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Biomedical subjects

S Torii

Publications and source records attributed to S Torii.

At least 73 records · Page 4Linked to original sources

Identification and subcellular localization of a novel mammalian dynamin-related protein homologous to yeast Vps1p and Dnm1p.

The dynamin family of GTP-binding proteins are implicated in vesicular transport. These include mammalian dynamins I, II, III, and yeast Vps1p and Dnm1p. Dynamin is involved in the formation of clathrin-coated vesicles at the plasma membrane. On the other hand, Vps1p and Dnm1p appear to be involved in transport from the late Golgi compartment to vacuoles and in an endocytic process, respectively. In this study, we identified a novel human protein, named Dnm1p/Vps1p-like protein (DVLP). It resembled more closely Dnm1p and Vps1p than dynamins not only in the primary structure but also in the domain organization. DVLP mRNA was expressed ubiquitously, suggesting that this protein plays a fundamental role in cellular function. Immunofluorescence analysis of cells expressing epitope-tagged DVLP revealed that it showed a diffused perinuclear staining pattern that was not superimposed on that of the marker protein for the Golgi apparatus, trans-Golgi network, lysosomes, endosomes, or endoplasmic reticulum. These data suggest that DVLP is not involved in the formation of known coated vesicles.

Amino Acid Sequence↗

Topical administration of prostaglandin E1 with iontophoresis for skin flap viability.

This study was performed to investigate the capability of iontophoretic delivery of prostaglandin E1 (PGE1) and the effect of this treatment on the viability of skin flaps on the rat dorsum model as described by Hammond and Ronald in 1993. The PGE1 level in the tissue sample under the electrode was assessed with radioimmunoassay. The iontophoretic treatment (for 20 minutes at 4 mA) with PGE1 solution (20 micrograms PGE1 in 2 ml saline) showed a significant increase of PGE1 in the flap tissue under the negative electrode (p < 0.05). Very little increase was observed after the iontophoresis of saline alone. This was not statistically significant. Iontophoretic treatment was given for 5 consecutive days following flap elevation. The survival area on the seventh postoperative day was significantly greater in the PGE1-treated flaps than that of either the control or saline-treated flaps. These results confirm the beneficial effect of iontophoretic treatment with PGE1 on the augmentation of skin flap viability.

Administration, Topical↗

Surgical approach to midline skull base tumors with olfactory preservation.

Through the frontal approach, a skull base tumor in the deep midline region is obscured by the cribriform plate. This paper demonstrates our experience of operating on seven patients with midline skull base tumors posterior to the cribriform plate using a technique of maintaining the olfactory-cribriform anatomy. Following a bifrontal craniotomy and an orbitonaso-glabellar osteotomy, a circumferential osteotomy of the cribriform plate and horizontal incision of the nasal mucosa permitted mobilization of the cribriform plate unit along with the frontal lobe, providing a wide exposure of the posterior midline area. Tumors were resected from the posterior nasopharynx, ethmoid sinus, sphenoid sinus, and upper portion of the clivus. The skull base defects were repaired with the bipedicled temporoparietal galeal flap. Two patients lost olfaction because of poor preservation of the nasal mucosa and intradural dissection of the olfactory bulb. Five patients showed complete recovery of olfaction in 8 weeks. None of the patients developed cerebrospinal fluid leakage, infection, or increased neurologic deficits. En bloc mobilization of the cribriform plate and the bipedicled temporoparietal galeal flap are very useful for the management of midline cranial base tumors posterior to the cribriform plate as long as the olfactory unit is not involved with tumors.

Adolescent↗

A case retaining contact urticaria against egg white after gaining tolerance to ingestion.

