Data_Sheet_1_Reduction of Glycolysis Intermediate Concentrations in the Cerebrospinal Fluid of Alzheimer’s Disease Patients.docx

The profile of 122 metabolites in the cerebrospinal fluid (CSF) of patients suffering from Alzheimer’s disease (AD) and controls was studied. Among the 122 metabolites analyzed, 61 could be detected. Statistically significant differences between the AD and control group were only detected for metabolites of the glycolysis. Thus, accurate quantification of 11 glycolytic metabolites was done. We detected a significant reduction of five of them, namely phosphoenolpyruvate, 2-phosphoglycerate, 3-phosphoglycerate, pyruvate and dihydroxyacetone phosphate in the AD CSF compared to controls. These results correlate with the known reduction of glucose metabolism in the brain of patients with AD and indicate that metabolic analysis of the central carbon metabolism can be a potential tool in AD diagnostic. Although the Receiver operating characteristic (ROC) analyses of the metabolites do not reach the level of the diagnostic informativity of AD biomarkers, the combination of specific glycolysis metabolites with the established biomarkers may lead to an improvement in sensitivity and specificity.</p

Demographic data and disease characteristics in schizophrenic patients (n = 145) and controls (n = 145); between-group comparisons.

1)<p>dose equivalent to [mg] Chlorpromazine;</p>2)<p>T-test for independent samples (two-sided);</p>3)<p>χ²-Test.</p><p>Significant results are indicated in bold type.</p

Protective effect of creatine in hippocampal cell cultures challenged with oxidative stress.

<p>Hippocampal cells (DIV 15) were incubated with hydrogen peroxide in rising concentrations in absence or in presence of 5 mM creatine. After 24 h the LDH release into the cell culture supernatant was assessed. Total protein of the cell monolayer was used as a reference. Data are expressed as arbitrary units per mg protein +/− standard deviation. Each data point represents the mean of triplicates. Each experiment was independently performed in triplicate. Statistical analysis was performed by unpaired Student's T-test. *denotes statistical significance at a level of p<0.01.</p

Effect of creatine on intracellular ATP/Phosphocreatine content in hippocampal cells exposed to glutamate.

<p>Hippocampal cells (DIV 17) were challenged with glutamate at rising concentrations in absence or presence of 5 mM creatine. After 24 h of incubation the cells were harvested and intracellular ATP/PCr concentration was determined by luciferin/luciferase chemiluminescence. Total protein content of the cell lysate was employed as a reference. Data are expressed as intracellular ATP concentration equivalents corrected for total protein +/− standard deviation. Each data point represents the mean of triplicates. The experiment was independently performed in triplicate. Unpaired Student's T-test was used for statistics. *denotes statistical significance at a level of p<0.01.</p

Impact of creatine on glutamate efflux into the supernatant in hippocampal cell cultures exposed to hydrogen peroxide.

<p>Hippocampal cells (DIV 15) were incubated with rising concentrations of hydrogen peroxide in absence or in presence of 5 mM creatine. After 24 h the glutamate release into the cell culture supernatant was enzymatically determined. Total protein of the lysed cell monolayer was used as a reference. Data are expressed as glutamate concentration per mg protein +/− standard deviation. Each data point represents the mean of triplicates. Each experiment was independently performed in triplicate. Statistical analysis was performed by unpaired Student's T-test. *denotes statistical significance at a level of p<0.01.</p

Effect of creatine on intracellular ATP/Phosphocreatine content in hippocampal cells under oxidative stress.

<p>Hippocampal cells (DIV 15) were challenged with hydrogen peroxide at rising concentrations in absence or presence of 5 mM creatine. After 24 h the cells were harvested for determination of intracellular ATP/PCr concentration, which was determined by luciferin/luciferase chemiluminescence and for measurement of total protein content, which served as a reference. Data are expressed as intracellular ATP concentration equivalents corrected for total protein +/− standard deviation. Each data point represents the mean of triplicates. The experiment was independently performed in triplicate. Unpaired Student's T-test was used for statistics. *denotes statistical significance at a level of p<0.01.</p

Protective effect of creatine in hippocampal cell cultures exposed to glutamate.

<p>Hippocampal cells (DIV 17) were incubated with rising concentrations of glutamate in absence or in presence of 5 mM creatine. After 24 h the LDH release into the cell culture supernatant was determined. Total protein of the lysed cell monolayer was used as a reference. Data are expressed as arbitrary units per mg protein +/− standard deviation. Each data point represents the mean of triplicates. Each experiment was independently performed in triplicate. Statistical analysis was performed by unpaired Student's T-test. *denotes statistical significance at a level of p<0.01.</p

Raw data of IRI scores (mean, SD) by OXTR rs2254298 and rs53576 genotypes in schizophrenia patients (SZ, n = 145) and healthy controls (HC, n = 145).

1)<p>T-test for independent samples: T = −3.493, p<0.001.</p

Impact of creatine pre-incubation on NMDA-triggered intracellular calcium rise in hippocampal cells.

<p>Hippocampal cell cultures (DIV 18) were incubated with 5 mM of creatine for 18 h. Cells were harvested, dissociated and loaded with FURA PE-3/AM. Ca²⁺ ratiometry was performed in 0.5×10⁶ cells/ml at 37°C. After stable baseline ratios were achieved NMDA was added and the response was recorded for 400 seconds. Thapsigargin was added for SERCA inhibition. The tracings are representative for 5 individual experiments by calculating curve means. Data for intracellular Ca²⁺ are expressed in arbitrary units. The second tracing shows responses in creatine-pretreated cells, the first one has been acquired from control cells.</p

OXTR rs2254298 and rs53576 polymorphisms and PANSS positive, negative and general psychopathology scores in schizophrenic patients.

<p>Schizophrenic patients carrying AA- or AG-genotypes of OXTR rs2254298 show significantly higher PANSS general psychopathology scores than GG-carriers (n = 145; mean, SD; t-test for independent samples, *: p<0.05).</p