Protocatechuic acid, a metabolite of anthocyanins, inhibits monocyte adhesion and reduces atherosclerosis in apolipoprotein E-deficient mice

Polyphenols, including anthocyanins, from various plant foods are effective in the prevention of atherosclerosis in animal and human studies. Protocatechuic acid (PCA), a major metabolite of anthocyanins, has been found to possess the anti-carcinogenic effect, whereas the in vivo effect of PCA as an anti-atherosclerotic agent remains unknown. We demonstrated herein that PCA inhibited monocyte adhesion to tumor necrosis factor-α (TNF-α)-activated mouse aortic endothelial cells, associated with the inhibition of vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) expression. Furthermore, PCA inhibited the nuclear content of p65, a subunit of nuclear factor-κB (NF-κB), along with reduced NF-κB binding activity. Finally, PCA administration in the apolipoprotein E (ApoE)-deficient mouse model reduced aortic VCAM-1 and ICAM-1 expression, NF-κB activity, and plasma-soluble VCAM-1 and ICAM-1 levels, with inhibiting atherosclerosis development. We suggest that PCA possesses the anti-atherogenic effect at least partially via its anti-inflammatory activity.     

Serotonin derivatives, major safflower (Carthamus tinctorius L.) seed antioxidants, inhibit low-density lipoprotein (LDL) oxidation and atherosclerosis in apolipoprotein E-deficient mice

The effects of defatted safflower seed extract and its phenolic constituents, serotonin derivatives, on atherosclerosis were studied. Ethanol-ethyl acetate extract of safflower seeds (SSE) inhibited low-density lipoprotein (LDL) oxidation induced in vitro by an azo-containing free-radical initiator V70 or copper ions. Two serotonin derivatives [N-(p-coumaroyl)serotonin, CS; N-feruloylserotonin, FS] and their glucosides were identified as the major phenolic constituents of the extract. The study with chemically synthesized materials revealed that a majority of the antioxidative activity of SSE was attributable to the aglycones of these two serotonin derivatives. Orally administered CS and FS suppressed CuSO(4)-induced plasma oxidation ex vivo. Long-term (15 week) dietary supplementation of SSE (1.0 wt %/wt) and synthetic serotonin derivatives (0.2-0.4%) significantly reduced the atherosclerotic lesion area in the aortic sinus of apolipoprotein E-deficient mice (29.2-79.7% reduction). The plasma level of both lipid peroxides and anti-oxidized LDL autoantibody titers decreased concomitantly with the reduction of lesion formation. Serotonin derivatives were detected as both intact and conjugated metabolites in the plasma of C57BL/6J mice fed on 1.0% SSE diet. These findings demonstrate that serotonin derivatives of SSE are absorbed into circulation and attenuate atherosclerotic lesion development possibly because of the inhibition of oxidized LDL formation through their strong antioxidative activity.     

Cacao polyphenols influence the regulation of apolipoprotein in HepG2 and Caco2 cells

Cocoa powder is rich in polyphenols, such as catechins and procyanidins, and has been shown to inhibit low-density lipoprotein (LDL) oxidation and atherogenesis in a variety of models. Human studies have also shown daily intake of cocoa increases plasma high-density lipoprotein (HDL) and decreases LDL levels. However, the mechanisms responsible for these effects of cocoa on cholesterol metabolism have yet to be fully elucidated. The present study investigated the effects of cacao polyphenols on the production of apolipoproteins A1 and B in human hepatoma HepG2 and intestinal Caco2 cell lines. The cultured HepG2 cells or Caco2 cells were incubated for 24 h in the presence of cacao polyphenols such as (-)-epicatechin, (+)-catechin, procyanidin B2, procyanidin C1, and cinnamtannin A2. The concentration of apolipoproteins in the cell culture media was quantified using an enzyme-linked immunoassay, and the mRNA expression was quantified by RT-PCR. Cacao polyphenols increased apolipoprotein A1 protein levels and mRNA expression, even though apolipoprotein B protein and the mRNA expression were slightly decreased in both HepG2 cells and Caco2 cells. In addition, cacao polyphenols increased sterol regulatory element binding proteins (SREBPs) and activated LDL receptors in HepG2 cells. These results suggest that cacao polyphenols may increase the production of mature form SREBPs and LDL receptor activity, thereby increasing ApoA1 and decreasing ApoB levels. These results elucidate a novel mechanism by which HDL cholesterol levels become elevated with daily cocoa intake.     

