Concentrations Of Glycerophosphocholine Research Articles

Perfluorohexanesulfonic Acid (PFHxS) Impairs Lipid Homeostasis in Zebrafish Larvae through Activation of PPARα.

Perfluorohexanesulfonic acid (PFHxS), an emerging short-chain per- and polyfluoroalkyl substance, has been frequently detected in aquatic environments. Adverse outcome pathway studies have shown that perfluorinated compounds impair lipid homeostasis through peroxisome proliferator activated receptors (PPARs). However, many of these studies were performed at high concentrations and may thus be a result of overt toxicity. To better characterize the molecular and key events of PFHxS to biota, early life-stage zebrafish (Danio rerio) were exposed to concentrations detected in the environment (0.01, 0.1, 1, and 10 μg/L). Lipidomic and transcriptomic evaluations were integrated to predict potential molecular targets. PFHxS significantly impaired lipid homeostasis by the dysregulation of glycerophospholipids, fatty acyls, glycerolipids, sphingolipids, prenol lipids, and sterol lipids. Informatic analyses of the lipidome and transcriptome indicated alterations of the PPAR signaling pathway, with downstream changes to retinol, linoleic acid, and glycerophospholipid metabolism. To assess the role of PPARs, potential binding of PFHxS to PPARs was predicted and animals were coexposed to a PPAR antagonist (GW6471). Molecular simulation indicated PFHxS had a 27.1% better binding affinity than oleic acid, an endogenous agonist of PPARα. Antagonist coexposures rescued impaired glycerophosphocholine concentrations altered by PFHxS. These data indicate PPARα activation may be an important molecular initiating event for PFHxS.

Autologous hematopoietic stem cell transplantation significantly alters circulating ceramides in peripheral blood of relapsing-remitting multiple sclerosis patients

BackgroundThe common inflammatory disease multiple sclerosis (MS) is a disease of the central nervous system. For more than 25 years autologous hematopoietic stem cell transplantation (AHSCT) has been used to treat MS. It has been shown to be highly effective in suppressing inflammatory activity in relapsing-remitting MS (RRMS) patients. This treatment is thought to lead to an immune system reset, inducing a new, more tolerant system; however, the precise mechanism behind the treatment effect in MS patients is unknown. In this study, the effect of AHSCT on the metabolome and lipidome in peripheral blood from RRMS patients was investigated.MethodsPeripheral blood samples were collected from 16 patients with RRMS at ten-time points over the five months course of AHSCT and 16 MS patients not treated with AHSCT. Metabolomics and lipidomics analysis were performed using liquid-chromatography high-resolution mass spectrometry. Mixed linear models, differential expression analysis, and cluster analysis were used to identify differentially expressed features and groups of features that could be of interest. Finally, in-house and in-silico libraries were used for feature identification, and enrichment analysis was performed.ResultsDifferential expression analysis found 657 features in the lipidomics dataset and 34 in the metabolomics dataset to be differentially expressed throughout AHSCT. The administration of cyclophosphamide during mobilization and conditioning was associated with decreased concentrations in glycerophosphoinositol species. Thymoglobuline administration was associated with an increase in ceramide and glycerophosphoethanolamine species. After the conditioning regimen, a decrease in glycerosphingoidlipids concentration was observed, and following hematopoietic stem cell reinfusion glycerophosphocholine concentrations decreased for a short period of time. Ceramide concentrations were strongly associated with leukocyte levels during the procedure. The ceramides Cer(d19:1/14:0) and Cer(d20:1/12:0) were found to be increased (P < .05) in concentration at the three-month follow-up compared to baseline. C16 ceramide, Cer(D18:2/16:0), and CerPE(d16:2(4E,6E)/22:0) were found to be significantly increased in concentration after AHSCT compared to prior to treatment as well as compared to newly diagnosed RRMS patients.ConclusionAHSCT had a larger impact on the lipids in peripheral blood compared to metabolites. The variation in lipid concentration reflects the transient changes in the peripheral blood milieu during the treatment, rather than the changes in the immune system that are assumed to be the cause of clinical improvement within RRMS patients treated with AHSCT. Ceramide concentrations were affected by AHSCT and associated with leukocyte counts and were altered three months after treatment, suggesting a long-lasting effect.

Potential role of skeletal muscle glycerophosphocholine in response to altered fluid balance in humans: an in vivo nuclear magnetic resonance study.

