Pyruvate Kinase Activity Research Articles

Influence of Dietary Phosphorus on the Growth, Feed Utilization, Proximate Composition, Intestinal Enzymes, and Oxidation Resistance of Sea Cucumber Apostichopus japonicus.

Six experimental diets (crude protein 12.58%, crude fat 1.93%, and total energy 10.72 kJ/kg) containing 0.24%, 0.37%, 0.51%, 0.62%, 0.77%, and 0.89% phosphorus were formulated to evaluate dietary phosphorus requirement for sea cucumber Apostichopus japonicus. The feeding trial was conducted in 18 fiberglass tanks (220 L) for 63 days. Each diet was randomly assigned to triplicate tanks of 50 sea cucumbers (9.99 g) and fed once daily. With the increase of dietary phosphorus level, weight gain (WG), specific growth rate (SGR), daily feed intake (DFI), feces production ratio, the activities of amylase, alkaline phosphatase, phosphofructokinase, succinate dehydrogenase, and glutathione peroxidase as well as the contents of glutathione and glutathione oxidized significantly increased and then decreased afterwards (P < 0.05). A. japonicus fed diet with 0.63%, 0.63%, and 0.55% dietary phosphorus was estimated to yield the highest WG (11.39 g), SGR (1.09%/d), and DFI (2.55%/d) according to the quadratic regression analysis of WG, SGR, and DFI against dietary phosphorus level, respectively. The apparent digestibility of dry material and energy followed an opposite tendency. Feed efficiency, the contents of whole-body phosphorus, initially increased and then plateaued, fitting piecewise-linear models with breakpoint at 0.57% and 0.55% dietary phosphorus. Daily phosphorus intake, pyruvate kinase activity, and the ratio of glutathione and glutathione oxidized increased (P < 0.05) but the apparent digestibility of phosphorus, the activities of alkaline protease, aspartate transaminase, and phosphoenolpyruvate carboxykinase decreased (P < 0.05), responding to the increasing dietary phosphorus. Considering the present results, the optimal dietary phosphorus for A. japonicus is 0.57-0.63%.

Exogenous nitric oxide inhibits the respiratory metabolism of postharvest wax apple fruit and its role in the delayed cottony softening

This study investigated the impacts of nitric oxide (NO) fumigation on the respiratory metabolism of postharvest wax apple fruit and its impact on fruit cottony softening. NO treatment inhibited the respiration rate of wax apple fruit and delayed the development of cottony softening; NO treatment suppressed the activities of hexokinase, phosphohexose isomerase (PGI), pyruvate dehydrogenase, and succinate dehydrogenase (SDH) in fruit pulp, but improved the activities of pyruvate kinase and citrate synthase (CS), and maintained the contents of glucose and pyruvic acid, indicating a reduction in the EMP-TCA pathway. Moreover, the NADH content and the activities of SDH and cytochrome C oxidase (CCO) were decreased with NO treatment, while the NADH/NAD ratio increased, suggesting that the electron transport chain complexes of wax apple fruit were reduced. Besides, NO treatment enhanced the activity of NADK in wax apple fruit, which lowered the levels of NAD and NADH but increased levels of NADP and NADPH, indicating an increase in pentose phosphate pathway. Additionally, CATPCA and heatmap analysis show that the PGI, CS, SDH, and CCO played a crucial role in NO-inhibited respiration. Collectively, these findings indicate that NO treatment suppressed the respiratory metabolism of postharvest wax apple fruit via regulating respiratory pathways, which reduces fruit senescence and delays cottony softening.

Oral pyruvate prevents high-intensity interval exercise-induced metabolic acidosis in rats by promoting lactate dehydrogenase reaction.

