Improvements in extraction yield by solid phase lipid extraction from liquid infant formula and human milk, and the fatty acid distribution in milk TAG analyzed by joint JOCS/AOCS official method Ch 3a-19.

The Röse-Gottlieb method is one of the most widely used methods for extracting lipids from milk samples. However, we found that lipid recovery from liquid infant formula and human breast milk was lower than expected. Better lipid recovery from these liquid matrices was obtained by solid phase extraction using silica gel; ~10% more could be recovered from liquid infant formula and ruminant milk, and 25% more from human breast milk. However, the method is not recommended for lipid extraction from dried whole milk powders.

Comprehensive Analysis of Lipid Composition in Human Foremilk and Hindmilk.

Precision nutrition, also referred to as personalized nutrition, focuses on the individual to determine the individual's most effective eating plan to prevent or treat disease. A precision nutrition for infants requires the determination of the profile of human milk. We compared the lipid profiles of the foremilk (i.e., the initial milk of a breastfeed) and hindmilk (the last milk) of six Japanese subjects and evaluated whether a human milk lipid profile is useful for precision nutrition even though the fat concentration fluctuates during lactation. We detected and quantified 527 species with a lipidome analysis by liquid chromatography-tandem mass spectrometry. The fat concentration in hindmilk (120.6 ± 66.7 µmol/mL) was significantly higher than that in foremilk (68.6 ± 33.3 µmol/mL). While the total carbon number of fatty acids in triglyceride (TG) was highest in C52 for all subjects, the second or third number differed among the subjects. Both the distribution of total carbon number of fatty acids included in TG and the distribution of fatty acids in TG classified by the number of double bonds were almost the same in the foremilk and hindmilk in each subject. The lipids levels containing docosahexaenoic acid and arachidonic acid in total lipids of the foremilk and the hindmilk were almost the same in each subject. Among the sphingolipids and glycerophospholipids, the level of sphingomyelin was the highest in four subjects' milk, and phosphatidylcholine was the highest in the other two subjects' milk. The order of their major species was the same in each foremilk and hindmilk. A clustering heatmap revealed the differences between foremilk and hindmilk in the same subject were smaller than the differences among individuals. Our analyses indicate that a human-milk lipid profile reflects individual characteristics and is a worthwhile focus for precision nutrition.

Effects of Oral Supplementation with Paprika Xanthophylls on Human Skin Moisture.

Carotenoids, classified into carotenes and xanthophylls, are natural lipophilic pigments that are widely distributed in plants. Red paprika is unique in its high levels of various xanthophylls. Dietary paprika xanthophylls have been shown to reduce UV-induced photo damage by the strong antioxidant activity in the skin. However, the precise effects of paprika xanthophylls on skin condition are still unknown. Here we show that skin moisture is enhanced by the intake of red paprika supplements including seven xanthophylls. We conducted a 4-week randomized, single-blind, parallel-group controlled trial to clarify the effects of dietary paprika xanthophylls on facial skin. The results showed that the moisture was significantly higher in the paprika intake group than in the control (21.0±8.9 vs 13.4±6.0 (A.U.)). There was no significant difference between the paprika and control groups in other parameters such as viscoelasticity, the number of wrinkles, and the amount of water evaporation. On the other hand, the number of brown stains in the paprika group increased significantly, to 190±26 from 173±30 (p < 0.05). In vitro experiments, quantitative real-time PCR showed that paprika extract led to increases in the gene expression of Aquaporin 3 (AQP3) and hyaluronic acid synthase 3 (HAS3) in cultured keratinocytes. Western blotting showed that the paprika extract enhanced AQP3 expression. Taken together, taking supplements containing paprika xanthophylls may provide beneficial effects of moisture on facial skin. The study provides new insights into understanding the role of paprika xanthophylls in the skin.

Analysis of Fatty Acid Esters of Hydroxyl Fatty Acid in Nut Oils and Other Plant Oils.

