Glycolate Oxidase Research Articles

The Phytophthora infestans effector Pi05910 suppresses and destabilizes host glycolate oxidase StGOX4 to promote plant susceptibility.

Phytophthora infestans is a notorious oomycete pathogen that causes potato late blight. It secretes numerous effector proteins to manipulate host immunity. Understanding mechanisms underlying their host cell manipulation is crucial for developing disease resistance strategies. Here, we report that the conserved RXLR effector Pi05910 of P. infestans is a genotype-specific avirulence elicitor on potato variety Longshu 12 and contributes virulence by suppressing and destabilizing host glycolate oxidase StGOX4. By performing co-immunoprecipitation, yeast-two-hybrid assays, luciferase complementation imaging, bimolecular fluorescence complementation and isothermal titration calorimetry assays, we identified and confirmed potato StGOX4 as a target of Pi05910. Further analysis revealed that StGOX4 and its homologue NbGOX4 are positive immune regulators against P. infestans, as indicated by infection assays on potato and Nicotiana benthamiana overexpressing StGOX4 and TRV-NbGOX4 plants. StGOX4-mediated disease resistance involves enhanced reactive oxygen species accumulation and activated the salicylic acid signalling pathway. Pi05910 binding inhibited enzymatic activity and destabilized StGOX4. Furthermore, mutagenesis analyses indicated that the 25th residue (tyrosine, Y25) of StGOX4 mediates Pi05910 binding and is required for its immune function. Our results revealed that the core RXLR effector of P. infestans Pi05910 suppresses plant immunity by targeting StGOX4, which results in decreased enzymatic activity and protein accumulation, leading to enhanced plant susceptibility.

Zinc oxide nanoparticles at low dose mitigate lead toxicity in pea (Pisum sativum L.) seeds during germination by modulating metabolic and cellular defense systems

Addressing lead (Pb) pollution and contamination in soil and agriculture is urgent due to its widespread and long-lasting impacts on human health, food safety, and the environment. In the current study, we assessed the potential use of Zinc oxide nanoparticles (ZnONPs) in alleviating Pb toxicity in germinating seedlings. Pea (Pisum sativum L.) seedlings were exposed to Pb (83 mg/L) over a 7-day period, without and with ZnONPs amendment. Our results showed that the application of ZnONPs at 10 mg/L restored the morphological parameters affected by Pb, 34 %, 26 % and 23 %, for the length, fresh, and dry biomass of embryonic axis, respectively. ZnONPs decreased the activities of antioxidative enzymes of catalase (87.5 %), guaiacol peroxidase (60 %), and glutathione peroxidase (25 %), but increased the activities of ascorbate peroxidase (60 %) and glutathione reductase (25 %). These changes were associated with the increase (7 %) in the thiol groups, and reductions in the malondialdehyde (23 %), hydrogen peroxide (33 %), and carbonyl group levels (78 %), in addition to the stimulation of the activities of ROS-associated enzymes, including glycolate oxidase (40 %) and NADPH oxidase (by 89 %). Consequently, cell viability was increased by 66 %, while cellular death was reduced by 10 %, with ZnONPs, relative to the Pb-stressed seedlings. These findings highlight the mechanisms surrounding the ability of nanosized ZnO to mitigate the harmful effects of Pb on the growth and physiology of plants. Further investigation is needed to understand the mitigating effects of ZnONPs on Pb on the molecular and genomic levels in seedlings and plants, and ensure the sustainability of agricultural practices and food safety. Besides, site and source-specific integrated approaches must be practiced to formulate suitable remediation strategies.

Increased sugar content impairs pollen fertility and reduces seed-setting in high-photosynthetic-efficiency rice

