Expression Of Related Enzymes Research Articles

Storage Temperature Affects the Accumulation of Phenolics in Fresh-Cut Melon by Accelerating the Sucrose Decomposition

To accurately evaluate the role of storage temperature in improving the quality of fresh-cut fruits and vegetables, the effects of two storage temperatures (5 °C and 15 °C) on the phenylpropanoid pathway and sucrose metabolism in fresh-cut melon (cv. Yugu) cubes were determined. A higher temperature (15 °C) expedited sucrose decomposition in the melon cubes at the early stage of storage, resulting in higher levels of glucose and fructose. This effect was corroborated by increased activities of acid invertase (AI), neutral invertase (NI), and sucrose synthase cleavage (SS-c), along with higher expressions of CmAI1/2, CmNI1/2, and CmSS1/2 in the melon cubes at 15 °C. Additionally, the higher activity and gene expression of hexokinase in melon cubes at 15 °C led to an increase in the utilization rate of sugars toward downstream metabolic pathways. Moreover, the melon cube storage at 15 °C elevated the activities and gene expressions of phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), and 4-coumaric acid: CoA ligase (4CL), thereby increasing the synthesis of phenolics. Sucrose showed a significant negative correlation with PAL, C4H, and 4CL, as well as with CmPAL5/7 and CmC4H1/3. However, hexokinase displayed a significant positive correlation with PAL, C4H, and 4CL, as well as with CmPAL1, CmPAL3-9, CmC4Hs, and Cm4CLs. These findings demonstrate that a higher-temperature storage of melon cubes can accelerate the phenylpropanoid pathway and sucrose metabolism by regulating the activity and gene expression of related enzymes, thereby inducing phenolic accumulation. These results also indicate that lower-temperature storage is not conducive to the conversion of sugars into phenolics in fresh-cut melon. Therefore, the temperature can be appropriately and briefly raised in the production and preservation process of fresh-cut melon to obtain higher levels of phenolics.

P53 activation enhances the sensitivity of non-small cell lung cancer to the combination of SH003 and docetaxel by inhibiting de novo pyrimidine synthesis

BackgroundIdentifying molecular biomarkers for predicting responses to anti-cancer drugs can enhance treatment precision and minimize side effects. This study investigated the novel cancer-targeting mechanism of combining SH003, an herbal medicine, with docetaxel in non-small cell lung cancer (NSCLC) cells. Also, the present study aimed to identify the genetic characteristics of cancer cells susceptible to this combination.MethodsCell viability was analyzed by WST-8 assay. Apoptosis induction, BrdU incorporation, and cell cycle analysis were performed using flow cytometry. Metabolites were measured by LC–MS/MS analysis. Real-time PCR and western blotting evaluated RNA and protein expression. DNA damage was quantified through immunofluorescence. cBioPortal and GEPIA data were utilized to explore the mutual co-occurrence of TP53 and UMPS and UMPS gene expression in NSCLC.ResultsThe combination treatment suppressed de novo pyrimidine nucleotide biosynthesis by reducing the expression of related enzymes. This blockade of pyrimidine metabolism led to DNA damage and subsequent apoptosis, revealing a novel mechanism for inducing lung cancer cell death with this combination. However, some lung cancer cells exhibited distinct responses to the combination treatment that inhibited pyrimidine metabolism. The differences in sensitivity in lung cancer cells were determined by the TP53 gene status. TP53 wild-type lung cancer cells were effectively inhibited by the combination treatment through p53 activation, while TP53 mutant- or null-type cells exhibited lower sensitivity.ConclusionsThis study, for the first time, established a link between cancer cell genetic features and treatment response to simultaneous SH003 and docetaxel treatment. It highlights the significance of p53 as a predictive factor for susceptibility to this combination treatment. These findings also suggest that p53 status could serve as a crucial criterion in selecting appropriate therapeutic strategies for targeting pyrimidine metabolism in lung cancer.

Impaired cardiac glycolysis and glycogen depletion are linked to poor myocardial outcomes in juvenile male swine with metabolic syndrome and ischemia.

