Low Choline Research Articles

Distribution of functional 1‐carbon metabolism single nucleotide polymorphisms in Gambian versus European populations (827.4)

Choline is an essential dietary nutrient and, along with its metabolites, has profound implications in cell signaling, methyl group donation, neurotransmitter synthesis, structural integrity of cell membranes, and fetal neurogenesis. Several important single nucleotide polymorphisms (SNPs) in the choline and folate metabolism pathways have been associated with organ dysfunction, either liver or muscle damage, in response to a low choline diet. A study was conducted to compare the distribution of these known functional SNPs in pregnant women in rural Gambia (N=248), a population with a low choline diet, versus a North Carolina population of European Descent (N=230), a population with a relatively high choline diet. The effect alleles of these SNPs increase dietary choline requirements and contribute to organ dysfunction. The prevalence of the effect allele of functional SNPs in 1‐carbon metabolism was significantly different in Gambian versus European populations: PEMT rs12325817 2.4% and 17.5%, MTHFD1 rs2236225 2.4% and 15.0 %; CHDH rs12676 0.0% and 9.4 % (P<0.0001). Interestingly, in the Gambian population, the effect alleles are reduced or absent as compared to the North Carolina population. This supports our current hypothesis that dietary choline intake is a novel selective pressure that could lead to the differences in SNP distribution and associated risk for organ dysfunction in these populations.Grant Funding Source: Supported by The Gates Foundation

Correlation between dietary intake and plasma levels of choline and betaine in children with autism (827.1)

Alterations in nutrient metabolism may exist in children with autism spectrum disorders (ASD’s), especially the pathways involving folate‐dependent one‐carbon metabolism. Few studies have investigated intake and plasma levels of choline and betaine in children with ASD, although they play a role in these metabolic pathways. This study examined 1) differences in plasma levels of choline and betaine in children with and without ASD and 2) relationships between dietary intake and plasma levels of choline and betaine in children with ASD. Secondary data analyses on 63 children were conducted. Significantly lower choline levels were found in children with ASD (M=8.00, SD=2.42) compared to children without ASD, M=10.03, SD=2.84; t (61) =‐3.04, p= 0.004, (two‐tailed). Similarly, betaine levels were significantly lower in children with ASD (M=24.1, SD=6.49) compared to children without ASD, M=36.70, SD= 6.73; t (60)= ‐7.5, p < 0.001, (two‐tailed). Strong, positive correlations were found between dietary intake and plasma levels of choline (r= 0.657, n=33, p<0.0005) and betaine (r=0.688, n=33, p<0.0005) in children with ASD. These results suggest that children with ASD may have diets inadequate in choline.Grant Funding Source: HRSA; NICHD and Rochester University

A Study of Prognostic Predictors in Organophosphate Poisoning in a Critical Care Unit of a Medical College of Andhra Pradesh

Background : Diagnosis of organophosphate poisoning is purely clinical, through some biochemical test will help in diagnosis of organophosphate poisoning. There is no specific prognostic predictor in terms of duration of hospital stay and course of recovery. Good prognostic predictor can guide the clinician for further treatment. Objectives : 1. To study the prognostic predictor with special reference to duration of the hospital stay. 2. To predict duration of hospital stay from initial clinical sign and biochemical parameters. Materials and method : Around 240 patients of organophosphate poisoning who were admitted in the critical care unit in the hospital from January 2007 to January 2010 were examined, investigated and interview with help of semi structured proforma. The initial clinical presentation and biochemical parameter were recorded for the study. Result: Patients who presented clinically with severe symptoms at the time of admission, increased leucocytes counts, electrolytes and low serum acetyl choline esterase level, those patients stayed more days in ICU and most of the patients required ventilator supports. Conclusio n: It is very important to predict and select the case by detailed clinical history and biochemical examination, so that better treatment and reduction of the mortality can be achieved.

