Phosphofructokinase Activity Research Articles

No effect of triheptanoin in patients with phosphofructokinase deficiency

Phosphofructokinase deficiency (PFKD) is a rare disorder of glycogen metabolism. The lack of phosphofructokinase activity blocks the oxidative pathway from glucose and glycogen to pyruvate. Patients suffer from myopathy, exercise intolerance, and myoglobinuria. Currently, there is no specific treatment for PFKD. We hypothesized that 2 weeks treatment with triheptanoin could improve oxidative metabolism during exercise by bypassing the blocked pyruvate generation in PFKD. The study was a randomized, double-blind, placebo-controlled crossover study. Three genetically verified patients completed two treatment periods of 14 days each with triheptanoin (0.3–1 g × kg−1 × day−1) or placebo liquid. Primary outcomes were heart rate, fatty acid and total oxidation measured via stable isotope and indirect calorimetry methodology during submaximal exercise. Triheptanoin did not improve the primary outcome heart rate during submaximal exercise compared to placebo. Palmitate oxidation was increased during submaximal exercise in one patient but did not increase in the two other patients during triheptanoin treatment. Palmitate production and palmitate utilization increased during exercise and increased to a greater extent with triheptanoin treatment in all three patients. This study suggests that triheptanoin treatment has no effect on heart rate or exercise performance despite increased palmitate production and utilization in patients with PFKD.

Sex-specific differences in the metabolic enzyme activity and transporter levels in mouse skeletal muscle during postnatal development.

Although sex-associated differences in energy metabolism in adults are well-characterized, developmental sex-specific changes in skeletal muscle metabolism are largely unknown. This study investigated sex differences in high-energy phosphate, glycolytic, and mitochondrial enzyme activities and metabolite transporter protein levels in mouse skeletal muscles during the early postnatal period (day 10), post-weaning (day 28), sexual maturity (day 56), and adult life (day 140). No significant sex-specific differences were observed on days 10 and 28, except for glucose transporter (GLUT) 4 level. The hexokinase, phosphofructokinase, and lactate dehydrogenase activities of skeletal muscle were higher and the citrate synthase, cytochrome c oxidase, and β-hydroxyacyl-CoA dehydrogenase activities were lower in female mice than those in male mice on days 56 and 140. The GLUT4 and FAT/CD36 protein levels were higher and the monocarboxylate transporter 4 level was lower in the skeletal muscles of female mice than those of male mice, particularly on days 56 and 140. At 140 days of age, the respiratory exchange ratio during treadmill running (15m/min, 60min) was lower in females than that in males, despite no sex differences at rest. In summary, sex differences were not evident in the early postnatal and post-weaning periods but became apparent after the mice reached sexual maturity. These findings indicate that sexually mature animals are a better model for investigating sex differences, particularly in the context of studying energy metabolism in mice.

Energy metabolism response of Litopenaeus vannamei to combined stress of acute cold exposure and waterless duration: Implications for physiological regulation and waterless live transport

Maintaining the homeostasis of energy metabolism is crucial for organism's stress tolerance and survival. Acute cold exposure (AC) and waterless duration (WD) represent the two predominate abiotic stressors during waterless live transport of Litopenaeus vannamei. Although previous reports have explored the physiological response of L. vannamei to combined stress AC+WD, the roles of energy metabolism response in regulation of stress tolerance remains unknown. The present study comparatively examined the variations of energy metabolism-related indicators in hemolymph (cortisol, hemocyanin, glucose and lactate), hepatopancreas and muscle tissues (levels of lactate and glycogen, activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and adenosine triphosphatase (ATPase), and ATP levels). Combined stress significantly disturbed the homeostasis of energy metabolism with the increase in levels of hemocyanin, glucose and lactate, and decrease in glycogen and ATP content (P<0.05). In addition, the activities of HK, PFK, PK, and SDH initially elevated and then decreased with the prolongation of combined stress from 3h to 9h duration, while the activity of lactate dehydrogenase (LDH) remained gradual elevation and ATPase activity decreased in a duration time dependent manner throughout the experiment. These alterations revealed that exposure to combined stress could accelerate anaerobic metabolism at initial stage and inhibit aerobic metabolism in a duration time-dependent manner, following with the reduction of energy biosynthesis and the disturbance of energy metabolism equilibrium. On the other hand, the progressive impairment on hepatopancreas tissue was observed under combined stress. In summary, the deficiency of ATP supply and histopathological injures on hepatopancreas tissue might the underlying mechanisms inducing mortality of L. vannamei during live transport.

