic solvent to the coated area of the device and then applying a strong electrical field to the non-coated area of the device, which in turn generates gaseous ions from a the tip of the CBS via electrospray ionization (ESI). |
Poster #1d View Map
INTRODUCTION: Genome wide association studies have identified the FTO gene as the first susceptibility gene of obesity. Previous studies have suggested a role of FTO in nucleic acid repair or modification, but how this leads to an alteration in energy homeostasis is unclear. In the present study, we utilized targeted metabolomics in an attempt to further elucidate mechanisms underlying the action of the FTO gene. |
Poster #13a View Map
Rationale: |
Poster #11g View Map
Introduction |
Poster #5d View Map
Introduction: The liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous measurement of multiple nicotine metabolites can provide accurate information on smoking status and smoke exposure. However, current SPE or LLE for sample preparation is time consuming and labor intensive. Therefore, we focused on developing a simple and high-throughput method for measuring five smoking-related metabolites (nicotine, cotinine, 3-OH cotinine, nornicotine and anabasine) in population-based urine samples. |
Poster #3c View Map
Introduction |
Poster #27b View Map
Introduction: Early detection of diseases by newborn screening (NBS) is necessary for correct and timely clinical decisions. One of the early methods for NBS of phenylketonuria (PKU) was a fluorimetric method for quantification of phenylalanine (Phe). This method is still used in many countries in south-eastern Europe. The introduction of expanded NBS with tandem mass spectrometry (MS/MS) enabled screening for many diseases, including PKU. Slovenia is now in a unique position to compare the methods because it screens for PKU with a fluorimetric method detecting Phe and also using MS/MS for expanded NBS, also measuring Phe and tyrosine. |
Poster #21h View Map
Aim of the work. Ebola virus disease shows a very high death rate (up to 90%) and the 2014-16 outbreak has been one of the deadliest since 1976, year in which ebola virus was identified. Nevertheless, up to now, any effective pharmacological treatment has been discovered. Some molecules are under study and, among all, remdesivir (RDV) revealed really promising and is now on fase II/III studies. Unfortunately, detailed information about RDV pharmacokinetics are still lacking and no methods for its quantification in patient’s plasma have been reported in literature. |
Poster #7d View Map
Background and aim. Aldosterone, a mineralcorticoid steroid hormone, plays a central role in regulation of blood pressure. The usual serum or plasma concentration is in the range of pg/mL, which makes the analysis rather challenging. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) offer several advantages over conventional radioimmunoassay (RIA)-based assays in terms of both higher sensitivity and better specificity. |
Poster #2c View Map
Introduction: Zearalenone or F-2 mycotoxin is a heat-stable, potent estrogenic metabolic product of some Fusarium and Gibberella species, found in cereal crops like maize, oats, wheat, rice and barley. Due to its similarity to naturally-occurring estrogens (17β-estradiol), zearalenone and its main metabolites (α-zearalenol, β-zearalenol, zeranol and taleranol) are considered by World Health Organization and European Commission as endocrine-disrupting chemicals and a possible cause of carcinogenesis. |
Poster #2a View Map
INTRODUCTION: The untargeted nature of metabolomics allows measurement of biofluid chemistry related to both endogenous metabolism and host-environment exposures (i.e. the exposome). Comprehensive coverage of chemically diverse metabolites present in human blood products benefits from the use of multiple methods, each oriented toward a small molecule subset generally segregated by polarity and hydrophobicity. Whilst recent developments in LC-MS profiling methodologies have delivered numerous solutions for the analysis of polar molecules (e.g. via HILIC-MS) and complex lipids, the analysis of moderately hydrophobic and amphipathic molecules in blood products (including much of the exposome) by RPC methodology, is complicated by the suppressive effects of lipids on the ionisation of low molecular weight (LMW) metabolites. Efficient and inexpensive solutions are required for the separation of small molecules from the remaining sample matrix fit for large scale and high throughput applications. |
Poster #25d View Map
Introduction |
Poster #28h View Map
INTRODUCTION: Road checks carried out by the police contemplate the collection of a salivary sample, which is analyzed with first level screening methods. These semi-quantitative results must be confirmed with confirmatory techniques, such as liquid chromatography combined with mass spectrometry. The aim of this study was is to validate a method for the quantitative analysis of the major substances of abuse on a salivary sample with LC-MS / MS systems. |
Poster #2d View Map
INTRODUCTION |
Poster #20f View Map
Introduction: To date, the infection by Hepatitis C Virus (HCV) affects more than 130 million patients worldwide and causes hepatic cirrhosis, hepatocarcinoma and liver transplantation. In the recent years, Directly Acting Antivirals (DAAs) greatly improved the effectiveness of anti-HCV treatments. Despite this, some issues concerning patients with comorbidities and polytherapies (eg. drug-drug interactions) are still present. For this reason, measurement of DAAs concentrations in patients plasma could be useful for the pharmacokinetic research or Therapeutic Drug Monitoring (TDM). |
Poster #5b View Map
Introduction: Zika virus (ZIKV) is a flavivirus transmitted by Aedes aegypti mosquito, with high incidence of cases on Americas between 2015 and 2016. Although 80% of the cases are asymptomatic, ZIKV infection was associated to neurological complications such as Guillain-Barré syndrome and microcephaly. Based on these reports several initiatives started to understand viral infection process in human and mosquito cells. During experimental investigations it was demonstrated that ZIKV can impair neuronal growth, reduce neural stem cell and even mediate cell death. A hypothesis about the oncolytic potential of ZIKV due its ability of cell growth impairment was raised and later confirmed by in vitro and in vivo studies with glioblastoma cells. The antiproliferative effect of a ZIKV prototype (ZVp) was tested for several types of tumor cell resulting in ZVp activity against prostate cancer cells (PC-3). However, little is known about the cellular mechanisms associated prostate cancer cell death induced by Zika moieties. |