Obesity continues to rise in the juveniles and obese children are more likely to develop metabolic syndrome (MetS) and related cardiovascular disease. Unfortunately, effective prevention and long-term treatment options remain limited. We determined the juvenile cardiac response to MetS in a swine model. Juvenile male swine were fed either an obesogenic diet, to induce MetS, or a lean diet, as a control (LD). Myocardial ischemia was induced with surgically placed ameroid constrictor on the left circumflex artery. Physiological data were recorded and at 22 weeks of age the animals underwent a terminal harvest procedure and myocardial tissue was extracted for total metabolic and proteomic LC/MS-MS, RNA-seq analysis, and data underwent nonnegative matrix factorization for metabolic signatures. Significantly altered in MetS versus. LD were the glycolysis-related metabolites and enzymes. In MetS compared with LD Glycogen synthase 1 (GYS1)-glycogen phosphorylases (PYGM/PYGL) expression disbalance resulted in a loss of myocardial glycogen. Our findings are consistent with the concept that transcriptionally driven myocardial changes in glycogen and glucose metabolism-related enzymes lead to a deficiency of their metabolite products in MetS. This abnormal energy metabolism provides insight into the pathogenesis of the juvenile heart in MetS. This study reveals that MetS and ischemia diminishes ATP availability in the myocardium via altering the glucose-G6P-pyruvate axis at the level of metabolites and gene expression of related enzymes. The observed severe glycogen depletion in MetS coincides with disbalance in expression of GYS1 and both PYGM and PYGL. This altered energy substrate metabolism is a potential target of pharmacological agents for improving juvenile myocardial function in MetS and ischemia.

Low-temperature at booting reduces starch content and yield of wheat by affecting dry matter transportation and starch synthesis.

With the continuous change of global climate, the frequency of low-temperature stress (LTS) in spring increased greatly, which led to the increase of wheat yield decline. The effects of LTS at booting on grain starch synthesis and yield were examined in two wheat varieties with differing low-temperature sensitivities (insensitive variety Yannong 19 and sensitive variety Wanmai 52). A combination of potted and field planting was employed. For LTS treatment at booting, the wheat plants were placed in a climate chamber for 24h at -2°C, 0°C or 2°C from 19:00 to 07:00 then 5°C from 07:00 to 19:00. They were then returned to the experimental field. The effects of flag leaf photosynthetic characteristics, the accumulation and distribution of photosynthetic products, enzyme activity related to starch synthesis and relative expression, the starch content, and grain yield were determined. LTS at booting caused a significant reduction in the net photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr) of the flag leaves at filling. The development of starch grains in the endosperm is also hindere, there are obvious equatorial grooves observed on the surface of the A-type starch granules, and a reduction in the number of B-type starch granules. The abundance of 13C in the flag leaves and grains decreased significantly. LTS also caused a significant reduction in translocation amount of pre-anthesis stored dry matte from vegetative organs to grains and amount of post-anthesis transfer of accumulated dry matte into grains, and the distribution rate of dry matter in the grains at maturity. The grain filling time was shortened, and the grain filling rate decreased. A decrease in the activity and relative expression of enzymes related to starch synthesis was also observed, with a decrease in the total starch content. As a result, a decrease in the grain number per panicle and 1000-grain weight were also observed. These findings highlight the underlying physiological cause of decreased starch content and grain weight after LTS in wheat.

Synthetic Translational Regulation by Protein-Binding RNA Origami Scaffolds.

Rational design approaches for the regulation of gene expression are expanding the synthetic biology toolbox. However, only a few tools for regulating gene expression at the translational level have been developed. Here, we devise an approach for translational regulation using the MS2 and PP7 aptamer and coat-protein pairs in Escherichia coli. The aptamers are used as operators in transcription units that encode proteins fused to their cognate coat proteins, which leads to self-repression. RNA origami scaffolds that contain up to four aptamers serve as an alternate binder to activate translation. With this system, we demonstrate that the increase in expression of a reporter protein is dependent on both the concentration and number of aptamers on RNA origami scaffolds. We also demonstrate regulation of multiple proteins using a single MS2 coat protein fusion and apply this method to regulate the relative expression of enzymes of the branched pathway for deoxyviolacein biosynthesis.