Physiological Biochemistry-Combined Transcriptomic Analysis Reveals Mechanism of Bacillus cereus G2 Improved Salt-Stress Tolerance of Glycyrrhiza uralensis Fisch. Seedlings by Balancing Carbohydrate Metabolism.

Salt stress severely threatens the growth and productivity of Glycyrrhiza uralensis. Previous results found that Bacillus cereus G2 enhanced several carbohydrate contents in G. uralensis under salt stress. Here, we analyzed the changes in parameters related to growth, photosynthesis, carbohydrate transformation, and the glycolysis Embden-Meyerhof-Parnas (EMP) pathway-tricarboxylic acid (TCA) cycle by G2 in G. uralensis under salt stress. Results showed that G2 helped G. uralensis-accumulating photosynthetic pigments during photosynthesis, which could further increase starch, sucrose, and fructose contents during carbohydrate transformation. Specifically, increased soluble starch synthase (SSS) activity caused to higher starch content, which could induce α-amylase (AM) and β-amylase (BM) activities; increased sucrose content due to the increase of sucrose synthase (SS) activity through upregulating the gene-encoding SS, which decreased cell osmotic potential, and consequently, induced invertase and gene-encoding α-glucosidase that decomposed sucrose to fructose, ultimately avoided further water loss; increased fructose content-required highly hexokinase (HK) activity to phosphorylate in G. uralensis, thereby providing sufficient substrate for EMP. However, G2 decreased phosphofructokinase (PFK) and pyruvate kinase (PK) activities during EMP. For inducing the TCA cycle to produce more energy, G2 increased PDH activity that enhanced CA content, which further increased isocitrate dehydrogenase (ICDH) activity and provided intermediate products for the G. uralensis TCA cycle under salt stress. In sum, G2 could improve photosynthetic efficiency and carbohydrate transformation to enhance carbohydrate products, thereby releasing more chemical energy stored in carbohydrates through the EMP pathway-TCA cycle, finally maintain normal life activities, and promote the growth of G. uralensis under salt stress.

Glucose metabolism in amyotrophic lateral sclerosis: it is bitter-sweet.

Glucose metabolism in amyotrophic lateral sclerosis: it is bitter-sweet.

Microplastics boost the accumulation of tetrabromobisphenol A in a commercial clam and elevate corresponding food safety risks

Marine bivalve molluscs are one of the primary seafood for consumers. Inhabiting terrigenous pollutant-convergent coastal areas and feeding through seawater filtration, edible bivalves are subjected to waterborne emerging pollutants such as microplastics (MPs) and tetrabromobisphenol A (TBBPA). Nevertheless, the potential risks of consuming MP-TBBPA contaminated seafood are still largely unknown. With that, accumulation of TBBPA with and without the presence of MPs in a commercial bivalve species, blood clam (Tegillarca granosa), was determined in the present study. Meanwhile, corresponding target hazard quotients (THQs) as well as margins of exposure (MoEs) were estimated to evaluate the potential health risks for clam consumers. Furthermore, the impacts of pollutants accumulation on the detoxification process and energy supply were analysed. The data obtained demonstrated that MPs aggravate the accumulation of TBBPA in clams, leading to elevated potential food safety risks (indicated by higher THQ values and lower MoE values) for consumers. In addition, the in vivo contents of CYP1A1 and UDP-glucuronosyltransferase, the enzymatic activity of glutathione-S-transferase, and the expression levels of five detoxification-related genes were all dramatically suppressed by MP-TBBPA. Furthermore, clams exposed to MP-TBBPA had significantly lower adenosine triphosphate contents and lower pyruvate kinase and phosphofructokinase activities. These results indicated that the aggravation of TBBPA accumulation may be due to the hence disruption of detoxification process and limited energy available for detoxification.