There is no denying the clinical benefits of exogenous pyruvate in the treatment of pathological metabolic acidosis. However, whether it can prevent exercise physiological metabolic acidosis, delay the occurrence of exercise fatigue, and improve the beneficial effects of exercise and its internal mechanism remain unclear. We randomly divided 24 male SD rats into 3 groups: one group was a control without exercise (CC, n = 8), and the other two groups were supplemented with 616 mg/kg/day pyruvate (EP, n = 8) or distilled water of equal volume (EC, n = 8). These groups completed acute high-intensity interval exercise (HIIE) after 7 days of supplementation. The acid metabolism variables were measured immediately after exercise including blood pH (pHe), base excess (BE), HCO3 -, blood lactic acid and skeletal muscle pH (pHi). The redox state was determined by measuring the oxidized coenzyme I/reduced coenzyme I (nicotinamide adenine dinucleotide [NAD+]/reduced NAD+ [NADH]) ratio and lactate/pyruvate (L/P) ratio. In addition, the activities of lactate dehydrogenase A (LDHA), hexokinase (HK), phosphofructokinase (PFK) and pyruvate kinase (PK) were determined by ELISA. Pyruvate supplementation significantly reversed the decrease of pHe, BE, HCO3 - and pHi values after HIIE (p < 0.001), while significantly increased the activities of LDHA (p = 0.048), HK (p = 0.006), and PFK (p = 0.047). Compared with the CC, the NAD+/NADH (p = 0.008) ratio and the activities of LDHA (p = 0.002), HK (p < 0.001), PFK (p < 0.001), and PK (p = 0.006) were significantly improved in EP group. This study provides compelling evidence that oral pyruvate attenuates HIIE-induced intracellular and extracellular acidification, possibly due to increased activity of LDHA, which promotes the absorption of H+ in the LDH reaction. The beneficial effects of improving the redox state and glycolysis rate were also shown. Our results suggest that pyruvate can be used as an oral nutritional supplement to buffer HIIE induced metabolic acidosis.

Abstract 479: Novel specific PKM2 inhibitor, compound 3h, induces apoptotic and autophagic cell death through Akt/mTOR signaling pathway in prostate cancer cells

Abstract Pyruvate kinase M2 (PKM2) is a key enzyme involved in the regulation of glycolysis. Although PKM2 is overexpressed in various tumor tissues, its functional role in cancer chemotherapy remains unclear. In this study, we investigated the anticancer activity of novel PKM2 inhibitors in the regulation of cell metabolism and its associated pathways in prostate cancer cells. To evaluate the molecular basis of specific PKM2 inhibitors, the interactions of compounds 3h and 3K with the PKM2 protein were assessed via molecular docking. We found that, compared to compound 3K, compound 3h exhibited a higher binding affinity for PKM2. Moreover, compound 3h significantly inhibited the pyruvate kinase activity and PKM2 expression. Cytotoxicity and colony formation assays revealed the potent anticancer activity of compound 3h against LNCaP cells. Compound 3h significantly increased the apoptotic and autophagic cell death in LNCaP cells. In addition, compound 3h induced AMPK activation along with the inhibition of mTOR/p70S6K pathway. Furthermore, compound 3h significantly inhibited glycolysis and mitochondrial respiration, as determined by analyzing extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) production. Our results revealed that compound 3h caused apoptotic and autophagic cell death by inhibiting cancer cell metabolism in LNCaP cells. Therefore, blocking glycolytic pathways using specific PKM2 inhibitors can be used to target cancer cell metabolism in PKM2-overexpressed prostate cancer cells. Citation Format: Chunxue Jiang, Tian Zheng, HwaYoung Cha, Hyung Sik Kim. Novel specific PKM2 inhibitor, compound 3h, induces apoptotic and autophagic cell death through Akt/mTOR signaling pathway in prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 479.

5589320 FOLLOW-UP RESULTS OF A PHASE 2 STUDY ASSESSING THE SAFETY AND EFFICACY OF MITAPIVAT TREATMENT, AN ORAL PYRUVATE KINASE ACTIVATOR, FOR UP TO 60 WEEKS IN SUBJECTS WITH SICKLE CELL DISEASE

