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The relationship between the softening of yuzu and the extraction of pectin during soaking in citric acid, heating or pressurization was investigated. Firmness of peel was greatest to least; pressurized > soaked for 24 hrs at pH 2.7 > heated, respectively. The cell walls did not loosen after pressurization. However, after heating, the middle lamella of albedo separated but flavedo did not. The amount of pectin was greatest to least in albedo > flavedo > segment walls > juice sacs, and in flavedo, raw > pressurized >
heated > soaked at pH 2.7 > boiled flavedo. About 24% of pectin was extracted by soaking at pH 2.7. By soaking, peel softened due to the removal of Ca2+. The pH values of peel and cooked water were 3 - 4 and 4.02, respectively. Therefore, it was found that degradation of pectin by heating was mainly from hydrolysis and not by trans-elimination.
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Lysophosphatidic acid (LPA) is a lipid mediator involved in various physiological responses, including wound healing. Evidence of the antiulcer activity of LPA has been reported, and soybean LPA at concentration of 10 M is effective to reduce stress-induced gastric ulcer. Since LPA can be formed from phosphatidic acid (PA) by digestive phospholipase A2, dietary PA can be considered a potential antiulcer phospholipid. In this study, PA production in cut processing of cabbage leaves was examined. The amounts of PA in sliced, minced and homogenized cabbage leaves were
107±5, 134±19 and 286±29 nmol PA/g (wet weight), respectively, all being significantly higher than the amount of PA found in intact leaves. Mixing mayonnaise with sliced cabbage dramatically increased the PA content (1586±393 nmol/3g), indicating phospholipase D activity leaked raw cabbage produced PA. These results indicate that fine cutting raw cabbage leaves and mixing them with foods rich in phospholipids resulted in an abundant production of PA.
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Apical application of lysophosphatidic acid (LPA), a growth-factor-like phospholipid, was shown to prevent or restore gastrointestinal (GI) disorders, such as diarrhea and stomach ulcer, in experimental animals. Because LPA is formed from phosphatidic acid (PA) by the activity of digestive phospholipase A2, PA is a potential component for dietary treatment of such GI disorders. Here, we quantified PA contained in 38 foodstuffs and 3 herbs by a thin-layer-chromatography-imaging technique. Vegetables belonging to Brassicaceae, such as cabbage leaves (700 nmol/g of wet weight) and Japanese radish leaves (570 nmol/g), contained higher amounts of PA than other foodstuffs. Amounts of PA in fruits, cereals, and starchy root vegetables were below 300 nmol/g. Animal foodstuffs contained low amounts of PA (<60 nmol/g). Interestingly, leaves of Mallotus japonicas, a Japanese edible herb used for treatment of stomach ulcer, had the highest PA (1410 nmol/g) among those examined. The data shown here will be useful for the development of dietary treatment for a damaged GI tract.
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Here we examined the properties of endosperm starches and the physical properties of
cooked rice from four cultivars of Japanese upland rice containing M-type amylopectin (Chikanarijyun1, Kairyo13, Mogamichikanari1, and Hokkaiakage); S-type Nipponbare and L-type Kasalath served as standards. The amylopectin chain ratio (ACR ; ratio of the short chains of DP 10 to the short and intermediate chains of DP 24) of M-type amylopectin was higher than L-type amylopectin, and lower than S-type amylopectin.
The degree of disintegration of rice grains in 5M urea solution was in the order Nipponbare > M-type amylopectin > Kasalath. Specifically, the disintegration score of rice grains decreased as the ACR decreased. The apparent amylose (AAM) contents were in the order Kasalath > M-type amylopectin, except Kairyo13 > Nipponbare > Kairyo13.
These results indicated that there was no relationship between the AAM contents and ACR. The hardness of freshly cooked and stale rice was in the order Kasalath > M-type amylopectin > Nipponbare. The stickiness of freshly cooked rice was in the order Nipponbare > Kairyo13 > M-type amylopectin, except Kairyo13 > Kasalath and that of stale rice was in the order Nipponbare > M-type amylopectin > Kasalath. The stickiness of freshly cooked rice of Kairyo13 was the highest in the M-type amylopectin cultivars, because it had the lowest AAM content in all cultivars. Freshly cooked rice of Kairyo13 was harder and less sticky compared with that of Nipponbare. It was possible that the ACR of Kairyo13 was lower than that of Nipponbare.