Creatine nitrate supplementation strengthens energy status and delays glycolysis of broiler muscle via inhibition of LKB1/AMPK pathway

This study aimed to evaluate the effects of dietary creatine nitrate (CrN) on growth performance, meat quality, energy status, glycolysis, and related gene expression of liver kinase B1/AMP-activated protein kinase (LKB1/AMPK) pathway in Pectoralis major (PM) muscle of broilers. A total of 240 male Arbor Acres broilers (28-day-old) were randomly allocated to one of 5 dietary treatments: the basal diet (control group), and the basal diets supplemented with 600 mg/kg guanidinoacetic acid (GAA), 300, 600, or 900 mg/kg CrN (identified as GAA600, CrN300, CrN600, or CrN900, respectively). We found that dietary GAA and CrN supplementation for 14 d from d 28 to 42 did not affect broiler growth performance, carcass traits, and textural characteristics of breast muscle. GAA600, CrN600, and CrN900 treatments increased pH24h and decreased drip loss of PM muscle compared with the control (P < 0.05). The PM muscles of CrN600 and CrN900 groups showed higher glycogen concentration and lower lactic acid concentration accompanied by lower activities of phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) (P < 0.05). Simultaneously, GAA600 and all CrN treatments increased concentration of muscle creatine, phosphocreatine (PCr) and ATP, and decreased AMP concentration and AMP/ATP ratio (P < 0.05). Meanwhile, the concentrations of muscle creatine, PCr, and ATP were increased linearly, while muscle AMP concentration and AMP/ATP ratio were decreased linearly and quadratic as the dose of CrN increased (P < 0.05). GAA600, CrN600, and CrN900 treatments upregulated mRNA expression of CreaT in PM muscle, and CrN600 and CrN900 treatments downregulated GAMT expression in liver and PM muscle compared with the control or GAA600 groups (P < 0.05). The mRNA expression of muscle LKB1, AMPKα1, and AMPKα2 was downregulated linearly in response to the increasing CrN level (P < 0.05). Overall, CrN showed better efficacy on strengthening muscle energy status and improve meat quality than GAA at the some dose. These results indicate that CrN may be a potential replacement for GAA as a new creatine supplement.

Moderate-intensity training in hypoxia improves exercise performance and glycolytic capacity of skeletal muscle in horses.

We investigated whether moderate‐intensity training of horses in moderate hypoxia for 4 weeks elicits greater adaptations in exercise performance, aerobic capacity, and glycolytic/oxidative metabolism in skeletal muscle compared to normoxic training. In a randomized crossover study design, seven untrained Thoroughbred horses (5.9 ± 1.1 years, 508 ± 9 kg) completed 4 weeks (3 sessions/week) of two training protocols consisting of 3‐min cantering at 70% of maximal oxygen consumption (V˙O2max) in hypoxia (HYP; F IO2 = 14.7%) and normoxia (NOR; F IO2 = 21.0%) with a 4‐month washout period. Normoxic incremental exercise tests (IET) were conducted before and after training. Biopsy samples were obtained from the middle gluteal muscle before IET and monocarboxylate transporter (MCT) protein expression and glycolytic/mitochondrial enzyme activities were analyzed. Data were analyzed using mixed models (p < 0.05). Running speed was 7.9 ± 0.2 m/s in both groups and arterial oxygen saturation during training in NOR and HYP were 92.9 ± 0.9% and 75.7 ± 3.9%, respectively. Run time in HYP (+9.7%) and V˙O2max in both groups (NOR, +6.4%; HYP, +4.3%) at IET increased after 4 weeks of training. However, cardiac output, arterial‐mixed venous O2 difference, and hemoglobin concentration at exhaustion were unchanged in both conditions. While MCT1 protein and citrate synthase activity did not increase in both conditions after training, MCT4 protein (+13%), and phosphofructokinase activity (+42%) increased only in HYP. In conclusion, 4 weeks of moderate‐intensity hypoxic training improves exercise performance and glycolytic capacity of skeletal muscle in horses.