Apple peel polyphenols protect against gastrointestinal mucosa alterations induced by indomethacin in rats

The stability of an apple peel polyphenol extract (APPE) with powerful antioxidant activity was evaluated under acidic conditions in vitro, and its protective effect against gastrointestinal damage was investigated in rats treated with indomethacin. The antioxidant activity of APPE remained stable at pH 2.0 for 4 h. In rats treated with indomethacin (40 mg/kg ig), the previous administration of APPE protected the gastric, intestinal, and colonic mucosa from oxidative stress by preventing increased malondialdehyde concentrations and decreasing the GSH/GSSG ratio. APPE also displayed anti-inflammatory effects by preventing neutrophil infiltration in the mucosa, as evidenced by the lower myeloperoxidase activity. These protective effects of APPE resulted in the prevention of macro- and microscopic damage and of barrier dysfunction along the gastrointestinal tract of the indomethacin-treated animals. This study supports the concept that apple peel polyphenols may be useful in the prevention and/or treatment of nonsteroidal anti-inflammatory drug-associated side effects.     

Apple polyphenols affect protein kinase C activity and the onset of apoptosis in human colon carcinoma cells

Polyphenol-rich apple extracts have been reported to suppress human colon cancer cell growth in vitro. The protein kinase C (PKC) is among the signaling elements known to play an important role in colon carcinogenesis. In the present study, we investigated whether apple polyphenols affect PKC activity and induce apoptosis in the human colon carcinoma cell line HT29. A polyphenol-rich apple juice extract (AE02) was shown to inhibit cytosolic PKC activity in a cell-free system. In contrast, incubation of HT29 cells for 1 or 3 h with AE02 up to 2 mg/mL did not affect the cytosolic PKC activity. After prolonged incubation (24 h), cytosolic PKC activity was modulated, albeit a u-shaped curve of effectiveness was observed, with an initial inhibitory effect followed by the recurrence and even induction of enzyme activity. Concomitantly, in the cytosol, a significant decrease of the protein levels of PKCalpha, PKCbetaII, and PKCgamma together with a significant increase of a proapoptotic PKCdelta fragment was observed. However, the effects on the protein levels of these PKC isoforms in the cytosol were not associated with translocation between the different cellular compartments but might instead result from the onset of apoptosis. Indeed, the treatment with AE02 was shown to induce apoptosis by the activation of caspase-3, DNA fragmentation, and cleavage of poly(ADP ribose) polymerase. So far, identified and available constituents of the apple extract did not contribute substantially to the observed effects on PKC and apoptosis induction. In summary, apple polyphenols were found to inhibit PKC activity in a cell-free system. However, our results indicate that within intact cells PKC does not represent the primary target of apple polyphenols but appears to be affected in the course of apoptosis induction.     

Antiatherogenic effect of isoflavones in ovariectomized apolipoprotein e-deficient mice

The consumption of isoflavone-containing foods such as soybean and soybean products has been reported to have beneficial effects on the cardiovascular system in postmenopausal women. The present study was carried out to examine the mechanism underlying the beneficial effects of isoflavones in apolipoprotein (apo) E-deficient mice subjected to ovarian resection. Compared with sham-operated mice, ovariectomized mice had a larger arterial lesion area in the aortic root. Feeding the ovariectomized mice an isoflavone-containing diet (0.055 mg/kJ of total isoflavones/cal of diet) reduced the size of these lesions more than did feeding them with an isoflavone-free diet. Neither ovariectomy nor diet had a significant effect on the concentration of cholesterol in serum and urinary levels of isoprostanes, which are biomarkers for oxidative stress in vivo. The ovariectomized mice showed a greater increase in mRNA abundance for monocyte chemoattractant protein (MCP)-I in the aorta and in the level of nitric oxide (NO) secreted by peritoneal macrophages in culture than did the sham-operated mice. The isoflavone-containing diet lowered the MCP-I expression and the NO secretion more than did the isoflavone-free diet. These results suggest that dietary isoflavones confer an antiatherogenic effect by preventing the activation of macrophages due to the removal of ovaries.     

