Kii ALS dementia.

Epidemiological surveys in the foci of ALS of the Kii Peninsula of Japan started in the early 1960s. Continuous surveys conducted for decades revealed that there have been two foci in the Kii Peninsula: one in Kozagawa in the southern part, and the other in Hobara in the south-east. Clinically, ALS patients of the Kii foci occasionally showed parkinsonian features or dementia that have not been reported in the sporadic form of ALS. Neuropathologically, numerous NFT that are identical to those of Alzheimer's disease were observed in the cerebral cortex and in the brainstem nuclei. To elucidate the etiopathogenesis of this unique form of ALS, an analysis was conducted of the environment in the focus areas and of the specimens from the patients with ALS. It was hypothesized that the long exposure of these environments to low calcium and magnesium, and an excess of aluminum and manganese in the drinking water and the soil, might lead to the deposition of some trace elements in the CNS, eventually causing neuronal degeneration and death.

Inhibitory effects of astragali radix, crude drug in Oriental medicines on lipid peroxidation and protein oxidative modification of mouse brain homogenate by copper.

Astragali Radix, the root of Astragalus membranaceus Bunge, is a crude drug used widely in Oriental medicines. It is a major component of Ougi-Keishi-gomotsu-to, a traditional herbal medicine, used for neurop patients with abnormal sensations and neuropathic pain of the legs. It was shown to have inhibitory effects on lipid peroxidation and protein oxidative modification by copper. The effects were similar to and stronger than those of mannitol and superoxide dismutase as free radical scavengers. These results demonstrated that Astragali Radix has inhibitory effects on oxidative stress induced by metal.

Inhibitory effects of Ougi-keishi-gomotsu-to, a traditional herbal medicine, on lipid peroxidation and protein oxidative modification of mouse brain homogenate induced by copper.

Ougi-Keishi-gomotsu-to a traditional herbal medicine, is used clinically for patients with abnormal sensations and pain in the legs in neuropathy. It was shown to have inhibitory effects on lipid peroxidation and protein oxidative modification of brain homogenate induced by copper. Its effect was stronger than those of mannitol and alpha-tocopherol as free radical scavenger and antioxidant. These results demonstrated that Ougi-kelshi-gomotsu-to has antioxidative effects on neuron injury derived from oxidative stress induced by metal.

Inhibition in vitro linoleic acid peroxidation and haemolysis by caffeoyltryptophan.

Antioxidant activities of caffeoyltryptophan were investigated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging system, the superoxide anion generation system and the superoxide anion-mediated linoleic acid peroxidation system. At 10 microM, caffeoyltryptophan showed greater scavenging activity on DPPH than dl-alpha-tocopherol or ascorbic acid. DPPH radical scavenging activity of caffeoyltryptophan increased dose-dependently at concentrations ranging from 1 to 50 microM; 1 mol of caffeoyltryptophan reacted with ca 4 mol of radical. Caffeoyltryptophan caused 80% inhibition of superoxide anion generation at 50 microM. The inhibitory activity of caffeoyltryptophan was as strong as that of 5-caffeoylquinic acid. Caffeoyltryptophan inhibited the formation of conjugated diene from linoleic acid. The inhibitory activity increased in the order caffeic acid < 5-caffeoylquinic acid < caffeoyltryptophan < dl-alpha-tocopherol. Effects on the in vitro haemolysis and peroxidation of mouse erythrocytes induced by H2O2 were also examined. Caffeoyltryptophan exhibited strong inhibitory activities; Tryptophan was ineffective in these systems. These data suggest that caffeoyltryptophan may be a natural antioxidant in the human diet and, as such, may intervene in toxicological processes that are mediated by radical mechanisms.

Epidemiology of motor neuron disease in the Kii Peninsula of Japan, 1989-1993: active or disappearing focus?

