Search

Injecting CO 2 into deep geological formations can be an effective carbon removal and storage technology to mitigate global climate change. Interaction of injected CO 2 with rock formations changes pH and hydrochemistry within the deep injection zone (> 800 m depth). However, cap rocks and multiple tight aquitards typically act as barriers to protect the shallow aquifer from changes in the injection zone. Monitoring and evaluation of shallow groundwater quality are essential to verify that carbon capture storage projects (CCS) do not impact the near-surface environment. This study investigated shallow groundwater quality using long-term data (2006-2023) from a regular monitoring program at the Otway International Test Centre (OITC) in Victoria, Australia. It was found that shallow groundwater quality was stable over at least 15 years, during which time three phases of CO 2 injection into a deep storage zone occurred. The results highlighted groundwater quality complied with guidelines of Food and Agriculture Organization (FAO) and Australian water quality guidelines. Minor and localised changes observed in salinity or pH in shallow monitoring piezometers were caused by natural processes. Moreover, a wide range of groundwater quality indicators were evaluated. The results demonstrated that the groundwater quality of shallow aquifers (< 80 m) at OITC is suitable for agriculture. The study provides assurance and confidence to stakeholders that the quality of the near-surface environment has not been impacted by CO 2 injection into confined formations and no pollution has been detected. Although numerous CCS sites around the world are subject to monitoring, no evidence of changes in shallow groundwater quality has been reported that could be traced to CO 2 injection in confined formations at > 800 m depth., Competing Interests: Declarations. Ethical approval: The ethical approval requirement is not applicable. Consent to participate: All authors have significantly contributed to this work. Consent for publication: All authors agreed to publish. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Evening consumption of a whey protein rich in the amino acid tryptophan, alpha-lactalbumin (ALAC), has previously shown to benefit sleep-particularly among poor sleepers. Given trained populations often experience sleep difficulty, this study investigated whether evening supplementation of ALAC would influence sleep outcomes, mood, and next-day cognitive performance within a trained population with sleep difficulties. Nineteen trained participants (females, n = 11) with sleep difficulties (Athlete Sleep Screening Questionnaire: 8.1 ± 3.1; Pittsburgh Sleep Quality Index: 10.5 ± 4.1) completed this double-blinded, counterbalanced, randomized, crossover trial. Forty grams of ALAC or control were supplemented 2 hr presleep for three consecutive nights in a controlled environment, with sleep measured using dry electroencephalography. Blood samples were taken on the first evening of each experimental trial, with mood, sleepiness, and recovery assessed across the evening and morning. A cognitive testing battery was also completed each morning. During the ALAC condition, the primary findings were that participants had raised plasma tryptophan levels (p < .01), increased nonrapid eye movement Stage 2 sleep duration (CON: 205.9 ± 33.3; ALAC: 216.5 ± 33.1 min), reduced rapid eye movement duration (CON: 110.8 ± 27.9; ALAC: 99.7 ± 23.1 min), and improved reaction time in cognitive tests involving sensory motor speed, spatial orientation, and vigilant attention (p < .05). Data suggest evening supplementation of 40 g ALAC alters sleep architecture and improves next-morning reaction time in trained populations with sleep difficulties. Therefore, trained individuals experiencing sleep difficulty may benefit from acute ALAC supplementation to assist next-day performance. Future research should investigate this effect within habitual environments, outside of a tightly controlled setting.

