Biomedical subjects

T Yamauchi

Publications and source records attributed to T Yamauchi.

At least 145 records · Page 8Linked to original sources

PubMed1999

Characterization of three splice variants and genomic organization of the mouse BMAL1 gene.

The BMAL1 gene encodes a member of the basic helix-loop-helix/PER-ARNT-SIM (bHLH/PAS) family of transcription factors. It is a key regulator of circadian rhythms. Using sequence information from human BMAL1 (hBMAL1) cDNAs previously reported by our laboratory, we have isolated and characterized cDNAs encoding three splice variants of the mouse BMAL1 (mBMAL1) gene. Of the three splice variants, mBMAL1b extends for 1878 bp in the coding sequence, which is 91% identical to that of hBMAL1b; its deduced amino acid sequence is 626 residues long and is 98% identical to that of hBMAL1b, and sequence identities in the bHLH, PAS-A, and PAS-B regions are 98, 100, and 100%, respectively. mBMAL1b' arises from alternative usage of exon 2, which results in a 7-amino-acid insertion and alternative splice acceptor usage at the intron 9/exon 10 splice junction, which causes an alanine residue deletion. mBMAL1b' encodes 632 amino acids and contains the bHLH/PAS domains. mBMAL1g' is generated by alternative splice acceptor usage at the intron 6/exon 7 splice junction, which results in a 28-bp deletion adjacent to the 5' end of the PAS domain. Since the 28-bp deletion shifts the reading frame, mBMAL1g' is predicted to encode a product of only 222 amino acids that lacks the PAS domain. The tissue distributions of the three splice variants showed some variation. The variations in the tissue distributions and predicted amino acid sequences suggest that the three splice variants may have different functions. Direct sequencing of the genomic mBMAL1 clones indicated that the coding sequence of mBMAL1 spans 32 kb and includes 17 exons. An unusual exon/intron donor sequence was found in intron 14, which begins with GC at the 5' end. Comparison with the bHLH/PAS family genes revealed that the intron/exon splice pattern of mBMAL1 most closely matches that of the mAhr, which suggests that BMAL1 and Ahr belong to the same subclass and may be derived from a common primordial gene.

ARNTL Transcription Factors↗

PubMed1999

Protein phosphatase 1 is involved in the dissociation of Ca2+/calmodulin-dependent protein kinase II from postsynaptic densities.

Autophosphorylation-dependent translocation of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) to postsynaptic densities (PSDs) from cytosol may be a physiologically important process during synaptic activation. We investigated a protein phosphatase responsible for dephosphorylation of the kinase. CaM kinase II was shown to be targeted to two sites using the gel overlay method in two-dimensional gel electrophoresis. Protein phosphatase 1 (PP1) was identified to dephosphorylate CaM kinase II from its complex with PSDs using phosphatase inhibitors and activators, and purified phosphatases. The kinase was released from PSDs after its dephosphorylation by PP1.

Animals↗

PubMed1999

Structural organization of the human Elk1 gene and its processed pseudogene Elk2.

In the ets gene family of transcription factors, ELK1 belongs to the subfamily of Ternary Complex Factors (TCFs) which bind to the Serum Response Element (SRE) in conjunction with a dimer of Serum Response Factors (SRFs). The primary structure of the human Elk1 gene was determined by genomic cloning. The gene structure of Elk1 spans 15.2 kb and consists of seven exons and six introns. The coding sequence resides on exons 3, 4, 5, 6 and 7. Sequencing of cDNA clones isolated from human hippocampus library revealed that the second exon was often skipped by an alternative splicing event. All introns commenced with nucleotides GT at the 5' boundary and ended with nucleotides AG at the 3' boundary, in agreement with the proposed consensus sequence for intron spliced donor and acceptance sites. Sequence inspection of the 5'-flanking region revealed the absence of a 'TATA' box and the presence of putative cis-acting regulatory elements such as Sp1, GATA-1, CCAAT, and c-Myb. Moreover, the sequence analysis of Elk2 locus on 14q32.3 confirmed that Elk2 gene corresponds to a processed pseudogene of Elk1 which has been reported between alpha 1 gene (IGHA1) and pseudo gamma gene (IGHGP) of immunoglobulin heavy chain. Furthermore, the results of Southern analysis using DNAs from human-mouse hybrid cell lines carrying a part of 14q32 region revealed that there is another locus hybridizing to Elk1 cDNA on 14q32.2 --> qter region in addition to Elk2 locus between IGHA1 and IGHGP loci.

