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Biomedical subjects

T Horiuchi

Publications and source records attributed to T Horiuchi.

At least 55 records · Page 3Linked to original sources
PubMed2001

Identification of DNA cis elements essential for expansion of ribosomal DNA repeats in Saccharomyces cerevisiae.

Saccharomyces cerevisiae carries approximately 150 ribosomal DNA (rDNA) copies in tandem repeats. Each repeat consists of the 35S rRNA gene, the NTS1 spacer, the 5S rRNA gene, and the NTS2 spacer. The FOB1 gene was previously shown to be required for replication fork block (RFB) activity at the RFB site in NTS1, for recombination hot spot (HOT1) activity, and for rDNA repeat expansion and contraction. We have constructed a strain in which the majority of rDNA repeats are deleted, leaving two copies of rDNA covering the 5S-NTS2-35S region and a single intact NTS1, and whose growth is supported by a helper plasmid carrying, in addition to the 5S rRNA gene, the 35S rRNA coding region fused to the GAL7 promoter. This strain carries a fob1 mutation, and an extensive expansion of chromosomal rDNA repeats was demonstrated by introducing the missing FOB1 gene by transformation. Mutational analysis using this system showed that not only the RFB site but also the adjacent approximately 400-bp region in NTS1 (together called the EXP region) are required for the FOB1-dependent repeat expansion. This approximately 400-bp DNA element is not required for the RFB activity or the HOT1 activity and therefore defines a function unique to rDNA repeat expansion (and presumably contraction) separate from HOT1 and RFB activities.

DNA Replication↗
PubMed2001

Yeast RNA polymerase I enhancer is dispensable for transcription of the chromosomal rRNA gene and cell growth, and its apparent transcription enhancement from ectopic promoters requires Fob1 protein.

At the end of the 35S rRNA gene within ribosomal DNA (rDNA) repeats in Saccharomyces cerevisiae lies an enhancer that has been shown to greatly stimulate rDNA transcription in ectopic reporter systems. We found, however, that the enhancer is not necessary for normal levels of rRNA synthesis from chromosomal rDNA or for cell growth. Yeast strains which have the entire enhancer from rDNA deleted did not show any defects in growth or rRNA synthesis. We found that the stimulatory activity of the enhancer for ectopic reporters is not observed in cells with disrupted nucleolar structures, suggesting that reporter genes are in general poorly accessible to RNA polymerase I (Pol I) machinery in the nucleolus and that the enhancer improves accessibility. We also found that a fob1 mutation abolishes transcription from the enhancer-dependent rDNA promoter integrated at the HIS4 locus without any effect on transcription from chromosomal rDNA. FOB1 is required for recombination hot spot (HOT1) activity, which also requires the enhancer region, and for recombination within rDNA repeats. We suggest that Fob1 protein stimulates interactions between rDNA repeats through the enhancer region, thus helping ectopic rDNA promoters to recruit the Pol I machinery normally present in the nucleolus.

Cell Division↗
PubMed2001

Difference in B cell activation between dermatomyositis and polymyositis: analysis of the expression of RP105 on peripheral blood B cells.

BACKGROUND: It has previously been shown that RP105, a new B cell surface protein, is lost in activated human B cells. OBJECTIVE: To investigate whether there is a difference in B cell activation between patients with dermatomyositis (DM) and those with polymyositis (PM) using RP105 as a marker. METHODS: The population of RP105 negative B cells (activated B cells) in the peripheral blood mononuclear cells of seven patients with dermatomyositis (DM) and 11 with polymyositis (PM) was analysed by flow cytometry. RESULTS: The percentage of RP105 negative B cells in the peripheral blood of patients with PM was low (5.8 (SD 2.4)%), similar to that of normal subjects. In contrast, all patients with DM showed increased RP105 negative B cell populations (33.0 (6.9)%). Bronchoalveolar lavage fluid from a patient with DM and active interstitial pneumonitis contained a large number of RP105 negative B cells. CONCLUSION: These findings suggest that the expansion of RP105 negative B cells is a hallmark of DM, and that B cell activation in DM may be pathogenetically different from that in PM.

Adult↗
PubMed2001

Analysis of purine- and pyrimidine-induced vascular responses in the isolated rat cerebral arteriole.

