Macrophi Inc.

Kagawa ken, Japan

Macrophi Inc.

Kagawa ken, Japan
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Kobayashi Y.,Kagawa University | Inagawa H.,Kagawa University | Inagawa H.,Technology Research Association | Inagawa H.,Niigata University of Pharmacy and Applied Life Sciences | And 9 more authors.
Anticancer Research | Year: 2017

Background/Aim: Recent studies reported that lipopolysaccharide (LPS) exhibits beneficial effects on prevention of immune-related diseases by activating macrophages. We previously demonstrated that pre-treatment with LPS derived from Pantoea agglomerans (LPSp) activated amyloid (A) phagocytosis in mouse primary microglia. In the present study, we further examined the promotory effect on phagocytosis of phagocytic particles in the C8-B4 microglia cell line. Materials and Methods: Phagocytic analysis of C8-B4 cells was evaluated using phagocytic particles (latex beads or HiLyte™ Fluor 488-conjugated A1-42). Results: The phagocytic activity of latex beads was dependent on the concentration of beads and incubation time. LPSp, at as low as 100 pg/ml, significantly increased phagocytosis against the beads. In the experiment of A1-42 phagocytosis, LPSp significantly increased A phagocytic activity. Conclusion: LPSp treatment was confirmed to enhance A1-42 phagocytosis by mouse microglia. It is suggested that the use of LPSp may be a potential promising candidate for the prevention of Alzheimer's disease.


Kadowaki T.,Kagawa University | Inagawa H.,National Fisheries University | Inagawa H.,Macrophi Inc. | Inagawa H.,Tokyo University of Science | And 9 more authors.
Anticancer Research | Year: 2011

Background and Aim: Lipopolysaccharide derived from a symbiotic bacterium in wheat (Pantoea agglomerans, LPSp) has shown multiple positive effects, such as prophylactic, antiallergic and antitumour effects, without serious side-effects. LPSp has differential biological activities in comparison to other LPS, such as those from Escherichia coli (LPSe). The only difference between LPSp and LPSe is in the O-antigen polysaccharide structure (O-PS). This led us to the hypothesis that the O-PS structure would seem to participate in biological activities. Thus, the characterization of properties of O-PS in LPS is of the utmost importance for understanding cell activation in the maintenance of homeostasis. However, little is known about the correlation between the O-PS structure of LPS and its biological activities. In this study, we extracted LPS derived from a symbiotic bacterium in rice (strain A46, related species of Pantoea), which has a long history of use in foods, and investigated its putative structures and functions. Materials and Methods: LPS derived from strain A46 was prepared using a hot phenol extraction method. The properties of LPS-A46 were analysed by thin-layer chromatography, Tricine SDS-PAGE and Western blotting. The function of LPS-A46 was analyzed by quantative real-time PCR and flow cytometry using THP-1 cells and Peripheral blood mononuclear cell (PBMC) derived macrophages. Results: In Tricine SDS-PAGE, high molecular mass LPS-A46 had a molecular mass lower than that of LPSp. In Western blotting, LPS-A46 reacted with lipid A antibody but did not react with an O-PS antibody of LPSp. In comparison to other LPS, LPS-A46 induced a differential cytokine gene expression profile in THP-1 cells and PBMC-derived macrophages. Conclusion: The present study suggests that LPS derived from symbiotic bacterium in rice is a bioactive functional LPS which may have different functional activities compared to other types of LPS.


Wakame K.,Hokkaido University | Komatsu K.-I.,Hokkaido University | Inagawa H.,Kagawa University | Inagawa H.,Technology Research Inc | And 2 more authors.
Anticancer Research | Year: 2015

Background/Aim: Pantoea agglomerans LPS (immunopotentiator from Pantoea agglomerans 1: IP-PA1) has been reported to have anti-inflammatory effects in in vitro and in vivo models. The aim of the present study was to investigate the effects of orally-administered IP-PA1 on atopic dermatitis (AD) symptoms induced by Dermatophagoides farinae body extract (DFE) in NC/Nga mice. Materials and Methods: Using the NC/Nga AD murine model, mice were orally administered 0.1% (High) or 0.01% (Low) water-containing IP-PA1. Skin lesion assessment and blood collection from the caudal vein was performed on days 0, 7, 21 and 31. On day 31, all mice were sacrificed and blood, skin, spleen, as well as intestine samples, were obtained. Results: Assessment score of the skin lesion and serum immunoglobulin E (IgE) level of both IP-PA1 groups were significantly lower than that of the DFE group on days 14 and 21. The serum periostin and thymus and activation-regulated chemokine (TARC) level of IP-PA1-Low group was significantly lower than that of the DFE group on day 31. On histological examination of the skin, hyperplasia of epidermal and dermal layers and infiltration of inflammatory cells were suppressed by IP-PA1 administration. Deposition of periostin was observed in the DFE group skin tissue. Moreover, the CD4+/CD8+ ratio of splenic T-cells increased by IP-PA1 administration. Conclusion: IP-PA1 administration may have an inhibitory effect on AD skin lesions.


