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Biomedical subjects

T Seki

Publications and source records attributed to T Seki.

At least 199 records · Page 11Linked to original sources

Expression of NOS, PSA-N-CAM and S100 protein in the granule cell migration pathway of the adult guinea pig forebrain.

To investigate the possible role of nitric oxide (NO) in adult neurogenesis and neuron-glial migration in the rostral migratory stream (RMS), we used a double-labeled immunofluorescence technique together with confocal laser scanning microscopy, and examined the localization of nitric oxide synthase (NOS), the highly polysialylated isoform of neural cell adhesion molecule (PSA-N-CAM), and the astroglial marker in brain, S100 protein (S100), throughout the length of the subependymal layer (SEL) to olfactory bulb (OB) pathway of the adult guinea pig forebrain. Blast-like, beaded, clustered immature cellular elements stained for PSA-N-CAM and those having a typical astrocytic phenotypes positive for S100 protein were densely interlaced throughout the entire length of the SEL. Some S100 positive ependymoglial cells (tanycytes) gave off their basal projections into the closely packed PSA-N-CAM immunopositive clusters in the rostral extension of the subependymal zone (SEZre). The SEL was devoid of NOS immunoreactivity. A dense network of punctate, fenestrated and radially oriented immature cellular elements positive both for NOS and PSA-N-CAM intermingled and overlapped in the inner part of the internal granular layer (IGr), whereas in the outer part, PSA-N-CAM expression gradually diminished and the cells shifted to mature bipolar, spherical or spindle-shaped granule cells with uniform cellular contours, which were exclusively immunopositive for NOS. Radially oriented astroglial phenotypes were intertwined with PSA-N-CAM neuronal clusters in the SEL, and were closely apposed to NOS neuronal elements in the IGr. In summary, these results showed a distinct separation of neurons and glia as revealed by PSA-N-CAM and S100 protein immunostaining, and an inverse spatio-temporal correlation of expression between PSA-N-CAM (immature neuroblasts) and NOS (mature neurons) in the adult guinea pig RMS.

Animals↗

Removal of polysialic acid-neural cell adhesion molecule induces aberrant mossy fiber innervation and ectopic synaptogenesis in the hippocampus.

The mossy fiber axons of both the developing and adult dentate gyrus express the highly polysialylated form of neural cell adhesion molecule (NCAM) as they innervate the proximal apical dendrites of pyramidal cells in the CA3 region of the hippocampus. The present study used polysialic acid (PSA)-deficient and NCAM mutant mice to evaluate the role of PSA in mossy fiber development. The results indicate that removal of PSA by either specific enzymatic degradation or mutation of the NCAM-180 isoform that carries PSA in the brain causes an aberrant and persistent innervation of the pyramidal cell layer by mossy fibers, including excessive collateral sprouting and/or defasciculation of these processes, as well as formation of ectopic mossy fiber synaptic boutons. These results are considered in terms of two possible effects of PSA removal: an increase in the number of mossy fibers that can grow into the pyramidal cell layer and an inhibition of process retraction by formation of stable junctions including synapses. As these defects on granule cells in the adult animal and PSA-positive granule cells continue to be produced in the mature brain, the present findings may be relevant to previous studies suggesting that PSA-NCAM function is required for long-term potentiation, long-term depression, and learning behaviors associated with hippocampus.

Animals↗

Isolation of a cDNA encoding mouse DNA topoisomerase III which is highly expressed at the mRNA level in the testis.

A cDNA (mTOP3) encoding mouse DNA topoisomerase III (topo III) was cloned using the human TOP3 cDNA as a probe. The deduced amino acid sequence of mouse topo III showed 86.8% identity with that of human topo III. Mouse TOP3 mRNA was highly expressed in the testis in comparison with other tissues. The TOP3 mRNA level in the testis increased slightly 14 days after birth and showed a marked increase in 17 days, times when the cells in pachytene phase begin to appear and increase in number.

Age Factors↗

Use of differential subtraction method to identify genes that characterize the phenotype of cultured rheumatoid arthritis synoviocytes.

