Biomedical subjects

K Masuda

Publications and source records attributed to K Masuda.

At least 289 records · Page 16Linked to original sources

PubMed1996

FDG-PET in infectious lesions: The detection and assessment of lesion activity.

The usefulness of FDG-PET in the detection of infectious foci and the assessment of lesion activity was evaluated. The study covered 24 patients with 25 FDG-PET studies, including lesions of bacterial, tuberculous and fungal origins. The FDG uptake was determined by the lesion to muscle ratio (LMR) on the static images. The time activity curves (TACs) were classified into four patterns based on both the existence of an initial peak and a slope thereafter. A high FDG uptake was observed in 23 of 25 lesions (92%). Two lesions, in which no abnormal uptake was noted, included one in the healing stage and the other consisting of a cavity with a thin wall. The acute active lesions showed higher LMRs than the chronic active or healing lesions (mean +/- SD: 9.8 +/- 3.6, 3.6 +/- 1.8 and 4.3 +/- 1.7, respectively, p < 0.05), and they could be approximately distinguished by an LMR of 6. The patterns of the TACs in acute or chronic active lesions were either an increase without an initial peak or a plateau, while those in the healing lesions demonstrated predominantly an increase with an initial sharp peak. Our results indicated that FDG-PET is clinically useful in the detection of the infection of miscellaneous microorganisms as well as in the assessment of lesion activity.

Abscess↗

PubMed1996

A clinical evaluation of FDG-PET to assess the response in radiation therapy for bronchogenic carcinoma.

The clinical usefulness of FDG-PET in the prediction and assessment of response to radiation therapy in patients with bronchogenic carcinoma was evaluated. Thirty patients with untreated bronchogenic carcinoma were included in the study. All patients received FDG-PET before the initiation of radiation therapy, while 20 also received it after completing the therapy. The tumor to muscle ratio (TMR) was used as an index of the FDG uptake. The tumor response to therapy was classified as either a partial response (PR, n = 21) or no change (NC, n = 9) according to changes in the tumor size. Prognosis was made 6 months after the initiation of therapy, and was classified as either relapse (n = 19) or non-relapse (n = 9). The FDG uptakes both before and after therapy were compared with tumor response and prognosis. A high FDG uptake was noted in all 30 lesions before therapy. No significant differences in the uptake before therapy was observed according to the histological types nor T factors (UICC). The lesions with a higher uptake (TMR more than 7) responded better to therapy than those with a lower uptake (p < 0.05). The decrease in the uptake after therapy tended to be more prominent in the PR group than in the NC group. The rate of relapse was higher in lesions with a higher uptake before therapy (TMR more than 10) than in those with a lower uptake. The relapse group also showed a higher uptake after therapy than the non-relapse group. In addition, all 6 lesions showing a higher uptake (TMR more than 5) after therapy eventually relapsed (p < 0.05). Two lesions demonstrating a lower uptake both before and after therapy did not relapse, although no tumor regression due to the therapy was observed. These results indicate that FDG-PET plays a complementary role in both predicting and assessing the therapeutic response and prognosis in patients with bronchogenic carcinoma.

Adenocarcinoma↗

PubMed1996

Contrast-enhanced MRI in spinal arteriovenous malformations and fistulae before and after embolisation therapy.

MRI was performed in six cases of spinal arteriovenous malformation (AVM) and arteriovenous fistula (AVF) before and after embolisation. Intramedullary and perimedullary AVMs showed marked vascular enhancement after embolisation. This was thought to reflect feeding vessel occlusion and correlated well with a favourable clinical outcome. In dural AVFs, contrast-enhanced studies were essential for the diagnosis, unenhanced images being nonspecific. After embolisation, enhancement of the spinal cord was reduced, although one case with a poor outcome showed persistent enhancement.

Adult↗

PubMed1996

Differences in the reduced 18F-Dopa uptakes of the caudate and the putamen in Parkinson's disease: correlations with the three main symptoms.

