Mass Spectrometry Research Articles

Characterization of lamp oil residues from Tangchaodun Site, Xinjiang, northwest China during the 8th−13th centuries CE

Lamp illuminants reflect how ancient people exploited natural resources, such as animal fat, plant oil and mineral oil. Besides, the commercial trade of lamp oil, including olive oil, represented economic exchange. In addition, oil lamps were often used in religious practices to glorify deities and illuminate grand halls. Although many ancient lamps have been found in China, there are few scientific reports regarding ancient lamp illuminants. In this study, chemical analysis including FTIR (Fourier Transform Infrared Spectroscopy), Py-GC/MS (Pyrolysis Gas Chromatography-Mass Spectrometry), GC–MS (Gas Chromatography-Mass Spectrometry), and GC-C-IRMS (Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry) was applied to characterize black residues remaining on the surface of pottery lamps from the Tangchaodun site, northwest China, from the 8th to 13th centuries CE. The results show that the black residues are soot, and both ruminant adipose fat and plant oil were used as the primary fuel. These lamps share similar morphology and utilize similar combustibles, implying that the production of both lamps and oil may have undergone convergent evolution driven by aesthetic preferences. Furthermore, the ruminant animals mainly fed on C4 plants, indicating that these animals were raised in pens and millet agriculture was practiced in this region.

Discovery of bitter masking compounds from Allspice (Pimenta dioica) using sensory guided isolation

Bitter substances in functional foods and beverages can act as nutraceuticals, offering potential health benefits. However, their unpleasant sensory impact reduces the consumption of these foods. Consequently, the discovery of bitter masking compounds is crucial for enhancing the intake of bioactive compounds in functional foods and beverages. Bitter taste is mediated by TAS2Rs, a sub-family of G-protein-coupled receptors. TAS2R14 is especially pivotal in the perception of bitterness, as it is one of the most broadly tuned bitter receptors. In this study, allspice was extracted and purified to yield five single compounds based on sensory guided fractionation. The structures of each compound were determined based on nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HR-MS). In a sensory evaluation, compound 1 exhibited bitter masking activity against quinine. Molecular docking analysis revealed that compound 1 could act as an antagonist of the TAS2R14 bitter receptor.

Determination of vancomycin and meropenem in serum and synovial fluid of patients with prosthetic joint infections using UPLC-MS/MS.

Numerous studies have suggested that intra-articular administration of antibiotics following primary revision surgery may be one of the methods for treating prosthetic joint infection (PJI). Vancomycin and meropenem are the two most commonly used antibiotics for local application. Determining the concentrations of vancomycin and meropenem in the serum and synovial fluid of patients with PJI plays a significant role in further optimizing local medication schemes and effectively eradicating biofilm infections. This study aimed to establish a rapid, sensitive, and accurate ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determining the concentrations of vancomycin and meropenem in human serum and synovial fluid. Serum samples were processed using acetonitrile precipitation of proteins and dichloromethane extraction, while synovial fluid samples were diluted before analysis. Chromatographic separation was achieved in 6min on a Waters Acquity UPLC BEH C18 column, with the mobile phase consisting of 0.1% formic acid in water (solvent A) and acetonitrile (solvent B). Quantification was carried out using a Waters XEVO TQD triple quadrupole mass spectrometer with an electrospray ionization (ESI) source in positive ion mode. The multiple reaction monitoring (MRM) mode was employed to detect the following quantifier ion transitions: 717.95-99.97 (norvancomycin), 725.90-100.04 (vancomycin), 384.16-67.99 (meropenem). The method validation conformed to the guidelines of the FDA and the Chinese Pharmacopoeia. The method demonstrated good linearity within the range of 0.5-50 μg/ml for serum and 0.5-100 μg/ml for synovial fluid. Selectivity, intra-day and inter-day precision and accuracy, extraction recovery, matrix effect, and stability validation results all met the required standards. This method has been successfully applied in the pharmacokinetic/pharmacodynamic (PK/PD) studies of patients with PJI.

