Biomedical subjects

T Uchida

Publications and source records attributed to T Uchida.

At least 433 records · Page 24Linked to original sources

PubMed1995

Relation between micellar structure of model bile and activity of esterase.

In a model bile solution composed of lecithin (L)-bile salt (B), the solubilization of lipid and the accessibility of enzyme to the lipid were examined by observation of EPR spectra and measurement of enzyme activity. The lifetime of the spin probe in the micellar phase was estimated to be approx. 1 microsecond by means of line shape analysis. Both population and lifetime increased with temperature and the molar ratio of lecithin to bile salt (L/B). The EPR data indicated that simple micelle of bile salt, mixed disk micelle of bile salt-lecithin, and multi-lamellar mixed disk micelle can exist in a model bile solution, depending on the L/B molar ratio across a range from 0 to 1.5. The maximal power of the mixed disk micelle to solubilize cholesteryl ester in the model bile at a L/B molar ratio of 1:1 was confirmed by EPR measurement of cholesteryl 12-DOXYL-stearate. Observation of the enzyme activity on a mixture of model bile and substrate at 37 degrees C revealed selective accessibility of cholesterol esterase (bovine pancreas) to mixed disk micelle, of cholesterol oxidase (Streptomyces cinnamomeus) to both simple and mixed disk micelle, and of pancreatic lipase (porcine pancreas) to both simple micelle and an oil droplet of substrate. The temperature-dependent activity of cholesterol oxidase to cholesterol in mixed disk micelle can be explained in terms of mesomorphic phase transition of lecithin side chains followed with fluidity of liquid crystal phase. Regarding phospholipase C from Bacillus cereus, though the selective accessibility to the micelles was not observed at 37 degrees C, a decrease in activity for mixed disk micelle could be found at lower temperatures.

Bile↗

PubMed1995

cDNA cloning of a novel brain-specific protein p25.

A cDNA for a bovine brain-specific protein p25 which had been originally found as a major protein in a partially purified fraction of tau protein kinases was cloned. The deduced amino-acid sequence consists of 218 amino acids (M(r) 23,472) and has no significant homology with previously reported proteins. p25 is a basic protein and has a consensus sequence for ATP-binding in the C-terminal region.

Amino Acid Sequence↗

PubMed1995

Early detection of prostate cancer. Results of a prostate specific antigen-based detection program in Japan.

BACKGROUND: Prostate cancer increasingly is becoming a medical problem in Japan. To maximize the chance for cure, extensive effort is being made to detect prostate cancer while confined to the gland. METHODS: Determination was made of serum prostate specific antigen (PSA) in the early detection of prostate cancer in an Asian male population aged 55 years or older. Recommendation for biopsy was based solely on this parameter when its value exceeded 2.0 ng/ml in IMx immunoenzymetric assay. RESULTS: Of 1189 men, 150 (12.6%) had elevated values. The proportion of males with serum PSA greater than 4.0 ng/ml was only 3.4%. Of the 99 males who underwent ultrasound-guided biopsies, 16 cancers were detected, a cancer detection rate of 1.3%. Most of these (81.3%) were clinically localized cancers. Seven patients with cancer had serum PSA levels below 4.0 ng/ml including 5 less than 3.0 ng/ml. These tumors would have been overlooked if the conventional cutoff levels of 4.0 ng/ml had been applied. Radical prostatectomy was conducted on 13 patients. All had histologic features of clinically significant cancer and 69.2% were pathologically confined within the gland. CONCLUSIONS: Elderly Asian males appear to have lower serum PSA than their Western counterparts. The optimal cutoff of serum PSA for early detection should be examined further in Asian male populations. Differences in the incidence of prostate cancer between ethnic groups may have been overestimated in previous studies owing in part to unawareness by the physician and general public. Though the frequent use of more sophisticated diagnostic modalities will likely disclose greater numbers of prostate cancers in Japan, whether early detection reduces mortality by minimizing risk of death from cancer remains a point to be determined.

Aged↗

PubMed1995

Genomic instability of microsatellite repeats in prostate cancer: relationship to clinicopathological variables.