It has been reported that the F-Dopa (FD) uptake in patients with idiopathic Parkinson's disease (PD) decreased significantly in the caudate and putamen when compared to controls. The FD uptake severely decreased in the putamen, while it was relatively spared in the caudate nucleus. We also previously reported that atypical parkinsonism with no or little tremor showed a homogeneously reduced FD uptake in both the caudate and the putamen. In this study we evaluated the caudate and the putaminal FD uptakes in relation to the three main symptoms in PD. The FD uptake was measured by PET with 6-L-[18F]fluorodopa in 17 patients with PD. The caudate and the putaminal FD uptake ratios to the cerebellum at 120 min were evaluated. The caudate and the putaminal FD uptake ratios in the patients with PD decreased as their clinical stages advanced. These decreases also correlated with the degree of rigidity and bradykinesia. However, such decreases did not correlate with the degree of tremor. The caudate-putamen index (CPI)(%), which was calculated by a formula based on the difference in the uptakes of the caudate and putamen divided by the caudate uptake, indicated 11.6 +/- 3.6, 16.5 +/- 5.5 and 18.3 +/- 4.1 in the group of no, mild and moderate tremor, respectively, and increased as the degree of tremor advanced. The CPI in the group of moderate tremor significantly increased from that in the group of no tremor (P < 0.04). However, the CPI did not correlate with the clinical stage, the degree of rigidity or the degree of bradykinesia. The FD/PET study therefore effectively demonstrated the severity of the clinical symptoms of rigidity and bradykinesia in patients with PD in correlation with a decrease in the FD uptakes in the caudate and the putamen, and it also demonstrated that the severity of tremor might have a different mechanism from that of such other symptoms as rigidity and bradykinesia.

Adult↗

PubMed1996

Positron-labeled antioxidant 6-deoxy-6-[18F]fluoro-L-ascorbic acid: increased uptake in transient global ischemic rat brain.

The in vivo uptake and distribution of 6-deoxy-6-[18F]fluoro-L-ascorbic acid (18F-DFA) were investigated in rat brains following postischemic reperfusion. Global cerebral ischemia was induced in male Wistar rats for 20 min by occlusion of four major arteries. Two time points were chosen for 18F-DFA injection to rats subjected to cerebral ischemia, at the start of recirculation and 5 days following recirculation. The rats were then killed at 2 h after tail-vein administration of 18F-DFA and tissue radioactivity concentration was determined. Increased uptake of radioactivity in particular brain regions, including the cerebral cortex, hypothalamus, and amygdala following injection of 18F-DFA, compared to the sham-operated control, was observed 5 days after reperfusion. Similar results were also obtained in in vitro experiments using brain slices. Abnormal in vivo accumulation of 45Ca, a marker of regional postischemic injury, was observed in these brain regions in tissue dissection experiments. Furthermore, metabolite analysis of nonradioactive DFA using 19F-NMR showed that DFA remained intact in the postischemic reperfusion brain. The present results indicate that 18F-DFA increasingly accumulates in damaged regions of postischemic reperfusion brain.

Animals↗

PubMed1996

Glucose metabolism in the cortical and subcortical brain structures in multiple system atrophy and Parkinson's disease: a positron emission tomographic study.

The brain glucose metabolism was studied by PET with 18F-FDG in 11 patients with multiple system atrophy (MSA) and 12 patients with idiopathic Parkinson's disease (PD). Seven of the 11 MSA patients were diagnosed as having olivopontocerebellar atrophy, two had striatonigral degeneration, while two demonstrated Shy-Drager syndrome. The glucose metabolic rates for each region in the PD patients showed no difference from the normal controls. The frontal, temporal and parietal cortical glucose metabolic rates and the caudate, the putaminal, the cerebellar and the brainstem glucose metabolic rates in the MSA patients decreased significantly from the controls. The atrophy of the cerebellum and the brainstem in the MSA patients were scored by MRI. The cerebellar and brainstem glucose metabolism in the MSA patients decreased as the atrophy score in such regions advanced in each group; however, some patients with no atrophy showed a decreased glucose metabolism. Although the cerebellar and the brainstem glucose metabolism decreased in all MSA patients, such a decrease was not observed in the SND patients. The decrease in the glucose metabolism for the non-cortical regions in the MSA patients seems to be due to a diffuse depletion of the neurons not restricted to the nigrostriatal neurons. Deafferentation to the cerebral cortices seems to result in a decreased cortical metabolism. The differences in the glucose metabolism between MSA and PD as assessed by PET may be caused by the pathophysiological differences between MSA and PD, and such differences therefore appear to be useful when making a differential diagnosis between MSA and PD. The relative sparing of the brainstem and cerebellar glucose metabolism is considered to be a feature of patients with SND.

