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Biomedical subjects

T Miyazaki

Publications and source records attributed to T Miyazaki.

At least 19 recordsLinked to original sources

New orally active serine protease inhibitors: structural requirements for their good oral activity.

Synthesis and structural requirements for good oral activity of a series of para-substituted benzoyl esters of 4-hydroxybenzamidine serine protease inhibitors are described. The structure required for good oral activity was found to be general formula II whose corresponding ester has to be hydrolyzed in the intestine before absorption through the mucous membranes or in plasma after absorption. Biological evaluation of oral absorption using plasma anti-trypsin activity was useful for rapid evaluation. By measuring their actual plasma concentrations after oral administration, compounds 14 and 16b were confirmed to show good area under the plasma concentration-time curves (AUC). Their plasma concentrations corresponded to their plasma anti-trypsin activity. Structure-oral activity relationships are discussed.

Administration, Oral

Critical role of the interleukin 2 (IL-2) receptor gamma-chain-associated Jak3 in the IL-2-induced c-fos and c-myc, but not bcl-2, gene induction.

The interleukin 2 receptor (IL-2R) consists of three subunits, the IL-2R alpha, IL-2R beta c, and IL-2R gamma c chains. Two Janus family protein tyrosine kinases (PTKs), Jak1 and Jak3, were shown to associate with IL-2R beta c and IL-2R gamma c, respectively, and their PTK activities are increased after IL-2 stimulation. A Jak3 mutant with truncation of the C-terminal PTK domain lacks its intrinsic kinase activity but can still associate with IL-2R gamma c. In a hematopoietic cell line, F7, that responds to either IL-2 or IL-3, overexpression of this Jak3 mutant results in selective inhibition of the IL-2-induced activation of Jak1/Jak3 PTKs and of cell proliferation. Of the three target nuclear protooncogenes of the IL-2 signaling, c-fos and c-myc genes, but not the bcl-2 gene, were found to be impaired. On the other hand, overexpression of the dominant negative form of the IL-2R gamma c chain, which lacks most of its cytoplasmic domain, in F7 cells resulted in the inhibition of all three protooncogenes. These results provide a further molecular basis for the critical role of Jak3 in IL-2 signaling and also suggest a Jak PTK-independent signaling pathway(s) for the bcl-2 gene induction by IL-2R.

Animals

IL-2 signaling involves recruitment and activation of multiple protein tyrosine kinases by the IL-2 receptor.

The IL-2 receptor (IL-2R) consists of three subunits, the IL-2R alpha, IL-2R beta, and IL-2R gamma chains, the last of which is also used in the receptors for IL-4, IL-7, IL-9, IL-13, and IL-15. The IL-2-induced proliferative signals emanate from the cytoplasmic domains of IL-2R beta and IL-2R gamma, but the nature and function of the signaling molecules that transmit these signals are not fully understood. Here we summarize our current understanding of the mechanisms by which IL-2R transmit signals by using multiple protein kinases. In fact, at least four protein tyrosine kinases (PTKs) are physically associated with IL-2R: p56lck (and its members), Syk PTK, and the Janus kinases, Jak1 and Jak3. cDNA expression studies revealed that the activation of these PTKs is critical for IL-2-induced proliferative signal transmission. Our findings indicate that a unique property of the IL-2R cytoplasmic domains is to recruit a variety of signaling molecules, which may suggest a mechanism by which these PTKs and other signaling molecules function in concert.

Animals

Activation of Stat5 by interleukin 2 requires a carboxyl-terminal region of the interleukin 2 receptor beta chain but is not essential for the proliferative signal transmission.

The high-affinity interleukin 2 (IL-2) receptor (IL-2R) consists of three subunits: the IL-2R alpha, IL-2R beta c, and IL-2R gamma c chains. Two members of the Janus kinase family, Jak1 and Jak3, are associated with IL-2R beta c and IL-2R gamma c, respectively, and they are activated upon IL-2 stimulation. The cytokine-mediated Jak kinase activation usually results in the activation of a family of latent transcription factors termed Stat (signal transducer and activator of transcription) proteins. Recently, the IL-2-induced Stat protein was purified from human lymphocytes and found to be the homologue of sheep Stat5/mammary gland factor. We demonstrate that the human Stat5 is activated by IL-2 and that Jak3 is required for the efficient activation. The cytoplasmic region of the IL-2R beta c chain required for activation of Stat5 is mapped within the carboxyl-terminal 147 amino acids. On the other hand, this region is not essential for IL-2-induced cell proliferation.

3T3 Cells

Three distinct IL-2 signaling pathways mediated by bcl-2, c-myc, and lck cooperate in hematopoietic cell proliferation.

