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Biomedical subjects

K Aoki

Publications and source records attributed to K Aoki.

At least 235 records · Page 13Linked to original sources
PubMed1999

HPLC profile analysis of hepatoprotective oleanene-glucuronides in Puerariae Flos.

In order to confirm the constitution of hepatoprotective oleanene glucuronide (OG), HPLC profile analyses of the total OG fractions of both Puerariae Thomsonii Flos (the flowers of Pueraria thomsonii) and Puerariae Lobatae Flos (the flowers of P. lobata) were performed. No remarkable difference in the HPLC profiles with respect to OGs in the flowers was shown, in contrast to those of the roots. By repeated chromatography of the total OG fraction of Puerariae Thomsonii Flos, soyasaponin I (1), kaikasaponin III (2) and kakkasaponin I (3), which had been already isolated from Puerariae Lobatae Flos, were obtained. The hepatoprotective activity of 2 towards immunologically induced liver injury was significantly more effective than that of 1. This information supported previously obtained structure-hepatoprotective relationship data which was measured on another model. The structure-activity relationship information which suggested that the hydroxymethyl group of the galactosyl unit would enhance the hepatoprotective activity was also substantiated.

Animals↗
PubMed1999

Cloning of a gene encoding hydroxyquinol 1,2-dioxygenase that catalyzes both intradiol and extradiol ring cleavage of catechol.

Two Escherichia coli transformants with catechol 1,2-dioxygenase activity were selected from a gene library of the benzamide-assimilating bacterium Arthrobacter species strain BA-5-17, which produces four catechol 1,2-dioxygenase isozymes. A DNA fragment isolated from one transformant contained a complete open reading frame (ORF). The deduced amino acid sequence of the ORF shared high identity with hydroxyquinol 1,2-dioxygenase. An enzyme expressed by the ORF was purified to homogeneity and characterized. When hydroxyquinol was used as a substrate, the purified enzyme showed 6.8-fold activity of that for catechol. On the basis of the sequence identity and substrate specificity of the enzyme, we concluded that the ORF encoded hydroxyquinol 1,2-dioxygenase. When catechol was used as a substrate, cis,cis-muconic acid and 2-hydroxymuconic 6-semialdehyde, which were products by the intradiol and extradiol ring cleavage activities, respectively, were produced. These results showed that the hydroxyquinol 1,2-dioxygenase reported here was a novel dioxygenase that catalyzed both the intradiol and extradiol cleavage of catechol.

Amino Acid Sequence↗
PubMed1999

Effects of papain on isolation of single smooth muscle cells from the guinea pig longitudinal ileum.

The methods for isolation of single cells from the guinea pig longitudinal ileum were investigated with focussing on the papain concentration for the digestion of the ileum. The ileal muscle was minced. The minced muscle was loaded with fluorescent probes of calcein or fura 2, and treated with papain for 30 min at various concentrations. Papain at concentrations more than 1 U/ml reduced both calcein fluorescence and fura-2-signal evoked by carbachol. Carbachol-induced fura-2-signal was more sensitive to papain than calcein fluorescence, suggesting that proteins related to the formation of receptors are more vulnerable than membrane lipids or proteins limiting the membrane permeability upon the exposure to papain. The resultant yield of single cells was highest at 0.56 U/ml of papain without affecting calcein and fura-2 fluorescence responses, thus this concentration appeared to be appropriate for the isolation of single cells from the ileum. Single cells alive contracted dose dependently by the exposure to carbachol (0.1-10 microM) under the microscopic measurement, and were appeared to grow confluent in culture for approximately 15 days. These results suggest that the low concentration, 0.56 U/ml, of papain in the isolation medium is better to obtain functional cells from the guinea pig ileum.

Animals↗
PubMed1999

Dehydroepiandrosterone suppresses the elevated hepatic glucose-6-phosphatase and fructose-1,6-bisphosphatase activities in C57BL/Ksj-db/db mice: comparison with troglitazone.

The effect of dehydroepiandrosterone (DHEA) on the hepatic and muscle glucose metabolizing enzymes and on blood glucose were investigated in insulin-resistant diabetic C57BL/KsJ-db/db mice and their heterozygote littermates (db/+m). The results were compared with those after troglitazone administration under the same conditions. Despite hyperinsulinemia, hepatic glucose-6-phosphatase (G6Pase) and fructose-1,6-bisphosphatase (FBPase) activities are higher in db/db than in db/+m mice. Dietary administration of DHEA and that of troglitazone for 15 days to respective groups of five mice each significantly decreased blood glucose in db/db mice and hepatic G6Pase and FBPase activities in both db/db and db/+m mice. Hepatic G6Pase and FBPase activities showed a linear relationship with blood glucose in all groups of mice, suggesting that the activities of G6Pase and FBPase are closely related to blood glucose levels. Because androstenedione, a DHEA metabolite, barely affected either of these enzyme activities or blood glucose in db/db mice, the actions of DHEA, which are similar to those of troglitazone, are presumed to be caused by DHEA itself. DHEA is considered to be a modulating agent for the activities of hepatic gluconeogenic enzymes in db/db mice.

