Determination of Perfluorooctanoic Acid (PFOA) in the Indoor Dust Matter of the Sicily (Italy) Area: Analysis and Exposure Evaluations.

Perfluorooctanoic acid (PFOA) in environmental matrices is increasingly being studied due to its environmental persistence, global occurrence, bioaccumulation, and associated human health risks. Some indoor environments can significantly impact the health of occupants due to pollutants in indoor air and household dust. To investigate the potential exposure of individuals to PFOA in specific confined environments, this study reports an analytical method and results concerning the determination of PFOA in household dust, used as a passive sampler. To the best of our knowledge, this paper represents one of the first studies concerning PFOA concentrations in indoor dust collected in the south of Italy, within the European region. A total of twenty-three dust samples were collected from two different areas of Sicily (Palermo and Milena), extracted, and analyzed by an UHPLC-QTOF-MS/MS system. Finally, PFOA exposure was estimated using a new index (Indoor PFOA Exposure Index, IPEX) that incorporates the PFOA levels in dust, exposure time, and the correlation between the PFOA in dust and blood. It was then compared across four different exposure groups, revealing that PFOA exposure for people working in chemistry laboratories was evaluated to be ten times higher than the exposure for homemakers.

Similarity demonstrated between isolated charge variants of MB02, a biosimilar of bevacizumab, and Avastin® following extended physicochemical and functional characterization.

The majority of recombinant mAb products contain heterogeneous charge variants, commonly the result of post-translational modifications occurring during cell culture and accumulated during production, formulation and storage. MB02 is a biosimilar mAb to bevacizumab. Similarity data of charge variants for biosimilars against its reference products must be generated to demonstrate consistency in product quality and to ensure efficacy and safety. The goal of this work was to isolate seven charge variants of MB02 and Avastin® by semi-preparative cation exchange chromatography followed by purity test and extended analytical characterization to prove similarity. Although poor purity obtained for minor variants complicated data interpretation, an in-depth insight into the charge variants pattern of MB02 compared to Avastin® was obtained, contributing to a better understanding of modifications associated to microheterogeneity. To our knowledge, this is the first comparative analytical study of individual charge variants of a bevacizumab biosimilar following a head-to head approach and the most comprehensive N-glycosylation assessment of IgG1 charge variants. Although modifications related to N- and C-terminal, N-glycans, size heterogeneity or deamidation were specifically enriched among low abundant charge variants, they did not affect binding affinity to VEGF or FcRn and in vitro potency compared with the main species or unfractionated material.

Similarity demonstrated between isolated charge variants of MB02, a biosimilar of bevacizumab, and Avastin® following extended physicochemical and functional characterization.

The majority of recombinant mAb products contain heterogeneous charge variants, commonly the result of post-translational modifications occurring during cell culture and accumulated during production, formulation and storage. MB02 is a biosimilar mAb to bevacizumab. Similarity data of charge variants for biosimilars against its reference products must be generated to demonstrate consistency in product quality and to ensure efficacy and safety. The goal of this work was to isolate seven charge variants of MB02 and Avastin® by semi-preparative cation exchange chromatography followed by purity test and extended analytical characterization to prove similarity. Although poor purity obtained for minor variants complicated data interpretation, an in-depth insight into the charge variants pattern of MB02 compared to Avastin® was obtained, contributing to a better understanding of modifications associated to microheterogeneity. To our knowledge, this is the first comparative analytical study of individual charge variants of a bevacizumab biosimilar following a head-to head approach and the most comprehensive N-glycosylation assessment of IgG1 charge variants. Although modifications related to N- and C-terminal, N-glycans, size heterogeneity or deamidation were specifically enriched among low abundant charge variants, they did not affect binding affinity to VEGF or FcRn and in vitro potency compared with the main species or unfractionated material.

Combining P and Zn fertilization to enhance yield and grain quality in maize grown on Mediterranean soils.

