Cryptococcus neoformans and Cryptococcus gattii are found in distinct environments with some overlap around different parts of the world. However, no systematic surveys of these two pathogens have been reported from Puerto Rico, a tropical island uniquely situated between mainland USA and countries in South America. We carried out an exhaustive environmental survey in southwestern Puerto Rico for pathogenic Cryptococcus species. Twenty-two presumptive isolates of C. gattii from cacti and tree detritus were characterized in detail by physiological and molecular methods and seventeen strains were confirmed as C. gattii. Cryptococcus gattii isolates were haploid and majority of them were MATa [corrected] strains. Sixteen out of seventeen C. gattii isolates belonged to VGII/AFLP6 genotype while one isolate was a VGIV/AFLP7 genotype. The results are significant as Puerto Rico strains are distinct from VGIII/AFLP5 strains reported from Southern California, but similar to C. gattii VGII/AFLP6 molecular type implicated in recent outbreaks of cryptococcosis in Pacific Northwest and British Columbia, Canada, but different in its M13 fingerprinting, and a common genotype in South America.

For the past several decades, cancer patients in the U.S. have chosen the use of natural products as an alternative or complimentary medicine approach to treat or improve their quality of life via reduction or prevention of the side effects during or after cancer treatment. The genus Ganoderma includes about 80 species of mushrooms, of which several have been used for centuries in traditional Asian medicine for their medicinal properties, including anticancer and immunoregulatory effects. Numerous bioactive compounds seem to be responsible for their healing effects. Among the approximately 400 compounds produced by Ganoderma spp., triterpenes, peptidoglycans and polysaccharides are the major physiologically-active constituents. Ganoderma anticancer effects are attributed to its efficacy in reducing cancer cell survival and growth, as well as by its chemosensitizing role. In vitro and in vivo studies have been conducted in various cancer cells and animal models; however, in this review, we focus on Ganoderma's efficacy on breast cancers. Evidence shows that some species of Ganoderma have great potential as a natural therapeutic for breast cancer. Nevertheless, further studies are needed to investigate their potential in the clinical setting and to translate our basic scientific findings into therapeutic interventions for cancer patients.

contributed equally to this work.Abstract, p.

Inflammatory Breast Cancer (IBC) is an aggressive and lethal form of breast cancer with symptoms that include a diffuse redness and swelling of the breast. These signs resemble an inflammation, which may suggest the involvement of both immunological and inflammatory processes in the pathogenesis of this disease. Unique pathological findings indicate the presence of tumor emboli invading the dermal lymphatics of the breast; a process considered to be responsible for the inflammatory phenotype. However, these tumors produce inflammatory mediators such interleukin-6 (IL-6), which may act as a growth factor to contribute to cancer progression. Thus, cancer cells stimulated with or that secrete IL-6 via an autocrine loop, auto-activate these pathways resulting in malignant features such as enhanced invasion and increased metastasis. Our published data demonstrates that the medicinal mushroom, Ganoderma lucidum (Reishi), disintegrates tumor emboli, and reduces tumor growth. We hypothesize that IL-6 enhances while Reishi reduces IBC progression via modulation of the JAK-2/STAT-3 pathway both in vitro and in vivo. SUM-149 wound-healing assays were performed to determine the migratory phenotype of IBC cells stimulated with IL-6 or treated with Reishi. Immunoblots of SUM-149 and KPL-4 IBC cell lysates were performed to detect the expression of the JAK-2/STAT-3 signaling pathway and its downstream effectors upon Reishi treatment. Cell fractioning was performed to delineate if the proteins affected by Reishi belong to the nuclear or cytoplasmic fraction of the cell. For in vivo studies, severe combined immunodeficient (SCID) mice were injected with SUM-149 IBC cells in their mammary fat pad and treated via oral gavage with various concentrations of Reishi for 10wk. Our data from the in vitro studies indicate that IL-6 causes wound closure after 24h, an effect prevented by Reishi. Additionally, our data suggests that Reishi has an immunomodulatory role, demonstrated by its ability to reduce the phosphorylation of the IL-6/JAK-2/STAT-3 pathway. Cell fractioning of SUM-149 and KPL-4 cells show a drastic reduction in the expression of proteins corresponding to the nuclear fraction of the cell when treated Reishi. Finally, our in vivo data show a dose dependent reduction by 3.0 and 5.5 fold in tumor volume, when mice where treated with 7mg/kg BW and 14mg/kg BW of Reishi, respectively. This data suggests that Reishi may be used as a targeted therapeutic for women afflicted with IBC, for whom no direct therapeutics are currently available. We thank Dr. Kurebayashi (Kawasaki Medical School, Japan) for providing KPL-4 cells. This project was sponsored by Title V PPOHA US Department of Education #P031M105050 to UCC, NIH/RCMI #G12 MD007583 to UCC, NIH/INBRE #P20 GM103475 to UPR/UCC and a research donation from the Commonwealth of Puerto Rico to UCC's University Center of Integral and Complementary Medicine (CUMIC)/MMM. Citation Format: Yaliz Loperena-Alvarez, Luis A. Cubano, Michelle M. Martínez-Montemayor. Role of IL-6 in inflammatory breast cancer and its modulation by Ganoderma lucid (Reishi). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3673. doi:10.1158/1538-7445.AM2014-3673

