Changes In Metabolic Phenotype Research Articles

Four cypermethrin isomers induced stereoselective metabolism in H295R cells.

Cypermethrin (CP) is widely used for controlling agricultural and indoor vermin. Previous studies have reported the stereoselective difference of CP in biological activities. However, little is known about their potential mechanisms between metabolic phenotypes and endocrine-disrupting effects. Herein, nuclear magnetic resonance (NMR)-based metabolomics combining metabolite identification and pathway analysis were applied to evaluate the stereoselective metabolic cdisorders induced by CP isomers in human adrenocortical carcinoma cells (H295R) culture medium. Then, gene expression levels related to disturbed metabolic pathways were assessed to verify according to metabolic phenotypes. Metabolomics profiles showed that [(S)-cyano(3-phenoxyphenyl)methyl](1R,3R)-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropane-1-carboxylate [(1R,3R,αS)-CP] induced the most significant changes in metabolic phenotypes than did the other stereoisomers. There are 10 differential metabolites (isoleucine, valine, leucine, ethanol, alanine, acetate, aspartate, arginine, lactate, and glucose) as well as two significantly disturbed pathways, including "pyruvate metabolism" and "alanine, aspartate, and glutamate metabolism," that were confirmed in H295R cells culture medium of (1R,3R,αS)-CP compared with other stereoisomers. Polymerase chain reaction (PCR) array also confirmed the results of metabolomics. Our results can help to understand the potential mechanisms between the isomer selectivity in metabolic phenotypes and endocrine-disrupting effects. Data provided here not only lend authenticity to the cautions issued by the scientists and researchers but also offer a solution for the balance between environment and political regulations.

РОЛЬ МАРКЕРА КОСТНОГО РЕМОДЕЛИРОВАНИЯ ОСТЕОКАЛЬЦИНА В РЕГУЛЯЦИИ ЭНЕРГЕТИЧЕСКОГО ГОМЕОСТАЗА ПРИ САХАРНОМ ДИАБЕТЕ 2 ТИПА

Osteocalcin (OK) is actively involved in the humoral regulation of energy homeostasis. However, the relationship between the level of OK as a modulator of metabolic processes and constitutional and metabolic features in patients with type 2 diabetes mellitus (DM) of a different gender remains not thoroughly studied.The study included 127 patients with type 2 diabetes ≥ 50 years of age. Of these, 70 were postmenopausal women and 57 men.It was found that in the general group of women, the concentration of OK in the blood serum was significantly higher than in men. The observed difference is due to significantly higher levels of OK in women of the older age group (≥ 60 years) in comparison with men. At the same time, a decrease in bone mineral density (BMD) in the femoral neck was observed in subgroups of men and women aged ≥ 60 years and older, while in the younger subgroups of patients, the BMD of lumbar and femoral zones were close to each other.The relationships between OK levels and adipose tissue parameters, evaluated by calculating the morphological and functional index of visceral obesity (IVO), were investigated. An increase in the OK level in the groups of men and women was accompanied by a decrease in the IVO values. The highest degree of insulin resistance was determined in groups of patients with minimal levels of OK and high IVO, and the lowest values were recorded in patients with high levels of OK and low IVO.The decrease of the blood OK level in patients with type 2 diabetes occurs in parallel with an increase in the degree of insulin resistance and dysfunction of visceral adipose tissue. In this case, IVO is a more accurate parameter reflecting the constitutional and metabolic phenotypic changes, compared with the index of the waist circumference. The decrease in BMD in patients with type 2 diabetes is the result of predominantly involutive processes that are noticeable at the age of ≥ 60 years and occur against the background of a decrease in the content of OK with age.

Selenium Regulates Hypothalamic Control of Energy Metabolism in a Sexually Dimorphic Manner

