Biomedical subjects

Masahiro Sakata

Publications and source records attributed to Masahiro Sakata.

28 records · Page 2Linked to original sources

PubMed2004

Induction of glucose transporter 1 expression through hypoxia-inducible factor 1alpha under hypoxic conditions in trophoblast-derived cells.

Glucose transporter 1 (GLUT1) plays an important role in the transport of glucose in the placenta. During early pregnancy, placentation occurs in a relatively hypoxic environment that is essential for appropriate embryonic development, and GLUT1 expression is enhanced in response to oxygen deficiency in the placenta. Hypoxia-inducible factor-1 (HIF-1)alpha is involved in the induction of GLUT1 expression in other cells. The present study was designed to test whether HIF-1alpha is involved in hypoxia-induced activation of GLUT1 expression using trophoblast-derived human BeWo and rat Rcho-1 cells as models. GLUT1 mRNA and protein expression were elevated under 5% O2 or in the presence of cobalt chloride, which has been shown to mimic hypoxia. Using rat GLUT1 (rGLUT1) promoter-luciferase constructs, we showed that this up-regulation was mediated at the transcriptional level. Deletion mutant analysis of the rGLUT1 promoter indicated that a 184 bp hypoxia-responsive element (HRE) of the promoter was essential to increase GLUT1 reporter gene expression in response to low-oxygen conditions. BeWo and Rcho-1 cells cultured under 5% O2 or with CoCl2 showed increased expression of HIF-1alpha protein compared with those cultured under 20% O2. To test whether this factor is directly involved in hypoxia-induced GLUT1 promoter activation, BeWo and Rcho-1 cells were transiently transfected with an HIF-1alpha expression vector. Exogeneous HIF-1alpha markedly increased the GLUT1 promoter activity from constructs containing the HRE site, while the GLUT1 promoter constructs lacking the HRE site were not activated by exogenous HIF-1alpha These data demonstrate that GLUT1 is up-regulated under 5% O2 or in the presence of CoCl2 in the placental cell lines through HIF-1alpha interaction with a consensus HRE site of the GLUT1 promoter.

Blotting, Northern↗

PubMed2003

Inhibition of phosphorylation of a forkhead transcription factor sensitizes human ovarian cancer cells to cisplatin.

The Forkhead family transcription factor FKHRL1 is an inducer of apoptosis in its unphosphorylated form and was recently reported to be a substrate of Akt kinase. We studied the roles of FKHRL1 in both cisplatin-resistant Caov-3 (a papillary adenocarcinoma cell line) and cisplatin-sensitive A2780 human ovarian cancer cell lines. Treatment of Caov-3 cells but not A2780 cells with cisplatin transiently stimulated the phosphorylation of FKHRL1. Transfection experiments revealed that a kinase inactive-mutant of Akt or a triple mutant (TM) of FKHRL1, in which all three of the putative Akt phosphorylation sites were converted to alanine, was unable to phosphorylate the FKHRL1 protein in cells treated with cisplatin. Because the phosphorylated form of FKHRL1 is known to be localized in the cytoplasm, we examined whether cisplatin-induced phosphorylation of FKHRL1 might have an effect on the subcellular distribution of FKHRL1. Cisplatin induced the localization of FKHRL1 in the cytoplasm in Caov-3 cells but not in A2790 cells. Moreover, cisplatin induced the association of 14-3-3 protein with phosphorylated-FKHRL1 in Caov-3 cells but not in A2790 cells. Because the unphosphorylated form of FKHRL1 binds the Fas ligand promoter, thereby inducing apoptosis, we further examined the effect of the phosphorylation status of FKHRL1 on the activity of the Fas ligand promoter in the presence of cisplatin. Transfection with the kinase-inactive mutant of Akt or TM of FKHRL1 induced the activity of the Fas ligand promoter in Caov-3 cells. Moreover, exogenous expression of TM of FKHRL1 in Caov-3 cells decreased the cell viability after treatment with cisplatin. Our findings suggest that cisplatin causes the phosphorylation of FKHRL1 via a phosphatidylinositol 3-kinase/Akt cascade, and inhibition of this cascade sensitizes ovarian cancer cells to cisplatin.

14-3-3 Proteins↗

PubMed2003

Lymphatic cardiac tamponade after open-heart surgery with liver dysfunction.