Background: Sickle cell disease (SCD) is a devastating red blood cell (RBC) disorder. The root cause of SCD is polymerization of sickle hemoglobin (HbS) upon deoxygenation, resulting in poorly deformable sickled RBCs with shortened lifespan. 2,3-diphosphyglycerate (2,3-DPG), a glycolytic intermediate in RBCs, promotes deoxygenation by lowering hemoglobin (Hb)-oxygen affinity. Mitapivat (AG-348) is an oral allosteric activator of pyruvate kinase (PK), a key enzyme in RBC glycolysis generating adenosine triphosphate (ATP) and reducing 2,3-DPG levels. Aims: To report follow-up data of the safety and efficacy of mitapivat treatment in subjects with SCD (ESTIMATE study; www.trialregister.nl NL8517; EudraCT 2019-003438-18). Methods: The ESTIMATE study is a phase 2, investigator initiated, open-label study. Subjects ≥16 years with SCD (HbSS, HbS/β0 or HbS/β+), 1-10 vaso-occlusive crises (VOCs) in the prior year and/or prior SCD-related complications, a Hb level >6.1 g/dL and ≤11.1 g/dL, and no chronic transfusions are eligible and treated with mitapivat after obtaining informed content. In the 8-week dose finding period, initial dosing is 20 mg twice daily, escalated to max. 100 mg twice daily, and followed by an up to 52-week fixed dose extension period (FDEP). Primary endpoints are safety, based on adverse events (AEs), and efficacy of mitapivat treatment on RBC sickling (Point of Sickling (PoS), oxygen gradient ektacytometry). Key secondary endpoints are changes in Hb level, markers of hemolysis, ATP/2,3-DPG levels, and Hb-oxygen affinity (p50, Hemox Analyzer). Results: Until Jan. 2022, 9 subjects were treated with mitapivat. Baseline characteristics were: median age 22 years (range 16-59 years), 6/9 (67%) female, and 6/9 (67%) used hydroxyurea. 7/9 (78%) had HbSS, 1/9 (11%) had HbS/β0, and 1/9 (11%) had HbS/β+. One subject with a non-treatment related serious AE (SAE) of a urinary tract infection (grade 4), was lost to follow-up shortly after first dosing. No other SAEs or treatment-emergent AEs (TEAEs) grade ≥3 occurred. The remaining 8 subjects had a median treatment duration of 38 weeks (range 11-60 weeks). 6/8 (75%) took 100 mg mitapivat twice daily in the FDEP, and 2/8 (25%) took 50 mg mitapivat twice daily after a single dose reduction because of progressive transaminase increase. The most common TEAEs were (n>2 subjects): ALT or AST increase (both 5/9, 56%; all grade 1), and headache (4/9, 44%; grade 1-2). Three VOCs occurred: one related to excessive alcohol consumption with no need for hospitalization, and two in subjects with documented noncompliance the week before hospitalization. Mean annual VOC rate and SCD-related hospital admission days were, respectively, 1.5±1.3 and 5.9±7.1 days at baseline, and reduced to 0.5±0.7 and 1.6±3.1 days when weighting cases by follow-up duration (p=0.021 and p=0.134, respectively). Table 1 summarizes mean changes in parameters of primary and key secondary endpoints of scheduled visits in the FDEP compared to baseline. Hb level significantly increased, accompanied by a decrease in markers of hemolysis (absolute reticulocyte count, total bilirubin, lactate dehydrogenase). Mean PoS decreased, and p50, 2,3-DPG level and ATP/2,3-DPG ratio significantly improved. Conclusion: Treatment with mitapivat for up to 60 weeks in subjects with SCD showed no treatment related TEAEs grade ≥3. Improvements in anemia, markers of hemolysis, oxygen affinity, 2,3-DPG level and ATP/2,3-DPG ratio were seen.

Analysis of the Formation of Characteristic Aroma Compounds by Amino Acid Metabolic Pathways during Fermentation with Saccharomyces cerevisiae.

Amino acid metabolic pathways can have profound impacts on the activities of key enzymes in the biosynthesis of specific aroma compounds during yeast fermentation. Aroma compounds, pyruvic acid and glucose were monitored in relation to the key enzymes of leucine aminotransferase (LTR), phenylalanine aminotransferase (PAL), pyruvate kinase (PK) and acetyl-CoA in the amino acid metabolic pathways during the fermentation of simulated juice systems with added amino acids in order to explore the formation of characteristic aroma compounds. The addition of L-phenylalanine or L-leucine to the simulated juice systems significantly improved the activities of PK, PAL and LTR, and the content of acetyl-CoA, and significantly increased the concentrations of phenylethyl alcohol, octanoic acid, isoamyl acetate, phenylethyl acetate, ethyl hexanoate and ethyl caprylate during fermentation. Correlation analysis showed that there was a significant positive correlation between PAL, LTR, PK and acetyl-CoA and pyruvic acid formation. Path analysis revealed that the addition of amino acids affected the metabolism of pyruvate to alcohols, acids and esters to some extent.

