GABA and Glutamate in Children with Primary Complex Motor Stereotypies: An 1H-MRS Study at 7T.

BACKGROUND AND PURPOSE: Complex motor stereotypies are rhythmic, repetitive, fixed, purposeful but purposeless movements that stop with distraction. They can occur in otherwise normal healthy children (primary stereotypies) as well in those with autism spectrum disorders (secondary stereotypies). The underlying neurobiologic basis for these movements is unknown but is thought to involve cortical-striatal-thalamo-cortical pathways. To further clarify potential neurochemical alterations, gamma-aminobutyric acid (GABA), glutamate, glutamine, N-acetylaspartate, and choline levels were measured in 4 frontostriatal regions by using (1)H MRS at 7T. MATERIALS AND METHODS: A total of 18 children with primary complex motor stereotypies and 24 typically developing controls, ages 5-10 years, completed MR spectroscopy at 7T. Single voxel STEAM acquisitions from the anterior cingulate cortex, premotor cortex, dorsolateral prefrontal cortex, and striatum were obtained, and metabolites were quantified with respect to Cr by using LCModel. RESULTS: The 7T scan was well tolerated by all the participants. Compared with the controls, children with complex motor stereotypies had lower levels of GABA in the anterior cingulate cortex (GABA/Cr, P = .049; GABA/Glu, P = .051) and striatum (GABA/Cr, P = .028; GABA/Glu, P = .0037) but not the dorsolateral prefrontal cortex or the premotor cortex. Glutamate, glutamine, NAA, and Cho levels did not differ between groups in any of the aforementioned regions. Within the complex motor stereotypies group, reduced GABA to Cr in the anterior cingulate cortex was significantly associated with greater severity of motor stereotypies (r = -0.59, P = .021). CONCLUSIONS: These results indicate possible GABAergic dysfunction within corticostriatal pathways in children with primary complex motor stereotypies.

Low cerebellar vermis volumes and impaired neuropsychologic performance in children treated for brain tumors and leukemia.

BACKGROUND AND PURPOSE: Injury of the cerebellar vermis may occur in children with brain malignancies. Because the vermis is involved in motor and cognitive functioning, the goal of this prospective longitudinal study was to evaluate treatment-related changes in vermal volumes and neuropsychologic performance in children receiving brain radiation of the cerebellum. MATERIALS AND METHODS: Ten patients (mean age, 11.6 years) and 10 healthy children (mean age, 12.1 years) were examined. Lobar vermal volumes and performance on neuropsychologic tests evaluating motor, visual, verbal, attention, memory, and executive functions were assessed at baseline and at 6-month follow-up visits. RESULTS: At baseline, lower mean vermal volumes and impaired performance on visual-spatial and fine-motor tasks were detected in patients. At 6-month follow-up, further decrease in vermal volumes was detected only in patients with medulloblastoma, who received the largest radiation doses to the entire vermis. The volume decrease was not associated with reduction in neuropsychologic performance compared with baseline. At 6-month follow-up, data from all subjects revealed an association between smaller vermal volumes and slower fine-motor speed and lower visual-spatial skills. CONCLUSIONS: Reduced brain-tissue volumes following radiation have been reported previously in pediatric patients. In this study, lower vermal volumes were detected even earlier, before radiation treatment was initiated or completed. Six months postradiation, vermal volume decreases detected in patients with medulloblastoma were not accompanied by declines in already poor neuropsychologic performance. In addition to radiation, the presence of brain malignancies and preradiation treatment may be important factors affecting cerebellar vermis tissue.

White matter impairment in Rett syndrome: diffusion tensor imaging study with clinical correlations.

