Search

The objectives of this study were to compare the serum and seminal plasma pharmacokinetic profiles of florfenicol (FLO) and florfenicol amine (FLA) after the administration of FLO either by IM or SC routes in beef bulls. Four clinically healthy Hereford bulls underwent a comprehensive physical exam, including breeding soundness examination, CBC, and chemistry profile panel. Bulls were healthy and classified satisfactory potential breeders. In one group (n = 2), a single dose of FLO was administered SC in the middle of the neck at a dose of 40 mg/kg of body weight. In the second group (n = 2), a single dose was administered IM in the muscles of the neck at a dose of 20 mg/kg. Concentrations of FLO and FLA in serum and seminal plasma were determined by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Blood and semen samples were collected before the administration of FLO and at 12, 24, 36, 48, 72, 96, 120, 144, and 168 h after injection. The blood was collected from the coccygeal vessels, and semen was collected by electroejaculation. All samples were immediately refrigerated, processed within the first hour after collection, and finally stored at −80 °C. The mean level of total FLO in serum was higher when administered by the SC route (1,415.5 ng/mL) than by the IM route (752.4 ng/mL; P = 0.001). Differences were observed between the percentage of FLA in serum (1.8%; ranging from 1.3 to 2.9) and in seminal plasma (27.5%; ranging from 15.9 to 34.2; P = 0.0001). The mean level (±SD) of FLA was higher in seminal plasma compared to serum (467 ± 466 ng/mL and 18 ± 16 ng/mL, respectively; P = 0.001). The mean level of total FLO in seminal plasma was 1,454.8 ng/mL for the SC route and 1,872.9 ng/mL for the IM route without differences between the two routes (P = 0.51). Differences in the mean level of total FLO between serum and seminal plasma were detected (1,187 ± 2,069 ng/mL and 1,748 ± 1,906 ng/mL, respectively; P = 0.04). From the present investigation, it was concluded that FLO is a suitable antibiotic based on its pharmacokinetic attributes and may be employed for the treatment of bull genital infections when its use is indicated. To study the pharmacokinetics of FLO in seminal plasma, the analysis of FLA should be incorporated. [ABSTRACT FROM AUTHOR]

Abstract: An analytical method based on high-performance liquid chromatography (HPLC) combined with fluorescence detection (FL) has been developed for the simultaneous determination of fumonisin B1 (FB1) and its totally hydrolized metabolite aminopentol-1 (AP1) in pig liver. The sample preparation is based on a single solid phase extraction (SPE). o-Phthalaldehyde (OPA) was used for pre-column derivatization before the programmed reversed-phase analysis on phenylhexyl column. The developed method shows good repeatibility for inter- and intra-day precision as well as adequate linearity of calibration curves (r2 was 0.9855 for FB1 and 0.9831 for AP1). Average recoveries from the matrix were 93.6% for FB1 and 95.3% for AP1. The limit of quantification (LOQ) in swine liver was 75μg/kg for FB1 and 42μg/kg for AP1. [Copyright &y& Elsevier]

Abstract: Organophosphorus pesticides (OPPs) were determined in matrices of animal origin by dual column capillary gas chromatography using nitrogen–phosphorus detection (NPD). This method was tested on cow milk and on liver and muscle of wild boar. The isolation of these pesticides was performed by liquid partition followed by cleanup with solid phase cartridge (SPE C18), after extraction from the matrix. The analytes identification was obtained by comparing the retention times in two columns with different polarity. The quantification of each OPP was obtained using parathion-ethyl as internal standard. The method was developed in a UNI EN ISO 9001:2000 certified laboratory. The recovery, investigated by analyzing samples spiked at 5, 10 and 50ppb, ranged from 59 to 117% in milk, from 60 to 81% in liver and from 68 to 76% in muscle. The limit of quantification (LOQ) and limit of detection (LOD) were, respectively, 5 and 1ppb for each compound and allowed quantifying the residues below the legal limits. [Copyright &y& Elsevier]

Florfenicol is a broad-spectrum antibiotic belonging to the amphenicols class that inhibits protein synthesis by binding to bacteria's ribosomal subunits. This drug is commonly used in veterinary medicine to treat bacterial infectious diseases in cattle, swine, poultry, and fish. The proposed method uses a quick protein precipitation with acetonitrile for the extraction of florfenicol and florfenicol amine in serum and seminal plasma, followed by analysis in UHPLC-MS/MS for their simultaneous quantification. A BEH C18 reversed-phase column was chosen for analyte separation, allowing to obtaining sharp and symmetrical peak shapes in a chromatographic run of just 3.5 min under programmed conditions. Two specific transitions were observed for each analyte, and florfenicol-d3 was used as the internal standard. The approach was fully validated in each matrix over ranges suitable for field concentrations of florfenicol and florfenicol amine, showing good linearity during each day of testing (R2 always >0.99). Excellent accuracy and precision were demonstrated, for both analytes, by calculated bias always within ±15% and CV% always below 15% at all QC levels tested. The satisfactory outcomes obtained during recovery, matrix effect, and process efficiency investigations in serum and seminal plasma confirmed the strength of the method for the quantification of target compounds. To our knowledge, this is the first LC-MS/MS-validated approach for the quantification of florfenicol and florfenicol amine in serum and seminal plasma and was successfully applied for the determination of their concentration-time profiles in bulls. This paves the way to understanding the pharmacokinetics of this antibiotic and its active metabolite in bull's seminal plasma, which will enable the design of more appropriate treatment protocols. [ABSTRACT FROM AUTHOR]

