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Biomedical subjects

H Tsuchiya

Publications and source records attributed to H Tsuchiya.

At least 55 records · Page 3Linked to original sources

Reduction of membrane fluidity by antibacterial sophoraflavanone G isolated from Sophora exigua.

Sophoraflavanone G (5,7,2',4'-tetrahydroxy-8-lavandulylflavanone) has been referred as a phytochemical with the intensive antibacterial activity. To elucidate the pharmacological mechanism underlying an antibacterial action, sophoraflavanone G was isolated from Sophora exigua, thereafter its effect on membrane fluidity was studied using model membranes and compared with less active naringenin lacking 8-lavandulyl and 2'-hydroxyl groups. Highly purified sophoraflavanone G of 0.05-5 microg/ml, corresponding to the minimum growth inhibitory concentrations against various bacteria, significantly increased fluorescence polarization of the liposomes prepared from 1,2-dipalmitoyl-L-alpha-phosphatidylcholine and 1-palmitoyl-2-oleoyl-L-alpha-phosphatidylcholine. Such increases were found in both liposomes measured with two fluorescent probes to indicate an alteration of membrane fluidity in hydrophilic and hydrophobic regions, suggesting that sophoraflavanone G reduces the fluidity of outer and inner layers of membranes. Although naringenin also showed the membrane effect, it needed concentrations over 2.5 microg/ml to induce a significant reduction of membrane fluidity. Sophoraflavanone G is considered to exert an antibacterial effect by reducing the fluidity of cellular membranes.

1,2-Dipalmitoylphosphatidylcholine↗

Event-related brain potentials in response to novel sounds in dementia.

OBJECTIVE: Non-target, deviant stimuli generate an earlier latency, front-central novelty P3, whereas correctly detected task-relevant stimuli generate a parietal maximal target P3. We examined whether the P3 component to novel stimuli is affected by dementing processes, and is therefore useful for distinguishing Alzheimer's type dementia (AD) from vascular dementia (VD). METHODS: We recorded ERPs to task-relevant stimuli (target P3) and novel task-irrelevant stimuli (novelty P3) in an auditory oddball task in AD (n = 16), VD (n = 16), and age-matched controls (n = 18). The amplitude, latency, and scalp topography of target and novelty P3 were compared among 3 groups using ANOVA. The relationship between P3 measures and intelligence scores were evaluated by correlation analysis. RESULTS: The amplitude, latency and scalp topography of the target P3 were comparably affected by both AD and VD. However, the amplitude of the novelty P3 was markedly reduced in VD, but not in AD, and the scalp topographics were different in the 3 groups. The amplitude was maximal at frontal sites in controls, at central sites in AD, and at parietal sites in VD. The target P3 latency was prolonged in both AD and VD, whereas the novelty P3 latency was only prolonged in VD. AD was discriminated satisfactorily from VD by using the novelty amplitude at Cz and the ratio of the amplitudes at Fz and Pz as independent variables. CONCLUSIONS: These results suggest that the response to novel stimuli is differentially affected by dementia with degenerative and vascular etiology.

Acoustic Stimulation↗

A possible link between beta -carboline metabolism and infantile autism.

Benzodiazepine receptors and abnormal hepatic metabolism have been suggested to participate in several neuropsychiatric disorders including autism. Neuropsychoactive beta-carboline alkaloids as the potent ligands for benzodiazepine receptors are endogenously produced and exogenously supplied much more than benzodiazepines. 1-Methyl-1,2,3,4-tetrahydro-beta-carboline, a predominant alkaloid in humans and foodstuffs, is metabolically hydroxylated in liver. Although its in vivo levels show no difference between autistic and healthy children, the metabolic 6-hydroxylation is significantly decreased in autistic subjects. Therefore, it could be hypothesized that the reduced hepatic metabolism of 1-methyl-1,2,3, 4-tetrahydro-beta-carboline to 6-hydroxyl metabolite may be linked to the pathogenesis of infantile autism as suggested for autistic occurrence to involve the pathology similar to hepatic encephalopathy.

Autistic Disorder↗

Malignant transformation of a giant cell tumor 25 years after initial treatment.

