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Biomedical subjects

T Kono

Publications and source records attributed to T Kono.

At least 163 records · Page 9Linked to original sources

Nuclei from fertilized mouse embryos have calcium-releasing activity.

During mammalian fertilization, the sperm triggers a series of intracellular Ca2+ oscillations which initiate oocyte activation and the formation of pronuclei. Oocyte activation can be induced artificially by a variety of chemical and physical stimuli which elevate intracellular calcium. We show that the transfer of nuclei from 1- and 2-cell-stage fertilized mouse embryos to unfertilized oocytes stimulates the completion of meiosis and the formation of pronuclei. Nuclei from embryos that had developed to the 4-cell stage did not stimulate meiotic resumption. The ability to cause oocyte activation was specific to nuclei transferred from fertilized embryos as nuclei from parthenogenetic embryos or cytoplasts from fertilized or parthenogenetic embryos did not induce activation. Nucleus-induced oocyte activation was associated with the generation of intracellular Ca2+ transients, which were seen after nuclear envelope breakdown of the transferred nuclei. Treatment of the oocyte with the intracellular Ca2+ chelator, BAPTA, prior to nuclear transfer inhibited intracellular Ca2+ transients and oocyte activation. The specific Ca(2+)-releasing activity of the nucleus was not caused by sperm-induced protein synthesis since similar activity was present in nuclei originating from embryos exposed to cycloheximide throughout fertilization. The specific ability of nuclei from fertilized embryos to stimulate Ca2+ transients and oocyte activation was also found in nuclei from embryos parthenogenetically activated by the injection of a partially purified cytosolic sperm factor. The results suggest that the fertilizing sperm introduces Ca(2+)-releasing activity which becomes associated with the nucleus of early mammalian embryos.

Animals↗

Functional characterization and localization of a cardiac-type inwardly rectifying K+ channel.

We cloned inwardly rectifying K+ channel cDNAs from porcine, rat and human, which were structurally almost identical with recently reported CIR(cKATP-1). The expression of CIR alone was low and unstable in Xenopus oocytes. The CIR/GIRK1 co-expression showed an increased current amplitude. Both the CIR and CIR/GIRK1 currents increased by coexpressing G beta gamma. The CIR and CIR/GIRK1 currents displayed two qualitative differences. (1) The conductance of the CIR channel did not saturate, but that of the CIR/GIRK1 channel showed saturation at hyperpolarized potential. (2) The CIR current showed instantaneous activation upon hyperpolarization, whereas the CIR/GIRK1 current exhibited slow activation, which was fitted by the sum of two exponentials. The CIR/GIRK1 current was also different from the GIRK1 current. The activation of the CIR/GIRK1 current was approximately ten times faster than that of the GIRK1 current. The increase in current amplitude and the qualitative differences imply the formation of functional heteromultimer. The CIR/GIRK1 channel showed differences from the native muscarinic K+ channel in that the basal level before m2 receptor activation is significantly large, and that the activation kinetics are much faster. Using anti-CIR antiserum, the CIR was detected in myocardial cells of the atrium and the ventricular subendocardial layer, and in the cardiac ganglion.

Amino Acid Sequence↗

RNA quadruplex containing G and A.

The CD study of r(GGAGGAA) has indicated that it forms a higher order structure upon either increase of the strand concentration or addition of K+. Gel electrophoresis showed that a high molecular weight species is formed in the presence of K+. The NMR study revealed a nearly 1:1 equilibrium of two kinds of higher order structures at the strand concentration of 2.1 mM and 100 mM Na+. This equilibrium shifted drastically upon addition of K+, and only one structure remained at 40 mM K+. It is suggested that the structure formed by r(GGAGGAA) in the presence of K+ is a new type of RNA quadruplex, where an A-quartet stabilized by mutual hydrogen bonding among the four adenine bases is sandwiched by conventional G-quartets.

Adenine↗

Dopamine depletion in nigrostriatal neurons in the genetically diabetic rat.

Employing tyrosine hydroxylase (TH) immunohistochemistry, we have revealed that the enzyme is depleted in nigrostriatal neurons in the genetically diabetic BB Wistar (BBWd) rat. A similar, but much less marked reduction in TH immunoreactivity occurred in the mesolimbic dopamine system, which arises from the ventral tegmental area and terminates in the nucleus accumbens. The projection to the olfactory tubercle appeared unaffected. Since the decreased levels of dopamine in the mesencephalic dopamine systems could not be restored under insulin-controlled conditions, the impairment of dopamine biosynthesis in the BBWd rat may be genetically programmed.

