Lycopene Accumulation in Transgenic Mice Lacking One or Both Carotenoid Cleaving Enzymes.

BACKGROUND: ß-carotene oxygenase 1 (BCO1) and ß-carotene oxygenase 2 (BCO2) are responsible for the cleavage of carotenoids in mammals. OBJECTIVE: The goals of this study were to (1) establish the relative contribution of each enzyme on lycopene accumulation in mice and (2) examine the role of lycopene on gene expression in the gut of wild type (WT) mice. METHODS: We utilized male and female WT, Bco1-/-, Bco2-/-, and Bco1-/-Bco2-/- double knockout (DKO) mice. We gavaged the mice with either 1 mg of lycopene resuspended in cottonseed oil or vehicle as a control group daily for 2 wk. In a second study, we evaluated the effect of dietary vitamin A on lycopene absorption and intestinal gene expression by RT-PCR. We also quantified lycopene concentration isomer distribution by high performance liquid chromatography. RESULTS: Of the 11 tissues measured, the liver accounted for 94 to 98% of the lycopene content across genotypes. We did not observe sex differences between genotypes, although hepatic lycopene levels in Bco1-/- mice were approximately half in comparison to the other genotypes; Bco1-/- verses Bco2-/- (P < 0.0001), DKO mice (P < 0.001), WT (ns). Analyses of mitochondrial lycopene content revealed a 3- to 5-fold enrichment compared with total hepatic content (P < 0.05) in all genotypes and sexes. In our second study, WT mice fed a vitamin A-deficient diet (VAD) accumulated greater amounts of lycopene in the liver than those fed a vitamin A-sufficient diet (VAS) (P < 0.01). These changes were accompanied by an upregulation of the vitamin A-responsive transcription factor intestine specific homeobox (ISX) in mice fed VAD + lycopene and VAS + lycopene diets compared with VAD control-fed mice (P < 0.05). CONCLUSIONS: Our data suggest that BCO2 is the primary lycopene cleavage enzyme in mice. Lycopene concentration was enriched in the mitochondria of hepatocytes independently of genotype, and lycopene stimulated vitamin A signaling in WT mice.

Understanding quality improvement collaboratives through an implementation science lens.

Quality improvement collaboratives (QICs) have long been used to facilitate group learning and implementation of evidence-based interventions (EBIs) in healthcare. However, few studies systematically describe implementation strategies linked to QIC success. To address this gap, we evaluated a QIC on colorectal cancer (CRC) screening in Federally Qualified Health Centers (FQHCs) by aligning standardized implementation strategies with collaborative activities and measuring implementation and effectiveness outcomes. In 2018, the American Cancer Society and North Carolina Community Health Center Association provided funding, in-person/virtual training, facilitation, and audit and feedback with the goal of building FQHC capacity to enact selected implementation strategies. The QIC evaluation plan included a pre-test/post-test single group design and mixed methods data collection. We assessed: 1) adoption, 2) engagement, 3) implementation of QI tools and CRC screening EBIs, and 4) changes in CRC screening rates. A post-collaborative focus group captured participants' perceptions of implementation strategies. Twenty-three percent of North Carolina FQHCs (9/40) participated in the collaborative. Health Center engagement was high although individual participation decreased over time. Teams completed all four QIC tools: aim statements, process maps, gap and root cause analysis, and Plan-Do-Study-Act cycles. FQHCs increased their uptake of evidence-based CRC screening interventions and rates increased 8.0% between 2017 and 2018. Focus group findings provided insights into participants' opinions regarding the feasibility and appropriateness of the implementation strategies and how they influenced outcomes. Results support the collaborative's positive impact on FQHC capacity to implement QI tools and EBIs to improve CRC screening rates.

Bridging Research, Practice, and Policy: The "Evidence Academy" Conference Model.

