Gas Chromatography-Mass Spectrometry (GC-MS) Metabolites Analysis in Endometriosis Patients: A Prospective Observational Translational Study.

BACKGROUND: Endometriosis affects women of reproductive age, and its pathogenesis is still unclear. Typically, it overlaps other similar medical and surgical conditions, determining a delay in early diagnosis. Metabolomics allows studying metabolic changes in different physiological or pathological states to discover new potential biomarkers. We used the gas chromatography-mass spectrometer (GC-MS) to explore metabolic alterations in endometriosis to better understand its pathophysiology and find new biomarkers. METHODS: Twenty-two serum samples of patients with symptomatic endometriosis and ten without it were collected and subjected to GC-MS analysis. Multivariate and univariate statistical analyses were performed, followed by pathway analysis. RESULTS: Partial least squares discriminant analysis was performed to determine the differences between the two groups (p = 0.003). Threonic acid, 3-hydroxybutyric acid, and proline increased significantly in endometriosis patients, while alanine and valine decreased. ROC curves were built to test the diagnostic power of metabolites. The pathway analysis identified the synthesis and degradation of ketone bodies and the biosynthesis of phenylalanine, tyrosine, and tryptophan as the most altered pathways. CONCLUSIONS: The metabolomic approach identifies metabolic alterations in women with endometriosis. These findings may improve our understanding of the pathophysiological mechanisms of disease and the discovery of new biomarkers.

Metabolic Profile of Patients with Severe Endometriosis: a Prospective Experimental Study.

Endometriosis is a common disease affecting women in reproductive age. There are several hypotheses on the pathogenesis of this disease. Often, its lesions and symptoms overlap with those of many other medical and surgical conditions, causing a delay in diagnosis. Metabolomics represents a useful diagnostic tool for the study of metabolic changes during a different physiological or pathological status. We used 1H-NMR to explore metabolic alteration in a cohort of patients with endometriosis in order to contribute to a better understanding of the pathophysiology of the disease and to suggest new useful biomarkers. Thirty-seven patients were recruited for the metabolomic analysis: 22 patients affected by symptomatic endometriosis and 15 not affected by it. Their serum samples were collected and analyzed with 1H-NMR. Multivariate statistical analysis was conducted, followed by univariate and pathway analyses. Partial Least Square Discriminant Analysis (PLS-DA) was performed to determine the presence of any differences between the non-endometriosis and endometriosis samples (R2X = 0.596, R2Y = 0.713, Q2 = 0.635, and p < 0.0001). ß-hydroxybutyric acid and glutamine were significantly increased, whereas tryptophan was significantly decreased in the endometriosis patients. ROC curves were built to test the diagnostic power of the metabolites (ß-hydroxybutyric acid: AUC = 0.85 CI = 0.71-0.99; glutamine: AUC = 0.83 CI = 0.68-0.98; tryptophan: AUC = 0.75 CI = 0.54-0.95; ß-hydroxybutyric acid + glutamine + tryptophan AUC = 0.92 CI = 0.81-1). The metabolomic approach enabled the identification of several metabolic alterations occurring in women with endometriosis. These findings may provide new bases for a better understanding of the pathophysiological mechanisms of the disease and for the discovery of new biomarkers. Trial registration number NCT02337816.

Conservative treatment for well-differentiated endometrial cancer: when and why it should be considered in young women.

The aim of this review was to update current knowledge on the conservative treatment of endometrial cancer (EC) based on a literature review. A web-based search in the MEDLINE database was carried out on EC management and treatment. All relevant information has been collected and analysed. Case series were mainly found in the literature search. Conservative treatments were offered to young patients with stage I low-grade endometrioid carcinomas of the endometrium. Different options included high/low dose progestin treatments, hysteroscopic resection of the disease, a levonorgestrel intrauterine device or a combination of various strategies. The overall complete response rate was near 76.5% with a recurrence rate of up to 33.8%. Pregnancy outcomes reached rates of 64.8% for live births. The current clinical outcomes show that conservative treatment aimed at preserving fertility is feasible for stage I endometrial well-differentiated adenocarcinomas in motivated patients under close monitoring.

Biogenic amines and other polar compounds in long aged oxidized Vernaccia di Oristano white wines.