Animals↗

A role for CD4+ T cells in the pathogenesis of skin fibrosis in tight skin mice.

The tight skin (Tsk/+) mouse represents a murine model of heritable fibrosis with some similarities to the skin fibrosis seen in human scleroderma. Tsk/+ animals display alterations in connective tissue in some internal organs. Skin fibrosis can be adoptively transferred to normal recipients with Tsk/+ bone marrow or spleen cells and older Tsk/+ animals develop autoantibodies against topoisomerase suggesting that some of the pathogenesis in the Tsk/+ mouse may be mediated by autoimmunity. To determine the role of T cell subsets in the pathogenesis of fibrotic disease, Tsk/+ mice were bred with CD4- and CD8-deficient (CD4-/- and CD8-/-) mice. Tsk/+ CD4-/- mice showed a marked reduction in skin fibrosis as well as decreased cellularity and only mild collagen disorganization as compared to Tsk/+ CD4+ CD8+ control mice yet did not differ from Tsk controls in the level of serum anti-topoisomerase activity. In contrast, Tsk/+ CD8-/- mice exhibited the same histology in the skin as Tsk/+ controls yet had significantly reduced levels of serum anti-topoisomerase activity. Lung pathology, i.e. emphysema, was unaffected by both the CD4 or CD8 mutations. These data show that only some of the pathological effects of the Tsk mutation are T cell dependent and that different T cell subsets affect different parameters in this multi-organ model of fibrotic disease.

Animals↗

Divergent effects of intravenous dobutamine and nitroprusside on left atrial contribution to ventricular filling in dogs with chronic heart failure.

The left atrial (LA) contribution to left ventricular (LV) filling is often attenuated in patients with heart failure. It remains uncertain, however, whether therapy with positive inotropic agents or vasodilators improves or further impairs this maladaptation. In the present study, the effects of intravenous dobutamine and nitroprusside on the LA contribution to LV filling was examined in seven dogs with chronic heart failure produced by multiple sequential intracoronary microembolizations. Pulsed Doppler echocardiography was used to measure mitral inflow velocity before and after an intravenous infusion of dobutamine (4 micrograms/kg/min) and an intravenous infusion of nitroprusside (3 micrograms/kg/min). The percent LA contribution to LV filling was calculated as the ratio of the time-velocity integral of the LA component of mitral inflow velocity (Ai) to the time-velocity integral of total diastolic inflow velocity (Ti) times 100. Dobutamine increased LV filling pressure, LV end-diastolic wall stress, LV end-diastolic stiffness, and Ei, but had no effect on Ai or the percent LA contribution to filling (14% +/- 3% vs 12% +/- 2%) (p < 0.34). In contrast, nitroprusside decreased LV filling pressure, LV end-diastolic wall stress, and end-diastolic stiffness, and increased Ei, Ai, and the percent LA contribution to LV filling (12% +/- 2% vs 17% +/- 2%) (p < 0.01). The results indicate that dobutamine and nitroprusside have divergent effects on the LA contribution to LV filling. In dogs with chronic heart failure, dobutamine appears to impair LA contribution to the LV filling by augmenting LA workload, whereas nitroprusside appears to elicit greater LA contribution to LV filling by reducing the LA workload.

Animals↗

Effect of ooplast activation on the development of oocytes following nucleus transfer in cattle.

We assessed the effect of ooplast (enucleated oocytes) activation prior to receiving a donor nucleus on the development of nucleus transferred oocytes in cattle. The ooplasts were activated by electric stimulus at 30, 33, 36 and 39 h after being placed in culture medium for meiotic maturation. The activated ooplasts were further cultured in vitro, for a total 42 h from the beginning of maturation, 16- to 32-cell stage embryos produced by in vitro fertilization were used as donor embryos. The nucleus transferred oocytes were co-cultured with bovine oviductal epithelial cells in vitro. The fusion rate was not different between the activated (90%) and aged (94%) ooplasts 42 h after culture. Activated ooplasts receiving a donor nucleus showed a higher developmental rate than the aged ooplasts. Maximal development of the oocytes was obtained if the ooplast was activated at 9 h prior to receiving a donor nucleus. Thirty-nine percent developed to morulae and 24% to blastocysts. This compares (P<0.01) with 13% of the aged ooplasts developing to morulae and 8% to blastocysts. Of the activated ooplasts at 3, 6 and 12 h prior to fusion with a donor blastomere, 12, 16 and 13% developed to blastocysts, respectively. Of the 17 recipient cows receiving nucleus transferred embryos, 9 (53%) were diagnosed pregnant by palpation per rectum examination, and 3 normal offspring were obtained.

