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Biomedical subjects

T Sugimoto

Publications and source records attributed to T Sugimoto.

At least 559 records · Page 31Linked to original sources

Distribution of primary afferent fibers projecting from hindlimb cutaneous nerves to the medulla oblongata in the cat and rat.

The dorsal column nuclear complex, one of the most important relays for tactile perception, has well been known to be somatotopically organized. Topographical arrangements of terminal sites of individual cutaneous nerves within the dorsal column nuclei, however, have not been examined systematically, although many studies have been done upon primary afferents to the medulla oblongata, including the dorsal column nuclear complex. Thus, in the present study, distribution of primary afferent fibers projecting from the hindlimb cutaneous nerves to the medulla oblongata was examined in the cat and rat by means of the transganglionic transport method with horseradish peroxidase. Cutaneous primary afferent fibers projecting from the hindlimb to the medulla oblongata were distributed mainly in the ipsilateral gracile nucleus. Terminal labeling in the gracile nucleus was seen at all rostrocaudal levels of the nucleus, occasionally including the nuclear part straddling the midline (the median or accessory nucleus). The labeled axon terminals in the gracile nucleus were more densely distributed in the middle and caudal parts of the nucleus than in the rostral part. Although the fields of termination of the hindlimb cutaneous nerves overlapped in the gracile nucleus, the foci of the terminal labeling of the nerves innervating the distal parts of the hindlimb were located more medially or dorsomedially than those of the nerves innervating the proximal parts. Terminal labeling was further found in a small zone immediately medial to the rostromedial border of the external cuneate nucleus. This hitherto undescribed zone (U zone) contained a small cluster of medium-sized neurons in the cat. Although no particular cell cluster was found in the U zone of the rat, convergence of the primary afferent fibers of the cutaneous nerve from the hindlimb appeared to occur as in the U zone of the cat.

Animals↗

Direct involvement of cAMP-dependent protein kinase in the regulation of alkaline phosphatase activity by parathyroid hormone (PTH) and PTH-related peptide in osteoblastic UMR-106 cells.

The present study was performed to characterize the participation of parathyroid hormone (PTH)- and PTH-related peptide (PTHrP)-responsive dual signal transduction systems [cAMP-dependent protein kinase (PKA) and Calcium/protein kinase C (Ca/PKC)] in the regulation of alkaline phosphatase (ALP) activity in osteoblastic osteosarcoma cells (UMR-106). Both human (h) PTH-(1-34) and hPTHrP-(1-34) at 10(-8) M stimulated ALP activity to the similar degree. Dibutyryl, cAMP (dbcAMP) (10(-5), 10(-4) M) and Sp-diastereoisomer of adenosine cyclic 3',5'-phosphorothioate (Sp-cAMPS), a direct stimulator of PKA (10(-4) M) also stimulated its activity. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, (10(-7), 10(-6) M) did not affect its activity, while calcium ionophores, A23187 and ionomycin (10(-7), 10(-6) M) inhibited it. Although Rp-diastereoisomer of adenosine cyclic 3',5'-phosphorothioate (Rp-cAMPS), a direct inhibitor of PKA, (10(-4) M) did not affect ALP activity by itself, it significantly antagonized not only Sp-cAMPS-induced increase in ALP activity, but also PTH- and PTHrP-induced one. The present study first indicated that the activation of PKA was directly involved and acted as a main pathway in the regulation of ALP activity by PTH and PTHrP in osteoblasts.

Alkaline Phosphatase↗

Molecular cloning and expression of a cDNA of the bovine prostaglandin F2 alpha receptor.

