Glycogen Phosphorylase Activity Research Articles

Synthesis, Kinetic and Conformational Studies of 2-Substituted-5-(β-d-glucopyranosyl)-pyrimidin-4-ones as Potential Inhibitors of Glycogen Phosphorylase.

Dysregulation of glycogen phosphorylase, an enzyme involved in glucose homeostasis, may lead to a number of pathological states such as type 2 diabetes and cancer, making it an important molecular target for the development of new forms of pharmaceutical intervention. Based on our previous work on the design and synthesis of 4-arylamino-1-(β-d-glucopyranosyl)pyrimidin-2-ones, which inhibit the activity of glycogen phosphorylase by binding at its catalytic site, we report herein a general synthesis of 2-substituted-5-(β-d-glucopyranosyl)pyrimidin-4-ones, a related class of metabolically stable, C-glucosyl-based, analogues. The synthetic development consists of a metallated heterocycle, produced from 5-bromo-2-methylthiouracil, in addition to protected d-gluconolactone, followed by organosilane reduction. The methylthio handle allowed derivatization through hydrolysis, ammonolysis and arylamine substitution, and the new compounds were found to be potent (μM) inhibitors of rabbit muscle glycogen phosphorylase. The results were interpreted with the help of density functional theory calculations and conformational analysis and were compared with previous findings.

Changes in glycolytic and mitochondrial protein profiles regulates postmortem muscle acidification and oxygen consumption in dark-cutting beef

Dark-cutting beef is a condition in which beef fails to have a characteristic bright-red color when the cut surface is exposed to oxygen. However, the mechanistic basis for this occurrence is not clear. Protein expression profiles were compared between dark-cutting and normal-pH beef using LC-MS/MS-based proteomics. Mass spectrometry analysis identified 1162 proteins in the proteomes of dark-cutting and normal-pH beef. Of these, 92 proteins had significant changes in protein abundance between dark-cutting versus normal-pH beef. In dark-cutting beef, 25 proteins were down-regulated, including enzymes related to glycogen metabolism, glucose homeostasis, denovo synthesis of adenosine monophosphate (AMP), and glycogen phosphorylase activity. In comparison, 27 proteins were up-regulated in dark-cutting beef related to oxidation-reduction processes, muscle contraction, and oxidative phosphorylation. Down-regulation of glycogenolytic proteins suggests decreased glycogen mobilization and utilization, while the up-regulation of mitochondrial transport chain proteins indicates a greater capacity to support mitochondrial respiration in dark-cutting beef. These results showed that changes in proteins involved in glycogenolysis and mitochondrial electron transport would promote the development of high-pH and greater oxygen consumption, respectively; thus limiting myoglobin oxygenation in dark-cutting beef. SignificanceThe current understanding indicates that defective glycolysis causes less carbon flow, leading to less postmortem lactic acid formation and elevated muscle pH in dark-cutting beef. However, to the best of our knowledge, limited research has evaluated how changes in glycolytic and mitochondrial protein abundance regulate postmortem muscle acidification and oxygen consumption in dark-cutting beef. We utilized a shotgun proteomics approach to elucidate potential differences in protein profiles between dark-cutting versus normal-pH beef that may influence differences in postmortem metabolism and muscle surface color characteristics. Our study shows that down-regulation of glycolgenolytic and IMP/AMP biosynthetic proteins results in elevated postmortem muscle pH in dark-cutting beef. In addition, the up-regulation of mitochondrial protein content coupled with the higher muscle pH are conducive factors for enhanced oxygen consumption and less myoglobin oxygenation, contributing to a dark meat color typically associated with dark-cutting beef.

Okra Modulates Regulatory Enzymes and Metabolites of Glucose-Utilizing Pathways in Diabetic Rats