Alternative Splicing↗

PubMed1999

Spontaneous disappearance of temporo-frontal arachnoid cyst in a child.

We report a child with a large temporo-frontal arachnoid cyst which resolved spontaneously. There was no history of a head injury. The patient was a boy aged 1.6 years. Though a large head was pointed out (+2SD), no therapeutic intervention was made because the relationship of the head circumference and the cyst was not established. No change in cyst size was visualized on the follow-up CTs at the age of 2.5 years and 6 years. At the age of 7 years, the arachnoid cyst had completely disappeared on CT. In order not to overlook a minute change in cyst size, the volumetry of the cyst and the intracranial cavity was performed, using the Photoshop, Macintosh. Both the cyst volume and the volume ratio of the cyst to the intracranial cavity slightly decreased and then increased. It is speculated that the cyst spontaneously ruptured by factors such as extreme breath holding and crying on the presence of the higher intracystic tension which might become a factor to accelerate spontaneous rupture of the cyst. Since a number of paediatric cases of symptomatic arachnoid cysts in need of surgical intervention has been larger than that of adult cases, we can speculate that a large arachnoid cyst might spontaneously resolve more frequently than we had expected. This case demonstrates that the surgical treatment of asymptomatic arachnoid cyst in the middle cranial fossa is not necessarily indicated in children.

Arachnoid Cysts↗

PubMed1999

Bone mineral density in patients with ossification of the posterior longitudinal ligament in the cervical spine.

Bone mineral density (BMD) has not been clearly determined in patients with ossification of the posterior longitudinal ligament (OPLL) in the cervical spine. BMD in patients with OPLL was measured in the third vertebral body in the lateral projection and in the distal part of the radius in the anteroposterior projection using dual-energy X-ray absorptiometry (DXA). Patients with OPLL had significantly higher BMD than healthy controls in both the lumbar spine and radius. Observing BMD by gender and age group, high BMD was recognized especially in female patients over 60 years of age. Significantly increased BMD was observed in patients with ankylosing spinal hyperostosis (ASH) in addition to OPLL. These findings suggest that patients with OPLL may tend to develop systemic hyperostosis, leading to the pathological ectopic ossification observed in OPLL.

Absorptiometry, Photon↗

PubMed1999

Isolation and mapping of a polymorphic CA repeat sequence at the human VRK1 locus.

VRK1 is a novel human putative serine/threonine kinase, and is located on chromosome 14 at band q32 where an autosomal recessive congenital microphthalmia (CMIC) is mapped. We isolated a polymorphic dinucleotide CA repeat marker from a genomic clone containing the human VRK1 gene. This polymorphism will be useful in genetic studies of disorders localized at the 14q32 region, such as CMIC.

Base Sequence↗

PubMed1999

Assignment of the human poly(A) polymerase (PAP) gene to chromosome 14q32.1-q32.2 and isolation of a polymorphic CA repeat sequence.

We report the chromosomal localization of the gene for human poly(A) polymerase (PAP) and the characterization of a newly isolated CA repeat near the PAP locus. By fluorescence in situ hybridization and polymerase chain reaction (PCR)-based analysis with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel, this gene was mapped on the q32.1-q32.2 region of chromosome 14. From a genomic clone containing the human PAP locus, we have isolated a polymorphic dinucleotide (CA) sequence. High heterozygosity (0.81) makes this polymorphism a useful marker in the genetic study of disorders localized at the 14q32 region, such as autosomal recessive congenital microphthalmia (CMIC).