Tank bottom area influences the growth, molting, stress response, and antioxidant capacity of juvenile mud crab Scylla paramamosain

An 8-week trial was performed to determine the effects of different tank bottom areas (TBA) on growth performance, stress response, and antioxidant capacity of juvenile mud crab Scylla paramamosain. Four treatments in triplicates with different k-value (k = A/CW2, where A is the bottom area (mm2) of the tank, and CW is the carapace width (mm) of the juvenile crab), i.e., k-value = 4.42 (K4), 17.67 (K18), 39.76 (K40), and 70.68 (K70), were used in this experiment. The results showed no significant difference in survival rate (83.33–96.67%) was found, but the K40 group had the best growth performance among all the treatments (P < 0.05). With the decrease of TBA, the level of ecdysone and ecdysone receptor (EcR) gene expression decreased, while the molt inhibitory hormone (MIH) gene expression increased. As a result, prolonged molting intervals and reduced weight gain rates were observed in K4 and K18. Higher cortisol, lactic acid, and lower glycogen content were also observed in K4 (P < 0.05), accompanied by higher phosphofructokinase (PFK) and pyruvate kinase (PK) enzyme activities (P < 0.05). On the contrary, lower catalase (CAT) and higher malondialdehyde (MDA) were observed in the K4 and K18 groups. The results demonstrated that TBA significantly impacted the growth performance of S. paramamosain, and the effects could be mediated through hormonal, glycometabolism, and antioxidant pathways. Notably, the optimal TBA was estimated to be 29.23–59.19 (k-value) according to the quadratic polynomial regression analysis of WGR, and the oversized tank could not give extra benefits in terms of growth performance. Therefore, TBA should be deliberated when designing crab culture facilities.

The key enzymes and flavor precursors involved in formation of characteristic flavor compounds of low-salt fermented common carp (Cyprinus carpio L.)

Unique flavor is the key quality of fermented fish. In this study, the precursors and key enzymes related to the flavor of Suanyu were analyzed. The results showed that 17 kinds of free amino acids and 22 kinds of free fatty acids were detected during the fermentation. According to variable importance for predictive components values (VIP), amino acids including aspartate, phenylalanine, methionine, glutamic acid, leucine, valine, isoleucine, cysteine and fatty acids including C16:0, C20:5, C20:4, C18:1, C22:6, C18:0 and C16:1 were screened as key precursors affecting the characteristic flavor of Suanyu. Moreover, the changes of enzyme activities in different metabolic processes of flavor were determined. Phosphofructokinase (PFK), pyruvate kinase (PK), lipoxygenase (LOX), hydroperoxidase lyase (HPL) and alcohol acyltransferase (AAT) activities reached the maximum in the first week of fermentation. However, hexokinase (HK), branched chain amino acid transferase (BCAT), aromatic amino acid transferase (ARAT) and esterase activities significantly decreased with the fermentation. Furthermore, ARAT, HPL, esterase, BCAT, AAT and HK were screened as the key enzymes that affected the characteristic flavor of Suanyu. All these results indicated that the control of key flavor precursors and enzymes during fermentation was a potential method to improve the flavor of fermented fish.

Changes of Key Rate-Limiting Enzyme Activity in Glucose Metabolism After Cardiopulmonary Resuscitation.

To investigate the activity of key rate-limiting enzymes of glucose metabolism after restoration of spontaneous circulation (ROSC), to explore the potential pathophysiological mechanism of impaired myocardial energy metabolism after cardiopulmonary resuscitation (CPR). Twenty-one male Sprague-Dawley rats were randomized into three experimental groups assigned in accordance with different observation times after ROSC: Sham, instrumented rats without induced cardiac arrest or resuscitation; post-resuscitation (PR2 h); PR24 h. In these groups, CPR, including precordial compressions and synchronized mechanical ventilation, was initiated 6 min after asphyxia-induced cardiac arrest. Hearts were harvested after ROSC and samples were used to detect high-energy phosphate and glucose metabolic enzyme activity. Compared with sham, the contents of phosphocreatine and adenosine triphosphate reduced in the PR2 h group, while remained unchanged in the PR24 h group. Activities of hexokinase and pyruvate kinase did not change after ROSC. Phosphofructokinase activity decreased only in the PR24 h group. Activities of pyruvate dehydrogenase and citrate synthase fell in PR2 h group and recovered in the PR24 h group. However, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase activities fell in the PR2 h group, but did not recover in the PR24 h group. Lowered key rate-limiting enzymes activity in glucose metabolism resulted in impairment of energy production in the early stage of ROSC, but partially recovered in 24 h. This process has a role in the mechanism of impaired myocardial energy metabolism after CPR. This investigation might shed light on new strategies to treat post resuscitation myocardial dysfunction.