Poster #23h View Map
Introduction: |
Poster #6b View Map
<b>Introduction</b> |
Poster #21g View Map
Introduction: Looking insight pathological processes, metallothioneins (MTs) are considered to be potential biomarkers for monitoring of a development of various types of diseases, such as cancer. The early identification of the MTs in biological tissues could be important tool for the estimation of appropriate clinical therapy. |
Poster #20i View Map
Introduction |
Poster #15d View Map
Introduction: |
Poster #5c View Map
Introduction: Gangliosides (GSs), sialic acid-containing glycosphingolipids, are essential components of cellular membranes, especially in neuronal cells. Besides affecting the function of the membrane in which they are located, these amphiphilic lipids modulate a variety of biological functions through transmembrane signalling.1 GSs content and composition change during aging, which is the most important risk factor for several neurodegenerative diseases.2 Cerebral organoids are a useful model to study composition of membrane GSs in brain tissue or specific cell populations. The brain tissue abundance of GSs expectedly correlates with circulating levels in peripheral blood and perhaps the determination of GSs in human serum can serve as clinically significant marker. |
Poster #21d View Map
Introduction |
Poster #14c View Map
INTRODUCTION: Secondary electrospray ionization-mass spectrometry (SESI-MS) is a highly sensitive innovative real time technique for analyses of complex gaseous mixtures such as human breath. Accurate quantification of gaseous analytes cannot be currently done without internal standards. Breath is a complex mixture containing several hundreds volatile organic compounds (VOCs) present at concentrations from pptv to ppmv. When compounds are present at elevated concentrations that can significantly decrease the reagent ion signal in SESI, the secondary and ternary reactions take place, additional products are observed and quantification based on calibration is compromised. However, accurate quantification of trace gases in air based on chemical ionisation by reagent ions and ion-molecule reaction kinetics can be achieved by selected ion flow tube mass spectrometry (SIFT-MS) that is less sensitive than SESI-MS. |
Poster #28c View Map
Background: Aldosterone is a mineralocorticoid steroid hormone that plays a key role in the regulation of blood pressure. We developed a LC-MS/MS method for the measurement of plasma aldosterone for clinical research purposes. An analytically sensitive method was developed using a mixed-mode Solid Phase Extraction (SPE) sorbent in 96-well plate format. Automated extraction was employed, enabling high throughput of samples. Samples were injected onto an ACQUITY UPLC™ I-Class system with separation on a Waters™ CORTECS™ C18 2.7µm column with VanGuard™ precolumn using an ammonium fluoride(aq)/methanol gradient. Detection was performed using a Waters Xevo™ TQ-XS mass spectrometer to help quantify very low physiological concentrations of aldosterone. |
Poster #16b View Map
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Poster #22g View Map
Introduction: The global increase in obesity is still one of the major health challenges, which drives the growing interest in understanding factors that influence total energy expenditure. Doubly-labelled water is commonly used to measure total energy expenditure in animals and humans. After a bolus dose of doubly-labelled water, hydrogen leaves the body primarily via water turnover, whereas oxygen is depleted both via water turnover and via CO2-exhalation. The excess disappearance rate of 18O relative to deuterium as a measure for the CO2-production rate is used as indirect measure of total energy expenditure. |
Poster #20h View Map
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Poster #23d View Map
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Poster #27f View Map
Background: Dried Blood Spots (DBS) are an established microsampling technique providing a low-cost approach of collecting, shipping and analyzing samples for clinical research. Ligand-binding assays (LBAs) are often used as the frontline testing methodologies for DBS samples in steroid hormone analysis. Although rapid, the relatively low analytical specificity of the LBAs may necessitate follow-up, using liquid chromatography – tandem mass spectrometry (LC-MS/MS). The challenge is to create a fast, analytically sensitive and selective LC-MS/MS methodology for clinical research. |
Poster #25c View Map
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Poster #9c View Map
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Poster #17a View Map
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Poster #23f View Map
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Poster #24c View Map
INTRODUCTION: The current method of choice for the clinical investigation of analytes in biological matrices is LC coupled to tandem mass spectrometer via electrospray ionization based on the high selectivity and sensitivity it offers. One of the major obstacles with this analytical method is matrix effects, where the ionization of the compounds of interest are altered by co-extracting and co-eluting matrix components. Many mitigation strategies exist on overcoming matrix effects. One of these strategies is the use matrix-matching calibrators, where the matrix of the biological sample and that of the calibration range are the same or similar. Compensation occurs assuming that the calibrator and sample are influenced by matrix components in the same manner. For the analysis of a complex matrix such as urine, where the dilution range is extensive and matrix components are sample specific, finding a matching matrix is challenging. |
Poster #27d View Map
