Determination of Polycyclic Aromatic Hydrocarbons in Tea Infusions Samples by High Performance Liquid Chromatography with Fluorimetric Detection

This study focuses on the contamination of 15 polycyclic aromatic hydrocarbons (PAHs), recommended by the US Environmental Protection Agency, in 10 tea brands distributed in Italy. Analyses were carried out with a procedure based on saponification, liquidliquid extraction, and PAHs determination by high performance liquid chromatography with fluorescence detector. A comparison with ultrasonic extraction in bath water was also reported. Contamination is expressed as the sum of analyzed PAHs and ranged between 347 and 4120 ng/L with a mean value of 1675 ng/L. PAHs with 3-4 rings were dominant with a contribution of 92%, while 7% and 1% were found for PAHs with 5 and 6 rings, respectively. Moreover, data revealed that three samples exceeded the EU 2008 criteria established for drinking water in which the sum of benzo[k]fluoranthene, benzo [b]fluoranthene, benzo [g,h,i]perylene, and indeno [1,2,3-cd]pyrene is considered (<100 ng/L) and two samples exceeded the 10 ng/L level allowed for benzo [a]pyrene

On-line separation and determination of trivalent and hexavalent chromium with a new liquid membrane annular contactor coupled to inductively coupled plasma optical emission spectrometry

We describe a new on-line sensitive and selective procedure for the determination of trivalent and hexavalent chromium in liquid samples by a tailor-made contactor (TMC), specifically a liquid membrane annular TMC, coupled with inductively coupled plasma with optical detection. The TMC was designed and developed to integrate the extraction and stripping phases of the analyte in one module to minimize the membrane solvent’s consumption and maximize the speed of transport through the liquid membrane. Moreover, the particular geometry studied, which consists of two coaxial hollow fibers, allows the TMC to be used for both separating and preconcentrating purposes. Both (−)-N-dodecyl-N-methylephedrinium bromide (30 mM) in dichloroethane and HNO3 (0.75 M) were used as the liquid membrane and receiving solution, re-spectively. The proposed method’s performance was evaluated in terms of the hexavalent chro-mium extraction efficiency and the coefficient of variation percentages; these were higher than 85% and less than 5%, respectively. In addition, the proposed procedure was applied to two real sam-ples: a tap water sample and an eluate from solid urban waste. In both cases, the analytical per-formances were good and comparable to those obtained using synthetic standard solutions

Is Ferroptosis a Key Component of the Process Leading to Multiorgan Damage in COVID-19?

Even though COVID-19 is mostly well-known for affecting respiratory pathology, it can also result in several extrapulmonary manifestations, leading to multiorgan damage. A recent reported case of SARS-CoV-2 myocarditis with cardiogenic shock showed a signature of myocardial and kidney ferroptosis, a novel, iron-dependent programmed cell death. The term ferroptosis was coined in the last decade to describe the form of cell death induced by the small molecule erastin. As a specific inducer of ferroptosis, erastin inhibits cystine-glutamate antiporter system Xc-, blocking transportation into the cytoplasm of cystine, a precursor of glutathione (GSH) in exchange with glutamate and the consequent malfunction of GPX4. Ferroptosis is also promoted by intracellular iron overload and by the iron-dependent accumulation of polyunsaturated fatty acids (PUFA)- derived lipid peroxides. Since depletion of GSH, inactivation of GPX4, altered iron metabolism, and upregulation of PUFA peroxidation by reactive oxygen species are peculiar signs of COVID-19, there is the possibility that SARS-CoV-2 may trigger ferroptosis in the cells of multiple organs, thus contributing to multiorgan damage. Here, we review the molecular mechanisms of ferroptosis and its possible relationship with SARS-CoV-2 infection and multiorgan damage. Finally, we analyze the potential interventions that may combat ferroptosis and, therefore, reduce multiorgan damage