Chromosome Mapping↗

PubMed1999

Effect of PSC 833 on the cytotoxicity of idarubicin and idarubicinol in multidrug-resistant K562 cells.

We examined the effect of PSC 833, a non-immunosuppressive cyclosporin analogue, on the cytotoxicity, accumulation and retention of idarubicin (IDA) and its 13-dihydro metabolite, idarubicinol (IDAol). P-glycoprotein (PGP)-overexpressing multidrug-resistant K562/D1-9 cells were used for these studies. PSC 833 had no effect on the cytotoxicity, intracellular accumulation, or retention of IDA and IDAol in the parent K562 cells. However, intracellular accumulation of IDA and IDAol in K562/D1-9 cells after a 60-min incubation was restored by 0.4 microM PSC 833 to 104% and 116%, respectively, of the level in parent K562 cells. The retention of IDA and IDAol in K562/D1-9 cells was also restored by 0.4 microM PSC 833. Consequently, 0.4 microM PSC 833 increased the sensitivity of K562/D1-9 cells to IDA and IDAol. The resistance index (RI) of IDA decreased from 20-fold to 4.0-fold, and the RI of IDAol decreased from 104-fold to 1.5-fold. These results suggest that the combination of IDA and PSC 833 may be effective in reversing PGP-mediated multidrug resistance in leukemia cells.

Antibiotics, Antineoplastic↗

PubMed1999

PPAR gamma mediates high-fat diet-induced adipocyte hypertrophy and insulin resistance.

Agonist-induced activation of peroxisome proliferator-activated receptor gamma (PPAR gamma) is known to cause adipocyte differentiation and insulin sensitivity. The biological role of PPAR gamma was investigated by gene targeting. Homozygous PPAR gamma-deficient embryos died at 10.5-11.5 dpc due to placental dysfunction. Quite unexpectedly, heterozygous PPAR gamma-deficient mice were protected from the development of insulin resistance due to adipocyte hypertrophy under a high-fat diet. These phenotypes were abrogated by PPAR gamma agonist treatment. Heterozygous PPAR gamma-deficient mice showed overexpression and hypersecretion of leptin despite the smaller size of adipocytes and decreased fat mass, which may explain these phenotypes at least in part. This study reveals a hitherto unpredicted role for PPAR gamma in high-fat diet-induced obesity due to adipocyte hypertrophy and insulin resistance, which requires both alleles of PPAR gamma.

Adipocytes↗

PubMed1999

Influence of transfusion-transmitted virus infection on the clinical features and response to interferon therapy in Japanese patients with chronic hepatitis C.

Recently, the genome of a novel DNA virus, transfusion-transmitted virus (TTV), was cloned from the plasma of a blood donor who had an elevated aminotransferase level but no serological markers of known hepatitis viruses. In this study, we investigated the influence of TTV infection on the clinical features and response to interferon (IFN) therapy in patients with chronic hepatitis C. We studied 247 patients who had received a 16- or a 24-week course of IFN-alpha therapy. The serum of these patients was analysed for TTV DNA using a hemi-nested polymerase chain reaction and TTV was detected in 114 patients (46%). No significant differences were found with respect to clinical features (gender, age, liver-related biochemical tests, hepatitis C virus (HCV) genotype and serum HCV RNA levels) between the patients who were positive for TTV DNA and those who were negative for TTV DNA. The fibrosis score was higher in TTV-positive patients (2.1 +/- 1.1) than in TTV-negative patients (1.7 +/- 1.1, P = 0.023). The biochemical sustained-response rate was 25% in TTV-positive patients and 25% in TTV-negative patients (not significant). A sustained HCV clearance rate was achieved in 26% of TTV-positive patients and in 22% of TTV-negative patients (not significant). TTV DNA clearance after IFN therapy was observed in 36 of 69 patients (52%) for whom stored serum samples were available. The disappearance of TTV DNA had no effect on the biochemical response to IFN therapy. In conclusion, TTV co-infection is frequently observed in Japanese patients with chronic hepatitis C. In chronic hepatitis C, TTV does not modify the clinical features or the response to IFN.

