Polybrominated dibenzo-p-dioxins and furans (PBDD/Fs) in Italian food: Occurrence and dietary exposure

Human exposure to polychlorinated dioxins and furans (PCDD/Fs) through the dietary pathway is widely recognised and regulations in some regions of the world help to limit food contamination. Similar information on the analogous polybrominated dioxins and furans (PBDD/Fs) is scarce, partly due to the higher threshold to analytical access and unavailability of some standard materials. The analytical methodology developed here determined twelve planar PBDD/F congeners using 13Carbon labelled PBDD/F surrogates and high resolution mass spectrometric detection, and was extensively validated prior to the analysis of a range of commonly consumed Italian foods. The methodology also allowed simultaneous determination of PCDD/Fs and polychlorinated biphenyls (PCBs). The results show that PBDD/Fs occurred in different foods over a range of concentrations from <0.001 pg/g to 4.58 pg/g in fish. The dietary exposure (upper bound) of different Italian population groups, resulting from these occurrence levels was estimated using the toxic equivalency (TEQ) approach that is commonly used for dioxin-like contaminants and ranged from 0.17 to 0.42 pg TEQ/kg bodyweight/day (lower bound – 0.01 pg TEQ/kg bodyweight/day) depending on the population subgroup. Although precautionary, upper bound values may provide a more realistic estimate of toxicity as not all congeners and foods were measured. As expected, children were more highly exposed than adults due to lower body weight. These exposure levels were between a quarter and a third of that arising from the sum of PCDD/Fs and PCBs (0.61 to 1.38 pg WHO-TEQ/kg bodyweight/day), but they contribute to the dioxin-like toxicity. If this data is considered in view of the revised tolerable dioxin-like dietary intake published by EFSA in 2018, it is evident that the tolerable weekly intake of 2 pg/kg bodyweight/week would be exceeded by some of the assessed population sub-groups, or all sub-groups if the cumulative intake is considered

COVID-19 Severity in Multiple Sclerosis: Putting Data Into Context

Background and objectives: It is unclear how multiple sclerosis (MS) affects the severity of COVID-19. The aim of this study is to compare COVID-19-related outcomes collected in an Italian cohort of patients with MS with the outcomes expected in the age- and sex-matched Italian population. Methods: Hospitalization, intensive care unit (ICU) admission, and death after COVID-19 diagnosis of 1,362 patients with MS were compared with the age- and sex-matched Italian population in a retrospective observational case-cohort study with population-based control. The observed vs the expected events were compared in the whole MS cohort and in different subgroups (higher risk: Expanded Disability Status Scale [EDSS] score &gt; 3 or at least 1 comorbidity, lower risk: EDSS score ≤ 3 and no comorbidities) by the χ2 test, and the risk excess was quantified by risk ratios (RRs). Results: The risk of severe events was about twice the risk in the age- and sex-matched Italian population: RR = 2.12 for hospitalization (p &lt; 0.001), RR = 2.19 for ICU admission (p &lt; 0.001), and RR = 2.43 for death (p &lt; 0.001). The excess of risk was confined to the higher-risk group (n = 553). In lower-risk patients (n = 809), the rate of events was close to that of the Italian age- and sex-matched population (RR = 1.12 for hospitalization, RR = 1.52 for ICU admission, and RR = 1.19 for death). In the lower-risk group, an increased hospitalization risk was detected in patients on anti-CD20 (RR = 3.03, p = 0.005), whereas a decrease was detected in patients on interferon (0 observed vs 4 expected events, p = 0.04). Discussion: Overall, the MS cohort had a risk of severe events that is twice the risk than the age- and sex-matched Italian population. This excess of risk is mainly explained by the EDSS score and comorbidities, whereas a residual increase of hospitalization risk was observed in patients on anti-CD20 therapies and a decrease in people on interferon

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR&nbsp;=&nbsp;2.05, 95%CI&nbsp;=&nbsp;1.39–3.02, p&nbsp;&lt;&nbsp;0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR&nbsp;=&nbsp;0.42, 95%CI&nbsp;=&nbsp;0.18–0.99, p&nbsp;=&nbsp;0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon

