Gravi-Agari™

Gravi-Agari™ — (tonic) — Rich in Beta glucans and selective cytotoxic Annonacea acetogenins, this formula is naturally derived from full spectrum Agaricus Sun Supreme™ mushroom and from the whole leaves of Amazon Annona muricata (graviola). The fruiting body of Agaricus blazei has become Japan’s primary immune supporting herb, most frequently recommended for adjunct treatment and support, and Graviola is popular for its effectiveness against all types of cancer. Studies show these botanicals are cytotoxic to numerous t-cell lines.1,4,6,7,9,10,11,13,16,17,18,19, 20,21,22,24,25,26 Graviola is an effective free radical scavenger, having favorable effects on patients with cancer, augmenting its therapeutic benefits.2,12 It is also antiviral, antibacterial, antifungal, and antiprotozoal (parasites).3,4,5,8, Agaricus is an effective immunomodulator and antimutagenic (cellular protector), having a protective effect against chemically induced DNA damage, and shows natural killer cell activity and improved quality of life with cancer patients undergoing chemotherapy.12,14,15,16,18,19,21,23,25,27

19. Natural killer cell activity and quality of life were improved by consumption of a mushroom extract, Agaricus blazei Murill Kyowa, in gynecological cancer patients undergoing chemotherapy
W.-S. Ahn*1, D.-J. Kim†, G.-T. Chae‡, J.-M. Lee*, S.-M. Bae§, J.-I. Sin§, Y.-W. Kim§, S.-E. Namkoong*& I. P. Lee**Department of Obstetrics and Gynecology; †Department of Medical Statistics; ‡Institute of Chronic Disease; and §Catholic Research Institutes of Medical Science, College of Medicine, The Catholic University of Korea, Seoul, South Korea.
International Journal of Gynecological Cancer 14 (4), 589–594.
Volume 14 Issue 4 Page 589 - July 2004
www.blackwell-synergy.com

1. Antiprotozoal and cytotoxic activities in vitro of Colombian Annonaceae.
Osorio E, Arango GJ, Jiménez N, Alzate F, Ruiz G, Gutiérrez D, Paco MA, Giménez A, Robledo S.
Grupo de Investigación en Sustancias Bioactivas (GISB), Facultad de Química Farmacéutica, Corporación de Patologías Tropicales, Universidad de Antioquia, A.A. 1226, Medellín, Colombia. josorio@farmacia.udeaeduco.
J Ethnopharmacol. 2007 May 22;111(3):630-5. Epub 2007 Jan 18.
Ethnobotanical and chemotaxonomical studies for antiparasitic activity of Colombian Annonaceae were carried out. In vitro antiprotozoal activity of 36 extracts obtained from six different species was determined against promastigotes of three Leishmania species, epimastigotes of Trypanosoma cruzi and both chloroquine sensitive (F32) and resistant (W2) Plasmodium falciparum. Cytotoxic activity was evaluated in U-937 cells. Active extracts were selected according their selectivity index (SI). Extracts from Annona muricata, Rollinia exsucca, Rollinia pittieri and Xylopia aromatica were active against Leishmania spp. and Trypanosoma cruzi showing IC50 values lower than 25 microg/ml. Hexane extract from Rollinia pittieri leaves was the most selective against Trypanosoma cruzi and Leishmania spp. (IS=10 and 16, respectively). The extracts from Desmopsis panamensis, Pseudomalmea boyacana, Rollinia exsucca and Rollinia pittieri showed good antiplasmodial activity (IC50 < 10 microg/ml). No correlation between antiplasmodial activity and inhibition of beta-hematin production was found. The present study gives specific and useful information about antiprotozoal and cytotoxic activities of some Annonaceae extracts. Results presented here also demonstrate which plants and/or plant parts could be useful in the treatment of leishmaniasis, Chagas’ disease and malaria.
PMID: 17296281 [PubMed - indexed for MEDLINE]

2. In vitro antioxidant studies in leaves of Annona species.
Baskar R, Rajeswari V, Kumar TS.
Department of Biotechnology, Kumaraguru College of Technology, Coimbatore 641 006, India. bhubaski@rediffmail.com
Indian J Exp Biol. 2007 May;45(5):480-5.
Antioxidant potential of leaves of three different species of Annona was studied by using different in vitro models eg., 1,1-diphenyl-2-picryl hydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS), nitric oxide, superoxide, hydroxy radical and lipid peroxidation. The ethanolic extract of A. muricata at 500 microg/ml showed maximum scavenging activity (90.05%) of ABTS radical cation followed by the scavenging of hydroxyl radical (85.88%) and nitric oxide (72.60%) at the same concentration. However, the extract showed only moderate lipid peroxidation inhibition activity. In contrast, the extract of A. reticulata showed better activity in quenching DPPH (89.37%) and superoxide radical (80.88%) respectively. A.squamosa extract exhibited least inhibition in all in vitro antioxidant models excepting hydroxyl radical (79.79%). These findings suggest that the extracts of A. muricata possess potent in vitro antioxidant activity as compared to leaves of A. squamosa and A. reticulata suggesting its role as an effective free radical scavenger, augmenting its therapeutic benefits.
PMID: 17569293 [PubMed - indexed for MEDLINE]