Adolescent↗

PubMed1999

Healthcare system in Japan.

In Japan, each person is obligated to enroll in a national health insurance plan. Based on egalitarianism, the system is managed by the government and it finances almost the entire healthcare needs. However low the total healthcare expenditure in Japan is compared with other major countries, the statistics cause concern for the Japanese population for the future growth of its health care. Even though the Japanese healthcare system faces many problems at present, it provides everyone with a sense of security. The key to its success is the sense of and art of balance in healthcare policy making.

Aged↗

PubMed1999

Osteocutaneous radial forearm flap for hand reconstruction.

We treated 5 patients (4 men and 1 woman) with bone and soft tissue defects or total thumb loss using an osteocutaneous radial forearm flap. Their ages at the time of surgery ranged from 27 to 65 years (mean, 53 years). Preoperative conditions were traumatic loss of the thumb in 3 cases and severe injuries with soft tissue and bone defects in 2. The length of the donated radius ranged from 2 to 7 cm (mean, 4.7 cm). In 4 patients the radial forearm flap was transposed, including the lateral antebrachial cutaneous nerve as a sensory flap. Simultaneous iliac bone grafting to the donor site was also performed in 4 of the cases. The follow-up period ranged from 44 to 87 months (mean, 64 months). All flaps survived over their full extent. Radiographic bone union was attained after 2 to 3 months (mean, 2.5 months). Two-point discrimination over the 4 sensory flaps ranged from 8 to 15 mm (mean, 10.8 mm). No radius fractures occurred. No patients showed signs of vascular insufficiency from sacrifice of the radial artery. The radial forearm flap provides thin skin of good texture together with bone for a 1-stage reconstruction. For patients with skin and bone defects of the hand, a radial forearm osteocutaneous flap is recommended. This flap is also recommended for patients with traumatic thumb loss, if the dorsalis pedis artery is absent, or if the patient declines using tissue from the foot.

Adult↗

PubMed1999

Long-term administration of adrenocorticotropin modulates the expression of IGF-I and TGF-beta 1 mRNAs in the rat adrenal cortex.

The effects of long-term adrenocorticotropin (ACTH) therapy on the expression of IGF-I and TGF-beta 1 on rat adrenal cortex was investigated. ACTH (0.1 mg/kg/day) or saline as control was injected intraperitoneally in 5-week-old Wistar rats every day for 4 weeks. ACTH significantly increased adrenal weight (P < 0.05) and serum corticosterone (P < 0.05). Competitive RT-PCR analysis on the adrenocortical mRNA showed increased IGF-I (P < 0.01) at 4 weeks of ACTH and increased TGF-beta 1 (P < 0.01) at 1 week of ACTH compared the control group. ACTH also significantly increased proliferating cell nuclear antigen mRNA level (P < 0.01), at 4 weeks of treatment, which correlated with IGF-I level (P < 0.01), but correlated negatively with ACTH-stimulated TGF-beta 1 level (P < 0.05). There was a weak correlation between IGF-I and serum corticosterone (P < 0.05), and between TGF-beta 1 mRNA levels and serum corticosterone concentration (P < 0.05). Histologically, ACTH induced hypertrophy in the zona fasciculata cells and increased the clear cells containing lipid deposits. Immunohistochemistry showed that IGF-I peptide was mainly expressed in the periphery of the zona fasciculata at 4 weeks of ACTH therapy, while the same therapy caused a slight increase in TGF-beta 1 expression in the same area. Our results show that an increase in adrenocortical growth resulting from ACTH treatment is associated with an increase in IGF-I mRNA expression but only a transient increase in TGF-beta 1 mRNA expression.

