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ABSTRACT: Malignant pleural mesothelioma (MPM) is an aggressive malignancy for which there is no approved targeted therapy. We examined the therapeutic efficacy of the mitogen-activated protein kinase kinase (MEK) and phosphatidylinositol 3-kinase (PI3K) inhibitors against human MPM cell lines both in vitro and orthotopically inoculated into severe combined immunodeficient (SCID) mice. In addition, the molecular mechanisms of these agents were confirmed in vitro and in vivo. The MEK or the PI3K inhibitor suppressed MPM cell growth in vitro in a dose-dependent manner via induction of G1 cell cycle arrest and apoptosis. In addition, combined use of the MEK and PI3K inhibitors showed an additive or synergistic inhibitory effect on MPM cell growth compared to treatment with either individual drug. Treatment with MEK or PI3K inhibitor suppressed the production of thoracic tumors and pleural effusion and prolonged the survival time of EHMES-10 cell-bearing SCID mice. The combination therapy more effectively prolonged the survival time compared to treatment with either individual drug. Immunohistochemical and western blot analysis of thoracic tumors suggested that these agents induced cell cycle arrest, apoptosis and inhibition of tumor angiogenesis. Our results suggest that a combination of MEK and PI3K inhibitors is a promising therapeutic strategy for MPM.
International Journal of Oncology 05/2012; 41(2):449-56. · 2.40 Impact Factor
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Kazunori Irifune,
Hironobu Hamada,
Ryoji Ito,
Hitoshi Katayama,
Akira Watanabe, Aki Kato,
Seigo Miyoshi,
Naohiko Hamaguchi,
Ryo Toyozawa,
Sachiko Hamaguchi,
Masahiro Abe,
Kazutaka Nishimura,
Jitsuo Higaki
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ABSTRACT: Bakumondoto (TJ-29) is a traditional herbal medicine that has been used in Japan for the treatment of bronchitis, bronchial asthma, and cough. This study investigated the effect of TJ-29 for the treatment of post-infectious prolonged cough. We performed a multicenter randomized controlled trial treating patients without (group A, n=11) or with TJ-29 (group B, n=8) for a total of 2 weeks using a beta 2 stimulant as the basal agent. Efficacy and safety were compared by a cough diary, VAS and sleeping questionnaire. At 4 and 5 days after treatment, the cough score of group B showed significant improvement compared with group A, demonstrating an early antitussive effect. At the assessment 2 weeks after treatment start, both groups showed similar levels of improvement in the cough score. No significant difference was observed in the VAS and the sleeping questionnaire items. In conclusion, oral TJ-29 administration could be useful and safe for the treatment of post-infectious prolonged cough.
Phytomedicine: international journal of phytotherapy and phytopharmacology 06/2011; 18(8-9):630-3. · 2.17 Impact Factor
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ABSTRACT: The pathology of diabetic retinopathy involves endothelial dysfunction, in which leukocyte adhesion to the vascular endothelium via intercellular adhesion molecule-1 (ICAM-1) may play a key role. Short-interfering RNAs (siRNAs) are unique modulators of gene expression in mammalian cells. The purpose of this study was to evaluate the enhanced effect of hyperglycemia and the attenuating effect of siRNAs on ICAM-1 expression in cultured endothelial cells.
Human umbilical vein endothelial cells (HUVECs) were seeded onto 24-well culture plates. The following day, ICAM-1-specific siRNAs were transfected using Lipofectamine 2000. Glucose (15, 30, or 45 mM) or interleukin-1ss as a positive control was added to the medium to stimulate ICAM-1. After 48 hours, the HUVECs were collected to measure the ICAM-1 expression by enzyme-linked immunosolvent assay. Fluoresceinated siRNAs were transfected into HUVECs to confirm transfection of the siRNAs into HUVECs by fluorescein microscopy.
Glucose enhanced the ICAM-1 expression in a dose-dependent manner. ICAM-1 expression stimulated by hyperglycemia decreased significantly in the HUVECs transfected with corresponding siRNAs. Transfection of siRNAs was confirmed with enhanced fluorescence in HUVECs incubated with control siRNAs.
These results suggested that hyperglycemia stimulated protein expression of ICAM-1 and that siRNAs suppressed gene expression of ICAM-1 in HUVECs. The RNA-targeting approach using siRNAs may provide a novel therapeutic option for diabetic retinopathy.
Albrecht von Graæes Archiv für Ophthalmologie 08/2008; 246(7):989-92. · 2.17 Impact Factor
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ABSTRACT: Mannose is an unusable carbon source for many plants. In our study we compared the effects of mannose and sucrose on growth and sucrose levels in azuki bean (Vigna angularis) cells grown in liquid media and in solid media. The suspension cells grew actively in a liquid medium containing 90 mM sucrose but not in that containing 90 mM mannose, where the intracellular sucrose levels were reduced to 20% or less of those in sucrose-grown cells. These results suggested that the limited conversion of mannose to sucrose resulted in cell growth inhibition. When sucrose-grown suspension cells (1 x 10(5)) were transferred onto agar medium containing mannose, they grew little initially, but, after a month lag period, they started to form many callus colonies at a high apparent variation rate (1.3 x 10(-3)). Time-course studies for sugar and enzyme analysis revealed that the mannose-accommodated cells were capable of converting mannose to sucrose, with enhanced phosphomannose isomerase activity. The mannose-accommodated cells actively grew in liquid medium with sucrose but lost their ability to grow with mannose again, suggesting a specific trait of callus culture for mannose utilization. The possible differences in the metabolic activities and other physiological characteristics are discussed between callus and suspension cells.
