Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) via dephosphorylation of the EGFR signaling pathway

The current study aimed to assess the effect of timosaponin AⅢ(T-AⅢ)on drug-metabolizing enzymes during anticancer therapy.The in vivo experiments were conducted on nude and ICR mice.Following a 24-day administration of T-AⅢ,the nude mice exhibited an induction of CYP2B10,MDR1,and CYP3A11 expression in the liver tissues.In the ICR mice,the expression levels of CYP2B10 and MDR1 increased after a three-day T-AⅢ administration.The in vitro assessments with HepG2 cells revealed that T-AⅢ induced the expression of CYP2B6,MDR1,and CYP3A4,along with constitutive androstane receptor(CAR)activation.Treatment with CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4 expression.Furthermore,other CAR target genes also showed a significant increase in the expression.The up-regulation of murine CAR was observed in the liver tissues of both nude and ICR mice.Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation,with this effect being partially reversed by the ERK activator t-BHQ.Inhibition of the ERK1/2 signaling pathway was also observed in vivo.Additionally,T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845,and suppressed EGF-induced phosphorylation of EGFR,ERK,and CAR.In the nude mice,T-AⅢ also inhibited EGFR phosphorylation.These results collectively indicate that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.

Processing Effects on Selected Antioxidant Activities and Metabolizing Enzyme Inhibition of M. Koneigii (Curry Leaves) Extracts

Curry leaves, scientifically termed Murraya koenigii, are renowned in South Asian cuisine for their flavor enhancement and potential health benefits, including antioxidative, anti-inflammatory, and antidiabetic properties. This study aimed to evaluate the impact of thermal processing methods on curry leaves by analysing Total Phenolic Content (TPC), Total Flavonoid Content (TFC), antioxidant activity, and metabolizing enzyme inhibition. Fresh curry leaves were subjected to thermal treatments: Oven-dried at 60˚C and Air-dried at 25˚C for 2 weeks. Extracts were prepared using Ethanol and water solvents. Results indicated that Air-dried leaves exhibited significantly higher TPC (5132.65 mg GAE/100 g) and TFC (243.13 mg CE/100 g) compared to Fresh and Oven-dried leaves. Antioxidant assays show that oven-dried curry leaves at 60˚C displayed higher results in NORS, FRAP, and TEAC assays compared to Fresh and Air-dried leaves. Ethanol extracts showed better extraction of bioactive compounds than aqueous extracts. Moreover, Lipase inhibition activity was notably high, indicating potential health benefits. This study provides valuable insights into the effects of processing methods on curry leaf extracts, emphasizing the importance of solvent selection for optimal extraction of bioactive compounds.

Salvianolic acid B modulates the expression of drug-metabolizing enzymes in HepG2 cells

BACKGROUND: Enzymes involved in drug and xenobiotic metabolism have been considered to exist in two groups: phase I and phase II enzymes. Cytochrome P450 isoenzymes (CYPs) are the most important phase I enzymes in the metabolism of xenobiotics. The products of phase I metabolism are then acted upon by phase II enzymes, including glutathione S-transferases (GSTs). Herbs that inhibit CYPs such as CYP3A4 or that induce GSTs may have the potential to protect against chemical carcinogenesis since the mutagenic effects of carcinogens are often mediated through an excess of CYP-generated reactive intermediates. This study was designed to investigate the effects of salvianolic acid B (Sal B), a pure compound extracted from Radix Salviae Miltiorrhizae, a Chinese herb, on cell proliferation and CYP1A2 and CYP3A4 mRNA expression in the presence or absence of rifampicin, a potent inducer of CYPs and GST protein expression in HepG2 cells. METHODS: HepG2 cells were incubated with different concentrations of Sal B. Cell proliferation was determined by SYTOX-Green nucleic acid staining. CYP3A4 and CYP1A2 mRNA expression was assayed by real-time PCR. GST protein expression was analyzed by Western blotting. RESULTS: Low concentrations of Sal B (0-20 μmol/L) had no significant effects on cell proliferation, while higher concentrations (100-250 μmol/L) significantly inhibited proliferation in a concentration-dependent manner. Ten μmol/L Sal B, but not 1 μmol/L, down-regulated CYP3A4 and CYP1A2 mRNA expression after 24 hours of incubation, whereas both 1 and 10 μmol/L Sal B down-regulated CYP3A4mRNA expression after 96 hours of incubation; moreover, 1 and 10 μmol/L Sal B inhibited CYP3A4 mRNA expression induced by rifampicin. Both 1 μmol/L and 10 μmol/L Sal B increased GST expression. CONCLUSION: Sal B inhibits CYP3A4 and CYP1A2 mRNA expression and induces GST expression in HepG2 cells.