Crop yield depends on biomass, which is primarily associated with photosynthesis. We previously demonstrated that two photorespiratory bypasses, i.e., GOC (glycolate oxidase + oxalate oxidase + catalase) and GCGT (glycolate oxidase + catalase + glyoxylate carboligase + tartronic semialdehyde reductase), significantly increased photosynthesis, biomass, and grain yield, but decreased seed-setting rates in rice. This study explored the underlying mechanism of how elevated photosynthetic efficiency impacted the seed-setting. First, pollen germination assessed in vivo and in vitro, revealed a reduced germination rate in GCGT rice. Subsequent analysis found that photosynthates highly accumulated in the leaves and stems; sucrose and soluble sugar levels were increased but the starch level was reduced in the anthers. Uridine diphosphate glucose (UDP-Glc) was increased but uridine diphosphate galactose (UDP-Gal) was unaltered, thus causing an imbalance in the UDP-Glc/UDP-Gal ratio in GCGT anthers. Most anthers in GCGT plants had two locules in contrast to four in the wild-type (WT). Pollen tapetum was developmentally abnormal, and genes related to sucrose synthesis, transport, and tapetal programmed cell death (PCD) were upregulated, whereas those involved in starch synthesis and conversion were downregulated in GCGT anthers. Taken together, our results demonstrated that an increase in sugar content was the primary factor causing reduced seed-setting rates in high photosynthetic efficiency rice, during which metabolic disorder of sugars and UDP sugar imbalance in anthers lead to impaired pollen fertility.

Metabolic engineering of Halomonas bluephagenesis for the production of ethylene glycol and glycolate from xylose

Halophilic Halomonas bluephagenesis, a natural producer of poly-3-hydroxybutyrate (PHB), was metabolically engineered to synthesize ethylene glycol and glycolate from xylose. Xylose utilization was achieved by overexpressing either the xylonate pathway or the ribulose-1-phosphate pathway. The key genes encoding for xylonate dehydratase and 2-keto-3-deoxy-xylonate aldolase in the xylonate pathway were screened. With further overexpressing aldehyde reductase gene yjgB, ethylene glycol accumulation was improved to 0.91g/L, accompanied with 1.48g/L of PHB accumulation. The disruption of native glycolate oxidase was found to be essential for glycolate production, and the defective recombinant strain produced 0.80g/L glycolate with 1.14g/L PHB in shake flask cultures. These results indicated that H. bluephagenesis has the potential to produce diverse metabolic chemicals from xylose.

Hydrogen Sulfide and 5‐Aminolevulinic Acid Synergistically Enhance Drought Tolerance in Tomato (Solanum lycopersicum L.)

ABSTRACTEnhancing crop drought tolerance is crucial for food security amid climate change. This study examines how 5‐aminolevulinic acid (ALA) and hydrogen sulfide (H2S) can improve drought resilience in tomato plants, which are essential for sustainable food production. Drought stress was induced using 12% PEG‐6000. Plants were pre‐treated with 25 mg L−1 ALA and 0.1 mg L−1 hypotaurine (HT), followed by 0.2 mM sodium hydrosulfide (NaHS) treatment to assess the effects on plant physiological effects over 10 days. Drought stress reduced plant dry weight, chlorophylls (a and b), Fv/Fm, leaf water potential, and relative water content, while increasing glycine betaine (GB) and proline levels. Additionally, drought stress elevated NADPH oxidase (NOX) and glycolate oxidase (GOX) activities, inducing oxidative stress and membrane damage. ALA and NaHS enhanced plant growth, photosynthesis, proline, GB, ALA content, ATP synthase, and ATPase activities, while mitigating NOX and GOX activities, thereby reducing and H2O2 radicals. ALA alone boosted L‐DES activity, promoting H2S accumulation. However, ALA + HT reduced H2S levels, compromising ALA's efficacy. NaHS with ALA + HT reinstated positive effects by restoring H2S levels. Biochemical assays confirmed ALA and NaHS promoted H2S accumulation, bolstering antioxidants, mitigating lipid peroxidation, suggesting their drought resilience potential in tomatoes.

Purification of Glycolate Oxidase Enzymes from Peasm (Pisum sativum l.) and Sorghum (Sorghum sudanense J.) Using Ion-Exchange Chromatography and their Physicochemical Characteristics

Purification of Glycolate Oxidase Enzymes from Peasm (Pisum sativum l.) and Sorghum (Sorghum sudanense J.) Using Ion-Exchange Chromatography and their Physicochemical Characteristics

Biochemical and gene expression profiling of oxalate biosynthesis in cherry tomato (Solanum lycopersicum var. cerasiforme) in Indo-Gangetic Plains of India

The oxalate content was assessed in nine diverse genotypes of cherry tomato in ripened fruit, leaf and root tissues. The maximum oxalate accumulation was found in leaf lamina followed by mature fruit and root tissues among all genotypes where the lowest oxalate content was found in the genotype FB-3-5 and the highest content was recorded in the genotype RSC-4. Examining the leaf and root of nine cherry tomato genotypes at three developmental stages, the investigation into the biosynthesis of oxalates focused on the study of two enzymes, glycolate oxidase (GLO) and ascorbate peroxidase (APX). A positive correlation was observed between oxalate formation and enzyme specific activity of both enzymes. Examining the relative gene expression of oxalate biosynthesizing genes, SlGLO2 and SlAPX7, indicated that the gene SlGLO2 encoded the GLO enzyme and played a crucial role in oxalate biosynthesis in cherry tomatoes. Understanding oxalate biosynthetic pathways and gene expression will aid in developing mechanisms to enhance cherry tomato nutritional quality and improving breeding programmes.

