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Biomedical subjects

K Yamane

Publications and source records attributed to K Yamane.

At least 55 records · Page 3Linked to original sources

Clinical significance of surfactant protein D as a serum marker for evaluating pulmonary fibrosis in patients with systemic sclerosis.

OBJECTIVE: To determine the clinical significance of surfactant protein D (SP-D), a useful marker for evaluating various lung diseases, in patients with systemic sclerosis (SSc) and to clarify any clinical significance between SP-D and KL-6, which is known to be correlated with pulmonary fibrosis (PF) in SSc patients. METHODS: We used a specific enzyme-linked immunosorbent assay to measure serum SP-D levels in 83 patients with SSc and 31 healthy control subjects. RESULTS: The serum levels of SP-D were significantly higher in patients with SSc than in healthy controls (mean +/- SD 81.9+/-59.2 versus 34.8+/-13.7 ng/ml). Serum SP-D levels in patients with diffuse cutaneous SSc were significantly higher than those in patients with limited cutaneous SSc (98.8+/-72.1 versus 66.8+/-40.0 ng/ml). Serum SP-D levels in patients with PF were significantly elevated compared with those in patients without PF (99.7+/-64.1 versus 65.3+/-49.4 ng/ml). Moreover, the incidences of decreased percentage diffusing capacity for carbon monoxide and decreased percentage vital capacity were also significantly greater in patients with elevated SP-D levels than in those with normal levels (67% versus 43% and 36% versus 17%, respectively). There was a significant positive correlation between serum levels of SP-D and KL-6. Serum SP-D and KL-6 levels showed almost the same sensitivities and specificities in the diagnosis of PF (68% versus 73% and 70% versus 74%, respectively). These two markers also predicted PF to almost the same degree (31% versus 33%, respectively). CONCLUSION: These results suggest that SP-D, as well as KL-6, may be a useful serum marker for evaluating PF in patients with SSc.

Antibodies, Antinuclear↗

Immunohistochemical and ultrastructural study of basophilic inclusions in adult-onset motor neuron disease.

We immunohistochemically and ultrastructurally studied basophilic inclusions (BI) in a patient with adult-onset sporadic motor neuron disease (MND). BI were frequently observed not only in degenerated anterior horn cells, such as central chromatolytic neurons, but also in normal-appearing large anterior horn neurons. They had various shapes, round, elliptical or irregular, and occasionally they had distinct basophilic rims. They also varied in size. There were no halos around them nor core in their centers. Immunohistochemically, some BI were immunostained for ubiquitin or SOD1, but BI were not immunoreactive with anti-phosphorylated neurofilament (SMI 31), phosphorylated tau, cystatin C or Golgi (MG-160) antibodies. Ubiquitin-positive skein-like inclusions (SI) were occasionally observed in the somata of anterior horn neurons. Ultrastructurally, BI consisted of filamentous structures associated with granules, which were attached to thick filaments. The thick filaments were straight without constriction or side arms and their diameter was twice that of the neurofilaments. BI occasionally contained tubular structures among the granule-associated filaments. The granulo-filamentous profiles varied from being compactly arranged to being more loosely packed. The structure of BI resembles that of the Lewy body-like hyaline inclusions (LBHI) observed in sporadic MND patients. Bundles of filaments resembling SI, which were composed of compactly packed filaments without fine granules running parallel to the longitudinal axis, were frequently observed inside or at the periphery of BI, and occasionally clustered in the perikarya. Each filament measured approximately 15-25 nm in diameter, and a bundle of these grouped filaments was sometimes surrounded by a unit membrane. We also occasionally observed in-between structures of BI and bundles of filaments resembling SI. These findings suggest a certain relationship between BI, SI and LBHI in the pathomechanism of BI development. Further studies are needed to elucidate whether sporadic adult-onset MND characterized by BI forms a different subtype of MND.

Anterior Horn Cells↗

Anti-U1RNP antibodies in patients with localized scieroderma.

