Transketolase promotes MAFLD by limiting inosine-induced mitochondrial activity.

Metabolic dysfunction-associated fatty liver disease (MAFLD) has a global prevalence of about 25% and no approved therapy. Using metabolomic and proteomic analyses, we identified high expression of hepatic transketolase (TKT), a metabolic enzyme of the pentose phosphate pathway, in human and mouse MAFLD. Hyperinsulinemia promoted TKT expression through the insulin receptor-CCAAT/enhancer-binding protein alpha axis. Utilizing liver-specific TKT overexpression and knockout mouse models, we demonstrated that TKT was sufficient and required for MAFLD progression. Further metabolic flux analysis revealed that Tkt deletion increased hepatic inosine levels to activate the protein kinase A-cAMP response element binding protein cascade, promote phosphatidylcholine synthesis, and improve mitochondrial function. Moreover, insulin induced hepatic TKT to limit inosine-dependent mitochondrial activity. Importantly, N-acetylgalactosamine (GalNAc)-siRNA conjugates targeting hepatic TKT showed promising therapeutic effects on mouse MAFLD. Our study uncovers how hyperinsulinemia regulates TKT-orchestrated inosine metabolism and mitochondrial function and provides a novel therapeutic strategy for MAFLD prevention and treatment.

Loss of transketolase promotes the anti-diabetic role of brown adipose tissues.

Transketolase (TKT), an enzyme in the non-oxidative branch of the pentose phosphate pathway (PPP), bi-directionally regulates the carbon flux between the PPP and glycolysis. Loss of TKT in adipose tissues decreased glycolysis and increased lipolysis and uncoupling protein-1 (UCP1) expression, protecting mice from high-fat diet-induced obesity. However, the role of TKT in brown adipose tissue (BAT)-dependent glucose homeostasis under normal chow diet remains to be elucidated. We found that TKT ablation increased levels of glucose transporter 4 (GLUT4), promoting glucose uptake and glycogen accumulation in BAT. Using the streptozotocin (STZ)-induced diabetic mouse model, we discovered that enhanced glucose uptake due to TKT deficiency in BAT contributed to decreasing blood glucose and weight loss, protecting mice from STZ-induced diabetes. Mechanistically, TKT deficiency decreased the level of thioredoxin-interacting protein, a known inhibitor for GLUT4, by decreasing NADPH and glutathione levels and inducing oxidative stress in BAT. Therefore, our data reveal a new role of TKT in regulating the anti-diabetic function of BAT as well as glucose homeostasis.

MEK1-dependent MondoA phosphorylation regulates glucose uptake in response to ketone bodies in colorectal cancer cells.

The Mondo family transcription factor MondoA plays a pivotal role in sensing metabolites, such as glucose, glutamine, and lactic acid, to regulate glucose metabolism and cell proliferation. Ketone bodies are important signals for reducing glucose uptake. However, it is unclear whether MondoA functions in ketone body-regulated glucose transport. Here we reported that ketone bodies promoted MondoA nuclear translocation and binding to the promoter of its target gene TXNIP. Ketone bodies reduced glucose uptake, increased apoptosis and decreased proliferation of colorectal cancer cells, which was impeded by MondoA knockdown. Moreover, we identified MEK1 as a novel component of the MondoA protein complex using a proteomic approach. Mechanistically, MEK1 interacted with MondoA and enhanced tyrosine 222, but not serine or threonine, phosphorylation of MondoA, inhibiting MondoA nuclear translocation and transcriptional activity. Ketone bodies decreased MEK1-dependent MondoA phosphorylation by blocking MondoA and MEK1 interaction, leading to MondoA nuclear translocation, TXNIP transcription, and inhibition of glucose uptake. Therefore, our study not only demonstrated that ketone bodies reduce glucose uptake, promote apoptosis, and inhibit cell proliferation in colorectal cancer cells by regulating MondoA phosphorylation but also identified MEK1-dependent phosphorylation as a new mechanism to manipulate MondoA activity.

