Determination of regulated and emerging mycotoxins in organic and conventional gluten-free flours by LC-MS/MS

Gluten-free cereal products have grown in popularity in recent years as they are perceived as “healthier” alternatives and can be safely consumed by celiac patients, and people with gluten intolerance or wheat allergies. Molds that produce mycotoxins contaminate cereal crops, posing a threat to global food security. Maximum levels have been set for certain mycotoxins in cereal flours; however, little is known about the levels of emerging mycotoxins in these flours. The aim of this study was to develop an efficient, sensitive, and selective method for the detection of four emerging (beauvericin and enniatins A1, B, and B1) and three regulated (aflatoxin B1, zearalenone, and deoxynivalenol) mycotoxins in gluten-free flours. Ultrasound-assisted matrix solid-phase dispersion was used in the extraction of these mycotoxins from flour samples. The validated method was utilized for the LC-MS/MS analysis of conventional and organic wholegrain oat and rice flours. Six of the seven target mycotoxins were detected in these samples. Multi-mycotoxin contamination was found in all flour types, particularly in conventional wholegrain oat flour. Despite the low detection frequency in rice flour, one sample was found to contain zearalenone at a concentration of 83.2 µg/kg, which was higher than the level set by the European Commission for cereal flours. The emerging mycotoxins had the highest detection frequencies; enniatin B was present in 53% of the samples at a maximum concentration of 56 µg/kg, followed by enniatin B1 and beauvericin, which were detected in 46% of the samples, and at levels reaching 21 µg/kg and 10 µg/kg, respectively. These results highlight the need to improve the current knowledge and regulations on the presence of mycotoxins, particularly emerging ones, in gluten-free flours and cereal-based product

Green tea and pomegranate extract administered during critical moments of the production cycle improves blood antiradical activity and alters cecal microbial ecology of broiler chickens

Phytobiotics are usually tested in feed and throughout the production cycle. However, it could be beneficial to evaluate their effects when administered only during critical moments, such as changes in feeding phases. The aim of the trial was to investigate the effect of a commercial plant extract (PE; IQV-10-P01, InQpharm Animal Health, Kuala Lumpur, Malaysia) on growth performance, blood antiradical activity and cecal microbiome when administered in drinking water to broiler chickens during the post-hatching phase and at each change of diet. In the experiment, 480 1-day-old male broiler chicks were assigned to two groups in a 50-day trial. Broilers received drinking water (C) or drinking water plus PE (T) at a rate of 2 mL/L on days 0 to 4, 10\u201311 and 20\u2013 21. PE did not affect performance and water intake, while total antiradical activity was improved (p < 0.05). A greater abundance of lactic acid bacteria (false discovery rate (FDR) < 0.05) was found in the T group and the result was confirmed at a lower taxonomic level with higher Lactobacillaceae abundance (FDR < 0.05). Our findings suggest that PE administration during critical moments of the production cycle of broiler chickens may exert beneficial effects at a systemic level and on gut microbial ecology

RIP1-HAT1-SirT complex identification and targeting in treatment and prevention of cancer

Purpose: Alteration in cell death is a hallmark of cancer. A functional role regulating survival, apoptosis, and necroptosis has been attributed to RIP1/3 complexes.Experimental Design: We have investigated the role of RIP1 and the effects of MC2494 in cell death induction, using different methods as flow cytometry, transcriptome analysis, immunoprecipitation, enzymatic assays, transfections, mutagenesis, and in vivo studies with different mice models.Results: Here, we show that RIP1 is highly expressed in cancer, and we define a novel RIP1/3-SIRT1/2-HAT1/4 complex. Mass spectrometry identified five acetylations in the kinase and death domain of RIP1. The novel characterized pan-SIRT inhibitor, MC2494, increases RIP1 acetylation at two additional sites in the death domain. Mutagenesis of the acetylated lysine decreases RIP1-dependent cell death, suggesting a role for acetylation of the RIP1 complex in cell death modulation. Accordingly, MC2494 displays tumor-selective potential in vitro, in leukemic blasts ex vivo, and in vivo in both xenograft and allograft cancer models. Mechanistically, MC2494 induces bona fide tumor-restricted acetylated RIP1/caspase-8-mediated apoptosis. Excitingly, MC2494 displays tumor-preventive activity by blocking 7,12-dimethylbenz(α)anthracene-induced mammary gland hyperproliferation in vivoConclusions: These preventive features might prove useful in patients who may benefit from a recurrence-preventive approach with low toxicity during follow-up phases and in cases of established cancer predisposition. Thus, targeting the newly identified RIP1 complex may represent an attractive novel paradigm in cancer treatment and prevention

Aromatic characterization of Moscato Giallo by GC-MS/MS and stable isotopic ratio analysis of the major volatile compounds