Abstract Objectives The trace element selenium (Se) is known mainly for its antioxidant properties and is critical for proper brain function. The role of Se in regulating energy metabolism, and the sexually dimorphic nature of Se functions, however, are underappreciated, and warrant increased attention. Recent work in our lab has highlighted the importance of Se utilization in hypothalamic regulation of energy metabolism. Dietary Se is incorporated into selenoproteins in the form of the unique amino acid selenocysteine (Sec). The objective of this study was to assess the role of selenoproteins in Agouti-related peptide (Agrp)-positive neurons, an orexigenic sub-population of the hypothalamus. Methods We generated mice with Agrp-Cre-driven deletion of selenocysteine tRNA (Trsp-Agrp KO mice), which is essential for Sec incorporation into selenoproteins, thus ablating selenoprotein synthesis in Agrp-positive neurons. The metabolic phenotype of Trsp-Agrp KO mice challenged with a high-fat diet was characterized via glucose tolerance test (i.p. injection) and the use of analytical chambers to measure food intake and respiratory metabolism. Prior to sacrifice, mice were challenged with leptin (i.p. injection) to assess neuronal leptin responsivity via immunohistochemistry and western blot. Brown adipose tissue (BAT) morphology and thermogenic protein expression were also analyzed. Results Female Trsp-Agrp KO mice displayed resistance to diet-induced obesity, which was accompanied by improved glucose tolerance and elevated energy expenditure levels without changes in food intake. Female Trsp-Agrp KO mice also had greater leptin sensitivity and showed signs of elevated BAT thermogenesis. Male Trsp-Agrp KO mice displayed no changes in metabolic phenotype. Conclusions Loss of selenoproteins in Agrp-positive neurons of the hypothalamus promotes energy expenditure and reduces diet-induced obesity in a sexually dimorphic manner, leading to resistance to a high-fat diet in females. Funding Sources This work was funded by grant support from the National Institute of Diabetes and Digestive and Kidney Diseases (MJB) and Ola HAWAII, a grant from the National Institute on Minority Health and Health Disparities.

Manipulating plasma thyroid hormone levels at hatching alters development of endothermy and ventilation in Pekin duck (Anas platyrhynchos domestica).

At hatching in precocial birds, there are rapid physiological and metabolic phenotypic changes associated with attaining endothermy. During the transition to ex ovo life, thyroid hormone levels naturally increase, peaking at hatching, and then decline. To better understand the role of the natural increase in thyroid hormone at hatching in regulating the developmental trajectory of the Pekin duck's endothermic phenotype, we examined development of O2 consumption (V̇O2 ) and ventilation (frequency, tidal volume and minute ventilation) while inhibiting the developmental increase in thyroid hormones that occurs at hatching via administration of the thyroid-peroxidase inhibitor methimazole (MMI) or accelerating the developmental increase via triiodothyronine (T3) supplementation. Animals were dosed only on day24 of a 28-day incubation period and studied on incubation day25, during external pipping (EP) and 1 day post-hatching (dph). On day25, there was an increase in V̇O2 in the hyperthyroid treatment compared with the other two treatments. During the EP stage, there was a significant effect of thyroid status on V̇O2 ,with hyperthyroid V̇O2 being highest and hypothyroid V̇O2 the lowest. By 1 dph, the supplemented T3 and control animals had similar V̇O2 responses to cooling with comparable thermal neutral zones followed by increased V̇O2 Hypothyroid 1 dph hatchlings had a lower resting V̇O2 that did not increase to the same extent as the supplemented T3 and control animals during cooling. During EP, inhibiting the rise in T3 resulted in embryos with lower ventilation frequency and tidal volume than control and supplemented T3 embryos. At 1 dph, ventilation frequency of all animals increased during cooling, but tidal volume only increased in supplemented T3 and control hatchlings. Our data support the role of the late incubation increase in T3 in regulating the systemic development of endothermic metabolic capacity and associated control of ventilation occurring at hatching of the Pekin duck.

NMR-based metabolomics analysis identifies discriminatory metabolic disturbances in tissue and biofluid samples for progressive prostate cancer

BackgroundProstate cancer (PCa) is one of the most common cancers in men, but its metabolic characteristics during tumor progression are still far from being fully understood. MethodsThe metabolic profiles of matched tissue, serum and urine samples from the same patients were analyzed using a 1H NMR-based metabolomics approach. We identified several important metabolites that significantly altered at different stages of PCa, including benign prostatic hyperplasia (BPH), early PCa (EPC), advanced PCa (APC), metastatic PCa (MPC) and castration-resistant PCa (CRPC). Metabolic correlation networks among tissue, serum and urine samples were examined using Pearson’s correlation. ResultsThe changes in metabolic phenotypes during the progression of PCa were more noticeable in tissue samples when compared with serum and urine samples. Herein we identified a series of important metabolic disturbances, including decreased trends of citrate, creatinine, acetate, leucine, valine, glycine, lysine, histidine, glutamine and choline as well as increased trends of uridine and formate. These metabolites are mainly implicated in energy metabolism, amino acid metabolism, choline and fatty acid metabolism as well as uridine metabolism. We also found that energy metabolism in tumor tissues was positively associated with amino acid metabolism in serum and urine. Additionally, CRPC patients had a peculiar metabolic phenotype, especially decreased amino acid metabolism in serum. ConclusionsThe present study characterizes metabolic disturbances in both tissue and biofluid samples during PCa progression and provides potential diagnostic biomarkers and therapeutic targets for PCa.