Antibodies to U1 ribonucleoproteins (RNP) have been detected in serum from patients with various autoimmune diseases. However, the presence of anti-U1RNP antibodies in patients with localized scleroderma has not been reported. In this study, we examined the frequency of anti-U1RNP antibodies using immunoprecipitation of U small nuclear RNAs and determined the antigen specificity by immunoblotting. Of 70 serum samples from patients with localized scleroderma, 2 (3%) immunoprecipitated U1 small nuclear RNA. Indirect immunofluorescence using HEp-2 cells as substrate showed coarse speckled nuclear fluorescence without nucleolar staining in both of the samples positive for anti-U1RNP antibodies. In addition, the presence of anti-U1RNP antibodies in each serum sample was confirmed by immunodiffusion against HeLa cell extracts. Immunoblotting analysis showed anti-70 kDa antibodies in each serum sample. This reaction against 70 kDa protein in the patients with localized scleroderma was analogous to that in patients with systemic sclerosis or mixed connective tissue disease. Both patients with positive serum were diagnosed as having linear scleroderma, but neither had evidence of Raynaud's phenomenon or sclerodactyly. These results indicate that the presence of anti-U1RNP antibodies is one of the serological abnormalities in localized scleroderma, and that the mechanism of induction of anti-U1RNP antibodies in patients with localized scleroderma might be similar to that in patients with systemic sclerosis and mixed connective tissue disease.

Autoantibodies↗

B-mode ultrasonographic investigations of morphological changes in endarterectomized carotid artery.

BACKGROUND: The aim of this study was to assess morphological changes in the endarterectomized carotid region using B-mode ultrasonography (USG). METHODS: USG examinations were performed on 54 Japanese patients who underwent carotid endarterectomy (CEA). The endarterectomized regions were periodically investigated and the intima-media thickness (IMT) was measured. RESULTS: Postoperative USGs revealed an evident step formation (type I: 20.4%), a gentle slope (type II: 42.6%), or complete smoothness (type III: 37.0%) at the junction of the endarterectomized carotid region and the common carotid artery (CCA). The IMT of the CCA progressively increased from type I to type III. Patients were followed up for an average of 2.7 years. The new intima-media complex (IMC) was confirmed in all cases after 9 months; it was visualized as an isoechoic layer (7%) or a mixture of iso- and hypoechoic layers (93%). Changes in the IMT during the follow-up period were classified into three groups: no change (group I: 23.1%), decrease (group II: 15.4%), and increase (group III: 61.5%). CONCLUSIONS: USG is useful to clarify the complicated healing processes of the endarterectomized carotid artery.

Adult↗

A simple titration method for determining Chlamydia infectivity using an image analyzing system.

In the biological study of Chlamydia, it is very important to determine the infectivity titer of the organism. For many years researchers used the serial dilution method to determine this titer. This method consists of diluting the material to be examined, inoculating suitable dilutions into susceptible cell cultures and cultivating them. The number of inclusions formed in host cells can be calculated with the naked eye under a microscope. The precision and accuracy of this method, however, depend on the number of inclusions per field and the number of fields counted. In this report, we present a simple and rapid method for counting a large number of inclusions using an image analysis system and an appropriate number of samples, and propose a sampling method based on a statistical analysis of the data obtained with 84 microscopic fields.

Animals↗

Prevalence in myositis of antibodies recognizing anti-U3 RNA probably in a novel complex with 22/25 kD protein and not fibrillarin.

New antibodies against a U3 snRNP, which were named anti-Myo 22/25 antibodies, were detected in four (8%) of 53 serum samples from patients with polymyositis/dermatomyositis (PM/DM) by RNA immunoprecipitation. In the protein immunoprecipitation analysis, all four serum samples precipitated 22 kDa and 25 kDa proteins, which were not precipitated by normal serum or serum positive for antifibrillarin antibodies. Three of the four PM/DM patients had other identified autoantibodies including anti-PL-12 antibodies, antihistone antibodies (AHA), anti-SS-A antibodies and anti-SS-B antibodies defined by double immunodiffusion, ELISA or RNA immunoprecipitation, although there were no significant correlations between anti-Myo 22/25 antibodies and clinical or laboratory findings. There may be a subgroup of PM/DM patients whose sera are positive for anti-Myo 22/25 antibodies.

Adolescent↗

Crystal structure of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. 1011 complexed with 1-deoxynojirimycin at 2.0 A resolution.

