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Dive into the research topics where Makoto Usami is active.

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Featured researches published by Makoto Usami.


Nutrition Research | 2008

Butyrate and trichostatin A attenuate nuclear factor κB activation and tumor necrosis factor α secretion and increase prostaglandin E2 secretion in human peripheral blood mononuclear cells

Makoto Usami; Kazunori Kishimoto; Atsushi Ohata; Makoto Miyoshi; Michiko Aoyama; Yuri Fueda; Joji Kotani

The effects of short-chain fatty acids (butyrate, propionate, and acetate) and trichostatin A (TSA), a typical histone deacetylase inhibitor, on tumor necrosis factor (TNF)-alpha secretion and nuclear factor kappaB (NF-kappaB) activation in peripheral blood mononuclear cells induced with lipopolysaccharide were evaluated in relation to prostaglandin E(2) (PGE(2)) secretion. Treatment of cells with butyrate; tributyrin, a prodrug of butyrate; propionate; acetate; and TSA down-regulated TNF-alpha secretion but all up-regulated PGE(2) secretion. Butyrate, propionate, and TSA inhibited NF-kappaB activation. The effects of the cyclooxygenase-nonspecific inhibitor, indomethacin; the cyclooxygenase-2 selective inhibitor, N-[2-(cyclohexyloxy)-4-nitro-phenyl] methanesulfonamide; and the general lipoxygenase inhibitor, nordihydroguaiaretic acid, varied in cells treated with each short-chain fatty acids. N-[2-(cyclohexyloxy)-4-nitro-phenyl] methanesulfonamide inhibited the effect of propionate on TNF-alpha secretion, and nordihydroguaiaretic acid inhibited that of acetate. The results showed that butyrate, propionate, and TSA inhibited TNF-alpha production via PGE(2) secretion and down-regulated NF-kappaB activation by lipopolysaccharide. These data suggest that the mechanism of butyrate and propionate action is through histone deacetylation and acetate through lipoxygenase activation in the regulation of proinflammatory responses in cells.


Nutrition | 2003

Effect of γ-linolenic acid or docosahexaenoic acid on tight junction permeability in intestinal monolayer cells and their mechanism by protein kinase C activation and/or eicosanoid formation

Makoto Usami; Takako Komurasaki; Aki Hanada; Kaori Kinoshita; Atsushi Ohata

OBJECTIVE Polyunsaturated fatty acids have been characterized as immunonutrients, but the effect of gamma-linolenic acid (GLA) or docosahexaenoic acid (DHA) on intestinal permeability has rarely been reported. METHODS Confluent Caco-2 cells on porous filter were used to measure tight junction function by fluorescein sulfonic acid permeability and transepithelial electrical resistance. Treatments with 0, 10, 50, and 100 microM of GLA or DHA during 24 h were compared. Then the effects of butylated hydroxytoluene (antioxidant), 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (protein kinase C antagonist), and inhibitors of enzymatic degradation to the eicosanoids, indomethacin (cyclooxygenase inhibitor) and 2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-p-benzoquinone (lipoxygenase inhibitor), on GLA or DHA were examined. RESULTS GLA and DHA enhanced fluorescein sulfonic acid permeability to 8.7- and 1.4-fold, respectively, and lowered transepithelial electrical resistance to 0.52- and 0.73-fold, respectively, versus the control in a concentration-dependent manner without cell injury (P < 0.001 to 0.05). Indomethacin and 2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-p-benzoquinone enhanced the changes mediated by GLA but did not alter the DHA effect. Butylated hydroxytoluene was ineffective. 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine facilitated the changes mediated by GLA, DHA, and eicosapentaenoic acid. The results indicated that the mechanism to change tight junction permeability via protein kinase C regulation is common but that via eicosanoid formation differs among GLA, DHA, and eicosapentaenoic acid. CONCLUSIONS GLA and DHA affect tight junction permeability in intestinal monolayer cells specifically and in a concentration-dependent manner.