The marine dinoglagellates Prorocentrium micans, Gonyaulax polyedra, Gymnodinium sp., and Alexandrium tamarense, collected from the Adriatic Sea during red-tide blooms, were cultured to investigate the 4-methyl sterol construents. To ascertain a possible influence of cell age on the 4-methyl sterol content, for one strain (Gymnodinium sp.) we investigated the composition of these constuents at exponential and stationary growing phases. The lipid material extracted with acetone from the lyophilized algal samples was fractionated by thin-layer chromatography. The 4-methyl sterols recovered from the layer were converted into the corresponding OTMS derivatives, Nine of 11 constituents identified by gas chromatography and gas chromatography-mass spectrometry; only two minor constituents were characterized by their gas chromatographic parameters. All free methyl sterols identified in the algal samples had been detected previously in various dinoflagellates. The 4-methyl seterol fractions generally contained very few constituents. Except for the Gymno-dinium sp. sample, collected at the exponential growing phase (GyD2 exp) which, contains 4, 24d-dimethylcholestan3-ol as a unique constituent, dinosterol was the major component. Moreover, 4,24-ethylcholestan 3-ol was also an important consituent of both Prorocentrum and Gonyaulax strains, whereas considerable amounts of dinostanol characterized all the Gymnodinium sp. strains, In adition, the latter contained several minor constituents such as 4-methylcholestan-3-ol, 4,24-dimethylcholesta-22-en-3-ol, and 4-methyl-24-ethylcholestan-3-ol, 4-Methyl-24-methylene-cholestan-3-ol was a constituent of the Gymnodinium sp. sample, collected at the stationary growing phase (GyD2 stat) only, whereas 4-methylgorgostanol was identified only in the Alexandrium tamarense Gt4 strain Except for 4-methyl-24-ethylcholesta-8(14)-en-3-ol, all the methyl sterol constituents from our algae show a saturated polynuclear system. The pathways by which side-chain modification occur in dinoflagellate 4-methyl sterols are considered, and a map of the fragmentation pattern of the trimethylsilyl-4-methyl sterols under electronic impact is also reported. [ABSTRACT FROM AUTHOR]

The objectives of this investigation were to evaluate the pharmacokinetic parameters of oxytetracycline long-acting in plasma and seminal plasma after a single administration through either subcutaneous or intramuscular route at 10 mg/kg or 20 mg/kg dose. Four Simmental bulls, healthy and satisfactory potential breeders, were used. The route of administration either subcutaneous or intramuscular did not affect the mean values for 10 mg/kg dose in plasma (1,470 ng/mL vs. 1,330 ng/mL; P = 0.82) or seminal plasma (5,710 ng/mL vs. 5,390 ng/mL; P = 0.88), or for 20 mg/kg dose in plasma (2,540 ng/mL vs. 2,590 ng/mL; P = 0.96) or seminal plasma (25,600 ng/mL vs. 19,400 ng/mL; P = 0.58), respectively. Comparison between the 10 mg/kg and 20 mg/kg doses showed a difference in terms of mean plasma levels (1400 ng/mL vs. 2570 ng/mL; P = 0.07) and mean seminal plasma levels (6,480 ng/mL vs. 26,200 ng/mL; P = 0.004), respectively. After the dose of 10 mg/kg, plasma C max was 2,841 ng/mL at 12 h (T max) with a half-life of 20.1 h; seminal plasma C max was 11,515 ng/mL at 24 h (T max) with a half-life of 23.7 h. After the dose of 20 mg/kg, plasma C max was 5,269 ng/mL at 12 h (T max) with a half-life of 18.1 h; seminal plasma C max was 55,040 ng/mL at 24 h (T max) with a half-life of 15.7 h. Oxytetracycline long-acting may be an appropriate antibiotic, owing to its pharmacokinetic properties, that could be used for treating bulls' genital infections when its usage is indicated. • Oxytetracycline long-acting presents satisfactory pharmacokinetics properties for the treatment of bull genital infections when its use is indicated. • The levels of oytetracycline in seminal plasma were 6 or more times higher than plasma levels. • The mean residency time of oxytetracycline in seminal plasma was longer than in plasma. [ABSTRACT FROM AUTHOR]

The objective of this investigation was to evaluate the pharmacokinetic parameters of tulathromycin in plasma and semen of beef bulls after administering a single sc dose at two different sites in the neck. Four Simmental bulls with excellent temperament received a comprehensive physical exam that included breeding soundness examination. In addition, blood was collected and analyzed for CBC and chemical panel in order to rule out any subclinical liver or kidney disease. All bulls were diagnosed as healthy and satisfactory potential breeders. The mean plasma levels of tulathromycin for the two neck sites of sc administration were not different between posterior aspect of the ear where it attaches to the head (RP; regio parotidea; 77.9 ± 43.3 ng/mL; X ± SD) and to the middle of the neck (RC; regio collis lateralis; 73.7 ± 39.7 ng/mL; P = 0.84). The mean seminal plasma levels of tulathromycin after administration in the RP was 608 ± 374 ng/mL and for RC was 867 ± 599 ng/mL without differences between both sites (P = 0.29). The mean level of tulathromycin in plasma was 75.8 ± 40.2 ng/mL, which was lower than mean seminal plasma levels of 781 ± 482 ng/mL (P = 0.001). The plasma peak tulathromycin concentration (C max) was 160 ± 27 ng/mL at 21 ± 6 h (T max) post-administration. The seminal plasma C max was 1539 ± 44.4 ng/mL at 33.00 ± 18.00 h (T max) post-administration. The C max between plasma and seminal plasma were different (P = 0.008) without any differences in T max between plasma and seminal plasma (P = 0.35). The terminal half-life for plasma tulathromycin (81.4 ± 27.6 h) showed a tendency to be shorter than in seminal plasma (114.7 ± 21.7; P = 0.10). The plasma area under the curve concentration time from the first to the last sample (AUC 0–last) was 15,440 ± 1717 ng/mL/h, which was significatively smaller compared with 171,071 ± 58,556 ng/mL/h for seminal plasma AUC 0–last (P = 0.01). The plasma means residence time from the first to the last sample (MRT 0-last) was 89.3 ± 5.1 h and it was shorter than for seminal plasma of 96.6 ± 5.0 h (P = 0.05). From the present investigation, it was concluded that tulathromycin is a suitable antibiotic based in its pharmacokinetic properties that could be used for treatment of bull genital infections when its application is indicated. ∗ Tulathromycin presents satisfactory pharmacokinetics properties for the treatment of bull genital infections when its use is indicated. ∗ The levels of tulathromycin in seminal plasma were 10 times higher than plasma levels. ∗ The mean residency time of tulathromycin in seminal plasma was longer than in plasma levels. [ABSTRACT FROM AUTHOR]