The plant growth-promoting endophytic Fusarium oxysporum GG22 enhances Rehmannia glutinosa secondary metabolites accumulation

Fusarium oxysporum GG22 is an endophytic fungus isolated from the leaves of Rehmannia glutinosa and shown to promote the growth of R. glutinosa. There is currently no in-depth study on the effect of plant growth-promoting endophytic fungi on the defensive metabolic response of medicinal herbs.A symbiotic system of GG22 with R. glutinosa tissue culture seedlings was established to analyze the effect of GG22 infection on the biomass of R. glutinosa tissue culture seedlings and on the accumulation of secondary metabolites. Antioxidant enzymes (SOD, POD and CAT) and endogenous signaling molecules (NO and H2O2) were examined. Quantitative real-time PCR was used to analyze the expression of enzyme genes related to secondarymetabolites. GG22 promoted the growth of the plant height, leaf area, fresh weight, and dry weight of R. glutinosa tissue culture seedlings and significantly increased catalpol and verbascoside. After GG22 infection, the activities of the antioxidant enzyme system (SOD, POD, CAT) of R. glutinosa increased differentially. In addition, significant accumulation of NO and H2O2 signaling molecules occurred. According to the analysis of the expression of genes encoding key biosynthesis enzymes for iridoid glycoside and verbascoside components in R. glutinosa, the levels of AACT, HMGR, DXR, GS, 4CL, C4H and C3H were all significantly increased after GG22 infection. Therefore, it is speculated that infection with the endophytic fungus GG22 induces a burst of reactive oxygen species and NO in plants to activate or enhance the expression of related enzymes in primary metabolic pathways, thereby increasing secondary metabolite contents in R. glutinosa. Our results confirmed that GG22 not only contributes to the stimulation of R. glutinosa growth but also enhances secondary metabolism, thus demonstrating its application potential as a bio-fertilizer for R. glutinosa cultivation.

Bioavailability evaluation of the intestinal absorption and liver accumulation of torularhodin using a rat postprandial model.

Torularhodin, as a new functional carotenoid, possesses great application potential in disease intervention. However, its absorption process and corresponding mechanism have not been studied. In this study, a rat postprandial model was established to explore the absorption and mechanism of torularhodin by investigating the bioavailability of torularhodin in different tissues, the expression of related enzymes and several transporters in the intestine. The results showed that torularhodin entered the intestine faster from micelles (45.21 ± 2.61% was absorbed in the duodenum), and part of it was metabolized into retinol in the anterior segment of the intestine. The expression of genes indicated that absorption of torularhodin in the intestine might require transporter CD36 and SR-B1. The special structure and target organ might be speculated to be the main reason for the low bioavailability of torularhodin in the serum and liver. The results could lay a theoretical foundation for the chemical modification, carrier and subsequent development of torularhodin.

Placental Inositol Reduced in Gestational Diabetes as Glucose Alters Inositol Transporters and IMPA1 Enzyme Expression.

Perturbed inositol physiology in insulin-resistant conditions has led to proposals of inositol supplementation for gestational diabetes (GDM) prevention, but placental inositol biology is poorly understood. Investigate associations of maternal glycemia with placental inositol content, determine glucose effects on placental expression of inositol enzymes and transporters, and examine relations with birthweight. Case-control study of placentae from term singleton pregnancies (GDM n = 24, non-GDM n = 26), and culture of another 9 placentae in different concentrations of glucose and myo-inositol for 48 hours. Placental inositol was quantified by the Megazyme assay. Relative expression of enzymes involved in myo-inositol metabolism and plasma membrane inositol transport was determined by quantitative RT-PCR and immunoblotting. Linear regression analyses were adjusted for maternal age, body mass index, ethnicity, gestational age, and sex. Placental inositol content was 17% lower in GDM compared with non-GDM. Higher maternal mid-gestation glycemia were associated with lower placental inositol. Increasing fasting glycemia was associated with lower protein levels of the myo-inositol synthesis enzyme, IMPA1, and the inositol transporters, SLC5A11 and SLC2A13, the expression of which also correlated with placental inositol content. In vitro, higher glucose concentrations reduced IMPA1 and SLC5A11 mRNA expression. Increasing fasting glycemia positively associated with customized birthweight percentile as expected in cases with low placental inositol, but this association was attenuated with high placental inositol. Glycemia-induced dysregulation of placental inositol synthesis and transport may be implicated in reduced placental inositol content in GDM, and this may in turn be permissive to accelerated fetal growth.

Antidiabetic Effect of Artemether in Db/Db Mice Involves Regulation of AMPK and PI3K/Akt Pathways.