Journal of Artificial Organs | 2003

Experimental proof of contamination of blood components by (1→3)-β-D-glucan caused by filtration with cellulose filters in the manufacturing process

Koichi Nagasawa; Tatsuyuki Yano; Go Kitabayashi; Hiroaki Morimoto; Yuji Yamada; Atsushi Ohata; Makoto Usami; Takashi Horiuchi

Abstract The level of (1→3)-β-D-glucan in blood is a diagnostic index of fungal infection because it is released from the fungal cell wall. However, high levels of plasma (1→3)-β-D-glucan in patients administered blood components may give false positive results. High levels of (1→3)-β-D-glucan have been detected in blood components. We suspected that (1→3)-β-D-glucan from cellulose filters had been eluted into blood components by filtration in the manufacturing process. To investigate the contamination of blood components by (1→3)-β-D-glucan from cellulose filters, in vitro experiments were performed by using six cellulose filters and a nylon filter. Human serum albumin (HSA) solution (100 ml) was flowed through each filter after rinsing with 100 ml of distilled water, and (1→3)-β-D-glucan in each fraction was determined by Fungitec G test MK. The concentration of (1→3)-β-D-glucan eluted from cellulose filters in 100-ml distilled water fractions ranged from 6 to 207 pg/ml, and that of HSA fractions ranged from 33 to 20,784 pg/ml. These data showed that remarkably higher (1→3)-β-D-glucan levels were detected in HSA fractions flowed through cellulose filters in spite of advance rinsing with 100 ml of distilled water. In the case of a nylon filter, (1→3)-β-D-glucan was not eluted in either fraction. These results indicate that (1→3)-β-D-glucan contamination in blood components is caused by filtration with cellulose filters in the manufacturing process.


Therapeutic Apheresis and Dialysis | 2014

Proteomic Analysis of Proteins Eliminated by Low‐Density Lipoprotein Apheresis

Yumiko Yuasa; Tsukasa Osaki; Hisashi Makino; Noriyuki Iwamoto; Ichiro Kishimoto; Makoto Usami; Naoto Minamino; Mariko Harada-Shiba

Low‐density lipoprotein apheresis (LDL‐A) treatment has been shown to decrease serum LDL cholesterol levels and prevent cardiovascular events in homozygous patients with familial hypercholesterolemia. Recently, LDL‐A treatment has been suggested to have beneficial effects beyond the removal of LDL particles. In this study, to clarify the preventive effects of LDL‐A treatment on atherosclerosis, the waste fluid from the adsorption columns was analyzed. The waste fluid of LDL adsorption columns was analyzed by two‐dimensional electrophoresis followed by mass spectrometry. Serum concentrations of the newly identified proteins before and after LDL‐A treatment were measured by enzyme‐linked immunosorbent assay. We identified 48 kinds of proteins in the waste fluid of LDL adsorption columns, including coagulation factors, thrombogenic factors, complement factors, inflammatory factors and adhesion molecules. In addition to the proteins that were reported to be removed by LDL‐A treatment, we newly identified several proteins that have some significant roles in the development of atherosclerosis, including vitronectin and apolipoprotein C‐III (Apo C‐III). The serum levels of vitronectin and Apo C‐III decreased by 82.4% and 54.8%, respectively, after a single LDL‐A treatment. While Apo C‐III was removed with very low‐density lipoprotein (VLDL) and LDL, vitronectin was removed without association with lipoproteins. The removal of proteins observed in the waste fluid has a certain impact on their serum levels, and this may be related to the efficacy of LDL‐A treatment. Proteomic analysis of the waste fluid of LDL adsorption columns may provide a rational means of assessing the effects of LDL‐A treatment.


Asaio Journal | 1989

Therapeutic plasmapheresis for liver failure after hepatectomy

Makoto Usami; Ohyanagi H; Nishimatsu S; Kasahara H; Shiroiwa H; Ishimoto S; Ueda T; Saitoh Y

Therapeutic plasmapheresis (PP) for liver failure following the resection of primary liver cancer was evaluated retrospectively. PP was repeated two to three times per week using plasma exchange (PE) with fresh frozen plasma as a substitution fluid in all eight cases and direct hemoperfusion (DHP) in two cases. PE effectively decreased serum bilirubin and/or improved hepatic encephalopathy in six of eight patients and prolonged survival time (p less than 0.05, chi-square test). DHP was not effective. Liver failure following the resection of damaged liver differs from acute liver failure in both the condition itself and the reaction to PP. Fractionation of serum bilirubin using high performance liquid chromatography showed differences in the ratio of delta bilirubin in one survivor versus two who died and was effective in selecting a patient who would respond to PE. In conclusion, PE is effective in prolonging survival time after liver failure in hepatectomized patients compared to conventional therapy.


