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Biomedical subjects

K Shimizu

Publications and source records attributed to K Shimizu.

At least 649 records · Page 36Linked to original sources
PubMed1996

Different effects of various phospholipids on Ki-Ras-, Ha-Ras-, and Rap1B-induced B-Raf activation.

We have recently purified a Ki-Ras- and Ha-Ras-dependent extracellular signal-regulated kinase kinase from bovine brain and identified it as B-Raf protein kinase complexed with 14-3-3 proteins (Yamamori, B., Kuroda, S., Shimizu, K., Fukui, K., Ohtsuka, T., and Takai, Y. (1995) J. Biol. Chem. 270, 11723-11726). Moreover, we found that Rap1B as well as Ki-Ras and Ha-Ras stimulate the B-Raf activity. Since B-Raf contains a cysteine-rich domain originally found in protein kinase C as a domain responsible for interaction with phosphatidylserine (PS) and diacylglycerol or 12-O-tetradecanoylphorbol-13-acetate, we have examined here the effect of these compounds on the Ki-Ras-, Ha-Ras-, and Rap1B-induced activation of bovine brain B-Raf. Bovine brain PS enhanced Ki-Ras-stimulated B-Raf activity. Phosphatidic acid was slightly active, but other phospholipids, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol (PI), PI-4-monophosphate, PI-4,5-bisphosphate, and PI-3,4,5-trisphosphate, were inactive. However, none of the above phospholipids affected the Ha-Ras-stimulated B-Raf activity, whereas PI, PS, phosphatidylethanolamine, and phosphatidic acid inhibited the Rap1B-stimulated B-Raf activity. Phosphatidylcholine or PI-4-monophosphate did not show any effect on the Rap1B-stimulated B-Raf activity. Synthetic PS with two unsaturated fatty acids, such as 1,2-dioleoyl-PS or 1,2-dilinoleoyl-PS, showed the same effect toward the Ki-Ras- and Rap1B-stimulated B-Raf activities, but synthetic PS with two saturated fatty acids, such as 1, 2-distearoyl-PS, was inactive. 12-O-Tetradecanoylphorbol-13-acetate did not affect the stimulatory or inhibitory effect of PS on the Ki-Ras- and Rap1B-stimulated B-Raf activities, respectively. PS did not affect the Ki-Ras-, Ha-Ras-, or Rap1B-independent basal B-Raf activity or the mitogen-activated protein kinase kinase or extracellular signal-regulated kinase activity. These results indicate that various phospholipids differently affect Ki-Ras-, Ha-Ras, and Rap1B-induced B-Raf activation.

Animals↗
PubMed1996

Immunosuppressant FK506 induces interleukin-6 production through the activation of transcription factor nuclear factor (NF)-kappa(B). Implications for FK506 nephropathy.

FK506 is a powerful immunosuppressive drug currently in use that inhibits the activation of several transcription factors (nuclear factor (NF)-AT and NF-kappaB) critical for T cell activation. We show here that, contrary to the situation in T cells, FK506 activates transcription factor NF-kappaB in nonlymphoid cells such as fibroblasts and renal mesangial cells. We further show that FK506 induces NF-kappaB-regulated IL-6 production in vitro and in vivo, in particular in kidney. IL-6 has been shown previously to produce renal abnormalities in vivo, such as mesangioproliferative glomerulonephritis. Similar renal abnormalities were also observed in FK506-treated animals. These results thus suggest a causal relationship between FK506-induced NF-kappaB activation/IL-6 production and some of FK506-induced renal abnormalities.

Animals↗
PubMed1996

Activation of brain B-Raf protein kinase by Rap1B small GTP-binding protein.