Efficacy and safety of sympathetic mapping and ablation of renal nerves for the treatment of hypertension (SMART): 6-month follow-up of a randomised, controlled trial

Previous trials of renal denervation (RDN) have been designed to investigate reduction of blood pressure (BP) as the primary efficacy endpoint using non-selective RDN without intraoperatively verified RDN success. It is an unmet clinical need to map renal nerves, selectively denervate renal sympathetic nerves, provide readouts for the interventionalists and avoid futile RDN. We aimed to examine the safety and efficacy of renal nerve mapping/selective renal denervation (msRDN) in patients with uncontrolled hypertension (HTN) and determine whether antihypertensive drug burden is reduced while office systolic BP (OSBP) is controlled to target level (＜140mmHg). We conducted a randomized, prospective, multicenter, single-blinded, sham-controlled trial. The study combined two efficacy endpoints at 6 months as primary outcomes: The control rate of patients with OSBP ＜140mmHg (non-inferior outcome) and change in the composite index of antihypertensive drugs (Drug Index) in the treatment versus Sham group (superior outcome). This design avoids confounding from excess drug-taking in the Sham group. Antihypertensive drug burden was assessed by a composite index constructed as: Class N (number of classes of antihypertensive drugs)×(sum of doses). 15 hospitals in China participated in the study and 220 patients were enrolled in a 1:1 ratio (msRDN vs Sham). The key inclusion criteria included: age (18-65 years old), history of essential HTN (at least 6 months), heart rate (≥70 bpm), OSBP (≥150mmHg and ≤180mmHg), ambulatory BP monitoring (ABPM, 24-h SBP ≥130mmHg or daytime SBP ≥135mmHg or nighttime SBP ≥120mmHg), renal artery stenosis (＜50%) and renal function (eGFR ＞45mL/min/1.73m2). The catheter with both stimulation and ablation functions was inserted in the distal renal main artery. The RDN site (hot spot) was selected if SBP increased (≥5mmHg) by intra-renal artery (RA) electrical stimulation; an adequate RDN was confirmed by repeated electronic stimulation if no increase in BP otherwise, a 2nd ablation was performed at the same site. At sites where there was decreased SBP (≥5mmHg, cold spot) or no BP response (neutral spot) to stimulation, no ablation was performed. The mapping, ablation and confirmation procedure was repeated until the entire renal main artery had been tested then either treated or avoided. AftermsRDN, patients had to follow a predefined, vigorous drug titration regimen in order to achieve target OSBP (＜140mmHg). Drug adherence was monitored by liquid chromatography-tandem mass spectrometry analysis using urine. This study is registered with ClinicalTrials.gov (NCT02761811) and 5-year follow-up is ongoing. Between July 8, 2016 and February 23, 2022, 611 patients were consented, 220 patients were enrolled in the study who received standardized antihypertensive drug treatments (at least two drugs) for at least 28 days, presented OSBP ≥150mmHg and ≤180mmHg and metall inclusion and exclusion criteria. In left RA and right RA, mapped sites were 8.2 (3.0) and 8.0 (2.7), hot/ablated sites were 3.7 (1.4) and 4.0 (1.6), cold spots were 2.4 (2.6) and 2.0 (2.2), neutral spots were 2.0 (2.1) and 2.0 (2.1), respectively. Hot, cold and neutral spots was 48.0%, 27.5% and 24.4% of total mapped sites, respectively. At 6M, the Control Rate of OSBP was comparable between msRDN and Sham group (95.4% vs 92.8%, p=0.429), achieved non-inferiority margin-10% (2.69%; 95% CI-4.11%, 9.83%, p ＜ 0.001 for non-inferiority); the change in Drug Index was significantly lower in msRDN group compared to Sham group (4.37 (6.65) vs 7.61 (10.31), p=0.010) and superior to Sham group (-3.25; 95% CI-5.56,-0.94, p=0.003), indicating msRDN patients need significantly fewer drugs to control OSBP ＜140mmHg. 24-hour ambulatory SBP decreased from 146.8 (13.9) mmHg by 10.8 (14.1) mmHg, and from 149.8 (12.8) mmHg by 10.0 (14.0) mmHg in msRDN and Sham groups, respectively (p<0.001 from Baseline; p>0.05 between groups). Safety profiles were comparable between msRDN and Sham groups, demonstrating the safety and efficacy of renal mapping/selective RDN to treat uncontrolled HTN. The msRDN therapy achieved the goals of reducing the drug burden of HTN patients and controlling OSBP <140mmHg, with only approximately four targeted ablations per renal main artery, much lower than in previous trials. SyMap Medical (Suzhou), LTD, Suzhou, China.