The main aim of this study was to elucidate the effect of individual and joint fertilization with P and Zn on maize plants grown on typical Mediterranean soils with a limited Zn availability. For this purpose, we examined the effects of P and Zn fertilization individually and in combination on growth, yield and grain protein content in maize grown in pots filled with three different Mediterranean soils (LCV, FER and INM). Phosphorus and Zn translocation to grain was impaired, and aboveground dry matter and yield at harvest reduced by 8-85% (LCV and FER), in plants treated with Zn or P alone relative to unfertilized (control) plants. In contrast, joint fertilization with P and Zn enhanced translocation of these nutrients to grain and significantly increased aboveground dry matter (30% in LCV, 50% in FER and 250% in INM) and grain Zn availability in comparison with control plants. Also, joint application of both nutrients significantly increased grain P (LCV) and Zn (LCV and FER) use efficiency relative P and Zn, respectively, alone. Yield was increased between 31% in LCV and 121% in FER relative to control plants, albeit not significantly. Fertilization with P or Zn significantly influenced the abundance of specific proteins affecting grain quality (viz., storage, lys-rich and cell wall proteins), which were more abundant in mature grains from plants fertilized with Zn alone and, to a lesser extent, P + Zn. Sustainable strategies in agriculture should consider P-Zn interactions in maize grown on soils with a limited availability of Zn, where Zn fertilization is crucial to ensure grain quality.

Optimum Olsen Phosphorus/ZincDTPA ratio for the initial growth of maize in agricultural soils of the Mediterranean region.

BACKGROUND: Zinc (Zn) deficiency in crops is commonly aggravated by high levels of phosphorus (P) in soil. In this work, the initial performance of pot-growing maize in response to the available P and Zn in soils with low available Zn and to the application of P and Zn fertilizers was investigated. RESULTS: The soils (six non-calcareous and 14 calcareous) ranged widely in available P (Olsen P: 5.5-37.9 mg kg-1 ), were poor in available Zn [diethylenetriaminepentaacetic acid-extractable Zn (ZnDTPA ): 0.20-0.84 mg kg-1 ] and had an Olsen P/ZnDTPA ratio of 13 to 111 mg mg-1 . Soil P application generally increased aerial dry matter (ADM) yield; Zn increased ADM yield mostly when applied in combination with P; and the sole application of Zn increased yield only in a soil with a high (28 mg kg-1 ) Olsen P and a low (0.36 mg kg-1 ) ZnDTPA . The increase in ADM yield resulting from optimal application of P and/or Zn to the soil was modest in soils where the Olsen P/ZnDTPA ratio was 30-60 and Olsen P was >14 mg kg-1 . Zinc uptake by the control plants was correlated with the ZnDTPA of the soil. For a certain ZnDTPA value, the level of plant available Zn was higher in non-calcareous than in calcareous soils. CONCLUSION: Soil application of fertilizer P and Zn, in soils with low levels of available Zn, should not only aim at increasing the available P and Zn levels but also balancing them at the appropriate Olsen P/ZnDTPA ratio, which was found to lie in the 30-60 range in the present study. © 2020 Society of Chemical Industry.

Photochemical emission and fixation of NOX gases in soils.

Gaseous nitrogen oxides (NOx), which result from the combustion of fossil fuels, volcanic eruptions, forest fires, and biological reactions in soils, not only affect air quality and the atmospheric concentration of ozone, but also contribute to global warming and acid rain. Soil NOx emissions have been largely ascribed to soil microbiological processes; but there is no proof of abiotic catalytic activity affecting soil NO emissions. We provide evidence of gas exchange in soils involving emissions of NOx by photochemical reactions, and their counterpart fixation through photocatalytic reactions under UV-visible irradiation. The catalytic activity promoting NOx capture as nitrate varied widely amongst different soil types, from low in quartzitic sandy soils to high in iron oxide and TiO2 rich soils. Clay soils with significant amounts of smectite also exhibited high rates of NOx sequestration and fixed amounts of N comparable to that of NO (nitric oxide) losses through biotic reactions. In these soils, a flux of 100 µg NNO m-2 h-1, as usually found in most ecosystems, could be reduced by these photochemical reactions by more than 60%. This mechanism of N fixation provides new insight into the nitrogen cycle and may inspire alternative strategies to reduce NO emissions from soils.

Different protein expression in normal and dysfunctional platelets from uremic patients.

