We have previously demonstrated the
in vivo
chemopreventive efficacy of flavokawain A (FKA), a novel chalcone from the kava plant, in prostate carcinogenesis models. However, the mechanisms of the ...anticarcinogenic effects of FKA remain largely unknown. We evaluated the effect of FKA on prostate tumor spheroid formation by prostate cancer stem cells, which were sorted out from CD44+/CD133+ prostate cancer cells 22Rv1 and DU145. FKA treatment significantly decreased both the size and numbers of the tumor spheroids over different generations of spheroid passages. In addition, the dietary feeding of FKA-formulated food to Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice bearing CD44+/CD133+ 22Rv1 xenograft tumors resulted in a significant reduction of tumor growth compared to those fed with vehicle control food–fed mice. Furthermore, the expression of stem cell markers, such as Nanog, Oct4, and CD44, were markedly downregulated in both tumor spheroids and tumor tissues. We also observed that FKA inhibits Ubc12 neddylation, c-Myc, and keratin-8 expression in both CD44+/CD133+ prostate tumor spheroids and xenograft tumors. Our results suggest that FKA can reduce the tumor-initiating properties and stemness of prostate cancer, which provides a new mechanism for the chemoprevention efficacy of FKA.
Flavokawain B (FKB) has been identified from kava root extracts as a potent apoptosis inducer for inhibiting the growth of various cancer cell lines, including prostate cancer. However, the molecular ...targets of FKB in prostate cancer cells remain unknown.
An in vitro NEDD8 Initiation Conjugation Assay was used to evaluate the neddylation inhibitory activity of FKB. Molecular docking and a cellular thermal shift assay were performed to assess the direct interaction between FKB and the NEDD8 activating enzyme (NAE) complex. Protein neddylation, ubiqutination, stability and expression in cells were assessed with immunoprecipitation and Western blotting methods using specific antibodies. Deletion and site specific mutants and siRNAs were used to evaluate deep mechanisms by which FKB induces Skp2 degradation. Cell growth inhibition and apoptosis induction were measured by MTT, ELISA and Western blotting methods.
FKB inhibits NEDD8 conjugations to both Cullin1 and Ubc12 in prostate cancer cell lines and Ubc12 neddylation in an in vitro assay. Molecular docking study and a cellular thermal shift assay reveal that FKB interacts with the regulatory subunit (i.e. APP-BP1) of the NAE. In addition, FKB causes Skp2 degradation in an ubiquitin and proteasome dependent manner. Overexpression of dominant-negative cullin1 (1-452), K720R mutant (the neddylation site) Cullin1 or the F-box deleted Skp2 that losses its binding to the Skp1/Cullin1 complex causes the resistance to FKB-induced Skp2 degradation, whereas siRNA knock-down of Cdh1, a known E3 ligase of Skp2 for targeted degradation, didn't attenuate the effect of FKB on Skp2 degradation. These results suggest that degradation of Skp2 by FKB is involved in a functional Cullin1. Furthermore, proteasome inhibitors Bortezomib and MG132 transcriptionally down-regulate the expression of Skp2, and their combinations with FKB result in enhanced inhibitory effects on the growth of prostate cancer cell lines via synergistic down-regulation of Skp2 and up-regulation of p27/Kip1 and p21/WAF1 protein expression. FKB also selectively inhibits the growth of RB deficient cells with high expression of Skp2.
These findings provide a rationale for further investigating combination of FKB and Bortezomib for treatment of RB deficient, castration-resistant prostate cancer.
Efficient large-scale and feasible industrial synthesis of the 1-oxacephem core structure from 6-aminopenicillanic acid (6-APA) has been reported for several decades. Via the industrial synthesis ...route, a byproduct (compound 9) containing a butenolide unit was purified and characterized by NMR and HRMS in this work. It is worth noting that compound 9 is an entirely new compound. Additionally, a plausible mechanism and effects on the formation of 9 by different Lewis acids were proposed. The discovery of compound 9 could improve the purity of this feasible industrial synthesis and provide considerable cost savings.