Innovative models to facilitate more rapid uptake of research findings into practice are urgently needed. Community members who engage in research can accelerate this process by acting as adoption agents. We implemented an Evidence Academy conference model bringing together researchers, health care professionals, advocates, and policy makers across North Carolina to discuss high-impact, life-saving study results. The overall goal is to develop dissemination and implementation strategies for translating evidence into practice and policy. Each 1-day, single-theme, regional meeting focuses on a leading community-identified health priority. The model capitalizes on the power of diverse local networks to encourage broad, common awareness of new research findings. Furthermore, it emphasizes critical reflection and active group discussion on how to incorporate new evidence within and across organizations, health care systems, and communities. During the concluding session, participants are asked to articulate action plans relevant to their individual interests, work setting, or area of expertise.

Development and validation of a method to deliver vitamin A to macrophages.

Macrophages are critical players in the development of atherosclerotic lesions, where they promote local and systemic inflammation. Macrophages engulf lipoproteins and cell debris upon entry into the arterial wall, becoming lipid-laden foam cells. While most lipids found in foam cells are triglyceride and cholesterol, these cells accumulate several other lipids with bioactive properties, such as vitamin A and carotenoids. Vitamin A has strong immunomodulatory actions in macrophages and other immune cells. For example, macrophages release vitamin A as retinoic acid to modulate T cell differentiation, but the implication of intracellular vitamin A stores in this process remains elusive due to the lack of an adequate experimental model to load vitamin A into macrophages. The purpose of this study was to develop a reliable method to deliver vitamin A to cultured murine macrophages. Our results show that thioglycolate-elicited peritoneal macrophages fail to take up significant levels of vitamin A when provided as free retinol. Cultured macrophages and macrophages in the peritoneal cavity can take up retinyl esters, either as retinyl ester-loaded serum or retinyl esters infused directly into the peritoneal cavity. HPLC analyses in macrophage lysates revealed that the intraperitoneal injection method results in a fourfold greater vitamin A loading efficiency than retinyl ester-loaded serum added to cultured cells. These two alternative methods provide an efficient and reliable methodology to load macrophages with vitamin A for downstream applications such as studies of gene regulation trafficking of intracellular vitamin A, and vitamin A release from macrophages.

Fenretinide inhibits vitamin A formation from ß-carotene and regulates carotenoid levels in mice.

N-[4-hydroxyphenyl]retinamide, commonly known as fenretinide, a synthetic retinoid with pleiotropic benefits for human health, is currently utilized in clinical trials for cancer, cystic fibrosis, and COVID-19. However, fenretinide reduces plasma vitamin A levels by interacting with retinol-binding protein 4 (RBP4), which often results in reversible night blindness in patients. Cell culture and in vitro studies show that fenretinide binds and inhibits the activity of ß-carotene oxygenase 1 (BCO1), the enzyme responsible for endogenous vitamin A formation. Whether fenretinide inhibits vitamin A synthesis in mammals, however, remains unknown. The goal of this study was to determine if the inhibition of BCO1 by fenretinide affects vitamin A formation in mice fed ß-carotene. Our results show that wild-type mice treated with fenretinide for ten days had a reduction in tissue vitamin A stores accompanied by a two-fold increase in ß-carotene in plasma (P < 0.01) and several tissues. These effects persisted in RBP4-deficient mice and were independent of changes in intestinal ß-carotene absorption, suggesting that fenretinide inhibits vitamin A synthesis in mice. Using Bco1-/- and Bco2-/- mice we also show that fenretinide regulates intestinal carotenoid and vitamin E uptake by activating vitamin A signaling during short-term vitamin A deficiency. This study provides a deeper understanding of the impact of fenretinide on vitamin A, carotenoid, and vitamin E homeostasis, which is crucial for the pharmacological utilization of this retinoid.

A comparison of the absorption and metabolism of the major quercetin in brassica, quercetin-3-O-sophoroside, to that of quercetin aglycone, in rats.

Although flavonoid sophorosides are common glycosides in brassica vegetables, red raspberries and other food plants, there is a lack of studies of absorption and metabolism of any sophoroside. The aim of this study was to characterize the absorption, phase II metabolism and microbial catabolism of quercetin-3-O-sophoroside, compared to that of quercetin aglycone. Quercetin-3-O-sophoroside was purified from Apocynum venetum and characterized by MS2, 1H and 13C NMR. Using an in situ rat gut model, we found intact, methylated, sulfated and both methylated and sulfated quercetin sophoroside in the plasma following jejunal introduction of the sophoroside; we found derivatives of benzoic acid, phenylacetic acid, and phenyl propionic acid in the cecal contents following cecal introduction. This novel finding, that quercetin sophoroside was absorbed intact, without deglycosylation, points to a possible role for the terminal sugar and/or the type of linkage among glycosidic moieties in the mechanism of absorption of flavonoid glycosides.