Among the Moscato grapes, Moscato Giallo is a winegrape variety characterized by a highcontent of free and glycosylated monoterpenoids, which gives very aromatic wines. Thearomatic bouquet of Moscato Giallo is strongly influenced by the high concentration of linalool, geraniol, linalool oxides, limonene, α-terpineol, citronellol, HO-trienol, HO-diols, 8-Hydroxylinalool, geranic acid and β-myrcene, that give citrus, rose, and peach notes. Except the quali-quantitative analysis, no investigations regarding the isotopic values of the target volatile compounds are documented in literature. Stable isotope ratio analysis represents a modern and powerful tool used by the laboratories responsible for official consumer protection, for the food quality and genuineness assessment. In this study, samples of Moscato Giallo were collected during the harvest season in 2019 from two Italian regions:Trentino – Alto Adige and Veneto, known lands for the cultivation of this aromatic variety. The flavor compounds were extracted from grapes and wines, after alcoholic fermentation of grape juice, and analysed by GC-MS/MS. The results confirmed the presence of typical terpenoids both in free and glycosylated form, responsible for the characteristic aroma of Moscato Giallo variety. The aromatic compounds were also analysed by GC-C\Py-IRMS for a preliminary investigation. The compound-specific isotope ratio analysis allowed to determine the carbon (δ13C) and hydrogen (δ2H) isotopic signatures of the major volatile compounds for the first time

Can screening for genetic markers improve peripheral artery bypass patency?

AbstractObjective: Three genetic mutations have been associated with an increased risk of thromboembolic events: factor V Leiden R506Q, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T (MTHFR) mutations. The aim of this study was to determine the effect of these mutations on patency of peripheral bypass procedures and preoperative and postoperative thromboembolic events. Methods: Two hundred forty-four randomly selected volunteers participating in the Veterans Affairs Cooperative Study #362 were tested for factor V Leiden, prothrombin, or MTHFR mutations with polymerase chain reaction. Patients enrolled in the study were randomized to receive aspirin therapy or aspirin and warfarin therapy after a peripheral bypass procedure. The frequencies of preoperative and postoperative thromboembolic events and primary patency (PP), assisted primary patency (APP), and secondary patency (SP) rates were compared among carriers of the various mutations. Results: Fourteen patients (5.7%) were heterozygous for the factor V Leiden mutation, seven (2.9%) were heterozygous for the prothrombin mutation, and 108 (44.6%) were heterozygous and 15 (6.2%) homozygous for the MTHFR mutation. After surgery, patients homozygous for the MTHFR gene mutation had increased graft thrombosis, compared with patients who were heterozygous (33.3% versus 11.1%; P = .01), and lower PP, APP and SP rates (P < .05). Furthermore, patients heterozygous for the MTHFR mutation had fewer graft thromboses (11.1% versus 24.4%; P = .01), fewer below-knee amputations (0.9% versus 7.6%; P = .02), and higher PP, APP, and SP rates (PP, 79.6%; APP, 88.9%; SP, 90.7%; P < .05) compared with wild-type control subjects (PP, 63%; APP, 75.6%; SP, 76.5%; P < .05). Conclusion: Patients with either factor V Leiden or prothrombin mutations were not at an increased risk for postoperative graft occlusion or thromboembolic events. Patients heterozygous for MTHFR mutation had a lower risk of graft thrombosis and higher graft patency rates compared with both homozygous and wild-type control subjects. Patients homozygous for the MTHFR mutation had lower graft patency rates compared with patients who were heterozygous, and a trend was seen toward lower patency rates compared with wild-type control subjects. Therefore, screening for the MTHFR gene mutation before surgery may identify patients at an increased risk of graft thrombosis. (J Vasc Surg 2002;36:1198-206.

The effects of superoxide dismutase-rich melon pulp concentrate on inflammation, antioxidant status and growth performance of challenged post-weaning piglets

Piglets can often suffer impaired antioxidant status and poor immune response during post-weaning, especially when chronic inflammation takes place, leading to lower growth rates than expected. Oral administration of dietary antioxidant compounds during this period could be a feasible way to balance oxidation processes and increase health and growth performance. The aim of the trial was to study the effects of an antioxidant feed supplement (melon pulp concentrate) that contains high concentration of the antioxidant superoxide dismutase (SOD) on inflammation, antioxidant status and growth performance of lipopolysaccharide (LPS) challenged weaned piglets. In total, 48 weaned piglets were individually allocated to four experimental groups in a 2 72 factorial design for 29 days. Two different dietary treatments were adopted: (a) control (CTR), fed a basal diet, (b) treatment (MPC), fed the basal diet plus 30 g/ton of melon pulp concentrate. On days 19, 21, 23 and 25 half of the animals within CTR and MPC groups were subjected to a challenge with intramuscular injections of an increasing dosage of LPS from Escherichia coli (serotype 0.55:B5) (+) or were injected with an equal amount of PBS solution ( 12). Blood samples were collected at the beginning of the trial and under the challenge period for interleukin 1\u3b2, interleukin 6, tumour necrosis factor \u3b1, haptoglobin, plasma SOD activity, total antioxidant capacity, reactive oxygen species, red blood cells and plasma resistance to haemolysis, and 8-oxo-7, 8-dihydro-2\u2019-deoxyguanosine. Growth performance was evaluated weekly. A positive effect of melon pulp concentrate was evidenced on total antioxidant capacity, half-haemolysis time of red blood cells, average daily gain (ADG) and feed intake, while LPS challenge increased pro-inflammatory cytokines and haptoglobin serum concentrations, with a reduced feed intake and gain : feed (G : F). The obtained results show that oral SOD supplementation with melon pulp concentrate ameliorates the total antioxidant capacity and the half-haemolysis time in red blood cell of post-weaning piglets, with positive results on growing performance