Effects of polystyrene microplastic composite with florfenicol on photosynthetic carbon assimilation of rice (Oryza sativa L.) seedlings: Light reactions, carbon reactions, and molecular metabolism

The effects of co-exposure to antibiotics and microplastics in agricultural systems are still unclear. This study investigated the effects of florfenicol (FF) and polystyrene microplastics (PS-MPs) on photosynthetic carbon assimilation in rice seedlings. Both FF and PS-MPs inhibited photosynthesis, while PS-MPs can alleviate the toxicity of FF. Chlorophyll synthesis genes (HEMA, HEMG, CHLD, CHLG, CHLM, and CAO) were down-regulated, whereas electron transport chain genes (PGR5, PGRL1A, PGRL1B, petH, and ndhH) were up-regulated. FF inhibited linear electron transfer (LET) and activated cyclic electron transfer (CET), which was consistent with the results of the chlorophyll fluorescence parameters. The photosynthetic carbon assimilation pathway was altered, the C3 pathway enzyme Ribulose1,5-bisphosphatecarboxylase/oxygenase (RuBisCO) was affected, C4 enzyme ((phosphoenolpyruvate carboxykinase (PEPCK), pyruvate orthophosphate dikinase (PPDK), malate dehydrogenase (MDH), and phosphoenolpyruvate carboxylase (PEPC))) and related genes were significantly up-regulated, suggesting that the C3 pathway is converted to C4 pathway for self-protection. The key enzymes involved in photorespiration, glycolate oxidase (GO) and catalase (CAT), responded positively, photosynthetic phosphorylation was inhibited, and ATP content and H+-ATPase activity were suppressed, nutrient content (K, P, N, Ca, Mg, Fe, Cu, Zn, Mn, and Ni) significantly affected. Transcriptomic analysis showed that FF and PS-MPs severely affected the photosynthetic capacity of rice seedlings, including photosystem I, photosystem II, non-photochemical quenching coefficients, and photosynthetic electron transport.

Luminescence-based complementation assay to assess target engagement and cell permeability of glycolate oxidase (HAO1) inhibitors

Glycolate oxidase (HAO1) catalyses the synthesis of glyoxylate, a common metabolic intermediate that causes renal failure if accumulated. HAO1 inhibition is an emerging treatment for primary hyperoxaluria, a rare disorder of glyoxylate metabolism. Here we report the first cell-based measurement of inhibitor uptake and engagement with HAO1, by adapting the cellular thermal shift assay (CETSA) based on Nano luciferase complementation and luminescence readout. By profiling the interaction between HAO1 and four well-characterised inhibitors in intact and lysed HEK293T cells, we showed that our CETSA method differentiates between low-permeability/high-engagement and high-permeability/low-engagement ligands and is able to rank HAO1 inhibitors in line with both recombinant protein methods and previously reported indirect cellular assays. Our methodology addresses the unmet need for a robust, sensitive, and scalable cellular assay to guide HAO1 inhibitor development and, in broader terms, can be rapidly adapted for other targets to simultaneously monitor compound affinity and cellular permeability.

