Control of OSA in a patient with CCC of soft palate using bilateral hypoglossal nerve stimulation.

Patients with complete concentric collapse of the redropalatal airway are excluded from unilateral hypoglossal nerve stimulation. This case report shows good control of OSA in a patient with CCC with a new bilateral hypoglossal nerve stimulator.

Anthocyanins and their physiologically relevant metabolites alter the expression of IL-6 and VCAM-1 in CD40L and oxidized LDL challenged vascular endothelial cells.

SCOPE: In vitro and in vivo studies suggest that dietary anthocyanins modulate cardiovascular disease risk; however, given anthocyanins extensive metabolism, it is likely that their degradation products and conjugated metabolites are responsible for this reported bioactivity. METHODS AND RESULTS: Human vascular endothelial cells were stimulated with either oxidized LDL (oxLDL) or cluster of differentiation 40 ligand (CD40L) and cotreated with cyanidin-3-glucoside and 11 of its recently identified metabolites, at 0.1, 1, and 10 µM concentrations. Protein and gene expression of IL-6 and VCAM-1 was quantified by ELISA and RT-qPCR. In oxLDL-stimulated cells the parent anthocyanin had no effect on IL-6 production, whereas numerous anthocyanin metabolites significantly reduced IL-6 protein levels; phase II conjugates of protocatechuic acid produced the greatest effects (>75% reduction, p ≤ 0.05). In CD40L-stimulated cells the anthocyanin and its phase II metabolites reduced IL-6 protein production, where protocatechuic acid-4-sulfate induced the greatest reduction (>96% reduction, p ≤ 0.03). Similarly, the anthocyanin and its metabolites reduced VCAM-1 protein production, with ferulic acid producing the greatest effect (>65% reduction, p ≤ 0.04). CONCLUSION: These novel data provide evidence to suggest that anthocyanin metabolites are bioactive at physiologically relevant concentrations and have the potential to modulate cardiovascular disease progression by altering the expression of inflammatory mediators.

Phenolic metabolites of anthocyanins modulate mechanisms of endothelial function.

Anthocyanins are reported to have vascular bioactivity, however their mechanisms of action are largely unknown. Evidence suggests that anthocyanins modulate endothelial function, potentially by increasing nitric oxide (NO) synthesis, or enhancing NO bioavailability. This study compared the activity of cyanidin-3-glucoside, its degradation product protocatechuic acid, and phase II metabolite, vanillic acid. Production of NO and superoxide and expression of endothelial NO synthase (eNOS), NADPH oxidase (NOX), and heme oxygenase-1 (HO-1) were established in human vascular cell models. Nitric oxide levels were not modulated by the treatments, although eNOS was upregulated by cyanidin-3-glucoside, and superoxide production was decreased by both phenolic acids. Vanillic acid upregulated p22(phox) mRNA but did not alter NOX protein expression, although trends were observed for p47(phox) downregulation and HO-1 upregulation. Anthocyanin metabolites may therefore modulate vascular reactivity by inducing HO-1 and modulating NOX activity, resulting in reduced superoxide production and improved NO bioavailability.

Methods for isolating, identifying, and quantifying anthocyanin metabolites in clinical samples.

The metabolic fate of anthocyanins until recently was relatively unknown, primarily as a result of their instability at physiological pH and a lack of published methods for isolating and identifying their metabolites from biological samples. The aim of the present work was to establish methods for the extraction and quantification of anthocyanin metabolites present in urine, serum, and fecal samples. 35 commercial and 10 synthetic analytes, including both known and predicted human and microbial metabolites of anthocyanins, were obtained as reference standards. HPLC and MS/MS conditions were optimized for organic modifier, ionic modifier, mobile phase gradient, flow rate, column type, MS source, and compound dependent parameters. The impact of sorbent, solvent, acid, preservative, elution, and evaporation on solid phase extraction (SPE) efficiency was also explored. The HPLC-MS/MS method validation demonstrated acceptable linearity (R(2), 0.997 ± 0.002) and sensitivity (limits of detection (LODs): urine, 100 ± 375 nM; serum, 104 ± 358 nM; feces 138 ± 344 nM), and the final SPE methods provided recoveries of 88.3 ± 17.8% for urine, 86.5 ± 11.1% for serum, and 80.6 ± 20.9% for feces. The final methods were applied to clinical samples derived from an anthocyanin intervention study, where 36 of the 45 modeled metabolites were detected within urine, plasma, or fecal samples. The described methods provide suitable versatility for the identification and quantification of an extensive series of anthocyanin metabolites for use in future clinical studies exploring absorption, distribution, metabolism, and elimination.