Tannic acid exerts antifungal activity in vitro and in vivo against Alternaria alternata causing postharvest rot on apple fruit

Alternaria alternata is one of the main pathogens that causes black spot rot of apple fruit, mainly postharvest. Also, A. alternata can attack many other fruits and vegetables in most of the world's natural environments. In this study, the antifungal efficacy of tannic acid (TA) against A. alternata and its potential mechanisms were investigated. An in vitro bioassay suggested that 5.0 mg mL−1 TA significantly inhibited the mycelial growth and spore germination of A. alternata. The enzyme activities of hexokinase, phosphofructokinase and pyruvate kinase in glycolytic metabolism were decreased in A. alternata treated with TA. The accumulation of glucose content and the reduction of pyruvate content in A. alternata further confirmed that TA may block the glycolytic metabolism of this fungus, resulting in insufficient energy supply for cells. In addition, TA immersion treatment significantly reduced the development of lesion areas on apple fruits caused by A. alternata and delayed fruit softening during the storage period. The application of TA also increased the activity of chitinase, β-1,3-glucanase and phenylalanine ammonialyase and the contents of total phenolic compounds and flavonoids in apple fruit, indicating the activation of defense-related enzymes and phenylpropane metabolites. TA could potentially be used to prevent and control postharvest Alternaria rot by exerting antifungal activity on A. alternata and inducing resistance in apple fruit.

Impairments in SHMT2 expression or cellular folate availability reduce oxidative phosphorylation and pyruvate kinase activity

BackgroundSerine hydroxymethyltransferase 2 (SHMT2) catalyzes the reversible conversion of tetrahydrofolate (THF) and serine-producing THF-conjugated one-carbon units and glycine in the mitochondria. Biallelic SHMT2 variants were identified in humans and suggested to alter the protein’s active site, potentially disrupting enzymatic function. SHMT2 expression has also been shown to decrease with aging in human fibroblasts. Immortalized cell models of total SHMT2 loss or folate deficiency exhibit decreased oxidative capacity and impaired mitochondrial complex I assembly and protein levels, suggesting folate-mediated one-carbon metabolism (FOCM) and the oxidative phosphorylation system are functionally coordinated. This study examined the role of SHMT2 and folate availability in regulating mitochondrial function, energy metabolism, and cellular proliferative capacity in both heterozygous and homozygous cell models of reduced SHMT2 expression. In this study, primary mouse embryonic fibroblasts (MEF) were isolated from a C57Bl/6J dam crossed with a heterozygous Shmt2+/− male to generate Shmt2+/+ (wild-type) or Shmt2+/− (HET) MEF cells. In addition, haploid chronic myeloid leukemia cells (HAP1, wild-type) or HAP1 cells lacking SHMT2 expression (ΔSHMT2) were cultured for 4 doublings in either low-folate or folate-sufficient culture media. Cells were examined for proliferation, total folate levels, mtDNA content, protein levels of pyruvate kinase and PGC1α, pyruvate kinase enzyme activity, mitochondrial membrane potential, and mitochondrial function.ResultsHomozygous loss of SHMT2 in HAP1 cells impaired cellular folate accumulation and altered mitochondrial DNA content, formate production, membrane potential, and basal respiration. Formate rescued proliferation in HAP1, but not ΔSHMT2, cells cultured in low-folate medium. Pyruvate kinase activity and protein levels were impaired in ΔSHMT2 cells and in MEF cells exposed to low-folate medium. Mitochondrial biogenesis protein levels were elevated in Shmt2+/− MEF cells, while mitochondrial mass was increased in both homozygous and heterozygous models of SHMT2 loss.ConclusionsThe results from this study indicate disrupted mitochondrial FOCM impairs mitochondrial folate accumulation and respiration, mitochondrial formate production, glycolytic activity, and cellular proliferation. These changes persist even after a potentially compensatory increase in mitochondrial biogenesis as a result of decreased SHMT2 levels.

IgSF11-mediated phosphorylation of pyruvate kinase M2 regulates osteoclast differentiation and prevents pathological bone loss

Osteoclasts are primary bone-resorbing cells, and receptor-activated NF-kB ligand (RANKL) stimulation is the key driver of osteoclast differentiation. During late-stage differentiation, osteoclasts become multinucleated and enlarged (so-called “maturation”), suggesting their need to adapt to changing metabolic demands and a substantial increase in size. Here, we demonstrate that immunoglobulin superfamily 11 (IgSF11), which is required for osteoclast differentiation through an association with the postsynaptic scaffolding protein PSD-95, regulates osteoclast differentiation by controlling the activity of pyruvate kinase M isoform 2 (PKM2). By using a system that directly induces the activation of IgSF11 in a controlled manner, we identified PKM2 as a major IgSF11-induced tyrosine-phosphorylated protein. IgSF11 activates multiple Src family tyrosine kinases (SFKs), including c-Src, Fyn, and HcK, which phosphorylate PKM2 and thereby inhibit PKM2 activity. Consistently, IgSF11-deficient cells show higher PKM2 activity and defective osteoclast differentiation. Furthermore, inhibiting PKM2 activities with the specific inhibitor Shikonin rescues the impaired osteoclast differentiation in IgSF11-deficient cells, and activating PKM2 with the specific activator TEPP46 suppresses osteoclast differentiation in wild-type cells. Moreover, PKM2 activation further suppresses osteoclastic bone loss without affecting bone formation in vivo. Taken together, these results show that IgSF11 controls osteoclast differentiation through PKM2 activity, which is a metabolic switch necessary for optimal osteoclast maturation.

