Metabolic Enzyme Activities Research Articles

Feeding frequency does not interact with BPA exposure to influence metabolism or behaviour in zebrafish (Danio rerio)

Resource limitation can constrain energy (ATP) production, and thereby affect locomotion and behaviour such as exploration of novel environments and boldness. Consequently, ecological processes such as dispersal and interactions within and between species may be influenced by food availability. Energy metabolism, and behaviour are regulated by endocrine signalling, and may therefore be impacted by endocrine disrupting compounds (EDCs) including bisphenol A (BPA) derived from plastic manufacture and pollution. It is important to determine the impacts of these novel environmental contexts to understand how human activity alters individual physiology and behaviour and thereby populations. Our aim was to determine whether BPA exposure interacts with feeding frequency to alter metabolism and behaviour. In a fully factorial experiment, we show that low feeding frequency reduced zebrafish (Danio rerio) mass, condition, resting metabolic rates, total distance moved and speed in a novel arena, as well as anxiety indicated by the number of times fish returned to a dark shelter. However, feeding frequency did not significantly affect maximal metabolic rates, aerobic scope, swimming performance, latency to leave a shelter, or metabolic enzyme activities (citrate synthase and lactate dehydrogenase). Natural or anthropogenic fluctuation in food resources can therefore impact energetics and movement of animals with repercussions for ecological processes such as dispersal. BPA exposure reduced LDH activity and body mass, but did not interact with feeding frequency. Hence, behaviour of adult fish is relatively insensitive to disruption by BPA. However, alteration of LDH activity by BPA could disrupt lactate metabolism and signalling and together with reduction in body mass could affect size-dependent reproductive output. BPA released by plastic manufacture and pollution can thereby impact conservation and management of natural resources.

Risk of resistance and the metabolic resistance mechanism of Laodelphax striatellus (Fallén) to cyantraniliprole

Cyantraniliprole is a highly effective diamide insecticide used to control of Laodelphax striatellus (Fallén). This study aimed to assess the insecticide resistance risk of L. striatellus and its metabolic resistance mechanisms. After 25 continuous generations of selection, the resistance of L. striatellus to cyantraniliprole increased by 17.14-fold. The realistic heritability of resistance was 0.0751. After successive rearing for five generations without exposure to insecticides, the resistance ratio for the resistant strain of L. striatellus decreased by 3.47-fold, and the average resistance decline rate per generation was 0.0266. Cyantraniliprole-resistant strains did not exhibit cross-resistance to triflumezopyrim, pymetrozine, flonicamid, sulfoxaflor, dinotefuran, clothianidin, thiamethoxam, nitenpyram, or imidacloprid. Compared to those of the sensitive strain, the 2nd, 3rd, and 4th instars, nymphal stage durations, total preoviposition period, and average generation time of the resistant strain were markedly reduced. Furthermore, the activity of cytochrome P450 monooxygenase (P450) and carboxylesterase (CarE) were markedly increased. The upregulation of CYP419A1v2 expression was most evident among the P450 genes, with a 6.10-fold increase relative to that in the sensitive strain. The CarE gene LsCarE5 was significantly upregulated by 1.94-fold compared with that in the sensitive strain. With the continuous use of cyantraniliprole, L. striatellus may develop resistance to this insecticide. This resistance may be related to the increase in metabolic enzyme activities regulated by the overexpression of P450 and CarE genes.

Associations of phase angle with platelet-activating factor metabolism and related dietary factors in healthy volunteers.