Capitalizing on the significant sequence homology comprising the transmembrane motif regions of known prostanoid receptor family, we targeted the cloning of a cDNA clone for prostaglandin (PG) F2 alpha receptor from a bovine corpus luteum cDNA library. By using several pairs of degenerated primers created from a common motif of transmembrane domains, polymerase chain reaction gave a clone SN463 carrying the homologous sequence, which covered transmembrane motif IV-VI of the thromboxane (TX) A2 receptor. This polymerase chain reaction product was used as a DNA probe for the following cross-hybridization, and a clone BC2211 carrying a 2.2-kilobase pair DNA insert was isolated. This clone encodes a protein of 362 amino acid residues (M(r) = 40,983) with seven potential transmembrane domains and represented significant overall sequence homology to human TXA2 receptor protein (34% in amino acid). Injection of the mRNA synthesized in vitro from the cloned cDNA into a Xenopus oocyte elicited electrophysiological response to PGF2 alpha. Ligand binding displacement in membranes of mammalian COS-7 cells transfected with the cDNA indicated the rank order of affinity of the receptor to PGs: PGF2 alpha > PGD2 > PGE2 > STA2, a TXA2 agonist. PGF2 alpha activated inositol phosphate formation in COS-7 cells transfected with receptor cDNA. Northern blot analysis and in situ hybridization indicated that the PGF2 alpha receptor mRNA is highly expressed and accumulated in corpus luteum. This is the first report on a successful cloning of functional receptor cDNA for PGF2 alpha.

Amino Acid Sequence↗

A subpopulation of olivocerebellar projection neurons express neuropeptide Y.

By neuropeptide Y (NPY) immunohistochemistry and in situ hybridization histochemistry with digoxigenin-labeled oligonucleotide probes for this peptide, a subpopulation of neurons in the caudal portions of the rat dorsal and medial accessory olives were found to express NPY immunoreactivity and mRNA. In the cerebellum, NPY-immunolabeled climbing fibers were distributed to the flocculus and parts of vermal cortex. The results suggest particular association of NPY with the climbing fiber system presumably mediating spinal, visual and vestibular inputs.

Animals↗

Cross-talk of parathyroid hormone-responsive dual signal transduction systems in osteoblastic osteosarcoma cells: its role in PTH-induced homologous desensitization of intracellular calcium response.

The present study was designed to characterize the cross-talk of parathyroid hormone (PTH)-responsive dual signal transduction systems (cAMP-dependent protein kinase (PKA) and calcium/protein kinase C [PKC]) and its participation in PTH-induced homologous desensitization of intracellular calcium ([Ca2+]i) in osteoblastic UMR-106 cells. Although our recent study revealed that prolonged (more than 2 h) pretreatment with PKC-activating phorbol ester, phorbol 12-myristate 13-acetate (PMA) significantly decreased the PTH-stimulated cAMP production, pretreatment with PMA (10(-7) and 10(-6) M) but not 10(-6) M 4 alpha-phorbol 12,13-didecanoate (PDD), incapable of activating PKC for 30 min significantly augmented 10(-7) M hPTH-(1-34)-stimulated cAMP production. H-7 (50 microM), a PKC inhibitor, significantly antagonized this PMA-induced effect. Pretreatment with 10(-6) M PMA for 30 min did not affect PTH receptor binding but significantly augmented a cAMP responsiveness to 10(-5) M forskolin and 1 microgram/ml cholera toxin. Pertussis toxin (0.5 microgram/ml) did not affect the PMA-induced augmentation of the PTH-stimulated cAMP production. PTH caused a complete homologous desensitization of [Ca2+]i response within 30 min. Pretreatment with 10(-4) M dibutyryl cAMP for 30 min and 6 h significantly reduced and completely blocked the PTH-induced increase in [Ca2+]i, respectively. Pretreatment with 10(-4) M Sp-cAMPs, a direct PKA activator, for 30 min completely blocked the PTH-induced increase in [Ca2+]i. Rp-cAMPS (10(-4) M), an antagonist of PKA, slightly but significantly antagonized the PTH-induced homologous desensitization of [Ca2+]i response. The present study indicates that the time of exposure to PKC activation is a critical determinant in modulating the cAMP system, while PKA activation counterregulatorily acts on the [Ca2+]i system, and that PKA activation is linked to the PTH-induced homologous desensitization of [Ca2+]i response.

Adenylyl Cyclases↗

Reactive oxygen metabolite-induced toxicity to cultured bovine endothelial cells: status of cellular iron in mediating injury.