The current authors report a patient in whom a malignant fibrous histiocytoma developed long after a benign giant cell tumor of bone was removed from the same site. Twenty-five years after a benign giant cell tumor of the lateral condyle of the proximal tibia had been treated by curettage and iliac bone grafting without radiotherapy, a 53-year-old woman noted progressive pain and an enlarging mass in the same area. Radiographs showed osteolytic change, whereas magnetic resonance imaging indicated a tumor arising at the site of the giant cell tumor and extending beyond the bone. Examination of an open biopsy specimen showed a high-grade malignant fibrous histiocytoma with some areas rich in giant cells. After five courses of caffeine assisted intraarterial chemotherapy, the tumor was resected with an adequate margin, and the defect was reconstructed with an implanted prosthesis of corresponding shape. The extensor mechanism of the knee was reinforced using an allograft of fascia from the tensor fascia lata muscle. The resected specimen showed a good histologic response (95% tumor necrosis) to preoperative chemotherapy. Excellent function in the knee has been regained with no evidence of disease recurrence. Caffeine potentiated chemotherapy was effective in minimizing the extent of tumor excision, in this case of high-grade malignant fibrous histiocytoma representing transformation from a benign giant cell tumor.

Adult↗

Effect of estrogen and progesterone on gene expression of growth regulatory molecules and proto-oncogene in vascular smooth muscle cells.

We examined influences of estrogen and progestogen on gene expression of the growth regulatory molecules: platelet-derived growth factor (PDGF), interleukin-1 (IL-1), interleukin-6 (IL-6) and proto-oncogene c-myc in vascular smooth muscle cells (VSMC) by reverse transcription-polymerase chain reaction (RT-PCR) and Southern-blotting. VSMC were exposed to estrone-sulfate (E1-S) and medroxyprogesterone acetate (MPA) to induce differentiation. E1-S inhibited the expression of PDGF-A chain, IL-1, IL-6 and c-myc mRNA, whereas MPA had no effect. Inhibition by E1-S was not affected by treatment combined with MPA. These findings suggest that estrogen modulates these growth regulatory molecules and c-myc gene expression in VSMC but not progestogen. We concluded that estrogen may have a direct atheroprotective effect through inhibition of growth regulatory factors.

Arteriosclerosis↗

Troglitazone suppresses cell growth of myeloid leukemia cell lines by induction of p21WAF1/CIP1 cyclin-dependent kinase inhibitor.

In a human eosinophilic leukemia cell line, EoL-1, cell proliferation was suppressed by 2-day treatment with troglitazone. EoL-1 cells treated with troglitazone were arrested and maintained in the G0/G1 phase in the cell cycle. This suppression correlated with the up-regulation of mRNA for p21WAF1/CIP1 cyclin-dependent kinase (Cdk) inhibitor. The inhibitory effects of troglitazone on cell proliferation and expression of p21 mRNA were observed in a human myelomonocytic cell line, U937, and a human myelomonoblastic cell line, KPB-M15. In addition, in EoL-1 cells, p21 protein was induced by troglitazone treatment and the induction was inhibited by protein synthesis inhibitor, cycloheximide. These data suggest that troglitazone inhibits cell proliferation in myeloid leukemia cell lines at least in part by induction of p21 Cdk inhibitor.

Antineoplastic Agents↗

A heterozygous mutation of beta-actin associated with neutrophil dysfunction and recurrent infection.

A human disorder caused by mutation in nonmuscle actin has not been reported. We report here a variant of nonmuscle actin in a female patient with recurrent infections, photosensitivity, and mental retardation. She also had abnormalities in neutrophil chemotaxis, superoxide production, and membrane potential response. Two-dimensional PAGE analysis of proteins from neutrophils and other cell types from this patient demonstrated a unique protein spot migrating at 42 kDa with pI shifted slightly to neutral relative to normal beta- and gamma-actin. Digestion peptide mapping and Western blotting showed this spot to be an abnormal actin. A full-length cDNA library was constructed by using mRNA from patient's cells and cDNA encoding the mutant beta-actin molecule was identified by an in vitro translation method. Sequencing of the clones demonstrated a G-1174 to A substitution, predicting a glutamic acid-364 to lysine substitution in beta-actin and eliminating a HinfI DNase restriction site found in normal beta-actin sequence. By HinfI digestion and by sequencing, the mutation in one allele of patient's genomic DNA was confirmed. Though no defect in cell-free polymerization of actin was detected, this defect lies in a domain important for binding to profilin and other actin-regulatory molecules. In fact, the mutant actin bound to profilin less efficiently than normal actin did. Heterozygous expression of mutant beta-actin in neutrophils and other cells of this patient may act in a dominant-negative fashion to adversely affect cellular activities dependent on the function of nonmuscle actin.

Actins↗

Transitional-cell carcinoma of the bladder in an adolescent.