Kadowaki T.,Kagawa University | Ohno S.,Kagawa University | Taniguchi Y.,Kagawa University | Taniguchi Y.,Tokushima Bunri University | And 7 more authors.
Anticancer Research | Year: 2013

Background: Lipopolysaccharide derived from Pantoea agglomerans (LPSp) mainly consists of two aggregates, the high-molecular aggregate (HMM-LPSp) and the low-molecular aggregate (LMM-LPSp). The structural differences between HMM-LPSp and LMM-LPSp seem to depend on the length of the O-antigen polysaccharide because the lipid A regions of the two fractions are quite similar. In this study, we examined the biological activity of LPSp focusing on the O-antigen polysaccharide using HMM-LPSp and LMMLPSp under serum-free conditions. Materials and Methods: The binding of LPSp to RAW264.7 cells under serum-free conditions was analyzed by flow cytometry using LPSpconjugated fluorescein isothiocyanate (FITC-LPSp). The biological activities of HMM-LPSp and LMM-LPSp under serum-free conditions were evaluated by the nitric oxide production. Results: FITC-LPSp showed higher fluorescence intensity under serum-free than serum-containing conditions. HMM-LPSp induced higher nitric oxide production than LMMLPSp under serum-free conditions. Conclusion: The present study indicates that the reactivity of LPSp is affected by the Oantigen polysaccharide under serum-free conditions.


Kadowaki T.,Kagawa University | Inagawa H.,Kagawa University | Inagawa H.,Macrophi Inc. | Inagawa H.,Tokyo University of Science | And 8 more authors.
In Vivo | Year: 2011

Aims: Anti-lipopolysaccharide factor (ALF) is an antimicrobial peptide (AMP) and a key effector molecule of the innate immune system in crustaceans. However, little is known about the role of its indirect killing against bacteria. The possible regulatory role of this peptide (M-ALF) in kuruma prawns, Marsupenaeus japonicus, was investigated. Materials and Methods: The activities of M-ALF were investigated by antimicrobial activity in vitro and by experimental infection Vibrio penaeicida in vivo with ALF-knock down in kuruma prawn by systemically silencing M-ALF gene through the injection of gene-specific long double-stranded RNA with RNA interference. Results: Synthetic M-ALF had no direct antimicrobial activity against V. penaeicida, whereas ALF-silenced kuruma prawns had significantly higher mortality than untreated prawn after V. penaeicida infection. The data provide compelling evidence that M-ALF plays an indirect protective role against V. penaeicida infection, suggesting the idea that ALF acts as a cytokine-like regulatory molecule, as well as an effector molecule. Conclusion: M-ALF has no direct activity against V. penaeicida, but might be a key molecule in cytokine-like gene regulation in crustaceans.


Kato M.,Hanakoganei Clinic | Goto S.,Chiyoda Corporation | Soma G.-I.,Tokushima Bunri University | Soma G.-I.,Macrophi Inc. | And 2 more authors.
Anticancer Research | Year: 2010

We describe the case of a patient with terminal adenocarcinoma of the lung with no response to lymphokine-activated killer (LAK) cell therapy alone who distinctly responded clinically to a combination of high-dose glucocorticoids (GC) and LAK cell therapy, administered by chance. However, after decreasing the dose of GC because of gastrointestinal bleeding caused by the high-dose treatment, there was no response on restarting GC plus LAK cell therapy. The clinical course of this case strongly suggests that local inflammation in the vicinity of tumors should be adequately suppressed in patients with advanced cancer who receive immunotherapy.