Oxidized white wines are produced by techniques that provide a barrel ageing which can range from a few years to some decades. This step, characterized by the metabolic activity of peculiar strains of Saccharomyces cerevisiae yeast, can affect the chemical composition of these wines and the production of unwanted substances such as biogenic amines. In this study, Vernaccia di Oristano wines from different vintages have been analysed for the first time regarding the content of biogenic amines and amino acids (by HPLC-FLD), and polar compounds (by HPLC-DAD and LC-MS). Furthermore, colour and technological parameters (contents of alcohol, reducing sugars, total and volatile acidities, pH and organic acids) of the wines were also evaluated. Older samples showed dark shades, which may have derived from polyphenols' oxidation while ageing. Some typical ageing products, such as 5-(hydroxymethyl)furfural and hydroxycinnamic acid derivatives were found in larger quantities in these samples, as well as the purinic compound xanthine, which was also detected in relevant concentrations. Additionally, as expected, the average of the main biogenic amines quantified in Vernaccia di Oristano was higher compared to non-oxidized white wines, especially in the older samples. Thus, though this content does not exceed values which spoil the quality of the wine, the monitoring of the winemaking conditions is suggested, to further limit the presence of these undesirable compounds.

Determination of dansylated amino acids and biogenic amines in Cannonau and Vermentino wines by HPLC-FLD.

Free amino acids (AA) and biogenic amines (BA) were quantified for the first time in Cannonau and Vermentino wines, the two most popular "Controlled Designation of Origin" wines from Sardinia (Italy). An analytical method for the simultaneous determination of AA and BA was developed, using selective derivatization with dansyl chloride followed by HPLC with fluorescence detection. Thirty-two compounds were identified in the wines analysed. High levels of AA were found, with proline being the most abundant with average levels of 1244 ± 398 and 1008 ± 281 mg/L in Cannonau and Vermentino wines, respectively. BA were detected at average concentrations <10mg/L, except putrescine which reached 20.5 ± 10.2mg/L in Cannonau wines. Histamine was never detected in any Vermentino wines. γ-Aminobutyric acid, 4-hydroxyproline, glycine, leucine+isoleucine and putrescine proved to be useful for differentiating Cannonau wines from Vermentino wines.

Color evaluation of seventeen European unifloral honey types by means of spectrophotometrically determined CIE L*Cab*h(ab)° chromaticity coordinates.

CIE (Commission Internationale de l'Eclairage) L(*)Cab(*)h(ab)° color coordinates for 305 samples of 17 unifloral honeys types (asphodel, buckwheat, black locust, sweet chestnut, citrus, eucalyptus, Garland thorn, honeydew, heather, lime, mint, rapeseed, sage, strawberry tree, sulla flower, savory and thistle) from different geographic locations in Europe were spectrophotometrically assessed and statistically evaluated. Preliminary separation of unifloral honeys was obtained by means of L(*)-C(ab)(*) color coordination correlation. Hierarchical Cluster Analysis (HCA) revealed an expected segregation of the honeys types according to their chromatic characteristics. Principal Component Analysis (PCA) allowed to obtain a more defined distinction of the 17 unifloral honey types, particularly when using 3D graphics. CIE L(*)C(ab)(*)hab(*) color coordinates were useful for the identification of several honey types. The proposed method represents a simple and efficient procedure that can be used as a basis for the authentication of unifloral honeys worldwide.

Cornflower (Centaurea cyanus L.) honey quality parameters: chromatographic fingerprints, chemical biomarkers, antioxidant capacity and others.

The samples of cornflower (Centaurea cyanus L.) honey from Poland were subjected to ultrasonic solvent extraction applying the mixture of pentane and diethyl ether 1:2v/v (solvent A) as well as dichloromethane (solvent B). The major compounds of the extracts (analysed by GC-MS/GC-FID) were C13 and C9 norisoprenoids. Among them, (E)-3-oxo-retro-α-ionol (2.4-23.9% (solvent A); 3.9-14.4% (solvent B)) and (Z)-3-oxo-retro-α-ionol (3.7-29.9% (solvent A); 8.4-20.4% (solvent B)) were found to be useful as chemical biomarkers of this honey. Other abundant compounds were: methyl syringate (0.0-31.4% (solvent A); 0.0-25.4% (solvent B)) and 3-hydroxy-4-phenylbutan-2-one (1.6-15.8% (solvent A); 5.1-15.1% (solvent B)). HPLC-DAD analysis of the samples revealed lumichrome (4.7-10.0mg/kg), riboflavin (1.9-2.7mg/kg) and phenyllactic acid (112.1-250.5mg/kg) as typical compounds for this honey type. Antioxidant and antiradical properties as well as total phenolic content of the samples were found to be rather moderate by FRAP (ferric reducing antioxidant power), DPPH (1,1-diphenyl-2-picrylhydrazyl radical) and Folin-Ciocalteu assays, respectively. Additionally, CIE L(∗)a(∗)b(∗)C(∗)h chromatic coordinates were evaluated. Colour attributes of cornflower honey were characterised by elevated values of L(∗) and particularly high values of b(∗) and h coordinates, which correspond to medium bright honey with intense yellow colour.