Rap1 small GTP-binding protein has the same amino acid sequence at its effector domain as that of Ras. Rap1 has been shown to antagonize the Ras functions, such as the Ras-induced transformation of NIH 3T3 cells and the Ras-induced activation of the c-Raf-1 protein kinase-dependent mitogen-activated protein (MAP) kinase cascade in Rat-1 cells, whereas we have shown that Rap1 as well as Ras stimulates DNA synthesis in Swiss 3T3 cells. We have established a cell-free assay system in which Ras activates bovine brain B-Raf protein kinase. Here we have used this assay system and examined the effect of Rap1 on the B-Raf activity to phosphorylate recombinant MAP kinase kinase (MEK). Recombinant Rap1B stimulated the activity of B-Raf, which was partially purified from bovine brain and immunoprecipitated by an anti-B-Raf antibody. The GTP-bound form was active, but the GDP-bound form was inactive. The fully post-translationally lipid-modified form was active, but the unmodified form was nearly inactive. The maximum B-Raf activity stimulated by Rap1B was nearly the same as that stimulated by Ki-Ras. Rap1B enhanced the Ki-Ras-stimulated B-Raf activity in an additive manner. These results indicate that not only Ras but also Rap1 is involved in the activation of the B-Raf-dependent MAP kinase cascade.

3T3 Cells↗
PubMed1996

Possible involvement of RGD (Arg-Gly-Asp)-containing extracellular matrix proteins in rat growth plate chondrocyte differentiation in culture.

RGD (arg-gly-asp)-containing proteins have been shown to be components of cartilage matrix. In the present study, the role of interactions of cells with RGD-containing cartilage matrix proteins in rat costal epiphyseal chondrocyte differentiation was examined using a pelleted culture system as an in vitro model of endochondral ossification. Cell attachment assays showed the presence of integrins which mediated the binding of chondrocytes to fibronectin, a member of RGD-containing cartilage matrix proteins, in an RGD-dependent manner. In the early culture period, when chondrocytes had nonhypertrophic morphology with low levels of alkaline phosphatase, the exogenous addition of synthetic peptide GRGDSP (gly-arg-gly-asp-ser-pro) caused an increase in alkaline phosphatase levels and enlargement of chondrocytes in pelleted cultures. Treatment with GRGDSP from the early to late culture periods in association with the transition of chondrocytes from prehypertrophic to hypertrophic phenotypes followed by matrix mineralization resulted in suppression of mineral growth without significant effects on alkaline phosphatase levels or cellular morphology in the cultures. Similarly, addition of the synthetic peptide during the late culture period with the advance of cartilage mineralization suppressed mineral growth in pelleted cultures. These data indicate an important role of interactions of chondrocytes with RGD-containing cartilage matrix proteins through integrins in the regulation of epiphyseal chondrocyte differentiation in pelleted cultures.

Amino Acid Sequence↗
PubMed1996

Suction/irrigation for deep wound infection after spinal instrumentation: a case study.

Deep wound infection after spinal instrumentation is a serious complication that is difficult to treat without removing the instruments and bone graft. Debridement and suction/irrigation is an effective method of treatment in these cases. It was performed on six patients in our department who developed this complication between 1985 and 1994. Four patients with early post-operative infection were cured by this method without removing the instruments and bone graft, and two patients with delayed post-operative infection were cured by this method with instrument removal. Debridement and suction/irrigation is a useful method of treatment for both groups of deep wound infection and gives good results when performed soon after infection onset together with additional antibiotic therapy.

Adult↗
PubMed1996

The immunosuppressive effects of a leukotriene B4 receptor antagonist on liver allotransplantation in rats.

