Toshiaki Inagaki

αB crystallin and HSP28 are enhanced in the cerebral cortex of patients with Alzheimer's disease

Abstract The localization of two small heat shock proteins, αB crystallin and 28-kDa heat shock protein (HSP28), was studied immunochemically and immunohistochemically in cerebral cortex of patients with Alzheimers disease (AD) and control groups. By specific immunoassays for αB crystallin and HSP28, we found that the concentrations of αB crystallin are elevated in the temporal and frontal lobes, while those of HSP28 are elevated in the temporal, frontal, and parietal lobes in the AD brains. Immunohistochemically, αB crystallin was localized in astrocytes and oligodendrocytes, while HSP28 was present in degenerating neurons in the cerebral cortex of both AD and control brains. However, αB crystallin-immunoreactive astrocytes and HSP28-immunoreactive degenerating neurons were more frequently observed in AD brains. The immunoreactivity for HSP28 was also found in the senile plaques in the AD brains. These findings suggest that the increased accumulations of these small heat shock proteins appear to be part of reactive processes of glial cells and neurons under pathologic conditions.

Lipid peroxidation and advanced glycation end products in the brain in normal aging and in Alzheimer's disease

Abstract. The cellular distribution of malondialdehyde (MDA) was assessed immunohistochemically in brain specimens from young and normal elderly subjects as well as patients with Alzheimers disease (AD). MDA was increased in the cytoplasm of neurons and astrocytes in both normal aging and AD, but was rarely detected in normal young subjects. By electron microscopic immunohistochemistry, neuronal MDA formed cap-like linear deposits associated with lipofuscin, while glial MDA deposits surrounded the vacuoles in a linear distribution. In the hippocampus, neuronal and glial MDA deposition was marked in the CA4 region but mild in CA1. By examination of serial sections stained with anti-MDA and antibodies against an advanced glycation end product, Nε-(carboxymethyl)lysine (CML), neuronal and glial MDA deposition was colocalized with CML in AD, but only neuronal MDA was colocalized with CML in normal aged brains. Glial MDA, although abundant in the aged brain, typically was not colocalized with CML. In AD cases, MDA was colocalized with tau protein in CA2 hippocampal neurons; such colocalization was rare in CA1. MDA also was stained in cores of senile plaques. Thus, while both MDA and CML accumulate under oxidative stress, CML accumulation is largely limited to neurons, in normal aging, while MDA also accumulates in glia. In AD, both MDA and CML are deposited in both astrocytes and neurons.

Five-year survival of older people with anemia: variation with hemoglobin concentration.

OBJECTIVES: To investigate the significance of low hemoglobin concentration and longevity in older people.

Age-related changes in human spinal ventral horn cells with special reference to the loss of small neurons in the intermediate zone: a quantitative analysis.

Abstract A cytoarchitectonic study of spinal ventral horn cells was performed to identify age-related changes. The diameter distribution of ventral horn neurons of the fourth lumbar segment of the spinal cord and their size and topographical distributions were investigated in 14 autopsy cases. These cases represented patients of 18–100 years of age who had died of non-neurological diseases. The results indicate that small neurons widely distributed in the intermediate zone of the ventral horn significantly diminished with aging (P < 0.0005, r = –0.898), whereas medium-sized and large neurons located in the medial and lateral nuclei showed only a slight decrease with advancing age. The total number of neurons in the whole ventral horn was also noted to decrease significantly with aging (P < 0.0005, r = –0.899). While small neurons in the intermediate zone of the ventral horn are thought to be mostly interneurons, their physiological function still remains obscure in many respects. The findings of this study provide insight into age-related cell loss in terms of size and location.

Advanced glycation end products co-localized with astrocytes and microglial cells in Alzheimer's disease brain

Abstract   In the previous study [Takeda et al. (1996) Neurosci Lett 221: 17–21], we reported that the advanced glycation end products (AGEs) in the external space of neuronal perikarya (extraneuroperikaryal AGE deposits) were significantly abundant in the Alzheimer’s brain. In this study, we investigated the spatial relationship of the extraneuroperikaryal AGE (carbocymethyllysine and pentosidine) deposits in astrocytes and microglial cells in the Alzheimer’s disease brain using double immunolabelling for AGEs and astrocyte or microglial cell markers. Most of the extraneuroperikaryal AGE deposits were co-localized with glial fibrillary acidic protein-positive astrocytes. AGE deposit-bearing astrocytes also contained Gomori-positive granules. Furthermore, some of the extraneuroperikaryal AGE deposits were co-localized with microglial cells. These extraneuroperikaryal AGEs may activate astrocyte and microglia, and play a role in pathogenesis of Alzheimer’s disease.