In-Silico Enactment of Musa paradisiaca (L.) Exudate Against the Urotlithiatic Disease by Docking Study

Objectives: Urolithiasis is a condition that occurs when the stones exit the renal pelvis and move onto the remainder of the urinary collecting system. Musa paradisiaca (L.) is a plant with high nutritive value and has been used for the treatment of various diseases including urolithiasis. The objective of the present study was designed to detect the receptor-ligand binding energy and interaction through a molecular docking from the bio-active compounds in an exudates of M. paradisiaca pseudo stem on urolithiatic causative protein named as glycolate oxidase receptor (PDB ID: 2RDU). Methods: In silico study especially molecular docking was performed using AutodockVina and the best confirmation between ligand and protein were selected using Lamarkaina Genetic Algorithm (LGA) and ligand protein interaction was visualized using PyMol viewer. Novelty: Totally three various bio-active compounds are elucidated named as Olean-12-ene-3 beta, 28-diol, Tricyclo[8.4.1.1(3,8)] hexadeca- 3,5,7,10,12,14-hexaene- 2,9-dione, anti and 2H-Pyran,2-(7-heptadecynyloxy)tetrahydro- from the exudate of experimental sample. Three phyto active compounds or secondary metabolites were used for the present prediction by docking. The present result of molecular docking clearly revealed that olean 12 -ene-3 beta, 28 diol is the best biologically effective ligand observed through a highest docking score (-7.2 k cal/mol) on glycolate oxidase receptor than other two ligand compounds. In conclusion the compound olean 12 -ene-3 beta, 28 diol could be act as a potential lead molecule for urolithiasis. Keywords: Musa paradisiaca, Exudate, GCMS, Urolithiasis, AutoDock, PyMol Viewer

Multiple levels of transcriptional regulation control glycolate metabolism in Paracoccus denitrificans.

The hydroxyacid glycolate is a highly abundant carbon source in the environment. Glycolate is produced by unicellular photosynthetic organisms and excreted at petagram scales to the environment, where it serves as growth substrate for heterotrophic bacteria. In microbial metabolism, glycolate is first oxidized to glyoxylate by the enzyme glycolate oxidase. The recently described β-hydroxyaspartate cycle (BHAC) subsequently mediates the carbon-neutral assimilation of glyoxylate into central metabolism in ubiquitous Alpha- and Gammaproteobacteria. Although the reaction sequence of the BHAC was elucidated in Paracoccus denitrificans, little is known about the regulation of glycolate and glyoxylate assimilation in this relevant alphaproteobacterial model organism. Here, we show that regulation of glycolate metabolism in P. denitrificans is surprisingly complex, involving two regulators, the IclR-type transcription factor BhcR that acts as an activator for the BHAC gene cluster, and the GntR-type transcriptional regulator GlcR, a previously unidentified repressor that controls the production of glycolate oxidase. Furthermore, an additional layer of regulation is exerted at the global level, which involves the transcriptional regulator CceR that controls the switch between glycolysis and gluconeogenesis in P. denitrificans. Together, these regulators control glycolate metabolism in P. denitrificans, allowing the organism to assimilate glycolate together with other carbon substrates in a simultaneous fashion, rather than sequentially. Our results show that the metabolic network of Alphaproteobacteria shows a high degree of flexibility to react to the availability of multiple substrates in the environment.IMPORTANCEAlgae perform ca. 50% of the photosynthetic carbon dioxide fixation on our planet. In the process, they release the two-carbon molecule glycolate. Due to the abundance of algae, massive amounts of glycolate are released. Therefore, this molecule is available as a source of carbon for bacteria in the environment. Here, we describe the regulation of glycolate metabolism in the model organism Paracoccus denitrificans. This bacterium uses the recently characterized β-hydroxyaspartate cycle to assimilate glycolate in a carbon- and energy-efficient manner. We found that glycolate assimilation is dynamically controlled by three different transcriptional regulators: GlcR, BhcR, and CceR. This allows P. denitrificans to assimilate glycolate together with other carbon substrates in a simultaneous fashion. Overall, this flexible and multi-layered regulation of glycolate metabolism in P. denitrificans represents a resource-efficient strategy to make optimal use of this globally abundant molecule under fluctuating environmental conditions.

N-terminal acetylation orchestrates glycolate-mediated ROS homeostasis to promote rice thermoresponsive growth.