Non-oxidative pentose phosphate pathway controls regulatory T cell function by integrating metabolism and epigenetics.

Regulatory T (Treg) cells are critical for maintaining immune homeostasis and preventing autoimmunity. Here, we show that the non-oxidative pentose phosphate pathway (PPP) regulates Treg function to prevent autoimmunity. Deletion of transketolase (TKT), an indispensable enzyme of non-oxidative PPP, in Treg cells causes a fatal autoimmune disease in mice, with impaired Treg suppressive capability despite regular Treg numbers and normal Foxp3 expression levels. Mechanistically, reduced glycolysis and enhanced oxidative stress induced by TKT deficiency triggers excessive fatty acid and amino acid catabolism, resulting in uncontrolled oxidative phosphorylation and impaired mitochondrial fitness. Reduced α-KG levels as a result of reductive TCA cycle activity leads to DNA hypermethylation, thereby limiting functional gene expression and suppressive activity of TKT-deficient Treg cells. We also find that TKT levels are frequently downregulated in Treg cells of people with autoimmune disorders. Our study identifies the non-oxidative PPP as an integrator of metabolic and epigenetic processes that control Treg function.

TKT maintains intestinal ATP production and inhibits apoptosis-induced colitis.

Inflammatory bowel disease (IBD) has a close association with transketolase (TKT) that links glycolysis and the pentose phosphate pathway (PPP). However, how TKT functions in the intestinal epithelium remains to be elucidated. To address this question, we specifically delete TKT in intestinal epithelial cells (IECs). IEC TKT-deficient mice are growth retarded and suffer from spontaneous colitis. TKT ablation brings about striking alterations of the intestine, including extensive mucosal erosion, aberrant tight junctions, impaired barrier function, and increased inflammatory cell infiltration. Mechanistically, TKT deficiency significantly accumulates PPP metabolites and decreases glycolytic metabolites, thereby reducing ATP production, which results in excessive apoptosis and defective intestinal barrier. Therefore, our data demonstrate that TKT serves as an essential guardian of intestinal integrity and barrier function as well as a potential therapeutic target for intestinal disorders.

MondoA-Thioredoxin-Interacting Protein Axis Maintains Regulatory T-Cell Identity and Function in Colorectal Cancer Microenvironment.

BACKGROUND & AIMS: The metabolic features and function of intratumoral regulatory T cells (Tregs) are ambiguous in colorectal cancer. Tumor-infiltrating Tregs are reprogrammed to exhibit high glucose-depleting properties and adapt to the glucose-restricted microenvironment. The glucose-responsive transcription factor MondoA is highly expressed in Tregs. However, the role of MondoA in colorectal cancer-infiltrating Tregs in response to glucose limitation remains to be elucidated. METHODS: We performed studies using mice, in which MondoA was conditionally deleted in Tregs, and human colorectal cancer tissues. Seahorse and other metabolic assays were used to assess Treg metabolism. To study the role of Tregs in antitumor immunity, we used a subcutaneous MC38 colorectal cancer model and induced colitis-associated colorectal cancer in mice by azoxymethane and dextran sodium sulfate. RESULTS: Our analysis of single-cell RNA sequencing data of patients with colorectal cancer revealed that intratumoral Tregs featured low activity of the MondoA-thioredoxin-interacting protein (TXNIP) axis and increased glucose uptake. Although MondoA-deficient Tregs were less immune suppressive and selectively promoted T-helper (Th) cell type 1 (Th1) responses in a subcutaneous MC38 tumor model, Treg-specific MondoA knockout mice were more susceptible to azoxymethane-DSS-induced colorectal cancer. Mechanistically, suppression of the MondoA-TXNIP axis promoted glucose uptake and glycolysis, induced hyperglycolytic Th17-like Tregs, which facilitated Th17 inflammation, promoted interleukin 17A-induced of CD8+ T-cell exhaustion, and drove colorectal carcinogenesis. Blockade of interleukin 17A reduced tumor progression and minimized the susceptibility of MondoA-deficient mice to colorectal carcinogenesis. CONCLUSIONS: The MondoA-TXNIP axis is a critical metabolic regulator of Treg identity and function in the colorectal cancer microenvironment and a promising target for cancer therapy.