We report a rare case of transitional-cell carcinoma of the bladder in an 18-year-old girl with intermittent hematuria. The tumor was diagnosed through the cystogram phase of the excretory urogram and bladder ultrasound. Transurethral resection (TUR) of the tumor was performed. As most tumors are low-grade and low-stage and are associated with an excellent prognosis, endoscopic TUR is the preferred therapeutic measure. However, recurrence as well as death have been reported in this age group. These patients should be followed carefully, similar to adult patients.

Adolescent↗

Antiplatelet activity of soy sauce as functional seasoning.

In seeking the functionality of foodstuffs applicable to medicine, soy sauce was found to show antiplatelet activity. Therefore, the active components in soy sauce were purified, structurally identified, and studied for their inhibitory effects on the aggregation of human platelets. Aqueous 2-fold diluents of soy sauce inhibited platelet aggregation induced by collagen and epinephrine depending on the dilution factor. Since a basic extract with diethyl ether completely inhibited collagen-induced aggregation, it was subjected to serial extractions and multistep HPLC fractionations for purifying antiplatelet components. The finally obtained isolates were identified as 1-methyl-1,2,3,4-tetrahydro-beta-carboline and 1-methyl-beta-carboline on the basis of EI-MS, (1)H NMR, diode array, and fluorescence spectra. Their spectral data and chromatographic behaviors were the same as those of synthetic ones. 1-Methyl-1,2,3, 4-tetrahydro-beta-carboline showed mean concentrations (n = 5-6) of 4.6, 4.2, 28.6, 11.6, and 65.8 microgram/mL to produce 50% inhibition of the maximal aggregation response induced by epinephrine, platelet-activating factor, collagen, adenosine 5'-diphosphate, and thrombin, respectively. Its inhibitory effect was much greater than that of 1-methyl-beta-carboline on platelet aggregation by all the tested inducers. The quantitative HPLC analysis revealed that the significant amounts of both antiplatelet compounds were uniformly contained in commercially available soy sauce. From these results, soy sauce may be referred to as functional seasoning containing alkaloidal components with the potent preventive effect on thrombus formation.

Carbolines↗

Rat macrophage inflammatory protein-1alpha, a CC chemokine, acts as a neutrophil chemoattractant in vitro and in vivo.

Recombinant rat macrophage inflammatory protein-1alpha (rMIP-1alpha) at a concentration of 3x10(-8) M had strong neutrophil chemotactic activity, though the potency of rMIP-1alpha was less than that of cytokine-induced neutrophil chemoattractant (CINC)-1 at lower concentrations. In addition, rMIP-1alpha induced neutrophil chemotaxis in vivo when rMIP-1alpha was injected into the preformed air-pouch on the back of rats. The adhesion of rMIP-1alpha-treated neutrophils to fibrinogen significantly increased, reaching a maximum adhesion at 10(-8) M. Stimulation of neutrophils with rMIP-1alpha induced a transient increase in intracellular free [Ca2+] dose-dependently. rMIP-1alpha still induced an increase in the intracellular [Ca2+] of rat neutrophils stimulated first with CINC-1, CINC-3 or C5a, suggesting that rat neutrophils have a specific receptor for rMIP-1alpha. Supporting these findings, an additive increase in chemotactic potency was found when both rMIP-1alpha and CINC-were added to the lower wells of Boyden chamber in vitro. In addition, high levels of rMIP-1alpha were detected in the inflammatory site of air-pouch/carrageenan-induced inflammation in rats. Our results suggest that rMIP-1alpha acts as a neutrophil chemoattractant and, together with CINCs, plays an important role in infiltration of neutrophils into inflammatory sites in rats.

Animals↗

High-performance liquid chromatographic analysis of chlorhexidine in saliva after mouthrinsing.

A high-performance liquid chromatographic method was developed to quantify chlorhexidine in human saliva. After addition of an internal standard and acidic-deproteinization, saliva samples were chromatographed by the reversed-phase ion-pair system using pentanesulfonate as a counterion and the eluates were detected by a diode array detector. Under optimal conditions, the baseline separation of analytes was achieved within 4.5 min without interference from components in saliva matrix. The method showed high selectivity to salivary chlorhexidine, quantitative range (50. 0ng/ml-50.0 microg/ml), recovery (96.0-97.8%) and analytical precision (intra- and interassay CVs within 0.41%), permitting the effective application to both saliva and aqueous solutions. Chlorhexidine was found in saliva at microgram per milliliter levels for at least 8 h after mouthrinsing with 10 ml of an aqueous solution of chlorhexidine (0.1 or 1.0 mg/ml) for 1 min. The concentrations of salivary chlorhexidine were reduced by ingesting the acidic beverage and food, but not by ingesting the neutral beverage. The adsorption experiments in artificial saliva revealed that chlorhexidine was significantly adsorbed to saliva-coated hydroxyapatite, buccal epithelial cells and mucin. The proposed method will be a useful tool to study salivary chlorhexidine after oral application and its retention in the oral cavity.

