Absolute Amounts Research Articles

Absolute quantitation of binding antibodies from clinical samples

The quantitation of antibody responses is a critical requirement for the successful development of vaccines and therapeutics that often relies on the use of standardized reference materials to determine relative quantities within biological samples. The validity of comparing responses across assays using arbitrarily defined reference values is therefore limited. We developed a generalizable method known as MASCALE (Mass Spectrometry Enabled Conversion to Absolute Levels of ELISA Antibodies) for absolute quantitation of antibodies by calibrating ELISA reference sera using mass spectrometry. Levels of proteotypic peptides served as a proxy for human IgG, allowing the conversion of responses from arbitrary values to absolute amounts. Applications include comparison of binding assays at two separate laboratories and evaluation of cross-clade magnitude-breadth responses induced by an investigational HIV-1 vaccine regimen. MASCALE addresses current challenges in the interpretation of immune responses in clinical trials and expands current options available to make suitable comparisons across different settings.

Quantification of heritable variation in multiple lodgepole pine chemical and physical traits that contribute to defense against mountain pine beetle (Dendroctonus ponderosae)

Bark beetles are major causes of conifer mortality worldwide. We quantified genetic contributions to key chemical and physical traits in lodgepole pine that contribute to defense against mountain pine beetle, a native tree-killing herbivore. Assays were conducted with mature trees in replicated provenance trials. Among chemical defenses, monoterpene composition (relative proportions of various compounds) had higher genetic variability than monoterpene concentration (absolute quantities). Additionally, constitutive concentrations had higher genetic variabilities than induced concentrations elicited by simulated attack. Resin flow elicited by mechanical wounding likewise showed very low genetic variability. In previous studies with this and related systems, induced monoterpene concentration and resin flow are the two best predictors respectively of which trees will survive outbreaks under natural conditions. This means that expression of the most efficacious defense traits is strongly influenced by environmental factors. These low heritabilities despite their survival values likely arise from specific life history and ecological aspects of this system, such as the lengthy period between when lodgepole pines begin cone production and become large enough for brood development, the extended viability of serotinous cones post-mortem, the high heterogeneity of both resources and stressors that trees experience across landscapes, and a strong proximate-neighbor effect on tree mortality arising from beetle aggregation pheromones. However, low heritability values do not preclude long-term evolvability and coadaptation of defenses, for which there is extensive mechanistic and associational evidence in this system. Among other traits we evaluated, sesquiterpenes had higher genetic variability than monoterpenes. Sesquiterpenes are not known to influence conifer-bark beetle interactions but have anti-herbivore properties in some other plant-insect systems. The lengths of lesions formed in response to simulated attack showed no heritable component and likewise do not predict tree survival in nature. Bark texture was highly heritable and is known to influence beetle post-landing orientation and colonization patterns, but whether it influences survival patterns at the whole-tree level is unknown. Relationships between tree defenses and growth were generally positive, suggesting that environmental heterogeneity in resource quality among trees supersedes potential underlying substrate allocation processes within trees. Similarly, relationships between various defense traits were generally positive rather than showing trade-offs, suggesting conifer resistance to native bark beetles entails coordinated defense syndromes. Implications to tree breeding, forest management, and coevolution between long-lived plants and rapidly reproducing herbivores are discussed. The extent to which these trends may apply to other bark beetle systems is evaluated, along with specific priorities for future research.

Household debt and children’s psychological well-being in China: The mediating role of parent–child relations

BackgroundHousehold debt is a psychological stressor for those who take the debt on, but it remains unclear whether and how this money-related stressor influences children’s psychological well-being. AimsThe study investigated the association of household debt with children’s psychological well-being and further explored the mediating role of parent–child relations. MethodsData were obtained from two waves (2016 and 2018) of the China Family Panel Studies, with a sample of 1,828 children included. The Center for Epidemiological Studies of Depression Scale was applied to measure psychological well-being. Household debt was assessed by absolute and relative amounts of total debt, mortgage debt, and non-mortgage debt. We used positive and negative parent–child interaction to measure parent–child relations. The characteristics of children, parents, and households were included as covariates. ResultsHousehold debt, especially mortgage debt, was positively associated with psychological illnesses among children, and the link was substantially explained by parent–child relations. ConclusionHousehold debt exerted detrimental effects on children’s psychological well-being by jeopardizing parent–child relations in China. Strategies that address the need of vulnerable children are required to reduce the potentially negative psychological well-being outcomes of household debt.

