Plants against cancer: the immune-boosting herbal microbiome: not of the plant, but in the plant. Basic concepts, introduction, and future resource for vaccine adjuvant discovery.

The presence of microorganism communities (MOCs) comprised of bacteria, fungi, archaea, algae, protozoa, viruses, and the like, are ubiquitous in all living tissue, including plant and animal. MOCs play a significant role in establishing innate and acquired immunity, thereby influencing susceptibility and resistance to disease. This understanding has fostered substantial advancements in several fields such as agriculture, food science/safety, and the development of vaccines/adjuvants, which rely on administering inactivated-attenuated MOC pathogens. Historical evidence dating back to the 1800s, including reports by Drs Busch, Coley, and Fehleisen, suggested that acute febrile infection in response to "specific microbes" could trigger spontaneous tumor remission in humans. This discovery led to the purposeful administration of the same attenuated strains, known as "Coley's toxin," marking the onset of the first microbial (pathogen) associated molecular pattern (MAMPs or PAMPs)-based tumor immunotherapy, used clinically for over four decades. Today, these same MAMPS are consumed orally by billions of consumers around the globe, through "specific" mediums (immune boosting "herbal supplements") as carriers of highly concentrated MOCs accrued in roots, barks, hulls, sea algae, and seeds. The American Herbal Products Association (AHPA) mandates microbial reduction in botanical product processing but does not necessitate the removal of dead MAMP laden microbial debris, which we ingest. Moreover, while existing research has focused on the immune-modulating role of plant phytochemicals, the actual immune-boosting properties might instead reside solely in the plant's MOC MAMP laden biomass. This assertion is logical, considering that antigenic immune-provoking epitopes, not phytochemicals, are known to stimulate immune response. This review explores a neglected area of research regarding the immune-boosting effects of the herbal microbiome - a presence which is indirectly corroborated by various peripheral fields of study and poses a fundamental question: Given that food safety focuses on the elimination of harmful pathogens and crop science acknowledges the existence of plant microbiomes, what precisely are the immune effects of ingesting MAMPs of diverse structural composition and concentration, and where are these distributed in our botanicals? We will discuss the topic of concentrated edible MAMPs as acid and thermally stable motifs found in specific herbs and how these would activate cognate pattern recognition receptors (PPRs) in the upper gut-associated lymphoid tissue (GALT), including Peyer's patches and the lamina propria, to boost antibody titers, CD8+ and CD4+ T cells, NK activity, hematopoiesis, and facilitating M2 to M1 macrophage phenotype transition in a similar manner as vaccines. This new knowledge could pave the way for developing bioreactor-grown/heat-inactivated MOC therapies to boost human immunity against infections and improve tumor surveillance.

Therapeutic Potential of Thymoquinone in Triple-Negative Breast Cancer Prevention and Progression through the Modulation of the Tumor Microenvironment.

To date, the tumor microenvironment (TME) has gained considerable attention in various areas of cancer research due to its role in driving a loss of immune surveillance and enabling rapid advanced tumor development and progression. The TME plays an integral role in driving advanced aggressive breast cancers, including triple-negative breast cancer (TNBC), a pivotal mediator for tumor cells to communicate with the surrounding cells via lymphatic and circulatory systems. Furthermore, the TME plays a significant role in all steps and stages of carcinogenesis by promoting and stimulating uncontrolled cell proliferation and protecting tumor cells from the immune system. Various cellular components of the TME work together to drive cancer processes, some of which include tumor-associated adipocytes, fibroblasts, macrophages, and neutrophils which sustain perpetual amplification and release of pro-inflammatory molecules such as cytokines. Thymoquinone (TQ), a natural chemical component from black cumin seed, is widely used traditionally and now in clinical trials for the treatment/prevention of multiple types of cancer, showing a potential to mitigate components of TME at various stages by various pathways. In this review, we focus on the role of TME in TNBC cancer progression and the effect of TQ on the TME, emphasizing their anticipated role in the prevention and treatment of TNBC. It was concluded from this review that the multiple components of the TME serve as a critical part of TNBC tumor promotion and stimulation of uncontrolled cell proliferation. Meanwhile, TQ could be a crucial compound in the prevention and progression of TNBC therapy through the modulation of the TME.

Milk thistle seed extract protects rat C6 astroglial cells from acute cocaine toxicity.

Cocaine is a powerful addictive drug, widely abused in most Western countries. It easily reaches various domains within and outside of the central nervous system (CNS), and triggers varying levels of cellular toxicity. No pharmacological treatment is available to alleviate cocaine-induced toxicity in the cells without side-effects. Here, we discerned the role of milk thistle (MT) seed extract against cocaine toxicity. First, we investigated acute cytotoxicity induced by treatment with 2, 3 and 4 mM cocaine for 1 h in astroglial, liver and kidney cells in vitro, and then in living shrimp larvae in vivo. We showed that astroglial cells are more sensitive to cocaine than liver, kidney cells or larvae. Cocaine exposure disrupted the general architecture of astroglial cells, induced vacuolation, decreased cell viability, and depleted the glutathione (GSH) level. These changes may represent the underlying pathology of cocaine in the astrocytes. By contrast, MT pretreatment (200 µg/ml) for 30 min sustained the cell morphological features and increased both cell viability and the GSH level. Besides its protective effects, the MT extract was revealed to be non-toxic to astroglial cells, and displayed high free-radical scavenging activity. The results from this study suggest that enhanced GSH level underlies cell protection, and indicate that compounds that promote GSH synthesis in the cells may be beneficial against cocaine toxicity.