1-Deoxynojirimycin, a pseudo-monosaccharide, is a strong inhibitor of glucoamylase but a relatively weak inhibitor of cyclodextrin glucanotransferase (CGTase). To elucidate this difference, the crystal structure of the CGTase from alkalophilic Bacillus sp. 1011 complexed with 1-deoxynojirimycin was determined at 2.0 A resolution with the crystallographic R value of 0.154 (R(free) = 0.214). The asymmetric unit of the crystal contains two CGTase molecules and each molecule binds two 1-deoxynojirimycins. One 1-deoxynojirimycin molecule is bound to the active center by hydrogen bonds with catalytic residues and water molecules, but its binding mode differs from that expected in the substrate binding. Another 1-deoxynojirimycin found at the maltose-binding site 1 is bound to Asn-667 with a hydrogen bond and by stacking interaction with the indole moiety of Trp-662 of molecule 1 or Trp-616 of molecule 2. Comparison of this structure with that of the acarbose-CGTase complex suggested that the lack of stacking interaction with the aromatic side chain of Tyr-100 is responsible for the weak inhibition by 1-deoxynojirimycin of the enzymatic action of CGTase.

1-Deoxynojirimycin↗

Antibodies to Th/To ribonucleoprotein in patients with localized scleroderma.

OBJECTIVE: Antibodies to Th/To ribonucleoprotein (anti-Th/To) have been detected almost exclusively in patients with systemic sclerosis. Therefore, we aimed to determine the prevalence of anti-Th/To in patients with localized scleroderma. METHODS: Seventy serum samples from patients with localized scleroderma were examined by RNA immunoprecipitation and indirect immunofluorescence analysis using HEp-2 cells as substrate. RESULTS: Three (4%) of 70 sera from patients with localized scleroderma immunoprecipitated 7-2 (Th) RNA and 8-2 (To) RNA. Indirect immunofluorescence analysis demonstrated that all the sera positive for anti-Th/To showed mainly nucleolar staining. In one patient, the coexistence of anti-histone antibody with anti-Th/To was detected by enzyme-linked immunosorbent assay for anti-histone antibody and confirmed using an absorption test with histones followed by indirect immunofluorescence analysis. Moreover, the localized scleroderma patients with anti-Th/To tended to have significantly fewer sclerotic lesions than those without. CONCLUSION: Anti-Th/To is one of the serological abnormalities in localized scleroderma, and the presence of anti-Th/To may be a serological indicator of a mild form of cutaneous involvement.

Antibodies↗

The successful treatment of prurigo pigmentosa with macrolide antibiotics.

Recent studies have demonstrated that macrolide antibiotics have anti-inflammatory as well as antibacterial effects. Therefore, macrolide antibiotics have been successfully used to treat patients with various inflammatory diseases. We evaluated the effect of macrolide antibiotics in 4 patients with prurigo pigmentosa who were treated with either 400 mg of clarithromycin or 300 mg of roxythromycin daily. Eruption and pruritus disappeared within a week in all the patients while those symptoms were unresponsive to other drugs. Although the mechanism of this effect remains unclear in patients with prurigo pigmentosa, macrolide antibiotics can be considered as an alternative treatment for prurigo pigmentosa.

Adult↗

Involvement of fluctuating high blood pressure in the enlargement of spontaneous intracerebral hematoma.

The correlations between changes in blood pressure after admission and hematoma expansion were investigated in 118 patients with spontaneous intracerebral hematoma admitted within 24 hours of onset who underwent serial computed tomography. Multiple logistic regression was performed to assess correlations between hematoma enlargement and clinical characteristics on admission. Hematoma enlargement was predominantly correlated with time of onset (p = 0.01567), and not well correlated with blood pressure at admission (p = 0.07908). Serial changes in blood pressure were investigated in 57 patients admitted within 6 hours of ictus whose blood pressures were monitored every hour from admission. Wilcoxon signed-rank analysis was used to determine the relationships between hematoma enlargement and blood pressure. Patients with hematoma enlargement was significantly correlated with increased blood pressure (p = 0.0004). Increases in blood pressure after admission may be a factor in hematoma enlargement.

Aged↗

[Hemodynamic effects of STA-MCA anastomosis on patients with occlusion of the main cerebral artery].

