OCHRATOXIN A RESIDUES IN HUNTED WILD BOAR (SUS SCROFA) FROM TUSCANY

Ochratoxin A (OTA) is a secondary toxic metabolite synthesized by Aspergillus or Penicillium species, which can contaminate various crops. The International Agency for Research on Cancer classified OTA as a group 2B possible human carcinogen. OTA is nephrotoxic, mutagenic, teratogenic and immunosuppressive. OTA can also be present in meat of animals where its presence comes as a result of animal feeding with contaminated grain and feed mixtures. The Italian Ministry of Health Circular No 10, dated 9 June 1999, establishes, as a guideline, a maximum value of 1 μg/kg OTA for swine meat and meat products. The significant increase in the wild boar population has resulted in an increased prevalence of wild boar meat, offal and ready-made products in the food industry. The aim of the present study was to determine OTA concentrations in muscle, kidney and liver of wild boar hunted in Tuscany region. A total of twenty wild boars (male n=11 female n=9) were collected in the Province of Pisa from November 2014 to April 2015, animals have been slaughtered and the carcass weight were determined (from a min. of 14.9 kg and a max. of 72.0 kg). Samples of kidney, liver and muscles from each wild boar were collected and analyzed with an enzymatic digestion clean-up and high-pressure liquid chromatography with fluorescence detection method (1). The highest levels of OTA were found in the kidneys of the twenty wild boar analyzed (0.07- 2.01 μg/kg, mean 0.58±0.63 μg/kg). The levels found in the liver ranged between 0.08- 1.93 μg/kg, (mean 0.53±0.60). The lowest concentrations were found in muscle (0.04- 0.77 μg/kg, mean 0.24±0.24). In eight samples of the tissue samples examined in this study (4 kidney and corresponding 4 liver), the levels of OTA were higher than the guideline level (1 μg/kg) established by the Italian Ministry of Health. The present results are in agreement with a previous study conducted in Calabria in wild boars (2). Swine are particularly sensitive to OTA, kidneys showed the highest accumulation of the latter 101 Società Italiana delle Scienze Veterinarie toxin, followed by liver and muscle tissue, finally the lowest accumulation is represented in adipose tissue. The present results showed the same type of accumulation in wild boar. Traditionally in Tuscany, as in other regions, wild boar meats are used to produce niche products, especially coppa and salami. In agreement with the research of Monaci et al. (3), dried wild boar meat may contribute to overall OTA intake by carry-over effects into processed meats. Monitoring the quality of meat destined for transformation is a priority in order to decrease the possibility of toxin carry-over to humans. The present study confirms that contamination of meat products by OTA represents a potential emerging source of OTA for distinct segments of the Italian population, who are significant consumers of locally-produced wild boar specialties. 1. Luci G., Vanni M., Ferruzzi G., Mani D., Intorre L., Meucci V. 2016. MethodsX 3: 171-177. 2. Bozzo G., Ceci E., Bonerba E., Di Pinto A., Tantillo G., De Giglio E. 2012. Toxins (Basel) 4: 1440-1450. 3. Monaci L., Tantillo G., Palmisano F. 2004. Analytical and Bioanalytical Chemistry 378: 1777- 1782

Determination of ochratoxin A in pig tissues using enzymatic digestion coupled with high-performance liquid chromatography with a fluorescence detector

We present a new method for the rapid analysis of ochratoxin A (OTA) in pig tissues (muscle, liver and kidney) using enzymatic digestion (ED) coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). OTA was digested with a 1% pancreatin solution in a phosphate buffer and then cleaned with ethylacetate. After being evaporated to dryness and re-dissolved, the sample was determined using HPLC-FLD. The method was validated taking into account the currently permitted limit of 1 μg/kg OTA in pork meat and derived products in Italy. The recovery was higher than 90%. Intra- and inter-day repeatability expressed as RSD were less than 7%. The LOD and LOQ were 0.001 and 0.002 μg/kg, respectively. Our method is more efficient, easier, and cheaper than conventional clean-up procedures (liquid–liquid extraction)

HPLC DETERMINATION OF OCHRATOXIN A IN PIG TISSUES USING ENZYMATIC DIGESTION COUPLED TO MOLECULARLY IMPRINTED SOLID PHASE PURIFICATION

Ochratoxin A (OTA) is a mycotoxin produced as a secondary metabolite by various Aspergillus and Penicillium species with nephrotoxic, carcinogenic, immunotoxic and teratogenic potential effects (1). OTA has been found in several food commodities, including cereals, coffee, beer, wine and spices. OTA can also be present in food of farm animals as a result of carryover from animal contaminated feed (2); consequently, a permitted level of 1 μg/kg OTA in pig meat or pig-derived products was set in Italy, as in other countries. Conventional methods for the determination of OTA in animal tissues are performed by extraction with chloroform and followed by cleaning up with immunoaffinity columns or liquid-liquid partitioning (3). These procedures need a large amount of organic solvents which are environmentally harmful and hazardous to humans. The aim of the present study was to develop a new enzymatic digestion method coupled with molecularly imprinted solid phase purification (MISPE) for quantitative determination of OTA in pig tissues (muscle, liver, or kidney). Five grams of sample aliquot were homogenized with 5 ml of a phosphate buffer using an Ultra Turrax. A 2.5 g aliquot of the homogenate was transferred into a tube, incubated for 1 hour at 37°C with 10 ml solution of 1% pancreatin in a phosphate buffer, ultrasonicated at 75 Hz, purified with MISPE columns (pre-conditioned with 3 ml acetonitrile and 3 ml water). OTA elution was performed with methanol/acetic acid 98:2 (v/v). Final eluate was evaporated to dryness and redissolved into 1 ml of HPLC mobile phase and injected into HPLC-FLD. The method was validated according to EU criteria for the confirmatory methods for organic residues and contaminants (4). For all analyzed matrices mean recovery was > 89 %, intra and inter-day repeatability expressed as RSD < 5 % and LOD and LOQ were 0.0018 and 0.0054 μg/kg, respectively. The method can be applied as alternative routine procedure to detect OTA presence in pigs meat products. (1) Malir et al. Ochratoxin A: 50 Years of Research. Toxins, 8:191, 2016. (2) Duarte et al. Ochratoxin A in feed and foodproducing animals: an undesirable mycotoxin with health and performance effects. Vet. Microbiol. 154:1-13, 2011. (3) Monaci et al. Determination of ochratoxin A in pig tissues by liquid-liquid extraction and clean-up and highperformance liquid chromatography. Anal Bioanal Chem 378:1777-1782, 2004. (4) EC (2002) Commission Decision 657/2001 of 12 August 2002 Implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. pp. 8-36