Adsorption↗

Effects of green tea catechins on membrane fluidity.

Catechins originating from green tea have been used in plaque inhibition for caries prevention and treatment for liver damage because of their antibacterial activity against cariogenic bacteria and protective activity on hepatic cells. The effects of catechins on membrane fluidity were studied by a fluorescence polarization method using liposomes prepared with dipalmitoylphosphatidylcholine and dioleoylphosphatidylcholine to assess their pharmacological mechanism at micromol/l levels found in human body fluids after clinical application. All eight catechins tested, ranging from 1 to 1,000 micromol/l, significantly reduced membrane fluidity in both hydrophilic and hydrophobic regions of lipid bilayers. Catechin gallate esters were superior in fluidity reduction to the corresponding nonesters. The fluidity-reducing degree was different between the cis and trans forms, suggesting the stereospecific activity of catechins. A reference antiplaque agent, chlorhexidine, similarly reduced membrane fluidity at the antibacterial concentration. (+)-Catechin (250 micromol/l) and (-)-epigallocatechin gallate (2.5 micromol/l) significantly prevented the membrane fluidization induced by hepatotoxic chloroform. These results indicate that the reduction in membrane fluidity is responsible for the antiplaque and hepatoprotective effects of green tea catechins.

1,2-Dipalmitoylphosphatidylcholine↗

Transplantation of osteoblast-like cells to the distracted callus in rabbits.

We carried out limb lengthening in rabbits and then transplanted osteoblast-like cells derived from the tibial periosteum to the centres of distracted callus immediately after distraction had been terminated. Two weeks later the transaxial area ratio at the centre of the distracted callus and the bone mineral density (BMD) were significantly higher in the transplanted group, by 21% and 42%, respectively, than in the non-injected group or the group injected with physiological saline (p < 0.05). Callus BMD as a percentage of density in uninvolved bone was also significantly higher in the transplanted group (p < 0.05) than in the other two groups, by 27% and 20% in the second and fourth weeks, respectively (p < 0.05). Mechanically, the callus in the transplanted group tended to be stronger as shown by the three-point bending test although the difference in fracture strength was not statistically significant. Our results show that transplantation of osteoblast-like cells promotes maturity of the distracted callus as observed at the second and fourth weeks after lengthening. The method appears promising as a means of shortening the consolidation period of callus distraction and decreasing complications during limb lengthening with an external fixator.

Absorptiometry, Photon↗

[Clinical effect of arotinolol hydrochloride and its influence on renal function in elderly patients with essential hypertension].

Arotinolol hydrochloride with alpha-and beta-receptor blocking action, developed in Japan, is mainly used for the treatment of hypertension. The study population consisted of 42 outpatients with essential hypertension with a blood pressure greater or equal to 160/96 mmHg. 10 men and 32 women, with a mean age of 77.5 year. The patients received 10 mg arotinolol hydrochloride daily for 24 weeks which was taken orally twice a day. We evaluated the changes of blood pressure, heart rate and chief complaints of patients before and every 4 weeks during treatment and the renal function before, 12 weeks after and 24 weeks after, the administration of arotinolol hydrochloride. Blood pressure and heart rate decreased significantly after 4 weeks of treatment with arotinolol hydrochloride (p < 0.05). However, no significant changes were found in blood urea nitrogen, serum creatinine, serum albumin, beta2-microglobuline, NAG or creatinine clearance during the 24 weeks of treatment. These results indicate that arotinolol hydrochloride has antihypertensive effects without renal dysfunction in elderly patients with essential hypertension.

Adrenergic alpha-Antagonists↗

Impaired endothelial function in hypertensive elderly patients evaluated by high resolution ultrasonography.