Honey, a component of the European diet, faces contamination challenges that impact both consumer and bee health. Glyphosate and glufosinate-based herbicides are widely used in both agriculture and domestic settings, posing potential threats to humans and bees. This study addresses the need for robust analytical methods to detect glyphosate, glufosinate, and their metabolites in honey, considering the complexities of the matrices. Advanced techniques, such as Liquid Chromatography-Mass Spectrometry (LC-MS/MS) and Ion Chromatography-High-Resolution Mass Spectrometry (IC-HRMS) were employed for method validation and monitoring across 97 honey samples. The extraction procedure was optimized, and the validation procedure followed EU Regulation 808/2021 and SANTE 11312/2021 guidelines. LC-MS/MS and IC-HRMS demonstrated comparability and high sensitivity, with RSD r and RSD R values falling within the range of 3%–18% and 6%–22%, respectively, for all analytes considered except for AMPA. AMPA showed CV% values > 25% at the concentration of 5 and 10 ng/g in LC–MS/MS. For IC-HRMS RSD r and RSD R values for all analytes fall within the 3%–14% and 4%–14% range, respectively. Glyphosate was quantified in 12% of the samples, with one sample exceeding Maximum Residue Level (MRL). Glufosinate and its metabolites were not detected in any of the samples. • LC-MS/MS and IC-HRMS are core techniques for polar pesticides control in honey. • Analytical conditions were optimized to enhance performance in both approaches. • Both techniques proved to be suitable for the purpose. • Glyphosate was detected in 50% of honey samples, with one exceeding the MRL. • Metabolites of glyphosate and glufosinate were not detected. [ABSTRACT FROM AUTHOR]

Glyphosate and glufosinate are among the most widely used pesticides in agriculture worldwide. Their extensive use leads to the presence of their residues on crops and in the surrounding environment. Beehives, bees, and apiculture products can represent potential sources for the accumulation of these substances and their metabolites, and the consequences for bee health, as well as the level of risk to human health from consuming contaminated food, are still unclear. Furthermore, information on the contamination levels of honey and other beehive products by these compounds remains poorly documented. This study is part of a broader research effort aimed at developing specific analytical methods for monitoring the level of these contaminants in bee products. The methodology employed enabled the acquisition of preliminary information concerning the levels of glyphosate and glufosinate contamination in honey samples obtained from various retailers in Italy to assess compliance with the limits established by Regulation 293/2013. The liquid chromatography tandem mass spectrometry analysis of the 30 honey samples revealed quantifiable levels of glyphosate in eight samples, with contamination ranging from 5.4 to 138.5 ng/g. Notably, one sample of the wildflower type showed residue levels nearly three times the maximum residue limit. Additionally, trace levels of glyphosate contamination were detected in another ten samples. It is noteworthy that glufosinate and its metabolites were not detected in any of the analyzed samples within the established method's detection ranges. [ABSTRACT FROM AUTHOR]

