Effect of quantity and quality of dietary protein on choline acetyltransferase and nerve growth factor, and their mRNAs in the cerebral cortex and hippocampus of rats.

The brain protein synthesis is sensitive to the dietary protein; however, the role of dietary protein on biomarkers including choline acetyltransferase and nerve growth factor (NGF) for the function of cholinergic neurons remains unknown in young rats. The purpose of this study was to determine whether the quantity and quality of dietary protein affects the concentration of NGF and activity of choline acetyltransferase, and their mRNA levels in the brains of young rats. Experiments were carried out on five groups of young rats (4 weeks) given the diets containing 0, 5, 20% casein, 20% gluten or 20% gelatin for 10 days. The activity of choline acetyltransferase in the cerebral cortex and hippocampus declined gradually with a decrease in quantity and quality of dietary protein. The concentration of NGF in the cerebral cortex and the mRNA levels of choline acetyltransferase in the cerebral cortex and hippocampus did not differ among groups. However, the concentration and mRNA level of NGF in the hippocampus was significantly lower in rats fed with lower quantity of protein or lower quality of protein. In the hippocampus, the mRNA levels of NGF significantly correlated with the NGF concentration when the quantity (r = 0.704, P < 0.01) and quality (r = 0.682, P < 0.01) of dietary protein was manipulated. It was further found that a significant positive correlation existed between the NGF concentration and the activity of choline acetyltransferase in the hippocampus (dietary protein quantity, r = 0.632, P < 0.05; dietary protein quality, r = 0.623, P < 0.05). These results suggest that the ingestion of lower quantity and quality of dietary protein are likely to control the mRNA level and concentration of NGF, and cause a decline in the activity of choline acetyltransferase in the brains of young rats.

Assessment of postischemic reperfusion and diamox activation test in stroke using 99mTc-ECD SPECT.

To evaluate the cerebral distribution of 99mTc-ethyl cysteinate dimer (99mTc-ECD) at blood flow levels beyond the normal range, we investigated postischemic reperfusion and acetazolamide (Diamox) activation test in stroke patients. The postischemic reperfusion was studied in 10 patients who showed a postischemic hyperperfusion area on other single photon emission computed tomography (SPECT) studies using N-isopropyl-rho-[123I]iodoamphetamine ([123I]IMP), 99mTc-hexamethyl propyleneamine oxime (99mTc-HMPAO), or 133Xe. 99mTc-ECD SPECT demonstrated a hyperactive area in one case, an isoactive area in four, and a hypoactive area in five. Correlations with CT findings revealed hyperactive areas without any abnormality, isoactive areas with perifocal rim, perifocal edema, or diffuse cerebral edema, and hypoactive areas with an infarct core. The Diamox activation test was studied in eight other patients with atherothrombotic stroke, and a limitation in vasodilative capacity was classified into three grades: Gr. 0 (none to minimal), Gr. I (mild), and Gr. II (moderate). [123I]IMP SPECT showed Gr. II and limitation in all eight cases. However, 99mTc-ECD showed Gr. II in three cases and Gr. I in five, and 99mTc-HMPAO revealed Gr. II in two cases, Gr. I in three, and Gr. 0 in three. We suggest that a lack of retention of 99mTc-ECD in a postischemic reperfusion area indicates the severity of the initial brain damage. Although the limitation in vasodilative capacity under Diamox-activated conditions was underestimated using 99mTc-labeled CBF tracers as compared with [123I]IMP, a retention of 99mTc-ECD in the unaffected area with an increased CBF under Diamox activation could be relatively superior to 99mTc-HMPAO.

Cerebral protein kinase C and its mRNA level in apolipoprotein E-deficient mice.

It is known that protein kinase C (PKC) activity may be one of the fundamental cellular changes associated with memory function. Apolipoprotein E (apoE) deficiency causes cholinergic deficits and memory impairment. ApoE-deficient mouse has been employed as a serviceable model for studying the relation between apoE and the memory deficit induced by cholinergic impairment. Brain-fatty acid binding protein (b-FABP) might be functional during development of the nervous system. Peroxisome proliferator-activated receptor (PPAR) is involved in the early change in lipid metabolism. We investigated the alterations not only in cerebral PKC activity, but also in the gene expressions of PKC-beta, brain-FABP and PPAR-alpha in apoE-deficient mice. The results showed that there was a lower cerebral membrane-bound PKC activity in the apoE-deficient mice than in its wild type strain (C57BL/6). But there were no significant differences in cytosolic PKC activity. PKC-beta, b-FABP and PPAR-alpha mRNA expressions in cerebrum were lowered in apoE-deficient mice. These findings may be involved in the dysfunction of the brain neurotransmission system in apoE-deficient mouse. Alternatively, these results also suggest that cerebral apoE plays an important role in brain PKC activation by maintaining an appropriate expression of b-FABP and PPAR-alpha mRNAs.

