Efficacy of a multicomponent binding agent against combined exposure to zearalenone and ochratoxin A in weaned pigs.

Introduction: The study aimed to evaluate the efficacy of a novel multicomponent substance against combined exposure to the mycotoxins zearalenone (ZEN) and ochratoxin A (OTA) in weaned piglets. Methods: In total, 60 piglets at the age of 28 days were equally allocated to four experimental groups (A-D), consisting of eight female and seven male piglets each (15 animals per group, for a total trial duration of 42 days). Animals from group A received typical weaner feed without mycotoxins or the test product [multicomponent mycotoxin detoxifying agent (MMDA)]. Group B animals received the same weaner feed contaminated with 0.992 mg ZEN/kg feed and 0.531 mg OTA/kg feed without the addition of the MMDA. Animals in group C received the same contaminated feed as group B with the addition of 1.5 g MMDA/kg feed, whereas group D received the same feed as group B with the inclusion of 3 g MMDA/kg feed. Clinical signs and performance parameters [body weight (BW), average daily weight gain (ADWG), and feed conversion ratio (FCR)] were evaluated, while mycotoxin residues were also assessed in the liver and kidney tissues. Results: Findings showed improved FCR in the group that received the greatest dose of the test product (3 g MMDA/kg feed) compared to the group that received the lower dose (1.5 g MMDA/kg feed). A few hematological and biochemical parameters were slightly altered, predominantly within normal limits. The residue analysis demonstrated a reduction of OTA in liver samples, a-ZEL in the liver and total tested samples, and a total of ZEN and metabolite contents in all samples of the group that received the greatest MMDA dose in comparison to the group that received the toxins without the addition of the test product. Discussion: Therefore, a positive effect of the MMDA at the greatest dosage regime on reducing bioavailability and tissue deposition of ZEN and OTA, with a particularly positive effect on FCR in weaned pigs, is suggested under concurrent ZEN and OTA exposure in vivo.

Comparison of chromatographic conditions for the targeted tandem mass spectrometric determination of 354 mammalian metabolites.

Metabolomics is becoming increasingly popular in livestock research, but no single analytical method can cover the entire metabolome. As such, we compared similar and complementary chromatographic methods with respect to analyte coverage and chromatographic properties of mammalian metabolites. We investigated 354 biologically relevant primary metabolites from 19 compound classes including amino acids, bile acids, biogenic amines, carboxylic acids, lipids, nucleotides and sugars. A total of 2063 selected reaction monitoring transitions were optimized on a triple quadrupole mass spectrometer. We then determined the retention profiles and peak parameters of our compounds using an anion exchange chromatography (AIC), three reversed-phase (RP) and three hydrophilic interaction liquid chromatography (HILIC) methods. On average, HILIC methods covered 54% of all metabolites with retention factors >1, while average RP coverage was 41%. In contrast to RP, HILIC methods could also retain polar metabolites such as amino acids and biogenic amines. Carboxylic acids, nucleotides, and sugar related compounds were best separated by AIC or zwitterionic pHILIC with alkaline eluents. Combining two complementary HILIC and RP methods increased the library coverage to 92%. By further including important short chain fatty acids, a combination of HILIC, RP and AIC methods achieved a coverage of 97%. The resulting dataset of LC and MS/MS parameters will facilitate the development of tailor-made quantitative targeted LC-MS/MS methods to investigate the mammalian metabolome.

The Beneficial Effect of the Mobile Application Euglyca in Children and Adolescents with Type 1 Diabetes Mellitus: A Randomized Controlled Trial.

Background: Euglyca® is a mobile application which we developed for children and adolescents suffering type 1 diabetes mellitus (T1DM) for calculation of the appropriate insulin bolus dose by importing in the equation carbohydrates, lipids, glucose levels, and personalized parameters. Aim of this study is to evaluate the efficacy of this application on patients' glycemic control and satisfaction. Subjects and Methods: Eighty children and adolescents (aged 13.5 ± 2.8 years old, mean ± standard deviation) with T1DM were included in the study and were randomly and equally assigned in two groups. Patients were asked to use Euglyca for the calculation of the bolus insulin dose in the E group and to pursue their routine calculations in the C group (controls). At baseline and at 3, 6, and 12 months following the initial visit, glycated hemoglobin (HbA1c) values, percentages of hypoglycemias, hyperglycemias, and normoglycemias were determined for each patient, while Diabetes Treatment Satisfaction Questionnaire (DTSQ) was used to assess patients' treatment satisfaction at baseline and at 6 and 12 months. Results: Children and adolescents in the E group had a statistically significant decrease in HbA1c values and increase in percentages of normoglycemias and DTSQ scores, in comparison to children and adolescents in the C group. In the E group, a statistically significant positive linear correlation was found between DTSQ score and percentages of normoglycemias and a statistically significant negative correlation between changes in percentages of normoglycemias (Δnormoglycemias) and changes in HbA1c (ΔHbA1c). Conclusions: The use of the mobile application Euglyca contributes to the improvement of glycemic control and treatment satisfaction of children and adolescents with T1DM.