Adult↗

PubMed1996

V alpha 14+ NK T cells: a novel lymphoid cell lineage with regulatory function.

A novel lymphoid lineage, NK T cells, was recently found. The NK T cells are the major population in the periphery comprising 5% of splenic T cells and 40% of bone marrow T cells. They express a unique TCR composed of invariant V alpha 14J alpha 281 and V beta 8.2 together with NK receptor (NKRPI). Surprisingly, the invariant V alpha 14+ TCR is exclusively expressed on NK T cells but not on conventional T cells. As the selective decrease in V alpha 14+ NK T cell population in the periphery is tightly correlated with autoimmune disease development, V alpha 14+ NK T cells control development of autoimmune diseases. We also found that V alpha 14 TCR gene rearrangement and transcripts were detected at an early embryogenesis (d9.5) before the thymus formation. Therefore NK T cells are in the distinct category from conventional T cells. The target of NK T cells is found to be CD1 (class 1b, monomorphic class I MHC-like molecule) present on bone marrow-derived cells and is killed by Fas-FasL interaction or perforin-mediated mechanisms. These results indicate that NK T cells consist of an immunoregulatory system different from defense system in terms of homogeneous repertoire, extrathymic development in early stage of gestation, and their regulatory functional role.

Animals↗

PubMed1996

Regulation of bcl-xL expression and Fas susceptibility in mouse B cells by CD40 ligation, surface IgM crosslinking and IL-4.

CD40 is one of the key molecules involved in the survival, growth and differentiation of B lymphocytes. In contrast, Fas (Apo-1, CD95) mediates apoptosis of a variety of cell types, including lymphocytes. Recent studies have found that Fas expression on mouse B cells could be strongly induced by CD40 ligation, a helper T cell-derived signal. Here, evidence is provided that CD40 ligation induced two distinct signals: one leading to the upregulation of Fas and the other leading to the enhanced Fas susceptibility. B lymphoma cell lines, CH31 and WEHI279, expressed Fas on cell surfaces, but were resistant to anti-Fas antibody (Ab) induced apoptosis. Treatment with CD40 ligand (CD40L), however, greatly enhanced Fas susceptibility of these cells. Similarly, normal splenic B cells became highly susceptible to Fas-mediated apoptosis following prolonged signaling through CD40. While CD40 ligation enhanced Fas-mediated apoptosis, stimulation with anti-IgM and IL-4 partially protected CD40L-activated B cells from Fas-mediated apoptosis. It was found that bcl-xL gene expression in normal splenic B cells was induced drastically by treatment with anti-IgM and IL-4, but not CD40L. By contrast, the expression of bcl-2 or bax was not significantly affected by these treatments. Moreover, in three of the four B lymphoma cell lines tested, Fas susceptibility correlated with the status of bcl-xL expression. The data suggest that an increase in bcl-xL expression may protect B cells from Fas-mediated apoptosis.

Animals↗

PubMed1996

Vimentin-positive adenocarcinomas of the stomach: co-expression of vimentin and cytokeratin.

Recently, the expression of vimentin has been reported in some carcinomas. This study was designed to clarify the significance of vimentin expression in solid type poorly differentiated adenocarcinomas of the stomach. Immunohistochemically, 239 poorly differentiated adenocarcinomas with solid components of the stomach were stained for vimentin. Vimentin-positive cases were also stained by CAM 5.2 using serial mirror sections. We found 15(6.3%) vimentin-positive cases. Twelve of them demonstrated varying amounts of rhabdoid-like cells. Eight cases diffusely co-expressed vimentin and cytokeratin simultaneously. In addition, four co-expressing cases showed positive staining with Keratin-903 which recognizes the high molecular-weight cytokeratin. Most of the co-expressing cases showed a diffuse proliferation of polygonal tumour cells with focal cell-to-cell contact. The prognosis of the co-expressing cases was poor in comparison with that of the 89 vimentin-negative tumours (P < 0.05).