Dietary chromium modulates glucose homeostasis and induces oxidative stress in Pacific white shrimp (Litopenaeus vannamei).

While chromium (Cr) has been recognized as an essential nutrient for all animals, and dietary supplementation can be beneficial, it can also be toxic. The present study aimed to investigate the contrasting effects of dietary chromium in Pacific white shrimp Litopenaeus vannamei. Five experimental diets were formulated to contain Cr at levels of 0.82 (Cr0.82, unsupplemented diet), 1.01 (Cr1.01), 1.22 (Cu1.22), 1.43 (Cr1.43) and 1.63 (Cr1.63) mg/kg and were fed to shrimp for 8 weeks. Highest weight gain was recorded in shrimp fed the diet containing 1.22 mg/kg Cr. Shrimp fed the diet containing the highest level of Cr (1.63 mg/kg) showed the lowest weight gain and clear signs of oxidative stress and apoptosis as evidenced by higher levels of H2O2, malondialdehyde and 8-hydroxydeoxyguanosine, and expression of caspase 2, 3, 5, and lower contents of total and oxidized glutathione, and expression of Cu/Zn sod, cat, gpx, mt, bcl2. Chromium supplementation promoted glycolysis and inhibited gluconeogenesis as shown by increased activities of hexokinase, phosphofructokinase and pyruvate kinase, and reduced activity of phosphoenolpyruvate carboxykinase in shrimp fed the diet containing 1.43 mg/kg Cr. Shrimp fed the diet with 1.63 mg/kg Cr had lowest contents of crustacean hyperglycemic hormone and insulin like peptide in hemolymph. Expression of genes involved in insulin signaling pathway and glycose metabolism including insr, irs1, pik3ca, pdpk1, akt, acc1, gys, glut1, pk, hk were up-regulated, and foxO1, gsk-3β, g6pc, pepck were down-regulated in shrimp fed the diets supplemented with Cr. This study demonstrated that optimum dietary supplementation of Cr had beneficial effects on glucose homeostasis and growth, whereas excess caused oxidative damage and impaired growth. The results contribute to our understanding of the biological functions of chromium in shrimp.

Wogonin inhibits the growth of HT144 melanoma via regulating hedgehog signaling-mediated inflammation and glycolysis

Hedgehog (Hh) signaling has been proved to be closely associated with the occurrence of melanoma. Wogonin is one of the active components of flavonoids that extracts from Scutellariae radix. Previous studies showed that wogonin could inhibit the invasion and migration of B16F10 cells, and suppress the synthesis of melanin in A375 melanoma cells. However, the regulatory effects of Hh signaling in wogonin against melanoma and its potential mechanisms remain largely unknown. The present study aimed to investigate the effect of wogonin on the growth of HT144 melanoma, and to elucidate the role of Hh signaling in wogonin-induced antitumor effects by focusing on inflammation and glycolysis regulation. Wogonin inhibited the proliferation, colony formation and tumor growth of HT144 melanoma cells. Wogonin showed strong anti-inflammatory effect in HT144 melanoma, as shown by the decreased levels of pro-inflammatory factors, the increased level of anti-inflammatory factor and the decreased expression of inflammatory cytokines. Wogonin decreased the glucose consumption and the production of lactic acid and ATP, and decreased the activities of hexokinase (HK), phosphofructokinase(PFK) and pyruvate kinase (PK), and further inhibited the expression of monocarboxylate transporter 1 (MCT-1), MCT-4 and glucosecotransporter-1 (GLUT1), showing potent anti-glycolysis effect against HT144 melanoma. Wogonin inhibited the patched and Smo expression while increased Hhip expression in HT144 cells, suggesting that wogonin blocked the Hh signaling in HT144 cells. The Hh signaling inhibitor cyclopamine, like wogonin, inhibited the colony formation of HT144 cells, however, the inhibitory effect of wogonin on colony formation of HT144 cells was abrogated by the Hh signaling agonist SAG. In addition, SAG abrogated the inhibitory effect of wogonin on the secretion of inflammatory factors and the expression of inflammatory cytokines. Furthermore, SAG abrogated the inhibitory effect of wogonin on several key molecules controlling glycolysis. Overall, these findings suggested that the anti-tumor effect of wogonin can be attributed to the inhibition of Hh signaling-mediated regulation of inflammation and glycolysis in HT144 melanoma.