3. Antibacterial activity of eight Brazilian annonaceae plants.
Takahashi JA, Pereira CR, Pimenta LP, Boaventura MA, Silva LG.
Departamento de Química -- ICEx -- Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, CEP 31270-901, Belo Horizonte, MG, Brazil. jacfab@dedalus.lcc.ufmg.br
Nat Prod Res. 2006 Jan;20(1):21-6.
Sixteen extracts, obtained from eight Brazilian plants of Annonaceae family, were screened for their antibacterial activity: Xylopia frutescens, X. aromatica, X. amazonica, X. benthamii, Annona ambotay, A. crassiflora, A. muricata and A. cherimolia. Amongst the investigated extracts, six showed antibacterial activity against at least one of the tested organisms at the concentration of 100 microg/mL. The most active extracts were those prepared from X. frutescens, X. amazonica, and A. ambotay. A phytochemical screening showed the presence of anonaceus acetogenins in some active extracts. Eleven diterpenoids were also tested for comparison purposes. Six were natural products, previously isolated from Xylopia sp. (kaurenoic, frutoic, xylopic, 15beta-hydroxy-kaurenoic and trachylobanic acids plus kaurenol) and five were derivatives of such compounds, obtained by esterification or reduction reactions. Trachylobanic acid showed antibacterial activity against B. subtilis and S. aureus.
PMID: 16286303 [PubMed - indexed for MEDLINE]

4. Antitumor and Antiviral Activity of Colombian Medicinal Plant Extracts
LA Betancur-Galvis/+, J Saez*, H Granados*, A Salazar**, JE Ossa
Laboratorio de Virología, Departamento de Microbiología y Parasitología, Facultad de Medicina *Departamento de Química **Departamento de Biología, Facultad de Ciencias Exactas y Naturales, Universidad de Antioquia, Apartado 1226, Medellín, Colombia 531 Mem Inst Oswaldo Cruz, Rio de Janeiro, Vol. 94(4): 531-535, Jul./Aug. 1999
Extracts of nine species of plants traditionally used in Colombia for the treatment of a variety of diseases were tested in vitro for their potential antitumor (cytotoxicity) and antiherpetic activity. MTT (Tetrazolium blue) and Neutral Red colorimetric assays were used to evaluate the reduction of viability of cell cultures in presence and absence of the extracts. MTT was also used to evaluate the effects of the extracts on the lytic activity of herpes simplex virus type 2 (HSV-2). The 50% cytotoxic concentration (CC50) and the 50% inhibitory concentration of the viral effect (EC50) for each extract were calculated by linear regression analysis. Extracts from Annona muricata, A. cherimolia and Rollinia membranaceal, known for their cytotoxicity were used as positive controls. Likewise, acyclovir and heparin were used as positive controls of antiherpetic activity. Methanolic extract from Annona sp. on HEp-2 cells presented a CC50 value at 72 hr of 49.6x103 g/ml. Neither of the other extracts examined showed a significant cytotoxicity. The aqueous extract from Beta vulgaris, the ethanol extract from Callisia grasilis and the methanol extract Annona sp. showed some antiherpetic activity with acceptable therapeutic indexes (the ratio of CC50 to EC50). These species are good candidates for further activity-monitored fractionation to identify active principles.
PMID: 10446015 [PubMed - indexed for MEDLINE]

5. Isolation and characterization of a lectin from Annona muricata seeds.
Damico DC, Freire MG, Gomes VM, Toyama MH, Marangoni S, Novello JC, Macedo ML.Departamento de Bioquimica, Instituto de Biologia, Universidade Estadual de Campinas, Campinas (SP), Brazil.
J Protein Chem. 2003 Nov;22(7-8):655-61.
A lectin with a high affinity for glucose/mannose was isolated from Annona muricata seeds (Annonaceae) by gel filtration chromatography on Sephacryl S-200, ion exchange chromatography on a DEAE SP-5 PW column, and molecular exclusion on a Protein Pak Glass 300 SW column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (PAGE) yielded two protein bands of approximately 14 kDa and 22 kDa. However, only one band was seen in native PAGE. The Mr of the lectin estimated by fast-performance liquid chromatography-gel filtration on Superdex 75 was 22 kDa. The lectin was a glycoprotein with 8% carbohydrate (neutral sugar) and required divalent metal cations (Ca2+, Mg2+, and Mn2+) for full activity. Amino acid analysis revealed a large content of Glx, Gly, Phe, and Lys. The lectin agglutinated dog, chicken, horse, goose, and human erythrocytes and inhibited the growth of the fungi Fusarium oxysporum, Fusarium solani, and Colletotrichum musae.
PMID: 14714732 [PubMed - in process]

6. New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata.
Liaw CC, Chang FR, Lin CY, Chou CJ, Chiu HF, Wu MJ, Wu YC.
Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan, Republic of China.
J Nat Prod. 2002 Apr;65(4):470-5.
Three new monotetrahydrofuran annonaceous acetogenins, muricin H (1), muricin I (2), and cis-annomontacin (3), along with five known acetogenins, annonacin, annonacinone, annomontacin, murisolin, and xylomaticin, were isolated from the seeds of Annona muricata. Additionally, two new monotetrahydrofuran annonaceous acetogenins, cis-corossolone (4) and annocatalin (5), together with four known ones, annonacin, annonacinone, solamin, and corossolone, were isolated from the leaves of this species. The structures of all new isolates were elucidated and characterized by spectral and chemical methods. These new acetogenins exhibited significant activity in in vitro cytotoxic assays against two human hepatoma cell lines, Hep G(2) and 2,2,15. Compound 5 showed a high selectivity toward the Hep 2,2,15 cell line.
PMID: 11975482 [PubMed - indexed for MEDLINE]