A girl, 5.7 years old, gained tolerance to egg white ingestion in spite of high immunoglobulin E (IgE) antibody titers to egg white but retained contact urticaria against egg white. She developed atopic dermatitis on her face at 2 months of age and showed high IgE antibody titers to egg white and cow's milk. Accidental ingestion of egg products initiated immediate symptoms such as wheezing, urticaria, erythema and edema of the eyelids and conjunctiva three times. These symptoms were confirmed by challenge tests using boiled egg white at 3.9 years of age. She also reacted positively to a 20 min patch test on her volar arm with raw egg white. However, there were no reactions to the oral challenge test by boiled egg and freeze-dried egg white at 5.1 and 5.7 years of age, respectively. This non-responsiveness was confirmed by a double-blind, placebo-controlled food challenge using freeze-dried egg white. Nevertheless, she showed positive reactions to a 20 min patch test with freeze-dried egg white. Her IgE antibody titers to the egg white components including ovomucoid, ovalbumin, ovotransferrin and lysozyme as well as egg white were high from 2.9 to 5.7 years old. Her IgE antibody titers for the ovomucoid fragments digested by pepsin, chymotrypsin and trypsin were not lower than those of positive control subjects. The binding activity of IgE antibody to ovomucoid, however, decreased from 2.9 to 5.6 years as shown by radioallergosorbent test (RAST) inhibition assays. The IgE antibody showed weaker binding activity to pepsin- and chymotrypsin-digested ovomucoid that were filtered through cut-off 10,000 filter at the age of 2.1 and 5.7 years. We speculated that the maturation of secretion of digestive enzymes was involved in the mechanisms of the acquisition of tolerance to egg white ingestion in spite of the persistence of contact urticaria.

Antibody Affinity↗

Role of tachykinin and bradykinin receptors and mast cells in gaseous formaldehyde-induced airway microvascular leakage in rats.

We have investigated the effects of CP-99,994 [(+)-(2s,3s)-3-(2-methoxybenzylamino)-2-phenylpiperidine], a tachykinin NK1 receptor antagonist, HOE 140 (D-Arg[Hyp3,Thi5,D-Tic7,Oic8]bradykinin), a bradykinin B2 receptor antagonist, and ketotifen (4-(1-methyl-4-piperidylidene)4 H-benzo[4,5]cycloheptal[1,2-b]thiophen-10(9H)-one hydrogen fumarate), a histamine H1 receptor antagonist with mast cell-stabilizing properties, on microvascular leakage induced by gaseous formaldehyde. Extravasation of Evans blue dye into airway tissues was used as an index of airway microvascular leakage. Leakage of dye in the trachea and main bronchi increased significantly in a concentration-dependent fashion after 10 min inhalation of formaldehyde (5-45 parts per million (ppm)). The airway response induced by 10 min inhalation of 15 ppm formaldehyde (trachea: 119.5 +/- 13.9 ng/mg, n = 7; main bronchi: 139.6 +/- 7.9 ng/mg, n = 7) was abolished by the administration of CP-99,994 (3 and 6 mg/kg i.v.), but not by the administration of HOE 140 (0.65 mg/kg i.v.) nor ketotifen (1 mg/kg i.v.). The increase in vascular permeability induced by formaldehyde in the rat airway was mediated predominantly by NK1 receptor stimulation. Activation of bradykinin receptors and mast cells did not appear to play an important role in this airway response.

Animals↗

Role of bradykinin B2 receptors and mast cells in the bradykinin-induced skin response in the rat.

We investigated the role of activation of bradykinin receptors and mast cells in the microvascular leakage of the vessels of the skin induced by the intracutaneous (i.c.) injection of bradykinin in the rat. We evaluated the effects of HOE140 (D-Arg-[Hyp3,Thi5,D-Tic7,Oic8]bradykinin), a bradykinin B2 receptor antagonist, and ketotifen (4-(1-methyl-4-piperidylidene)4H-benzo[4,5]cyclohepta[1, 2-b]thiophen-10(9H)-one hydrogen fumarate), a histamine H1 receptor antagonist with mast cell stabilizing properties, on the skin response. Evans blue dye extravasation served as an index of the increase in vascular permeability. Bradykinin (2-100 nmol/site i.c.) induced the extravasation of Evans blue dye in a dose-dependent manner. Ketotifen (20 mg/kg i.p.) significantly inhibited the leakage of dye induced by bradykinin (10 nmol/site i.c.) by 66.2%, while HOE140 (1 mg/kg i.v.) had no effect. The concomitant injection of HOE140 (0.2, 2 nmol/site) and bradykinin (10 nmol/site i.c.), also did not significantly reduce the extravasation of dye. We conclude that the extravasation of plasma induced by the i.c. injection of bradykinin is mediated mainly by stimulation of the skin mast cells, but not by bradykinin B2 receptors.