During the period 1989-1993, the incidence and migration patterns of patients with motor neuron diseases (MND) in Wakayama Prefecture, including one of the high-incidence Kii Peninsula foci ('Kozagawa focus'), were surveyed to determine whether the focus had truly disappeared or not. Overall, the crude average annual incidence was 1.43 per 100000 population; when age-adjusted to the 1990 Japanese population, it was 1.25 (1.85 for males and 0.61 for females). The average annual age- and sex-specific incidence steadily increased to a peak between 60 and 69 years and dropped after 70. Geographically, the rates varied in the five regions of Wakayama Prefecture from 0.38 to 2.48. The areas with high incidence were distributed in the central and southernmost regions; the highest was in the Kozagawa focus with 9.54 (two ALS cases within five years; 4193 base population, 1990). During the study period, four emigrants from Kozagawa had developed MND one to four decades after leaving the focus. Although the remarkable clustering of MND was thought to have disappeared, the southern Kii Peninsula remains a high-risk area for MND, especially if one interprets the data so as to include the emigrants. In general, the age at onset has increased in the past 20 years from 56.5 to 61.7; male predominance is observed.

Effect of aluminum on iron-induced lipid peroxidation and protein oxidative modification of mouse brain homogenate.

In the present study the authors report on the enhancing effect of aluminum(III) (Al[III]) on iron(II)(Fe[II])-induced lipid peroxidation (LPO) of mice brain homogenate, which occurs in a concentration- and time-dependent manner. No evidence of LPO caused by Al alone was found. Both Al(III) and Fe(II) ions induced protein oxidative modifications in mice brain homogenate, in a time- and concentration-dependent manner. Aluminum enhances Fe(II)-induced protein oxidative modification at a concentration of 2:1 and 1:1 Al:Fe molar ratios. However, Al suppress Fe(II)-induced protein oxidative modification at a concentration of 0.5:1 Al:Fe molar ratio. Addition of ethylenediaminetetraacetic acid (EDTA) inhibits both LPO and protein oxidative modifications induced by Al(III) and Fe(II) ions. Addition of mannitol and of superoxide dismutase (SOD) did not show such effects. It is concluded that in mice brain homogenate, Al accelerates Fe(II)-induced LPO. Protein oxidative modifications caused by Fe(II) and/or Al ions are enhanced at high, but suppressed at low concentrations of Al ions. The latter observation suggests a possible biological role of Al as an antioxidant.

Involvement of Onuf's nucleus in amyotrophic lateral sclerosis.

The Onufrowicz (Onuf's) nuclei from eight amyotrophic lateral sclerosis (ALS) cases and nine neurological control cases were studied histologically and morphometrically. To clarify the factors relating to the involvement of the Onuf's nucleus in ALS, we correlated the relationships among the age at death, clinical duration, morphometric findings for Onuf's neurons, and neuronal numbers in the posteroposterolateral (PPL) nuclei in the ALS cases with those in neurological controls. Intracytoplasmic inclusions such as Bunina bodies, ubiquitin-reactive inclusions, and conglomerate inclusion were found in the Onuf's neurons in ALS, but not in the controls. The total number of Onuf's neurons in the ALS cases was not decreased, but that of normal-appearing neurons was decreased while that of atrophic neurons was increased. Significantly decreased perikaryal, nuclear and nucleolar areas and decreased perikaryal (P)/nuclear and P/nucleolar area ratios of Onuf's neurons were found in ALS, not only in the atrophic neurons but also in the normal-appearing neurons, compared with the controls. The shrinkage in Onuf's neurons of ALS was different from that seen in the ageing process or in the axonal reactions of controls with atonic bladder. In ALS, the morphometric findings for the Onuf's neurons showed no correlation with age at death, clinical duration, or number of PPL neurons. Our results indicate that in ALS Onuf's nucleus is principally vulnerable to the ALS process, although the degree of degeneration differs from that seen in other motor neurons. The involvement of Onuf's nucleus might be slowed due to factors specific to this nucleus, including the biochemical and autonomic properties of the nucleus; nevertheless, it is histologically classified as part of the somatic cell column.

[Amyotrophic lateral sclerosis--causative role of trace elements].