Climate warming poses a significant threat to global crop production and food security. However, our understanding of the molecular mechanisms governing thermoresponsive development in crops remains limited. Here we report that the auxiliary subunit of N-terminal acetyltransferase A (NatA) in rice OsNAA15 is a prerequisite for rice thermoresponsive growth. OsNAA15 produces two isoforms OsNAA15.1 and OsNAA15.2, via temperature-dependent alternative splicing. Among the two, OsNAA15.1 is more likely to form a stable and functional NatA complex with the potential catalytic subunit OsNAA10, leading to a thermoresponsive N-terminal acetylome. Intriguingly, while OsNAA15.1 promotes plant growth under elevated temperatures, OsNAA15.2 exhibits an inhibitory effect. We identified two glycolate oxidases (GLO1/5) as major substrates from the thermoresponsive acetylome. These enzymes are involved in hydrogen peroxide (H2O2) biosynthesis via glycolate oxidation. N-terminally acetylated GLO1/5 undergo their degradation through the ubiquitin-proteasome system. This leads to reduced reactive oxygen species (ROS) production, thereby promoting plant growth, particularly under high ambient temperatures. Conclusively, our findings highlight the pivotal role of N-terminal acetylation in orchestrating the glycolate-mediated ROS homeostasis to facilitate thermoresponsive growth in rice.

Heat shock combined with salinity impairs photosynthesis but stimulates antioxidant defense in rice plants

The interactive mechanisms that occur between heat shock and salt stress and their reflexes on photosynthesis and redox metabolism in plants are little understood yet. We tested the hypothesis that heat shock negatively interacts with salt stress, affecting most intensely K+/Na+ homeostasis, stomatal closure, and CO2 assimilation, in comparison to PSII activity and oxidative stress in rice leaves. 31-day old rice plants were previously exposed to 0 and 100 mM NaCl for eight days at 27 °C and afterwards two groups were transferred to high temperature (42 °C, heat shock) for 10 hours (heat and heat + salt) whereas two other groups remained at 27 °C (control and single salt treatments). Heat-salinity interaction greatly stimulated Na+ accumulation in leaves causing intense decrease in K+/Na+ ratios, inducing significant osmotic and ionic alterations. Stomata were closed intensely causing drastic impairment in CO2 assimilation and decrease in water use efficiency. In contrast, the PSII activity was much lesser affected, corroborated by a low increase in the fraction of closed reaction centers of PSII and slight decrease in electron transport rates. Despite that unbalance in photosynthesis, combined stress partially favored oxidative protection as indicated by a reduction in the levels of H2O2 and lipid peroxidation associated with reduction in the contents of reduced forms of ascorbate and glutathione. These favorable antioxidant responses were accompanied by increases in the activities of ascorbate peroxidases, superoxide dismutases, glutathione peroxidases, and phenol peroxidases whereas catalases and glycolate oxidases decreased. These antioxidant responses were not enough to mitigate overall physiological damages caused by combined stress, as indicated by drastic increase in membrane damage and decrease in relative water content in leaves. Heat shock drastically aggravates the negative effects caused by salt stress on the photosynthetic efficiency, especially CO2 assimilation, despite the heat – salinity interaction has partially favored the antioxidant defense.

Protective Role of Rosmarinic Acid in Experimental Urolithiasis: Understanding Its Impact on Renal Parameters.

This study aimed to assess the ability of rosmarinic acid (RA) to prevent kidney stone formation in an ethylene glycol and ammonium chloride (EG/AC) model. There was an increase in diuresis in the normotensive (NTRs) and hypertensive rats (SHRs) treated with hydrochlorothiazide (HCTZ) and exposed to EG/AC, while RA restored urine volume in NTRs. The EG/AC groups exhibited lower urine pH and electrolyte imbalance; these parameters were not affected by any of the treatments. Both HCTZ+EG/AC and RA+EG/AC reduced calcium oxalate crystal formation in NTR and SHR urine. Kidney tissue analysis revealed alterations in oxidative stress and inflammation parameters in all EG/AC-receiving groups, with RA enhancing antioxidant defenses in SHRs. Additionally, crystals were found in the kidney histology of all EG/AC-exposed groups, with reduced Bowman's capsule areas in NTRs and SHRs. The NTR VEH+EG/AC group showed intense renal damage, while the others maintained their structures, where treatments with HCTZ and RA were fundamental for kidney protection in the NTRs. Docking analysis showed that RA exhibited good binding affinity with matrix metalloproteinase-9, phosphoethanolamine cytidylyltransferase, and human glycolate oxidase enzymes. The data disclosed herein underscore the importance of further research to understand the underlying mechanisms better and validate the potential of RA for clinical use.

