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Biomedical subjects

T Tsuda

Publications and source records attributed to T Tsuda.

At least 145 records · Page 8Linked to original sources

A clearance model of inhaled man-made fibers in rat lungs.

A clearance model of inhaled man-made fibers (MMFs) was developed, and the calculated fiber numbers and dimensions were compared with the experimental ones using a glass fiber (GF), ceramic fiber (RF1) and two potassium octatitanate whiskers (PT1, TW). If the translocation rate by macrophages is constant and the effect of dissolution and disintegration can be ignored, the fiber number is expected to decrease exponentially with time. In the experimental study, however, the fiber number did not always decrease exponentially. In the case of RF1, the fiber number decreased almost exponentially and the diameter decreased linearly with the time. The clearance rate constant of GF during 3 to 6 months after the end of one-month exposure was greater than that during 1 to 3 months. On the contrary, the clearance rate constants of PT1 and TW during 1 to 6 months were greater than next six months. The diameter and the length of GF did not change significantly. The fiber length of PT1 tends to become longer with time although the diameter did not change significantly. Our theoretical model gives a satisfactory fit to these experimental results.

Administration, Inhalation↗

Effects of mineral fibers on the gene expression of proinflammatory cytokines and inducible nitric-oxide synthase in alveolar macrophages.

To determine which parameters are useful for the risk assessment of man-made mineral fibers (MMMFs), we examined the gene expression of interleukin-1 alpha (IL-1 alpha), tumor necrosis factor alpha (TNF alpha), interleukin-6 (IL-6) and inducible nitric-oxide synthase (iNOS) in mineral fiber-exposed alveolar macrophages (AMs). Male Wistar rats were intratracheally exposed to saline or mineral fibers suspended in saline (2 mg of crocidolite, chrysotile, alumina silicate refractory fiber (RF1) or potassium octatitanate whisker (TW)). Bronchoalveolar lavage was performed 4 weeks after the fiber-instillation, and the recovered AMs were stimulated by lipopolysaccharide for 2 or 6 hours. Expression of IL-1 alpha, TNF alpha, IL-6 and iNOS from AMs was observed using reverse transcription-polymerase chain reaction (RT-PCR). The levels of IL-1 alpha and IL-6 mRNA induced by mineral fiber exposure were greatest in AMs exposed to TW, crocidolite, chrysotile and RF1 in that order. However, both gene expression of iNOS and TNF alpha were not elevated in both crocidolite and TW exposure, despite their high pathological potential. These data suggested that IL-1 alpha and IL-6 may be useful indicators for the risk assessment of MMMFs.

Administration, Inhalation↗

[Acute empyema caused by Streptococcus milleri in early pregnancy].

A 40-year-old woman was admitted to our department complaining of left lateral chest pain and fever. She smoked 10 cigarettes per day. Chest radiographs revealed increased density in the upper left lung field and air-fluid level in the lower left lung field. A diagnosis of acute empyema was made, because pus was aspirated by thoracentesis. Streptococcus intermedius (Streptococcus milleri group) was isolated from samples of pleural effusion. The patient was successfully treated with a combination of antibiotics and surgical drainage. It became clear after discharge that she was in her 6th week of pregnancy. Laboratory findings showed decreased lymphocyte transformation in the PHA and Con-A tests. We reasoned, therefore, that smoking and decreased cellular immunity due to pregnancy might be causes of bacterial infections such as empyema.

Acute Disease↗

[Two adult siblings with Chédiak-Higashi syndrome presenting as hyperpigmentation of the skin and the iris].