Gartanin, a 4‐prenylated xanthone, has been identified from the purple mangosteen fruit as a potent growth inhibitor of various cancer cell lines, including prostate cancer. However, much of ...Gartanin's anticancer mechanism remains unknown. We have discovered that Gartanin docked onto the regulatory subunit of the precursor cell‐expressed developmentally downregulated 8 (NEDD8)‐activating enzyme (NAE) complex and next to the NEDD8 binding complex, which leads to inhibit NEDD8 conjugations to both Cullin1 and Ubc12 in prostate cancer cell lines and Ubc12 NEDDylation in an in vitro assay. The S phase kinase‐associated protein (Skp2) and F‐box and WD‐repeat domain‐containing 2 (FBXW2), the NEDD8 family members of E3 ubiqutin ligases, were also downregulated and upregulated by Gartainin, respectively. Knock‐down of NEDD8 expression by short harpin (Sh) RNAs blocked or attenuated these effects of Gartainin. Finally, Gartanin demonstrated its ability to inhibit growth of prostate cancer lines via autophagy initiation. Our data support that Gartanin is a naturally occurring NEDDylation inhibitor and deserves further investigation for prostate cancer prevention and treatment.
Novel bioactive heterocycles containing a 3,4,5-trimethoxyphenyl fragment as antiproliferative agents by targeting tubulin were synthesized and their preliminary structure activity relationships ...(SARs) were explored. Among all these chemical agents, 2-(Benzodoxazol-2-ylthio)-N-(4-methoxybenzyl)-N-(3,4,5-trimethoxyphenyl)acetamide (4d) exhibited the potent antiproliferative activity against MGC-803 cells with an IC50 value of 0.45 μM by induction of G2/M pahse arrest and cell apoptosis. In addition, 4d could change the membrane potential (ΔΨ) of the mitochondria against MGC-803 cells. Importantly, 4d acted as a novel tubulin polymerization inhibitor binding to colchicine site with an IC50 value of 3.35 μM.
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•4d exhibited the potent antiproliferative activity against MGC-803 cells.•4d induced cell cycle G2/M phase arrest and apoptosis.•4d changed the mitochondrial membrane potential against MGC-803 cells.•4d acted as a novel colchicine site tubulin polymerization inhibitor.
Here, we aim to evaluate the chemopreventive efficacy of kava root extracts (KRE) in transgenic adenocarcinoma of the mouse prostate (TRAMP) mice and investigate potential molecular targets of ...kavalactones, the main components of kava.
TRAMP mice were administrated with KRE formulated food for different periods of time, and then the incidences of high-grade prostatic intraepithelial neoplasia (HG-PIN) and adenocarcinomas and tumor burdens were compared between vehicle control and KRE food fed groups. In addition, the inhibitory effect of the KRE and kavalactones on monoamine oxidase A (MAO-A) and lysine-specific demethylase 1 (LSD1) enzyme activities were examined by commercially available inhibitor screening kits. Histone H3 lysine 9 dimethylation was also evaluated in prostate cancer cells and tumor tissues using Western blotting analysis.
Dietary feeding of 0.3% and 0.6% KRE to TRAMP mice from ages of 6 weeks to 12 weeks inhibited HG-PIN by 43.5% and 59.7%, respectively, and prostate adenocarcinoma by 53.5% and 66.4%, respectively. In addition, 0.6% KRE fed TRAMP mice from ages of 6 weeks to 24 weeks exhibited a significant reduction of genitourinary weight (a surrogate of tumor burden) by 54.5% and reduced body weight gain. Furthermore, the KRE and kavalactones showed a significant inhibition of LSD1 and MAO-A enzyme activities.
Our results suggest that consumption of kava products through diet can delay prostate cancer development and progression and that kavalactones may be a new structure model for developing a potent dual inhibitor of LSD1 and MAO-A.
This study examined if individuals who are at increased risk for drowsy driving because of obstructive sleep apnea syndrome (OSAS), have impairments in driving performance in the moments during ...microsleep episodes as opposed to during periods of wakefulness. Twenty-four licensed drivers diagnosed with OSAS based on standard clinical and polysomnographic criteria, participated in an hour-long drive in a high-fidelity driving simulator with synchronous electroencephalographic (EEG) recordings for identification of microsleeps. The drivers showed significant deterioration in vehicle control during the microsleep episodes compared to driving performance in the absence of microsleeps on equivalent segments of roadway. The degree of performance decrement correlated with microsleep duration, particularly on curved roads. Results indicate that driving performance deteriorates during microsleep episodes. Detecting microsleeps in real-time and identifying how these episodes of transition between wakefulness and sleep impair driver performance is relevant to the design and implementation of countermeasures such as drowsy driver detection and alerting systems that use EEG technology.
•Naturalistic driving data differentiates drivers with and without sleep apnea.•Topic models identified patterns, called topics, in the speed and acceleration data.•Topics distinguished drivers based ...on lateral acceleration at mid to low speeds.•Random forest models using the topics identified trips from drivers with sleep apnea.•Topic models provide a more holistic description than measures of central tendency.
