Comprehensive Metabolomic Comparison of Five Cereal Vinegars Using Non-Targeted and Chemical Isotope Labeling LC-MS Analysis.

Vinegar is used as an acidic condiment and preservative worldwide. In Asia, various black vinegars are made from different combinations of grains, such as Sichuan bran vinegar (SBV), Shanxi aged vinegar (SAV), Zhenjiang aromatic vinegar (ZAV), and Fujian Monascus vinegar (FMV) in China and Ehime black vinegar in Japan (JBV). Understanding the chemical compositions of different vinegars can provide information about nutritional values and the quality of the taste. This study investigated the vinegar metabolome using a combination of GC-MS, conventional LC-MS, and chemical isotope labeling LC-MS. Different types of vinegar contained different metabolites and concentrations. Amino acids and organic acids were found to be the main components. Tetrahydroharman-3-carboxylic acid and harmalan were identified first in vinegar. Various diketopiperazines and linear dipeptides contributing to different taste effects were also detected first in vinegar. Dipeptides, 3-phenyllactic acid, and tyrosine were found to be potential metabolic markers for differentiating vinegars. The differently expressed pathway between Chinese and Japanese vinegar was tryptophan metabolism, while the main difference within Chinese vinegars was aminoacyl-tRNA biosynthesis metabolism. These results not only give insights into the metabolites in famous types of cereal vinegar but also provide valuable knowledge for making vinegar with desirable health characteristics.

[Physicians' Directions about Artificial Hydration and Nutrition at End-of-Life].

PURPOSE: To conduct a questionnaire survey to elicit physicians' directions about artificialhydration and nutrition at end-oflife. METHODS: A questionnaire was sent to 2,773 randomly selected Japanese physicians who were registrants of Rakuten Research Inc.(Tokyo, Japan). Of the respondents, 500 returned the questionnaire, and their reposes were analyzed. The Fisher's exact test was used to compare the respective responses. RESULTS: Of the 500 physicians, 22.4%(112)chose to discontinue artificialhydration and nutrition for those who are at terminalstage and could no longer communicate. On the other hand, 20.8%(104/500)of the physicians chose to continue artificialhydration and nutrition for those who were confined to bed and had the tendency to self-remove drip infusion or other tubes. Significantly more physicians who were working in hospitals than in private clinics chose to continue artificial hydration and nutrition(27% vs 3%; p<0.00001). CONCLUSION: Our findings revealed that 20.8% of physicians did not know about the latest evidence of end-of-life care to uphold patient dignity until the end-of-life. As our results might be different from the practice in other facilities located abroad, we must adopt prudent guidelines for palliative and end-of-life care and enlighten physicians and medical students regarding the current guidelines.

Intra-Abdominal Complications after Curative Gastrectomies Worsen Prognoses of Patients with Stage II-III Gastric Cancer.

BACKGROUND: Postoperative complications have been shown to worsen prognoses of various cancer types. METHODS: We retrospectively analyzed 265 patients with stage II-III gastric cancer who underwent curative gastrectomies between 1991 and 2010 at Tottori University Hospital to determine the effect of postoperative intra-abdominal complication (IAC) on prognosis. RESULTS: Of the 265 patients, 38 (14.3%) developed postoperative IACs of grade ≥ 2, of whom significantly more patients were male. Patients in the IAC group were significantly older than patients in the non-complication (NC) group. The NC group had significantly better survival than did the IAC group (P < 0.0001). Within the IAC group, 5-year survival rates did not significantly differ between patients with infectious complication subgroup (24.6%) and the non-infectious subgroup (46.2%). Grade of complication was not related to prognosis. Lengths of time before starting adjuvant chemotherapy (AC) after surgery were significantly longer for the IAC group (55.3 ± 34.7 days) than for the NC group: (26.6 ± 11.9 days; P = 0.0023). Prognosis of patients who took AC within 6 weeks after surgery tended to be better than that of patients who took AC > 6 weeks after surgery (P = 0.071). In multivariate analysis, IAC was an independent predictor of prognosis, as were age, invasion depth, and lymph node metastasis. CONCLUSION: Postoperative IACs were related to poorer survival for patients with stage II-III gastric cancer.

Anti-Proliferation Activity of Fucoidan in MKN45 Gastric Cancer Cells and Downregulation of Phosphorylated ASK1, a Cell Cycle-Regulated Kinase.