Mo1002 In Non-Alcoholic Fatty Liver Disease, Differential Gene Expression for Phospholipid Metabolism Is Associated With Lower Hepatic Phosphatidylcholine to Phosphatidylethanolamine Ratio, but Not With Lower Choline Intake

Mo1002 In Non-Alcoholic Fatty Liver Disease, Differential Gene Expression for Phospholipid Metabolism Is Associated With Lower Hepatic Phosphatidylcholine to Phosphatidylethanolamine Ratio, but Not With Lower Choline Intake

Choline Modulates Soluble‐Endoglin (sEng) and Vascular Endothelial Growth Factor (VEGF) in Cultured Human Placental Trophoblasts

Altering the balance between the anti‐angiogenic factors, soluble fms‐related tyrosine kinase‐1 (sFLT1) and sEng, and the angiogenic factors, VEGF and transforming growth factor beta (TGF‐β), contributes to preeclampsia (PE) and persistence of hypoxia (Hx). Increasing sFLT1 and sEng may sequester VEGF and TGF‐β. Recently we showed that supplemental choline reduced placental sFLT1 mRNA and maternal plasma sFLT1 in pregnant women and cultured trophoblasts (HTR‐8/SVneo). To determine if choline regulates sEng production and VEGF gene expression independently or interactively with Hx, HTR‐8/SVneo cells were cultured in a 2×4 factorial design in either normoxia (20% O2, Nx) or Hx (1% O2) and 13 μM, 18 μM, 28 μM (control) or 48 μM for 96 hours. Results were analyzed by 2‐way ANOVA (SigmaPlot). Low choline (13 μM) under Nx, but not Hx, increased media sEng (14%, p=0.036) and mRNA levels of VEGF (160.9%, p<0.001), HIF‐1α (36.2%, p<0.001) and HIF‐2α (63.3%, p<0.001) compared to control. Hx also increased media sEng (9.25%, p<0.001) and VEGF mRNA levels (114.18%, p<0.001) across all choline levels. Thus, low choline modulates, under Nx but not Hx, the anti‐angiogenic sEng as well as the angiogenic VEGF, thereby mimicking the effects of Hx. Low choline under Nx, therefore, may alter the resulting balance of antiangiogenic and angiogenic factors contributing to PE. Funded by USDA NIFA AFRI003397

Choline affects placental vascular function and development in a dose response manner

This study explored the role of choline in placental vascular function. In a 12‐week controlled feeding study among third trimester pregnant women, the higher choline intake group (930 vs. 480 mg/d) had lower (P=0.05) placental and circulating levels of soluble fms‐like tyrosine kinase‐1 (sFLT1), an anti‐angiogenic factor that predicts preeclampsia risk. The effect of choline on placental cell function was further investigated using immortalized HTR‐8/SVneo trophoblasts cultured in different choline concentrations (28, 13 and 8 μM) for 96 hours. The decrease in choline concentrations led to a graded increase (P<0.01) in the gene expression and secretion of sFLT1, expression of the proinflammatory markers [e.g. interleukin 1 beta (IL1B)], apoptosis and oxidative stress. Cellular phosphatidylcholine dramatically decreased (50%, P<0.01) in the lower choline group, whereas the protein kinase C (PKC) isoforms δ and ε, activated by breakdown products of phosphatidylcholine, increased (P<0.01). The addition of a PKC inhibitor partially rescued the increased expression of sFLT1 and apoptosis. In conclusion, choline influences placental vascular function partially via PKC.Grant Funding Source: USDA NIFA AFRI003397

Brain Magnetic Resonance Spectroscopy in Episodic Hepatic Encephalopathy

Brain magnetic resonance (MR) study has shown metabolic abnormalities and changes in water distribution of the brain tissue that may relate to the pathogenesis of hepatic encephalopathy (HE). We designed a study to investigate the disturbances in brain water and metabolites during episodic HE using a 3-T MR scanner. Cirrhotic patients with different grades of HE underwent MR during hospitalization (n=18). The MR was repeated at 6 weeks' follow-up (n=14). The results were compared with those of a group of healthy volunteers (n=8). During episodic HE, brain diffusion-weighted imaging showed a high apparent diffusion coefficient (ADC) (12% to 14%) that decreased during follow-up (-1% to -4%). These disturbances were accompanied by high glutamine (581%), low choline (-31%), and low myo-inositol (-86%) peaks on MR spectroscopy. In overt HE, patients showed high glutamine that decreased during follow-up (-22%). In addition, these patients exhibited a rise in plasma S100 beta and enlargement of brain white-matter lesions. In conclusion, several disturbances detected by MR support the presence of impaired brain water homeostasis during episodic HE. Although astrocytes have a major role in this condition, brain edema during episodic HE may be extracellular and does not appear to be directly responsible for the development of neurologic manifestations.