BACKGROUND AND PURPOSE: RTT, caused by mutations in the methyl CPG binding protein 2 (MeCP2) gene, is a disorder of neuronal maturation and connections. Our aim was to prospectively examine FA by DTI and correlate this with certain clinical features in patients with RTT. MATERIALS AND METHODS: Thirty-two patients with RTT underwent neurologic assessments and DTI. Thirty-seven age-matched healthy female control subjects were studied for comparison. With use of a 1.5T MR imaging unit, DTI data were acquired, and FA was evaluated to investigate multiple regional tract-specific abnormalities in patients with RTT. RESULTS: In RTT, significant reductions in FA were noted in the genu and splenium of the corpus callosum and external capsule, with regions of significant reductions in the cingulate, internal capsule, posterior thalamic radiation, and frontal white matter. In contrast, FA of visual pathways was similar to control subjects. FA in the superior longitudinal fasciculus, which is associated with speech, was equal to control subjects in patients with preserved speech (phrases and sentences) (P = .542), whereas FA was reduced in those patients who were nonverbal or speaking only single words (P < .001). No correlations between FA values for tracts and clinical features such as seizures, gross or fine motor skills, and head circumference were identified. CONCLUSIONS: DTI, a noninvasive technique to assess white matter tract pathologic features, may add specificity to the assessment of RTT clinical severity that is presently based on the classification of MeCP2 gene mutation and X-inactivation.

Proton MR spectroscopy correlates of frontal lobe function in healthy children.

BACKGROUND AND PURPOSE: Neuroimaging methods have been used to improve our understanding of the topographic organization of the brain. In our study, proton (1)H-MR spectroscopic imaging was used to evaluate frontal lobe function. The goal was to determine the relationship between neuropsychological measures of frontal lobe function and levels of a surrogate neuronal marker, N-acetylaspartate (NAA), in typically developing healthy children and adolescents. MATERIALS AND METHODS: Fifty-one healthy children (25 girls; 6.2-18.3 years of age; mean age, 12.3 +/- 3.6 years) were examined. All children completed a neuropsychological assessment including measures of attention, executive function, memory, language, and visual and motor skills. (1)H-MR spectroscopic imaging was performed by using a multisection spin-echo sequence at 1.5T. General linear model analysis of covariance was used to examine the relationship between the neuropsychological test scores and NAA/creatine (Cr) ratios, controlling for age and sex. RESULTS: A positive relationship between frontal lobe white matter NAA/Cr ratio and performance on 2 neuropsychological tests associated with frontal lobe function was detected. The Purdue Pegboard right-hand scores were higher with increasing NAA/Cr in the left frontal white matter (P = .047), and Stanford-Binet-IV "Bead Memory" scores improved with increasing NAA/Cr ratio in the right frontal white matter (P = .032). CONCLUSIONS: An association between frontal white matter NAA/Cr ratios and 1) measures of manual speed and dexterity, and 2) visual working memory was detected. Our data may provide a quantitative basis for assessment of frontal lobe impairments in disease states.

Quantitative comparison and evaluation of software packages for assessment of abdominal adipose tissue distribution by magnetic resonance imaging.

OBJECTIVE: To examine five available software packages for the assessment of abdominal adipose tissue with magnetic resonance imaging, compare their features and assess the reliability of measurement results. DESIGN: Feature evaluation and test-retest reliability of softwares (NIHImage, SliceOmatic, Analyze, HippoFat and EasyVision) used in manual, semi-automated or automated segmentation of abdominal adipose tissue. SUBJECTS: A random sample of 15 obese adults with type 2 diabetes. MEASUREMENTS: Axial T1-weighted spin echo images centered at vertebral bodies of L2-L3 were acquired at 1.5 T. Five software packages were evaluated (NIHImage, SliceOmatic, Analyze, HippoFat and EasyVision), comparing manual, semi-automated and automated segmentation approaches. Images were segmented into cross-sectional area (CSA), and the areas of visceral (VAT) and subcutaneous adipose tissue (SAT). Ease of learning and use and the design of the graphical user interface (GUI) were rated. Intra-observer accuracy and agreement between the software packages were calculated using intra-class correlation. Intra-class correlation coefficient was used to obtain test-retest reliability. RESULTS: Three of the five evaluated programs offered a semi-automated technique to segment the images based on histogram values or a user-defined threshold. One software package allowed manual delineation only. One fully automated program demonstrated the drawbacks of uncritical automated processing. The semi-automated approaches reduced variability and measurement error, and improved reproducibility. There was no significant difference in the intra-observer agreement in SAT and CSA. The VAT measurements showed significantly lower test-retest reliability. There were some differences between the software packages in qualitative aspects, such as user friendliness. CONCLUSION: Four out of five packages provided essentially the same results with respect to the inter- and intra-rater reproducibility. Our results using SliceOmatic, Analyze or NIHImage were comparable and could be used interchangeably. Newly developed fully automated approaches should be compared to one of the examined software packages.

