In vivo airway eosinophil accumulation induced by polymyxin-B reduces bronchial responsiveness in guinea pigs.

BACKGROUND: Chronic desquamative eosinophilic bronchitis and bronchial hyperresponsiveness have been considered essential for bronchial asthma. However, it has not been studied whether airway eosinophils enhance or inhibit bronchial responsiveness in vivo. OBJECTIVE: This study was conducted to elucidate the influence of airway eosinophil accumulation on bronchial responsiveness in vivo. MATERIALS AND METHODS: Guinea pigs were transnasally treated with 75 microg/kg of polymyxin-B or vehicle twice a week for a total of 3 weeks. Guinea pigs were surgically cannulated and artificially ventilated 24 h after the last administration of polymyxin-B or vehicle. Ten minutes after the installation of artificial ventilation, ascending doses of methacholine, acetylcholine or histamine were inhaled for 20 s at intervals of 5 min. Subsequent study was conducted 20 min after treatment of 60 mg/kg of indomethacin in the same manner. Final study was conducted in naive guinea pigs after single inhalation of 75 microg/mL of polymyxin B. RESULTS: The proportion of eosinophils in bronchoalveolar lavage fluid significantly increased in guinea pigs treated with polymyxin-B compared with vehicle. Bronchial responsiveness to inhaled methacholine, acetylcholine and histamine was significantly decreased by the polymyxin-B treatment. This protective effect induced by polymyxin B was abolished by pretreatment of indomethacin. A significant increase in bronchial responsiveness was observed after a single inhalation of polymyxin B. CONCLUSION: These results suggest that in vivo airway eosinophils may reduce non-specific bronchial responsiveness through inhibitory or bronchoprotective prostanoids.

Parkinson's disease associated with argyrophilic grains clinically resembling progressive supranuclear palsy: an autopsy case.

A 70-year-old male began to show akinesia, rigidity of extremities, finger tremor, disturbed vertical external ocular movement, and nuchal dystonia, which progressed slowly. Brain CT scan and magnetic resonance images showed slight atrophy of the frontal lobe and slight enlargement of the lateral ventricles. Hasegawa's dementia rating scale-revised version gave a moderate score of 11/30 points. He died of pneumonia at the age of 76. The clinical diagnosis was progressive supranuclear palsy (PSP). However, there were no neuropathological characteristics of PSP. Neuropathologically, Parkinson's disease was diagnosed. In addition, many argyrophilic grains (ArGs) in the gray matter were stained, especially in the insula, amygdala, hippocampus, parahippocampal gyrus, lateral occipitotemporal gyrus, and substantia nigra, by the Gallyas-Braak method. We consider that ArGs could modify the symptoms of Parkinson's disease and that Parkinson's disease with ArGs may show a PSP-like clinical course.

Influence of alpha-adrenoceptor blockade on antigen- and propranolol-induced bronchoconstriction in guinea-pigs in vivo.

1. Beta-adrenoceptor antagonists, such as propranolol, can provoke severe bronchoconstriction only in asthmatic subjects. Recently, we developed a guinea-pig model of propranolol-induced bronchoconstriction (PIB) and the purpose of this study was to investigate the role of alpha-adrenergic nerve pathways in this reaction. 2. Phentolamine administered after an antigen challenge did not inhibit PIB; however, its administration before the antigen challenge significantly inhibited the antigen-induced bronchoconstriction and also bronchoconstriction induced by methacholine inhalation. 3. We conclude that the alpha-adrenergic nerve system is not involved in the development of PIB following allergic reaction in our guinea-pig model.

Effect of thromboxane synthase inhibitor, CS-518, on propranolol-induced bronchoconstriction in guinea pigs.

Beta-adrenoreceptor antagonists, such as propranolol, can provoke severe bronchoconstriction in asthmatic subjects. Recently we developed an animal model of propranolol-induced bronchoconstriction and investigated the involvement of chemical mediators in this reaction. The purpose of this study was to elucidate the role of thromboxane A2 in the development of propranolol-induced bronchoconstriction after allergic bronchoconstriction. Passively sensitized guinea pigs were anesthetized and treated with diphenhydramine hydrochloride and were then artificially ventilated. Propranolol at a concentration of 10 mg/ml was inhaled 20 min after an aerosolized antigen challenge. A potent and selective thromboxane A2 synthase inhibitor, CS-518, in doses of 0.01, 0.1 and 1 mg/kg and vehicle were administered intravenously 15 min after the antigen challenge. Another study was performed in naive guinea pigs; ascending doses of methacholine (12.5, 25, 50, 100 and 200 microg/ml) were inhaled for 20 sec at 5-min intervals, 10 min after intravenous administration of CS-518. Propranolol inhaled 20 min after the antigen challenge caused bronchoconstriction in sensitized guinea pigs. CS-518 administered 15 min after the antigen challenge significantly inhibited propranolol-induced bronchoconstriction in a dose-dependent manner, while CS-518 did not influence the dose-dependent response to inhaled methacholine in naive guinea pigs. We conclude that thromboxane A2 contributes to the development of propranolol-induced bronchoconstriction following allergic reaction in our guinea pig model.