Ganoderma lucidum (Reishi) is a medicinal mushroom used to treat among many diseases, hyperglycemia, inflammation and cancer. Our laboratory studies the effects Reishi, as an alternative approach to treat Inflammatory Breast Cancer (IBC). IBC is an aggressive form of breast cancer. IBC symptoms include a diffuse redness and swelling of the breast, which resembles inflammation and may suggest the involvement of both immunological and inflammatory processes in the pathogenesis of this disease. Pathological findings indicate the presence of tumor emboli invading the dermal lymphatics of the breast. In these tumors pro-inflammatory mediators such as interleukin-8 (IL-8), IL-6, and Monocyte Chemotactic Protein 1 (MCP-1) are produced to contribute to cancer progression. These proteins are regulators of the immune system but in the tumor microenvironment they can act as growth factors, allowing tumor cell survival, invasion and metastasis. Our published data demonstrates that Reishi disintegrates tumor emboli, reduces tumor growth and modulates the abundance of plasma membrane proteins involved in downstream signaling of invasion cascades. Herein, we are characterizing cytokine production in vitro and in vivo and their modulation upon Reishi treatment. SUM-149 conditioned media was analyzed via cytokine bead array to quantify cytokine production after 0 or 0.5mg/mL Reishi treatment at different time points. For in vivo studies, severe combined immunodeficient (SCID) mice were treated daily for two weeks with 0, 7, 14 or 28mg/kg BW of Reishi via oral gavage. At two weeks the mice were injected with SUM-149 IBC cells in their mammary fat pad and continued oral gavage treatment for an additional 14 weeks. Our in vitro study data shows that SUM-149 cells secrete IL-6 and IL-8 and that secretion of these cytokines, which are known to behave as growth factors, was decreased by 23% and 76% respectively after 48 hours of Reishi treatment. As of 10 weeks of the study, tumor size is reduced in all Reishi treated animals when compared with vehicles. Interestingly, Reishi shows a tendency to decrease the expression of MCP-1 and IL-6 in plasma, while a slight increase in TNF-α is observed. Our results show that Reishi has an immunomodulatory role, demonstrated by its ability to reduce cytokine production both in cells and in mice plasma. Reishi may be used as a targeted therapeutic for women afflicted with IBC, for whom no direct therapeutics are currently available. This project was sponsored by Title V PPOHA US Dept of Education #P031M105050 to UCC, NIH/RCMI #2G12RR003035 to UCC, NIH/INBRE #5P20RR016470 to UPR/UCC, NIH/NCRR #U54RR026139 to UPR/UCC, and a research donation from the Commonwealth of Puerto Rico to UCC-Centro Universitario de Medicina Integral y Complementaria (CUMIC) / MMM. Citation Format: Yaliz Loperena-Alvarez, Luis A. Cubano, Michelle M. Martínez-Montemayor. Effects of Ganoderma lucidum on cytokine expression in Inflammatory Breast Cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4727. doi:10.1158/1538-7445.AM2013-4727