Journal Article↗

Left ventricular wall motion abnormalities in patients with subarachnoid hemorrhage: neurogenic stunned myocardium.

OBJECTIVES: The purpose of this study was to determine whether a relation exists between electrocardiographic (ECG) abnormalities and left ventricular wall motion in patients with subarachnoid hemorrhage. BACKGROUND: Although ECG changes simulating acute myocardial infarction are frequently seen in patients with subarachnoid hemorrhage, their relation to left ventricular wall motion has not been established. METHODS: Twelve patients with subarachnoid hemorrhage were classified according to the presence of ST segment elevation in at least two consecutive leads on admission: seven patients with ST segment elevation (group I) and five patients without ST segment elevation (group II). No patients had a previous history of heart disease. Left ventricular regional wall motion was evaluated by the centerline method. The mean (+/- SEM) duration from onset of subarachnoid hemorrhage to left ventriculography was 9 +/- 3 h in group I and 10 +/- 1 h in group II. Coronary angiography was performed to rule out wall motion abnormalities due to coronary artery disease while the ST segment was still elevated. Two-dimensional echocardiography was used to evaluate wall motion thereafter. RESULTS: All patients in group I showed ST segment elevation in ECG leads V4 to V6. Wall motion of the left ventricular apex was significantly reduced in group I compared with group II (-2.48 +/- 0.41 vs. -0.45 +/- 0.72, p < 0.02). No patients showed organic stenosis or vasospasm, or both, of epicardial coronary arteries. Wall motion abnormalities decreased echocardiographically in all patients, but one patient in group I died in hospital at 2 or 3 weeks after the onset of subarachnoid hemorrhage, when the T wave was inverted in leads V4 to V6. CONCLUSIONS: These findings suggest that patients with subarachnoid hemorrhage and ST segment elevation may demonstrate transient corresponding regional wall motion abnormalities. The mechanism of neurogenic stunned myocardium was not clearly elucidated in the present study.

Aged↗

Effects of low-power laser irradiation on multiple unit discharges induced by noxious stimuli in the anesthetized rabbit.

This study discusses the effects of low-power laser irradiation upon multiple unit discharges within a peripheral nerve that were induced by a noxious stimulation in anesthetized rabbits. Responding to a pinch stimulation of plantar skin, transitory increase of neuronal discharges was induced in the sural nerve. This was reduced within a minute, and persistent increases, which continued during a period of the stimulation, occurred. These increases became significantly smaller than the control value during a low-power laser irradiation that was applied to the exposed sural nerve distal to the recording site. These results suggested an inhibitory effect of low-power laser irradiation on the impulse conduction within a peripheral nerve. Possible analgesic effects of low-power irradiation are discussed.

Animals↗

Immunohistochemical localization of lysyl oxidase in normal human skin.

Lysyl oxidase (EC 1.4.3.13), a copper-dependent enzyme which catalyses the formation of aldehyde cross-links, and acts primarily on collagen and elastin, is known to be increased during wound healing and in fibrotic disorders including liver cirrhosis and atherosclerosis, and to be decreased in some hereditary connective tissue diseases and in malignant cell lines. A recent study showed that lysyl oxidase might possess tumour suppressor activity as an antioncogene for ras. Little is known about the localization of this enzyme in human skin. In this study, we determined immunohistochemically the localization of lysyl oxidase in normal skin of young and elderly subjects obtained from sun-exposed and unexposed regions of the body. All skin samples tested had similar distributions of lysyl oxidase. The enzyme was present both extracellularly and intracellularly. Extracellularly, a few granular aggregates of immunoreactants were observed along collagen and elastic fibres. These granules were more common in the adventitial portion of the dermis than in the reticular portion. Of all sun-exposed and unexposed regions studied, the skin of the face displayed the greatest amount of extracellular immunoreactants. Immunopositive granules were observed intracellularly in fibroblasts, vascular endothelial cells, sweat glands, sebaceous glands, arrector pili muscles and some keratinocytes. These findings provide evidence that, as suggested in recent reports, lysyl oxidase may have a variety of intracellular functions.