BACKGROUND: Fucoidan is a high-molecular polysaccharide whose main constituent is sulfated fucose. We specifically focused on the anti-proliferation activity of fucoidan and examined the underlying mechanism in MKN45 gastric cancer cells. METHODS: MKN45 cell proliferation was analyzed by BrdU assay and fucoidan cytotoxicity was examined by LDH and clonogenic assays. The Agilent Human microarray kit was used to identify upregulated and downregulated genes in response to fucoidan, and western blot analyses evaluated cell cycle proteins. RESULTS: Fucoidan impeded the MKN45 cell cycle by approximately 50%, and inhibited cell proliferation.LDH assays showed no immediate cytotoxic effects of fucoidan at 24 h exposure, however longer time courses revealed cell growth inhibition at 4 days in a dose-dependent manner. Microarray analysis identified MAP3K5, or ASK1 (apoptosis signal-regulating kinase),which was upregulated by 1.38-fold upon fucoidan treatment.Fucoidan increased ASK1 protein levels, while reducing phosphorylated ASK1 levels. Reduction of ASK1 by siRNA decreased proliferation of MKN45 cells. CONCLUSION: Our findings show that fucoidan may suppress cellular proliferation and DNA synthesis in MKN45 cells by suppressing the ASK1-p38 signaling pathway through reduction of phosphorylated ASK1 levels.

Effectiveness of the LigaSure Small Jaw Vessel-Sealing System in Hepatic Resection.

BACKGROUND: In hepatic resection for liver tumors, reducing operation time and blood loss are important for postsurgical complication prevention. This study aimed to compare the safety and efficacy of the LigaSure Small Jaw (Covidien, Boulder, CO) with those of the Cavitation Ultrasonic Surgical Aspirator (CUSA) system (Integra Life Sciences, Plainsboro, NJ) in hepatic surgery. METHODS: We enrolled 102 patients with liver tumors, of whom 51 underwent liver resection with the CUSA (CUSA group) between March 2004 and April 2011. Another 51 underwent resection with the LigaSure Small Jaw (LS group) between June 2011 and July 2012. We stratified patients by time period depending on the instrument used, and compared operative duration; intraoperative bleeding; and postoperative liver function and complication rate. RESULTS: Total operation time (mean ± SD) was significantly shorter in the LS group than in the CUSA group (358.8 ± 91.7 versus 460.6 ± 146.1 min, P < 0.001). Blood loss was not significantly different between the 2 groups. Frequency of postoperative complications was lower, but not significantly, in the LS group. CONCLUSION: The LigaSure Small Jaw may allow a shorter total operative duration than the CUSA device.

Usefulness of palliative prognostic score in the treatment of patients with non-resectable gastric cancer.

The aim of this study was to evaluate the clinical usefulness of the palliative prognostic (PaP) score in patients with non-resectable advanced gastric cancer. The PaP score was calculated prior to each course of chemotherapy in 44 consecutive patients with non-resectable advanced gastric cancer between 2003 and 2010 at the Tottori University Hospital, Yonago, Japan. The prognosis was evaluated according to the PaP score and the different chemotherapeutic agents. The median survival time (MST) was 10 months. The PaP score classified the heterogeneous patient sample into three isoprognostic groups with regard to the possibility of a 1-month survival period, with 28 patients in group A (>70% chance), 12 in group B (30-70% chance) and 4 in group C (<30% chance). The MST of the three groups was 11, 3 and 1 months for group A, B and C, respectively. In group A, chemotherapeutic regimens did not affect patient survival, although the docetaxel regimen prolonged survival of patients in group B. In conclusion, the PaP score may be useful in selecting the best chemotherapeutic regimen in patients with non-resectable gastric cancer.

Phylogenomic and domain analysis of iterative polyketide synthases in Aspergillus species.

Aspergillus species are industrially and agriculturally important as fermentors and as producers of various secondary metabolites. Among them, fungal polyketides such as lovastatin and melanin are considered a gold mine for bioactive compounds. We used a phylogenomic approach to investigate the distribution of iterative polyketide synthases (PKS) in eight sequenced Aspergilli and classified over 250 fungal genes. Their genealogy by the conserved ketosynthase (KS) domain revealed three large groups of nonreducing PKS, one group inside bacterial PKS, and more than 9 small groups of reducing PKS. Polyphyly of nonribosomal peptide synthase (NRPS)-PKS genes raised questions regarding the recruitment of the elegant conjugation machinery. High rates of gene duplication and divergence were frequent. All data are accessible through our web database at http://metabolomics.jp/wiki/Category:PK.