Developmental and Age-Dependent Changes of 28-kDa Calbindin-D in the Central Nervous Tissue Determined with a Sensitive Immunoassay Method

Abstract: For the quantitative analysis of vitamin D‐dependent 28‐kDa calcium‐binding protein (calbindin‐D) in the CNS, we have established a highly sensitive immunoassay method. The antisera were raised in rabbits with purified calbindin‐D from rat kidneys, and the antibodies were purified with a calbindin‐D‐coupled Sepharose column. The purified antibodies were specific for calbindin‐D, showing a single band on the immunoblot with the extract of rat kidney or cerebellum. The sandwich‐type immunoassay system was prepared by the use of purified monospecific antibodies, and the minimum detection limit of the assay was 0.1 pg or 3.6 amol of calbindin‐D, which was sufficiently sensitive for the measurement of calbindin‐D content in isolated Purkinje cell bodies at the level of single cells. The average content of calbindin‐D in a single Purkinje cell was 0.05 pg. Calbindin‐D was detected in most of the rat tissues examined, but it was present predominantly in the kidney and CNS, especially in the cerebellum. Calbindin‐D was detected at a similarly low level in the cerebral cortex, cerebellum, and brainstem of rat embryos of 15 gestational days, and it increased gradually but differently in these regions, reaching the respective adult levels by 4–5 weeks of postnatal age. In contrast, kidney calbindin‐D increased sharply between 15 gestational days and 3 postnatal days, reaching the adult level by 6 days of age. Calbindin‐D levels in the adult rat CNS were affected little by age, whereas the concentrations in human cerebral cortices were significantly low in the aged brain as compared with those in the young brain. However, the concentrations in various regions of cerebral cortex from patients with Alzheimers disease showed values similar to those in the relevant regions of the age‐matched control patients.

Concentrations of several proteins characteristic of nervous tissue in cerebral cortex of patients with alzheimer’s disease

Concentrations of nervous tissue-related proteins, including S-100 proteins (α and β), enolase isozymes (α and γ), superoxide dismutase (SOD) isozymes (Cu/Zn SOD and Mn SOD), and GTP-binding proteins (α subunits of GO and Gi2) were determined in the four cerebrocortical regions (superior frontal gyrus of frontal lobe, parahippocampal gyrus of temporal lobe, superior parietal lobule of parietal lobe, and calcarine area of occipital lobe) of patients with Alzheimer’s disease, and age-matched control and young control patients by means of enzyme immunoassay methods. Although the temporal cortex of some patients with Alzheimer’s disease (4/7) showed apparently enhanced S-100β with decreased γ-enolase, concentrations of neuronal (neuronspecific γ-enolase and the α subunit of GO) and glial (S-100β, S-100α, and α-enolase) marker proteins, and both SODs in each region were not significantly different between patients with Alzheimer’s disease and the agematched controls. Concentrations of Gi2α also showed similar values in the cerebral cortices of young and aged controls and patients with Alzheimer’s disease. However, when compared with young controls, S-100β in the four regions of patients with Alzheimer’s disease and aged controls, and Cu/Zn SOD in frontal cortex of patients with Alzheimer’s disease were significantly enhanced (P<0.01).

Neuronal and glial advanced glycation end product [Nε-(carboxymethyl)lysine] in Alzheimer's disease brains

Abstract. The cellular distribution of an advanced glycation end product [Nε-(carboxymethyl)lysine (CML)] in aged and Alzheimers disease (AD) brains was assessed immunohistochemically. CML was localized in the cytoplasm of neurons, astrocytes, and microglia in both aged and AD brains. Glial deposition was far more marked in AD brains than in aged brains, and neuronal deposition was also increased. On electron microscopic immunohistochemistry, neuronal CML formed granular or linear deposits associated with lipofuscin, and glial deposits formed lines around the vacuoles. Neuronal and glial deposits were prominent throughout the cerebral cortex and hippocampus, but were sparse in the putamen, globus pallidus, substantia nigra, and cerebellum, with glial deposits being far more prominent in AD brains. The distribution of neuronal and glial deposits did not correspond with the distribution of AD pathology. The extent of CML deposits was inversely correlated with neurofibrillary tangle formation, particularly in the hippocampus. Most hippocampal pyramidal neurons with neurofibrillary tangles did not have CML, and most of the neurons with heavy CML deposits did not have neurofibrillary tangles. In the hippocampus, neuronal CML was prominent in the region where neuronal loss was mild. These observations suggest that CML deposition does not directly cause neurofibrillary tangle formation or neuronal loss in AD.

Bibliographical investigation of complementary alternative medicines for osteoarthritis and rheumatoid arthritis

Background:  A variety of complementary and alternative medicine (CAM) treatments are provided to the elderly. We investigated the efficacy and safety of CAM substances that are available to patients with osteoarthritis (OA) and rheumatoid arthritis (RA), both in Japan and overseas.

Immunoreactive parvalbumin concentrations in parahippocampal gyrus decrease in patients with alzheimer's disease

By using a sensitive enzyme immunoassay system for rat parvalbumin, we determined parvalbumin contents in the 4 cerebrocortical regions (superior frontal gyrus of frontal lobe, parahippocampal gyrus of temporal lobe, superior parietal lobule of parietal lobe, and calcarine area of occipital lobe) of patients with Alzheimers disease and age-matched controls. Among the 4 regions, concentrations of parvalbumin were the highest in calcarine area (68.6 +/- 6.7 ng/mg protein, rat parvalbumin equivalents, mean +/- SE) and the lowest in the parahippocampal gyrus (11.0 +/- 1.7 ng/mg protein) in the controls. A similar regional difference of the concentration was observed also in the patients with Alzheimers disease. When compared with the controls, however, concentrations of parvalbumin in parahippocampal gyrus of patients with Alzheimers disease (4.0 +/- 0.9 ng/mg protein) were significantly low (P less than 0.01), showing less than a half of the control values. In contrast, the concentrations in the 3 other regions showed little difference between Alzheimers disease and the controls.