Hepatic angiotensin-converting enzyme 2 expression in metabolic dysfunction-associated steatotic liver disease and in patients with fatal COVID-19

BACKGROUND Metabolic dysfunction-associated steatotic liver disease(MASLD),characterised by hepatic lipid accumulation,causes inflammation and oxidative stress accompanied by cell damage and fibrosis.Liver injury(LI)is also frequently reported in patients hospitalised with coronavirus disease 2019(COVID-19),while preexisting MASLD increases the risk of LI and the development of COVID-19-associated cholangiopathy.Mechanisms of injury at the cellular level remain unclear,but it may be significant that severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)which causes COVID-19,uses angiotensin-converting expression enzyme 2(ACE2),a key regulator of the‘anti-inflammatory’arm of the renin-angiotensin system,for viral attachment and host cell invasion.AIM To determine if hepatic ACE2 levels are altered during progression of MASLD and in patients who died with severe COVID-19.METHODS ACE2 protein levels and localisation,and histological fibrosis and lipid droplet accumulation as markers of MASLD were determined in formalin-fixed liver tissue sections across the MASLD pathological spectrum(isolated hepatocellular steatosis,metabolic dysfunction-associated steatohepatitis(MASH)+/-fibrosis,end-stage cirrhosis)and in post-mortem tissues from patients who had died with severe COVID-19,using ACE2 immunohistochemistry and haematoxylin and eosin and picrosirius red staining of total collagen and lipid droplet areas,followed by quantification using machine learning-based image pixel classifiers.RESULTS ACE2 staining is primarily intracellular and concentrated in the cytoplasm of centrilobular hepatocytes and apical membranes of bile duct cholangiocytes.Strikingly,ACE2 protein levels are elevated in non-fibrotic MASH compared to healthy controls but not in the progression to MASH with fibrosis and in cirrhosis.ACE2 protein levels and histological fibrosis are not associated,but ACE2 and liver lipid droplet content are significantly correlated across the MASLD spectrum.Hepatic ACE2 levels are also increased in COVID-19 patients,especially those showing evidence of LI,but are not correlated with the presence of SARS-CoV-2 virus in the liver.However,there is a clear association between the hepatic lipid droplet content and the presence of the virus,suggesting a possible functional link.CONCLUSION Hepatic ACE2 levels were elevated in nonfibrotic MASH and COVID-19 patients with LI,while lipid accumulation may promote intra-hepatic SARS-CoV-2 replication,accelerating MASLD progression and COVID-19-mediated liver damage.

