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Biomedical subjects

T Matsuoka

Publications and source records attributed to T Matsuoka.

At least 19 recordsLinked to original sources

Fetal liver T cell receptor gamma/delta+ T cells as cytotoxic T lymphocytes specific for maternal alloantigens.

We have established fetal liver-derived T cell receptor (TCR) gamma/delta+, CD3+ T cell lines that are cytotoxic for maternal T cells. Fetal liver-derived lymphoid progenitors yielded predominantly TCR-gamma/delta+ cell clusters when cultured on fetal bone marrow-derived stromal cells in the presence of a cytokine cocktail under magnetic force. These tightly adherent clusters were cloned by limiting dilution and the resulting cell lines analyzed for phenotype and function. Six of eight TCR-gamma/delta lines from 8-9.5-wk gestation fetuses were V delta 2+ as compared with zero of eight lines from later stages of gestation (10 and 15 wk), where all the lines were V delta 1+. In cytotoxicity assays, these TCR-gamma/delta+, CD3+, CD4-, and CD8+ or CD8- long-term cultured lymphoid cells (LLC) were killer cells active against the class I antigens on maternal T cells. Of the cell lines, the CD8+ TCR-gamma/delta+ LLC had the highest levels of killer activity. Thus fetal liver TCR-gamma/delta+ T cells may play a crucial role in protection against invading maternal T cells generated in the feto-maternal interaction.

Cells, Cultured

Transcriptional regulation of the c-jun gene by retinoic acid and E1A during differentiation of F9 cells.

Differentiation of mouse F9 embryonal carcinoma (EC) cells can be induced by exposure to retinoic acid (RA) or by expression of adenovirus E1A. The transcription of the c-jun gene is stimulated by either RA or E1A. We report here that both RA and E1A strongly induce the expression of chloramphenicol acetyltransferase (CAT) from c-jun promoter/CAT reporter construct (c-jun/CAT), which is stably integrated into F9 cells, in a manner that is independent of both copy number and integration locus. The induction of c-jun/CAT expression is observed in undifferentiated F9 cells, but not in differentiated F9 cells, adenovirus-infected F9 cells or HeLa cells. Deletion analysis of the promoter region of the c-jun gene indicates that the sequence elements required for the RA- and E1A-mediated induction are identical and they have been defined as a region of 145 bp between -190 and -46 of the 5' flanking region of c-jun. This RA and E1A response element (RERE) contains five variants of the motif CGCGGTGACGNT. The upstream two motifs are adjacent and extend in opposite directions, creating an imperfect palindrome. The downstream four motifs are located at 35 or 36 bp intervals in the same orientation. Substitution and insertion analysis indicates that these motifs and their regular intervals are important for the activity of the RERE.

Adenoviruses, Human

Segmental cytochrome c-oxidase deficiency in CPEO: teased muscle fiber analysis.

In an attempt to elucidate the pathogenesis of focal cytochrome c-oxidase (COX) deficiency in skeletal muscle from patients with chronic progressive external ophthalmoplegia (CPEO), we examined the longitudinal distribution of COX activity in single muscle fibers from 6 CPEO patients with muscle mitochondrial DNA (mtDNA) deletions. A new method for teasing single muscle fibers, recently developed in our laboratory, revealed fibers with COX-positive and -negative segments in all 6 patients. The borders between the enzyme-positive and -negative segments in these fibers were sharply delineated, so that the length of each COX-negative segments could be accurately measured. The proportion of the sum of the lengths of the enzyme-negative segments to the total length of the muscle fibers correlated well with the proportion of deleted mtDNA, suggesting that abnormal mitochondria harboring mutant mtDNA may be responsible for the focal loss of COX activity.

Adolescent

An experimental model of mitochondrial myopathy: germanium-induced myopathy and coenzyme Q10 administration.