One challenge in using naturalistic driving data is producing a holistic analysis of these highly variable datasets. Typical analyses focus on isolated events, such as large g-force accelerations indicating a possible near-crash. Examining isolated events is ill-suited for identifying patterns in continuous activities such as maintaining vehicle control. We present an alternative approach that converts driving data into a text representation and uses topic modeling to identify patterns across the dataset. This approach enables the discovery of non-linear patterns, reduces the dimensionality of the data, and captures subtle variations in driver behavior. In this study topic models were used to concisely described patterns in trips from drivers with and without untreated obstructive sleep apnea (OSA). The analysis included 5000 trips (50 trips from 100 drivers; 66 drivers with OSA; 34 comparison drivers). Trips were treated as documents, and speed and acceleration data from the trips were converted to “driving words.” The identified patterns, called topics, were determined based on regularities in the co-occurrence of the driving words within the trips. This representation was used in random forest models to predict the driver condition (i.e., OSA or comparison) for each trip. Models with 10, 15 and 20 topics had better accuracy in predicting the driver condition, with a maximum AUC of 0.73 for a model with 20 topics. Trips from drivers with OSA were more likely to be defined by topics for smaller lateral accelerations at low speeds. The results demonstrate topic modeling as a useful tool for extracting meaningful information from naturalistic driving datasets.
•Some actigraphic measures of sleep quality remain disturbed in patients with OSA after 3 months of therapy in PAP-adherent individuals.•Overt sleepiness as indexed by ESS normalizes with PAP, but ...FOSQ shows evidence of sleepiness-related quality-of-life impairments in patients.•Subjective symptoms as reported in ESS and FOSQ improve slowly after introduction of PAP.
Some patients with obstructive sleep apnea (OSA) remain sleepy despite positive airway pressure (PAP) therapy. The mechanisms by which this occurs are unclear but could include persistently disturbed sleep. The goal of this study was to explore the relationships between subjective sleepiness and actigraphic measures of sleep during the first three months of PAP treatment.
We enrolled 80 patients with OSA and 50 comparison subjects prior to treatment and observed them through three months of PAP therapy. PAP adherence and presence of residual respiratory events were determined from PAP machine downloads. Epworth Sleepiness Scale (ESS), Functional Outcomes of Sleep Questionnaire (FOSQ), and actigraphic data were collected before and at monthly intervals after starting PAP.
Patients with OSA were sleepier and showed a greater degree of sleep disruption by actigraphy at the baseline. After three months of PAP, only ESS and number of awakenings (AWAKE#) normalized, while wake after sleep onset and sleep efficiency remained worse in patients with OSA. FOSQ was improved in patients with OSA but never reached the same level as that of comparison subjects. ESS and FOSQ improved slowly over the study period.
As a group, patients with OSA show actigraphic evidence of persistently disturbed sleep and sleepiness-related impairments in day-to-day function after three months of PAP therapy. Improvements in sleepiness evolve over months with more severely affected patients responding quicker. Persistent sleep disruption may partially explain residual sleepiness in some PAP-adherent OSA patients.
Abstract Objective To determine the effects of obstructive sleep apnea (OSA) on visual vigilance during simulated automobile driving. Methods Twenty-five drivers with OSA and 41 comparison drivers ...participated in an hour-long drive in a high-fidelity driving simulator. Drivers responded to light targets flashed at seven locations across the forward horizon. Dependent measures were percent correct hit rate (HR) and reaction time (RT). Self-assessment of sleepiness used the Stanford Sleepiness Scale (SSS) before and after the drive and the Epworth Sleepiness Scale (ESS). Results OSA drivers showed reduced vigilance based on lower HR than comparison drivers, especially for peripheral targets (80.7±14.8% vs. 86.7±8.8%, P =.03). OSA drivers were sleepier at the end of the drive than comparison drivers (SSS=4.2±1.2 vs. 3.6±1.2, P =.03), and increased sleepiness correlated with decreased HR only in those with OSA ( r =−0.49, P =.01). Lower HR and higher post-drive SSS predicted greater numbers of driving errors in all subjects. Yet, ESS, predrive SSS, and most objective measures of disease severity failed to predict driving and vigilance performance in OSA. Conclusions Reduced vigilance for peripheral visual targets indicates that OSA drivers have restriction of their effective field of view, which may partly explain their increased crash risk. This fatigue-related decline in attention is predicted by increased subjective sleepiness during driving. These findings may suggest a means of identifying and counseling high-risk drivers and aid in the development of in-vehicle alerting and warning devices.
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