We aimed to determine the status of iron in mediating oxidant-induced damage to cultured bovine aortic endothelial cells. Chromium-51-labeled cells were exposed to reaction mixtures of xanthine oxidase/hypoxanthine and glucose oxidase/glucose; these produce superoxide and hydrogen peroxide, or hydrogen peroxide, respectively. Xanthine oxidase caused a dose dependent increase of 51Cr release. Damage was prevented by allopurinol, oxypurinol, and extracellular catalase, but not by superoxide dismutase. Prevention of xanthine oxidase-induced damage by catalase was blocked by an inhibitor of catalase, aminotriazole. Glucose oxidase also caused a dose-dependent increase of 51Ci release. Glucose oxidase-induced injury, which was catalase-inhibitable, was not prevented by extracellular superoxide dismutase. Both addition of and pretreatment with deferoxamine (a chelator of Fe3+) prevented glucose oxidase-induced injury. The presence of phenanthroline (a chelator of divalent Fe2+) prevented glucose oxidase-induced 51Cr release, whereas pretreatment with the agent did not. Apotransferrin (a membrane impermeable iron binding protein) failed to influence damage. Neither deferoxamine nor phenanthroline influenced cellular antioxidant defenses, or inhibited lysis by non-oxidant toxic agents. Treatment with allopurinol and oxypurinol, which inhibited cellular xanthine oxidase, failed to prevent glucose oxidase injury. We conclude that (1) among the oxygen species extracellularly generated by xanthine oxidase/hypoxanthine, hydrogen peroxide induces damage via a reaction on cellular iron; (2) deferoxamine and phenanthroline protect cells by chelating Fe3+ and Fe2+, respectively; and (3) reduction of cellular stored iron (Fe3+) to Fe2+ may be prerequisite for mediation of oxidant-induced injury, but this occurs independently of extracellular superoxide or cellular xanthine oxidase-derived superoxide.

Allopurinol↗

Second messenger signaling of c-fos gene induction by parathyroid hormone (PTH) and PTH-related peptide in osteoblastic osteosarcoma cells: its role in osteoblast proliferation and osteoclast-like cell formation.

The present study was performed to clarify second messenger signaling in parathyroid hormone (PTH)-induced c-fos gene expression, to characterize the participation of the c-fos gene in the regulation of osteoblast proliferation and function as well as osteoclast-like cell formation by PTH and to compare these effects of PTH with those of PTH-related peptide (PTHrP). Both human (h) PTH-(1-34) and hPTHrP-(1-34) at 10(-8) M induced a transient c-fos gene expression to a similar degree in osteoblastic osteosarcoma cells, UMR-106. N6,O2'-dibutyryl adenosine 3',5'-cyclic monophosphate (dbcAMP) as well as Sp-diastereoisomer of adenosine cyclic 3',5'-phosphorothioate (Sp-cAMPS), an activator of cAMP-dependent protein kinase (PKA), induced a weak c-fos gene expression. Although Rp-diastereoisomer of adenosine cyclic 3',5'-phosphorothioate (Rp-cAMPS), an inhibitor of PKA, almost completely antagonized dbCAMP- and Sp-cAMPS-induced expression of c-fos gene, it did not cause an obvious inhibition of PTH- or PTHrP-induced expression. Phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C (PKC), induced an intense expression of the c-fos gene, while 4 alpha-phorbol 12,13-didecanoate (4 alpha PDD), incapable of activating PKC, and calcium ionophores (A23187 and ionomycin) did not. Protein kinase C inhibitor (H-7, 50 microM) completely blocked the expression of the c-fos gene by PTH as well as by PTHrP). Antisense oligodeoxynucleotides (as-ODN) complementary to c-fos mRNA, which have been shown to inhibit its mRNA translation, at 1 microM significantly antagonized PTH- and PTHrP-induced inhibition of [3H] thymidine incorporation and stimulation of osteoclast-like cell formation in the presence of osteoblasts, but not an increase in alkaline phosphatase activity, compared to control oligodeoxynucleotides with same nucleotides as as-ODN but with a random sequence. The present study indicates the involvement of PKC system in c-fos gene expression by PTH as well as PTHrP and also indicates the involvement of the c-fos gene in the regulation of bone cell physiology by PTH and PTHrP.

