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In recent years, the widespread use of neonicotinoids in agricultural areas has caused environmental pollution due to its lower toxicity to mammals. Honey bees, which are considered as biological indicators of environmental pollution, can carry these pollutants to the hives. Forager bees returning from sunflower crops that have been treated with neonicotinoids treated sunflower fields cause residue accumulation in the hives, which reason colony-level adverse effects. This study analyses neonicotinoid residues in sunflower (Helianthus annuus) honey sampled by beekeepers from Tekirdag province. Honey samples have been subjected to liquid-liquid extraction methods before liquid chromatography-mass spectrometry (LC-MS/MS). The method validation was carried out to fulfill all the necessary requirements of procedures SANCO/12571/2013. Accuracy was in the range of 93.63-108.56%, for recovery in the range of 63.04-103.19%, and for precision in the range 6.03-12.77%. Detection and quantification limits were determined according to the maximum residue limits of each analyte. No neonicotinoid residues were found above the maximum residue limit in the sunflower honey samples analysed. Scientific Research Project Coordination Unit of Tekirdag Namik Kemal University [NKUBAP.00.23, AR.14.05] AcknowledgmentsThis work was supported by the Scientific Research Project Coordination Unit of Tekirdag Namik Kemal University, Tekirdag (project number: NKUBAP.00.23.AR.14.05).

A simple, rapid method using CE and microchip electrophoresis with (CD)-D-4 has been developed for the separation of four nonsteroidal anti-inflammatory drugs (NSAIDs) in the environmental sample. The investigated compounds were ibuprofen (IB), ketoprofen (KET), acetylsalicylic acid (ASA), and diclofenac sodium (DIC). In the present study, we applied for the first time microchip electrophoresis with (CD)-D-4 detection to the separation and detection of ASA, IB, DIC, and KET in the wastewater matrix. Under optimum conditions, the four NSAIDs compounds could be well separated in less than 1 min in a BGE composed of 20 mM His/15 mM Tris, pH 8.6, 2 mM hydroxypropyl-beta-cyclodextrin, and 10% methanol (v/v) at a separation voltage of 1000-1200 V. The proposed method showed excellent repeatability, good sensitivity (LODs ranging between 0.156 and 0.6 mg/L), low cost, high sample throughputs, portable instrumentation for mobile deployment, and extremely lower reagent and sample consumption. The developed method was applied to the analysis of pharmaceuticals in wastewater samples with satisfactory recoveries ranging from 62.5% to 118%.

Subcellular membrane systems are highly enriched in dolichol, whose role in organelle homeostasis and endosomal-lysosomal pathway remains largely unclear besides being involved in protein glycosylation. DHDDS encodes for the catalytic subunit (DHDDS) of the enzyme cis-prenyltransferase (cis-PTase), involved in dolichol biosynthesis and dolichol-dependent protein glycosylation in the endoplasmic reticulum. An autosomal recessive form of retinitis pigmentosa (retinitis pigmentosa 59) has been associated with a recurrent DHDDS variant. Moreover, two recurring de novo substitutions were detected in a few cases presenting with neurodevelopmental disorder, epilepsy and movement disorder. We evaluated a large cohort of patients (n = 25) with de novo pathogenic variants in DHDDS and provided the first systematic description of the clinical features and long-term outcome of this new neurodevelopmental and neurodegenerative disorder. The functional impact of the identified variants was explored by yeast complementation system and enzymatic assay. Patients presented during infancy or childhood with a variable association of neurodevelopmental disorder, generalized epilepsy, action myoclonus/cortical tremor and ataxia. Later in the disease course, they experienced a slow neurological decline with the emergence of hyperkinetic and/or hypokinetic movement disorder, cognitive deterioration and psychiatric disturbances. Storage of lipidic material and altered lysosomes were detected in myelinated fibres and fibroblasts, suggesting a dysfunction of the lysosomal enzymatic scavenger machinery. Serum glycoprotein hypoglycosylation was not detected and, in contrast to retinitis pigmentosa and other congenital disorders of glycosylation involving dolichol metabolism, the urinary dolichol D18/D19 ratio was normal. Mapping the disease-causing variants into the protein structure revealed that most of them clustered around the active site of the DHDDS subunit. Functional studies using yeast complementation assay and in vitro activity measurements confirmed that these changes affected the catalytic activity of the cis-PTase and showed growth defect in yeast complementation system as compared with the wild-type enzyme and retinitis pigmentosa-associated protein. In conclusion, we characterized a distinctive neurodegenerative disorder due to de novo DHDDS variants, which clinically belongs to the spectrum of genetic progressive encephalopathies with myoclonus. Clinical and biochemical data from this cohort depicted a condition at the intersection of congenital disorders of glycosylation and inherited storage diseases with several features akin to of progressive myoclonus epilepsy such as neuronal ceroid lipofuscinosis and other lysosomal disorders.