Trace elements (TE) in living organisms can have detrimental health impacts depending on their concentration. As many TEs are obtained through diet, trophic niche changes associated with the impacts of anthropogenic activities and climate-change may influence exposure to top predators. The Australian fur seal (Arctocephalus pusillus doriferus; AUFS) represents the greatest resident, marine predator biomass in south-eastern Australia. With adult female foraging ranges limited to the continental shelf, their source of TEs is geographically restricted. Plasma, red blood cell and milk samples collected between 1998 and 2022 at Kanowna Island, were analysed for TEs (As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, Sn, V and Zn) using inductively-coupled plasma mass-spectrometry (ICP-MS). Plasma fatty-acid profiles and ocean climate variables were used to investigate trophic and environmental influences, respectively, on TE concentrations. Estimated whole blood concentrations in lactating females were comparable to levels reported in other marine mammals, except for Se and Mn. Correlations between adult tissues were negative for Mn and positive for As, Hg and Sn. Molar Se:Hg were high but within reported levels for pinnipeds. Element concentrations in pup plasma were greater than lactating females for Fe, Mn and Sn indicative of high transplacental transfer while doses of Se and As from milk exceeded tolerable effect levels for humans. Relationships with fatty-acid profiles suggest diet influenced concentrations of Cu, Hg, Mn, Sn, V and Zn in adult plasma. In addition, inter-annual variation in TE concentrations were influenced by broad-scale climate indices, including the Southern Annular Mode and the Southern Oscillation Index, and local conditions associated with the seasonally-active Bonney Upwelling. These findings indicate that TE concentrations in blood and milk of AUFS are and will continue to be affected by anticipated oceanographic changes, mediated by alterations in prey type availability, with potential impacts on the population's health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Improper waste disposal or inadequate wastewater treatment can result in pharmaceuticals reaching water bodies, posing environmental hazards. In this study, crude extracts containing the laccase enzyme from Pleurotus florida, Pleurotus eryngii, and Pleurotus sajor caju were used to degrade the fluoroquinolone antibiotics (FQs) levofloxacin (LEV), norfloxacin (NOR), ciprofloxacin (CIP), ofloxacin (OFL), and enrofloxacin (ENR) in aqueous solutions. The results for the fungi derived laccase extracts were compared with those obtained using commercially sourced laccase. Proteomics analysis of the crude extracts confirmed the presence of laccase enzyme across all three tested species, with proteins matching those found in Trametes versicolor and Pleurotus ostreatus. In vivo studies were conducted using species pure lines of fungal whole cells. The highest degradation efficiency observed was 77.7% for LEV in the presence of P. sajor caju after 25 days of treatment. Degradation efficiencies ranged from approximately 60-72% for P. florida, 45-76% for P. eryngii, and 47-78% for P. sajor caju. A series of in vitro experiments were also conducted using crude extracts from the three species and outcomes compared with those obtained when commercial laccase was used confirmed laccase as the enzyme responsible for antibiotic removal. The degradation efficiencies in vitro surpassed those measured in vivo, ranging from approximately 91-98% for commercial laccase, 77-92% for P. florida, 76-92% for P. eryngii, and 78-88% for P. sajor caju. Liquid chromatography-high-resolution mass spectrometry (LC-MS/MS) identified the degradation products, indicating a consistent enzymatic degradation pathway targeting the piperazine moiety common to all tested FQs, irrespective of the initial antibiotic structure. Phytoplankton toxicity studies with Dunaliella tertiolecta were performed to aid in understanding the impact of emerging contaminants on ecosystems, and by-products were analysed for ecotoxicity to assess treatment efficacy. Laccase-mediated enzymatic oxidation shows promising results in reducing algal toxicity, notably with Pleurotus eryngii extract achieving a 97.7% decrease for CIP and a 90% decrease for LEV. These findings suggest the potential of these naturally sourced extracts in mitigating antibiotic contamination in aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Non-degradable plastic mulch films used in agriculture are polluting the environment by leaving residues and microplastics in the soil. They are also difficult to recycle due to contamination during their use. Biodegradable mulch films are needed as alternatives so that they can be used effectively during the growing season and later be ploughed to be degraded in soil. However, market-available so-called biodegradable mulch films are very slow to degrade in the natural environment and thus do not fit with crop rotation demands or annual cultivation. In this study, we have developed mulch films from cotton gin trash (CGT) and/or gin motes (GM) in combination with biodegradable polycaprolactone and demonstrated their effectiveness over 3 months in outdoor conditions. Both the stability and degradation behaviours of mulch film samples were observed when they were placed on top of the soil and buried in the soil, respectively. Pesticide residue analysis also was carried out on CGT powder to identify and quantify individual pesticides against a matrix of known pesticides. The mulch films prepared in this study showed comparable and stable mechanical properties compared to commercial biodegradable mulch film, though were much quicker to degrade when buried in the soil. No pesticides were detected in the CGT samples. The films produced were vapour-permeable and may be useful in practical agricultural settings by being able to maintain consistent soil moisture and allowing precipitation to penetrate gradually. The lab-scale production cost for the film was 98.8 AUD/kg, which could be lowered by integrating a continuous film line in large-scale production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Ocular allergy (OA) is characterised by ocular surface itchiness, redness, and inflammation in response to allergen exposure. The primary aim of this study was to assess differences in the human tear metabolome and lipidome between OA and healthy controls (HCs) across peak allergy (spring-summer) and off-peak (autumn-winter) seasons in Victoria, Australia. A total of 19 participants (14 OA, 5 HCs) aged 18-45 were recruited and grouped by allergy questionnaire score. Metabolites and lipids from tear samples were analysed using mass spectrometry. Data were analysed using TraceFinder and Metaboanalyst. Metabolomics analysis showed 12 differentially expressed (DE) metabolites between those with OA and the HCs during the peak allergy season, and 24 DE metabolites were found in the off-peak season. The expression of niacinamide was upregulated in OA sufferers vs. HCs across both seasons ( p ≤ 0.05). A total of 6 DE lipids were DE between those with OA and the HCs during the peak season, and 24 were DE in the off-peak season. Dysregulated metabolites affected oxidative stress, inflammation, and homeostasis across seasons, suggesting a link between OA-associated itch and ocular surface damage via eye rubbing. Tear lipidome changes were minimal between but suggested tear film destabilisation and thinning. Such metabolipodome findings may pave new and exciting ways for effective diagnostics and therapeutics for OA sufferers in the future.