Objective Using a rat diabetes model, the authors examined how substrates and products of glycolysis and key regulatory enzymes for glycolysis, gluconeogenesis, Kreb’s cycle, and glycogen metabolism react to treatment with okra diet therapy, relative to glibenclamide treatment. Method The animal grouping involved normoglycemic rats, untreated diabetic rats, and diabetic rats treated with glibenclamide, 50% w/w okra sauce, exclusive okra sauce diet, or sauce without okra. Alloxan monohydrate was the diabetogenic agent. Insulin and adiponectin were assayed with enzyme-linked immunosorbent assay (ELISA) while the metabolites and enzymes were assed using standard spectrophotometric methods. Results The exclusive diet therapy significantly (p < 0.05) improved insulin activities after 60 days and reversed the altered adiponectin activities. Glucose-6-phosphate, fructose-6-phosphate, and fructose-1,6-bisphosphate levels were depleted during diabetes, but phosphoenolpyruvate and pyruvate accumulated during the first short phase of diabetes. Rats in the glibenclamide and 100% okra diet groups showed comparable hexokinase, phosphofructokinase, and pyruvate kinase activities relative to the normoglycemic rats, while the gluconeogenic enzymes, glucose-6-phosphatase, and fructose-1,6-bisphosphatase remained altered. The authors observed that extended treatment with glibenclamide restored the activities of all the Kreb’s cycle enzymes, while succinate dehydrogenase and α-ketoglutarate dehydrogenase were nonresponsive to the okra diet therapy relative to their control levels. The glycogen stores were normalized by the exclusive diet therapy, but glycogen synthase and phosphorylase activities were unresponsive. Conclusions Okra diet has shown insulin-sensitizing potentials with prolonged intake during diabetes as well as the potential to reverse alterations in the major carbohydrate-metabolizing enzyme.

Myosin heavy chain isoform and metabolic profile differ in beef steaks varying in tenderness

Our objective was to investigate possible differences in muscle fiber characteristics of beef longissimus lumborum (LL) steaks varying in tenderness (very tender vs. intermediate tender). Therefore, the relative abundance of myosin heavy chain (MHC) isoforms and activity/abundance of several glycolytic and oxidative enzymes were compared between the two steak groups. Greater (P < 0.05) content of MHC type IIa (MHC-IIa) and activities of phosphofructokinase (PFK) and glycogen phosphorylase (GP) were observed in the very tender steaks. Conversely, intermediate tender steaks had greater (P < 0.05) contents of MHC type I (MHC-I) and succinate dehydrogenase (SDH) and greater citrate synthase (CS) activity. Increased tenderness in the very tender steaks was associated with greater (P < 0.05) proteolysis as evaluated by desmin and troponin-T degradation. Further, mitochondrial calcium uniporter (MCU) was lower (P < 0.05) in the very tender steaks than steaks of intermediate tenderness. Collectively, shifting muscle characteristics toward a more glycolytic type appears to positively impact postmortem proteolysis and tenderization.

Antidiabetic Potency, Antioxidant Effects, and Mode of Actions of Citrus reticulata Fruit Peel Hydroethanolic Extract, Hesperidin, and Quercetin in Nicotinamide/Streptozotocin-Induced Wistar Diabetic Rats.

This study is aimed at assessing the antihyperglycemic, antihyperlipidemic, and antioxidant effects of Citrus reticulata (C. reticulata) fruit peel hydroethanolic extract and two flavonoids, hesperidin and quercetin, in nicotinamide (NA)/streptozotocin- (STZ-) induced type 2 diabetic rats. In addition, GC-MS and HPLC-MS analyses of the extract were performed and the results indicated the presence of multiple flavonoids including hesperidin, quercetin, naringin, and polymethoxylated flavones (nobiletin and tangeretin). To achieve the aim of the study, diabetic rats with NA/STZ-induced T2DM were orally treated with C. reticulata fruit peel hydroethanolic extract, hesperidin, and quercetin at a dose of 100 mg/kg b.w./day for four weeks. The treatments with C. reticulata fruit peel extract, hesperidin, and quercetin significantly ameliorated the impaired oral glucose tolerance; the elevated serum fructosamine level; the diminished serum insulin and C-peptide levels; the altered HOMA-IR, HOMA-IS, and HOMA-β cell function; the decreased liver glycogen content; the increased liver glucose-6-phosphatase and glycogen phosphorylase activities; the deleteriously affected serum lipid profile; the elevated serum AST and ALT activities; and the raised serum creatinine and urea levels in the diabetic rats. The treatments also produced remarkable improvement in the antioxidant defense system manifested by a decrease in the elevated liver lipid peroxidation and an increase in the lowered glutathione content and GPx, GST, and SOD activities. Furthermore, the three treatments enhanced the mRNA expression of GLUT-4 and the insulin receptor β-subunit, but only quercetin produced a significant increase in the expression of adiponectin in adipose tissue of diabetic rats. In conclusion, C. reticulata fruit peel hydroethanolic extract, hesperidin, and quercetin have potent antidiabetic effects which may be mediated through their insulinotropic effects and insulin-sensitizing actions. In addition, the alleviation of the antioxidant defense system by the extract, hesperidin, and naringin may have an important action to enhance the antidiabetic actions and to improve liver and kidney functions in NA/STZ-induced diabetic rats.