Animals↗

Interleukin-2 receptor gamma-chain mutations in severe combined immunodeficiency with B-lymphocytes.

UNLABELLED: Severe combined immunodeficiency (SCID) with a normal number of B-lymphocytes usually demonstrates an X-linked inheritance and now is regarded as an interleukin-2-receptor (IL-2R) gamma-chain gene defect. Here, we report the characterization of mutations in the IL-2R gamma-chain gene of six unrelated SCID patients. One large deletion, one short deletion, one nonsense mutation and three single missense mutations were identified. The missense mutations were located near the motifs common to members of the class I cytokine receptor family. Two of the missense mutations were the same as previously reported in spite of the difference of ethnic backgrounds. The remaining four patients had newly identified mutations. CONCLUSION: Our results emphasize the broad molecular heterogeneity of X-linked SCID and suggest the presence of mutational "hot spots" within the IL-2R gamma-chain gene.

B-Lymphocytes↗

The reduction and redistribution of plantar pressures using foot orthoses in diabetic patients.

High pressures on the soles of the feet of diabetic patients cause plantar ulcerations, and reduction of this pressure is important in the treatment and prevention of such ulcers. The effects of foot orthoses on the distribution of plantar pressures were investigated using a pressure-sensitive insole containing 960 sensor elements in seven diabetic patients (13 feet) with diabetic ulcers or past histories of diabetic ulcers. Plantar pressures at the foot-insole interface and their distribution were measured in a stable standing position wearing the patients' own shoes, with and without foot orthoses. The preorthotic peak pressure was 130.6 +/- 41.9 kPA (mean +/- S.D.), while the postorthotic peak pressure was reduced to 52.6 +/- 17.9 kPa (P < 0.001 vs. preorthotic). The mean reduction of pressure was 56.3%. The preorthotic contact area was 330.2 +/- 48.4 pressure-sensitive elements and the postorthotic contact area was increased to 517.2 +/- 105.5 elements (P < 0.001), with a mean increase of 62.7%. The marked reduction in plantar pressures at the foot-insole interface by pressure redistribution indicates that this new pressure measurement system is very useful for the design of such orthoses.

Adult↗

Immunosuppression by anti-ICAM-1 and anti-LFA-1 monoclonal antibodies of free and vascularized skin allograft rejection.

Immunosuppression by anti-adhesion molecule antibody of free or vascularized skin allograft rejection was investigated in rats. Lewis (LEW, RT11) rats were used as donors and Fisher (F344, RT11v1) rats as the recipients. When F344 rats were treated intraperitoneally (i.p.) with anti-intercellular adhesion molecule-1 (ICAM-1) mAb (1A29) (3 mg/kg/day) and anti-leukocyte function-associated antigen-1 (LFA-1) mAb (WT.1) (3 mg/kg/day) one day prior to grafting and daily after grafting for nine days, free skin graft survival was prolonged only slightly compared with that in control rats which were injected i.p. with a daily dose of 6 mg/kg of anti-TNP mAbs (H1-6-2) one day prior to grafting and daily after grafting for nine days. (Mean survival time [MST] of the free skin graft was 11.2 +/- 0.6 days in the control group and 13.4 +/- 0.3 days in the 1A29 + WT-1 treated group [p < 0.01], respectively.) On the other hand, the vascularized graft survival was prolonged significantly in anti-ICAM-1/LFA-1 mAbs-treated F344 rats as compared with that in control rats. (The mean vascularized graft survival time was 14.2 +/- 0.7 days in the control group and 21.5 +/- 1.9 days in 1A29 + WT-1 treated group [p < 0.002]). Our results suggest that interaction with ICAM-1 and LFA-1 is more important in the rejection of vascularized skin allografts than that of free skin allografts.

Animals↗

Structural changes and cell viability of cultured epithelium after freezing storage.

