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Highly purified human menopausal gonadotropin (HP-hMG [Menopur®, Ferring Pharmaceuticals, Saint-Prex, Switzerland]) contains a 1:1 ratio of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). This analysis aimed to assess gonadotropin (FSH, LH and hCG) abundance in HP-hMG and clarify the source of hCG by assessing the presence of sulfated glycans, which are diagnostic for pituitary hCG forms due to their distinct glycosylation patterns. Additionally, the purity of each sample, their specific components, and their oxidation levels were assessed. HP-hMG samples (three of Menopur® and two of Menogon® Ferring Pharmaceuticals, Saint-Prex, Switzerland) were included in the current analyses. Brevactid® (urinary hCG; Ferring Pharmaceuticals, Saint-Prex, Switzerland) and Ovidrel® (recombinant hCG; Merck KGaA, Darmstadt, Germany) were used as control samples. Glycopeptide mapping and analysis of impurities were carried out by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Oxidation was assessed through reducing peptide mapping using LC-MS/MS. The FSH and LH in the HP-hMG samples showed sulfated glycans, while no signals of sulfated glycopeptides were detected on any site of the beta subunit of hCG. HP-hMG test samples presented the same hCG glycan distribution as the control sample (placental hCG, Brevactid®) extracted from the urine of pregnant women, suggesting a non-pituitary source of hCG. Protein impurities were estimated to constitute approximately 20–30% of the entire HP-hMG protein content in the test samples. More than 200 non-gonadotropin proteins were identified in the HP-hMG test samples, of which several were involved in embryonic development or pregnancy. The alpha subunit of the tested samples was strongly oxidized, with a relative abundance of 20% of the total gonadotropin content. Without taking into account all the protein impurities, the beta subunit of LH was detected only in traces (0.9–1.2%) in all tested HP-HMG samples, confirming the data obtained by intact molecule analysis, while high levels of beta hCG (18–47%) were observed. Advanced molecular analysis of HP-hMG indicates a primarily placental origin of hCG, as evidenced by the absence of hCG sulfated glycans and the predominance of placental non-sulfated hCG in LH activity. The analysis revealed 20–30% of protein impurities and a significant presence of oxidized forms in the HP-hMG samples. These findings are critical for understanding the quality, safety, and clinical profile of HP-hMG. [ABSTRACT FROM AUTHOR]

Clinical expression of coronavirus disease 2019 (COVID‐19) infectionis widely variable including fatal cases and patients with mild symptoms and a rapid resolution. We studied saliva from 63 hospitalized COVID‐19 patients and from 30 healthy controls by integrating large‐scale proteomics, peptidomics and targeted metabolomics to assess the biochemical alterations following the infection and to obtain a set of putative biomarkers useful for noninvasive diagnosis. We used an untargeted approach by using liquid chromatography–tandem mass spectrometry (LC–MS/MS) for proteomics and peptidomics analysis and targeted LC–multiple reaction monitoring/MS for the analysis of amino acids. The levels of 77 proteins were significantly different in COVID‐19 patients. Among these, seven proteins were found only in saliva from patients with COVID‐19, four were up‐regulated and three were down‐regulated at least five‐folds in saliva from COVID‐19 patients in comparison to controls. The analysis of proteins revealed a complex balance between pro‐inflammatory and anti‐inflammatory proteins and a reduced amount of several proteins with immune activity that possibly favours the spreading of the virus. Such reduction could be related to the enhanced activity of endopeptidases induced by the infection that in turn caused an altered balance of free peptides. In fact, on a total of 28 peptides, 22 (80%) were differently expressed in severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and control subjects. The multivariate analysis of such peptides permits to obtain a diagnostic algorithm that discriminate the two populations with a high diagnostic efficiency. Among amino acids, only threonine resulted significantly different between COVID‐19 patients and controls, while alanine levels were significantly different between COVID‐19 patients with different severity. In conclusion, the present study defined a set of molecules to be detected with a quick and easy method based on mass spectrometry tandem useful to reveal biochemical alterations involved in the pathogenesis of such a complex disease. Data are available via ProteomeXchange with identifier PXD045612. [ABSTRACT FROM AUTHOR]

Limited or absent activity of the enzyme α-galactosidase A (α-Gal A), due to mutation in the related gene on the X chromosome, leads to the development of a rare hereditary and genetic disease known as Fabry disease (FD). This pathology involves a progressive accumulation in various organs of the substrates of the enzyme e.g., globotriaosylceramide (Gb3) and its deacylated form, globotriaosylsphingosine (Lyso-Gb3), suggesting these molecules as biomarkers of Fabry disease. The present paper describes the development of an analytical strategy for the identification and quantification of Gb3 and Lyso-Gb3, in serum and blood samples by using liquid chromatography (LC) coupled to mass spectrometry in multiple reaction monitoring (MRM/MS) ion mode. The best experimental conditions were obtained by extracting the glycolipids with chloroform/methanol/H2O (2/1/0.3) and by separating them on a C4 column with a linear gradient (A: H2O with 2 mM ammonium formate. B: methanol with 1 mM ammonium formate, both acidified with 0.2% formic acid). The best transitions (a combination of precursor and fragment ions—m/z) were 786.8 m/z > 268.3 m/z for Lyso-GB3, 1137.3 m/z > 264.3 m/z for Gb3, 1039.3 m/z > 264.4 m/z for N-heptadecanoyl-ceramide trihexoside, and 843.5 m/z > 264.3 m/z for N-glycinated lyso-ceramide trihexoside, the latter being used as an internal standard. The developed method provided a reliable, fast, and effective procedure for direct measurements of GB3 and Lyso-GB3 in serum and blood for diagnosis of Fabry disease, suggesting this method as a complementary assay to the current enzymatic test. Therefore, this approach could open new insights into the clinical diagnostics of lysosomal storage disorders. [ABSTRACT FROM AUTHOR]