Dynamic Behavior of Systemic Insecticide Residues in Snapdragons

Pesticide application is used in horticulture to reduce plant damage from organisms such as insects and mites. Systemic insecticides are highly efficacious and readily taken up by plant tissues. However, pesticide-treated plants may impose risks to nontarget insects or other organisms within ecosystems. In this study, insecticide residues in nectar, leaves, and flower petals of the horticulturally significant herbaceous annual snapdragon, Antirrhinum majus (Lamiales: Plantaginaceae), were assessed at two locations over several weeks following foliar and drench treatment with five systemic insecticides. Concentrations of the insecticides were determined by liquid chromatography–mass spectrometry. The independent effects Application Method, Application Rate, and Time were statistically significant among all active ingredients in the three matrices in both sites in California (CA) and New Jersey (NJ). The interaction effects were also generally statistically significant in the CA site but less consistently so in the NJ site, dependent on the active ingredient and matrix. Post hoc analyses found the highest residue concentrations in leaves and the lowest in nectar, a trend generally consistent over time regardless of active ingredient for both the CA and NJ sites. The results of this study are discussed in the context of conserving pollinators and other beneficial insects. It is recommended that similar studies should be implemented in different geographical regions and climates, along with multiyear studies for perennial ornamental plants.

Accelerator mass spectrometry measurements of 233U in groundwater, soil and vegetation at a legacy radioactive waste site

Low-level radioactive wastes were disposed at the Little Forest Legacy Site (LFLS) near Sydney, Australia between 1960 and 1968. According to the disposal records, 233U contributes a significant portion of the inventory of actinide activity buried in the LFLS trenches. Although the presence of 233U in environmental samples from LFLS has been previously inferred from alpha-spectrometry measurements, it has been difficult to quantify because the 233U and 234U α-peaks are superimposed. Therefore, the amounts of 233U in groundwaters, soils and vegetation from the vicinity of the LFLS were measured using accelerator mass spectrometry (AMS). The AMS results show the presence of 233U in numerous environmental samples, particularly those obtained within, and in the immediate vicinity of, the trenched area. There is evidence for dispersion of 233U in groundwater (possibly mobilised by co-disposed organic liquids), and the data also suggest other sources of 233U contamination in addition to the trench wastes. These may include leakages and spills from waste drums as well as waste burnings, which also occurred at the site. The AMS results confirm the historic information regarding disposal of 233U in the LFLS trenches. The AMS technique has been valuable to ascertain the distribution and environmental behaviour of 233U at the LFLS and the results demonstrate the applicability of AMS for evaluating contamination of 233U at other radioactive waste sites.

On the fate of protonated chloroformates in the gas phase: a competition between forming HCl and chloroformic acid.

Chloroformates are prevalent in the atmosphere due to their utilization as fuel additives and industrial solvents. These species may undergo interactions with atmospheric water resulting in protonated chloroformates. This study delves into the gas-phase dissociation of these protonated species. Tandem mass spectrometry was employed to scrutinize the unimolecular dissociation of protonated methyl (1), ethyl (2), neopentyl (3), and phenyl chloroformate (4). Notably, 1 and 4 exhibited HCl loss, yielding CH3OCO+ and C6H5OCO+, respectively, with 1 additionally generating neutral methanol and ClCO+. 4 additionally loses CO and CO2. In contrast, 2 and 3 each only exhibit a single fragmentation channel, with 2 losing C2H4 to generate protonated chloroformic acid and 3 generating protonated 2-methylbutene by losing neutral chloroformic acid. Density functional theory at the B3LYP/6-311+G(d,p) level of theory was employed to explore minimum energy reaction pathways for each ion, and CBS-QB3 single-point energy calculations were added to provide reliable energetics. The Rice-Ramsperger-Kassel-Marcus (RRKM) calculations of the rate constants for selected competing processes were carried out to link theory and experiment. One common unimolecular process observed was the 1,3-H shift of the proton from the carbonyl oxygen to the ester oxygen before dissociation.

