6-phosphogluconate Dehydrogenase Activities Research Articles

Hydroxyethyl starch-based self-reinforced nanomedicine inhibits both glutathione and thioredoxin antioxidant pathways to boost reactive oxygen species-powered immunotherapy

The adaptive antioxidant systems of tumor cells, predominantly glutathione (GSH) and thioredoxin (TRX) networks, severely impair photodynamic therapy (PDT) potency and anti-tumor immune responses. Here, a multistage redox homeostasis nanodisruptor (Phy@HES-IR), integrated by hydroxyethyl starch (HES)-new indocyanine green (IR820) conjugates with physcion (Phy), an inhibitor of the pentose phosphate pathway (PPP), is rationally designed to achieve PDT primed cancer immunotherapy. In this nanodisruptor, Phy effectively depletes intracellular GSH of tumor cells by inhibiting 6-phosphogluconate dehydrogenase (6PGD) activity. Concurrently, it is observed for the first time that the modified IR820-NH2 molecule not only exerts PDT action but also interferes with TRX antioxidant pathway by inhibiting thioredoxin oxidase (TRXR) activity. The simultaneous weakening of two major antioxidant pathways of tumor cells is favorable to maximize the PDT efficacy induced by HES-IR conjugates. By virtue of the excellent protecting ability of the plasma expander HES, Phy@HES-IR can remain stable in the blood circulation and efficiently enrich in the tumor region. Consequently, PDT and metabolic modulation synergistically induced immunogenic cell death, which not only suppressed primary tumors but also stimulated potent anti-tumor immunity to inhibit the growth of distant tumors in 4T1 tumor-bearing mice.

Using Omics Techniques to Analyze the Effects of Gene Mutations and Culture Conditions on the Synthesis of β-Carotene in Pantoea dispersa

β-carotene possesses antioxidant properties and holds significant research value. In our study, we have successfully identified a strain of Pantoea dispersa MSC14 which has the capability to produce β-carotene. By incorporating corn steep liquor powder into culture medium and employing mutagenesis breeding techniques, we have successfully increased the production of β-carotene in the MSC14 strain by 13.97% and 29.22%, respectively. To gain further insights, we conducted genomic and transcriptomics analyses. These analyses revealed a significant mutation in the gndA (6-phosphogluconate dehydrogenase) gene of the mutant strain 14P9, resulting in a 33.74% decrease in 6-phosphogluconate dehydrogenase activity. Using transcriptomics analysis, we investigated the impact of this mutation on β-carotene production and explored the interconnectedness between carbon metabolism, fatty acid metabolism, amino acid metabolism, and β-carotene synthesis. The up-regulation of the trxC (Thioredoxin-2) gene, as observed in both transcriptomics results, prompted us to construct strains that overexpress trxC. This manipulation resulted in a notable 15.89% increase in β-carotene production, highlighting the significant impact of of the trxC gene on the β-carotene content of Pantoea dispersa. In conclusion, our study has successfully identified Pantoea dispersa MSC14 as a proficient producer of β-carotene. Furthermore, we have uncovered two genes implicated in the biosynthesis of β-carotene. These findings enhance our understanding of β-carotene synthesis and provide valuable guidance for carotenoid biosynthesis.

Biochemical and metabolomics analyses reveal the mechanisms underlying ascorbic acid and chitosan coating mediated energy homeostasis in postharvest papaya fruit

Papaya is a climacteric fruit that undergoes rapid ripening and quality deterioration during postharvest storage, resulting in significant economic losses. This study employed biochemical techniques and targeted metabolomics to investigate the impact of exogenous AsA + CTS application on the energy metabolism regulation of papaya fruit during postharvest storage. We found that AsA + CTS treatment significantly increased the levels of key metabolic compounds and enzymes, such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), and the energy charge, as well as the succinic acid content and the activities of succinic dehydrogenase (SDH), cytochrome c oxidase (CCO), H+-ATPase, and Ca2+-ATPase. Moreover, AsA + CTS coating augmented the nicotinamide adenine dinucleotide kinase (NADK) activity and increased the NADH and NADPH concentrations. Regarding sugar metabolism, it increased the activities of 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase and raised d-glucose-6-phosphate levels. These findings suggest that AsA + CTS coating application can mitigate the metabolic deterioration and sustain a primary metabolism homeostasis in papaya fruit by enhancing the tricarboxylic acid (TCA) cycle and pentose phosphate pathway (PPP), thereby preserving their quality attributes during postharvest storage.