Two interleukin-2 receptor-dependent signaling pathways have thus far been identified: the c-fos/c-jun induction pathway mediated by src family protein-tyrosine kinases and the c-myc induction pathway. Here, we provide evidence for the existence of a third, rapamycin-sensitive pathway, which results in the induction of another proto-oncogene, bcl-2. In the hematopoietic cell line BAF-B03, the expression of any two of lckF505 (an active form of p56lck), Bcl-2, or c-Myc is sufficient to promote transit of the cell cycle, regardless of the activation state of the third pathway. We also provide evidence that epidermal growth factor receptor signaling may act through the same pathway that involves p56lck. These studies demonstrate a novel approach to dissecting signaling pathways regulating cellular proliferation.

Animals

Epidural pressure measurements. Relationship between epidural pressure and posture in patients with lumbar spinal stenosis.

STUDY DESIGN: The relationship between epidural pressure and lumbar posture was assessed in patients with lumbar spinal stenosis. OBJECTIVES: This study was performed to assess the relationship between epidural pressure and lumbar posture in patients with lumbar spinal stenosis. METHODS: The study was performed on 10 patients who had cauda equina symptoms at the L4-L5 level. The catheter transducer was inserted into the epidural space through L5-S1 interlaminar space and placed at the L4-L5 disc level. This transducer was connected with an amplifier and a recorder. Epidural pressure was continuously measured in various postures. RESULTS: Local epidural pressure at the stenotic level was low in lying and sitting postures, and high in standing postures. Pressure was increased with extension, but decreased with flexion. The highest pressure was 116.7 +/- 38.4 mm Hg in standing with extension. CONCLUSION: Epidural pressure was significantly related to posture. These pressure changes correlated with the development of cauda equina symptoms. The increase of epidural pressure by posture may induce compression of the cauda equina. These pressure changes may explain the postural dependency in eliciting symptoms.

Aged

CNS myelinogenesis in vitro: time course and pattern of rat oligodendrocyte development.

Oligodendrocyte precursor cells that develop into myelin-forming cells of the central nervous system (CNS) were cultured from newborn rat brain to study how they proliferate and differentiate in normal conditioning medium, and their cell development was characterized by scanning electron microscopy (SEM) observation and immunocytochemical studies. We have identified A2B5-negative pre-O2A progenitor cells (so-called "type-1" oligodendrocytes) in the secondary cultures on the astrocyte feeder layer. These cells are very small (diameter: 3.5 microns), round, and glossy, and develop into the process-bearing O2A progenitor cells (called "type-2" oligodendrocytes), which also express myelin basic protein (MBP) both in the cell body and in their cell processes. Finally, they develop into mature oligodendrocytes (called "type-3" oligodendrocytes). After MBP expression is elicited in these cells and MBP accumulates in the cell process in the area in contact with the axon, these cells are capable of forming the myelin sheath. Therefore, we examined the mechanism of myelin-sheath formation of "type-3" oligodendrocytes using video time-lapse movies, and demonstrated that these cells initially sent out processes to search for axons several times before the onset of myelination. Then thick filopodia extended towards the axon, and at the same time, the axonal part of neuron moved forward. Finally the ruffling lamellipodial parts wrapped up the axon similarly to a transverse wave with the secured thick filopodial process on the axon acting as scaffolding. These results suggest that our experimental systems are useful in studying normal oligodendrocyte development and their cellular biochemistry, as well as investigating the mechanism of myelin formation by oligodendrocytes.

Animals

Glycine induces two distinct membrane currents in neonatal rat sympathetic preganglionic neurones in vitro.

1. The effects of glycine (Gly) on neonatal (12- to 16-day-old) rat sympathetic preganglionic neurones (SPNs) in transverse (500 microns) thoracolumbar spinal cord slices were studied by whole-cell patch-clamp techniques. 2. Gly elicited three types of membrane currents when applied to SPNs by pressure ejection (100 mM; 20-180 ms pulse duration): (1) an outward current (20/94 cells); (2) an inward current (30/94); and (3) a biphasic response (44/94) consisting of an outward followed by an inward current. 3. The Gly-induced outward current (IGly,(out)) had a mean reversal potential of -67 mV, was reversed in a low (5.7 mM) chloride Krebs solution, and was reversibly eliminated by strychnine (0.1-1 microM). 4. The Gly-induced inward current (IGly,(in)) had a mean reversal potential of -41 mV, was reduced in a Na(+)-free and increased in a high (15 mM) K+ solution; strychnine at the high concentration of 1 microM reduced the response by an average of 53%. 5. The electrophysiological and pharmacological characteristics of the biphasic response suggest that it was a combined response of outward and inward currents. 6. The results show that Gly elicits two distinct membrane currents, a Cl(-)-dependent outward current and a cationic inward current, which are mediated by strychnine-sensitive and strychnine-resistant Gly receptors. Functionally, activation of both types of Gly receptors reduces neuronal excitability and attenuates synaptic transmission.