Addition of Protease Enzyme to Dog and Cat Feed and Its Influence on the Digestibility Coefficient, Immune Response, and Metabolic Biomarkers

Promoting better protein digestibility through exogenous enzymes in the diet is essential in the nutrition of companion animals, mainly for better performance and maintenance of the animals’ physiological and metabolic systems, promoting health and adequate growth. In addition, it reduces the cost of the diet by possibly reducing protein in the diet, which is the main and most expensive ingredient for dogs and cats. The objective of this study was to verify whether the addition of protease to dog and cat food can improve the protein digestibility of the food, thus facilitating greater absorption of amino acids and influencing metabolic biomarkers and immune response. To this end, two experiments were carried out to evaluate the protease from the fermentation of Aspergillus niger and Bacillus subtilis. Experiment 1 was carried out with ten male, non-castrated beagles divided into two groups of five animals: the control group (without enzyme) and the test group (with 250 g of protease/ton). The animals underwent two 45-day experimental periods, and in the second period, after a 15-day interval, the dogs in the control group became part of the treatment group (crossover model). Adding this enzyme to the dogs’ diet had no adverse effects on the animals’ health besides improving the digestibility of dry matter and crude protein consumed by the dogs. Experiment 2 was carried out with sixteen female cats of no defined breed, non-castrated, divided into four groups with four animals per group, namely: Treatment A (without enzyme), Treatment B (with protease at a dose of 100 g/ton), Treatment C (with protease at a dose of 200 g/ton) and Treatment D (with protease at a dose of 400 g/ton). The cats underwent two 30-day experimental periods, and in the second period, after a 15-day interval, the animals switched between treatments (crossover model) to increase the power of the statistical test. The enzyme consumption did not affect the felines’ metabolism and health but improved the digestibility of crude protein at doses of 200 and 400 g/ton. The results allow us to conclude that the protease used in this study can improve the digestibility of crude protein for dogs and cats.

Screening for metabolic dysfunction-associated fatty liver disease:Time to discard the emperor’s clothes of normal liver enzymes?

Metabolic dysfunction-associated fatty liver disease(MAFLD)is the most prevalent chronic liver condition worldwide.Current liver enzyme-based screening methods have limitations that may missed diagnoses and treatment delays.Regarding Chen et al,the risk of developing MAFLD remains elevated even when alanine aminotransferase levels fall within the normal range.Therefore,there is an urgent need for advanced diagnostic techniques and updated algorithms to enhance the accuracy of MAFLD diagnosis and enable early intervention.This paper proposes two potential screening methods for identifying individuals who may be at risk of developing MAFLD:Lowering these thresholds and promoting the use of noninvasive liver fibrosis scores.

Effects of frying oil and Houttuynia cordata thunb on xenobiotic-metabolizing enzyme system of rodents

AIM: To evaluate the effects of frying oil and Houttuynia cordata Thunb (H. cordata), a vegetable traditionally consumed in Taiwan, on the xenobiotic-metabolizing enzyme system of rodents. METHODS: Forty-eight Sprague-Dawley rats were fed with a diet containing 0%, 2% or 5% H. cordata powder and 15% fresh soybean oil or 24-h oxidized frying oil (OFO) for 28 d respectively. The level of microsomal protein, total cytochrome 450 content (CYP450) and enzyme activities including NADPH reductase, ethoxyresorufin 0-deethylase (EROD), pentoxyresorufin 0-dealkylase (PROD), aniline hydroxylase (ANH), aminopyrine demethylase (AMD), and quinone reductase (QR) were determined. QR represented phase Ⅱ enzymes, the rest of the enzymes tested represented phase Ⅰ enzymes. RESULTS: The oxidized frying oil feeding produced a significant increase in phase Ⅰ and Ⅱ enzyme systems, including the content of CYP450 and microsomal protein, and the activities of NADPH reductase, EROD, PROD, ANH, AMD and QR in rats (P<0.05). In addition, the activities of EROD, ANH and AMD decreased and QR increased after feeding with H. cordata in OFO-fed group (P<0.05). The feeding with 2% H. cordata diet showed the most significant effect. CONCLUSION: The OFO diet induces phases I and II enzyme activity, and the 2% H. cordata diet resulted in a better regulation of the xenobiotic-metabolizing enzyme system.