Phase angle (PA) is derived from bioelectrical impedance analysis (BIA). It reflects cell membrane function and decreases in disease. It is affected by inflammation, oxidative stress, and diet. Platelet-activating factor (PAF) is a potent inflammatory lipid mediator. Its levels, along with the activity of its metabolic enzymes, including CDP-choline:1-alkyl-2-acetyl-sn-glycerol-cholinephosphotransferase, acetyl-CoA:lyso-PAF-acetyltransferases, and PAF-AH/Lp-PLA2 are also related to dietary factors, such as the dietary antioxidant capacity (DAC). The aim of the study was to estimate whether the PAF metabolic circuit and related dietary factors are associated with PA in healthy volunteers. In healthy subjects, PAF, its metabolic enzyme activity, and erythrocyte fatty acids were measured, while desaturases were estimated. Food-frequency questionnaires and recalls were used, and food groups, macronutrient intake, MedDietScore, and DAC were assessed. Lifestyle and biochemical variables were collected. DXA and BIA measurements were performed. Lp-PLA2 activity was positively associated with PA (rho = 0.651, p < 0.001, total population; rho = 0.780, p < 0.001, women), while PAF levels were negatively associated with PA only in men (partial rho = -0.627, p = 0.012) and inversely related to DAC. Estimated desaturase 6 was inversely associated with PA (rho = -0.404, p = 0.01, total sample). Moreover, the DAC correlated positively with PA (rho = 0.513, p = 0.03, women). All correlations were adjusted for age, body mass index, and sex (if applicable). PA is associated with PAF levels and Lp-PLA2 activity in a gender-dependent fashion, indicating the involvement of PAF in cell membrane impairment. The relationship of PA with DAC suggests a protective effect of antioxidants on cellular health, considering that antioxidants may inhibit PAF generation.

Imaging the skin epilipidome and the activity of metabolic key enzymes in the senescence process at single cell level

Imaging the skin epilipidome and the activity of metabolic key enzymes in the senescence process at single cell level

Biochemical and Ultrastructural Changes in Wheat Plants during Drought Stress.

Drought severely slows down plant growth, decreases crop yield, and affects various physiological processes in plants. We examined four local bread wheat cultivars with different drought tolerance (drought-tolerant Zirva 85 and Murov 2 and drought-sensitive Aran and Gyzyl bughda cultivars). Leaves from seedlings of drought-tolerant plants demonstrated higher activity of antioxidant enzymes and lower levels of malondialdehyde and hydrogen peroxide. The content of soluble proteins in drought-exposed increased, possibly due to the stress-induced activation of gene expression and protein synthesis. Drought-exposed Zirva 85 plants exhibited an elevated activity of nitrogen and carbon metabolism enzymes. Ultrastructural analysis by transmission electron microscopy showed drought-induced damage to mesophyll cells and chloroplast membranes, although it was manifested less in the drought-tolerant cultivars. Comparative analysis of the activity of metabolic and antioxidant enzymes, as well as observed ultrastructural changes in drought-exposed plants revealed that the response to drought of seedlings was more pronounced in drought-tolerant cultivars. These findings can be used in further studies of drought stress in wheat plants under natural conditions.

Effects of LED spectra on the growth and physiological mechanism of juvenile Sebastes schlegelii. Part I: Growth, feeding and digestion and metabolism

Light is a crucial and multifaceted ecological factor that profoundly influences fish feeding, growth, development, and physiological processes, either directly or indirectly. To examine how various LED spectra affect the growth, feeding, and digestion of juvenile Sebastes schlegelii, we conducted this study using juvenile Sebastes schlegelii specimens with an initial body length of (6.27 ± 0.05) cm and an initial body weight of (7.56 ± 0.17) g. In this experiment, 750 juvenile Sebastes schlegelii were carefully selected and placed into five distinct LED spectral treatment groups: green light (GL, λ 525–530 nm), blue light (BL, λ 450–455 nm), yellow light (YL, λ 590–595 nm), red light (RL, λ 625–630 nm), and white light (WL, λ 400–780 nm). These fish were closely monitored throughout the study. The photoperiod was set at 12 h of light followed by 12 h of darkness (12 L:12 D), and the light intensity was maintained at 250 mW/m2. After a 60-day experimental period, we evaluated the growth performance, feeding intake, as well as the activities of digestive and metabolic enzymes. The findings indicated that juvenile Sebastes schlegelii exhibited improved length, weight gain, fatness, and feed conversion efficiency under the illumination conditions of GL, BL, and WL. At the same time, RL had a detrimental impact on the growth, development, and feeding performance of juvenile Sebastes schlegelii, resulting in reduced body length and weight, as well as the lowest fattening and feed conversion efficiency. Regarding digestive enzyme activity, juvenile Sebastes schlegelii reared under BL and YL conditions exhibited higher digestive enzyme activity. The GL, BL, and YL environments significantly increased the metabolic enzyme activity of juvenile Sebastes schlegelii, which reached a higher level by the end of the experiment. The results indicate that, when compared to other spectral environments, GL, BL, and WL were advantageous for the growth, feeding, hunting, digestion, and metabolism of juvenile Sebastes schlegelii. These conditions promoted healthy growth and a more favorable growth trend in juvenile fish. In contrast, RL hindered the growth of juvenile fish.