Adrenergic Fibers

Serum pyridinolines as specific markers of bone resorption in hemodialyzed patients.

Serum levels of hydroxylysyl pyridinoline and lysyl pyridinoline were quantified in uremic patients undergoing maintenance hemodialysis and in healthy subjects. Pre-hemodialysis serum levels of hydroxylysyl pyridinoline and lysyl pyridinoline in the hemodialyzed patients were significantly higher than those in healthy subjects. Serum levels of hydroxylysyl pyridinoline and lysyl pyridinoline decreased significantly after hemodialysis with reduction rates of about 40%. Pre-hemodialysis serum levels of hydroxylysyl pyridinoline and lysyl pyridinoline correlated significantly with intact parathyroid hormone, osteocalcin and bone-specific alkaline phosphatase. Lysyl pyridinoline showed better correlations with these parameters than hydroxylysyl pyridinoline. Parathyroidectomy markedly decreased serum levels of hydroxylysyl pyridinoline and lysyl pyridinoline. These results indicate that serum pyridinolines, especially lysyl pyridinoline, may be used as specific biochemical markers of bone resorption in hemodialyzed patients.

Acid Phosphatase

C-type natriuretic peptide inhibits intimal thickening after vascular injury.

Recently we have found that C-type natriuretic peptide (CNP) inhibits proliferation of cultured rat vascular smooth muscle cells through an elevation of cGMP. We have now tested whether administration of CNP inhibits the development of intimal lesions induced by air-drying injury in rat common carotid arteries in vivo. CNP treatment (1 microgram/kg per min, iv infusion) for either 14 or 5 days resulted in 70% or 60% reduction, respectively, of intimal cross-section area 14 days after injury as compared with control rats. We also found that CNP potently stimulated cGMP production in injured carotid arteries with intimal thickening, but not in intact ones. These results indicate that GC-B, CNP specific receptor/guanylyl cyclase, is expressed at the sites of vascular injury, and that CNP might be efficacious in the prevention of restenosis caused by intimal thickening following coronary angioplasty.

Animals

Detection of (1-3)-beta-D-glucan in a rat model of aspergillosis.

The G test containing factor G, fractioned from the Limulus lysate, was used to detect (1-3)-beta-D-glucan in a rat model of aspergillosis. Aspergillus fumigatus strain MF-13, 1 x 10(4) conidia, were inoculated transtracheally into rats treated with cortisone acetate (100 mg/kg) and fed a low-protein (8%) diet. Increased serum (1-3)-beta-D-glucan was found on the sixth day after inoculation in concentrations of 370 +/- 178 pg/ml (mean +/- SD) in untreated controls, and 154 +/- 43 pg/ml in rats treated with 0.5 mg/kg of amphotericin B. On day 11 (1-3)-beta-D-glucan concentrations were 2,590 +/- 2,940 pg/ml and 448 +/- 442 pg/ml, respectively. The elevation in levels of (1-3)-beta-D-glucan increased in correlation with the elevation of galactomannan antigen titers; (1-3)-beta-D-glucan is thus measurable during experimental aspergillosis in rats.

Amphotericin B

Cadherin-11 expressed in association with mesenchymal morphogenesis in the head, somite, and limb bud of early mouse embryos.

Cadherin-11 (cad-11) is a novel member of the cadherin family of cell adhesion molecules, having recently been identified by means of the polymerase chain reaction. To study the function and expression of this molecule, we cloned mouse, cad-11 cDNA. Transfection of L cells with cDNA led them to acquire a typical cadherin-dependent cell-cell adhesiveness, and the L cells expressing cad-11 did not coaggregate with L cells expressing E-, P-, N-, or R-cadherin when they were mixed, indicating that this novel cadherin has a homophilic binding specificity, as found for other cadherins. To determine the developmental expression pattern of this molecule, we performed in situ hybridization analysis on early mouse embryos. Cad-11 first appeared in mesodermal layers only in the head and tail regions at the mid-to-late primitive streak stages. In the head, this appearance was followed by strong expression in mesenchymal tissues including branchial arches. In the trunk, the paraxial mesoderm initially did not express cad-11. However, as the somites formed, they expressed cad-11, and this expression was strictly correlated with their initial condensation and segregation from the presomitic mesoderm. The cad-11 expression in the somites was eventually restricted to sclerotome cells. As the limb buds developed, cad-11 appeared in the distal portion of the limb mesenchyme, and, at later stages, its expression was most evident at the peripheral mesenchyme. Cad-11 was thus expressed by restricted populations of mesenchymal cells in early embryos, although it was also expressed in parts of the neural tube, such as the optic vesicle and dorsal midline, and in part of the otic vesicle. As a step to investigate the role of cad-11 in mesenchymal cell adhesion, we dissociated the limb bud mesenchyme into single cells, pelleted them, and cultured them as aggregates. In these cultures, cad-11-positive cells clearly sorted out of the negative cell population, suggesting that cad-11 might be involved in selective association of mesenchymal cells. For comparison, we studied the expression of N-cadherin and found that the expressions of these two cadherins were differential, and complementary in some tissues. These results suggest that cad-11 is involved in specific associations of subsets of mesenchymal cells and also of some neural cells during early embryogenesis.