Can proton MR spectroscopic and perfusion imaging differentiate between neoplastic and nonneoplastic brain lesions in adults?

BACKGROUND AND PURPOSE: Noninvasive diagnosis of brain lesions is important for the correct choice of treatment. Our aims were to investigate whether 1) proton MR spectroscopic imaging ((1)H-MRSI) can aid in differentiating between tumors and nonneoplastic brain lesions, and 2) perfusion MR imaging can improve the classification. MATERIALS AND METHODS: We retrospectively examined 69 adults with untreated primary brain lesions (brain tumors, n = 36; benign lesions, n = 10; stroke, n = 4; demyelination, n = 10; and stable lesions not confirmed on pathologic examination, n = 9). MR imaging and (1)H-MRSI were performed at 1.5T before biopsy or treatment. Concentrations of N-acetylaspartate (NAA), creatine (Cr), and choline (Cho) in the lesion were expressed as metabolite ratios and were normalized to the contralateral hemisphere. Dynamic susceptibility contrast-enhanced perfusion MR imaging was performed in a subset of patients (n = 32); relative cerebral blood volume (rCBV) was evaluated. Discriminant function analysis was used to identify variables that can predict inclusion in the neoplastic or nonneoplastic lesion groups. Receiver operator characteristic (ROC) analysis was used to compare the discriminatory capability of (1)H-MRSI and perfusion MR imaging. RESULTS: The discriminant function analysis correctly classified 84.2% of original grouped cases (P < .0001), on the basis of NAA/Cho, Cho(norm), NAA(norm), and NAA/Cr ratios. MRSI and perfusion MR imaging had similar discriminatory capabilities in differentiating tumors from nonneoplastic lesions. With cutoff points of NAA/Cho < or =0.61 and rCBV > or =1.50 (corresponding to diagnosis of the tumors), a sensitivity of 72.2% and specificity of 91.7% in differentiating tumors from nonneoplastic lesions were achieved. CONCLUSION: These results suggest a promising role for (1)H-MRSI and perfusion MR imaging in the distinction between brain tumors and nonneoplastic lesions in adults.

Antioxidant supplementation reduces inter-individual variation in markers of oxidative damage.

The aim of this study was to examine the effect of antioxidant supplementation on oxidative damage and chromosome stability in middle-aged men, smokers and non-smokers. A total of 124 men aged 48+/-6 years from Bratislava and from the rural population near Bratislava were investigated; 64 men (22 smokers and 42 non-smokers) were supplemented for 12 weeks with antioxidants, while 60 (25 smokers and 35 non-smokers) were given placebo. The daily antioxidant supplementation consisted of vitamin C (100 mg), vitamin E (100 mg), ss-carotene (6 mg), and selenium (50 microg). Samples of blood were taken on two occasions: At the beginning and at the end of the supplementation trial. Concentrations of dietary antioxidants, ferric reducing ability, malondialdehyde as an indicator of lipid peroxidation in plasma, micronuclei and chromosome aberrations in lymphocytes were measured. Antioxidant supplementation significantly increased the levels of vitamin C, ss-carotene, a-tocopherol and selenium in plasma. The overall antioxidant status of plasma measured as ferric reducing ability of plasma (FRAP) increased significantly (p<0.001) after antioxidant supplementation as well. The increase in antioxidant parameters after supplementation were consistently more pronounced in non-smokers than in smokers. There was a significant decrease of malondialdehyde concentration in the non-smokers, while in smokers the decrease of malondialdehyde concentration was not significant. Antioxidant supplementation did not affect the proportion of lymphocytes with micronuclei or the total number of micronuclei; however, there was a significant positive correlation (p<0.001) between the malondialdehyde concentration at the beginning of the supplementation trial and the difference in number of cells with micronuclei before and after the supplementation. The percent of cells with chromosome aberrations decreased significantly after antioxidant supplementation in smokers. These results indicate that a combined antioxidant supplementation (a) is effective even at very moderate doses; (b) significantly diminishes oxidative damage to lipids when it is high initially; and (c) is effective in decreasing chromosomal instability in lymphocytes of middle-aged men.

Lipid peroxidation in men after dietary supplementation with a mixture of antioxidant nutrients.

