Dietary phosphatidylinositol protects C57BL/6 mice from concanavalin A-induced liver injury by modulating immune cell functions.

SCOPE: Several recent studies have demonstrated that phospholipids (PLs) supplementation can modulate the function of cultured-immune cells. Furthermore, dietary PLs have been shown to ameliorate inflammatory processes and immune responses in arthritic and diabetic murine models, respectively. Thus, the aim of this study was to examine the immune-modulating activities of dietary soybean PLs in mice, with particular emphasis on the immune cell functions. METHODS AND RESULTS: Mice were fed semisynthetic diets for 6 weeks, which contained either 7% soybean oil or 5% soybean oil plus 2% of either PL: phosphatidylcholine (PC), phosphatidylinositol (PI), or phosphatidylserine (PS). Production of concanavalin A (Con A)-induced proinflammatory cytokines was significantly decreased in the splenocytes isolated from mice fed PI compared to other lipids. Supplementation of the diet with PI, but not with the other lipids, significantly suppressed the proinflammatory cytokine serum levels and the development of Con A-induced liver damages. CONCLUSION: These observations suggest that dietary PI influenced immune functions, resulting in the prevention of pathogenesis and development of the liver injury in mice.

[A rare case report of incarceration of Lambl's excrescence of aortic valve resulting in myocardial infarction].

We report a rare autopsy case of Lambl's excrescence of the aortic valve resulting in myocardial infarction. A 50-year-old female patient was referred to our hospital, but she died 4 hours after admission due to myocardial infarction. Autopsy disclosed Lambl's excrescence of the aortic valve, which obstructed the main branch of the left coronary artery. Histologically, fibrin thrombus and bacterial flora were found on surface of Lambl's excrescence. Differential diagnosis of infected Lambl's excrescence, so called endocarditis with infected vegetation, and papillary fibroelastomais discussed with a literature review.

Effect of Peucedanum japonicum Thunb extract on high-fat diet-induced obesity and gene expression in mice.

Supplementation of the diet with Peucedanum japonicum Thunb (PJT) powder inhibits high-fat diet-induced obesity in mice. Either the fiber component or other bioactive components in the PJT powder may inhibit obesity. This study, therefore, was an attempt to identify the components, fiber or other phytochemicals of PJT that were responsible for the anti-obesity activity, and also studied the modulation of obesity-related gene expression in C57BL/6 mice. Animals were fed a modified-AIN76 diet supplemented with PJT powder or extracts of PJT in water, 50 % ethanol, or ethanol. Body weight gain, tissue weight, serum biochemical parameters, liver lipid concentrations, and gene expression in tissues were compared between the control and treatment groups. Of the extracts, the ethanol extract of PJT decreased fat accumulation and adipocyte size, reduced serum and liver triglyceride concentrations, and inhibited obesity. This finding clearly demonstrates the presence of anti-obesity phytochemicals in PJT. Ethanol extract of PJT inhibited lipase activity in vitro. Modulation of gene expression by PJT ethanol extract was largely similar to that by PJT powder in the hepatic and adipose tissues: RORC and PBEF1 were upregulated and DUSP1, INSIG2, and SERPINA12 were downregulated in the liver; FXRα and PPARγ were upregulated and PEG1/MEST, the size-marker of adipocytes, was downregulated in the adipose tissue. Furthermore, PJT ethanol extract increased the expression of the UCP3 gene in muscle. These results suggest that the anti-obesity phytochemicals in PJT lower lipid levels by inhibiting fat absorption and by modulating obesity-related gene expression in the liver, adipose tissue, and muscle.

Protective effects of fractional extracts from Panellus serotinus on non-alcoholic fatty liver disease in obese, diabetic db/db mice.

Non-alcoholic fatty liver disease (NAFLD) is emerging as the most common liver disease in industrialised countries. Various mushrooms have been used in Eastern folk medicine for the treatment of lifestyle diseases. We previously found that the dietary intake of powdered whole Panellus serotinus (Mukitake) alleviates NAFLD in obese, diabetic db/db mice. In the present study, we investigated the influence of Mukitake fractional extracts on the development of NAFLD in db/db mice. A significant reduction in the hepatic TAG content, macrovesicular hepatocytes and activities of key enzymes for de novo synthesis of the fatty acid was observed in both the water-soluble Mukitake extract (WE) diet and the ethanol-soluble Mukitake extract (EE) diet groups compared with the control diet group of the db/db mice. The serum level of monocyte chemoattractant protein-1 (MCP-1), which is known to exacerbate insulin resistance, was significantly decreased in the WE group. On the other hand, the serum level of adiponectin, which plays a protective role against the metabolic syndrome, was significantly increased in the EE group. Additionally, differential analysis between Mukitake and Shiitake, mycelia from the same family, using liquid chromatography time-of-flight MS technology revealed that only seven and five compounds exist in WE and EE from Mukitake, respectively. In conclusion, the present study demonstrated that Mukitake displays at least two different physiological actions that alleviate NAFLD: one through the reduction in inflammatory damage by its suppression in MCP-1 production and the other through an increase in level of serum adiponectin and the prevention of visceral fat accumulation.