Berberine regulates glucose metabolism in largemouth bass by modulating intestinal microbiota.

This study examined the role of intestinal microbiota in berberine (BBR)-mediated glucose (GLU) metabolism regulation in largemouth bass. Four groups of largemouth bass (133.7 ± 1.43g) were fed with control diet, BBR (1g/kg feed) supplemented diet, antibiotic (ATB, 0.9g/kg feed) supplemented diet and BBR + ATB (1g/kg feed +0.9g/kg feed) supplemented diet for 50days. BBR improved growth, decreased the hepatosomatic and visceral weight indices, significantly downregulated the serum total cholesterol and GLU levels, and significantly upregulated the serum total bile acid (TBA) levels. The hepatic hexokinase, pyruvate kinase, GLU-6-phosphatase and glutamic oxalacetic transaminase activities in the largemouth bass were significantly upregulated when compared with those in the control group. The ATB group exhibited significantly decreased final bodyweight, weight gain, specific growth rates and serum TBA levels, and significantly increased hepatosomatic and viscera weight indices, hepatic phosphoenolpyruvate carboxykinase, phosphofructokinase, and pyruvate carboxylase activities, and serum GLU levels. Meanwhile, the BBR + ATB group exhibited significantly decreased final weight, weight gain and specific growth rates, and TBA levels and significantly increased hepatosomatic and viscera weight indices and GLU levels. High-throughput sequencing revealed that compared with those in the control group, the Chao one index and Bacteroidota contents were significantly upregulated and the Firmicutes contents were downregulated in the BBR group. Additionally, the Shannon and Simpson indices and Bacteroidota levels were significantly downregulated, whereas the Firmicutes levels were significantly upregulated in ATB and BBR + ATB groups. The results of in-vitro culture of intestinal microbiota revealed that BBR significantly increased the number of culturable bacteria. The characteristic bacterium in the BBR group was Enterobacter cloacae. Biochemical identification analysis revealed that E. cloacae metabolizes carbohydrates. The size and degree of vacuolation of the hepatocytes in the control, ATB, and ATB + BBR groups were higher than those in the BBR group. Additionally, BBR decreased the number of nuclei at the edges and the distribution of lipids in the liver tissue. Collectively, BBR reduced the blood GLU level and improved GLU metabolism in largemouth bass. Comparative analysis of experiments with ATB and BBR supplementation revealed that BBR regulated GLU metabolism in largemouth bass by modulating intestinal microbiota.

Serotonin and dopamine regulate the aggressiveness of swimming crabs (Portunus trituberculatus) in different ways

Bioamines act as a pivotal part in the regulation of aggressive behavior in animals as a type of neuroendocrine, but the patterns of how they regulate aggressiveness in crustaceans are still unclear due to species-specific responses. To determine the effects of serotonin (5-HT) and dopamine (DA) on the aggressiveness of swimming crabs (Portunus trituberculatus), we quantified their behavioral and physiological characteristics. The results showed that an injection of 5-HT at 0.5 mmol L−1 and 5 mmol L−1 could significantly enhance the aggressiveness of swimming crabs, as well as an injection of DA at 5 mmol L-1. The regulation of 5-HT and DA on aggressiveness is dose-dependent, and these two bioamines have different concentration thresholds that can trigger aggressiveness changes. 5-HT could up-regulate the 5-HTR1 gene expression and increase lactate content at the thoracic ganglion as the aggressiveness enhances, suggesting that 5-HT may activate related receptors and neuronal excitability to regulate aggressiveness. As a result of DA injection at 5 mmol L-1, lactate content in the chela muscle and hemolymph increased, glucose content in the hemolymph increased, and the CHH gene was significantly up-regulated. Pyruvate kinase and hexokinase enzyme activities in the hemolymph increased, which accelerated the glycolysis process. These results demonstrate that DA regulates the lactate cycle, which provides substantial short-term energy for aggressive behavior. Both 5-HT and DA can mediate aggressive behavior in the crab by activating calcium regulation in muscle tissue. We conclude that the enhancement of aggressiveness is a process of energy consumption, in which 5-HT acts on the central nervous system to induce aggressive behavior, and DA affects muscle and hepatopancreas tissue to provide a large amount of energy. This study expands upon the knowledge of regulatory mechanisms of aggressiveness in crustaceans and offers a theoretical foundation for enhancing crab culture management.