Effects of extraluminal UTP were studied and compared with vascular responses to ATP and its analogs in rat cerebral-penetrating arterioles. UTP, UDP, 2-methylthio-ATP, and alpha,beta-methylene-ATP dilated arterioles at the lowest concentration and constricted them at high concentrations. Low concentrations of ATP dilated the vessels; high concentrations caused a biphasic response, with transient constriction followed by dilation. Endothelial impairment inhibited ATP- and UTP-mediated dilation and potentiated constriction to UTP but not to ATP. ATP- and 2-methylthio-ATP- but not UTP-mediated constrictions were inhibited by desensitization with 10(-6) M alpha,beta-methylene-ATP or 3 x 10(-6) M pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). PPADS at 10(-4) M abolished the UTP-mediated constriction and induced vasodilation in a dose-dependent manner but did not affect the dilation to ATP. These results suggest that in rat cerebral microvessels 1) ATP and 2-methylthio-ATP induce transient constriction via smooth muscle P(2X1) receptors in the cerebral arteriole, 2) UTP stimulates two different classes of P(2Y) receptors, resulting in constriction (smooth muscle P(2Y4)) and dilation (possibly endothelial P(2Y2)), and 3) ATP and UTP produce dilation by stimulation of a single receptor (P(2Y2)).

Adenosine Triphosphate↗
PubMed2001

Role of potassium channels in regulation of brain arteriolar tone: comparison of cerebrum versus brain stem.

BACKGROUND AND PURPOSE: Potassium channels are important regulators of resting tone in large cerebral arteries, but their activity and distribution may vary according to vessel location and species studied. In the cerebral microcirculation in vivo, however, these channels appear to be silent at rest. Our goal was to determine the activity of potassium channels of brain arterioles from 2 origins under basal conditions in vitro. METHODS: Penetrating cerebral (40. 9+/-2.2 microm control diameter) and brain stem (36.2+/-1.2 microm) arterioles of rats were prepared from middle cerebral and basilar arteries, respectively. The internal diameter of cannulated and pressurized vessel was monitored with the inverted microscope before and after administration of potassium channel inhibitors. In addition, we studied the effect of nitric oxide synthase inhibition on potassium channel activity. RESULTS: Cerebral and brain stem arterioles were significantly constricted by 4-aminopyridine and low concentration of BaCl(2) but not by glibenclamide. The addition of N:(omega)-nitro-L-arginine to 4-aminopyridine further decreased diameters of both arterioles. Tetraethylammonium ion caused a significant constriction of brain stem but not cerebral arteriole. The brain stem arteriole was further constricted by additional N:(omega)-nitro-L-arginine. CONCLUSIONS: Voltage-dependent and inward-rectifier, but not ATP-sensitive, potassium channels are active under basal conditions of rat cerebral and brain stem arterioles. There is a regional difference in the activity of calcium-activated potassium channels, which, at rest, are open in brain stem but silent in cerebral arterioles. In addition, basal endogenous nitric oxide may not contribute to the activation of voltage-dependent and calcium-activated potassium channels.

4-Aminopyridine↗
PubMed2001

Involvement of prostaglandin E(2) in interleukin-1alpha-induced parathyroid hormone-related peptide production in synovial fibroblasts of patients with rheumatoid arthritis.

Synovial fibroblasts, established in culture from patients with RA, were treated with proinflammatory cytokines and prostaglandin E(2) (PGE(2)) for 24 h. These cells enhanced the production and the messenger RNA expression of PTH-related peptide (PTHrP) using proinflammatory cytokines, such as interleukin (IL)-1alpha, tumor necrosis factor-alpha without the coordination of other cytokines. In addition, PGE(2) which has been induced with IL-1, also enhanced the production of PTHrP. The IL-1alpha-induced PTHrP production was inhibited by PG H synthetase (Cox) inhibitors, indomethacin, and also by Cox-2 inhibitor, NS398. The synovial fibroblasts expressed PGE(2) receptor subtypes, EP2, EP3, EP4, but not EP1, as detected by RT-PCR. Of the PGE(2) receptor agonists, EP4 agonist showed the most marked induction of PTHrP, and EP2 agonist partly induced the production. However, these PGE(2) receptors were not induced by the treatment with IL-1alpha and PGE(2). These results suggest that induction of PGE(2) by IL-1alpha may be an important component of the PTHrP production of the inflammatory process in synovial tissues from patients with RA. These findings are the first to demonstrate that PGE(2) stimulates PTHrP production, which is mediated mostly by EP2 and EP4 receptors.

Arthritis, Rheumatoid↗
PubMed2001

Birth weight and gestation length of Japanese black calves following transfer of embryos produced in vitro with or without co-culture.

Birth weight and gestation length of calves following the transfer of in vitro produced (IVP) embryos with or without co-culture of cumulus cells, were compared to those produced in vivo (IVD). Spermatozoa from one Japanese Black bull were used for both IVP and IVD. IVP embryos were produced using two types of culture method: 1) co-culturing with cumulus cells in TCM 199 supplemented with calf serum (IVP-Co), and 2) non-co-culturing without cumulus cells in CR1aa supplemented with BSA / calf serum (IVP-NON-Co). Both IVP and IVD embryos were transferred non-surgically to Holstein recipients on day 7+/-1 of the estrous cycle. Birth weight and gestation length of half-sib single calves were analyzed. No differences were observed in birth weight and gestation length between IVP-Co and IVP-NON-Co calves (31.0 kg and 31.8 kg, and 291.9 days and 291.0 days, respectively). However, the birth weight of the IVP-Co and IVP-NON-Co calves was significantly higher than that of the IVD calves (P<0.01). Gestation length of the IVP-Co and IVP-NON-Co calves was also significantly longer than that of the IVD calves (P<0.01).