ERα down-regulates carbohydrate responsive element binding protein and decreases aerobic glycolysis in liver cancer cells.

Deregulated metabolism is one of the characteristics of hepatocellular carcinoma. Sex hormone receptor signalling has been involved in the marked gender dimorphism of hepatocellular carcinoma pathogenesis. Oestrogen receptor (ER) has been reported to reduce the incidence of liver cancer. However, it remains unclear how oestrogen and ER regulate metabolic alterations in liver tumour cells. Our previous work revealed that ERα interacted with carbohydrate responsive element binding protein (ChREBP), which is a transcription factor promoting aerobic glycolysis and proliferation of hepatoma cells. Here, the data showed that ERα overexpression with E2 treatment reduced aerobic glycolysis and cell proliferation of hepatoma cells. In addition to modestly down-regulating ChREBP transcription, ERα promoted ChREBP degradation. ERα co-immunoprecipitated with both ChREBP-α and ChREBP-ß, the two known subtypes of ChREBP. Although E2 promoted ERα to translocate to the nucleus, it did not change subcellular localization of ChREBP. In addition to interacting with ChREBP-ß and promoting its degradation, ERα decreased ChREBP-α-induced ChREBP-ß transcription. Taken together, we confirmed an original role of ERα in suppressing aerobic glycolysis in liver cancer cells and elucidated the mechanism by which ERα and ChREBP-α together regulated ChREBP-ß expression.

Prediction of net energy values in expeller-pressed and solvent-extracted rapeseed meal for growing pigs.

OBJECTIVE: The objective of this study was to determine net energy (NE) of expeller-press (EP-RSM) and solvent-extracted rapeseed meal (SE-RSM) and to establish equations for predicting the NE in rapeseed meal (RSM) fed to growing pigs. METHODS: Thirty-six barrows (initial body weight [BW], 41.1±2.2 kg) were allotted into 6 diets comprising a corn-soybean meal basal diet and 5 diets containing 19.50% RSM added at the expense of corn and soybean meal. The experiment had 6 periods and 6 replicate pigs per diet. During each period, the pigs were individually housed in metabolism crates for 16 days which included 7 days for adaption to diets. On day 8, pigs were transferred to respiration chambers and fed their respective diet at 2,000 kJ metabolizable energy (ME)/kg BW0.6/d. Feces and urine were collected, and daily heat production was measured from day 9 to 13. On days 14 and 15, the pigs were fed at 890 kJ ME/kg BW0.6/d and fasted on day 16 for evaluation of fasting heat production (FHP). RESULTS: The FHP of pigs averaged 790 kJ/kg BW0.6/d and was not affected by the diet composition. The NE values were 10.80 and 8.45 MJ/kg DM for EP-RSM and SE-RSM, respectively. The NE value was positively correlated with gross energy (GE), digestible energy (DE), ME, and ether extract (EE). The best fit equation for NE of RSM was NE (MJ/kg DM) = 1.14×DE (MJ/kg DM)+0.46×crude protein (% of DM)-25.24 (n = 8, R2 = 0.96, p<0.01). The equation NE (MJ/kg DM) = 0.22×EE (% of DM)-0.79×ash (% of DM)+14.36 (n = 8, R2 = 0.77, p = 0.018) may be utilized to quickly determine the NE in RSM when DE or ME values are unavailable. CONCLUSION: The NE values of EP-RSM and SE-RSM were 10.80 and 8.45 MJ/kg DM. The NE value of RSM can be well predicted based on energy content (GE, DE, and ME) and proximate analysis.

The Role of the Pentose Phosphate Pathway in Diabetes and Cancer.