Although numerous hypotheses have been proposed for the cause of amyotrophic lateral sclerosis (ALS), conclusive decision still remains vague. Recent epidemiological investigation disclosed an aggregation of ALS cases in the Western Pacific, including the Kii Peninsula of Japan, the island of Guam in Marianas and West New Guinea. Extensive environmental studies in these foci indicated an important role of trace elements in ALS etiology. It is postulated that chronic environment deficiencies of calcium and magnesium may provoke secondary hyperparathyroidism, resulting in increased intestinal absorption of toxic metals under the presence of excess levels of divalent or trivalent cations and lead to the mobilization of calcium and metals from the bone and deposition of these elements in nervous tissue. This hypothesis, called metal-induced calcifying degeneration of CNS, has been supported by experimental studies using several animal species.

Bunina body formation in amyotrophic lateral sclerosis: a morphometric-statistical and trace element study featuring aluminum.

Clinico-environmental and pathological variables were obtained from 10 patients with amyotrophic lateral sclerosis using particle-induced X-ray emission spectrometry (PIXE) and morphometric-statistical analysis. Statistical analysis identified a model that maximally predicts the Bb% (frequency of Bunina bodies) from a selected set, four variables: (1) nucleolar index, (2) magnesium (Mg) content, (3) aluminum (Al) content, and (4) duration of illness. Among them, only the Al content proved important. To determine their chemical nature, electron energy loss spectrometry (EELS) was applied at the ultrastructural level; it revealed that within the motor neuron, Al strongly binds to the Bunina body as well as rough endoplasmic reticulum (rER), and lesser strongly to mitochondria and lipofuscin granule. Thus, it is chemically similar to the rER, providing preferential binding sites to aluminum. The Bunina bodies may be an end-product of the nucleic acid dysmetabolism at rER caused by Al along with Mg depletion.

Aluminum-induced model of motor neuron degeneration: subperineurial injection of aluminum in rabbits.

Environmental factors, particularly chronic exposure to aluminum (Al) and manganese (Mn) with dietary deficiency of calcium (Ca) and magnesium (Mg), are speculated to be contributory in the pathogenesis of amyotrophic lateral sclerosis (ALS). However, the mechanisms by which these elements accumulate in the CNS tissues and induce neuronal death are not known. In the present study, we investigated the retrograde transport of Al as a possible mechanism of pathogenesis. Al (as aluminum chloride or maltol) was injected into the subepineurial space of the sciatic nerve with subsequent morphological evaluation of the neurotoxic effect on spinal motor neurons in rabbits. Spheroid/globules, central and peripheral chromatolysis, and neuronal degeneration were observed in the spinal anterior horn in Al-maltol, Al chloride, and maltol treated rabbits to more marked extent than those in uninjected or saline controls. By electron microscopy, the soma and dendrites of neurons in the anterior horn at the fifth lumbar spinal cord in the Al-treated rabbit showed marked edematous change, fragmentation of granular endoplasmic reticulum, increased accumulation of neurofilament, and accumulation of free ribosomes and lipid-droplet-like structures. Horseradish peroxidase (HRP) reactive product was seen in the axons and cytoplasm of Schwann cells of the sciatic nerve in Al-maltol treated rabbits, suggesting that the permeability of the blood-nerve-barrier was increased by injection of Al-maltol. We suggest that Al, subperineurially injected, was absorbed into the spinal cord and induced degeneration of spinal motor neurons in these rabbits. These findings indicate that the retrograde transport of Al into spinal motor neurons via the peripheral nervous system may exacerbate neuronal degeneration in ALS.

Lewy body-like inclusions in Onuf's nucleus from two cases of sporadic amyotrophic lateral sclerosis.