Changes in mRNA Expression and Activity of Xenobiotic Metabolizing Enzymes in Livers from Adjuvant-Induced Arthritis Rats

Pathophysiological changes in human patients and in animal models of infection or inflammation are associated with alterations in the production of numerous liver-derived proteins including metabolizing enzymes. In this study, the effects of adjuvant-induced arthritis (AA) in rats on the levels of mRNA and activity of hepatic xenobiotic metabolizing enzymes were determined during the inflammatory response. The mRNA levels of cytochrome P450 (CYP) 1A2, CYP2C12, CYP2D1, CYP2D2, and CYP3A1 were significantly decreased compared with control levels in almost all phases of inflammation. A reduction in the activity of CYP2C and CYP3A, which are abundantly expressed in the liver, was also observed. For phase II metabolizing enzymes, mRNA levels of uridine 5’-diphospho-glucuronosyltransferase (UGT) 1A1, UGT1A6, sulfotransferase (SULT) 2A1, and glutathione S-transferase 2 were significantly decreased compared with control levels. However, the mRNA levels of UGT2B and SULT1A1 returned to control levels during the subacute (7 d after adjuvant treatment) and chronic (21 d after adjuvant treatment) phases although these levels decreased during the acute (3 d after adjuvant treatment) phase. These results suggest that the effects of inflammation on the expression of xenobiotic metabolizing enzymes differ depending on the isoform of the enzyme and could affect the pharmacokinetics of each substrate.

High Temperature at Grain-filling Stage Affects Nitrogen Metabolism Enzyme Activities in Grains and Grain Nutritional Quality in Rice

Rice plants would more frequently suffer from high temperature (HT) stress at the grain-filling stage in future. A japonica rice variety Koshihikari and an indica rice variety IR72 were used to study the effect of high temperature on dynamic changes of glutamine synthetase (GS) activity, glutamate synthase (GOGAT) activity, glutamic oxalo-acetic transminase (GOT) activity, glutamate pyruvate transminase (GPT) activity in grains and grain nutritional quality at the grain-filling stage. Under HT, the activities of GOGAT, GOT, GPT and soluble protein content in grains significantly increased, whereas GS activity significantly decreased at the grain-filling stage. In addition to the increase of protein and amino acids contents, it was suggested that GOGAT, GOT and GPT in grains played important roles in nitrogen metabolism at the grain-filling stage. Since the decrease of GS activity in grains did not influence the accumulations of amino acids and protein, it is implied that GS might not be the key enzyme in regulating glutamine content in grains.

Changes of Reactive Oxygen Species and Related Enzymes in Mitochondria Respiratory Metabolism During the Ripening of Peach Fruit

The fruits of peach cultivar Yuhua 3 were used as materials to investigate the changes of active oxygen and related enzymes in mitochondria respiratory metabolism during ripening of peach fruit, involving their influence on the proceeding of peach fruit senescence. The results showed that the large decrease in firmness occurred between maturity II and IV. The decrease in firmness coincided with an increase in respiratory intensity. Obvious peaks of respiratory intensity lagging to the rapid change of fruit firmness could be shown during peach ripening. Reactive oxygen species （ROS） had a cumulative process and positively correlated with respiratory intensity. During peach ripening, the content of Ca^2＋ increased, the activities of succinic dehydrogenase （SDH）, cytochrome C oxidase （CCO）, H＋-ATPase, and Ca^2＋-ATPase decreased varying in different degree at the later step of ripening. These suggested a close relationship existed between ROS metabolism and mitochondrial respiration, namely, both ROS metabolism and mitochondrial respiration probably played important roles in ripening and senescing of peach fruit.

