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Biomedical subjects

K Mizuno

Publications and source records attributed to K Mizuno.

At least 73 records · Page 4Linked to original sources
PubMed2001

Overexpression of EXTL3/EXTR1 enhances NF-kappaB activity induced by TNF-alpha.

EXTL3/EXTR1 is a member of the EXT gene family, which may represent a class of glycosyltransferases involved in heparan sulfate biosynthesis. It is known that heparan sulfate interacts with a variety of proteins and is therefore implicated in various cellular responses. Here, we examined the effect of EXTL3 on nuclear factor-kappaB (NF-kappaB) activity stimulated by tumor necrosis factor-alpha (TNF-alpha), one of heparin-binding cytokine. The luciferase assay demonstrated that overexpression of EXTL3 enhanced TNF-alpha-induced NF-kappaB activity. This is confirmed with an electrophoretic mobility shift assay. However, EXTL3 did not affect the CD40-mediated NF-kappaB activation. The EXTL3 mutants lacking the amino terminus region failed to enhance the activity. The fluorescence of enhanced green fluorescent protein (EGFP)-fused EXTL3 was observed at the perinuclear region, whereas, the amino terminus-truncated mutant was found in a diffuse cytoplasmic region. These results suggest that EXTL3 may modulate NF-kappaB mediated by TNF-alpha.

CD40 Antigens↗
PubMed2001

The fibril structure of type V collagen triple-helical domain.

Although the triple-helical structure of fibrillar collagen is regarded in general as being quite similar, each type of collagen molecule has inherent characteristics in the triple-helical domain. Few studies have ever been performed in terms of the aggregate structure of the triple-helical domain of fibrillar collagen. Reconstituted aggregates from the purified triple-helical domain of each type of fibrillar collagen might amplify the subtle differences in the structural characteristics of each type of collagen molecule. In this study, the reconstituted aggregate structure of pepsin-treated type V collagen (type Vp collagen), that is, virtually its triple-helical domain was characterized by transmission electron microscopy. Pepsin-treated type I (type Ip) and type II (type IIp) collagen were compared with type Vp collagen. Unique features of the aggregate structure of the triple-helical domain of the type V collagen can be summarized as follows:These results suggested that the lateral packing of the triple-helical domain of type V collagen is determined by its molecular structure. The characteristics of type Vp collagen fibrils might be explained by their characteristic amino acid composition. A significant feature of the triple-helical domain of type V collagen is the high content of glycosylated hydroxylysine residues. Molecular model building of the collagenous structure suggests that a change in surface roughness is conspicuous by incorporating the glycosylated hydroxylysine residues. More than a ten-fold content of bulky glycosylated hydroxylysine residues in type V collagen compared to that of type I might have a significant influence on both the intermolecular and interfibrillar interactions of the triple-helical domain of type V collagen molecule.

Amino Acid Sequence↗
PubMed2001

Cell density-dependent death mode switch of cultured cortical neurons under serum-free starvation stress.

1. Cell death mode switch of cortical neurons from E17 rats was studied. Cells rapidly died under the serum-free condition. The time-course of cell death was markedly delayed by increasing cell density for primary culture in the trypan blue exclusion, LDH release, and MTT assays. 2. By analyzing cell death by the use of double staining using PI/TUNEL and PI/Annexin V combinations, the mode in the low density culture was found to be necrosis, while that in the high density culture was apoptosis. 3. The intracellular ATP level after the start of serum-free culture rapidly decline to 25% of 0-time level in the low density culture, but it was 60% in the high density culture. Both oligomycin and zVAD-fmk markedly decreased ATP levels and the population of TUNEL-positive neurons, while 3-aminobenzamide slightly increased these indices. 4. Thus. it is strongly suggested that the cell death mode switch from necrosis to apoptosis is closely related to intracellular ATP levels, and some conditioned medium factors observed in the high density culture may affect both ATP level and cell death mode switch.

Animals↗
PubMed2001

PI-3 kinase and IP3 are both necessary and sufficient to mediate NT3-induced synaptic potentiation.