Effects of Dietary Steroid Saponins on Growth Performance, Serum and Liver Glucose, Lipid Metabolism and Immune Molecules of Hybrid Groupers (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu) Fed High-Lipid Diets.

High-lipid diets are attributed to excessive lipid deposition and metabolic disturbances in fish. The aim of this experiment was to investigate the effects of steroidal saponins on growth performance, immune molecules and metabolism of glucose and lipids in hybrid groupers (initial weight 22.71 ± 0.12 g) fed high-lipid diets. steroidal saponins (0%, 0.1% and 0.2%) were added to the basal diet (crude lipid, 14%) to produce three experimental diets, designated S0, S0.1 and S0.2, respectively. After an 8-week feeding trial, no significant differences were found between the S0 and S0.1 groups in percent weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio and protein deposition rate (p > 0.05). All those in the S0.2 group were significantly decreased (p < 0.05). Compared to the S0 group, fish in the S0.1 group had lower contents of serum triglyceride and low-density lipoprotein cholesterol and higher high-density lipoprotein cholesterol and glucose (p < 0.05). The activities of superoxide dismutase, catalase and glutathione peroxidase were significantly higher, and malondialdehyde contents were significantly lower in the S0.1 group than in the S0 group (p < 0.05). Hepatic triglyceride, total cholesterol and glycogen were significantly lower in the S0.1 group than in the S0 group (p < 0.05). Activities of lipoprotein lipase, total lipase, glucokinase and pyruvate kinase, and gene expression of lipoprotein lipase, triglyceride lipase and glucokinase, were significantly higher in the S0.1 group than in the S0 group. Interleukin-10 mRNA expression in the S0.1 group was significantly higher than that in the S0 group, while the expression of interleukin-6 and tumor necrosis factor-α genes were significantly lower than those in the S0 group. In summary, adding 0.1% steroidal saponins to a high-lipid diet not only promoted lipolysis in fish livers, but also activated glycolysis pathways, thus enhancing the utilization of the dietary energy of the groupers, as well as supporting the fish's nonspecial immune-defense mechanism.

Mitapivat, a pyruvate kinase activator, improves transfusion burden and reduces iron overload in β-thalassemic mice.

Not available.

Exercise Down-Regulates Hepatic Lipogenic Enzymes in Food-Deprived and Refed Rats

The effects of an acute bout of prolonged exercise on the activities of several hepatic lipogenic enzymes and the abundance of fatty acid synthase (FAS) mRNA were evaluated using a food deprivation-refeeding protocol in which diets contained 50% of the energy from either fructose or cornstarch. Food was withheld from male rats for 48 h and refed for 0, 4, 8, 12, 24 or 48 h. At each time point, half of each dietary group was subjected to a single bout of treadmill running until exhaustion and killed immediately. The other half of each group rested without food for the same amount of time before being killed. Exercise significantly decreased FAS activity by 57, 46, 10, 26 and 70% at 4, 8, 12, 24 and 48 h of refeeding, respectively, in the fructose-fed rats; and by 70 and 63% at 24 and 48 h of refeeding, respectively, in the cornstarch-fed rats. Activities of L-type pyruvate kinase and glucose 6-phosphate dehydrogenase were significantly decreased after exercise in the fructose-fed, but not cornstarch-fed rats. In rested rats, FAS mRNA abundance increased ∼fourfold above the unfed levels after 8 and 12 h of refeeding. Exercise attenuated the diet-induced increases in FAS mRNA abundance. At 8 h of refeeding, both cornstarch- and fructose-fed exercised rats had 71% (P < 0.05) of the FAS mRNA levels of their rested counterparts; at 12 h, these exercised rats showed only 46 and 27% (P < 0.05) of FAS mRNA levels compared with rested rats fed the same diet. We conclude that dietary induction of FAS activity and mRNA abundance can be inhibited by prolonged exercise, suggesting that exercise may influence FAS transcription and/or mRNA stability.