Sixty-six patients with prostatic adenocarcinoma were screened for somatic instability at 8 microsatellite marker loci on 5 chromosomes. Differences in unrelated microsatellites for tumor and normal DNA were detected in 13 (19.7%) patients. Only extraglandular spread (nodal involvement and distant metastasis) was found to show significant association with somatic instability after controlling for other clinicopathological variables (P < 0.05). Microsatellite instability may possibly occur during the early stages of neoplastic transformation in a subset of prostate cancer rather than as a late event. This may be related to a phenotype with growth advantage. The frequency of this mutator phenotype is much higher in the United States than Japan, reflecting racial differences in the molecular tumorigenesis of this malignancy.

Adenocarcinoma↗

PubMed1995

Purification, cloning, and expression of murine uridine phosphorylase.

Uridine phosphorylase was purified 10,300-fold from tumors of the murine colorectal adenocarcinoma cell line, Colon-26. Degenerate DNA probes were synthesized corresponding to partial amino acid sequences and used to screen a Colon-26 cDNA library. A cDNA clone of 1327 base pairs that contains a 5' untranslated region, a coding region of 933 base pairs, and a 3' nontranslated region with a polyadenylated tail was identified. The cDNA was confirmed to be uridine phosphorylase by 1) sequence comparison to uridine phosphorylase of Escherichia coli, 2) substrate specificity studies with recombinant protein expressed in COS-7 cells that demonstrated relatively high enzyme activity with uridine as substrate compared low levels when thymidine was used, and 3) inhibition of enzyme activity by the competitive inhibitor 2,2'-anhydro-5-ethyluridine. Northern blot analysis using the cDNA as a probe, demonstrated high levels of mRNA expression in Colon-26. Expression was low in NIH3T3 cells, but high in DMBA-3 and PH-1 cells, which are NIH3T3-derived cells that have been transformed with mutated murine Ha-ras and viral Ha-ras, respectively. Expression of uridine phosphorylase mRNA in these cell lines was further enhanced by treating the cells with the inflammatory cytokines, tumor necrosis factor-alpha, interleukin 1 alpha, and interferon gamma.

Adenocarcinoma↗

PubMed1995

Microsatellite instability in prostate cancer.

Assessment of the genetic instability of a microsatellite has indicated a new mechanism in human carcinogenesis. Examination was made to determine whether microsatellite instability is associated with the onset of prostate cancer. Twenty-nine DNA samples from 24 primary prostate cancer, two metastatic lymph-node and three benign prostatic hypertrophy patients were used. Differences in unrelated microsatellites for tumor and normal DNA were detected in nine of 24 (37.5%) cases. Seven of 11 (63.6%) with poorly differentiated adenocarcinomas and seven of 15 (46.7%) stage D metastatic patients showed somatic instability in a number of microsatellites. Statistically significant differences in well to moderately differentiated tumors and poorly differentiated cancer (P = 0.015, Chi-square test), were detected but not for stages A-C and D (P = 0.2311). Genetic alterations would thus appear to be rare in low grade and/or early prostate cancers but more common in high grade and/or advanced prostate cancers.

DNA Replication↗

PubMed1995

Porcine brain neurofilament-H tail domain kinase: its identification as cdk5/p26 complex and comparison with cdc2/cyclin B kinase.

Using dephosphorylated neurofilament (NF) proteins as substrates, the kinase with a higher activity for the dephosphorylated NF-H than the phosphorylated form of NF-H was searched for in the porcine brain extract. Most NF-H kinase activity in the brain extract pelleted with microtubules. The NF-H kinase purified from a high salt extract of the microtubule pellets was composed of cdk5 and a 26 kDa protein, a fragment of the 35 kDa regulatory subunit of cdk5. In contrast to the association of the active kinase with microtubules, each of uncomplexed cdk5 and the 35 kDa regulatory subunit was differently distributed in the supernatant fraction and the pellet, respectively, by ultracentrifugation of the brain extract. Dephosphorylated forms of NF-H and NF-M became reactive to antibodies recognizing in vivo phosphorylation sites (SMI31, 34, and 36, JJ31 and 51) by phosphorylation with cdk5/p26. cdk5/p26 showed similar enzymatic properties to p34cdc2/cyclin B kinase; the substrate specificity and inhibition by a p34cdc2 kinase specific inhibitor, butyrolactone I. However, p34cdc2/cyclin B kinase was distinguished from cdk5/p26 by its binding to p13suc1 protein and by its reactivity to anti-p34cdc2 antibodies. In spite of similar enzymatic properties of cdk5/p26 and p34cdc2/cyclin B kinase, cdk5/p26 did not display M-phase promoting activity when assayed with a cell-free system of Xenopus egg extract.