Identification and Characterization of a High-Affinity Choline Uptake System of Brucella abortus

Phosphatidylcholine (PC), a common phospholipid of the eukaryotic cell membrane, is present in the cell envelope of the intracellular pathogen Brucella abortus, the etiological agent of bovine brucellosis. In this pathogen, the biosynthesis of PC proceeds mainly through the phosphatidylcholine synthase pathway; hence, it relies on the presence of choline in the milieu. These observations imply that B. abortus encodes an as-yet-unknown choline uptake system. Taking advantage of the requirement of choline uptake for PC synthesis, we devised a method that allowed us to identify a homologue of ChoX, the high-affinity periplasmic binding protein of the ABC transporter ChoXWV. Disruption of the choX gene completely abrogated PC synthesis at low choline concentrations in the medium, thus indicating that it is a high-affinity transporter needed for PC synthesis via the PC synthase (PCS) pathway. However, the synthesis of PC was restored when the mutant was incubated in media with higher choline concentrations, suggesting the presence of an alternative low-affinity choline uptake activity. By means of a fluorescence-based equilibrium-binding assay and using the kinetics of radiolabeled choline uptake, we show that ChoX binds choline with an extremely high affinity, and we also demonstrate that its activity is inhibited by increasing choline concentrations. Cell infection assays indicate that ChoX activity is required during the first phase of B. abortus intracellular traffic, suggesting that choline concentrations in the early and intermediate Brucella-containing vacuoles are limited. Altogether, these results suggest that choline transport and PC synthesis are strictly regulated in B. abortus.

Effects of choline on sodium arsenite-induced neural tube defects in chick embryos

Arsenic passes through the placenta and accumulates in the neuroepithelium of embryo, whereby inducing congenital malformations such as neural tube defects (NTDs) in animals. Choline (CHO), a methyl-rich nutrient, functions as a methyl donor to participate in methyl group metabolism. Arsenic methylation has been regarded as a detoxification process and choline (CHO) is the major source of methyl-groups. However, whether CHO intake reverses the abnormal embryo development induced by sodium arsenite (SA) and the relationship between CHO intake and arsenite-induced NTDs are still unclear. In this study, we used chick embryos as animal model to investigate the effects of SA and CHO supplementation on the early development of nervous system. Our results showed that the administration of SA led to reduction in embryo viability, embryo body weight and extraembryonic vascular area, accompanied by a significantly increased incidence of the failed closure of the caudal end of the neural tube. CHO, at low dose (25μg/μL), reversed the decrease in embryo viability and the increase in the failed closure of the caudal end of the neural tube, which were induced by SA. In addition, CHO (25μg/μL) inhibited not only the SA-induced cell apoptosis by up-regulating Bcl-2 level, but also the global DNA methylation by increasing the expressions of DNMT1 and DNMT3a. However, less significant difference was found between the embryos co-treated with SA and CHO (50μg/μL) and the ones treated with SA alone. Taken together, these findings suggest that low dose CHO could protect chick embryos from arsenite-induced NTDs by a possible mechanism related to the methyl metabolism.

Plasma free choline, betaine and cognitive performance: the Hordaland Health Study

Choline and betaine are nutrients involved in one-carbon metabolism. Choline is essential for neurodevelopment and brain function. We studied the associations between cognitive function and plasma concentrations of free choline and betaine. In a cross-sectional study, 2195 subjects (55 % women), aged 70-74 years, underwent extensive cognitive testing including the Kendrick Object Learning Test (KOLT), Trail Making Test (part A, TMT-A), modified versions of the Digit Symbol Test (m-DST), Block Design (m-BD), Mini-Mental State Examination (m-MMSE) and Controlled Oral Word Association Test (COWAT). Compared with low concentrations, high choline (>8·4 μmol/l) was associated with better test scores in the TMT-A (56·0 v. 61·5, P=0·004), m-DST (10·5 v. 9·8, P=0·005) and m-MMSE (11·5 v. 11·4, P=0·01). A generalised additive regression model showed a positive dose-response relationship between the m-MMSE and choline (P=0·012 from a corresponding linear regression model). Betaine was associated with the KOLT, TMT-A and COWAT, but after adjustments for potential confounders, the associations lost significance. Risk ratios (RR) for poor test performance roughly tripled when low choline was combined with either low plasma vitamin B₁₂ (≤257 pmol/l) concentrations (RR(KOLT)=2·6, 95 % CI 1·1, 6·1; RR(m-MMSE)=2·7, 95 % CI 1·1, 6·6; RR(COWAT)=3·1, 95 % CI 1·4, 7·2) or high methylmalonic acid (MMA) (≥3·95 μmol/l) concentrations (RR(m-BD)=2·8, 95 % CI 1·3, 6·1). Low betaine (≤31·1 μmol/l) combined with high MMA was associated with elevated RR on KOLT (RR(KOLT)=2·5, 95 % CI 1·0, 6·2). Low plasma free choline concentrations are associated with poor cognitive performance. There were significant interactions between low choline or betaine and low vitamin B₁₂ or high MMA on cognitive performance.