Differential effects of Western diet and traumatic muscle injury on skeletal muscle metabolic regulation in male and female mice.

This study was designed to develop an understanding of the pathophysiology of traumatic muscle injury in the context of Western diet (WD; high fat and high sugar) and obesity. The objective was to interrogate the combination of WD and injury on skeletal muscle mass and contractile and metabolic function. Male and female C57BL/6J mice were randomized into four groups based on a two-factor study design: (1) injury (uninjured vs. volumetric muscle loss [VML]) and (2) diet (WD vs. normal chow [NC]). Electrophysiology was used to test muscle strength and metabolic function in cohorts of uninjured+NC, uninjured+WD, VML+NC and VML+WD at 8weeks of intervention. VML-injured male and female mice both exhibited decrements in muscle mass (-17%, P<0.001) and muscle strength (-28%, P<0.001); however, VML+WD females had a 28% greater muscle mass compared to VML+NC females (P=0.034), a compensatory response not detected in males. VML-injured male and female mice both had lower carbohydrate- and fat-supported muscle mitochondrial respiration (JO2 ) and less electron conductance through the electron transport system (ETS); however, male VML-WD had 48% lower carbohydrate-supported JO2 (P=0.014) and 47% less carbohydrate-supported electron conductance (P=0.026) compared to male VML+NC, and this diet-injury phenotype was not present in females. ETS electron conductance starts with complex I and complex II dehydrogenase enzymes at the inner mitochondrial membrane, and male VML+WD had 31% less complex I activity (P=0.004) and 43% less complex II activity (P=0.005) compared to male VML+NC. This was a diet-injury phenotype not present in females. Pyruvate dehydrogenase (PDH), β-hydroxyacyl-CoA dehydrogenase, citrate synthase, α-ketoglutarate dehydrogenase and malate dehydrogenase metabolic enzyme activities were evaluated as potential drivers of impaired JO2 in the context of diet and injury. There were notable male and female differential effects in the enzyme activity and post-translational regulation of PDH. PDH enzyme activity was 24% less in VML-injured males, independent of diet (P<0.001), but PDH enzyme activity was not influenced by injury in females. PDH enzyme activity is inhibited by phosphorylation at serine-293 by PDH kinase 4 (PDK4). In males, there was greater total PDH, phospho-PDHser293 and phospho-PDH-to-total PDH ratio in WD mice compared to NC, independent of injury (P≤0.041). In females, PDK4 was 51% greater in WD compared to NC, independent of injury (P=0.025), and was complemented by greater phospho-PDHser293 (P=0.001). Males are more susceptible to muscle metabolic dysfunction in the context of combined WD and traumatic injury compared to females, and this may be due to impaired metabolic enzyme functions.

Effect of alpha-ketoglutarate supplementation on growth, antioxidant capacity, gene expression and amino acid profile in Nile tilapia (Oreochromis niloticus) under varying protein diets

Protein is the most expensive component in aquafeed and emphasis is given to reduce dietary protein levels without compromising growth. Alpha ketoglutarate (AKG) promotes protein synthesis in fish and improves growth. The present experiment was conducted to investigate the effect of AKG with different dietary protein levels on growth, nutrient utilisation, antioxidant status and immunity in Nile tilapia (Oreochromis niloticus). Five diets were prepared with different levels of protein such as 20 (P20A), 25 (P25A), 30 (P30A) and 35 % (P35A) with 1 % AKG and 35 % protein without AKG (P35C). Nile tilapia fingerlings (300) were randomly stocked in fifteen 150 L capacity FRP tanks with 20 fingerlings (9.91 ± 0.06 g) in each. The fish were then fed with experimental diets twice daily for 60 days. The fish fed with P30A had significantly (P < 0.05) higher growth performance and nutrient utilisation. However, there was no significant (P > 0.05) difference in final body weight and weight gain of tilapia fed with P35C and P25Adiets. The feed conversion ratio (FCR) was significantly higher (P < 0.05) in P20A group and other groups showed alike (P > 0.05) FCR value. The whole body protein and lipid contents of final fish were significantly (P < 0.05) higher in P30A and P25A, respectively. The P25A, P30A and P35A showed increased (P < 0.05) activities of superoxide dismutase (SOD) and catalase (CAT). The activity of metabolic enzymes were also higher in higher protein levels with AKG (P30A and P35A). The expression level of growth gene insulin like growth factor 1 (IGF-1) was significantly up-regulated (P < 0.05) in the P30A. In addition, the relative expression levels of the pro-inflammatory factors tumour necrosis factor (TNF-α) were significantly (P < 0.05) lower in P30A, P25A, and P20A. Total amino acid profile of fish was higher in P35C group. In summary, the optimal choice for enhancing growth and health in Nile tilapia appears to be a diet comprising 30 % protein and 1 % AKG. However, diet of 25 % CP with AKG showed equivalent result in terms growth that of 35 % CP level without AKG. Hence, 25 % protein with 1 % AKG can effectively fulfil protein requirement in tilapia.