Lewy body-like inclusions in Onuf's neurons from two sporadic cases with amyotrophic lateral sclerosis (ALS) were reported. These inclusions in Onuf's neurons as well as those found in the anterior horn cells were immunostained with an anti-ubiquitin antibody. Neuropathological examination of these two cases revealed neuronal loss and associated gliosis in the anterior horn of the spinal cord and hypoglossal nuclei, and degeneration of the corticospinal tract. In addition to Lewy body-like inclusions, ubiquitinated skein-like inclusions, Bunina bodies, or both were observed in the cytoplasm of the remaining neurons in the anterior horn of the spinal cord, and, to a lesser degree, in Onuf's nucleus. Spheroids and cord-like thickening of cell processes were also found in the anterior horn of the spinal cord. Histometrical study of Onuf's nucleus revealed atrophy and loss of Onuf's neurons from Case 1 with a long clinical course. Similar cases of motor neuron disease with or without tract degeneration have been reported, but the presence of Lewy body-like inclusions in Onuf's nucleus is reported here for the first time. It is suggested that Onuf's nucleus is more or less involved in the degenerative process characteristic of ALS.

[The etiopathogenesis of amyotrophic lateral sclerosis--recent development].

In spite of extensive studies of amyotrophic lateral sclerosis (ALS) for the past decades, its cause still remains obscure. Based on the clinicopathological observations and experimental studies of sporadic and endemic ALS cases, etiologic hypothesis encompass a wide range of postulated pathophysiological mechanisms in the various fields. Reviewing briefly these, recent concern on genetic hypothesis, of familial ALS, background of case aggregation in the Western Pacific and immune hypothesis has been discussed. Although there is no unifying theory explanatory for ALS process at present, accumulated results through basic and clinical neurological science may provide a better understanding for the pathogenesis of ALS.

Magnesium and calcium contents in CNS tissues of amyotrophic lateral sclerosis patients from the Kii peninsula, Japan.

To further elucidate the involvement of metals as a factor in the pathogenesis of amyotrophic lateral sclerosis (ALS) on the Kii peninsula of Japan, a well-known high incidence area of ALS with low calcium and magnesium contents in soil and drinking water, we determined concentrations of these metals in samples of central nervous system tissue taken from postmortem ALS cases. Calcium content was determined by neutron activation analysis and magnesium by inductively coupled argon plasma emission spectrometry. From 5 ALS cases and 5 neurologically normal controls, we examined tissues from the precentral gyrus, including the motor cortex, internal capsule, crus cerebri and spinal cord, and from 22 other areas. The average calcium content in precentral gyrus, internal capsule, crus cerebri and spinal cord in ALS cases was higher than that in the controls, and the mean value of all 26 areas in the ALS cases was also higher than that of the controls. The average magnesium content of each region as well as the mean value of the 26 regions in the ALS cases was significantly lower than that in the controls. The Ca/Mg ratio of the 26 ALS regions was significantly higher than that of controls. This study strengthens our hypothesis that an abnormal metal metabolism plays a responsible role in the Ca-hydroxyapatite formation observed in central nervous system tissue of ALS cases, leading to motor neuron death and degeneration of the pyramidal tracts.

Distribution of calcium in central nervous system tissues and bones of rats maintained on calcium-deficient diets.

Current changing epidemiological pattern in the Western Pacific area suggests a contribution of the environmental factors to the pathogenetic process of amyotrophic lateral sclerosis (ALS). The condition of unbalanced mineral levels found in the soil and drinking water samples from the ALS foci showing low content of calcium (Ca) and magnesium (Mg) plus high content of aluminum (Al) was experimentally mimicked in this study using rats. In the groups fed low Ca, low Ca-Mg, and low Ca-Mg plus high Al diets, serum Ca levels were lower than that in the group fed the standard diet. Ca content of CNS tissues showed higher values in the unbalanced diet groups, especially in the spinal cord of low Ca-Mg plus high Al diet group, than those in the standard diet group, determined by inductively-coupled plasma emission spectrometry (ICP). Ca content in heart, liver, kidney, and abdominal aorta in groups fed low Ca-Mg, and low Ca-Mg plus high Al diets was higher than that in low Ca, and standard group. Ca content in muscle in the three unbalanced diet groups was significantly higher than in the standard diet group. Ca and Mg contents in lumbar spine and cortical bone showed lower values in the unbalanced diet groups than those values in the standard diet group. These findings suggest that under the condition of derangement of bone mineralization induced by unbalanced mineral diets fed to the experimental rats, Ca and Mg may be mobilized from bone, keeping their content in soft tissues, including CNS tissue, for utilization of vital activities, thereby resulting in a deposition of Ca while maintaining an almost normal value of magnesium in the CNS tissues.