Caffeic Acid Protects against Iron-Induced Cardiotoxicity by Suppressing Angiotensin-Converting Enzyme Activity and Modulating Lipid Spectrum, Gluconeogenesis and Nucleotide Hydrolyzing Enzyme Activities.

The protective effectsof caffeic acid on angiotensin-converting enzyme (ACE) and purinergic enzyme activities, as well as gluconeogenesis was investigated in iron-induced cardiotoxicity. Cardiotoxicity was induced in heart tissues harvested from healthy male SD rats by 0.1mM FeSO4. Treatment was carried out by co-incubating hearts tissues with caffeic acid and 0.1mM FeSO4. Cardiotoxicity induction significantly (p < 0.05) depleted GSH level, SOD, catalase, and ENTPDase activities, with concomitant elevation of the levels of malondialdehyde (MDA), nitric oxide, ACE, ATPase, glycogen phosphorylase, glucose 6-phosphatase, fructose 6-biphsophatase, and lipase activities. There was significant (p < 0.05) reversion in these levels and activities on treatment with caffeic acid. Caffeic acid also caused depletion in cardiac levels of cholesterol, triglyceride, LDL-c, while elevating HDL-c level. Our results suggest the protective effect of caffeic acid against iron-mediated cardiotoxicity as indicated by its ability to suppress oxidative imbalance and ACE activity, while concomitantly modulating nucleotide hydrolysis and metabolic switch.

Effects of different ATP contents on phosphorylation level of glycogen phosphorylase and its activity in lamb during incubation at 4 ℃ in vitro

AbstractThe aim of this study was to investigate the effects of ATP on glycogen phosphorylase activity in lamb during incubation at 4℃ in vitro. Sarcoplasmic proteins from lamb were extracted and treated with different contents of ATP to get three groups of glycogen phosphorylase with low, middle and high ATP content groups, the amount of ATP were 0.5, 2.0 and 2.5 μM per 100 μg protein, respectively. The control group was without ATP adding. The results showed that ATP promoted the phosphorylation of glycogen phosphorylase, and phosphorylation modification promoted its activity. But ATP inhibited the activity of glycogen phosphorylase and ATP preferentially participated in phosphorylation. When ATP concentration was 0.5 μM per 100 μg protein, the effect of phosphorylation modification on the activity of glycogen phosphorylase was equal to the inhibition of ATP. The effect of glycogen phosphorylase phosphorylation on its activity gradually became dominant as incubation time prolonged.

Dacryodes edulis (G. Don) H.J. Lam modulates glucose metabolism, cholinergic activities and Nrf2 expression, while suppressing oxidative stress and dyslipidemia in diabetic rats

Ethnopharmacological relevanceDacryodes edulis L. is an evergreen tree indigenous to western and eastern Africa which is utilized for nutritional and medicinal purposes. Folklorically, different parts of the tree are used in treating and managing diabetes and its complications. AimsThe antidiabetic effect of the butanol fraction of D. edulis ethanol extract (BFDE) was studied in fructose-streptozotocin induced type 2 diabetic rats. MethodsThe ethanol extract was fractionated to yield the hexane, dichloromethane, ethyl acetate, butanol and aqueous fractions. The in vitro antidiabetic activities of the fractions were determined by their ability to inhibit α-glucosidase activity. BDFE was the most active and showed no cytotoxic effect while stimulating glucose uptake in 3T3-L1 adipocytes. Thus, selected for in vivo study. Diabetic rats were grouped into 4. The negative control group was administered water only, another group was treated with metformin (200 mg/kg bodyweight), while the other groups were administered BDFE at 150 and 300 mg/kg bodyweight respectively. Two other groups consisting of normal rats were given water and BFDE (300 mg/kg bodyweight) respectively, with the former serving as normal control. After 6 weeks of intervention, the rats were humanely sacrificed using appropriate anaesthesia. ResultsTreatment with the fraction significantly (p < 0.05) reduced the blood glucose level of the diabetic rats, with concomitant increase in serum insulin secretion. It also caused significant (p < 0.05) elevation of reduced glutathione level, superoxide dismutase, catalase, α-amylase, and ATPase activities, with concomitant depletion in myeloperoxidase activity, NO and MDA levels of the serum and pancreas. The pancreatic morphology and β-cell function were significantly improved in BFDE-treated rats, with restoration of the pancreatic capillary networks. Treatment with BFDE significantly (p < 0.05) inhibited the activities of glycogen phosphorylase, fructose 1,6 biphosphatase, glucose 6 phosphatase, and acetylcholinesterase, while suppressing the expression of Nrf2. HPLC analysis revealed the presence of gallic acid, vanillic acid, vanillin, and (−)-epicatechin in the fraction. ConclusionThese results portray the antidiabetic and antioxidative properties of BFDE, which may be a synergistic consequence of the identified phenolics.