Nanozyme-Based Regulation of Cellular Metabolism and Their Applications.

Metabolism is the sum of the enzyme-dependent chemical reactions, which produces energy in catabolic process and synthesizes biomass in anabolic process, exhibiting high similarity in mammalian cell, microbial cell, and plant cell. Consequently, the loss or gain of metabolic enzyme activity greatly affects cellular metabolism. Nanozymes, as emerging enzyme mimics with diverse functions and adjustable catalytic activities, have shown attractive potential for metabolic regulation. Although the basic metabolic tasks are highly similar for the cells from different species, the concrete metabolic pathway varies with the intracellular structure of different species. Here, the basic metabolism in living organisms is described and the similarities and differences in the metabolic pathways among mammalian, microbial, and plant cells and the regulation mechanism are discussed. The recent progress on regulation of cellular metabolism mainly including nutrient uptake and utilization, energy production, and the accompanied redox reactions by different kinds of oxidoreductases and their applications in the field of disease therapy, antimicrobial therapy, and sustainable agriculture is systematically reviewed. Furthermore, the prospects and challenges of nanozymes in regulating cell metabolism are also discussed, which broaden their application scenarios.

New palladium complexes with N-heterocyclic carbene and morpholine ligands: Synthesis, characterization, crystal structure, molecular docking, and biological activities.

This work includes the synthesis of a new series of palladium-based complexes containing both morpholine and N-heterocyclic carbene (NHC) ligands. The new complexes were characterized using NMR (1 H and 13 C), FTIR spectroscopic, and elemental analysis techniques. The crystal structure of complex 1b was obtained by utilizing the single-crystal X-ray diffraction method. X-ray studies show that the coordination environment of palladium atom is completed by the carbene carbon atom of the NHC ligand, the nitrogen atom of the morpholine ring, and a pair of bromide ligand, resulting in the formation of slightly distorted square planar geometry. All complexes were determined for some metabolic enzyme activities. Results indicated that all the synthetic complexes exhibited powerful inhibitory actions against all aims as compared to the control molecules. Ki values of new morpholine-liganded complexes bearing 4-hydroxyphenylethyl group 1a-e for hCA I, hCA II, AChE, BChE, and α-glycosidase enzymes were obtained in the ranges 0.93-2.14, 1.01-2.03, 4.58-10.27, 7.02-13.75, and 73.86-102.65 µM, respectively. Designing of reported complexes is impacted by molecular docking study, and interaction with the current enzymes also proclaimed that compounds 1e (-12.25 kcal/mol for AChE and -11.63 kcal/mol for BChE), 1c (-10.77 kcal/mol and -9.26 kcal/mol for α-Gly and hCA II, respectively), and 1a (-8.31 kcal/mol for hCA I) are showing binding affinity and interaction from the synthesized five novel complexes.

Using regular and transcriptomic analyses to investigate the biotransformation mechanism and phytotoxic effects of 6:2 fluorotelomer carboxylic acid (6:2 FTCA) in pumpkin (Cucurbita maxima L.)