Combined effects of dietary carbohydrate levels and ammonia stress on growth, antioxidant capacity and glucose metabolism in juvenile oriental river prawn (Macrobrachium nipponense).

Ammonia is a common environmental stress factor that constrains aquaculture industry development. This study evaluated the effect of carbohydrate levels and ammonia stress in oriental river prawn (Macrobrachium nipponense). The experiment had six treatments containing two water ammonia levels (0 and 5 mg/L) and three dietary carbohydrate levels (low carbohydrate diet (LCD, 10%), medium carbohydrate diet [MCD, 20%], and high carbohydrate diet [HCD, 30%]), and lasted six weeks. The results showed that the prawns fed on MCD had higher weight gain than those fed on LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower acid phosphatase and alkaline phosphatase activities during ammonia stress. Feeding the prawns on the MCD increased B cells in the hepatopancreas during ammonia stress. Interestingly, the prawns fed on MCD had significantly lower superoxide dismutase activity compared to LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower pyruvate kinase activity and pyruvate and lactic acid contents, while those fed on LCD had significantly higher succinic dehydrogenase, 6-phosphogluconic dehydrogenase, and phosphoenol pyruvate carboxykinase activities during ammonia stress. The prawns fed on the MCD increased significantly glutaminase activity and decreased the ammonia content in the serum during ammonia exposure. In addition, feeding the prawns on MCD decreased significantly the expression of apoptosis and inflammation-related genes. Taken together, the MCD supplied energy required to counteract ammonia stress, which increased growth, improved antioxidant capacity, facilitated ammonia excretion, and alleviated inflammation and apoptosis of the oriental river prawn.

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.

Magnesium Reduces Hepatic Lipid Accumulation in Yellow Catfish (Pelteobagrus fulvidraco) and Modulates Lipogenesis and Lipolysis via PPARA, JAK-STAT, and AMPK Pathways in Hepatocytes

BackgroundMagnesium influences hepatic lipid deposition in vertebrates, but the underlying mechanism is unknown. ObjectiveWe used yellow catfish and their isolated hepatocytes to test the hypothesis that magnesium influences lipid deposition by modulating lipogenesis and lipolysis. MethodsJuvenile yellow catfish (mean ± SEM weight: 3.43 ± 0.02 g, 3 mo old, mixed sex) were fed a 0.14- (low), 0.87- (intermediate) or 2.11- (high) g Mg/kg diet for 56 d. Primary hepatocytes were incubated for 48 h in control or MgSO4-containing medium with or without 2-h pretreatment with an inhibitor (AG490, GW6471, or Compound C). Growth performance, cell viability, triglyceride (TG) concentrations, and expression of enzymes and genes involved in lipid metabolism were measured. ResultsCompared with fish fed low magnesium, those fed intermediate or high magnesium had lower hepatic lipids (18%, 22%) and 6-phosphogluconate dehydrogenase (6PGD; 3.7%, 3.8%) and malic enzyme (ME; 35%, 48%) activities and greater mRNA levels of the lipolytic genes adipose triacylglyceride lipase (atgl; 82% and 1.7-fold) and peroxisome proliferator-activated receptor (ppara; 18% and 1.0-fold), respectively (P < 0.05). Relative mRNA levels of AMP-activated protein kinase (ampk) a1, ampka2, ampkb1, ampkb2, ampkg1a, ampkg1b, Janus kinase (jak) 2a, jak2b, and signal transducers and activators of transcription (stat) 3 in fish fed high magnesium were higher (24% to 3.1-fold, P < 0.05) than in those fed low or intermediate magnesium. Compared with cells incubated with MgSO4 alone, those incubated with MgSO4 and pretreated with AG490, GW6471, or Compound C had greater TG concentrations (42%, 31%, or 56%), g6pd (98%, 59%, or 51%), 6pgd (68%, 73%, or 32%) mRNA expression, and activities of G6PD (35%, 45%, or 16%) and ME (1.5-fold, 1.3-fold, or 13%), and reduced upregulation (61%, 25%, or 45%) of the lipolytic gene, atgl (P < 0.05). ConclusionsMagnesium reduced hepatic lipid accumulation in yellow catfish and the variation might be attributed to inhibited lipogenesis and increased lipolysis. PPARA, JAK-STAT, and AMPK pathways mediated the magnesium-induced changes in lipid deposition and metabolism. These results offer new insight into magnesium nutrition in vertebrates.