Many cells adapt to hyperosmolal conditions by upregulation of organic osmolytes to maintain cell function and integrity. Glycerophosphocholine (GPC), a recognized osmolyte in renal medullary cells, is the major phosphodiester (PDE) in human skeletal muscle, wherefore we hypothesized muscular GPC to be associated with surrogate parameters of fluid status and osmolality in healthy humans. The objective of this study was to investigate the relationship of muscular GPC with surrogate parameters of body fluid status and osmolality. We analyzed data of 30 healthy volunteers who underwent noninvasive 31P-magnetic resonance spectroscopy of either calf (n = 17) or thigh (n = 13) muscle. Therefore, we conducted correlation analyses between phosphor metabolites, and blood values depicting body fluid status and osmolality. Relevant parameters were further implemented in a multivariable regression model to evaluate if GPC concentrations can depict variations in fluid and electrolyte balance. Uric acid (0.437, P = 0.018) and urea (0.387, P = 0.035) were significantly correlated with GPC, which in case of uric acid was independent of sex. Considering sex, following multivariable regression reported GPC as suitable parameter to predict uric acid (R2 = 0.462, adjusted R2 = 0.421; P < 0.001). Our data indicate a connection between muscular GPC concentrations and uric acid, which is a marker of body fluid status, in healthy human subjects, suggesting that skeletal muscle might regulate GPC content in adaptation to changes in fluid status.NEW & NOTEWORTHY Using in vivo magnetic resonance spectroscopy, our study is the first one indicating fluid balance-dependent properties of glycerophosphocholine concentrations in human skeletal muscle. In vivo examination of GPC as organic osmolyte in human skeletal muscle marks a novel approach, which might give further insight on how water and electrolyte balance affect muscle tissue. Beside this main finding, glycerophosphocholine of both calf and thigh muscle correlated remarkably with blood laboratory parameters of lipid metabolism in our study population.

Integrating omics to characterize eco-physiological adaptations: How moose diet and metabolism differ across biogeographic zones.

With accelerated land conversion and global heating at northern latitudes, it becomes crucial to understand, how life histories of animals in extreme environments adapt to these changes. Animals may either adapt by adjusting foraging behavior or through physiological responses, including adjusting their energy metabolism or both. Until now, it has been difficult to study such adaptations in free‐ranging animals due to methodological constraints that prevent extensive spatiotemporal coverage of ecological and physiological data.Through a novel approach of combining DNA‐metabarcoding and nuclear magnetic resonance (NMR)‐based metabolomics, we aim to elucidate the links between diets and metabolism in Scandinavian moose Alces alces over three biogeographic zones using a unique dataset of 265 marked individuals.Based on 17 diet items, we identified four different classes of diet types that match browse species availability in respective ecoregions in northern Sweden. Individuals in the boreal zone consumed predominantly pine and had the least diverse diets, while individuals with highest diet diversity occurred in the coastal areas. Males exhibited lower average diet diversity than females.We identified several molecular markers indicating metabolic constraints linked to diet constraints in terms of food availability during winter. While animals consuming pine had higher lipid, phospocholine, and glycerophosphocholine concentrations in their serum than other diet types, birch‐ and willow/aspen‐rich diets exhibit elevated concentrations of several amino acids. The individuals with highest diet diversity had increased levels of ketone bodies, indicating extensive periods of starvation for these individuals.Our results show how the adaptive capacity of moose at the eco‐physiological level varies over a large eco‐geographic scale and how it responds to land use pressures. In light of extensive ongoing climate and land use changes, these findings pave the way for future scenario building for animal adaptive capacity.

Abstract PS1-38: Role of MR spectroscopy in evaluation of axilla in breast cancer

Abstract The role of surgery for women with breast cancer presenting with clinically negative axilla is being questioned. Trials are underway to evaluate axillary ultrasound (AUS) for non-invasive evaluation of axilla instead of sentinel node biopsy. However, AUS has a sensitivity of 50-59%. Thus, there is a need for non-invasive test to complement AUS. MR Spectoroscopy (MRS) identifies metabolites, viz. glycerophosphocholine (GPC), phosphocholine (PC), threonine (Thr), lactate (Lac), Choline (Cho) etc that are associated with presence of metastasis in axillary lymph nodes (LN) . Biochemical changes in tumor infiltrated LNs occur earlier than morphological changes and MRS can detect these chemical changes, it can be a useful investigation in evaluation of axilla. Aim: To evaluate the role of MR Spectroscopy (MRS) in identifying biochemical changes in axillary lymph nodes in breast cancer and correlate the findings with Histopathology (HP) to predict presence of metastasis.Methods: Axillary lymph nodes were obtained from 59 patients with early breast cancer with clinically negative axilla who underwent surgery. Each LN was bisected into two equal halves. One half of LN was snap frozen in liquid nitrogen (-196°C) and then stored at −80°C until perchloric acid extraction was carried out. Other half of LN was subjected to HP evaluation with standard Eosin and Hematoxylin staining . METHOD OF MR Spectroscopy: Water-soluble metabolites from the tissue samples were extracted using PCA extraction procedure (Seenu et al Magn Reson Imaging 2005 doi: 10.1016/j.mri.2005.10.004). Proton (1H) NMR spectroscopy of specimen was performed. Concentrations of GPC, PC, Thr, Lac, Cho and other metabolites were determined by comparing the integrated intensity of isolated resonances of the compounds of interest with that of the TSP signal, correcting for the number of contributing protons and with the tissue weight . Intensity ratio for metabolites, GPC, PC and Thr (GPC+PC/Thr) was also determined.Statistical Analyses: Levels of concentrations of various metabolites were compared between involved and non-involved lymph nodes using Wilcoxon rank sum test. Diagnostic indices of MRS were assessed in terms of sensitivity, specificity, positive predictive value, negative predictive value, likelihood ratios and overall accuracy taking final histopathology as ‘gold standard’ with 95% confidence intervals.Results: Mean concentrations of GPC, PC and Thr were significantly increased in involved as compared to non-involved node (table 1). Cho and Lac were not significantly increased. A cut-off value of 0.80 for the GPC-PC/Thr ratio was chosen to obtain a maximum accuracy of 89% based on previous study. MRS accurately predicted metastasis in 21 of the 24 patients who had LN metastasis on HP. Out of 35 patients with no LN metastasis, MRS correlated accurately with HP in 30 of them. Sensitivity, specificity, PPV, NPV and overall accuracy for MRS in detecting LN metastasis were 87.5%; 88%; 80.7%; 90.9%; and 86.4% respectively. Likelihood ratio for positive test and negative test for MRS to detect LN metastasis were 6.1 and 0.14 respectively.Conclusion: MRS can accurately predict presence of metastasis in axillary lymph nodes in vitro. In vivo studies are essential to corroborate these findings. Mean concentrations of metabolites in axillary lymph nodesInvolvedNon involvedP valueGPC0.5923(SD=0.5726)0.3648(SD=0.6774)0.02PC0.7347(SD=0.9466)0.5286(SD=1.1111)0.03Cho0.6930(SD=0.6756)0.8435(SD=0.5030)0.21Thr2.9656(SD=2.2397)1.1674(SD=1.2274)0.02Lac4.5515(SD=2.3271)3.2907(SD=1.3296)0.31 Citation Format: Srineil Vuthaluru, Anurag Srivastava. Role of MR spectroscopy in evaluation of axilla in breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS1-38.

