Mapping Aspergillus niger Metabolite Biomarkers for In Situ and Early Evaluation of Table Grapes Contamination

The Aspergillus niger exometabolome was recently investigated using advanced gas chromatography in tandem with multivariate analysis, which allowed a metabolite biomarker pattern to be proposed. Microbial metabolomics patterns have gained enormous relevance, mainly due to the amount of information made available, which may be useful in countless processes. One of the great challenges in microbial metabolomics is related to applications in more complex systems of metabolomics information obtained from studies carried out in culture media, as complications may occur due to the dynamic nature of biological systems. Thus, the main objective of this research was to evaluate the applicability of the A. niger metabololite biomarkers pattern for in situ and early evaluation of table grapes contamination, used as study model. A. niger is a ubiquitous fungus responsible for food contamination, being reported as one of the main agents of the black mold disease, a serious post-harvest pathology of table grapes. This work included analysis from 1 day of growth time of pure A. niger cultures, A. niger cultures obtained from previously contaminated grapes, and finally, an in situ solid-phase microextraction (SPME) approach directly on previously contaminated table grapes. Supervised multivariate analysis was performed which revealed that after 1 day of inoculation it was possible to detect A. niger biomarkers, which can be extremely useful in making this type of method possible for the rapid detection of food contamination. The results obtained confirm the potential applicability of the pattern of A. niger biomarkers for early detection of the fungi (after 1 day of contamination), and may be further explored for access food susceptibility to fungi contamination, based on direct analysis of the food item.

Human and livestock faecal biomarkers at the prehistorical encampment site of Ullafelsen in the Fotsch Valley, Stubai Alps, Austria – potential and limitations

The Ullafelsen at 1869 m a.s.l. in the Tyrolean Stubai Alps next to Innsbruck is an important (geo-)archaeological reference site for the Mesolithic period. Buried fireplaces on the Ullafelsen plateau were dated at 10.9–9.5 cal. kyrs BP and demonstrate together with thousands of flint stone artifacts the presence of hunter-gatherers during the Early Holocene. Most recently, we demonstrated the great potential of n-alkane and black carbon biomarkers for contributing to a better understanding of pedogenesis and landscape evolution. In order to study the importance of human and/or animals for occupation of this relevant geoarchaeological site, we carried out steroid and bile acid analyses on two modern faeces samples from cattle and sheep and on 37 soil samples from seven soil profiles at the Ullafelsen. The modern animal faeces show a dominance of 5β-stigmastanol and deoxycholic acid for ruminants (cattle and sheep), which is in agreement with literature data. The OAh horizons, which have accumulated and developed since the Mesolithic, revealed high contents of steroids and bile acids; the E (LL) horizon coinciding with the Mesolithic living floor is characterized by medium contents of steroids and bile acids. By contrast, the subsoil horizons Bh, Bs and BvCv contain low contents of faecal biomarkers indicating that leaching of steroids and bile acids into the podsolic subsoils is not an important factor. Deoxycholic acid is the most abundant bile acid in all soil samples and gives evidence for strong faeces input of ruminants. The steroid and bile acid patterns and ratios indicate a negligible input of human faeces on the Ullafelsen. β-Sitosterol as plant-derived steroid has also a strong influence on the faecal biomarker pattern in our soils. Root input into the subsoils is likely reflected by β-sitosterol contents. In conclusion, our results reflect a strong faecal input by livestock, rather than by humans as found for other Anthrosols such as Amazonian Dark Earths. Further studies need to focus on the question of the exact timing of faeces deposition.

Mapping the Aspergillus niger metabolite biomarkers for in situ and early evaluation of table grapes contamination

Aspergillus niger volatilome was recently explored using advanced gas chromatography tools tandem with multivariate analysis, which allowed to propose a molecular biomarker pattern for this fungus. A. niger is a ubiquitous fungus responsible for food contamination, being reported as one of the main agents of the black mold disease, a serious post-harvest pathology of table grapes. In this work the metabolite pattern already proposed was tested in three different conditions, i.e., by using : (i) 1 day of growth time for A. niger cultures; (ii) 1 day of growth time through A. niger cultures obtained from previously contaminated grapes and iii) in situ SPME approach directly on previously contaminated table grapes with A. niger. Supervised multivariate analysis was performed which revealed that after 1 day of inoculation it was possible to detect the A. niger biomarkers, which allows to infer its presence. Furthermore, the follow-up of this set of metabolites showed that they can be employed to confirm the presence of the pathogen in two varieties of table grapes. The results obtained confirm the potential applicability of the pattern of A. niger biomarkers to early detect the fungi (after 1 day of contamination) and also may be further explored for access food susceptibility to fungi contamination, based on a direct analysis of food item and taking advantage of the high sensitivity of the GC×GC-ToFMS.