Multi-speciation and ignition delay time measurements of ammonia oxidation behind reflected shock waves

Ammonia (NH3) oxidation was studied using emission and laser absorption diagnostics in a shock tube. Experiments were carried out using stoichiometric mixtures of NH3 and O2 highly dilute in Ar seeded with CO2 at post-reflected-shock conditions from 1700–2500K near 2atm. Five optical diagnostics in the infrared (IR), visible, and ultraviolet (UV) were implemented to measure NH3 (before and after shock-heating), NO, electronically excited amino radicals (NH2∗), electronically excited hydroxyl radicals (OH∗), and temperature during oxidation. To mitigate mixture uncertainty due to NH3 adsorption, a scanned-wavelength laser absorption diagnostic near 10.4μm was developed to measure initial NH3 concentration. A two-color, fixed-wavelength laser absorption diagnostic near 225 nm was developed to measure quantitative NO and NH3 time-histories. A detailed spectroscopic model of the NO A2Σ+←X2Π system was used to provide NO absorption cross-sections at both wavelengths, and NH3 absorption cross-sections were measured at the conditions of interest. Emission from NH2∗ near 600 nm was collected to provide qualitative insight into radical formation prior to ignition, and emission from OH∗ near 308 nm was used to determine ignition delay times (IDTs). A two-color, fixed-wavelength laser absorption diagnostic at 4.17μm and 4.19μm was developed to measure temperature using small quantities of seeded CO2. Measured time-histories are compared against predictions from two kinetic models. Experimental results and model predictions show several discrepancies, both in terms of timescales and absolute quantities. Sensitivity analysis shows that the reactions H + O2⇌ O + OH, NH3 + M ⇌ NH2 + H + M, and NH3 + H ⇌ NH2 + H2 are dominant across all conditions, and that NH3 pyrolysis reactions in general drive much of the chemistry. Direct, low-scatter rate constant measurements of the NH3 + M ⇌ NH2 + H + M reaction and additional multi-speciation measurements of NH3 pyrolysis and oxidation at combustion-relevant temperatures and pressures are recommended.Novelty and significance statementThe novelty of this work is exemplified most strongly by the use of UV laser absorption diagnostics to measure NH3 and NO during high-temperature oxidation of NH3. Simultaneous measurements of temperature via CO2 laser absorption at two wavelengths and light emission from NH2∗ and OH∗ are also novel. Simultaneous, time-resolved, quantitative measurements of NH3, NO, and temperature and qualitative measurements of NH2∗ and OH∗ light emission are of significant utility as a model validation-suited data set. Additionally, the data presented herein and the associated findings provide evidence of current chemical kinetic model deficiencies pertaining to NH3 chemistry and suggest avenues for model improvement.

A Novel Tissue-Specific Insight into Sex Steroid Fluctuations Throughout the Murine Estrous Cycle.

Serum sex steroid levels fluctuate throughout the reproductive cycle. However, the degree to which sex steroid tissue content mimics circulating content is unknown. Understanding the flux and physiological quantity of tissue steroid content is imperative for targeted hormonal therapy development. Utilizing a gold-standard ultrasensitive liquid chromatography-mass spectrometry (LC/MS) method we determined sex steroid (17β-estradiol [E2], testosterone, androstenedione, and progesterone) fluctuations in serum and in 15 tissues throughout the murine estrous cycle (proestrus, estrus, and diestrus I) and in ovariectomized (OVX) mice. We observed dynamic fluctuations in serum and tissue steroid content throughout the estrous cycle with proestrus generally presenting the highest content of E2, testosterone, and androstenedione, and lowest content of progesterone. In general, the trend in circulating steroid content between the stages of the estrous cycle was mimicked in tissue. However, the absolute amounts of steroid levels when normalized to tissue weight were found to be significantly different between the tissues with the serum steroid quantity often being significantly lower than the tissue quantity. Additionally, we found that OVX mice generally displayed a depletion of all steroids in the various tissues assessed, except in the adrenal glands which were determined to be the main site of peripheral E2 production after ovary removal. This investigation provides a comprehensive analysis of steroid content throughout the estrous cycle in a multitude of tissues and serum. We believe this information will help serve as the basis for the development of physiologically relevant, tissue-specific hormonal therapies.