Although 6:2 fluorotelomer carboxylic acid (6:2 FTCA), which is one of the most popular substitutes for perfluorooctanoic acid (PFOA), has been widely distributed in environments, little is known about its biotransformation mechanism and phytotoxic effects in plants. Here, we showed that 6:2 FTCA could be taken up by pumpkin (Cucurbita maxima L.) roots from exposure solution and acropetally translocated to shoots. Biotransformation of 6:2 FTCA to different carbon chain perfluorocarboxylic acid (PFCA) metabolites (C2-C7) via α-and β-oxidation in pumpkin was observed, and perfluorohexanoic acid (PFHxA) was the major transformation product. The results of enzyme assays, enzyme inhibition experiments and gene expression analysis indicated that cytochrome P450 (CYP450), glutathione-S-transferase (GST) and ATP-binding cassette (ABC) transporters were involved in the metabolism of 6:2 FTCA in pumpkin. Plant-associated rhizobacteria and endophyte also contributed to 6:2 FTCA degradation through β-oxidation. The chlorophyll (Chl) content and genes involved in photosynthesis were significantly improved by 6:2 FTCA. The reductions of antioxidant and metabolic enzyme activities reflected the antioxidant defense system and detoxification system of pumpkin were both damaged, which were further confirmed by the down-regulating associated genes encoding phenylpropanoid biosynthesis, endoplasmic reticulum-related proteins, ascorbate-glutathione cycle and ABC transporters. This study is helpful to understand the environmental behaviors and toxicological molecular mechanisms of 6:2 FTCA in plants.

Taurine promotes muscle fiber type transformation through CaN/NFATc1 signaling in porcine myoblasts.

This study investigated the effect of taurine (TAU) on the muscle fiber type transformation in porcine myoblasts and its molecular mechanisms. The findings revealed that TAU augmented the protein expression of slow MyHC and the enzyme activities of oxidative metabolism markers like malate dehydrogenase and succinic dehydrogenase. Conversely, it curtailed the expression of fast MyHC and glycolytic metabolism enzyme activity of lactate dehydrogenase. Moreover, TAU elevated the expression of genes associated with oxidative fiber while diminishing the expression of those linked to glycolytic fibers, suggesting that TAU promoted the muscle fiber type transformation from glycolytic fiber to oxidative fiber. Additionally, TAU notably enhanced the expression of key molecules of calcineurin (CaN)/nuclear factor of activated T cells c1 (NFATc1) signaling and the CaN activity in porcine myoblasts. However, CaN inhibitor cyclosporine A abolished these effects induced by TAU. Our results indicated that TAU regulated the muscle fiber type transformation from glycolytic to oxidative fiber by activation of CaN/NFATc1 signaling.

Opioid infusions at different times of the day produce varying degrees of opioid-induced hyperalgesia

BackgroundOpioids are metabolised by enzymes the activities of which vary with the circadian rhythm. We examined whether opioid infusions administered at different times of the day produce varying degrees of opioid-induced hyperalgesia (OIH) in animal experiments and clinical studies. MethodsMale Sprague–Dawley rats received remifentanil infusions (1 μg kg−1·min−1 for 1 h) at Zeitgeber times (ZT) 0, 4, 8, 12, 16, or 20 h. Rhythmicity of mechanical hypersensitivity was assayed after the infusion. Mechanical hypersensitivity, drug concentration, and metabolic enzyme activity of Wistar rats that received sufentanil (10 μg kg−1; four consecutive i.p. injections at 15-min intervals) or remifentanil infusion at ZT0 or ZT8 were assayed. Sixty patients who underwent abdominal laparoscopic surgery under general anaesthesia received remifentanil infusion (0.15 μg kg−1 min−1) and sufentanil injection (0.2 μg kg−1) at induction and skin incision, respectively. Postoperative pressure pain sensitivity, pain Numeric Rating Scale (NRS), drug concentrations, and nonspecific esterase activity were assessed. ResultsSprague–Dawley rats that received remifentanil infusion exhibited a robust rhythmic paw withdrawal threshold (JTK_CYCLE: P=0.001, Q=0.001, Phase=26). Wistar rats infused with remifentanil or sufentanil at ZT8 exhibited greater OIH (P<0.001) than those infused at ZT0, with higher blood concentrations (P<0.001) and lower metabolic enzyme activities (P=0.026 and P=0.028, respectively). Patients in the afternoon group exhibited higher pressure pain sensitivity at forearm (P=0.002), higher NRS (P<0.05), higher drug concentrations (sufentanil: P=0.037, remifentanil: P=0.005), and lower nonspecific esterase activity (P=0.024) than the morning group. ConclusionsOpioid infusions administered at different times of day produced varying degrees of OIH, possibly related to circadian rhythms of metabolic enzyme activities. Clinical trial registrationNCT 05234697.