Oils and lipids are common food components and efficient sources of energy. Both the quantity and the quality of oils and lipids are important with regard to health and disease. Fatty acid ester of hydroxy fatty acid (FAHFA) is a novel lipid class that was discovered as an endogenous lipid; FAHFAs have shown anti-diabetic effects in a mammalian system. We analyzed the overall FAHFA composition in nut oils and other common oils: almond (raw, roasted), walnut, peanut, olive, palm, soybean, and rapeseed oils. We developed a method of liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC-ESI/MS/MS) for a comprehensive target analysis of FAHFAs. The analysis revealed wide variation in the FAHFA profiles (15 compounds and 62 peaks). For 7-11 compounds of FAHFA, a total level of 8-29 pmol/mg oil was detected in nuts oils; for 11 compounds, 4.9 pmol/mg oil was detected in olive oil, and for 4-9 compounds, < 2 pmol/mg oil was detected in palm, soy, and rapeseed oils. The major FAHFAs were FAHFA 36:3, FAHFA 36:2, and FAHFA 36:4 in nut oil, FAHFA 36:2, FAHFA 34:1, and FAHFA 36:1 in olive oil, and FAHFA 32:1, FAHFA 34:0, FAHFA 36:0, and FAHFA 36:1 in all of the common oils. The composition of FAHFAs in nut oils is mainly unsaturated fatty acids, whereas those in olive oil are unsaturated fatty acids and saturated fatty acids. The composition of FAHFAs in common oils was mainly saturated fats. This is the first report to demonstrate the quality and quantity of the FAHFAs in the nut oils. Nuts have been described to be a great source of many nutrients and to be beneficial for our health. Our present findings comprise additional evidence that the intake of nuts in daily diets may prevent metabolic and inflammatory-based diseases.

Corrigendum: Gravity-Induced Lower-Leg Swelling Can Be Ameliorated by Ingestion of α-Glucosyl Hesperidin Beverage.

[This corrects the article DOI: 10.3389/fphys.2021.670640.].

Gravity-Induced Lower-Leg Swelling Can Be Ameliorated by Ingestion of α-Glucosyl Hesperidin Beverage.

The most likely cause of lower-leg swelling is prolonged sitting, which sometimes induces deep vein thrombosis, also known as, economy class syndrome. We aimed to clarify the influence of intake of 4 G -α-glucopyranosyl hesperidin (G-Hsp) beverage on the lower-leg swelling caused by 6 h of sitting in six healthy women. All subjects ingested 100 mL of G-Hsp or Placebo beverages with 100 mL of mineral water after 10 min of rest in a chair. Subsequently, subjects were requested to sit in the chair in a relaxed position for 6 h with two breaks to walk for urination. Calf water content measured by impedance plethysmography, calf circumference, and calf skin temperature by infrared thermography were measured, along with assessment of calf swelling sensation on a visual analog scale. Increase in ankle % circumference was significantly less after the G-Hsp ingestion (101.8 ± 1.5%) than after placebo (103.3 ± 0.8%; P = 0.004). A significant difference was found between percent circumference after the G-Hsp and the placebo, that is, the calf swelling after the placebo was significantly larger (P = 0.043). A gradual increase in skin temperature at the lower limb was observed after G-Hsp ingestion, while there was no change after placebo. Gravity-induced calf and ankle swelling resulted by prolonged sitting can be ameliorated by oral ingestion of hesperidin-derived G-Hsp through production of nitric oxide. It might be helpful in preventing economy-class syndrome caused by enforced sitting for a long duration.

Inhibition of Atherosclerotic Plaque Development by Oral Administration of α-Glucosyl Hesperidin and Water-Dispersible Hesperetin in Apolipoprotein E Knockout Mice.

OBJECTIVE: Hesperidin, an abundant flavonoid in citrus fruit, and its aglycone, hesperetin, have been reported to possess various physiological activities, including antioxidant, anti-inflammatory, hypolipidemic, and antihypertensive activities. In this study, we investigated whether α-glucosyl hesperidin and water-dispersible hesperetin have protective effects on atherosclerotic progression in apolipoprotein E knockout (Apo-E KO) mice. METHODS: Ten-week-old male Apo-E KO mice were randomly assigned a regular high-fat diet, a high-fat diet with 0.5% α-glucosyl hesperidin, or a high-fat diet with 0.1% water-dispersible hesperetin for 12 weeks. Measurement of plasma total cholesterol levels, histological staining of aortic root, and immunohistochemistry for macrophages were performed to evaluate atherosclerotic plaque formation. Vascular reactivity of mouse aortic rings was also measured. RESULTS: Both α-glucosyl hesperidin and water-dispersible hesperetin reduced plasma total cholesterol level. They also reduced plaque formation area, adipose deposition, and macrophage infiltration into atherosclerotic lesion. Vascular-endothelium-dependent relaxation in response to acetylcholine was improved in both experimental diet groups compared to the high-fat diet group. CONCLUSIONS: Our study suggests that both α-glucosyl hesperidin and water-dispersible hesperetin exert protective effects on atherosclerotic progression in Apo-E KO mice because they exhibit hypolipidemic activity, reduce inflammation through macrophages, and prevent endothelial dysfunction.