The pentose phosphate pathway (PPP) branches from glucose 6-phosphate (G6P), produces NADPH and ribose 5-phosphate (R5P), and shunts carbons back to the glycolytic or gluconeogenic pathway. The PPP has been demonstrated to be a major regulator for cellular reduction-oxidation (redox) homeostasis and biosynthesis. Enzymes in the PPP are reported to play important roles in many human diseases. In this review, we will discuss the role of the PPP in type 2 diabetes and cancer.

Transketolase Deficiency in Adipose Tissues Protects Mice From Diet-Induced Obesity by Promoting Lipolysis.

Obesity has recently become a prevalent health threat worldwide. Although emerging evidence has suggested a strong link between the pentose phosphate pathway (PPP) and obesity, the role of transketolase (TKT), an enzyme in the nonoxidative branch of the PPP that connects PPP and glycolysis, remains obscure in adipose tissues. In this study, we specifically deleted TKT in mouse adipocytes and found no obvious phenotype upon normal diet feeding. However, adipocyte TKT abrogation attenuated high-fat diet-induced obesity, reduced hepatic steatosis, improved glucose tolerance, alleviated insulin resistance, and increased energy expenditure. Mechanistically, TKT deficiency accumulated nonoxidative PPP metabolites and decreased glycolysis and pyruvate input into the mitochondria, leading to increased lipolytic enzyme gene expression and enhanced lipolysis, fatty acid oxidation, and mitochondrial respiration. Therefore, our data not only identify a novel role of TKT in regulating lipolysis and obesity but also suggest that limiting glucose-derived carbon into the mitochondria induces lipid catabolism and energy expenditure.

The ubiquitination ligase SMURF2 reduces aerobic glycolysis and colorectal cancer cell proliferation by promoting ChREBP ubiquitination and degradation.

The glucose-responsive transcription factor carbohydrate response element-binding protein (ChREBP) critically promotes aerobic glycolysis and cell proliferation in colorectal cancer cells. It has been reported that ubiquitination may be important in the regulation of ChREBP protein levels and activities. However, the ChREBP-specific E3 ligase and molecular mechanism of ChREBP ubiquitination remains unclear. Using database exploration and expression analysis, we found here that levels of the E3 ligase SMURF2 (Smad-ubiquitination regulatory factor 2) negatively correlate with those of ChREBP in cancer tissues and cell lines. We observed that SMURF2 interacts with ChREBP and promotes ChREBP ubiquitination and degradation via the proteasome pathway. Interestingly, ectopic SMURF2 expression not only decreased ChREBP levels but also reduced aerobic glycolysis, increased oxygen consumption, and decreased cell proliferation in colorectal cancer cells. Moreover, SMURF2 knockdown increased aerobic glycolysis, decreased oxygen consumption, and enhanced cell proliferation in these cells, mostly because of increased ChREBP accumulation. Furthermore, we identified Ser/Thr kinase AKT as an upstream suppressor of SMURF2 that protects ChREBP from ubiquitin-mediated degradation. Taken together, our results indicate that SMURF2 reduces aerobic glycolysis and cell proliferation by promoting ChREBP ubiquitination and degradation via the proteasome pathway in colorectal cancer cells. We conclude that the SMURF2-ChREBP interaction might represent a potential target for managing colorectal cancer.

Transketolase Deficiency Protects the Liver from DNA Damage by Increasing Levels of Ribose 5-Phosphate and Nucleotides.

