Black raspberry restores the expression of the tumor suppressor p120ctn in the oral cavity of mice treated with the carcinogen dibenzo[a,l]pyrene diol epoxide

One of the major risk factors for head and neck squamous cell carcinoma (HNSCC) is tobacco smoke exposure, but the mechanisms that can account for disease development remain to be fully defined. Utilizing our HNSCC mouse model, we analyzed oral squamous cell carcinomas (OSCC) induced by the active metabolite of a common smoke constituent, dibenzo[a,l]pyrene diol-epoxide (DBPDE). Analyzing protein expression by either immunofluorescence or immunohistochemistry, we identified biologic processes that are dysregulated in premalignant and invasive cancer lesions induced by DBPDE. Interestingly, p120ctn expression is downregulated in both stages of the disease. In addition to decreased p120ctn expression, there was also increased proliferation (as measured by Ki67), inflammation (as measured by NFkB (p65) expression), neovascularization (as measured by CD31) and recruitment of Ly6G-positive immune cells as well as strong EGFR expression. We also examined the effect of the chemopreventive agent black raspberry (BRB) on p120ctn and EGFR protein expression in DBPDE treated mice. p120ctn, but not EGFR, protein expression increased in mice treated with BRB. Our results suggest that modulation of p120ctn may, in part, account for the mechanism by which BRB inhibits DBPDE induced OSCC in mice.

Triacylglycerol and Fatty Acid Compositions of Blackberry, Red Raspberry, Black Raspberry, Blueberry and Cranberry Seed Oils by Ultra-Performance Convergence Chromatography-Quadrupole Time-of-Flight Mass Spectrometry

The triacylglycerol (TAG) compositions of blackberry, red raspberry, black raspberry, blueberry and cranberry seed oils were examined using ultra-performance convergence chromatography-quadrupole time-of-flight mass spectrometry (UPC2-QTOF MS). A total of 52, 53, 52, 59 and 58 TAGs were detected and tentatively identified from the blackberry, red raspberry, black raspberry, blueberry and cranberry seed oils, respectively, according to their accurate molecular weight in MS1 and fragment ion profiles in MS2. OLL was the most abundant TAG in the blackberry, red raspberry and black raspberry seed oils. Furthermore, the fatty acid compositions of the five berry seed oils were directly determined by gas chromatography coupled with mass spectrometry (GC-MS). In addition, the seed oils had total phenolic contents ranging 13.68–177.06 µmol GAE (gallic acid equivalent)/L oil, and significant scavenging capacities against DPPH, peroxyl, and ABTS+ radicals. These results indicated that the combination of UPC2 and QTOF MS could effectively identify and semi-quantify the TAGs compositions of the berry seed oils with sn-position information for the fatty acids. Understanding the TAGs compositions of these berry seed oils could improve the utilization of these potentially high nutritional value oils for human health.

Evaluation of Probiotic Properties of Pediococcus acidilactici M76 Producing Functional Exopolysaccharides and Its Lactic Acid Fermentation of Black Raspberry Extract

This study aimed to determine the probiotic potential of Pediococcus acidilactici M76 (PA-M76) for lactic acid fermentation of black raspberry extract (BRE). PA-M76 showed outstanding probiotic properties with high tolerance in acidic GIT environments, broad antimicrobial activity, and high adhesion capability in the intestinal tract of Caenorhabditis elegans. PA-M76 treatment resulted in significant increases of pro-inflammatory cytokine mRNA expression in macrophages, indicating that PA-M76 elicits an effective immune response. When PA-M76 was used for lactic acid fermentation of BRE, an EPS yield of 1.62 g/L was obtained under optimal conditions. Lactic acid fermentation of BRE by PA-M76 did not significantly affect the total anthocyanin and flavonoid content, except for a significant increase in total polyphenol content compared to non-fermented BRE (NfBRE). However, fBRE exhibited increased DPPH radical scavenging activity, linoleic acid peroxidation inhibition rate, and ABTS scavenging activity of fBRE compared to NfBRE. Among the 28 compounds identified in the GC-MS analysis, esters were present as the major groups. The total concentration of volatile compounds was higher in fBRE than that in NfBRE. However, the undesirable flavor of terpenes decreased. PA-M76 might be useful for preparing functionally enhanced fermented beverages with a higher antioxidant activity of EPS and enhanced flavors.

Kuehneola uredinis (cane and leaf rust).

Kuehneola uredinis is a fungal pathogen that causes cane and leaf rust only in Rubus cultivars, or wild and ornamental blackberry species. It can be highly destructive, especially on susceptible blackberry and Rubus hybrid cultivars, but rarely occurs on red and black raspberry (Ellis et al., 1991). Although the disease is not systemic, severe infection can cause premature defoliation and a decrease in cane vigour, making plants more susceptible to winter conditions and resulting in economic loss (Converse, 1966). Dispersal of the pathogen occurs on blackberry and hybrid Rubus cultivars, and wild blackberries in production areas worldwide. The fungus overwinters either as aecial urediniospores or mycelium in the canes and wet conditions favour disease dispersal (Laundon and Rainbow, 1969). K. uredinis is not included in quarantine lists (USDA, 2017; EPPO, 2019). There is interest in K. uredinis as a biocontrol agent due to its destructive effect on some invasive wild blackberry species, e.g., Rubus penetrans (Gardner and Hodges, 1983).