The role of inter-species interactions in Salinispora specialized metabolism.

Bacterial genome sequences consistently contain many more biosynthetic gene clusters encoding specialized metabolites than predicted by the compounds discovered from the respective strains. One hypothesis invoked to explain the cryptic nature of these gene clusters is that standard laboratory conditions do not provide the environmental cues needed to trigger gene expression. A potential source of such cues is other members of the bacterial community, which are logical targets for competitive interactions. In this study, we examined the effects of such interactions on specialized metabolism in the marine actinomycete Salinispora tropica. The results show that antibiotic activities and the concentration of some small molecules increase in the presence of co-occurring bacterial strains relative to monocultures. Some increases in antibiotic activity could be linked to nutrient depletion by the competitor as opposed to the production of a chemical cue. Other increases were correlated with the production of specific compounds by S. tropica. In particular, one interaction with a Vibrio sp. consistently induced antibiotic activity and was associated with parent ions that were unique to this interaction, although the associated compound could not be identified. This study provides insight into the metabolomic complexities of bacterial interactions and baseline information for future genome mining efforts.

Brucella Seroprevalence in a High-Risk Population in Greece: A Cross-Sectional Study.

INTRODUCTION: Brucellosis is a zoonosis with high occupational risk. However, seroprevalence of Brucella antibodies among occupational groups is not known, since studies in endemic countries are rare. METHODS: A cross-sectional seroprevalence study was conducted among livestock farmers in an endemic region in Greece. A low-risk group of individuals that just moved in the region was used as controls. Rose Bengal, Wright standard tube agglutination (STA) tests, and specific IgG and IgM antibodies using ELISA were evaluated; differences and odds ratios were calculated. Results were compared with studies from other endemic regions. RESULTS: 100 livestock farmers and family members and 34 first-year students were enrolled. Rose Bengal results were 18% positive versus 0% (p=0.007); Wright STAs for Brucella melitensis were 8% versus 2.9% (p=0.448) and for Brucella abortus they were 2% versus 2.9% (p=0.588). ELISA IgG was positive in 8% of farmers versus 2.9% of students (p=0.448). Parallel testing with Rose Bengal and ELISA IgG was positive in 3% versus 0% (p=0.571). No significant odds ratios were calculated for Wright STAs and ELISA IgG. CONCLUSIONS: Healthy livestock farmers may present with positive Rose Bengal test but this translates to true seroprevalence in only a small proportion. Livestock farmers have no significant seroprevalence that may obscure diagnosis of acute brucellosis in clinical settings.

Mass Spectrometry Based Molecular 3D-Cartography of Plant Metabolites.

Plants play an essential part in global carbon fixing through photosynthesis and are the primary food and energy source for humans. Understanding them thoroughly is therefore of highest interest for humanity. Advances in DNA and RNA sequencing and in protein and metabolite analysis allow the systematic description of plant composition at the molecular level. With imaging mass spectrometry, we can now add a spatial level, typically in the micrometer-to-centimeter range, to their compositions, essential for a detailed molecular understanding. Here we present an LC-MS based approach for 3D plant imaging, which is scalable and allows the analysis of entire plants. We applied this approach in a case study to pepper and tomato plants. Together with MS/MS spectra library matching and spectral networking, this non-targeted workflow provides the highest sensitivity and selectivity for the molecular annotations and imaging of plants, laying the foundation for studies of plant metabolism and plant-environment interactions.

Prioritizing Natural Product Diversity in a Collection of 146 Bacterial Strains Based on Growth and Extraction Protocols.