7. Novel cytotoxic annonaceous acetogenins from Annona muricata.
Chang FR, Wu YC.Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan, Republic of China.
J Nat Prod. 2001 Jul;64(7):925-31.
Seven new annonaceous acetogenins, muricins A-G (1-7), as well as five known compounds, a mixture of muricatetrocin A (8) and muricatetrocin B (9), longifolicin (10), corossolin (11), and corossolone (12), were isolated from the seeds of Annona muricata. The structures of all isolates were elucidated and characterized by spectral and chemical methods. These acetogenins showed significantly selective in vitro cytotoxicities toward the human hepatoma cell lines Hep G(2) and 2,2,15.
PMID: 11473425 [PubMed - indexed for MEDLINE]

8. Effect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes simplex virus.
Padma P, Pramod NP, Thyagarajan SP, Khosa RL.
Department of Pharmaceutics, IT, Banaras Hindu University, Varanasi, India.
Annona muricata (Annonaceae) and Petunia nyctaginiflora (Solanaceae) were screened for their activity against Herpes simplex virus-1 (HSV-1) and clinical isolate (obtained from the human keratitis lesion). We have looked at the ability of extract(s) to inhibit the cytopathic effect of HSV-1 on vero cells as indicative of anti-HSV-1 potential. The minimum inhibitory concentration of ethanolic extract of A. muricata and aqueous extract of P. nyctaginiflora was found to be 1 mg/ml.
PMID: 9687085 [PubMed - indexed for MEDLINE]

9. Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10R-annonacin-A-ones, from the leaves of Annona muricata.
Wu FE, Zhao GX, Zeng L, Zhang Y, Schwedler JT, McLaughlin JL, Sastrodihardjo S.
Department of Medicinal Chemistry and Pharmacognosy, School of Pharmacy and Pharmacal Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
J Nat Prod. 1995 Sep;58(9):1430-7.
In a continuation of our research on bioactive components from the leaves of Annona muricata, three novel monotetrahydrofuran Annonaceous acetogenins, namely, annomutacin [1], (2,4-trans)-10R-annonacin-A-one [2], and (2,4-cis)-10R- annonacin-A-one [3], have been identified. Their structures were deduced by ms, nmr, ir, and uv spectral and chemical methods, and the absolute configurations were determined by Mosher ester methodology. A known bioactive amide, N-p-coumaroyl tyramine, was also found. Compound 1 and the mixture of compounds 2 and 3 showed selective cytotoxicities against the human A-549 lung tumor cell line.

10. Studies on the chemical constituents of Annona muricata [Article in Chinese]
Yu JG, Gui HQ, Luo XZ, Sun L, Zhu P, Yu ZL.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100094.
Yao Xue Xue Bao. 1997 Jun;32(6):431-7.
Annonaceous acetogenin (or polyketide) is a kind of potential antineoplastic agents from Annonaceae plants. Two new acetogenins, Muricatalicin (I) and muricatalin (VI), a mesitoate of a new acetogenin, annonacin-B mesitoate (Vb), and three known acetogenins, annonacin (II), annonacin-A (III) and annonacin-10-one (IV) have been isolated from Annona muricata L. The structures and relative stereochemistry of I, VI and Vb were elucidated on the basis of spectral analysis and examination of their acetates and/or mesitoate.
PMID: 11596323 [PubMed - indexed for MEDLINE]

11. Primary mechanism of apoptosis induction in a leukemia cell line by fraction FA-2-b-ss prepared from the mushroom Agaricus blazei Murill.
Gao L, Sun Y, Chen C, Xi Y, Wang J, Wang Z.
School of Life Sciences, Lanzhou University.
Braz J Med Biol Res. 2007 Nov;40(11):1545-55.
Agaricus blazei Murill is a native Brazilian mushroom which functions primarily as an anticancer substance in transplanted mouse tumors. However, the mechanism underlying this function of A. blazei Murill remains obscure. The present study was carried out to investigate the effect of fraction FA-2-b-ss, an RNA-protein complex isolated from A. blazei Murill, on human leukemia HL-60 cells in vitro. Typical apoptotic characteristics were determined by morphological methods using DNA agarose gel electrophoresis and flow cytometry. The growth suppressive effect of fraction FA-2-b-ss on HL-60 cells in vitro occurred in a dose- (5-80 microg/mL) and time-dependent (24-96 h) manner. The proliferation of HL-60 cells (1 x 10(5) cells/mL) treated with 40 microg/mL of fraction FA-2-b-ss for 24-96 h and with 5-80 microg/mL for 96 h resulted in inhibitory rates ranging from 8 to 54.5%, and from 4.9 to 86.3%, respectively. Both telomerase activity determined by TRAP-ELISA and mRNA expression of the caspase-3 gene detected by RT-PCR were increased in HL-60 cells during fraction FA-2-b-ss treatment. The rate of apoptosis correlated negatively with the decrease of telomerase activity (r = 0.926, P < 0.05), but correlated positively with caspase-3 mRNA expression (r = 0.926, P < 0.05). These data show that fraction FA-2-b-ss can induce HL-60 cell apoptosis and that the combined effect of down-regulation of telomerase activity and up-regulation of mRNA expression of the caspase-3 gene could be the primary mechanism of induction of apoptosis. These findings provide strong evidence that fraction FA-2-b-ss could be of interest for the clinical treatment of acute leukemia.
PMID: 17934651 [PubMed - in process]