Anthocyanins from purple sweet potato Ipomoea batatas cultivar Ayamurasaki suppress the development of atherosclerotic lesions and both enhancements of oxidative stress and soluble vascular cell adhesion molecule-1 in apolipoprotein E-deficient mice

We evaluated the protective potential of anthocyanins from purple sweet potato Ipomoea batatas cultivar Ayamurasaki (APSP) against low-density lipoprotein (LDL) oxidation in vitro and atherosclerotic lesion development in apolipoprotein E-deficient mice given a cholesterol- and fat-enriched diet with or without 1% APSP for 4 weeks. APSP protected LDL against oxidation more potently than other anthocyanins and l-ascorbic acid in vitro. In mice, APSP significantly lowered the atherosclerotic plaque area to about half of the control, the liver level of thiobarbituric acid-reactive substances as an oxidative stress marker, and the plasma level of soluble vascular cell adhesion molecule-1 (sVCAM-1). However, APSP showed no effects on body weight and cholesterol and lipid levels in the plasma. The results suggest that APSP can suppress the development of atherosclerotic lesions and both enhancements of oxidative stress and sVCAM-1 independently of the changes in cholesterol and lipid levels in mice.     

Consumption of a novel dietary formula of plant sterol esters of canola oil fatty acids, in a canola oil matrix containing 1,3-diacylglycerol, reduces oxidative stress in atherosclerotic apolipoprotein E-deficient mice

The antiatherogenic properties of a novel dietary formula (PS-CO) of plant sterol esters of fatty acids, produced by enzymatic interesterification of plant sterols with canola oil (CO), in a CO matrix containing 1,3-diacylglycerol, were evaluated in apolipoprotein E-deficient mice. PS-CO consumption strongly tended to lower total plasma cholesterol levels by 21%, compared to the placebo group. Blood triglycerides were reduced by 38% and 36% compared to CO and placebo-fed mice, respectively. Serum lipid peroxide levels were lowered following PS-CO administration by 62% and 63%, compared to CO and placebo administration, respectively. Unlike CO supplementation, PS-CO consumption preserved serum paraoxonase (PON1) activity. Mouse peritoneal macrophages from PS-CO-fed mice exhibited reduced cellular uptake of oxidized-LDL compared to those from placebo-fed mice and demonstrated a tendency toward a decreased capability to release superoxide anions. These findings indicate that PS-CO supplementation is beneficial in reducing serum lipid levels, and serum and macrophage oxidative stress, thus contributing to the reduction in atherogenic risk factors.     

Pomegranate byproduct administration to apolipoprotein e-deficient mice attenuates atherosclerosis development as a result of decreased macrophage oxidative stress and reduced cellular uptake of oxidized low-density lipoprotein

The effects of a pomegranate byproduct (PBP, which includes the whole pomegranate fruit left after juice preparation) on atherosclerosis development in apolipoprotein E-deficient (E degrees ) mice were studied. Consumption of PBP (17 or 51.5 microg of gallic acid equiv/kg/day) by the mice resulted in a significant reduction in atherosclerotic lesion size by up to 57%. PBP consumption significantly reduced oxidative stress in the mice peritoneal macrophages (MPM): Cellular lipid peroxide content decreased by up to 42%, the reduced glutathione levels increased by up to 53%, and paraoxonase 2 lactonase activity increased by up to 50%, as compared to MPM from E degrees mice that consumed only water. Furthermore, oxidized low-density lipoprotein (Ox-LDL) uptake by the MPM was reduced by up to 19%. Similar results were observed also in vitro. Treatment of J774A.1 macrophages with PBP (10 or 50 micromol/L of total polyphenols) significantly decreased both cellular total peroxide content and Ox-LDL uptake. It was thus concluded that PBP significantly attenuates atherosclerosis development by its antioxidant properties.     