Epigenetic stratification of head and neck cancer survivors reveals differences in lycopene levels, alcohol consumption, and methylation of immune regulatory genes.

BACKGROUND: Inflammation has been associated with higher rates of recurrence and mortality in head and neck cancer (HNC). While the biological mechanisms predisposing patients to heightened inflammatory states remain largely unknown, DNA methylation has been proposed to reflect systemic inflammation. In this analysis, we attempt to identify meaningful epigenetic patterns in HNC survivors by stratifying individuals based on DNA methylation profiles in leukocytes. RESULTS: We used hierarchical clustering to uncover three distinct methylation patterns among HNC survivors. Each group displayed a unique methylation signature in inflammatory pathways including cytokine and B-cell receptor signaling. Additionally, we examined physiological, clinical, and lifestyle parameters related to inflammation, such as circulating carotenoid and cytokine levels, cancer treatment type, and alcohol consumption. Specifically, we identified one group of survivors who had significant differential methylation of transcriptional and translational regulators as well as genes in the T-cell receptor signaling pathway, including hypermethylation of CD40 ligand (CD40LG) and Tec protein tyrosine kinase (TEC) and hypomethylation of CD8A. This group also displayed high circulating lycopene levels. We identified another group that had distinctive methylation in the toll-like receptor (TLR) signaling pathway, including hypomethylation of TLR5, a component of the inhibitor of nuclear factor-kappa B kinase complex (CHUK), and two mitogen-activated protein kinases (MAP3K8 and MAP2K3). This group also had hypermethylation of mitochondrial ribosomal genes along with higher rates of alcohol consumption. CONCLUSION: The correlation between lycopene, alcohol consumption, DNA methylation, and inflammation warrants further investigation and may have implications in future recommendations and interventions to impact health outcomes in HNC survivors.

Feasibility Outcomes of a Pilot Randomized Clinical Trial to Increase Cruciferous and Green Leafy Vegetable Intake in Posttreatment Head and Neck Cancer Survivors.

BACKGROUND: Higher intakes of cruciferous vegetables (CVs) and green leafy vegetables (GLVs) in observational studies are associated with improvements in survival and cancer-related biomarkers in patients diagnosed with head and neck cancer (HNC). These results have yet to be corroborated in a randomized clinical trial (RCT). OBJECTIVE: Determine the feasibility of implementing a 12-week RCT to increase CV and GLV intake in posttreatment HNC survivors. DESIGN AND PARTICIPANTS: This was a two-arm RCT conducted among 24 posttreatment HNC survivors. Survivors were recruited from a southeastern, National Cancer Institute-designated Comprehensive Cancer Center between January 2015 and September 2016. INTERVENTION: There were two groups: (1) an experimental group (n=12) receiving weekly 15- to 30-minute telephone dietary counseling from a registered dietitian nutritionist stressing 2.5 cups per week CVs and 3.5 cups per week GLVs, and (2) an attention control group (n=12) receiving weekly 15- to 30-minute telephone dietary counseling from a registered dietitian nutritionist focusing on general healthy eating for cancer survivors. Participants completed a baseline survey, three 24-hour dietary recalls, phlebotomy, and anthropometric measures prior to randomization and at the end of the 12-week study period. The experimental group also completed weekly vegetable record recalls. MAIN OUTCOME MEASURES: Primary outcomes included feasibility, recruitment, retention, adherence, and safety. Secondary outcomes included inflammatory markers and carotenoids. STATISTICAL ANALYSES PERFORMED: Descriptive statistics were generated for demographic, epidemiological, and clinical variables as well as the primary feasibility outcomes. Between- and within-group comparisons of mean serum cytokine and carotenoid levels were performed using appropriate statistical tests depending on their respective distributions for the purpose of generating preliminary effect sizes. RESULTS: Overall, 350 incident HNC cases were screened for eligibility, and 98 were eligible for study participation. Reasons for ineligibility and exclusion included deceased (n=93); wrong or inactive telephone numbers, or unable to be reached, or lost to follow-up (n=93); not meeting inclusion criteria (n=39); and too ill to participate (n=27). Of the 98 eligible HNC cases, 24 agreed to participate, for an enrollment rate of 25%. The most common reason for nonparticipation was distance (n=48), as participants were asked to report for two on-site assignments. The retention rate was 96%. Mean intervention adherence rates for weekly goals were 67% CV, 74% GLV, and 71% overall. Completion rate of weekly counseling calls was 90%. The experimental group reported an overall mean increase of 5.5 cups GLV and 3.5 cups CV per week from baseline intake, respectively. No significant between- or within-arm differences were observed for inflammatory markers or carotenoids. CONCLUSION: A posttreatment intervention aimed at increasing CV and GLV intake in HNC survivors is feasible. A larger RCT is needed to assess the efficacy of this intervention on disease outcomes.