Comparative Study on the Resistance of Beta-Cypermethrin Nanoemulsion and Conventional Emulsion in Blattella germanica.

This study aimed to compare the resistance rates of Blattella germanica to beta-cypermethrin nanoemulsion and conventional emulsion and establish reference values via biochemical detection for conventional emulsion. We conducted experiments using subcultured Blattella germanica and applied the micro-drop method for treatment. Subsequently, the activity of metabolic enzymes was measured using spectrophotometry. Profile analysis was employed to study the resistance rates of beta-cypermethrin nanoemulsion and beta-cypermethrin emulsion. The regression equation for the relationship between generation and resistance factor in Blattella germanica treated with beta-cypermethrin nanoemulsion was as follows: y1 = 0.091x1 + 0.991, with an r-value of 0.990 (F = 95.184, p = 0.01 < 0.05). Similarly, the regression equation for Blattella germanica treated with emulsion was y2 = 0.376x2 + 1.051, with an r-value of 0.993 (F = 141.094, p = 0.007 < 0.05). The comparison of slopes between these two regression equations yielded an F-value of 8.61, indicating a significant difference (p = 0.001 < 0.05). Our findings suggest that the resistance factor in Blattella germanica treated with beta-cypermethrin nanoemulsion differs from that treated with beta-cypermethrin emulsion. Specifically, the resistance factor of beta-cypermethrin nanoemulsion increased at a slower rate compared to beta-cypermethrin emulsion.

The Potential Hepatoprotective Effect of Vaccinium arctostaphylos L. Fruit Extract in Diabetic Rat.

Vaccinium arctostaphylos has traditionally been employed in Iranian folk medicine to treat diabetes. However, the precise molecular mechanisms underlying its antidiabetic properties remain incompletely understood. The current experiment intended to explore the modulatory effects of V. arctostaphylos fruit ethanolic extract (VAE) on biochemical and molecular events in the livers of diabetic rats. In this experimental study, male Wistar rats were randomly assigned to four groups: normal control, normal rats with VAE treatment, diabetic control, and diabetic rats with VAE treatment. Following 42 days of treatment, the impact of VAE on diabetes-induced rats was assessed by measuring various serum biochemical parameters, including insulin, free fatty acids (FFA), tumor necrosis factor-α (TNF-α), reactive oxygen species (ROS), and adiponectin levels. The activities of hepatic carbohydrate metabolic enzymes and glycogen content were determined. Additionally, expression levels of selected genes implicated in carbohydrate/lipid metabolism and miR-27b expression were evaluated. H and E-stained liver sections were prepared for light microscopy examination. Treatment with VAE elevated levels of insulin and adiponectin that reduced levels of FFA, ROS, and TNF-α in the serum of diabetic rats. VAE-treated rats exhibited increased activities of hepatic glucokinase (GK), glucose-6-phosphate dehydrogenase (G6PD), and glycogen concentrations, in conjunction with decreased activities of glucose-6-phosphatase (G6Pase) and fructose-1,6-bisphosphatase (FBPase). Furthermore, VAE significantly upregulated the transcription levels of hepatic insulin receptor substrate 1 (Irs1) and glucose transporter 2 (Glut2), while considerably downregulated the expression of peroxisome proliferator-activated receptor gamma (Pparg) and sterol regulatory element-binding protein 1c (Srebp1c). VAE remarkably enhanced the expression of miR27-b in the hepatic tissues of diabetic rats. Abnormal histological signs were dramatically normalized in diabetic rats receiving VAE compared to those in the diabetic control group. Our findings underscore the hypoglycemic and hypolipidemic activities of V. arctostaphylos and assist in better comprehension of its antidiabetic properties.