Nutrition | 1996

The effect of a nucleotide-nucleoside solution on hepatic regeneration after partial hepatectomy in rats

Makoto Usami; Kouji Furuchi; Mitsutoshi Ogino; Hiroshi Kasahara; Taichi Kanamaru; Yoichi Saitoh; Hiroomi Yokoyama; Seiichiro Kano

After hepatectomy, purine and pyrimidine metabolism is a key process in the synthesis of DNA and RNA and maintaining cellular energy metabolism. The purpose of this study is to evaluate changes in blood purine and pyrimidine levels after partial hepatectomy and the effect of purine and pyrimidine nucleoside solution injection on hepatic regeneration under the hypothesis that the rat after partial hepatectomy requires substrates for salvage nucleotide synthesis and changes blood nucleoside and nucleobase levels. Blood levels of nucleotides, nucleosides, and nucleobase by high-performance liquid chromatography method and liver ATP level by enzymatic analysis, and the effect of preoperative injection of nucleoside solution (OG-VI) on hepatic regeneration ratio and hepatocytes DNA synthesis, were assessed in rats after 70% partial hepatectomy. Decreased liver adenosine triphosphate and increased plasma xanthine and hypoxanthine after partial hepatectomy indicated an increase in catabolism of purine nucleotides in regenerating liver. Plasma thymidine and cytidine levels increased, then returned to the prevalue, suggesting that the thymidine and cytidine pool was enlarged. OG-VI increased labeling indices of hepatocytes at postoperative d 1 (POD) and hepatic regeneration ratio at POD 14. Blood purine nucleobase and pyrimidine nucleoside levels change after partial hepatectomy and preoperative supply of nucleoside solution is effective for increasing hepatocytes DNA synthesis and hepatic regeneration after partial hepatectomy.


Nutrition | 1996

Effect of a parenteral nucleoside-nucleotide mixture on hepatic metabolism in partially hepatectomized cirrhotic rats

Makoto Usami; Atsunori Iso; Hiroshi Kasahara; George Kotani; Seiji Haji; Taichi Kanamaru; Yoichi Saitoh

After hepatectomy, purine and pyrimidine metabolism is a key process in synthesis of DNA and RNA and in energy metabolism. To supply nucleosides for salvage synthesis, nucleoside-nucleotide mixture solutions have been developed, and they have been found to improve protein metabolism and hepatic regeneration after partial hepatectomy in normal rats. However, the effect of the solution in cirrhotic liver, common in patients with hepatocellular carcinoma, has not been reported. The aim of this study was to evaluate the metabolic effect of the nucleoside-nucleotide mixture on cirrhotic rats after partial hepatectomy. Seventy percent partial hepatectomy was performed in thioacetamide-administered cirrhotic rats. The fractional protein synthetic rate, nitrogen balance, hepatic content of nucleic acid, and blood chemistry after the administration of the nucleoside-nucleotide mixture solution (OG-VI) with total parenteral nutrition was evaluated at 7 d after partial hepatectomy. OG-VI increased hepatic RNA level and hepatic fractional protein synthetic rate (p < 0.05). It is concluded that the nucleoside-nucleotide mixture solution is an effective nutritional supplement to the metabolism of cirrhotic rats after partial hepatectomy.


Surgical Infections | 2014

Intravenous Immunoglobulin-Induced Neutrophil Apoptosis in the Lung during Murine Endotoxemia

Ayumi Seishu; Saori Kawakami; Noriaki Maeshige; Makoto Miyoshi; Takahiro Ueda; Makoto Usami; Atsunori Nakao; Joji Kotani