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Fine structure, granule size distribution, thermal and rheological properties of starch separated from twenty kidney bean lines were determined. Apparent amylose content (AAC), intermediate fraction (mixture of relatively short amylose and long side chains of amylopectin), long-side chains of amylopectin (AP) and short side chains of amylopectin (AP) of starches were measured after debranching with isoamylase and fractionating using gel permeation chromatography. Starches from different lines showed unimodal, bimodal and trimodal distribution profiles for the granules size, majority of starches showed bimodal profile. The granule size ranged between 0.4 and 103 μm, however, granules of 10 to 30 μm were present in the highest proportion. Paste viscosities (Peak-, breakdown- and final-viscosity) had negative relation with AAC and positive with short
side chains of AP. Starches with high proportion of granules of size between 1 to 10 μm and N10 to 30 μm size, respectively had lower and higher paste viscosities. Starches with higher proportion of long side chains of AP had higher gelatinization temperature (To, Tp, Tc) and enthalpy. Starches with higher proportion of granules of size N10 to 30 μm had lower AAC and higher To, Tp and Tc. Gel hardness, To, Tp and Tc were lower for starches with higher proportion of granules of size N1 to 10 μm. Viscoelasticity of cooked starch varies with AAC, short side chains of AP and granules size distribution.
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Background: PUFA are susceptible to lipid peroxidation and play a role in inflammation, both of which can induce oxidative stress. However, the relation of PUFA to oxidative DNA damage in humans is elusive.
Objective: We examined the association between circulatory PUFA and urinary 8-oxo-7,8-dihydroguanine (8-oxoGua) concentrations in Japanese men and women.
Design: The subjects were 495 participants (290 men and 205 women) of a cross-sectional study in two municipal offices in Japan. Serum cholesterol ester (CE) and phospholipid fatty acid composition were measured by gas-liquid chromatography.
Urinary 8-oxoGua concentrations were measured by HPLC. Means of 8-oxoGua for each tertile of PUFA after adjustment for covariates were calculated by multiple regression.
Results: Urinary 8-oxoGua concentrations increased with increasing levels of n-3 PUFA, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) in serum CE (P for trend =0.001, 0.01, and 0.009, respectively), whereas they decreased with increasing levels of n-6 PUFA and linoleic acid (P for trend =0.02 and 0.051, respectively).
Conclusion: Oxidative DNA damage may be increased at higher levels of long-chain n-3 PUFA but decreased at higher levels of n-6 PUFA.
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The association of fatty acid composition with insulin resistance and type 2 diabetes has been reported in Western populations, but there is limited evidence of this association among the Japanese, whose populace consume large amounts of fish. To test the hypothesis that high palmitic, palmitoleic, and dihomo--linolenic acids and lowlevels of linoleic and n-3 fatty acids are associatedwith higher insulin resistance among the Japanese, the authors investigated the relationship between serumfatty acid composition and serumC-peptide concentrations in 437 Japanese employees aged 21 to 67 years who participated in a workplace health examination. Serumcholesterol ester and phospholipid fatty acid compositionsweremeasured by gas-liquid chromatography.Desaturase activitywas estimated by fatty acid product-to-precursor ratios.A multiple regression was used to assess the association between fatty acid and C-peptide concentrations. C-peptide concentrations were associated inversely with linoleic acid levels in
cholesterol ester and phospholipid (P for trend = .01 and .02, respectively) and positively with stearic and palmitoleic acids in cholesterol ester (P for trend =.02 and .006, respectively) and dihomo--linolenic acid in cholesterol ester and phospholipid (P for trend < .0001 for both). C-peptide concentrations were not associated with n-3 polyunsaturated fatty acids. C-peptide concentrations significantly increased as -9-desaturase (16:1 n-7/16:0) and -6-desaturase (18:3 n-6/18:2 n-6) increased (P for trend = .01 and .03, respectively) and -5-desaturase (20:4 n-6/20:3 n-6) decreased (P for trend = .004). In conclusion, a fatty acid pattern with high levels of serum stearic,palmitoleic, ordihomo--linolenic acids; -9-desaturase (16:1 n-7/16:0) or -6-desaturase (18:3 n-6/18:2n-6) activities; and lowlevels of serumlinoleic acid or -5-desaturase (20:4 n-6/20:3 n-6) activity might be associated with higher insulin resistance in Japanese adults.