Role of phosphorylation on characteristics of glycogen phosphorylase in lamb with different glycolytic rates post-mortem

The relationship between glycogen phosphorylase activity and phosphorylation levels in the longissimus thoracis muscle post-mortem was studied. Sixty lamb samples were collected at 0.5 h, 2 h, 6 h, 12 h, 24 h, 48 h, and 72 h post-mortem and divided into three groups (n = 6) with different glycolytic rates (fast, intermediate, and slow) according to the pH at 6 h post-mortem. The phosphorylation level and activity and expression of glycogen phosphorylase were determined. The results showed that the phosphorylation level and activity of glycogen phosphorylase in the slow pH decline group was lower than that in the fast pH decline group during 24 h post-mortem (P < .05). There was a significant positive correlation between the glycogen phosphorylase activity and the phosphorylation level. In conclusion, these data demonstrated that the glycogen phosphorylase activity in lambs was affected by phosphorylation levels and postmortem duration.

Effects of low salinity on gill and liver glycogen metabolism of great blue-spotted mudskippers (Boleophthalmus pectinirostris)

This study investigated the effects of low salinity exposure on glycogen and its metabolism biomarkers, glycogen synthase (GS) and glycogen phosphorylase (GP), representing glycogen synthesis and catabolism, respectively, in the gills and liver of great blue-spotted mudskippers (Boleophthalmus pectinirostris). The fish were accumulated at 10‰ salinity seawater for 1 week, then 270 healthy great blue-spotted mudskippers with similar size were randomly transferred to 10‰ (control group) or 3‰ (low salinity group) seawater for 72-hour stress experiment. Fish significantly elevated their blood glucose levels 12 h after low salinity challenge. At the end of experiments, a decrease in liver glycogen contents was observed in both the control and low salinity groups, the latter showing a pronounced decrease, while the gill glycogen contents were not changed for either group. The mRNA abundance and enzyme activity of GS and GP were both elevated in gill tissues, showing a rising glycogen synthesis and catabolism, probably resulting in the unchanging gill glycogen content. While in liver tissues, the mRNA abundance and enzyme activity were decreased for GS and increased for GP, showing a net increase for breaking down glycogen in liver, probably for supplying a sufficient glucose level for gills and other tissues/organs involved in the response to salinity changes.

Comparison of activity, expression and S-nitrosylation of glycolytic enzymes between pale, soft and exudative and red, firm and non-exudative pork during post-mortem aging

The activity, expression and S-nitrosylation of glycogen phosphorylase (GP), phosphofructokinase (PFK) and pyruvate kinase (PK) was compared between pale, soft and exudative (PSE) and red, firm and non-exudative (RFN) pork. The nitric oxide synthase (NOS) activity of RFN pork was higher than PSE pork (P < 0.05). Glycogen and lactic acid content were significantly different between PSE and RFN samples at 1 h postmortem (P < 0.05). Compared to PSE pork, RFN pork had lower activities and higher S-nitrosylation levels of GP, PFK and PK (P < 0.05). Moreover, GP expression in RFN pork was lower (P < 0.05) while no significant differences of PFK and PK expression were observed between these two groups. These data suggest that protein S-nitrosylation can presumably regulate glycolysis by modulating glycolytic enzymes activities and then regulate the development of PSE pork.