In recent years machine learning has made extensive progress in modeling many aspects of mass spectrometry data. We brought together proteomics data generators, repository managers, and machine learning experts in a workshop with the goals to evaluate and explore machine learning applications for realistic modeling of data from multidimensional mass spectrometry-based proteomics analysis of any sample or organism. Following this sample-to-data roadmap helped identify knowledge gaps and define needs. Being able to generate bespoke and realistic synthetic data has legitimate and important uses in system suitability, method development, and algorithm benchmarking, while also posing critical ethical questions. The interdisciplinary nature of the workshop informed discussions of what is currently possible and future opportunities and challenges. In the following perspective we summarize these discussions in the hope of conveying our excitement about the potential of machine learning in proteomics and to inspire future research.

Wound is tissue damage that occurs in the skin. Helichrysum species (Altınotu) are rich in phenolic compounds used in traditional medicine for wound healing. The main component in their flower head (capitulum) is phenolic compounds. The present study investigates the proliferative, oxidative stress, and wound healing properties of the methanolic extract of Helichrysum plicatum subsp. pseudoplicatum capitulum on a human dermal fibroblast (HDF) cell line in this study. H plicatum subsp. pseudoplicatum capitulums were collected in Erzurum, Turkey (altitude 1950 m), dried, pulverized, and extracted with methanol. Firstly, total phenolic contents were determined and secondly, the proliferative effect, oxidative stress activities, and wound healing effects on HDF cells were evaluated by the cell proliferation kit (XTT) test, total antioxidant status (TAS), and total oxidant status (TOS) commercial kits, and the scratch experiment by taking microscopic images of the cells at 0, 12, 18, and 24 h, respectively. Total phenolic content was found to be 142.00 ± 0.73 mg gallic acid equivalent per gram (GAE/g) extract. The capitulum extract has a proliferative effect at 0.5 to 10 µg/mL concentrations according to the XTT test results. It was observed that TAS levels significantly increased in the plant extract at the concentration ranges 1 to 10 µg/mL ( P

In this paper, magnetic-molecularly imprinted polymer was used for the preconcentration of trace levels of imidacloprid in water and apple samples prior to liquid chromatography-quadrupole-time-of-flight mass spectrometric determination. The selectivity of the magnetic polymer was united with the sensitivity and the high resolving power of the chromatographic system. The developed method showed a linear range from 10.0 to 500.0 mu g/L. The quantitative recoveries were obtained for water and apple samples in the range of 92.0%-99.0 %. The relative standard deviations of intra-day and inter-day tests were found to be in the range of 0.8%-1.2% and 1.2%-1.6 %, respectively. In addition, the same magnetic-molecularly imprinted polymer (MMIP) can be used at least ten cycles for the determination of imidacloprid. The preconcentration factor of the method was found to be 2.5, and the total preconcentration procedure can be completed in 1 h. Characterization of synthesised particles were executed with various techniques. Due to its suitable limit of detection, dynamic linear range, sensitivity and selectivity, the developed method seemed to be ideal for the determination and preconcentration of imidacloprid in water and fruit samples., Research Fund of Ege University [18 -EGE-MATAL -003], This work was supported by Research Fund of Ege University for financial support (Grant no: 18 -EGE-MATAL -003).

Aflatoxin contamination of dried figs has been a chronic problem for decades but aflatoxin distribution within the fruit has not yet been revealed. In this study, we conducted aflatoxin analyses separately in the seedless part of dried figs and in fig seed oil. The results showed that both seedless part and seed oil were contaminated with aflatoxins at levels close to the regulatory limits set for products for direct consumption. Effectiveness of various adsorbents in removing aflatoxins from aqueous and oily compartments of the fruit and the effects of these treatments on bioactive compounds and physicochemical characteristics were also investigated.