4-Butyrolactone↗

PubMed1995

Complete nucleotide sequences and the characteristics of two hepatitis B virus mutants causing serologically negative acute or chronic hepatitis B.

Hepatitis B virus (HBV) DNA was amplified by the polymerase chain reaction from the sera of a patient with acute hepatitis and a patient with chronic hepatitis. Both patients were negative for serum hepatitis B surface antigen and hepatitis B core antibodies and had been previously diagnosed as non-A, non-B, non-C, non-D, non-E hepatitis. The nucleotide sequence revealed an 8-nucleotide deletion in the X-gene coding region creating a C-terminally truncated X protein, and probable mutation of the enhancer II/core promoter element. In addition, DR2 showed a T-to-C mutation at the extreme 5'-terminus. These mutations within the X-gene coding region must suppress replication and expression of HBV DNA, and this seems to be responsible for absence of serological markers despite the presence of HBV infection.

Acute Disease↗

PubMed1995

Nucleotide sequence of hepatitis B virus isolated from subjects without serum anti-hepatitis B core antibody.

The nucleotide sequences of the precore/core and X open reading frames (ORFs) of hepatitis B virus (HBV) were studied in four subjects who were serologically negative for anti-hepatitis B core antibody. These subjects were positive for serum hepatitis B surface antigen and were considered to be asymptomatic HBV carriers. Sequencing of the precore/core ORF revealed precore wild type and 3 to 8 nucleotide substitutions (replacing 0 to 2 amino acids) in the core region compared with the sequence of subtype adr. These substitutions were not considered to have changed the epitope of the core antigen, resulting in the absence of anti-HBc as determined by a conventional diagnostic kit. The X ORF showed 1 to 5 nucleotide substitutions (replacing 1 to 3 amino acids) and the structure of the X protein and the core promoter/enhancer II complex appeared to be conserved. These findings strongly suggest that the absence of serum anti-HBc is not due to mutation of the HBV DNA but to an aberrant immune reaction of the host to HBV.

Adult↗

PubMed1995

Probable implication of mutations of the X open reading frame in the onset of fulminant hepatitis B.

A pathogenic role of precore-defective mutation in the onset of fulminant hepatitis B has been suggested. However, precore-defective mutants do not always cause fulminant hepatitis B and are not always isolated from affected patients. These findings strongly suggest the presence of some additional important mutations outside the precore region in fulminant hepatitis. In the present investigation an attempt was made to sequence the X open reading frame of hepatitis B virus DNA isolated from seven patients with fulminant hepatitis B and five patients with acute hepatitis B. The latter were used as controls. Since the X open reading frame encodes the X protein and contains the core promoter/enhancer II complex, some critical mutations may enhance or disrupt the replication and expression of hepatitis B virus DNA leading to fulminant hepatitis. A C-to-T substitution was found at nucleotide (nt) 1655, an A-to-T substitution at nt 1764 and a G-to-A substitution at nt 1766 in 4, 5 and 5 patients, respectively, out of the seven with fulminant hepatitis. These substitutions were not recognized in the patients with acute hepatitis. These mutations might change the function of the X protein and core promoter/enhancer II complex. It is suggested, therefore, that these mutations, as well as the precore-defective mutation, may play an important role in the pathogenesis of fulminant hepatitis.

Acute Disease↗

PubMed1995

Preparation and evaluation of Eudragit gels. VIII. Rectal absorption of 5-fluorouracil from Eudispert hv gels in rats.