Analysis of Blackening Reaction of Zn–Mg–Al Alloy Coated Steel Prepared by Anodizing Process

The rising demand for black-treated steel faces challenges with conventional black painting due to issues like scratching and peeling, impacting corrosion resistance and aesthetics. This study explores an alternative method, anodic oxidation, to blacken the surfaces of galvanized or coated steel plates. Parameters like temperature, duration, current density, and gas type were varied during the blackening process. The investigation aimed to identify key factors influencing the blackening. Scanning electron microscopy observed the morphology, while energy-dispersive X-ray spectroscopy and glow discharge mass spectrometry analyzed the chemical composition distribution. X-ray diffraction and X-ray photoelectron spectroscopy conducted compound crystal structure analysis. Results indicate higher temperatures, longer durations, and higher current densities improve blackening through anodic oxidation. Increased magnesium proportion on the surface leads to roughness and porous magnesium oxide formation, enhancing light absorption and explaining the observed blackening effect.

Investigation of the Molecular Mechanism Underlying the Therapeutic Effect of Perilla frutescens L. Essential Oil on Acute Lung Injury Using Gas Chromatography Mass Spectrometry and Network Pharmacology.

The present study aimed to investigate the molecular mechanism through which Perilla essential oil treats acute lung injury (ALI) through network pharmacology, molecular docking, and in vitro assays. Relevant ALI targets of the active ingredients of Perilla essential oil were predicted using the SwissTargetPrediction database and meta TarFisher database. These ALI targets were then screened using GeneCards and DisGeNET, and differentially expressed ALI target genes were identified using the Gene Expression Omnibus (GEO) database. Next, key targets were enriched using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Protein-protein interaction network analysis was performed to obtain targets with the highest degree values for molecular docking with Perilla essential oil active ingredients. For in vitro experiments, lipopolysaccharide (LPS) was used to induce an ALI inflammation model using RAW264.7 cells. The model cells were then treated with Perilla essential oil to detect the protein expression levels of vascular endothelial factor (NO), tumor necrosis factor (TNF-α), and p65 nuclear transcription factor in them. Sixty-eight key targets of Perilla oil were identified for the treatment of ALI. These targets were found to be involved in biological processes related to peptides, response to lipopolysaccharides, the positive regulation of cytokine production, etc., using GO. The signaling pathways found to be associated with the targets included the AGE-RAGE signaling pathway in diabetic complications, the NF-kappa B signaling pathway, and small cell lung cancer and other inflammatory signaling pathways. The five key targets that showed good binding activity with Perilla oil active ingredients included TNF, RELA, PARP1, PTGS2, and IRAK4. In vitro assays showed that Perilla essential oil could significantly reduce NO and TNF-α levels and inhibit the phosphorylation of nuclear transcription factor P65, thus inhibiting the activation of NF-κB signaling pathway. Conclusion Perilla essential oil can play a role in the treatment of ALI by inhibiting the activation of the NF-κB signaling pathway and preventing an excessive inflammatory response. This study thus provides a reference for the in-depth study of the mechanisms through which Perilla essential oil treats ALI.

Integrative multiomics analysis identifies molecular subtypes and potential targets of hepatocellular carcinoma.

The liver is anatomically divided into eight segments based on the distribution of Glisson's triad. However, the molecular mechanisms underlying each segment and its association with hepatocellular carcinoma (HCC) heterogeneity are not well understood. In this study, our objective is to conduct a comprehensive multiomics profiling of the segmentation atlas in order to investigate potential subtypes and therapeutic approaches for HCC. A high throughput liquid chromatography-tandem mass spectrometer strategy was employed to comprehensively analyse proteome, lipidome and metabolome data, with a focus on segment-resolved multiomics profiling. To classify HCC subtypes, the obtained data with normal reference profiling were integrated. Additionally, potential therapeutic targets for HCC were identified using immunohistochemistry assays. The effectiveness of these targets were further validated through patient-derived organoid (PDO) assays. A multiomics profiling of 8536 high-confidence proteins, 1029 polar metabolites and 3381 nonredundant lipids was performed to analyse the segmentation atlas of HCC. The analysis of the data revealed that in normal adjacent tissues, the left lobe was primarily involved in energy metabolism, while the right lobe was associated with small molecule metabolism. Based on the normal reference atlas, HCC patients with segment-resolved classification were divided into three subtypes. The C1 subtype showed enrichment in ribosome biogenesis, the C2 subtype exhibited an intermediate phenotype, while the C3 subtype was closely associated with neutrophil degranulation. Furthermore, using the PDO assay, exportin 1 (XPO1) and 5-lipoxygenase (ALOX5) were identified as potential targets for the C1 and C3 subtypes, respectively. Our extensive analysis of the segmentation atlas in multiomics profiling defines molecular subtypes of HCC and uncovers potential therapeutic strategies that have the potential to enhance the prognosis of HCC.