PURPOSE: We studied cerebral circulation in patients with occlusion of the main cerebral artery and investigated the efficacy of STA-MCA anastomosis. PATIENTS AND METHODS: Thirty-six patients with occlusion of the main cerebral artery were studied. Twenty-three patients had occlusion of the internal carotid artery and 13 had occlusion of the middle cerebral artery. The mean age was 62 years. Cerebral blood flow (CBF) was measured in all patients and cerebrovascular reactivity (CVR) was examined in 11 patients by xenon enhanced CT. Intraoperatively, cortical arterial pressure and anastomotic flow were measured. RESULTS: There was no perioperative mortality or morbidity. There was no ipsilateral stroke recurrence during the follow-up period averaging 35.1 months. Patency of the anastomosis was verified in 91% of the patients by magnetic resonance angiography. Twenty-three (64%) patients showed decreased CBF before the operation and 57% of these patients showed improvement to the normal range after STA-MCA anastomosis. All of the eight patients with decreased CVR showed improvement after the operation. Anastomotic flow correlated significantly with the cortical arterial pressure. CONCLUSION: STA-MCA anastomosis could improve cerebral circulation of patients with low CBF or low CVR due to occlusion of the main cerebral arterial. It was concluded that STA-MCA anastomosis may contribute to the reduction of stroke recurrence, if perioperative complications are reduced.

Aged↗

[Relationship between flow volume curve and CT findings in non-smoking patients with long histories of bronchial asthma].

AIM: This study was conducted to verify whether bronchial asthma (BA) alone causes pulmonary emphysema (PE), and to examine the computed tomography (CT) findings in non-smokers with BA demonstrating the flow volume curve (FV curve) characteristic of PE. SUBJECTS AND METHODS: Non-smoking patients with a history of BA for more than 20 years were divided into 2 groups: the dogleg pattern group (n = 5), with an FV curve characteristic of PE, and the concave pattern group (n = 16) with an FV curve characteristic of BA. CT scans was performed using CT values (level, 900 H.U.; width, 400 H.U.) that facilitate detection of a low attenuation area (LAA), and using conventional CT values (level, 700 H.U.; width, 1,300 H.U.). LAA (including air trapping), thickness of the bronchial wall, and partial atelectasis were compared between the 2 groups. RESULTS: PE was not detected, although air trapping was found in all subjects. The thickness of the airway was greater in the dogleg pattern than in the concave pattern. The incidences of air trapping and partial atelectasis were higher in the former than in the latter. DISCUSSION: BA alone may not cause PE. Some BA patients without PE show the FV curve characteristic of PE, reflecting an increase in the thickness of the airway wall and a decrease in the pulmonary ventilation probably due to the air trapping and the partial atelectasis.

Aged↗

[Diffusion-weighted imaging of brain death: study of apparent diffusion coefficient].

DWI(Diffusion-weighted images) of the brain has been revealed to be useful in diagnosis of several clinical conditions. However, little is known about DWI with regard to brain death. We had opportunities to study patients with brain death. Case 1. A 34-year-old woman experienced cardiopulmonary arrest due to severe ventricular fibrillation, and resuscitated after about 120 minutes. After brain death, DWI showed high signals in the cerebral cortex, putamen, thalamus, brain stem and cerebellum, and ADC(apparent diffusion coefficient) values were 30-40% lower than those of normal volunteers. Case 2. A 45-year-old woman experienced cardiopulmonary arrest due to pontine hemorrhage, and was resuscitated after about 20 minutes. Before brain death, DWI showed high signals in the cerebral cortex, putamen, thalamus, brain stem and cerebellum, and ADC values were the same as those of normal volunteers. After brain death, DWI showed more clearly defined in these areas, but ADC values were 30-40% lower than the first values. DWI and ADC mapping shows areas corresponding to edema of a cytotoxic nature and to ischemic tissue. The characteristic views of high signals in the whole area of the brain were present, and an objective evaluation was possible with DWI after the brain death diagnosis by measuring ADC values.

Adult↗

The BRCT regions of tumor suppressor BRCA1 and of XRCC1 show DNA end binding activity with a multimerizing feature.