Hydrogen sulfide alleviates chilling injury by modulating respiration and energy metabolisms in cold-stored peach fruit

The respiration and energy metabolisms of refrigerated peach fruit and their relationship to chilling injury (CI) treated with hydrogen sulfide (H2S) were investigated. Results showed that H2S treatment reduced the activities of hexokinase (HK), phosphor-glucose isomerase (PGI), phosphofructokinase (PFK) and pyruvate kinase (PK), and the contents of glucose, pyruvate and nicotinamide adenine dinucleotide (hydrogen) (NAD(H)) in the Embden-Meyerhof-Parnas (EMP) and tricarboxylic acid (TCA)-cycle compared to control and hypotaurine (HT, H2S scavenger) treatment, resulting in a lower respiration rate. Meanwhile, H2S treatment increased the activities of NADK, glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) and NADP(H) contents in the pentose phosphate pathway (PPP), while HT showed the opposite results. Moreover, H2S treatment induced the higher activities of adenosine triphosphatases (ATPases), succinic dehydrogenase (SDH) and cytochrome C oxidase (CCO) along with enhanced levels of ATP and energy charge, whereas HT suppressed energy generation, which increased cell membrane permeability and malondialdehyde content. Therefore, results suggested that H2S could maintain the integrity of the membrane structure by inhibiting the respiration and substance consumption in EMP pathway, increasing the proportion of PPP and energy supply to alleviate CI in peaches.

Effect of Acute Salinity Stress on Metabolism, Antioxidant Status, and Histological Structure of Procambarus clarkii

Procambarus clarkii is a well-known invasive species with a strong environmental tolerance that has spread worldwide. Crayfish were exposed to five different salinities (0, 6, 12, 18, and 24 parts per thousand (ppt)) to determine how salinity affects physiological and histological responses. The metabolism-related enzymes pyruvate kinase (PK) and fatty acid synthase (FAS) activities of the hepatopancreas and gill tissues significantly decreased with an increase of salinity (6, 12, 18, 24 ppt), except the increase of FAS activity at 6 ppt. Salinity caused an immune disorder, as displayed by the decreased activities of lysozyme (LSZ) and superoxide dismutase (SOD). In contrast, catalase (CAT) activities showed increased activity. The heat shock protein 90 (HSP90) concentration significantly increased at 6 ppt and then significantly decreased at 24 ppt in the hepatopancreas and gill tissues. A HE section revealed that salinity stress influenced the tissue structures. High salinity (24 ppt) damaged the hepatopancreas and deformed the gills. In conclusion, P. clarkii is resistant to acute exposure to low salinities and suffers physiological damage when exposed to hyperosmotic salinities. Our study provides a valuable reference to analyse the adaptation mechanisms of crayfish in response to salinity.

Tuning liver pyruvate kinase activity up or down with a new class of allosteric modulators

The liver isoform of pyruvate kinase (PKL) has gained interest due to its potential capacity to regulate fatty acid synthesis involved in the progression of non-alcoholic fatty liver disease (NAFLD). Here we describe a novel series of PKL modulators that can either activate or inhibit the enzyme allosterically, from a cryptic site at the interface of two protomers in the tetrameric enzyme. Starting from urolithin D, we designed and synthesised 42 new compounds. The effect of these compounds on PKL enzymatic activity was assessed after incubation with cell lysates obtained from a liver cell line. Pronounced activation of PKL activity, up to 3.8-fold, was observed for several compounds at 10 μM, while other compounds were prominent PKL inhibitors reducing its activity to 81% at best. A structure-activity relationship identified linear-shaped sulfone-sulfonamides as activators and non-linear compounds as inhibitors. Crystal structures revealed the conformations of these modulators, which were used as a reference for designing new modulators.