Mechanism of the desmutagenic effect of humic acid.

The mechanism of an apparent desmutagenic effect of humic acid was investigated. Firstly, components of humic acid (resorcinol, vanillin, vanillic acid, ferulic acid, protochatechuic acid and benzoic acid) were tested and were not found to show a desmutagenic effect. By contrast, lignin did show a desmutagenic effect. The desmutagenic effect of humic acid was decreased by ozone treatment, and the degree of decrease corresponded with a decrease in KMnO4 consumption. Benzo[a]pyrene and humic acid were incubated at 37 degrees C for 1 h and extracted by ethyl acetate and the extract was investigated by gas chromatography (GC). The peak of the decomposition product did not appear, but the amount of benzo[a]pyrene was decreased. This suggests that the desmutagenic effect of humic acid was caused by adsorption of benzo[a]pyrene by humic acid rather than by decomposition of benzo[a]pyrene. Humic acid had the largest adsorption activity at its critical micelle concentration (CMC), while adsorbed benzo[a]pyrene could be released by ultrasonication. Fulvic acid and water-soluble humic substance showed a slight inhibitory effect on the mutagenicity of benzo[a]pyrene.

Determination of organically-associated trace metals in estuarine sea-water by solvent extraction and atomic-absorption spectrometry.

Chloroform extraction of trace metals (Ni, Cu, Mo, Mn, Cd and Pb) in estuarine sea-water was studied at pH 8 and pH 3, on the basis that the metals would be associated with dissolved organic matter (DOM), which has recently been characterized by reversed-phase liquid chromatography. Ni, Cu, Mo and Mn were extracted more at pH 8 than at pH 3. Cd and Pb were not associated with the DOM at either pH 8 or 3. The percentage of the total dissolved trace metals in sea-water associated with DOM varied from 0 to 14%. The metals extracted into chloroform at pH 8 were assumed to be associated with neutral or weakly basic DOM while at pH 3 they could be associated with either the neutral (or weakly basic) DOM or two types of acidic DOM.

Structural alterations of the liver induced by N,N'-diacetylbenzidine. A new model of cirrhosis.

Little is known about hepatic structural alterations induced by administration of the aromatic amine, N,N'-diacetylbenzidine. Therefore, we injected adult rats with this chemical at a dose of approximately 1 g/kg of body weight. Light and electron microscopy of liver specimens revealed hepatocellular degeneration and prominent changes in bile canaliculi, including dilation, disappearance of microvilli, and an apparent loss of structural integrity of the cell membrane. These findings were seen in the peripheral part of the hepatic lobules. Cirrhotic changes eventually developed. We concluded that N,N'-diacetylbenzidine injected intraperitoneally is capable of inflicting structural damage to the liver in rats.

Effect of suspension hypokinesia/hypodynamia on tissue protein turnover in rats.

The effect of suspension hypokinesia/hypodynamia on the protein metabolism in skeletal muscles (gastrocnemius and soleus) and visceral organs (liver and small intestine) was investigated in the rat. Suspension for 10 days resulted in atrophy of the visceral organs as well as the skeletal muscles, which was associated with decreases in the amounts of visceral and muscle protein. There was marked breakdown of muscle protein, which was reflected by increases in the urinary excretion of N7-methylhistidine and cathepsin D activity. Measurement of protein synthesis by the L-[4-3H]phenylalanine method revealed that the synthesis in the gastrocnemius, but not in the soleus, muscle was suppressed on suspension for 10 days. Thus, both the increased catabolism and decreased synthesis of protein in the muscles may be causally related to the muscle atrophy occurring in suspension. In the visceral organs, on the other hand, the protein synthesis was found to increase in hypokinetic rats. Moreover, the concentration of visceral protein was also increased, despite the decreased amount of muscle protein. These changes in protein metabolism in the liver and small intestine may explain, at least partly, the slightly positive nitrogen balance which was observed after long-term weightlessness.

Adsorption of mutagens by humic acid.