Antioxidants and antioxidant enzymes protect living organisms against the attack of reactive oxygen species. An adequate daily intake of the individual antioxidants is therefore important to prevent the cells against oxidative damage. We investigated the effect of a modest dietary supplementation with a mixture of antioxidant nutrients (100 mg vitamin E, 100 mg vitamin C,6 mg beta-carotene and 50 microg of selenium per day) for 3 months on the plasma antioxidant capacity and indices of oxidative stress. Two groups of middle-age men were selected: group 1 with survivors of myocardial infarction (MI), and group 2 with clinically normal controls. The values of total antioxidant capacity of plasma (FRAP) significantly increased after supplementation with antioxidants in the both groups. Markers of in vivo lipid peroxidation, plasma malondialdehyde (MDA) and conjugated diene (CD) levels significantly decreased in the both supplemented groups. MDA and CD values were significantly higher at baseline in the group of survivors of myocardial infarction when compared with the group of healthy men. The results demonstrate that short-term and modest supplementation with a mixture of antioxidant nutrients improves antioxidative capacity and reduces products of lipid peroxidation in plasma. Since a more pronounced effect was observed within the group of survivors of myocardial infarction, a recommendation of antioxidant supplements seems appropriate for patients with a history of cardiovascular disease. (Fig. 1, Ref. 19.)

Folate levels determine effect of antioxidant supplementation on micronuclei in subjects with cardiovascular risk.

We have investigated the effect of modest supplementation with alpha-tocopherol (100 mg/day), beta-carotene (6 mg/day), vitamin C (100 mg/day) and selenium (50 microg/day) on oxidative stress and chromosomal damage, and the influence of methylenetetrahydrofolate reductase (MTHFR) genotype on these end-points. Subjects were two groups of middle-aged men differing in cardiovascular risk; 46 survivors of myocardial infarction before age 50 and 60 healthy controls. They were randomly divided into equal groups to receive antioxidants or placebo for 12 weeks. Twenty-eight patients and 58 controls completed the intervention. Micronucleus levels in peripheral lymphocytes and changes seen after intervention were studied in relation to the MTHFR C677T genotype, basal homocysteine and plasma folate levels. Ferric reducing ability of plasma and concentration of malondialdehyde were measured to assess the antioxidant effect of supplementation. There was no association of micronuclei with folate, homocysteine or malondialdehyde levels before supplementation. Micronucleus frequencies and plasma folate levels did not vary significantly with MTHFR genotype. Homocysteine levels in subjects with the TT variant genotype were significantly higher compared with CT or CC (P = 0.001), especially in subjects with low folate (P = 0.012). In the placebo control group an increase in micronuclei (P = 0.04) was detected at the end of the intervention period. This effect was not seen in the supplemented group. In antioxidant-supplemented myocardial infarction survivors we found an increase in the ferric reducing ability of plasma (P < 0.001) and a decrease in malondialdehyde (P = 0.001). Micronucleus frequency showed a decrease, strongest in subjects with normal folate levels (P = 0.015). In subjects with low folate levels, a high correlation was found between micronuclei after supplementation and homocysteine, both before (r = 0.979, P = 0.002) and after supplementation (r = 0.922, P = 0.009). Thus, folate deficiency may amplify the effect of other risk factors such as elevated homocysteine levels or variant MTHFR genotype, as well as influencing the ability of antioxidant supplementation to protect against genetic damage.

Leukoencephalopathy, cerebral calcifications, and cysts: new observations.

We describe three cases of the rare syndrome of leukoencephalopathy, brain calcifications, and cysts. Conventional MRI, proton spectroscopy, and diffusion-weighted imaging yielded additional information on the disease. Imaging findings favor increased water content rather than a demyelinating process in the pathophysiology of this disease. Clinical features of Coats disease and consanguinity were also encountered.

Nutritional supplementation with antioxidants decreases chromosomal damage in humans.