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Objective: It remains unsettled whether dietary patterns play a role in insulin resistance.
Weassessed the association of major dietary patterns with C-peptide concentrations in a Japanese working population.
Methods: A cross-sectional study was conducted in 456 municipal employees (270 men and 186 women) 21 to 67 y old who participated in a health survey at the time of their periodic checkup. The dietary patterns were derived by using the principal component analysis of the consumption
of 52 food and beverage items, which was assessed by a validated brief dietary history questionnaire. Multiple regression analysis was used to estimate the means of C-peptide concentrations across tertiles of each dietary pattern score with the adjustment of potential confounders, including age, body mass index, physical activity, smoking, alcohol drinking, and energy intake.
Results: We identified three dietary patterns: healthy, animal food, and Westernized breakfast patterns. The Westernized breakfast pattern was characterized by high intakes of bread, confectionaries, and milk and yogurt but low intakes of rice and alcohol and was inversely associated with C-peptide concentrations in women but not in men. The multivariable-adjusted means of C-peptide concentrations were 1.03 ng/mL (95%
confidence interval 0.95–1.12), 0.95 ng/mL (95% confidence interval 0.88–1.03), and 0.89 ng/mL (95% confidence interval 0.82–0.97) for the lowest through the highest tertiles of the Westernized breakfast pattern score (P for trend 0.015) in women. Other dietary patterns were not appreciably associated with C-peptide concentrations. In a subgroup, similar associations were observed between dietary patterns and the homeostasis model assessment of insulin resistance.
Conclusion: The Westernized breakfast pattern may be associated with a lower insulin resistance in Japanese women.
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The content of a crude precipitate formed by creaming, which was made from a catechin mixture and caffeine, was investigated by an integral volume of H-2 proton signals of tea
catechins in the 1H-NMR spectrum. Gallated catechins formed a crude precipitate more predominantly than non-gallated catechins. The 2,3-cis-non-gallated catechin (-)-epicatechin (EC) formed a 1:1 complex with caffeine, and 2,3-cis-gallated catechin (-)epicatechin gallate (ECg) formed a 2:4 complex with caffeine. The p- p complexation site of EC with caffeine was only the A ring, whereas that of ECg included all aromatic rings, A, B, and B’. It was thought that the hydrophobicity of the 2:4 complex of ECg and caffeine was stronger than that of the 1:1 complex of EC and caffeine, with the result that the 2:4 complex of ECg and caffeine precipitated by creaming more predominantly than the 1:1 complex of EC and caffeine in aqueous solution.
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The effects of temperature and time of soaking in distilled water or tap water on the prevention of excessive softening of potatoes during cooking were investigated through changes in texture and histological structure. Potatoes were soaked in distilled water or two kinds of tap water (A and B) for 24 hrs at 20˚C, 30˚C or 60˚C, then cooked. The amount of calcium in A and B was determined. Then, calcium carbonate was added to distilled water and A to an equal concentration of B, cooked in this water. Also, potatoes were cooked at pH 6.0 ~ 8.0. Changes in texture and structure of cooked potatoes were measured by a rheoner and cryo-scanning electron microscope, respectively. Potatoes became firmer according to a longer soaking time and higher soaking temperature. As the pH value was higher, potatoes became softer. However, potatoes soaked-then-cooked in B (pH 8) were firmer than that soaked-then-cooked in A (pH 6), although the pH of B was higher than A or distilled water. The calcium concentration of B was higher than A.
When Ca2+ was added to A, the firmness of cooked potatoes became the same. Also, scanning electron microscopy showed the middle lamella separated when cooked in distilled water but not when soaked-then-cooked. The pectic substances in tissues de-esterified by soaking and preheating were resistant to degradation by trans-elimination and increased the binding with calcium; consequently, tissues remained firmer. If firmness is desired, preheating should be effective.