Qualitative and Quantitative Analyses of Glycogen in Human Milk.

Identification as well as a detailed analysis of glycogen in human milk has not been shown yet. The present study confirmed that glycogen is contained in human milk by qualitative and quantitative analyses. High-performance anion exchange chromatography (HPAEC) and high-performance size exclusion chromatography with a multiangle laser light scattering detector (HPSEC-MALLS) were used for qualitative analysis of glycogen in human milk. Quantitative analysis was carried out by using samples obtained from the individual milks. The result revealed that the concentration of human milk glycogen varied depending on the mother's condition-such as the period postpartum and inflammation. The amounts of glycogen in human milk collected at 0 and 1-2 months postpartum were higher than in milk collected at 3-14 months postpartum. In the milk from mothers with severe mastitis, the concentration of glycogen was about 40 times higher than that in normal milk.

Neuroprotective effect of water-dispersible hesperetin in retinal ischemia reperfusion injury.

PURPOSE: To determine whether water-dispersible hesperetin (WD-Hpt) can prevent degeneration of ganglion cell neurons in the ischemic retina. METHODS: Ischemia reperfusion (I/R) injury was induced by increasing the intraocular pressure of mice to 110 mmHg for 40 min. Mice received daily intraperitoneal injections with either normal saline (NS, 0.3 ml/day) or WD-Hpt (0.3 ml, 200 mg/kg/day). Reactive oxygen species (ROS) was assessed by dihydroethidium and nitrotyrosine formation. Inflammation was estimated by microglial morphology in the retina. Lipopolysaccharide (LPS)-stimulated BV-2 cells were used to explore the anti-inflammatory effect of WD-Hpt on activated microglia by quantifying the expression of IL-1ß using real-time quantitative reverse transcription-polymerase chain reaction. Ganglion cell loss was assessed by immunohistochemistry of NeuN. Glial activation was quantified with glial fibrillary acidic protein (GFAP) immunoreactivity. Apoptosis was evaluated with a terminal deoxynucleotidyl transferase (TUNEL) assay and immunohistochemistry of cleaved caspase-3. Phosphorylation of extracellular signal-regulated kinase (p-ERK) was surveyed by western blotting. RESULTS: WD-Hpt decreased I/R-induced ROS formation. WD-Hpt alleviated microglial activation induced by I/R and reduced mRNA levels of IL-1ß in LPS-stimulated BV-2. I/R resulted in a 37% reduction in the number of ganglion cells in the NS-treated mice, whereas the reduction was only 5% in the WD-Hpt-treated mice. In addition, WD-Hpt mitigated the immunoreactivity of GFAP, increased expression of cleaved caspase-3, increased number of TUNEL positive cells and p-ERK after I/R. CONCLUSIONS: WD-Hpt protected ganglion cells from I/R injury by inhibiting oxidative stress and modulating cell death signaling. Moreover, WD-Hpt had an anti-inflammatory effect through the suppression of activated microglia.

Bioavailability of orally administered water-dispersible hesperetin and its effect on peripheral vasodilatation in human subjects: implication of endothelial functions of plasma conjugated metabolites.

Hesperetin is an aglycone of citrus flavonoids and is expected to exert a vasodilatation effect in vivo. We developed water-dispersible hesperetin by the process of micronization to enhance the bioavailability of hesperetin. This study aimed to assess the effect of this process on the bioavailability of hesperetin and to estimate its efficiency on vasodilatation-related functions using endothelial cells in vitro and a human volunteer study at a single dose in vivo. We found that water-dispersible hesperetin was absorbed rapidly, with its maximum plasma concentration (C(max)) being 10.2 ± 1.2 µM, and that the time to reach C(max), which is within 1 h if 150 mg of this preparation was orally administered in humans. LC-MS analyses of the plasma at C(max) demonstrated that hesperetin accumulated in the plasma as hesperetin 7-O-ß-D-glucuronide (Hp7GA), hesperetin 3'-O-ß-D-glucuronide (Hp3'GA) and hesperetin sulfate exclusively. Similar to hesperetin, Hp7GA enhanced nitric oxide (NO) release by inhibiting nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase) activity in a human umbilical vein endothelial cell culture system, indicating that plasma hesperetin metabolites can improve vasodilatation in the vascular system. A volunteer study using women with cold sensitivity showed that a single dose of water-dispersible hesperetin was effective on peripheral vasodilatation.These results strongly suggest that rapid accumulation with higher plasma concentration enables hesperetin to exert a potential vasodilatation effect by the endothelial action of its plasma metabolites. Water-dispersible hesperetin may be useful to improve the health effect of dietary hesperetin.