The traditional Chinese medicine has long been used in the treatment of diabetes, one major disease threatening the public health. It has been reported that artemether exerts antidiabetic effects on type 2 diabetes in db/db mice, however the underlying mechanisms remain unknown. In the present study, we show that artemether regulates expression of related enzymes participating in the glucose and lipid metabolism in the liver of db/db mice, which could at least partly explain the improved glucose and lipid metabolism in artemether-treated mice. Additionally, artemether also regulates expression of glycogen synthesis related enzymes in the skeletal muscle of db/db mice, supporting its promotive role in glycogen synthesis. Mechanistically, artemether activates AMPK pathway as well as PI3K/Akt pathway in the liver and skeletal muscle of db/db mice, suggesting that these two signaling pathways are both involved in the antidiabetic effects of artemether on type 2 diabetes in db/db mice. In conclusion, our study connects the antidiabetic effects of artemether to the regulation of metabolic enzymes and signaling pathways, and also provides molecular basis for the potential application of artemether in treating type 2 diabetes.

Exogenous Melatonin Improves Drought Tolerance in Maize Seedlings by Regulating Photosynthesis and the Ascorbate–Glutathione Cycle

Melatonin is known to exert protective effects in maize against drought stress, but the knowledge regarding interaction among the melatonin, photosynthetic efficiency, ascorbate–glutathione cycle still remains elusive. Two contrasting maize (Zea mays L.) genotypes, SD609 (drought-tolerant) and SD902 (drought-sensitive), were subjected to moderate and severe drought stress along with 100 μM melatonin treatment. Exogenous melatonin increased fluorescence curve levels for both genotypes under drought stress, and higher in SD609 than SD902. In both genotypes, melatonin application also significantly increased the photochemical efficiency of photosystem II (PSII), the effective quantum yield of PSII and photosystem I (PSI), and the electron transport rate between PSII and PSI under drought stress, but it decreased the quantum yield of energy dissipation and the donor side and acceptor side impairment of PSI under drought stress, regardless of stress severity. Exogenous melatonin also contributed to the reduction of membrane damage under drought stress, as reflected by significantly decreased levels of MDA, superoxide anions and H2O2 of both genotypes exposed to drought. Furthermore, melatonin treatment increased enzyme activity in AsA–GSH cycle, and upregulated the genes expression of related enzymes in AsA–GSH cycle. The effects of melatonin were more pronounced in SD609 than SD902. Collectively, these results indicated that exogenous melatonin application increased photosynthetic electron transport rate and accelerated the AsA–GSH cycle, which are factors that play an important role in drought tolerance in maize. The beneficial effect of melatonin on the drought-tolerance SD609 was more obvious.

Hot air and UV-C treatments promote anthocyanin accumulation in peach fruit through their regulations of sugars and organic acids

In our present study, we aimed to explore the effects of hot air and UV-C on anthocyanins and the interaction among anthocyanin, sucrose and organic acids in peaches during postharvest storage. Peaches were treated with hot air or UV-C and stored at 1 °C for 35 days. The results showed that both treatments significantly enhanced the accumulation of anthocyanins and suppressed the degradation of sucrose, citric and malic acids. An in vitro test verified that sucrose, citric and malic acid penetrated the tissue and then induced the biosynthesis of anthocyanins by up regulating anthocyanin-related enzymes. In addition, hot air and UV-C directly enhanced the activities and gene expression of related enzymes to promote the accumulation of anthocyanins. PAL, ANS and UFGT played crucial roles in the biosynthesis of anthocyanins in peach fruit after harvest, and these three enzymes can be stimulated by HA, UV-C, sucrose, citric and malic acid.

Exercise-mediated improvement of depression in patients with gastro-esophageal junction cancer is linked to kynurenine metabolism