In skeletal muscles from rats treated with germanium for 23 weeks, there were numerous ragged-red fibers and cytochrome-c oxidase (COX)-deficient fibers. Biochemically, germanium reduced the enzyme activities in the mitochondrial respiratory chain. Rotenone-sensitive NADH-cytochrome-c reductase as well as COX activities were markedly reduced, while succinate-cytochrome-c reductase was less severely, but significantly, affected. The histopathological findings in these muscles were similar to those seen in patients with mitochondrial encephalomyopathy, suggesting that germanium-induced myopathy may be a useful experimental model. Coenzyme Q10 administration appeared to be ineffective in preventing this experimental myopathy.

Animals

The topographical features of EEGs in patients with affective disorders.

EEG data were obtained in the basic state from 16 scalp sites of 44 patients with affective disorder, diagnosed by DSM-III criteria, and 44 normal controls. The EEG power spectra were computed and the t statistic significance probability mapping (SPM) was applied to visualize regions where the patient group showed differences in the EEG topogram from the controls. The results show: (1) left occipital predominance (P3, O1) of alpha activities in the patients with affective disorders, (2) decreased alpha activities in Fp2 and F8 areas in patients with major depression without melancholia, (3) decreased alpha activities in F7 area in patients with bipolar disorder, manic, and (4) increased beta 2 activity in F4 and C4 areas in patients with major depression with melancholia. These results suggest that inter-hemispheric and intra-hemispheric relationships may be disturbed in patients with affective disorder.

Adult

Fractal dimension of dendritic tree of cerebellar Purkinje cell during onto- and phylogenetic development.

The cerebellar Purkinje cell has highly branched dendrites extending in a transverse plane to the cerebellar folium. Branching patterns of the dendrites were subjected to fractal analysis to obtain a quantitative measure of their intricacy. Using a square-covering method, we found that dendrites have a statistical self-similarity so that their fractal dimension can be evaluated. In postnatal mouse, dimensions of Golgi-stained Purkinje cells increased as dendrites developed. The time course of the dimension increase was correlated with development of the width and area of the dendritic fields. After the dimension reached the maximum value at postnatal day 20, the width and area further continued to grow until day 100. In human cerebellum, dimensions of Purkinje cell dendrites increased greatly during pregnancy and reached 97% of the adult value at birth. On the other hand, 41% of the adult dendritic field width and 11% of the adult area had been attained at birth. The physical size of dendrites increased in a more lagged phase than the dimensions, and growth proceeded mainly after birth. These observations indicate that after the maximum complexity and fundamental tree pattern have been attained, the overall tree size further increases. It is observed that phylogenetic development of dendritic trees of Purkinje cells in several species of vertebrates showed a correlation with the increase in dimensions. These studies demonstrated the potency of the fractal analysis in evaluating development of dendritic trees of neurons.

Aging

Membrane potential and current responses to neurotensin in the longitudinal muscle of the rectum of the fowl.

1. The effects of neurotensin (NT) on membrane potential and membrane current of the longitudinal smooth muscle of chicken rectum were investigated by intracellular recording and whole-cell voltage clamp. 2. NT (3 nM-1.2 microM), when applied via the bathing medium, produced a concentration-dependent membrane depolarization with an EC50 of 18 +/- 2 nM (n = 7) which was accompanied by an increase in the membrane conductance. The effect was biphasic: an initial, rapid depolarization reached a peak within 2-3 min and then declined to a lower but still elevated level which was sustained until washout. 3. Excitatory junction potentials (e.j.ps), which were non-adrenergic non-cholinergic (NANC) in nature, were decreased in amplitude and total duration in the presence of NT (0.6 microM). The depression of the e.j.p. was due mainly to the reduction of the membrane resistance. 4. When NT was applied locally by means of pressure ejection from a micropipette containing NT, some cells responded with a membrane depolarization and some failed to respond, whereas e.j.ps could invariably be elicited from all of them. 5. In single muscle cells enzymatically isolated from the muscle and dialyzed under voltage clamp at -50 mV with a CsCl-rich solution, NT (5 or 10 microM) produced an inward current. NT-induced inward currents were obtained with inclusion of 10 mM EGTA in the pipette solution and their reversal potential was around 0 mV. In cells dialyzed under voltage clamp at 0 mV with a KCl-rich solution, NT (5 microM) produced a brief outward current followed by abolition of spontaneous transient outward currents.6. The present results suggest that the membrane depolarization, which may arise from activation of non-selective cation channels, and release of calcium from internal stores produced by neurotensin are responsible for its contractile activity in the longitudinal smooth muscle of chicken rectum. Further, the depolarizing effect may provide support for the involvement of NT in the NANC transmission in this preparation.