Selección de un método de aprendizaje automático para clasificar patrones biomarcadores de lesiones precancerosas de las cuerdas vocales

The National Survey on Drug, Alcohol and Tobacco, 2016-2017, notes that 15.6 million Mexicans are active smokers and, by 2030, expect the death of 8 million cancers of the larynx or lung. Therefore, the World Health Organization (WHO) recommends detecting precancerous lesions of the larynx. This is possible, as they are characterized by a biomarker pattern manifested by the alteration of the biomechanical interpretation of the vocal cords, regardless of the sex and age of the smoker. The goal of this article is to evaluate three machine learning methods: neural networks, Gaussian networks, and decision tree to determine the method that best solves the problem of detecting patterns of precancerous vocal cord injury biomarkers. It uses the WEKA tool and a knowledge bank, endorsed by NOM-012-SSA3-2012, with 250 patterns, and provided by the Luis Guillermo Ibarra National Institute of Rehabilitation, Ibarra. The performance of the methods was compared by ROC curves and confusion matrices, under the criteria established by ISO-5725. The decision tree the method that best responds to the detection of patterns of biomarkers of precancerous lesions of the vocal cords.

Annexin A2 Expression in the Aerogenous Spread of Pulmonary Invasive Mucinous Adenocarcinoma with Gastric Lineage

Spread through air spaces (STAS) is a unique form of lung cancer progression associated with a worse prognosis. However, the mechanisms underlying STAS and the associated proteins remain unclear. Annexin A2 (ANX A2), which is a membrane-binding protein involved in cell adhesion, is known to promote cancer invasion. In this report, we describe the immunohistochemical analysis of ANX A2 expression in an invasive mucinous adenocarcinoma (IMAC) resected from a 63-year-old man in whom the tumor cells had detached from the alveolar wall and exhibited STAS. At the detachment site, we observed cytoplasmic ANX A2 positivity on the basal side and in the exfoliative gap, as well as reduced collagen IV positivity expression. This biomarker pattern suggested an IMAC with gastric lineage. We hypothesize that ANX A2 is secreted from the basal sides of tumor cells and induces tumor cell detachment by degrading the basement membrane. A further comparison of this case with an IMAC with nongastric lineage suggested the following probabilities: (1) ANX A2 likely contributes to STAS in a manner that is dependent on its subcellular localization. (2) Both the subcellular localization of ANX A2 and the detachment site depend on tumor cell characteristics, including the biomarker immunophenotype.

Inflammatory Bowel Disease Types Differ in Markers of Inflammation, Gut Barrier and in Specific Anti-Bacterial Response

Crohn’s disease (CD), ulcerative colitis (UC) and inflammatory bowel disease (IBD) associated with primary sclerosing cholangitis (PSC-IBD), share three major pathogenetic mechanisms of inflammatory bowel disease (IBD)-gut dysbiosis, gut barrier failure and immune system dysregulation. While clinical differences among them are well known, the underlying mechanisms are less explored. To gain an insight into the IBD pathogenesis and to find a specific biomarker pattern for each of them, we used protein array, ELISA and flow cytometry to analyze serum biomarkers and specific anti-microbial B and T cell responses to the gut commensals. We found that decrease in matrix metalloproteinase (MMP)-9 and increase in MMP-14 are the strongest factors discriminating IBD patients from healthy subjects and that PSC-IBD patients have higher levels of Mannan-binding lectin, tissue inhibitor of metalloproteinases 1 (TIMP-1), CD14 and osteoprotegerin than patients with UC. Moreover, we found that low transforming growth factor-β1 (TGF-β1) is associated with disease relapse and low osteoprotegerin with anti-tumor necrosis factor-alpha (TNF-α) therapy. Patients with CD have significantly decreased antibody and increased T cell response mainly to genera Eubacterium, Faecalibacterium and Bacteroides. These results stress the importance of the gut barrier function and immune response to commensal bacteria and point at the specific differences in pathogenesis of PSC-IBD, UC and CD.