Comparison between multiple lacunar infarcted patients with and without dementia in nursing homes in shimane prefecture, Japan.

We evaluated the activity of daily living (ADL) scores, the frequency of hypertension (HT) and diabetes mellitus (DM), and the frequency and distribution of white matter lesions in a vascular dementia group with multiple lacunar infarctions (VD group; 20 cases) and a nondemented group with multiple lacunar infarctions (non-D group; 32 cases), relative to a normal control group (29 cases). There were no significant differences in HT and DM among the three groups. ADL scores were significantly lower in the VD than in the non-D group, each of which was lower than in the control group. Frequency of white matter lesions (Binswanger's disease-like lesions in the frontal and parietal lobes) was significantly higher in the VD than in the non-D group, which was higher than in the control group. We believe that for dementia with multiple lacunar infarctions, diffusely affected cerebrum including white matter lesions, which results in low ADL scores for symptoms, may be a prerequisite.

Dementia of Alzheimer type with and without multiple lacunar infarctions: evaluation of white matter lesions.

The white matter lesions in dementia of Alzheimer type (DAT) with and without multiple lacunar infarctions were studied relative to a normal control group. The frequency and distribution of white matter (WM) lesions in DAT (22 cases; mean age +/- standard deviation (SD), 88.1 +/- 5.8), DAT with multiple lacunar infarctions (DAT + CVD, 18 cases; mean age +/- SD, 87.8 +/- 6.0), and in a normal control group (17 cases; mean age +/- SD, 85.2 +/- 4.8) were evaluated. The frequency of myelin pallor (frontal, parietal and occipital lobes) was significantly higher in the DAT + CVD group than in the other groups (DAT and controls). There was no significant difference in the frequency of myelin pallor between the DAT and control groups. Therefore, it was concluded that the WM lesions in DAT are the result of ischemia rather than wallerian degeneration.

ADP-ribosylation of tubulin by chicken NAD-arginine ADP-ribosyltransferase suppresses microtubule formation.

We obtained evidence that tubulin and actin, two major cytoskeletal proteins, are preferentially ADP-ribosylated in the bovine brain cytosol by NAD-arginine ADP-ribosyltransferase purified from chicken polymorphonuclear leukocytes. ADP-ribosylation of tubulin almost completely blocked self-assembly of the protein. The stoichiometry of ADP-ribose incorporation into unassembled and assembled tubulin was 6 and 2 mol/mol of tubulin, respectively. These findings suggest that sites of ADP-ribosylation in the unassembled tubulin molecule are crucial for tubulin assembly, and that covalently attached ADP-ribose moieties interfere with tubulin interaction by steric hindrance or conformational change. Thus, ADP-ribosylation may be involved in cytoskeletal organization in the brain via the modification of tubulin.

Suppression of cell adhesion and spreading activities of fibronectin by arginine-specific ADP-ribosyltransferase from chicken polymorphonuclear leukocytes.

Arginine-specific ADP-ribosyltransferase present in secretory granules of chicken polymorphonuclear leukocytes (so-called heterophils) was shown to be released into the extracellular space by secretagogues (Terashima et al., J. Biochem. 120 (1996) 1209-1215). In the present work, we examined fibronectin as an extracellular target protein of the released transferase. Fibronectin was ADP-ribosylated by purified transferase and stoichiometry of ADP-ribose incorporation into fibronectin was 1.0 mol/mol of fibronectin. Cell adhesion and spreading assays revealed that ADP-ribosylation of fibronectin markedly inhibited the adhesion activity of fibronectin. A proteolytic peptide map of ADP-ribosylated fibronectin demonstrated that the modification occurs in the cell binding domain of fibronectin. ADP-ribosylation of the RGD peptide suggests that the RGD sequence is the modification site in the domain. ADP-ribosylation of fibronectin in plasma means that fibronectin can probably serve as the substrate for extracellularly released ADP-ribosyltransferase in vivo. Thus, in the extracellular space, ADP-ribosyltransferase released from polymorphonuclear leukocytes may perhaps be involved in regulation of cell adhesion process by interfering with the activity of fibronectin.