Evaluation of polyphenolic content and antioxidant activity in agri-food wastes and by-products using chemical and physiological extraction

EU-28 produces about 100 Mtonnes/year of food waste, of which 40% is generated from agri-food industries. Agri-food waste (AFW) and by-products may contain components that could be valorised for their bioactivity. However, the bioaccessibility of these products is highly variable and dependent on a wide range of factors, including food matrix characteristics. Thus, the aim of this study was to determine the total phenolic content and the antioxidant capacity of some by-products and AFW using two different extraction methods. Twelve samples of by products (grape marc, Camelina sativa cake, olive pomace and whey) and 12 samples of AFW (fruits and vegetables waste FVW, citrus pulp, strawberry and orange dried) were processed using: methanol extraction (chemical procedure) and in vitro physiological extraction simulating pig gastro-intestinal tract adapted from Regmi et al., 2009. Soy and wheat were included as controls. Afterwards, the polyphenolic content was assessed by Folin\u2013Ciocalteu assay while antioxidant capacity was determined by 2,2-Azino-bis-3-ethylbenzothiazoline-6-sulfonic Acid (ABTS) assay. Results obtained showed that the chemical extracts of by-products and AFW contain different amount of polyphenols; in particular, as expected, the grape marc showed the highest significant (4.5% w/w; p&lt;.05) polyphenolic content compared to all samples considered. Of note, Camilina sativa cake, olive pomace, FVW, orange and strawberry dried showed a polyphenolic content of 1.3, 0.7, 1.3, 1.6 and 1.3% w/w, respectively. The antioxidant capacity of grape marc exhibited a higher (p&lt;.05) value of 573.6 mmol Trolox equivalent (TE)/g compared to the other samples considered. Camilina sativa cake, olive pomace, FVW, orange and strawberry dried showed an antioxidant capacity of 29.2, 11.0, 28.4, 16.3 and 31.0mmol TE/g. The physiological extraction of grape marcs, Camilina sativa cake, olive pomace, FVW, orange, strawberry dried yielded a polyphenolic content of 3.6, 1.8, 3.2, 3.6 and 2.3% w/w, respectively. The antioxidant capacity showed a similar trend to polyphenolic content after physiological extraction. The results obtained in this study indicate that AFW and by-products could be considered a promising bioaccessible source of antioxidants and phenolic compounds with industrial applications for the production of ingredients to functionalize feedstuffs for monogastric animal

Comparative gene expression profiling reveals partially overlapping but distinct genomic actions of different antiestrogens in human breast cancer cells.

Antiestrogens used for breast cancer (BC) treatment differ among each other for the ability to affect estrogen receptor (ER) activity and thereby inhibit hormone-responsive cell functions and viability. We used high-density cDNA microarrays for a comprehensive definition of the gene pathways affected by 17b-estradiol (E2), ICI 182,780 (ICI), 4OH- tamoxifen (Tamoxifen), and raloxifene (RAL) in ER-positive ZR-75.1 cells, a suitable model to investigate estrogen and antiestrogen actions in hormone-responsive BC. The expression of 601 genes was significantly affected by E2 in these cells; in silico analysis reveals that 86 among them include one or more potential ER binding site within or near the promoter and that the binding site signatures for E2F-1, NF-Y, and NRF-1 transcription factors are significantly enriched in the promoters of genes induced by estrogen treatment, while those for CAC-binding protein and LF-A1 in those repressed by the hormone, pointing to novel transcriptional effectors of secondary responses to estrogen in BC cells. Interestingly, expression of 176 E2- regulated mRNAs was unaffected by any of the antiestrogens tested, despite the fact that under the same conditions the transcriptional and cell cycle stimulatory activities of ER were inhibited. On the other hand, of 373 antiestrogen-responsive genes identified here, 52 were unresponsive to estrogen and 25% responded specifically to only one of the compounds tested, revealing non-overlapping and clearly distinguishable effects of the different antiestrogens in BC cells. As some of these differences reflect specificities of the mechanism of action of the antiestrogens tested, we propose to exploit this gene set for characterization of novel hormonal antagonists and selective estrogen receptor modulators (SERMs) and as a tool for testing new associations of antiestrogens, more effective against BC