Background: Physiological adaptations during pregnancy alter nutrient and energy metabolism. Creatine may be important for maintaining cellular energy homeostasis throughout pregnancy. However, the impact of pregnancy on endogenous and exogenous creatine availability has never been comprehensively explored., Objectives: To undertake a prospective cohort study and determine the physiological ranges of creatine and associated metabolites throughout human pregnancy., Methods: Females with a singleton low-risk pregnancy were recruited at an Australian health service. Maternal blood and urine were collected at 5-time points from 10-36 weeks of gestation, and cord blood and placental samples were collected at birth. Creatine and associated amino acids and metabolites of creatine synthesis were analyzed. Dietary data were captured to determine effects of exogenous creatine intake. Associations between creatine metabolism and neonatal growth parameters were examined., Results: Two hundred and eighty-two females were included. Maternal plasma creatine remained stable throughout pregnancy [β: -0.003 μM; 95% confidence interval (CI): -0.07, 0.07; P = 0.94], though urinary creatine declined in late gestation (β: 0.38 μM/mmol/L creatinine (CRN); 95% CI: -0.47, -0.29; P < 0.0001). Plasma guanidinoacetate (GAA; the precursor to creatine during endogenous synthesis) fell from 10-29 weeks of gestation before rising until birth (β: -0.38 μM/mmol/L CRN; 95% CI: -0.47, -0.29; P < 0.0001). Urinary GAA followed an opposing pattern (β: 2.52 μM/mmol/L CRN; 95% CI: 1.47, 3.58, P < 0.001). Animal protein intake was positively correlated with maternal plasma creatine until ∼32 weeks of gestation (β: 0.07-0.18 μM; 95% CI: 0.006, 0.25; P ≤ 0.001). There were no links between creatine and neonatal growth, but increased urinary GAA in early pregnancy was associated with a slight reduction in head circumference at birth (β: -0.01 cm; 95% CI: -0.02, -0.004; P = 0.003)., Conclusions: Although maternal plasma creatine concentrations were highly conserved, creatine metabolism appears to adjust throughout pregnancy. An ability to maintain creatine concentrations through diet and shifts in endogenous synthesis may impact fetal growth. This trial was registered at [registry name] as ACTRN12618001558213., (Copyright © 2023 American Society for Nutrition. All rights reserved.)