12. Measuring Perceived Effects of Drinking an Extract of Basidiomycetes Agaricus blazei Murill: A Survey of Japanese Consumers with Cancer.
Talcott JA, Clark JA, Lee IP.
BMC Complement Altern Med. 2007 Oct 29;7(1):32 [Epub ahead of print]
ABSTRACT: BACKGROUND: To survey cancer patients who consume an extract of the Basidiomycetes Agaricus blazei Murill mushroom (Sen-Sei-Ro) to measure their self-assessment of its effects and to develop an instrument for use in future randomized trials. METHODS: We designed, translated and mailed a survey to 2,346 Japanese consumers of Sen-Sei-Ro self-designated as cancer patients. The survey assessed consumer demographics, cancer history, Sen-Sei-Ro consumption, and its perceived effects. We performed exploratory psychometric analyses to identify distinct, multi-item scales that could summarize perceptions of effects. RESULTS: We received completed questionnaires from 782 (33%) of the sampled Sen-Sei-Ro consumers with a cancer history. Respondents represented a broad range of cancer patients familiar with Sen-Sei-Ro. Nearly all had begun consumption after their cancer diagnosis. These consumers expressed consistently positive views, though not extremely so, with more benefit reported for more abstract benefits such as emotional and physical well-being than relief of specific symptoms. We identified two conceptually and empirically distinct and internally consistent summary scales measuring Sen-Sei-Ro consumers’ perceptions of its effects, Relief of Symptoms and Functional Well-being (Cronbach’s alpha: Relief of Symptoms, alpha =.74; Functional Well-Being, alpha =.91). CONCLUSIONS: Respondents to our survey of Sen-Sei-Ro consumers with cancer reported favorable perceived effects from its use. Our instrument, when further validated, may be a useful outcome in trials assessing this and other complementary and alternative medicine (CAM) substances in cancer patients.
PMID: 17967191 [PubMed - as supplied by publisher]

13. Therapy of myeloma in vivo using marine phospholipid in combination with Agaricus blazei Murill as an immune respond activator.
Murakawa K, Fukunaga K, Tanouchi M, Hosokawa M, Hossain Z, Takahashi K.Division of Marine Biosciences, Graduate School of Fisheries Sciences, Hokkaido University, Hokkaido, Japan.J Oleo Sci. 2007;56(4):179-88.
Mushroom (Agaricus blazei Murill) extract has been reported to possess antitumor effects through immune activation. Here, we investigated the beneficial effects of combining A. blazei extract with marine phospholipids in comparison to A. blazei extract alone on myeloma sp2 tumor suppression when orally administrated. The experimental groups designed for sp2 tumor bearing BALB/c nu/nu mice were drinks of: (1)control; (2)1.0 mg/mL squid phospholipid liposome alone; (3)0.5 mg/mL A. blazei Murill water extract alone; (4)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract in the form of those simple mixture; and (5)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract partially encapsulated. Orally administrated volumes amounted to approximately 5 mL per day per mouse for all groups. A. blazei Murill water extract alone and squid phospholipid alone served groups show moderate tumor suppression with total administrations of approximately 105 mg/mouse for squid phospholipid through out the experimental term. When both A. blazei Murill water extract and squid phospholipid were administrated simultaneously in a simple mixture form, promotional effect on cancer tumor suppression was observed. And when A. blazei Murill water extract was partially encapsulated in the squid phospholipid liposomes with total administrations being 105 mg/mouse for squid phospholipid, effect on cancer tumor suppression was more pronounced. Though there was no statistically significant difference in tumor sizes between the simple mixture form administrated group i.e. group (4) and the partially encapsulated form administrated group i.e. group (5), the tumor vanished mouse was seen in the partially encapsulated form administrated group. Thus it was concluded that combinational administration of the A. blazei Murill water extract and the marine phospholipid may be useful in myeloma sp2 therapy.
PMID: 17898480 [PubMed - indexed for MEDLINE]

14. Lack of carcinogenicity of lyophilized Agaricus blazei Murill in a F344 rat two year bioassay.
Lee IP, Kang BH, Roh JK, Kim JR.
Laboratory of Molecular Toxicology, Toxicological Research Center, Korea Food and Drug Administration, 5 Nokbun-Dong, Unpyong-Ku, Seoul 122-704, Republic of Korea.
Food Chem Toxicol. 2008 Jan;46(1):87-95. Epub 2007 Jul 13.
The Brazilian mushroom Agaricus blazei Murill has antimutagenic, antioxidant, immunostimulatory and antitumorigenic activities, and is increasingly consumed as a health food worldwide. We undertook the present study to evaluate the chronic toxicity and oncogenicity of A. blazei Murill in F344 rats. To establish a no-observed-adverse-effect level (NOAEL), four treatment groups of 100 rats each (50 males and 50 females) were fed a powder diet containing lyophilized A. blazei aqueous extract at 0, 6250, 12,500, and 25,000ppm for up to 2 years. During this period, there was no remarkable change in mean body weight, body weight gain, hematologic or serum chemistry parameters, or absolute or relative organ weights in control or treatment groups. Mortality in male treatment groups (26%, 16%, and 30%), however, was significantly lower than in controls (48%). Histopathological studies showed no increased incidence of tumors in any treatment group, and total tumor incidence across all groups was comparable to historical data. In conclusion, an A. blazei Murill lyophilized powder diet even at 25,000ppm (1176mg/kgb.w./day for male rats and 1518mg/kgb.w./day for female rats) resulted in no remarkable carcinogenic effects in F344 rats over a 2-year period. Therefore, the dietary NOAEL is 25,000ppm.
PMID: 17707568 [PubMed - in process]

