Inhibition of Gli/hedgehog signaling in prostate cancer cells by "cancer bush" Sutherlandia frutescens extract.

Sutherlandia frutescens is a medicinal plant, traditionally used to treat various types of human diseases, including cancer. Previous studies of several botanicals link suppression of prostate cancer growth with inhibition of the Gli/hedgehog (Gli/Hh) signaling pathway. Here we hypothesized the anti-cancer effect of S. frutescens was linked to its inhibition of the Gli/Hh signaling in prostate cancer. We found a dose- and time-dependent growth inhibition in human prostate cancer cells, PC3 and LNCaP, and mouse prostate cancer cell, TRAMP-C2, treated with S. frutescens methanol extract (SLE). We also observed a dose-dependent inhibition of the Gli-reporter activity in Shh Light II and TRAMP-C2QGli cells treated with SLE. In addition, SLE can inhibit Gli/Hh signaling by blocking Gli1 and Ptched1 gene expression in the presence of a Gli/Hh signaling agonist (SAG). A diet supplemented with S. frutescens suppressed the formation of poorly differentiated carcinoma in prostates of TRAMP mice. Finally, we found Sutherlandioside D was the most potent compound in the crude extract that could suppress Gli-reporter in Shh Light II cells. Together, this suggests that the S. frutescens extract may exert anti-cancer effect by targeting Gli/Hh signaling, and Sutherlandioside D is one of the active compounds.

Evaluation of visible diffuse reflectance spectroscopy in liver tissue: validation of tissue saturations using extracorporeal circulation.

SIGNIFICANCE: Real-time information about oxygen delivery to the hepatic graft is important to direct care and diagnose vascular compromise in the immediate post-transplant period. AIM: The current study was designed to determine the utility of visible diffuse reflectance spectroscopy (vis-DRS) for measuring liver tissue saturation in vivo. APPROACH: A custom-built vis-DRS probe was calibrated using phantoms with hemoglobin (Hb) and polystyrene microspheres. Ex vivo (extracorporeal circulation) and in vivo protocols were used in a swine model (n = 15) with validation via blood gas analysis. RESULTS: In vivo absorption and scattering measured by vis-DRS with and without biliverdin correction correlated closely between analyses. Lin's concordance correlation coefficients are 0.991 for µa and 0.959 for µs ' . Hb measured by blood test and vis-DRS with (R2 = 0.81) and without (R2 = 0.85) biliverdin correction were compared. Vis-DRS data obtained from the ex vivo protocol plotted against the PO2 derived from blood gas analysis showed a good fit for a Hill coefficient of 1.67 and P50 = 34 mmHg (R2 = 0.81). A conversion formula was developed to account for the systematic deviation, which resulted in a goodness-of-fit R2 = 0.76 with the expected oxygen dissociation curve. CONCLUSIONS: We show that vis-DRS allows for real-time measurement of liver tissue saturation, an indicator for liver perfusion and oxygen delivery.

27-Hydroxycholesterol Is an Estrogen Receptor ß-Selective Negative Allosteric Modifier of 17ß-Estradiol Binding.

Estrogens bind to two nuclear estrogen receptor (ER) subtypes, ERα and ERß, which are expressed in differing amounts in various tissues. The endogenous estrogen, 17ß-estradiol (E2), binds to both subtypes with nearly equal affinity and is the prototypical agonist. Selective estrogen receptor modulators (SERMs) may bind to both subtypes with equivalent affinities but have agonist activities in some tissues while having antagonist activities in others. In the present study, we demonstrate that the first reported endogenous SERM, 27-hydroxycholesterol (27-OHC), binds preferentially (>100-fold) to ERß over ERα. Furthermore, 27-OHC is not able to fully compete with E2 binding, suggesting the two may bind at different sites. We provide an allosteric ternary complex model for the simultaneous binding of 27-OHC and E2 to ERß, which accurately describes the binding data we have observed. We conclude that 27-OHC is a negative allosteric modifier of E2 binding, with an inhibitor constantof 50 nM and cooperativity factor (α) of 0.036. We also propose an in silico three-dimensional model of the simultaneous binding to guide future experiments. Further study of this unique binding model may allow for the discovery of novel ERß-selective ligands and potentially explain the lack of effectiveness of ERß-selective agonists in humans vs preclinical models.