Clinical Outcome after Adoptive Infusion of BPX-501 Cells (donor T cells transduced with iC9 suicide gene) in Children with Thalassemia Major (TM) Given Alfa/Beta T-Cell Depleted HLA-Haploidentical Stem Cell Transplantation (HSCT)

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T-REX OU4 HIRES: the high resolution spectrograph for the E-ELT

The goal of this unit was to consolidate the project for the construction of the high resolution spectrometer of the E-ELT (HIRES). The task included the development of scientific cases and tools to predict the instrumental performances. From the technical point of view it included several R&D activities in collaboration with highly specialized Italian companies; it culminated with the detailed design of a highly modular instrument based on well established technologies. From the management point of view it lead to the consolidation of a large international consortium that spans over 12 countries and includes most of the European and ESO-related institutes interested in high resolution spectroscopy. This consortium is led by INAF; its formal creation is awaiting the official call by ESO for the phase-A study for the HIRES instrument of the E-ELT

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Acute lymphoblastic leukemia with hypereosinophilia in a child: Case report and literature review

Hypereosinophilia in children can be primary or secondary. Numerous malignant diseases can cause hypereosinophilia, but it is seldom caused by acute lymphoblastic leukemia (ALL). In the event of protracted hypereosinophilia, it is extremely important to make a correct differential diagnosis. Case presentation: We present the case of an 11-year-old boy of Moroccan origin with ALL with hypereosinophilic onset (eosinophils in peripheral blood, 10,000/µL) in the absence of other signs of neoplastic disease, and compare this case with 61 similar cases in the literature. Following hospital admission, the patient initially presented with headache-caused nocturnal awakenings, evening fever, and cough, and he also lost approximately 7 kg in weight in a month not associated with sweating or itching. We first performed bone marrow aspiration, which showed an increase in eosinophils without cellular morphological abnormalities, and bone marrow immunophenotyping showed that 4.5% of cells had a phenotype compatible with lymphoid blasts. A lumbar puncture was negative. Given the poor marrow involvement, it was necessary to repeat a new bone marrow aspiration two days later, which showed an increase in blasts to 14%. A concomitant bone marrow biopsy showed an infiltration of blasts typical of B-cell ALL equal to 20–30% with associated hypereosinophilia. Cytogenetic analysis showed an hyperdiploid karyotype: 53–55, XY, +X, add(1)(q21q25), +4, +9, +10, +14, +2, +1, +21/46, XY. Conclusions: ALL is one of the possible causes of persistent hypereosinophilia. In patients with ALL and hypereosinophilia, peripheral hypereosinophilia can precede the appearance of blasts. Due to the negative prognosis and the increased risk of complications in these patients, bone marrow aspiration and biopsy are recommended if common causes of secondary hypereosinophilia are excluded. © 2018 by the authors. Licensee MDPI, Basel, Switzerland

Hemidesmus indicus induces apoptosis via proteasome inhibition and generation of reactive oxygen species

Proteasome inhibition represents an important anticancer strategy. Here, we studied the mechanisms at the basis of the pro-apoptotic activity of the standardized decoction of Hemidesmus indicus, a plant evoking a complex anticancer activity, and explored its inhibition of proteasome activity in human leukemia cells. Additionally, we preliminary tested the cytotoxicity of some H. indicus’s phytochemicals on leukemia cells and their intestinal absorption on a human intestinal epithelium model consisting of a monolayer of differentiated Caco2 cells. We observed a potent antileukemic effect for H. indicus, imputable to the modulation of different critical targets at protein and mRNA levels and the reduction of the 26S proteasome expression. We found that some phytomarkers of H. indicus decoction passed through the enterocyte monolayer. Overall, our study supports the pharmacological potential of H. indicus, which can represent an interesting botanical drug in the oncological area

Determination of ochratoxin A in tissues of wild boar (Sus scrofa) by enzymatic digestion (ED) coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD).

Ochratoxin A (OTA) is a secondary toxic metabolite synthesized by Aspergillus or Penicillium species, which can contaminate various crops. The International Agency for Research on Cancer (IARC) classified OTA as a group 2B possible human carcinogen. The aim of the present study was to assess OTA concentrations in tissues of wild boar (Sus scrofa L.) from Tuscany (Italy). Over a period of 2 years, samples of muscle, liver, and kidney from 48 wild boars were collected and concentrations of OTA were determined by enzymatic digestion (ED) coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). The highest concentrations of OTA were found in the kidneys of the 48 wild boars analyzed. No difference in concentrations was found based on years of collection and sex while a significantly higher OTA concentration was found in the kidney of the young wild boars with respect to the adult one. Monitoring the quality of meat destined for transformation is a priority in order to decrease the possibility of toxin carry-over to humans. The present study showed that contamination of wild boar meat products by OTA represents a potential emerging source of OTA