In order to investigate the effects of antioxidant supplementation on chromosome damage, a 3 month antioxidant supplementation trial was conducted on groups of 28 myocardial infarction survivors and 57 rural controls, all male. The supplement consisted of vitamin C (100 mg/day), vitamin E (100 mg/day), beta-carotene (6 mg/day) and selenium (50 microg/day). Dietary antioxidants in plasma were measured, as well as the ferric reducing ability of plasma (a measure of total plasma antioxidant status) and the concentration of malondialdehyde as an indicator of oxidative stress. Lymphocytes collected at the beginning and end of the supplementation period were stimulated to proliferate and metaphases accumulated for scoring of chromosome aberrations: per cent aberrant cells and chromatid and chromosome breaks. Supplementation with antioxidants was associated with a decrease in the percentage of cells with chromosome aberrations in the group of rural controls (0.63% before compared with 0.27% after supplementation; P = 0.03). The largest effect of supplementation was seen in smokers in this group (0.12% aberrant cells in supplemented compared with 0.81% in placebo group; P > 0.001). The results support the hypothesis that antioxidants decrease genetic damage.

Rapid method for correction of CSF partial volume in quantitative proton MR spectroscopic imaging.

Partial volume effects with cerebrospinal fluid (CSF), if uncorrected, can lead to underestimation of metabolite concentrations in quantitative proton magnetic resonance spectroscopic imaging (MRSI) of the brain. A rapid method for the correction of CSF partial volume effects is described based on selective CSF imaging using long echo time (TE) fast spin echo (FSE) magnetic resonance imaging (MRI). In order to achieve maximum suppression of signal from brain parenchyma, the FSE sequence is coupled with an inversion recovery (IR) pulse. Scan time is minimized using single shot (SS) IR-FSE. The method is validated against a current "gold standard" for the determination of CSF volumes, namely, segmented 3D spoiled gradient-echo (SPGR) scans. Excellent agreement in CSF percentage determined by the two methods was found (linear regression analysis: slope = 0.99 +/- 0.02, intercept = 2.08 +/- 0.45; mean +/- standard errors, R = 0.93) in pooled data from four healthy subjects. An example of the use of SS-IR-FSE for partial volume correction in a leukodystrophy patient with T(2) hyperintense lesions is demonstrated. SS-IR-FSE is a simple and rapid method for applying partial volume corrections in quantitative proton MRSI, which may be of particular value in the clinical environment when time constraints do not allow longer, perhaps more accurate segmentation methods to be used.

Neuroimaging studies in Rett syndrome.

Neuroimaging is a key instrument for determining structural and in vivo functional status of the brain, non-invasively. Multiple approaches can now determine aspects of anatomic and neurochemical changes in brain, and have been utilized effectively in Rett Syndrome patients to understand the biological basis of this neurodevelopmental disorder. Studies performed at our institute include volumetric analyses of MRI, magnetic resonance spectroscopy (MRS), diffusion tensor imaging (DTI), cerebral blood flow measurements with MRI, and positron emission tomography scans (PET). These studies have provided considerable insight into mechanisms underlying the clinical features of this disease. Volumetric analyses suggest that decreased brain volume in RS results from global reductions in both gray and white matter of the brain. A selective vulnerability of the frontal lobes is evidenced by the preferential reduction of blood flow, increased choline and reduced n-acetyl aspartate (NAA) by MRS, and increased glucose uptake in these same regions as shown by ((18)F)-fluorodeoxyglucose (FDG) PET scans. We hypothesize that the increased glucose uptake relates to increased glutamate cycling in synapses. The resulting neuroexcitotoxic injury to the developing brain contributes to the seizures, behavioral disturbance and respiratory irregularities commonly seen in phases 1 and 2 of this disorder.

Quantitative proton MR spectroscopic imaging of normal human cerebellum and brain stem.