SUN-474 Estradiol Protects Against High-Fat Diet-Induced Obesity and Anxiety in Female Mice

Menopause is characterized by a decline in ovarian function and a decrease in estrogens. This decrease in estrogens leads to changes in metabolic phenotype, increasing the risk of weight gain from fat, metabolic syndrome, and type 2 diabetes in postmenopausal women. Menopause is also associated with changes in anxiety and mood. Estradiol (E) exerts similar metabolic effects in female mice as in women. We and others have shown that ovariectomized mice fed a high-fat diet (HFD) become obese, while mice treated with E remain lean. Furthermore, in female mice, HFD increases anxiety, and E acts in specific brain regions to influence anxiety and energy metabolism. However, the mechanisms by which E and diet interact to affect anxiety in females are not well understood. The present study tested the hypothesis that E protects against HFD-induced anxiety-like behavior by altering neural activity in brain regions involved in energy regulation and anxiety. Eight-week-old C57BL6J mice were ovariectomized and implanted with capsules containing either E (50 μg E in 25 μl of 5% ETOH/sesame oil) or vehicle (V). Within each treatment group, mice were fed a standard diet (SD) for 10 days and then maintained on SD or switched to HFD for 26 more days. Mice were weighed and food intake was measured every three days. Starting on day 30, anxiety-like behavior was assessed with three behavior tests: 1) light-dark 2) open-field and 3) elevated plus maze, with an interval of 3 days between each test. To investigate the effects of HFD and E on anxiety-induced neural activity, animals were perfused after the last behavior test. Whole brains were cleared, labeled and imaged for Fos using immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO). The V mice on HFD weighed at least 32% more than all other groups on day 36, while E mice remained lean and had reduced food intake, indicating that E protects against HFD-induced obesity. HFD increased anxiety behavior in the elevated plus maze test as HFD-fed mice spent less time in the open arms (p=0.03). E reduced anxiety-like behavior in mice fed a HFD as measured by the light-dark test. E mice on HFD spent more time in the light compartment than V mice on HFD (p=0.02). E decreased anxiety-like behavior in the open-field test, as E mice entered the center zone more times than V mice (p=0.048). Interestingly, in mice fed a HFD, E decreased Fos expression in the paraventricular hypothalamic nucleus (q<0.1), bed nucleus of the stria terminalis (q<0.05), and the subparafascicular area of the thalamus (q<0.001), suggesting that E protects against HFD-induced anxiety by decreasing neural activity in these specific brain regions. These findings provide insight into the potential mechanisms by which estrogens and diet interact to regulate energy homeostasis and anxiety in postmenopausal women. Work was funded by Otsuka Pharmaceutical Development & Commercialization, Inc.

Pu-erh Tea Regulates Fatty Acid Metabolism in Mice Under High-Fat Diet.