Adverse effects of chronic ammonia stress on juvenile oriental river prawn (Macrobrachium nipponense) and alteration of glucose and ammonia metabolism.

Ammonia is one of the common stress factors in aquaculture. However, the effect of chronic ammonia exposure in juvenile oriental river prawn (Macrobrachium nipponense) is currently unexplored. This study explored the effects of chronic ammonia on juvenile healthy oriental river prawns. Fifty prawns (0.123 ± 0.003 g) were exposed to 0, 5, and 15 mg/L total ammonia nitrogen (TAN) in triplicates for 28 days. The effects of chronic ammonia challenge were evaluated on growth, antioxidant capacity, hepatopancreas and gill morphology, and glucose and ammonia metabolism. The results showed that, the chronic ammonia exposure reduced significantly survival rate and weight gain of prawns. The prawns exposed to 15 mg/L ammonia had induced oxidative stress. However, the prawn exposed to 15 mg/L ammonia had significantly lower aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and acid phosphatase activities in the serum. Furthermore, exposure of prawns to 15 mg/L ammonia increased the activities of hexokinase, pyruvate kinase, pyruvate and lactic acid content, and glutamine synthase activity. However, the prawns exposed to 15 mg/L ammonia, reduced succinic dehydrogenase, 6-phosphogluconic dehydrogenase, phosphoenolpyruvate carboxykinase, glutamate synthase, and glutamate dehydrogenase activities but increased ammonia content in serum. The exposure of ammonia deformed lumen, damaged basement membrane and decreased secretory cells in the hepatopancreas, disordered gill epithelial and pillar cells, and caused gill filament base vacuolation. Our study indicates that chronic ammonia stress impairs growth performance, tissue morphology, induces oxidative stress, and alters glucose and ammonia metabolism in juvenile oriental river prawns.

60Coγ-ray irradiation inhibits germination of fresh walnuts by modulating respiratory metabolism and reducing energy status during storage

Postharvest germination negatively affects the development of fresh walnut industry. Our previous research showed that 60Coγ-ray irradiation with 0.3 kGy effectively inhibited the germination of fresh walnuts during storage, but the underlying mechanism in respiratory metabolism remains still unclear. The untreated and 0.3 kGy-irradiated fresh walnuts were stored at 25 °C for 24 d, and then the indicators related to respiratory metabolism in kernels and embryos were determined. The results found that the 0.3 kGy irradiation decreased the respiration intensity after 15d and completely inhibited the germination of fresh walnuts with the declined adenosine triphosphate (ATP) content and energy charge (EC) levels in kernels except from 12 to 15 d and polyphenol oxidase (PPO), cytochrome C oxidase (CCO) and ascorbic acid oxidase (AAO) activities in both tissues except the stimulated PPO activity in embryos during storage. Moreover, the irradiation promoted the glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) activities but reduced the isocitrate lyase (ICL) activity in both tissues, and decreased the nicotinamide adenine dinucleotide reduced form (NADH) content but increased the nicotinamide adenine dinucleotide phosphate reduced form (NADPH) content in kernels of fresh walnuts during storage. The irradiation induced the JRG6PDH and JR6PGDH expression levels in both tissues, as well as the JRICL and JRMS expression levels in kernels before 12 d but suppressed those after 12 d of storage. In conclusion, the germination inhibition of fresh walnuts by gamma irradiation is attributed to suppressed terminal oxidases activities, decreased ratio of Embden–Meyerhof–Parnas pathway-tricarboxylic acid cycle (EMP-TCA) and glyoxylate cycle (GAC), increased level of pentose phosphate pathway (PPP) and reduced energy status.