Database for crude drugs and Kampo medicine.

A wiki-based repository for crude drugs and Kampo medicine is introduced. It provides taxonomic and chemical information for 158 crude drugs and 348 prescriptions of the traditional Kampo medicine in Japan, which is a variation of ancient Chinese medicine. The system is built on MediaWiki with extensions for inline page search and for sending user-input elements to the server. These functions together realize implementation of word checks and data integration at the user-level. In this scheme, any user can participate in creating an integrated database with controlled vocabularies on the wiki system. Our implementation and data are accessible at http://metabolomics.jp/wiki/.

Exposure to extremely low frequency magnetic fields affects insulin-secreting cells.

To evaluate the effects of extremely low frequency magnetic field (ELFMF) on beta-cell survival and function, we cultured a hamster-derived insulin-secreting cell line (HIT-T15), which exhibits responsiveness to glucose in a semi-physiological range, under exposure to sham and ELFMF conditions, and assessed cell survival and function. We used our previously developed ELFMF exposure unit (a sinusoidal magnetic field at a frequency of 60 Hz, 5 mT) to culture cells under exposure to ELFMF conditions. We found that exposure to ELFMF for 5 days in the absence of glucose increased cell number, exposure for 2 days in the absence of glucose and for 5 days with 100 mg/dl glucose increased the insulin secretion to the culture medium, and exposure for 2 and 5 days with 40 and 100 mg/dl glucose increased intracellular insulin concentration in HIT-T15 cells. The increase in cell number under apoptotic culture conditions by exposure to ELFMF could lead to new therapeutic concepts in the treatment of diabetes. The ELFMF-induced increase in intracellular insulin concentration could be utilized to develop culture conditions to enhance intracellular insulin concentration in insulin-secreting cells that would be useful for cell transplantation to cure diabetes mellitus.

Radiation-induced reduction of osteoblast differentiation in C2C12 cells.

Therapeutic radiation causes bone damage and may increase fracture risks in treatment for head-and-neck cancer and in pelvic irradiation. These properties can also be used for prevention of heterotopic ossification in hip arthroplasty. To evaluate the effects of ionizing radiation on osteoblast differentiation, C2C12 cells were directed into an osteogenic lineage by treatment with a combination of bone morphogenic protein 2 (BMP-2) (100 ng/ml) and heparin (30 mug/ml) 6 h after irradiation (2 and 4 Gy). Osteoblast differentiation was evaluated based on alkali phosphatase (ALP) activity and expression of mRNA encoding ALP and collagen type I. Ionizing radiation suppressed the growth of C2C12 cells and decreased expression of ALP and collagen type I mRNAs with concomitant reduction of the ALP activity. Although further studies are needed to elucidate the molecular mechanism, our findings suggest that ionizing radiation at therapeutic doses interferes with bone formation by reducing ALP activity and expression of mRNA encoding ALP and collagen type I.

Correlation between the severity of clinicopathological parameters and whole blood interferon-alpha production capacity in active phase IgA nephropathy patients.

BACKGROUND/AIMS: Patients with IgA nephropathy (IgA-N) are thought to have immune system disorders that frequently result in high serum IgA levels and a relatively high susceptibility to upper respiratory infections. AIMS: To clarify the influence of the specific immune response of IgA-N patients on the clinicopathological features of the disease, we measured the whole-blood-producing capacity of interferon-alpha (IFNalpha-PC). We then compared these findings with clinical and histopathological parameters, including tissue macrophage infiltration, during both histologically active and latent phases. PATIENTS AND METHODS: Fifty-one inpatients with IgA-N and 70 healthy controls were examined. According to the histological findings, 32 patients had disease in the active phase (AP), and 19 were in the latent phase (LP). RESULTS: In AP patients, IFNalpha-PC showed positive correlations to serum creatinine, blood urea nitrogen, serum beta2-microglobulin (s-beta2MG), urinary total protein (U-TP), and urinary beta2MG, in addition to the number of infiltrated macrophages per area of interstitium. In LP patients, negative correlations were shown between IFNalpha-PC and s-beta2MG, U-TP, and U-N-acetyl-beta-D-glucosaminidase. CONCLUSION: A significant positive relationship exists between IFNalpha-PC and the clinicopathological parameters of deteriorated renal lesions in the AP but not in the LP. Thus, the immune status influencing the functional damage may differ between these two phase.