Pu-erh tea has been extensively reported to possess lipid lowering effects but the underlying mechanisms remained unclear. Free fatty acids (FFAs) are generally correlated with the development of obesity, leading to increased risk for type 2 diabetes mellitus and cardiovascular diseases. To investigate whether Pu-erh tea treatment alters FA metabolism, we treated HFD induced obese mice with Pu-erh tea for 22 weeks and analyzed FFA profiles of experimental mice using a UPLC-QTOF-MS platform. Results showed remarkable changes in metabolic phenotypes and FFA compositions in mice treated with or without Pu-erh tea. HFD induced a marked obese phenotype in mice as revealed by significantly increased body weight, liver and adipose tissue weight, lipid levels in serum and liver, and these parameters were markedly reduced by Pu-erh tea treatment. Several FFA or FFA ratios, such as DGLA, palmitoleic acid, and OA/SA ratio, were significantly increased while the levels of SA/PA and AA/DGLA were significantly reduced in HFD-induced obese mice. Interestingly, these differential FFAs or FFA ratios were previous identified as key markers in human obese subjects, and their changes observed in the HFD group were reversed by Pu-erh tea treatment. Moreover, a panel of FFA markers including C20:3 n6/C18:3 n6 and C20:3 n6/C20:2 n6, C18:3 n6/C18:2 n6, C18:3 n3/C18:2 n6 and C24:1 n9/C22:1 n9, which were previously identified as biomarkers in predicting the remission of obesity and diabetes in human subjects who underwent metabolic surgery procedures, were reversed by Pu-erh tea intervention. Pu-erh tea significantly improved glucose homeostasis and insulin tolerance compared to the HFD group. Additionally, Pu-erh tea treatment significantly decreased FFA synthesis genes and increased the expression of genes involved in FFA uptake and β-oxidation including FATP2, FATP5, PPARα, CPT1α, and ACOX-1. These finding confirmed the beneficial effects of Pu-erh tea on regulating lipid and glucose metabolism, and further validated a panel of FFA markers with diagnostic and prognostic value for obesity and diabetes.

High throughput metabolomics-proteomics investigation on metabolic phenotype changes in rats caused by Radix Scrophulariae using ultra-performance liquid chromatography with mass spectrometry.

Radix Scrophulariae, a traditional Chinese herb, is used to treat various diseases, including H2O2-induced apoptosis in cardiomyocytes, HaCaT cells, hyperuricaemia, and depression. This study screened metabolites, proteins and common pathways to better understand both the therapeutic effects and side effects of this herb. Methods: Untargeted metabolomics based on UPLC-TOF-MS, coupled with proteomics based on nano-UPLC-Q-Exactive-MS/MS, were used to investigate the effects of R. Scrophulariae in rats. Fifty-one identified metabolites in urine samples and 76 modulated proteins in liver tissue were potential biomarkers for R. Scrophulariae treatment. The biomarkers and common pathways involved were steroid hormone biosynthesis, drug metabolism-cytochrome p450, drug metabolism-other enzymes, pentose and glucuronate interconversions, and starch and sucrose metabolism. Some biomarkers were beneficial for treating diseases such as cancer, tuberculosis and isovaleric acidaemia, while other biomarkers caused side effects. Metabolomic and proteomic analyses of R. Scrophulariae-treated rats provided valuable information on the biological safety and efficacy of using R. Scrophulariae clinically.

Active Acupoints Differ from Inactive Acupoints in Modulating Key Plasmatic Metabolites of Hypertension: A Targeted Metabolomics Study

The effect of active acupoints versus inactive acupoints in treating hypertension is not well documented. Metabolic phenotypes, depicted by metabolomics analysis, reflect the influence of external exposures, nutrition, and lifestyle on the integrated system of the human body. Therefore, we utilized high-performance liquid chromatography tandem mass spectrometry to compare the targeted metabolic phenotype changes induced by two different acupoint treatments. The clinical outcomes show that active acupoint treatment significantly lowers 24-hour systolic blood pressure but not diastolic blood pressure, as compared with inactive acupoint treatment. Furthermore, distinctive changes are observed between the metabolomics data of the two groups. Multivariate analysis shows that only in the active acupoint treatment group can the follow-up plasma be clearly separated from the baseline plasma. Moreover, the follow-up plasma of these two groups can be clearly separated, indicating two different post-treatment metabolic phenotypes. Three metabolites, sucrose, cellobiose, and hypoxanthine, are shown to be the most important features of active acupoint treatment. This study demonstrates that metabolomic analysis is a potential tool that can be used to efficiently differentiate the effect of active acupoints from inactive acupoints in treating hypertension. Possible mechanisms are the alternation of hypothalamic microinflammation and the restoration of host-gut microbiota interactions induced by acupuncture.