Background: Exercise may improve depression in cancer patients, yet the molecular mechanism behind this protection is poorly understood. Here, we aimed to explore the link between exercise and regulation of kynurenine (Kyn) metabolism and inflammation in patients with operable gastro-esophageal junction (GEJ) cancer patients, who improved significantly in depression score with exercise training.Material and Methods: Fifty GEJ cancer patients were allocated to 12 weeks of supervised training twice weekly including interval-based aerobic exercise and resistance training, or standard care. Depression score was evaluated by HADS, and blood samples and muscle biopsies were collected for determination of Kyn metabolism and inflammation across the intervention.Results: Depression scores decreased by −1.3 points in the exercise group (p < 0.01), whereas no changes were observed in the control group. Plasma 3-hydroxykynurenine (HK), a Kyn metabolite giving rise to other neurotoxic metabolites, increased by 48% (p <0.001) in the control group, while exercise training attenuated this accumulation. The production of HK is induced by inflammation, and while we observed no differences in systemic pro-inflammatory cytokines, exercise training ameliorated the treatment-induced intramuscular inflammation. Moreover, exercise has been suggested to convert Kyn to the neuroprotective metabolite, kynurenic acid (KA), but despite marked functional and muscular exercise-mediated adaptations, we did not observe any enhancement of KA production and related enzyme expression in the muscles of GEJ cancer patients.Conclusion: Exercise training reduced symptoms of depression in patients with GEJ cancer, and this effect was associated with an exercise-dependent attenuation of the inflammation-induced conversion of Kyn to neurotoxic metabolites.

Lithium chloride inhibits StAR and progesterone production through GSK-3β and ERK1/2 signaling pathways in human granulosa-lutein cells

Lithium chloride (LiCl) is a widely-used medication to treat neurological disorders that has undesirable side effects on the female reproductive system. It has been show that LiCl can inhibit ovarian folliculogenesis, promote follicle atresia and suppress steroid hormone production in rodents. However, the effects of LiCl on human ovarian steroidogenesis remain completely unknown. In this study, both primary and immortalized human granulosa-lutein (hGL) cells were used to investigate the effects of LiCl on progesterone production and its related enzyme expression as well as the underlying mechanisms. Our results showed that LiCl significantly down-regulated the steroidogenic acute regulatory protein (StAR) expression and subsequent progesterone production in hGL cells. Additionally, LiCl induced the phosphorylation of GSK-3β and ERK1/2 but not AKT or CREB. Knockdown of endogenous GSK-3β or inhibition of ERK1/2 partially reversed LiCl-induced down-regulation of StAR. Furthermore, by using dual inhibition approaches, the results showed that both GSK-3β and ERK1/2 signaling mediated the regulatory effect of LiCl on StAR expression. Our findings deepen our understanding of the pathological effects and the underlying molecular mechanisms of how lithium might affect the female reproductive system.

Hexosamine biosynthesis in keratinocytes: roles of GFAT and GNPDA enzymes in the maintenance of UDP-GlcNAc content and hyaluronan synthesis.

UDP-N-acetylglucosamine (UDP-GlcNAc) is a glucose metabolite with pivotal functions as a key substrate for the synthesis of glycoconjugates like hyaluronan, and as a metabolic sensor that controls cell functions through O-GlcNAc modification of intracellular proteins. However, little is known about the regulation of hexosamine biosynthesis that controls UDP-GlcNAc content. Four enzymes can catalyze the crucial starting point of the pathway, conversion of fructose-6-phosphate (Fru6P) to glucosamine-6-phosphate (GlcN6P): glutamine-fructose-6-phosphate aminotransferases (GFAT1 and 2) and glucosamine-6-phosphate deaminases (GNPDA1 and 2). Using siRNA silencing, we studied the contributions of these enzymes to UDP-GlcNAc content and hyaluronan synthesis in human keratinocytes. Depletion of GFAT1 reduced the cellular pool of UDP-GlcNAc and hyaluronan synthesis, while simultaneous blocking of both GNPDA1 and GDPDA2 exerted opposite effects, indicating that in standard culture conditions keratinocyte GNPDAs mainly catalyzed the reaction from GlcN6P back to Fru6P. However, when hexosamine biosynthesis was blocked by GFAT1 siRNA, the effect by GNPDAs was reversed, now catalyzing Fru6P towards GlcN6P, likely in an attempt to maintain UDP-GlcNAc content. Silencing of these enzymes also changed the gene expression of related enzymes: GNPDA1 siRNA induced GFAT2 which was hardly measurable in these cells under standard culture conditions, GNPDA2 siRNA increased GFAT1, and GFAT1 siRNA increased the expression of hyaluronan synthase 2 (HAS2). Silencing of GFAT1 stimulated GNPDA1 and GDPDA2, and inhibited cell migration. The multiple delicate adjustments of these reactions demonstrate the importance of hexosamine biosynthesis in cellular homeostasis, known to be deranged in diseases like diabetes and cancer.