Eye proteome of Drosophila melanogaster.

Drosophila melanogaster is a popular model organism to elucidate the molecular mechanisms that underlie the structure and function of the eye as well as the causes of retinopathies, aging, light-induced damage, or dietary deficiencies. Large-scale screens have isolated genes whose mutation causes morphological and functional ocular defects, which led to the discovery of key components of the phototransduction cascade. However, the proteome of the Drosophila eye is poorly characterized. Here, we used GeLC-MS/MS to quantify 3516 proteins, including the absolute (molar) quantities of 43 proteins in the eye of adult male Drosophila reared on standard laboratory food. This work provides a generic and expandable resource for further genetic, pharmacological, and dietary studies.

Natural variation of temperature acclimation of Arabidopsis thaliana.

Acclimation is a multigenic trait by which plants adjust photosynthesis and metabolism to cope with a changing environment. Here, natural variations of photosynthetic efficiency and acclimation of the central carbohydrate metabolism were analyzed in response to low and elevated temperatures. For this, 18 natural accessions of Arabidopsis thaliana, originating from Cape Verde Islands and Europe, were grown at 22°C before being exposed to 4°C and 34°C for cold and heat acclimation, respectively. Absolute amounts of carbohydrates were quantified together with their subcellular distribution across plastids, cytosol and vacuole. Linear electron transport rates (ETRs) were determined together with the maximum quantum efficiency of photosystem II (Fv/Fm) for all growth conditions and under temperature fluctuation. Under elevated temperature, ETR residuals under increasing photosynthetic photon flux densities significantly correlated with the degree of temperature fluctuation at the original habitat of accessions, indicating a geographical east/west gradient of photosynthetic acclimation capacities. Plastidial sucrose concentrations positively correlated with maximal ETRs under fluctuating temperature, indicating a stabilizing role within the chloroplast. Our findings revealed specific subcellular carbohydrate distributions that contribute differentially to the photosynthetic efficiency of natural Arabidopsis thaliana accessions across a longitudinal gradient. This sheds light on the relevance of subcellular metabolic regulation for photosynthetic performance in a fluctuating environment and supports the physiological interpretation of naturally occurring genetic variation of temperature tolerance and acclimation.

Change of histone H3 lysine 14 acetylation stoichiometry in human monocyte derived macrophages as determined by MS-based absolute targeted quantitative proteomic approach: HIV infection and methamphetamine exposure

BackgroundHistones posttranslational modification represent an epigenetic mechanism that regulate gene expression and other cellular processes. Quantitative mass spectrometry used for the absolute quantification of such modifications provides further insight into cellular responses to extracellular insults such as infections or toxins. Methamphetamine (Meth), a drug of abuse, is affecting the overall function of the immune system. In this report, we developed, validated and applied a targeted, MS-based quantification assay to measure changes in histone H3 lysine 14 acetylation (H3K14Ac) during exposure of human primary macrophages to HIV-1 infection and/or Meth.MethodsThe quantification assay was developed and validated to determine H3K14Ac stoichiometry in histones that were isolated from the nuclei of control (CIC) and exposed to Meth before (CIM) or/and after (MIM) HIV-infection human monocyte-derived macrophages (hMDM) of six donors. It was based on LC–MS/MS measurement using multiple reaction monitoring (MRM) acquisition of the unmodified and acetylated form of lysine K14 of histone H3 9KSTGGKAPR17 peptides and the corresponding stable isotope labeled (SIL) heavy peptide standards of the same sequences. The histone samples were propionylated (Poy) pre- and post- trypsin digestion so that the sequences of the monitored peptides were: K[Poy]STGGK[1Ac]APR, K[Poy]STGGK[1Ac]APR-heavy, K[Poy]STGGK[Poy]APR and K[Poy]STGGK[Poy]APR-heavy. The absolute amounts of the acetylated and unmodified peptides were determined by comparing to the abundances of their SIL standards, that were added to the samples in the known concentrations, and, then used for calculation of H3K14Ac stoichiometry in CIC, CIM and MIM hMDM.ResultsThe assay was characterized by LLOD of 0.106 fmol/µL and 0.204 fmol/µL for unmodified and acetylated H3 9KSTGGKAPR17 peptides, respectively. The LLOQ was 0.5 fmol/µL and the linear range of the assay was from 0.5 to 2500 fmol/µL. The absolute abundances of the quantified peptides varied between the donors and conditions, and so did the H3K14Ac stoichiometry. This was rather attributed to the samples nature itself, as the variability of their triplicate measurements was low.ConclusionsThe developed LC–MS/MS assay enabled absolute quantification of H3K14Ac in exposed to Meth HIV-infected hMDM. It can be further applied determination of this PTM stoichiometry in other studies on human primary macrophages.Graphical