The assessment of histamine levels in fishery products has emerged as a paramount issue in the context of global food safety, given its profound implications for human health and the consequential impact on food quality and trade. Histamine intoxication, stemming from the ingestion of foods containing heightened histamine levels, results from the bacterial decarboxylation of histidine under conditions of improper handling, processing, or storage. This study endeavors to provide a thorough examination of histamine contamination in frozen-thawed tuna (Thunnus albacares) samples, employing an integrated approach that combines near-infrared spectroscopy (NIRS) with advanced machine learning techniques. One hundred and one samples were considered, and a systematic fortification process was applied to obtain samples with 4 histamine concentrations (0; 50; 150; 250 mg/kg); the fortification levels were confirmed by the LC-MS/MS analysis. Subsequently, NIRS spectra were collected and chemometric analyses, including modified partial least squares regression (MPLS) and support vector machine (SVM), were employed for quantitative and qualitative evaluation, respectively. Histamine quantification through MPLS utilizing the full spectrum exhibited good predictive performance in cross-validation and in hold-out validation (R2 CV = 0.88; R2 P = 0.74, respectively), confirming the potential of NIRS for estimating histamine levels in tuna. SVM classification models, both binary (presence/absence) and multiclass (four levels), demonstrated high accuracy (100% and 93%, respectively). The study highlights the effectiveness of NIRS combined with machine learning for rapid and accurate histamine detection in frozen-thawed tuna, offering a non-destructive, environmentally friendly alternative to traditional methods. This approach holds significant promise for food business operators and regulatory authorities, enhancing product safety, quality control, and decision-making processes related to histamine contamination in the seafood industry. • Complete distinction between tuna samples with and without histamine. • A swift and eco-friendly method for ensuring histamine-free frozen-thawed tuna. • Practical and rapid method to support FBO and Competent Authority in fishery inspection. • Histamine quantification is crucial for fisheries trade decisions. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl substances (PFASs) are environmental contaminants that have been shown to exert toxic effects, which are dependent upon concentration, in animals and humans. No specific data on the exposure of preterm infants to PFASs are available. We aimed to quantify the potential exposure of preterm infants to PFASs through human milk (HM), to be compared to the exposure data recently reported for infants by EFSA. The amount of PFASs in ten preterm (PHM) and ten donor HM (DHM) samples was evaluated, and the expected daily intake (EDI) at full enteral feeding was calculated. This EDI was compared to the mean and the 95th centile dietary exposure ranges at the lower bound for infants issued by EFSA. The calculated median EDI for total PFASs was 20.72 ng/kg/day (range 10.72–107.84) for PHM and 17.92 ng/kg/day (range 6.4–28.96) for DHM, which were both higher than mean exposure ranges reported for infants (2.4–12.2 ng/kg/day). The calculated EDI for DHM was far more similar to the 95th centile (4.5–27.9 ng/kg/day) dietary exposure ranges. For PHM samples, higher EDI values were obtained, with 4 out of 10 samples exceeding the upper limit of the 95th centile range. Conclusion: The exposure of preterm infants to PFASs through HM feeding might exceed reference values reported for older and healthier infants. Given the immunological and developmental vulnerability of preterm infants, the risks related to their exposure to PFASs should be further investigated, also focusing on how maternal exposure and subsequent transfer through HM feeding can be reduced. What is Known: • Perfluoroalkyl substances (PFASs) are environmental contaminants that have been shown to exert toxic effects, which are dependent upon concentration, in animals and humans. The EFSA has recently issued reference values for PFASs exposure for different age groups. • Infants might be exposed to PFASs prenatally, as these substances can cross the placenta, and postnatally, through breastfeeding. No specific data about exposure of preterm infants through human milk (HM) feeding are currently available. What is New: • The exposure of preterm infants to PFASs through HM feeding might exceed reference values reported for older and healthier infants. • Given the immunological and developmental vulnerability of preterm infants, the risks related to their exposure to PFASs deserve further investigation. As HM represents the optimal feeding for preterm infants, it will be fundamental to focus on how maternal exposure and subsequent transfer through HM feeding can be reduced. [ABSTRACT FROM AUTHOR]

Perfluoroalkylated substances (PFASs) are a wide cluster of fluorinated molecules largely engaged industrially and commercially for many purposes. Because of the strength of the fluorine-carbon bond, PFASs show a firm tenacity against thermal degradation, hydrolysis, photolysis and biodegradation. On the other hand, such chemical stability gives them persistent environmental pollutant feature. In 2012, EFSA published a scientific report on PFASs in food, mentioning their adverse effects on health. Based on observational studies evidences, EFSA has recommended a tolerable daily intake (TDI) for the two most known PFASs, i.e. PFOS 150 ng/kg b.w./day and PFOA 1500 ng/kg b.w./day. The aim of this study was to monitor, for the first time, the level of contamination of PFASs in chicken eggs laid in Northern Italy. The eggs were collected from different rearing systems, in order to search a correlation between this variable and the contamination of PFASs. In this study four PFASs [perfluoro-nnonanoic acid (PFNA), perfluoro-noctanoic-acid (PFOA), sodium perfluoro-1-hexanesulfonate (PFHxS) and sodium perfluoro-1-octanesulfonate (PFOS)] were analyzed by liquid chromatography-tandem mass spectrometer (LC-MS/MS). 132 eggs were analyzed, split up in 11 groups according to the geographical origin and rearing system. Results accord with literature data available for chicken eggs: almost all the samples show a PFASs contamination level under the limit of quantification (LOQ) of 0.25 ng/mL. No significant difference results from the rearing system, attesting an equal distribution and a concentration of PFASs detectable under the limit of quantification. [ABSTRACT FROM AUTHOR]

In 2002, researchers from Stockholm University discovered the presence of acrylamide (AA) in processed foods. This substance has been classified as "probably carcinogenic to humans" by the International Agency for Research on Cancer. In response to the alarming finding, the European Commission issued recommendations (2004/394/EC, 2010/307/EU, and 2013/647/EU), guiding food business operators, raising awareness, and promoting good manufacturing practices to minimize AA formation. These efforts laid the foundation for the comprehensive measures in Regulation (EU) 2017/2158. The Regulation implemented specific measures during production to reduce the amount of AA in food. This study monitored the AA levels in 15,674 samples from 12 processed food commodities. Potato-based products and coffee were found to be the main sources of AA exposure. The "baby foods" and "soft bread" food categories had the lowest contamination levels. The data were then compared to the information previously published by the European Food Safety Authority to assess the trend over time and the effectiveness of the mitigation measures. The results showed a decrease in AA contamination levels for most food categories, particularly for baby foods. • 15,674 samples from 12 foods commodities were analyzed for their acrylamide levels. • Comparison with EFSA data indicates lower contamination in most food categories. • The results support Regulation (EU) 2017/2158's mitigation measures. • Self-control plan can prevent market entry of foods exceeding benchmark levels. • Median contamination levels in baby foods had 5-fold decrease compared to EFSA's data. [ABSTRACT FROM AUTHOR]