Involvement of Onuf's nucleus in ALS. Demonstration of intraneuronal conglomerate inclusions and Bunina bodies.

Onuf's nucleus of 3 ALS cases was examined histologically, morphometrically and for metal content. Case 1 showed conglomerate inclusions (CIs), cases 2 and 3 showed Bunina bodies in Onuf's nucleus. Electron microscopy showed that CIs were intracytoplasmic accumulations of 10 nm neurofilaments with discrete borders. Onuf's neurons in ALS showed a significant decrease (P less than 0.001) in the cytoplasmic, nuclear and nucleolar areas in comparison with neurons of age-matched controls and elderly controls. Metal analysis of the sacral spinal cords by alpha particle-excited X-ray fluorescence analysis (PIXE) showed relatively high Al levels in the ALS cases compared with controls. Morin staining revealed intense green fluorescence (indicating Al) in the nucleoli and cytoplasm of the CI-containing neurons, but not in the CIs themselves. The appearance of CIs or Bunina bodies and neuronal atrophy in Onuf's nucleus seems to indicate that this structure is also involved in the disease process of ALS, although it is less vulnerable than most other motor ganglia.

[Evaluation of magnesium, calcium and aluminum deposition in bone in situ].

Current changing epidemiological pattern in the Western Pacific strongly suggests a contribution of the environmental factors to the pathogenetic process of amyotrophic lateral sclerosis (ALS). As a reflection of excess metals and a deficiency of minerals in soil and water samples in these foci, this study was designed experimentally to evaluate the concentration of calcium (Ca), magnesium (Mg) and aluminum (Al) in the bones of rats fed unbalanced mineral diets. Twenty-eight male Wistar rats, weighing 200 g, were fed either a standard diet, a low Ca diet, a low Ca-Mg diet, or a low Ca-Mg diet with high Al for 90 days. The composition of the diet/100 g consists of Ca 1250 mg, Mg 300 mg, Al 10 mg, Zn 4 mg in the standard diet; Ca 3 mg and Mg 2 mg in the low Ca-Mg diet; and Al 194 mg in the high Al diet, Al supplied as Al lactate. Ca, Mg and Al concentrations were determined by using inductively coupled plasma emission spectrometry (ICP) for bone and atomic absorption spectrometry for serum. Serum Ca levels in the groups fed unbalanced mineral diets were lower than those in the group fed standard diet. Serum Mg levels were markedly decreased in the groups fed low Ca-Mg diet and low Ca-Mg plus high Al diet, compared with those in the groups fed standard diet and low Ca diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Aluminum deposition in the central nervous system of patients with amyotrophic lateral sclerosis from the Kii Peninsula of Japan.

Chronic dietary deficiency of calcium (Ca) and magnesium (Mg) with excessive intake of aluminum (Al) and manganese (Mn) has been implicated in the pathogenesis of high incidence amyotrophic lateral sclerosis (ALS) in the Western Pacific. We report two cases of ALS from the Kii Peninsula of Japan with markedly elevated concentrations of Al in central nervous system (CNS) tissues. Six pathologically verified cases of ALS and five neurologically normal controls were studied. Levels of Al, Ca and phosphorus (P) were determined simultaneously by neutron activation analysis (NAA), and Mg concentration was measured by inductively coupled plasma emission spectrometry (ICP) in 26 CNS regions. Al concentrations in the precentral gyrus, internal capsule, crus cerebri and spinal cord were significantly increased in two ALS patients, compared with those of controls. Mean Al concentrations of the 26 CNS regions in these two patients were also higher than those of controls and of the four other ALS cases (p less than 0.01). By contrast, Mg concentrations in the 26 CNS regions were markedly reduced in the ALS cases, compared with controls (p less than 0.01), and the Ca/Mg ratios were significantly increased in the ALS cases (p less than 0.01). Our data indicate that high-incidence ALS in the Western Pacific may result from Ca-Mg dysmetabolism with resultant deposition of Al.