Changes in obese metabolic phenotypes over time and risk of incident chronic kidney disease

To examine the association between metabolically healthy obese (MHO) phenotype and incident chronic kidney disease (CKD) and study whether changes in metabolic phenotypes over time could affect CKD risk. A total of 8589 subjects from the Korean Genome and Epidemiology Study were categorized into four groups based on the presence of obesity and metabolic abnormalities (MA). The primary endpoint was an onset of incident CKD defined as an estimated glomerular filtration rate of ≤ 60 mL/min/1.73 m2 . Multivariable Cox analysis and time-varying Cox analysis were performed to delineate the relationship between obese metabolic phenotypes and incident CKD after adjustment for sociodemographic factors and clinical and laboratory parameters. During a mean follow-up duration of 9.3 years, CKD occurred in 782 (9.1%) participants. In the multivariable Cox model, the hazard ratio (HR) for incident CKD in the MHO, metabolically abnormal non-obese (MANO), and metabolically abnormal obese (MAO) groups was 1.42 (P = 0.002), 1.45 (P < 0.001), and 1.77 (P < 0.001), respectively, compared with the metabolically healthy non-obese (MHNO) group. Time-varying analysis with these four phenotypes as time-varying exposures showed the same results. Furthermore, subjects with persistent MHO through follow-up were at a 2.0-fold increased risk of CKD (P < 0.001). 41.0% of subjects experienced phenotype changes during follow-up. Over the long term, the MHO group had a higher proportion of transition to the MA phenotype and unfavourable metabolic profiles than the MHNO group. Among MHO subjects, those who transitioned to MAO were at a 4.1-fold increased risk of incident CKD than those who regressed to MHNO. In addition, transition to MHO from other groups carried a higher risk of CKD than persistent MHNO. MHO subjects are at increased risk for incident CKD.

Abstract 62: Effect of cancer cell-derived exosome on energy metabolism associated with macrophage activation

Abstract Macrophage is a critical microenvironmental component which can control tumor progression, and dynamic changes in cellular metabolism accompany the polarization. As well as other immune cells, the activation of macrophages can be modulated by cancer cells. Cancer-derived exosome is a newly emerging microenvironmental factor enables cancer cells to communicate with macrophages. In this study, examined were the activation-associated bioenergetic changes and the effect of cancer cell-derived exosome on the changes by a) monitoring the early metabolic kinetics and b) profiling bioenergetic phenotype resulted. In vitro activation of RAW264.7 macrophage cell line by injecting lipopolysaccharide (LPS) and interferon γ (IFNγ) in a Seahorse XF analyzer induced an immediate early increase in glycolytic rate. It also showed a delayed down-regulation of mitochondrial respiration, which resulted in a secondary elevation of glycolytic rate. This secondary glycolytic response was only obtained by LPS-IFNγ co-stimulation and mediated by induced nitric oxide synthase (iNOS) signaling. In contrast to the dynamic changes in metabolic phenotype, the total ATP production rate appeared to be stably maintained according to the XF data normalized by cell number. However, macrophage activation significantly increased glycolysis-dependent portion of ATP production. ATPs were generated almost solely by glycolysis after the delayed mitochondrial down-regulation by iNOS. Co-injection of cancer cell-derived exosomes with LPS slightly but significantly accelerated the immediate early glycolytic response as well as the delayed mitochondrial down-regulation. Furthermore, exosomes induced the immediate early glycolytic response even in the absence of LPS. These results imply that presence of cancer cell-derived exosomes may promote macrophage activation and accelerate pro-inflammatory immune response in a tumor microenvironment. Citation Format: Yoonseok Kam, Brian P. Dranka. Effect of cancer cell-derived exosome on energy metabolism associated with macrophage activation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 62.

Evaluation of the impact of CYP1A2 induction by charbroiled meal on metabolic phenotype

The process of grilling food items often generates polycyclic aromatic hydrocarbons which are established inducers of CYP1A2, a human drug metabolising enzyme, known to activate some procarcinogens. The impact of such induction on CYP1A2 metabolic phenotype has been the subject of some discordant findings. This study, while considering some limitations in previous study designs, evaluated the effect of CYP1A2 induction by the consumption of charbroiled meal on its metabolic phenotype. Caffeine was administered to 17 healthy subjects before, and after, four consecutive days of charbroiled beef ingestion. Blood and spot urine samples were subsequently collected at the 4th and 6th hour post caffeine-administration, respectively, for the assessment of CYP1A2 activity. An additional caffeine administration and sample collection was repeated 48h after the cessation of charbroiled-beef intake. CYP1A2 activity, derived as the log-transformed molar ratios of caffeine and its metabolites, was statistically analysed for changes in metabolic phenotype. Urinary and plasma metrics of CYP1A2 activity had mean reference values of 1.53 and 0.38, respectively, in the study subjects. CYP1A2 metabolic phenotype before and after the ingestion of charbroiled meal was not significantly different. However, urinary and plasma metrics of CYP1A2 activity decreased by about 19% (1.53 vs 1.24) and 65% (0.38 vs 0.14), respectively, 48h after the cessation of charbroiled meal ingestion. The induction of CYP1A2 by the consumption of charbroiled meals may not portend increased rate of CYP1A2-activation of procarcinogens in humans. However, a potentially significant CYP1A2 inhibition which might result in increased-exposure for drugs predominantly metabolised by this enzyme is likely.