Avian embryos develop in an egg composition which reflects both maternal condition and the recent environment of their mother. In birds, yolk corticosterone (CORT) influences development by impacting pre- and postnatal growth, as well as nestling stress responses and development. One possible mechanism through which maternal CORT may affect offspring development is via changes to offspring DNA methylation. We sought to investigate this, for the first time in birds, by quantifying the impact of manipulations to maternal CORT on offspring DNA methylation. We non-invasively manipulated plasma CORT concentrations of egg-laying female zebra finches (Taeniopygia castanotis) with an acute dose of CORT administered around the time of ovulation and collected their eggs. We then assessed DNA methylation in the resulting embryonic tissue and in their associated vitelline membrane blood vessels, during early development (5 days after lay), using two established methods - liquid chromatography-mass spectrometry (LC-MS) and methylation-sensitive amplification fragment length polymorphism (MS-AFLP). LC-MS analysis showed that global DNA methylation was lower in embryos from CORT-treated mothers, compared to control embryos. In contrast, blood vessel DNA from eggs from CORT-treated mothers showed global methylation increases, compared to control samples. There was a higher proportion of global DNA methylation in the embryonic DNA of second clutches, compared to first clutches. Locus-specific analyses using MS-AFLP did not reveal a treatment effect. Our results indicate that an acute elevation of maternal CORT around ovulation impacts DNA methylation patterns in their offspring. This could provide a mechanistic understanding of how a mother's experience can affect her offspring's phenotype., (© 2024 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Creatine metabolism likely contributes to energy homeostasis in the human uterus, but whether this organ synthesizes creatine and whether creatine metabolism is adjusted throughout the menstrual cycle and with pregnancy are largely unknown. This study determined endometrial protein expression of creatine-synthesizing enzymes arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), creatine kinase (CKBB), and the creatine transporter (SLC6A8) throughout the menstrual cycle in fertile and primary infertile women. It also characterized creatine metabolism at term pregnancy, measuring aspects of creatine metabolism in myometrial and decidual tissue. In endometrial samples, AGAT, GAMT, SLC6A8, and CKBB were expressed in glandular and luminal epithelial cells. Except for SLC6A8, the other proteins were also located in stromal cells. Irrespective of fertility, AGAT, GAMT, and SLC6A8 high-intensity immunohistochemical staining was greatest in the early secretory phase of the menstrual cycle. During the proliferative phase, staining for SLC6A8 protein was greater (P = 0.01) in the primary infertile compared with the fertile group. Both layers of the term pregnant uterus contained creatine, phosphocreatine, guanidinoacetic acid, arginine, glycine, and methionine; detectable gene and protein expression of AGAT, GAMT, CKBB, and ubiquitous mitochondrial CK (uMt-CK); and gene expression of SLC6A8. The proteins AGAT, GAMT, CKBB, and SLC6A8 were uniformly distributed in the myometrium and localized to the decidual glands. In conclusion, endometrial tissue has the capacity to produce creatine and its capacity is highest around the time of fertilization and implantation. Both layers of the term pregnant uterus also contained all the enzymatic machinery and substrates of creatine metabolism., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Seaweed extracts are a prominent class of biostimulants that enhance plant health and tolerance to biotic and abiotic stresses due to their unique bioactive components. However, the mechanisms of action of biostimulants are still unknown. Here, we have used a metabolomic approach, a UHPLC-MS method, to uncover the mechanisms induced following application to Arabidopsis thaliana of a seaweed extract derived from Durvillaea potatorum and Ascophyllum nodosum . We have identified, following the application of the extract, key metabolites and systemic responses in roots and leaves across 3 timepoints (0, 3, 5 days). Significant alterations in metabolite accumulation or reduction were found for those belonging to broad groups of compounds such as lipids, amino acids, and phytohormones; and secondary metabolites such as phenylpropanoids, glucosinolates, and organic acids. Strong accumulations of TCA cycle and N-containing and defensive metabolites such as glucosinolates were also found revealing the enhancement of carbon and nitrogen metabolism and defence systems. Our study has demonstrated that application of seaweed extract dramatically altered the metabolomic profiles of Arabidopsis and revealed differences in roots and leaves that varied across the timepoints tested. We also show clear evidence of systemic responses that were initiated in the roots and resulted in metabolic alterations in the leaves. Collectively, our results suggest that this seaweed extract promotes plant growth and activates defence systems by altering various physiological processes at the individual metabolite level., Competing Interests: Author TA is employed by Seasol International Pty Ltd and is an Adjunct Associate Professor at Deakin University. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Tran, Callahan, Islam, Wang, Arioli and Cahill.)

Wastewater treatment plants (WWTPs) are key components for the capture of microplastics (MPs) before they are released into natural waterways. Removal efficiencies as high as 99% may be achieved but sub-micron MPs as well as nanoplastics have been overlooked because of analytical limitations. Furthermore, short MP fibres are of concern because of their low capture rate as well as the lack of understanding of their influence on purification system efficiency. This study has investigated the impact of poly(ethylene terephthalate) (PET) short nanofibres on the performance of polyvinylidene fluoride (PVDF) ultrafiltration membranes during cross-flow operation. Model MP fibres with an average length of 10 ± 7 μm and a diameter of 142 ± 40 nm were prepared via a combination of electrospinning and fine cutting using a cryomicrotome. The manufactured MPs were added to both pure and synthetic domestic wastewater at a concentration of 1 mg.L -1 to determine their impact on the performance of PVDF ultrafiltration membranes. The results show that PET fibres attach to the membrane in a disorganised manner with low pore coverage. The water flux was decreased by 8% for MPs in pure water and no noticeable effect in wastewater after 3 days of filtration. Additionally, the nutrient removal efficiency of the membrane was not altered by the presence of PET MPs. These findings show that MP fibres do not significantly influence the early stages of filtration for a standard concentration of MPs in wastewater treatment plant studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Introduction: Chestnut rot caused by the fungus Gnomoniopsis smithogilvyi is a disease present in the world's major chestnut growing regions. The disease is considered a significant threat to the global production of nuts from the sweet chestnut (Castanea sativa). Conventional fungicides provide some control, but little is known about the potential of biological control agents (BCAs) as alternatives to manage the disease., Objective: Evaluate whether formulated BCAs and their secreted metabolites inhibit the in vitro growth of G. smithogilvyi., Methods: The antifungal potential of BCAs was assessed against the pathogen through an inverted plate assay for volatile compounds (VOCs), a diffusion assay for non-volatile compounds (nVOCs) and in dual culture. Methanolic extracts of nVOCs from the solid medium were further evaluated for their effect on conidia germination and were screened through an LC-MS-based approach for antifungal metabolites., Results: Isolates of Trichoderma spp., derived from the BCAs, significantly suppressed the pathogen through the production of VOCs and nVOCs. The BCA from which Bacillus subtilis was isolated was more effective in growth inhibition through the production of nVOCs. The LC-MS based metabolomics on the nVOCs derived from the BCAs showed the presence of several antifungal compounds., Conclusion: The results show that G. smithogilvyi can be effectively controlled by the BCAs tested and that their use may provide a more ecological alternative for managing chestnut rot. The in vitro analysis should now be expanded to the field to assess the effectiveness of these alternatives for chestnut rot management., (© 2022. The Author(s).)