Rectal absorption of the hydrophilic 5-fluorouracil (5-FU) in rats was studied with Eudispert hv gels with or without fatty acids as the rectal bases. In the absence of fatty acids, absolute bioavailabilities of 5-FU for Eudispert hv hydrogel and xerogel preparations increased approximately 2.5 times compared with those of Witepsol H-15 and PEG 2000 suppositories. When n-capric acid or linolenic acid was used as an absorption enhancer, absolute bioavailabilities of 5-FU were, respectively, 25.5 and 30.9% for Witepsol H-15 and 64.4 and 66.1% for PEG 2000. Furthermore, the absolute bioavailabilities of 5-FU for Eudispert hv hydrogel with n-capric acid or linolenic acid were 95.6% and 81.7%. The addition of capric acid or linolenic acid to the hydrogel was a useful method for increasing 5-FU permeability through the rectal membranes. These results are consistent with the observation that the total amounts of 5-FU remaining in the lumenal contents of the rectum and that accumulated in the rectal tissue decreased in relation to the increase in the bioavailabilities. Thus, the Eudispert hv hydrogel containing 5-FU with capric acid may be a useful rectal preparation for increasing the maximum plasma level and improving the absolute bioavailability of 5-FU.

Acrylic Resins↗

PubMed1995

Long-term hypothermic preservation of cardiac myocytes isolated from the neonatal rat ventricle: a comparison of various crystalloid solutions.

In this study, the functional and biochemical effects of crystalloid solutions on immature cardiac myocytes incubated under hypothermic conditions were evaluated. Cardiac myocytes were isolated from neonatal rat ventricles and cultured for 4 days, following which 12.5 x 10(5) myocytes per flask were incubated at 4 degrees C for 3, 6, 12, and 18 h in five types of crystalloid solutions: lactated Ringer's (LR), St. Thomas' Hospital (ST), University of Wisconsin (UW), 5% glucose-based potassium (GK), and normal saline (NS). The levels of creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) in the solutions were measured after each hypothermic incubation, following which the myocytes were cultured for an additional 24 h at 37 degrees C to evaluate the recovery of the myocyte beating rate. In the LR, UW, and NS groups, the recovery ratios of the myocyte beating rate were over 95% of the control (the beating rate prior to hypothermic incubation) at 3 h, but decreased to 20.3, 15.1, and 0%, respectively, at 18 h. The ST and GK groups had significantly lower recovery ratios than the other three groups (72.9% and 63.4%, respectively) at 3 h. The release of CPK and LDH in the LR, UW, and NS groups was significantly suppressed compared to the ST and GK groups, with the greatest suppression observed in the LR group. Moreover, the ST and GK groups had the highest CPK and LDH levels, respectively. Thus, LR solution had the least cytotoxic effects, indicating that it could be the most suitable basic solution of the various cardioplegic or preservation solutions during the neonatal period.

Adenosine↗

PubMed1995

Induction of cyclooxygenase protein and stimulation of prostaglandin E2 release by epidermal growth factor in cultured guinea pig gastric mucous cells.

The present study was undertaken to investigate whether epidermal growth factor (EGF) could stimulate prostaglandin E2 release, and if so, by what mechanism EGF would exert such an effect in gastric mucosal cells. In cultured guinea pig gastric mucous cells, EGF dose-dependently stimulated prostaglandin E2 release, with maximal stimulation observed at 10 ng/ml. EGF stimulated an increase in cyclooxygenase activity, which was reduced by protein synthesis inhibitor, actinomycin D, and cycloheximide. EGF also stimulated the enzyme protein synthesis estimated by Western blot analysis, whereas EGF did not stimulate phospholipase A2 activity. These results suggest that such an effect of EGF of de novo synthesis of cyclooxygenase protein and prostaglandin E2 release may be involved at least in part in the mechanism of EGF-induced local regulation of gastric mucosal integrity.

Animals↗

PubMed1995

Hepatitis E probably contracted via a Chinese herbal medicine, demonstrated by nucleotide sequencing.

Hepatitis E is endemic in developing countries and may occur as imported hepatitis in industrialized countries. A 46-year-old Japanese man developed immunoserologically diagnosed acute hepatitis E in Japan 4 months after he had made a trip to China. He had bought a Chinese herbal medicine there, taking it occasionally until approximately 6 weeks prior to the onset of acute hepatitis. Nucleotide sequencing of the 3' terminal region of the viral cDNA amplified from the patient's serum by polymerase chain reaction revealed a high degree of homology (99.8% of 752 nucleotides) with the Chinese strain. Thus, the results of sequencing suggest that his hepatitis E was caused by infection with the Chinese strain, via the Chinese herbal medicine.