Signaling mechanisms underlying neurotrophic regulation of synaptic transmission are not fully understood. Here we show that neurotrophin-3 (NT3)-induced potentiation of synaptic transmission at the neuromuscular synapses is blocked by inhibition of phosphoinositide-3 kinase, phospholipase C-gamma or the downstream IP3 receptors of phospholipase C-gamma, but not by inhibition of MAP kinase. However, neither stimulation of Ca2+ release from intracellular stores by photolysis of caged IP3, nor expression of a constitutively active phosphoinositide-3 kinase (PI3K*) in presynaptic motoneurons alone is sufficient to enhance transmission. Photo-uncaging of IP3 in neurons expressing PI3K* elicits a marked synaptic potentiation, mimicking the NT3 effect. These results reveal an involvement of PI3 kinase in transmitter release, and suggest that concomitant activation of PI3 kinase and IP3 receptors is both necessary and sufficient to mediate the NT3-induced synaptic potentiation.

Animals↗
PubMed2001

Phosphorylation of cofilin by LIM-kinase is necessary for semaphorin 3A-induced growth cone collapse.

Semaphorin 3A is a chemorepulsive axonal guidance molecule that depolymerizes the actin cytoskeleton and collapses growth cones of dorsal root ganglia neurons. Here we investigate the role of LIM-kinase 1, which phosphorylates an actin-depolymerizing protein, cofilin, in semaphorin 3A-induced growth cone collapse. Semaphorin 3A induced phosphorylation and dephosphorylation of cofilin at growth cones sequentially. A synthetic cell-permeable peptide containing a cofilin phosphorylation site inhibited LIM-kinase in vitro and in vivo, and essentially suppressed semaphorin 3A-induced growth cone collapse. A dominant-negative LIM kinase, which could not be activated by PAK or ROCK, suppressed the collapsing activity of semaphorin 3A. Phosphorylation of cofilin by LIM-kinase may be a critical signaling event in growth cone collapse by semaphorin 3A.

Actin Depolymerizing Factors↗
PubMed2001

Thrombin-induced p38 mitogen-activated protein kinase activation is mediated by epidermal growth factor receptor transactivation pathway.

Thrombin is a potent mitogen for vascular smooth muscle cells (VSMC) and has been implicated its pathogenic role in vascular remodelling. However, the signalling pathways by which thrombin mediates its mitogenic response are not fully understood. We have previously reported that thrombin activates p38 mitogen-activated protein kinase (p38 MAPK) by a tyrosine kinase-dependent mechanism, and that p38 MAPK has a role in thrombin-induced mitogenic response in rat VSMC. In the present study, we examine the involvement of epidermal growth factor (EGF) receptor in thrombin-induced p38 MAPK activation. We found that thrombin induced EGF receptor tyrosine phosphorylation (transactivation) in A10 cells, a clonal VSMC cell line. A selective inhibitor of EGF receptor kinase (AG1478) inhibited the p38 MAPK activation in a dose-dependent manner, whereas it had no effect on the response to platelet-derived growth factor (PDGF). EGF receptor phosphorylation induced by thrombin was inhibited by BAPTA-AM and GF109203X, which suggest a requirement for intracellular Ca(2+) increase and protein kinase C. We next examined the effect of AG1478 on thrombin-induced DNA synthesis. AG1478 inhibited thrombin-induced DNA synthesis in a dose-dependent manner. In contrast, PDGF-induced DNA synthesis was not affected by AG1478. In conclusion, these data suggest that the EGF receptor transactivation and subsequent p38 MAPK activation is required for thrombin-induced proliferation of VSMC.

Blotting, Western↗
PubMed2001

Neuroprotection by estrogen via extracellular signal-regulated kinase against quinolinic acid-induced cell death in the rat hippocampus.