Protective Effects of Berry Volatiles on Lipopolysaccharide-Induced Acute Lung Injury in Human A549 Cells

Objectives Berry volatiles are primary or secondary compounds produced when tissue disrupts, and responsible for the flavor and aroma of berries. Berry volatiles has been recently reported to have anti-inflammatory effect. Acute lung injury is one of the severe consequences of lung inflammation increasing respiratory stress and pulmonary edema with high mortality. Thus, the objective of this study was to investigate the anti-inflammatory effect of three berry volatiles (blackberry, black raspberry, and blueberry) on lipopolysaccharide (LPS)-induced acute lung injury in lung epithelial A549 cells and its mechanism. Methods A549 cells were pretreated with three different dilutions (2, 4, and 8-fold) of blackberry, black raspberry, and blueberry volatile extract for 1 h. Cells were then stimulated with or without LPS (10 μg/mL) for 24 h. The cell viability was measured with one-step diphenyl tetrazolium bromide (MTS) assay. The level of pro-inflammatory cytokines in the culture media was examined by using enzyme-linked immunosorbent assay (ELISA) kit. Apoptosis of A549 cells were evaluated by using death detection ELISA kit. All statistical tests were analyzed by using one-way ANOVA, followed by Tukey's multiple comparisons. Significant difference was defined at P < 0.05. Results Two-fold diluted berry volatiles (blackberry, black raspberry, and blueberry) significantly decreased A549 cell viability by 24%, 52% and 61%, respectively compared to the control (P < 0.05). Concentration of interleukin-8 (IL-8) was remarkably higher when LPS stimulated A549 cells compared to the control (P < 0.05). However, 2-fold diluted berry volatiles significantly suppressed LPS-induced IL-8 level compared to LPS treated group (P < 0.01). 2-fold diluted blackberry, black raspberry, and blueberry volatiles also showed significant apoptotic effect on A549 cells compared to the control (P < 0.05). Conclusions These results showed that blackberry, black raspberry, and blueberry volatiles suppress inflammatory responses by modulating pro-inflammatory cytokines and apoptosis. This study suggests that volatiles from berries (blackberry, black raspberry, and blueberry) may have a potential impact on lung inflammation. Funding Sources Arkansas Biosciences Institute.

Kuehneola uredinis (cane and leaf rust).

Kuehneola uredinis is a fungal pathogen that causes cane and leaf rust only in Rubus cultivars, or wild and ornamental blackberry species. It can be highly destructive, especially on susceptible blackberry and Rubus hybrid cultivars, but rarely occurs on red and black raspberry (Ellis et al., 1991). Although the disease is not systemic, severe infection can cause premature defoliation and a decrease in cane vigour, making plants more susceptible to winter conditions and resulting in economic loss (Converse, 1966). Dispersal of the pathogen occurs on blackberry and hybrid Rubus cultivars, and wild blackberries in production areas worldwide. The fungus overwinters either as aecial urediniospores or mycelium in the canes and wet conditions favour disease dispersal (Laundon and Rainbow, 1969). K. uredinis is not included in quarantine lists (USDA, 2017; EPPO, 2019). There is interest in K. uredinis as a biocontrol agent due to its destructive effect on some invasive wild blackberry species, e.g., Rubus penetrans (Gardner and Hodges, 1983).

Black raspberry anthocyanins protect BV2 microglia from LPS-induced inflammation through down-regulating NOX2/TXNIP/NLRP3 signaling

BACKGROUND: Increasing evidence has established neuroinflammation as the hallmark of neurodegenerative disorders such as Alzheimer’s disease (AD). However, despite the underlying immunological mechanisms are far from being understood, the involvement of excessive activation of microglia is attracting more and more attention. OBJECTIVE: In the present study, we investigated the protective effect of black raspberry (BRB) anthocyanins on LPS-induced neuroinflammation in BV2 microglia. METHODS: LPS-induced mouse BV2 microglia were treated with black raspberry anthocyanins and the levels of NO, ROS, IL-1β and IL-18 produced by the cells were measured to determine the extent of oxidative stress and inflammatory response. RESULTS: The results showed that BRB anthocyanins reduced the production of ROS in LPS-induced BV2 microglia by down-regulating the level of NOX2 and its downstream factors, including thioredoxin-interacting protein (TXNIP) and NOD-like receptor protein 3 (NLRP3) inflammasome. Furthermore, BRB anthocyanins inhibited the secretion of Interleukin-18 (IL-18) and Interleukin-1β (IL-1β), eventually attenuating the LPS-induced inflammatory response of BV2 microglia. CONCLUSIONS: BRB anthocyanins might play an important neuroprotective role in inflammation-related neurodegenerative disease, potentially, by down-regulating the NOX2 /TXNIP/ NLRP3 signaling axis in brain microglia.