Adenocarcinoma↗

PubMed1996

2-Deoxy-2-fluoro-D-glucose as a functional probe for NMR: the unique metabolism beyond its 6-phosphate.

Epimeric conversion of 2-deoxy-2-fluoro-D-glucose (FDG) to its 2-epimer 2-deoxy-2-fluoro-D-mannose (FDM) proved by 19F NMR has been shown to reflect the brain activity. To examine the feasibility of FDG as a new NMR probe for in vivo functional monitoring, we studied here the fundamental NMR properties of metabolites, spectral assignments, and reliability of NMR quantification. Metabolites confirmed in brain besides FDM-6-phosphate were as follows: FDG-1-phosphate, FDG-1,6-bisphosphate, FDM-1-phosphate, FDM-1,6-bisphosphate, and FDG and FDM derivatives of nucleotide diphosphate. NMR quantification of these metabolites was evaluated in comparison with the method of 18F-labeled FDG. In the NMR functional study using FDG, where a high dose is inevitable, the dose dependence of uptake was investigated. FDG uptake in mouse brain was shown to be in the range of interpretation using the biochemical parameters of enzymes for glucose uptake as long as a dose of < 200 mg/kg was used.

Animals↗

PubMed1996

Purification and refolding of recombinant human proMMP-7 (pro-matrilysin) expressed in Escherichia coli and its characterization.

Human matrix metalloproteinase-7 (MMP-7 = matrilysin) was overproduced in Escherichia coli as a recombinant zymogen (31 kDa), the C-terminus of which bears artificial hexa-histidines. Most of the enzyme was isolated from the insoluble fraction of the cell lysate and purified by a single step using Ni-NTA resin after solubilization of the precipitates with 8 M urea solution. The resin-bound recombinant protein was refolded into a form that is activatable by p-amino-phenylmercuric acetate in an autocatalytic manner. The activated enzyme cleaved a synthetic peptide substrate at the reported site for MMP-7. Digestion of carboxymethylated transferrin (a natural substrate of MMP-7) by the recombinant proteinase generated fragments with the same peptide map as in the case of native purified MMP-7. The autocatalytic activation and enzyme reaction were entirely dependent on the presence of calcium and zinc ions. The enzyme activity to cleave carboxymethylated transferrin was inhibited by tissue inhibitors of metalloproteinases-1 and -2, MMP-specific inhibitors. The activity of the recombinant MMP-7 was also inhibited by a synthetic peptide derived from a part of the cysteine switch that maintains the zymogen in an inactive state. Thus, we report here a simple means of preparing a large quantity of recombinant proMMP-7 that can be used to study the activation mechanism and to screen synthetic inhibitors.

Amino Acid Sequence↗

PubMed1996

Structure and expression of two seed-specific cDNA clones encoding stearoyl-acyl carrier protein desaturase from sesame, Sesamum indicum L.

We have isolated two cDNA clones (CDES01) and 04) encoding stearoyl-acyl carrier protein desaturase (SACPD; EC 1.14.99.6) from immature sesame seeds, and have analyzed accumulation levels of the corresponding mRNAs at different stages and organs in sesame. Clone CDES01 contains an open reading frame coding for a 396-amino acid protein of 45 kDa. CDES04 encodes a partial sequence of 141-amino acids. Deduced amino acid sequences of both clones exhibit a high identity to those of other plant SACPD cDNAs. Northern blots probed with CDES01 and CDES04 indicate that both messages accumulate in a seed-specific manner with a peak at 21 days after anthesis. However, expression patterns also indicate that regulation between CDES01 and CDES04 are slightly different. The CDES01 message accumulates at a low level in young leaves in addition to seeds, whereas accumulation of the RNA transcript corresponding to CDES04 is restricted to seeds. This observation implies the presence of at least two isozymes of SACPD having overlapping but slightly distinct functions in sesame.