Biomarker pattern of ARIA-E participants in phase 3 randomized clinical trials with bapineuzumab

ObjectiveTo evaluate whether amyloid-related imaging abnormalities with edema/effusion (ARIA-E) observed in bapineuzumab clinical trials was associated with specific biomarker patterns.MethodsBapineuzumab, an anti-β-amyloid monoclonal antibody, was evaluated in patients with mild to moderate Alzheimer disease. Amyloid PET imaging, CSF biomarkers, or volumetric MRI (vMRI) were assessed.ResultsA total of 1,512 participants underwent one or more biomarker assessments; 154 developed incident ARIA-E. No differences were observed at baseline between ARIA-E and non-ARIA-E participants in brain amyloid burden by PET, the majority of vMRI measures, or CSF biomarkers, with the exception of lower baseline CSF Aβ42 in APOE ε4 noncarrier ARIA-E vs non-ARIA-E groups (bapineuzumab non-ARIA-E p = 0.027; placebo non-ARIA-E p = 0.012). At week 71, bapineuzumab-treated participants with ARIA-E vs non-ARIA-E showed greater reduction in brain amyloid PET, greater reductions in CSF phosphorylated tau (p-tau) (all comparisons p < 0.01), and total tau (t-tau) (all comparisons p < 0.025), and greater hippocampal volume reduction and ventricular enlargement (all p < 0.05). Greater reduction in CSF Aβ40 concentrations was observed for ARIA-E versus both non-ARIA-E groups (bapineuzumab/placebo non-ARIA-E p = 0.015/0.049). No group differences were observed at week 71 for changes in whole brain volume or CSF Aβ42.ConclusionsBaseline biomarkers largely do not predict risk for developing ARIA-E. ARIA-E was associated with significant longitudinal changes in several biomarkers, with larger reductions in amyloid PET and CSF p-tau and t-tau concentrations, and paradoxically greater hippocampal volume reduction and ventricular enlargement, suggesting that ARIA-E in bapineuzumab-treated cases may be related to increased Aβ efflux from the brain and affecting downstream pathogenic processes.

Regulation of inflammatory pathways in schizophrenia: A comparative study with bipolar disorder and healthy controls

BackgroundImmune-inflammatory processes have been implicated in schizophrenia (SCH), but their specificity is not clear.Main aimTo identify potential differential intra-/intercellular biochemical pathways controlling immune-inflammatory response and their oxidative-nitrosative impact on SCH patients, compared with bipolar disorder (BD) patients and healthy controls (HC).MethodsCross-sectional, naturalistic study of a cohort of SCH patients (n=123) and their controls [BD (n=102) and HC (n=80)].Statistical analysisANCOVA (or Quade test) controlling for age and gender when comparing the three groups, and controlling for age, gender, length of illness, cigarettes per day, and body mass index (BMI) when comparing SCH and BD.ResultsPro-inflammatory biomarkers: Expression of COX-1 was statistically higher in SCH and BD than HC (P<0.0001; P<0.0001); NFκB and PGE2 were statistically higher in SCH compared with BD (P=0.001; P<0.0001) and HC (P=0.003; P<0.0001); NLRP3 was higher in BD than HC (P=0.005); and CPR showed a gradient among the three groups. Anti-inflammatory biomarkers: BD patients had lower PPARγ and higher 15d-PGJ2 levels than SCH (P=0.005; P=0.008) and HC (P=0.001; P=0.001). Differences between SCH and BD: previous markers of SCH (NFκB and PGE2) and BD (PPARγ and 15d-PGJ2) remained statistically significant and, interestingly, iNOS and COX-2 (pro-inflammatory biomarkers) levels were statistically higher in SCH than BD (P=0.019; P=0.040).ConclusionsThis study suggests a specific immune-inflammatory biomarker pattern for established SCH (NFκB, PGE2, iNOS, and COX-2) that differentiates it from BD and HC. In future, their pharmacological modulation may constitute a promising therapeutic target.