Changes in metabolic phenotype and cellular ATP production during CD4+ T cell activation.

Abstract Activation of CD4+ T cells is followed by rapid proliferation and differentiation into specific subsets (Treg or Teff). These transitions are accompanied by tight regulated changes in energetic demand and cellular metabolic reprogramming. Using Agilent Seahorse extracellular flux analysis, we have developed a cell-based assay for simultaneous measurement of the two-main cellular ATP-producing pathways, i.e. glycolysis and oxidative phosphorylation. The assay allows for quantification of real time changes in total ATP production rate, and the fractional contribution of the individual pathways to support bioenergetic demands. We found that naïve CD4+ T cells obtain most of its ATP from mitochondrial oxidative phosphorylation. After activation with anti CD3/CD28 conjugated beads, total ATP production rate significantly increases, and that rise is sustained by an increase in glycolytic ATP production, but also in mitochondrial ATP production. When glucose is replaced by galactose in the assay medium, T cell activation is inhibited despite partial compensation of total ATP production through mitochondrial respiration. Time-course of bioenergetic phenotype during T-cell expansion shows a deep increase in ATP production rate up to day 6–7 with glycolysis contributing up to 70% of total ATP production. When glucose is replaced by galactose 72 hr after activation, total ATP production rate was fully compensated by mitochondrial ATP production but still the inhibition of glycolytic activity by galactose reduces cell proliferation. The results obtained highlight the role of glycolytic activity beyond energetic supply and its importance for biosynthetic and redox requirements or cell activation and proliferation.

Impact of Exercise and Aging on Rat Urine and Blood Metabolome. An LC-MS Based Metabolomics Longitudinal Study.

Aging is an inevitable condition leading to health deterioration and death. Regular physical exercise can moderate the metabolic phenotype changes of aging. However, only a small number of metabolomics-based studies provide data on the effect of exercise along with aging. Here, urine and whole blood samples from Wistar rats were analyzed in a longitudinal study to explore metabolic alterations due to exercise and aging. The study comprised three different programs of exercises, including a life-long protocol which started at the age of 5 months and ended at the age of 21 months. An acute exercise session was also evaluated. Urine and whole blood samples were collected at different time points and were analyzed by LC-MS/MS (Liquid Chromatography–tandem Mass Spectrometry). Based on their metabolic profiles, samples from trained and sedentary rats were differentiated. The impact on the metabolome was found to depend on the length of exercise period with acute exercise also showing significant changes. Metabolic alterations due to aging were equally pronounced in sedentary and trained rats in both urine and blood analyzed samples.

Metabolomic analysis of human oral cancer cells with adenylate kinase 2 or phosphorylate glycerol kinase 1 inhibition.

The purpose of this study was to use liquid chromatography-mass spectrometry (LC-MS) with XCMS for a quantitative metabolomic analysis of UM1 and UM2 oral cancer cells after knockdown of metabolic enzyme adenylate kinase 2 (AK2) or phosphorylate glycerol kinase 1 (PGK1). UM1 and UM2 cells were initially transfected with AK2 siRNA, PGK1 siRNA or scrambled control siRNA, and then analyzed with LC-MS for metabolic profiles. XCMS analysis of the untargeted metabolomics data revealed a total of 3200-4700 metabolite features from the transfected UM1 or UM2 cancer cells and 369-585 significantly changed metabolites due to AK2 or PGK1 suppression. In addition, cluster analysis showed that a common group of metabolites were altered by AK2 knockdown or by PGK1 knockdown between the UM1 and UM2 cells. However, the set of significantly changed metabolites due to AK2 knockdown was found to be distinct from those significantly changed by PGK1 knockdown. Our study has demonstrated that LC-MS with XCMS is an efficient tool for metabolomic analysis of oral cancer cells, and knockdown of different genes results in distinct changes in metabolic phenotypes in oral cancer cells.