Cell Division↗

c-fos induction in the subnucleus caudalis following noxious mechanical stimulation of the oral mucous membrane.

c-fos expression in response to noxious mechanical stimulation of the peripheral receptive field was used to study the somatotopic representation of the oral mucous membrane and trigeminal cutaneous receptive fields in the trigeminal subnucleus caudalis and the cervical spinal dorsal horn. Fos-neurons (neurons with c-fos protein-like immunoreactivity) following stimulation of the cutaneous receptive field were found in laminae I and II of the first and second cervical segments of the spinal cord ipsilateral to the stimulation and exhibited a clear somatotopic segregation; i.e., the fos-neurons responding to receptive fields in the mandibular, maxillary, and ophthalmic divisions were arranged in a mediolateral sequence. On the other hand, fos-neurons responding to the oral mucous membrane were found in the similar laminae of the rostral pole of subnucleus caudalis bilaterally. They were all found in the dorsalmost part of the subnucleus without segregation; e.g., topographic distributions of fos-neurons resulting from the mandibular and maxillary gingival stimulations were indistinguishable. The central mechanism of trigeminal nociception is discussed with respect to the difference between intra- and extraoral receptive fields.

Animals↗

Translocation of protein kinase C alpha and zeta in rat glomerular mesangial cells cultured under high glucose conditions.

The activities and expression of protein kinase C isoenzymes were examined in glomerular mesangial cells cultured under high glucose conditions. Exposure of cells to high glucose concentrations (27.8 mmol/l) for more than 3 days resulted in a significant elevation of protein kinase C activities in the membrane fraction. Of the protein kinase C isoenzymes, the levels of protein kinase C alpha significantly increased in the membrane fraction after 3 days of exposure to glucose, and protein kinase C zeta increased after 5 days of exposure. Levels of protein kinase C delta and epsilon remained unchanged and protein kinase, C beta and gamma were not detected. These results indicate that protein kinase C alpha and zeta are translocated under high glucose conditions possibly through different mechanisms.

Animals↗

Meningiomas: correlation between MRI characteristics and operative findings including consistency.

The findings on magnetic resonance imaging (MRI) in 73 surgically verified intracranial meningiomas were correlated with their histology and consistency during resection. T 1-weighted imaging was least useful since most of the tumours were iso-intense, similar to cortical grey matter regardless of histology or tumour consistency. The signal intensity on T2-weighted images was found to best correlate with both the histology and consistency of the meningioma. Generally, the low intensity portion of the tumour on T2-weighted images indicated a more fibrous and harder character, while the higher intensity portions indicated a more soft character. Most of the fibroblastic meningiomas showed the features of a hard tumour while angioblastic tumours showed the features of soft tumours. Tumours predicted to be harder on MR imaging generally took longer to resect than softer ones, and this relationship was shown best for the larger tumours. Using linear regression analysis, it appears that operative time for soft tumours is more affected by factors other than tumour consistency. Blood loss during surgery was also unrelated to the consistency of the tumour. These results suggest that the histology and consistency of meningiomas may be predictable from findings on T2-weighted imaging, and this may also predict the difficulty and time required for resection.

Adolescent↗

Femoral and spinal bone mineral density in Japanese osteoporotics with hip fracture.

In the present study, bone mineral density (BMD) of femoral neck and lumbar spine was compared between 38 Japanese female patients with hip fracture (age 63-89 years, mean +/- SD 76 +/- 7 years) and 162 age-matched female controls (age 62-90 years, mean +/- SD 75 +/- 7 years). BMD was measured in the femoral neck and lumbar spine (L2-4) using dual-photon absorptiometry (Norland model 2600). BMD values of femoral neck as well as lumbar spine were significantly lower in patients with hip fracture than in controls (0.504 +/- 0.097 v 0.597 +/- 0.101, p < 0.01, for femoral neck; 0.661 +/- 0.146 v 0.720 +/- 0.128, p < 0.05, for lumbar spine). Patients with hip fracture and controls were stratified according to their BMD levels at two measuring sites, and the ratio of the number of patients and controls at each BMD level was calculated as an indicator of fracture rate. This ratio showed an exponential increase as the femoral neck BMD declined, but only a gradual increase as the lumbar spine BMD declined. Specificity-sensitivity analysis revealed that BMD values of 0.59 and 0.54 g/cm2 at the femoral neck provided a specificity of 52% and 68% with a sensitivity of 90% and 75%, respectively. These findings suggest that Japanese patients with hip fracture are more osteoporotic than age-matched controls and that the selective measurement of femoral neck would be useful for predicting the risk of hip fracture.