Summary Background and aims Vitamin B6 is involved in a large spectrum of physiological processes and comprises of the vitamers pyridoxamine (PM), pyridoxal (PL), pyridoxine (PN), and their phosphorylated derivatives including the biological active pyridoxal 5′-phosphate (PLP). While PN toxicity is known to complicate several treatments, PM has shown promise in relation to the treatment of metabolic and age-related diseases by blocking oxidative degradation and scavenging toxic dicarbonyl compounds and reactive oxygen species. We aimed to assess the metabolization of oral PM supplements in a single and three daily dose. Materials and methods We optimized and validated a method for the quantification of the B6 vitamers in plasma and urine using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Five healthy volunteers were recruited to study PM metabolization after a single oral dose of 200 mg PM or a three daily dose of 67 mg PM. A third protocol was implemented as control for dietary intake. Venous blood samples, 24 h urine and fasted second void urine samples were collected. Results After a single oral dose of 200 mg PM, plasma PM increased in the first 3 h to a maximum of 2324 ± 266 nmol/L. While plasma PM levels returned to baseline after ~10 h of PM intake, PLP increased to a maximum of 2787 ± 329 nmol/L and reached a plateau. We found a small increase of PN to a maximum of 13.5 ± 2.1 nmol/L; it was nearly undetectable after ~12 h. With a three daily dose of 67 mg PM we observed an increase and decline of plasma PM, PL, and PN concentrations after each PM intake. PLP showed a similar increase as in the single dose protocol and accumulated over time. Conclusion In this study we showed high plasma levels of PM after oral PM supplementation. We found steadily increasing levels of the biologically active PLP, with minimal formation of PN. The B6 vitamer PM is an interesting supplement as an inhibitor of harmful processes in metabolic diseases and for the treatment of vitamin B6 deficiency. Clinical trial registry The study was approved by the Medical Ethics Committee of Maastricht University (NL) and was registered at ClinicalTrials.gov as NCT02954588.

A solid-phase extraction procedure was optimized to extract 3-fucosyllactose and other human milk oligosaccharides (HMOs) from human milk samples separately, followed by absolute quantitation using high-performance anion-exchange chromatography-pulsed amperometric detection and porous graphitized carbon-liquid chromatography-mass spectrometry, respectively. The approach developed was applied on a pilot sample set of 20 human milk samples and paired infant feces collected at around 1 month postpartum. One-dimensional 1H nuclear magnetic resonance spectroscopy was employed on the same samples to determine the relative levels of fucosylated epitopes and sialylated (Neu5Ac) structural elements. Based on different HMO consumption patterns in the gastrointestinal tract, the infants were assigned to three clusters as follows: complete consumption; specific consumption of non-fucosylated HMOs; and, considerable levels of HMOs still present with consumption showing no specific preference. The consumption of HMOs by infant microbiota also showed structure specificity, with HMO core structures and Neu5Ac(α2-3)-decorated HMOs being most prone to degradation. The degree and position of fucosylation impacted HMO metabolization differently.

Background Common genetic variants of the enzymes and efflux pump involved in tacrolimus disposition have been associated with calcineurin inhibitor nephrotoxicity, but their importance is unclear because of the multifactorial background of renal fibrosis. This study explores the pro-fibrotic response of tacrolimus exposure in relation to the differential capacity for tacrolimus metabolism in proximal tubule cells (PTCs) with a variable (pharmaco)genetic background. Methods PTCs were obtained from protocol allograft biopsies with different combinations of CYP3A5 and ABCB1 variants and were incubated with tacrolimus within the concentration range found in vivo. Gene and protein expression, CYP3A5 and P-glycoprotein function, and tacrolimus metabolites were measured in PTC. Connective tissue growth factor (CTGF) expression was assessed in protocol biopsies of kidney allograft recipients. Results PTCs produce CTGF in response to escalating tacrolimus exposure, which is approximately 2-fold higher in cells with the CYP3A5*1 and ABCB1 TT combination in vitro. Increasing tacrolimus exposure results in relative higher generation of the main tacrolimus metabolite {13-O-desmethyl tacrolimus [M1]} in cells with this same genetic background. Protocol biopsies show a larger increase in in vivo CTGF tissue expression over time in TT vs. CC/CT but was not affected by the CYP3A5 genotype. Conclusions Tacrolimus exposure induces a pro-fibrotic response in a PTC model in function of the donor pharmacogenetic background associated with tacrolimus metabolism. This finding provides a mechanistic insight into the nephrotoxicity associated with tacrolimus treatment and offers opportunities for a tailored immunosuppressive treatment.