OR33-03 Circadian Analysis Of Nuclear Receptor Protein Expression In Mouse Liver Reveals Peak COUP-TFII, EAR2, LXRa, LXRb And TR4 Expression At The End Of The Mouse Active Cycle ZT0

Abstract Disclosure: M.F. Saikali: None. J. Li: None. C.L. Cummins: None. Nuclear Receptors (NRs) are a family of ligand-mediated transcription factors that control the expression of genes involved in a wide range of physiological processes. The ability to study NRs quantitatively at the protein level has been hindered by the poor quality of available antibodies, as well as the lack of available high throughput methods to study more than a few receptors at a time. To address this need, we have developed a mass spectrometry based targeted proteomic assay to quantify the absolute amounts of NR protein in mouse livers over a circadian period. C57Bl6 mice were sacrificed at 12 weeks every 4 hours from zeitgeber time ZT0 to ZT20. Their livers were processed using a transcription factor extraction protocol1. For each sample, 100 µg nuclear protein was processed using the NR assay described previously2. Briefly, the lysate is digested following the Multi-Enzyme Digestion Filter Aided Sample Preparation protocol. The sample is then spiked with 50 fmol of our internal standard mix. This mix is composed of 66 carefully selected peptides that account for 49 NRs, 11 histone peptides, and 6 circadian clock proteins. The resulting digest that contains the internal standards is desalted on C18 tips, followed by separation on an EASY-Spray C18 column (75 um x 50 cm, 3Å), and analyzed on a Thermo QExactive HF mass spectrometer in parallel reaction monitoring mode. We used this assay to quantify the changes in NR protein expression in mouse livers for 9 receptors; RORα, CAR, COUP-TFII, EAR2, GR, LXRα, LXRβ, PPARα, and TR4. RORα showed peak nuclear expression at ZT0, with a peak-to-trough change of 2.2-fold. COUP-TFII, EAR2, LXRα, LXRβ, and TR4 also showed peak expression at ZT0 (the end of the active phase) with changes of 3.3, 2.4, 1.7, 2.0, and 1.6-fold respectively. GR showed peak nuclear expression at ZT12 with a 1.8-fold change. CAR showed 2.3-fold peak expression at ZT16. COUP-TFII, EAR2, and TR4 are considered orphan receptors with no known endogenous ligands. These data will be used to uncover endogenous tissue-specific ligands by analyzing extracts isolated at the zeitgeber time that the receptor is maximally expressed. Sources of Research Support: This project has been supported by the J. P. Bickell Foundation and the Canadian Institutes of Health Research New Frontiers Research Fund and International Development Research Fund. 1.Gillespie, M. A. et al. Absolute Quantification of Transcription Factors Reveals Principles of Gene Regulation in Erythropoiesis. Mol Cell (2020) doi:10.1016/j.molcel.2020.03.031.2.Saikali, M. F. & Cummins, C. L. Quantifying the Protein Levels of All Nuclear Hormone Receptors by Mass Spectrometry. J Endocr Soc5, (2021). Presentation: Sunday, June 18, 2023

Evaluation of amino acids and other related metabolites levels in end-stage renal disease (ESRD) patients on hemodialysis by LC/MS/MS and GC/MS.

