Optimization of a fluorescence polarization immunoassay for rapid quantification of deoxynivalenol in durum wheat-based products.

A fluorescence polarization immunoassay previously described for deoxynivalenol (DON) screening in wheat was optimized for the rapid quantification of DON in durum wheat kernels, semolina, and pasta. A background signal was observed in both spiked and naturally contaminated samples, strictly depending on the testing matrix. After subtracting the background DON level for durum wheat (0.27 microg of DON per g), semolina (0.08 microg of DON per g), and pasta (0.04 microg of DON per g), an accurate quantification of DON was possible at levels greater than 0.10 microg/g for all matrices. Average recoveries from spiked samples (0.25 to 1.75 microg/g) were 98, 102, and 101% for wheat, semolina, and pasta, respectively. Comparative analyses of 35 naturally contaminated durum wheat samples, 22 semolina samples, and 26 pasta samples performed by both the fluorescence polarization method and high-pressure liquid chromatography/immunoaffinity cleanup showed a good correlation (r0.995). The fluorescence polarization method showed better accuracy and precision with respect to the high-pressure liquid chromatography method and is suitable for the rapid and quantitative determination of DON in durum wheat-based products at levels foreseen by existing or coming international regulations

Sostanze indesiderate negli alimenti zootecnici: effetti sugli ovini e riflessi nel latte e derivati

Negli ovini da latte l’ingestione di alimenti contaminati da sostanza indesiderate rappresenta un rischio sia per animali al pascolo che per quelli confinati. L’assunzione di composti tossici negli ovini da latte comporta effetti negativi a carico dell’animale oltre che costituire un potenziale elemento di rischio per il consumatore finale nel caso in cui il composto tossico permanga nel latte o nelle carni. I metalli pesanti, le diossine e le micotossine rappresentano le sostanze indesiderate negli alimenti per animali di maggiore interesse per la filiera di produzione degli ovini da latte. La presenza di residui di contaminanti nel latte ovino e nei derivati caseari costituisce un importante indicatore della igienicità della filiera produttiva e allo stesso tempo è ritenuto un marcatore indiretto della salubrità dell’ambiente di produzione. Questo lavoro riassume gli aspetti di maggiore rilievo circa gli effetti dei metalli pesanti, delle diossine e di alcune micotossine sugli ovini, con particolare riferimento al loro trasferimento nel latte e nei derivati

Candida-associated denture stomatitis

Candida albicans is a dimorphic yeast strongly gram positive able to live as normal commensal organism in the oral cavity of healthy people. It is the yeast more frequently isolated in the oral cavity. Under local and systemic factors related to the host conditions, it becomes virulent and responsible of oral diseases known as oral candidiasis. It has been shown that the presence of denture is a predisposing factor to the onset of pathologies related to C. albicans. Clinical studies have shown that C. albicans is not only able to adhere to the mucous surfaces, but also to stick to the acrylic resins of the dental prostheses. Both the plaque accumulated on the denture and the poor oral hygiene contribute to the virulence of Candida, offering the clinical picture of Candida-associated denture stomatitis. The therapeutic strategies currently adopted in the clinical practice to overcome these fungal infections provide for the use of topical and/or systemic antifungal and topical antiseptics and disinfectants, the irradiation with microwaves and the accurate mechanical removal of the bacterial plaque from the denture surfaces and from the underlying mucosa. A correct oral hygiene is important for the control of the bacterial biofilm present on the denture and on the oral mucosa and it is the fundamental base for the prophylaxis and the therapy of the Candidaassociated denture stomatitis

Toxigenic fungi and mycotoxins in a climate change scenario: Ecology, genomics, distribution, prediction and prevention of the risk

Toxigenic fungi and mycotoxins are very common in food crops, with noticeable differences in their host specificity in terms of pathogenicity and toxin contamination. In addition, such crops may be infected with mixtures of mycotoxigenic fungi, resulting in multi-mycotoxin contamination. Climate represents the key factor in driving the fungal community structure and mycotoxin contamination levels pre- and post-harvest. Thus, there is significant interest in understanding the impact of interacting climate change-related abiotic factors (especially increased temperature, elevated CO2 and extremes in water availability) on the relative risks of mycotoxin contamination and impacts on food safety and security. We have thus examined the available information from the last decade on relative risks of mycotoxin contamination under future climate change scenarios and identified the gaps in knowledge. This has included the available scientific information on the ecology, genomics, distribution of toxigenic fungi and intervention strategies for mycotoxin control worldwide. In addition, some suggestions for prediction and prevention of mycotoxin risks are summarized together with future perspectives and research needs for a better understanding of the impacts of climate change scenarios