Humic acid inhibited the mutagenicity of various mutagens. The inhibitory effect was desmutagenic, heat-resistant and increased with an increase of the humic acid molecular weight. Typical monomeric components of humic acids had no desmutagenic effect. The desmutagenic effect of humic acid was caused by adsorption of mutagen, not by decomposition of mutagen. The adsorption activity was largest at its critical micelle concentration and the adsorbed mutagen was released by ultrasonication. Humic acids exist in natural environment in large amounts and may play an important role for natural purification by adsorption of mutagens.

Role of ornithine transport into mitochondria in urea synthesis of rats treated in thyroid hormone.

The purpose of this study was to find whether or not the ornithine transport into mitochondria regulated urea synthesis when the thyroid status is manipulated. Experiments were done on three groups of rats: given 6-propyl-2-thiouracil (PTU, a thyroid inhibitor) without triiodothyronine (T3) treatment, treated with PTU+T3 or receiving neither PTU nor T3 (control). The urinary excretion of urea, liver concentration of ornithine and ornithine transport into isolated hepatic mitochondria in rats given PTU+T3 were significantly lower than in rats given PTU alone. Ornithine transport was significantly inhibited by the addition of lysine specifically. This response was achieved well within the physiological concentration of lysine. Compared with rats given PTU without T3 treatment, the liver concentration of lysine was significantly higher in rats treated with PTU+T3 and control rats. Ornithine transport into hepatic mitochondria was closely correlated with the excretion of urea. The results suggest that the greater ornithine transport in the hypothyroid (PTU alone) rats is likely to stimulate urea synthesis. A thyroid hormone-induced increase in lysine concentration may be at least partly responsible for the changes in ornithine transport into mitochondria.

Effect of dietary protein quality on the brain protein synthesis rate in aged rats.

The purpose of this study was to determine whether the quality of dietary protein affects the rate of brain protein synthesis in aged rats. Experiments were conducted on three groups of aged rats (30 wk) given diets containing 20 g casein, 20 g gluten or 20 g gelatin/100 g for 10 d. The fractional rates of protein synthesis in the brain, liver and kidney declined with the decrease in quality of dietary protein. In the brain, liver and kidney, the RNA activity [g protein synthesized/(g RNA.d)] correlated significantly with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that the rate of protein synthesis in the brain declines with the decrease in quality of dietary protein consumed by aged rats, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis.

Effects of dietary protein restriction on the fractional rates of protein synthesis in perfused rat hindlimb.

The effects of dietary protein restriction on protein synthesis were investigated in perfused rat hindlimb. The fractional rate of protein synthesis was measured with [3H]phenylalanine in young adult (7-week-old) rats fed a low protein (5% casein) diet and a protein-free diet for 3 weeks. The low protein diet (LPD) allowed a moderate gain in body weight. The fractional rate of protein synthesis fell to 70% of the control value in LPD group and further fell to less than a half in the protein-free diet (PFD) group. Thus, the protein synthesis rate decreased as the dietary protein content was reduced. The fall of protein synthesis was mainly accompanied by the reduction of RNA activity (mg protein/mg RNA/day) rather than RNA concentration (RNA/protein). The rate of protein breakdown was calculated by subtracting growth rate from protein synthesis rate. The breakdown rate was decreased in LPD group and increased slightly in PFD group. From the low rates of protein synthesis and breakdown, it appears that dietary protein restriction, at least allowing a gain in body weight, makes the turnover rate slow down. The overall changes in protein synthesis obtained in the perfused hindlimb are consistent with the reported results in vivo.

The role of growth hormone and amino acids on brain protein synthesis in aged rats given proteins of different quantity and quality.

The purpose of the present study was to determine whether the regulation of brain protein synthesis was mediated through changes in the plasma concentrations of insulin and growth hormone (GH), and whether the concentrations of amino acids in the brain and plasma regulate the brain protein synthesis when the quantity and quality of dietary protein is manipulated. Two experiments were done on three groups of aged rats given diets containing 20% casein, 5% casein or 0% casein (Experiment 1), and 20% casein, 20% gluten, or 20% gelatin (Experiment 2) for 1 d (only one 5-h period) after all rats were fed the 20% casein diet for 10 d (only 5-h feeding per day). The aggregation of brain ribosomes, the concentration in plasma GH, and the branched chain amino acids in the plasma and cerebral cortex declined with a decrease of quantity and quality of dietary protein. The concentration of plasma insulin did not differ among groups. The results suggest that the ingestion of a higher quantity and quality of dietary protein increases the concentrations of GH and several amino acids in aged rats, and that the concentrations of GH and amino acids are at least partly related to the mechanism by which the dietary protein affects brain protein synthesis in aged rats.