Deprivation of L-Arginine Induces Oxidative Stress Mediated Apoptosis in Leishmania donovani Promastigotes: Contribution of the Polyamine Pathway

The growth and survival of intracellular parasites depends on the availability of extracellular nutrients. Deprivation of nutrients viz glucose or amino acid alters redox balance in mammalian cells as well as some lower organisms. To further understand the relationship, the mechanistic role of L-arginine in regulation of redox mediated survival of Leishmania donovani promastigotes was investigated. L-arginine deprivation from the culture medium was found to inhibit cell growth, reduce proliferation and increase L-arginine uptake. Relative expression of enzymes, involved in L-arginine metabolism, which leads to polyamine and trypanothione biosynthesis, were downregulated causing decreased production of polyamines in L-arginine deprived parasites and cell death. The resultant increase in reactive oxygen species (ROS), due to L-arginine deprivation, correlated with increased NADP+/NADPH ratio, decreased superoxide dismutase (SOD) level, increased lipid peroxidation and reduced thiol content. A deficiency of L-arginine triggered phosphatidyl serine externalization, a change in mitochondrial membrane potential, release of intracellular calcium and cytochrome-c. This finally led to DNA damage in Leishmania promastigotes. In summary, the growth and survival of Leishmania depends on the availability of extracellular L-arginine. In its absence the parasite undergoes ROS mediated, caspase-independent apoptosis-like cell death. Therefore, L-arginine metabolism pathway could be a probable target for controlling the growth of Leishmania parasites and disease pathogenesis.

Quantitative Gene Expression of Peroxidase, Polyphenoloxidase and Catalase as Molecular Markers for Resistance against &amp;lt;i&amp;gt;Ralstonia solanacearum&amp;lt;/i&amp;gt;

Brown rot or bacterial wilt of potato caused by Ralstonia solanacearum, is an economically important disease. Potato, cv. Nicola, was found to be relatively highly resistant to the infection with R. solanacearum and showed 15.12% wilt disease index, meantime, cv. Kara showed intermediate resistance with 37.40% disease index while, cv. Spunta was susceptible and showed 80.33% disease index. The role of defense-related enzymes in imparting resistance in potato against R. solanacearum was investigated by quantifying enzymes activity and gene expression of three defense- related enzymes, peroxidase, polyphenol oxidase and catalase. Peroxidase showed maximum activity 0.488 min-1&middotg-1 early at 12 h after pathogen inoculation in the cv. Nicola, whereas in susceptible cultivar Spunta showed lower activity of maximum 0.226 min-1&middotg-1 later at 48 h after inoculation. While, the moderately resistant cultivar Kara showed intermediate activity for the peak and its time. Meanwhile, polyphenol oxidase showed similar trends to that of peroxidase. On the contrary, catalase showed the highest activity values in the susceptible, cv. Spunta, while, in relatively highly resistant (cv. Nicola) and the moderately resistant (cv. Kara) showed lower values of activity and up to 96 h after inoculation. Meanwhile, gene expression of related enzymes the RT-PCR was used. At zero time, the relatively highly resistant potato cultivar, Nicola, showed the highest values of gene expression for both Peroxidase (POD) and Poly Phenol Oxidase (PPO). While, the susceptible potato cultivar, Spunta showed the lowest values. On the contrary, Catalase (CAT) gene expression was the highest in the susceptible, cv. Spunta, and was the lowest in the relatively highly resistant, cv. Nicola, while, was of intermediate values in the intermediate resistance, cv. Kara. Results show that peroxidase and polyphenol oxidase activities can be used as biochemical markers to reveal the resistance and susceptibility nature of potato cultivars against bacterial wilt disease of potato caused by R. solanacaerum.

Targeted Integration of RNA-Seq and Metabolite Data to Elucidate Curcuminoid Biosynthesis in Four Curcuma Species.

Curcuminoids, namely curcumin and its analogs, are secondary metabolites that act as the primary active constituents of turmeric (Curcuma longa). The contents of these curcuminoids vary among species in the genus Curcuma. For this reason, we compared two wild strains and two cultivars to understand the differences in the synthesis of curcuminoids. Because the fluxes of metabolic reactions depend on the amounts of their substrate and the activity of the catalysts, we analyzed the metabolite concentrations and gene expression of related enzymes. We developed a method based on RNA sequencing (RNA-Seq) analysis that focuses on a specific set of genes to detect expression differences between species in detail. We developed a 'selection-first' method for RNA-Seq analysis in which short reads are mapped to selected enzymes in the target biosynthetic pathways in order to reduce the effect of mapping errors. Using this method, we found that the difference in the contents of curcuminoids among the species, as measured by gas chromatography-mass spectrometry, could be explained by the changes in the expression of genes encoding diketide-CoA synthase, and curcumin synthase at the branching point of the curcuminoid biosynthesis pathway.