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Objectives of this study were to research the relationship between pectin and the softening of apples by soaking in citric acid solution, pressurizing or heating, then comparing two processing methods, high-pressure-induced and heat-induced apple jam. Apples were diced into 1cm pieces and soaked in citric acid solution (pH 2.5) for 24 hrs at 35°C or boiled for 10 min. Also, the vacuum-packed pieces were pressurized for 30 min at 500 MPa or boiled for 10 min, then, texture, structure and pectin were measured. Five kinds of apple jam were produced. Diced-apples were soaked in citric acid solution, mixed in a ratio of 1:1 or 0:1 with homogenized apples and sucrose was added (final sugar 60% at pH 2.5). These were vacuum-packed, then pressurized for 30 min at 500 MPa or boiled for 10 min, respectively. Also, a sample mixed in a ratio of 1:1 was heated with sugar and concentrated up to 60% sugar. Steady-flow viscosity, thixotropy and dynamic-viscoelasticity of jam were then measured. Sensory evaluation of jam was compared using a five point scale. Firmness of the apple decreased and cell separations were greatly when boiled > packed-then-boiled > soaked > pressurized, respectively. The amount of pectin decreased due to the removal of Ca2+ by soaking, and decreased through hydrolysis by heating but only slightly decreased by pressurization. Thus, the viscosity of concentrated jam was higher than pressure-induced and heat-induced jam. Fresh flavor, color and total evaluation of pressure-induced-jam were better than the heat-induced-jam.
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Hyuga-natsu is a typical Japanese citrus with desirous smell and edible albedo. The objectives of this study are to establish a process for pressure-induced marmalade,
compare it with heat-induced marmalade, and to investigate the softening of peel during soaking in citric acid, heating or pressurization. Albedo, segment wall and juice sacs of hyuga-natsu were homogenized with citric acid solution (pH 2.7) and mixed with sliced flavedo. Then it was soaked for 24 hrs at pH 2.7. Sucrose was then added (final sugar 50%), vacuum packed, then pressurized for 30 min at 500 MPa or boiled for 10 min, respectively. The firmness of flavedo and rheology of marmalade were measured. Sensory evaluation of marlamade was compared using a five point scale. Also, the amounts of pectin and naringin in four parts of hyuga-natsu were measured. Changes in texture and structures when soaked, pressurized or boiled were also measured. Firmness of peel was (greatest to least); pressurized > soaked > boiled. The cell walls of flavedo and albedo did not loosen after pressurization. However, after soaking or heating, the middle lamella of albedo separated. The amount of pectin was greatest in albedo > flavedo > endocarp >
juice sacs, respectively. Water-soluble pectin and naringin were found to be slight in all parts. The peel of high-pressure-induced marmalade maintained a natural color. However, a great difference in viscosity between heat-induced and high-pressure-induced marmalade was not found. High-pressure-induced marmalade was evaluated better than heat-induced marmalade by a sensory test.
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Generally, phospholipid, either glycero type or sphingo ones, has diester type of phosphoric acid occupied both sides with alcohol moieties, and constitutes cell membrane as major components. In contrast, phosphatidic acid (PA) and ceramide 1-phosphate (C1P), and their lyso types, lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), all have monoester type of phosphoric acid. Although their amounts in a body are quite minute, all these monoester type of phospholipids are biologically active. Among them, LPA and S1P have been received much attention
because of the simplicity of their chemical structures and the variety of their biological activities.
Zinc coordinated complex, Phos-tag, was synthesized by referring the configuration of zinc metals in the active site of alkaline phosphatase. This reagent can capture phosphoric compounds in specific and dose-dependent manners. Using Phos-tag, we reported a simple method to isolate trace amounts of LPA and S1P by phase partition 1). In extension, we applied immobilized Phos-tag, which was chemically coupled with Sepharose resin, to isolate PA and C1P by solvent elution.
For analyses of phospholipids, LC and TOF mass spectrometric methods coupled with two soft ionizations, ESI and MALDI, are available, and ESI LC MS and MALDI-TOF MS are the general choices of the combination. MALDI-TOF MS is simple to handle and can analyze many samples in a short time. A drawback of MALDI ionization is to use matrix. The advantage to use Phos-tag is that the adduct has high mass range over m/z 1000, where is low in background noise due to matrix. Furthermore, by using monoisotopic zinc, such as 68Zn2+, single molecular ion due to respective molecular species is detected in positive mode with no production of other cation adducts.