We report two adult siblings with Chédiak-Higashi syndrome presenting as hyperpigmentation of the skin and the iris. Patient 1 was a 30-year-old man who had generalized hyperpigmentation from one month of age, developed mental deterioration at age 9 years, and gait difficulty at age 20 years. On admission, he showed hyperpigmentation of the skin and the iris without partial albinism. Skin pigmentation was predominated at the face and the extremities. Neurologic examinations revealed mental dysfunction with IQ of 60 by WAIS, cerebellar ataxia, pyramidal signs, extrapyramidal signs, and polyneuropathy. Hematologic examination revealed peroxidase-positive giant granules in leukocytes and decreased activity of natural killer cells, leading to the diagnosis of Chédiak-Higashi syndrome. Patient 2, a younger brother of patient 1, was a 28 year-old man who also had hyperpigmentation of the skin and the iris with similar neurologic findings with patient 1. Both had no episodes of systemic infections. In Japanese cases of Chédiak-Higashi syndrome, more than 50% of them showed hyperpigmentation of the skin from the early stage of the disease. We pointed out that hyperpigmentation of the skin may be a good diagnostic help of Chédiak-Higashi syndrome in Japanese cases.

Adult↗

Regulation of gene expression by herbal medicines--a new paradigm of gene therapy for multifocal abnormalities of genes.

Japanese herbal medicine has long been considered as only supplementary therapy to Western medicine. However, we discovered that an herbal mixture, Saiko-keishi-to-ka shakuyaku (SK, TJ-960), showed regulatory function of gene expression such as increased expression of seizure-related gene PTZ-17, proto-oncogene c-fos and heat shock protein HSP 72. These results provide a scientific basis for an important ancient concept and usage of herbal mixtures as a "therapy against diseases which will be suffered in the future". Our results also give an adequate provide break-throughs for therapy and even prevention of intractable epilepsy, Alzheimer's disease, developmental disorders during pregnancy and the postnatal period, and also probably for prevention of metastasis or relapse of various cancers.

Animals↗

Biodegradation and tumorigenicity of implanted plates made from a copolymer of epsilon-caprolactone and L-lactide in rat.

Flat plates made from a copolymer of epsilon-caprolactone and L-lactide (P-CL-LA) [50:50 (w/w), molecular weight 1.62 x 10(5); 20 x 10 x 1 mm size] were subcutaneously implanted into 50 young, male Wistar rats (P-CL-LA group). After 24 months the plates had become a mass of small pieces, which were concentrated in an area of 3 x 2 x 1 mm. For comparison, 50 rats were implanted with medical-grade polyethylene plates (PE group) while another set of 50 rats was subjected to the same operation but without an implant (Sham Op group). Tumors arose in 25 rats from the P-CL-LA group: 24 were malignant mesenchymal tumors at the implant sites. In the PE group, tumors appeared in 16 rats (14 at the implant sites and two ectopically). The average tumor latency was 578+/-84 days in the P-CL-LA group and 452+/-102 days in the PE group. There was no difference in tumor incidence between the P-CL-LA and PE groups (p < 0.05). In the Sham Op group, two malignant tumors appeared over 2 years. Pathologically, these induced tumors arose from the inflammatory cells surrounding the degrading fragments of P-CL-LA within the tissue capsule. This indicates that relatively slowly degrading material can induce malignant tumors at a similarly high rate to nonabsorbable medical grade PE, at least in this animal model.

Abdominal Neoplasms↗

Expressed emotion of families and negative/depressive symptoms in schizophrenia: a cohort study in Japan.

This study investigated whether the Expressed Emotion (EE) status of families is associated with an increased risk of negative and depressive symptoms in schizophrenia. The subjects were 52 schizophrenic patients from mental hospitals in Kochi, Japan, who satisfied the DSM-III-R or ICD-9 criteria for schizophrenia. The Japanese version of the Camberwell Family Interview was administered to 73 key relatives of the patients within 2 weeks after admission. A certified rater evaluated the EE's status of each family, using an audiotaped interview and its transcript. Using cohort study design, the subjects were followed for 9 months after their discharge and were monitored for negative and depressive symptoms. Trained psychiatrists who were blind to the EE status of the patients' households administered the Brief Psychiatric Rating scale (BPRS) at discharge, and 9 months after discharge. Repeated-measures analyses of variance showed that EE effects were significant in Emotional withdrawal in all subjects, Depressive mood and Total depressive symptoms in non-relapsers. A multiple regression analyses revealed that EE's effect was significant only in Depressive mood in non-relapsers controlling confounding factors. In non-relapsers in positive symptoms, high EE families could make patients depressed. EE's effect on negative symptoms remains to be established.