15. Protective effects of beta-glucan extracted from Agaricus brasiliensis against chemically induced DNA damage in human lymphocytes.
Angeli JP, Ribeiro LR, Gonzaga ML, Soares Sde A, Ricardo MP, Tsuboy MS, Stidl R, Knasmueller S, Linhares RE, Mantovani MS.
Departamento de Biologia Geral, Universidade Estadual de Londrina, Londrina, PR, Brazil.
Cell Biol Toxicol. 2006 Jul;22(4):285-91. Epub 2006 Jun 26.
Beta-Glucans (BGs) are polysaccharides that are found in the cell walls of organisms such as bacteria, fungi, and some cereals. The objective of the present study was to investigate the genotoxic and antigenotoxic effects of BG extracted from the mushroom Agaricus brasiliensis (=Agaricus blazei Murrill ss. Heinemann). The mutagenic activity of BG was tested in single-cell gel electrophoresis assays with human peripheral lymphocytes. In addition, the protective effects against the cooked food mutagen 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) and (+/-)-anti-B[a]P-7,8-dihydrodiol-9,10-epoxide (BPDE), which is the main metabolite of B[a]P, and against ROS (H(2)O(2))-induced DNA damage, were studied. The results showed that the compound itself was devoid of mutagenic activity, and that a significant dose-dependent protective effect against damage induced by hydrogen peroxide and Trp-P-2 occurred in the dose range 20-80 microg/ml. To investigate the prevention of Trp-P-2-induced DNA damage, a binding assay was carried out to determine whether BG inactivates the amine via direct binding. Since no such interactions were observed, it is likely that BG interacts with enzymes involved in the metabolism of the amine.
PMID: 16802105 [PubMed - indexed for MEDLINE

16. Immunomodulatory effects of Agaricus blazei Murill in Balb/cByJ mice.
Chan Y, Chang T, Chan CH, Yeh YC, Chen CW, Shieh B, Li C.Department of Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan.
J Microbiol Immunol Infect. 2007 Jun;40(3):201-8.
BACKGROUND AND PURPOSE: Agaricus blazei Murill has been reported to possess biological effects that include immunomodulatory and tumoricidal activities, but its potential effects have not been systematically analyzed in vivo. We evaluated the immunomodulatory effects of A. blazei in Balb/cByJ mice. METHODS: 160 male Balb/cByJ mice were divided into four groups and treated with various quantities of intragastric A. blazei extract or distilled water for 8 to 10 weeks. Nine parameters, relating to general immune function or adaptive immunity against immunogen chicken oval albumin, were determined. RESULTS: The results revealed that mice receiving A. blazei extract exhibited significantly greater serum immunoglobulin G levels, increased T-cell numbers in spleen, and elevated phagocytic capability compared with controls. Consumption of A. blazei was also associated with significant increases in oval albumin-specific serum immunoglobulin G level, delayed-type hypersensitivity, splenocyte proliferation rate, and tumor necrosis factor-alpha secretion by splenocytes. CONCLUSIONS: Consumption of A. blazei extract was associated with significant enhancement of seven out of nine immune functions tested. We conclude that A. blazei Murill possesses a wide range of immunomodulatory effects in vivo.
PMID: 17639159 [PubMed - indexed for MEDLINE]

17. Interleukin-12- and interferon-gamma-mediated natural killer cell activation by Agaricus blazei Murill.
Yuminamochi E, Koike T, Takeda K, Horiuchi I, Okumura K.
Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan.
Immunology. 2007 Jun;121(2):197-206. Epub 2007 Mar 7.
Dried fruiting bodies of Agaricus blazei Murill (A. blazei) and its extracts have generally used as complementary and alternative medicines (CAMs). Here, we report that the oral administration of A. blazei augmented cytotoxicity of natural killer (NK) cells in wild-type (WT) C57BL/6, C3H/HeJ, and BALB/c mice. Augmented cytotoxicity was demonstrated by purified NK cells from treated wild-type (WT) and RAG-2-deficient mice, but not from interferon-gamma (IFN-gamma) deficient mice. NK cell activation and IFN-gamma production was also observed in vitro when dendritic cell (DC)-rich splenocytes of WT mice were coincubation with an extract of A. blazei. Both parameters were largely inhibited by neutralizing anti-interleukin-12 (IL-12) monoclonal antibody (mAb) and completely inhibited when anti-IL-12 mAb and anti-IL-18 mAb were used in combination. An aqueous extract of the hemicellulase-digested compound of A. blazei particle; (ABPC) induced IFN-gamma production more effectively, and this was completely inhibited by anti-IL-12 mAb alone. NK cell cytotoxicty was augmented with the same extracts, again in an IL-12 and IFN-gamma-dependent manner. These results clearly demonstrated that A. blazei and ABPC augmented NK cell activation through IL-12-mediated IFN-gamma production.
PMID: 17346284 [PubMed - indexed for MEDLINE]