Nasal administration has been proposed as a potential approach for the delivery of drugs to the central nervous system. Ribavirin (RBV), an antiviral drug potentially useful to treat viral infections both in humans and animals, has been previously demonstrated to attain several brain compartments after nasal administration. Here, a powder formulation in the form of agglomerates comprising micronized RBV and spray-dried microparticles containing excipients with potential absorption enhancing properties, i.e. mannitol, chitosan, and α-cyclodextrin, was developed for nasal insufflation. The agglomerates were characterized for particle size, agglomeration yield, and ex vivo RBV permeation across rabbit nasal mucosa as well as delivery from an animal dry powder insufflator device. Interestingly, permeation enhancers such as chitosan and mannitol showed a lower amount of RBV permeating across the excised nasal tissue, whereas α-cyclodextrin proved to outperform the other formulations and to match the highly soluble micronized RBV powder taken as a reference. In vivo nasal administration to rats of the agglomerates containing α-cyclodextrin showed an overall higher accumulation of RBV in all the brain compartments analyzed as compared with the micronized RBV administered as such without excipient microparticles. Hence, powder agglomerates are a valuable approach to obtain a nasal formulation potentially attaining nose-to-brain delivery of drugs with minimal processing of the APIs and improvement of the technological and biopharmaceutical properties of micronized API and excipients, as they combine optimal flow properties for handling and dosing, suitable particle size for nasal deposition, high surface area for drug dissolution, and penetration enhancing properties from excipients such as cyclodextrins. [ABSTRACT FROM AUTHOR]

The 12 amino acid peptide derived from the Arabidopsis soluble secretory protein CLAVATA3 (CLV3) acts at the cell surface in a signalling system that regulates the size of apical meristems. The subcellular pathway involved in releasing the peptide from its precursor is unknown. We show that a CLV3-GFP fusion expressed in transfected tobacco protoplasts or transgenic tobacco plants has very short intracellular half-life that cannot be extended by the secretory traffic inhibitors brefeldin A and wortmannin. The fusion is biologically active, since the incubation medium of protoplasts from CLV3-GFP-expressing tobacco contains the CLV3 peptide and inhibits root growth. The rapid disappearance of intact CLV3-GFP requires the signal peptide and is inhibited by the proteasome inhibitor MG132 or coexpression with a mutated CDC48 that inhibits endoplasmic reticulum-associated protein degradation (ERAD). The synthesis of CLV3-GFP is specifically supported by the endoplasmic reticulum chaperone endoplasmin in an in vivo assay. Our results indicate that processing of CLV3 starts intracellularly in an early compartment of the secretory pathway and that ERAD could play a regulatory or direct role in the active peptide synthesis. [ABSTRACT FROM AUTHOR]

Nasal administration has been proposed as a potential approach for the delivery of drugs to the central nervous system. Ribavirin (RBV), an antiviral drug potentially useful to treat viral infections both in humans and animals, has been previously demonstrated to attain several brain compartments after nasal administration. Here, a powder formulation in the form of agglomerates comprising micronized RBV and spray-dried microparticles containing excipients with potential absorption enhancing properties, i.e. mannitol, chitosan, and α-cyclodextrin, was developed for nasal insufflation. The agglomerates were characterized for particle size, agglomeration yield, and ex vivo RBV permeation across rabbit nasal mucosa as well as delivery from an animal dry powder insufflator device. Interestingly, permeation enhancers such as chitosan and mannitol showed a lower amount of RBV permeating across the excised nasal tissue, whereas α-cyclodextrin proved to outperform the other formulations and to match the highly soluble micronized RBV powder taken as a reference. In vivo nasal administration to rats of the agglomerates containing α-cyclodextrin showed an overall higher accumulation of RBV in all the brain compartments analyzed as compared with the micronized RBV administered as such without excipient microparticles. Hence, powder agglomerates are a valuable approach to obtain a nasal formulation potentially attaining nose-to-brain delivery of drugs with minimal processing of the APIs and improvement of the technological and biopharmaceutical properties of micronized API and excipients, as they combine optimal flow properties for handling and dosing, suitable particle size for nasal deposition, high surface area for drug dissolution, and penetration enhancing properties from excipients such as cyclodextrins. [ABSTRACT FROM AUTHOR]