Effect of Peucedanum japonicum Thunb on the expression of obesity-related genes in mice on a high-fat diet.

The present study describes the effect of Peucedanum japonicum Thunb (PJT) intake on the expression of obesity-related genes in mice fed a high-fat diet. To explore the mechanism underlying the effect of PJT, This study focused on the expression of genes, especially those related to obesity and metabolism syndrome, in the liver and adipose tissues. In agreement with our previous observations, intake of 10 % PJT for 4 weeks significantly reduced serum triglyceride (TG), leptin, abdominal fat, and adipocyte size. PJT also significantly increased fecal excretion of TG, decreased that of bile acid, and tended to increase the fecal excretion of total cholesterol. Microarray analysis was used to monitor changes in 324 metabolic syndrome-related genes in the liver. Statistically significant upregulation of PPP1R10, RORC, and PBEF1 genes and downregulation of DUSP1, INSIG2, and SERPINA12 genes were noted and confirmed by real-time RT-PCR. These changes were indicative of increased fatty acid oxidation in the maintenance of lipid homeostasis and insulin sensitivity in the livers of PJT-fed mice. PJT increased the expression of PPARγ, FXRα, DGAT1, and ATGL genes, suggesting an enhancement of adipocyte differentiation and normalization of functionality of adipose tissue.

Peucedanum japonicum Thunb inhibits high-fat diet induced obesity in mice.

This study examined the antiobese activity of Peucedanum japonicum Thunb (PJT) in mice. In the first experiment, 4-week-old C57BL/6 mice were fed seven different diets containing 15% corn oil and 0-20% PJT powder for 4 weeks. Feeding the 10% and 20% PJT diet suppressed the body weight gain and the accumulation of abdominal and subcutaneous fats. PJT reduced serum and liver levels of triglyceride and serum levels of leptin in a dose-dependent manner. PJT intake decreased the proportion of saturated fatty acids and increased polyunsaturated and n-3 fatty acids in the liver. To obtain more insight into the antiobese activity of PJT, its effect on lipid absorption and enzyme activities related to lipid metabolism was studied in the second experiment. There was an increased faecal excretion of triglyceride in mice fed 5% and 10% PJT diets. Fatty acid synthase activity was decreased while carnitine palmitoyltransferase activity was increased by 10% PJT intake. These findings pointed to the usefulness of PJT for the development of a safe natural agent to reduce obesity or body weight for the first time. The rationale for the lipid lowering mechanism of PJT and the candidate compound responsible for the observations have also been discussed.

Comparative study of the effect of basal diet formulation, dietary fat and cholesterol levels on the development of aortic atherosclerotic lesions in B6.KOR-Apoeshl mice.

To optimize the adequate atherogenic diet composition for nutritional atherosclerosis studies utilizing B6.KOR-Apoe(shl) mice, we investigated the effect of dietary cholesterol, AIN formula, and dietary fats on the development of atherosclerosis. Cholesterol supplementation (0.15-2%) for 12 weeks resulted in a dose-dependent increase in the development of atherosclerosis. Furthermore, there were no significant differences in the degree of atherosclerosis between B6.KOR-Apoe(shl) mice fed a modified-AIN-76 diet and those fed a modified-AIN-93M diet containing corn oil or soybean oil for 10 and 12 weeks. The present experiment indicates that the adequate dietary level of cholesterol was 0.15%, and that further studies are necessary to determine the optimal level of various types of dietary oils for nutritional atherosclerosis experiments in B6.KOR-Apoe(shl) mice.

Contamination of rice by etofenprox, diethyl phthalate and alkylphenols: effects on first delivery and sperm count in mice.

We observed that first delivery was delayed when a group of paired mice fed with non-organic common rice compared to a group fed with organic rice. This led us to hypothesize that pesticides or other soil contaminants may be responsible for the effect on mice reproduction. We then found that the non-organic rice was contaminated with a pesticide etofenprox and nonylphenol, butylphenol and diethyl phthalate which are used as agricultural detergents or plasticizers of agricultural film, that are all suspected to be estrogenic. Therefore, the chemicals were administered to mice at the levels detected in rice, and we subsequently observed that first delivery and sperm count of the animals were significantly impaired. This study is the first to show that rice contaminated with agricultural chemicals affects reproduction in the mammal.