Journal of Plant Research 06/2008; 121(3):339-49. · 1.75 Impact Factor
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ABSTRACT: Plant cells utilize various sugars as carbon sources for growth, respiration and biosynthesis of cellular components. Suspension-cultured cells of azuki bean (Vigna angularis) proliferated actively in liquid growth medium containing 1% (w/v) sucrose, glucose, fructose, arabinose or xylose, but did not proliferate in medium containing galactose or mannose. These two latter sugars thus appeared distinct from other sugars used as growth substrates. Galactose strongly inhibited cell growth even in the presence of sucrose but mannose did not, suggesting a substantial difference in their effects on cell metabolism. Analysis of intracellular soluble-sugar fractions revealed that galactose, but not mannose, caused a conspicuous decrease in the cellular level of sucrose with no apparent effects on the levels of glucose or fructose. Such a galactose-specific decrease in sucrose levels also occurred in cells that had been cultured together with glucose in place of sucrose, suggesting that galactose inhibits the biosynthesis, rather than uptake, of sucrose in the cells. By contrast, mannose seemed to be metabolically inert in the presence of sucrose. From these results, we conclude that sucrose metabolism is important for the heterotrophic growth of cells in plant suspension-cultures.
Journal of Plant Research 12/2007; 120(6):713-9. · 1.75 Impact Factor
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ABSTRACT: An 83-year-old man was admitted with paraplegia and loss of all sensation below the level of umbilicus, with bowel and bladder dysfunction. Stage IV small cell lung cancer had been diagnosed two years ago and had received several courses of chemotherapies. A magnetic resonance imaging revealed an enhanced mass in the intramedullary spinal cord at the level of Th10-L1. Metastatic spinal tumor was diagnosed by clinical and radiological examinations. This is a rare case of small cell lung cancer with intramedullary spinal cord metastasis which caused various neurological symptoms.
Nippon Ronen Igakkai Zasshi Japanese Journal of Geriatrics 10/2005; 42(5):567-70.
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ABSTRACT: To evaluate the inhibitory effect of triamcinolone acetonide (TA) on choroidal neovascularization (CNV) by posterior sub-Tenon administration using a laser-induced CNV model in the rat.
Experimental CNV was induced by laser photocoagulation in Brown-Norway male rats. Experimental eyes received posterior sub-Tenon administration of either 2 mg (n = 10) or 0.5 mg (n = 8) of TA. Control eyes (n = 10) received posterior sub-Tenon administration of isotonic sodium chloride solution. Two weeks after treatment, CNV was evaluated by fluorescein angiography and histopathological examination. Concentrations of TA in the vitreous, retina, and choroid were determined by high-performance liquid chromatography at 3 and 7 days after posterior sub-Tenon administration.
The eyes treated with 2 mg of TA showed statistically significant inhibition of fluorescein leakage by fluorescein angiography, as compared with control eyes and eyes treated with 0.5 mg of TA (P < 0.01). The thickness of CNV membranes in eyes treated with 2 mg of TA also decreased statistically significantly, as compared with control eyes (P < 0.01). TA was detected in the vitreous, retina, and choroid 3 days after administration and in the choroid 7 days after administration.
Posterior sub-Tenon administration of TA may be useful to treat CNV.
Retina 06/2005; 25(4):503-9. · 2.81 Impact Factor
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ABSTRACT: To investigate the effect and safety of benzalkonium chloride on transscleral drug delivery in the rabbit after continuous intrascleral administration.
Betamethasone 21-phosphate (BP) aqueous solutions, with or without benzalkonium chloride (BAK), were continuously administered to albino rabbit sclera with an osmotic pump for 1 week. The BP concentrations in the aqueous humor, vitreous, and retina-choroid were measured by high-performance liquid chromatography (HPLC). To investigate the effect of BAK on scleral permeability of BP in vitro, penetration of BP aqueous solution with or without BAK across the rabbit sclera was evaluated using a two-chamber Ussing apparatus. To determine the effects of BAK on transscleral delivery of large molecules, 20- and 70-kDa fluorescein isothiocyanate (FITC)-dextran (FD-20 and -70, respectively) aqueous solutions, with or without BAK, were continuously administered to the sclera by an osmotic pump. The intensity of fluorescence in the aqueous humor, vitreous, and retina-choroid was measured by fluorescence spectrophotometry at 1 week after implantation of the pump. The retinal toxicity of BAK was evaluated electrophysiologically and histologically.