Adolescent↗

Lymphocyte function-associated antigen-1 is required for maximum elicitation of allergic contact dermatitis.

Cell adhesion molecules (CAMs) are known to play an important role in cell-mediated immunity and inflammation. To investigate the importance of one of the CAMs, lymphocyte function-associated antigen-1 (LFA-1) in allergic contact dermatitis (ACD), a specific anti-LFA-1 monoclonal antibody was injected into the ears of mice after sensitization but prior to challenge with dinitrofluorobenzene (DNFB). BALB/c mice were sensitized on the dorsal skin with DNFB, and challenged on the ears 6 days later. The effect of the antibody on the elicitation phase of contact sensitization was determined by its intradermal injection into the pinnae of the mice at doses of 1-40 micrograms. At 24 and 48 h after challenge, ear swelling was dose-dependently suppressed by injection of LFA-1 antibody at doses of 2-40 micrograms, which are less than 10% of the systemic dose required for effective suppression of delayed-type hypersensitivity in previous studies. Maximal inhibition of ear swelling (44%) was observed after injection of 20 micrograms of anti-LFA-1 antibody. Inhibition of ear swelling was accompanied by a reduction in dermal oedema and leucocyte infiltration into the dermis. Our results suggest that LFA-1 plays a significant role in the elicitation of ACD reactions, and also that our test system is a sensitive and useful means of evaluating the blocking effect of antibody to adhesion molecules thought to be involved in the elicitation phase of ACD.

Animals↗

Differential modulation of interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) in human epidermal keratinocytes by UVB.

Conflicting reports exist concerning ultraviolet-B (UVB) effects on keratinocyte (KC) interleukin-1 (IL-1) expression. To clarify the modulatory effects of UVB on IL-1, the following study was undertaken. Normal human epidermal KCs cultured in a standard low Ca2+ and serum-free medium were irradiated in quiescent phase with UVB. In this study, we used semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to determine the mRNA level of interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta). After exposure to 100 or 300 J/m2 UVB, a transient increase in mRNA levels was observed within 1 hour for IL-1 alpha and 3 to 6 h for IL-1 beta. Following this transient induction, mRNA levels for both IL-1 alpha and IL-1 beta returned to steady-state levels after 100 J/m2. After 300 J/m2 irradiation, IL-1 alpha and IL-1 beta levels were downregulated compared to unirradiated cultures at 24-h post-irradiation. The half-life for IL-1 alpha and IL-1 beta was estimated using actinomycin D treatment. Both IL-1 alpha and IL-1 beta mRNAs half-lives (t1/2) decreased faster in irradiated cells (t1/2 = 30 minutes for IL-1 alpha and 2 h for IL-1 beta) compared to unirradiated cells (t1/2 = 1 h and 4 h, respectively). These results suggest that IL-1 alpha and IL-1 beta mRNA expression are differentially regulated by UVB. In contrast to down-regulation of mRNA levels, a significant increase in IL-1 alpha protein levels, measured by ELISA, was observed in culture supernatants from 6 h to 24 h after 300 J/m2 UVB irradiation. Cycloheximide treatment did not abrogate this increase in IL-1 alpha protein level. Since this dose of UVB irradiation decreased the stability of IL-1 alpha and IL-1 beta mRNA, this suggests that the release of IL-1 alpha after UVB irradiation was due to leakage from UVB-damaged cells and not from de novo protein synthesis.

Cells, Cultured↗

Effects of long-term monotherapy with enalapril, metoprolol, and digoxin on the progression of left ventricular dysfunction and dilation in dogs with reduced ejection fraction.