Amino Acid Sequence

Chronic cholecystitis following hemobilia due to traumatic intrahepatic injury.

We report a patient who presented with colicky abdominal pain, hematemesis, and melena following a blunt abdominal injury sustained in an automobile accident. Serologic tests suggested liver dysfunction and computed tomography (CT) revealed dilatation of the intrahepatic bile duct. Duodenoscopy demonstrated hemobilia originating from the duodenal papilla of Vater. Angiography revealed a pseudoaneurysm of the hepatic artery and angiographic embolization sucessfully stopped the hemorrhaging. However, even following the angiographic embolization, the patient remained symptomatic and repeat CT demonstrated thickening of the gallbladder wall and a lesion resembling a blood clot. We strongly suspected cholecystitis and performed a laparoscopic cholecystectomy. Grossly, the resected specimen revealed wall thickening with a congested mucosa and three blood clots; histologically, these changes were consistent with a diagnosis of chronic cholecystitis. The clinical message of this paper is that patients should be observed for this usual complication of hemobilia after liver injury.

Adolescent

Schmorl's nodes and low-back pain. Analysis of magnetic resonance imaging findings in symptomatic and asymptomatic individuals.

Magnetic resonance imaging (MRI) findings in cases with symptomatic and asymptomatic Schmorl's nodes have been analysed. In all symptomatic cases, the vertebral body marrow surrounding the Schmorl's node was seen as low signal intensity on T1-weighted images and as high signal intensity on T2-weighted images. It was confirmed by histological examination that the MRI findings indicated the presence of inflammation and oedema in the vertebral bone marrow. These MRI findings were not seen in asymptomatic individuals. Inflammatory changes in the vertebral body marrow induced by intraosseous fracture and biological reactions to intraspongious disc materials might cause pain. We postulate that after fracture healing and subsidence of inflammation, the Schmorl's nodes become asymptomatic, in analogy with old vertebral compression fractures. MRI is not only useful in detecting the recently developed Schmorl's nodes but also in differentiating between symptomatic and asymptomatic Schmorl's nodes.

Adolescent

Fluctuation of blood pressure and pulse rate during colostomy irrigation.

PURPOSE: The aim of this study was to determine the effects of colostomy irrigation on the vital signs of patients with left colostomy. METHODS: Twenty-two consecutive patients who underwent abdominoperineal resection for cancer of the lower rectum and had left lower quadrant end colostomy were included in this study. Subjective symptoms, blood pressure, and pulse rate during the first irrigation were investigated. RESULTS: Fluctuation of blood pressure during instillation was 8.0/8.5 mmHg (average) and 25.0/17.9 mmHg during evacuation. Fluctuation of pulse rate was 5.5 per minute (average) during instillation and 11.5 per minute during evacuation. The number of subjects who showed more than 20% fluctuation of systolic pressure was 12 (54.5 percent) and that of diastolic pressure was 14 (63.6 percent). One of 22 patients complained of illness during irrigation. CONCLUSION: Although colostomy irrigation showed no significant effects on vital signs in the majority of patients, it caused a significant reduction in both blood pressure and pulse rate in a small number of patients. Careful attention should be paid to vital signs considering the possibility of such effects, especially on the initial irrigation.

Aged

Formation of 6 beta-OH-deoxycorticosterone from deoxycorticosterone by A6 cells.

We have investigated the metabolism of deoxycorticosterone in A6 cells, a continuous cell line derived from the kidney of Xenopus laevis. A6 cells at confluence were incubated with serum-free culture media containing 2.5 microM [3H] deoxycorticosterone. When radioactive compounds in incubation media were separated by reverse-phase high-pressure liquid chromatography, the formation of polar metabolites was observed. One component of polar metabolites cochromatographed with 6 beta-hydroxydeoxycorticosterone. In order to identify this component more rigorously, large scale cultures were performed and this compound was separated and purified by reverse-phase high-pressure liquid chromatography. The purified material was analyzed by gas chromatography-mass spectrometry, secondary ion mass spectrometry, and nuclear magnetic resonance spectroscopy. These procedures revealed that this material was 6 beta-hydroxydeoxycorticosterone.

Animals