Effects of phosphorylation on the activity of glycogen phosphorylase in mutton during incubation at 4 °C in vitro

The aim of this study was to assess the effects of the phosphorylation levels of glycogen phosphorylase on its activity in mutton sarcoplasmic protein samples during incubation at 4 °C. Samples of sarcoplasmic proteins from mutton longissimus thoracis muscles were prepared and separated into three treatment groups to obtain glycogen phosphorylase with different phosphorylation levels, which were (1) treated with protein kinase A, (2) treated with alkaline phosphatase, and (3) left untreated (control). Glycogen phosphorylase phosphorylation levels and activity as well as the levels of related endogenous substances were assessed. The results showed that phosphorylation of glycogen phosphorylase in mutton promoted its activity during incubation at 4 °C. The activity of glycogen phosphorylase was also influenced by other factors (glycogen, glucose, glucose 6-phosphate, ATP, etc.) in vitro. The combined effects of phosphorylation and endogenous substances on glycogen phosphorylase activity varied at different incubation times.

The Influence of Mitochondria Enzyme Activity on Beef Tenderness

ObjectivesAmong all eating quality attributes, tenderness is often described as the most important factor dictating the overall acceptance of cooked beef, as well as, future decision to repeat purchase. While many intrinsic and extrinsic factors impact end-product tenderness, variation in tenderness is usually attributed to the extent of postmortem proteolysis occurring during meat aging. Evidence from the literature indicates that muscle fiber type composition is a major source of variation in the rate and extent of postmortem proteolysis. One of the major differences that distinguishes muscle fibers is the content of mitochondria. Typically, red muscle (slow-oxidative) is characterized by greater amounts of mitochondria than white muscle (fast-glycolytic). As part of the calcium buffering system, mitochondria sequester large quantities of calcium to maintain a constant cytosolic calcium level. We hypothesized that mitochondria may delay the activation of µ-calpain, the major protease responsible for postmortem proteolysis, through preventing the increase in cytosolic calcium concentration.Materials and MethodsTo test our hypothesis, beef longissimus thoracis muscle samples were collected at 30 min and 16 d postmortem. The 30 min samples were immediately snap frozen in liquid nitrogen and stored at – 80°C, while the 16 d samples were used to determine Warner-Bratzler shear force (WBSF) values. Based on WBSF values, the samples were allocated into less tender (average WBSF = 5.3 kg; n = 8) or more tender (average WBSF = 2.3 kg; n = 8) categories. Succinate dehydrogenase (SDH) abundance, citrate synthase (CS) activity, phosphofructokinase (PFK) activity, and glycogen phosphorylase (GP) activity were compared between the two categories using the 30 min samples. Collected data were analyzed using a Student’s t test and considered significant at P ≤ 0.05.ResultsOur results showed that SDH abundance and CS activity (mitochondrial biomarkers) were significantly greater (P = 0.01 and 0.003, respectively) in less tender samples when compared to more tender samples. On the other hand, PFK and GP activities (glycolytic biomarkers) were greater (P &lt; 0.05) in the more tender than less tender steaks.ConclusionWhile not a cause and effect relationship, these data indicate that mitochondria content likely plays a role in development of beef tenderness.

McArdle Disease: New Insights into Its Underlying Molecular Mechanisms.

McArdle disease, also known as glycogen storage disease type V (GSDV), is characterized by exercise intolerance, the second wind phenomenon, and high serum creatine kinase activity. Here, we recapitulate PYGM mutations in the population responsible for this disease. Traditionally, McArdle disease has been considered a metabolic myopathy caused by the lack of expression of the muscle isoform of the glycogen phosphorylase (PYGM). However, recent findings challenge this view, since it has been shown that PYGM is present in other tissues than the skeletal muscle. We review the latest studies about the molecular mechanism involved in glycogen phosphorylase activity regulation. Further, we summarize the expression and functional significance of PYGM in other tissues than skeletal muscle both in health and McArdle disease. Furthermore, we examine the different animal models that have served as the knowledge base for better understanding of McArdle disease. Finally, we give an overview of the latest state-of-the-art clinical trials currently being carried out and present an updated view of the current therapies.