A 46-year-old female with alcoholic liver dysfunction was admitted for mitral regurgitation due to infective endocarditis. She underwent mitral valvuloplasty and resection of the vegetation without complication. After removal of the chest tube, late cardiac tamponade occurred and subsequently recurred. On the 64th day after mitral valvuloplasty, we performed redo median sternotomy with small laparotomy trying to reveal and repair injured lymphatic vessels in the pericardial space and successfully cured the leakage of lymph. The post reoperative course was uneventful and the patient was discharged 20 days after reoperation. We review a rare complication of recurrent cardiac tamponade of lymphatic leakage associated with liver dysfunction.

Balloon Occlusion↗

PubMed2003

Topographical relationships between the brainstem auditory and somatosensory evoked potentials and the location of lesions in posterior fossa stroke.

The topographical relationships between the location of brainstem lesions detected by magnetic resonance imaging and abnormality of brainstem auditory evoked potentials (BAEPs) and short-latency somatosensory evoked potentials (SSEPs) were studied in 57 patients with stroke in the posterior fossa. Abnormal BAEPs or SSEPs were associated with lesions involving the pontine tegmentum, and abnormal BAEPs also with lesions at the cerebellar peduncle. Absence of the V wave in BAEPs and N20 in SSEPs was associated with a localized overlapping area in the pontine tegmentum contralateral to stimulation. The overlapping area associated with loss of N20 coincided with the location of the medial lemniscus. Lesions widely involving the pontine tegmentum caused the disappearance of multiple waves in the BAEPs and SSEPs. Patients who entered prolonged coma or died had total loss of the III, IV, and V waves, bilateral absence to the contralateral response in BAEPs, or loss of N18 in SSEPs. The loss of N18 in SSEPs had a statistically significant correlation with bad outcome, which suggests the superiority of SSEPs for predicting the outcome of stroke and indicates the involvement of some system excluding the medial lemniscus in the generation of N18.

Adult↗

PubMed2002

Expression of the anaphylatoxin receptor C5aR (CD88) by human articular chondrocytes.

Although the complement system is implicated in the inflammatory process in arthritic diseases, a direct interaction between chondrocytes and complement has not been demonstrated. In this study, we investigated expression of the C5a receptor (C5aR) on chondrocytes of cartilage from patients with rheumatoid arthritis (RA), osteoarthritis (OA), and bone fracture as normal controls by reverse transcriptase polymerase chain reaction (RT-PCR), flow cytometry, and immunohistochemistry. The RT-PCR detected mRNA for C5aR in most or all of the tested samples (73% in OA, 100% in RA, 89% in normal). The FACS analysis revealed different expression ratios between individuals varying from 0.7% to 77.1%; however, expression ratios of C5aR were significantly higher in RA than in controls (26.0% in RA, 9.0% in OA, 6.9% in normal). The expression of C5aR was upregulated significantly by addition of IL-1beta in RA and normal samples but not in OA. In addition, the C5aR-positive chondrocytes were confirmed by immunohistochemistry. In conclusion, expression of C5aR and the effect of IL-1beta on the expression were different between RA and OA. The C5aR may contribute to chondrocyte metabolism and the pathogenesis of arthritis differently between in RA and OA.

Adult↗

PubMed2002

Aortic aneurysm and aortic regurgitation following aortic valve replacement due to Takayasu's arteritis.

A 26-year-old man who underwent aortic valve replacement for aortic regurgitation due to Takayasu's arteritis 2 years earlier experienced left amaurosis persisting for some minutes. Computed tomography showed aneurysmal dilation of the ascending aorta to a diameter of 60 mm and occlusion of the left carotid artery. Cardiac echography showed perivalvular leakage. Following administration of a calcium antagonist, the patient's amaurosis subsided and brain bloodstream scintigraphy showed no abnormalities. We resected the aneurysm instead of using Bentall's operation. Following an uncomplicated postoperative course, the patient was discharged 21 days after surgery and echocardiography has shown no perivalvular leakage to date.

Adult↗

PubMed2002

Modified Ross procedure with stentless bioprosthesis and pericardial xenograft for the right ventricular outflow tract. Usefulness of autologous pericardial patch for hemostasis.

A 31-year-old man with severe aortic regurgitation due to a defective bicuspid valve underwent surgery using modified Ross procedure. The right ventricular outflow tract (RVOT) was reconstructed with a 25 mm stentless xenograft valve sutured with a rolled equine pericardium. Oozy bleeding from the RVOT was controlled with an autologous pericardial patch and fibrin glue. Postoperative echocardiography showed no aortic regurgitation. No blood transfusion was required.