Effects of long-term feeding of α-glucosylhesperidin on the mechanical properties of rabbit femoral arteries.

Many people are sensitive to cold, resulting in poor blood circulation. There is evidence that hesperidin results in increased peripheral circulation and skin temperature. A transglycosylated hesperidin, α-glucosylhesperidin, is more bioabsorbable than hesperidin. In the present study, biomechanical studies were performed on the effects of long-term feeding of α-glucosylhesperidin on the contractile response (diameter response) and stiffness of femoral arteries excised from rabbits. Animals in the normal (non-treated), low, and high groups were fed 0, 150 and 4500 mg/day, respectively, of α-glucosylhesperidin for about 24 weeks. The feeding of α-glucosylhesperidin did not change arterial stiffness nor mean blood flow rate in the femoral artery; however, it increased mean aortic blood pressure and decreased arterial diameter at 100 mmHg in the high group. The diameter responses developed by 10-5 M of norepinephrine were significantly lower in the high and low groups than in non-treated group. This result indicates that, due to the long-term feeding of α-glucosylhesperidin, arterial contraction induced by the neurotransmitter of sympathetic nerves decreases. It was estimated that blood flow in such muscular arteries as the femoral artery is maintained at normal by α-glucosylhesperidin even under the conditions of autonomic imbalance and cold intolerance.

Tissue distribution of hesperetin in rats after a dietary intake.

Hesperetin, the aglycone of hesperidin present in citrus fruits, possesses various biological activities. We assessed the tissue distribution of hesperetin in rats fed with a 0.2% hesperetin diet for 4 weeks. Its highest concentration was found in the liver, and the second highest was in the aorta. The aorta is assumed to be one of the main target tissues of hesperetin for exerting its functions.

Effects of alpha-glucosylhesperidin on the peripheral body temperature and autonomic nervous system.

We studied the effects of alpha-glucosylhesperidin (G-Hsp) on the peripheral body temperature and autonomic nervous system in humans. We first conducted a survey of 97 female university students about excessive sensitivity to the cold; 74% of them replied that they were susceptible or somewhat susceptible to the cold. We subsequently conducted a three-step experiment. In the first experiment, G-Hsp (500 mg) was proven to prevent a decrease in the peripheral body temperature under an ambient temperature of 24 degrees C. In the second experiment, a warm beverage containing G-Hsp promoted blood circulation and kept the finger temperature higher for a longer time. We finally used a heart-rate variability analysis to study whether G-Hsp changed the autonomic nervous activity. The high-frequency (HF) component tended to be higher, while the ratio of the low-frequency (LF)/HF components tended to be lower after the G-Hsp administration. These results suggest that the mechanism for temperature control by G-Hsp might involve an effect on the autonomic nervous system.

Effect of 4G-alpha-glucopyranosyl hesperidin on brown fat adipose tissue- and cutaneous-sympathetic nerve activity and peripheral body temperature.

Changes in the activity of the autonomic nervous system are good indicators of alterations in physiological phenomena such as the body temperature, blood glucose, blood pressure. Hesperidin, a flavanone known as vitamin P, has been shown to reduce the levels of serum lipids, cholesterol, and blood pressure. However, hesperidin is not water-soluble and is not well absorbed from the intestine. G-hesperidin (4G-alpha-glucopyranosyl hesperidin) is more water-soluble and more rapidly absorbed than hesperidin. In order to clarify the functions of G-hesperidin, we examined the effects of oral administration of G-hesperidin on interscapular brown adipose tissue-sympathetic nerve activity (BAT-SNA) and cutaneous sympathetic nerve activity (CASNA) in rats weighing about 300 g. In this study, we found that oral administration of 60 mg of G-hesperidin increased the BAT-SNA but decreased the CASNA in urethane-anesthetized rats. Since an elevation in BAT-SNA increases heat production (i.e. body temperature (BT)) and a decrease in CASNA increases cutaneous perfusion, we examined whether oral administration of G-hesperidin had an effect on the peripheral BT in rats. Consequently, we observed that the subcutaneous BT at the caudal end of the back after oral administration of 60 mg of G-hesperidin was significantly higher than the subcutaneous BT after oral administration of water in conscious rats. These findings suggest that G-hesperidin enhances the BAT-SNA and suppresses the CASNA resulting in an increase in the peripheral BT, probably by an increase in the thermogenesis in the BAT and an elevation in the cutaneous blood flow.