[Rare case report of solitary ganglioneuroma of the transverse colon].

We report herein a rare case of solitary ganglioneuroma occurring in the transverse colon with a brief literature review. A 45-year-old man was diagnosed as having hemorrhoids by a local medical practitioner and referred to our hospital for further examination. He showed neither signs nor symptoms of neurofibromatosis and multiple endocrine neoplasia. Colonic endoscopic examination demonstrated that a pedunculated polyp with a size of 11 mm in the diameter in the transverse colon. Histopathologic and immunohistochemical examination demonstrated that the endoscopic mucosal resection specimen of the polyp had abundant ganglionic cells, Schwann cells, and nerve fibers in the mucosa and submucosa.

Tumor-selective cytotoxicity of benzo[c]phenanthridine derivatives from Toddalia asiatica Lam.

PURPOSE: To develop a novel anti-cancer drug of low side effect against lung adenocarcinoma, the authors screened the bioresources of Okinawa Island, Japan. The medicinal plant Toddalia asiatica Lam. contained three benzo[c]phenanthridine derivatives: dihydronitidine (DHN), nitidine (NTD) and demethylnitidine (DMN). Of the three derivatives, DHN had been shown to selectively inhibit the growth of cancer cells in our previous study. Because of similar molecular topology of NTD or DMN to DHN, it can be expected that NTD and DMN also show selective cytotoxicity. The aim of the present study was therefore to examine the selective cytotoxicity of these two compounds in vitro and in vivo. METHODS: Benzo[c]phenanthridine derivatives were isolated from T. asiatica Lam., and their chemical structures were identified by interpretation of NMR and MS spectrum. Of the isolated compounds, NTD and DMN were evaluated for cytotoxicity in vitro or in vivo. RESULTS: NTD as well as DHN selectively reduced the growth of murine and human lung adenocarcinoma in vitro with selective intracellular accumulation. NTD has also been proven to be highly effective in vivo to inhibit the growth of both murine and human lung adenocarcinoma in a subcutaneous xenograft model without any deteriorating side effect. In contrast, DMN had no selective cytotoxicity suggesting that 8-methoxy group of NTD is the critical structural feature for the selective cytotoxicity. CONCLUSIONS: This study thus proves the effectiveness of benzo[c]phenanthridine derivatives as anti-cancer agent in vivo for the first time, and discusses the mechanisms responsible for the selective cytotoxicity.

Antiatherosclerotic function of Kokuto, Okinawan noncentrifugal cane sugar.

In the present study, we investigated the effect of phenolic compounds (PCs) and policosanol of Kokuto, Okinawan noncentrifugal cane sugar, on the development of atherosclerosis. A total of 67 male Japanese quail were divided into eight dietary groups in trial 1. The dietary groups were fed the atherosclerotic diet (AD) containing 5% corn oil, 2% cholesterol, and 30% sucrose or seven different types of Kokuto. Dietary intakes of Kokuto notably prevented the development of atherosclerosis. Furthermore, there was a significant negative correlation between the serum radical scavenging activity and the degree of atherosclerosis in the dietary groups. In trial 2, a total of 63 Japanese quail were fed AD with sucrose, Kokuto, PC extracts from Kokuto, wax extracts from sugar cane, octacosanol, vitamin C, and vitamin E. As a result, the supplementation of the diet with Kokuto and PCs significantly reduced the development of atherosclerosis as compared with the ingestion of AD with sucrose. In conclusion, these findings suggest that, among various components of Kokuto, PCs play a central role for the prevention of experimental atherosclerosis in Japanese quail.

Tumor specific cytotoxicity of beta-glucosylceramide: structure-cytotoxicity relationship and anti-tumor activity in vivo.

This study describes the structure-cytotoxicity relationship of beta-glucosylceramide (beta-GluCer) and its antitumor activity in vivo. Unglycosylated ceramide had no selective cytotoxicity which demonstrated that the sugar moiety plays a critical role for the expression of selective cytotoxicity by beta-GluCer. beta-Galactosylceramide also showed tumor specific cytotoxicity suggesting that the chemical structure of sugar group is not a factor for the selective toxicity. Similarly, unglycosylated ceramides of short acyl chain also selectively inhibited the growth of cancer cells. These findings in concert point to the importance of the hydrophilicity of the ceramide molecule rather than the chemical structure for the cyto-selectivity. Treatment of the cells with beta-GluCer increased the concentration of reactive oxygen species leading to cell cycle arrest and necrosis. Intraperitoneal administration of beta-GluCer significantly suppressed the growth of tumor implanted to the back of mice. beta-GluCer also induced antitumor immunity via the activation of NKT cells in vivo, and decreased the tumor metastasis of lymphoma cells. The present study thus demonstrated the antitumor activity of beta-GluCer in vivo, and discussed the mechanisms responsible for the growth inhibition.