Extracellular signal-regulated kinase (ERK) belongs to the family of mitogen-activated protein kinases (MAPKs), which are serine-threonine kinases activated by phosphorylation in response to a variety of mitogenic signals. We previously reported that 17 beta-estradiol rapidly activates ERK in the rat hippocampus. However, the physiological role of this rapid activation of ERK by estrogen in vivo has not yet been elucidated. This study investigated whether ERK may participate in mediating the neuroprotective effects of estrogen against quinolinic acid (QA) toxicity in the rat hippocampus in vivo. Injection of QA into the hippocampi of male rats produced a loss of Nissl-stained neurons in the CA1 after 24 h. Prior administration of 17 beta-estradiol (50 pmol/animal) to the ventricles prevented the QA-induced decrease in Nissl-stained neurons. Pretreatment with U0126, an inhibitor of MAPK/ERK kinase, inhibited the rapid activation of ERK by 17 beta-estradiol in the rat hippocampus. Moreover, the neuroprotective effects of 17beta-estradiol against QA toxicity were blocked by the pretreatment with U0126. U0126 alone did not produce a loss of neurons. These results indicate that ERK mediates estrogen neuroprotection after QA toxicity in the rat hippocampus.

Animals↗
PubMed2001

Heightened ability of monocytes from sarcoidosis patients to form multi-nucleated giant cells in vitro by supernatants of concanavalin A-stimulated mononuclear cells.

The main immunocompetent cells in sarcoidal lesions are epithelioid cells and multi-nucleated giant cells (MGC), both of which are derived from monocyte-macrophage lineage cells. To understand further the relevance of monocytes in sarcoidosis, we examined in vitro MGC formation using monocytes from sarcoidosis patients, patients with other granulomatous diseases (OGD) and healthy control subjects. The supernatant of concanavalin A-stimulated peripheral blood mononuclear cells (conditioned medium) generated Langhans type-MGC and foreign body type-MGC from monocytes. Conditioned medium from any three groups had the same ability to form MGC from normal monocytes. On the other hand, MGC were more highly formed using monocytes from sarcoidosis patients than from other groups. When macrophages induced by treatment of monocytes with macrophage colony-stimulating factor (M-CSF) were used, the rate of MGC formation in sarcoidosis patients was about threefold or fourfold as much as that in OGD patients or healthy controls, respectively. Oxidized ATP inhibited MGC formation in all groups. The susceptibility of monocytes cultured in conditioned medium for 24 h to 2'- and 3'-o-(4-benzoyl-benzoyl)ATP-mediated cytolysis was significantly higher in sarcoidosis patients than other groups. These findings suggest that the ability of monocytes to form MGC through P2x7 receptors is enhanced in sarcoidosis patients.

Adenosine Triphosphate↗
PubMed2001

Sociomedical aspects of epileptic patients: their employment and marital status.

We examined the employment and marital status of adult patients with epilepsy who did not have mental retardation and who had been treated at Hirosaki University Hospital, Hirosaki, Japan, for more than 5 years. The present study included 278 patients (142 males and 136 females) ranging from 20 to 60 years of age. We investigated the occupational status of the subjects and found that 168 had permanent jobs, but 41 patients were unemployed at the time of this survey. The proportion of the patients whose seizures were controlled at the time of this survey was 68% (114/168) in the group having permanent jobs, and 22% (9/41) in the unemployed group. Forty cases answered that they had resigned from their jobs due to occurrence of epileptic seizures. Of these patients, 13 were dismissed and 27 resigned voluntarily due to the potential for seizures. As to relationship between jobs and neuropsychiatric complications, the incidence of a past history of psychotic states in the unemployed group was significantly higher than that in the employed group. As to marital status, 13 males and 16 females (n = 29) had experienced divorce. Seven cases (two males and five females) had answered that epilepsy had been the reason for their divorce. We conclude that epilepsy or epileptic seizures have various negative effects on the patient's social life.

Adult↗
PubMed2001

Development of sucking behavior in infants who have not been fed for 2 months after birth.