De novo nucleotide biosynthesis is essential for maintaining cellular nucleotide pools, the suppression of which leads to genome instability. The metabolic enzyme transketolase (TKT) in the nonoxidative branch of the pentose phosphate pathway (PPP) regulates ribose 5-phosphate (R5P) levels and de novo nucleotide biosynthesis. TKT is required for maintaining cell proliferation in human liver cancer cell lines, yet the role of TKT in liver injury and cancer initiation remains to be elucidated. In this study, we generated a liver-specific TKT knockout mouse strain by crossing TKTflox/flox mice with albumin-Cre mice. Loss of TKT in hepatocytes protected the liver from diethylnitrosamine (DEN)-induced DNA damage without altering DEN metabolism. DEN treatment of TKT-null liver increased levels of R5P and promoted de novo nucleotide synthesis. More importantly, supplementation of dNTPs in primary hepatocytes alleviated DEN-induced DNA damage, cell death, inflammatory response, and cell proliferation. Furthermore, DEN and high-fat diet (HFD)-induced liver carcinogenesis was reduced in TKTflox/floxAlb-Cre mice compared with control littermates. Mechanistically, loss of TKT in the liver increased apoptosis, reduced cell proliferation, decreased TNFα, IL6, and STAT3 levels, and alleviated DEN/HFD-induced hepatic steatosis and fibrosis. Together, our data identify a key role for TKT in promoting genome instability during liver injury and tumor initiation. SIGNIFICANCE: These findings identify transketolase as a novel metabolic target to maintain genome stability and reduce liver carcinogenesis.

Metabolizable energy requirement for maintenance estimated by regression analysis of body weight gain or metabolizable energy intake in growing pigs

Objective: Feed energy required for pigs is first prioritized to meet maintenance costs. Additional energy intake in excess of the energy requirement for maintenance is retained as protein and fat in the body, leading to weight gain. The objective of this study was to estimate the ME requirements for maintenance (MEm) by regressing body weight gain against ME intake (MEI) in growing pigs. Methods: Thirty-six growing pigs (26.3 ± 1.7 kg) were allotted to 1 of 6 treatments with 6 replicates per treatment in a randomized complete block design. Treatments were 6 feeding levels which were calculated as 50, 60, 70, 80, 90 or 100% of the estimated ad libitum MEI (2,400 kJ/kg BW0.60·d). All pigs were individually housed in metabolism crates for 30 d and weighed every 5 d. Moreover, each pig from each treatment was placed in the open-circuit respiration chambers to measure heat production (HP) and energy retained as protein (REp) and fat (REf) every 5 d. Serum biochemical parameters of pigs were analyzed at the end of the experiment. Results: The ADG and HP as well as the REp and REf linearly increased with increasing feed intake (p < 0.010). ß-hydroxybutyrate (BHBA) concentration of serum tended to increase with increasing feed intake (p = 0.080). The regression equations of MEI on ADG were MEI, kJ/kg BW0.60·d = 1.88 × ADG, g/d + 782 (R2=0.86) and MEm was estimated at 782 kJ/kg BW0.60·d. Protein retention of growing pigs would be positive while REf would be negative at this feeding level via regression equations of REp and REf on MEI. Conclusion: The MEm was estimated at 782 kJ/kg BW0.60·d in current experiment. Furthermore, growing pigs will deposit protein and oxidize fat if provided feed at the estimated maintenance level.

Methodologies on estimating the energy requirements for maintenance and determining the net energy contents of feed ingredients in swine: a review of recent work.

In the past two decades, a considerable amount of research has focused on the determination of the digestible (DE) and metabolizable energy (ME) contents of feed ingredients fed to swine. Compared with the DE and ME systems, the net energy (NE) system is assumed to be the most accurate estimate of the energy actually available to the animal. However, published data pertaining to the measured NE content of ingredients fed to growing pigs are limited. Therefore, the Feed Data Group at the Ministry of Agricultural Feed Industry Centre (MAFIC) located at China Agricultural University has evaluated the NE content of many ingredients using indirect calorimetry. The present review summarizes the NE research works conducted at MAFIC and compares these results with those from other research groups on methodological aspect. These research projects mainly focus on estimating the energy requirements for maintenance and its impact on the determination, prediction, and validation of the NE content of several ingredients fed to swine. The estimation of maintenance energy is affected by methodology, growth stage, and previous feeding level. The fasting heat production method and the curvilinear regression method were used in MAFIC to estimate the NE requirement for maintenance. The NE contents of different feedstuffs were determined using indirect calorimetry through standard experimental procedure in MAFIC. Previously generated NE equations can also be used to predict NE in situations where calorimeters are not available. Although popular, the caloric efficiency is not a generally accepted method to validate the energy content of individual feedstuffs. In the future, more accurate and dynamic NE prediction equations aiming at specific ingredients should be established, and more practical validation approaches need to be developed.