Determination of maize and grain herbicides and their transformation products in soil by use of soil column extraction then liquid chromatography with tandem mass spectrometry

A multiresidue method has been developed for identificationand quantitationof theherbicides most commonly used in cultivationof maize and grain,andof their transformation products, in soil samples. The analytes were isolated by soil column extraction (SCE)and the extracts were purified by useof a Carbograph-1 cartridge. Analysis was performed by liquid chromatography coupled with tandem mass spectrometric detection (LC-TISP-MS-MS) in negative-ion mode. To optimize the extraction conditions affecting the performanceof SCE, e.g. the extracting solvent used, temperature, extracting volume,and solvent flow rate, were Studied. To evaluate the matrix effect in SCE, recovery experiments were performed on soil samples, with different physicaland chemical characteristics, fortified with the 100 ng g(-1)of the target compounds. Recovery data were satisfactoryand the method detection limits were between 3and 100 ng g(-1), depending on the compound

Analytical Methodologies for the Determination and Occurrence of Endocrin Disrupting Chemicals in Sewage Treatment Plants and Natural Waters

In this study, a method for assessing the occurrence of trace amounts of 12 representative estrogenic compounds in sewage and surface waters was developed. The selected substances were the phytoestrogens daidzein, genistein and biochanin A, the alkylphenols bisphenol A and 4-nonylphenol, the natural hormones 17beta-estradiol, estrone, estriol and the synthetic hormone 17alpha-ethynylestradiol and the mycoestrogens zearalenone and two of its metabolites (alpha-zearalanol and beta-zearalanol). The procedure consists in solid phase extraction (SPE) performed with OASIS cartridges followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS-MS). Recoveries were all above 80% for each analyzed aqueous matrices. The developed method was applied to verify the occurrence of endocrine disrupters in environmental samples of sewage influents and effluents of an Italian STP. Phytoestrogens were present in effluents at concentrations ranging from 3 to 83 ng/l, whereas the levels detected for alkylphenols were in the range 13-36 ng/l for bisphenol A and up to 1 mug/l for nonylphenol. Estrogens and zeranols were determined at levels below 30 ng/l. Analysis of a river (Tiber) receiving effluent waters revealed high quantities of bisphenol A (15-29 ng/l) and nonylphenol (up to 1.2 mug/l), whereas the presence of all the other compounds were at levels of few ng/l

Occurrence and determination of herbicides and their major transformation products in environmental waters

A method for the simultaneous determination of selected maize and grain herbicides (amidosulfuron, N-([{[(4,6-dimethoxy-2-pyrimidinyl)amino]carbonyl}amino)sulfonyl]-N-methylmethanesulfonamide; bentazone, 3-(1-methylethyl)-1H-2,1,3-benzothiadiazin-4(3H)-one2,2-dioxide; 2,4-D, (2,4-dichlorophenoxy)acetic acid; MCPA, (4-chloro-2-methylphenoxy)acetic acid; metosulam, N-(2,6'-dichloro-3-methylphenyl)-5,7-dimethoxy[1,2,4]triazolo[1,5-a]pyrimidine-2-sulfonamide) and their transformation products (TPs) (8-hydroxybentazone, 1H-2,1,3-benzothiadiazin-4(3H)-one,8-hydroxy-3-(1-methylethyl)-2, 2-dioxide; bifenox acid, 5-(2,4-dichlorophenoxy)-2-nitrobenzoic acid; clodinafop acid; 2,4-dichlorophenol; 4-chloro-2-methylphenol) in environmental waters was developed. The procedure combines a solid-phase extraction (SPE) with column liquid chromatography-tandem mass spectrometry detection utilizing a pneumatically assisted and heat-assisted electrospray interface in negative mode. The extraction of analytes from filtered drainage or river water samples was performed using as sorbent a graphitized carbon. The recovery, for drinking water, ranged from 91 to 98%, and from 84 to 95% for river water. Excellent method precision was demonstrated with percent relative standard deviation of less than 10% for all analytes at fortification level. Detection limits were at level of few nanograms per liter, except for 2,4-dichlorophenol and 4-chloro-2-methylphenol, that was in the range of tens of nanogram per liter. The present methodology was used for searching in river and drainage waters of maize-grain fields of the Tiber valley, during a typical period of herbicides application from April to July 2001. The levels detected of four compounds were in the range of: non-detectable (ND)-6.1, ND-27, ND-1.9 and ND-0.5 mug l(-1), for MCPA, bentazone, 8-hydroxybentazone and 2,4-D, respectively. Detectable amounts of bentazone-8-hydroxy at level of 0,3 mug l(-1) was found to be transported into river. The highly satisfactory results for all target analytes demonstrate the potential of liquid chromatography-tandem mass spectrometry for the identification and the quantification of the polar and acidic herbicides at below 100 ng l(-1) levels. (C) 2002 Elsevier Science B.V. All rights reserved