OBJECTIVE: To identify the genes that characterize the distinctive phenotype of cultured rheumatoid arthritis (RA) fibroblastoid synoviocytes. METHODS: A representational difference method was used to subtract complementary DNA (cDNA) from cultured RA fibroblastoid synoviocytes with cDNA from noninflammatory osteoarthritis synoviocytes. The genes were identified by DNA sequencing, and their relative expression was determined by Northern blot analysis. RESULTS: Twenty-four genes were identified, including novel genes such as a human homolog of mouse semaphorin E and one homologous to N-acetylglucosamine-6-sulfatase. Eleven of these genes were constitutively overexpressed in the rheumatoid synoviocyte line, including a chemokine, stromal cell-derived factor 1, and several genes capable of mediating synoviocyte-leukocyte interactions, including vascular cell adhesion molecule 1 and Mac-2 binding protein. Three genes (lumican, biglycan, and insulin-like growth factor binding protein 5) encoded extracellular matrix components, suggesting that distinct stromal-synoviocyte interactions may be mediated by this phenotype. Two interferon-inducible genes of unknown function were also found, emphasizing the presence of activation-like features in the phenotype. CONCLUSION: A general method for the identification of differences in patterns of gene expression revealed that cultured RA fibroblastoid synoviocytes overexpress certain proinflammatory genes that are potentially relevant to lymphocyte and monocyte entry and interactions. The features of the genes identified in these mesenchymal cells suggest that they facilitate localization of immune reactions to the joint through leukocyte chemokinesis, cell-cell adhesion, and matrix specialization. The further characterization of these genes should help in resolving whether this phenotype is the consequence of modulation and imprinting by an inflammatory milieu or, more likely, whether it reflects the intrinsic lineage characteristics of intimal lining synoviocytes.

Amino Acid Sequence↗

LHRH neurons migrate into the trigeminal nerve when the developing olfactory nerve fibers are physically interrupted in chick embryos.

Most LHRH neurons actively migrate from the olfactory epithelium to the forebrain during embryonic days (ED) 3.5-8. When a small piece of the membrane filter was placed on the central course of the olfactory nerve in ED 3.5-5 chick embryos, LHRH neurons deviated from their regular migratory course at ED 6.5-7.5 to follow a route along the PSA-NCAM-positive medial and lateral nasal branches of the ophthalmic nerve of the trigeminal nerve. The olfactory nerve fibers which were specifically immunoreactive for somatostatin also deviated into the ophthalmic nerve. Enzymatic removal of PSA using endoneuraminidase did not interfere with the migration of LHRH neurons into the ophthalmic nerve bundle of the trigeminal nerve. The presence of structural supports seems to be primarily of importance in the migration of LHRH neurons along the olfactory and trigeminal nerve bundles. PSA may be less important for the migration of the LHRH neurons along peripheral neural elements.

Animals↗

Arg506Gln mutation of the coagulation factor V gene not detected in Japanese pulmonary thromboembolism.

The incidence of pulmonary thromboembolism (PTE) is lower in Japanese than in Caucasians. The basis for the different incidence has not been clarified. A poor anticoagulant response to activated protein C based on a single point mutation of the factor V gene (Arg506Gln) was found to be a pathogenetic factor for venous thrombosis and PTE in North America and Europe. We investigated whether the Arg506Gln mutation of factor V is responsible for the occurrence of PTE among Japanese. We analyzed genomic DNA prepared from fresh peripheral blood of 25 patients with PTE of unknown etiology (12 of acute type and 13 of chronic type) and that of 110 controls without respiratory or circulatory disorders. To detect the Arg506Gln mutation, 267 bp DNA fragments of the factor V gene including the Arg506Gln region were amplified by PCR, digested by MnlI and electrophoresed. After digestion of PCR products with MnlI, DNA fragments of 163 bp length, but not DNA fragments of 200 bp length, were identified in all samples, indicating the absence of the Arg506Gln mutation in the patients with PTE and control subjects. These results suggest that the Arg506Gln mutation is absent or very rare and not an important pathogenetic factor for PTE in Japanese.

Aged↗

Experience with kidney transplantation in children and adolescents.

BACKGROUND: Paediatric kidney transplantation has different aspects in adults in terms of underlying primary renal disease, surgical technique, perioperative care and graft prognosis. Significant urological problems are present in a high percentage of paediatric recipients. This study was undertaken to characterize paediatric kidney transplantation as performed at our institutes. METHODS: Twenty-eight patients (age range 4-17 years, 10 girls, 18 boys) were included in this study. We analyzed (1) urologic procedures performed prior to or simultaneously with transplantation, (2) intraoperative changes in haemodynamics, (3) postoperative complications and (4) acute allograft rejection and graft prognosis. RESULTS: Of 4 patients with lower urinary tract abnormalities (neurogenic bladder in 3, posterior urethral valve in 1), 2 underwent augmentation ileocystoplasty prior to transplantation. The Mitrofanoff procedure as a diversion for neourethra was also performed in 2 of the 4 patients. All these 4 patients were managed with clean intermittent catheterization. Central venous pressure changes before and after graft vessel declamping were much greater in patients with body weight below 25 kg than in those above 25 kg. Five surgical complications and 6 infections were encountered postoperatively and hypertension was lasting in 6. Thirteen patients experienced 19 periods of acute allograft rejection. All of the 24 patients in the cyclosporin era (1986-) overcame rejections and they are all alive with good graft function. CONCLUSIONS: As long as proper pretransplant patient evaluation and management, and intensive perioperative care are undertaken, good prognosis of renal allograft can be achieved in young patients.