Effects of preslaughter shackling on postmortem glycolysis, meat quality, changes of water distribution, and protein structures of broiler breast meat

The objective of this study was to investigate the effects of preslaughter shackling on stress, postmortem glycolysis, meat quality, water distribution, and protein structures of pectoralis majors. Before slaughter, Arbor Acres broilers (n = 105, 42 days old, 2.0 to 2.5kg) were randomly categorized into 3 treatment groups: (I) control group without shackling (NS); (II) 2.5min shackling (SS); (III) 4.5min shackling (LS). Each treatment group consisted of 5 replicates with 7 broilers each. Results indicated that preslaughter shackling increased (P < 0.05) plasma corticosterone and adrenocorticotropic hormone concentrations in comparison with the control group. Antemortem shackling increased (P < 0.05) activity of glycogen phosphorylase and phosphofructokinase-1 (PFK-1) accompanying with rapid glycolysis and pH decline at early postmortem. LS treatment led to myosin denaturation, decreased (P < 0.05) α-helix content, and increased (P < 0.05) β-sheet structures proportion in the myofibrillar proteins. Furthermore, meat from LS treatment had higher (P < 0.05) lightness, redness, and poorer water-holding capacity. These results indicated that the longer shackling duration (4.5min) increased stress and the rate of glycolysis, causing myosin denaturation and changes of the secondary structure in the myofibrillar proteins, which aggravated the deterioration of meat quality.

Hesperidin, a citrus flavonoid ameliorates hyperglycemia by regulating key enzymes of carbohydrate metabolism in streptozotocin-induced diabetic rats

The present study investigated the protective effect of hesperidin on carbohydrate metabolizing enzymes in streptozotocin-induced diabetic rats. Hesperidin was administered to streptozotocin-induced (40 mg/kg b.w.) diabetic rats at different dosages of (25, 50, 100 mg/kg b.w.) respectively for 30 days to evaluate its effect on fasting plasma glucose, insulin, glycosylated hemoglobin, hemoglobin, and carbohydrate metabolic enzymes. The plasma glucose levels were significantly reduced in a dose-dependent manner in hesperdin-treated group of rats when compared to the diabetic control rats. In addition, concomitant increase in hemoglobin and insulin levels and a decrease in glycosylated hemoglobin were observed in treated group of rats. The activities of the hepatic key enzymes of carbohydrate metabolism such as hexokinase and glucose-6-phosphate dehydrogenase were significantly increased, whereas glucose-6-phosphatase and fructose-1,6-bisphosphatase were significantly decreased. Furthermore, hesperidin administration prevented the loss in body weight and improved the glycogen content in the hepatic tissue of diabetic animals by reinstating the activities of glycogen synthase and glycogen phosphorylase. These results showed that hesperidin has potential antihyperglycemic activity in streptozotocin-induced diabetic rats. This was further supported by the histological studies of pancreas and liver.

The Role of Hormones in the Regulation of Glycogen Metabolism in the Clawed Toad Xenopus Laevis (Daudin)

The hormonal regulation of amphibian glycogen metabolism was studied in Xenopus laevis as a typical member of the anurans (tailless amphibians).The main focus of this study was given to the effects of various hormones on the glycogen/glucose balance in adult toads. We determined biochemically the liver and muscle glycogen contents as well as the blood glucose and lipid levels for a number of hormones and also diabetes inducing substances. Additionally, we examined ultrastructure changes in hepatocytes induced by the various treatments, and also investigated the activity of carbohydrate-relevant enzymes by histochemistry. With one exception, the liver glycogen content of Xenopus remained basically unchanged by the treatments or was even slightly enhanced. Only human chorionic gonadotropin, through which the vitellogenic response is triggered, prompts a significant decrease of liver glycogen in females. Under the same conditions the male liver glycogen content remained stable. Muscle glycogen contents were not affected by any of the treatments. Blood glucose and lipid levels on the other hand were elevated considerably in both sexes after application of either epinephrine or cortisol. The ultrastructural examination revealed a proliferation of Rough Endoplasmic Reticulum (RER) in hepatocytes from epinephrine treated toads of both sexes as well as from HCG treated females. By histochemistry, we detected an elevated glucose-6-phosphatase activity in the hepatocytes from toads treated with either epinephrine or cortisol. These treatments also led to enhanced glycogen phosphorylase activity in males, and to a slightly elevated glyceraldehyde-3-phosphate dehydrogenase activity in females. Our results show that the hepatic glycogen is extremely stable in adult Xenopus. Only vitellogenesis causes a marked utilization of glycogen. Since the blood glucose levels are elevated in epinephrine or cortisol treated toads without the liver glycogen being affected, we conclude that either protein and/or lipid metabolism are involved in carbohydrate metabolism in Xenopus laevis.