18. Effects on gene expression and viral load of a medicinal extract from Agaricus blazei in patients with chronic hepatitis C infection.
Grinde B, Hetland G, Johnson E.
Division of Infectious Disease Control, Norwegian Institute of Public Health, Ullevål University Hospital, Oslo, Norway. bjorn.grinde@thi.no
Int Immunopharmacol. 2006 Aug;6(8):1311-4. Epub 2006 May 11.
Extracts from the mushroom Agaricus blazei Murill (AbM) are used extensively as a non-prescription remedy against cancer and infections, including hepatitis. We previously demonstrated a potent immunomodulating effect of a particular preparation on monocytes in vitro, and a protective effect on bacterial infections in mice. Here we report the effect on gene expression in peripheral blood cells from four chronic hepatitis C patients, using global (29 k) oligo-based, single channel microarrays. The viral load was slightly, but not significantly, decreased after 1 week of AbM treatment. The cytokine genes most strongly induced in vitro were not induced in vivo. The more notable changes in mRNA levels were related to genes involved in the G-protein coupled receptor signalling pathway, in cell cycling, and in transcriptional regulation. The results suggest that the beta-glucans of the extract, which presumably are responsible for cytokine induction, did not readily enter the blood, while other components, such as substances proposed to have anticancer effects, were active in the blood.
PMID: 16782544 [PubMed - indexed for MEDLINE]

20. Activation of antitumor immunity by intratumor injection of biological preparations [Article in Japanese]
Ebina T.
Division of Immunology, Miyagi Cancer Center Research Institute.
Gan To Kagaku Ryoho. 2003 Oct;30(11):1555-8.
The antitumor effects of biological response modifiers (BRMs) in an experimental mouse model using a double grafted tumor system were analyzed. Some BRMs prevented metastases by utilizing the anti-tumor immunological cascade reactions, which activate macrophages in the body. The following BRMs were analyzed: PSK was a hot water extract of cultured mycelia from Coliolus versicolor and a protein bound beta-glucan. Lentinan was purified from fruit bodies of Lentinus erodes and is a beta-glucan. The agaricus preparation was extracted from fruit bodies of Agaricus blazei and a protein-bound alpha-, beta-glucan. The M2 fraction was extracted from mycelia of Tricholoma matsutake and was a protein bound alpha-glucan. M1 fraction was purified from mycelia of T. matsutake and was an alpha-glucan. PSK cured both primary and metastatic tumors in the double grafted tumor system. Lentinan did not inhibit the growth of either primary or metastatic tumors. Agaricus preparation cured a primary tumor and inhibited the growth of a metastatic tumor. The M2 fraction prepared from Matsutake inhibited the growth of both primary and metastatic tumors. The M1 fraction did not inhibit either primary or metastatic tumors. Immunosuppressive acidic protein (IAP) is produced by activated macrophages. The PSK, Agaricus preparation and M2 fraction of the Matsutake preparation induced IAP but the lentinan and M1 fraction did not.
PMID: 14619462 [PubMed - indexed for MEDLINE]

21. Anti-genotoxic effect of aqueous extracts of sun mushroom (Agaricus blazei Murill lineage 99/26) in mammalian cells in vitro.
Martins de Oliveira J, Jordao BQ, Ribeiro LR, Ferreira da Eira A, Mantovani MS.
Departamento de Biologia Geral CCB, Universidade Estadual de Londrina, Campus Universitario, Cx Postal 6001, Londrina, PR, Brazil.
Food Chem Toxicol. 2002 Dec;40(12):1775-80.
The “sun mushroom” is the popular name for the Agaricus blazei Murill fungus, a mushroom native to south-eastern Brazil, which has been frequently used in popular medicine mainly in the form of tea to treat various ailments (stress, diabetes, etc.). In the present study, the genotoxic and/or anti-genotoxic effects ofA. blazei on mammalian cells in culture was assessed by checking the increase or reduction of micronucleus (MN) frequency and comets. The sun mushroom (lineage 99/26) was used as aqueous extracts prepared (2.5%) at three different temperatures (60, 25 and 4 degrees C). The in vitro micronucleus (MN) test in binucleated cells and comet assay were used in V79 cells cultivated in HAM-F10+DMEM medium (1:1), supplemented with 10% of fetal bovine serum. The experiments were divided into four treatment types: 1. Negative control; 2. Positive control with MMS; 3. Treatments with the three forms of extracts (60, 25 and 4 degrees C); and 4. Treatments with the extracts in different associations (simultaneous, pre-treatment, post-treatment and simultaneous after pre-incubation for 1 h) with MMS. None of the A. blazei extracts show genotoxic activity. In the comet assay no protecting effect was found. The results obtained in the MN test showed that the three forms of extracts used had protective activity, suggesting that the compound or active ingredients of A. blazei are always present in these extracts. The greater protective efficiency of the simultaneous treatment and simultaneous treatment with pre-incubation mixture with MMS suggests that the extracts have an antimutagenic action of the desmutagenic type.
PMID: 12419691 [PubMed - indexed for MEDLINE]