#190 Long-term treatment with lumasiran: final results from the phase 2 open-label extension study

Abstract Background and Aims Primary hyperoxaluria type 1 (PH1) is a rare genetic disorder in which hepatic oxalate overproduction can lead to kidney failure and life-threatening systemic disease. Lumasiran, an RNA interference (RNAi) therapeutic indicated for treatment of PH1 to lower urinary oxalate (UOx) and plasma oxalate (POx) levels, reduces hepatic oxalate production by degrading the mRNA encoding glycolate oxidase. Patients with PH1 who completed a Phase 1/2 study were eligible to enroll in a Phase 2 open-label extension (OLE) study (NCT03350451) within 12 months of completing the Phase 1/2 study. Method Patients enrolled in the 54-month Phase 2 OLE had previously met eligibility criteria for the Phase 1/2 study, including: age 6 to 64 years, PH1 confirmed by genetic criteria, 24-hour (24 h) UOx excretion >0.7 mmol/1.73 m2/day, estimated glomerular filtration rate (eGFR) >45 mL/min/1.73 m2, and a stable pyridoxine (vitamin B6) regimen for patients taking vitamin B6. In the Phase 2 OLE, patients were to initiate subcutaneous lumasiran 1.0 mg/kg once monthly (qM), 3.0 mg/kg qM, or 3.0 mg/kg every 3 months (q3M) based on their dose/regimen in the Phase 1/2 study. The primary endpoint was the incidence of adverse events (AEs). Secondary endpoints were change in 24 h UOx corrected for body surface area (BSA) over time, change in 24 h UOx:creatinine ratio (UOx:Cr) over time, and change in estimated glomerular filtration rate (eGFR) over time. Change over time in POx and plasma glycolate, incidence of anti-drug antibodies (ADAs), and rates of kidney stone AEs were also assessed. Results All 20 patients (median age 11.5 years; 65% female) who completed the Phase 1/2 study entered the Phase 2 OLE (Table 1). Most patients (65%) were taking vitamin B6 at baseline. All patients transitioned to the 3.0 mg/kg q3M dose/regimen by month (M) 21, and all patients completed the study. Six patients transitioned to commercially available lumasiran prior to M54 but on or after M45. The most common treatment-related AEs were injection-site reactions (ISRs) with 13 events in 8 patients (40%); all ISRs were graded mild. Of all lumasiran doses administered during the OLE, 3% (13 of 427 total doses of lumasiran) were associated with ISRs. Other than ISRs, the only treatment-related AEs affecting >1 patient were increases in bilirubin (N=3 patients; 15%); all events resolved without lumasiran dosing changes. Severe AEs were reported by 2 patients (10%); serious AEs were reported by 7 patients (35%). No deaths occurred. No severe or serious AEs were related to treatment, and no patients discontinued lumasiran or withdrew due to AEs. Lumasiran treatment led to a substantial reduction in 24 h UOx relative to the Phase 1/2 study-derived baseline (Fig. 1A). At M54, the mean absolute change from baseline in 24 h UOx was −1.5 mmol/24 h/1.73 m2 and the mean percent change was −68%. From M42 through M54, 24 h UOx was ≤1.5×ULN in ≥89% of patients. At M54, the mean percent change from baseline in 24 h UOx:Cr was −77%. Mean eGFR was generally stable over time (Fig. 1B). Mean absolute change from baseline in eGFR at M54 was −1.6 mL/min/1.73 m2. Lumasiran treatment also led to reductions in POx from baseline, and mean post-baseline POx values remained within the normal range through M54. Mean plasma glycolate levels were elevated at the start of the Phase 2 OLE due to persistence of the treatment effect from the Phase 1/2 study. After treatment resumed, levels increased and then plateaued. One patient (5%) had an ADA positive result at M6 but no ADA-related AEs; subsequent tests were negative through M48. Overall, the kidney stone AE rate was low during the Phase 2 OLE (0.17/person-year (PY]); by comparison, the rate of symptomatic kidney stones was 0.45/PY during the 12 months prior to the Phase 1/2 study and the kidney stone AE rate was 0.90/PY during the Phase 1/2 study. Conclusion In the longest follow-up reported to date, lumasiran treatment was well tolerated in patients with PH1 in the Phase 2 OLE. The most common treatment-related AEs were mild, transient ISRs; no severe or serious AEs were treatment related. Mean 24 h UOx reductions were sustained over more than 4.5 years of treatment. Mean eGFR was stable over time.