Adolescent↗

Modeling and optimization of enantioseparation by capillary electrochromatography.

Both electrophoretic and chromatographic transport mechanisms are combined in electrochromatographic separation. In this paper, we developed a model of enantioselectivity in capillary electrochromatography (CEC) which can be applied in the separation of both neutral and ionic compounds. The overall selectivity in enantioseparation is considered to be made up of two contributions: one is the intrinsic difference in formation constants of a pair of enantiomers, and the other is the conversion efficiency of the intrinsic difference into the apparent difference in the migration velocity. The model was illustrated through the chiral separation of (R)- and (S)-salsolinols. Under a positive electric field, enantioseparation of salsolinols was achieved on an ODS column with beta-cyclodextrin as a chiral mobile-phase additive. The experimental results are discussed in relation to the effect of separation parameters, such as the direction and size of electric field and properties of the stationary and mobile phases. It was demonstrated that if both electrophoretic and partitioning mechanisms produce positive effects, high overall selectivity in CEC can be obtained. For pressurized-driven electrochromatography, although the column efficiency is sacrificed due to the introduction of hydrodynamic flow, the increased selectivity significantly reduced the requirement of large column plate numbers for resolution.

Buffers↗

Half-site modification of Lys-480 of the Na+,K+-ATPase alpha-chain with pyridoxal 5'-diphospho-5'-adenosine reduces ATP-dependent phosphorylation stoichiometry from half to a quarter.

Pig and dog kidney Na+,K+-ATPase preparations, irrespective of specific activity, showed approximately 0.5 mol of maximum phosphorylation/mol alpha-chain for ATP or acetyl phosphate (AcP) at steady state conditions. Pyridoxal 5'-diphospho-5'-adenosine (AP2PL)-treated pig kidney enzymes containing approximately 0.5 mol of AP2PL probe at Lys-480/mol (Tsuda, T., Kaya, S., Funatsu, H., Hayashi, Y., and Taniguchi, K. (1998) J. Biochem. (Tokyo) 123, 169-174) showed a quarter-site phosphorylation by ATP and half-site phosphorylation from AcP. The addition of 10 microM ATP to the Mg2+-Na+-bound AP2PL enzyme induced rapid quarter-site phosphorylation (47/s), followed by two different AP2PL fluorescence changes, a rapid decrease (29/s) and a slow increase (1.1/s). The addition of 1 mM AcP to the Mg2+-Na+-bound AP2PL enzyme induced a slow half-site phosphorylation (3/s), followed by a monophasic AP2PL fluorescence increase (1.2/s). After treatment of the AP2PL enzyme with fluorescein 5'-isothiocyanate to modify Lys-501 fully, the Mg2+-Na+-dependent phosphorylation capacity from ATP of the resulting AP2PL-fluorescein 5'-isothiocyanate enzyme was reduced to approximately 6% without significant changes in half-site phosphorylation capacity with respect to AcP, dynamic AP2PL fluorescence change by ATP and change by AcP. These data and others support the hypothesis that the functional membrane-bound Na+, K+-ATPase has tetrameric properties.

Adenosine Diphosphate↗

ATP and acetyl phosphate induces molecular events near the ATP binding site and the membrane domain of Na+,K+-ATPase. The tetrameric nature of the enzyme.