Amino Acid Sequence↗

PubMed1996

Efficient production of the C-terminal domain of secretory leukoprotease inhibitor as a thrombin-cleavable fusion protein in Escherichia coli.

We have developed a high-level production system for the C-terminal domain of secretory leukoprotease inhibitor (SLPI) to investigate its pharmacological activities. A gene for the C-terminal domain of SLPI, (Asn55-Ala 107)SLPI, was constructed from chemically synthesized deoxyoligonucleotides. It was fused to a gene for the N-terminal portion of human growth hormone via a DNA sequence encoding Leu-Val-Pro-Arg, which can be cleaved by thrombin. The fused gene was expressed in Escherichia coli under the control of a trp promoter, and the fusion protein was obtained as an inclusion body. After sulfonation of the cysteine residues, the sulfonated fusion protein was cleaved at the desired site by thrombin. Sulfonated (Asn55-Ala107) SLPI was refolded in Tris buffer containing reduced and oxidized glutathione. The resulting (Asn55-Ala107) SLPI was purified by cation-exchange chromatography and reverse-phase high performance liquid chromatography. The final yield was 50 mg/I culture. (Asn55-Ala107) SLPI was as active against elastase as, but had less trypsin inhibitory activity than, native SLPI. This system is suitable for the large-scale production of the C-terminal domain of SLPI, which is an elastase-specific inhibitor.

Amino Acid Sequence↗

PubMed1996

Biological and antigenic characterization of three BApNA-hydrolyzing proteases from the culture supernatant of Porphyromonas gingivalis.

Biological and antigenic distinction of 3-N-alpha-benzoyl-DL-arginine p-nitroanilide (BApNA)-hydrolyzing proteases (Pase-B, Pase-C and Pase-S) isolated from the culture supernatant of Porphyromonas gingivalis were determined. Immunoblotting analysis of these enzymes using a polyclonal antibody against Pase-S, which is a soluble, clostripain-like protease, revealed immunological distinction from Pase-C, a vesicle-associated thiol-protease. Pase-B, a vesicle-associated clostripain-like protease, reacted with the antibody and was also found to contain a considerable amount of carbohydrates in its structure, as compared with the others. Analysis of N-terminal amino acids of Pase-B provided a sequence not found in the SwissProt data bank or previously reported as N-terminal sequences of proteases from P. gingivalis. Pase-S, resembling Pase-B in its hydrolytic specificity, cleaved only arginine residues of peptides and degraded type IV and denatured type I collagen. Pase-C hydrolyzed N-alpha-benzoyl-DL-lysine p-nitroanilide and showed the strongest capacity of degrading native type I collagen. This enzyme was also the only one to possess hemagglutinating activity. Our findings suggest that Pase-S from P. gingivalis is less active than Pase-C and that the enzyme may be an isozyme of Pase-B.

Affinity Labels↗

PubMed1996

Age-related occurrence of inhibitory antibodies to streptococcal pyrogenic superantigens.

Several bacteria, such as staphylococci and streptococci, can produce superantigens (SA) that induce the activation of T cells in humans. Although these organisms are the major causes of infection in children, the evidence that T cells are vigorously activated by SA produced by such organisms has not been reported except for toxic shock syndrome. In a previous paper, we demonstrated that inhibitory IgG antibodies (Ab) to SA in humans may protect against SA stimulation. In the present study, we investigated the occurrence of these inhibitory Ab to SA in 94 healthy children by the enzyme linked immunosorbent assay technique and the suppressive effect on T cell stimulation by SA. The positivity of Ab to streptococcal pyrogenic exotoxin (SPE)-A, SPE-C and staphylococcal enterotoxin B (SEB) increased with age. The age at which more than 50% of children exhibited Ab to SA was 1 year for SEB, 6 years for SPE-C and 11 years for SPE-A. Sera from these children were inhibitory to T cell proliferation elicited by SA in proportion to the concentration of IgG Ab to each SA. Sera supplemented with IgG Ab to SA by gamma-globulin therapy became inhibitory to T cell proliferation by SA. We conclude that, as children grow, they can develop Ab to SA that may play a role in protecting them against vigorous T cell activation by SA.