Study on Sustainable Resource Utilization as Ligand Precursors in Coordination Chemistry

Ligand Precursors in Coordination Chemistry.” Sustainable resource utilization in ligand precursor synthesis is crucial for advancing green chemistry principles and addressing environmental concerns in coordination chemistry. This study investigates the feasibility and efficacy of utilizing renewable resources as precursor materials for ligand synthesis, aiming to reduce environmental impact while maintaining the efficacy and versatility required in coordination chemistry applications. Through a combination of synthetic organic chemistry and coordination chemistry techniques, novel ligand precursors derived from sustainable sources were synthesized and characterized. Various renewable feedstocks, including biomass-derived compounds and waste materials, were explored as potential starting materials for ligand precursor synthesis. Optimization of synthetic routes and reaction conditions led to the development of efficient protocols for the synthesis of sustainable ligand precursors. Characterization techniques such as nuclear magnetic resonance spectroscopy, mass spectrometry, and infrared spectroscopy were employed to confirm the structures and purities of synthesized ligand precursors. Evaluation of the performance of sustainable ligand precursors in coordination chemistry reactions demonstrated their ability to form stable complexes with metal ions and exhibit catalytic activity in relevant reactions. Comparative studies with traditional ligands revealed the environmental advantages of sustainable ligand precursors, including reduced energy consumption, greenhouse gas emissions, and solvent usage. Overall, this study highlights the importance of sustainable resource utilization in ligand precursor synthesis for promoting environmentally friendly practices in coordination chemistry and advancing the principles of green chemistry.

Quantitative and qualitative phytochemical analysis of ethanol leaf extract of Pleiocarpa mutica

This research project examined Pleiocarpa mutica leaf extract, extracted from ethanol using both quantitative and qualitative phytochemical analysis. Sourced is at Ugbene-Ajima in the Uzo-Uwani Local Government Area of Enugu State, Nigeria, where fresh Pleiocarpa mutica leaves were collected. After been gathered, the newly sprouted P. mutica leaves were cleaned. We then shade-dried the leaves until they were crispy, rotating them frequently to prevent them from rotting. A mechanical grinder was utilized to reduce the dried leaves to a powdered state while a maceration flask was used to macerate 1.5 kg of the known weight of the ground leaves in 10 liters of 100% ethanol. In using a muslin cloth, the mixture was filtered into a flask with a flat bottom after being left for 72 hours with irregular stirring. Quantitative phytochemical analysis procedures identified the specific concentration of a certain chemical constituent present in a plant sample using methods such as mass spectrometry, chromatography, and spectrophotometry. This makes it possible to determine the concentration of a component in the plant material. Qualitative phytochemical analysis techniques focus on identifying the presence or absence of several chemical groups in a plant sample. Discovering the existence of substances such as alkaloids, phenols, terpenoids, and flavonoids often requires a range of chemical analyses or the use of specific reagents. Thus, the standard quantitative and qualitative phytochemical analysis technique were used to investigate the chemical composition of plants and identify bioactive compounds that may offer nutritional, medicinal, or pharmacological advantages. Hence, it has been shown that the ethanol leaves extract of P. mutica contains various concentrations of phytochemical constituents, which might be responsible for its biological activities.

The Assessment of Heat-treated Turkish Type Dry Fermented Sausage (Sucuk) with Some Mixed Probiotic Lactic Acid Bacteria

T his study aims to obtain a combination of probiotic strains resistant to stress conditions, which good effect the qualitative properties of heat-treated sausage. Lactic acid bacteria (LAB) strains were identified by matrix-assisted laser desorption-ionization-time of flight mass spectrometry (MALDI-TOF MS) after phenotypic evaluation and evaluated for probiotic potential including acid and bile salt tolerance, antibiotic resistance, and antibacterial activity. All tested Lactiplantibacillus plantarum strains showed high antibacterial activity. L. plantarum (PeE2) and additionally two Enterococcus strains with the best probiotic properties were selected among the strains. Except for the control group, 6 different sucuk doughs were prepared using them as mixed cultures. The ripened sucuk doughs (18 h, 4 ºC) were subjected to fast-drying (30 min, 60 ºC), heat-treated (64 ºC of inner temperature) and rapid-drying (10 min, 75 ºC) stages. The quality characteristics of samples were determined during storage (days 7, 14, and 21). Coliform bacteria growth ≤1 log cfu/g determined on the final day of storage. The lowest TBARS value was determined in L. plantarum +E. faecium (1:1 ratio) mixed culture. Regarding L* and a* color values, lower results were observed in the control group during storage. Using probiotic culture significantly affected textural values, taste and general acceptability (p&lt;0.05) and contributed to the quality parameters. The result of the study may support the development of a new approach for using new probiotics in meat products.