BACKGROUND The pathophysiologic features of acute respiratory distress syndrome (ARDS) are attributed to neutrophil accumulation and over-activation. Low blood immunoglobulin G concentrations in septic shock patients are associated with higher risk of developing ARDS. This study showed the effects of intravenous immunoglobulin (IVIg) on neutrophil apoptosis and accumulation in the lung during murine endotoxemia. METHODS Male C57BL/6J mice were injected with saline or 7 mg/kg of lipopolysaccharide (LPS), and 3 h later also were injected with saline, IVIg 300 mg/kg, or IVIg 1000 mg/kg intraperitoneally. At 12 h after LPS injection, mice were sacrificed and peripheral blood and lungs were collected. The lung messenger ribonucleic acid expression (tumor necrosis factor-α [TNF-α], inducible nitric oxide synthase [iNOS], and intercellular adhesion molecule-1 [ICAM-1]) was determined using quantitative realtime reverse transcriptase-polymerase chain reaction. Lungs were immersed in 4% paraformaldehyde and then embedded in paraffin. Tissue slices were prepared and stained with naphthol AS-D chloroacetate esterase to detect neutrophils. The numbers of neutrophils (characterized by the segment number of their nuclei) were counted. Peripheral neutrophil apoptosis was detected by annexin V using flow cytometry and lung neutrophil apoptosis was detected by cleaved caspase-3 using immunohistochemistry. RESULTS The survival rates of the saline group, LPS group, and IVIg group were all 100%. Apoptosis of peripheral blood neutrophils was inhibited by LPS. Neutrophil accumulation in the lung was decreased by both IVIg 300 mg/kg and 1000 mg/kg. Segmented neutrophils were reduced by IVIg during endotoxemia. However, IVIg 300 mg/kg and 1000 mg/kg had no influence on the lung messenger ribonucleic acid expression of TNF-α, iNOS, or ICAM-1. Cleaved-caspase-3-positive neutrophils were increased in the IVIg 300 mg/kg group during endotoxemia. The 1000 mg/kg IVIG dose reduced the number of segmented neutrophils, but did not induce cleaved-caspase 3-positive neutrophils. CONCLUSION A therapeutic IVIg dose can attenuate neutrophil accumulation and regulate neutrophil apoptosis in the lung during endotoxemia. It is possible that the pathways by which IVIG induces neutrophil apoptosis may differ depending on the IVIg concentration.


Nutrition | 1997

The effect of a nucleotide-nucleosidesolution on hepatic regeneration in rats after partial hepatectomy and in primary monolayer culture of hepatocytes

Makoto Usami; Yoichi Saitoh

Purine and pyrimidine metabolism is a key process after hepatic surgery. To evaluate the effect of purine and pyrimidine supplementation on hepatic regeneration, the following clinical in vitro and in vivo experiments were performed. Changes in blood nucleotides, nucleosides and nucleobase were analyzed by high performance liquid chromatography in patients and rats after partial hepatectomy. The effect of supplementation of nucleotide-nucleoside solution (OG-VI) or their components on nucleic acids syntheses in primary monolayer cultured hepatocytes and preoperative intraperitoneal injection of OG-VI on hepatic regeneration in the partially hepatectomized rats were evaluated. Blood purine and pyrimidine levels in patients change after hepatectomy and their changes indicate increased salvage synthesis of purine and pyrimidine in the regenerating liver. Addition of appropriate amounts of inosine, GMP, uridine, or thymidine, the substrates for salvage purine and pyrimidine syntheses, to primary cultures of hepatocytes enhanced both DNA and RNA syntheses by the salvage and de novo pathways. The OG-VI mixture also enhanced the syntheses of DNA and RNA. Preoperative administration of OG-VI to partially-hepatectomized rats enhances hepatic DNA synthesis in a way similar to the in vitro study.


Advances in Experimental Medicine and Biology | 1995

The effect of partial hepatectomy on blood purine levels in rats and patients.

Makoto Usami; Furuchi K; Hiroshi Kasahara; Seiji Haji; Kitani G; Atsunori Iso; Kai Sun; Sou E; Zheng Jh; Kazuya Sakata

Purine metabolism is a key metabolic process after partial hepatectomy in the synthesis of DNA and RNA, and in the energy metabolism for hepatic regeneration1. However it is not evaluated in clinical situations due to the difficulty in measuring changes of purine levels in the liver itself. In order to get a basic understanding of purine metabolism in clinical hepatectomy cases, blood samples are used for analysis in this study. Also, the liver is the main nondietary source of purines in mammalian cells2 and changes in blood level may affect purine metabolism in various tissues.

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