The attenuation of early benzo(a)pyrene-induced carcinogenic insults by diallyl disulfide (DADS) in MCF-10A cells.

Diallyl disulfide (DADS), a garlic organosulfur compound, has been researched as a cancer prevention agent; however, the role of DADS in the suppression of cancer initiation in nonneoplastic cells has not been elucidated. To evaluate DADS inhibition of early carcinogenic events, MCF-10A cells were pretreated (PreTx) with DADS followed by the ubiquitous carcinogen benzo(a)pyrene (BaP), or cotreated (CoTx) with DADS and BaP for up to 24 h. The cells were evaluated for changes in cell viability/proliferation, cell cycle, induction of peroxide formation, and DNA damage. BaP induced a statistically significant increase in cell proliferation at 6 h, which was attenuated with DADS CoTx. PreTx with 6 and 60 µM of DADS inhibited BaP-induced G2/M arrest by 68% and 78%, respectively. DADS, regardless of concentration or method, inhibited BaP-induced extracellular aqueous peroxide formation within 24 h. DADS attenuated BaP-induced DNA single-strand breaks at all time points through both DADS Pre- and CoTx, with significant inhibition for all treatments sustained after 6 h. DADS was effective in inhibiting BaP-induced cell proliferation, cell cycle transitions, reactive oxygen species, and DNA damage in a normal cell line, and thus may inhibit environmentally induced breast cancer initiation.

Regulation of rat MOR-1 gene expression after chronic intracerebroventricular administration of morphine.

The µ-opioid receptor is the primary site for the action of morphine. In the present study, we investigated the regulation of the µ-opioid receptor mRNA levels in the locus ceruleus, ventral tegmental area, nucleus accumbens and hypothalamus of the rat brain following intracerebroventricular administration of morphine for 7 days. The isolated mRNA from these regions was subjected to real-time quantitative RT-PCR to determine the regulation of µ-opioid receptor gene expression. It was observed that 7 days of treatment with morphine significantly down-regulated the µ-opioid receptor mRNA levels in the hypothalamus of the brain in comparison to the control group. However, the µ-opioid receptor levels in the locus ceruleus, ventral tegmental area and nucleus accumbens regions remained the same as the control levels. Down-regulation of µ-opioid receptor mRNA levels in the hypothalamus region of the brain indicates the probable role of opioids to influence neuroendocrine function. The results further indicate that cellular adaptation for morphine tolerance is tissue-specific. These findings help us to understand the mechanism of morphine tolerance in various regions of the brain.

Differential cytotoxicity of triphala and its phenolic constituent gallic acid on human prostate cancer LNCap and normal cells.

BACKGROUND: Prostate cancer is one of the most commonly diagnosed solid malignancies among US men. We identified gallic acid (GA) as a major bioactive cytotoxic constituent of a polyherbal Ayurvedic formulation - triphala (TPL). Both TPL and GA were evaluated on (AR)(+) LNCaP prostate cancer and normal epithelial cells. MATERIALS AND METHODS: Total polyphenols in TPL were determined using Folin and Ciocalteu method, followed by GA quantitation by high performance liquid chromatography. Cell toxicity was evaluated by crystal violet after 24, 48, 72 and 96 h. RESULTS: TPL contains 40% unidentified polyphenolic acids, of which 2.4% comprised GA. GA induced severe morphological alterations and was about 3-fold more cytotoxic towards cancer cells than TPL. This activity increased further in the presence of dihydrotestosterone. GA toxicity on normal cells was low at 72 h. Combination of GA with flutamide caused higher toxicity to cancer cells than either of the compounds alone. CONCLUSION: GA appears to have promising anticancer activity.

Potent cytotoxic activity of Saururus cernuus extract on human colon and breast carcinoma cultures under normoxic conditions.

BACKGROUND: Saururus cernuus (Sc) is a small plant, used for the treatment of various human inflammations. The present study aimed at investigating the cytotoxic potential of the methanolic extract of this plant against brine shrimp larvae and human carcinoma cells at normoxic conditions. MATERIALS AND METHODS: The in vivo lethality test was evaluated at various doses against brine shrimp larvae at different time periods. Similarly, the extract was tested for 48 h at various concentrations against human CL-18 and MDA-MB-231 carcinoma cell lines and the toxicity was evaluated using the dye binding crystal-violet assay method. RESULTS: In the shrimp assay, the extract was very active, with ED50 values ranging from 1.83 +/- 0.2 to 2.79 +/- 0.2 microg/ml at various incubation periods. The extract was also very potent in human CL-18 and MDA-MB-231 cultures with LD50 values of 1.9 +/- 0.17 and 0.26 +/- 0.03 microg/ml, respectively. CONCLUSION: The results of this study indicate that Sc extract contains very stable, potent anticancer compounds, which gain access into the cells quickly and kill carcinoma cells and shrimp larvae at normoxic conditions.

Licamichauxiioic-A and -B acids--two ent-kaurene diterpenoids from Licania michauxii.

Bioassay-guided fractionation allowed the isolation of two new cytotoxic ent-kaurene diterpenoids, licamichauxiioic-A and -B acids (1 and 2) from the root extract of Licania michauxii Prance (Chrysobalanaceae). They were characterized as ent-15-oxo-9(11),16-kauradien-19-oic acid (1) and ent-15-oxo-13(14),16-kauradien-19-oic acid (2) by various spectroscopic methods, in particular, 1D and 2D NMR spectra, and chemical evidence.