Numerous clinical reports have shown the utility of cultured epithelial grafting in the field of plastic and reconstruction surgery. Recently, freezing storage of the cultured epithelium has been tried and has successfully grafted after thawing. It is clinically convenient if it is possible for cultured epithelium to keep its normal structure and viability. However, few papers have described the structural changes in cultured epithelium after freezing storage. In the present study, the morphological changes and cell viability of cultured mucosal epithelial sheets after freezing were studied in comparison with cultured epidermal sheets. Furthermore, we discuss the effect of storage temperature and cryoprotectants. As a result, there were some structural changes such as vacuolar degeneration in the cultured mucosal sheets using dimethyl sulphoxide (DMSO) as a cryoprotectant. Such changes were more clearly observed at -80 degrees C than at -196 degrees C with DMSO. However, little morphological change was observed in both epithelial sheets cultured with glycerin. The cell viability analysed by flow cytometry showed that more than 62% of the cells kept their viability after freezing storage. These results suggest that the optimum conditions of freezing for cultured epithelium were -196 degrees C storage by slow cooling methods with glycerin as a cryoprotectant.

Cell Survival↗

Transplantation of cultured mucosal epithelium: an experimental study.

We investigated morphological changes after transplantation of cultured mucosal epithelium using a modified Barrandon's method (1988). Serially cultivated human mucosal epithelium was transplanted onto the reverse side of rectangular dorsal skin flaps in hairless mice. The morphological changes in the epithelium were studied using paraffin sections. The modified Barrandon's method used in this study has advantages such as minimum external trauma and less chance of infection. The cultured epithelium was taken within 1 week and gradually increased its epithelial thickness. Keratinized epithelium arises after 3 weeks. At 4 weeks after grafting, the grafted epithelium comprised 7-10 cell layers. The structure of transplanted tissue, in conjunction with surrounding connective tissues, showed dermis-like features at day 7 after transplantation. From these results, it was confirmed that cultured mucosal epithelium could be successfully transplanted and its morphology was similar to that of normal mucosal tissue.

Animals↗

Structure and intracellular localization of mouse ADP-ribosylation factors type 1 to type 6 (ARF1-ARF6).

ADP-ribosylation factors (ARFs) are a family of small GTP-binding proteins that are proposed to be involved in the formation of coated transport vesicles. Although six ARF sequences have been reported in mammals to date, it has been unclear how many ARF members are present in a single organism. In this study, we provide the first direct evidence by cDNA cloning for the presence of all six ARF members in mouse. These proteins are highly conserved across mammalian species and Northern blot analysis revealed that mRNAs for all the members were expressed ubiquitously. Transfection of cells with epitope-tagged ARFs revealed that ARFs 1-3 displayed a perinuclear Golgi localization, while ARFs 4-6 appeared to be widely dispersed throughout the cytoplasm. These results suggest that although all the ARF proteins play fundamental and critical roles in cellular function, they are involved in different vesicular transport processes.

ADP-Ribosylation Factor 1↗

Healing of microvenous polytetrafluoroethylene (PTFE) prostheses implanted into the rat femoral vein by means of 3M precise.

Thirty PTFE prostheses (internal diameter, 1.0 mm; length, 5.0 mm; fibril length, 30 microns) were implanted into rats' femoral veins by means of a coupling device (3M precise microvascular anastomosis system, 3M, St Paul, MN) and evaluated with electron microscopy at regular intervals from 1 day to 3 weeks after implantation to study, in detail, the healing process. Eighty-three percent of mechanically anastomosed grafts were found to be patent. At 1 and 3 days after implantation, the whole length of PTFE was covered with a clot layer containing platelets and a fibrin network. After 1 week, endothelial-like cells originating from the anastomotic sides grew in across the anastomoses. At 3 weeks, the prostheses were completely covered by an endothelial-like cell layer. These results demonstrate that the degree of neo-endothelialization in the microvenous PTFE prosthesis anastomosed with 3M rings was not delayed, as was seen in the healing of microarterial PTFE tubing that was mechanically anastomosed.

Animals↗

[The evaluation of the bio-compatibility and the clinical usefulness of heparin-coated cardiopulmonary bypass circuits].