Gene Expression and Activity of Enzymes Involved in Sugar Metabolism and Accumulation During “Huangguan” and “Yali” Pear Fruit Development

Since the carbohydrate content affects pear flavor during the process of growth, it is necessary to determine the sugar components that accumulate in the fruit. We analyzed the fruit carbohydrate content, and the gene expression and activity ofacid invertase(AI), neutral invertase(NI), sucrose synthase(SS), and sucrose phosphate synthase(SPS) during the development of "Huangguan" and "Yali" pears. The results demonstrate that during development, the fruit sugar metabolism of the "Huangguan" pear follows a typical sorbitol–starch-soluble sugars middle model, whereas the "Yali" pear fruit follows a typical sorbitol–sucrose–starch-soluble sugars middle model. In the "Huangguan" pear, we found the AI and NI gene expressions, as well as AI( P < 0.05) and NI( P < 0.01) enzyme activities, to be positively correlated, whereas we found the NI gene expression and NI enzyme activity of "Yali" pear to be negatively correlated( P < 0.01). We observed the high levels oflate-stage AI and early-stage SS during development to roughly correspond with the gene expression found in the late and early stages, respectively, suggesting their potential regulatory roles in "Huangguan" pear fruit development. Our results indicate that the primary function of SPS during the early developmental stage is to accumulate sucrose, whereas the primary function of AI is to promote hexose accumulation during the late developmental stage ofmature "Yali" pear fruit.

Effects of Increased Night Temperature on Cellulose Synthesis and the Activity of Sucrose Metabolism Enzymes in Cotton Fiber

Temperature is one of the key factors that influence cotton fiber synthesis at the late growth stage of cotton. In this paper, using two early-maturing cotton varieties as experimental materials, night temperature increase was stimulated in the field using far-infrared quartz tubes set in semi-mobile incubators and compared with the normal night temperatures （control） in order to investigate the effects of night temperature on the cotton fiber cellulose synthesis during secondary wall thickening. The results showed that the activity of sucrose synthase （SuSy） and sucrose phosphate synthase （SPS） quickly increased and remained constant during the development of cotton fiber, while the activity of acid invertase （AI） and alkaline invertase （NI） decreased, increased night temperatures prompted the rapid transformation of sugar, and all the available sucrose fully converted into cellulose. With night temperature increasing treatment, an increase in SuSy activity and concentration of sucrose indicate more sucrose converted into UDPG （uridin diphosphate-glucose） during the early and late stages of cotton fiber development. Furthermore, SPS activity and the increased concentration of fructose accelerated fructose degradation and reduced the inhibition of fructose to SuSy; maintaining higher value of allocation proportion of invertase and sucrose during the early development stages of cotton fiber, which was propitious to supply a greater carbon source and energy for cellulose synthesis. Therefore, the minimum temperature in the nightime was a major factor correlated with the activity of sucrose metabolism enzymes in cotton fiber. Consequently, soluble sugar transformation and cellulose accumulation were closely associated with the minimum night temperature.

Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase (PPO) in Mythimna separta Walker

To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase （GSTs）, cytochrome P450 （P450）, and polyphenol oxidase （PPO） of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase （ACP） and alkaline phosphatase （AKP） in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management.

Chrysoeriol ameliorates hyperglycemia by regulating the carbohydrate metabolic enzymes in streptozotocin-induced diabetic rats

The present study aimed to evaluate the effects of chrysoeriol from Cardiospermum halicacabum in streptozotocin induced Wistar rats.Thirty rats were categorized as control,diabetic control supplemented with 0,20 mg/kg chrysoeriol and 600μg/kg BW of glibenclamide for 45-day trial period.Our results indicated that the inclusion of chrysoeriol(20 mg/kg)showed a significant reduction in plasma glucose,hemoglobin and glycosylated hemoglobin level with a rising of plasma insulin sensitivity.Further,downregulated enzymes including glucose 6-phosphatase,fructose 1,6-bisphosphatase,and glycogen phosphorylase as well upregulated enzymes such as hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,and hepatic glycogen content.There was a diminish action found in liver glycogen synthase of tested rat with a rise in gamma-glutamyl transpeptidase,towards normal levels upon treatment with chrysoeriol.The histopathological study confirmed that renewal of the beta cells of pancreatic of chrysoeriol and glibenclamide treated rats.In addition,the molecular docking of chrysoeriol against glycolytic enzymes including hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,using Argus software shows chrysoeriol had greatest ligand binding energy as equivalent to glibenclamide,as a standard drug.Thus,chrysoeriol found to be non-toxic with potential regulation on glycemic control and upregulation of the carbohydrate metabolic enzymes.