End-stage renal disease (ESRD) is a rapidly increasing health problem, and every year, about 2 million ESRD cases are reported worldwide. Hemodialysis (HD) is the vital renal reinstatement therapy for ESRD, and HD patterns play a crucial role in patients' health. Plasma metabolomics is the potential approach to understanding the HD process, effectiveness, and patterns. The lack of protein vitality is a primary problem for HD patients, and the quantities of amino acids intracellularly and in theblood are considered to be a symbolic index of protein metabolism and nutrition conditions. In the current study, LC/MS/MS and GC/MS methods were developed for 29 targeted plasma metabolites and validated as per ICH bioanalytical method validation M10 guidelines. The 29 metabolites included 20 proteinogenic amino acids and nine other related metabolites. The methods were employed to measure the absolute quantities (µM) of the targeted metabolites in HD patients (n=60) before and after dialysis (PRE-HD and POST-HD), and compared with the healthy control (HC) group (n=60). Phenylacetylglutamine was found to be higher in both PRE-HD (72.88±14.5 µM) and POST-HD (26.62±7.9 µM), when compared to HC (1.61±0.6 µM). On the other hand, glutamic acid was lower in PRE-HD (14.90±6.5 µM), and POST-HD (13.6±6.1 µM) than that of HC (245.4±37.8 µM). The dialytic loss was found to be 52-45% for arginine, lysine, and histidine, while it was 38-26% for glycine, cysteine, proline, alanine, threonine, glutamine, valine, and methionine. The dialytic loss was low (≤12%) for aspartic acid, glutamic acid, asparagine, leucine, tyrosine, tryptophan, and isoleucine. Graphical abstract adapted from mass spectrometry templates by Biorender.com retrieved from https://app.biorender.com/biorender-templates .

Low Concentrations of Antibiotics Alter Microbial Communities and Induce High Abundances of Antibiotic-Resistant Genes in Ornamental Water

Antibiotic resistance poses a significant threat to the public health domain. A favorable platform for generating and disseminating antibiotic-resistant genes (ARGs) and antibiotic-resistant bacteria (ARB) is provided by landscaped fish ponds created by urbanization. This research delved into the effects exerted by different concentrations of specific antibiotics, namely tetracycline and ciprofloxacin, on the microbial community composition present in water samples obtained from a landscape pond. Additionally, we analyzed the abundance of ARGs and the class 1 integron-integrase gene (intI1), and identified potential hosts of ARGs. The results indicated that the consistent administration of antibiotics significantly influenced the microbial community structure, resulting in variations within both bacterial communities and functionalities. Furthermore, the absolute quantities of ARGs, including tetA, tetC, qnrA, and qnrS, as well as the integrase gene intI1, exhibited augmentation in response to varying types and concentrations of antibiotics. Notably, the regular input of low concentrations of antibiotics produced higher levels of abundance of ARGs than the regular input of higher concentrations of antibiotics. The use of different types of antibiotics led to diverse host bacteria structures.