Microalgae are important renewable feedstock to produce biodiesel and high-value chemicals. Different wavelengths of light influence the growth and metabolic activities of algae. Recent research has identified the light-sensing proteins called photoreceptors that respond to blue or red light. Structural elucidations of algal photoreceptors have gained momentum over recent years. These include channelrhodopsins, PHOT proteins, animal-like cryptochromes, and blue-light sensors utilizing flavin-adenine dinucleotide proteins. Pulsing light has also been investigated as a means to optimize energy inputs into bioreactors. This study summarizes the current structural and functional basis of photoreceptor modulation to optimize the growth, production of carotenoids and other high-value metabolites from microalgae. The review also encompasses novel photobioreactor designs that implement different light regimes including light wavelengths and time to optimize algal growth and desired metabolite profiles for high-value products., (© 2022 Wiley Periodicals LLC.)

Conventional breeding efforts for iron (Fe) and zinc (Zn) biofortification of bread wheat (Triticum aestivum L.) have been hindered by a lack of genetic variation for these traits and a negative correlation between grain Fe and Zn concentrations and yield. We have employed genetic engineering to constitutively express (CE) the rice (Oryza sativa) nicotianamine synthase 2 (OsNAS2) gene and upregulate biosynthesis of two metal chelators - nicotianamine (NA) and 2'-deoxymugineic acid (DMA) - in bread wheat, resulting in increased Fe and Zn concentrations in wholemeal and white flour. Here we describe multi-location confined field trial (CFT) evaluation of a low-copy transgenic CE-OsNAS2 wheat event (CE-1) over 3 years and demonstrate higher concentrations of NA, DMA, Fe, and Zn in CE-1 wholemeal flour, white flour, and white bread and higher Fe bioavailability in CE-1 white flour relative to a null segregant (NS) control. Multi-environment models of agronomic and grain nutrition traits revealed a negative correlation between grain yield and grain Fe, Zn, and total protein concentrations, yet no correlation between grain yield and grain NA and DMA concentrations. White flour Fe bioavailability was positively correlated with white flour NA concentration, suggesting that NA-chelated Fe should be targeted in wheat Fe biofortification efforts., (© 2021 Society for Experimental Biology and John Wiley & Sons Ltd.)

As academic staff scrambled into emergency remote teaching during COVID-19 restrictions, we also had to move in-person exams online without compromising integrity. This disruption caused us to think carefully about how chemistry is assessed, because 'business as usual' was no longer possible - and at some institutions, there are no plans for in-person exams to return., Competing Interests: Competing interestsThe authors declare no competing interests., (© Springer Nature Limited 2022.)