Adolescent↗

Potential for qualitative diagnosis of tumors and tumorous lesions in the liver with Tc-99m-GSA SPECT--correlation with pathological evaluation and MRI findings.

To evaluate the effect of technetium-99m-labeled DTPA-galactosyl human serum albumin (Tc-99m-GSA) SPECT imaging for qualitative diagnosis of hepatic lesions. The subjects were 29 patients with pathologically confirmed hepatic lesions (21 malignant and 8 benign lesions). SPECT data were obtained at about 30 minutes after injecting 185 MBq (5 mCi) of Tc-99m-GSA. The GSA SPECT findings were compared with those of pathological evaluation and T2-weighted MR images (T2WI). Of 29 lesions, 17 showed decreased accumulation, and three exhibited increased accumulation. The other nine lesions were undetectable. The malignant lesions which showed increased accumulation were all well differentiated hepatocellular carcinomas (HCCs). One of the eight benign lesions exhibited increased accumulation. The three lesions which showed increased accumulation of GSA exhibited hypointensity on T2WI, whereas the malignant lesions which showed decreased accumulation of GSA exhibited hyperintensity on T2WI. The GSA SPECT findings correlate well with those of T2WI. GSA SPECT may be useful for qualitative diagnosis of focal liver lesions. If a lesion is suspected of being HCC, increased accumulation may indicate well differentiated HCC.

Adenocarcinoma↗

Angiotensin II stimulates both aldosterone secretion and DNA synthesis via type 1 but not type 2 receptors in bovine adrenocortical cells.

Angiotensin II (Ang II) type 2 receptor (AT2) has been shown to counteract the type 1 receptor (AT1)-mediated biological actions of Ang II in the cardiovascular system. The biological significance of AT2 receptor in the adrenals however remains unknown. In the present study, we investigated the roles of AT1 and AT2 receptor subtypes in the regulation of aldosterone secretion and DNA synthesis in bovine adrenocortical zona glomerulosa cells in vitro. Ang II (1 mumol/l)-stimulated aldosterone secretion was completely suppressed by AT1 antagonist CV-11974 but not affected by AT2 receptor antagonist PD-123319. Effects on DNA synthesis were investigated by determining the incorporation of BrdU into the nuclei of the cultured zona glomerulosa cells. Ang II (1 mumol/l)-stimulated DNA synthesis of the cells was also completely suppressed by CV-11974 but not by PD-123319. These results suggest that AT1 receptor but not AT2 receptor is the predominant receptor subtype which mediates the Ang II-stimulated aldosterone secretion and cell growth in bovine adrenocortical cells.

Adrenal Cortex↗

Differential expression of type IV collagen isoforms, alpha5(IV) and alpha6(IV) chains, in basement membranes surrounding smooth muscle cells.

Smooth muscle is composed of cigar-shaped, non-striated cells, each of which is encapsulated by a basement membrane and forms the contractile portion of tubular organs such as the gastrointestinal tract, pulmonary tract, genitourinary tract, and vasculature, in which slow and sustained contractions are needed. We examined basement membranes produced by smooth muscle cells and, using alpha(IV) chain-specific monoclonal antibodies, analyzed type IV collagens in these organs. Detailed distribution analysis of the alpha chains in normal and Alport cases by use of specific antibodies indicated that there are at least three molecular forms of type IV collagen, [alpha1(IV)]2alpha2(IV),alpha3(IV)alpha4(IV)alpha5+ ++(IV), and alpha5(IV)/alpha6(IV). Smooth muscle cells in the urinary bladder and uterus were enclosed by basement membranes composed of alpha1, alpha2, alpha5, and alpha6 chains. The same alpha chains were present around smooth muscle cells in the muscular layer of the fundus of the stomach, whereas those in the antrum and further distal side of the gastrointestinal tract expressed mostly alphal and alpha2 chains. In addition, immunostaining analysis of the vasculature also showed that most of the smooth muscle cells were positive for alpha1 and alpha2 chains; however, alpha5 and alpha6 chains were also expressed by smooth muscle cells in the aorta and some arteries where blood pressure changes significantly. These results suggest that the smooth muscle cells enclosed by alpha5/alpha6-containing basement membranes might have some particular function related to mechanical stress or tensile strength during the characteristic contractile activity of tubular organs.