Effect of MTA3 Inhibition of Glutamine Synthetase-Mediated Glutaminolysis on Radiosensitivity of Patients with Esophageal Cancer

Metastasis-associated protein 3 (MTA3) can serve as a tumor suppressor in many cancer types. However, the role of MTA3 in radiosensitivity of patients with esophageal squamous cell cancer (ESCC) remains unclear. We thus investigated the function of MTA3 in radiosensitivity for ESCC, one of the most common digestive cancers. The colony formation assay and nude mice xenograft tumor assay were performed to investigate the effect of MTA3 on radiosensitivity in ESCC. Glutamine consumption assay kit and glutamate production assay kit were used to assess the glutaminolysis. Glutaminase (GLS) Activity Assay Kit and Glutamine Synthetase (GS) Activity Assay Kit were used to analyze the activity of specific metabolic enzymes dominate glutaminolysis. The regulatory mechanism of glutaminolysis by MTA3 was confirmed using Chromatin immunoprecipitation assay and Gaussia luciferase assay. The expression levels of MTA3 and GS in ESCC primary tissues were evaluated using immunohistochemistry. Survival curves were plotted with the Kaplan-Meier method and compared by log-rank test. The colony formation assay showed that MTA3 depletion and overexpression caused significantly higher and lower clonogenic survival after different doses of irradiation (IR), respectively. When these cells were subcutaneously injected into nude mice, the tumors derived from the cells with MTA3 overexpression and MTA3 knockdown were significantly smaller and bigger after IR, respectively. These findings suggest that MTA3 can enhance radiosensitivity in vitro and in vivo. Meanwhile, overexpressed and knockdown MTA3 can repress and expedite glutamine consumption and glutamate production uniformly, respectively. To determine how MTA3 acts on glutaminolysis, the activity of two specific metabolic enzymes dominate this metabolism, GS and GLS, were evaluated. It found that overexpressed and knockdown MTA3 can restrain and enhance the activity of GS, respectively, but have less effect on GLS. Moreover, the decreased radiosensitivity mediated by MTA3 knockdown is significantly increased when treated with GS inhibitor, suggesting that GS plays a crucial role in MTA3-mediated radiosensitivity enhancement. Mechanistically, Chromatin immunoprecipitation assay and Gaussia luciferase assay showed that MTA3 was recruited to the promoter of GS and suppressed GS transcription. However, knockdown of GATA3 abolished MTA3's repressive effect on GS and inhibited the MTA3's occupation on the promoter region of GS. These results collectively demonstrated that, in ESCC cells, MTA3 is recruited by GATA3 to inhibit GS expression, then ultimately represses glutaminolysis and enhances radiosensitivity. Finally, we showed that the ESCC patients in the MTA3low/GShigh group is significantly associated with shorter overall survival. MTA3 is capable of enhancing radiosensitivity through downregulating GS and MTA3low/GShigh might be a potential prognostic factor for ESCC patients.

Mechanical stiffness promotes skin fibrosis through Piezo1-mediated arginine and proline metabolism

The increased mechanics of fibrotic skin tissue continuously regulate fibroblast functions such as survival and differentiation. Although all these processes consume metabolites, it is unclear whether and how cells adapt their metabolic activity to increased matrix stiffness. Here, we show that transferring mouse dermal fibroblasts from soft to stiff substrates causes an up-regulation of arginine and proline metabolism. Increased matrix stiffness stimulates the expression and activity of key metabolic enzymes, leading to the synthesis of L-proline, a major source of collagen. In addition, the novel mechanosensitive channel Piezo1 was identified as a key regulator of arginine and proline metabolism in fibroblasts under increased stiffness. Consistently, targeting Piezo1 to dermal fibroblasts in vivo effectively reduces fibrosis and arginine-proline metabolism in mouse skin. Therefore, mechanical stiffness is a critical environmental cue for fibroblast metabolism and skin fibrosis progression.

Protective effect of bromelain on some metabolic enzyme activities in tyloxapol-induced hyperlipidemic rats.