Using the phosphate capture molecule, Phos-tag, both at isolation and identification steps, a simultaneous analysis of bioactive lipids having monoester type phosphate is being developed. Details will be presented in the meeting.
1) J. Morishige, T. Tanaka, and K. Satouchi, Methods Mol. Biol. 2012, 874, 45–54.
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Background: Matrix-assisted laser desorption and ionization mass spectrometry coupled with a time-of-flight mass spectrometry (MALDI-TOF MS) has been primarily established for the analysis of compounds with high molecular mass such as protein.
Recently, MALDI-TOF MS have been evaluated as a useful methodology in lipid
research due to its simplicity and high performance. In the present session, we would like to show the MALDI-TOF MS analysis of phosphatidic acid (PA), a potential anti-ulcer phospholipid produced by phospholipase D (PLD) in cruciferous vegetables such as cabbage.
Experimentals: Cabbage juice was obtained by squeezing raw cabbage leaves. Cabbage juice was incubated with egg yolk or soybean lecithin. The products were extracted by Bligh and Dyer procedure and analyzed by MALDI-TOF MS using Phos-tag coordinated with the single isotopic zinc, 68Zn. Phos-tag was coupled chemically to Toyopearl resin and the bead was used for isolation of PA.
Results and Discussion: PA is monoester type of phospholipid and has two negative charges in phosphate, thus, three positive charges are necessary to produce a single positively complex ion. As a result, multiple ions due to additions of protons, sodium and potassium ions, and their combinations are detected in a mass spectrum of PA in positive mode. This is the problem for analysis of molecular species, where PA comprised several molecular species with many fatty acids. Zinc coordinated complex, Phos-tag, was synthesized by referring the spatial configuration of zinc metals in the active site of alkaline phospahatase. This reagent can capture phosphate compound specifically and dose-dependently. Phos-tag can unify adducts of PA to [PA2-/Phos-tag3+]+ in positive mode of MS. Using 68Zn Phos-tag, PA formation by the action of PLD from cabbage juice can follow in a short time by MALDI-TOF MS without purification step. For a trace amount of PA, suppression effects of co-existing other phospholipids are unavoidable.
To overcome, Phos-tag Toyopearl bead is available. An equal amount of mixture of phospholipids, phosphatidylcholine (PC) and PA was incubated with the bead in methanol. PC, diester type of phospholipids, is eluted from the bead thoroughly, whereas PA having monoester type of phosphate interacts with Phos-tag. The retained PA in the bead was recovered by addition of 0.01 M HCl in methanol. Using Phos-tag Toyopearl for isolation, technique for quantitation of PA by MALDI-TOF MS has been under investigations.
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Methods: Subjects were 428 Japanese men and women who participated in a health survey at workplace. Multiple regression was used to examine the association between fatty acid composition in serum (cholesteryl ester, phospholipid, free) and depression (Center for Epidemiologic Studies Depression scale score, 16 or greater) while controlling for potential confounding variables.
Results: High levels of -linolenic acid (in free fatty acids) was associated with fewer depressive symptoms; odds ratio for depression for the highest compared with the lowest tertile of -linolenic acid was 0.5 (trend p = 0.02). EPA and DHA were not associated with lower odds of depression.
Conclusion: -linolenic acid, but not marine-derived, long-chain polyunsaturated fatty acids, may have a protective role against depression in a population with high fish consumption.
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Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are growth-factor like bioactive lipids having phosphate monoester residue. Phos-tag can bind to them and be used both for purification and quantification. In a two-phase solvent system consisting of chloroform/methanol/water, addition of Phos-tag move LPA and S1P from a hydrophilic phase to a hydrophobic phase in the form of their Phos-tag complexes. Using this property, we developed a method for purification of LPA and S1P in biological materials by phase separation technique. Advantages of use of Phos-tag for detection of LPA and S1P in MALDI-TOF MS are an increase in detection efficiency and detection as a single ion form. Homologues of LPA and S1P in natural samples can be quantified by MALDI-TOF MS by using internal standards.
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