Streptomycin (STR) and dihydrostreptomycin (DSTR) are the typically encountered aminoglycoside (AMG) residues in honey. For AMG analysis, studies in literature involve impractical and expensive applications such as ion-pairing chromatography, immunoassays, pre and post column derivatizations, or SPE approaches. Pre -treatments of these methods are toilsome and costly. Herein, one-pot, aqueous in-situ derivatization method was presented as a superior protocol. Time and cost-efficient UHPLC-MS/MS method has been developed, and practical sample preparation was introduced. Satisfactory results were reported in method verification studies. The mean recovery values were 102.6% for STR and 101.3% for DSTR. Average values between 1.5% and 9.9% RSDs were found at intra and inter-day precisions. CC alpha (5.7 and 5.8 mu g/kg) and CC beta (6.2 and 6.4 mu g/kg) values were calculated for STR and DSTR respectively. AMG residues were found in 29 out of 110 analyzed samples using validated method. Described novelty enabled comprehensive analysis in an inexpensive and straightfor-ward manner.

Synthetic dyes are by far the most widely applied colourants in industry. However, environmental and sustainability considerations have led to an increasing efforts to substitute them with safer and more sustainable equivalents. One promising class of alternatives is the natural quinones; these are class of cyclic organic compounds characterized by a saturated (C6) ring that contains two oxygen atoms that are bonded to carbonyls and have sufficient conjugation to show color. Therefore, this study looks at the potential of isolating and applying quinone dye molecules from a sustainable source as a possible replacement for synthetic dyes. It presents an in-depth description of the three main classes of quinoid compounds in terms of their structure, occurrence biogenesis and toxicology. Extraction and purification strategies, as well as analytical methods, are then discussed. Finally, current dyeing applications are summarised. The literature review shows that natural quinone dye compounds are ubiquitous, albeit in moderate quantities, but all have a possibility of enhanced production. They also display better dyeability, stability, brightness and fastness compared to other alternative natural dyes, such as anthocyanins and carotenoids. Furthermore, they are safer for the environment than are many synthetic counterparts. Their extraction, purification and analysis are simple and fast, making them potential substitutes for their synthetic equivalents. Graphic Abstract

Polycyclic aromatic hydrocarbons (PAHs) are considered priority hazardous substances due to their carcinogenic activity and risk to public health. Strict regulations are in place limiting their release into the environment, but enforcement is hampered by a lack of adequate field-testing procedure, instead relying on sending samples to centralised analytical facilities. Reliably monitoring levels of PAHs in the field is a challenge, owing to the lack of field-deployable analytical methods able to separate, identify, and quantify the complex mixtures in which PAHs are typically observed. Here, we report the development of a hand-portable system based on high-performance liquid chromatography incorporating a spectrally wide absorption detector, capable of fingerprinting PAHs based on their characteristic spectral absorption profiles: identifying 100% of the 24 PAHs tested, including full coverage of the United States Environmental Protection Agency priority pollutant list. We report unsupervised methods to exploit these new capabilities for feature detection and identification, robust enough to detect and classify co-eluting and hidden peaks. Identification is fully independent of their characteristic retention times, mitigating matrix effects which can preclude reliable determination of these analytes in challenging samples. We anticipate the platform to enable more sophisticated analytical measurements, supporting real-time decision making in the field. Portable liquid chromatography instruments enable a myriad of applications in field research. Here a handportable system incorporating a broadband absorption detector is used to separate and classify polycyclic aromatic hydrocarbons from environmental water samples based solely on spectral fingerprinting.

