Atsushi Ohata

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

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.

Positive (1→3)‐β‐d‐glucan in blood components and release of (1→3)‐β‐d‐glucan from depth‐type membrane filters for blood processing

BACKGROUND : The false‐positive elevation of plasma (1→3)‐β‐ d ‐glucan level, a serodiagnostic test for deep‐seated mycosis, is suspected in patients administered with blood components.

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

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.

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

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.

Phospholipid Fatty Acid Composition and Diamine Oxidase Activity of Intestinal Mucosa From Rats Treated With Irinotecan Hydrochloride (CPT-11) under Vegetable Oil–Enriched Diets: Comparison Between Perilla Oil and Corn Oil

BACKGROUND Irinotecan hydrochloride (CPT-11), a topoisomerase I inhibitor highly effective for various cancers, has its dosage limited by diffuse mucosal damage with increased prostaglandin (PG) E(2). However, an analysis of intestinal phospholipid fatty acid composition after CPT-11 treatment has not been reported. This study aimed to evaluate intestinal phospholipid fatty acid composition in relation to intestinal mucosal integrity and plasma and mucosal PGE(2) levels after CPT-11 treatment. The effect of dietary vegetable oil supplementation, perilla oil vs corn oil, was also evaluated. METHODS Intestinal phospholipid fatty acid composition, PGE(2) level, mucosal diamine oxidase (DAO) activity, diarrhea, and blood tests were evaluated in rats injected with CPT-11 under a conventional diet. The same parameters were compared among 3 different dietary vegetable oil supplementations: perilla oil, corn oil, and a 1:3, respectively, mixture with a semisynthetic diet during 14 days. RESULTS CPT-11 treatment caused severe diarrhea, and intestinal mucosal fatty acid composition changed with increased PGE(2) level and decreased DAO activity. Decreases in eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and EPA/arachidonic acid (AA) ratio in colonic mucosa were observed. Perilla oil increased omega-3 polyunsaturated fatty acids, alpha-linolenic acid, EPA, and EPA/AA ratio and decreased plasma PGE(2). But the amounts used were not enough to attenuate intestinal damage from CPT-11 treatment. CONCLUSIONS CPT-11 induced changes of intestinal mucosal fatty acid composition with increased PGE(2) level and decreased intestinal integrity; perilla oil shows the possibility of being able to attenuate those changes.