PubMed | Toyo Rice Corporation, Macrophi Inc., Nagoya Keizai University, Niigata University of Pharmacy and Applied Life Sciences and Kagawa University
Type: Journal Article | Journal: Anticancer research | Year: 2016

Oral ingestion of lipopolysaccharide (LPS) has been shown to be effective in diseases prevention. Brown rice contains large amounts of LPS not actively consumed because of bad taste. Recently, a new type of brown rice with its wax layer removed has been produced. In this report, we measured the LPS content of this dewaxed rice and evaluated the function of innate immune activation on macrophages.Dewaxed brown rice and polished rice were prepared using the Saika-style rice polishing process. LPS content extracted using hot water from this sample was evaluated by the Limulus reaction and the activation of macrophage RAW246.7 cells was evaluated by nitric oxide (NO) production. In addition, toll-like receptors (TLRs) 2-, 4- and 9-induced human embryonic kidney (HEK) 293 cells were used for the confirmation of the activated pathway.Mean LPS content in the 15 types of dewaxed brown rice was found to be 6.42.6 g/g, while that of brown rice was 10.94.3 g/g. The extract of dewaxed brown rice induced significant amounts of NO by RAW246.7 cells, while production was reduced to 1/6 by adding polymyxin B. The macrophage activating effect of dewaxed brown rice was 79- and 51-times higher than that of polished rice in TLR4- and 2-induced HEK 293 cells.LPS content in dewaxed brown rice was found to be able to activate macrophages. This rice activated macrophages mainly via the TLR4 and, to a lesser extent, TLR2 pathways. It is suggested that dewaxed brown rice can be considered effective in allergy and cancer prevention.


PubMed | Hokkaido Pharmaceutical University School of Pharmacy, Kagawa University, Macrophi Inc., Niigata University of Pharmacy and Applied Life Sciences and Scare Crow Inc.
Type: Journal Article | Journal: Anticancer research | Year: 2016

The lipopolysaccharide (LPS)-like compound derived from Pantoea agglomerans (immunopotentiator from Pantoea agglomerans 1 (IP-PA1)) has been used not only as dietary supplement or cosmetic for humans, but also by Japanese veterinarians as an anti-tumor, anti-allergy, keep a fine coat of fur and hair growth-promoting functional food for dogs and cats. In the present study, we focused on the hair growth-promoting effects of IP-PA1 on a hair-shaved animal model and its mechanism of action. We also investigated its potential on gene expression after stimulating human dermal papilla cells with IP-PA1.The hair on the back of a C3H/HeN mouse was shaved and IP-PA1 was orally administered or applied to the skin. The status of hair growth was observed and recorded for 14 days. Skin was collected and histological tissue examination was performed with respect to hair growth status using hematoxylin and eosin staining. After IP-PA1 administration (2 and 10 g/ml) to human dermal papilla cell culture system for 24 h, fibroblast growth factor-7 (FGF-7) and vascular endothelial growth factor (VEGF) mRNA expression were measured using real-time polymerase chain reaction (PCR) analysis.IP-PA1, when given orally, showed a tendency to promote hair growth in mice. In addition, skin application also significantly promoted hair growth, while histopathological examinations further demonstrated hair elongation from dermal papilla cells. In the human dermal papilla cell culture system, significant FGF-7 and VEGF mRNA expressions were observed (p<0.05).An underlying mechanism of gene expression by which IP-PA1 promotes hair growth was suggested to be different from that of medicine and traditional hair tonics, such as minoxidil and adenosine.


Patent
Soma, Res Group, Inc. and Macrophi Inc. | Date: 2012-08-22

A disease resistance capacity of vegetables, grasses and flowers, etc. , is enhanced and their survival rates are improved by growing plants in an appropriate coexistence of a glycolipid derived from Pantoea agglomerans or a composition containing the glycolipid. Thereby, a method of augmenting disease resistance of vegetables, grasses and flowers, etc. , and growing agricultural crops with high quality is provided, and further a novel plant growing agent and plant disease resistance inducer that enables this are provided. As a result, it becomes possible to cultivate agricultural crops or increase productivity with as little use of agricultural chemicals as possible. Even if an amount of the agricultural chemical is reduced, a cultivation success rate of agricultural crops can be enhanced and immunity of the plant body can be enhanced even where useful soil bacteria are not frequently present. Thus, new types of agricultural crop businesses such as contract cultivations by means of organic cultivation, agricultural chemical-free cultivation and hydroponic cultivation are promoted.


It has been found that a limulus-positive glycolipid is present in xanthan gum derived from Xanthomonas, which has been commercially available and eaten for many years, and this was purified, and it has been found that this limulus-positive glycolipid has an immunopotentiation effect. A method for safely and inexpensively producing the limulus-positive glycolipid containing an immunopotentiator at high concentrations is provided. The method for producing the limulus-positive glycolipid of the present invention comprises extracting the limulus-positive glycolipid from xanthan gum. A limulus-positive glycolipid composition containing the limulus-positive glycolipid can be used for various applications such as pharmaceuticals, pharmaceuticals for animals, quasi drugs, cosmetics, foods, functional foods, feedstuff and bath agents.

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