GTP-dependent modification of a 21-kDa substrate with NAD+ in bovine brain soluble fraction is not ADP-ribosylation of small G-protein but tailing of tRNA.

Labeling of 21-kDa material was observed when bovine brain soluble fraction was incubated with [adenylate-32P]NAD+ in the presence of GTP. The 21-kDa substrate, slightly smaller than C3 substrate in size, was labeled even without C3 exoenzyme. GTP could be replaced by nucleoside triphosphates other than ATP while ATP inhibited the GTP-induced labeling of 21-kDa substrate. After incubation of the soluble fraction with [adenylate-32P]NAD+ in the presence of GTP, [32P]ADP and [32P]ATP were detected in addition to [32P]AMP and [32P]ADP-ribose while only the last two nucleotides were observed without GTP. The 21-kDa substrate was labeled with [alpha-32P]ATP even in the absence of GTP, suggesting adenylylation rather than ADP-ribosylation. The labeled 21-kDa substrate, was extractable by phenol, disappeared with RNase treatment but not with tryptic digestion. Alkaline treatment of the phenol extract yielded an equal mixture of 3'-[32P]CMP and 2'-[32P]CMP. From these results we concluded that the 21-kDa labeling is a result of tRNA tailing with [alpha-32P]ATP generated from the [32P]AMP moiety of [adenylate-32P]NAD+. Results from reconstitution experiments using enzymes and tRNA purified from bovine brain soluble fraction, which are involved in this pathway, confirmed our conclusion.

Immunohistochemical localization of ADP-ribosylarginine hydrolase in rodent CNS.

Polyclonal antibodies were generated against ADP-ribosylarginine hydrolase (AAH), using recombinant fusion protein of rat AAH and glutathione-S-transferase as a immunogen, and affinity-purified. Western blotting showed that the antibodies recognized in mouse brain homogenate a single protein with a molecular mass of 38 kDa, the expected size for mouse AAH. An analysis using the antibodies revealed that heavy labelings were apparent in various brain regions. In the cerebral cortex, pyramidal cells in layers III and V were the most heavily labeled. In the hippocampal formation, labeling was present on the pyramidal neurons and granule cells. The most heavily immunostained cell type was the pyramidal neuron of CA3. In the cerebellum, Purkinje cells were the most heavily labeled. Less intense staining was present over the granule cells. In the basal ganglia, neurons in the caudate nucleus and large multipolar cells in the amygdaloid complex were immunoreactive. Heavy labeling was seen in many midbrain and brainstem nuclei. Neurons in the habenula and ependymal cells were stained heavily. On Western blot analysis of rat cerebrospinal fluid (CSF), the anti-AAH antibodies recognized a protein with a molecular mass of 38 kDa. This is apparently the first evidence of a widespread but distinctive distribution of AAH in neurons of mouse brain and the presence of extracellular AAH in rat CSF.

ADP-ribosylation of Arg28 and Arg206 on the actin molecule by chicken arginine-specific ADP-ribosyltransferase.

We reported previously on ADP-ribosylation of actins by chicken arginine-specific ADP-ribosyltransferase in vitro and in situ and the inhibition of actin polymerization by this modification [Terashima, M., Mishima, K., Yamada, K., Tsuchiya, M., Wakutani, T. & Shimoyama, M. (1992) Eur. J. Biochem. 204, 305-311]. In the present study, we determined amino acid residues of ADP-ribosylation site(s) in globular (G-) and filamentous (F-) actins and examined the molecular basis of the modification of actin. Arginine-specific ADP-ribosylation occurred at Arg28 and Arg206 in G-actin, but only at Arg28 in F-actin. ADP-ribosylation of Arg206, located on the pointed end of the actin molecule, significantly blocked the interaction with deoxyribonuclease I. These results indicate that Arg206 in G-actin may be involved in actin polymerization. ADP-ribosylation of Arg28, located on the outer surface of actin molecule, did not affect the binding activity with myosin subfragment-1, that is thought to interact through the N-terminal amino acid residues of G-actin. ADP-ribosylation at both Arg28 and Arg206 of G-actin had no apparent effect on the intrinsic ATPase activity. We concluded from this study that ADP-ribosylation of Arg206 in G-actin causes the inhibition of actin polymerization, and that ADP-ribosylation of Arg28 occurs in F-actin.