Aged↗

The problems of surgical treatment for cardiac myxoma and associated lesions.

Twenty-four patients with cardiac myxomas consisting of 22 left and 2 right atrial myxomas were operated on. All myxomas were removed with an excision of the attachment walls using a cardiopulmonary bypass. Two myxomas required a partial cardiopulmonary bypass from the femoral vein to the artery prior to operation because they were on the verge of becoming stuck in the atrioventricular valves and potentially causing shock. For embolic complications of myxoma, the embolus of the external carotid artery was extirpated before undergoing cardiac surgery. In a patient with pulmonary infarction, the infarcted lung was resected simultaneously. Another patient with a cerebral infarction received a clipping of an aneurysm which later appeared in the infarcted area. For associated cardiac lesions, two patients underwent a coronary artery bypass graft and one mitral valve replacement with tricuspid annuloplasty. In the former two cases, the myxoma was removed prior to coronary artery bypass grafting because the use of retrograde coronary perfusion was considered to be sufficient to protect the heart. In the latter case, the removal of the myxoma first disclosed a significant mitral lesion which had been masked by the huge myxoma. All patients but one, who died of pneumonia, showed a good recovery. In this series, the problems of surgical treatment for cardiac myxoma and associated lesions are also discussed.

Adolescent↗

The efficacy of non-clamping selective cerebral perfusion in distal aortic arch aneurysm repair: report of a case.

Successful repair of a distal aortic arch aneurysm without aortic cross-clamping was carried out in a 74-year-old man, using a combination of special separate cerebral perfusion and retrograde coronary perfusion, termed non-clamping selective cerebral perfusion. We believe that satisfactory results following aortic arch surgery in elderly patients can only be achieved through the prevention of emboli derived from an aortic cross-clamping site, and shortened ischemic time of the vital organs.

Aged↗

Pseudomyxoma peritonei due to adenocarcinoma of the lung: case report.

A rare case of pseudomyxoma peritonei whose primary site was presumed to be the lung is reported. A 76-year-old woman was admitted to Hospital presenting with progressive abdominal distention. She had been admitted twice, 2 and 1 year previously for the evaluation of high plasma carcinoembryonic antigen (CEA) level, of 11.6 ng/ml. Chest computed tomography (CT) scan and chest X-ray film on the third admission revealed a nodular lesion in the left lower lung field, and transbronchial lung biopsy (TBLB) revealed mucus-producing tall columnar epithelial carcinoma. Paracentesis revealed gelatinous ascitic fluid. At laparotomy, appendix and ovary were normal, and there were many small cystic tumors on the peritoneal surface and omentum. The patient died 2 years later, after repeated episodes of dynamic ileus. The lung and abdominal tumors gradually increased in size during the 2-year period, but she developed no respiratory symptoms. Based on both the clinical and pathophysiological findings, the final diagnosis made was pseudomyxoma peritonei whose origin was a lung adenocarcinoma.

Adenocarcinoma↗

Slow abnormal conduction in the low right atrium: its anatomic basis and relevance to atrial reentry.

To characterize slow abnormal conduction in the low right atrium, which is known to be responsible for atrial flutter, electrophysiologic findings were correlated with anatomic features in a canine model of atrial flutter with ligation of the crista terminalis in the midright atrium. Activation in the low right atrium was mapped with a patch electrode containing 52 bipolar electrodes and a multiplexing system. A particular region in the low right atrium showed atrioventricular node-like electrophysiologic properties, a rate-dependent conduction delay, and Wenckebach periodicity. This area coincided with an area responsible for slow conduction during atrial flutter and unidirectional block at its initiation. Both pilsicainide and E-4031 preferentially blocked conduction in the specific area, leading to the termination of atrial flutter. Although refractoriness could not explain the abnormal conduction, anatomic studies consistently found the specific region to be in or around a thick muscle bundle, that is, the crista terminalis, or a thick pectinate muscle branching from the crista, located perpendicular to the wavefront of the pacing impulse and atrial flutter. These electrophysiologic and anatomic findings suggest that slow abnormal and atrioventricular node-like conduction over a thick muscle bundle, which is a normal anatomic feature of the low right atrium, plays a role in the initiation, maintenance, and termination of atrial reentry.