Anti-adult T-cell leukemia effects of brown algae fucoxanthin and its deacetylated product, fucoxanthinol.

Adult T-cell leukemia (ATL) is a fatal malignancy of T lymphocytes caused by human T-cell leukemia virus type 1 (HTLV-1) infection and remains incurable. Carotenoids are a family of natural pigments and have several biological functions. Among carotenoids, fucoxanthin is known to have antitumorigenic activity, but the precise mechanism of action is not elucidated. We evaluated the anti-ATL effects of fucoxanthin and its metabolite, fucoxanthinol. Both carotenoids inhibited cell viability of HTLV-1-infected T-cell lines and ATL cells, and fucoxanthinol was approximately twice more potent than fucoxanthin. In contrast, other carotenoids, beta-carotene and astaxanthin, had mild inhibitory effects on HTLV-1-infected T-cell lines. Importantly, uninfected cell lines and normal peripheral blood mononuclear cells were resistant to fucoxanthin and fucoxanthinol. Both carotenoids induced cell cycle arrest during G(1) phase by reducing the expression of cyclin D1, cyclin D2, CDK4 and CDK6, and inducing the expression of GADD45alpha, and induced apoptosis by reducing the expression of Bcl-2, XIAP, cIAP2 and survivin. The induced apoptosis was associated with activation of caspase-3, -8 and -9. Fucoxanthin and fucoxanthinol also suppressed IkappaBalpha phosphorylation and JunD expression, resulting in inactivation of nuclear factor-kappaB and activator protein-1. Mice with severe combined immunodeficiency harboring tumors induced by inoculation of HTLV-1-infected T cells responded to treatment with fucoxanthinol with suppression of tumor growth, showed extensive tissue distribution of fucoxanthinol, and the presence of therapeutically effective serum concentrations of fucoxanthinol. Our preclinical data suggest that fucoxanthin and fucoxanthinol could be potentially useful therapeutic agents for patients with ATL.

Tumor specific cytotoxicity of glucosylceramide.

To develop a new taxon of anti-cancer agent with lower side effect, this study described a tumor selective cytotoxicity of glucosylceramide extracted from malt feed of beer brewing waste. Interpretation of (13)C- and (1)H-NMR spectra identified the chemical structure of major component of glucosylceramide as 1-O-beta-D: -glucopyranosyl-2(2'-hydroxyeicosanoylamino)-4,11-octadecadiene-1,3-diol. Selective cytotoxicity was studied with three pairs of normal and cancer cells: liver, skin and lung. The glucosylceramide selectively lowered the relative viability of cancer cells. Of the pairs, the selectivity was most pronounced with the liver cells, and, for this reason, further experiment was conducted with this pair of normal (CS-HC) and cancer cells (HepG2) to get more insight into the selective toxicity. The glucosylceramide significantly increased the cell population at G(2)/M phase in HepG2 cells, and also increased the numbers of apoptotic (sub-G(0)/G(1)) cells, but to much lesser extent compared with the increase in G(2)/M phase. Treatment of HepG2 cells with this agent selectively disrupted the mitochondrial membrane integrity without activation of caspase pathway to induce apoptosis. These findings suggested that the glucosylceramide specifically suppressed the growth of cancer cells by inhibiting cell renewal capacity rather than induction of apoptosis. The underlying mechanism for the selectivity remains to be answered in the forthcoming study.

Cerebral mature teratoma with malignant transformation in a 52-year-old man: a case report.

A case of adult teratoma with malignant transformation in a 52-year-old male is reported. We describe characteristic CT, histopathologic features and histogenesis of the tumor. Gradual onset of cerebral signs and symptoms was considered to be due to the tumor originating from the silent area of the frontal lobe of the brain and possessing a slowly progressive growth character.

Incorporation of branched-chain fatty acid into cellular lipids and caspase-independent apoptosis in human breast cancer cell line, SKBR-3.