Comparative analysis of Tunisian wild Crataegus azarolus (yellow azarole) and Crataegus monogyna (red azarole) leaf, fruit, and traditionally derived syrup: phenolic profiles and antioxidant and antimicrobial activities of the aqueous-acetone extracts.

Quantitative and qualitative analyses of the yellow and red azarole phenolic extracts prepared from leaf, fruit peel/pulp, and syrup were comparatively investigated. The yellow azarole was found significantly richer in polyphenols than the red-fruit species. Hyperoside was the main phenolic in both yellow and red azarole leaves and only in yellow fruits, whereas procyanidin B2 was the major compound in red fruits. Yellow azarole leaf and fruit peel extracts exhibited the strongest antioxidant activities using DPPH (≈168 and 79 µmol TEAC/g fw, respectively) and FRAP (≈378 and 161 µmol Fe(2+)/g fw, respectively) assays. The highest antibacterial activities were recorded for the yellow azarole leaf and fruit peel extracts, especially against Staphylococcus aureus and Streptococcus faecalis . The low phenolic content of the syrups contrasted with their significant antioxidant and antimicrobial potentials, which were correlated to their hydroxymethylfurfural (HMF) (furan derivative amounts) content.

Radical scavenging activity and LC-MS metabolic profiling of petals, stamens, and flowers of Crocus sativus L.

UNLABELLED: Radical scavenging activities of Crocus sativus petals, stamens and entire flowers, which are waste products in the production of the spice saffron, by employing ABTS radical scavenging method, were determined. At the same time, the metabolic profiles of different extract (obtained by petals, stamens and flowers) were obtained by LC-ESI-IT MS (liquid chromatography coupled with electrospray mass spectrometry equipped with Ion Trap analyser). LC-ESI-MS is a techniques largely used nowadays for qualitative fingerprint of herbal extracts and particularly for phenolic compounds. To compare the different extracts under an analytical point of view a specific method for qualitative LC-MS analysis was developed. The high variety of glycosylated flavonoids found in the metabolic profiles could give value to C. sativus petals, stamens and entire flowers. PRACTICAL APPLICATION: Waste products obtained during saffron production, could represent an interesting source of phenolic compounds, with respect to the high variety of compounds and their free radical scavenging activity.

Riboflavin and lumichrome in Dalmatian sage honey and other unifloral honeys determined by LC-DAD technique.

Riboflavin (vitamin B(2)) and its metabolite lumichrome were quantified in 117 samples from 11 unifloral honeys types (Arbutus unedo L., Asphodelus microcarpus Salzm. et Viv., Citrus spp., Eucalyptus spp., Hedysarum coronarium L., Castanea sativa L. honeydew, Mentha spp., Paliurus spina-christi., Salix spp., Salvia officinalis L., Satureja spp.). The quantification of these two compounds was performed by LC-DAD method which does not require sample purification. The proposed method in our study has low limits of detection and quantification, very good linearity in a large concentration range and very good precision. It allows simultaneous determination of 5-hydroxymethylfurfural (HMF) and known chemical biomarkers of unifloral honeys such as abscisic acid diastereomers, homogentisic acid, methyl syringate and kynurenic acid. No statistical correlation was observed between riboflavin and lumichrome content. Although, the concentration of vitamin B(2) in honey may be too low (<6.1mg/kg) to generate interest in the field of nutrition, the presence of its main metabolite lumichrome may be useful to determine the botanical origin of certain unifloral honeys. In fact, the analysis of 11 unifloral honey types showed that Dalmatian sage (S. officinalis L.) honey is characterised by unusual high levels of lumichrome (20.2±2.6mg/kg). The botanical origin of lumichrome from sage flower was assessed by analysing bee-stomach extracts. Other analytical parameters, such as total phenols, antioxidant and antiradical activities, HMF and diastase activity were studied in Dalmatian sage honey.