Mastoid and Inner Ear Measurements in Patients With Menière's Disease.

OBJECTIVE: To determine the relationship between radiographic temporal bone anatomy of patients with Menière's disease in medically and surgically managed populations versus controls. STUDY DESIGN: Retrospective chart review. SETTING: Two tertiary referral centers. PATIENTS: Adults older than 18 years with Menière's disease treated with endolymphatic sac decompression (ESD) or medical management (non-ESD) versus controls. INTERVENTIONS: Magnetic resonance imaging and computed tomography imaging studies of the temporal bones were reviewed by blinded radiologists. MAIN OUTCOME MEASURES: Radiographic temporal bone dimensions were measured in Menière's disease and control patients. Age, sex, symptoms, audiogram data, academy classification of Menière's disease, and follow-up were recorded. Statistical analysis was performed to compare outcome measures across groups and demographics. RESULTS: A total of 90 imaging studies were reviewed (ESD = 22; non-ESD = 30; control = 38). ESD and non-ESD groups had similar pure-tone averages (33.9 ±â€Š20.6 versus 41.6 ±â€Š22.6 dB HL; p = 0.21) and frequency of definite Menière's disease (59.1% versus 53.3%; p = 0.68). There was no significant trend between groups for any measurement. One nonsignificant trend existed in mean vestibule length, increasing from the control (5.45 ±â€Š0.54 mm), non-ESD (5.80 ±â€Š0.97 mm), and ESD (5.94 ±â€Š0.81 mm) group. In a combined Menière's group, mean vestibule length was significantly greater than controls (5.86 ±â€Š0.89 versus 5.45 ±â€Š0.54 mm; p = 0.008) and mean vestibule width significantly less (2.99 ±â€Š0.46 versus 3.19 ±â€Š0.39 mm; p = 0.024). CONCLUSION: Medical and surgical Menière's patients were similar utilizing academy classification. There was no significant trend between medical and surgical Menière's patients versus controls for any measurement. In a combined Menière's group, the longer and narrower vestibule anatomy may suggest an anatomical basis for endolymphatic hydrops.

Inhibition of Hedgehog-Signaling Driven Genes in Prostate Cancer Cells by Sutherlandia frutescens Extract.

Sutherlandia frutescens (L) R. Br. (Sutherlandia) is a South African botanical that is traditionally used to treat a variety of health conditions, infections and diseases, including cancer. We hypothesized Sutherlandia might act through Gli/ Hedgehog (Hh)-signaling in prostate cancer cells and used RNA-Seq transcription profiling to profile gene expression in TRAMPC2 murine prostate cancer cells with or without Sutherlandia extracts. We found 50% of Hh-responsive genes can be repressed by Sutherlandia ethanol extract, including the canonical Hh-responsive genes Gli1 and Ptch1 as well as newly distinguished Hh-responsive genes Hsd11b1 and Penk.

Inhibition of hedgehog/Gli signaling by botanicals: a review of compounds with potential hedgehog pathway inhibitory activities.

The hedgehog (Hh) signaling pathway is an important therapeutic target in cancer; involvement of the Hh pathway has been shown in a variety of cancers including basal cell carcinoma, medulloblastoma, leukemia, and gastrointestinal, breast, prostate, lung, and pancreatic cancers [1-10]. Currently, several Hh pathway inhibitory drugs are in clinical development, and the FDA recently approved Erivedge (vismodegib) from Curis/Genentech [11-15]. These new drugs are effective in many, but not all patients [16]. In fact there are documented reports of tumors developing mutations that confer resistance to the drugs [14, 17-19]. This highlights the importance of finding second generation drugs that can be used on cancers that develop resistance to the first generation Hh inhibitors. Botanicals may serve as the backbone for such research. The gold-standard pathway inhibitor, cyclopamine, is itself a naturally occurring alkaloid found in Veratrum californicum [20]. In this review we will summarize the available literature on botanical compounds in Hh-related studies. In particular we will look at curcumin, genistein, EGCG, resveratrol, quercetin, baicalen, and apigenin along with novel compounds isolated from Southeast Asian plants, such as the potent sub-micromolar gitoxigenin derivatives. Due to the nature of the pathway, most of the research published has focused on functional Gli-transcriptional assays, which we will describe and summarize.