BACKGROUND: Multiple investigations both in experimental models and in middle-aged patients with essential hypertension have demonstrated impaired endothelium-dependent vasodilation. OBJECTIVE: To determine whether hypertension exerts an additional negative effect on endothelial function of large arteries in hypertensive elderly patients who may already be affected by endothelial dysfunction due to aging. PATIENTS AND METHODS: Thirteen elderly patients with hypertension (69 9 years of age [mean SD]) were compared with 13 matched healthy elderly subjects (72 6 years of age). High resolution vascular ultrasound was used to measure brachial artery responses to reactive hyperemia (with increased flow causing endothelium-dependent dilation) and to sublingual nitroglycerine (causing endothelium-independent dilation). RESULTS: Flow-mediated diameter (FMD) was significantly impaired in the hypertensive elderly group (6.7 3.3% versus 13.3 3.8% in the control group, P<0.05). No significant difference could be found in nitroglycerine-induced dilation between the elderly control group (12.1 4.9%) and the hypertensive elderly (10.2 6.8%). On simple linear analysis, FMD was inversely correlated with age (r=-0.60, P=0. 03) in the healthy elderly group. FMD in the hypertensive elderly was inversely related to age (r=-0.41, P=0.04) and mean blood pressure (r=-0.67, P=0.01). CONCLUSIONS: This study showed decreased FMD with aging even in the healthy elderly, with a further decline in hypertensive elderly compared with healthy elderly subjects. This impairment of FMD in the hypertensive elderly group was related to age and mean blood pressure, indicating that aging and hypertension may impair endothelial function in the brachial artery of elderly patients with hypertension.

Age Factors↗

Marginal excision for osteosarcoma with caffeine assisted chemotherapy.

The authors report on intentional marginal excision for osteosarcoma in conjunction with caffeine assisted chemotherapy for the purpose of preservation of good limb function. Twenty-seven patients with osteosarcoma (22 patients with Stage IIB and five with Stage IIIB) preoperatively were given three-to-five courses of intraarterial cisplatin and caffeine without or with doxorubicin. For 26 (96%) responders to the chemotherapy, limb salvage surgery was conducted by means of an intentional marginal procedure, which led to the preservation of important structures such as major neurovascular bundles, tendons, ligaments, muscles, and the epiphysis. Tumors were located in the distal femur in 11 patients, the proximal tibia in eight, the proximal fibula in four, the proximal humerus in two, and the proximal femur in one patient. The histologic response of these 26 patients to the preoperative chemotherapy showed no viable cells in 19 patients with Stage IIB osteosarcoma and only scattered foci of viable cells in two patients with Stage IIB and five patients with Stage IIIB osteosarcoma. As for reconstruction, distraction osteogenesis was performed in eight patients, allograft or autoclaved bone and prosthesis composite in four, autoclaved bone in two, osteochondral allograft in two, megaprosthesis in six, and resection alone in four patients. The average functional evaluation of the 26 patients was 91% of normal. Local tumor recurrence was seen in one patient, whereas 18 patients with Stage IIB osteosarcoma remain diseasefree with a mean followup of 61 months. Two patients with Stage IIB osteosarcoma and four patients with osteosarcoma Stage IIIB died of the disease. Intentional marginal excision for osteosarcoma in conjunction with caffeine assisted chemotherapy is advantageous because it results in the preservation of healthy important structures, with joint preservation possible in selected cases. This approach should help to improve the success rate of limb salvage surgery for osteosarcoma and to preserve the function of the affected limb.

Adolescent↗

High-performance liquid chromatographic analysis of polyhydroxyflavones using solid-phase borate-complex extraction.

A high-performance liquid chromatographic method using a solid-phase borate-complex extraction pretreatment was studied for the selective quantitation of polyhydroxyflavones in vegetables, red wine and human blood plasma. Homogenate, extract and intact samples were applied to phenylboric acid cartridges to retain polyhydroxyflavones on the solid-phase by forming the borate-complex, followed by elution of the retained analytes with an acidic solvent. Reversed-phase chromatography with diode array detection allowed the simultaneous separation of rutin, myricetin, fisetin, morin, quercetin and kaempferol without significant interference from other components, indicating high selectivity of the solid-phase borate-complex extraction. The absolute recoveries of quercetin, fisetin and rutin were superior to those of kaempferol, myricetin and morin, suggesting a difference in the complex formation efficiency between 1,2- and 1,3-diol structures. When using fisetin as an internal standard, polyhydroxyflavones were quantified in the concentration range 0.10-30.0 microg/ml. In replicate spiking experiments with standards, the mean relative recoveries ranged between 75.7 and 104.6%, and the intra-and inter-assay C.V.s ranged between 0.8 and 10.2% for onion, wine and plasma samples. The proposed method will be applicable to nutritional and pharmacokinetic experiments of polyhydroxyflavones.

Chromatography, High Pressure Liquid↗