INTRODUCTION: Inborn errors of metabolism related to biosynthesis and remodelling of phospholipids, sphingolipids is a newly emerging area in inherited metabolic disorders. Phospholipids are synthesized by a de-novo process, known as ‘‘Kennedy pathway’’ and are then dispersed asymmetrically by a remodeling process called ''Lands Cycle''.The MBOAT7, subject of the current study, is located in the Lands lipid remodeling pathway and inserts arachidonic acid to the sn-2 position of lysophosphotidylinositol. While there animal models of the MBOAT7 exist to elucidate its functional role, no study has yet been conducted on the effects of the MBOAT7 gene defect in humans. |
Poster #18a View Map
INTRODUCTION: With pandemic emergence and increasing magnitude of flavivirus outbreaks in recent years, there is an urgent need in the development of a robust diagnostic tool for flavivirus diagnosis and typing to better facilitate disease management, surveillance and control. |
Poster #5a View Map
Introduction: Nowadays, surgical treatments for oesophageal and gastric cancer, as well as morbid obesity, are increasingly common and have improved patient life expectancy. However, upper gastro-intestinal (GI) post-operative patients frequently experience a long-term decline of quality of life, manifested by symptoms that have often been associated with small intestinal bacterial overgrowth (SIBO); an increase or change of bacterial population in the gut. The current diagnostic tool used in clinic, hydrogen breath test (HBT), measures the change of concentration of hydrogen in the breath after ingestion of glucose. However, this test has a limited specificity. To alleviate this gap, the quantification of certain volatile organic compounds (VOCs) in the breath can be used to identify SIBO. |
Poster #3a View Map
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Poster #28g View Map
Background: Analysis of plasma catecholamines and metanephrines for clinical research is challenging due to their low concentrations (pg/mL) and the extreme polar nature of the analytes. HILIC LC-MS/MS has been used successfully but can be challenging to implement. To address this issue, we have developed a reversed-phase UPLC-MS/MS method that employs a previously developed mixed-mode SPE procedure and combines it with reversed-phase chromatography using the Waters UPLC™ HSS-PFP column. |
Poster #4d View Map
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Poster #11f View Map
INTRODUCTION: Quick identification of pathogens is crucial for efficiently fighting against infectious diseases. Mass spectrometry is a powerful and discriminative tool for this. Whole-cell MALDI-TOF is successful for most pathogens but requires a pure sample usually obtained after cultivation. Tandem mass spectrometry proteotyping has been shown to be an interesting alternative for complex samples and could avoid the cultivation step. |
Poster #21f View Map
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Poster #11d View Map
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Poster #19c View Map
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Poster #16a View Map
Introduction: Prednisolone and its prodrug, prednisone, are two glucocorticoids banned “in competition” by the World Anti-Doping Agency (WADA) when administered by intravenous, intramuscular or rectal routes. They could be found in human urine even if no exogenous administration occurred, because of an ex vivo Δ1-steroid-hydrogenation of cortisol and cortisone physiologically excreted in urine. WADA has established that an additional confirmatory analysis should be performed on samples in which the prednisone and prednisolone concentrations are between 30 and 60 ng/mL to discriminate their exogenous origin from the ex vivo endogenous one. |
Poster #1a View Map
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Poster #11a View Map
Introduction: The two major metabolically active oestrogens in the non-pregnant population are oestrone (E1) and oestradiol (E2). Accurate measurement of E2 at low concentrations is important in a variety of clinical situations including inborn errors of metabolism, disorders of puberty, post-menopausal women, monitoring aromatase inhibitor treatment, and in men. Measuring oestrogens at very low concentrations remains an analytical challenge. Many published methods involve complex and time-consuming sample preparations, large sample volumes and chemical derivatisation. Here we present a simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to measure serum E1 and E2 in the low pmol/L range. |
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Poster #14a View Map
INTRODUCTION: Over the last decade, liquid chromatography coupled to mass spectrometry (LC-MS) has gained widespread popularity and transitioned from research to routine clinical laboratories. While most clinical research laboratories use triple quadrupole mass spectrometers combined with liquid chromatography (LC-MS/MS), for targeted, selective and sensitive quantitation, an increasing number of laboratories prefer one single MS that can efficiently perform routine quantitation of analytes and also screening, confirm, and characterize unknown analytes (ranging from small to large sizes). As a result we observe increasing demands for ready-to-use complete kits for small molecules running on high resolution instruments. |
Poster #27a View Map
INTRODUCTION: Prostate cancer (PCa) is the most commonly diagnosed genital cancer in men worldwide. In 2017, the American Cancer Society reported a large increase in the rate of new PCa cases, accounting for 57% of all new genital cancer cases. Among patients who develop advanced PCa, 80% suffer from bone metastasis, with a sharp drop in survival rate. |
Poster #28d View Map
Introduction: The N-glycosylation of proteins is one of the most important post-translational modifications relevant to various cancers. Among them, alpha-fetoprotein (AFP) is a widely used serological marker that has been associated with hepatocellular carcinoma (HCC). Even though the level of AFP is increased in serum of HCC patients, its diagnostic sensitivity of HCC is poor because AFP levels are also increased in liver diseases, such as cirrhosis. Therefore, various assays for HCC have been developed to increase the sensitivity and selectivity for diagnosis. Above all, changes of fucosylation have been reported to be associated with the development of HCC. |
Poster #23g View Map
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Poster #15b View Map
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Poster #1b View Map