Net energy content of rice bran, defatted rice bran, corn gluten feed, and corn germ meal fed to growing pigs using indirect calorimetry.

The objective of this experiment was to determine the effects of increased fiber content in diets on heat production (HP) and NE:ME ratio and to determine the NE content and NE:ME ratio of full-fat rice bran (FFRB), defatted rice bran (DFRB), corn gluten feed (CGF), and corn germ meal (CGM) fed to growing barrows using indirect calorimetry (IC). Thirty growing barrows (28.5 ± 2.4 kg BW) were allotted in a completely randomized design to 5 dietary treatments that included a corn-soybean meal basal diet and 4 experimental diets with a constant ratio of corn and soybean meal (difference method) containing 30% FFRB, DFRB, CGF, and CGF. Pigs were housed in individual metabolism crates for 20 d including 14-d adaptation to the diet and 6 d to determine the HP and total collection of feces and urine in respiration chambers. Pigs were fed their respective diets at 550 kcal ME·kg BW0.60-1·d-1 on the basis of BW measured on days 0, 7, and 14. The apparent total tract digestibility (ATTD) of DM, GE, and OM were greater (P < 0.01) in pigs fed the basal diet. The ATTD of DM, GE, and OM in pigs fed the DFRB diet were lesser (P < 0.01) when compared with those fed the basal and FFRB diets. The ATTD of ether extract (EE) in pigs fed the FFRB diet was greater (P < 0.01) compared with those fed basal, DFRB, CGF, and CGM diets. The HP adjusted for the same ME intake was greater (P < 0.01) in pigs fed the DFRB, CGF, and CGM diets compared with those fed basal and FFRB diets. The NE:ME ratio in pigs fed the FFRB diet was greater (P < 0.01) when compared with those fed the DFRB, CGF, and CGM diets. The NE content of FFRB, DFRB, CGF, and CGM determined using the IC method were 2,952, 1,100, 1,747, and 2,079 kcal/kg DM, respectively. The NE content of FFRB, CGF, and CGM determined using the IC method were 3.5%, 3.8%, and 1.8% greater, respectively, than the predicted values, whereas NE content of DFRB determined using the IC method was 2.1% lower than the predicted values. In conclusion, pigs fed the fiber-rich ingredients had greater HP and lower nutrient digestibility. However, pigs fed FFRB diets containing greater fat content had a lower heat increment and, therefore, higher utilization efficiency. The NE:ME ratio ranged from 71.6% to 82.4%. The NE of FFRB, DFRB, CGF, and CGM determined using the IC method were 2,952, 1,100, 1,747, and 2,079 kcal/kg DM, respectively.

Determination of net energy content of dietary lipids fed to growing pigs using indirect calorimetry.