The immunosuppressive effects of a leukotriene B4 (LTB4) receptor antagonist, ONO4057, on liver allotransplantation in rats were evaluated, and the levels of prostaglandin E2 (PGE2) in the liver tissue during rejection of the allografts examined. The rats were divided into four groups: group 1: Lewis rats (LEW) given a sham operation with dimethyl sulfoxide (DMSO); group 2: LEW given syngenic orthotopic liver transplantation (OLT) from LEW, with DMSO; group 3: LEW given allogenic OLT from ACI rats (ACI), with DMSO; and group 4: LEW given allogenic OLT from ACI, with ONO4057 as 10, 30, or 100 mg/kg per day dissolved in DMSO to subgroups 4a, 4b, and 4c, respectively. Histological examinations were performed, survival times monitored, and liver tissue PGE2 levels 3, 5, 7, and 14 days after transplantation measured. The mean graft survival times in groups 4a, b, and c, at 37.5 +/- 10.4, 52.2 +/- 24.4, and 34.0 +/- 4.9 days (mean +/- SEM), respectively, were significantly longer than that in group 3 (at 13.0 +/- 3.2 days). Moreover, the levels of tissue PGE2 in the liver allografts in group 4a were significantly higher than those in group 3 on days 5 and 7. These results suggest that ONO4057 has an immunosuppressive effect on liver allotransplantation since it reduces the activities of LTB4 which augments immune responses, and also because it indirectly increases the PGE2 level.

Alanine Transaminase↗
PubMed1996

Socioeconomic and behavioural risk factors for mortality of individuals with IDDM in Japan: population-based case-control study. Diabetes Epidemiology Research International (DERI) US-Japan Mortality Study Group.

The aim of this population-based, one-to-one matched-pair case-control study was to evaluate the factors concerning the markedly increased risk for dying among Japanese subjects with insulin-dependent diabetes mellitus (IDDM) from a social and behavioural perspective. The study was based on the population-based cohort of IDDM subjects in the Diabetes Epidemiology Research International Mortality Study. We studied 90 cases who died and 90 living control subjects, selected from the rest of the cohort, who were matched for sex, birth year, year of diagnosis and duration of diabetes. Socioeconomic and behavioural status were surveyed through a questionnaire. Conditional logistic regression analyses based on 55 respondent pairs revealed that the better educated patients (year of completing education: odds ratio = 0.66) who kept the same physician (number of times a patient changed physician: odds ratio = 2.77) and who attended a clinic specializing in diabetes (attendance at university hospital clinic: odds ratio = 0.18) injecting insulin several times a day (number of injections, odds ratio = 0.31) and more frequently attending the clinic (> or = 12 times per year, odds ratio = 0.23) were at substantially lower risk of death. The results begin to profile the patients with the highest risk of dying who could be identified earlier and undergo intervention treatment.

Adolescent↗
PubMed1996

Interleukin-6 (IL-6) production in carcinoma of the cervix.

Interleukin-6 (IL-6) is a pleiotropic cytokine that is not only a mediator in major immunologic reactions but also a growth factor of keratinocytes. We studied the IL-6 secretion in vitro of 15 human cell lines derived from both squamous cell carcinoma (SCC) and adenocarcinoma of the uterine cervix. Four of the eight well differentiated SCC secreted a large amount (> 1500 pg/48 h/10(6) cells) of IL-6 in nude mice. In contrast, poorly differentiated SCC cell lines and all of the 7 adenocarcinoma cell lines secreted a small amount (< 500 pg/48 h/10(6) cells of IL-6). The expression of IL-6 mRNA of the cell lines correlated well with their IL-6 secretion potential. However, the expression of IL-6 receptor did not correlate with the IL-6 secretory potential. We also studied the IL-6 secretion of freshly isolated normal squamous epithelium and of dysplastic epithelium. In culture, two normal squamous epithelia secreted a large amount (> 2000 pg/48 h/10(6) cells), whereas 8 dysplasia epithelia secreted an extremely small amount (< 10 pg/48 h/10(6) cells). About one-third of patients with SCC had a raised serum IL-6 value. IL-6 production may help to differentiate between SCC and adenocarcinoma of the uterine cervix. IL-6 regulation seems to change in the course of SCC carcinogenesis.

Adenocarcinoma↗
PubMed1996

Lactoferrin-mediated protection of the host from murine cytomegalovirus infection by a T-cell-dependent augmentation of natural killer cell activity.