Antibiotic proliferation in the environment and their persistent nature is an issue of global concern as they induce antibiotic resistance threatening both human health and the ecosystem. Antibiotics have therefore been categorized as emerging pollutants. Fluoroquinolone (FQs) antibiotics are an emerging class of contaminants that are used extensively in human and veterinary medicine. The recalcitrant nature of fluoroquinolones has led to their presence in wastewater, effluents and water bodies. Even at a low concentration, FQs can stimulate antibacterial resistance. The main sources of FQ contamination include waste from pharmaceutical manufacturing industries, hospitals and households that ultimately reaches the wastewater treatment plants (WWTPs). The conventional WWTPs are unable to completely remove FQs due to their chemical stability. Therefore, the development and implementation of more efficient, economical, convenient treatment and removal technologies are needed to adequately address the issue. This review provides an overview of the technologies available for the removal of fluoroquinolone antibiotics from wastewater including adsorptive removal, advanced oxidation processes, removal using non-carbon based nanomaterials, microbial degradation and enzymatic degradation. Each treatment technology is discussed on its merits and limitations and a comparative view is presented on the choice of an advanced treatment process for future studies and implementation. A discussion on the commercialization potential and eco-friendliness of each technology is also included in the review. The importance of metabolite identification and their residual toxicity determination has been emphasized. The last section of the review provides an overview of the policy interventions and regulatory frameworks that aid in retrofitting antibiotics as a central key focus contaminant and thereby defining the discharge limits for antibiotics and establishing safe manufacturing practices., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Phosphorus (P) is an essential element for plant growth often limiting agroecosystems. To identify genetic determinants of performance under variable phosphate (Pi) supply, we conducted genome-wide association studies on five highly predictive Pi starvation response traits in 200 Arabidopsis (Arabidopsis thaliana) accessions. Pi concentration in Pi-limited organs had the strongest, and primary root length had the weakest genetic component. Of 70 trait-associated candidate genes, 17 responded to Pi withdrawal. The PHOSPHATE TRANSPORTER1 gene cluster on chromosome 5 comprises PHT1;1, PHT1;2, and PHT1;3 with known impact on P status. A second locus featured uncharacterized endomembrane-associated auxin efflux carrier encoding PIN-LIKES7 (PILS7) which was more strongly suppressed in Pi-limited roots of Pi-starvation sensitive accessions. In the Col-0 background, Pi uptake and organ growth were impaired in both Pi-limited pht1;1 and two pils7 T-DNA insertion mutants, while Pi -limited pht1;2 had higher biomass and pht1;3 was indistinguishable from wild-type. Copy number variation at the PHT1 locus with loss of the PHT1;3 gene and smaller scale deletions in PHT1;1 and PHT1;2 predicted to alter both protein structure and function suggest diversification of PHT1 is a key driver for adaptation to P limitation. Haplogroup analysis revealed a phosphorylation site in the protein encoded by the PILS7 allele from stress-sensitive accessions as well as additional auxin-responsive elements in the promoter of the "stress tolerant" allele. The former allele's inability to complement the pils7-1 mutant in the Col-0 background implies the presence of a kinase signaling loop controlling PILS7 activity in accessions from P-rich environments, while survival in P-poor environments requires fine-tuning of stress-responsive root auxin signaling., (© American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Agaricus xanthodermus and other species of the yellow-staining section Xanthodermatei are responsible for mushroom-related poisoning cases that require treatment. However, longstanding anecdotal evidence indicates that this species appears to exhibit considerable variation in toxicity, resulting in gastrointestinal irritation of varying severity in most cases. We quantified the amount of phenol, hydroquinone, and catechol in mushrooms using a novel protocol for gas chromatography-mass spectrometry (GC-MS) and investigated their levels in different basidiomatal structures, different developmental stages, and on different nutritional substrates. Phenol concentration was greater in the pileus than the stipe, in mature compared with immature basidiomata, and in basidiomata occurring in woody mulch versus lawns. Variation in toxicity is suggested to be due in part to variation in phenol concentration, developmental stage and tissue type consumed, and substrate. Variation in human sensitivity to simple phenols may also play a role but was not formally investigated in this study.

Dysregulation of histone deacetylases (HDACs) is associated with the pathogenesis of human osteosarcoma, which may present an epigenetic vulnerability as well as a therapeutic target. Domatinostat (4SC-202) is a next-generation class I HDAC inhibitor that is currently being used in clinical research for certain cancers, but its impact on human osteosarcoma has yet to be explored. In this study, we report that 4SC-202 inhibits osteosarcoma cell growth in vitro and in vivo. By analyzing cell function in vitro, we show that the anti-tumor effect of 4SC-202 involves the combined induction of cell-cycle arrest at the G2/M phase and apoptotic program, as well as a reduction in cell invasion and migration capabilities. We also found that 4SC-202 has little capacity to promote osteogenic differentiation. Remarkably, 4SC-202 revised the global transcriptome and induced distinct signatures of gene expression in vitro. Moreover, 4SC-202 decreased tumor growth of established human tumor xenografts in immunodeficient mice in vivo. We further reveal key targets regulated by 4SC-202 that contribute to tumor cell growth and survival, and canonical signaling pathways associated with progression and metastasis of osteosarcoma. Our study suggests that 4SC-202 may be exploited as a valuable drug to promote more effective treatment of patients with osteosarcoma and provide molecular insights into the mechanism of action of class I HDAC inhibitors.

