Jennifer Ahn-Jarvis

Characterization of black raspberry functional food products for cancer prevention human clinical trials.

Our team is designing and fully characterizing black raspberry (BRB) food products suitable for long-term cancer prevention studies. The processing, scale-up, and storage effects on the consistency, quality, bioactive stability, and sensory acceptability of two BRB delivery systems of various matrices are presented. BRB dosage, pH, water activity, and texture were consistent in the scale-up production. Confections retained >90% of anthocyanins and ellagitannin after processing. Nectars had >69% of anthocyanins and >66% of ellagitannin retention, which varied with BRB dosage due to the processing difference. Texture remained unchanged during storage. BRB products consumed in a prostate cancer clinical trial were well accepted in sensory tests. Thus, this study demonstrates that two different BRB foods can be formulated to meet quality standards with a consistent bioactive pattern and successfully scaled up for a large human clinical trial focusing on cancer risk and other health outcomes.

Consumption of Soy Isoflavone Enriched Bread in Men with Prostate Cancer Is Associated with Reduced Proinflammatory Cytokines and Immunosuppressive Cells

We hypothesized that soy phytochemicals may have immunomodulatory properties that may affect prostate carcinogenesis and progression. A randomized, phase II trial was conducted in 32 patients with prostate cancer with asymptomatic biochemical recurrence but no measurable disease on standard staging studies. Patients were randomized to two slices of soy bread (34 mg isoflavones/slice) or soy bread containing almond powder daily as a source of β-glucosidase. Flow cytometry and bioplex assays were used to measure cytokines or immune cell phenotype in blood at baseline (day 0) and following intervention (day 56). Adequate blood samples were available at enrollment and day 56 and evaluated. Multiple plasma cytokines and chemokines were significantly decreased on day 56 versus baseline. Subgroup analysis indicated reduced TH1 (P = 0.028) and myeloid-derived suppressor cell (MDSC)-associated cytokines (P = 0.035). TH2 and TH17 cytokines were not significantly altered. Phenotypic analysis revealed no change in CD8+ or CD4+ T cells but showed increased CD56+ natural killer (NK) cells (P = 0.038). The percentage of cells with a T regulatory cell phenotype (CD4+CD25+FoxP3+) was significantly decreased after 56 days of soy bread (P = 0.0136). Significantly decreased monocytic (CD33+HLADRnegCD14+) MDSC were observed in patients consuming soy bread (P = 0.0056). These data suggest that soy bread modulates systemic soluble and cellular biomarkers consistent with limiting inflammation and suppression of MDSCs. Additional studies to elucidate impact on the carcinogenic process or as a complement to immune-based therapy are required. Cancer Prev Res; 8(11); 1036–44. ©2015 AACR.

Design and Selection of Soy Breads Used for Evaluating Isoflavone Bioavailability in Clinical Trials

To modulate isoflavone aglycone composition within a soy functional food, soy ingredients were processed and evaluated in a soy bread system intended for clinical trials. A soy flour/soy milk mixture (SM) was boiled, fermented, steamed, or roasted prior to dough preparation. The isoflavone compositions of five processed SM and their corresponding breads combined with and without β-glucosidase-rich almonds were examined using HPLC. Isoflavone malonyl-glucosides (>80%) were converted into acetyl and simple glucoside forms (substrates more favorable for β-glucosidase) in steamed and roasted SM. Their corresponding breads had isoflavones predominately as aglycones (∼75%) with soy-almond bread with steamed SM being more consumer acceptable than roasted. Isoflavone composition in soy bread was stable during frozen storage and toasting. A suitable glycoside-rich soy bread (31.6 ± 2.1 mg aglycone equiv/slice) using unprocessed SM and an aglycone-rich soy-almond bread (31.1 ± 1.9 mg aglycone equiv/slice) using steamed SM were developed to evaluate fundamental questions of isoflavone bioavailability in clinical trials.