INTRODUCTION: Brain tumours have a very poor prognosis with 10-year survival currently at 14%. The extent of tumour excision is positively correlated with outcomes, however current neuro-navigation methods offer limited information at the tumour margin. Rapid Evaporative Ionisation Mass Spectrometry (REIMS) coupled with electrosurgery known as Intelligent Knife (iKnife) has been previously shown to provide near real-time tissue characterisation with accuracy, comparable to histopathology in different tumour types. Mass spectrometry imaging (MSI) methods provide the means to detect molecules in a spatially resolved manner and can provide insights into tumour metabolism. Laser Desorption Imaging – Rapid Evaporative Ionisation Mass Spectrometry (LDI-REIMS) is one of such MSI techniques that allow for analysis in ambient conditions. |
Poster #18b View Map
INTRODUCTION: Antihypertensives (AHT) is a class of drugs that are used to treat hypertension. The compliance to antihypertensive therapy is critical to achieve adequate blood pressure and avoid complications such as stroke or myocardial infarction. Therapeutic drug monitoring (TDM) is a reliable approach to assess AHT compliance, moreover TDM helps to distinguish non-compliance from patients with true resistant hypertension. |
Poster #20m View Map
NTRODUCTION: The field of clinical proteomics is rapidly advancing and new mass spectrometry technology as well as streamlined sample preparation workflows are making the use of mass spectrometry in the clinical lab ever more attractive. |
Poster #18c View Map
INTRODUCTION: Glutaric aciduria type 1 (GA-1; OMIM#231670) is an autosomal recessive inborn error of metabolism caused by deficiency of glutaryl-CoA dehydrogenase (GCDH) located in the catabolic pathways of L-lysine, L-hydroxylysine, and L-tryptophan. The enzymatic defect gives rise to neurotoxic metabolite glutaric acid (GA) and 3- hydroxyglutaric acid |
Poster #12a View Map
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Poster #11b View Map
Background: Obesity amongst women of reproductive age is increasingly common in developed nations and has been shown to adversely affect childhood cardio-metabolic, respiratory and cognitive-related health outcomes in offspring. Metabolomic signatures of obesity are readily captured in biofluids samples and could potentially provide a molecular barometer for monitoring excessive gestational weight gain (GWG) and postpartum weight loss (WL) in overweight/obese pregnant women. |
Poster #13c View Map
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Poster #21b View Map
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Poster #22e View Map
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Poster #26e View Map
INTRODUCTION: With the advances of tandem mass spectrometry (MS), it is now easier to identify inborn errors of metabolism (IEM) for clinical research. However, there are some practical challenges in processing and interpreting MS data, for example, 1) an increasing number of targets for research, 2) the complexity, time, and effort of data interpretation, and 3) the lack of hands-on experience due to the occurrence of rare diseases. The success of an IEM workflow depends largely on how it processes, interprets, and integrates data and information from multiple sources. Automated reporting and interpretation tools are solutions which could streamline or expand the capability of data processing software for efficient data review, and rapid report delivery. |
Poster #7c View Map
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Poster #14g View Map
INTRODUCTION: Hyperandrogenism and oligomenorrhea are common traits of patients with polycistic ovary syndrome (POCS) and nonclassic congenital andrenal hyperplasia (NCAH). As these disorders have a similar presentation, their distinction using solely clinical symptoms is difficult. Of steroid hormones, 17-hydroxyprogesterone concentration is commonly used as discriminator between POCS and NCAH, although the universal cut off value is not yet established. |
Poster #8b View Map
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Poster #20b View Map
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Poster #22f View Map
Introduction: Hypertension is a major preventable cause of morbidity and mortality which affects one in four adults. Despite this, high rates of non-adherence to antihypertensive medications have been reported both in the U.K. and worldwide. Poor adherence to medications results in wasted resources, sub optimal treatment and worse health outcomes for patients. Objective measurements of adherence to treatment for hypertension such as LC-MS/MS measurement of drugs in urine have been proven to help identify these patients and allow therapy to be adapted. This has in turn improved patient outcomes. In NHS Lothian in Edinburgh, an initial pilot audit revealed that only 64% of patients with resistant hypertension adhered to their treatment regimen. This prompted the development of a local LC-MS/MS method to measure antihypertensive drugs in urine. |
Poster #12c View Map
Background: Therapeutic drug monitoring of isavuconazole, which is a novel broad spectrum antimycoticum against invasive fungal infections, ensures an effective exposure of the drug and minimizes the risk of toxicity (1). The aim of the present study was to evaluate a widely applicable LCMS method eligible for the clinical routine use for the quantification of isavuconaole. |
Poster #20e View Map
INTRODUCTION: Shotgun proteomics is a high throughput method to identify peptides and proteins from complex biological samples. However, it is not an unequivocal procedure, due to the variability of LC-MS/MS experimental conditions and the search engine parameters used in data analysis. Further, in case of data-dependent approach, there may be significant run-to-run variability in the peaks selected for MS/MS analysis resulting in different set of identified peptides even from identical LC-MS/MS runs. |
Poster #19f View Map
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Poster #7b View Map
Introduction: The species belonging to the Enterobacter genus are responsible for 5 to 10% of infections among patients hospitalized in intensive care units and are primarily due to the members of the Enterobacter cloacae complex (ECC). This complex is divided into 13 clusters with 6 defined species E. asburiae, E. cloacae, E. hormaechei, E. kobei, E. ludwigii and E. nimipressuralis. |
Poster #6d View Map