The addition of ATP to Mg2+-Na+-bound-probe labeled Na+,K+-ATPase preparations containing approximately 0.5 mol of pyridoxal 5'-diphospho-5'-adenosine (AP2PL) probe at Lys-480 and approximately 0.9 mol of fluorescein 5'-isothiocyanate (FITC) probe at Lys-501 showed a decrease and an increase in the AP2PL fluorescence intensity with neither significant ATP-dependent phosphorylation nor FITC fluorescence change. The rate constants for the fluorescence change increased nearly linearly with increasing ATP concentrations. The substitution of AcP for ATP decreased the FITC fluorescence rather monophasically, 8.5/s, which was followed by the half-site phosphorylation with same amount of components with different rate constant, 7.2 and 4.6/s, followed by a much slower increase in the two components of AP2PL fluorescence, 1.4 and 0.2/s. The addition of Na+ with increasing concentrations of ATP to the K+-bound AP2PL-FITC enzymes induced accelerations in the decrease and an increase in the AP2PL fluorescence intensity with two different increases in the FITC fluorescence intensity, showing that the same concentration of ATP is capable of inducing four different fluorescence changes. The addition of ATP to the Mg2+-Na+-bound enzymes modified with N-[p-(2-benzimidazolyl)phenyl]-maleimide (BIPM) at Cys-964 and retaining full Na+,K+-ATPase activity induced two different increases in BIPM fluorescence intensity. Each rate constant for the BIPM fluorescence change versus concentrations of ATP gave two intersecting straight lines. These data and the stoichiometries of fluorescence probe bindings and ATP- and AcP-dependent phosphorylation provide strong support for the conclusion that the functional membrane-bound Na+,K+-ATPase is a tetramer.

Adenosine Diphosphate↗

Neuromedin B activates phospholipase D through both PKC-dependent and PKC-independent mechanisms.

The actions of neuromedin B (NMB), a recently discovered mammalian bombesin-related peptide, are mediated by interacting with a distinct receptor; however, little is known about its cellular basis of action. Recent studies show activation of phospholipase D (PLD) is an important transduction cascade for a number of GI hormones, especially for stimulation of growth and protein sorting. The purpose of the present study was to determine whether activation of the NMB receptor causes activation of PLD and to explore whether this activation was coupled to PLC activation. Rat C6 glioblastoma cells (C6 cells), which contain a low density of native NMB receptors and BALB 3T3 cells stably transfected with rat NMB receptors, were used. NMB caused a 3-fold increase in C6 cells and an 11-fold increase in rNMB-R transfected cells in PLD activity. Increases in PLD activity were rapid and NMB was 100-fold more potent than gastrin-releasing peptide (GRP). NMB caused a half-maximal increase in [Ca2+]i at 0.2 nM, in [3H]IP and PLD at 1 nM, and half-maximal receptor occupation at 1.2 nM. TPA increased PLD dose-dependently with a half-maximal effect at 60 nM. The calcium ionophore A23187 (1 microM) alone did not increase PLD activity but potentiated the effect of TPA. The Ca2+-ATPase inhibitor, thapsigargin, did not affect NMB- or TPA-stimulated PLD activities, although it blocked completely the NMB-induced increase in [Ca2+]i. The PKC inhibitor GF109203X completely abolished TPA-induced PLD activity, however, it only inhibited NMB-induced PLD activity by 20%. The combination of thapsigargin and GF109203X had the same effect as GF109203X alone. These data indicate that NMB receptor activation is coupled to both PLC and PLD. In contrast to a number of other phospholipase C-coupled receptors, NMB receptor stimulated changes in [Ca2+]i do not contribute to PLD activation. Both PKC-dependent and PKC-independent mechanisms are involved in the NMB-stimulated PLD activation with the PKC-independent pathway predominating.

3T3 Cells↗

Expression of three calpain isoform genes in human skeletal muscles.

Calpain is thought to be involved in muscular degradation in progressive muscular dystrophy (PMD), especially Duchenne and Becker muscular dystrophies. To assess the expression of calpain genes in skeletal muscles of patients with myopathies, we examined mRNA levels of three calpain isoforms by the quantitative reverse transcriptase-polymerase chain reaction method in biopsied muscles from control, PMD and amyotrophic lateral sclerosis (ALS) patients. There was a statistically significant increase in calpain 1 and calpain 2 mRNA levels in PMD and ALS patients as compared to controls. In contrast, there was a decrease in expression of calpain 3 mRNA in PMD, but it was not statistically significant. Expression of calpain 1 and calpain 2 positively correlated with each other, but not with calpain 3. These results indicate that expression of calpain 1 and calpain 2, but not calpain 3, are upregulated in diseased human muscles, likely playing a regulatory role in the process of myofibrillar degradation at the transcriptional as well as posttranslational level.