Effect of dietary carbohydrates on growth performance, feed efficiency and glucose metabolism in common snook (Centropomus undecimalis) and yellowtail snapper (Ocyurus chrysurus) juveniles

In recent years, interest in commercial farming of the common snook Centropomus undecimalis and yellowtail snapper Ocyurus chrysurus has been increasing in the Atlantic West; however, these species are carnivorous with a high protein requirement in the diet. A 75-day feeding trial was conducted to evaluate the effect of dietary carbohydrates on growth performance, feed efficiency, glycemic response, digestive and key liver enzymes of intermediary metabolism of C. undecimalis and O. chrysurus. Two diets were formulated to contain 51.6% protein, 7.9% lipid and 20% corn starch (High digestive carbohydrates diet, HC) or 20% wheat bran (Control diet, C). Growth performance was not affected by dietary carbohydrates in both species. The feed efficiency (FE) and protein efficiency ratio (PER) improved in C. undecimalis fed the HC diet; however, for O. chrysurus, FE and PER were not affected by the HC diet. The hepatic glycogen and whole-body lipid content increased in C. undecimalis fed the HC diet, but this did not occur in O. chrysurus. Intestinal amylase activity increased in both species with the HC diet. Blood glucose peaked in C. undecimalis six hours after being fed with the HC diet (7.8 mmol L−1), whereas the glucose peak in O. chrysurus occurred until nine hours (14.1 mmol L−1). Hepatic glucokinase (GK) activity increased with the HC diet in both fish species. However, at the molecular level, GK mRNA expression was higher in C. undecimalis fed the HC diet, but for O. chrysurus, GK mRNA expression did not differ between diets. Glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) activities increased only in C. undecimalis fed the HC diet. Pyruvate kinase (PK) increased and fructose-1, 6-bisphosphatase (FBPase) decreased in both species fed the HC diet. Alanine aminotransferase (ALT) activity was unaffected by experimental diets, but O. chrysurus showed higher activity than C. undecimalis. These results suggest that both fish species are able to adapt to a high-carbohydrate diet by reorganization carbohydrate metabolism. However, C. undecimalis showed a greater capacity to utilize and store carbohydrates than O. chrysurus.

Physiological and metabolic protein‐sparing effects of dietary lipids on common snookCentropomus undecimalis(Bloch, 1792) juveniles

Carnivorous common snook (Centropomus undecimalis) has potential for aquaculture, but its protein requirement in diet is 500 g kg−1 and the energetic role of lipids in this species has not been explored. Thus, this study investigated the ability of juvenile common snook (8.7 ± 1.2 g) to use dietary lipids as an energy source. The fish were fed four practical diets with two protein levels (400 and 500 g kg−1) associated with two lipid levels (60 and 120 g kg−1) for 12 weeks. Growth performance significantly improved with increasing dietary lipid levels, but no significant difference was observed at the protein level. Fish fed the 400:60 (protein: lipid level) diet exhibited significantly higher feed conversion ratios than those fed 400:120, 500:60 and 500:120 diets, but fish fed the 400:120 diet had significantly higher protein efficiency ratios. Juvenile common snook showed the ability to modulate digestive protease and lipase secretion when dietary protein or lipid levels were modified. The activity of alanine aminotransferase decreased as the dietary lipid level increased (p < .05), while fructose-1 and 6-bisphosphatase activities were not significantly affected by dietary protein or lipid levels. Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and pyruvate kinase activities increased as dietary protein and lipid levels increased (p < .05). The results of this study indicate that an increase in dietary lipid level has a protein-sparing effect and that the diet containing 400 g kg−1 protein with 120 g kg−1 lipid is suitable for growth and effective protein utilization in common snook.