BACKGROUND: 13-Methyltetradecanoic acid (13-MTD), an iso-C15 branched- chain saturated fatty acid, has been shown to induce apoptotic cell death of numerous human cancer cells. However, the mechanism for the induction of apoptosis has not been fully understood. This study described the incorporation of 13-MTD into cellular lipid of SKBR-3 breast cancer cells and apoptosis related event to gain more insight into the mechanism action of this fatty acid. RESULTS: Treatment of SKBR-3 cells with 13-MTD lowered the cell viability and induced apoptosis. Proportion of 13-MTD in the glycerolipids increased to saturation level within 6 hours. Triacylglycerol contained 13-MTD in higher concentration than phospholipid with positional preference to sn-2. 13-MTD caused no changes in the caspase activity and its gene expression. Furthermore, addition of caspase-inhibitor to culture medium did not prevent the cells from the cytotoxicity of 13-MTD. No-increase in the cellular calcium level was also noted with 13-MTD treatment. However, 13-MTD disrupted the mitochondrial integrity in 4 hours, and increased the nuclear translocation of apoptosis inducing factor. CONCLUSION: These results showed that 13-MTD disrupted the mitochondrial integrity, and induced apoptosis via caspase-independent death pathway.

Benign symmetric lipomatosis in a 73-year-old man: a case report with a brief review of the literature.

Benign symmetric lipomatosis (BSL) is a rare disease which is characterized by symmetric diffuse deposition of mature fat tissue and considered to originate in brown fat. A case of benign symmetric lipomatosis in a 73-year old man is presented. He has been treated for alcoholic abuse in mental hospitals several times and referred to our hospital for evaluation and treatment of the subcutaneous tumor in the neck, bilateral supraclavicular areas, bilateral upper arms, anterior chest wall, back, and the abdomen. Resected tumors had ill-defined margin and smooth surface. Histologically, the tumor was composed of diffuse proliferation of mature fat tissues, focal myxoid change and spindle cell proliferation. A histopathological diagnosis of lipomatosis with focal spindle cell proliferation was made. Although several hypotheses of this disease have been postulated, the present case suggested that the etiology of BSL is closely related with alcoholic abuse-induced metabolic disorder and deteriorated function of adipocytes due to specific location of tumor.

Effect of branched-chain fatty acids on fatty acid biosynthesis of human breast cancer cells.

This paper describes the antitumoral activity of branched-chain fatty acid (BCFA) in human breast cancer cells with an emphasis on its effect on fatty acid biosynthesis. First, the relationship between chain-length and antitumoral activity was studied. The highest activity was observed with iso-16:0, and the activity decreased with increase or decrease of the chain-lengths from C16:0. Anteiso-BCFA, as well as iso-series, was cytotoxic to the breast cancer cells. Cytotoxicity of BCFA was comparable to that of conjugated linoleic acid known as antitumoral fatty acid. Incubation of breast cancer cells with BCFA (13-methyltetradecanoic acid) significantly reduced the [14C] acetate incorporation into free fatty acid and fatty acid esters, showing the inhibition of fatty acid biosynthesis by BCFA. Examination of substrate level effect found that BCFA slightly inhibited fatty acid synthetase and acetyl-CoA carboxylase, and significantly the glucose-6-phosphate dehydrogenase which was the main NADPH generating system in breast cancer cells. The present study thus suggests that BCFA synthetically lowers the fatty acid biosynthesis by reducing the precursors, in addition to its direct inhibitory effect on fatty acid synthetase.

Analysis of microsatellite instability and loss of heterozygosity in human aortic atherosclerotic lesions.

The aim of the present study is to investigate whether microsatellite instability (MSI) and loss of heterozygosity (LOH) are involved in atherogenesis. Paraffin-embedded tissues of thoracic and abdominal aortas were collected from 29 non-neoplastic autopsied cases. We selected two types of atherosclerotic lesions including fibrocellular intimal thickening and uncomplicated atheromatous lesions, and analyzed two sites such as the aortic tunica intima and tunica media in each case. The frequencies of MSI and LOH were highest in the aortic tunica intima of atheromatous lesions and lowest in the aortic tunica media of fibrocellular intimal thickening lesions. Our results suggest that genetic instabilities such as MSI and LOH may be involved in the development of atherosclerotic lesions.

Conjugated linoleic acid (CLA) inhibits fatty acid synthetase activity in vitro.

This paper describes the in vitro effect of conjugated linoleic acid (CLA) on fatty acid biosynthesis. Among the rat liver enzymes involved in fatty acid biosynthesis, fatty acid synthetase (FAS) showed the largest activity fluctuation with the types of fatty acids. Of the fatty acids, CLA was the most potent inhibitor of FAS, and the 9c, 11t-rather than the 10t, 12c-isomer showed greater inhibition. CLA also significantly lowered the incorporation of [14C]-acetate into phospholipid in breast cancer cells, supporting the view that CLA inhibits fatty acid biosynthesis through the interaction with FAS.