Improved Spatial Resolution of Metabolites in Tissue Biopsies Using High-Resolution Magic-Angle-Spinning Slice Localization NMR Spectroscopy.

High-resolution magic-angle-spinning 1H NMR spectroscopy (HR-MAS NMR) is a well-established technique for assessing the biochemical composition of intact tissue samples. In this study, we utilized a method based on HR-MAS NMR spectroscopy with slice localization (SLS) to achieve spatial resolution of metabolites. The obtained 7 slice spectra from each of the model samples (i.e., chicken thigh muscle with skin and murine renal biopsy including medulla (M) and cortex (C)) showed distinct metabolite compositions. Furthermore, we analyzed previously acquired 1H HR-MAS NMR spectra of separated cortex and medulla samples using multivariate statistical methods. Concentrations of glycerophosphocholine (GPC) were found to be significantly higher in the renal medulla compared to the cortex. Using GPC as a biomarker, we identified the tissue slices that were predominantly the cortex or medulla. This study demonstrates that HR-MAS SLS combined with multivariate statistics has the potential for identifying tissue heterogeneity and detailed biochemical characterization of complex tissue samples.

Variability of Water-Soluble Forms of Choline Concentrations in Human Milk during Storage, after Pasteurization, and among Women.

Choline is critical for infant development and mother’s milk is the sole source of choline for fully breastfed infants until six months of age. Human milk choline consists to 85% of water-soluble forms of choline including free choline (FC), phosphocholine (PhosC), and glycerophosphocholine (GPC). Donor milk requires safe handling procedures such as cold storage and pasteurization. However, the stability of water-soluble forms of choline during these processes is not known. The objectives of this research were to determine the effect of storage and pasteurization on milk choline concentration, and the diurnal intra- and inter-individual variability of water-soluble choline forms. Milk samples were collected from healthy women who were fully breastfeeding a full-term, singleton infant <6 months. Milk total water-soluble forms of choline, PhosC, and GPC concentrations did not change during storage at room temperature for up to 4 h. Individual and total water-soluble forms of choline concentrations did not change after storage for 24 h in the refrigerator or for up to one week in the household freezer. Holder pasteurization decreased PhosC and GPC, and thereby total water-soluble choline form concentrations by <5%. We did not observe diurnal variations in PhosC and total water-soluble forms of choline concentrations, but significant differences in FC and GPC concentrations across five sampling time points throughout one day. In conclusion, these outcomes contribute new knowledge for the derivation of evidence-informed guidelines for the handling and storage of expressed human milk as well as the development of optimized milk collection and storage protocols for research studies.

Skeletal muscle metabolism in rats with low and high intrinsic aerobic capacity: Effect of aging and exercise training.