The \u03b4 subunit of F1Fo-ATP synthase is required for pathogenicity of Candida albicans

Fungal infections, especially candidiasis and aspergillosis, claim a high fatality rate. Fungal cell growth and function requires ATP, which is synthesized mainly through oxidative phosphorylation, with the key enzyme being F1Fo-ATP synthase. Here, we show that deletion of the Candida albicans gene encoding the δ subunit of the F1Fo-ATP synthase (ATP16) abrogates lethal infection in a mouse model of systemic candidiasis. The deletion does not substantially affect in vitro fungal growth or intracellular ATP concentrations, because the decrease in oxidative phosphorylation-derived ATP synthesis is compensated by enhanced glycolysis. However, the ATP16-deleted mutant displays decreased phosphofructokinase activity, leading to low fructose 1,6-bisphosphate levels, reduced activity of Ras1-dependent and -independent cAMP-PKA pathways, downregulation of virulence factors, and reduced pathogenicity. A structure-based virtual screening of small molecules leads to identification of a compound potentially targeting the δ subunit of fungal F1Fo-ATP synthases. The compound induces in vitro phenotypes similar to those observed in the ATP16-deleted mutant, and protects mice from succumbing to invasive candidiasis. Our findings indicate that F1Fo-ATP synthase δ subunit is required for C. albicans lethal infection and represents a potential therapeutic target.

Influence of resistance exercise on cardiac remodeling and soleus muscle of infarcted rats

Abstract Introduction Resistance exercise (RE) provides several benefits for healthy individuals. However, its effects during heart failure are unclear. The aim of this study was to evaluate the effects of RE on functional capacity, cardiac remodeling, and soleus muscle molecular and biochemical features in rats with myocardial infarction (MI). Methods Three months after MI or simulated surgery (Sham), Wistar rats were divided into three groups: Sham (n=14), MI (n=13), and MI subjected to RE (RE-MI, n=14). Exercised rats trained 3 times a week performing four climbs with progressive loads on a ladder over 12 weeks. Functional capacity was assessed by maximum carrying capacity test in the ladder and exercise tolerance test in treadmill. Echocardiogram was performed at the end of the study. Energy metabolism and antioxidant enzyme activities were assessed by spectrophotometry in the soleus muscle. Oxidative stress markers were analyzed in soleus muscle (lipid hydroperoxide) or serum (malondialdehyde and protein carbonylation). Protein expression of insulin type-1 growth factor-like pathway, protein kinase B, and rapamycin target complex was analyzed by Western-blot. Statistical analysis: ANOVA and Bonferroni or Dunn, Student's t and Goodman tests; p&amp;lt;0.05. Results Mortality was higher in MI than Sham. Infarction size did not differ between groups. Resistance exercise increased maximum load carrying capacity, without changing functional capacity or cardiac remodeling. Catalase activity was lower in MI than Sham and glutathione peroxidase activity was lower in MI than Sham and RE-MI. Protein carbonylation was higher in RE-MI than MI. Energy metabolism did not differ between groups, except for lower phosphofructokinase activity in RE-MI than MI. Expression of p70s6K, p-FoxO3a, and p-FoxO3a-to-FoxO3a ratio was lower, and p-p70s6K-to-p70s6K ratio was higher in MI than Sham. Conclusion The practice of resistance exercise is safe, attenuates mortality, and improves maximum load carrying capacity regardless of changes in cardiac remodeling in infarcted rats. In soleus muscle, resistance exercise preserves phosphofructokinase and antioxidant enzyme activity and expression of the proteins involved in muscle trophism. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): CNPq UNESP

Coping with extremes: lowered myocardial phosphofructokinase activities and glucose content but increased fatty acids content in highland Eurasian Tree Sparrows