22. Antimutagenic effect of Agaricus blazei Murrill mushroom on the genotoxicity induced by cyclophosphamide.
Delmanto RD, de Lima PL, Sugui MM, da Eira AF, Salvadori DM, Speit G, Ribeiro LR.Departamento de
Patologia, Faculdade de Medicina, UNESP, 18618-000, SP, Botucatu, Brazil.
Mutat Res. 2001 Sep 20;496(1-2):15-21.
Agaricus blazei Murrill extracts have previously been shown to have anticarcinogenic and antimutagenic properties. These results suggest that antimutagenic activity, besides the modulation of the immune system, might be involved in the anticarcinogenic action of A. blazei. To investigate the possible antimutagenic effect of A. blazei in vivo, we evaluated its effect on clastogenicity induced by cyclophosphamide (CP) in mice, using the micronucleus test in bone marrow (MNPCE) and in peripheral blood (MNRET). Male Swiss mice were treated with CP (25 or 50mg/kg i.p.) or with CP plus mushroom solution at three different temperatures: 4, 21, and 60 degrees C. Aqueous solution of a mixture from various lineages of the mushroom inhibited induction of micronuclei by CP in bone marrow and in peripheral blood of mice. In contrast to the mixture of lineages, a single isolated lineage did not lead to a reduction of CP-induced MN frequencies in either bone marrow or blood cells of mice. The results suggest that under certain circumstances these mushrooms exhibit antimutagenic activities that might contribute to an anticarcinogenic effect.
PMID: 11551476 [PubMed - indexed for MEDLINE]

23. Anti-tumor polysaccharide from the mycelium of liquid-cultured Agaricus blazei mill.
Mizuno M, Minato K, Ito H, Kawade M, Terai H, Tsuchida H.
Graduate School of Science and Technology, Kobe University, Japan.
Biochem Mol Biol Int. 1999 Apr;47(4):707-14.
Anti-tumor active polysaccharide against Sarcoma 180 was isolated by DEAE-Sepharose CL-6B and Sepharose 4B column chromatography from the hot-water soluble fraction of the mycelium of liquid-cultured Agaricus blazei mill. This polysaccharide did not react with antibodies of anti-tumor polysaccharides such as lentinan, gliforan, and FIII-2-b which is one of anti-tumor polysaccharides from Agaricus blazei. Moreover, the analyses of 13C-NMR and GC-MS suggested that this polysaccharide was preliminarily glucomannan with a main chain of beta-1,2-linked D-mannopyranosyl residues and beta-D-glucopyranosyl-3-O-beta-D-glucopyranosyl residues as a side chain. This polysaccharide was completely different from the anti-tumor polysaccharide from fruiting body of Agaricus blazei, beta-1,6-glucan.
PMID: 10319424 [PubMed - indexed for MEDLINE]

24. Antitumor beta glucan from the cultured fruit body of Agaricus blazei.
Ohno N, Furukawa M, Miura NN, Adachi Y, Motoi M, Yadomae T.
Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy
& Life Science,
Hachioji, Japan. ohnonao@ps.toyaku.ac.jp
Biol Pharm Bull. 2001 Jul;24(7):820-8.
Agaricus blazei is a medically important mushroom widely eaten and prescribed in Japan. Polysaccharide fractions were prepared from cultured A. blazei by repeated extraction with hot water (AgHWE), cold NaOH (AgCA), and then hot NaOH (AgHA). By chemical, enzymic, and NMR analyses, the primary structures of AgHWE, AgCA, and AgHA were mainly composed of 1,6-beta-glucan. Among these fractions, the NaOH extracts showed antitumor activity against the solid form of Sarcoma 180 in ICR mice. To demonstrate the active component in these fractions, several chemical and enzymic treatments were applied. These fractions were found to be i) neutral beta-glucan passing DEAE-Sephadex A-25, ii) resistant to periodate oxidation (I/B) and subsequent partial acid hydrolysis (I/B/H), iii) resistant to a 1,3-beta-glucanase, zymolyase, before I/B, but sensitive after I/B/H. In addition, after I/B/H treatment of the neutral fraction of AgCAE, a signal around 86 ppm attributable to 1,3-beta glucosidic linkage was detectable in the 13C-NMR spectrum. These facts strongly suggest that a highly branched 1,3-beta-glucan segment forms the active center of the antitumor activity.
PMID: 11456124 [PubMed - indexed for MEDLINE]