Acute Disease↗

PubMed1995

Clinical study of symbrachydactyly of the foot.

Seventeen patients with symbrachydactyly of the foot are described. Patient characteristics including sex, the side of the affected foot, age at first medical examination, and condition of the nails were recorded. Compared with the unaffected side, the lengths of the proximal phalangeal and metatarsal bones were significantly shorter. The abnormalities of the feet were classified into four types: typical axial, atypical axial, medial ray, and rudimentary. The anomaly progresses from hypoplasia of the central rays to a deformity of the great toe, but rarely involves the fifth toe. The fact that this differs from symbrachydactyly of the hand is of considerable interest.

Adolescent↗

PubMed1995

An anterograde-retrograde labeling study of the carotid sinus nerve of the Japanese monkey (Macaca fuscata).

The sites of origin and termination of efferent and afferent fibers in the carotid sinus nerve (CSN) were investigated in the Japanese monkey. After application of a mixture of horseradish peroxidase (HRP) and wheat germ aggulutinin-conjugated HRP to the central cut end of the CSN, sensory ganglion neurons were labeled in the jugular ganglion of the vagus nerve, as well as in the superior and petrosal ganglia of the glossopharyngeal nerve. Many sympathetic ganglion neurons were also labeled retrogradely in the superior cervical ganglion. In the brain, many labeled terminals were seen ipsilaterally in the lateral division of the nucleus of the solitary tract (NST). A few neuronal cell bodies were also labeled ipsilaterally in a reticular region dorsomedial to the caudal one-third of the facial nucleus. The results indicate that the CSN of the Japanese monkey is composed mainly of afferent fibers terminating in the NST, that the afferent fibers in the CSN originate not only from the superior and petrosal ganglia of the glossopharyngeal nerve but also from the jugular ganglion of the vagus nerve, and that efferent fibers contained in the CSN arise from the medullary reticular formation and superior cervical ganglion.

Animals↗

PubMed1995

Overview of iron metabolism.

Regulation of iron balance is of particular interest, especially iron absorption, cellular iron metabolism and transferrin-transferrin receptor in hematopoiesis. Recent advances in molecular and cell biology have helped to reveal the mysteries of cellular iron metabolism concerning mRNA encoding ferritin and transferrin receptor synthesis. The physiology of transferrin and transferrin receptor is applied in the evaluation of erythropoiesis, i.e., erythron transferrin uptake in ferrokinetics and measurement of serum transferrin receptor. In iron absorption, much of the key mechanism remains unknown. The importance of iron metabolism in human beings is discussed in traditional areas of iron deficiency and nutrition. Iron overload is a new clinical problem to be solved in hemochromatosis or in relation to ischemic heart disease.

Cells↗

PubMed1995

Some cases of human male infertility are explained by abnormal in vitro human sperm activation.

OBJECTIVE: To determine if a human sperm activation assay can be used to evaluate males exhibiting otherwise unexplained infertility. DESIGN: Sperm from age-matched fertile and idiopathic infertile males were assayed in the human sperm activation assay and the results were compared. A portion of the sperm from the idiopathic infertile males also was used in assisted reproductive technology (ART) attempts at pregnancy. PATIENTS: Idiopathic infertile couples who had extensive fertility testing with no identified problems that would explain their infertility. Fertile males that had fathered one or more children. MAIN OUTCOME MEASURES: Sperm nuclear decondensation-recondensation and DNA synthesis. Pregnancies resulting from ART using semen from a male whose sperm responded abnormally in the human sperm activation assay. RESULTS: Thirteen (22%) of 59 idiopathic infertile males produced sperm that responded abnormally in the human sperm activation assay. Only 1 (1.7%) of 59 fertile males produced sperm that responded abnormally in the human sperm activation assay. The percentage of abnormal responders in the patient group exhibiting unexplained infertility was significantly higher than in the fertile male group. No sperm samples that responded abnormally in the human sperm activation assay resulted in pregnancies when used in ART. CONCLUSIONS: The human sperm activation assay is a new and independent indicator for some cases of infertility that otherwise would be unexplained. The human sperm activation assay appears to have utility in determining a sperm sample's efficacy for fertilization in ART attempts at pregnancy.

Cell Nucleus↗