Familial Creutzfeldt-Jakob disease: three autopsy cases of the panencephalopathic type.

Three autopsy cases of panencephalopathic type of familial Creutzfeldt-Jakob disease (CJD) were investigated. Cases 1 (51-year-old male) and 3 (54-year-old female) were siblings and Case 2 (68-year-old female) was their aunt. In cases 1 and 3, the age of onset (Case 1:51, Case 3:53), duration of illness (Case 1:9 months, Case 3:8 months) and neuropsychiatric symptoms (pyramidal and extrapyramidal tracts involvements, blindness and dementia in chronological order) were similar, but in Case 2, the onset was later (66 years old), duration was longer (32 months) and the initial symptom was dementia. Myoclonus and apallic state in the terminal stage were common to all 3 cases. Neuropathologically, all 3 cases had characteristics that indicated panencephalopathic type of CJD. Cases 1 and 3 had similar neuropathological findings with characteristic circumscribed necrotic foci in the subcortical white matter. In Case 2 in contrast, diffuse demyelination and fibrillary gliosis in the cerebral white matter were observed without circumscribed necrotic foci. In the cerebellum of Case 3, granular cell loss was very slight. The other lesions in the cerebral cortex and striatum of the 3 cases were common. In conclusion, the clinical symptoms and neuropathological findings of our familial CJD cases were different from one another.

Neuro-Behçet disease with demyelination and gliosis of the frontal white matter.

A rare case of neuro-Behçet disease with diffuse demyelination and gliosis of the frontal white matter is reported clinico-pathologically. The disease began with genital ulcer and recurrent oral aphthosis when the patient was 42 years of age. There was erythema, moderate fever, CSF (cerebrospinal fluid)-pleocytosis and elevated CSF-globulin. He was diagnosed as having neuro-Behçet disease and treated with prednisolone. He gradually became euphoric, disinhibited, indifferent and demented. His cranial CT showed diffuse low density areas in the bilateral frontal white matter. He became bedridden, akinetic mute and died from respiratory dysfunction 3 1/2 years after onset. The following neuropathological findings were observed: 1) Moderate demyelination and gliosis was present mainly in the frontal and parietal white matter. 2) There were many micro-spongious necrotic foci in the gray and white matters of the cerebrum, basal ganglia, thalamus, midbrain and pons, some of which were accompanied by gliosis. 3) From 1/2 to 1/3 of all micro-necrotic foci in the frontal white matter were old and accompanied by gliosis. The white matter containing numerous micro-necrotic foci had myelin pallor and gliosis. 4) There was neither micro-necrosis nor gliosis in the occipital lobe. The pathogenetic correlation of white matter lesions with primary and secondary circulatory disturbances is discussed.

An autopsy case of PSP with astrocytic inclusions.

Argyrophilic glial cytoplasmic inclusions were observed in astrocytes (Astrocytic Inclusions) in a case of progressive supranuclear palsy (PSP). The inclusions were predominantly distributed in the lenticular nucleus and midbrain, but not in the cerebral cortex, white matter nor in the thalamus. When examined by double staining with Gallyas silver and GFAP, they resembled the neurofibrillary tangles (NFT) and were tau positive. The distribution of these inclusions was not related to that of NFT. These inclusions indicate that astrocytes were involved in the same pathological changes as NFT. Argyrophilic inclusions were also observed in oligodendroglias on Gallyas silver staining. They appeared coiled and located mainly in the cerebral and cerebellar white matter.

ADP-ribosylation of myelin basic protein and inhibition of phospholipid vesicle aggregation.

Four isoforms of myelin basic protein (MBP) from chicken brain were ADP-ribosylated by chicken heterophil ADP-ribosyltransferase. The 21-kD isoform was the most preferential substrate of this transferase. With this isoform, the Km values were estimated to be 330 mumol/l for NAD and 30 mumol/l for MBP, and the optimal pH for ADP-ribosylation was 8.5. The stoichiometry of ADP-ribose incorporation into 21-kD MBP was 3.5 mol of ADP-ribose/mol MBP. We found the inhibition of ADP-ribosylation of MBP by hydroxylamine and L-arginine indicating that this modification was likely to be mediated by arginine residues. Proteolytic peptide maps of ADP-ribosylated MBP by chicken ADP-ribosyltransferase and cholera toxin showed partially different radio active bands. When 21-kD MBP was ADP-ribosylated by chicken transferase, the potential for phospholipid vesicle aggregation was reduced in proportion of the degree of ADP-ribosylation. The possibility that ADP-ribosylation of MBP may control stabilization of myelin through regulation of its affinity for phospholipid in vivo would need to be considered.