Content of myeloid-derived suppressor cells in autoimmune diseases in children

Myeloid-derived suppressor cells (MDSCs) play an important role in regulation of immune response. An increase in their number in adult patients with autoimmune diseases has been reported. G-MDSCs, M-MDSCs, and MDSCs(M-G-) at different stages of autoimmune disease may both activate T cell proliferation, leading to disease progression, or inhibit it, thus promoting Treg differentiation. Arginase-1 (Arg- 1) is an enzyme in MDSCs that reduces the concentration of arginine required for T lymphocyte proliferation. Our aim was to evaluate the content of MDSCs populations and functional activity of MDSCs in children with autoimmune diseases. 75 children with inflammatory bowel diseases (IBD), 60 children with multiple sclerosis (MS), 69 children with psoriasis (PS), 62 healthy age-matched children were included into the study. The content of MDSCs ((CD3, CD19, CD56, HLA-DR)-, CD11b+ and CD33+), subpopulations of MDSCs (M-MDSCs, G-MDSCs expressing CD14 and CD15), assessment of Arg-1 activity were performed by flow cytometry techniques. The content of MDSCs in patients with IBD, MS and PS was significantly higher than in the comparison group and depended on the state of exacerbation/remission. In exacerbation and remission of IBD, MS and PS, a significant increase of MDSCs was revealed when compared with healthy children; the highest values were found in children in exacerbation of MS (Me-3.5 (2.5-5.6) % MNC against Me-1.6 (0.9-2.5) % MNC, p 0.001). In patients with MS, the content of G-MDSC, M-MDSC was significantly higher, and MDSC(M-G-) was lower than in healthy children. An increase in absolute amounts of G-MDSCs was shown in MS exacerbation compared to the disease remission state (p = 0.022). For patients with IBD, a significant increase in percentage of MDSCs and M-MDSCs (p = 0.014 and p = 0.045, respectively) was obtained in exacerbation of the disease relative to remission state. In patients with IBD, MS, and PS, a significant increase in Arg-1 activity in MDSCs was found, with a decreased number of MDSCs in patients in remission compared to exacerbation phase of the disease. In children with autoimmune diseases, an increase in the MDSC populations was found. The activity of arginase-1 in MDSCs is increased in remission, along with a decrease in their numbers.

Regional difference in precipitation seasonality over China from CMIP6 projections

AbstractBased on historical simulations and projections under two scenarios of the Shared Socioeconomic Pathways (SSP2‐4.5 and SSP5‐8.5) provided by 32 models from the Coupled Model Intercomparison Project Phase 6 (CMIP6), we investigate the future precipitation seasonality changes over China by two indices, the dimensionless seasonality index (DSI) and the relative seasonality index (SI), and the associated physical mechanisms by moisture budget diagnosis. The results show that relative to the baseline period 1995–2014, the DSI is enhanced across almost all of China at the end of the 21st century, with stronger magnitudes in southeastern China and the southern Tibetan Plateau and weaker magnitudes in northwestern China, and national‐mean enhancements are 15% (7%–21%) and 23% (15%–29%) for the multimodel median (25th–75th percentile range across models) under the SSP2‐4.5 and SSP5‐8.5 scenarios, respectively. Projected SI changes show large regional differences, with decreases in northern China and increase in southern China, leading to little change for the national mean. The DSI and SI changes are associated with uneven distributions of changes in absolute amounts and relative proportions of precipitation among seasons, respectively. Further analysis reveals that the contributions of different moisture budget terms to precipitation changes vary across seasons, resulting in changes in the seasonal distribution of precipitation and thus the DSI and SI changes. Specifically, the thermodynamic component of vertical moisture advection primarily determines the greater magnitude of increased precipitation in summer than in winter in most of China, resulting in the DSI change. The reduction in residual term dominates the diminished winter precipitation in southern China, leading to more precipitation concentrated in summer for given annual precipitation and thus the intensification in SI, while the opposite holds true in northern China.

A quantitative sequencing method using synthetic internal standards including functional and phylogenetic marker genes.

The method of spiking synthetic internal standard genes (ISGs) to samples for amplicon sequencing, generating sequences and converting absolute gene numbers from read counts has been used only for phylogenetic markers and has not been applied to functional markers. In this study, we developed ISGs, including gene sequences of the 16S rRNA, pmoA, encoding a subunit of particulate methane monooxygenase and amoA, encoding a subunit of ammonia monooxygenase. We added ISGs to the samples, amplified the target genes and performed amplicon sequencing. For the mock community, the copy numbers converted from read counts using ISGs were equivalent to those obtained by the quantitative real-time polymerase chain reaction (4.0 × 104 versus 4.1 × 104 and 3.0 × 103 versus 4.0 × 103 copies μL-DNA-1 for 16S rRNA and pmoA genes, respectively), but we also identified underestimation, possibly due to primer coverage (7.8 × 102 versus 3.7 × 103 μL-DNA-1 for amoA gene). We then applied this method to environmental samples and analysed phylogeny, functional diversity and absolute quantities. One Methylocystis population was most abundant in the sludge samples [16S rRNA gene (3.8 × 109 copies g-1 ) and the pmoA gene (2.3 × 109 copies g-1 )] and were potentially interrelated. This study demonstrates that ISG spiking is useful for evaluating sequencing data processing and quantifying functional markers.