Androstenedione↗
PubMed1999

[Pathophysiology and treatment results of auditory ossicle damage due to earpick-induced trauma].

We examined 10 ears from 10 patients undergoing ossiculoplasty due to a diagnosis of earpick-induced traumatic ossiculodialysis. Surgical findings induced dialysis of the incudostapedial joint in 5 patients, however, dialysis of this articulation was infrequent. Abnormal stapes were observed in 9 patients, including 6 patients with foot plate invagination. Five patients had a concomitant perilymphatic fistula. Perilymphatic fistula was found in 5 of 6 patients with vertigo but not in patients with tinnitus. It is thought to be difficult to diagnose ossiculodialysis from a tympanogram alone; stapedius reflex could possibly be referred to at diagnosis. The auditory ossicle was reconstructed using the stapes head and foot plate in patients with normal foot plates. In patients with ossicular invagination, the position of the auditory ossicle was reduced and the perilymphatic fistula closed, with ossicular reconstruction as needed. The surgical results were satisfactory, and auditory acuity was improved in 90% of the patients.

Adult↗
PubMed1999

[Eustachian tube function of adhesive tympanum--pre and post operative evaluation].

We examined the eustachian tube function of 48 patients with adhesive tympanum who had undergone operations. Preoperative tubal openings were observed and determined in 57% of these cases by the dynamic movement of the tympanic membrane, 35% by the impedance method, and 74% by the sonotubometry and was found to be lower than in healthy subject. Furthermore the tubal opening duration in sonotubometry was significantly shortened in patients with adhesive tympanum as compared to healthy subjects. However, in a few cases of adhesive tympanum, tubal opening was prolonged remarkably, and indicated a mixed tendency of stenotic and patulous types. Postoperative tubal opening was not seen in 90.9% of the cases as determined by the inflation-deflation test. Mucociliary function was poor in 81.3% of the cases. No remarkable differences were seen between cases of total adhesion and those of posterosuperior quadrant adhesion by any method. The condition of the tympanic membrane and the tympanic cavity after operation, and the eustachian tube function were not correlated in some cases and indicated possible involvement of other factors.

Adolescent↗
PubMed1999

[Trachoma].

Explore the source record for details and available documents.

Conjunctivitis, Inclusion↗
PubMed1999

Regulation of macrophage migration inhibitory factor (MIF) expression by glucose and insulin in adipocytes in vitro.

BACKGROUND: It has been reported that macrophage migration inhibitory factor (MIF) stimulated insulin secretion from pancreatic islet beta-cells in an autocrine manner, which suggests its pivotal role in the glucose metabolism. According to this finding, we evaluated MIF expression in cultured adipocytes and epididymal fat pads of obese and diabetic rats to investigate its role in adipose tissue. MATERIALS AND METHODS: The murine adipocyte cell line 3T3-L1 was used to examine MIF mRNA expression and production of MIF protein in response to various concentrations of glucose and insulin. Epididymal fat pads of Otsuka Long-Evans Tokushima fatty (OLETF) and Wistar fatty rats, animal models of obesity and diabetes, were subjected to Northern blot analysis to determine MIF mRNA levels. RESULTS: MIF mRNA of 3T3-L1 adipocytes was up-regulated by costimulation with glucose and insulin. Intracellular MIF content was significantly increased by stimulation, whereas its content in the culture medium was decreased. When the cells were treated with cytochalasin B, MIF secretion in the medium was increased. Pioglitazone significantly increased MIF content in the culture medium of 3T3-L1 cells. However, MIF mRNA expression of both epididymal fat pads of OLETF and Wistar fatty rats was down-regulated despite a high plasma glucose level. The plasma MIF level of Wistar fatty rats was significantly increased by treatment with pioglitazone. CONCLUSION: We show here that the intracellular glucose level is critical to determining the MIF mRNA level as well as its protein content in adipose tissue. MIF is known to play an important role in glucose metabolism as a positive regulator of insulin secretion. In this context, it is conceivable that MIF may affect the pathophysiology of obesity and diabetes.

3T3 Cells↗
PubMed1999

Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro.