Adult↗

PubMed2002

Expression and regulation of 4F2hc and hLAT1 in human trophoblasts.

The neutral amino acid transport system L is a sodium-independent transport system in human placenta and choriocarcinoma cells. Recently, it was found that the heterodimer composed of hLAT1 (a light-chain protein) and 4F2 heavy chain (4F2hc), a type II transmembrane glycoprotein, is responsible for system L amino acid transport. We found that the mRNAs of 4F2hc and hLAT1 were expressed in the human placenta and a human choriocarcinoma cell line. The levels of the 4F2hc and hLAT1 proteins in the human placenta increased at full term compared with those at midtrimester. Immunohistochemical data showed that these proteins were localized mainly in the placental apical membrane. Data from leucine uptake experiments, Northern blot analysis, and immunoblot analysis showed that this transport system was partially regulated by protein kinase C and calcium ionophore in the human choriocarcinoma cell line. Our results suggest that the heterodimer of 4F2hc and hLAT1 may play an important role in placental amino acid transport system L.

Calcium↗

PubMed2002

Autoimmunity against YKL-39, a human cartilage derived protein, in patients with osteoarthritis.

OBJECTIVE: Our previous study revealed that some patients with rheumatoid arthritis (RA) possessed autoantibodies to YKL-39, a cartilage related protein. We investigated whether patients with osteoarthritis (OA) also displayed autoimmunity to YKL-39. METHODS: Autoantibodies to recombinant YKL-39 as well as human cartilage glycoprotein-39 were detected by ELISA and Western blotting. The tested serum samples were derived from 117 patients with OA, 94 patients with RA, and 2 groups of 50 arthropathy-free healthy donors who matched the OA and RA groups for age and sex. We determined autoepitopes on YKL-39 using 3 overlapping fragments of YKL-39 (designated F1, F2, F3). T cell proliferation response to YKL-39 was analyzed using the 3H-thymidine incorporation assay. RESULTS: Autoantibodies to YKL-39 were detected in 13 (11.1%) patients with OA and 11 (11.8%) with RA. In the epitope mapping, all the 3 fragments of YKL-39 were found to carry autoepitopes, but F1 was recognized most frequently. Proliferative responses of peripheral blood mononuclear cells against YKL-39 were detected in 6 (46%) of the 13 OA patients who were positive for the anti-YKL-39 autoantibodies and in 2 (17%) of the 11 antibody positive RA patients. CONCLUSION: These results show that autoimmunity to YKL-39 in patients with OA was present at equal or somewhat higher frequency than in patients with RA. The cellular and humoral immune responses to YKL-39 may be involved in the pathological process of OA as well as RA.

Adipokines↗

PubMedDate not supplied

Estrogen and raloxifene inhibit the monocytic chemoattractant protein-1-induced migration of human monocytic cells via nongenomic estrogen receptor alpha.

OBJECTIVE: To investigate the effects of estradiol (E2) and raloxifene on the migration of human monocytic THP-1 cells to endothelium. DESIGN: A prospective comparative study. THP-1 cells, a human acute monocytic leukemia cell line, were used for the study. Migration assays were performed using transwell inserts. THP-1 cells were exposed to E2 or raloxifene in the presence of monocytic chemoattractant protein-1 (MCP-1), a major chemoattractant for monocytes. The cells were transfected with small interfering RNA (siRNA) against estrogen receptor (ER) alpha and ERbeta for gene silencing. ER expression was evaluated by Western blot analysis. RESULTS: MCP-1 induced the migration of the cells for 90 minutes. The addition of E2 or raloxifene significantly inhibited the MCP-1-induced migration for 90 minutes. Preincubation of THP-1 cells with an ER antagonist, ICI 182780, significantly attenuated the inhibitory effects of E2 and raloxifene. Whereas transfection with siRNA of ERalpha significantly attenuated the inhibition by E2 of MCP-1-induced monocyte migration, transfection with control siRNA or siRNA of ERbeta had no effect on the rapid inhibitory action of E2. Moreover, preincubation of THP-1 cells with a transcriptional inhibitor, actinomycin D, had no effect on the rapid inhibitory action of E2. CONCLUSIONS: Our findings suggest that both E2 and raloxifene inhibited the MCP-1-induced monocyte migration through nongenomic ERalpha. This result may explain one of the antiatherosclerotic effects of E2 and raloxifene on vasculature.

Atherosclerosis↗