Isoflavone pharmacokinetics and metabolism after consumption of a standardized soy and soy-almond bread in men with asymptomatic prostate cancer

Epidemiologic associations suggest that populations consuming substantial amounts of dietary soy exhibit a lower risk of prostate cancer. A 20-week randomized, phase II, crossover trial was conducted in 32 men with asymptomatic prostate cancer. The crossover involved 8 weeks each of soy bread (SB) and soy–almond bread (SAB). The primary objective was to investigate isoflavone bioavailability and metabolite profile. Secondary objectives include safety, compliance, and assessment of biomarkers linked to prostate carcinogenesis. Two distinct SBs were formulated to deliver approximately 60 mg aglycone equivalents of isoflavones per day. The isoflavones were present as aglycones (∼78% as aglycones) in the SAB whereas in the standard SB predominantly as glucosides (18% total isoflavones as aglycones). Compliance to SB (97% ± 4%) and SAB (92% ± 18%) was excellent; toxicity was rare and limited to grade 1 gastrointestinal complaints. Pharmacokinetic studies between SB and SAB showed modest differences. Peak serum concentration time (Tmax) was significantly faster with SAB meal compared with SB in some isoflavonoids, and AUC0 to 24 h of dihydrodaidzein and O-desmethylangolensin was significantly greater after an SB meal. An exploratory cluster analysis was used to identify four isoflavone-metabolizing phenotypes. Insulin-like growth factor–binding protein increased significantly by 41% (P = 0.024) with soy intervention. Findings from this study provide the necessary framework to study isoflavone-metabolizing phenotypes as a strategy for identification of individuals that might benefit or show resistance to cancer preventive strategies using dietary soy. A standardized SB used for future large-scale randomized clinical trials to affect human prostate carcinogenesis is feasible. Cancer Prev Res; 8(11); 1045–54. ©2015 AACR.

Inhibition of Pro-inflammatory and Anti-apoptotic Biomarkers during Experimental Oral Cancer Chemoprevention by Dietary Black Raspberries

Oral cancer continues to be a significant public health problem worldwide. Recently conducted clinical trials demonstrate the ability of black raspberries (BRBs) to modulate biomarkers of molecular efficacy that supports a chemopreventive strategy against oral cancer. However, it is essential that a preclinical animal model of black raspberry (BRB) chemoprevention which recapitulates human oral carcinogenesis be developed, so that we can validate biomarkers and evaluate potential mechanisms of action. We therefore established the ability of BRBs to inhibit oral lesion formation in a carcinogen-induced rat oral cancer model and examined potential mechanisms. F344 rats were administered 4-nitroquinoline 1-oxide (4NQO) (20 µg/ml) in drinking water for 14 weeks followed by regular drinking water for 6 weeks. At week 14, rats were fed a diet containing either 5 or 10% BRB, or 0.4% ellagic acid (EA), a BRB phytochemical. Dietary administration of 5 and 10% BRB reduced oral lesion incidence and multiplicity by 39.3 and 28.6%, respectively. Histopathological analyses demonstrate the ability of BRBs and, to a lesser extent EA, to inhibit the progression of oral cancer. Oral lesion inhibition by BRBs was associated with a reduction in the mRNA expression of pro-inflammatory biomarkers Cxcl1, Mif, and Nfe2l2 as well as the anti-apoptotic and cell cycle associated markers Birc5, Aurka, Ccna1, and Ccna2. Cellular proliferation (Ki-67 staining) in tongue lesions was inhibited by BRBs and EA. Our study demonstrates that, in the rat 4NQO oral cancer model, dietary administration of BRBs inhibits oral carcinogenesis via inhibition of pro-inflammatory and anti-apoptotic pathways.

