Silicon's Influence on Polyphenol and Flavonoid Profiles in Pea (Pisum sativum L.) under Cadmium Exposure in Hydroponics: A Study of Metabolomics, Extraction Efficacy, and Antimicrobial Properties of Extracts.

The current study aimed to investigate the impact of silicon (Si) supplementation in the form of Na2SiO3 on the metabolome of peas under normal conditions and following exposure to cadmium (Cd) stress. Si is known for its ability to enhance stress tolerance in various plant species, including the mitigation of heavy metal toxicity. Cd, a significant contaminant, poses risks to both human health and the environment. The study focused on analyzing the levels of bioactive compounds in different plant parts, including the shoot, root, and pod, to understand the influence of Si supplementation on their biosynthesis. Metabolomic analysis of pea samples was conducted using a targeted HPLC/MS approach, enabling the identification of 15 metabolites comprising 9 flavonoids and 6 phenolic acids. Among the detected compounds, flavonoids, such as flavon and quercetin, along with phenolic acids, including chlorogenic acid and salicylic acid, were found in significant quantities. The study compared Si supplementation at concentrations of 1 and 2 mM, as well as Cd stress conditions, to evaluate their effects on the metabolomic profile. Additionally, the study explored the extraction efficiency of three different methods: accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and maceration (MAC). The results revealed that SFE was the most efficient method for extracting polyphenolic compounds from the pea samples. Moreover, the study investigated the stability of polyphenolic compounds under different pH conditions, ranging from 4.0 to 6.0, providing insights into the influence of the pH on the extraction and stability of bioactive compounds.

Metabolic profiling of bacteria with the application of polypyrrole-MOF SPME fibers and plasmonic nanostructured LDI-MS substrates.

Here we present application of innovative lab-made analytical devices such as plasmonic silver nanostructured substrates and polypyrrole-MOF solid-phase microextraction fibers for metabolic profiling of bacteria. For the first time, comprehensive metabolic profiling of both volatile and non-volatile low-molecular weight compounds in eight bacterial strains was carried out with utilization of lab-made devices. Profiles of low molecular weight metabolites were analyzed for similarities and differences using principal component analysis, hierarchical cluster analysis and random forest algorithm. The results showed clear differentiation between Gram positive (G+) and Gram negative (G-) species which were identified as distinct clusters according to their volatile metabolites. In case of non-volatile metabolites, differentiation between G+ and G- species and clustering for all eight species were observed for the chloroform fraction of the Bligh & Dyer extract, while methanolic fraction failed to recover specific ions in the profile. Furthermore, the results showed correlation between volatile and non-volatile metabolites, which suggests that lab-made devices presented in the current study might be complementary and therefore, useful for species differentiation and gaining insights into bacterial metabolic pathways.

Silver Lactoferrin as Antimicrobials: Mechanisms of Action and Resistance Assessed by Bacterial Molecular Profiles.

A diverse silver-lactoferrin (AgLTF) complex, comprising silver ions (Ag+) and silver nanoparticles, displayed a synergistic antibacterial effect while being almost five times more lethal than LTF alone. Gas chromatography-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-in linear (LP) and reflectron (RP) positive modes-were used to comprehensively analyze metabolites and proteins profiles of bacteria (Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA) and Enterococcus faecalis (EF)) treated using AgLTF complex versus exclusively Ag+. Although both agents resulted in similar metabolic shifts in bacteria, AgLTF significantly triggered the production of sulfides (related to bacterial stress resistance), ethanol, 2-butanol (indicating exhaustion of cell respiration), decanoic acid, and nonane (suggesting ongoing oxidative stress). Keto acids formation and fermentation pathways were enhanced by AgLTF and suppressed by Ag+. Furthermore, AgLTF appears to interact with proteins fraction of bacteria in a concentration-dependent manner. EF molecular profiles showed less changes between treated and untreated bacteria. On the other hand, SA and PA proteins and metabolic patterns were the most differentiated from untreated bacteria. In conclusion, our study may provide valuable insights regarding the molecular mechanisms involved in AgLTF antimicrobial action.