Microplastics are ubiquitous in most biomes and environments, representing one of the most pressing global environmental challenges. This study investigated the ability of pre-production microplastic pellets to accumulate metals from the marine environment. An accidental ocean discharge of poly(propylene) pellets occurred via a wastewater treatment centre at the coastal city of Warrnambool, Victoria - Australia. These pellets were collected routinely from Shelly Beach, adjacent to the ocean discharge site over a period of 16-months following the spill. This collection formed a unique time-series that accurately represented the degree to which metal ions in the coastal marine environment accumulate on plastic debris. Elemental analysis indicated an increase in concentration over time of rare earth elements and a selection of other metals supporting the hypothesis that microplastics selectively adsorb metals from the environment. A subset of the poly(propylene) pellets contained a surfactant coating which significantly increased the adsorption capacity. The surface properties in relation to adsorption were further explored with surface imaging and these results are also discussed. This study shows how microplastics act as nucleation points and carriers of trace metal ions in marine environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: Humpback ( hpbk ) mice harbor a pathogenic mutation in the Notch3 gene and can serve as a beneficial animal model for investigating human myopathy, kyphosis, and developmental disorders, including lateral meningocele syndrome. Detection of the point mutation in hpbk mice is important for maintaining strains and scrutinizing genetic rescues, especially considering that homozygous mice are infertile and indistinguishable from their littermates at a young age. This study aimed for the development of a novel, precise, and time-saving genotyping method to identify the mutation in hpbk mice., Materials and Methods: In order to study the hpbk mouse line, we describe how we applied several tools, including quantitative polymerase chain reaction (qPCR), multiplex tetra-primer amplification-refractory mutation system (ARMS-PCR) and Sanger sequencing, toward the recognition of heterozygous and homozygous mice., Results: The Notch3 mutation was clearly identified using qPCR and ARMS assays, but the latter was a more precise and cost-effective approach. The lengths of the ARMS-PCR amplicons are 210 bp and 164 bp for the wild-type and hpbk alleles, respectively. Moreover, the genotyping results for each mouse were corroborated by Sanger DNA sequencing., Conclusion: Our newly developed PCR-based ARMS system affords a swift and precise way to genotype the hpbk mice. ARMS-PCR does not rely on any advanced equipment and is useful as a genotyping method for other model organisms that harbor a pathogenic variant., Competing Interests: The authors declare no financial conflict of interests., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Mixotrophic cultivation can increase microalgae productivity, yet the associated lipid metabolism remains mostly unknown. Stable isotope labeling was used to track assimilation of glycerol into the triacylglyceride (TAG) and membrane lipids of Nannochloropsis salina. In N-replete media, glycerol uptake and 13 C incorporation into acyl chains were, respectively, 6-fold and 12-fold higher than in N-deplete conditions. In N-replete cultures, 42% of the carbon in the consumed glycerol was assimilated into lipid acyl chains, mostly in membrane lipids rather than TAG. In N-deplete cultures, only 11% of the limited amount of consumed glycerol was fixed into lipid acyl chains. Labeled lipid-associated glycerol backbones were predominantly 13 C 3 labeled, suggesting that intact glycerol molecules were directly esterified with fatty acids/polar head groups. However, the presence of singly and doubly labeled lipid-bound glycerol species suggested that some glycerol also went through the central carbon metabolism before forming glycerol-3-phosphate destined for lipid esterification. 13 C incorporation was higher in the saturated and monounsaturated than the polyunsaturated acyl chains of TAG, indicating the flux of carbon from glycerol went first to de novo fatty acid synthesis before acyl editing reactions. The results demonstrate that nitrogen availability influences both glycerol consumption and utilization for lipid synthesis in Nannochloropsis, providing novel insights for developing mixotrophic cultivation strategies., (© 2020 Wiley Periodicals LLC.)

The analysis of 13 C-labeled lipids by mass spectrometry is challenging due to the complexity from labeling the large number of carbon atoms in lipids. To further add to the complexity, different adducts can be produced during electrospray ionization and in-source fragmentation, which can create complex overlapping isotope patterns that can only be resolved using high-resolution mass spectrometry. Co-elution of lipids even after chromatographic separation also adds to the potential for overlapping mass spectra. Here, we describe a procedure that enables full 13 C-labeled patterns to be resolved in complex microalgal lipid extracts as well a procedure that provides structural labeling information. Mass resolving powers of 240000 full width half-maximum (fwhm) and fast targeted MS/MS allowed the differentiation of isotopologues, adducts, and unresolved lipid species after chromatographic separation. This enabled the percentage of 13 C enrichment to be calculated for each individual lipid species over a time series in the microalgal lipidome. The application of tandem mass spectrometry (MS/MS) also allowed the degree of labeling within the headgroup vs acyl chains to be determined, further adding to the detail of information collected. This information is particularly useful for studying lipid synthesis and remodeling processes and can be extended to other biological systems.