Vitamin B12 (cobalamin) is a metal complex composed of a central cobalt ion bonded to six ligands. It is essential for major biological functions such as protein, fat and carbohydrate metabolism, the maintenance of the central nervous system, and the formation of red blood cells. Since mammals cannot synthesize cobalamin, dietary intake represents the only natural source for humans. Dairy products can provide significant levels of cobalamin; moreover, the European Food Safety Authority (EFSA) panel has set the recommended intake at 4 µg/day for adults. Vitamin B12 content was determined in milk and several matrices related to the process of transformation of the residual whey from Parmigiano Reggiano cheese-making to obtain ricotta cheese. In addition, vitamin B12 degradation during ricotta cheese shelf-life was studied. The analyses were performed using an ultra performance liquid chromatography-tandem mass spectrometry method. Results show that vitamin B12 amount in ricotta from dairy and experimental cheese-making brings respectively 1/8 to 1/4 of the adequate intake in adults established by EFSA. In addition, shelf-life experiment shows that cobalamine is fairly rapidly degraded in ricotta: light effect seems to be significant, even if the light exposure is short. The use of photoprotective packaging material increases B12 shelf-life in the early stage of storage. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are pollutants of anthropic origin with possible side effects on human health. Diet, and in particular fish and seafood, is considered the major intake pathway for humans. The present study investigated the levels of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) contamination in twenty-five samples of fresh fillet of five widely consumed fish species purchased from large retailers in Italy, to be used for an estimation of the Italian population exposure to these contaminants. PFOS and PFOA were found in all samples, at concentrations up to 1896 (mean=627 ng/kg) and 487 ng/kg (mean=75 ng/kg), respectively, confirming the role of fish as high contributor to human exposure. However, a remarkable inter-species variability was observed, and multiple factors were suggested as potentially responsible for such differences, suggesting that the preferential consumption of certain species could likely increase the intake, and thus the exposure. The exposure estimates for both average and high fish consumers resulted far below the tolerable daily intakes for PFOS and PFOA in all age groups, confirming the outcomes of EFSA's scientific report. In particular, the calculated total dietary exposure for the 95th percentile consumers belonging to the toddler age class, the most exposed group, resulted equal to 9.72 ng/kg body weight (BW)/day for PFOS and 8.39 ng/kg BW/day for PFOA. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl substances (PFASs) are a family of persistent pollutants of anthropic origin which can reach humans mainly through diet, causing potentially dangerous effects on health. Fish and fishery products are a major source of exposure, but intra- and inter-specific contamination can be extremely variable. In the present study a single species monitoring of the presence of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the two main compounds of the family, was performed on 140 farmed and wild caught European sea basses ( Dicentrarchus labrax ) from different places in the Mediterranean area. The results highlight a strong correlation between the level of contamination and the origin of fishes, if wild or farmed: on average, wild caught sea basses (PFOS: 112-12,405 ng/kg, median 1345 ng/kg; PFOA: 9–487 ng/kg, median 28 ng/kg) showed higher levels than intensively farmed sea basses (PFOS: 11–105 ng/kg, median 32 ng/kg; PFOA: 9–51 ng/kg, median 21 ng/kg). Significant differences among the various rearing systems were also observed, with extensively-farmed subjects presenting relatively higher levels of both compounds compared to intensively farmed. Moreover, a certain variability among wild fish caught from different sampling sites was observed, confirming that PFASs contamination, which reached in some cases noticeable concentrations, might be influenced by the geographical origin. [ABSTRACT FROM AUTHOR]

Bisphenol A (BPA) is a man-made compound, mainly used as a monomer to produce polycarbonate (PC), epoxy resins, non-polymer additives to other plastics, which have many food related applications, such as food storage containers, tableware and internal coating of cans, as well as non-food applications such as electronic equipment, construction materials and medical devices. BPA exposure can occur when the residual monomer migrates into packaged food and beverages. Moreover, due to the ubiquitous presence of this compound, the general population can be exposed to environmental sources such as water, air and soil. Many studies have investigated the potential health hazards associated with BPA, which can elicit toxic and cancerogenic effects on humans. According to the European Food Safety Authority opinion, diet is considered to be the main source of exposure, especially canned food; moreover, among non-canned food, meat and fish products have the highest levels of BPA contamination. This review focuses on BPA contamination in seafood, analysing worldwide literature (from January 2010 to October 2015) on BPA contamination of edible parts. The authors try to identify differences between canned and non-canned seafood in literature, and gaps in the state of art. The data evaluated underline that all concentrations for both canned and non-canned seafood were below the specific migration limit set by the European Community Directive for BPA in food. Moreover, the canned seafood is more contaminated than the non-canned one. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl substances (PFAS) are fully fluorinated compounds widely used during the last 60 yr in the production of multiple industrial and consumer applications, such as food packaging, nonstick cookware, cleaning agents, and many more. These emerging contaminants have recently become of concern for human health because of their potential negative effects. The risk of exposure to PFAS for humans is mainly related to diet, and the increasing interest in food safety has led the European Commission to call Member States to monitor these contaminants in food matrices. The purpose of the present work was to perform the first monitoring on the presence of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the 2 main and most widely investigated molecules of this family, in cow milk commercially available in Italy. We used an analytical protocol consisting of liquid-liquid extraction followed by 2 purification steps through solid-phase extraction cartridges and injection on an ultra-performance liquid chromatography-tandem mass spectroscopy system. The analysis of 67 samples of different types of cow milk from Italy demonstrated that contamination by PFOS was often present, although at relatively low concentrations (up to 97 ng/L), whereas PFOA was rarely found. On the basis of these results and data reported in the literature on this matrix, milk does not seem to be a major source of PFAS compared with other food categories such as fish and seafood. However, variability among different types of milk must be taken into account, and surveys of milk-derived products would be helpful to better define the risk for consumers. [ABSTRACT FROM AUTHOR]

Abstract: Three postharvest icing treatments were implemented on cuttlefish specimens (overall number = 243) after they were hauled onboard: T1-ICO, storage under a 15-mm-thick layer of contact crushed freshwater ice; T2-ICA, storage over a 15-mm-thick layer of contact crushed freshwater ice; T3-NOI, storage without any contact ice, but placed over a 15-mm-thick layer of contact crushed freshwater ice once ashore. All storage trials (n = 6) were conducted at 2 ± 2 °C and were terminated on the sixth day of storage because the aim of the study was to evaluate the effect of realistic icing practices on the freshness quality and shelf -life of cuttlefish, as assessed by sensory (a non-destructive Quality Index Method, QIM), physical (dielectric properties through the Fish Freshness Meter, “295 – Torrymeter”) and chemical (the breakdown products of adenosine 5′–triphosphate, or ATP, as well as relevant ratios among them) methods. The early use of contact ice, either above (T1-ICO) or underneath (T2-ICA) single-layered cuttlefish placed in self-draining polystyrene boxes, was able to slow down ATP degradation. Nevertheless, because three visual traits were significantly worsened in the former treatment, the T2-ICA treatment was suggested as the most proper icing procedure for this cephalopod. The Torrymeter readings had no utility either for predicting the cuttlefish actual/remaining shelf-life for each icing treatment, or for discriminating among them. The percentage ratio between hypoxanthine (Hx) and adenosine 5′–monophosphate (AMP) proved to be the chemical index most highly correlated with ice days. A useful, non-destructive, QIM scheme for cuttlefish was successfully tried and tested in the process. [Copyright &y& Elsevier]