The presence of emerging contaminants in the environment, such as pharmaceuticals, is a growing global concern. The excessive use of medication globally, together with the recalcitrance of pharmaceuticals in traditional wastewater treatment systems, has caused these compounds to present a severe environmental problem. In recent years, the increase in their availability, access and use of drugs has caused concentrations in water bodies to rise substantially. Considered as emerging contaminants, pharmaceuticals represent a challenge in the field of environmental remediation; therefore, alternative add-on systems for traditional wastewater treatment plants are continuously being developed to mitigate their impact and reduce their effects on the environment and human health. In this review, we describe the current status and impact of pharmaceutical compounds as emerging contaminants, focusing on their presence in water bodies, and analyzing the development of bioremediation systems, especially mycoremediation, for the removal of these pharmaceutical compounds with a special focus on fungal technologies.

The inclusion of bioaugmented low-cost biochar in current wastewater treatment technologies is a promising way to enhance the removal and degradation of emerging contaminants. In this paper, the properties of two wood waste biochars (wood waste mix - AB, and date palm fiber wood - PDF), and coffee bean husks (COF), produced at four temperatures (350, 450, 500, 550°C) were compared, and investigated in the presence of Geobacter sulfurreducens or a mixed freshwater stream bacterial culture to understand their potential for the adsorption and biotransformation of two types of pesticides (thiacloprid, pirimicarb), and two pharmaceuticals (ibuprofen, diclofenac). Biochar yield was similar for all three biochars and ranged between 30 and 35%. The ash content of PDF and COF was significantly higher than AB. pH and electrical conductivity (EC) were initially high for COF (pH: 7.4–8; EC: 3–4.27 mS/cm) and PDF (pH: 7.7–10.1; EC: 4–6.24 mS/cm) after 24 h, but stabilized at neutral pH and − (550°C), and 846 mg/L sulphate (350°C). Lower pyrolysis temperatures reduced leachable anions. The biochars were highly (ultra)microporous with little meso- and macroporosity. The adsorption experiments showed that AB and COF biochars were both suited to sorb more than 90% of the initially spiked 10 ppm pirimicarb, AB removed 50.2% of the initial diclofenac concentration compared to only 5% for the no-biochar control, and both biochars could remove about 55% of the initially spiked thiacloprid, and 40% of the ibuprofen. In the presence of a mixed culture, on average 30% more thiacloprid and ibuprofen was removed from the supernatant by AB and COF than the sterile control. This work shows that selected wood-waste feedstocks and low pyrolysis temperature can produce environmentally-safe biochars that have suitable characteristics to sorb emergent pollutants from water. These materials could be further studied in multi-pollution sorption/competition experiments, and in larger environmental wastewater treatment systems.

Background Knowledge of therapy-induced intestinal tract concentrations of antimicrobials allows for interpretation and prediction of antimicrobial resistance selection within the intestinal microbiota. This study describes the impact of three different doses of enrofloxacin (ENR) and two different administration routes on the intestinal concentration of ENR and on the fecal Escherichia coli populations in pigs. Enrofloxacin was administered on three consecutive days to four different treatment groups. The groups either received an oral bolus administration of ENR (conventional or half dose) or an intramuscular administration (conventional or double dose). Results Quantitative analysis of fecal samples showed high ENR concentrations in all groups, ranging from 5.114 ± 1.272 μg/g up to 39.54 ± 10.43 μg/g at the end of the treatment period. In addition, analysis of the luminal intestinal content revealed an increase of ENR concentration from the proximal to the distal intestinal tract segments, with no significant effect of administration route. Fecal samples were also screened for resistance in E. coli isolates against ENR. Wild-type (MIC≤0.125 μg/mL) and non-wild-type (0.125 E. coli isolates were found at time 0 h. At the end of treatment (3 days) only non-wild-type isolates (MIC≥32 μg/mL) were found. Conclusions In conclusion, the observed intestinal ENR concentrations in all groups showed to be both theoretically (based on pharmacokinetic and pharmacodynamic principles) and effectively (in vivo measurement) capable of significantly reducing the intestinal E. coli wild-type population.