PurposeExercise training increases aerobic capacity and is beneficial for health, whereas low aerobic exercise capacity is a strong independent predictor of cardiovascular disease and premature death. The purpose of the present study was to determine the metabolic profiles in a rat model of inborn low versus high capacity runners (LCR/HCR) and to determine the effect of inborn aerobic capacity, aging, and exercise training on skeletal muscle metabolic profile.MethodsLCR/HCR rats were randomized to high intensity low volume interval treadmill training twice a week or sedentary control for 3 or 11 months before they were sacrificed, at 9 and 18 months of age, respectively. Magnetic resonance spectra were acquired from soleus muscle extracts, and partial least square discriminative analysis was used to determine the differences in metabolic profile.ResultsSedentary HCR rats had 54% and 30% higher VO2max compared to sedentary LCR rats at 9 months and 18 months, respectively. In HCR, exercise increased running speed significantly, and VO2max was higher at age of 9 months, compared to sedentary counterparts. In LCR, changes were small and did not reach the level of significance. The metabolic profile was significantly different in the LCR sedentary group compared to the HCR sedentary group at the age of 9 and 18 months, with higher glutamine and glutamate levels (9 months) and lower lactate level (18 months) in HCR. Irrespective of fitness level, aging was associated with increased soleus muscle concentrations of glycerophosphocholine and glucose. Interval training did not influence metabolic profiles in LCR or HCR rats at any age.ConclusionDifferences in inborn aerobic capacity gave the most marked contrasts in metabolic profile, there were also some changes with ageing. Low volume high intensity interval training twice a week had no detectable effect on metabolic profile.

Abstract 2120: Distinct molecular effects of chemotherapeutic agents on choline phospholipid metabolism of triple-negative breast cancer cells

Abstract The magnetic resonance spectroscopy (MRS)-detected total choline (tCho) signal is a promising non-invasive surrogate marker able to predict chemotherapy response in breast cancer patients early on. However, the molecular mechanisms by which common chemotherapeutic drugs affect the tCho signal, which consists of glycerophosphocholine (GPC), phosphocholine (PC), and free choline (Cho), are mostly unknown. Here we have employed some widely used cancer chemotherapeutic drugs such as doxorubicin, paclitaxel, and vinorelbine to treat triple-negative human MDA-MB-231 breast cancer cells to elucidate their molecular effects on choline phospholipid metabolism. High-resolution (HR) 1H MRS of water-soluble cell extracts was employed to detect changes in cellular choline metabolite profiles, and quantitative RT-PCR (qRT-PCR) to assess the corresponding changes in the expression levels of choline-metabolizing enzymes. After 48 hours of doxorubicin treatment, the GPC levels in MDA-MB-231 cells significantly increased, while PC level decreased, and the tCho concentration remained unchanged. Vinorelbine treatment displayed a comparable effect to doxorubicin, but the GPC increase and PC decrease occurred to a lesser extent. Paclitaxel treatment caused an increased GPC level along with an unaltered PC level, leading to a slightly increased tCho level. In all these cases, the PC/GPC ratio decreased from 3.5 in the control to about 2.0 in paclitaxel- or vinorelbine-treated samples, to about 1.0 in doxorubicin-treated samples. qRT-PCR-detected mRNA expression levels showed that GDPD6 and Chkα were downregulated by doxorubicin. GDPD5, GDPD6, and Chkα genes displayed a reduced mRNA expression level following vinorelbine treatment, while no significant change in these three genes was detected after paclitaxel treatment. For comparison, the known GDPD6 inhibitor dipyridamole significantly increased cellular GPC levels, but, as expected, did not affect GDPD6 mRNA expression levels as it acts as an enzyme inhibitor. Our study demonstrates that choline containing metabolites change differently depending on the type of drug used for breast cancer treatment. However, all tested chemotherapeutic treatments resulted in metabolic alteration away from the ‘cholinic phenotype’, which was caused in some cases by decreases in expression of Chkα, GDPD6, and GDPD5 following drug treatment. The choline metabolite concentrations of GPC and/or PC and/or the PC/GPC ratio may serve as non-invasive surrogate makers of therapeutic response in triple-negative breast cancer patients undergoing chemotherapy. Note: This abstract was not presented at the meeting. Citation Format: Menglin Cheng, Zaver M. Bhujwalla, Kristine Glunde. Distinct molecular effects of chemotherapeutic agents on choline phospholipid metabolism of triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2120. doi:10.1158/1538-7445.AM2017-2120

IMPACT ОF CHRONIC INHALATION OF LOW DOSES OF ALIPHATIC HYDROCARBONS C6-C10 ON METABOLIC PROFILES OF RATS BRAIN AND LIVER

For the first time, the chronic inhalation effect of aliphatic hydrocarbons (HC) in low concentrations on metabolic profiles of low molecular weight organic compounds including free (FFA) and esterified (EFA) fatty acids in rats brain and liver was studied. Gas and liquid chromatography-mass spectrometry was used as analytical methods. The list of hydrocarbons includes hexane, heptane, octane, nonane and decane in various ratios. Changes in profiles of low-molecular compounds in the brain were detected under the action of a mixture of HCs in a concentration of 160 ± 20.5 mg/m3 for 90 days, while the action of the HCs mixture at lower concentrations of 31.4 ± 5.6 mg/m3 and 5.2 ± 1.08 mg/m3 only leads to minor changes in profiles. A significant increase in lysophospholipids concentrations, mainly of lysophosphatidylcholine, in the extract from the brain and blood plasma was revealed. The increase in the content of lysophospholipids in brain tissues can be considered as a consequence of the destruction of cell membranes, specific to neurodegenerative processes. At the same time, an increase in the lysophosphatidylcholine concentration in the brain is accompanied by a statistically significant decrease in the concentration of glycerol-3-phosphate and is not followed by changes in the concentrations of glycerophosphocholine, glycerophosphoethanolamine, choline, acetylcholine, FFA, or lysophosphatidic acid. It was shown that concentrations of lysophosphatidylcholine and lysophosphatidylethanamine, referred to the concentrations of glycerophosphocholine and glycerophosphoethanolamine in blood plasma, may serve as potential markers of the effect of HC on the brain.