The BRCT regions are two repeating structures in BRCA1 at the carboxyl-terminus and are ubiquitous in some proteins involved in cell cycle checkpoint and in DNA repair. Here, using electron microscopy, we show direct evidence that the BRCT regions of BRCA1 bound double-strand breaks of DNA. The BRCT regions could multimerize thus forming large protein particles. Smeared patterns of DNA fragments were consistently shown in the gel retardation assay. A single BRCT was sufficient for DNA binding. The smeared patterns were also observed in BRCTs of TopBP1, suggesting that multimerization may be an important feature of BRCTs. The recombinant second BRCT of XRCC1 (X-ray repair cross-complementing group 1), whose folding was determined by X-ray crystallography, also showed similar DNA end binding images. It is possible that some BRCTs are fundamental structures that detect DNA damages.

Amino Acid Sequence↗

Anaerobic glycolysis is crucial for the maintenance of neural activity in guinea pig hippocampal slices.

To investigate the functional significance of anaerobic and aerobic glycolysis on neural activity and levels of high energy phosphates, we tested the effects of glucose, mannose, fructose lactate, and pyruvate on the maintenance of neural activity and on the levels of ATP and creatine-P (CrP) in hippocampal slices. For an index of neural activity, population spikes (PS) evoked in the granule cell layer were monitored. Immediately after deprivation of glucose, the PS amplitude was gradually reduced and extinguished within 30 mm. Replacement of glucose with either lactate or pyruvate resulted in a decay and loss of PS with a similar time-course as observed during glucose deprivation. However, after the complete loss of neural activity for 10-20 min the PS reappeared and recovered to normal levels. The replacement of glucose with either mannose or fructose resulted in a transient decrease of the PS to 80-70% of the original amplitude in 20 mm, followed by recovery. The time-course of the decrease of PS in the mannose-containing medium was slower than that in the medium containing fructose and the time-course of recovery was faster. ATP and CrP were reduced to 90 and 70% of original level in each slice after glucose deprivation for 30 and 100 mm, respectively. In media containing either lactate, pyruvate, mannose, or fructose, the level of ATP and CrP was maintained at the original level. The anaerobic metabolic rate of glucose, mannose and fructose, determined by the rate of lactate production during complete anoxia, was consistent with the order of the decay and recovery of the PS in mannose and fructose-containing medium. The mode of the transient decay or loss of PS with no apparent reduction in the levels of ATP and CrP in each slice, in the presence of either mannose, fructose or lactate, indicates that anaerobic glycolysis is crucial for the maintenance of PS. The results obtained in this experiment are not in accordance with the reports by Schurr and Fowler in which they showed that lactate can support neural activity, although they did not measure the levels of ATP and CrP in slices. The present experiment disclosed that this discrepancy was due to the difference of slice preparation; namely rapidly prepared slice with shorter period anoxia (1 mm after removal of hippocampal tissue block) gives the results mentioned in the present study whereas slowly prepared slices by vibratome or chopper method with longer period of ischemia (5-20 mm), did not show transient loss of PS after application of lactate. Thus present experiment indicates that glycolytic process is essential for maintaining neural activity for physiological state of slices, if it is admitted to say that rapidly prepared slices is more physiological because they are exposed on shorter period of ischemia, and that more careful attention should be paid for the interpretation of the results of slice experiment according to the method of slice preparation.

Action Potentials↗

Involvement of 14-3-3 proteins in nuclear localization of telomerase.

Maintenance of telomeres is implicated in chromosome stabilization and cell immortalization. Telomerase, which catalyzes de novo synthesis of telomeres, is activated in germ cells and most cancers. Telomerase activity is regulated by gene expression for its catalytic subunit, TERT, whereas several lines of evidence have suggested a post-translational regulation of telomerase activity. Here we identify the 14-3-3 signaling proteins as human TERT (hTERT)-binding partners. A dominant-negative 14-3-3 redistributed hTERT, which was normally predominant in the nucleus, into the cytoplasm. Consistent with this observation, hTERT-3A, a mutant that could not bind 14-3-3, was localized into the cytoplasm. Leptomycin B, an inhibitor of CRM1/exportin 1-mediated nuclear export, or disruption of a nuclear export signal (NES)-like motif located just upstream of the 14-3-3 binding site in hTERT impaired the cytoplasmic localization of hTERT. Compared with wild-type hTERT, hTERT-3A increased its association with CRM1. 14-3-3 binding was not required for telomerase activity either in vitro or in cell extracts. These observations suggest that 14-3-3 enhances nuclear localization of TERT by inhibiting the CRM1 binding to the TERT NES-like motif.

14-3-3 Proteins↗