Adenosine Triphosphate

Identification of Pseudomonas aeruginosa by using a disk of phenanthroline and 9-chloro-9-[4-(diethylamino)phenyl]-9,10-dihydro-10-phenylacridine hydrochloride and by cell agglutination testing with monoclonal antibodies.

To establish a simple identification procedure for Pseudomonas aeruginosa, we developed a disk consisting of phenanthroline and 9-chloro-9-[4-(diethylamino)phenyl]-9,10-dihydro-10- phenylacridine hydrochloride (PC disk). Nine of 248 P. aeruginosa isolates and all other oxidase-positive gram-negative isolates (143 isolates) had clear inhibition zones around the PC disk. Of these nine P. aeruginosa isolates, seven produced a blue, blue-green, or green pigment on Mueller-Hinton agar after 24 h of incubation. The sensitivity and specificity of the PC disk susceptibility test in combination with pyocyanin production were 99 and 100%, respectively. Ten P. aeruginosa isolates showed no agglutination reactions with monoclonal antibodies against P. aeruginosa (96% sensitivity and 100% specificity).

Acridines

Ventilatory and metabolic responses to acute hyperoxia in newborns.

Hyperoxia has previously been found to increase metabolic rate (oxygen consumption [VO2] and CO2 production [VCO2]) in newborn mammals. We asked whether the same occurs in the newborn infant. Breathing pattern was measured in 25 full-term infants, 1 to 2 days of age, from the spirometric record obtained with a pneumotachograph attached to a face mask. Concentrations of O2 and CO2 were continuously measured at the mouth; VO2 and VCO2 were computed as the product of VE and the difference between inspired and expired concentration of the respective gases, 5 min of air (FIO2 = 0.21) and 5 min of O2 (FIO2 = 1). A bias flow through the mask and pneumotachograph delivered the inspired gas and eliminated the effects of the instrumental dead space. In neither case did measurements at 1 min significantly differ from those taken at 5 min. In hyperoxia VE increased in 22 of the 25 infants, in average +18% (p less than 0.001, paired two-tailed t test). Because of a rise in tidal volume (+35%, p less than 0.001) and a decrease in breathing rate (-11%, p less than 0.005) alveolar ventilation (VA) increased by about 58% (p less than 0.001). VO2 and VCO2 increased by 25% and 17%, respectively (p less than 0.001). The rise in VO2 was too large to be explained by the greater respiratory work of the hyperventilation, whereas that of VCO2 was not large enough to fully explain the increase in VA. We conclude that in newborn humans, as in other newborn species, the normoxic metabolic rate seems to be limited by the availability of O2.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Dioxide

Lung capillary endothelial cells produce and secrete urokinase-type plasminogen activator.

Bovine lung capillary endothelial cells (BLuEC) were isolated, and their ability to produce plasminogen activator (PA) in vitro was demonstrated. BLuEC secreted more than 10 times as much as urokinase-type PA (u-PA) as did bovine aortic, hepatic capillary and adrenal capillary endothelial cells, and lung fibroblasts. BLuEC secreted u-PA on both sides of the cell layer, the luminal surface, and the basic surface attached to the basement membrane. u-PA mRNA was detected in BLuEC by Northern blotting, but not in endothelial cells from other tissues and fibroblasts. These results suggest that BLuEC may contribute not only to the patency of lung vessels but also to the maintenance of alveolar functions through the production and secretion of u-PA.

Amino Acid Sequence

Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS): a correlative study of the clinical features and mitochondrial DNA mutation.