Role of NMR metabonomics in Celiac Disease (CeD)

Metabonomics study provides a comprehensive metabolic profile of biological samples using techniques like mass spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. The metabolites identified are later analysed using mul- tivariate statistical methods. Metabonomics has the potential to provide putative biomarker/s for disease diagnosis and for monitoring the disease progression and can be used in patient management. Recently, a few metabonomics studies have been reported on blood sera, urine and intestinal mucosal biopsies of celiac disease (CeD) patients using proton NMR. Significantly decreased levels of amino acids, methylamine, lactate, lipids, pyruvate, creatinine, choline and glycoprotein and increased levels of glucose and β-hydroxybutyrate have been reported in blood sera of CeD patients. In intestinal mucosal biopsies of CeD patients, a higher concentration of isoleucine, leucine, aspartate, succinate and pyruvate and lower concentration of glycerophosphocholine was seen as compared to controls. These studies indicates that the metabonomics study of CeD using in-vitro NMR spectroscopy helps in the determination of metabolic signature/s of the disease. It also provides an insight into the biochemistry of the disease and also helps in the identification of metabolites that could serve as putative biomarker/s for the diagnosis of CeD. This review focuses on the application of NMR based metabonomics in CeD and highlights the potential of NMR based metabonomics in the identification of biomarker/s for diagnosis and prognosis.

Relationships among Different Water-Soluble Choline Compounds Differ between Human Preterm and Donor Milk.

Choline is essential for infant development. Human milk choline is predominately present in three water-soluble choline (WSC) forms: free choline (FC), phosphocholine (PhosC), and glycerophosphocholine (GPC). It is unclear whether mother’s own preterm milk and pooled donor milk differ in WSC composition and whether WSC compounds are interrelated. Mother’s own preterm milk (n = 75) and donor milk (n = 30) samples from the neonatal intensive care unit, BC Women’s Hospital were analyzed for WSC composition using liquid chromatography tandem mass spectrometry (LC-MS/MS). Associations between different WSC compounds were determined using Pearson’s correlations, followed by Fischer r-to-z transformation. Total WSC concentration and concentrations of FC, PhosC, and GPC did not significantly differ between mother’s own milk and donor milk. FC was negatively associated with PhosC and GPC in mother’s own milk (r = −0.27, p = 0.02; r = −0.34, p = 0.003, respectively), but not in donor milk (r = 0.26, p = 0.181 r = 0.37, p = 0.062, respectively). The difference in these associations between the two milk groups were statistically significant (p = 0.03 for the association between PhosC and FC; and p = 0.003 for the association between FC and GPC). PhosC and GPC were positively associated in mother’s own milk (r = 0.32, p = 0.036) but not donor milk (r = 0.36, p = 0.062), although the difference in correlation was not statistically significant. The metabolic and clinical implications of these associations on the preterm infant need to be further elucidated.

Proton and phosphorus magnetic resonance spectroscopy of the healthy human breast at 7T.

In vivo water‐ and fat‐suppressed 1H magnetic resonance spectroscopy (MRS) and 31P magnetic resonance adiabatic multi‐echo spectroscopic imaging were performed at 7 T in duplicate in healthy fibroglandular breast tissue of a group of eight volunteers. The transverse relaxation times of 31P metabolites were determined, and the reproducibility of 1H and 31P MRS was investigated. The transverse relaxation times for phosphoethanolamine (PE) and phosphocholine (PC) were fitted bi‐exponentially, with an added short T 2 component of 20 ms for adenosine monophosphate, resulting in values of 199 ± 8 and 239 ± 14 ms, respectively. The transverse relaxation time for glycerophosphocholine (GPC) was also fitted bi‐exponentially, with an added short T 2 component of 20 ms for glycerophosphatidylethanolamine, which resonates at a similar frequency, resulting in a value of 177 ± 6 ms. Transverse relaxation times for inorganic phosphate, γ‐ATP and glycerophosphatidylcholine mobile phospholipid were fitted mono‐exponentially, resulting in values of 180 ± 4, 19 ± 3 and 20 ± 4 ms, respectively. Coefficients of variation for the duplicate determinations of 1H total choline (tChol) and the 31P metabolites were calculated for the group of volunteers. The reproducibility of inorganic phosphate, the sum of phosphomonoesters and the sum of phosphodiesters with 31P MRS imaging was superior to the reproducibility of 1H MRS for tChol. 1H and 31P data were combined to calculate estimates of the absolute concentrations of PC, GPC and PE in healthy fibroglandular tissue, resulting in upper limits of 0.1, 0.1 and 0.2 mmol/kg of tissue, respectively.