Adolescent↗

Localized muscle wasting as an initial symptom of skeletal muscle lymphoma.

A 25-year-old man who developed painless muscle wasting and weakness of only the right thigh, was eventually shown to have systemic lymphoma with muscle infiltration. The patient was initially misdiagnosed as focal inflammatory myopathy, and he had a partial response to steroid therapy. Immunohistochemical analysis of the biopsied muscle and the subcutaneous tumor revealed the infiltrating cells with a positive macrophage-associated marker (CD68) and negative T- or B-cell-associated antigens, leading to the final diagnosis of true histiocytic lymphoma. Although skeletal muscle lymphoma is extremely rare, it should be considered in the differential diagnosis of localized muscle wasting.

Adult↗

Effect of chloroquine-induced myopathy on rat soleus muscle sarcoplasm and expression of clathrin.

Clathrin-coated vesicles are involved in receptor-mediated intracellular transport pathways related to lysosomal proteolysis. Clathrin levels were significantly elevated in denervated soleus muscles from chloroquine- and saline-treated rats as compared with their contralateral, innervated muscles. No difference was found in the clathrin levels of the denervated muscles in both groups. The accumulation of autophagic vacuoles was marked only in chloroquine-treated muscles after denervation. These findings suggest that chloroquine does not inhibit intracellular trafficking of clathrin-coated vesicles during the overdevelopment of autophagic vacuoles.

Animals↗

Differential expression of flectin in the extracellular matrix and left-right asymmetry in mouse embryonic heart during looping stages.

A novel extracellular matrix protein flectin (250 kD M(r)) shows specific left-right asymmetric expression before and throughout the looping process during heart development in avian embryos [Tsuda et al., 1996]. Flectin is a candidate molecule to provide directionality to the looping process in the avian model. In this study on mouse embryonic heart development, flectin is shown to be developmentally regulated and to be expressed in a specific asymmetric fashion, but in a different pattern from that observed in avian hearts. The molecules involved in development tend to be the same, but timing of expression, modulation, and asymmetry are different. In the mouse embryo, flectin is expressed symmetrically when the cardiogenic plate is formed. As looping progresses, flectin expression becomes asymmetric. There is right side predominance at the outflow tract and left side predominance at the ventricular portion of the tubular heart. The left side predominance of flectin develops in an anteroposterior direction, while right side predominance of the outflow tract remains relatively unchanged. These differential expression patterns of flectin decrease once the looping process is completed. After looping, flectin becomes restricted to the epicardium and subepicardial extracellular regions. In inv/inv mice, a known mouse model for human situs inversus, in which the directionality of heart looping is inverted, flectin expression pattern is mirror image of that of normal mouse embryos during looping stages. Our study indicates that, in the mouse, flectin shows a specific asymmetric expression pattern after initiation of heart looping and that this asymmetric expression pattern is related to the directionality of looping. The remodeling of the extracellular matrix (ECM) including specific flectin expression begins with the looping process. This morphogenetic change of the ECM coincides with the differentiation of each region of the tubular heart.

Animals↗

[Removal of retained transvenous pacemaker electrodes under cardiopulmonary bypass].

A case of breakage and removal of a retained transvenous pacemaker electrode is described. A 22-year-old woman with complete A-V block underwent implantation of a transvenous pacemaker system on the left anterior chest wall in 1989. Three years later, a new generator was implanted on the right chest wall because of local infection of the pacemaker pockets. The old electrodes could not be removed and were left in place. Beginning in 1995, the patient complained of anterior chest pain. A chest roentgenogram revealed that one of the pacemaker electrodes had broken at the right costoclavicular ligament and a fragment was floating in the superior vena vava. The retained electrodes were removed under tot cardiopulmonary bypass. These electrodes had become firmly encased with fibrous tissue within the right ventricle and atrium, but they were easily removed under direct vision duting complete cardiac arrest. The postoperative course was uneventful and the patients had no further complaint.

Adult↗