Salinity-induced metabolic, ionic, and transcript modifications in plants have routinely been studied using whole plant tissues, which do not provide information on spatial tissue responses. The aim of this study was to assess the changes in the lipid profiles in a spatial manner and to quantify the changes in the elemental composition in roots of seedlings of four barley cultivars before and after a short-term salt stress. We used a combination of liquid chromatography-tandem mass spectrometry, inductively coupled plasma mass spectrometry, matrix-assisted laser desorption/ionization mass spectrometry imaging, and reverse transcription - quantitative real time polymerase chain reaction platforms to examine the molecular signatures of lipids, ions, and transcripts in three anatomically different seminal root tissues before and after salt stress. We found significant changes to the levels of major lipid classes including a decrease in the levels of lysoglycerophospholipids, ceramides, and hexosylceramides and an increase in the levels of glycerophospholipids, hydroxylated ceramides, and hexosylceramides. Our results revealed that modifications to lipid and transcript profiles in plant roots in response to a short-term salt stress may involve recycling of major lipid species, such as phosphatidylcholine, via resynthesis from glycerophosphocholine., (© 2019 John Wiley & Sons Ltd.)

Creatine is a metabolite important for cellular energy homeostasis as it provides spatio-temporal adenosine triphosphate (ATP) buffering for cells with fluctuating energy demands. Here, we examined whether placental creatine metabolism was altered in cases of early-onset pre-eclampsia (PE), a condition known to cause placental metabolic dysfunction. We studied third trimester human placentae collected between 27-40 weeks' gestation from women with early-onset PE ( n = 20) and gestation-matched normotensive control pregnancies ( n = 20). Placental total creatine and creatine precursor guanidinoacetate (GAA) content were measured. mRNA expression of the creatine synthesizing enzymes arginine:glycine aminotransferase ( GATM ) and guanidinoacetate methyltransferase ( GAMT ), the creatine transporter ( SLC6A8 ), and the creatine kinases (mitochondrial CKMT1A & cytosolic BBCK ) was assessed. Placental protein levels of arginine:glycine aminotransferase (AGAT), GAMT, CKMT1A and BBCK were also determined. Key findings; total creatine content of PE placentae was 38% higher than controls ( p < 0.01). mRNA expression of GATM (p < 0.001), GAMT (p < 0.001), SLC6A8 (p = 0.021) and BBCK (p < 0.001) was also elevated in PE placentae. No differences in GAA content, nor protein levels of AGAT, GAMT, BBCK or CKMT1A were observed between cohorts. Advancing gestation and birth weight were associated with a down-regulation in placental GATM mRNA expression, and a reduction in GAA content, in control placentae. These relationships were absent in PE cases. Our results suggest PE placentae may have an ongoing reliance on the creatine kinase circuit for maintenance of cellular energetics with increased total creatine content and transcriptional changes to creatine synthesizing enzymes and the creatine transporter. Understanding the functional consequences of these changes warrants further investigation., Competing Interests: The authors declare no conflict of interest.

Seed germination is the essential first step in crop establishment, and can be severely affected by salinity stress which can inhibit essential metabolic processes during the germination process. Salt stress during seed germination can trigger lipid-dependent signalling cascades that activate plant adaptation processes, lead to changes in membrane fluidity to help resist the stress, and cause secondary metabolite responses due to increased oxidative stress. In germinating barley ( Hordeum vulgare ), knowledge of the changes in spatial distribution of lipids and other small molecules at a cellular level in response to salt stress is limited. In this study, mass spectrometry imaging (MSI), liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS), inductively coupled plasma mass spectrometry (ICP-MS), and X-ray fluorescence (XRF) were used to determine the spatial distribution of metabolites, lipids and a range of elements, such as K + and Na + , in seeds of two barley genotypes with contrasting germination phenology (Australian barley varieties Mundah and Keel). We detected and tentatively identified more than 200 lipid species belonging to seven major lipid classes (fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, prenol lipids, sterol lipids, and polyketides) that differed in their spatial distribution based on genotype (Mundah or Keel), time post-imbibition (0 to 72 h), or treatment (control or salt). We found a tentative flavonoid was discriminant in post-imbibed Mundah embryos under saline conditions, and a delayed flavonoid response in Keel relative to Mundah. We further employed MSI-MS/MS and LC-QToF-MS/MS to explore the identity of the discriminant flavonoid and study the temporal pattern in five additional barley genotypes. ICP-MS was used to quantify the elemental composition of both Mundah and Keel seeds, showing a significant increase in Na + in salt treated samples. Spatial mapping of elements using µ-XRF localized the elements within the seeds. This study integrates data obtained from three mass spectrometry platforms together with µ-XRF to yield information on the localization of lipids, metabolites and elements improving our understanding of the germination process under salt stress at a molecular level., (Copyright © 2019 Gupta, Rupasinghe, Callahan, Natera, Smith, Hill, Roessner and Boughton.)