Protein homeostasis maintained by HOOK1 levels promotes the tumorigenic and stemness properties of ovarian cancer cells through reticulum stress and autophagy

BackgroundOvarian cancer has a high mortality rate mainly due to its resistance to currently used therapies. This resistance has been associated with the presence of cancer stem cells (CSCs), interactions with the microenvironment, and intratumoral heterogeneity. Therefore, the search for new therapeutic targets, particularly those targeting CSCs, is important for improving patient prognosis. HOOK1 has been found to be transcriptionally altered in a substantial percentage of ovarian tumors, but its role in tumor initiation and development is still not fully understood.MethodsThe downregulation of HOOK1 was performed in ovarian cancer cell lines using CRISPR/Cas9 technology, followed by growth in vitro and in vivo assays. Subsequently, migration (Boyden chamber), cell death (Western-Blot and flow cytometry) and stemness properties (clonal heterogeneity analysis, tumorspheres assay and flow cytometry) of the downregulated cell lines were analysed. To gain insights into the specific mechanisms of action of HOOK1 in ovarian cancer, a proteomic analysis was performed, followed by Western-blot and cytotoxicity assays to confirm the results found within the mass spectrometry. Immunofluorescence staining, Western-blotting and flow cytometry were also employed to finish uncovering the role of HOOK1 in ovarian cancer.ResultsIn this study, we observed that reducing the levels of HOOK1 in ovarian cancer cells reduced in vitro growth and migration and prevented tumor formation in vivo. Furthermore, HOOK1 reduction led to a decrease in stem-like capabilities in these cells, which, however, did not seem related to the expression of genes traditionally associated with this phenotype. A proteome study, along with other analysis, showed that the downregulation of HOOK1 also induced an increase in endoplasmic reticulum stress levels in these cells. Finally, the decrease in stem-like properties observed in cells with downregulated HOOK1 could be explained by an increase in cell death in the CSC population within the culture due to endoplasmic reticulum stress by the unfolded protein response.ConclusionHOOK1 contributes to maintaining the tumorigenic and stemness properties of ovarian cancer cells by preserving protein homeostasis and could be considered an alternative therapeutic target, especially in combination with inducers of endoplasmic reticulum or proteotoxic stress such as proteasome inhibitors.

Influence of composite chloride salt retardant on the characteristics of coal spontaneous combustion and comprehensive evaluation methodology

ABSTRACT In this study, we aimed to improve the efficacy and longevity of single inhibitors. NaCl, CaCl2, MgCl2, and layered double hydroxides (LDHs) were selected as the inhibitory components for experimental study. The impact of the composite inhibitors on the spontaneous combustion characteristics of the coal was studied through temperature programmed experiment, thermal analysis experiments, thermogravimetric mass spectrometry and entropy weight method, and the optimal inhibitor was determined. The findings indicate that the apparent activation energy (E a ) of the coal sample, with the addition of inhibitors during the oxidation stage, was improved. E a of MgCl2 +1.0 LDHs + Coal was the highest (127.28 kJ/mol), i.e. 35.99 kJ/mol higher than that of the original coal. The inclusion of the inhibitor increased the characteristic ignition temperatures of the coal samples compared with the original coal. The highest ignition temperature was observed for MgCl2 +0.5 LDHs + Coal, reaching (323.98°C), i.e. 11.42°C higher than that of the original coal. The addition of inhibitors significantly inhibited the production of CO. Among the three chloride salts, MgCl2 exhibited the strongest inhibitory effect on CO production. According to the entropy weight method, MgCl2 +0.5 LDHs showed the best inhibitory effect. The results show that the composite halogen inhibitor can effectively inhibit the spontaneous combustion of coal.

Identification and characterization of zinc importers in Corynebacterium diphtheriae.