The administration of bovine lactoferrin (LF) with 1 mg/g body weight before the murine cytomegalovirus (MCMV) infection completely protected the BALB/c mice from death due to the infection. In these LF-treated mice, a significant increase in the activity was found in the NK cells but not in the cytolytic T lymphocytes which recognized an MCMV-derived peptide. Moreover, the elimination of the NK cell activity by an injection with anti-asialo GM1 antibody abrogated such augmented resistance, thus supporting the hypothesis that the LF-mediated antiviral effect in vivo is performed through the augmentation of NK cell activity. No such LF-mediated antiviral effect in vivo with the increased NK cell activity was found in athymic nude mice, whereas it was restored completely by the transfer of splenic T cells from LF-treated donors. These findings therefore suggest that T lymphocytes induce both the augmentation of NK cell activity and the resultant antiviral effect in the LF-treated hosts.

3T3 Cells↗
PubMed1996

An angiomyoma with t(X;10)(q22;q23.2).

A t(X;10)(q22;q23.2) translocation was detected as the only chromosomal aberration in primary short-term cultured cells from an angiomyoma of a 58-year-old woman; 6p, 13q, and 21q rearrangements have been previously demonstrated by others in two cases of angiomyoma. This type of translocation has not been reported in other benign tumors including leiomyomas and angiomyomas, although it has been detected in an ependymoma. This is thought to be a third case of angiomyoma showing karyotypic abnormalities.

Angiomyoma↗
PubMed1996

Human papillomavirus type 16 E6 protein up-regulates the expression of the high mobility group protein HMG-I(Y) gene in mouse 10T1/2 cells.

Using a differential hybridization technique, we have identified a mouse cellular gene, high mobility group protein HMG-I(Y), whose expression is up-regulated by the E6 protein of human papillomavirus (HPV) type 16. This gene was overexpressed in E6-expressing mouse 10T1/2 cells, but not in G418-resistant 10T1/2 cells. The expression of the HMG-I(Y) gene was up-regulated by the transient expression of E6 from a zinc-inducible human metallothionein-IIA gene promoter. Expression was found to be more efficient at a confluent cell density than at a subconfluent cell density. The up-regulation of HMG-I(Y) gene expression by E6, in particular at a confluent cell density, may be part of an altered genetic program in host cells infected with HPV-16.

Animals↗
PubMed1996

Identification and partial characterization of vitellin from the barnacle Balanus amphitrite.

The major protein in occytes of the barnacle Balanus amphitrite has been identified as vitellin (Vt) and its biochemical properties partially characterized. SDS-PAGE of protein extracts from different developmental stages of occytes and embryos of B. amphitrite revealed that a 170 kDa polypeptide was abundantly present in fully matured oocytes. Its amount increased with occyte maturation and decreased with embryonic development after fertilization. Antiserum, prepared against 170 kDa polypeptide, was found to recognize only 170 kDa among proteins in ovary extract (OE) by immunoblot analysis. Immunohistochemical study indicated localization of 170 kDa polypeptide in yolk granules of oocytes, oviducts and embryos. Combination of native electrophoresis with agarose gel and SDS-PAGE displayed that the main protein in OE was lipophilic, contained 85 kDa polypeptide in addition of 170 kDa, and was positive in PAS reaction. The main peak eluted from OE contained 170 kDa and 85 kDa polypeptides, and corresponded to 200 kDa when filtrated through superose 6 gel. Conclusively, vitellin of B. amphitrite is a lipophilic glycoprotein consisting of two subunits, a heavy chain with 170 kDa and a light chain with 85 kDa. Two other barnacles, Megabalanus rosa and Tetraclita squamosa japonica, also contained proteins similar to B. amphitrite Vt.

Animals↗
PubMed1996

Bilateral lesions of the hypothalamic suprachiasmatic nucleus eliminated sympathetic response to intracranial injection of 2-deoxy-D-glucose and VIP rescued this response.