Although many uremic patients show platelet dysfunctionality, there are others with normal platelet functionality and even with thrombotic tendencies. Our aim was to evaluate changes in the expression of proteins in functional and dysfunctional uremic platelets. Using the platelet function analyzer (PFA-100) assay, uremic patients were divided according to their platelet functionality into normal (n=7) and dysfunctional (n=8). There were no significant differences in the number of circulating platelets and hematocrit and hemoglobin levels. Two-dimensional electrophoresis and mass spectrometry were used to determine and identify changes in protein expression. The closure time (CT) in the PFA-100 assay was significantly prolonged in the dysfunctional uremic platelets. In the dysfunctional platelets, actin-interacting protein-1 isotype 1 was down-regulated, while integrin IIb was up-regulated. Glutathione-S-transferase isotypes 1 and 2 and peroxiredoxin VI were up-regulated in the dysfunctional platelets. Pearson analysis showed a negative correlation between the platelet expression of integrin IIb and creatinine clearance. A positive correlation was found between creatinine clearance and glutathione-S-transferase isotype 2. Serum uric acid concentration was positively correlated with CT values and glutathione-S-transferase isotype 1. In conclusion, the analysis of the protein expression in uremic platelets with normal and dysfunctional activity revealed differences which may occur at the megakaryocyte level.

Inhibition of acyl-CoA cholesterol acyltransferase by F12511 (Eflucimibe): could it be a new antiatherosclerotic therapeutic?

Lipid-lowering strategies, particularly with statins, have been extremely useful in the prevention of cardiovascular disease. However, many patients who receive statin monotherapy do not achieve the desired cardiovascular benefits. Accumulation of cholesteryl esters within macrophages constitutes the hallmark of foam cells during atherogenesis. The action of acyl-coenzyme A (CoA): cholesterol acyltransferase (ACAT) leads to formation of cholesterol esters. There are two different ACAT isoforms: ACAT1 and ACAT2. A considerable interest to develop ACAT inhibitors has been emerging. This review has been focused on the current knowledge about a new ACAT inhibitor, F12511 or eflucimibe, and more particularly on its antiatherosclerotic properties.

Effects of coronary prestenting platelet inhibition on coronary poststenting inflammation.

The aim of this study was to analyze the effect of 2 antiplatelet regimens on the inhibition of GP IIb/IIIa-dependent platelet activation and their association with the poststenting inflammatory response. Seventeen patients with acute myocardial infarction were divided into 2 groups: (A) clopidogrel plus tirofiban infusion administered together during inclusion (n = 10); (B) clopidogrel administered at inclusion and followed 2 hours after by tirofiban (n = 7). Blood samples were obtained at inclusion and at 24 and 48 hours after stenting. Before stenting, a greater reduction of GP IIb/IIIa-dependent platelet activation was found in both groups, although it was greater in group A than in group B. This statistical difference was not observed at 24 and 48 hours after the procedure. At 48 hours after stenting, interleukin-6, interleukin-10, soluble intracellular adhesion molecule-1, and soluble CD40 ligand plasma values were not different between experimental groups. By proteomics, different isoforms of the following proteins were identified: alpha 1-antitrypsin (ATT-1), fibrinogen gamma chain, apolipoprotein A-IV, apolipoprotein A-I, vitamin D binding protein, haptoglobin, and serotransferrin. At 48 hours after stenting, only the plasma expression of the ATT-1 isoform 5 was significantly increased in group A compared with group B. In conclusion, a greater inhibition of GP IIb/IIIa-dependent platelet activation before stenting was not correlated with a different inflammatory activity early after stenting.

Modifications by Olmesartan medoxomil treatment of the platelet protein profile of moderate hypertensive patients.

Olmesartan medoxomil is a new angiotensin II receptor blockers (ARB) which exhibits pleiotropic effects that are not fully understood. Our aims were: i) to determine the effect of Olmesartan medoxomil on blood pressure, lipid profile and renal functionality in moderately hypertensive patients with non-controlled blood pressure, ii) to determine if Olmesartan medoxomil may exert anti-inflammatory effects and modify the expression profile of platelet proteins. Thirteen moderate hypertensive patients with non-controlled systolic blood pressure (SBP) and renal function classified as Kidney Disease Outcome Quality Initiative stage 2-3 were included. Patients were treated with Olmesartan medoxomil (20 mg/day) for 6 months. SBP, proteinuria and the plasma levels of cholesterol and low density lipoprotein (LDL)-cholesterol were reduced after the treatment. Olmesartan medoxomil did not modify the circulating plasma levels of a number of proteins associated with inflammation, but reduced the expression level of different platelet proteins including tropomyosin-ß chain isotypes 3 and 4, serotransferrin isotypes 1 to 5, the leukocyte elastase inhibitor and the chloride intracellular channel-protein isotype 1. The expression of the gelsolin precursor isotype 4 was increased in the platelets after the treatment. In summary, Olmesartan medoxomil reduced SBP, total and LDL-cholesterol plasma levels and urinary protein excretion and induced changes in the expression of platelet proteins which may be related to some action of the drug at the megakaryocyte level.