Elevation of one or more plasma lipids, such as phospholipids, cholesterol esters, cholesterol, and triglycerides, is known as hyperlipidemia. In humans and experimental animals, bromelain, the primary active ingredient isolated from pineapple stems, has several positive effects, including anti-tumor growth, anticoagulation, and anti-inflammation. Hence, the purpose of this study was to determine the possible protective impact of bromelain on some metabolic enzymes (paraoxonase-1, glutathione S-transferase, glutathione reductase, sorbitol dehydrogenase [SDH], aldose reductase [AR], butyrylcholinesterase [BChE], and acetylcholinesterase [AChE]), activity in the heart, kidney, and liver of rats with tyloxapol-induced hyperlipidemia. Rats were divided into three groups: control group, HL-control group (tyloxapol 400mg/kg, i.p. administered group), and HL+bromelain (group receiving bromelain 250mg/kg/o.d. prior to administration of tyloxapol 400mg/kg, i.p.). BChE, SDH, and AR enzyme activities were significantly increased in all tissues in HL-control compared to the control, whereas the activity of other studied enzymes was significantly decreased. Bromelain had a regulatory effect on all tissues and enzyme activities. In conclusion, these results prove that bromelain is a new mediator that decreases hyperlipidemia.

Synthesis, characterization, biochemical, and molecular modeling studies of carvacrol-based new thiosemicarbazide and 1,3,4-thiadiazole derivatives.

A series of carvacrol-based thiosemicarbazide (3a-e) and 1,3,4-thiadiazole-2-amine (4a-e) were designed and synthesized for the first time. The structures were characterized by nuclear magnetic resonanceand high resolution mass spectroscopytechniques. All compounds were examined for some metabolic enzyme activities. Results indicated that all the synthetic molecules exhibited powerful inhibitory actions against human carbonic anhydrase I and II (hCAI and II), acetylcholinesterase (AChE), and butyrylcholinesterase (BChE) enzymes compared to the standard molecules. Ki values of five novel thiosemicarbazides and five new 1,3,4-thiadiazole-2-amine derivatives (3a-e and 4a-e) for hCA I, hCA II, AChE, and BChE enzymes were obtained in the ranges 0.73-21.60, 0.42-15.08 µM, 3.48-81.48, 92.61-211.40 nM, respectively. After the experimental undertaking, an extensive molecular docking analysis was conducted to scrutinize the intricate details of interactions between the ligand and the enzyme in question. The principal focus of this investigation was to appraise the potency and efficacy of the most active compound. In this context, the calculated docking scores were noted to be remarkably low, with values of -8.65, -7.97, -8.92, and -8.32 kcal/mol being recorded for hCA I, hCA II, AChE, and BChE, respectively. These observations suggest a high affinity and specificity of the studied compounds towardthe enzymes, as mentioned earlier, which may pave the way for novel therapeutic interventions aimed at modulating the activity of these enzymes.

Effects of Arbutin on Potassium Bromate-Induced Erythrocyte Toxicity in Rats: Biochemical Evaluation of Some Metabolic Enzyme Activities In Vivo and In Vitro.

In the present study, the inhibitory effect of potassium bromate on the pentose phosphate pathway and intracellular antioxidant systems enzymes (glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), glutathione reductase (GR), glutathione S-transferase (GST), and thioredoxin reductase (TrxR)) and the role of arbutin in ameliorating this inhibition were investigated. In the in vivo phase of the study, Wistar Albino rats (28 male adults) were randomly divided into four groups. Control (n = 7): isotonic serum (0.5 mL, i.p), potassium bromate group (n = 7): potassium bromate (100 mg/kg), arbutin group (n = 7): arbutin (i.p.) (50 mg/kg/day), potassium bromate + arbutin, and Group (n = 7): potassium bromate (100 mg/kg) + arbutin (50 mg/kg/day) (i.p). The results of in vivo study showed that the activities of G6PD, 6PGD, GR, and TrxR enzymes were strongly inhibited in potassium bromate groups (p < 0.05). It was determined that GST enzyme activity decreased in the potassium bromate group, but this decrease was not statistically significant compared to the control group (p ⩾ 0.05). A statistically significant increase was found in G6PD, 6PGD, GST, and TrxR enzyme activities in the arbutin group compared to the control group (p < 0.05). The increase in GR enzyme activity was not statistically significant (p ⩾ 0.05). The potassium bromate + arbutin group's enzyme activity increased in comparison to the potassium bromate group and was discovered to be closer to the control group. It was found that potassium bromate inhibited the 6PGD enzyme obtained from rat erythrocyte tissues with IC50 = 346 μM value and Ki = 434.4 μM ± 6.1 value, and the inhibition was noncompetitive.