Quantitative, multislice proton MR spectroscopic imaging (MRSI) was used to investigate regional metabolite levels and ratios in the normal adult human posterior fossa. Six normal volunteers (36 +/- 3 years, five male, one female) were scanned on a 1.5 T scanner using multislice MRSI at long echo time (TE 280 msec). The entire cerebellum was covered using three oblique-axial slice locations, which also included the pons, mid-brain, insular cortex, and parieto-occipital lobe. Concentrations of N-acetylaspartate (NAA), choline (Cho), and creatine (Cr) were estimated using the phantom replacement technique. Regional variations of the concentrations were assessed using ANOVA (P < 0.05). High-resolution MRSI data was obtained in all subjects and brain regions examined. Metabolite concentrations (mM) (mean +/- SD) were as follows: cerebellar vermis: 2.3 +/- 0.4, 8.8 +/- 1.7 and 7.6 +/- 1.0 for Cho, Cr, and NAA respectively; cerebellar hemisphere: 2.2 +/- 0.6, 8.9 +/- 2.1, 7.5 +/- 0.8; pons 2.2 +/- 0.5, 4.3 +/- 1.1, 8.3 +/- 0.9; insular cortex, 1.8 +/- 0.5, 7.8 +/- 2, 8.0 +/- 1.1, parieto-occipital gray matter, 1.3 +/- 0.3, 5.7 +/- 1.1, 7.2 +/- 0.9, and occipital white matter, 1.4 +/- 0.3, 5.3 +/- 1.3, 7.5 +/- 0.8. Consistent with previous reports, significantly higher levels of Cr were found in the cerebellum compared to parieto-occipital gray and occipital white matter, and pons (P < 0.0001). NAA was essentially uniformly distributed within the regions chosen for analysis, with the highest level in the pons (P < 0.04). Cho was significantly higher in the cerebellum and pons than parieto-occipital gray and occipital white matter (P < 0.002) and was also higher in the pons than in the insular cortex (P < 0.05). Quantitative multislice MRSI of the posterior fossa is feasible and significant regional differences in metabolite concentrations were found.

Kiwifruit protects against oxidative DNA damage in human cells and in vitro.

Antioxidant micronutrients may account for the beneficial effects of fruits on human health. A direct demonstration that consumption of fruit decreases oxidative DNA damage in human cells would support this hypothesis. Kiwifruit was taken as an example of a food with putative antioxidant properties, and its effectiveness at decreasing oxidative DNA damage was assessed in ex vivo as well as in vitro tests. The comet assay (single-cell gel electrophoresis) was used to measure DNA damage in lymphocytes collected during a human supplementation trial with a single 0.5-liter drink of kiwifruit juice (with water as a control). The comet assay was also modified to assess the antioxidant effect of kiwifruit in vitro by measuring the ability of an extract to interfere with oxidative damage to DNA induced by H2O2. Ex vivo, consumption of kiwifruit led to an increased resistance of DNA to oxidative damage induced by H2O2 in isolated lymphocytes, in comparison with lymphocytes collected after a control drink of water. No effect was seen on endogenous DNA damage. In vitro, a simple extract of kiwifruit, buffered to pH 7, was more effective than a solution of vitamin C (of equivalent concentration) at protecting DNA from damage, whereas at the highest concentrations tested, neither kiwi extract nor vitamin C had a protective effect. We have demonstrated significant antioxidant activity of kiwifruit ex vivo and in vitro, not attributable entirely to the vitamin C content of the fruit. Our dual approach is appropriate for testing other fruit and vegetable products for potential antioxidant effects.

Glutathione S-transferase polymorphisms influence the level of oxidative DNA damage and antioxidant protection in humans.

Glutathione S-transferase genotypes GSTT1, GSTM1, GSTP1 were characterised in 155 middle-aged men and compared with parameters of oxidative stress at the level of DNA and lipids, with antioxidant enzymes, and with plasma antioxidants in smokers and non-smokers. Smokers had on average significantly lower levels of Vitamin C, beta-carotene and beta-cryptoxanthin and higher amounts of oxidised purines and pyrimidines in lymphocyte DNA. The GSTM1 null genotype was associated with elevated glutathione as well as with higher Vitamin C concentration in plasma. Vitamin C was higher in GSTT1+ compared with GSTT1 null--as was glucose-6-phosphate dehydrogenase activity. The homozygous GSTP1 a/a genotype was associated with significantly higher levels of GST activity measured in lymphocytes, in comparison with the b/b genotype. Using multifactorial statistical analysis we found significant associations between smoking, GSTP1 genotype, plasma Vitamin C, and purine base damage in lymphocyte DNA. The difference in Vitamin C plasma levels between smokers and non-smokers was seen only with the GSTP1 b/b genotype. This group accounted also for most of the increase in purine oxidation in smokers. In contrast, the link between smoking and oxidised pyrimidines in DNA was seen only in the GSTT1 null group. It seems that polymorphisms in the phase II metabolising enzyme glutathione S-transferase may be important determinants of commonly measured biomarkers.

Proton magnetic resonance spectroscopy of choroid plexus tumors in children.