Physicochemical Characterization and Sensory Analysis of Yeast-leavened and Sourdough Soy Breads

Sourdough fermentation has been shown to have numerous beneficial effects on bread quality, and nutritionally enhance soy-supplemented bread by altering isoflavone chemical forms. Given this, the objective of this study was to compare the loaf quality and shelf life of sourdough and yeast-leavened soy breads by various physical, thermal, and sensorial methods, and to assess the effects of fermentation by various microorganisms on isoflavone profile in dough and breads using high-performance liquid chromatography analysis. Sourdough fermentation yielded a less extensible dough compared to yeast-leavened soy dough (P < 0.001), and resulted in a harder bread crumb (P < 0.05) and lighter crust color (P < 0.001), compared to yeast-leavened soy bread (Y-B). Sensory analysis revealed a significantly higher overall liking of Y-B compared to sourdough soy bread (SD-B) (P < 0.001). Segmentation analysis of the cohort suggests that overall liking and bread consumption frequency may be determinants of Y-B or SD-B preference. SD-B and Y-B exhibited similar shelf-life properties. Despite significantly different enthalpies associated with the melting of amylose-lipid complexes, thermal analysis of the 2 soy breads stored for 10 d (ambient conditions) demonstrated no significant difference in water distribution and starch retrogradation (P < 0.05). Lastly, SD-B was determined to have 32% of total isoflavones occurring in the aglycone form compared to 17% in Y-B. These findings warrant further investigation of sourdough fermentation as a processing technique for quality and nutritional enhancement of soy-based baked goods.

Pediatric Cutaneous Leishmaniasis in an Endemic Region in Turkey: A Retrospective Analysis of 8786 Cases during 1998-2014.

Background Cutaneous leishmaniasis (CL) is a major public health concern in Turkey and Sanliurfa represents the most endemic city in Turkey. Although children are most commonly affected by CL, detailed studies of pediatric CL in Turkey are lacking. Methodology/Principal Findings In this report we retrospectively evaluated clinical and epidemiological data of 8786 pediatric CL cases, and how children respond to antimonial therapy. CL was observed most frequently in children between 6–10 years old. Interestingly this group showed shorter duration of disease and smaller lesions compared to 0–5 year and 11–15 year old groups. Females were more affected in all groups. Lesion localization and types varied among groups, with 0–5 year old presenting head/neck and mucosal lesions, and more often suffered from recidivans type, this could be associated to the longest duration of the disease in this group. Eleven-15 year old group showed fewer lesions in the head/neck but more generalized lesions. Evaluation of treatment response revealed that intra-lesional treatment was preferred over intramuscular treatment. However, 0–5 year old received intramuscular treatment more often than the other groups. Furthermore, the majority of 0–5 year old group which received intra-lesional treatment did not received subsequent intra-lesional cycles, as did children in the range of 6–15 years old. Conclusions/Significance We report an increase in pediatric CL patients within the last four years. Analysis of pediatric CL patients by age revealed significant differences in CL progression. The data suggest that children between 0–5 years old responded better than other groups to intralesional treatment, since they received more often a single cycle of IL treatment, although follow up observation is required since they were more prone to develop recidivans. Eleven-15 year old patients comprise the largest percentage of patients receiving two or three cycles of intralesional treatment, suggesting that this group did not respond efficiently to intralesional treatment and highlighting the need for more effective therapeutic strategies against CL.

Deletion of macrophage migration inhibitory factor inhibits murine oral carcinogenesis: Potential role for chronic pro-inflammatory immune mediators.

Oral cancer kills about 1 person every hour each day in the United States and is the sixth most prevalent cancer worldwide. The pro‐inflammatory cytokine ‘macrophage migration inhibitory factor’ (MIF) has been shown to be expressed in oral cancer patients, yet its precise role in oral carcinogenesis is not clear. In this study, we examined the impact of global Mif deletion on the cellular and molecular process occurring during oral carcinogenesis using a well‐established mouse model of oral cancer with the carcinogen 4‐nitroquinoline‐1‐oxide (4NQO). C57BL/6 Wild‐type (WT) and Mif knock‐out mice were administered with 4NQO in drinking water for 16 weeks, then regular drinking water for 8 weeks. Mif knock‐out mice displayed fewer oral tumor incidence and multiplicity, accompanied by a significant reduction in the expression of pro‐inflammatory cytokines Il‐1β, Tnf‐α, chemokines Cxcl1, Cxcl6 and Ccl3 and other molecular biomarkers of oral carcinogenesis Mmp1 and Ptgs2. Further, systemic accumulation of myeloid‐derived tumor promoting immune cells was inhibited in Mif knock‐out mice. Our results demonstrate that genetic Mif deletion reduces the incidence and severity of oral carcinogenesis, by inhibiting the expression of chronic pro‐inflammatory immune mediators. Thus, targeting MIF is a promising strategy for the prevention or therapy of oral cancer.