Altered Expression of Alzheimer's Disease-Related Proteins in Male Hypogonadal Mice

Age-related depletion of estrogens and androgens is associated with an increase in Alzheimer's disease (AD) brain pathology and diminished cognitive function. Here we investigated AD-associated molecular and cellular changes in brains of aged hypogonadal (hpg) male and female mice. hpg Mice have a spontaneous, inactivating genetic mutation in the GnRH gene resulting in life-long deficiency of gonadotropins and gonadal sex hormones. Western blot analysis revealed low levels of amyloid precursor protein and high levels of presenilin 1, amyloid precursor protein C-terminal fragment, and β-amyloid 42 in brains of aged male, but not female, hpg mice. Changes were confined to the hippocampus and were not evident in the cerebellum or other brain tissues. Male hpg mice tended to have lower levels of IL-1β protein than male littermate controls. Immunohistochemical staining of the basal forebrain revealed that male hpg mice had lower choline acetyltransferase levels per neuron compared with controls. These AD-like changes specific to male hpg mice supports a link between androgen depletion and the development of AD pathology.

Reply to the letter “Choline PET/CT compared with bone scintigraphy in the detection of bone metastases in prostate cancer patients”

Dear Sir, We thank Beheshti and Langsteger [1] for their interest in our paper “[C]Choline PET/CT detection of bone metastases in patients with PSA progression after primary treatment for prostate cancer: comparison with bone scintigraphy” [2] and for raising important issues; we would like to reply to the comments presented. We agree that the comparison between tomographic and planar modalities has an intrinsic bias due to the different potentials of the two techniques. Knowing that it has already been demonstrated that single photon emission computed tomography (SPECT) and/or SPECT/CT significantly improve the specificity of bone scintigraphy (BS) [3, 4], in the present paper the clinical advantages and limitations of one technique over the other have been described exclusively based on a clinical point of view, and the comparison between the two modalities to evaluate the potential benefits that PET/CT could offer over BS have been performed. We are aware that the comparison of bone metastases may be of limited value because [C]choline PET/CTstudies were performed from the base of the skull. This limitation is due to the retrospective nature of the study; thus, [C]choline PET/ CT scans were acquired according to a standard protocol. In particular, only two lesions, one at the skull, and categorized at BS as malignant, and one at an arm, and categorized at BS as equivocal, could not be validated because the anatomical localizations were outside the field of view of PET/CT and thus excluded from lesion-based analysis. Regarding the low choline uptake in sclerotic bone lesions, our opinion is that further studies are required to clarify the meaning of this phenomenon. At present we cannot distinguish between the low uptake due to a regression of bone disease in response to therapy or due to a false-negative [C] choline PET/CT finding. Until this crucial concept has been clarified, the monitoring of bone metastatic disease should be based on a conventional bone scan. At present the low choline uptake in sclerotic bone lesions can be considered a limitation of [C]choline PET/CT but, once the metabolic meaning of this finding is clarified, it could potentially become a prognostic tool with significant clinical relevance. In our series, there were 24 of 78 patients with bone metastases on [C]choline PET/CT and 19 of 78 patients on BS. In our opinion the relevance of these results is the following: if a patient undergoes [C]choline PET/CT for biochemical recurrence for restaging his disease and a pathological bone uptake occurs, due to the high positive predictive value of [C]choline PET/CT, the clinician may be confident in considering that pathological uptake as a bone metastatic lesion. Moreover, the absence of [C]choline uptake in degenerative changes of the skeleton is another issue that emphasizes the potential superiority of this technique over BS. In response to the comment of Beheshti and Langsteger [1], we would also like to remark that [C]choline PET/CT, being a whole-body modality, presents the great advantage M. Picchio (*) : F. Fallanca Nuclear Medicine Department, Scientific Institute San Raffaele, Milan, Italy e-mail: picchio.maria@hsr.it