We studied 40 MELAS patients (21 male and 19 female) to characterize the clinical features and biochemical and muscle biopsy findings related to the mtDNA mutation at the nucleotide position of 3,243, the most common genetic defect in MELAS. The most frequent symptom was episodic sudden headache with vomiting and convulsions, which commonly affected patients aged 5 to 15 years (80%). Biochemical defects in the muscle were variable; 13 patients had complex I, seven complex IV, and four complexes I + IV deficiencies. In four muscle biopsies without ragged-red fibers or any enzyme defect, we based the diagnosis on the identification of strongly SDH-reactive blood vessels, which occurred in 87.5% of the biopsies. The mtDNA mutation was present in 32 of 40 patients (80%). We conclude that there are no clinical and pathologic differences between the patients with and without this mtDNA mutation.

Acidosis, Lactic

The role of the sodium ion in calcium-free solution on the mechanism of the calcium paradox in the guinea pig heart.

We examined the effect of low Na concentrations (25 mM) in Ca-free solutions on the myocardial damage induced by the Ca paradox in guinea pigs using electron microscopy and electrophysiological methods. Ultrastructural damage consist of contraction bands, swelling of mitochondria and disappearance of major part of contractile apparatus induced by the Ca paradox was prevented by reducing Na from 145 to 25 mM in the Ca-free solution. The penetration of lanthanum into the myocardial cells, which was induced by the Ca paradox, was also prevented by reducing Na concentration during the Ca depletion period. Perfusion with Ca-free solution containing normal Na (145 mM) for 10 min caused a significant prolongation in the action potential duration (APD) of left ventricular papillary muscle, and reperfusion with Ca containing solution caused depolarization. The perfusion with Ca-free solution containing low Na concentration (25 mM) significantly reduced APD, but APD returned almost to control values without depolarization after reperfusion with Ca-containing solution for 5 min. These results suggest that the Na concentration in a Ca-free solution plays an important role in the pathogenesis of the Ca paradox phenomenon in guinea pig hearts.

Action Potentials

[The reactivity of dispersed human lung mast cells and peripheral blood basophils to acetylcholine].

To clarify the relation between human lung mast cells and parasympathetic nerve function as well as IgE mediated allergic reactions, highly purified dispersed human lung mast cells were obtained by using the techniques of scissors dispersion, enzymatic treatment, percoll centrifugation and exclusion of adherent cells. The reactivity to acetylcholine was examined by observing the histamine release of purified mast cells. Moreover, peripheral blood basophils, which have many functional similarities with mast cells, were also examined in the same manner. The following results were obtained; 1) Histamine was significantly released from dispersed human lung mast cells at a final concentration of 10(-5) M acetylcholine (p less than 0.05); the peak of histamine release was 10(-4) M of acetylcholine. Acetylcholine had the additional effect of releasing histamine in response to anti-IgE. Histamine release was partially inhibited by 10(-5) M atropine. 2) Basophils had no response to acetylcholine. These results suggest that human lung mast cells play an important role in the defensive mechanism as an effector cell of acetylcholine-mediated autonomic nerve system as well as IgE-mediated allergic reaction.

Acetylcholine

[Strongly succinate dehydrogenase-reactive blood vessels (SSV) in various neuromuscular diseases].

The strongly succinate dehydrogenase-reactive blood vessels (SSV) are shown to have increased numbers of enlarged mitochondria in smooth muscle cells of the vessel wall on electron microscopy. They are seen in biopsied skeletal muscles from patients with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) at high frequency. The present study was done to examine the incidence of SSV in biopsied muscles from various neuromuscular diseases. Among 107 patients with mitochondrial encephalomyopathies (MEM) including 50 with chronic progressive external ophthalmoplegia (CPEO), 7 with myoclonus epilepsy with ragged-red fibers (MERRF), and 50 with MELAS, SSV were seen in nearly a half of the patients, and comprised approximately 24% of small arteries. On the other hand, SSV in 100 patients with various neuromuscular diseases other than MEM were exceptional, and only one of 8 patients with myotonic dystrophy had SSV. These findings suggest that the SSV are induced by functional abnormality of mitochondria in smooth muscle cells, and that an identification of the SSV is an additional crucial evidence to make a pathological diagnosis of MEM.

Blood Vessels