Use of lipase immobilized on celluse support for cleaning aged oil layers

The present study reports (i) the covalent immobilization of lipase on different cellulose supports (cotton buds, make-up remover pads, cellulose powder, cotton and tissues) using sodium periodate as activating agent and (ii) its application on the aged linseed oil removal from canvas. The optimization of experimental conditions such as pH, temperature and reaction time was performed for both, immobilization procedure and the biocatalyst application. Thus optimal conditions of immobilization were pH 7.0, 20 ºC, 0.3 mg lipase loading per mg support and 200 min reaction time, while those for treating canvas surface, having stratified aged linseed oil were pH 6, 40 ºC and 45 min reaction time. The ability of the immobilized lipase to remove aged oil films was confirmed by UV-Vis spectroscopy, high performance liquid chromatography (HPLC) and scanning electron microscopic (SEM) analysis. O presente estudo relata (i) a imobilização covalente da lipase de Candida rugosa sobre diferentes suportes de celulose (cotonetes, algodão para remoção de maquiagem, pó de celulose, algodão e lenços), usando periodato de sódio como agente ativo, e (ii) suas aplicações na remoção de óleo de linhaça envelhecido, de telas. A otimização das condições experimentais como pH, temperatura e tempo de reação foi realizada para o procedimento de imobilização e para a aplicação do biocatalisador. Assim, as condições ótimas de imobilização foram pH 7,0, 20 ºC, 0,3 mg de lipase por mg do suporte e tempo de reação de 200 min, enquanto as condições necessárias para o tratamento da superfície de telas contendo óleo de linhaça envelhecido, foram pH 6, 40 ºC e tempo de reação de 45 min. A habilidade da lipase imobilizada em remover filmes de óleo envelhecido foi confirmada por análises de espectroscopia UV-Vis, cromatografia líquida de alta eficiência (HPLC) e microscopia eletrônica de varredura (SEM

New Insights into the Role of Ferroptosis in Cardiovascular Diseases

Cardiovascular diseases (CVDs) are the principal cause of disease burden and death worldwide. Ferroptosis is a new formof regulated cell deathmainly characterized by altered ironmetabolism, increased polyunsaturated fatty acid peroxidation by reactive oxygen species, depletion of glutathione and inactivation of glutathione peroxidase 4. Recently, a series of studies have indicated that ferroptosis is involved in the death of cardiac and vascular cells and has a key impact on the mechanisms leading to CVDs such as ischemic heart disease, ischemia/reperfusion injury, cardiomyopathies, and heart failure. In this article, we reviewed the molecular mechanism of ferroptosis and the current understanding of the pathophysiological role of ferroptosis in ischemic heart disease and in some cardiomyopathies. Moreover, the comprehension of the machinery governing ferroptosis in vascular cells and cardiomyocytes may provide new insights into preventive and therapeutic strategies in CVDs

High resolution preparation of monocyte-derived macrophages (MDM) protein fractions for clinical proteomics

<p>Abstract</p> <p>Background</p> <p>Macrophages are involved in a number of key physiological processes and complex responses such as inflammatory, immunological, infectious diseases and iron homeostasis. These cells are specialised for iron storage and recycling from senescent erythrocytes so they play a central role in the fine tuning of iron balancing and distribution. The comprehension of the many physiological responses of macrophages implies the study of the related molecular events. To this regard, proteomic analysis, is one of the most powerful tools for the elucidation of the molecular mechanisms, in terms of changes in protein expression levels.</p> <p>Results</p> <p>Our aim was to optimize a protocol for protein fractionation and high resolution mapping using human macrophages for clinical studies. We exploited a fractionation protocol based on the neutral detergent Triton X-114. The 2D maps of the fractions obtained showed high resolution and a good level of purity. Western immunoblotting and mass spectrometry (MS/MS analysis) indicated no fraction cross contamination. On 2D-PAGE mini gels (7 × 8 cm) we could count more than five hundred protein spots, substantially increasing the resolution and the number of detectable proteins for the macrophage proteome. The fractions were also evaluated, with preliminary experiments, using Surface Enhanced Laser Desorption Ionization Time of Flight Mass Spectrometry (SELDI-TOF-MS).</p> <p>Conclusion</p> <p>This relatively simple method allows deep investigation into macrophages proteomics producing discrete and accurate protein fractions, especially membrane-associated and integral proteins. The adapted protocol seems highly suitable for further studies of clinical proteomics, especially for the elucidation of the molecular mechanisms controlling iron homeostasis in normal and disease conditions.</p

Dexamethasone Predisposes Human Erythroblasts Toward Impaired Lipid Metabolism and Renders Their ex vivo Expansion Highly Dependent on Plasma Lipoproteins