The antidiabetic properties of the hot water extract of kola nut (Cola nitida (Vent.) Schott & Endl.) in type 2 diabetic rats

Ethnopharmacological relevanceCola nitida is amongst the evergreen plants native to West Africa used in the treatment of various ailments including diabetes.Aim of the study: This study aims to investigate the antidiabetic effects of the hot water extract of C. nitida seeds in type 2 diabetic rats. MethodsType 2 diabetic rats were orally administered with low (150 mg/kg bw) and high (300 mg/kg bw) doses of the hot water extract for 6 wk and thereafter, blood glucose, insulin level, lipid profile, pancreatic β-cell function, perfusion and morphology, redox imbalance, glycolytic and cholinergic enzymes, as well as of caspase-3 and Nrf2 expressions were measured. ResultsTreatment with the extract led to significant depletion of blood glucose, serum triglycerides, LDL-cholesterol, fructosamine, ALT, and uric acids, while elevating serum insulin and HDL-cholesterol levels. The infusion also significantly (p < 0.05) elevated GSH level, SOD, catalase, α-amylase, and ATPase activities, with concomitant depletion of myeloperoxidase enzyme activity, and NO and MDA levels in the serum and pancreas. Significantly (p < 0.05) improved pancreatic β-cell function and morphology were observed in rats treated with C. nitida, with restored pancreatic capillary networks. C. nitida inhibited the activities of glycogen phosphorylase, fructose 1,6 biphosphatase, glucose 6 phosphatase, and acetylcholinesterase while downregulated the Nrf2 expression. NMR analysis of the extract revealed the presence of caffeine and theobromine. The molecular docking studies indicated that identified compounds displayed strong molecular interactions with caspase-3 and Nrf2. ConclusionThese results insinuate the antidiabetic activities of C. nitida hot water extract and may be attributed to the NMR-identified compounds.

Effects of protein phosphorylation on glycolysis through the regulation of enzyme activity in ovine muscle

To verify the effect of protein phosphorylation on glycolysis and elucidate the regulatory mechanism from the perspective of enzyme activity, ovine muscle was treated with a kinase inhibitor, dimethyl sulfoxide, or a phosphatase inhibitor and the activities of glycogen phosphorylase, pyruvate kinase and phosphofructokinase were determined. The protein phosphorylation level was significantly different after incubation of muscle with kinase or phosphatase inhibitors. The pH value and lactate content revealed that a high phosphorylation level was the reason for the fast glycolysis. The glycogen phosphorylase, pyruvate kinase and phosphofructokinase activities were significantly higher in the phosphatase inhibitor group than in the other two groups (p < 0.05). Therefore, protein phosphorylation is involved in activating these three enzymes. In summary, protein phosphorylation plays a role in post-mortem glycolysis through the regulation of enzyme activity in ovine muscle.

Low-level free nitrous acid efficiently inhibits the conjugative transfer of antibiotic resistance by altering intracellular ions and disabling transfer apparatus

Recently, the dissemination of antibiotic resistance genes (ARGs) via plasmid-mediated conjugation has been reported to be facilitated by a series of contaminants. This has highlighted potential challenges to the effective control of this principal mode of horizontal transfer. In the present study, we found that low levels (<0.02 mgN/L) of free nitrous acid (FNA) remarkably inhibited (over 90%) the conjugative transfer of plasmid RP4, a model broad-host-range plasmid, between Escherichia coli. The antimicrobial role of FNA at the applied dosages was firstly ruled out, since no dramatic reductions in viabilities of donor or recipient were observed. Instead, FNA appeared to reduce the available intracellular free Mg2+, which was confirmed to be triggered by the liberation of intracellular Fe2+. These alterations in intracellular Mg2+ and Fe2+ concentrations were found to significantly limit the available energy for conjugative transfer through suppression of glycolysis by decreasing the activities of glycogen phosphorylase and glyceraldehyde-3-phosphate dehydrogenase and also by diverting the glycolytic flux into the pentose phosphate pathway via activation of glucose-6-phosphate dehydrogenase towards the generation of NADPH rather than ATP. Moreover, RP4-encoding genes responsible for DNA transfer and replication (traI, traJ and trfAp), coupling (traG) and mating pair formation (traF and trbBp) were all significantly down-regulated after FNA treatment, indicating that the transfer apparatus required for plasmid processing and delivery was deactivated. By validating the inhibitory effects of FNA on conjugation in real wastewater, this study highlights a promising method for controlling the dissemination of ARGs in systems such as wastewater treatment plants.