Recent Advances on Key Enzyme Genes in Metabolic Pathways Related to the Synthesis of Aroma Compounds in Tobacco

This article summarized three main kinds of metabolic pathways related to the synthesis of aroma compounds in plants, concluded the roles and expres- sion patterns of key enzyme genes catalyzing the formation of major intermediate products in phenylpropanoid metabolic pathway, isoprene metabolic pathway and alkaloid biosynthetic pathway respectively, highlighted the latest developments of these key enzyme genes in tobacco, and accordingly proposed that in-depth study at the protein level and analysis of metabolic network interaction should be carried out in tobacco besides the expression regulation and transgenic crop improvement at the genetic level. Based on the above analysis, further improvement of tobacco aroma quality through metabolic engineering and its application prospect in agricultural production were prospected.

Effects of Drought Stress on Key Enzymes of Carbon Metabolism, Photosynthetic Characteristics and Agronomic Traits of Soybean at the Flowering Stage under Different Soil Substrates

Soybean is an important legume food crop,and its seeds are rich in nutrients,providing humans and animals with edible oil and protein feed.However,soybean is sensitive to water requirements,and drought is an important factor limiting soybean yield and quality.This study used Heinong 84(drought resistant variety)and Hefeng 46(intermediate variety)as tested varieties planted in chernozem,albic,and black soils.The effects of drought stress on the activities of key enzymes in carbon metabolism and photosynthetic characteristics of soybean were studied during the flowering stage,most sensitive to water.(1)The activities of SS-1,6PGDH,and G6PDH enzymes in soybean leaves first increased and then decreased under drought stress.The enzyme activity was the highest under moderate drought stress and weakest in the blank group.(2)Drought stress increased Phi2,PhiNO,and Fm in soybean leaves and reached the highest value under severe drought;with the increase in drought stress,PhiNPQ and Fv/Fm of soybean leaves gradually decreased,reaching the lowest under severe drought.(3)With the increase in drought stress,F0 and Fs of soybean leaves showed a single peak curve,and the maximum was at moderate drought.(4)Correlation analysis showed that F0 was greatly affected by varieties and soil types;Fs,F0,and Fm soil varieties had a great influence,and chlorophyll fluorescence parameters were affected differently under drought stress with different drought degrees.(5)Drought stress changed the agronomic traits and yield of soybean.With the increase of drought degree,plant height,node number of main stem,effective pod number,100-seed weight and total yield decreased continuously.(6)Drought stress affected the dry matter accumulation of soybean.With the increase of drought degree,the dry matter accumulation gradually decreased.Among them,the leaf was most seriously affected by drought,and SD decreased by about 55%compared with CK.Under the condition of black soil,the dry matter accumulation of soybean was least affected by drought.

Effects of Grain Protein Content Selection on Protein Content and Key Enzyme Activity Involved in Nitrogen Metabolism in Progenies Derived from a Rice Cross

Two japonica rice parents （Tong 769 and Xixuan 1） and their progenies, significantly different in protein content of grains, were investigated to reveal the activities of proteinase in leaves and glutamine synthetase in grains, as well as the dynamic changes of soluble protein content in grains during rice grain filling. The results showed that the parents were very similar in protein content, however, advanced lines with different protein contents in grains and varied activities of proteinase and glutamine synthetase were acquired by consecutively directional selection of the grain protein content in their progenies. Moreover, the enzyme activity and the protein content in grains exceeded their parents during grain filling. The protein content in grains was positively related with the proteinase activity, and the soluble protein content was negatively related with the glutamine synthetase activity in grains to some extent.