Adrenal Cortex↗

PubMed1999

Free vascularized fibula grafts in surgery of the upper limb.

Twenty patients with intractable diseases in the upper extremity were treated using free vascularized fibula grafts. There were 13 men and seven women. Three patients had traumatic bone defects, five had post-traumatic nonunions, two had congenital pseudoarthroses, seven had defects after tumor resection, and three had other lesions. The reconstructed sites were the humerus in two patients, the radius and/or ulna in 17, and the metacarpal and phalangeal bones in one. The length of the bone defect ranged from 3 to 18 cm (mean: 8.4 cm). Follow-up periods ranged from 6 to 204 months. No patient required additional bone grafts. The mean period required to obtain radiographic bone union was 4.4 months. There were no cases with fractures of the grafted bone, but malunion occurred in four cases. The vascularized fibula graft is indicated in patients with large bone defects or intractable nonunions in the humerus, radius, and/or ulna.

Adolescent↗

PubMed1999

Hydraulic analysis of the functional properties of saline-jet aspiration thrombectomy catheters.

The purpose of this paper is to clarify the functional properties of a saline-jet aspiration thrombectomy catheter based on hydraulics. This catheter, developed by the authors, aspirates intravascular thrombi by means of the effect of a high-speed jet. We have derived the relationships between Qs, Qa, and p(n), where Qs is the water supply rate, Qa is the water aspiration rate and p(n) is the negative pressure created in the distal lumen of the catheter. The relationships are represented by simple equations using c1 and c2, where c1 and c2 are coefficients proper to the catheter. We have obtained c1 and c2 experimentally for three types of catheter with different distal shapes. Two practical conclusions are derived from this study. First, making a short taper at the distal end increases the negative pressure created and is advantageous for thrombus aspiration. Second, making the diameter of the inner tube and nozzle smaller is essential for improving the aspiration ability of the catheter.

Catheterization↗

PubMed1999

Sevoflurane anaesthesia causes a transient decrease in aquaporin-2 and impairment of urine concentration.

Sevoflurane anaesthesia is occasionally associated with polyuria, but the exact mechanism of this phenomenon has not been clarified. Aquaporin-2 (AQP2) is an arginine vasopressin (AVP)-regulated water channel protein localized to the apical region of renal collecting duct cells and is involved in the regulation of water permeability. To elucidate the effect of sevoflurane anaesthesia on urine concentration and AQP2, we have compared serum and urinary concentrations of AVP, AQP2 and osmolar changes during sevoflurane and propofol anaesthesia. General anaesthesia was induced with sevoflurane or propofol in 30 patients for a variety of major surgical procedures. Blood and urine samples were obtained from patients at baseline, and 90 and 180 min after induction of anaesthesia. AVP and AQP2 concentrations were measured by radioimmunoassay. In both groups, plasma and urinary concentrations of AVP increased similarly during anaesthesia although plasma osmolality remained unchanged. Although urinary AQP2 excretion in the propofol group increased together with changes in plasma and urinary AVP, urinary AQP2 was significantly lower at 90 min in the sevoflurane group. Urine osmolality in the sevoflurane group also showed a transient but significant decrease in parallel with suppression of AQP2. Our data suggest that sevoflurane anaesthesia transiently produced an impaired AQP2 response to an increase in intrinsic AVP.

Adolescent↗

PubMed1999

The effects of intrinsic vasopressin on urinary aquaporin-2 excretion and urine osmolality during surgery under general anesthesia.