Acclimation of intertidal macroalgae Ulva prolifera to UVB radiation: the important role of alternative oxidase

BackgroundSolar radiation is primarily composed of ultraviolet radiation (UVR, 200 − 400 nm) and photosynthetically active radiation (PAR, 400 − 700 nm). Ultraviolet-B (UVB) radiation accounts for only a small proportion of sunlight, and it is the primary cause of plant photodamage. The use of chlorofluorocarbons (CFCs) as refrigerants caused serious ozone depletion in the 1980s, and this had led to an increase in UVB. Although CFC emissions have significantly decreased in recent years, UVB radiation still remains at a high intensity. UVB radiation increase is an important factor that influences plant physiological processes. Ulva prolifera, a type of macroalga found in the intertidal zone, is intermittently exposed to UVB. Alternative oxidase (AOX) plays an important role in plants under stresses. This research examines the changes in AOX activity and the relationships among AOX, photosynthesis, and reactive oxygen species (ROS) homeostasis in U. prolifera under changes in UVB and photosynthetically active radiation (PAR).ResultsUVB was the main component of solar radiation impacting the typical intertidal green macroalgae U. prolifera. AOX was found to be important during the process of photosynthesis optimization of U. prolifera due to a synergistic effect with non-photochemical quenching (NPQ) under UVB radiation. AOX and glycolate oxidase (GO) worked together to achieve NADPH homeostasis to achieve photosynthesis optimization under changes in PAR + UVB. The synergism of AOX with superoxide dismutase (SOD) and catalase (CAT) was important during the process of ROS homeostasis under PAR + UVB.ConclusionsAOX plays an important role in the process of photosynthesis optimization and ROS homeostasis in U. prolifera under UVB radiation. This study provides further insights into the response of intertidal macroalgae to solar light changes.

Potassium and jasmonic acid —Induced nitrogen and sulfur metabolisms improve resilience against arsenate toxicity in tomato seedlings

Despite the well-known individual roles of potassium (K+) and jasmonic acid (JA) in plant responses to various stresses, their combined effects on improving tolerance to metalloid arsenate (AsV) toxicity and their influence on physiological and biochemical mechanisms remain largely unexplored. Therefore, the present study was conducted to explore the concomitant role of K+ and JA in nitrogen (N) and sulfur (S) metabolisms and regulation of antioxidant system in tomato seedlings under AsV toxicity conditions. The obtained data reveal that K+ and/or JA boosted the tolerance of tomato (Solanum lycopersicum L.) seedlings to AsV toxicity. However, the effect of a combined dose of K+ and JA was found to be more efficient as compared to the alone application. Seedlings subjected to AsV exhibited suppressed morphological attributes (shoot and root length, shoot and root fresh weight, and shoot and root dry weight), and physio-biochemical characteristics. Also, AsV-treated tomato seedlings showed enhanced reactive oxygen species (ROS) and increased the activity of ROS-producing enzymes (O2•— -generating NADPH oxidase activity and H2O2-generating glycolate oxidase] and also a chlorophyll (Chl) degrading enzyme [chlorophyllase (Chlase) activity]. However, the exogenous supply of K+ and JA synergistically inhibited Chl degradation and overproduction of ROS by suppressing their responsible enzymes activity. Moreover, it induced gas exchange parameters and activity of enzymes responsible for the photosynthesis process (carbonic anhydrase and Rubisco), Chl biosynthesis (δ-aminolevulinic acid dehydratase), and suppressed Chlase activity, and overproduction of ROS and their producing enzymes activity. Most interestingly, the combined application of K+ and JA substantially increased N and S content and their assimilation by upregulating the activity of enzymes involved in these key pathways (nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, ATP sulfurylase, adenosine 5-phosphosulfate reductase, O-acetylserine (thiol) lyase and sulfide reductase). The obtained results strongly reveal that the combined application of K+ and JA improved tolerance of tomato seedlings to AsV toxicity by enhancing N and S assimilation pathways and antioxidant system.

Characterization of OsPIN2 Mutants Reveal Novel Roles for Reactive Oxygen Species in Modulating Not Only Root Gravitropism but Also Hypoxia Tolerance in Rice Seedlings.