Lipidomics Characterization of the Microbiome in People with Diabetic Foot Infection Using MALDI-TOF MS.

Lipidomic profiling has emerged as a powerful tool for the comprehensive characterization of bacterial species, particularly in the context of clinical diagnostics. Utilizing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), this study aims to elucidate the lipidomic landscapes of bacterial strains isolated from diabetic foot infections (DFI). Our analysis successfully identified a diverse array of lipids in the cellular membranes of both Gram-positive and Gram-negative bacteria, revealing a total of 108 unique fatty acid combinations. Specifically, we identified 26 LPG, 33 LPE, 43 PE, 114 PG, 89 TAG, and 120 CLP in Gram-positive bacteria and 10 LPG, 14 LPE, 124 PE, 37 PG, 13 TAG, and 22 CLP in Gram-negative strains. Key fatty acids, such as palmitic acid, palmitoleic acid, stearic acid, and oleic acid, were prominently featured. Univariate analysis further highlighted distinct lipidomic signatures among the bacterial strains, revealing elevated levels of phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) in Gram-negative bacteria associated with DFI. In contrast, Gram-positive strains demonstrated increased or uniquely fluctuating levels of triglyceride (TAG) and cardiolipin (CLP). These findings not only underscore the utility of MALDI-TOF MS in bacterial lipidomics but also provide valuable insights into the lipidomic adaptations of bacteria in diabetic foot infections, thereby laying the groundwork for future studies aimed at constructing microbial lipid libraries for enhanced bacterial identification.

Alternative matrices in forensic toxicology: a critical review.

PURPOSE: The use of alternative matrices in toxicological analyses has been on the rise in clinical and forensic settings. Specimens alternative to blood and urine are useful in providing additional information regarding drug exposure and analytical benefits. The goal of this paper is to present a critical review on the most recent literature regarding the application of six common alternative matrices, i.e., oral fluid, hair, sweat, meconium, breast milk and vitreous humor in forensic toxicology. METHODS: The recent literature have been searched and reviewed for the characteristics, advantages and limitations of oral fluid, hair, sweat, meconium, breast milk and vitreous humor and its applications in the analysis of traditional drugs of abuse and novel psychoactive substances (NPS). RESULTS: This paper outlines the properties of six biological matrices that have been used in forensic analyses, as alternatives to whole blood and urine specimens. Each of this matrix has benefits in regards to sampling, extraction, detection window, typical drug levels and other aspects. However, theses matrices have also limitations such as limited incorporation of drugs (according to physical-chemical properties), impossibility to correlate the concentrations for effects, low levels of xenobiotics and ultimately the need for more sensitive analysis. For more traditional drugs of abuse (e.g., cocaine and amphetamines), there are already data available on the detection in alternative matrices. However, data on the determination of emerging drugs such as the NPS in alternative biological matrices are more limited. CONCLUSIONS: Alternative biological fluids are important specimens in forensic toxicology. These matrices have been increasingly reported over the years, and this dynamic will probably continue in the future, especially considering their inherent advantages and the possibility to be used when blood or urine are unavailable. However, one should be aware that these matrices have limitations and particular properties, and the findings obtained from the analysis of these specimens may vary according to the type of matrix. As a potential perspective in forensic toxicology, the topic of alternative matrices will be continuously explored, especially emphasizing NPS.

A New Approach to Imaging and Rapid Microbiome Identification for Prostate Cancer Patients Undergoing Radiotherapy.