Corynebacterium diphtheriae is the causative agent of diphtheria, a severe respiratory disease in humans. C. diphtheriae colonizes the human upper respiratory tract, where it acquires zinc, an essential metal required for survival in the host. While the mechanisms for zinc transport by C. diphtheriae are not well characterized, four putative zinc ABC-type transporter loci were recently identified in strain 1737: iutABCD/E (iut), znuACB (znu), nikABCD1 (nik1), and nikABCD2 (nik2). A mutant deleted for all four loci (Δ4) exhibited similar growth to that of the wild-type strain in a zinc-limited medium, suggesting there are additional zinc transporters. Two additional gene loci predicted to be associated with metal import, mntABCD (mnt) and sidAB (sid), were deleted in the Δ4 mutant to construct a new mutant designated Δ6. The C. diphtheriae Δ6 mutant exhibited significantly reduced growth under zinc limitation relative to the wild type, suggesting a deficiency in zinc acquisition. Strains retaining the iut, znu, mnt, or sid loci grew to near-wild-type levels in the absence of the other five loci, indicating that each of these transporters may be involved in zinc uptake. Plasmid complementation with cloned iut, znu, mnt, or nik1 loci also enhanced the growth of the Δ6 mutant. Quantification of intracellular zinc content by inductively coupled plasma mass spectrometry was consistent with reduced zinc uptake by Δ6 relative to the wild type and further supports a zinc uptake function for the transporters encoded by iut, znu, and mnt. This study demonstrates that C. diphtheriae zinc transport is complex and involves multiple zinc uptake systems.IMPORTANCEZinc is a critical nutrient for all forms of life, including human bacterial pathogens. Thus, the tools that bacteria use to acquire zinc from host sources are crucial for pathogenesis. While potential candidates for zinc importers have been identified in Corynebacterium diphtheriae from gene expression studies, to date, no study has clearly demonstrated this function for any of the putative transporters. We show that C. diphtheriae encodes at least six loci associated with zinc import, underscoring the extent of redundancy for zinc acquisition. Furthermore, we provide evidence that a previously studied manganese-regulated importer can also function in zinc import. This study builds upon our knowledge of bacterial zinc transport mechanisms and identifies potential targets for future diphtheria vaccine candidates.

Does in vitro hemolysis affect measurements of plasma apixaban concentration by UPLC-MS and anti-Xa assay?

Hemolytic interference may impact various laboratory tests, including coagulation analyses. Apixaban is the most commonly used direct oral anticoagulant in Norway, and there is lacking knowledge on how apixaban concentration measurements might be influenced by hemolysis. Moreover, hemolysis-induced alterations in apixaban levels could potentially impact the risk of bleeding in specific clinical scenarios. We wanted to study whether hemolysis would increase apixaban concentration and investigate the impact of hemolytic interference on apixaban concentration measurements. Blood samples from 20 apixaban-treated patients and 8 healthy controls were hemolyzed in vitro by a freeze method. The degree of hemolysis was measured with plasma free hemoglobin (PfHb) at baseline and two levels of hemolysis. Apixaban concentration was measured in plasma using both the chromogenic anti-Xa method and the ultraperformance liquid chromatography mass spectrometry (UPLC-MS). Thrombin generation assay was performed to assess coagulability. UPLC-MS measurements showed a mean concentration change of -1.66% (±3.2%, p = 0.005) and anti-Xa assay showed a mean concentration change of 3.37% (±6.5%, p = 0.09) with increasing hemolysis. Thrombin generation lagtime decreased, and endogenous thrombin potential and peak thrombin increased with increasing hemolysis in both the control group and the apixaban group. Apixaban concentration measurements by anti-Xa assay and UPLC-MS were not affected by hemolysis to a clinically relevant extent. Furthermore, hemolysis did not lead to hypocoagulability when assessed by thrombin generation.

Integration of protein L-immobilized epoxy magnetic bead capture with LC-MS/MS for therapeutic monoclonal antibody quantification in serum.