An In-Silico Pipeline for Rapid Screening of DNA Aptamers against Mycotoxins: The Case-Study of Fumonisin B1, Aflatoxin B1 and Ochratoxin A

Aptamers are single-stranded oligonucleotides selected by SELEX (Systematic Evolution of Ligands by EXponential Enrichment) able to discriminate target molecules with high affinity and specificity, even in the case of very closely related structures. Aptamers have been produced for several targets including small molecules like mycotoxins; however, the high affinity for their respective target molecules is a critical requirement. In the last decade, the screening through computational methods of aptamers for their affinity against specific targets has greatly increased and is becoming a commonly used procedure due to its convenience and low costs. This paper describes an in-silico approach for rapid screening of ten ssDNA aptamer sequences against fumonisin B1 (FB1, n = 3), aflatoxin B1 (AFB1, n = 2) and ochratoxin A (OTA, n = 5). Theoretical results were compared with those obtained by testing the same aptamers by fluorescent microscale thermophoresis and by magnetic beads assay for their binding affinity (KD) revealing a good agreement

Effect of prothioconazole-based fungicides on Fusarium head blight, grain yield and deoxynivalenol accumulation in wheat under field conditions

The effect of triazole-based treatments on Fusarium head blight (FHB), grain yields and the accumulation of deoxynivalenol (DON) in harvested wheat kernels was evaluated by means of twenty multi-site field experiments performed during five consecutive growing seasons (from 2004‒2005 to 2008‒2009) in Italy. Fungicide treatments were carried out on different cultivars of common wheat (cv. Serio, Blasco, Genio and Savio) and durum wheat (cv. Orobel, Saragolla, San Carlo, Levante, Duilio, Karur and Derrik) after artificial inoculation with a mixture of toxigenic Fusarium graminearum and F. culmorum strains. The application of fungicides containing prothioconazole (Proline® or Prosaro®) at the beginning of anthesis (BBCH 61) resulted in a consistent reduction of FHB disease severity (by between 39 and 93%) and DON levels in wheat kernels (by between 40 and 91%) and increased wheat yields (from 0.4 to 5.6 t ha-1, average 2.2 t ha-1), as compared to the untreated/inoculated control. Fungicides containing tebuconazole (Folicur® SE) and cyproconazole plus prochloraz (Tiptor® Xcell) showed a reduced effectiveness compared with prothioconazole-based treatments. All fungicide treatments were more effective in reducing DON and increasing grain yields of common wheat than durum wheat. Results showed that the application of fungicides containing prothioconazole at the beginning of anthesis provided a strong reduction of FHB disease, allowing both an increase in grain yields and a considerable reduction of DON content in wheat kernels

Transcriptional addiction in cancer cells is mediated by YAP/TAZ through BRD4

Cancer cells rely on dysregulated gene expression. This establishes specific transcriptional addictions that may be therapeutically exploited. Yet, the mechanisms that are ultimately responsible for these addictions are poorly understood. Here, we investigated the transcriptional dependencies of transformed cells to the transcription factors YAP and TAZ. YAP/TAZ physically engage the general coactivator bromodomain-containing protein 4 (BRD4), dictating the genome-wide association of BRD4 to chromatin. YAP/TAZ flag a large set of enhancers with super-enhancer-like functional properties. YAP/TAZ-bound enhancers mediate the recruitment of BRD4 and RNA polymerase II at YAP/TAZ-regulated promoters, boosting the expression of a host of growth-regulating genes. Treatment with small-molecule inhibitors of BRD4 blunts YAP/TAZ pro-tumorigenic activity in several cell or tissue contexts, causes the regression of pre-established, YAP/TAZ-addicted neoplastic lesions and reverts drug resistance. This work sheds light on essential mediators, mechanisms and genome-wide regulatory elements that are responsible for transcriptional addiction in cancer and lays the groundwork for a rational use of BET inhibitors according to YAP/TAZ biology