INTRODUCTION: Steroid profiling is of clinical significance for the diagnosis of a wide variety of diseases as steroids play a major role in the regulation of several physiological functions. Historically, these analytes have been measured using immunoassays. However, it is now established that these methods suffer from lack of specificity, due to cross-reactivity. Mass spectrometry has become a standard for accurate steroid profiling, taking advantage of its specificity, high sensitivity and the ease of sample preparation. Nevertheless, due to their small size and their large range of polarity, the analysis by LC-MS/MS is still a challenge. |
Poster #21e View Map
INTRODUCTION: A fundamental assumption in quantitative mass spectrometry is that the internal standard compound behaves identical on all stages of the analytic process chain (ionization-fragmentation-detection) in relation to the target compound and is so used to compensate any inconsistency during sample preparation and analysis. Differential impact of matrix effects on these processes can potentially affect accuracy and sensitivity. Consequently, it can be of significance to investigate the fragmentation characteristics of compounds comparatively (analyte vs. internal standard compound) and under various matrix conditions (reflecting different approaches to sample preparation) in addition to the usual investigated ion source phenomena. The evaluation of such experiments can be performed by breakdown curves, which can be described as a function of the collision energy. |
Poster #16c View Map
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Poster #26g View Map
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Poster #28f View Map
Introduction: Type 2 Diabetes (T2D), the most prevalent form of diabetes, is a metabolic disorder characterized by decreased insulin sensitivity and abnormal hepatic glucose production. Monitoring metabolic alterations during T2D progression may provide better understanding of its pathogenesis and identify potential biomarkers for early diagnosis. Several metabolomics approaches have been applied in diabetic research for identification of metabolites associated with the risk of T2D and related pathways. Here, a semi-targeted workflow was designed to confidently measure known metabolic differentiators, such as branched-chain amino acids, while allowing for the discovery of previously unidentified metabolites that are altered during T2D progression. This approach combines high resolution accurate mass Orbitrap™ technology for maximum detection of known and unknown metabolites in serum samples, with intelligence-driven fragmentation for the identification of knowns and structural elucidation of unknown biomarkers. |
Poster #12d View Map
INTRODUCTION Parathyroid hormone (PTH) is a common clinical marker whose quantification relies on immunoassays, giving variable results as batch, brand, or target epitope changes. Moreover, immunoassays may cross-react with PTH variants such as C-terminal fragments stemming from PTH catabolism. These issues make it difficult to compare results obtained in different laboratories. A reference quantification method is necessary to harmonize PTH assays, both sensitive and selective enough to detect PTH at low concentrations among a variety of closely related compounds. |
Poster #6c View Map
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Poster #23b View Map
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Poster #19a View Map
INTRODUCTION: Metabolomics data often contain thousands of features, but only some of them keep useful information about clinical status and other types of system biology source of data. The one of the first step to the realization of global concepts (such as personalized medicine and system biology) is design a list of the most stable and robust approaches to the extraction of informative metabolites. |
Poster #18d View Map
Introduction: Sepsis remains the leading cause of mortality in the intensive care units. Peripheral blood monocytes have central role in sepsis and are source of numerous effector molecules as well as diagnostic or prognostic markers. Volatile substances (VOCs) from serum samples might be used as reliable markers for detection of important metabolism changes related to innate cells exuberant activation. Therefore development of analytical methods for monocyte VOCs detection and identification is crucial for further characterization of monocyte metabolism alterations. |
Poster #14b View Map
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Poster #23c View Map
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Poster #15a View Map
INTRODUCTION: Recently, MS-based proteomics has become a powerful tool in protein identification and characterization. Particularly for applications in a clinical setting where key information is to be obtained from samples of high complexity and variability, keeping up the pace with the ever-growing requirements is only possible via fully exploiting the potential in our instrumentation and data analysis workflows. In this respect, we recognized a lack of studies directly targeted toward achieving optimum proteomics identification and protein sequence coverage in bottom-up experiments. |
Poster #19d View Map
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Poster #17c View Map
INTRODUCTION: Vitamin D plays a key role in metabolic processes in human body. Despite the growing social awareness, its large deficit is still being observed, which is particularly important in the case of pregnant women. Vitamin D concentration in the fetal period strictly depends on the maternal concentration. Deficiency of this vitamin in the newborn affects a number of diseases, including abnormal development of the bone or immune system. For this reason, preventing deficiency from the first days of life is extremely important. |
Poster #23a View Map
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Poster #19e View Map
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Poster #2b View Map
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Poster #11e View Map
OBJECTIVE: Many detailed guidelines for LC-MS method development are available and well known in the literature. However, the description of the method itself is hardly standardized. Particularly in the field of LC-MS, many variables like different manufacturers and day-to-day or even hour-to-hour variability of instruments have to be considered. |
Poster #9b View Map