25. Antitumor effect of a peptide-glucan preparation extracted from Agaricus blazei in a double-grafted tumor system in mice.
Ebina T, Fujimiya Y.
Division of Immunology, Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan.
Biotherapy. 1998;11(4):259-65.
The antitumor effect of extracts obtained from the fruit body of Agaricus blazei Murill was examined in a double-grafted tumor system, in which BALB/c mice received simultaneous intradermal injections of Meth-A tumor cells in both the right (10(6) cells) and left flank (2 x 10(5) cells), and were then injected with 5 mg of extracts of A. blazei in the right tumor on days 3, 4 and 5. Intratumoral administration of ethanol-soluble (Fraction 1), water-ethanol-soluble (Fraction 2), ammonium oxalate-soluble (Fraction 3) and ammonium oxalate-insoluble (Fraction 4) fractions resulted in inhibition of tumor growth, with Fraction 3 showing the most tumoricidal activity, producing regression of the right tumor and inhibition of growth of the left, non-injected tumor. The maximum effect was obtained using 0.5 mg of Fraction 3 and this amount was used in subsequent experiments. The antitumor effect of intratumorally administered Fraction 3 was enhanced by oral ad lib administration of feed containing 0.083% of Fraction 3. When immunized spleen cells from mice that had been cured by intratumoral administration of 0.5 mg of Fraction 3 were directly injected (2 x 10(7) cells/mouse) into the Meth-A tumor, tumor growth was inhibited. The tumor cells on day 7 from the Fraction 3-treated right tumor and from the left tumor were cultured for 24 h and their culture supernatants were assayed for neutrophil or macrophage chemotactic activity. Significant macrophage chemotactic factor activity was detected in the culture media from the left tumor tissue. Serum levels of immunosuppressive acidic protein (IAP), produced by activated macrophages and neutrophils, increased transiently soon after intradermal injection of 0.5 mg of Fraction 3. These results suggest that regression of the left non-injected tumor was due to an immune reaction, involving induction of cytotoxic cells in the spleen, and the release of chemotactic factors in the distant tumor.
PMID: 9950102 [PubMed - indexed for MEDLINE]

26. Isolation of an Antitumor Compound from Agaricus blazei Murill and Its Mechanism of Action1
Takeshi Takaku*, Yoshiyuki Kimura2 and Hiromichi Okuda
(Journal of Nutrition. 2001;131:1409-1413.) © 2001 The American Society for Nutritional Sciences
Second Department of Medical Biochemistry and *Central Research Laboratory, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-0295, Japan
Yao Xue Xue Bao. 1997 Jun;32(6):431-7.
The Basidiomycete fungus Agaricus blazei Murill has traditionally been used as a health food for the prevention of cancer, diabetes, hyperlipidemia, arteriosclerosis and chronic hepatitis. In the present study, we examined the antitumor activities of various substances isolated from the lipid fraction of A. blazei. Tumor growth was retarded by the oral administration of the lipid fraction extracted from A. blazei with a chloroform/methanol mixture in sarcoma 180–bearing mice. The substance with the antitumor activity in the lipid fraction was isolated via silica gel column chromatography, eluted with an acetonitrile/methanol (3:2) mixture and identified as ergosterol by direct comparison of the 1H NMR and mass spectrometry spectral data of an authentic sample. The oral administration of ergosterol to sarcoma 180–bearing mice significantly reduced tumor growth at doses of 400 and 800 mg/kg administered for 20 d without side effects, such as the decreases in body, epididymal adipose tissue, thymus, and spleen weights and leukocyte numbers induced by cancer chemotherapy drugs. Ergosterol had no cytotoxicity against tumor cells. To clarify the antitumor activity of ergosterol, we examined the effects of ergosterol on tumor-induced angiogenesis using two in vivo models. Intraperitoneal administration of ergosterol at doses of 5, 10 and 20 mg/kg for 5 consecutive d inhibited the neovascularization induced by Lewis lung carcinoma cell–packed chambers, suggesting that either ergosterol or its metabolites may be involved in the inhibition of tumor-induced neovascularization. Therefore, we further examined the inhibitory effects of ergosterol on Matrigel-induced neovascularization. Female C57BL/6 mice were subcutaneously inoculated with Matrigel containing acidic fibroblast growth factor and heparin with or without ergosterol. Ergosterol inhibited the Matrigel-induced neovascularization, suggesting that ergosterol directly inhibits Matrigel-induced neovascularization. From these results, it seems likely that the antitumor activity of ergosterol might be due to direct inhibition of angiogenesis induced by solid tumors. This is the first report of ergosterol as an antiangiogenic substance.

27. Antimutagenic and bactericidal substances in the fruit body of a Basidiomycete Agaricus blazei, Jun-17 [Article in Japanese]
Osaki Y, Kato T, Yamamoto K, Okubo J, Miyazaki T.
Tokyo College of Pharmacy, Japan.
Yakugaku Zasshi. 1994 May;114(5):342-50.
The fruit body of a Basidiomycete Agaricus blazei, Jun-17 (Himematsutake) was extracted with hexane and chloroform-methanol (2:1, v/v), and the antimutagenic effect of the extracts was examined using an Ames/Salmonella/microsome assay. Both extracts of Agaricus inhibited the mutagenicity of benzo[a]pyrene(B[a]P). The hexane extract was purified by silica gel column chromatography and high performance liquid chromatography (HPLC), and linoleic acid was isolated as a main substance having antimutagenic activity. Fr. IIa, IIb, IIc and IIb, which reduced the number of His+ revertant colonies induced by B[a]P, were separated from the chloroform-methanol extract by silica gel column chromatography and HPLC. An antimutagenic substance in Fr. IIa was linoleic acid. From Fr. IIb, a bactericidal, not antimutagenic, substance was isolated and identified as 13-hydroxy cis-9, trans-11-octadecadienoic acid (13ZE-LOH). Antimutagenic substances in Fr. IIc and IId were not purified. The possible source and mechanism of formation of 13ZE-LOH are discussed.
PMID: 8014843 [PubMed - indexed for MEDLINE]

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