BackgroundEfficient and selective utilization of metabolic substrates is one of the key strategies in high-altitude animals to cope with hypoxia and hypothermia. Previous findings have shown that the energy substrate utilization of highland animals varies with evolutionary history and phylogeny. The heart is a proxy for the cardiopulmonary system, and the metabolic substrate utilization in the myocardium is also under the strong selective pressure of chronically hypoxic and hypothermic environments. However, little information is available on the physiological adjustments in relation to metabolic substrate utilization in the myocardium for coping with high-altitude environments.MethodsWe compared the metabolic enzyme activities, including hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), citrate synthase (CS), carnitine palmitoyl transferase 1 (CPT-1), lactic dehydrogenase (LDH), and creatine kinase (CK), and metabolic substrate contents including glucose (Glu), triglyceride (TG), and free fatty acid (FFA) in the myocardium of a typical human commensal species, Eurasian Tree Sparrows (Passermontanus) between the Qinghai-Tibet Plateau (the QTP, 3230 m) and low altitude population (Shijiazhuang, 80 m), and between sexes.ResultsAmong the seven metabolic enzymes and three substrates investigated, we identified no significant differences in PK, CPT-1, HK, CS, LDH, and CK activities and TG content of the myocardium between high and low altitude populations. However, the QTP sparrows had significantly lower Glu content and PFK activities but higher FFA content relative to their lowland counterparts. In addition, male sparrows had higher myocardial HK and CS activities relative to females, independent of altitude.ConclusionsOur results showed that the QTP sparrows elevated fatty acid utilization rather than glucose preference in the myocardium relative to lowland counterpart, which contributes to uncovering both the physiological adjustments for adapting to the extreme conditions of the QTP, intraspecifically.

In vivo deep network tracing reveals phosphofructokinase-mediated coordination of biosynthetic pathway activity in the myocardium

Glucose metabolism comprises numerous amphibolic metabolites that provide precursors for not only the synthesis of cellular building blocks but also for ATP production. In this study, we tested how phosphofructokinase-1 (PFK1) activity controls the fate of glucose-derived carbon in murine hearts in vivo. PFK1 activity was regulated by cardiac-specific overexpression of kinase- or phosphatase-deficient 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase transgenes in mice (termed GlycoLo or GlycoHi mice, respectively). Dietary delivery of 13C6-glucose to these mice, followed by deep network metabolic tracing, revealed that low rates of PFK1 activity promote selective routing of glucose-derived carbon to the purine synthesis pathway to form 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR). Consistent with a mechanism of physical channeling, we found multimeric protein complexes that contained phosphoribosylaminoimidazole carboxylase (PAICS)—an enzyme important for AICAR biosynthesis, as well as chaperone proteins such as Hsp90 and other metabolic enzymes. We also observed that PFK1 influenced glucose-derived carbon deposition in glycogen, but did not affect hexosamine biosynthetic pathway activity. These studies demonstrate the utility of deep network tracing to identify metabolic channeling and changes in biosynthetic pathway activity in the heart in vivo and present new potential mechanisms by which metabolic branchpoint reactions modulate biosynthetic pathways.

Effect of key enzyme inhibition of glycolysis on synovial fibroblasts in Rheumatoid Arthritis

Objective: To observe effects of glycolysis on human rheumatoid arthritis Fibroblast-like synoviocytes (HFLS-RA) by inhibiting glycolysis. Methods: Hexokinase inhibitor (3-bromopyruvate, 3-BrPa), 6-phosphofructokinase 1 inhibitor citric acid and pyruvate kinase inhibitor shikonin were applied to HFLS-RA respectively. Cell count 8 Kit detects cell proliferation activity, the activity of hexokinase, 6-phosphofructokinase 1, and pyruvate kinase, as well as the cellular glucose, lactate and ATP content were detected by kits, and the ELISA kit detects the expression of cellular inflammatory factors TNF-α and TGF-β. Results: 10 μg/mL 3-BrPa, 160 μg/mL citric acid and 5 μg/mL shikonin significantly inhibited cell proliferation activity (P&lt;0.001); and significantly inhibited HFLS-RA hexokinase and fructose 6-phosphate Kinase 1 and pyruvate kinase activity; Glucose, lactate and ATP content decreased; TNF-α expression decreased, while TGF-β expression increased. Conclusion: This study explored the changes in glucose metabolism and the expression of inflammatory factors in HFLS-RA by inhibiting the key enzymes of glycolysis, further confirming the important role of glycolysis in HFLS-RA, and laying a theoretical basis for a deep understanding of the pathogenesis of RA.