Choline supplementation and measures of choline and betaine status: a randomised, controlled trial in postmenopausal women

Choline is an essential nutrient and can also be obtained by de novo synthesis via an oestrogen responsive pathway. Choline can be oxidised to the methyl donor betaine, with short-term supplementation reported to lower plasma total homocysteine (tHcy); however, the effects of longer-term choline supplementation are less clear. We investigated the effect of choline supplementation on plasma concentrations of free choline, betaine and tHcy and B-vitamin status in postmenopausal women, a group more susceptible to low choline status. We also assessed whether supplementation altered plasma lipid profiles. In this randomised, double-blinded, placebo-controlled study, forty-two healthy postmenopausal women received 1 g choline per d (as choline bitartrate), or an identical placebo supplement with their habitual diet. Fasting blood samples were collected at baseline, week 6 and week 12. Administration of choline increased median choline and betaine concentrations in plasma, with significant effects evident after 6 weeks of supplementation (P<0·001) and remaining significant at 12 weeks (P<0·001); no effect was observed on folate status or on plasma lipids. Choline supplementation induced a median (25th, 75th percentile) change in plasma tHcy concentration at week 6 of -0·9 (-1·6, 0·2) μmol, a change which, when compared to that observed in the placebo group 0·6 (-0·4, 1·9) μmol, approached statistical significance (P=0·058). Choline supplementation at a dose of 1 g/d significantly increases the circulating concentration of free choline, and can also significantly increase the concentration of the methyl donor, betaine, thereby potentially enhancing the betaine-homocysteine methyltransferase-mediated remethylation of tHcy.

Prognostic relevance of oncological serum biomarkers in liver cancer patients undergoing transarterial chemoembolization therapy

As transarterial chemoembolization (TACE) therapy is an effective locoregional treatment for patients with advanced liver cancer, prognostic biomarkers are highly needed for pretherapeutic stratification of patients to TACE therapy. Sera of 50 prospectively and consecutively included patients with hepatocellular carcinoma (HCC) undergoing TACE were taken before and 24 h after TACE application. Levels of liver-specific, tumor-related, and cell death biomarkers were analyzed and correlated with overall patient survival. The study was particularly focused on patients treated by TACE with palliative intention (N = 38). Sixteen of 38 patients died within 1 year after TACE, 22 were still alive. In univariate analysis, high levels of cytokeratin 19-fragments (CYFRA 21-1), alpha fetoprotein (AFP), and low choline esterase (CHE) levels measured before and 24 h after TACE were correlated with unfavorable outcome. Further high pretherapeutic lactate dehydrogenase (LDH), aspartate-aminotransferase, and bilirubin levels as well as high 24 h C-reactive protein values were associated with poor survival. In multivariate analysis of clinical and only pretherapeutic biomarkers, AFP, CHE, and LDH showed to be independent prognostic parameters. When additionally 24 h values were included, CHE (24 h) and AFP (24 h) were the strongest independent prognostic biomarkers with a slightly higher prognostic power (Akaike's information criterion 90.3 vs. 92.7). The combination of AFP, CHE, and LDH enables efficient pretherapeutic stratification of HCC patients in advanced tumor stage for TACE therapy.

In vivo MRS of locally advanced breast cancer: characteristics related to negative or positive choline detection and early monitoring of treatment response

To explore factors determining the detection of total choline (tCho) by in vivo MR spectroscopy (MRS) in locally advanced breast cancer and to evaluate the ability of in vivo tCho to predict treatment response after one cycle of neoadjuvant chemotherapy (NAC). Breast cancer patients (N=40) scheduled for NAC were examined with an MR protocol including in vivo single voxel proton MRS, dynamic contrast enhanced MRI and diffusion weighted MRI. tCho was quantified based on the signal-to-noise ratio. The detection was considered positive with tCho signal-to-noise ratio≥2. tCho was detected in 60% of the patients. Excellent reliability in tCho (ICC=0.97, P<0.001) measurements was confirmed. The water/fat-ratio, tumor volume and distribution of Type II voxels (based on contrast uptake curve) were significantly higher in patients with positive choline detection. The probability of detecting tCho was higher in ER-negative patients. A significant decrease in tChoSNR was detected after treatment, but responders could not be distinguished from non-responders. The use of in vivo MRS in breast cancer diagnosis and treatment monitoring should bring supplementary information to the standard MR imaging protocol. With the currently observed low choline detection rate, this is not the case, and technological challenges related to choline detection have to be resolved.