UNLABELLED: A radioimmunoassay has been established to measure urinary aquaporin-2 excretion (u-AQP2). To elucidate how u-AQP2 changes when endogenous vasopressin is increased independently of plasma osmolality, we estimated u-AQP2 during general anesthesia for surgery. We collected urine and blood samples from 50 patients before and 90 and 180 min after anesthetic induction. Plasma (29.1+/-12.6 pg/mL) and urinary (565.1+/-207.0 ng/gCr) vasopressin levels were markedly increased after anesthetic induction. Although no significant alteration of plasma osmolality or serum sodium concentration was observed during 180 min, u-AQP2 was significantly increased (preinduction 224.5+/-24.2 fmol/ mgCr; 90 min 243.3+/-31.8; 180 min 331.4+/-45.9), paralleling an increase of plasma and urinary vasopressin. The plasma vasopressin concentration after anesthetic induction was far in excess of that expected based on plasma osmolality. Individual plasma and urinary vasopressin concentrations correlated significantly with u-AQP2. At 180 min after anesthesia, plasma osmolality did not change, but urine osmolality decreased despite increased u-AQP2, and a preanesthetic positive correlation between urine osmolality and u-AQP2 disappeared. Thus, although u-AQP2 correlates with increased intrinsic vasopressin levels, the increase in u-AQP2 did not directly contribute to urine concentration. Apparently, an escape from the physiologic effects of high vasopressin level occurs during anesthesia via a mechanism independent of aquaporin-2. We conclude that the anesthetic would interfere with the urinary concentrating capacity at the level of AQP2-action. IMPLICATIONS: The excessive increase of intrinsic vasopressin exactly augmented urinary aquaporin-2 excretion, resulting in urine concentration; however, anesthesia seemed to modify this process possibly by interfering with the aquaporin-2 action.

Adolescent↗

PubMed1999

The role of serum imbibition for skin grafts.

Numerous studies of grafted skin suggest that full-thickness skin grafts are nourished by exudate from the recipient bed called a serum imbibition. However, whether serum imbibition by itself is sufficient for nourishment of skin grafts has not been shown definitely and directly. To clarify the role of serum imbibition, we performed a comparative study between 20 skin grafts and 20 musculocutaneous flaps. The nourishment of the cell in the skin graft is by serum imbibition. That in musculocutaneous flaps is mainly derived from blood supply. We evaluated the nourishment by means of the unique characteristics of the cell cycle. Once cells are put into a synthetic phase, they cannot reverse or stop the progress of the cell cycle. To take advantage of this characteristic of the cell cycle, prewounding methods (40 flaps were lifted once and put back to the original sites prior to the evaluation) were intended for the cells in pre-elevated skin to turn into a proliferating phase. Cells were examined by antibody against proliferating cell nuclear antigen immunohistologically, to determine whether they had turned into the proliferating phase or not. After 3 days, all flaps were reelevated; half (20 flaps) had their muscle layer and the neurovascular bundle removed to make a full-thickness skin graft. The rest (20 flaps) were only lifted. They were sutured back to the original sites. Ten skin grafts and musculocutaneous flaps each were harvested at 3 hours (1st day) and at 11 days (11th day) after the second operation. Bromodeoxyuridine, which is a thymidine analog and is taken into the cells in the synthetic phase, was introduced intraperitoneally 2 hours before the harvest. All flaps and grafts were evaluated histologically and immunohistologically. Proliferating cell nuclear antigen analysis showed that the prewounding method induced the cells of skin grafts and musculocutaneous flaps to proliferate before the implantation. Regarding the bromodeoxyuridine uptake, no significant differences could be seen between skin grafts and musculocutaneous flaps irrespective of their different nourishment. No structural changes, such as degenerative or necrotic, could be seen at the hair follicle and other glands even at the 11th day. Almost all of the layers of skin grafts survived as long as they were checked by light microscopy (hematoxylin and eosin stain). No differences could be seen between musculocutaneous flaps and skin grafts or between the 1st and 11th days in this study. We concluded that serum imbibition is sufficient for nourishment of skin grafts, just as blood supply is sufficient for nourishment of musculocutaneous flaps.

Animals↗