Inhibition of downy blight and enhancement of resistance in litchi fruit by postharvest application of melatonin

Litchis are tasty fruit with economic importance. However, the extreme susceptibility of harvested litchis to litchi downy blight caused by Peronophythora litchii leads to compromised quality. This study aimed to study the effects of melatonin on postharvest resistance to P. litchii in ‘Feizixiao’ litchis. Results showed that melatonin restricted lesion expansion in litchis after P. litchi inoculation. Melatonin enhanced the activities of phenylalanine ammonia-lyase, cinnamate-4-hydroxylase and 4-hydroxycinnamate CoA ligase while promoting the accumulations of phenolics and flavonoids. Nicotinamide adenine dinucleotide phosphate content and glucose-6-phosphate dehydrogenase and 6-phosphogluconic acid dehydrogenase activities were higher in treated fruit than control fruit. Higher energy status along with elevated H+-ATPase, Ca2+-ATPase, succinate dehydrogenase and cytochrome C oxidase activities were observed in treated fruit. Ultrastructural observation showed reduced damage in mitochondria in treated fruit. The results suggest that melatonin induced resistance in litchis by modulating the phenylpropanoid and pentose phosphate pathways as well as energy metabolism..

Alterations in Enzyme Activity of Carbonic Anhydrase, 6-phosphogluconate Dehydrogenase and Thioredoxin Reductase in Rats Exposed to Doxorubicin and Morin

Objective: Carbonic anhydrase (CA), 6-phosphogluconate dehydrogenase (6PGD) and thioredoxin reductase (TrxR) enzymes are the essential biological molecules needed for metabolic processes in all living cells. This study was designed to investigate the activities of CA, 6PGD and TrxR enzymes in the brain, kidney, liver, heart and testis tissues of the rats exposed to Dox and morin. Methods: Male Wistar albino rats were randomly divided into three groups as control, morin and DOX, each of them containing 7 rats. At the end of the experimental procedure, CA, 6PGD and TrxR enzyme activities in tissues of rats were determined spectrophotometrically. Results: In our study, we observed that DOX activated CA enzyme in liver and kidney tissues while inhibiting CA enzyme in the other tissues, activated 6PGD enzyme in the kidney, liver and heart tissues, and inhibited the TrxR enzyme in all the tissues. In addition, morin activated CA enzyme in the liver tissue while inhibiting CA enzyme in the brain, heart and testis tissues. Morin activated 6PGD enzyme activity while it inhibited TrxR enzyme in all the tissues. Conclusion: The findings showed that doxorubicin and morin had similar properties in the tissues as to their effect on enzyme activities.

Exogenous phytosulfokine α (PSKα) applying delays senescence and relief decay in strawberry fruits during cold storage by sufficient intracellular ATP and NADPH availability

Herein, we employed exogenous phytosulfokine α (PSKα) for delaying senescence and lessening decay in strawberry fruits during storage at 4 °C for 18 days. Our results showed that the strawberry fruits treated with 150 nM PSKα exhibited lower expression of poly-ADP-ribose polymerase 1 (PARP1) gene, leading to a higher intracellular NAD+ availability, beneficial for a sufficient provision of intracellular NADP+ with the activity of NAD kinase (NADK). Moreover, higher activities of glucose 6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), and methylenetetrahydrofolate dehydrogenase (MTHFD) may be the reason for the sufficient intracellular availability of NADPH in strawberry fruits treated with 150 nM PSKα. In addition, strawberry fruits treated with 150 nM PSKα exhibited a sufficient availability of ATP resulted from higher activities of succinate dehydrogenase (SDH) and cytochrome c oxidase (CCO). Therefore, our results indicate that exogenous PSKα could be beneficial for delaying senescence and reducing decay in strawberry fruits during cold storage.