Evaluation of magnesium, calcium and aluminum metabolism in rats and monkeys maintained on calcium-deficient diets.

The epidemiology of amyotrophic lateral sclerosis (ALS) in the Western Pacific indicates that low concentrations of calcium (Ca) and magnesium (Mg) and high levels of aluminum (Al) in soil and water in these foci are etiologically important. To determine the biochemical derangements and metal deposition induced by chronic dietary deficiencies of Ca, we maintained experimental animals on several regimens. Male Wistar rats, weighing 100g, were fed either a standard diet, low Ca diet, low Ca-Mg diet, or low Ca-Mg diet with high Al for 90 days. Ca, Mg and Al content was determined in central nervous system (CNS) tissues and bone using inductively coupled plasma emission spectrometry (ICP). In separate studies, five male Japanese macaques (Macaca fuscata), weighing 3.5 to 5 kg, were fed alternately with diets, normal in Ca, low in Ca, low in Mg, low in Ca-Mg, or low in Ca-Mg with added Al for four-week periods. Serum Ca, Mg, Al, parathyroid hormone (PTH), bone Gla-protein (BGP) and alkaline phosphatase (ALP) were measured after feeding each dietary regimen. Ca and Mg levels in lumbar vertebrae and femur were significantly reduced and bone Al levels were significantly increased in rats fed diets deficient in Ca alone or diets low in Ca-Mg with or without added Al. Al content in bones was also higher in rats fed the Ca deficient diets. In monkeys fed the low Ca-Mg diet with added Al, reduced levels of serum Ca and Mg, serum PTH, BGP, and ALP were apparent. Our data support the conjecture that deranged bone mineralization induced by chronic dietary deficiency of Ca accelerates mobilization of Ca and Mg from bone and deposition in brain.

High aluminum deposition in the central nervous system of patients with amyotrophic lateral sclerosis from the Kii Peninsula, Japan: two case reports.

We report two cases of amyotrophic lateral sclerosis (ALS), in which metal analysis revealed markedly higher aluminum concentration in the central nervous system (CNS) as well as higher calcium and lower magnesium concentration and higher Ca/Mg ratios compared with controls. Case 1 was a 55-year-old housewife and total duration of illness was 2 years and 2 mon from onset of clinical symptom. Case 2 was a 80-year-old woman and total duration of illness was 10 mon. Results showed that neither were exposed to toxic environments nor any neurologic disease in the past history. Postmortem examination disclosed motor neuron death and degeneration of pyramidal tracts. The significance of metal metabolism in pathogenesis of amyotrophic lateral sclerosis is discussed.

Distribution of magnesium in central nervous system tissue, trabecular and cortical bone in rats fed with unbalanced diets of minerals.

Recent epidemiological changes in patterns of foci of amyotrophic lateral sclerosis (ALS) in the Western Pacific suggest that environmental factors play a contributory role in the pathogenic process of this disorder. In this experimental study on rats, a similar situation of dietary mineral imbalance was created as is found in the soil and drinking water of these ALS foci with a low content of calcium (Ca) and magnesium (Mg) and a high content of aluminum (Al). In groups of rats fed a low Ca diet, low Ca-Mg diet, and low Ca-Mg plus high Al diet, serum Ca levels were found to be lower than those in a group fed a standard diet. Also, serum Mg levels were lower in the groups fed a low Ca-Mg diet and a low Ca-Mg plus high Al diet than in the groups fed a standard diet and only a low Ca diet. There was no significant difference in Mg content of central nervous system (CNS) tissues of groups fed unbalanced and standard diets, except for a significant decrease in Mg content of the spinal cord of rats fed a low Ca-Mg plus high Al diet. Mg content of the lumbar spine and cortical bone decreased in the unbalanced diet groups compared with that of a group fed a standard diet. These findings suggest that under the disturbed bone mineralization induced by unbalanced mineral diets, Mg may be mobilized from bone to maintain the level necessary for vital activity in soft tissues including CNS tissue.