BACKGROUND: Term infants are able to develop sucking behavior after birth. However, the requirements for this development have not yet been fully elucidated. In the present study, we investigated whether an oral-feed practice is necessary for the development of sucking behavior and whether non-nutritive sucking could support the development of this behavior while the infants cannot be fed. METHODS: Subjects of the present study were four term or near-term infants who had never been fed orally for approximately 2 months after birth because of gastrointestinal problems. Sucking pressure was measured with a silicone tube inserted into an artificial nipple and sucking efficiency was calculated during an entire feeding. RESULTS: Sucking pressure, frequency and duration at the first time oral feed corresponded to values obtained for normal-term infants at their first oral feed. Although infants who had not received oral feeding demonstrated poor sucking abilities initially, sucking performance improved with practice. Sucking parameters measured 1 month since oral feeding was initiated in these infants corresponded to values obtained for normal-term infants at 1 month of age. Although infants who had not received oral feeding had often sucked a pacifier for sedation, this non-nutritive sucking did not result in the development of sucking behavior. CONCLUSIONS: These results demonstrate that an oral feeding practice is necessary for the development of sucking behavior and that non-nutritive sucking does not affect the development of sucking.

Enteral Nutrition↗
PubMed2001

Primary and salvage urethroplasty using Mathieu meatal-based flip-flap technique for distal hypospadias.

BACKGROUND: Numerous surgical procedures have been attempted for correction of distal hypospadias. The Mathieu procedure was employed in this study for secondary cases as well as primary cases. METHODS: The length of the skin flap is determined by measuring the distance from the meatus to the glans tip and then the ventral meatal-based skin flap is incised. The proximal skin flap is flipped and anastomosed to the distal urethral plate. Additionally, subcutaneous tissue of the flipped flap is sutured to cover the original suture lines completely. RESULTS: The Mathieu urethroplasty was successful in one stage in 13 of the 16 primary repair cases (81%). Twelve of the 13 (92%) who underwent a secondary Mathieu procedure were successfully repaired with no problems. An overall success rate of 86% was achieved at the first operation for both primary and secondary cases. In the remaining 14% of the cases, success was achieved with only one additional procedure. CONCLUSIONS: The Mathieu flip-flap procedure is feasible for relatively short urethral defects if the ventral penile skin demonstrates adequate mobility and there is no chordee. Even in patients who require revisional hypospadias surgery, the Mathieu procedure can give excellent functional and cosmetic results.

Adolescent↗
PubMed2001

One-stage repair of moderately severe hypospadias using a transverse preputial tubularized island flap.

BACKGROUND: Transverse preputial tubularized island flap (TPTIF) urethroplasty has been used for the repair of moderately severe hypospadias since Duckett described the procedure in 1980. In spite of the excellent results reported by Duckett, subsequent studies showed high complication rates. A TPTIF procedure modified to reduce the complication rate is presented. METHODS: Between 1996 and 1997, 13 boys with moderately severe hypospadias were repaired with the TPTIF procedure. Patient age ranged from 10 months to 3 years with an average age of 23 months. To prevent urethrocutaneous fistula, the neourethra was constructed with a two-layer closure and the portion of anastomosis was wrapped between the native urethra and the neourethra with the tissue of the corpus spongiosum. RESULTS: The moderately severe hypospadias was repaired without complication in 12 of 13 patients. A urethrocutaneous fistula developed at the midshaft of the penis in one patient. No meatal stenosis, urethral stricture or diverticulum developed. CONCLUSION: Transverse preputial tubularized island flap urethroplasty provided excellent cosmetic and functional results for moderately severe hypospadias, and postoperative complications could be decreased by the two-layer closure of the neourethra and application of the wrapping technique of the proximal anastomosed portion with corpus spongiosum tissue.

Child, Preschool↗
PubMed2001

Tubularized incised-plate urethroplasty for secondary hypospadias surgery.

PURPOSE: Reoperation for failed hypospadias has been considered to be seriously bothersome because abundant penile skin does not tend to remain for urethroplasty or for penile shaft skin coverage. In this study, the tubularization of incised urethral plate was employed for those who had no excessive penile skin after failure of hypospadias repair. METHODS: Five patients with hypospadias underwent tubularized incised-plate urethroplasty as salvage surgery. The surgical techniques necessary for the performance of the reoperation were not different from those for the primary repair. The urethral plate was incised sufficiently deeply in its midline from the tip of the glans to the regressed meatus. The incised urethral plate was tubularized without tension over a catheter of an appropriate size. RESULTS: Four of those who underwent secondary tubularized incised-plate urethroplasty were successfully repaired without complications. A urethrocutaneous fistula occurred at the corona in the remaining patient. CONCLUSIONS: The absence of preputial skin in reoperative cases makes tubularized incised-plate urethroplasty the ideal option, although the series was small and postoperative duration is still short. In addition, this procedure can give excellent functional and cosmetic results even in patients who require revisional hypospadias surgery.