Amino Acid Sequence↗

A surgical technique for a vertebral column autograft using the intervertebral disc for cervical disc disease.

We describe a surgical technique for a vertebral column autograft using the intervertebral disc for cervical disc disease for patients whose major problem is not spinal instability. Of a total of 41 patients with cervical disc disease suffering from cervical spondylotic radiculomyelopathy, 33 patients were operated on at one level and 8 patients were operated on at two levels. Postoperative X-ray film showed some movement at the "operated" disc level in all patients (average postoperative follow-up period was 43 months, range two years to 5 years). A significant decrease in motion in the extension position was observed postoperatively (p < 0.0001), but no significant difference was observed between the preoperative motion and the postoperative motion in the flexion position. Anterior angulation was found in two (5%) of the 41 patients. This surgical procedure has two major advantages: 1) no complications related to the iliac donor site, allowing early patient mobilization; 2) the extensive posterior spur can be removed safely and easily under a wide operative field. We believe that this surgical procedure is suitable for preserving the mobility of the spine and may avoid stress concentration at adjacent levels of the "operated" disc. However, in patients whose major problem is spinal instability, anterior cervical fusion should be performed.

Adult↗

Seroprevalence of immunoglobulin G antibodies against Helicobacter pylori among endoscopy personnel in Japan.

BACKGROUND: The seroprevalence of immunoglobulin G antibodies against Helicobacter pylori in endoscopy personnel was determined to investigate whether gastrointestinal endoscopists and endoscopy nurses in Japan are at an increased risk for H. pylori infection and to clarify risk factors for H. pylori infection during endoscopy. METHODS: One hundred twenty-one gastrointestinal endoscopists and endoscopy nurses provided personal information, including their observance of infection-control measures, by means of self-administered questionnaire. One hundred one age-matched healthy individuals undergoing routine physical examinations served as controls. Serum samples from each subject were examined with enzyme-linked immunosorbent assay for the presence of IgG antibodies against H. pylori. RESULTS: Among younger subjects (< 40 years old), endoscopists and endoscopy nurses had higher seropositive rates than did control subjects (p < 0.05). Among older subjects (> or = 40 years old), the seropositive rate did not differ between endoscopy personnel and control subjects (p = 0.2174). However, among older seropositive subjects, endoscopy personnel had significantly higher antibody titers than did control subjects (p < 0.01). Older seropositive endoscopists performed significantly more examinations per month than did their seronegative colleagues (p < 0.05). Furthermore, younger seropositive endoscopy nurses performed significantly more examinations per month than did seronegative nurses (p < 0.05). CONCLUSIONS: Gastrointestinal endoscopists and endoscopy nurses in Japan are at high risk for H. pylori infection. The risk of H. pylori infection is correlated with the frequency of endoscopic examinations, especially in older gastrointestinal endoscopists and younger endoscopy nurses.

Adult↗

Deferential roles of angiotensin receptor subtypes in adrenocortical function in mice.

The functional significance of angiotensin II (Ang II) receptor subtypes in adrenals remains unknown. Ang II receptor type 1a (AT1a) expression was localized by in situ hybridization to the zona glomerulosa and zona fasciculata, while AT1b was localized to the zona glomerulosa. Plasma aldosterone and corticosterone levels were measured after injection with Ang II or the type 2 receptor (AT2) agonist CGP-42112 in wild-type and AT1a deficient mice. Aldosterone and corticosterone levels were lower in AT1a deficient mice. Ang II increased plasma aldosterone levels in AT1a deficient mice, but to a lesser extent in mice pretreated with nonselective AT1a/AT1b antagonist, CV-11974. CGP-42112 did not affect aldosterone levels. Ang II increased corticosterone levels in wild-type mice but not in AT1a deficient mice. Results suggest Ang II stimulates aldosterone secretion via AT1a and AT1b in the zona glomerulosa and corticosterone secretion via AT1a in the zona fasciculata, and provide first evidence for differential roles of AT1a and AT1b in the adrenals.

Adrenal Cortex↗

Cerebrospinal fluid somatostatin in West syndrome: changes in response to combined treatment with high-dose pyridoxal phosphate and low-dose corticotropin.