[An autopsy case of ataxic form of Creutzfeldt-Jakob disease].

An autopsy case of ataxic form of Creutzfeldt-Jakob disease (Brownell and Oppenheimer, 1965) was reported. The patient, a 71-year-old male, noticed ataxic gait at the beginning of June in 1988, and was admitted to the Hiroshima City Hospital for the neurological examination at the end of June. He showed ataxia of the left arm and legs and diplopia. Gradually he became delirious at night. On July 16, tremor-like involuntary movement of the left hand was noticed. On July 20, he became somnolent and doubly incontinent. Myoclonus and paratonic rigidity were also observed. The EEG showed periodic synchronous discharge on July 25. The brain CT and MRI were normal. He became apallic gradually and died on October 28. The duration of illness was 5 months. At autopsy, brain weighed 1000gr. Cerebral atrophy and slight enlargement of the ventricles were observed. The cerebellum was also slightly atrophic. Histologically, the destruction of the cerebral cortical layer, slight sieve-like spongy state of the neuropil, slight neuronal loss of the thalamus and sieve-like spongy state of the striatum were observed. The cerebellar lesion was the most severe, where granular cell loss and gliosis of the cortex were observed.

[Cases of numerous diffuse plaques in the neocortex but without severe senile changes in the hippocampus].

Using modified Bielschowsky method, we studied neuropathologically 159 aged subjects autopsied during the period from 1976 to 1988, of which we found 19 cases (average age at death: 82.6 ys) with numerous diffuse plaques in the frontal and temporal neocortex and no severe senile changes in H1-H3 of Ammon's horn (dp group). Amyloid angiopathy had been excluded and one case was excluded because of considerable cerebrovascular lesions. The dp group was divided into 8 demented (average age at death: 86.0 ys) and 10 nondemented patients (average age at death: 79.7 ys). We compared the number, type, and ratio of types of senile plaques in the frontal cortex, temporal cortex, and putamen of the demented and nondemented groups, and obtained the following results: (1) Eight (14%) of the 59 nondemented and 8 (40%) of the 20 demented cases in which no severe senile changes in the neocortex and hippocampus had been detected by Bodian stain showed numerous diffuse plaques in the neocortex when the modified Bielschowsky method was used. (2) The ratio of classic and primitive plaques to diffuse plaques in the frontal cortex was the same in both groups, but the nondemented group had exclusively diffuse plaques in the temporal cortex. (3) In the putamen 2 nondemented cases (20%) and 6 demented cases (75%) had exclusively diffuse plaques. We considered that classic and primitive plaques are more closely related to dementia than are diffuse plaques in the temporal lobe in cases without severe senile changes in the hippocampus.

[A case of Binswanger disease with numerous diffuse plaques in the neocortex].

A case of Binswanger disease with numerous diffuse plaques in the neocortex was reported. This male patient had a previous history of hypertension and myocardial infarction. From the age of 60, he developed dysarthria, bradykinesia, marche à petit pas and falling down. Neurological examination at his first admission disclosed muscular rigidity and increased jaw and deep tendon reflexes, but dementia was not found. Brain CT showed moderate brain atrophy and EEG consisted of slow wave dysrhythmia. He was diagnosed of Parkinsonism and treatment started without effects. During his second admission for the treatment of myocardial infarction, at the age of 64, delirium developed. Progressive dementia began and finally he was confined to bed. From the age of 69, spontaneous speech became almost lost. Contracture of the extremities, increased deep tendon reflexes and force grasping were noted. Brain CT showed symmetrical low attenuation in the frontal and parietal white matter with moderate dilatation of the lateral ventricles. At the age of 70, he died of general prostration about ten years after the initial symptoms. Neuropathological findings: Macroscopic findings: The brain weighed 1300 g. Atherosclerotic changes of the large arteries at the base of the brain were moderate. Coronal sections of the brain showed moderate enlargement of the lateral ventricles with multiple small lacunes in the basal ganglia. Microscopic findings: Bilateral diffuse demyelination of the white matter with sparing of the U-fibers was noted. Holzer stain revealed fibrillary gliosis in the left parietal and occipital white matter. Marked adventitial fibrosis of the deep white matter arteries and terminal stages of hyalinosis of the perforating arteries were found. Basal ganglia showed status lacunaris. Bilateral pyramidal tracts were atrophic secondly.(ABSTRACT TRUNCATED AT 250 WORDS)