PUB059 Role of Alveolar Macrophages' Molecular Antennas in the Pathogenesis of Lung Cancer

The internal sources of alpha-emitting radionuclides originated from tobacco smoke or inhaled contaminated aerosol, induce the low-slow dose effects in pulmonary tissue. The health effects may not appear for years. The metabolic, ionic and enzyme properties of alveolar macrophages (AM), represent in situ biochemical, immunological and signaling processes. Apoptotic regulation is important for tissue remodeling. The study was performed in order to elucidate some of the mechanisms included in the pathogenesis of lung cancer. Bronchoalveolar lavage (BAL) was performed in a group of smokers whose average smoking exposure (pack-years) values were < 2, and patients with non-small cell lung cancer (NSCLC) > 40, as well as in the control group of nonsmokers. Differential cell counting and cytochemical analysis of the BAL specimens for intracellular iron content, lipids, glycogen, nonspecific esterases, and acid phosphatase (ACP) were performed by light microscopy. Apoptosis detection was performed by using the TUNEL in situ cytochemical method. Iron content in AM is gradually increasing in smokers, as well as in patients with NSCLC, (p<0.05) in comparison with non-smokers. There were no significant differences for ACP, nonspecific esterases, glycogen and lipid content in AM between the investigated groups. Apoptosis is gradually increasing in smokers and NSCLC group in comparison with nonsmokers. Apoptotic clearance of free apoptotic bodies by AM phagocytosis is increased in smokers in comparison with nonsmokers, but in the NSCLC group, AC is significantly decreased both, in comparison with nonsmokers, as well as control smokers (p<0.05). AM possess iron centers in their enzymes. The regulation of gene expression for ACP depends on iron. The binuclear iron center of ACP is included in the generation of reactive oxygen species. During the prolonged tissue injury, AM gather iron which can itself contribute to the process of cell damage, as an ionic part of molecular antennas (biomolecules, including enzymes with metal ion surrounded by biopolymer chains). Ferroptosis could be an iron-dependent form of non-apoptotic death of AM, without cell energy depletion. Alpha-emitting internal sources of radiation may trigger random and irregular tissue damage and repair. Oxygen diffusion limit increases for the population of proliferating cells. Their metabolic phenotype changes from glycolytic to lipogenic, with the further transformation of cells to the initiation of neoplastic lesion in the prolonged course. The changed biological properties of AM could be of importance as a utility for biological dosimetry purposes.

Gut microbiota after Roux-en-Y gastric bypass and sleeve gastrectomy in a diabetic rat model: Increased diversity and associations of discriminant genera with metabolic changes.

Recent work with gut microbiota after bariatric surgery is limited, and the results have not been in agreement. Given the role of the gut microbiota in regulating host metabolism, we explored the effect of Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy (SG) on the modifications of gut microbiota with regard to the potential influence of food intake and/or weight loss and examined their links with host metabolism. Zucker diabetic fatty rats were divided into the following groups: RYGB; sham-operated with pair-fed as RYGB; sham-operated fed ad libitum; and SG. The metabolic effects and gut microbiota profile were analyzed 10weeks postoperatively. Associations between discriminating genera and metabolic markers after RYGB were explored. The 2 procedures induced similar glucose improvement and increased flora diversity after 10weeks compared with sham-operated groups. RYGB induced a marked higher relative abundance of Proteobacteria/Gammaproteobacteria and Betaproteobacteria and increased emergence of Fusobacteria and Clostridium, whereas SG resulted in more abundant Actinobacteria compared with other groups. Most of the 12 discriminant genera correlated with changes in metabolic phenotype, but only 28.6% of these correlations were independent of weight, and 4 discriminant genera still negatively correlated with serum insulin level independent of food intake and weight loss after RYGB. These data demonstrate that RYGB and SG surgery produced similar diversity but different microbiota compositions changes in Zucker diabetic fatty rats. These findings stimulate deeper explorations of functions of the discriminate microbiota and the mechanisms linking postsurgical modulation of gut microbiota and improvements in insulin resistance.