Eighteen children with West syndrome (5-11 months of age) were selected to receive an oral dose of pyridoxal phosphate, (20-50 mg/kg) for 14 d. Seizures disappeared in one patient. The remaining 17 patients were treated with 0.01 mg/kg synthesized corticotropin intramuscularly for 2 weeks as an additional therapy. Seizures disappeared in all 17 patients within a few days after initiation of the corticotropin. Levels of somatostatin in the cerebrospinal fluid were as follows: 61.0+/-10.7 pg/ml before therapy, 34.2+/-6.4 pg/ml during pyridoxal phosphate therapy, and 26.8+/-4.2 pg/ml after 2 weeks corticotropin therapy. Somatostatin levels in untreated patients were higher (p < 0.05) than those of age-matched controls (35.7+/-11.8 pg/ml) and decreased (p < 0.05) after pyridoxal phosphate treatment. Somatostatin is a hypothalamic tetradecapeptide with excitatory effects on neurons and pyridoxal phosphate might subclinically influence neuronal excitation.

Adrenocorticotropic Hormone↗

Differential expression of transforming growth factor-beta and its receptors in hepatocytes and nonparenchymal cells of rat liver after CCl4 administration.

BACKGROUND/AIMS: Transforming growth factor-beta (TGF-beta) is a family of multifunctional proteins that regulate hepatocyte proliferation, and biosynthesis of the extracellular matrix. In this study we examined whether modulation of TGF-beta receptor expression contributes to the liver diseases. METHODS: The mRNA expression of TGF-beta1, TGF-beta type I receptor (TGFbetaRI), TGF-beta type II receptor (TGFbetaRII) and TGF-beta type III receptor (TGFbetaRIII) in rat livers injured by CCl4 administration was studied by Northern blotting. The mRNA expression patterns were confirmed by in situ hybridization. RESULT: The peak of TGF-beta1 mRNA expression was observed 48 h after acute intoxication with CCl4 in nonparenchymal cells. However, the levels of TGFbetaRI and TGFbetaRII mRNA expression decreased from 24 h to 48 h and from 12 h to 48 h, respectively, and returned to the normal level by 72 h. TGFbetaRII mRNA expression was depressed more and for longer than that of TGFbetaRI mRNA. Analysis in separated hepatocytes and nonparenchymal cells from the injured livers indicated that the mRNA changes occurred in hepatocytes. Nonparenchymal cells expressed TGFbetaRI and TGFbetaRII mRNAs at constant levels during liver regeneration. TGFbetaRIII mRNA, which also decreased after 12 h, was not apparent in hepatocytes but only in nonparenchymal cells. CONCLUSIONS: These observations suggest that: (i) whenever TGF-beta1 is increased in CCl4-treated livers, it may induce liver fibrogenesis via nonparenchymal cells; (ii) the mitoinhibitory effect of TGF-beta1 on hepatocytes is transiently relieved by down-regulation of TGF-beta receptors for 72 h post-damage; and (iii) the resistance to TGF-beta growth inhibition between 24 to 48 h may be predominantly due to down-regulation of the expression of TGFbetaRII.

Animals↗

Gene expression and roles of angiotensin II type 1 and type 2 receptors in human adrenals.

Although stimulation of aldosterone secretion is one of the functions of angiotensin II, the gene expression and biological significance of the angiotensin II receptor subtypes, AT1 and AT2, in the human adrenal have not been characterized. We therefore investigated the transcription levels of the receptor subtype genes and their roles in regulation of steroid secretion by human adrenals. The expression of AT1 and AT2 receptor mRNA was assessed by reverse transcription-polymerase chain reaction followed by Southern blot analysis in normal adrenocortical tissues (n = 6) and a series of adrenal tumour tissues: aldosterone-producing adrenocortical adenoma (n=6), Cushing's syndrome (n = 6) and pheochromocytoma (n = 6). The role of the two receptor subtypes in steroid secretion in vitro was examined by incubating the tissue with angiotensin II(1 microM) with or without the selective AT1 antagonist CV-11974 (1 microM). Both AT1 and AT2 receptor mRNA transcripts were demonstrated in all of the human adrenal tissues tested. Angiotensin II-induced aldosterone secretion was suppressed 50% upon the addition of CV-11974. The selective AT2 agonist CGP-42112 increased aldosterone secretion by 55% over the control, which was not suppressed by CV-11974. Angiotensin II and CGP-42112 did not affect cortisol secretion. These results suggest that both AT2 and AT1 receptors may be involved in the regulation of aldosterone secretion and tumorigenesis of the human adrenals.

Adenoma↗