Effects of zinc deficiency and zinc supplementation on homocysteine levels and related enzyme expression in rats

Methionine synthase (MS) and betaine-homocysteine methyltransferase (BHMT) are both zinc (Zn)-dependent methyltransferases and involved in the methylation of homocysteine. The objective of this study was to investigate the effects of dietary Zn supply on homocysteine levels and expression of the two enzymes in growing rats. Male weanling Sprague-Dawley rats were assigned randomly to four dietary groups (n=8/group) for 3 weeks: Zn deficient (ZD; <1mg Zn/kg); Zn control (ZC; 30mg Zn/kg); Zn supplemented (ZS; 300mg Zn/kg); pair fed (PF; 30mg Zn/kg) to the ZD group. Serum and femur Zn concentrations were 83% and 58% lower in ZD, and 49% and 62% higher in ZS compared to ZC (P<0.001), respectively. The ZD rats had lower feed intake (37%), body weight gains (45%), liver (43%) and kidney (31%) weights than those of ZC (P<0.001), but these parameters in ZD were not significantly different from the PF controls. Serum homocysteine concentrations were 65% higher in ZD compared to PF (P<0.05), and there was no significant difference in serum folate levels between ZD and PF groups. The mRNA expression of liver and kidney MS was 57% and 38% lower in ZD than PF (P<0.001), respectively. Hepatic and renal BHMT mRNA levels were not altered in ZD compared to controls. The aforementioned measurements were not significantly different between ZS and ZC groups, except Zn levels. These results demonstrated that homocysteine homeostasis appeared to be disturbed by Zn deficiency but not Zn supplementation, and elevated serum homocysteine might be due to reduced expression of MS during Zn deficiency.

1,25-Dihydroxyvitamin D regulation of glucose metabolism in Harvey-ras transformed MCF10A human breast epithelial cells

This study was designed to investigate the impact of 1,25-dihydroxyvitamin D (1,25(OH)2D) on glucose metabolism during early cancer progression. Untransformed and ras-oncogene transfected (ras) MCF10A human breast epithelial cells were employed to model early breast cancer progression. 1,25(OH)2D modified the response of the ras cells to glucose restriction, suggesting 1,25(OH)2D may reduce the ras cell glucose addiction noted in cancer cells. To understand the 1,25(OH)2D regulation of glucose metabolism, following four-day 1,25(OH)2D treatment, metabolite fluxes at the cell membrane were measured by a nanoprobe biosensor, [13C6]glucose flux by 13C-mass isotopomer distribution analysis of media metabolites, intracellular metabolite levels by NMR, and gene expression of related enzymes was assessed. Treatment with 1,25(OH)2D reduced glycolysis as flux of glucose to 3-phosphoglycerate was reduced by 15% (P=0.017) and 32% (P<0.003) in MCF10A and ras cells respectively. In the ras cells, 1,25(OH)2D reduced lactate dehydrogenase activity by 15% (P<0.05) with a concomitant 10% reduction in the flux of glucose to lactate (P=0.006), and reduction in the level of intracellular lactate by 55% (P=0.029). Treatment with 1,25(OH)2D reduced flux of glucose to acetyl-coA 24% (P=0.002) and 41% (P<0.001), and flux to oxaloacetate 33% (P=0.003) and 34% (P=0.027) in the MCF10A and ras cells, respectively, suggesting a reduction in tricarboxylic acid (TCA) cycle activity. The results suggest a novel mechanism involving the regulation of glucose metabolism by which 1,25(OH)2D may prevent breast cancer progression.

Dietary effect of royal jelly supplementation on epidermal levels of hydration, filaggrins, free amino acids and the related enzyme expression in UV irradiated hairless mice

Dietary effect of royal jelly supplementation on epidermal levels of hydration, filaggrins, free amino acids and the related enzyme expression in UV irradiated hairless mice