INTRODUCTION: Polyunsaturated fatty acids (PUFAs) play essential roles in human physiology. Changes in PUFAs metabolism may have implications in obesity, type 2 diabetes, insulin resistance, cardiovascular diseases, and also interrelationship with other metabolic pathways. Considering their importance, accurate quantification of n-3 and n-6 PUFAs is required. |
Poster #25a View Map
INTRODUCTION: Carcinoid tumours are rare neoplasms arising from the enterochromaffin cells of the neuroendocrine system. Midgut carcinoids are often associated with hyper-secretion of serotonin and can result in carcinoid syndrome; with symptoms including wheezing, diarrhoea and flushing. Measurement of urinary excretion of serotonin metabolite 5-HIAA is widely recommended for both the diagnosis and monitoring of midgut carcinoid tumours. However, collection of 24-hour urine samples can be burdensome, error prone and subject to dietary and medication induced fluctuations. Substitution with a single plasma sample would prove more convenient for patients. Combined measurement of related compounds may also allow a greater understanding of tryptophan metabolism in the carcinoid patient. AIMS: Development of a LC-MS/MS assay for the simultaneous quantification of plasma serotonin, its precursor tryptophan, 5-HIAA and associated compound kynurenine in accordance with CLSI C62A guidelines and establishment of healthy population reference ranges for all analytes. METHOD: Following protein precipitation, chromatographic separation of analytes and their respective deuterated internal standards was achieved following a 10µL injection onto a C18 Fortis column (4.6 x 150mm, 5.0µm). Analysis was carried out using a Waters Acquity® UPLC system coupled to a Xevo TQD® tandem mass spectrometer operating in ESI-positive mode. Multiple reaction monitoring was used to detect analyte and corresponding internal standard transitions. Matrix effects together with assay linearity, imprecision, accuracy, recovery, stability and sensitivity were assessed. Sample comparisons for both 5-HIAA (n=38) and tryptophan (n=12) were carried out using local established assays. |
Poster #26a View Map
Introduction. In last few years multi-target drugs have gained high popularity at the drug development market. Its main pharmacological effect is provided by the combined action of hybrid compounds that interact with several targets in the area of one disease. Novel biogenic molecules, prostanit® and nitroproston®, represent an interesting example of multi-target compounds. They are based on natural prostaglandins PGE1 (in prostanit®) and PGE2 (in nitroproston®) linked by a glycerol moiety to two nitric oxide (NO) ‒ donating fragments. Due to biogenic nature of the these pharmaceuticals, as well as rapid integration of their active components into biochemical cycles, metabolism study becomes difficult. To overcome these complexities, metabolomic approaches were used, giving an opportunity to investigate their metabolic pathways, mechanisms of action and therapeutic effectiveness. |
Poster #22d View Map
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Poster #6a View Map
INTRODUCTION: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in Europe in both men and women. Discovering less invasive diagnostic tools is essential for clinical practice, in particular for the early stage detection, but also for patient stratification. Previous studies showed significant changes of total plasma N-glycome in CRC. Assessing the site-specific glycosylation of an isolated carrier protein can provide valuable insights into CRC-associated glycosylation signatures. Previously, it was reported that elevated difucosylation of haptoglobin (Hp) tetraantennary glycans can be used to discriminate between early stage of hepatocellular carcinoma and cirrhosis. Hp is an acute-phase protein which is synthesized in the liver and is one of the major serum glycoproteins. We here present the first large-scale study of Hp glycosylation in CRC. |
Poster #15c View Map
Introduction: Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has been widely adopted as an analytically sensitive and selective technique for measuring estrone and estradiol in complex matrices such as human blood plasma or serum. Quantitation of these steroids down to low-picogram per milliliter levels are required by many clinical researchers. We endeavored to achieve this using a triple-stage quadrupole mass spectrometer with the most efficient production, isolation and transmission of precursor ions and fastest selective reaction monitoring. |
Poster #28e View Map
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Poster #16d View Map
Introduction: Human breath is a complex mixture containing endogenous metabolites and exogenous compounds. Carboxylic acids represents an interesting class of metabolites. There are several reports about volatile organic acids related to various diseases; such as hexanoic acid in gastro-esophageal cancer, propanoic acid in lung cancer [1], unsaturated fatty acids in irritable bowel disease and acetic acid in cystic fibrosis [2] with concentration ranging from pptv to ppbv. Secondary electrospray ionization mass spectrometry (SESI-MS) is a highly sensitive real time technique for online analyses of trace amounts of VOCs in exhaled breath. |
Poster #22a View Map
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Poster #14f View Map
Introduction: Venetoclax, a BCL-2-inhibitor for the treatment of B-cell lymphoma associated with chromosome 17p deletion, is used more and more in the clinics. Because of severe toxicity, venetoclax has to be titrated at the beginning of the therapy. Further, it is metabolized over CYP3A4 and therefore prone to interactions, which makes therapeutic drug monitoring highly recommended. |
Poster #19b View Map
Introduction: Toxins or poisons are substances which already in small doses may lead to malfunction, damage to health or death. Prompt toxicological analysis may allow early identification of the toxic agent and give therapeutic guidance. In clinical laboratories, screening methods by immunoassays using polyclonal antibodies are commonly applied. Antibody-based screening, however, may cause a relevant number of false positive and false negative results. Detected drugs are not quantified. Furthermore, immunological screening requires a set of 5 to 10 different assays. Nonetheless, comprehensive drug screening and quantification of relevant substances by chromatography with mass spectrometry is often indispensable. |