Objective--To evaluate the pharmacokinetics and clinical efficacy of budesonide in dogs with inflammatory bowel disease (IBD). Animals--11 dogs (mean ± SD age, 5.7 ± 3.9 years; various breeds and body weights) with moderate or severe IBD. Procedures--Each dog received a controlled-release formulation of budesonide (3 mg/m², PO, q 24 h) for 30 days (first day of administration was day 1). The concentration of budesonide and its metabolite (16-α-hydroxyprednisolone) was measured via liquid chromatography-tandem mass spectrometry in plasma and urine samples obtained on days 1 and 8 of treatment. On those days, plasma samples were obtained before the daily budesonide administration and 0.5, 1, 2, 4, and 7 hours after drug administration, whereas urine samples were obtained after collection of the last blood sample. A clinical evaluation was performed on the dogs before onset of drug administration and on days 20 and 30 after start of drug administration. Results--The highest plasma concentration of budesonide and 16-α-hydroxyprednisolone on day 1 was detected at 1 hour and at 2 hours after drug administration, respectively. After standardization on the basis of specific gravity, the ratio between urinary concentrations of budesonide and 16-cehydroxyprednisolone was 0.006 and 0.012 on days 1 and 8, respectively. The clinical response was adequate in 8 of 11 dogs. Conclusions and Clinical Relevance-Budesonide was rapidly absorbed and metabolized in dogs with IBD. The drug gradually accumulated, and there was an adequate therapeutic response and no adverse effects. [ABSTRACT FROM AUTHOR]

Abstract: Due to their widespread diffusion, perfluoroalkyl substances (PFASs) have been frequently found in the environment and in several animal species. It has been demonstrated that they can easily reach also humans, mainly through diet. Being lactation a major route of elimination of these contaminants, their occurrence in human milk is of particular interest, especially considering that it generally represents the unique food source for newborns. Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the two most important compounds of this family, have been frequently found in human milk at variable concentrations, but still limited data are available. The present study, the first conducted in Italy capable to detect these pollutants at ultra-trace levels by UPLC-MS/MS, confirmed the role of lactation as a relevant source of exposure for breastfed children. The measured concentrations ranged between 15 and 288 ng/L for PFOS and between 24 and 241 ng/L for PFOA. Moreover, mean concentrations and frequencies of both analytes resulted higher in milk samples provided by primiparous women, suggesting that the risk of intake might be higher for first-borns. Finally, comparing these results with previous data, PFOS gradual decrease over time since year 2000 was confirmed. [Copyright &y& Elsevier]

Abstract: Ribavirin has proved to be effective in vitro against several RNA viruses responsible for encephalitis in humans and animals. However, the in vivo efficacy towards the cerebral viral load seems to be limited by the blood–brain barrier. Since the nose-to-brain pathway has been indicated for delivering drugs to the brain, we investigated here the distribution of ribavirin in the central nervous system (CNS) after intranasal administration. We first tested in vitro ribavirin diffusion from an aqueous solution across a biological membrane, using Franz cells and rabbit nasal mucosa. About 35% of ribavirin permeated in 4h across the mucosa, after reaching steady-state flux in less than 30min. In the first in vivo experiment, ribavirin aqueous solution was administered intranasally to Sprague Dawley rats (10mg/kg). Animals were sacrificed at 10, 20 or 30min after administration to collect brain areas (cerebellum, olfactory bulb, cerebral cortex, basal ganglia and hippocampus) and biological fluids (cerebrospinal fluid and plasma). Ribavirin, quantified by LC–MS/MS spectrometry, was detected at each time point in all compartments with the highest concentration in olfactory bulb and decreasing in rostro-caudal direction. Two subsequent in vivo experiments compared the nasal route (ribavirin solution) with the intravenous one and the nasal administration of ribavirin solution with ribavirin powder (10mg/kg). It was found that 20min after administration, ribavirin concentration in olfactory bulb was similar after intravenous or nasal administration of the ribavirin solution, whereas the powder led to significantly higher levels. Ribavirin was also present in deeper compartments, such as basal ganglia and hippocampus. Even if the mechanisms involved in ribavirin nose-to-brain transport are not clear, these results suggest a rapid extracellular diffusive flux from the nasal epithelium to the olfactory bulb and different CNS areas. [Copyright &y& Elsevier]

Abstract: Perfluorinated compounds (PFCs) are man-made chemicals for which endocrine disrupting properties and related possible side effects on human health have been reported, particularly in the case of an exposure during the early stages of development, (notably the perinatal period). Existing analytical methods dedicated to PFCs monitoring in food and/or human fluids are currently based on liquid chromatography coupled to tandem mass spectrometry, and were recently demonstrated to present some limitations in terms of sensitivity and/or specificity. An alternative strategy dedicated to the analysis of fourteen PFCs in human breast milk was proposed, based on an effective sample preparation followed by a liquid chromatography coupled to high resolution mass spectrometry measurement (LC–HRMS). This methodology confirmed the high interest for HRMS after negative ionization for such halogenated substances, and finally permitted to reach detection limits around the pgmL−1 range with an outstanding signal specificity compared to LC–MS/MS. The proposed method was applied to a first set of 30 breast milk samples from French women. The main PFCs detected in all these samples were PFOS and PFOA with respective median values of 74 (range from 24 to 171) and 57 (range from 18 to 102) pgmL−1, respectively. These exposure data appeared in the same range as other reported values for European countries. [Copyright &y& Elsevier]