(1) Background: Little is known about the impact of urinary microflora, in particular, its effects on side effects after radiotherapy. The use of mass spectrometry identification method (MALDI) may bring a new look at the issue of the composition and significance of the urinary microbiome. This study aimed to use the mass spectrometry identification method (MALDI) to identify the microbiome of urine samples collected from 50 irradiated prostate cancer patients. (2) Methods: Blood and urine samples were collected before gold marker implantation, at the start and last day of radiotherapy, 1, 4 months after. Patients do not always collect the urine from the midstream; therefore, samples were collected from the first void and midstream in 12 patients for MALDI analysis; in the remaining 38 patients-from the midstream void for MALDI and biochemical analysis. (3) Results: Microorganisms were present in 140/181 urine samples. We found 33 different species 3G(-) and 30G(+). The most frequently isolated strains were: Staphylococcus haemolyticus, Staphylococcus epidermidis, Staphylococcus hominis, Enterococcus faecalis, and Micrococcus luteus. When comparing the type of urine samples, bacteria were more common in samples from the first-void urine than from the midstream one. The absence of bacteria was found in 12.2% of samples from the first-void urine and in 24.7% from the midstream. There was no difference in the total incidence of species between streams (p = 0.85). Before fiducial implantation, the total number of detected bacterial species was significantly higher in comparison to the end of radiotherapy (p = 0.038), indicating that the administered therapy resulted in depleting the local microbiome. One month after radiotherapy, an increase in the number of isolated bacteria was observed. The number of bacterial species in urine did not correlate with blood parameters. The presence of leukocytes (p = 0.013) and proteins (p = 0.004) in urine was related to a greater variety of bacteria found in urine specimens. (4) Conclusions: We obtained a similar spectrum of bacteria from the initial and middle urine streams. We also showed that there is a change in bacteria species affected by the treatment of prostate cancer patients, with both antibiotics before gold fiducial implantation and radiotherapy.

Lipid Constituents of Diatoms (Halamphora) as Components for Production of Lipid Nanoparticles.

Lipid nanocarriers smaller than 200 nm may be used as pharmaceutical/cosmetic raw materials as they are able to penetrate the skin. The nanostructured lipid carriers (NLCs) based on microalgae oil (Schizochytrium) and lipids extracted from diatoms (Halamphora cf. salinicola (strain SZCZM1454A)) were produced by the HSH (high shear homogenization) method. Fatty acid profile of crude oil from diatoms indicated the presence of palmitoleic, palmitic, stearic acid, oleic and myristic acids as the most common fatty acids in the strain investigated. The quantitative composition and the synthesis condition of NLC dispersions were optimized by using the full factorial designs. The physicochemical parameters of the obtained lipid nanocarriers were characterized by SEM, DSC and XRD measurements and the fraction with the optimum parameters (size below 200 nm, polydispersity index not exceeding 0.2 and zeta potential higher than +45 mV) was selected for further study. The positive charge of the obtained lipid nanoparticles is beneficial as permits electrostatic bonding with the negatively charged skin surface. As follows from stability tests, the NLCs obtained could be stored at room temperature.

Citric Acid as a Potential Prostate Cancer Biomarker Determined in Various Biological Samples.

Despite numerous studies, the molecular mechanism of prostate cancer development is still unknown. Recent investigations indicated that citric acid and lipids-with a special emphasis on fatty acids, steroids and hormones (ex. prolactin)-play a significant role in prostate cancer development and progression. However, citric acid is assumed to be a potential biomarker of prostate cancer, due to which, the diagnosis at an early stage of the disease could be possible. For this reason, the main goal of this study is to determine the citric acid concentration in three different matrices. To the best of our knowledge, this is the first time for citric acid to be determined in three different matrices (tissue, urine and blood). Samples were collected from patients diagnosed with prostate cancer and from a selected control group (individuals with benign prostatic hyperplasia). The analyses were performed using the rapid fluorometric test. The obtained results were correlated with both the histopathological data (the Gleason scale as well as the Classification of Malignant Tumors (pTNM) staging scale) and the biochemical data (the values of the following factors: prostate specific antigen, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglyceride, total cholesterol, creatinine and prolactin) using chemometric methods. For tissue samples, the results indicated a decreased level of citric acid in the case of prostate cancer. The analyte average concentrations in serum and urine appeared to be corresponding and superior in the positive cohort. This trend was statistically significant in the case of urinary citric acid. Moreover, a significant negative correlation was demonstrated between the concentration of citric acid and the tumor stage. A negative correlation between the total cholesterol and high-density lipoprotein and prolactin was particularly prominent in cancer cases. Conversely, a negative association between low-density lipoprotein and prolactin levels was observed solely in the control group. On the basis of the results, one may assume the influence of hormones, particularly prolactin, on the development of prostate cancer. The present research allowed us to verify the possibility of using citric acid as a potential biomarker for prostate cancer.