Poster #3d View Map
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Poster #20c View Map
Introduction:Transplantation is the treatment associated with increased survival rate and greater quality of patient’s life when compared to conventional dialysis. Even nowadays transplantology suffers from the lack of reliable methods of organ quality assessment. The standard protocols are limited to macroscopic appearance inspection or invasive tissue biopsy which do not provide a comprehensive information about the graft. Kidney is the organ largely associated with metabolic processes, thus measurements of metabolites concentrations may permit determining potential organ quality biomarkers and predicting the graft outcome. Hence, there is a need for new diagnostic solution allowing on site graft monitoring and quick decision-making processes during the surgery. |
Poster #14i View Map
Introduction: The stability of mRNA largely depends on the 3’- end and 5’- end structures which are targets for enzymatic degradation. Chemical modifications of those regions are introduced into in vitro transcribed mRNAs in order to confer on them resistance to degradation and increase their translation efficiency, thereby increasing their therapeutic potential. Chemically modified mRNA is a promising therapeutic agent with potential uses in cancer immunotherapy and gene replacement therapy. |
Poster #13e View Map
INTRODUCTION: The analysis of glycoproteins in biological samples is a challenging task. Routine glycoproteomics approaches involve sample enrichment, proteolytic digestion followed by de-glycosylation, the analysis of components with HPLC-MS/MS and data assessment via database search. However, during de-glycosylation the site-specificity of the information is lost, and during DDA MS/MS analyses many minor components are overlooked. |
Poster #26c View Map
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Poster #26b View Map
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Poster #27g View Map
INTRODUCTION: The most dynamic process which regulates DNA methylation is recently discovered active demethylation. It involves ten-eleven translocation (TET) enzymes to catalyze stepwise oxidation of 5-methylcytosine (5-mCyt) to 5-hydroxymethylcytosine (5-hmCyt) and further demethylation products 5-formylcytosine (5-fCyt) and 5-carboxylcytosine (5-caCyt). Mutations targeting TET genes are frequently observed in acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). Mutations in genes: succinate dehydrogenase (SDH), fumarate hydratase (FH) and isocitrate dehydrogenase (IDH) are also found in acute leukemias. These mutations result in accumulation of the succinate (SA), fumarate (FA), 2-oxoglutarate (2-OG) and R-2-hydroxyglutarate (R-2HG). It may deregulate the activity of TET enzymes and, in turn, DNA demethylation. Although the oncogenic mechanism of these mutations remains still under investigation, determine of these metabolites could be relevant for diagnosis, prognosis and treatment of a subset of patients with AML and MDS. |
Poster #27c View Map
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Poster #10c View Map
Introduction: Metabolic disorders are caused by the accumulation of metabolites in the body which lead to irreversible physiological effects. The first-tier screening procedure for these disorders is typically performed on dried blood spot (DBS) samples by the MS/MS system. When a newborn screen is found to be positive, second-tier tests using plasma samples are provided to reduce false-positive and false-negative results. To prevent resampling and provide comfort for newborns, it is more convenient to use the same DBS sample taken in the first stage of screening. Objective: The objective of this study was development of a method based on the DBS instead of plasma analyses for each aminoacidemia disorder that could potentially use as second-tier tests. Methods: To optimize the extraction conditions of 19 naturally occurring amino acids from DBS, the central composite design and response surface methodology were used to demonstrate the influence of effective factors on the responses. Also, four different types of validation were compared, and the best validation method was selected. |
Poster #8d View Map
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Poster #14d View Map
INTRODUCTION: Fluoropyrimidines (5-fluorouracil or capecitabine) are anticancer drugs that can lead to severe or lethal toxicities in case of dihydropyrimidine dehydrogenase (DPD) deficiency. In France, health authorities recommend the determination of uracil concentration to guide dosing of fluoropyrimidines. Numerous LC-MS/MS methods have been proposed but they include complex liquid-liquid or solid-phase extraction procedures. |
Poster #20d View Map
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Poster #28b View Map
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Poster #4a View Map
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Poster #20j View Map
INTRODUCTION: Correct measurement of biogenic monoamines is essential in the clinical diagnosis and follow-up of pheochromocytoma, paraganglioma and carcinoid tumours. Currently we measure the catecholamines (adrenaline (A), noradrenaline (NA), dopamine(D)), their metabolites (vanillylmandelic acid (VMA) and homovanillic acid (HVA)) and the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) with two different fluorometric methods, while the metanephrines (metanephrine (MN), normetanephrine (NMN), 3-methoxytyramine (3MT) are measured with an electrochemical detection method. These three methods lack analytical specificity and sensitivity and are time consuming. |
Poster #24a View Map
Desmosine is a promising biomarker for estimating elastin degradation activity in diseases like chronic obstructive pulmonary disease and cystic fibrosis and for monitoring the effect of therapeutic interventions. We successfully developed an LC-MS/MS assay for the measurement of desmosine in urine and plasma, but encountered numerous obstacles during the developmental and validation process. |
Poster #26f View Map