Skeletal muscle alkaline Pi pool is decreased in overweight-to-obese sedentary subjects and relates to mitochondrial capacity and phosphodiester content

Defects in skeletal muscle energy metabolism are indicative of systemic disorders such as obesity or type 2 diabetes. Phosphorus magnetic resonance spectroscopy (31P-MRS), in particularly dynamic 31P-MRS, provides a powerful tool for the non-invasive investigation of muscular oxidative metabolism. The increase in spectral and temporal resolution of 31P-MRS at ultra high fields (i.e., 7T) uncovers new potential for previously implemented techniques, e.g., saturation transfer (ST) or highly resolved static spectra. In this study, we aimed to investigate the differences in muscle metabolism between overweight-to-obese sedentary (Ob/Sed) and lean active (L/Ac) individuals through dynamic, static, and ST 31P-MRS at 7T. In addition, as the dynamic 31P-MRS requires a complex setup and patient exercise, our aim was to identify an alternative technique that might provide a biomarker of oxidative metabolism. The Ob/Sed group exhibited lower mitochondrial capacity, and, in addition, static 31P-MRS also revealed differences in the Pi-to-ATP exchange flux, the alkaline Pi-pool, and glycero-phosphocholine concentrations between the groups. In addition to these differences, we have identified correlations between dynamically measured oxidative flux and static concentrations of the alkaline Pi-pool and glycero-phosphocholine, suggesting the possibility of using high spectral resolution 31P-MRS data, acquired at rest, as a marker of oxidative metabolism.

Perinatal choline deficiency delays brain development and alters metabolite concentrations in the young pig

Objectives: Adequate choline supply during the perinatal period is critical for proper brain formation, when robust neurogenesis and neuronal maturation occur. Therefore, the objective of this study was to examine the impact of perinatal choline status on neurodevelopment.Methods: Sows were fed a choline-deficient (CD) or choline-sufficient (CS) diet during the last half of the gestational period. At 2 days of age, piglets from sows within each prenatal treatment group were further stratified into postnatal treatment groups and provided either a CD or CS milk replacer, resulting in four treatment groups. At 30 days of age, piglets underwent magnetic resonance imaging (MRI) procedures to analyze structural and metabolite differences.Results: Single-voxel spectroscopy (SVS) analysis revealed postnatally CS piglets had higher (P < 0.001) concentrations of glycerophosphocholine–phosphocholine than postnatally CD piglets. Volumetric analysis indicated smaller (P < 0.006) total brain volumes in prenatally CD piglets compared with prenatally CS piglets. Differences (P < 0.05) in the corpus callosum, pons, midbrain, thalamus, and right hippocampus, were observed as larger region-specific volumes proportional to total brain size in prenatally CD piglets compared with CS piglets. Diffusion tensor imaging (DTI) suggested interactions (P < 0.05) between prenatal and postnatal choline status in fractional anisotropy values of the thalamus and right hippocampus. Prenatally CS piglets had lower cerebellar radial diffusivity (P = 0.045) compared with prenatally CD piglets.Discussion: This study demonstrates that prenatal choline deficiency has profound effects by delaying neurodevelopment as evidenced by structural and metabolic MRI assessments.

Metabolic abnormalities of gastrointestinal mucosa in celiac disease: An in vitro proton nuclear magnetic resonance spectroscopy study.

Celiac disease (CeD) is a common autoimmune disorder in which ingestion of gluten and related proteins leads to inflammation in the small intestine. Although the histological findings in CeD are characteristic, they are not specific. In this study, proton nuclear magnetic resonance (NMR) spectroscopy was used to investigate the differences in metabolic profile of duodenal mucosal biopsies of patients with CeD and controls to find out the biomarker/s of villous atrophy. Duodenal mucosal biopsies were collected from 29 CeD patients (mean age 26.2 ± 10.8 years) and 17 controls (mean age 34.1 ± 11.1 years) and were subjected to proton NMR spectroscopy following perchloric acid extraction. Assignment of metabolite resonances was carried out and their concentrations were determined. For comparison between the groups unpaired t-test/Wilcoxon rank sum test was used. Partial least squares-discriminant analysis was performed to study the clustering behavior of the samples from CeD patients and controls using the Unscrambler 10.2 software. Partial least squares-discriminant analysis clearly differentiated CeD patients from controls. Significantly higher concentrations of isoleucine, leucine, aspartate, succinate, and pyruvate, and lower concentration of glycerophosphocholine, were observed in the duodenal mucosa of CeD patients compared with controls. The results suggest abnormalities in glycolysis, Krebs cycle (energy deficiency), and amino acid metabolism, which may affect the biosynthetic pathways and consequently contribute to villous atrophy. NMR spectroscopy with multivariate analysis of duodenal mucosal biopsies revealed a characteristic metabolic profile in CeD patients. The work provided an insight in determining biomarker/s for villous atrophy and diagnosis of CeD patients.