Comparison Study of Cytotoxicity of Bare and Functionalized Zinc Oxide Nanoparticles.

In this paper, a study of the cytotoxicity of bare and functionalized zinc oxide nanoparticles (ZnO NPs) is presented. The functionalized ZnO NPs were obtained by various types of biological methods including microbiological (intra- and extracellular with Lactobacillus paracasei strain), phytochemical (Medicago sativa plant extract) and biochemical (ovalbumin from egg white protein) synthesis. As a control, the bare ZnO NPs gained by chemical synthesis (commercially available) were tested. The cytotoxicity was measured through the use of (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) dye as well as lactate dehydrogenase (LDH) assays against murine fibroblast L929 and Caco-2 cell lines. As a complementary method, scanning electron microscopy (SEM) was performed to assess the morphology of the tested cells after treatment with ZnO NPs. The microscopic data confirmed the occurrence of apoptotic blebbing and loss of membrane permeability after the administration of all ZnO NPs. The reactive oxygen species (ROS) concentration during the cell lines' exposure to ZnO NPs was measured fluorometrically. Additionally, the photocatalytic degradation of methylene blue (MB) dye in the different light conditions, as well as the antioxidant activity of bare and functionalized ZnO NPs, is also reported. The addition of all types of tested ZnO NPs to methylene blue resulted in enhanced rates of photo-degradation in the presence of both types of irradiation, but the application of UV light resulted in higher photocatalytic activity of ZnO NPs. Furthermore, bare (chemically synthetized) NPs have been recognized as the strongest photocatalysts. In the context of the obtained results, a mechanism underlying the toxicity of bio-ZnO NPs, including (a) the generation of reactive oxygen species and (b) the induction of apoptosis, is proposed.

Development of controlled film of polypyrrole for solid-phase microextraction fiber by electropolymerization.

The aim of the study was to develop an approach for synthesis of polypyrrole coating material and further fabrication of solid-phase microextraction fiber by direct electropolymerization on a stainless-steel wire. Surface morphology, porosity and thermal stability were evaluated by different physico-chemical methods. Synthesized polypyrrole fiber was successfully applied for extraction of VOCs such as benzene, toluene, ethylbenzene, p-xylene, phenol and dodecane. Utilization of polypyrrole fiber for extraction of VOCs was assessed by HS-SPME coupled to gas chromatography with flame ionization detection (GC-FID). Solid-phase microextraction parameters for their simultaneous analysis were optimized using Box-Behnken design and accounted for 49.7 min for extraction time, 30 °C for extraction temperature and 17.2 min for equilibration time. In addition, the coating showed good reproducibility (RSD < 12.22%), while the values of limit of detection were in the range of 0.59-283.33 ng/mL.

Metabolic Profiling of VOCs Emitted by Bacteria Isolated from Pressure Ulcers and Treated with Different Concentrations of Bio-AgNPs.