Citalopram hydrobromide (CP) is a selective serotonin reuptake inhibitor for the treatment of the depressive disorder. In this work, a novel label-free recognition method based on an extended gate field-effect transistor (EGFET) for selective determination of CP is investigated. A Pt electrode covered by a modified PVC membrane, as the sensing part, is connected to the gate of a MOSFET transducer. Two types of PVC membranes containing ion-pairs, CP ion and tetraphenylborate or phosphotungstate ions (with different percentages), are used in this study. The obtained results indicate that the PVC membrane containing 7 % CP-tetraphenylborate has the highest sensitivity, with a detection limit of 10-12 M. The sensor exhibits two linear response ranges, in the ranges of 10-11 - 10-7 M and 10-6 - 10-2 M, characterized by different sensitivities. The proposed method shows clear advantages in terms of increased accuracy, sensitivity, precision, and selectivity. The sensor is successfully applied to assay citalopram in tablets. The features of simplicity in preparation, fast response, capability for the miniaturization, and easy operatory, make the sensor a promising candidate for future integrated lab-on-chip devices for pharmaceutical quality control measurements.

Cost-efficient deterministic method of moments solvers, as widely used to calculate average characteristics of chemical processes driven by population variations (e.g., average chain lengths), can be a posteriori extended with approximated solutions delivering distributed properties (e.g., chain length distributions). However, these solutions are rarely verified, specifically for complex systems with many population members and strong coupling, as is the case for industrially relevant free-radical-induced grafting (FRIG) toward graft copolymer (GC) synthesis with monomer unit dependent reactions. FRIG, as studied in the present work with polybutadiene at low styrene conversions, is an important chemical process, for example, the production of compatibilizers and high-impact materials. Deterministic model validation is uniquely performed by benchmarking the low to medium molar mass (MM) results (29 topologies) in the log-molar mass distribution with detailed matrix-based kinetic Monte Carlo simulation output, inherently capable of mapping distributions. The GC product is identified to be a heterogeneous mixture in MM, chemical composition, and molecular topology at any styrene conversion. The molecular structural evolution during GC synthesis is further theoretically related to both one-dimensional size-exclusion chromatography (1D-SEC) and two-dimensional liquid chromatography (2D-LC) analysis. It is shown that conventional SEC-even in the absence of broadening-is insufficient for GC separation, mainly due to the unavoidable coelution of topologically different GC species. In any case, the parallel running of advanced modeling tools allows for detailed molecular interpretation.

The origin of the peak skewness that can be observed when applying the deconvolution method to isolate the diffusion process from the flow processes for peak parking experiments conducted under conditions of slow radial equilibration and strong trans-column velocity gradients was investigated. Numerical simulations were carried out for a variety of trans-column velocity profiles and a broad range of experimental conditions and system parameters were investigated. Results show that, under the aforementioned conditions, the traditionally employed variance subtraction method displays a consistent error which follows the dynamics of the diffusive relaxation during both the peak parking and the flow steps. It is also found that, under the same conditions, the peak deconvolution method is bound to produce deconvoluted "parking-only" peaks that are strongly asymmetric, despite the perfectly symmetric nature of the pure diffusion process marking this parking step. It is shown that this asymmetry is acquired during the flow step following the parking stop. During this step, parked and non-parked peaks are deformed in different ways, despite being subjected to the same trans-column velocity profile. This different deformation cannot be filtered away with the deconvolution or the variance subtraction method, hence introducing an error. Solutions to alleviate the peak skewness and the variance error consist of parking the peak close to the inlet or the outlet or exiting the parked peak through the column inlet (flow reversal method). Under the considered conditions, these approaches could reduce the error on the measured effective diffusion coefficient up to 87%. Carrying out the variance subtraction or the deconvolution process with a peak that has also been parked for a substantially long parking time instead of using a "no-parking" peak as is customary done, is another option to counter the effect. ispartof: JOURNAL OF CHROMATOGRAPHY A vol:1656 ispartof: location:Netherlands status: published