In order to expedite the rapid and efficient discovery and isolation of novel specialized metabolites, while minimizing the waste of resources on rediscovery of known compounds, it is crucial to develop efficient approaches for strain prioritization, rapid dereplication, and the assessment of favored cultivation and extraction conditions. Herein we interrogated bacterial strains by systematically evaluating cultivation and extraction parameters with LC-MS/MS analysis and subsequent dereplication through the Global Natural Product Social Molecular Networking (GNPS) platform. The developed method is fast, requiring minimal time and sample material, and is compatible with high-throughput extract analysis, thereby streamlining strain prioritization and evaluation of culturing parameters. With this approach, we analyzed 146 marine Salinispora and Streptomyces strains that were grown and extracted using multiple different protocols. In total, 603 samples were analyzed, generating approximately 1.8 million mass spectra. We constructed a comprehensive molecular network and identified 15 molecular families of diverse natural products and their analogues. The size and breadth of this network shows statistically supported trends in molecular diversity when comparing growth and extraction conditions. The network provides an extensive survey of the biosynthetic capacity of the strain collection and a method to compare strains based on the variety and novelty of their metabolites. This approach allows us to quickly identify patterns in metabolite production that can be linked to taxonomy, culture conditions, and extraction methods, as well as informing the most valuable growth and extraction conditions.

Natural products as mediators of disease.

Covering: up to 2016Humans are walking microbial ecosystems, each harboring a complex microbiome with the genetic potential to produce a vast array of natural products. Recent sequencing data suggest that our microbial inhabitants are critical for maintaining overall health. Shifts in microbial communities have been correlated to a number of diseases including infections, inflammation, cancer, and neurological disorders. Some of these clinically and diagnostically relevant phenotypes are a result of the presence of small molecules, yet we know remarkably little about their contributions to the health of individuals. Here, we review microbe-derived natural products as mediators of human disease.

Mass Spectrometry-Based Visualization of Molecules Associated with Human Habitats.

The cars we drive, the homes we live in, the restaurants we visit, and the laboratories and offices we work in are all a part of the modern human habitat. Remarkably, little is known about the diversity of chemicals present in these environments and to what degree molecules from our bodies influence the built environment that surrounds us and vice versa. We therefore set out to visualize the chemical diversity of five built human habitats together with their occupants, to provide a snapshot of the various molecules to which humans are exposed on a daily basis. The molecular inventory was obtained through untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of samples from each human habitat and from the people that occupy those habitats. Mapping MS-derived data onto 3D models of the environments showed that frequently touched surfaces, such as handles (e.g., door, bicycle), resemble the molecular fingerprint of the human skin more closely than other surfaces that are less frequently in direct contact with humans (e.g., wall, bicycle frame). Approximately 50% of the MS/MS spectra detected were shared between people and the environment. Personal care products, plasticizers, cleaning supplies, food, food additives, and even medications that were found to be a part of the human habitat. The annotations indicate that significant transfer of chemicals takes place between us and our built environment. The workflows applied here will lay the foundation for future studies of molecular distributions in medical, forensic, architectural, space exploration, and environmental applications.

Sharing and community curation of mass spectrometry data with Global Natural Products Social Molecular Networking.

The potential of the diverse chemistries present in natural products (NP) for biotechnology and medicine remains untapped because NP databases are not searchable with raw data and the NP community has no way to share data other than in published papers. Although mass spectrometry (MS) techniques are well-suited to high-throughput characterization of NP, there is a pressing need for an infrastructure to enable sharing and curation of data. We present Global Natural Products Social Molecular Networking (GNPS; http://gnps.ucsd.edu), an open-access knowledge base for community-wide organization and sharing of raw, processed or identified tandem mass (MS/MS) spectrometry data. In GNPS, crowdsourced curation of freely available community-wide reference MS libraries will underpin improved annotations. Data-driven social-networking should facilitate identification of spectra and foster collaborations. We also introduce the concept of 'living data' through continuous reanalysis of deposited data.

A metabolomics guided exploration of marine natural product chemical space.

INTRODUCTION: Natural products from culture collections have enormous impact in advancing discovery programs for metabolites of biotechnological importance. These discovery efforts rely on the metabolomic characterization of strain collections. OBJECTIVE: Many emerging approaches compare metabolomic profiles of such collections, but few enable the analysis and prioritization of thousands of samples from diverse organisms while delivering chemistry specific read outs. METHOD: In this work we utilize untargeted LC-MS/MS based metabolomics together with molecular networking to. RESULT: This approach annotated 76 molecular families (a spectral match rate of 28 %), including clinically and biotechnologically important molecules such as valinomycin, actinomycin D, and desferrioxamine E. Targeting a molecular family produced primarily by one microorganism led to the isolation and structure elucidation of two new molecules designated maridric acids A and B. CONCLUSION: Molecular networking guided exploration of large culture collections allows for rapid dereplication of know molecules and can highlight producers of uniques metabolites. These methods, together with large culture collections and growing databases, allow for data driven strain prioritization with a focus on novel chemistries.