In recent years, there has been a growing interest in the thriving monoclonal antibody (mAb) industry due to the wide utilization of mAbs in clinical therapies. Robust and accurate bioanalytical methods are required to enable fast quantification of mAbs in biological matrices, especially in the context of pharmacokinetics (PKs)/pharmacodynamics (PDs) and therapeutic drug monitoring (TDM) studies. In this investigation, we presented a novel immuno-magnetic capture coupled with a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method designed for the quantification of immunoglobulin G-kappa-based mAbs in biological fluids. The immunoaffinity absorbent for mAb drug purification was meticulously crafted by immobilizing protein L onto monosize, magnetic poly(glycidyl methacrylate) (m-pGMA) beads, synthesized through dispersion polymerization. The microspheres were acquired with an average size of 1.6 μm, and the optimal binding of mAbs from the aqueous mAb solution was determined to be 45.82 mg g-1. The quantification of mAbs in 10 μL serum samples was achieved through affinity purification using m-pGMA@protein L beads (employing rituximab as an internal standard (IS)), on-bead reduction, and rapid tryptic digestion. Remarkably, the entire process, taking less than 2.5 hours, held significant potential for simplifying pretreatment procedures and minimizing analytical time. Furthermore, the developed method underwent validation in accordance with the European Medicines Agency (EMA) guidelines. The assay demonstrated commendable linearity within the 2-400 μg mL-1 range for both daratumumab and pembrolizumab. Intra- and inter-assay coefficients of variation fell within the range of 0.7% to 13.4%, meeting established acceptance criteria. Other validation parameters also conformed to regulatory standards. Ultimately, the efficacy of the method was substantiated in a pharmacokinetic study following a single-dose intravenous administration to mice, underscoring its applicability and reliability in real-world scenarios.

The myosin chaperone UNC-45 has an important role in maintaining the structure and function of muscle sarcomeres during adult aging.

C. elegans undergo age-dependent declines in muscle organization and function, similar to human sarcopenia. The chaperone UNC-45 is required to fold myosin heads after translation and is likely used for re-folding after thermally- or chemically-induced unfolding. UNC-45's TPR region binds Hsp90 and its UCS domain binds myosin heads. We observe early onset sarcopenia when UNC-45 is reduced at the beginning of adulthood. There is sequential decline of HSP-90, UNC-45, and MHC B myosin. A mutation in age-1 delays sarcopenia and loss of HSP-90, UNC-45 and myosin. UNC-45 undergoes age-dependent phosphorylation, and mass spectrometry reveals phosphorylation of 6 serines and 2 threonines, 7 of which occur in the UCS domain. Additional expression of UNC-45 results in maintenance of MHC B myosin and suppression of A-band disorganization in old animals. Our results suggest that increased expression or activity of UNC-45 might be a strategy for prevention or treatment of sarcopenia.

The Impact of Combined Sewer Overflows on Pharmaceutical and Illicit Drug Levels in New York/New Jersey Waterways.

Pharmaceuticals and drugs of abuse are organic micropollutants of emerging concern in both surface and groundwater worldwide. These compounds are considered to be pseudo-persistent because of their continuous release into water systems. The presence of these compounds in the environment at any concentration poses a potential risk to nontarget organisms. The main sources of these contaminants are wastewater treatment plants (WWTPs) and combined sewer overflows (CSOs). The primary goal of our study was to identify and quantify a panel of 28 commonly prescribed pharmaceuticals (mood-altering drugs, cardiovascular drugs, antacids, antibiotics) and high-prevalence drugs of abuse (cocaine, amphetamines, opioids, cannabis) in river water samples collected from 19 locations in the Hudson and East rivers in New York City. The second goal was to investigate the possible source (WWTP or CSOs) of these micropollutants. Samples were collected weekly from May to August 2021 (n = 224) and May to August 2022 (n = 232), and placed at -20 °C until analysis by liquid chromatography-tandem mass spectrometry. The most frequently detected analytes in 2021 were metoprolol (n = 206, 92%), benzoylecgonine (n = 151, 67%), atenolol (n = 142, 63%), and methamphetamine (n = 118, 53%), and in 2022 the most frequently detected were methamphetamine (n = 194, 84%), atenolol (n = 177, 76%), metoprolol (n = 177, 76%), and 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine (n = 159, 69%). Measured concentrations ranged from the limit of detection (0.50-5.00 ng/L) to 103 ng/L. More drugs and higher concentrations were detected in water contaminated by Enterococci (>60 most probably number) and after rainfall, indicating the influence of CSOs. The presence of drugs in samples with little to no Enterococci and after dry weather events indicates that WWTPs contribute to the presence of these substances in the river, probably due to a low removal rate. Environ Toxicol Chem 2024;00:1-12. © 2024 SETAC.