BAK increased concentrations of BP in the vitreous and retina-choroid compared with the control. BP was not detected in the aqueous humor. In the in vitro study, BAK did not increase the scleral permeability of BP. In the retina-choroid, BAK significantly increased concentrations of FD-20 but did not increase those of FD-70. The addition of BAK did not increase concentrations of FD-20 or -70 in the vitreous. No substantial toxic reactions were observed in the retina in electrophysiological or histologic examinations after the addition of BAK.
The results of this study demonstrate that BAK may improve the ocular penetration of a drug in a transscleral drug delivery system without producing toxic reactions.
Investigative Ophthalmology & Visual Science 03/2005; 46(2):703-8. · 3.60 Impact Factor
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ABSTRACT: To evaluate the feasibility of a nonbiodegradable polymeric episcleral implant as a new controlled intraocular delivery system of betamethasone (BM) to the posterior pole of the eye.
The episcleral implant, which is composed of a drug-releasing component and a suture tag, released BM through an ethylene vinyl acetate membrane. The implants were placed on the sclera in 12 eyes of 12 Japanese white rabbits so that the drug-releasing surface could attach to the sclera at the posterior pole. BM concentrations in the aqueous humor, vitreous, and retina-choroid (posterior half and anterior half) were determined by high-performance liquid chromatography (HPLC) at weeks 1, 2, and 4 after implantation. In addition, the intraocular tissue distribution of the drug was evaluated by fluorescein microscopy after implantation of the implant loaded with 6-carboxy fluorescein diacetate (6-CFDA) as a drug marker. Retinal toxicity was evaluated by electroretinography and histologic examination.
The implant showed zero-order release profiles both in vitro and in vivo for 4 weeks. BM concentrations in the retina-choroid after implantation were maintained above the concentrations effective for suppressing inflammatory reactions for at least 4 weeks. The BM concentration was greater in the posterior half of the retina-choroid than in the vitreous. It was confirmed that 6-CFDA penetrated through the sclera and dispersed into the retina-choroid. Fluorescence from 6-CFDA gradually decreased in intensity with increased distance from the implantation site. Electroretinography and histologic study showed no substantial toxic reactions.
These findings suggest that the episcleral implant may be a useful drug carrier for intraocular delivery of BM, especially for the posterior part of the eye.
Investigative Ophthalmology & Visual Science 02/2004; 45(1):238-44. · 3.60 Impact Factor
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ABSTRACT: To evaluate the tissue distribution of betamethasone (BM) after implantation of a nonbiodegradable intrascleral implant as a new, controlled intraocular delivery system.
Nonbiodegradable intrascleral implants designed to release BM for at least 1 month were placed in the sclera of pigmented rabbits. The BM concentrations in the aqueous humor, vitreous, and retina-choroid were determined by high-performance liquid chromatography (HPLC) at 3, 7, 14, and 28 days after implantation. The BM concentrations in three sections of retina-choroid were also investigated. Retinal toxicity was evaluated by electroretinography and histology.
The BM released from the intrascleral implant in vitro and in vivo showed zero-ordered release profiles for 4 weeks. The BM concentrations in the retina-choroid after placement of the intrascleral implants remained higher than effective concentrations for suppressing various inflammatory processes for at least 28 days. The BM concentrations in the retina-choroid around the implantation site were more than 10 times higher than in the opposite side throughout the study. No substantial toxic reactions were observed by electroretinography or histology.
These findings suggested that the nonbiodegradable intrascleral implant could be a useful drug carrier for intraocular delivery of BM without producing severe retinal toxicity. The intrascleral site may be considered for effective intraocular drug distribution after implantation.
Investigative Ophthalmology & Visual Science 07/2003; 44(6):2702-7. · 3.60 Impact Factor
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ABSTRACT: To evaluate the feasibility of using a biodegradable intrascleral implant for intraocular sustained delivery of betamethasone phosphate (BP).
The intrascleral implant (0.5 mm thick and 4 mm in diameter) was made of poly(DL-lactide) containing 25% betamethasone phosphate. The in vitro release of BP from the implant was evaluated by high-performance liquid chromatography (HPLC). The implants were placed into a scleral pocket in the rabbit's eye. The concentrations of BP in the aqueous humor, vitreous, and retina-choroid were measured by HPLC. The toxicity and biocompatibility of the implant were evaluated by slit lamp examination, electroretinography, and light microscopy.
In vitro studies demonstrated that the implants released BP in a biphasic pattern for at least 8 weeks. The BP concentrations in the vitreous and the retina-choroid remained within the concentration range capable of suppressing inflammatory responses for more than 8 weeks. The BP concentration was greater in the retina-choroid than in the vitreous. In the aqueous humor, BP was below the detection limit during the observation period. No significant toxicity to the retina was observed. Also, the implant showed good biocompatibility in the eye.
These results suggest that the intrascleral implant would be a promising system for delivery of steroid to the posterior segment of the eye.
Investigative Ophthalmology & Visual Science 03/2003; 44(2):740-4. · 3.60 Impact Factor