We previously found that bilateral lesions of the suprachiasmatic nucleus abolished hyperglycemic response to intracranial injection of 2-deoxy-D-glucose in rats. Because the hyperglycemia due to 2-deoxy-D-glucose was shown to be dependent on the functions of the adrenal medulla and sympathetic nervous system, the effect of bilateral lesions of the suprachiasmatic nucleus on changes in the nervous activity of sympathetic efferents to the adrenal after intracranial injection of 2-deoxy-D-glucose was examined in rats. It was found that bilateral lesions of the nucleus eliminated the increase in neural activity of the sympathetic efferent that occurred after the injection of 2-deoxy-D-glucose. Because the suprachiasmatic nucleus possesses neurons containing a vasoactive intestinal polypeptide-like substance, the effects of vasoactive intestinal polypeptide and 2-deoxy-D-glucose, administered alone or in combination, on the sympathetic activity were examined in intact control rats and in rats with bilateral lesions of the suprachiasmatic nucleus. It was found that in the normal control rats, vasoactive intestinal polypeptide alone increased the sympathetic activity, whereas it dramatically enhanced the sympathetic response to 2-deoxy-D-glucose. However, in rats with bilateral lesions of the suprachiasmatic nucleus, vasoactive intestinal polypeptide alone elicited no increase in the nervous activity of the sympathetic efferents to the adrenal, but combined administration of vasoactive intestinal polypeptide and 2-deoxy-D-glucose caused an increase in the nervous activity of sympathetic efferents to the adrenal. These findings suggest that the suprachiasmatic nucleus is involved in the enhancement of sympathetic activity caused by intracranial injection of 2-deoxy-D-glucose, and that neurons containing a vasoactive intestinal polypeptide-like substance in the suprachiasmatic nucleus play an important role in the sympathetic enhancement that occurs after intracranial injection of 2-deoxy-D-glucose. This role might be a permissive and facilitative one.

Analysis of Variance↗
PubMed1996

Four cases of selenium deficiency in postoperative long-term enteral nutrition.

Because selenium is seldom added to formulations for enteral nutrition (EN), postoperative patients who are supported with EN are at risk for selenium deficiency. This report describes four cases of suspected selenium deficiency in long-term EN. Two patients underwent pancreaticoduodenectomy, one underwent total gastro-pancreatectomy, and one underwent esophageal resection and reconstruction with jejunal autotransplantation. They all developed malabsorption syndrome within 2 yr after operation. Enteral nutritional support with an elemental diet was provided continuously for 7-11 yr. Over the past 1-2 yr they experienced increasing bilateral muscular pain and weakness in the legs, gait disturbance, palpitation, and shortness of breath. Investigation for possible trace element deficiency revealed very low levels of selenium in the blood. After 10-20 d of supplementation with daily intravenous administration of selenious acid 0.16 mg/d (100 micrograms/d of selenium), their blood levels of selenium rose and their symptoms resolved. They were then continued on a maintenance regimen of oral sodium selenite 0.13 mg/d (60 micrograms/d of selenium).

Aged↗
PubMed1996

The role of the premacular liquefied pocket and premacular vitreous cortex in idiopathic macular hole development.

PURPOSE: To clarify the role of the vitreous in idiopathic macular hole formation. METHODS: We prospectively evaluated the vitreous before and during vitreous surgery in 64 consecutive eyes of 62 patients (stage 1B, four eyes; stage 2, ten eyes; stage 3, 43 eyes; stage 4, four eyes) with macular holes; another three eyes underwent a second vitrectomy. After core vitrectomy, the premacular vitreous cortex was examined by applying gentle suction. The detached posterior hyaloid face was observed under endoilluminiation. RESULTS: We observed in all eyes a liquefied lacuna anterior to the posterior fundus before and during vitreous surgery. The premacular vitreous cortex was extremely thin and elastic upon gentle suction. A premacular ring, the diameter two to four times that of the Weiss ring, was observed in 48 of 57 eyes (84%) with stages 1B, 2, and 3 macular hole. In most cases, the vitreous cortex within the premacular ring either was absent or had a break, resulting in a premacular round defect. The vitreous cortex peripheral to the premacular ring, which was thick and less deformed, never showed a break. In eyes that had undergone a second vitrectomy, we noted residual cortex or an epiretinal membrane around the macular hole. CONCLUSIONS: The premacular vitreous cortex is extremely thin and elastic. It is sharply demarcated by a ring from the thick peripheral vitreous cortex. Tangential traction, which causes macular holes, appears to originate exclusively in the premacular vitreous cortex that forms the posterior wall of the premacular liquefied pocket.