We studied the biocompatibility and the clinical usefulness of heparin-coated cardiopulmonary bypass circuits (Duraflow-II) compared with non-coated circuits. First study was done to clarify the biocompatibility of heparin coated circuit. 33 cases of elective coronary artery bypass grafting were randomized into two grops. Both group had full dose heparin in this study and laboratory tests were done such as blood cell count, free hemoglobin, AT-III, fibrinogen, FDP, and complement system. The use of heparin-coated circuits resulted in a reduction of C3a generation, and a reduction of fibrinogen consumption. The decline of AT-III may be due to bonding of coated heparin to AT-III, leading to effective anticoagulation. Next study was carried out in 26 cases. In heparin coated group, the amount of heparin was reduced to 200 IU/kg compared to 350 IU/kg in control group. ACT was maintained above 300 sec. and 400 sec. respectively. The amount of post operative bleeding was identical in both groups. There was no case which required autologous blood transfusion, re-operation for bleeding in both groups. Myocardial infarction and hospital mortality were not seen in this study. Concerning the fear of graft occlusion in low dose heparin surgery, there was no statistical difference of graft patency in both groups. In conclusion, heparin coated CPB circuits (Duraflo-II) are favorable in the meaning of biocompatibility and sefely used with low dose heparin.

Biocompatible Materials↗

[Early and mid-term results of right gastroepiploic artery grafting in the children with Kawasaki disease].

Thirty-nine children with Kawasaki disease have undergone coronary revascularization since 1976. The attrition rate of saphenous vein grafts (SVG) was so high that we have used the internal thoracic artery (ITA) as the graft since 1984. ITA showed excellent early and mid-term patency. We, however, still had to use SVG for the distal portion of right coronary artery (RCA) and left circumflex artery (LCx) where ITA was difficult to reach. In 1988, we first successfully applied the right gastroepiploic artery (GEA) for revascularization to the posterior descending artery of a six-year-old boy. Nine children were then bypassed with the combination of ITA and GEA. GEA was anastomosed to the distal portion of RCA (7 cases) and the branch of LCx (2 cases). In 4 cases GEA showed string sign 1 month after surgery, but the angiographic finding revealed improvement after 1 year interval in 2 cases. The patency of GEA grafts was excellent after 1 month (100%:9/9) and 1 year (100%:9/9), respectively. The use of GEA in addition to ITA as the arterial graft may contribute to the improvement of long-term graft patency in revascularization of children with Kawasaki disease.

Adolescent↗

Cytotoxicity of brefeldin A correlates with its inhibitory effect on membrane binding of COP coat proteins.

The fungal metabolite brefeldin A (BFA) causes the inhibition of protein secretion and the disruption of the structure and function of organelles along the exocytic and endocytic pathways including the Golgi complex. Such effects of BFA have been ascribed in large part to its ability to prevent recruitment of cytosolic coat proteins onto organelle membranes. Here we show that mammalian cell lines differ from one another with respect to sensitivity to this drug. The BFA sensitivity of a given cell line appears to be dependent on the species or the order from which the cell line originates, rather than on the cell line itself. In each cell line, the dose of BFA required for inhibition of cell growth and of protein secretion correlates with the dose required for inhibition of binding of beta-COP, a coat protein of COP-coated vesicles, but not that for inhibition of binding of gamma-adaptin, a component of HA-I/AP-1 adaptor of clathrin-coated vesicles. These observations suggest that: (i) there are at least two targets for BFA that differ from each other in sensitivity to this drug, (ii) the difference in the sensitivity to BFA of the beta-COP binding is determined by the difference in the structure of a target protein for this drug, and (iii) the cytotoxicity of BFA is ascribed mainly to its inhibitory effect on the membrane binding of COP-coat proteins.

3T3 Cells↗

The characteristics of cultured mucosal cell sheet as a material for grafting; comparison with cultured epidermal cell sheet.

The characteristics of cultured mucosal cells from the oral mucosa were investigated and compared with those of cultured epidermal cells. Total cell counts showed that mucosal cells possessed greater proliferating ability than epidermal cells. The results of 3(4,5-dimethyle-thiazoyl-2-yl)2,5 diphenyltetrazolium bromide assay confirmed this observation and also suggested that the mucosal cells maintained biological activity longer than epidermal cells. The most important morphological characteristics of mucosal cells in culture were their low grade of differentiation. Interestingly, the epidermal cells showed enucleation and keratinization progressively during culture, whereas the mucosal cells showed no obvious enucleation when examined by light microscopy. Transmission electron microscopy showed a smaller number of desmosomes in cultured mucosal cells than epidermal cells. The results of this study reveal cultured mucosal cell sheets to be a possible material for grafting in addition to cultured epidermal cell sheets.

Cell Count↗