Human metabolism and elimination of the anthocyanin, cyanidin-3-glucoside: a (13)C-tracer study.

BACKGROUND: Evidence suggests that the consumption of anthocyanin-rich foods beneficially affects cardiovascular health; however, the absorption, distribution, metabolism, and elimination (ADME) of anthocyanin-rich foods are relatively unknown. OBJECTIVE: We investigated the ADME of a (13)C5-labeled anthocyanin in humans. DESIGN: Eight male participants consumed 500 mg isotopically labeled cyanidin-3-glucoside (6,8,10,3',5'-(13)C5-C3G). Biological samples were collected over 48 h, and (13)C and (13)C-labeled metabolite concentrations were measured by using isotope-ratio mass spectrometry and liquid chromatography-tandem mass spectrometry. RESULTS: The mean ± SE percentage of (13)C recovered in urine, breath, and feces was 43.9 ± 25.9% (range: 15.1-99.3% across participants). The relative bioavailability was 12.38 ± 1.38% (5.37 ± 0.67% excreted in urine and 6.91 ± 1.59% in breath). Maximum rates of (13)C elimination were achieved 30 min after ingestion (32.53 ± 14.24 µg(13)C/h), whereas (13)C-labeled metabolites peaked (maximum serum concentration: 5.97 ± 2.14 µmol/L) at 10.25 ± 4.14 h. The half-life for (13)C-labeled metabolites ranged between 12.44 ± 4.22 and 51.62 ± 22.55 h. (13)C elimination was greatest between 0 and 1 h for urine (90.30 ± 15.28 µg/h), at 6 h for breath (132.87 ± 32.23 µg/h), and between 6 and 24 h for feces (557.28 ± 247.88 µg/h), whereas the highest concentrations of (13)C-labeled metabolites were identified in urine (10.77 ± 4.52 µmol/L) and fecal samples (43.16 ± 18.00 µmol/L) collected between 6 and 24 h. Metabolites were identified as degradation products, phenolic, hippuric, phenylacetic, and phenylpropenoic acids. CONCLUSION: Anthocyanins are more bioavailable than previously perceived, and their metabolites are present in the circulation for ≤48 h after ingestion. This trial was registered at clinicaltrials.gov as NCT01106729.

Simultaneous pasteurization and homogenization of human milk by combining heat and ultrasound: effect on milk quality.

The combination of ultrasound and heat (thermoultrasound) is an emerging food preservation technique that retains higher quantities of bioactive components compared with current thermal pasteurization practice, but has not yet been assessed for pasteurizing human milk. Artificially contaminated human milk samples were treated with ultrasound (20 kHz, 150 watts) with and without heating. The retention of four human milk proteins was quantified by biochemical assay and laser scattering particle sizing was used to determine the extent of homogenization. While ultrasonic treatment was effective at inactivating Escherichia coli (D4 degrees C=5.94 min), Staphylococcus epidermidis exhibited resistance (D4 degrees C=16.01 min). Thermoultrasonic treatment was considerably more effective (Esch. coli D45 degrees C=1.74 min, D50 degrees C=0.89 min; Staph. epidermidis D45 degrees C=2.08 min, D50 degrees C=0.94 minutes) with a predicted retention (2.8 min treatment, 50 degrees C) of secretory IgA lysozyme, lactoferrin and bile salt stimulated lipase of 91, 80, 77, and 45%, respectively. Homogenization of the milk samples occurred after 5 min and 2 min of ultrasonic and thermoultrasonic treatment, respectively. Thermoultrasonic treatment is an effective method for pasteurizing donor human milk and retaining a greater proportion of bioactive components compared with current practices. However, further studies are required to assess the practicality of applying this technique routinely to donor human milk.