Effect of heat and hypoxia stress on mitochondrion and energy metabolism in the gill of hard clam

Aquatic animals suffer from heat and hypoxia stress more frequently due to global climate change and other anthropogenic activities. Heat and hypoxia stress can significantly affect mitochondrial function and energy metabolism. Here, the response and adaptation characteristics of mitochondria and energy metabolism in the gill of the hard clam Mercenaria mercenaria under heat (35 °C), hypoxia (0.2 mg/L), and heat plus hypoxia stress (35 °C, 0.2 mg/L) after 48 h exposure were investigated. Mitochondrial membrane potentials were depolarized under environmental stress. Mitochondrial fusion, fission and mitophagy played a key role in maintain mitochondrion function. The AMPK subunits showed different expression under environmental stress. Acceleration of enzyme activities (phosphofructokinase, pyruvate kinase and lactic dehydrogenase) and accumulation of anaerobic metabolites in glycolysis and TCA cycle implied that the anaerobic metabolism might play a key role in providing energy. Accumulation of amino acids might help to increase tolerance under heat and heat combined hypoxia stress. In addition, urea cycle played a key role in amino acid metabolism to prevent ammonia/nitrogen toxicity. This study improved our understanding of the mitochondrial and energy metabolism responses of marine bivalves exposed to environmental stress.

Energy metabolism of juvenile scallops Nodipecten subnodosus under acute increased temperature and low oxygen availability

High temperature increases energy demand in ectotherms, limiting their physiological capability to cope with hypoxic events. The present study aimed to assess the metabolic tolerance of juvenile Nodipecten subnodosus scallops to acute hyperthermia combined with moderate hypoxia. A previous study showed that juveniles exhibited a high upper temperature limit (32 °C), but the responses of juveniles to combined hyperthermia and low dissolved oxygen are unknown. Scallops were exposed to control conditions (treatment C: 22 °C, ∼7.1 mg O2 L−1 or PO2 156.9 mmHg), acute hyperthermia under normoxia (treatment T: 30 °C, ∼6.0 mg O2 L−1 or PO2 150.9 mmHg) or acute hyperthermia plus hypoxia (treatment TH: 30 °C, ∼2.5 mg O2 L−1 or PO2 62.5 mmHg) for 18 h. In T, juveniles exhibited an enhanced oxygen consumption, together with a decrease in adenylate energy charge (AEC) and arginine phosphate (ArgP), and with no changes in metabolic enzyme activity in the muscle. In TH, scallops maintained similar AEC and ArgP levels in muscle as those observed in T treatment. This response occurred along with the accumulation of inosine monophosphate and hypoxanthine. Besides, reduced citrate synthase and pyruvate kinase activities, enhanced hexokinase activity, and a higher octopine dehydrogenase/lactate dehydrogenase ratio in the mantle indicated the onset of anaerobiosis in TH. These responses indicate that juvenile scallops showed tissue-specific compensatory responses regarding their energy balance under moderate hypoxia at high temperatures. Our results give an insight into the tolerance limit of this species to combined hyperthermia and hypoxia in its northern limit of distribution.

Aerobic respiration, biochemical composition, and glycolytic responses to ultraviolet radiation in jellyfish Cassiopea sp

The light-dependent zooxanthellate jellyfish Cassiopea sp. (the upside-down jellyfish) is invasive/exotic in many shallow and clear marine habitats, where the jellyfish might be exposed to high levels of ultraviolet radiation (UVR). Compared to other reef organisms, the sensitivity/resilience of the semi-transparent jellyfish to UVR exposure is overlooked. Therefore, we experimentally investigated the metabolic and physiological responses of Cassiopea sp. from the Red Sea to natural levels of underwater UVR following 16 days of exposure to three light treatments: 1) control group with only photosynthetically active radiation (PAR), 2) PAR+UV-B, and 3) PAR+UV-B+UV-A. While jellyfish body mass increased (by 40%) significantly in the control group, it did not increase in either of the UV treatments. However, both UV-exposed jellyfish had higher (98% to 120%) mitochondrial electron transport system (ETS) activity than the control group. Therefore, the results indicate elevated aerobic respiration rates in UV-exposed jellyfish (i.e., reflecting a higher energy cost of UVR exposure). Neither the lactate dehydrogenase (LDH) activity nor the available energy (Ea) exhibited different levels among UVR treatments compared to the control group. In contrast, pyruvate kinase activity was significantly lower (by 46%) in all UV-exposed jellyfish compared to the control group. Unchanged Ea and LDH activity combined with higher ETS activity indicates a high aerobic capacity of jellyfish, which might explain their ability to cope with UVR exposure-induced higher energy demands without inducing the onset of anaerobiosis. The results indicated that UV-A does not amplify or modulate jellyfish physiology and growth under UV-B exposure. In conclusion, the findings suggest that the jellyfish is more resilient (i.e., in terms of survival) to UVR than other cnidarians. This study on Cassiopea is the first to address its metabolic and physiological responses to UVR. Therefore, it could be used as a framework for further studies aiming to better understand jellyfish physiology.