Sample preparation for determination of macrocyclic lactone mycotoxins in fish tissue, based on on-line matrix solid-phase dispersion and solid-phase extraction cleanup followed by liquid chromatography/tandem mass spectrometry

A new method based on matrix solid-phase dispersion (MSPD) on-line with a solid-phase extraction (SPE) cleanup process followed by liquid chromatography with tandem mass spectrometry (LC/MS/MS) is presented for the determination of 3 macrocyclic lactone mycotoxins in fish tissues: zearalenone, alpha-zearalenol, and beta-zearalenol. The sample was prepared in a device that used a reversed-phase material (C-18) or a normal-phase material (neutral alumina) as a matrix dispersing agent, and a graphitized carbon black cartridge was used for sequential cleanup by SPE. LC/MS/MS was used for selective determination. Isocratic elution with acetonitrile-methanol-water was used for LC separation; for MS/MS, 2 types of interfaces (a pneumatically assisted electrospray ionization interface or an atmospheric pressure chemical ionization interface) were evaluated and compared in terms of the intensity of the total ion current produced by each analyte. The use of highly selective MSPD on-line with SPE for sample preparation before analysis allowed the removal of interfering matrix compounds present in tissue extracts that would otherwise cause severe ionization suppression of zearalenone and its metabolites during the ionization process. Average recoveries at 100 ng/g were between 83 and 103 % with C-18 and greater than or equal to67% with neutral alumina; the relative standard deviations were <11% with C-18 and <18% with alumina. The limits of detection ranged from 0.1 to 1.0 ng/g. Sample preparation is simple to perform, no special technical equipment is required, and solvent volumes are minimal

Salivary gland transcriptome of Varroa destructor reveals suitable targets for dsRNA-mediated control of the mite

Varroa destructor is a parasitic mite of honey bees and a major driver of honey bee colony losses. By feeding on the host’s body fluids, this obligate ectoparasite has a strong impact on honey bee physiology, causing the reduction of weight and longevity, and the spread of viral pathogens. Despite its importance, many aspects of Varroa lifestyle are still obscure. During feeding activity on the host, the mite injects salivary secretions only partly identified so far. Here we performed a differential expression analysis between the transcriptome of salivary glands and the rest of the body of V. destructor, identifying 12 transcripts encoding putatively secreted proteins. Using databases such as InterPro, UniProt and NCBI, we functionally annotated this secretome, which includes proteins belonging to cysteine protease, serine protease, endochitinase and phospholipase families. Selected genes were subsequently studied from a functional point of view, by analyzing their impact on mite survival following gene silencing. Mites were soaked in dsRNA solutions targeting chitin-binding, kazal serine protease inhibitor and papain cysteine protease, for a few hours and maintained on honey bee pupae under laboratory conditions. The effect on mortality was checked daily, while gene silencing was assessed at 72 hours after treatment, by qRT-PCR. The dsRNA mix targeting the three genes simultaneously resulted effective in reducing mite survival compared to controls. Further functional characterization of these salivary components will offer new knowledge on the molecular basis of Varroa-bee interactions, on which to develop innovative and eco-sustainable mite control strategies