Proteolytic Enzyme Combination Reduces Inflammation and Oxidative Stress and Improves Insulin Sensitivity in a Model of Metabolic Syndrome

Chronic, low-level inflammation may be an independent marker of Metabolic Syndrome (MetS). Systemic Enzyme Therapy (SET), the oral administration of proteolytic enzymes, is safe and effective in the management of inflammation. Therefore, the effects of SET, as Wobenzym&reg, on the prevention and treatment of inflammation and other metabolic risk factors were assessed in a rabbit model of diet-induced MetS. Animals were fed a lipid-enriched diet for 8 weeks during which they were administered a vehicle control (control group) or Wobenzym either throughout the study period (prevention group) or beginning at the 5th week, after the development of biomarkers of MetS (treatment group). At the 8th week, both prevention and treatment groups demonstrated improved insulin sensitivity relative to the control group and reduced serum C-reactive protein (CRP) and glycosylated hemoglobin (HbA1c, P < 0.001). At 8 weeks, the prevention group, but not the treatment group, exhibited reduced total cholesterol and oxidative stress, measured as serum malondialdehyde (P < 0.001). Triglycerides and free fatty acids were reduced in both the treatment (P < 0.01) and prevention groups (P < 0.001) relative to the control group at the 8th week. Body weight and blood glucose were not affected. Enzyme therapy may have a positive effect on inflammation, insulin sensitivity, and other metabolic risk factors of MetS.

Effects of Sodium Nitroprussiate on the Metabolites and Key Enzyme Activities of Carbon-nitrogen Metabolism from Seed Germination to Seedling Period of Cherry Tomato

This study was conducted to elucidate the regulating mechanism of sodium nitroprussiate on seed germination and seedling growth of cherry tomato. After the treatment with different concentrations of sodium nitroprusside, the effects on the metabolites and key enzyme activities in carbon-nitrogen metabolism of cherry tomato cuhivar Zhuyun as an experimental material were investigated. The results showed that from seed germination to seedling period, the contents of starch and total nitrogen decreased, but the contents of soluble sugar, reducing sugar, sucrose and soluble protein increased firstly and then decreased; and free amino acids content increased gradually. Sodium nitroprusside made the contents of starch, sucrose and free amino acids higher than CK. From seed germination to seedling period in cherry tomato, the activities of amylase, glutamine synthetase （GS） and sucrose phosphate synthase （SPS） decreased; the activities of nitrate reductase （NR） and glutamate synthase （GOGAT） increased at first and decreased then; and the CK and the 0.25 mmol/L sodium nitroprusside treatments exhibited de- creased sucrose synthase （SS） activity, and the trend was increasing at first and decreasing then after the treatment with 0.50 and 1.00 mmol/L sodium nitroprus- side. Sodium nitroprusside treatment improved amylase activity; and the carbon-nitrogen ratio of the CK increased at first and decreased then, while the values of other treatments tended to decrease. In addition, sodium nitroprusside did not affect seed germination potential and germination rate, but significantly improved biomass accumulation, root length and height of seedlings. These data suggest that sodium nitroprusside could affect the conversion of starch and sugar accumulation, delay the decomposition of total nitrogen and soluble protein, and achieve the effects of accelerating the accumulation of free amino acids, and promoting seed germination and seedling growth, and 0.50 mmol/L sodium nitroprusside has the best effect.

Enzyme Kinetic Equations of Irreversible and Reversible Reactions in Metabolism

This paper compares the irreversible and reversible rate equations from several uni-uni kinetic mechanisms (Michaelis-Menten, Hill and Adair equations) and bi-bi mechanisms (single- and double- displacement equations). In reversible reactions, Haldane relationship is considered to be identical for all mechanisms considered and reversible equations can be also obtained from this rela- tionship. Some reversible reactions of the metabolism are also presented, with their equilibrium constant.