The objective of this experiment was to determine the NE content of different dietary lipids fed to growing pigs using indirect calorimetry. Thirty-six growing (initial BW: 41.1 ± 3.1 kg) barrows were allotted to 6 diets based on completely randomized design with 6 replicate pigs per diet. Diets included a corn-soybean meal basal diet and 5 test diets each containing 10% palm oil, poultry fat, fish oil, corn oil, or flaxseed oil at the expense of corn and soybean meal. During each period, pigs were individually housed in metabolism crates for 14 d, which included 7 d for adaptation to feed, metabolism crates, and environmental conditions. On day 8, pigs were transferred to the open-circuit respiration chambers and fed 1 of the 6 diets at 2.3 MJ ME/kg BW0.6/day. Total feces and urine were collected and daily heat production (HP) was also calculated from day 9 to day 13. On the last day of each period (day 14), pigs were fasted and the fasting heat production (FHP) was measured. The results show that the FHP of pigs averaged 809 kJ/kg BW0.6·day-1 and was not affected by diet characteristics. The DE values were 35.98, 36.84, 37.11, 38.95, and 38.38 MJ/kg DM, the ME values were 35.79, 36.56, 36.92, 37.73, and 38.11 MJ/kg DM, and the NE values were 32.42, 33.21, 33.77, 34.00, and 34.12 MJ/kg DM, for the palm oil, poultry fat, fish oil, corn oil, and flaxseed oil, respectively. Based on our result, we concluded that the DE content of dietary lipid varied from 91% to 98% of its GE content, the ME content of dietary lipid was approximately 99% of its DE content, and the NE content of dietary lipid was approximately 90% of its ME content in growing pigs.

Net energy content of rice bran, corn germ meal, corn gluten feed, peanut meal, and sunflower meal in growing pigs.

OBJECTIVE: The objective of this experiment was to determine the net energy (NE) content of full-fat rice bran (FFRB), corn germ meal (CGM), corn gluten feed (CGF), solvent-extracted peanut meal (PNM), and dehulled sunflower meal (SFM) fed to growing pigs using indirect calorimetry or published prediction equations. METHODS: Twelve growing barrows with an average initial body weight (BW) of 32.4±3.3 kg were allotted to a replicated 3×6 Youden square design with 3 successive periods and 6 diets. During each period, pigs were individually housed in metabolism crates for 16 d, which included 7 days for adaptation. On d 8, the pigs were transferred to the respiration chambers and fed one of the 6 diets at 2.0 MJ metabolizable energy (ME)/kg BW0.6/d. Total feces and urine were collected and daily heat production was measured from d 9 to d 13. On d 14 and d15, pigs were fed at their maintenance energy requirement level. On the last day pigs were fasted and fasting heat production was measured. RESULTS: The NE of FFRB, CGM, CGF, PNM, and SFM measured by indirect calorimetry method was 12.33, 8.75, 7.51, 10.79, and 6.49 MJ/kg dry matter (DM), respectively. The NE/ME ratios ranged from 67.2% (SFM) to 78.5% (CGF). The NE values for the 5 ingredients calculated according to the prediction equations were 12.22, 8.55, 6.79, 10.51, and 6.17 MJ/kg DM, respectively. CONCLUSION: The NE values were the highest for FFRB and PNM and the lowest in the corn co-products and SFM. The average NE of the 5 ingredients measured by indirect calorimetry method in the current study was greater than values predicted from NE prediction equations (0.32 MJ/kg DM).

Determination of net energy content of soybean oil fed to growing pigs using indirect calorimetry.

The objectives of this experiment were: (i) to determine the net energy (NE) of soybean oil (SBO) fed to growing pigs using indirect calorimetry (IC); and (ii) to evaluate the effects of inclusion rate of SBO on heat production, oxidative status and nutrient digestibility in growing pigs. Eighteen growing barrows were allotted to three diets based on completely randomized design with six replicate pigs (period) per diet. Diets included a corn-soybean meal basal diet and two test diets containing 5% or 10% SBO at the expense of corn and soybean meal. During each period, pigs were individually housed in metabolism crates for 14 days, including 7 days to adapt to feed, metabolism crate and environmental conditions. On day 8, pigs were transferred to the open-circuit respiration chambers for measurement of daily O2 consumption and CO2 and CH4 production. During this time, pigs were fed one of the three diets at 2.4 MJ metabolizable energy/kg body weight (BW)0.6 /day. Total feces and urine were collected and daily total heat production (THP) was measured from days 9 to 13 and fasted on day 14 to evaluate their fasting heat production (FHP). The results show that trends of decreased apparent total tract digestibility of neutral detergent fiber (linear, P = 0.09) and acid detergent fiber (linear, P = 0.07) were observed as the content of dietary lipids increased. The average THP for the three diets were 1326, 1208 and 1193 kJ/kg BW0.6 /day, respectively. The FHP of pigs averaged 843 kJ/kg BW0.6 /day and was not affected by diet characteristics. A reduction of the respiratory quotients in the fed state as the inclusion level of SBO increased was observed. In conclusion, the NE values of SBO we determined by indirect calorimetry were 33.45 and 34.05 MJ/kg dry matter under two inclusion levels. THP could be largely reduced when SBO is added in the feed, but the THP of SBO included at 5% in a corn-soybean meal diet is not different from the THP of SBO included at 10%.