An early-postmortem metabolic comparison among three extreme acute heat stress temperature settings in chicken breast muscle.

Normally, preslaughter acute heat stress could accelerate postmortem glycolysis and impair chicken breast (pectoralis major muscle) quality. However, previous studies indicated that it might be different when the acute heat stress temperature rises to an extreme range (above 35°C). Therefore, this study's objectives were to compare the pH decline, glycolytic enzyme activity, and AMP-activated protein kinase (AMPK) phosphorylation at early postmortem among three extreme acute heat stress temperature settings: a control group (36°C) and two experimental groups (38°C and 40°C). Although the temperature did not affect glycogen phosphorylase a and pyruvate kinase activity, there was a decrease in pH decline rate, phosphofructokinase-1 activity, and phospho-AMPK-α[Thr172] within 4h postmortem when temperature increased from 36 to 40°C. Temperature also affected hexokinase activity, with the 36°C-group having the highest activity. The results of the current study, for the first time, indicated that postmortem metabolic rate in chicken breast muscle could be changed by acute heat stress temperature setting at extreme range.

Modulation of growth, antioxidant status, hepatopancreas morphology, and carbohydrate metabolism mediated by alpha-lipoic acid in juvenile freshwater prawns Macrobrachium nipponense under two dietary carbohydrate levels

Carbohydrate is commonly used in aquatic organism as the cheapest energy source. However, most aquatic animals are low efficient in utilizing dietary carbohydrate for energy provision. Therefore, improving the utilization efficiency of high carbohydrate by aquatic organisms is important for sustainable aquaculture development. Alpha-lipoic acid (α-LA) is a cofactor of key enzymes involved in glucose metabolism. However, its role in regulation of carbohydrate metabolism in aquatic animals is currently unknown. This study investigated the effects of α-LA on growth, antioxidant status, hepatopancreas morphology, and carbohydrate metabolism of freshwater prawns Macrobrachium nipponense under two dietary carbohydrate levels. Six diets were formulated using casein and fish meal as the protein sources, fish oil and soybean oil as the lipid sources, and corn starch as the carbohydrate source, at two carbohydrate levels (15% and 30%) and three concentrations of α-LA supplementation (0, 700, and 1400 mg/kg). Each experimental diet was fed to juvenile prawns (mean weight 0.050 ± 0.003 g), twice daily to apparent satiation, in five replicates. In the 15% (low)-carbohydrate groups, weight gain was significantly reduced in prawns fed 1400 mg/kg α-LA compared with 0 or 700 mg/kg α-LA, but there was no significant effect in the 30% (high)-carbohydrate groups. Survival was significantly higher among prawns fed 700 mg/kg α-LA compared with 0 or 1400 mg/kg α-LA regardless of the carbohydrate level. The levels of dietary carbohydrate and α-LA also significantly affected the activities of key enzymes involved in glycolysis and the citric acid cycle. The activities of hexokinase and pyruvate kinase activities were highest in prawns fed l400 mg/kg α-LA, in both the low- and high-carbohydrate groups. Supplementation with 700 or 1400 mg/kg α-LA significantly increased the activities of 6-phosphofructokinase, pyruvate dehydrogenase, and succinate dehydrogenase regardless of the carbohydrate level. The level of malondialdehyde decreased significantly with increased α-LA supplementation; in contrast, the levels of superoxide dismutase, glutathione peroxidase, and glutathione were significantly higher in prawns fed 700 or 1400 mg/kg α-LA when compare with those fed no α-LA supplement. Dietary α-LA did not affect the overall morphology of the hepatopancreas, but 1400 mg/kg α-LA reduced the number of B (blister-like) cells. Thus, 700 mg/kg α-LA supplementation under either dietary carbohydrate level could improve the antioxidant status, carbohydrate metabolism, and morphology of the hepatopancreas in juveniles of M. nipponense. We conclude that α-LA supplementation can be used to improve the efficiency of carbohydrate utilization when a 30% carbohydrate diet is used for this prawn.