Relationship between vitamin D binding protein and aspirin resistance in coronary ischemic patients: a proteomic study.

Our aim was to analyze the plasma proteome in aspirin (acetylsalicylic acid [ASA])-sensitive and ASA-resistant coronary ischemic patients. Plasma from 19 ASA-sensitive and 19 ASA-resistant patients was analyzed. For the proteomic study, two-dimensional electrophoresis was performed. The expression of one isotype of the fibrinogen gamma chain and three isotypes of haptoglobin was increased in ASA-resistant patients. Three vitamin D binding protein isotypes were increased in ASA-resistant patients. In vitro incubation of vitamin D binding protein (DBP) with blood from healthy volunteers reduced the inhibitory effect of ASA on thromboxane A2 production. DBP may be a new regulator of the inhibitory effect of ASA on platelets.

Direct effect of F12511, a systemic inhibitor of Acyl-CoA cholesterol acyltransferase on bovine aortic endothelial cells.

F12511(S)-2',3',5'-trimethyl-4'-hydroxy-alpha-dodecylthio-alpha-phenylacetanilide (F12511) is a new Acyl-CoA cholesterol acyltransferase (ACAT) inhibitor that not only reduces the plasma cholesterol levels but also has anti-atherosclerotic actions in animals models. The study's aim was to analyze if F12511 may directly modify the ability of tumor necrosis factor--alpha (TNF-alpha)-incubated bovine aortic endothelial cells (BAEC) to express endothelial nitric oxide synthase (eNOS) protein and inflammatory-related proteins such as platelet endothelial cell adhesion molecule (PECAM) and CD40 ligand (CD40L). The addition of increasing concentrations of F12511 (10 to 10 mol/L) failed to modify the level of eNOS protein expressed in control BAEC. TNF-alpha (10 ng/mL) reduced the expression of eNOS protein. In TNF-alpha--incubated BAEC, F12511 protected eNOS expression in a concentration-dependent manner. TNF-alpha stimulated the expression of both CD40L and PECAM in cultured BAEC. F12511 (10 mol/L) failed to modify the expression of CD40L and PECAM in control and TNF-alpha-incubated BAEC. Reverse transcriptase polymerase chain reaction showed a marked expression of the ACAT-2 isoform and absent of expression of the ACAT-1 isoform in BAEC. The presence of ACAT-2 isoform in BAEC was further confirmed by Western blot. F12511 failed to modify the expression of the proinflammatory associated proteins PECAM and CD40L in the endothelium but protected eNOS expression in the endothelial cells exposed to inflammatory conditions.

Proteomic study of plasma from moderate hypercholesterolemic patients.

Proteomics is a technology to detect and identify several proteins and their isoforms in a single sample. We used proteomics to analyze modifications in the protein map of plasma after simvastatin treatment of moderate hypercholesterolemic patients. Plasma from hypercholesterolemic patients (n = 9) was compared before and after 12 weeks of simvastatin treatment (40 mg/day). Patients with similar cardiovascular risk factors were used as controls (CR group). By using two-dimensional electrophoresis and mass spectrometry, we identified the different protein isoforms. The plasma expression of three fibrinogen gamma chain isoforms (FGG) was enhanced, whereas the expression of two isoforms of the fibrinogen beta chain (FGB) was reduced in the hypercholesterolemic patients compared with the CR group. The expression of apolipoprotein A-IV and three haptoglobin isoforms was higher in hypercholesterolemic patients. Simvastatin treatment modified the plasma expression of FGG chain isoform 1, FGB chain isoforms 1 and 2, vitamin D binding protein isoform 3, apo A-IV, and haptoglobin isoform 2. The modification of FGG chain isoform 1 and FGB chain isoforms 1 and 2 was positively correlated with total plasma cholesterol level. Proteomic analysis of plasma may help to know more in depth the molecular mechanism modified by simvastatin treatment.