Animals↗
PubMed2001

Vestibuloplasty after secondary alveolar bone grafting.

OBJECTIVE: This paper introduces a surgical technique for vestibuloplasty after secondary alveolar bone grafting of patients with cleft lip and palate (CLP). This paper also reports on the patients who underwent this modified vestibuloplasty. SURGICAL PROCEDURE: The vestibuloplasty technique described in this paper consists of: (1) reduction of submucosal scar tissue of the upper lip, (2) V-Y plasty of the superficial mucosa, (3) placement of horizontal mattress sutures between nostril floor skin and freed marginal mucosa, (4) application of artificial skin to cover the exposed periosteal surface, and (5) use of a removable retention splint. CONCLUSION: This surgical procedure appears to be very useful for patients with CLP. The technique enables the surgeon to obtain an adequate sulcus depth around the graft area. In addition, this technique releases the mucosal scar contraction and improves the shape and mobility of the upper lip.

Adolescent↗
PubMed2001

Defect of lck in a patient with common variable immunodeficiency.

Common variable immunodeficiency (CVID) is a congenital immunological disorder characterized by defective antibody production with normal count of peripheral B lymphocytes. The basic immunologic defects that leads to CVID are still unknown, however, a proportion of CVID is suggested to be caused by decreased CD4+ helper T cell activity. In addition, recent reports indicate that a defect of T cell receptor (TCR)-associated signaling molecules results in congenital immune deficiency in human. In the present study, we investigated lck, a signaling molecule downstream of TCR, in a patient with CVID plus CD4 lymphopenia, and found an aberrantly spliced lck transcript lacking the entire exon 7 associated with the decrease in the expression of lck protein. An identical splicing abnormality has been previously demonstrated in a case of severe combined immunodeficiency with selective CD4 lymphopenia, although the case showed almost complete loss of the expression of lck protein. Considering these findings, the aberrant splicing of lck gene is suggested to be correlated, at least with a subset of congenital immunodeficiency plus CD4 lymphopenia.

3' Untranslated Regions↗
PubMed2001

[Self-reference effect on automatic and intentional memory].

The present study examined whether the self-reference effect occurred when memory retrieval was automatic. The process-dissociation procedure (e.g., Jacoby, 1991; Toth, Reingold, & Jacoby, 1994) was used to separate automatic and intentional, i.e., consciously controlled, components of memory in a word-stem completion task. In the learning phase, subjects were asked to rate trait words in one of the three ways of encoding: self-reference, semantic, and physical. Immediately after the phase, they performed an arithmetic problem for three minutes, and then were given the surprise word-stem recall task under an inclusion and exclusion performance conditions. Estimates derived from a process dissociation procedure showed the self-reference effect was found only in intentional memory, and not in automatic memory. Results therefore suggested that the self-reference effect to occur, intentional use of memory was necessary.

Adult↗
PubMed2001

[Effect of colforsin daropate hydrochloride after cardiopulmonary bypass].

We evaluated the effects of colforsin daropate hydrochloride (CDH) after cardiopulmonary bypass (CPB) in comparison with milrinone. CDH or milrinone were given during and after CPB combined with cathecholamines so as to maintain mean arterial pressure (mABP > 60 mmHg) and cardiac index (CI > 3.0 l.min-1.m-2). Hemodynamics measurement was done immediately after CPB (A 1), one hour after CPB (A 2) and after the chest closure (A 3). In CDH group, mABP was significantly higher compared with milrinone group. Also, CDH group showed a significant reduction in the cases of combined use of dobutamine (at A 2, A 3) and norepinephrine (at A 1, A 2) compared with milrinone group. In conclusion, colforsin daropate hydrochloride exerts more inotropic effect and could reduce the necessity of combined use of cathecholamine compared with milrinone.

Aged↗
PubMed2001

[Assessment of anesthesia satisfaction using direct interviews at post-anesthesia clinic].

The authors studied 5,034 consecutive patients undergoing elective surgery. Preoperative, intraoperative, and postoperative variables were gathered and patient satisfaction was assessed using direct interviews at pre- and post-anesthesia clinic. Fifty-nine percent of the 4,717 responders showed satisfaction and 4% showed dissatisfaction with anesthesia. The most undesirable perioperative outcome was vomiting/nausea. Other undesirable outcomes include discomfort of urine catheter, sore throat, memory of extubation, postoperative pain and so on. Anesthesiologist can improve the quality of anesthesia by preoperative explanation and preventative management for undesirable perioperative outcomes.