Effect of different carbohydrate-to-lipid ratios elicits growth, feed utilization, lipid deposition and lipogenic enzyme activity in striped catfish (Pangasianodon hypophthalmus, Sauvage, 1878) fingerlings

This study evaluated the effects of diets containing various carbohydrate-to-lipid (CHO L-1) ratios on growth performance, nutrient utilization body indices and hepatic lipogenic enzyme (malic enzyme, 6-phosphogluconate dehydrogenase and fatty acid synthase) activities. Triplicate groups of Pangasianodon hypophthalmus fingerlings were fed eight isoenergetic and isonitrogenous diets with different carbohydrate-to-lipid ratios (0.51, 0.79, 1.12, 1.79, 2.41, 3.24, 4.43 and 7.62). Higher body fat deposition and lower growth performance were observed in P. hypophthalmus fingerlings fed with high-lipid diet than those fed with high-carbohydrate diet. The fish fed the diet with 7.62 CHO L-1 ratio exhibited significantly (p<0.05) higher hepatosomatic index compared to those fed higher lipid diets (0.51 and 0.79). High dietary carbohydrate level significantly increased (p<0.05) the activities of malic, 6-phosphogluconate dehydrogenase and fatty acid synthase enzyme. Based on the second-order polynomial regression analysis of weight gain, the optimal dietary carbohydrate and lipid contents for P. hypophthalmus fingerling were 304 and 103 g kg−1, respectively, which correspond to a dietary CHO L-1 ratio of 2.95.

Dietary Glucose Increases Glucose Absorption and Lipid Deposition via SGLT1/2 Signaling and Acetylated ChREBP in the Intestine and Isolated Intestinal Epithelial Cells of Yellow Catfish

Dietary carbohydrate affects intestinal glucose absorption and lipid deposition, but the underlying mechanisms are unknown. We used yellow catfish and their isolated intestinal epithelial cells (IECs) to test the hypothesis that sodium/glucose cotransporters (SGLTs) 1/2 and acetylated carbohydrate response element binding protein (ChREBP) mediated glucose-induced changes in glucose absorption and lipid metabolism. Yellow catfish (mean±SEM weight: 4.68±0.02g, 3mo old, mixed sex) were fed diets containing 250g carbohydrates/kg from glucose (G, control), corn starch (CS), sucrose (S), potato starch (PS), or dextrin (D) for 10 wk. IECs were isolated from different yellow catfish and incubated for 24h in a control or glucose (15mM) solution with or without a 2-h pretreatment with an inhibitor [sotagliflozin (LX-4211) or tubastatin A (TBSA)]. Human embryonic kidney cells (HEK293T cells) were transfected with a Flag-ChREBP plasmid to explore ChREBP acetylation. Triglyceride (TG) and glucose concentrations and enzymatic activities were measured in the intestine and IECs of yellow catfish. They also were subjected to immunofluorescence, immunoprecipitation, qPCR, and immunoblotting. Immunoblotting and immunoprecipitation were performed with HEK293T cells. The G group had greater intestine TGs (0.99- to 2.30-fold); activities of glucose 6-phospate dehydrogenase, 6-phosphogluconate dehydrogenase, and isocitrate dehydrogenase (0.12- to 2.10-fold); and expression of lipogenic genes (0.32- to 2.34-fold) thanthe CS,PS,andDgroups. The G group had greater intestine sglt1/2 mRNA and protein expression than the CS, Sand Dgroups (0.35- to 1.12-fold and 0.40- to 4.67-fold, respectively), but lower mRNA amounts of lipolytic genes(48.6%-65.8%) than the CS and PS groups. LX-4211 alleviated the glucose-induced increase in sglt1/2 mRNA (38.2%-47.4%) and SGLT1 protein (48.0%) expression, TGs (29.3%), and lipogenic enzyme activities (27.7%-42.1%) and gene expression (38.0%-55.5%) in the IECs. TBSA promoted the glucose-induced increase in TGs (11.3%), fatty acid synthase activity (32.6%), and lipogenic gene expression (21.6%-34.4%) in the IECs and acetylated ChREBP (10.5%) in HEK293T cells. SGLT1/2 signaling and acetylated ChREBP mediated glucose-induced changes in glucose absorption and lipid metabolism in the intestine and IECs of yellow catfish.