Altered phospholipid metabolism in schizophrenia: A phosphorus 31 nuclear magnetic resonance spectroscopy study

Phospholipid (PL) metabolism is investigated by in vivo 31P magnetic resonance spectroscopy (MRS). Inconsistent alterations of phosphocholine (PC), phosphoethanolamine (PE), glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE) have been described in schizophrenia, which might be overcome by specific editing techniques. The selective refocused insensitive nuclei-enhanced polarization transfer (RINEPT) technique was applied in a cross-sectional study involving 11 schizophrenia spectrum disorder patients (SZP) on stable antipsychotic monotherapy and 15 matched control subjects. Metabolite signals were found to be modulated by cerebrospinal fluid (CSF) content and gray matter/brain matter ratio. Corrected metabolite concentrations of PC, GPC and PE differed between patients and controls in both subcortical and cortical regions, whereas antipsychotic medication exerted only small effects. Significant correlations were found between the severity of clinical symptoms and the assessed signals. In particular, psychotic symptoms correlated with PC levels in the cerebral cortex, depression with PC levels in the cerebellum and executive functioning with GPC in the insular and temporal cortices. In conclusion, after controlling for age and tissue composition, this investigation revealed alterations of metabolite levels in SZP and correlations with clinical properties. RINEPT 31P MRS should also be applied to at-risk-mental-state patients as well as drug-naïve and chronically treated schizophrenic patients in order to enhance the understanding of longitudinal alterations of PL metabolism in schizophrenia.

Relations of Human Breastmilk Choline Content with Maternal Hormonal Status

An adequate supply of choline is important during the perinatal period for normal growth and neuronal development. Breastmilk is the main nutritional source of choline for breastfed infants. Concentrations of free choline and choline compounds in human breastmilk are influenced by maternal circulatory choline status, maternal diet, and genotype. In the present study, we assessed the relationship between milk content of choline compounds and maternal serum hormonal status. In total, 185 breastfeeding women participated in the study. They were enrolled in the first 3 days after giving birth (expressing colostrum, n = 67), Days 6–14 (expressing transitional milk, n = 47), or Days 28–178 (expressing mature milk, n = 71) postpartum. Written informed consents were obtained from all subjects before participating in the study. The study protocol was approved by the Ethical Committee of Uludag University Medical School (Bursa, Turkey). Foremilk samples were collected by hand expression 3 hours after breakfast and prior to the midday feeding, right after collection of blood samples. Choline compounds were extracted from whole milk and assayed as described previously. Serum hormones were measured by using either the Architect 8200 (Abbott, North Chicago, IL) or the Immulite 2000 (Diagnostic Products Corp., a division of Siemens Healthcare, Los Angeles, CA) autoanalyzer. Commercially available assay kits were used to measure serum leptin, resistin (DSL, Webster, TX), and adiponectin (Linco Research, St. Charles, MO). Relationships between concentrations of choline compounds in milk and hormones in serum were determined by Pearson’s correlation analysis. Values of p< 0.05 were considered significant. Concentrations and content of choline compounds in breastmilk varied in a wide range at all lactation stages and among breastfeeding women (Fig. 1). Colostrum free choline concentrations correlated positively with serum prolactin (r=0.688) and adiponectin (r=0.297) and negatively with serum progesterone (r= -0.336), cortisol (r= -0.390), and leptin (r= -0.353). Phosphocholine concentrations in colostrum correlated positively with serum prolactin (r=0.636), adiponectin (r= 0.333), and resistin (r= 0.248) and negatively with serum cortisol (r= -0.467) and leptin (r= -0.264). Colostrum glycerophosphocholine concentrations correlated positively with serum prolactin (r=0.843) and adiponectin (r=0.264) and negatively with serum progesterone (r= -0.339), cortisol (r= -0.459), and leptin (r= -0.279). Phospholipid-bound choline concentration in colostrum showed weak positive association with serum estradiol (r=0.240) and cortisol (r=0.240) and negative association with adiponectin (r= -0.257). Total choline content correlated positively with serum prolactin (r=0.845) and adiponectin (r=0.278) and negatively with serum progesterone (r= -0.299), cortisol (r= -0.462), and leptin (r= -0.337). Transitional milk free choline concentrations correlated positively with serum prolactin (r= 0.474) and negatively with serum cortisol (r= 0.653) concentrations. Phosphocholine concentrations in transitional milk correlated negatively with serum prolactin (r= 0.334), progesterone (r= 0.311), and cortisol (r= 0.653) concentrations. Transitional milk glycerophosphocholine concentrations correlated negatively with serum prolactin (r= 0.369), progesterone (r= 0.336), and cortisol (r= 0.546) concentrations. Total choline contents in transitional milk correlated negatively with serum prolactin (r= 0.315), progesterone (r= 0.350), and cortisol (r= 0.672) concentrations. No significant correlation was found between concentrations of choline compounds in transitional milk and serum concentrations of estradiol, insulin, thyroid hormones, leptin, adiponectin, resistin, or ghrelin. Mature milk free choline concentrations correlated positively with serum prolactin (r= 0.643) and negatively with serum estradiol (r= 0.254) and cortisol (r= 0.565) concentrations. Phosphocholine concentrations in mature milk correlated negatively with maternal serum estradiol (r= 0.443), progesterone (r= 0.269), cortisol (r= 0332), and insulin (r= 0.406) concentrations. Mature milk glycerophosphocholine concentrations correlated negatively with maternal serum estradiol (r= 0.465), progesterone (r= 0.255), cortisol (r= 0.531), and insulin (r= 0.505) concentrations. Mature milk phospholipid-bound choline concentrations showed negative correlation with serum insulin (r = 0.434) and triiodothyronine (r= 0.310) concentrations. Mature milk total choline concentrations correlated negatively with maternal serum estradiol (r= 0.477), progesterone (r= 0.332), cortisol (r = 0.571), and insulin (r= 0.462) concentrations. No significant correlation was found between concentrations of choline compounds in mature milk and maternal serum concentrations of leptin, adiponectin, resistin, or ghrelin. These data confirm and extend the findings in our previous studies that breastmilk contents of important bioactive