Abstract: A liquid chromatography–tandem mass spectrometry method for the quantitative determination of fumonisin FB1, FB2 and their respective hydrolysed metabolites HFB1 and HFB2 in pig liver has been developed and validated. The method was based on an easy extraction procedure followed by SPE purification. Chromatographic separation of the analytes was performed on a C18 column using 0.3% of formic acid in water and acetonitrile as elution solvent. The mass spectrometer operated in the positive electrospray ionisation mode (ESI+) using multiple reaction monitoring (MRM). An intralaboratory validation was carried out with fortified samples for determining precision, trueness, specificity, limit of detection (LOD) and limit of quantification (LOQ). The method showed good performance characteristics and proved to be sensitive, selective and reliable. The LOD was 0.05ngg−1 and the LOQ was 10ngg−1 for all the analytes. The developed method has been applied to seven pig liver samples in order to test its applicability to evaluate exposure of food animals to fumonisins. [ABSTRACT FROM AUTHOR]

Abstract: A rapid and specific liquid chromatography tandem mass spectrometry (LC–MS/MS) method for the determination of ribavirin (RBV) in rat brain was developed. Sample preparation required only two centrifuge steps before LC–MS/MS analysis and the chromatographic separation was achieved in isocratic conditions using an Atlantis T3 column with a nearly totally aqueous (95%) mobile phase. The method showed a good linearity over a concentration range of 5–1000ppb and satisfactory results in terms of accuracy. [Copyright &y& Elsevier]

Abstract: The aim of the present work was to develop a sensitive and selective method for identification and quantification of fumonisin B1 (FB1) in bovine milk. FB1 was isolated by immunoaffinity column and was detected using liquid chromatography coupled with tandem mass spectrometry in positive electrospray ionization (ESI+). The LOQ of the method was 0.1μg/kg that was lower than the others reported in the literature. The high coefficient of determination (R 2 >0.99) obtained in the range of 0.1–10.0μg/kg, the good recovery (84%) and relative standard deviation (7%) of the proposed method ensure correct fumonisin detection in milk even at relatively low concentrations. The developed method was applied on different commercial samples in order to test its efficacy. FB1 was found above the LOQ in eight out of 10 samples analysed and the average level of contamination found was 0.26μg/kg. [Copyright &y& Elsevier]

• Extensive investigation of PFASs in Italian eggs from backyard chickens (EBC). • More contamination in EBC than in eggs from commercial laying hens. • EBC contribute to PFAS intake of up to 29% of tolerable weekly intake in children. Per- and poly-fluoroalkyl substances (PFASs) are persistent and bioaccumulative compounds with adverse impacts on the environment and human health. Diet is one of the main sources of exposure to PFASs. Recently, the EFSA established a tolerable weekly intake (TWI) limit (4.4 ng/kg b.w.) for a mixture of the four major PFASs. Eggs and egg products can contribute to this intake, with their contamination possibly dependent on the husbandry system. Monitoring Italian eggs from backyard chickens revealed a relatively uniform PFAS contamination, with perfluoro-1-octanesulfonate being the most abundant. Contamination was detected to be significantly higher in eggs from backyard chickens than in eggs from commercial laying hens, consistent with a previous Italian study. According to the recently set TWI value, the consumption of eggs from backyard chickens could contribute significantly to dietary intake of PFASs (up to 29% of the TWI in children, considering the lower bound approach). [ABSTRACT FROM AUTHOR]

Vascular and traumatic injuries of the central nervous system are recognized as global health priorities. A polypharmacology approach that is able to simultaneously target several injury factors by the combination of agents having synergistic effects appears to be promising. Herein, we designed a polymeric delivery system loaded with two drugs, ibuprofen (Ibu) and thyroid hormone triiodothyronine (T3) to in vitro release the suitable amount of the anti-inflammation and the remyelination drug. As a production method, electrospinning technology was used. First, Ibu-loaded micro (diameter circa 0.95–1.20 µm) and nano (diameter circa 0.70 µm) fibers were produced using poly(l-lactide) PLLA and PLGA with different lactide/glycolide ratios (50:50, 75:25, and 85:15) to select the most suitable polymer and fiber diameter. Based on the in vitro release results and in-house knowledge, PLLA nanofibers (mean diameter = 580 ± 120 nm) loaded with both Ibu and T3 were then successfully produced by a co-axial electrospinning technique. The in vitro release studies demonstrated that the final Ibu/T3 PLLA system extended the release of both drugs for 14 days, providing the target sustained release. Finally, studies in cell cultures (RAW macrophages and neural stem cell-derived oligodendrocyte precursor cells—OPCs) demonstrated the anti-inflammatory and promyelinating efficacy of the dual drug-loaded delivery platform. [ABSTRACT FROM AUTHOR]

Vitamin B12 is a water-soluble molecule composed of a tetrapyrrolic complex with a cobalt atom at its centre. It is an essential regulatory element, synthesized only by bacteria; for this reason it is present only in food of animal origin and the daily requirement for humans is about 1 to 2 μg. Since milk and dairy products provide a significant dietary cobalamin intake, an ultra performance liquid chromatographytandem mass spectrometry method was applied to samples collected at different stages along the process of cheese making in order to evaluate the distribution of this molecule. In particular, samples of milk, rennet, whey, ricotta cheese, curd, mozzarella cheese and caciotta cheese were analysed. Results showed a level of vitamin B12 about 10 times higher in whey and ricotta cheese with respect to the milk they are derived from. These data would confirm the tendency of cobalamine to concentrate in the proteic fractions along the cheese production process. [ABSTRACT FROM AUTHOR]