A High-Throughput Method for Directed Evolution of NAD(P)+-Dependent Dehydrogenases for the Reduction of Biomimetic Nicotinamide Analogues.

Engineering flavin-free NAD(P)+-dependent dehydrogenases to reduce biomimetic nicotinamide analogues (mNAD+s) is of importance for eliminating the need for costly NAD(P)+ in coenzyme regeneration systems. Current redox dye-based screening methods for engineering the mNAD+ specificity of dehydrogenases are frequently encumbered by a background signal from endogenous NAD(P) and intracellular reducing compounds, making the detection of low mNAD+-based activities a limiting factor for directed evolution. Here, we develop a high-throughput screening method, NAD(P)-eliminated solid-phase assay (NESPA), which can reliably identify mNAD+-active mutants of dehydrogenases with a minimal background signal. This method involves (1) heat lysis of colonies to permeabilize the cell membrane, (2) colony transfer onto filter paper, (3) washing to remove endogenous NAD(P) and reducing compounds, (4) enzyme-coupled assay for mNADH-dependent color production, and (5) digital imaging of colonies to identify mNAD+-active mutants. This method was used to improve the activity of 6-phosphogluconate dehydrogenase on nicotinamide mononucleotide (NMN+). The best mutant obtained after six rounds of directed evolution exhibits a 50-fold enhancement in catalytic efficiency (k cat/K M) and a specific activity of 17.7 U/mg on NMN+, which is comparable to the wild-type enzyme on its natural coenzyme, NADP+. The engineered dehydrogenase was then used to construct an NMNH regeneration system to drive an ene-reductase catalysis. A comparable level of turnover frequency and product yield was observed using the engineered system relative to NADPH regeneration by using the wild-type dehydrogenase. NESPA provides a simple and accurate readout of mNAD+-based activities and the screening at high-throughput levels (approximately tens of thousands per round), thus opening up an avenue for the evolution of dehydrogenases with specific activities on mNAD+s similar to the levels of natural enzyme/coenzyme pairs.

Inhibition effects of some antidepressant drugs on pentose phosphate pathway enzymes

The glucose metabolism in the pentose cycle is essential to the source of NADPH. Deficiency of these enzymes have been linked to depression and psychotic disorders. Depression is an increasingly prevalent mental disorder which may cause loss of labor. Antidepressant drugs are commonly employed in treatments of mood disorders and anxiety treatment. The purpose of this study is to investigate the effects of aripiprazole, mirtazapine, risperidone, escitalopram and haloperidol on the activity of 6-phosphogluconate dehydrogenase (6PGD) and glucose-6-phosphate dehydrogenase (G6PD) enzymes purified from human erythrocytes. It was found that aripiprazole, mirtazapine, risperidone, escitalopram and haloperidol show effective inhibitor properties on purified G6PD and 6PGD enzymes. The IC50 values of these drugs were found in the range of 26.34 μM-5.78 mM for 6PGD and 16.26 μM-3.85 mM for G6PD. The Ki values of the drugs were found in the range of 30.21 ± 4.31 μM-4.51 ± 1.83 mM for 6PGD and 14.12 ± 3.48 μM-4.98 ± 1.14 mM for G6PD. Usage of drugs with significant biological effects may be a hazard in some conditions.

In vitro and in vivo effects of iron on the expression and activity of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase in rat spleen.

Iron is an indispensable element for vital activities in almost all living organisms. It is also a cofactor for many proteins, enzymes, and other essential complex biochemical processes. Therefore, iron trafficking is firmly regulated by Hepcidin (Hamp), which is regarded as the marker for iron accumulation. The disruption of iron homeostasis leads to oxidative stress that causes various human diseases, but this mechanism is still unclear. The aim of this study is to provide a better in vivo and in vitro understanding of how long-term iron overload affects the gene expression and activities of some antioxidant enzymes, such as glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), and glutathione reductase (GR) in the spleen. The findings of this study show that iron overload reduces the gene expression of G6pd, 6pgd, and Gr, but its actual effect was on the protein level.