Advanced glycation end products promote ChREBP expression and cell proliferation in liver cancer cells by increasing reactive oxygen species.

The aim of the study was to elucidate the mechanism by which advanced glycation end products (AGEs) promote cell proliferation in liver cancer cells.We treated liver cancer HepG2 cells with 200 mg/L AGEs or bovine serum albumin (BSA) and assayed for cell viability, cell cycle, and apoptosis. We performed real-time PCR and Western blot analysis for RNA and protein levels of carbohydrate responsive element-binding protein (ChREBP) in AGEs- or BSA-treated HepG2 cells. We analyzed the level of reactive oxygen species (ROS) in HepG2 cells treated with AGEs or BSA.We found that increased S-phase cell percentage and decreased apoptosis contributed to AGEs-induced liver cancer cell proliferation. Real-time PCR and Western blot analysis showed that AGEs stimulated RNA and protein levels of ChREBP, a transcription factor promoting glycolysis and maintaining cell proliferation in liver cancer cells. Intriguingly, the level of ROS was higher in AGEs-treated liver cancer cells. Treating liver cancer cells with antioxidant N-acetyl cystein (NAC) partly blocked AGEs-induced ChREBP expression and cell proliferation.Our results suggest that the AGEs-ROS-ChREBP pathway plays a critical role in promoting ChREBP expression and liver cancer cell proliferation.

Net energy of corn, soybean meal and rapeseed meal in growing pigs.

BACKGROUND: Two experiments were conducted to estimate the net energy (NE) of corn, soybean meal, expeller-pressed rapeseed meal (EP-RSM) and solvent-extracted rapeseed meal (SE-RSM) using indirect calorimetry and to validate the NE of these four ingredients using pig growth performance. METHODS: In Exp.1, 24 barrows (initial BW = 36.4 ± 1.6 kg) were allotted to 1 of 4 diets which included a corn basal diet, a corn-soybean meal basal diet and two rapeseed meal diets containing 20% EP-RSM (9.5% ether extract) or SE-RSM (1.1% ether extract) substituted for corn and soybean meal. The design allowed the calculation of NE values of corn, soybean meal and rapeseed meals according to the difference method. In Exp.2, 175 growing pigs (initial BW = 36.0 ± 5.2 kg) were fed 1 of 5 diets for 28 d, with five pigs per pen and seven replications (pens) per treatment in order to validate the measured energy values. Diets were a corn-soybean meal diet and four diets including 10% or 20% EP-RSM and 10% or 20% SE-RSM. RESULTS: The NE of corn, soybean meal, EP-RSM and SE-RSM were 12.46, 11.34, 11.71 and 8.83 MJ/kg DM, respectively. The NE to ME ratio of corn (78%) was similar to tabular values, however, the NE to ME ratios of soybean meal (70%) and rapeseed meal (76%) were greater than tabular values. The greater NE value in EP-RSM than in SE-RSM is consistent with its higher EE content. Increasing EP-RSM or SE-RSM did not affect the growth performance of pigs and the caloric efficiency of NE was comparable for all diets. CONCLUSIONS: The NE of EP-RSM was similar to soybean meal, and both were greater than SE-RSM. The DE, ME and NE values measured in Exp.1 are confirmed by results of Exp. 2 with comparable caloric efficiencies of DE, ME or NE for all diets.