Effects of choline on sodium arsenite-induced neural tube defects in chick embryos

Arsenic passes through the placenta and accumulates in the neuroepithelium of embryo, whereby inducing congenital malformations such as neural tube defects (NTDs) in animals. Choline (CHO), a methyl-rich nutrient, functions as a methyl donor to participate in methyl group metabolism. Arsenic methylation has been regarded as a detoxification process and choline (CHO) is the major source of methyl-groups. However, whether CHO intake reverses the abnormal embryo development induced by sodium arsenite (SA) and the relationship between CHO intake and arsenite-induced NTDs are still unclear. In this study, we used chick embryos as animal model to investigate the effects of SA and CHO supplementation on the early development of nervous system. Our results showed that the administration of SA led to reduction in embryo viability, embryo body weight and extra-embryonic vascular area, accompanied by a significantly increased incidence of the failed closure of the caudal end of the neural tube. CHO, at low dose (25 μg/μL), reversed the decrease in embryo viability and the increase in the failed closure of the caudal end of the neural tube, which were induced by SA. In addition, CHO (25 μg/μL) inhibited not only the SA-induced cell apoptosis by up-regulating Bcl-2 level, but also the global DNA methylation by increasing the expressions of DNMT1 and DNMT3a. However, less significant difference was found between the embryos co-treated with SA and CHO (50 μg/μL) and the ones treated with SA alone. Taken together, these findings suggest that low dose CHO could protect chick embryos from arsenite-induced NTDs by a possible mechanism related to the methyl metabolism.

In Vivo Detection of Choline in Ovarian Tumors Using 3D Magnetic Resonance Spectroscopy

To assess the clinical feasibility of 3-dimensional (3D) proton magnetic resonance spectroscopy (MRS) of ovarian masses at 1.5 T. We prospectively evaluated 16 patients with 23 ovarian masses using contrast-enhanced magnetic resonance imaging and 3D chemical shift imaging MRS (time of reception/time of echo = 700/135 ms, number of excitations = 6, interpolated voxel = 5 × 5 × 5 mm(3), water and fat suppression). Spectral editing consisted of water reference, filtering, zero-filling, Fourier transformation, frequency shift, automatic baseline and phase correction, and curve fitting. The volume of interest was placed to encompass both solid and cystic tumor components as well as apparently healthy pelvic tissues. The presence of a choline peak at 3.14 to 3.34 ppm was considered as a marker of malignancy. All patients underwent surgery and histopathological evaluation. Of 23 masses, 19 were malignant and the remaining 4 benign lesions were a fibrothecoma, an endometriosis, a cyst, and a cystadenofibroma. A choline peak was detected in 17/19 malignant tumors (sensitivity 89%), absent in 2 G1 tumors. It was visible in 16 solid components of 19 malignant tumors (in one of them, a choline peak was detected only in the cystic component, in 6 in both solid and cystic components). The choline peak was absent in 20/21 apparently healthy pelvic tissues, with a very low choline peak being detected in one intraperitoneal fluid collection with malignant cells at cytologic analysis; 3/4 benign tumors showed a choline peak (overall specificity 21/25 = 84%). A significant difference between the mean choline peak integral detected within the solid component and that within the cystic component was observed (P = 0.002). No correlation between the choline peak integral and the tumor size was found (r = 0.120, P = 0.615). 3D MRS of ovarian masses is clinically feasible at 1.5 T. This opens new research strategies for early diagnosis of ovarian cancer.