Data implementation matters: Effect of software choice and LCI database evolution on a comparative LCA study of permanent magnets

AbstractLife cycle assessment (LCA) databases and software evolve. We analyzed to which extent software and evolving life cycle inventory databases affect the comparison of technology alternatives, using a comparative LCA on permanent magnets as a case study, with two selected software tools: CMLCA and Brightway LCA. We migrated the system models from the CMLCA to Brightway LCA software and alternated between the ecoinvent database versions 2.2 and 3.1 to 3.6 in the system background. When using ecoinvent v3.6 instead of v2.2, the change of the indicator results ranged from to 283%. The evolution of the ecoinvent database impacted the absolute amounts of the characterized results and the relative performance between alternatives. The impact category with the highest variability was ionizing radiation, which even showed a ranking inversion with ecoinvent v3.4. In contrast, the impact of using CMLCA or Brightway was negligible because the same data and modeling assumptions caused percentage differences below 0.4%. During the semi‐automated data migration to Brightway, we identified 23 environmental flows in the CMLCA model that were not paired with their corresponding characterization factors in the published study of reference. This error had led to an underestimation of 63% in the photochemical oxidation indicator of one of the alternatives. This underestimation relates to an interoperability issue regarding the nomenclature of environmental flows in software alternatives and is a matter of data implementation rather than an issue intrinsic to the selected software. Finally, we identified improvement opportunities for the transparency and reusability of LCA models. This article met the requirements for a Gold‐Gold JIE data openness badge described at http://jie.click/badges.

Lignocellulose Degradation Efficiency of Agaricus bisporus Strains Grown on Wheat Straw-Based Compost.

The cultivation of Agaricus bisporus was investigated with two commercial strains, A15 and W192. Nitrogen and lignocellulose were analyzed in absolute amounts based on mass balance to accurately compare the degradation efficiency of the compost, and the correlation between the degradation efficiency and extracellular enzyme activity of the mycelium was analyzed. Lignocellulose utilization efficiency positively correlated with mushroom yield. For the same strain, the compost with high lignocellulose content resulted in high utilization efficiency, which increased the yield of A. bisporus. For the same compost, the lignocellulose utilization efficiency of A15 was higher than that of W192. The activities of manganese-dependent peroxidase and β-glucosidase indicated that W192 may have a higher demand for lignin and cellulose. Therefore, a higher yield of W192 was obtained with high-lignocellulose compost. The metabolism of cellulose and hemicellulose in the mycelial growth stage seemed to be conducive to high mushroom yield.

Triple-Isotope Tracing for Pathway Discernment of NMN-Induced NAD+ Biosynthesis in Whole Mice.

Numerous efforts in basic and clinical studies have explored the potential anti-aging and health-promoting effects of NAD+-boosting compounds such as nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN). Despite these extensive efforts, our understanding and characterization of their whole-body pharmacodynamics, impact on NAD+ tissue distribution, and mechanism of action in various tissues remain incomplete. In this study, we administered NMN via intraperitoneal injection or oral gavage and conducted a rigorous evaluation of NMN's pharmacodynamic effects on whole-body NAD+ homeostasis in mice. To provide more confident insights into NMN metabolism and NAD+ biosynthesis across different tissues and organs, we employed a novel approach using triple-isotopically labeled [18O-phosphoryl-18O-carbonyl-13C-1-ribosyl] NMN. Our results provide a more comprehensive characterization of the NMN impact on NAD+ concentrations and absolute amounts in various tissues and the whole body. We also demonstrate that mice primarily rely on the nicotinamide and NR salvage pathways to generate NAD+ from NMN, while the uptake of intact NMN plays a minimal role. Overall, the tissue-specific pharmacodynamic effects of NMN administration through different routes offer novel insights into whole-body NAD+ homeostasis, laying a crucial foundation for the development of NMN as a therapeutic supplement in humans.

Service sweethearting: An effective way to increase tips?