Nivolumab (Opdivo®) is a fully human immunoglobulin G4 isotype approved for the treatment of many cancers. It acts as an immune checkpoint inhibitor by blocking the interaction between PD-1 (Programmed Cell Death Protein 1) – an inhibitory receptor expressed on activated T cells- and its ligands, PD-L1 and PD-L2. The quantification of therapeutic proteins in their medicines and pharmaceutical preparations remains challenging because the protein content, a critical quality attribute, must be rigorously calculated using a validated stabilityindicating method, such as that indicated by the International Conference on Harmonization (ICH) quality guidelines, and this requires the analysis of the drug in the presence of its degraded products. In this work, we present an strategy based on the combined use of the UV and MS data to full file the requirement of the ICH-Q2 (R1) to develop and validated as stability indicated a (RP)UHPLC/UV-(HESI/Orbitrap™)MS method for the quantification of nivolumab in medicinal products. A comparative study of all figures of merit of the method using UV or MS data are shown and discussed. The results show that linearity was similar for the two detectors and was established over a range of 4–45 μg/mL and 1–45 μg/mL for the UV and (HESI/Orbitrap™)MS signals, respectively. The sensitivity of the method was higher when using the (HESI/Orbitrap™)MS signal (0.2 μg/mL) than with the UV(2.0 μg/mL). However, the UV signal provided better accuracy and precision than the (HESI/ Orbitrap™)MS signal, which did not meet the criteria for method robustness and system suitability. In spite of this, the MS signal plays a crucial role in this methodology by obtaining the molecular weight profile of the nivolumab isoforms, so enabling us to propose the glycans profile and detect structural modification due to degradation. The specificity of the method was evaluated by conducting forced degradation tests on samples of nivolumab in medicine form. The aim was to find out whether nivolumab suffers structural modifications when subject to stress. Structural modifications were detected by analysing the MS isoform profile, as changes of this kind promote new isoforms that are not chromatographically separated or detected by the UV signal. In this way, we demonstrated that the (RP)UHPLC/UV-(HESI/Orbitrap™)MS method was capable of detecting nivolumab degradation, and was suitable for use in nivolumab stability studies. Thus, the protein content in the daily surplus of the Opdivo® medicine, stored either at room temperature (20 ◦C) or refrigerated at 4 ◦C, could be tracked for 15 days., FPU18/03131 Ministry of Universities, Spain, (P20_01029) Junta de Andalucía (Spain) and European Regional Development Funds, postdoctoral position from the Junta de Andalucía, Spain, Project P20-01029 (I + D + i - Junta de Andalucía, Spain) Project B-FQM-308-UGR20 (Universidad de Granada, Proyectos I + D + i del Programa Operativo FEDER Andalucía 2020), CBUA/ Universidad de Granada

Human milk oligosaccharides (hMOS) are associated with health benefits for newborns. We studied the composition of goat MOS (gMOS) from colostrum up to the 9th month of lactation to conceive an overview of the structures present and their fate. Potential correlations with factors such as age, parity, and lifetime milk production were examined. An effective method for gMOS extraction and ultra-high-performance liquid chromatography coupled to fluorescence detection (UPLC-FLD) analysis was established, following 2-aminobenzamide gMOS labeling. Considerable biological variability was highlighted among the 12 quantified gMOS and the 9 non-quantified structures in the individual milk samples. Most characteristic, 2'-fucosyllactose was present in 73.7% of the milk samples analyzed, suggesting the possibility of a secretor/non-secretor goat genotype, similar to humans. Contributing factors to the observed biological variability were goat age, parity, lifetime milk production, and the kids' sex. The results significantly contribute to the current understanding of (variations in) gMOS composition.

Publisher Copyright: © 2022. The Author(s). A genome-wide association study identifies MFGE8 as protective against coronary atherosclerosis in European and East Asian populations. Cardiovascular diseases are the leading cause of premature death and disability worldwide, with both genetic and environmental determinants. While genome-wide association studies have identified multiple genetic loci associated with cardiovascular diseases, exact genes driving these associations remain mostly uncovered. Due to Finland's population history, many deleterious and high-impact variants are enriched in the Finnish population giving a possibility to find genetic associations for protein-truncating variants that likely tie the association to a gene and that would not be detected elsewhere. In a large Finnish biobank study FinnGen, we identified an association between an inframe insertion rs534125149 in MFGE8 (encoding lactadherin) and protection against coronary atherosclerosis. This variant is highly enriched in Finland, and the protective association was replicated in meta-analysis of BioBank Japan and Estonian biobank. Additionally, we identified a protective association between splice acceptor variant rs201988637 in MFGE8 and coronary atherosclerosis, independent of the rs534125149, with no significant risk-increasing associations. This variant was also associated with lower pulse pressure, pointing towards a function of MFGE8 in arterial aging also in humans in addition to previous evidence in mice. In conclusion, our results suggest that inhibiting the production of lactadherin could lower the risk for coronary heart disease substantially.