BACKGROUND: Recent clinical trials have suggested that therapy with angiotensin-converting enzyme inhibitors in asymptomatic patients with reduced left ventricular (LV) function can significantly reduce the incidence of congestive heart failure compared with patients receiving placebo. In the present study, we examined the effects of long-term monotherapy with enalapril, metoprolol, and digoxin on the progression of LV systolic dysfunction and LV chamber enlargement in dogs with reduced LV ejection fraction (EF). METHODS AND RESULTS: LV dysfunction was produced in 28 dogs by multiple sequential intracoronary microembolizations. Embolizations were discontinued when LVEF was 30% to 40%. Three weeks after the last embolization, dogs were randomized to 3 months of oral therapy with enalapril (10 mg twice daily, n = 7), metoprolol (25 mg twice daily, n = 7), digoxin (0.25 mg once daily, n = 7), or no treatment (control, n = 7). As expected, in untreated dogs, LVEF decreased (36 +/- 1% versus 26 +/- 1%, P < .001) and LV end-systolic volume (ESV) and end-diastolic volume (EDV) increased during the 3-month follow-up period (39 +/- 4 versus 57 +/- 6 mL, P < .001, and 61 +/- 6 versus 78 +/- 8 mL, P < .002, respectively). In dogs treated with enalapril or metoprolol, LVEF remained unchanged or increased after therapy compared with before therapy (35 +/- 1% versus 38 +/- 3% and 35 +/- 1% versus 40 +/- 3%, respectively, P < .05), whereas ESV and EDV remained essentially unchanged. In dogs treated with digoxin, EF remained unchanged but ESV and EDV increased significantly. CONCLUSIONS: In dogs with reduced LVEF, long-term therapy with enalapril or metoprolol prevents the progression of LV systolic dysfunction and LV chamber dilation. Therapy with digoxin maintains LV systolic function but does not prevent progressive LV enlargement.

Animals↗

[Clinical evaluation of intracavernous self-injection of vasoactive drugs for impotence: a long-term follow-up observation].

From December 1989 to September 1992, nine patients with impotence were instructed to perform intracavernous self-injection of vasoactive drugs. At first 40 mg of papaverine hydrochloride was used in all patients and the response on erection was evaluated. If the response did not show sufficiently functional erection, a mixture of 40 mg of papaverine hydrochloride and 1 mg of phentolamine mesylate or 20 mg of prostaglandin E1 was reinjected. Eight patients had achieved full erections and vaginal penetrations without noteworthy complications during the follow-up period. Out of eight patients, three patients were able to ejaculate and one patient showed recovery of erection. No major side effects were seen. In conclusion, intracavernous self-injection is a useful modality for impotence.

Adult↗

[A case of resected gastric cancer in which cancer had apparently disappeared by preoperative administration of tegafur].

This paper reports a case of gastric carcinoma in which cancerous tissue was not found in the resected specimen, apparently due to preoperative tegafur administration. The patient was a 56-year-old woman who was diagnosed by initial endoscopic examination including endoscopic biopsy as having an advanced gastric cancer of Borrmann type 2. During the waiting period for operation, she was placed on oral administration of tegafur (Futrafur capsule) 600 mg 3 times a day for 31 days. The second endoscopic examination 14 days later showed reduction of both marginal elevation and central excavation of the lesion, and biopsy at this time was negative for carcinoma. Because of the initial definite diagnosis of cancer, a gastric resection was performed 31 days later, but no cancerous tissue was found in the resected specimen. This case was considered to be a carcinoma especially sensitive to tegafur.

Adenocarcinoma↗

Effects of a novel topical immunomodulator, imiquimod, on keratinocyte cytokine gene expression.

A novel immunomodulator, imiquimod, has been shown to be an effective topical antiviral and antitumor agent in animal models. Imiquimod has been reported to induce interferon-alpha and other cytokines in animals and humans, but its precise role as an immunomodulator at skin sites has not been determined. We investigated its effect on cytokine gene expression in the human epidermal carcinoma cell line COLO-16 and human keratinocytes. COLO-16 cells were incubated with imiquimod (1 and 10 micrograms/ml) and human keratinocytes with 5 micrograms/ml for 6 or 24 h. Cytokine gene expression was analyzed by reverse-transcriptase PCR. In COLO-16 cells, imiquimod stimulated IL-6 mRNA levels 2.3- and 4.4-fold at 1 and 10 micrograms/ml after 6 h. IL-8 mRNA increased 4-fold at both 1 and 10 micrograms/ml. At 24 h, though IL-6 mRNA level at 1 micrograms/ml was further stimulated, enhanced expressions of IL-8 at 1 micrograms/ml and both IL-6 and IL-8 at 10 micrograms/ml were down-regulated. In human keratinocytes, 5 micrograms/ml of imiquimod stimulated IL-6 mRNA levels 1.4-fold at 6 h and 2.1-fold at 24 h, and IL-8 mRNA levels 1.7- and 2.0-fold at 6 and 24 h. IL-1 alpha mRNA levels in COLO-16 or keratinocytes were unchanged by either dose or incubation time. These results suggest that stimulation of IL-6 and IL-8 expression may be involved in the immunomodulating action of imiquimod.

Adjuvants, Immunologic↗