Design and characterization of a human milk product for the preterm infant.

BACKGROUND: It is necessary to fortify human milk to promote optimal growth of the very preterm infant. However, the addition of non-human milk components to human milk is not ideal because of the risk of feeding intolerance and necrotizing enterocolitis. Human milk products (HMP) are an alternative to commercially available fortifiers, but their preparation is likely to result in modifications to the qualities of human milk. METHODS: Ten batches of HMP were prepared with the aim of meeting a desired protein:energy ratio of 3.0 g of protein/100 kcal. Ultrafiltration was used to produce a skim milk concentrate, to which cream was then added to produce the final HMP. Characterization of HMP and human milk fortified with commercial human milk fortifiers (Nutriprem [Cow & Gate, Limerick, Ireland] and S-26 SMA human milk fortifier [Wyeth Nutrition, Baulkham Hills, NSW, Australia]) included quantifying macronutrient content, osmolality, microbial content, and particle distribution. RESULTS: Average protein:energy ratio of the final batch was 2.93 +/- 0.10 g of protein/100 kcal, equating to an inaccuracy of 2.5% relative to the desired ratio of 3.0 g of protein/100 kcal. Significantly greater fat (P < 0.01), lower lactose (P < 0.001), and lower osmolality (P < 0.001) were characteristic of the HMP compared to human milk fortified with either commercial fortifier. Microbial growth occurred during preparation of HMP but did not exceed 10(5) colony-forming units/mL, and pasteurization of human milk prevented contamination in 80% of batches. CONCLUSIONS: HMP can be designed to accurately target the protein and energy requirements of the preterm infant, but modifications of the macronutrient, biochemical, structural, and microbial characteristics of human milk may affect the quality of the final product.

Retention of the immunological proteins of pasteurized human milk in relation to pasteurizer design and practice.

Pasteurizing donor human milk inactivates bacteria that may be of concern to the preterm infant. However, current practice for Holder Pasteurization (62.5 degrees C for 30 min) is detrimental to the bioactivity of human milk. An experimental pasteurizer was used to determine the maximum temperature at which 90% of secretory IgA, lysozyme, and lactoferrin were retained and whether this temperature was capable of inactivating five common bacterial contaminants. The retention of these proteins was also compared using a commercially available bottle immersion or holding chamber system. After pasteurization at 62.5 degrees C for 30 min, the retention across all three systems was 72.3 +/- 3.6%, 21.8 +/- 3.3%, and 39.4 +/- 11.5% for sIgA, lactoferrin, and lysozyme, respectively (n = 22). The retention of all three proteins was at least 90% when human milk was pasteurized at 57 degrees C for 30 min, and this temperature was also effective at removing 99.9% of all inoculated bacterial species. In addition, human milk that was pasteurized in the experimental system had a significantly higher proportion of lysozyme compared with samples pasteurized in the bottle immersion system. These findings suggest that optimizing pasteurization temperature and improving pasteurizer design enhances the quality of pasteurized donor human milk.

A method for standardizing the fat content of human milk for use in the neonatal intensive care unit.

BACKGROUND: Accurately targeting the nutritional needs of the early preterm infant is challenging when human milk is used due to the natural variation in energy composition. The purpose of this study was to develop and evaluate a simple method for reducing the variation in fat and energy content of human milk prior to fortification such that the infant receives a diet of known composition. METHODS: Milk was centrifuged at low speed to concentrate the fat into a cream layer and a predetermined volume of skim milk is removed to meet a specific fat concentration. The fat layer is then resuspended to produce reconstituted milk of a specified standard fat content. RESULTS: Using this method it was possible to reduce the coefficient of variation in fat content of six different samples of donor human milk from 19.3% to 2.6%. As fat globule size may be associated with fat absorption, the effect that centrifugation and resuspension had on human milk fat globule distribution was assessed by laser diffraction particle sizing. No difference in the particle distribution of the treated and untreated human milk was observed. CONCLUSION: This method is accurate and simple, allowing for integration alongside current milk bank and NICU practices for use with both donor human milk and mother's own milk.