Dietary gamma-aminobutyric acid affects the brain protein synthesis rate in young rats.

The purpose of this study was to determine whether the gamma-aminobutyric acid (GABA) affects the rate of brain protein synthesis in male rats. Two experiments were done on five or three groups of young rats (5 wk) given the diets containing 20% casein administrated 0 mg, 25 mg, 50 mg, 100 mg or 200 mg/100 g body weight GABA dissolved in saline by oral gavage for 1 day (d) (Experiment 1), and given the diets contained 0%, 0.25% or 0.5% GABA added to the 20% casein diet (Experiment 2) for 10 d. The plasma concentration of growth hormone (GH) was the highest in rats administrated 50 mg and 100 mg/100 g body weight GABA. The concentration of serum GABA increased significantly with the supplementation groups. The fractional (Ks) rates of protein synthesis in brain regions, liver and gastrocnemius muscle increased significantly with the 20% casein + 0.25% GABA diet and still more 20% casein + 0.5% GABA compared with the 20% casein diet. In brain regions, liver and gastrocnemius muscle, the RNA activity [g protein synthesized/(g RNA . d)] significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. Our results suggest that the treatment of GABA to young male rats are likely to increase the concentrations of plasma GH and the rate of protein synthesis in the brain, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis.

Diagnosis of Vertebral Artery Dissection by Basi-parallel Anatomical Scanning (BPAS) MRI.

SUMMARY: To diagnose VA dissection, MRA or cerebral angiography, which provides information regarding intra-vascular space, has been performed. We report the acquisition of various information about VA dissection using MRI-BPAS, which is a new diagnostic method.

Cerebral Blood Flow Change Before and After Carotid Angioplasty and Stenting (CAS) in Cases with Contralateral Carotid Artery Occlusion.

SUMMARY: Contralateral carotid artery occlusion is thought to represent a significant risk factor in carotid endarterectomy (CEA). There is also evidence that intraoperative and postoperative hypotention may cause contralateral hemodynamic ischemia. As such, contralateral carotid artery occlusion is regarded as a risk factor for carotid angioplasty and stenting (CAS). In this paper, we report on five cases of severe ICA stenosis with contralateral carotid artery occlusions. Cerebral blood flow(CBF) and cerebral vasoreactivity( CVR) of the contralateral carotid artery occlusions were measured before and after CAS. Additionally, the influence that ipsilateral CAS exerted on the occluded side was examined. (123)I-IMP SPECT was performed before and after CAS, both at rest and at the time of acetazoramide administration. The CBF was evaluated quantitatively using the ARG method. The mean CBF of the treated side rose from 30.0 +/- 7.1 ml/100g/min to 34.4 +/- 8.3 ml/100g/min (p < 0.05), and the mean CBF of the occluded side similarly rose from 28.3 +/- 6.1 ml/100g/min to 31.7 +/- 6.4 ml/100g/min (p < 0.05). Correspondingly, the regional CVR (rCVR) increased from 5.9% +/- 16.3% to 35.0% +/- 16.4%(p < 0.05) on the treated side, and from 3.7% +/- 14.7% to 10.7% +/- 16.9% (p < 0.05) on the occluded side. This demonstrates that ipsilateral CAS seems to improve both CBF and CVR on the contralateral occluded side. The fact that some cases developed cross flow from the anterior communicating artery was both remarkable and significant. Where there was poor cross flow from the anterior communicating artery, improvement in cerebral vaso reactivity was limited.

Effect of adding dietary L-lysine, L-threonine and L-methionine to a low gluten diet on urea synthesis in rats.

We have shown that urinary urea excretion increased in rats fed a low quality protein. The purpose of present study was to determine whether an addition of dietary limiting amino acids affected urea synthesis in rats fed a low gluten diet. Experiments were done on three groups of rats given diets containing 10% gluten, 10% gluten +0.5% L-lysine or 10% gluten+0.5% L-lysine, 0.2% L-threonine and 0.2% L-methionine for 10 d. The urinary excretion of urea, and the liver concentrations of serine and ornithine decreased with the addition of dietary L-lysine, L-threonine and L-methionine. The fractional and absolute rates of protein synthesis in tissues increased with the treatment of limiting amino acids. The activities of hepatic urea-cycle enzymes was not related to the urea excretion. These results suggest that the addition of limiting amino acids for the low gluten diet controls the protein synthesis in tissues and hepatic ornithine and decline urea synthesis.