Modulating conversion of isoflavone glycosides to aglycones using crude beta-glycosidase extracts from almonds and processed soy

Food processing alters the physicochemical state of soy which can enhance chemical and enzymatic conversion of isoflavones to their aglycone forms. This study investigated the role of β-glycosidase from processed soy-ingredient mixture (SIM) or almonds, and examined the impact of isoflavone composition in mediating conversion to aglycones. β-Glycosidase activity was quantified using p-nitrophenol-β-d-glucopyranoside and SIM isoflavone extracts. Almond β-glycosidase activity was significantly (p<0.001) reduced after roasting (99% reduction) or steaming (97% reduction) compared to raw almonds. SIM β-glycosidase activity, however, increased, with steaming by 66% (p<0.001) and with roasting by 52% (p=0.022), compared to raw SIM. After incubation with β-glycosidase, percentage of aglycone (total aglycone/total isoflavones) in SIM isoflavone extracts increased significantly in raw (35%), fermented (48%), roasted (88%) and steamed (91%) SIM, compared to their initial (∼5%) compositions. Manipulation of β-glycosidase activity and isoflavone composition can be used to modulate aglycone content in soy food products.

Abstract CT105: Validation of a tobacco smoke exposure gene expression signature and exploration of intraoral metabolite profiles following administration of a strawberry functional confection in smokers and nonsmokers

Dietary administration of whole strawberries has demonstrated great potential as a strategy for oral and esophageal cancer prevention in preclinical trials. We hypothesize that following consumption of a novel confection containing strawberries (i) gene expression profiles in oral mucosa will be modulated in a manner that favor anti-cancer activities, and (ii) unique intraoral metabolites will segregate smokers and nonsmokers. A 6-week randomized, placebo-controlled, Phase I crossover trial using functional confections delivering 24g/day of freeze-dried whole ripe strawberries was examined in smoking and nonsmoking men and women. Objectives Validate an annotated targeted oral epithelial gene expression profile in smokers and nonsmokers. Establish a strawberry-associated oral epithelial gene expression profile in smokers and nonsmokers following localized delivery of a novel strawberry functional confection. Define an intraoral metabolic profile of strawberry compounds in saliva for smokers and nonsmokers. Methods: Total RNA was isolated from tongue oral epithelium brush biopsies and gene expression was measured using RT-qPCR analyses with 44 pre-validated TaqMan gene expression assays corresponding to known tobacco-smoke associated oral transcriptome biomarkers. Anthocyanin and ellagitannins as well as other strawberry metabolites in saliva were quantified using HPLC with photodiode-array and UPLC with tandem mass spectroscopy. Salivary amylase and beta-glucosidase activity was measured colorimetrically using p-nitrophenol endpoints. Results: A 7-gene over-expression signature (ALOX12B, CD207, HTR3A, KRT10, LOR, PNLIPRP3, TRNP1) was validated that segregated smokers and nonsmokers (adjusted p-value Conclusions: Significant differences between smokers and nonsmokers were observed in gene expression, salivary enzyme activity and in intraoral strawberry metabolites following administration of a strawberry functional confection. While this pilot study interrogated an existing tobacco smoke driven transcriptional profile, the global strawberry bioactive driven transcriptome remains uncharacterized. Moreover, complementary changes in oral epithelial gene expression and intraoral metabolites between smokers and nonsmokers warrant the need for long-term cancer prevention studies using strawberries. Citation Format: Jennifer H. Ahn-Jarvis, Thomas J. Knobloch, Steve Oghumu, Ken M. Reidl, Guy Brock, Steven K. Clinton, Yael Vodovotz, Steven J. Schwartz, Christopher M. Weghorst. Validation of a tobacco smoke exposure gene expression signature and exploration of intraoral metabolite profiles following administration of a strawberry functional confection in smokers and nonsmokers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT105. doi:10.1158/1538-7445.AM2017-CT105