Considering the advent of antibiotic resistance, the study of bacterial metabolic behavior stimulated by novel antimicrobial agents becomes a relevant tool to elucidate involved adaptive pathways. Profiling of volatile metabolites was performed to monitor alterations of bacterial metabolism induced by biosynthesized silver nanoparticles (bio-AgNPs). Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae and Proteus mirabilis were isolated from pressure ulcers, and their cultures were prepared in the presence/absence of bio-AgNPs at 12.5, 25 and 50 µg mL-1. Headspace solid phase microextraction associated to gas chromatography-mass spectrometry was the employed analytical platform. At the lower concentration level, the agent promoted positive modulation of products of fermentation routes and bioactive volatiles, indicating an attempt of bacteria to adapt to an ongoing suppression of cellular respiration. Augmented response of aldehydes and other possible products of lipid oxidative cleavage was noticed for increasing levels of bio-AgNPs. The greatest concentration of agent caused a reduction of 44 to 80% in the variety of compounds found in the control samples. Pathway analysis indicated overall inhibition of amino acids and fatty acids routes. The present assessment may provide a deeper understanding of molecular mechanisms of bio-AgNPs and how the metabolic response of bacteria is untangled.

Needle Trap Device-GC-MS for Characterization of Lung Diseases Based on Breath VOC Profiles.

Volatile organic compounds (VOCs) have been assessed in breath samples as possible indicators of diseases. The present study aimed to quantify 29 VOCs (previously reported as potential biomarkers of lung diseases) in breath samples collected from controls and individuals with lung cancer, chronic obstructive pulmonary disease and asthma. Besides that, global VOC profiles were investigated. A needle trap device (NTD) was used as pre-concentration technique, associated to gas chromatography-mass spectrometry (GC-MS) analysis. Univariate and multivariate approaches were applied to assess VOC distributions according to the studied diseases. Limits of quantitation ranged from 0.003 to 6.21 ppbv and calculated relative standard deviations did not exceed 10%. At least 15 of the quantified targets presented themselves as discriminating features. A random forest (RF) method was performed in order to classify enrolled conditions according to VOCs' latent patterns, considering VOCs responses in global profiles. The developed model was based on 12 discriminating features and provided overall balanced accuracy of 85.7%. Ultimately, multinomial logistic regression (MLR) analysis was conducted using the concentration of the nine most discriminative targets (2-propanol, 3-methylpentane, (E)-ocimene, limonene, m-cymene, benzonitrile, undecane, terpineol, phenol) as input and provided an average overall accuracy of 95.5% for multiclass prediction.

Lipidomics as a Diagnostic Tool for Prostate Cancer.

The main goal of this study was to explore the phospholipid alterations associated with the development of prostate cancer (PCa) using two imaging methods: matrix-assisted laser desorption ionization with time-of-flight mass spectrometer (MALDI-TOF/MS), and electrospray ionization with triple quadrupole mass spectrometer (ESI-QqQ/MS). For this purpose, samples of PCa tissue (n = 40) were evaluated in comparison to the controls (n = 40). As a result, few classes of compounds, namely phosphatidylcholines (PCs), lysophosphatidylcholines (LPCs), sphingomyelins (SMs), and phosphatidylethanolamines (PEs), were determined. The obtained results were evaluated by univariate (Mann-Whitney U-test) and multivariate statistical analysis (principal component analysis, correlation analysis, volcano plot, artificial neural network, and random forest algorithm), in order to select the most discriminative features and to search for the relationships between the responses of these groups of substances, also in terms of the used analytical technique. Based on previous literature and our results, it can be assumed that PCa is linked with both the synthesis of fatty acids and lipid oxidation. Among the compounds, phospholipids, namely PC 16:0/16:1, PC 16:0/18:2, PC 18:0/22:5, PC 18:1/18:2, PC 18:1/20:0, PC 18:1/20:4, and SM d18:1/24:0, were assigned as metabolites with the best discriminative power for the tested groups. Based on the results, lipidomics can be found as alternative diagnostic tool for CaP diagnosis.

Cigarette smoke exposure causes systemic and autonomic cardiocirculatory changes in rats depending on the daily exposure dose.