Tolerance to submergence-induced hypoxia is an important agronomic trait especially for crops in lowland and flooding-affected areas. Although rice (Oryza sativa) is considered a flood-tolerant crop, only limited cultivars display strong tolerance to prolonged submergence and/or hypoxic stress. Therefore, characterization of hypoxic resistant genes and/or germplasms have important theoretical and practical significance for rice breeding and sustained improvements. Previous investigations have demonstrated that loss-of-function of OsPIN2, a gene encoding an auxin efflux transporter, results in the loss of root gravitropism due to disrupted auxin transport in the root tip. In this study, we revealed a novel connection between OsPIN2 and reactive oxygen species (ROS) in modulating root gravitropism and hypoxia tolerance in rice. It is shown that the OsPIN2 mutant had decreased accumulation of ROS in root tip, due to the downregulation of glycolate oxidase encoding gene OsGOX6, one of the main H2O2 sources. The morphological defects of root including waved rooting and agravitropism in OsPIN2 mutant may be rescued partly by exogenous application of H2O2. The OsPIN2 mutant exhibited increased resistance to ROS toxicity in roots due to treatment with H2O2. Furthermore, it is shown that the OsPIN2 mutant had increased tolerance to hypoxic stress accompanied by lower ROS accumulation in roots, because the hypoxia stress led to over production of ROS in the roots of the wild type but not in that of OsPIN2 mutant. Accordingly, the anoxic resistance-related gene SUB1B showed differential expression in the root of the WT and OsPIN2 mutant in response to hypoxic conditions. Notably, compared with the wild type, the OsPIN2 mutant displayed a different pattern of auxin distribution in the root under hypoxia stress. It was shown that hypoxia stress caused a significant increase in auxin distribution in the root tip of the WT but not in that of the war1 mutant. In summary, these results suggested that OsPIN2 may play a role in regulating ROS accumulation probably via mediating auxin transport and distribution in the root tip, affecting root gravitropism and hypoxic tolerance in rice seedlings. These findings may contribute to the genetic improvement and identification of potential hypoxic tolerant lines in rice.

Lumasiran: A Review in Primary Hyperoxaluria Type 1.

Lumasiran (Oxlumo®), a first-in-class synthetic, double-stranded, ribonucleic acid (RNA) interference molecule targeting glycolate oxidase through silencing HAO1 mRNA, is approved in several countries for patients of any age and stage of kidney function with primary hyperoxaluria type 1 (PH1). Approval was based on results from the phase III ILLUMINATE trials. In the double-blind, placebo-controlled, ILLUMINATE-A trial, subcutaneous lumasiran was significantly more effective than placebo in reducing 24-h urinary oxalate excretion in patients aged ≥6 years with PH1; this effect was sustained for ≥36 months in ongoing longer-term analyses. In the single-arm ILLUMINATE-B trial, lumasiran reduced urinary oxalate:creatinine ratios and plasma oxalate levels in patients aged <6 years with PH1. In the single-arm ILLUMINATE-C trial, lumasiran reduced plasma oxalate levels in patients with PH1 receiving dialysis as well as those not receiving dialysis. In secondary and exploratory analyses of these trials, nephrocalcinosis grade, kidney stone event rates and estimated glomerular filtration rates were either stable or improved with lumasiran. Lumasiran had an acceptable tolerability profile that remained consistent in longer-term analyses; the most common adverse events were mild and transient injection-site reactions. Thus, lumasiran is an effective treatment option, with an acceptable tolerability profile, in patients with PH1.

Efficient and safe therapeutic use of paired Cas9-nickases for primary hyperoxaluria type 1

The therapeutic use of adeno-associated viral vector (AAV)-mediated gene disruption using CRISPR-Cas9 is limited by potential off-target modifications and the risk of uncontrolled integration of vector genomes into CRISPR-mediated double-strand breaks. To address these concerns, we explored the use of AAV-delivered paired Staphylococcus aureus nickases (D10ASaCas9) to target the Hao1 gene for the treatment of primary hyperoxaluria type 1 (PH1). Our study demonstrated effective Hao1 gene disruption, a significant decrease in glycolate oxidase expression, and a therapeutic effect in PH1 mice. The assessment of undesired genetic modifications through CIRCLE-seq and CAST-Seq analyses revealed neither off-target activity nor chromosomal translocations. Importantly, the use of paired-D10ASaCas9 resulted in a significant reduction in AAV integration at the target site compared to SaCas9 nuclease. In addition, our study highlights the limitations of current analytical tools in characterizing modifications introduced by paired D10ASaCas9, necessitating the development of a custom pipeline for more accurate characterization. These results describe a positive advance towards a safe and effective potential long-term treatment for PH1 patients.