Adolescent↗

PubMed1996

Breast lesions: correlation of contrast medium enhancement patterns on MR images with histopathologic findings and tumor angiogenesis.

PURPOSE: To compare qualitative and quantitative magnetic resonance (MR) mammographic features of breast lesions with histopathologic findings, especially tumor angiogenesis. MATERIALS AND METHODS: Seventy-three patients (72 women, one man; aged 30-78 years; mean age, 51.0 years) with suspicious breast lesions underwent MR imaging. Noncontrast medium-enhanced localization imaging and then gadolinium-enhanced dynamic fast spoiled gradient-recalled-echo (SPGR) imaging were performed in all patients. In selected patients, subtraction fast SPGR images were obtained. The Pearson and Spearman correlation tests were used to determine the strength of the relationships between enhancement parameters and microvessel determinations. RESULTS: Time intensity curve type correlated with microvessel density grade (Spearman rank correlation test: r = .90, P < .001). The steepest slope of contrast medium uptake correlated with microvessel counts (Pearson correlation test; r = .83, P < .001). Peripheral enhancement in invasive carcinomas (n = 9) correlated with high peripheral and low central microvessel densities, which were associated with desmoplasia and/or necrosis. Internal septations (n = 2) were seen only in fibroadenomas. CONCLUSION: The density and distribution of microvessels may play major roles in the determination of the initial rate of contrast medium uptake and the heterogeneity of tumor enhancement.

Adult↗

PubMed1996

Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage.

The matrix formed by adult human chondrocytes in alginate beads is composed of two compartments: a thin rim of cell-associated matrix that corresponds to the pericellular and territorial matrix of articular cartilage and a more abundant further-removed matrix, the equivalent of the interterritorial matrix in the tissue. On day 30 of culture, the relative and absolute volumes occupied by the cells and each of the two matrix compartments in the beads were nearly identical to those in native articular cartilage. Furthermore, the concentration of aggrecan in the cell-associated matrix was similar to that in adult human articular cartilage and was approximately 40-fold higher than in the further removed matrix compartment. Fluorescence-activated cell sorting revealed that the cell-associated matrix was built on the cell membrane in part via interactions between hyaluronic acid and CD44-like receptors. Approximately 25% of the aggrecan molecules synthesized by the chondrocytes during a 4-h pulse in the presence of [35S]sulfate on day 9 of culture were retained in the cell-associated matrix where they turned over with a half-life (t1/2) = 29 days. Most [35S]aggrecan molecules reached the further removed matrix compartment where they turned over much more slowly (t1/2 > 100 days). These results add support to the contention that aggrecan molecules residing in the pericellular and territorial areas of the adult human articular cartilage matrix are more susceptible to degradation by proteolytic enzymes synthesized by the chondrocytes than those that inhabit the interterritorial areas further removed from the cells.

Adult↗

PubMed1996

Expression of hybrid isomyosins in human skeletal muscle.

Myosin of human skeletal muscles was analyzed by means of several electrophoretic techniques. Myosin heavy chain (HC)-IIa-and HC-IIb-based isomyosins were identified by pyrophosphate-polyacrylamide gel electrophoresis (PP-PAGE). The electrophoretic mobilities of these fast-twitch muscle isomyosins differed in the order HC-IIa triplets < HC-IIb triplets. To determine the subunit composition of myosin molecules that function in intact muscle, two-dimensional electrophoresis in which the first and second dimensions were PP-PAGE and sodium dodecyl sulfate-PAGE, respectively, was also performed. Slow-twitch muscle isomyosin contained, in addition to slow-twitch light chain (LC) and HC-I isoforms, appreciable amounts of LC-2f, HC-IIa, and HC-IIb isoforms, and fast-twitch muscle isomyosin consisted of LC-2s and HC-I isoforms as well as fast-twitch LC and HC isoforms. Without consideration of HC- and slow-twitch alkali LC heterodimers, at least 31 possible isomyosins are derived from these findings on the subunit composition of isomyosins in human skeletal muscle.

Diphosphates↗