Efficacy of Withania somnifera on seminal plasma metabolites of infertile males: A proton NMR study at 800 MHz

Ethnopharmacological relevanceTraditional Indian systems of medicine use roots of Withania somnifera for impotence, infertility treatment, stress, and the aging process. Although Withania somnifera improves semen quality by regulating reproductive hormone levels and oxidative stress, the molecular mechanism is not clear. Aim of the studyOur study uses high-resolution Nuclear Magnetic Resonance (NMR) spectroscopy to explore the scientific basis to reveal the pre- and post-treatment efficacy of Withania somnifera on seminal plasma of infertile men—which remains unexplored to date. Materials and methodsA total of 180 infertile male patients were administered Withania somnifera root powder at the rate of 5g/d for a 3-month period. The study included age-matched, healthy men as a control (n=50) group. Proton NMR spectroscopy was used to measure lactate, alanine, glutamate, glutamine, citrate, lysine, choline, glycerophosphocholine (GPC), glycine, tyrosine, histidine, phenylalanine, and uridine in all seminal plasma samples. To appraise infertility levels, we also measured sperm concentration, motility, lipid peroxide, and hormonal perturbation. ResultsWithania somnifera therapy repairs the disturbed concentrations of lactate, alanine, citrate, GPC, histidine, and phenylalanine in seminal plasma and recovers the quality of semen of post-treated compared to pre-treated infertile men. Serum biochemistry was also improved over post-therapy in infertile men. Our findings reveal that Withania somnifera not only reboots enzymatic activity of metabolic pathways and energy metabolism but also invigorates the harmonic balance of seminal plasma metabolites and reproductive hormones in infertile men. ConclusionThe results suggest that Withania somnifera may be used as an empirical therapy for clinical management and treatment of infertility.

HDAC Inhibition Induces Increased Choline Uptake and Elevated Phosphocholine Levels in MCF7 Breast Cancer Cells

Histone deacetylase (HDAC) inhibitors have emerged as effective antineoplastic agents in the clinic. Studies from our lab and others have reported that magnetic resonance spectroscopy (MRS)-detectable phosphocholine (PC) is elevated following SAHA treatment, providing a potential noninvasive biomarker of response. Typically, elevated PC is associated with cancer while a decrease in PC accompanies response to antineoplastic treatment. The goal of this study was therefore to elucidate the underlying biochemical mechanism by which HDAC inhibition leads to elevated PC. We investigated the effect of SAHA on MCF-7 breast cancer cells using 13C MRS to monitor [1,2-13C] choline uptake and phosphorylation to PC. We found that PC synthesis was significantly higher in treated cells, representing 154±19% of control. This was within standard deviation of the increase in total PC levels detected by 31P MRS (129±7% of control). Furthermore, cellular choline kinase activity was elevated (177±31%), while cytidylyltransferase activity was unchanged. Expression of the intermediate-affinity choline transporter SLC44A1 and choline kinase α increased (144% and 161%, respectively) relative to control, as determined by mRNA microarray analysis with protein-level confirmation by Western blotting. Taken together, our findings indicate that the increase in PC levels following SAHA treatment results from its elevated synthesis. Additionally, the concentration of glycerophosphocholine (GPC) increased significantly with treatment to 210±45%. This is likely due to the upregulated expression of several phospholipase A2 (PLA2) isoforms, resulting in increased PLA2 activity (162±18%) in SAHA-treated cells. Importantly, the levels of total choline (tCho)-containing metabolites, comprised of choline, PC and GPC, are readily detectable clinically using 1H MRS. Our findings thus provide an important step in validating clinically translatable non-invasive imaging methods for follow-up diagnostics of HDAC inhibitor treatment.