Maternal dietary choline deficiency alters angiogenesis in fetal mouse hippocampus

We examined whether maternal dietary choline modulates angiogenesis in fetal brain. Pregnant C57BL/6 mice were fed either a choline‐deficient (CD), control (CT), or choline‐supplemented diet (CS) from days 12 to 17 (E12‐17) of pregnancy and then fetal brains were studied. In CD fetal hippocampus, proliferation of endothelial cells (EC) was decreased by 32% (p &lt; 0.01 vs. CT or CS) while differentiated EC clusters (expressing factor VIII related antigen (RA)) increased by 25% (p &lt; 0.01 vs. CT or CS). These changes were associated with &gt; 25% decrease in the number of blood vessels in CD fetal hippocampus (p &lt; 0.01 vs. CT and CS), with no change in total cross‐sectional area of these blood vessels. Expression of genes for the angiogenic signals derived from both endothelial and neuronal progenitor cells (NPC) was increased in CD fetal hippocampus VEGF C (Vegfc), 2.0‐fold, p &lt; 0.01 vs. CT and angiopoietin 2 (Angpt2), 2.1‐fold, (p &lt; 0.01 vs. CT)). Similar increased expression was observed in NPC isolated from E14 fetal mouse brains and exposed to low (5 μM), CT (70 μM), or high choline (280 μM) media for 72 h (low choline caused a 9.7‐fold increase in relative gene expression of Vegfc (p &lt; 0.001 vs. CT and high) and a 3.4‐fold increase in expression of Angpt2, (p &lt; 0.05 vs. CT and high). ANGPT2 protein was increased 42.2% (p &lt; 0.01). Cytosine‐phosphate‐guanine dinucleotide islands in the proximity of the promoter areas of Vegfc and Angpt2 were hypomethylated in low choline NPC compared to CT NPC (p &lt; 0.01). We conclude that maternal dietary choline intake alters angiogenesis in the developing fetal hippocampus. This work was supported by Grants from the National Institutes of Health AG09525, DK55865, and DK56350 (to S.H.Z.) and Pilot Grant ES010126‐08 (to M.G.M.)

P.3.006 Substance use and regional grey matter volume in individuals at high risk of psychosis

Proton magnetic resonance spectroscopy (MRspec), one of the very few techniques for in vivo assessment of neuro-metabolic profiles, is often complicated by lack of standard population norms and paucity of computational tools.7035 scans and clinical information from 4430 pediatric patients were collected from 2008 to 2014. Scans were conducted using a 1.5 T (n = 3664) or 3 T scanner (n = 3371), and with either a long (144 ms, n = 5559) or short echo time (35 ms, n = 1476). 3055 of these scans were localized in the basal ganglia (BG), 1211 in parieto-occipital white matter (WM). 34 metabolites were quantified using LCModel. A web application using MySQL, Python and Flask was developed to facilitate the exploration of the data set.Already piloting the application revealed numerous insights. (1), N-acetylaspartate (NAA) increased throughout all ages. During early infancy, total choline was highly varied and myo-inositol demonstrated a downward trend. (2), Total creatine (tCr) and creatine increased throughout childhood and adolescence, though phosphocreatine (PCr) remained constant beyond 200 days. (3), tCr was higher in BG than WM. (4), No obvious gender-related differences were observed. (5), Field strength affects quantification using LCModel for some metabolites, most prominently for tCr and total NAA. (6), Outlier analysis identified patients treated with vigabatrin through elevated γ-aminobutyrate, and patients with Klippel-Feil syndrome, Leigh disease and L2-hydroxyglutaric aciduria through low choline in BG.We have established the largest MRSpec database and developed a robust and flexible computational tool for facilitating the exploration of vast metabolite datasets that proved its value for discovering neurochemical trends for clinical diagnosis, treatment monitoring, and research. Open access will lead to its widespread use, improving the diagnostic yield and contributing to better understanding of metabolic processes and conditions in the brain.

Use of canonical variate analysis biplot in examination of choline content data of some foods

Adequate intake (AI) of choline as part of the daily diet can help prevent major diseases. Low choline intake is a major risk factor for liver and several neurological disorders. Extreme choline consumption may cause diseases such as hypotension, sweating, diarrhea, and fishy body odor. The AI of choline is 425 mg/day for adult women; higher for pregnant and lactating women. The AI for adult men is 550 mg/day. The total choline content of foods is calculated as the sum of free choline, glycerophosphocholine, phosphocholine, phosphatidylcholine and sphingomyelin. These are called the choline variables. Observed values of choline variables may be different in amounts of nutrients. So different food groups in terms of choline variables are useful to compare. The present paper shows the advantages of using canonical variate analysis biplot to optimally separate groups and explore the differentiality of choline variables amounts in foods.