AIMS: To evaluate the systemic changes and autonomic cardiocirculatory control of awaken rats chronically exposed to the cigarette smoke (CS) of 1 or 2 cigarettes/day. MAIN METHODS: Rats were exposed to clean air (control) or cigarette smoke of 1 (CS1) or 2 (CS2) cigarettes/animal/day for 30 days. Then, arterial pressure (AP) and heart rate (HR) were recorded in conscious rats to assess spontaneous baroreflex sensitivity and HR and AP variabilities. Evoked baroreflex and cardiac autonomic tone were evaluated by vasoactive drugs and autonomic blockers, respectively. In another group, ventilatory and cardiovascular parameters were recorded under hypoxia and hypercapnia stimulus. At the end of protocols, heart, lung, kidneys and liver were collected for histological analysis. KEY FINDINGS: Rats exposed to CS showed morphological changes, being more evident in the CS2 group. Also, less weight gain and cardiac hypertrophy were prominent in CS2 rats. Basal AP and HR, spontaneous baroreflex sensitivity and cardiovascular variabilities were similar among groups. CS exposure progressively blunted the bradycardia response to phenylephrine (-2.2 ± 0.1 vs. -1.7 ± 0.2 vs. -1.5 ± 0.2) while the tachycardia response to sodium nitroprusside was slightly increased compared to control. Vagal tone was not affected by CS, but CS2 rats exhibited higher sympathetic tone (-25 ± 4 vs. -28 ± 4 vs. -56 ± 9) and lower intrinsic HR (411 ± 4 vs. 420 ± 8 vs. 390 ± 6). Exposure to CS of 2 cigarettes also exacerbated the reflex cardiovascular and ventilatory responses to hypoxia and hypercapnia. SIGNIFICANCE: CS exposure for 30 days promoted systemic changes and autonomic cardiocirculatory dysfunction in rats depending on the daily exposure dose.

Evaluation of salivary VOC profile composition directed towards oral cancer and oral lesion assessment.

OBJECTIVES: Endogenous substances have been analyzed in biological samples in order to be related with metabolic dysfunctions and diseases. The study aimed to investigate profiles of volatile organic compounds (VOCs) from fresh and incubated saliva donated by healthy controls, individuals with oral tissue lesions and with oral cancer, in order to assess case-specific biomarkers of oxidative stress. MATERIALS AND METHODS: VOCs were pre-concentrated using headspace-solid phase microextraction and analyzed using gas chromatography-mass spectrometry. Then, VOCs positively modulated by incubation process were subtracted, yielding profiles with selected features. Principal component analysis and hierarchical cluster analysis were used to inspect data distribution, while univariate statistics was applied to indicate potential markers of oral cancer. Machine learning algorithm was implemented, aiming multiclass prediction. RESULTS: The removal of bacterial contribution to VOC profiles allowed the obtaining of more specific case-related patterns. Artificial neural network model included 9 most relevant compounds (1-octen-3-ol, hexanoic acid, E-2-octenal, heptanoic acid, octanoic acid, E-2-nonenal, nonanoic acid, 2,4-decadienal and 9-undecenoic acid). Model performance was assessed using 10-fold cross validation and receiver operating characteristic curves. Obtained overall accuracy was 90%. Oral cancer cases were predicted with 100% of sensitivity and specificity. CONCLUSIONS: The selected VOCs were ascribed to lipid oxidation mechanism and presented potential to differentiate oral cancer from other inflammatory conditions. CLINICAL RELEVANCE: These results highlight the importance of interpretation of saliva composition and the clinical value of salivary VOCs. Elucidated metabolic alterations have the potential to aid the early detection of oral cancer and the monitoring of oral lesions.

Evolution and Evaluation of GC Columns.