A variety of lesions may present as intraventricular masses in children. We report quantitative proton magnetic resonance spectroscopy (MRS) of two intraventricular tumors of the choroid plexus: choroid plexus carcinoma (CPC) and choroid plexus papilloma (CPP). Both lesions were characterized by high levels of choline-containing compounds and a complete absence of creatine and the neuronal/axonal marker N-acetyl aspartate. The CPC showed higher levels of choline compared to the CPP, and it also had elevated lactate. These preliminary results, if confirmed in a larger cohort of patients, indicate that proton MRS may have a role in the presurgical diagnosis of choroid plexus tumors in children, which may also have important implications for therapy and prognosis.

Detection of alkylation damage in human lymphocyte DNA with the comet assay.

The enzyme 3-methyladenine DNA glycosylase II (AlkA) is a bacterial repair enzyme that acts preferentially at 3-methyladenine residues in DNA, releasing the damaged base. The resulting baseless sugars are alkali-labile, and under the conditions of the alkaline comet assay (single cell gel electrophoresis) they appear as DNA strand breaks. AlkA is no t lesion-specific, but has a low activity even w ith undamagedbases. We have tested the enzyme at different concentrations to find conditions that maximise detection of alkylated bases with minimal attack on normal, undamaged DNA. AlkA detects damage in the DNA of cells treated with low concentrations of methyl methanesulphonate. We also find low background levels of alkylated bases in normal human lymphocytes.

The relationship between creatine kinase kinetics and exercise intensity in human forearm is unchanged by age.

Using (31)P magnetic resonance spectroscopy, creatine kinase (CK) reaction kinetics was assessed in the forearm flexor digitorum profundus muscle of healthy young (n = 11, age 34.7 +/- 5 yr) and older (n = 20, age 73.5 +/- 8 yr) subjects at rest, intermittent exercise at 20% maximum voluntary contraction (MVC), and 40% MVC. Exercise resulted in a significant increase in the average ratio of inorganic phosphate (P(i)) to phosphocreatine (PCr) from resting values of 0.073 +/- 0.031 (young) and 0.082 +/- 0.037 (older) to 0. 268 +/- 0.140 (young, P < 0.01) and 0.452 +/- 0.387 (older, P < 0. 01) at 40% MVC. At 40% MVC, intracellular pH decreased significantly, from resting values of 7.08 +/- 0.08 (young) and 7.08 +/- 0.11 (older) to 6.84 +/- 0.19 (young, P < 0.05) and to 6.75 +/- 0.25 (older, P < 0.05). Average values of the pseudo-first-order reaction rate k((PCr-->ATP)) at rest were 0.07 +/- 0.04 s(-1) in the young and 0.07 +/- 0.03 s(-1) in the older group. At both exercise levels, the reaction rate constant increased compared with the resting value, but only the difference between the resting value and the 20% MVC value, which showed an 86% higher reaction rate constant in both groups, reached statistical significance (P < 0.05). No difference in the reaction rate constant between the young and older groups was observed at either exercise level. As with k((PCr-->ATP)), the average phosphorus flux through the CK reaction increased during exercise at 20% MVC (P < 0.05 in the older group) but decreased toward resting values at 40% MVC in both groups. The data in our study suggest that normal aging does not significantly affect the metabolic processes associated with the CK reaction.

Metabolic heterogeneity at the level of the anterior and posterior commissures.

Multislice, 2D proton spectroscopic imaging was performed in six healthy volunteers at long echo time (TE = 280 msec). The center of the most inferior of three slices was placed directly at the level of the line connecting the anterior and posterior commissures. Significant regional variations in metabolite levels were observed. In particular, based on statistical analysis, levels of choline were significantly high in insular cortex, thalamus, and centrum semiovale compared to other brain regions such as parietal or occipital gray and white matter. NAA levels were highest in the centrum semiovale white matter, while creatine levels were relatively constant. Globus pallidus exhibited lower signal intensities and increased linewidths for all metabolites. No spectra could be obtained from the inferior frontal lobe because of field inhomogeneity. These data show that the metabolism, and perhaps the underlying cellular composition, of thalamus and insular cortex appears to be different from other neocortical gray matter. Normal regional variations in the brain spectra should be considered when evaluating pathological conditions.