Anesthesia↗
PubMed2001

[Anesthetic management of the King-Denborough syndrome].

The King-Denborough syndrome (KDS) is a congenital myopathy with musculoskeletal abnormalities, and definitely associated with susceptibility to malignant hyperthermia (MH). We present the first report in Japan concerning the management of a KDS patient. A 2-year-old boy was scheduled for cryptorchidopexy. He had some physical signs of KDS, e.g. pectus excavatum, low-set ears, malar hypoplasia, micrognathia, ptosis and down-slanting palpebral fissures. Moreover, his mother and maternal grand-uncle had medical history of MH. Therefore, he was diagnosed as KDS by pediatricians. All of the preoperative examinations, including serum creatine phosphokinase level, are normal. Previous 15 case-reports suggest that in KDS patients MH might be triggered by volatile anesthetics. To avoid the trigger, we maintained the general anesthesia with propofol, fentanyl and vecuronium. During perioperative period, his body temperature was 36.3-38.1 degrees C, and no symptom of MH was observed. It is rare that a diagnosis of KDS is made preoperatively since the MH-induction is involved in the criteria of this syndrome. In the anesthesia of patients who are suspected of KDS from their characteristic features or familial histories, an anesthesiologist should pay attention to prevent MH. The total intravenous anesthesia method appears useful for the management of KDS.

Abnormalities, Multiple↗
PubMed2001

[Ketamine lollipop for pediatric premedication].

Lollipop containing ketamine (50 mg) was evaluated for the premedication of pediatric patients. The subject of this study were 12 children aged from 1 year 7 months to 6 years. They received the lollipop and showed relatively good emotional state and no typical side effects. These results suggest good possibility of ketamine lollipop as the premedication for pediatric patients.

Administration, Oral↗
PubMed2001

[Propofol Concentration during Maintenance of and Emergence from Propofol/Epidural Anesthesia: Comparison between Younger and Elderly Patient].

To investigate the influence of patient age on the sensitivity to propofol, we measured blood propofol concentrations in ten elderly (over 70 years of age) and ten younger (under 60 years of age) patients undergoing elective abdominal surgery during propofol/epidural anesthesia. Bispectral index (BIS) was continuously recorded for monitoring anesthetic effect, and the infusion rate of propofol was controlled to keep BIS at 50 after the induction of anesthesia with propofol. At steady-state before skin incision, propofol concentrations to maintain BIS at 50 in younger and elderly patients were 4.3 +/- 1.6 micrograms.ml-1 and 3.2 +/- 1.3 micrograms.ml-1, respectively, and there was no significant difference. Mean blood propofol concentrations were 4.0 +/- 1.5 micrograms.ml-1 for the younger group and 3.2 +/- 1.7 micrograms.ml-1 for the elderly group at the end of surgery immediately before discontinuation of propofol, and they were 1.9 +/- 0.7 micrograms.ml-1 (BIS = 86 +/- 7) for the younger group and 1.5 +/- 0.8 micrograms.ml-1 (BIS = 84 +/- 3) for the elderly group at the time of eye opening with no significant difference. We found appreciable individual variations in the propofol concentrations both in younger and elderly patients in this study.

Abdomen↗
PubMed2001

[Relationship between minimum alveolar concentration and electroencephalographic bispectral index as well as spectral edge frequency 95 during isoflurane/epidural or sevoflurane/epidural anesthesia].

To investigate the relationship between minimum alveolar concentration (MAC) and electroencephalographic variables, we measured the bispectral index (BIS) and the spectral edge frequency 95 (SEF 95) in 17 patients undergoing elective surgery during isoflurane/epidural (n = 8) or sevoflurane/epidural (n = 9) anesthesia. Patients received 2.0 MAC end-tidal concentrations of isoflurane or sevoflurane, and the BIS and the SEF 95 were recorded after 15 min of an unchanged end-tidal concentration. The concentration of the inhalational agent was decreased to 1.2 MAC, and measurements were repeated again. During isoflurane anesthesia, the BIS increased significantly (3.6 +/- 3.9 at 2.0 MAC, 43.5 +/- 9.2 at 1.2 MAC [mean +/- SD]). In contrast, the BIS did not change significantly during sevoflurane anesthesia (35.3 +/- 8.4 at 2.0 MAC, 42.8 +/- 6.1 at 1.2 MAC). There were significant differences in the BIS and the SEF 95 at 2.0 MAC between isoflurane and sevoflurane groups. In contrast, the BIS and the SEF 95 showed no difference at 1.2 MAC between the groups. These findings suggest that different inhalational anesthetics may have different effects on the BIS and the SEF 95.

Adult↗