Eye Diseases↗
PubMed1996

Insulin increases distinct species of 1,2-diacylglycerol in isolated perfused rat heart.

Insulin and glucose increase the synthesis of 1,2-diacylglycerol (1,2-DAG), the physiological activator of protein kinase C (PKC) in a variety of tissues and cells. The effects of insulin and glucose on the abundance and fatty acid composition of 1,2-DAG were investigated in isolated perfused rat hearts with the use of capillary gas chromatography and 1,2-dipentadecanoin as an internal standard. A high concentration of insulin (25 mU/ mL) significantly increased cardiac contractility and reduced coronary flow. In addition, perfusion with 25 mU/mL insulin induced significant increases of 18.2% and 26.4% in 1,2-DAG mass after 5 and 30 minutes, respectively, in the presence of 8.6 mmol/L glucose, whereas there was no increase in 1,2-DAG with 2.5 mU/mL insulin. Analysis of the fatty acid composition of 1,2-DAG showed that only species containing specific fatty acids (16:0, 18:1, and 18:2) were increased in response to insulin. In contrast, an increase in glucose concentration in the perfusion medium from 3 to 17 mmol/L had no effect on the total mass or fatty acid composition of 1,2-DAG, cardiac contractility, or coronary flow. Addition of a high insulin concentration to the high-glucose medium increased the abundance of 1,2-DAG containing 16:0, 18:1, and 18:2 fatty acids, as well as cardiac contractility. It is concluded that the effect of insulin on cardiac contractility may be related to the associated increase in 1,2-DAG abundance.

Animals↗
PubMed1996

Watershed zone in the human peripheral choroid.

PURPOSE: To evaluate the angioarchitecture and the hemodynamics in the peripheral choroid in humans. METHODS: Choroidal circulation was examined in 23 normal eyes and in 7 with equatorial lattice degeneration by wide-angle indocyanine green (ICG) angiography using a scanning laser ophthalmoscope. A hand-held 30-diopter lens was placed in front of the eye to expand the observation angle from the original 40 degrees to 70 degrees. The peripheral temporal choroid and the ora serrata thus could be seen in one frame. RESULTS: An arterial watershed zone was identified in the temporal peripheral choroid in 21 of 30 eyes. This watershed zone was located in a sectorial region posterior to the ora serrata. It did not form a circular belt along the equator. The area anterior to the watershed zone was supplied by peripheral retrograde choroidal arteries. Inflow of dye into the peripheral retrograde choroidal arteries was consistently later than into the more posterior choroid via short posterior ciliary arteries. The terminal branches of peripheral and short posterior ciliary arteries did not form functional anastomoses. Peripheral retrograde choroidal arteries showed wide variations in number and distribution. These arteries originated from recurrent branches of long posterior ciliary arteries in 18 of 21 eyes and from anterior ciliary arteries in 3 eyes. There was no correlation between the watershed zone and equatorial lattice degeneration in location and extent. CONCLUSION: A peripheral watershed zone is a regular feature in the far temporal sector of the choroid. The far peripheral choroid anterior to the watershed zone was perfused by peripheral retrograde choroidal arteries which originated from the long posterior ciliary arteries and/or the anterior ciliary arteries.

Adolescent↗