Outcome of patients with acute myocardial infarction admitted in hospitals with or without catheterization laboratory: results from the HELIOS registry.

AIMS: To compare the treatment and outcomes of myocardial infarction patients in hospitals with and without catheterization laboratory. METHODS AND RESULTS: The Hellenic Infarction Observation Study was a countrywide registry of acute myocardial infarction, conducted during 2005-2006. The registry enrolled 1840 patients with myocardial infarction from 31 hospitals with a proportional representation of all types of hospitals and of all geographical areas. Out of these patients, 645 (35%) were admitted in 11 hospitals with and 1195 (65%) in 20 hospitals without catheterization laboratory. Patients admitted in hospitals with catheterization laboratory in comparison with patients admitted in hospitals without were younger (66+/-14 vs. 68+/-13, P<0.004) with less diabetes (27 vs. 33%, P<0.001), but without other baseline differences (female 27 vs. 25%, prior myocardial infarction 20 vs. 17%, Killip class>1 22 vs. 23%). Reperfusion rates for ST-segment elevation myocardial infarction were 67% (43% lytic, 24% primary percutaneous coronary interventions) versus 56% (55% lytic, 1% percutaneous coronary interventions; P<0.01). In-hospital outcomes in hospitals with versus in hospitals without laboratory were: mortality 6.5 versus 8.3% (NS), stroke 2.2 versus 1.1% (NS), major bleeding 1.1 versus 0.6% (NS), and heart failure 11 versus 16% (P<0.01). In multivariate regression analysis, being admitted in a hospital without catheterization laboratory was not an independent predictor of increased in-hospital mortality (odds ratio=1.18, 95% confidence interval: 0.72-1.93, P=0.505). CONCLUSION: Although the majority of acute myocardial infarction patients was admitted in hospitals without catheterization laboratory, these patients do not have a survival disadvantage, provided they are treated with lytic therapy, medical secondary prevention drugs, and eventual revascularization according to current guidelines.

Relation of C-reactive protein to the first onset and the recurrence rate in lone atrial fibrillation.

The presence of systemic inflammation determined by elevations in high-sensitivity C-reactive protein (hs-CRP) has been associated with persistence of atrial fibrillation (AF). The influence of inflammation markers, such as hs-CRP, on the recurrences of lone AF, however, has not been clarified. We tested the hypothesis of whether, in patients with a first paroxysmal episode of lone AF, the hs-CRP levels were elevated, and whether elevated hs-CRP could predict the recurrence rate of lone AF in patients without antiarrhythmic drugs. Using a case-control study design, the hs-CRP levels in 125 patients with a documented symptomatic first paroxysmal episode of lone AF was compared with the hs-CRP levels in 65 control patients. hs-CRP levels are presented as median values with the interquartile range (25th to 75th percentiles). The hazard ratio compared the 75th percentile of hs-CRP with the 25th percentile. In the arrhythmia group, hs-CRP was higher than in the control patients (median 0.23 mg/dl, interquartile range 0.12 to 0.49, vs 0.087 mg/dl, interquartile range 0.058 to 0.098, p <0.001). After adjusting for baseline characteristics, including, age, gender, and baseline blood pressure, hs-CRP remained a significant predictor of recurrent AF (hazard ratio 1.15, 95% confidence interval 1.04 to 1.24, p = 0.002) at 2 years of follow-up. In conclusion, this study is the first to document that the first paroxysmal episode of lone AF is associated with elevated hs-CRP levels, suggesting that hs-CRP may be a marker for inflammatory states that may promote the initiation of lone AF. These pathways may represent a novel mechanism by which structural changes resulting from inflammation could induce lone AF. The elevated hs-CRP levels could also predict the recurrence rate of lone AF in patients without antiarrhythmic drugs.

Effect of radiofrequency catheter ablation on the biochemical marker ischemia modified albumin.

Ischemia-modified albumin (IMA) levels were measured after radiofrequency (RF) catheter ablation to evaluate the effect of direct myocardial necrosis on IMA formation. IMA levels have been shown to increase in patients after RF catheter ablation compared with those who undergo diagnostic electrophysiologic studies. The results of this study suggest that IMA may be a marker of myocardial injury.