BACKGROUND: Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system. MATERIALS AND METHODS: Full-length recombinant adenovirus DNA was generated in vitro by Cre-mediated recombination between loxP sites in a linearized shuttle plasmid containing a transgene and adenovirus genomic DNA. RESULTS: After transfection of Cre-treated DNA into 293 cells, replication-defective viral vectors were rapidly obtained without detectable wild-type virus. CONCLUSION: This system facilitates the development of recombinant adenoviral vectors for basic and clinical research.

Adenoviridae↗
PubMed1999

[Recent advances in the management of severely burned patients].

With recent advances in the systemic care of burns, patients with burns covering 80% of their body surface can frequently survive. The percentage of total body surface area burn for an expected 50% mortality rate has improved to 98% for children and 72% for adults in one burn center in the USA. From the results of 11 burn units in Tokyo, the mortality rate of burn patients with a prognostic burn index of 90-100 was 51.4%. The improvement is attributable to advances in the understanding of the pathophysiology of severe burns as a systemic inflammatory response syndrome. Improved cardiopulmonary management of extensive burns and respiratory tract burns has also played a part in reducing the mortality rate. Individualized fluid resuscitation programs based on hemodynamic monitoring have reduced the incidence of burn shock and acute renal failure. Early eschar excision and wound closure by immediate grafting have further reduced the mortality rate from extensive full-thickness burns. The use of bilayer artificial skin has improved the survival and cosmetic results of early eschar excision in patients with massive full-thickness burns. Cultured autologous epidermal sheets hold promise if used on an appropriate dermal bed.

Adolescent↗
PubMed1998

RP58 associates with condensed chromatin and mediates a sequence-specific transcriptional repression.

An approximately 120-amino acid domain present generally at the NH2 termini, termed the POZ domain, is highly conserved in various proteins with zinc finger DNA binding motifs. We have isolated a novel protein sharing homology with the POZ domain of a number of zinc finger proteins, including the human BCL-6 protein. By using a binding site selection technique (CAST), a high affinity binding site of the protein was determined to be (A/C)ACATCTG(G/T)(A/C), containing the E box core sequence motif. The protein was shown to repress transcription from a promoter linked to its target sequences and was hence named RP58 (Repressor Protein with a predicted molecular mass of 58 kDa). Immunogold electron microscopic study revealed that almost all RP58 is localized in condensed chromatin regions. These observations demonstrate for the first time that a protein mediating a sequence-specific transcriptional repression associates with highly condensed chromatin. We suggest that RP58 may be involved in a molecular link between sequence-specific transcriptional repression and the organization of chromosomes in the nucleus.

Amino Acid Sequence↗
PubMed1998

L-type calcium channels in the axon terminal of mouse bipolar cells.

Two types of calcium current (I(Ca)) have been identified in the bipolar cell of the mouse retina: a transient (T-) type current and a long lasting (L-) type current. It has been suggested that the former is present in the soma and the latter in the axon terminal. To establish the cellular localization of the two types of I(Ca), bipolar cells of the mouse retina was studied electrophysiologically in a slice preparation, and immunocytochemically by staining specific calcium channels in isolated cells. The dihydropyridine-sensitive L-type I(Ca) was recorded in the axon terminal, and the T-type current was recorded in the somatic region. Strong immunoreactivity to a polyclonal antibody against the L-type calcium channel was found in the axon terminal. These observations suggest that the L-type I(Ca) is generated at the axon terminal and contributes to the transmission of sustained depolarization.

Animals↗
PubMed1998

Complete sequence and gene organization of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3.

The complete sequence of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3, has been determined by assembling the sequences of the physical map-based contigs of fosmid clones and of long polymerase chain reaction (PCR) products which were used for gap-filling. The entire length of the genome was 1,738,505 bp. The authenticity of the entire genome sequence was supported by restriction analysis of long PCR products, which were directly amplified from the genomic DNA. As the potential protein-coding regions, a total of 2061 open reading frames (ORFs) were assigned, and by similarity search against public databases, 406 (19.7%) were related to genes with putative function and 453 (22.0%) to the sequences registered but with unknown function. The remaining 1202 ORFs (58.3%) did not show any significant similarity to the sequences in the databases. Sequence comparison among the assigned ORFs in the genome provided evidence that a considerable number of ORFs were generated by sequence duplication. By similarity search, 11 ORFs were assumed to contain the intein elements. The RNA genes identified were a single 16S-23S rRNA operon, two 5S rRNA genes and 46 tRNA genes including two with the intron structure. All the assigned ORFs and RNA coding regions occupied 91.25% of the whole genome. The data presented in this paper are available on the internet at http:@www.nite.go.jp.

Chromosomes, Archaeal↗