Modulations of Some Carbohydrate Metabolic Enzymes by Aqueous and Ethanol Buchholzia coriacea Seed Extract in Alloxan Induced Diabetic Rats

Characterised by abnormal increase in blood glucose level, Diabetes is a metabolic disorder that is associated with complications in carbohydrate, protein and fat metabolism. In recent times, medicinal herbs have been implicated in traditional medical practice for the treatment of this ailment. Studies have shown that Buchholzia coriacea seed possesses some anti-hyperglycemic properties that may be useful in the management of diabetes. To this point, present study investigated the effect(s) of oral administration of aqueous and ethanol extracts of Buchholzia coriacea on some carbohydrate metabolism parameters in normal and alloxan-induced diabetic rats. Forty (40) adult rats of both sexes were randomly assigned into two groups (normoglycemic and hyperglycemic). While group 1 (normoglycemic) had normal control, metformin, aqueous extract (250mg/kg) and ethanol extract (250 mg/kg) treated sub-groups respectively, Group 2 (hyperglycemic) contained the diabetic control, metformin, aqueous extract (250 mg/kg), and ethanol extracted (250mg/kg) treated sub-groups dosed daily by oral gavage for 14 days. At the end of the treatment, rats were euthanized via cervical dislocation; blood samples were collected by cardiac puncture for statistical analysis. One-way analysis of variance (ANOVA) revealed that dosing with extracts had insignificant effect(s) on body weight of rats. Fasting Blood Glucose (FBG) levels were elevated before and after extracts administration. Metformin, aqueous and ethanol extracts significantly reduced (p&lt;0.05) FBG levels. Also, compared with control, total carbohydrate, liver glucose, glycosylated haemoglobin, Lactate Dehydrogenase, Isocitrate dehydrogenase, MDH, SDH, 6-phosphogluconate dehydrogenase, G6PD and CcO activities were significantly reduced (p&lt;0.05) in diabetic treated rats. Buccholzia Coriacea was therefore seen to pose hypoglycemic and glycolytic effects, regulating activities of carbohydrate metabolic enzymes. Apparently, there is a scientific merit in the use of the extract in the management of diabetes.

Impact of the Di(2-Ethylhexyl) Phthalate Administration on Trace Element and Mineral Levels in Relation of Kidney and Liver Damage in Rats.

Di(2-ethylhexyl) phthalate (DEHP) is a widely used synthetic polymer in the industry. DEHP may induce reproductive and developmental toxicity, obesity, carcinogenesis and cause abnormal endocrine function in both human and wildlife. The aim of this study was to investigate trace element and mineral levels in relation of kidney and liver damage in DEHP-administered rats. Therefore, prepubertal male rats were dosed with 0, 100, 200, and 400mg/kg/day of DEHP. At the end of the experiment, trace element and mineral levels, glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6-PGD), glutathione reductase (GR) and glutathione S-transferase (GST) enzyme activities were evaluated in the serum, liver, and kidney samples of rats. Furthermore, serum clinical biochemistry parameters, organ/body weight ratios and histological changes were investigated to evaluate impact of DEHP more detailed. Our data indicated that sodium (Na), calcium (Ca), potassium (K), lithium (Li), rubidium (Rb) and cesium (Cs) levels significantly decreased, however iron (Fe) and selenium (Se) concentrations significantly increased in DEHP-administered groups compared to the control in the serum samples. On the other hand, upon DEHP administration, selenium concentration, G6PD and GR activities were significantly elevated, however 6-PGD activity significantly decreased compared to the control group in the kidney samples. Decreased G6PD activity was the only significant change between anti-oxidant enzyme activities in the liver samples. Upon DEHP administration, aberrant serum biochemical parameters have arisen and abnormal histological changes were observed in the kidney and liver tissue. In conclusion, DEHP may induce liver and kidney damage, also result abnormalities in the trace element and mineral levels.