Cultures of stem cells from discarded sources supplemented with dexamethasone, a synthetic glucocorticoid receptor agonist, generate cultured red blood cells (cRBCs) in numbers sufficient for transfusion. According to the literature, however, erythroblasts generated with dexamethasone exhibit low enucleation rates giving rise to cRBCs that survive poorly in vivo. The knowledge that the glucocorticoid receptor regulates lipid metabolism and that lipid composition dictates the fragility of the plasma membrane suggests that insufficient lipid bioavailability restrains generation of cRBCs. To test this hypothesis, we first compared the expression profiling of erythroblasts generated with or without dexamethasone. This analysis revealed differences in expression of 55 genes, 6 of which encoding proteins involved in lipid metabolism. These were represented by genes encoding the mitochondrial proteins 3-Hydroxymethyl-3-Methylglutaryl-CoA lyase, upregulated, and 3-Oxoacid CoA-Transferase1 and glycerol-3-phosphate acyltransferase1, both downregulated, and the proteins ATP-binding cassette transporter1 and Hydroxysteroid-17-Beta-Dehydrogenase7, upregulated, and cAMP-dependent protein kinase catalytic subunit beta, downregulated. This profiling predicts that dexamethasone, possibly by interfering with mitochondrial functions, impairs the intrinsic lipid metabolism making the synthesis of the plasma membrane of erythroid cells depend on lipid-uptake from external sources. Optical and electron microscopy analyses confirmed that the mitochondria of erythroblasts generated with dexamethasone are abnormal and that their plasma membranes present pebbles associated with membrane ruptures releasing exosomes and micro-vesicles. These results indicate that the lipid supplements of media currently available are not adequate for cRBCs. To identify better lipid supplements, we determined the number of erythroblasts generated in synthetic media supplemented with either currently used liposomes or with lipoproteins purified from human plasma [the total lipoprotein fraction (TL) or its high (HDL), low (LDL) and very low (VLDL) density lipoprotein components]. Both LDL and VLDL generated numbers of erythroid cells 3-2-fold greater than that observed in controls. These greater numbers were associated with 2–3-fold greater amplification of erythroid cells due both to increased proliferation and to resistance to stress-induced death. In conclusion, dexamethasone impairs lipid metabolism making ex vivo expansion of erythroid cells highly dependent on lipid absorbed from external sources and the use of LDL and VLDL as lipid supplements improves the generation of cRBCs

Life-Threatening Protein-Losing Enteropathy Due To Human Cytomegalovirus Infection Upon Immunochemotherapy

Immunochemotherapy adverse events affecting the gastrointestinal tract usually consist of self-limiting nausea/ vomiting or diarrhoea, while bleeding and perforation are rare. A 42-year-old woman treated with bendamustine/ rituximab for splenic marginal zone lymphoma developed alife-threatening protein-losing enteropathy that was a diagnosis conundrum, ranging from drug-induced, immunemediated, neoplastic and infectious forms. Suspecting opportunistic viral infection but with unremarkable immunohistochemistry and peripheral blood tests, the diagnosis of Human Cytomegalovirus enteritis was made only by means of quantitative real-time polymerase chain reaction carried out on mucosal specimens. Steroid discontinuation and a prolonged course of antiviral therapy allowed the patient to overcome the critical phase and to achieve gradual normalisation of stool frequency, body mass index, laboratory tests lasting one year, while disappearance of mucosal viral load resulted soon evident. Human Cytom-egalovirus end-organ disease localised at the gastrointestinal tract is a serious condition whose prompt diagnosis and treatment prevents poor patients prognosis

Red Blood Cell Morphologic Abnormalities in Patients Hospitalized for COVID-19

Peripheral blood smear is a simple laboratory tool, which remains of invaluable help for diagnosing primary and secondary abnormalities of blood cells despite advances in automated and molecular techniques. Red blood cells (RBCs) abnormalities are known to occur in many viral infections, typically in the form of mild normo-microcytic anemia. While several hematological alterations at automated complete blood count (including neutrophilia, lymphopenia, and increased red cell distribution width—RDW) have been consistently associated with severity of COVID-19, there is scarce information on RBCs morphological abnormalities, mainly as case-reports or small series of patients, which are hardly comparable due to heterogeneity in sampling times and definition of illness severity. We report here a systematic evaluation of RBCs morphology at peripheral blood smear in COVID-19 patients within the first 72 h from hospital admission. One hundred and fifteen patients were included, with detailed collection of other clinical variables and follow-up. A certain degree of abnormalities in RBCs morphology was observed in 75 (65%) patients. Heterogenous alterations were noted, with spiculated cells being the more frequent morphology. The group with &gt;10% RBCs abnormalities had more consistent lymphopenia and thrombocytopenia compared to those without abnormalities or &lt;10% RBCs abnormalities (p &lt; 0.018, and p &lt; 0.021, respectively), thus underpinning a possible association with an overall more sustained immune-inflammatory “stress” hematopoiesis. Follow-up analysis showed a different mortality rate across groups, with the highest rate in those with more frequent RBCs morphological alterations compared to those with &lt;10% or no abnormalities (41.9%, vs. 20.5%, vs. 12.5%, respectively, p = 0.012). Despite the inherent limitations of such simple association, our results point out towards further studies on erythropoiesis alterations in the pathophysiology of COVID-19