A chromatographic column is the fundamental element required for gas-chromatographic analysis. The separation of components coming from complex mixtures, prior to their detection was leading to a prominent revolution in different areas of science. Moreover, current advances in gas chromatographic (GC) columns technology and development have been providing almost unlimited possibilities for analysis employing diverse matrices. We aim through this review article to describe the evolution of chromatographic columns, by pointing the most important stages, as well as the new trends and future perspectives predicted for the new generation of GC columns. Furthermore, it was in our scope to present the main fundamentals regarding the theoretical relationships that describe the chromatographic separation, to introduce concepts related to columns selection in accordance with the required application as well as to discuss the available evaluation parameters for columns efficiency. Consequently, the early stages of first columns preparation up to the development of GC capillary columns used nowadays, together with examples of their applications are also reported and described in detail.

A Review of GC-Based Analysis of Non-Invasive Biomarkers of Colorectal Cancer and Related Pathways.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer in the world. In Europe, it is the second most common cause of cancer-related deaths. With the advent of metabolomics approaches, studies regarding the investigation of metabolite profiles related to CRC have been conducted, aiming to serve as a tool for early diagnosis. In order to provide further information about the current status of this field of research, 21 studies were systematically reviewed, regarding their main findings and analytical aspects. A special focus was given to the employment of matrices obtained non-invasively and the use of gas chromatography as the analytical platform. The relationship between the reported volatile and non-volatile biomarkers and CRC-related metabolic alterations was also explored, demonstrating that many of these metabolites are connected with biochemical pathways proven to be involved in carcinogenesis. The most commonly reported CRC indicators were hydrocarbons, aldehydes, amino acids and short-chain fatty acids. These potential biomarkers can be associated with both human and bacterial pathways and the analysis based on such species has the potential to be applied in the clinical practice as a low-cost screening method.

Study on Molecular Profiles of Staphylococcus aureus Strains: Spectrometric Approach.

Staphylococcus aureus remains a major health problem responsible for many epidemic outbreaks. Therefore, the development of efficient and rapid methods for studying molecular profiles of S. aureus strains for its further typing is in high demand. Among many techniques, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS) represents a timely, cost-effective, and reliable strain typing approach, which is still rarely used due to insufficient knowledge about the impact of sample preparation and analysis conditions on the molecular profiles and strain classification efficiency of S. aureus. The aim of this study was to evaluate the effect of the culture conditions and matrix type on the differentiation of molecular profiles of various S. aureus strains via the MALDI TOF MS analysis and different computational methods. The analysis revealed that by changing the culture conditions, matrix type, as well as a statistical method, the differentiation of S. aureus strains can be significantly improved. Therefore, to accelerate the incorporation of the MALDI-based strain typing in routine laboratories, further studies on the standardization and searching of optimal conditions on a larger number of isolates and bacterial species are of great need.

New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS.

A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, cyclitols, and their respective products generated through the electrochemistry (EC) process.

Investigation of Ayahuasca ß-Carboline Alkaloids and Tryptamine in Sweat Samples from Religious Community Participants by GC-MS.

Ayahuasca, a hallucinogenic beverage used in religious rituals in South America, has become a global phenomenon. Its main active components are the ß-carbolines alkaloids, harmine (HRM) and harmaline (HRL), as well as the potent hallucinogen N,N-dimethyltryptamine (DMT). Despite its rising consumption, information regarding possible clinical applications and toxicological effects of ayahuasca is still limited. This study presents the first investigation of the use of sweat for the determination of DMT, HRM and HRL in ayahuasca users during a religious ritual. Sweat is an alternative matrix with advantages over many conventional biological samples, mainly because the collection procedure is non-invasive, easy and simple and samples can be collected without disturbing the religious ritual. In the study, solid-phase extraction was performed under basic conditions. Linearity was observed ranging from 20 to 1500 ng/patch with coefficients of determination (R2) higher than 0.99 for all analytes. The results indicated high selectivity for all investigated analytes, with extraction efficiency exceeding 70%, accuracy ranging from 87.5 to 102.4%, intra-assay precision of 1.85-9.44% and inter-assay precision between 3.34 and 9.85%. The limits of detection were 15 ng/patch for HRM and HRL and 10 ng/patch for DMT. The sweat proved to be a viable option to monitor ayahuasca use.