Convergent genetic and expression data implicate immunity in Alzheimer's disease

Background Late–onset Alzheimer's disease (AD) is heritable with 20 genes showing genome wide association in the International Genomics of Alzheimer's Project (IGAP). To identify the biology underlying the disease we extended these genetic data in a pathway analysis. Methods The ALIGATOR and GSEA algorithms were used in the IGAP data to identify associated functional pathways and correlated gene expression networks in human brain. Results ALIGATOR identified an excess of curated biological pathways showing enrichment of association. Enriched areas of biology included the immune response (p = 3.27×10-12 after multiple testing correction for pathways), regulation of endocytosis (p = 1.31×10-11), cholesterol transport (p = 2.96 × 10-9) and proteasome-ubiquitin activity (p = 1.34×10-6). Correlated gene expression analysis identified four significant network modules, all related to the immune response (corrected p 0.002 – 0.05). Conclusions The immune response, regulation of endocytosis, cholesterol transport and protein ubiquitination represent prime targets for AD therapeutics

A novel Alzheimer disease locus located near the gene encoding tau protein

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this recordAPOE ε4, the most significant genetic risk factor for Alzheimer disease (AD), may mask effects of other loci. We re-analyzed genome-wide association study (GWAS) data from the International Genomics of Alzheimer's Project (IGAP) Consortium in APOE ε4+ (10 352 cases and 9207 controls) and APOE ε4- (7184 cases and 26 968 controls) subgroups as well as in the total sample testing for interaction between a single-nucleotide polymorphism (SNP) and APOE ε4 status. Suggestive associations (P<1 × 10-4) in stage 1 were evaluated in an independent sample (stage 2) containing 4203 subjects (APOE ε4+: 1250 cases and 536 controls; APOE ε4-: 718 cases and 1699 controls). Among APOE ε4- subjects, novel genome-wide significant (GWS) association was observed with 17 SNPs (all between KANSL1 and LRRC37A on chromosome 17 near MAPT) in a meta-analysis of the stage 1 and stage 2 data sets (best SNP, rs2732703, P=5·8 × 10-9). Conditional analysis revealed that rs2732703 accounted for association signals in the entire 100-kilobase region that includes MAPT. Except for previously identified AD loci showing stronger association in APOE ε4+ subjects (CR1 and CLU) or APOE ε4- subjects (MS4A6A/MS4A4A/MS4A6E), no other SNPs were significantly associated with AD in a specific APOE genotype subgroup. In addition, the finding in the stage 1 sample that AD risk is significantly influenced by the interaction of APOE with rs1595014 in TMEM106B (P=1·6 × 10-7) is noteworthy, because TMEM106B variants have previously been associated with risk of frontotemporal dementia. Expression quantitative trait locus analysis revealed that rs113986870, one of the GWS SNPs near rs2732703, is significantly associated with four KANSL1 probes that target transcription of the first translated exon and an untranslated exon in hippocampus (P≤1.3 × 10-8), frontal cortex (P≤1.3 × 10-9) and temporal cortex (P≤1.2 × 10-11). Rs113986870 is also strongly associated with a MAPT probe that targets transcription of alternatively spliced exon 3 in frontal cortex (P=9.2 × 10-6) and temporal cortex (P=2.6 × 10-6). Our APOE-stratified GWAS is the first to show GWS association for AD with SNPs in the chromosome 17q21.31 region. Replication of this finding in independent samples is needed to verify that SNPs in this region have significantly stronger effects on AD risk in persons lacking APOE ε4 compared with persons carrying this allele, and if this is found to hold, further examination of this region and studies aimed at deciphering the mechanism(s) are warranted

Detection methods for mycotoxins in cereal grains and cereal products

Analytical methods for mycotoxins in cereals and cereal-based products require three major steps, including extraction, clean-up (to eliminate interferences from the extract and concentrate the analyte), and detection/determination of the toxin (by using suitable analytical instruments/technologies). Clean-up is essential for the analysis of mycotoxins at trace levels, and involves the use of solid phase extraction and multifunctional (e.g. MycoSep®) or immunoaffinity columns. Different chromatographic methods are commonly used for quantitative determination of mycotoxins, including gas-chromatography (GC) coupled with electron capture, flame ionization or mass spectrometry (MS) detectors (mainly for type-A trichothecenes), and high-performance liquid chromatography (HPLC) coupled with ultraviolet, diode array, fluorescence or MS detectors. The choice of method depends on the matrix and the mycotoxin to be analyzed. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is spreading rapidly as a promising technique for simultaneous screening, identification and quantitative determination of a large number of mycotoxins. In addition, commercial immunometric assays, such as enzyme-linked immunosorbent assays (ELISA), are frequently used for screening purposes as well. Recently, a variety of emerging methods have been proposed for the analysis of mycotoxins in cereals based on novel technologies, including immunochromatography (i.e. lateral flow devices), fluorescence polarization immunoassays (FPIA), infrared spectroscopy (FT-NIR), molecularly imprinted polymers (MIPs) and optical biosensors