Early Outcomes of a Multilevel Human Papillomavirus Vaccination Pilot Intervention in Federally Qualified Health Centers.

OBJECTIVE: Human papillomavirus (HPV) vaccine coverage in the United States remains low compared with other adolescent vaccines. As the largest primary care network in the United States, safety net clinics such as federally qualified health centers (FQHCs) serve patients at a disproportionate risk of HPV-related cancers. In this pilot project, the American Cancer Society (ACS) leveraged its primary care workforce to implement quality improvement interventions in the unique context of 30 FQHC systems across the country, including 130 clinic sites reaching >20,000 adolescents in a variety of geographic settings. METHODS: FQHC systems were randomly selected to receive either a $90,000 2-year grant, a $10,000 3-month grant, or training and technical assistance without funding. All 3 intervention groups conducted provider training and education, completed a capacity assessment tool, and measured HPV vaccination rates. Annual HPV vaccine series initiation and completion rates for active, 11- to 12-year-old patients were measured to evaluate project outcomes. RESULTS: HPV vaccine series initiation rates among 11- to 12-year-old patients increased by 14.6 percentage points from a baseline of 41.2% before the intervention (2014) to the intervention year (2015). Changes in HPV second dose and series completion rates were not statistically significant. Meningococcal and tetanus, diphtheria, and acellular pertussis vaccination rates also increased significantly, by 13.9 and 9.9 percentage points from baseline rates of 49.1% and 52.5%, respectively. CONCLUSIONS: The first year of this pilot project showed early success, particularly with HPV vaccine series initiation. On the basis of these promising results, ACS is expanding clinical quality improvement projects to increase HPV vaccination across the country.

Dietary soy protein and isoflavones have no significant effect on bone and a potentially negative effect on the uterus of sexually mature intact Sprague-Dawley female rats.

OBJECTIVE: To evaluate the effect of dietary soy protein and isoflavones on bone and the reproductive tract in premenopausal rats. DESIGN: Three-month-old intact Sprague-Dawley female rats (N = 50) were fed diets containing casein, soy protein, or casein with isoflavone extract for 12 weeks. The amount of casein, soy protein, and extract (per kilogram diet) in each group was: (1) 200 g casein (control); (2) 100 g casein plus 100 g soy protein (low soy); (3) 200 g soy protein (high soy); 4) 200 g casein plus 17.2g extract (low extract); and (5) 200 g casein plus 34.4 g extract (high extract). Diet consumption, body weight, uterine wet weight, urinary deoxypyridinoline concentration, and bone mineral density of the femur and lumbar vertebrae were measured. Femur rigidity was evaluated by histomorphometry. The uterus and vagina were studied histologically. RESULTS: Rats in all treatment groups had lower body weights and lower deoxypyridinoline concentrations compared with controls, but none of the differences was statistically significant. There was no significant difference in femur and lumbar bone mineral density, uterine wet weights, or histomorphometry between the control and treatment groups. Histologically, uteri and vaginae were normal in all groups except that 1 of 10 rats in the high-soy group and 2 of 10 rats in the high-extract group showed extensive squamous metaplasia in the uterine gland. CONCLUSION: These results suggest that dietary isolated soy protein and isoflavones have no effect on bone and the vagina during premenopausal period, but may have an adverse effect on the uterus.