Survey research suggests that a common tactic servers use to increase their tips is to comp (or fail to charge for) a portion of customers’ food orders. An online hypothetical scenario study found that this tactic does result in larger absolute tip amounts. The effect was fully mediated by the comp’s enhancement of the tipper’s feelings of happiness, luckiness, and gratitude. These positive affective reactions to the comp enhanced tips even though the comp lowered the bill size upon which consumers based their tip amounts. These findings suggest that restaurant managers cannot rely upon consumer perceptions of unexpected comps as unethical or as attempts at manipulation to keep them from rewarding such employee theft. Nor can managers rely upon the percentage tipping norm together with the lower bill sizes created by such give-aways to provide disincentives for this form of employee theft.

WRF-Chem quantification of transport events and emissions sensitivity in Korea during KORUS-AQ

To quantify the relative roles of long-range transport (LRT) versus locally emitted aerosol and ozone precursors during polluted periods in Korea, high-resolution (4 km) Weather Research and Forecasting with Chemistry model simulations were performed. The model was evaluated using surface and airborne observations collected during the KORea and United States Air Quality campaign. Ozone above 40 ppb had mean bias of −5.9 ppb. PM2.5 was biased high (8.2 µg/m3), with a relative bias of 30% given the mean observed value of 26.8 µg/m3. The absolute amounts and shifts between phases for all PM2.5 species except nitrate reasonably match observations across all 4 phases. Notable limitations include an underestimation of nighttime planetary boundary layer height. Transport versus domestic emissions influence was studied by model runs with perturbed emissions and by comparing east-west fluxes over the Yellow Sea to Korean emissions and other normalization metrics. Domestic anthropogenic emission contributions to surface air quality were quantified by location across Korea, segregated by synoptic meteorological phase. The largest contributions from Korean emissions were found under high-pressure stagnant conditions and the smallest for conditions with strong westerly winds. For example, at Seoul, domestic contributions of PM2.5 averaged 49% and 29% in the aforementioned meteorological phases, respectively. Surface concentrations of NOx and toluene in Seoul were over 85% due to domestic emissions. CO and black carbon had both local and remote contributions. Nitrate and ammonium contributions varied greatly by phases in Seoul, with 7%–51% nitrate and 42%–70% of ammonium from remote sources. Variation in direction (west-to-east vs. east-to-west) and magnitude of fluxes support the model sensitivity results. Analysis using fluxes facilitates the quantification of source contributions for secondary species and, in many cases, can be done using a single model run or reanalysis result. The analysis presented shows the importance of using models with high spatial resolution to capture pollutant transport and mixing around Korea. However, there remain uncertainties in secondary aerosol production mechanisms and indications that local production at times could be higher than those modeled in this analysis. Therefore, the results presented here should be viewed as an upper limit on the importance of LRT.

A Shifted Urinary Microbiota Associated with Disease Activity and Immune Responses in Rheumatoid Arthritis.

Recent evidence emphasized the role of the microbiota in the etiopathogenesis of rheumatoid arthritis (RA). Indeed, it has been demonstrated that urinary tract infections are implicated in RA pathogenesis. However, a definitive association between the urinary tract microbiota and RA remains to be investigated. Urine samples from 39 patients affected by RA, including treatment-naive patients, and 37 age- and sex-matched healthy individuals were collected. In RA patients, the urinary microbiota showed an increase in microbial richness and a decrease in microbial dissimilarity, especially in treatment-naive patients. A total of 48 altered genera with different absolute quantities were detected in patients with RA. The 37 enriched genera included Proteus, Faecalibacterium, and Bacteroides, while the 11 deficient genera included Gardnerella, Ruminococcus, Megasphaera, and Ureaplasma. Notably, the more abundant genera in RA patients were correlated with the disease activity score of 28 joints-erythrocyte sedimentation rates (DAS28-ESR) and an increase in plasma B cells. Furthermore, the altered urinary metabolites, such as proline, citric acid, and oxalic acid, were positively associated with RA patients, and they were closely correlated with urinary microbiota. These findings suggested a strong association between the altered urinary microbiota and metabolites with disease severity and dysregulated immune responses in RA patients. IMPORTANCE We revealed that the profile of the urinary tract microbiota in RA featured with increased microbial richness and shifted taxa, associated with immunological and metabolic changes of the disease, underlining the interplay between urinary microbiota and host autoimmunity.