Introduction: Since treatment advances for lysosomal storage disorders (LSDs), the application of mass spectrometry (MS) techniques have expanded to screening for some of the treatable LSDs. To date, flow injection MS (FI-MS) is generally the preferred screening technique to be of diagnostic value for 6 LSDs, namely Pompe, MPS-I, Fabry, Gaucher, Niemann-Pick A/B and Krabbe disorders, from a single dried blood spot (DBS) sample. We evaluated the analytical performance and diagnostic precision of a 6-plex LSD enzyme assay utilizing the technique of liquid chromatography with tandem MS (LC-MS/MS) within the clinical laboratory setting. |
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Poster #13f View Map
Introduction: Treatment of advanced cutaneous melanoma remains challenging, and new data on melanoma biology are required. The most widely accepted criteria for the prognostic evaluation of melanoma are histopathological and clinical parameters, and the identification of additional tumor markers is thus of paramount importance. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI), an important tool in cancer research, is useful for unraveling the molecular profile of melanoma. |
Poster #11c View Map
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Poster #21c View Map
Introduction:Transplantology is one of the fastest growing area of medicine. However, it faces many serious problems, such as lack of effective tools for organ quality assessment. Since tissue biopsy is invasive and macroscopic visual assessment is unreliable, new technologies are strongly needed. A lipidomic approach is a possibility to better understanding the mechanism of reperfusion grafts injury and identify potential organ quality biomarkers. |
Poster #3b View Map
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Poster #22b View Map
INTRODUCTION: Phospholipids are major constituents of plasma membranes and have been shown to cause severe ion suppression or enhancement in the analysis of biological samples by liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS). Additionally, phospholipids tend to accumulate on reversed-phase columns, causing a decrease in column performance and reduced life time. |
Poster #23e View Map
INTRODUCTION: Plasma metanephrines are recommended as one of the first line biochemical tests for the diagnosis of phaeochromocytomas and paragangliomas. LC-MS/MS methods are typically less susceptible to analytical interference than other methods, such as HPLC. However, analytical interference from midodrine, a sympathomimetic, and metformin, has been reported in LC-MS/MS assays for plasma metanephrines, affecting the ability to accurately quantitate metanephrine. We report an LC-MS/MS method for plasma metanephrines that has been demonstrated to be free of interference from midodrine, its metabolite, desglymidodrine, and from metformin. |
Poster #8a View Map
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Poster #26d View Map
Background |
Poster #27h View Map
INTRODUCTION: Congenital adrenal hyperplasia (CAH), is a disorder which affects the function of adrenal glands in producing one or more of the steroid hormones. The commonest causes of CAH are the 21-hydroxylase deficiency (95%) and other rarer forms such as 11-beta hydroxylase deficiency. The analysis of 17-hydroxyprogesterone, androstenedione and cortisol is the best screening test for CAH, while the analysis of corticosterone, 11-deoxycortisol, 21-deoxycortisol, 11-deoxycorticosterone, progesterone and testosterone can either be the adjunct to evaluation of 21-hydroxylase deficiency or the evaluation of defects of adrenal steroid biosynthesis. |
Poster #9d View Map
Aim: To identify the serum metabolomics signature that is correlated with the chemoradiosensitivity of esophageal squamous cell carcinoma (ESCC). |
Poster #17b View Map
Introduction : Peptide sequencing in mass spectrometry generally followed through tandem mass spectrometry with the collision-induced dissociation (CID) process, which uses accelerated noble gases for fragmentation. However, peptide sequencing with CID process is inadequate due to the requirement of expensive instrument and expert technicians. Recently titanium oxide (TiO2) nanowires for laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS) was introduced with less background noises on low mass region. Moreover, its photocatalytic activity with UV due to the high band gap energy takes attention as a multifunctional solid matrix system. |
Poster #13d View Map
INTRODUCTION: Most clinical laboratories measure total 25-hydroxy vitamin D by immunoassays with variable analytical performance. Other relevant metabolites, such as 25-hydroxy vitamin D2 (25(OH)D2), 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) and 25,26-dihydroxyvitamin D3 (25,26(OH)2D3) may provide additional information beyond the simple measurement of 25(OH)D. Liquid-chromatography tandem mass-spectrometry (LC-MS/MS) allows the simultaneous measurement of multiple vitamin D metabolites with high sensitivity and specificity. |
Poster #28a View Map
Introduction: Bioavailable testosterone (BioT) includes free and albumin-bound testosterone. Total testosterone (TT) can be measured by immunoassays and LC-MS/MS, a preferred method for women and children. BioT can be calculated using various algorithms based on TT, SHBG, and albumin concentrations or measured after removal of SHBG-bound testosterone by NH4SO4 precipitation. We compared BioT results generated by different in-house approaches with a reference lab method. |
Poster #22c View Map
Introduction: Liver transplant surgery is currently the standard of treatment in patients with end-stage organ failure. Nowadays, the dominant method of organ preservation used by most transplantation centers is the static cold storage (SCS). However, a better method of organ preservation is sought, which would allow extending the storage time of the graft while maintaining its proper quality The proposed method is normothermic ex-vivo liver perfusion (NEVLP), based on maintaining normal metabolic activity, which gives the opportunity to better assessment of liver viability before implantation. One of the possibilities is to assess the production of bile by the liver perfused in these conditions. It is considered that the production of bile alone is not sufficient evidence for the proper functioning of the liver and directs the research to assess the composition of bile. Therefore, it is assumed that changes in the concentration of bile acids, which are the main component of bile, may correlate with changes occurring in the transplanted organ. |