Dietary soy protein and isoflavones: minimal beneficial effects on bone and no effect on the reproductive tract of sexually mature ovariectomized Sprague-Dawley rats.

OBJECTIVE: The present study was conducted to determine the effects of dietary soy protein and isoflavones on bone and the reproductive tract in the absence of the ovary. DESIGN: Three-month-old Sprague-Dawley rats (n = 56) were either sham-operated or ovariectomized and then fed diets containing casein or soy protein +/- isoflavone extract for 12 weeks. The amounts of casein, soy protein, and extract (per kg diet) in each group were as follows: (1) Ovariectomy, 200 g of casein; (2) Ovariectomy+low soy, 100 g of casein + 100 g of soy protein; (3) Ovariectomy+high soy, 200 g of soy protein; (4) Ovariectomy+low extract, 200 g of casein + 17.2 g of extract; (5) Ovariectomy+high extract, 200 g of casein + 34.4 g of extract; (6) Ovary intact, 200 g of casein; (7) Ovariectomy+estradiol-17beta, 200 g of casein. Diet consumption, body weight, uterine weight, urine deoxypyridinoline, and bone mineral density of the femur and lumbar vertebrae were measured. The femur rigidity was evaluated by histomorphometry. The reproductive tract (uterus, vagina, and cervix) was studied histologically. RESULTS: The Ovariectomy group showed significant increases in body weight, diet consumption, and deoxypyridinoline, decreases in uterine weight and bone mineral density, and negative changes in histomorphometry compared with the Ovary intact group. Neither soy protein nor extract diets abrogated these alterations, except for the Ovariectomy+high extract group that showed statistically significant positive changes in histomorphometric parameters. There were no histological differences in the reproductive tract among Ovariectomy, Ovariectomy+soy, and Ovariectomy+extract groups. The estradiol-17beta replacement abrogated ovariectomy-induced alterations. CONCLUSION: Dietary intake of isoflavones by sexually mature ovariectomized rats has a minimal beneficial effect on bone with no effect on the reproductive tract.

Dietary soy protein and isoflavones: no effect on the reproductive tract and minimal positive effect on bone resorption in the intact female Fischer 344 rat.

Our previous study evaluating 3 months of feeding soy protein or isoflavones (IF) to intact adult Sprague-Dawley (SD) rats showed no change in bone and the vagina but occasional extensive squamous metaplasia of the uterine glandular epithelium was observed. The current study was designed to characterize further these effects of soy protein or IF on the uterus using the Fischer 344 (F344) rat, a known high responder strain to estrogenic stimuli. Three-month-old intact F344 rats were divided into five groups and fed diets containing either casein, low or high amount of soy protein or casein with low or high amount of IF extract. Body weight, urinary deoxypyridinoline (Dpyr), bone mineral density (BMD) of the femur and lumbar, uterine wet weight and histology of the reproductive tract were evaluated. No significant difference was seen in bone parameters between control and treatment groups except for a lower Dpyr in the high soy and a higher lumbar BMD in the low soy groups. No alteration was seen in the reproductive tract of all treatment groups. Contrary to our hypothesis, the present results suggest that the uterus of the F344 strain is less sensitive to dietary soy protein and IF than that of the SD strain.

Administering caudal anesthesia at completion of clubfoot surgery does not affect postoperative use of narcotics.

In the pediatric population, control of postoperative pain is a challenging and important issue. We conducted this retrospective study to determine whether single-dose caudal anesthesia administered after club-foot surgery helps to decrease postoperative use of narcotics. Fifty-one patients given an injection of caudal anesthesia (bupivacaine) at completion of clubfoot surgery were compared with 41 patients who did not receive a caudal block. Postoperative pain control was assessed by recording how much narcotic was used by each patient during time in the recovery room and during the first 8 hours after surgery. Results show that a single dose of caudal anesthesia administered at completion of clubfoot surgery is not associated with a statistically significant change in use of narcotics during either postoperative period.