Removing ink stains from the pages of ancient and valuable books is a very challenging and sometimes impossible task. Only in some circumstances, restorers are able to improve the appearance of an ink-stained page by lightening the spots using an ink eradicator. This paper reports on an approach for the removal of ink stains from a 17th century book using a combination of two commercial houseware products, the rust remover for textile and bleach. The products were chosen on the basis of their active ingredients such as hydrofluoric acid (HF), which facilitates the removal of metal ions contained in the inks (such as iron and chromium) and sodium hypochlorite (NaClO), known for its whitening abilities. The products were first tested by direct application to one of the several stained pages as a control experiment. The ink stain was successfully removed from the book, but by SEM analysis the page results in a clear detachment of the cellulose fibers from the paper pulp, which reflect a significant change in the paper fine structure. Moreover, the mixing of chemicals occurring during this procedure generated toxic vapors and the entire protocol could only be performed under a chemical fume hood. A new procedure was developed to mitigate these negative effects. Each of the two commercial products was dispersed in a 2% agarose solution shortly before its gelification and the two gels thus obtained were applied consecutively on the surface of the paper to be cleaned. The results showed an effective removal of the ink stain from treated page, avoiding irreparable damage to the paper structure and improving the working conditions of the restorers. Hence, the proposed gel cleaning procedure offers three main advantages: i) a simple and practical tool for effective removal of metallic ink stains from the paper surface; ii) reduction of damage caused by aggressive reagents (HF and NaClO) contained in cleaning products for a better conservation of paper fibers; iii) a great decrease of the risk of accidentally exposing the operators to harmful toxic vapor (produced by accidental mixing of aforementioned chemical reagents). [ABSTRACT FROM AUTHOR]

The contamination of environmental sites due to the presence of persistent species represents an important issue to be tackled. In particular, the presence of high levels of metals in soil and surface water is more frequent. One of the metals that sometimes exceeds the permissible limit set by regulatory authorities is copper. For instance, copper-based fungicides are widely used in viticulture. However, copper ions remain in soil and can enter the food chain, posing threats to human health and environmental safety. Although the rapid detection of copper ions using portable sensors is effective in enhancing early warning, it sometimes solves only half of the problem as remediation is not considered. In this paper, we present a novel integrated/portable approach that merges the remediation and sensing of metals by proposing a remediate-and-sense concept. In order to realize this concept, alginate beads were coupled with printed electrochemical strips for on-site copper detection. Within the same architecture, alginate beads were used to remove copper ions from the soil, and printed electrochemical strips were used to evaluate the efficacy of remediation at the point of need. The concept was applied towards soil containing copper ions at the parts per billion level; with few alginate beads and in the absence of additional species, copper ions were quantitatively removed from the matrix; and 3D printing allowed us to combine the printed strips and spheres within a unique tool. The architecture was optimized and the results were compared to inductively coupled plasma-mass spectrometry (ICP-MS) measurements with a recovery percentage of 90%–110%. It should be noted that this novel portable approach may be applied to other pollutants, opening new possibilities for integrated remediation and sensing. [ABSTRACT FROM AUTHOR]

Copper is an essential yet toxic metal and its overload causes Wilson disease, a disorder due to mutations in copper transporter ATP7B. To remove excess copper into the bile, ATP7B traffics toward canalicular area of hepatocytes. However, the trafficking mechanisms of ATP7B remain elusive. Here, we show that, in response to elevated copper, ATP7B moves from the Golgi to lysosomes and imports metal into their lumen. ATP7B enables lysosomes to undergo exocytosis through the interaction with p62 subunit of dynactin that allows lysosome translocation toward the canalicular pole of hepatocytes. Activation of lysosomal exocytosis stimulates copper clearance from the hepatocytes and rescues the most frequent Wilson-disease-causing ATP7B mutant to the appropriate functional site. Our findings indicate that lysosomes serve as an important intermediate in ATP7B trafficking, whereas lysosomal exocytosis operates as an integral process in copper excretion and hence can be targeted for therapeutic approaches to combat Wilson disease.

Simple Summary: Trace elements include both essential and non-essential elements, some of which are pollutants, while others occur naturally in the environment. Imbalances among these elements pose health risks. The wild boar serves as a valuable bioindicator, sharing many behavioral traits with the domestic pig, especially with those extensively bred. This study noted minor geographical variations and observed higher levels of specific trace elements in young boars based on age differences. Although sex had no significant impact, distinctions between wild boars and swine highlighted the dietary and environmental influences on element concentrations, underscoring the wild boar's role as a bioindicator of environmental elements. This study aimed to investigate the exposure of wild boars and swine from semi-extensive farms in the same area to essential and non-essential elements, measuring their concentration in liver and muscle. Furthermore, the study explored the influence of factors such as sex, age, and the sampling location on wild boars. Higher liver element concentrations were observed in both wild boars and swine. Geographical comparisons revealed minor differences. Young wild boars showed significantly higher Cu, Se, Cd, and Cr levels, while older subjects exhibited elevated Mn levels, reflecting age-related element absorption variations. No significant sex-based variations were noted. Comparing wild boars to swine, wild boars had more non-essential elements due to their foraging behavior and a larger home range. Conversely, swine exhibited a greater prevalence of essential elements, potentially resulting from dietary supplementation. [ABSTRACT FROM AUTHOR]