Transgenerational changes of metabolic phenotypes in two selectively bred mouse colonies for different susceptibilities to diet-induced glucose intolerance

We recently established 2 mouse lines with different susceptibilities (prone and resistant) to high-fat diet (HFD)-induced glucose intolerance by selective breeding (designated selectively bred diet-induced glucose intolerance-prone [SDG-P] and -resistant [SDG-R], respectively). In the present study, we analyzed transgenerational changes in metabolic phenotypes in these 2 mouse colonies to explore how the distinct phenotypes have emerged through the repetitive selection. Using C57BL/6, C3H, and AKR as background strains, mice showing inferior and superior glucose tolerance after HFD feeding were selected and bred repetitively over 20 generations to produce SDG-P and SDG-R, respectively. In addition to the blood glucose levels, HFD intake and body weight were also measured over the selective breeding period. As the generations proceeded, SDG-P mice became more susceptible to HFD-induced glucose intolerance and body weight gain, whereas SDG-R mice had gradually reduced HFD intake. The differences in fasting and post-glucose challenge blood glucose levels, body weight, and HFD intake became more evident between the 2 colonies through the selective breeding, mainly due to the HFD-induced glucose metabolism impairment and body weight gain in SDG-P mice and the reduction of HFD intake in SDG-R mice. These transgenerational changes in the metabolic phenotypes suggest that the genetic loci associated with the quantitative traits have been selectively enriched in SDG-P and SDG-R.

Urinary Metabolic Profiling of ‘MARS-500 Project’ with Nuclear Magnetic Resonance Spectroscopy

Human sustainable space exploration could go further with the development of strategies and preparatory activities. Enormous scientific work has been done during the 21st century. MARS-500 project is one of unique preparatory activities, and was finished within 520 day isolation on the Earth. The full 520 day MARS mission simulation provided an opportunity to conduct a long-term, isolated experiment to observe metabolic changes due to people differences and its stability to duration variation. Six volunteers were selected from Russia, France, Italy, and China, and their urine specimens were collected in six periods. Then the subtle metabolic variations were investigated by nuclear magnetic resonance (NMR) at six different periods. We found that the metabotypes of six crews drifted out from the first period to the farthest third period, then reverted to the nearest sixth period in metabolic space, suggesting a property of resilience for metabolic phenotype. Furthermore, a set of 20 biomarkers, discriminating great changes in metabolic phenotypes well, were associated with emotion, muscle turnover, circadian rhythms, energy expenditure, lipid metabolism, oxidative activity, cognitive performance, and gut-microbial activities. In deeps, to analysis the data from MARS-500 project with two-way ANOVA, an interaction between factor of people differences and long-term isolation showed that the six crews each had a specific response to 520 days’ isolation. In this picture, accurate healthy management during future human space mission should aim at promoting his or her ability to revert to his or her stable metabotype under a defined and isolated environment.

Changes in metabolic phenotypes of Plasmodium falciparum in vitro cultures during gametocyte development.

BackgroundGametocytes are the Plasmodium life stage that is solely responsible for malaria transmission. Despite their important role in perpetuating malaria, gametocyte differentiation and development is poorly understood.MethodsTo shed light on the biochemical changes that occur during asexual and gametocyte development, metabolic characterization of media from in vitro intra-erythrocytic Plasmodium falciparum cultures was performed throughout gametocyte development by applying 1H nuclear magnetic spectroscopy, and using sham erythrocyte cultures as controls. Spectral differences between parasite and sham cultures were assessed via principal component analyses and partial-least squares analyses, and univariate statistical methods.ResultsClear parasite-associated changes in metabolism were observed throughout the culture period, revealing differences between asexual parasites and gametocyte stages. With culture progression and development of gametocytes, parasitic release of the glycolytic end products lactate, pyruvate, alanine, and glycerol, were found to be dramatically reduced whilst acetate release was greatly increased. Also, uptake of lipid moieties CH2, CH3, and CH = CH-CH2-CH2 increased throughout gametocyte development, peaking with maturity.ConclusionsThis study uniquely presents an initial characterization of the metabolic exchange between parasite and culture medium during in vitro P. falciparum gametocyte culture. Results suggest that energy metabolism and lipid utilization between the asexual stages and gametocytes is different. This study provides new insights for gametocyte-specific nutritional requirements to aid future optimization and standardization of in vitro gametocyte cultivation, and highlights areas of novel gametocyte cell biology that deserve to be studied in greater detail and may yield new targets for transmission-blocking drugs.Electronic supplementary materialThe online version of this article (doi:10.1186/1475-2875-13-468) contains supplementary material, which is available to authorized users.