Adolescent↗
PubMed2001

The modified Koyanagi repair for severe proximal hypospadias.

OBJECTIVE: To report the method and results of a modified one-stage Koyanagi repair (urethroplasty with a parameatal-based and fully extended circumferential foreskin flap) to preserve the vascularity to the peripheral portion of the neourethra, in the repair of severe hypospadias. PATIENTS AND METHODS: Using a skin-incision line as in the original Koyanagi repair, a circumferential incision is made approximately 5 mm proximal to the corona and the urethral plate incised as for repair of chordee. A U-shaped skin incision is then made surrounding the meatus, extended to the dorsal prepuce for approximately 8 mm and parallel to the first incision. The tissue between the prepuce and dartos is dissected on the dorsal side to fix the prepuce as a neourethra to the dartos and to maintain blood supply. After mobilizing the loop-shaped skin flap through the button-hole of the pedicle, the internal and external sides of the loop are sutured to construct a neourethra. Twenty patients (aged 10 months to 9 years) with severe proximal hypospadias underwent the one-stage modified Koyanagi repair. RESULTS: The repair was successful after the initial procedure in 14 patients, but urethrocutaneous fistulae developed in three and meatal stenosis in three. The overall success rate was thus 70%. CONCLUSIONS: There were fewer complications than reported with the original Koyanagi repair, suggesting that the attempted vascular preservation of the neourethra was effective.

Child↗
PubMed2001

Histologic analysis of the implanted cartilage in an exact-fit osteochondral transplantation model.

PURPOSE: Osteochondral transplantation is one of the useful treatments for articular cartilage defect. However, the histologic change of the implanted cartilage has not been reported in detail. We investigated the histology of exact-fit osteochondral transplants used to repair articular cartilage defects in an animal model. TYPE OF STUDY: This was a nonrandomized control study using an animal model. METHODS: Sixteen skeletally mature female Japanese white rabbits were used in the study. The region of the femoral groove was selected as the site for the osteochondral defect. A full-thickness cylindrical defect (7 mm in diameter and 7 mm in depth) through the articular cartilage and into the subchondral bone was made using the Osteochondral Autograft Transfer System (Arthrex, Naples, FL). The entire osteochondral fragment was removed and then returned to its original site in the femoral condyle precisely. Thus, the defect was repaired with an autogenous osteochondral transplantation of exactly the same size and configuration as the defect. Specimens were obtained 2, 4, 12, and 24 weeks postoperatively and were analyzed both macroscopically and histologically. RESULTS: Macroscopically, there was smooth continuity of the articular surface and the integration of the graft to the normal host cartilage. However, histologic examination showed that the layer of the grafted cartilage was thicker than that of the normal host cartilage and the extracellular matrix of the implanted cartilage exhibited a stronger staining pattern with safranin-O fast green than the normal cartilage. Cell density was higher in the grafted cartilage, particularly in the middle and the deep zones. Round and polygonal hypertrophic clusters of chondrocytes were observed in the middle and deep zones of the grafted cartilage. CONCLUSIONS: The histologic properties of the exact-fit implanted cartilage were different from that of normal articular cartilage. Further investigation of mechanical and structural properties of grafted cartilage is necessary to verify the long-term effects of osteochondral transplantation.

Animals↗
PubMed2001

Hemangiomatosis presenting as meralgia paresthetica.

The authors present a 15-year-boy with meralgia paresthetica caused by the recurrence of a diffuse congenital hemangiomatosis in the pelvic region. Relief of the patient's symptoms was achieved by neurolysis of the lateral femoral cutaneous nerve in the thigh and partial excision of the tumor. To our knowledge, hemangiomatosis has never been suggested as a cause of meralgia paresthetica.

Adolescent↗