Animals↗

Parvalbumin-like immunoreactivity in nerve fibres within human molar tooth pulps.

The root pulp contained two types of parvalbumin-immunoreactive nerve fibres, smooth and varicose. The smooth were relatively thick (mostly 3-5 microns in diameter) and sometimes had large swellings, whereas the varicose consisted of oval varicosities of 1-3 microns in long diameter with very fine strands interconnecting them. Most of the parvalbumin-immunoreactive root fibres were smooth, and the varicose were rare. Upon entering the pulp chamber, both types of fibre splayed out and ran towards the roof of the chamber and the pulp horn. In the subodontoblastic layer of the roof and horn, parvalbumin-immunoreactive fibres showed relatively straight chains of granules; they sent their granular terminals to the odontoblastic layer. Some parvalbumin-immunoreactive granules were observed in the predentine and some even in the dentine. The diameter of axons in the root pulp and the spatial distribution of terminals strongly suggest that the parvalbumin-immunoreactive fibres are mostly myelinated and sensory.

Dental Pulp↗

Protection of cultured rat gastric cells against oxidant-induced damage by exogenous glutathione.

BACKGROUND/AIMS: Reduced glutathione (GSH) is an intracellular protectant against oxidants. The present study determined whether extracellular GSH protects against oxidant damage or whether an uptake system of GSH is present in cultured gastric cells. METHODS: Hydrogen peroxide was generated by glucose oxidase and glucose. Cytotoxicity was assessed by 51Cr release. Intracellular GSH was assayed by the method of Tietze. RESULTS: Pretreatment with extracellular GSH decreased H2O2-induced 51Cr release. Treatment with GSH enhanced cellular GSH content. Protection by pretreatment with GSH was prevented by buthionine sulfoximine (an inhibitor of gamma-glutamylcysteine synthetase). Enhancement of intracellular GSH was also prevented by buthionine sulfoximine. Acivicin (an inhibitor of gamma-glutamyl transpeptidase) prevented intracellular accumulation of GSH from extracellular GSH. Cysteine was effective in preventing damage and enhancing intracellular GSH content, whereas both glutamine and glycine were not. CONCLUSIONS: Extracellular GSH protects cultured gastric cells from H2O2 damage by accelerating intracellular GSH synthesis; this is mediated by membrane-bound gamma-glutamyl transpeptidase acting on extracellular GSH (which supplies these cells with cysteine) and then by intracellular gamma-glutamylcysteine synthetase.

Animals↗

Meprin is predominantly involved in parathyroid hormone degradation by the microvillar membranes of rat kidney.

Hydrolysis of fragments of the C-terminal and mid-portions of parathyroid hormone (PTH) by a phosphoramidon-insensitive metallo-endopeptidase, previously purified by us from the microvillar membranes of rat kidney, and by the microvillar membranes of rat kidney themselves were investigated using a reverse-phase HPLC, and the amino acid sequences of each produced PTH metabolite were compared after their determination with an automated gas-phase protein sequencer. The results showed that both the purified microvillar endopeptidase and the microvillar membranes of rat kidney limited hydrolyzed human (h) PTH-(39-84) and hPTH-(39-68) mainly at peptide bonds flanked by a hydrophilic amino acid residue, where are characteristic for the purified microvillar endopeptidase but not for other known endopeptidase including endopeptidase 24.11. In addition, most of PTH metabolites generated by the microvillar membranes were insensitive to phosphoramidon and had amino acid sequences identical to those generated by the purified microvillar endopeptidase, indicating that these metabolites were produced by the enzyme integrated in the membranes. Since analysis of the N-terminal amino acid sequence of the purified microvillar endopeptidase revealed that it was structurally identical to rat meprin (EC 3.4.24.18), these results indicate that the purified microvillar endopeptidase or meprin is predominantly involved in PTH degradation by the microvillar membranes of rat kidney as an integral membrane protein.

Amino Acid Sequence↗