Protective effect of apple polyphenols against stress-provoked influenza viral infection in restraint mice

This study was conducted to investigate the effects of apple polyphenol extract (APE) against influenza virus in mice loaded with restraint stress. The high-performance liquid chromatography (HPLC) fingerprint of APE was recorded, and the percentage composition of polyphenols was determined as 81.7%. Our results showed that restraint stress significantly promoted the mortality and duration of complications of mice infected with the H1N1 virus. However, oral administration of APE (100 and 200 mg/kg) improved the survival rates and prolonged living time of stressed mice infected with influenza virus in a dose-dependent manner. APE was further found to significantly improve the number of immunocytes, ratio of CD4 helper cells, secretion of IL-2, and capabilities of natural killer (NK) cytotoxicity (LU10/spleen) in spleens of restraint-stressed mice. In addition, APE also significantly decreased the level of lipid peroxidation and increased oxygen radical absorbance capacity (ORAC) in splenocytes. These results indicated that the protective effects of APE on mice infected with influenza virus were related to the alleviation of stress-induced impairment of immune functions and its antioxidant property might contribute to the immune recovery.     

Antioxidant polyphenols in almond and its coproducts

Antioxidant efficacy of defatted almond whole seed, brown skin, and green shell cover extracts was evaluated by monitoring inhibition of human low-density lipoprotein (LDL) oxidation, inhibition of DNA scission, and metal ion chelation activities. The total phenolic contents of ethanolic extracts of brown skin and green shell cover of almond were 10 and 9 times higher than that of the whole seed, respectively. Brown skin extract at 50 ppm effectively inhibited copper-induced oxidation of human LDL cholesterol compared to whole seed and green shell cover extracts, which reached the same level of efficacy at 200 ppm. Green shell cover extract at 50 ppm level completely arrested peroxyl radical-induced DNA scission, whereas 100 ppm of brown skin and whole seed extracts was required for similar efficiencies. All three almond extracts exhibited excellent metal ion chelation efficacies. High-performance liquid chromatographic (HPLC) analysis revealed the presence of quercetin, isorhamnetin, quercitrin, kaempferol 3-O-rutinoside, isorhamnetin 3-O-glucoside, and morin as the major flavonoids in all extracts.     

Apple polyphenols and products formed in the gut differently inhibit survival of human cell lines derived from colon adenoma (LT97) and carcinoma (HT29)

Colorectal tumor risks could be reduced by polyphenol-rich diets that inhibit cell growth. Here, apple polyphenols were studied for effects on the survival of colon adenoma (LT97) and carcinoma-derived (HT29) cell lines. Three apple extracts (AEs) from harvest years 2002-2004 were isolated (AE02, AE03, and AE04) and fermented in vitro with human fecal flora. Extracts and fermentation products were analyzed for polyphenols with HPLC. The cells were treated with AEs (0-850 microg/mL) or fermented AEs (F-AEs, 0-9%), and survival was measured by DNA staining. All AEs contained high amounts of polyphenols (311-534 mg/g) and reduced cell survival (in LT97 > HT29). AE03 was most potent, possibly because it contained more quercetin compounds. Fermentation of AEs resulted in an increase of short chain fatty acids, and polyphenols were degraded. The F-AEs were approximately 3-fold less bioactive than the corresponding AEs, pointing to a loss of chemoprotective properties through fermentation.     

新疆苹果皱果类病毒(AFCVd)的检测与序列分析

采集新疆焉耆地区的老果园中的苹果（Malus domestica Brkn）树枝条、叶片和韧皮部,提取总RNA,根据GenBank中的AB104558序列设计引物,利用RT-PCR技术扩增出苹果皱果类病毒（Apple fruit crinkle viroid ,AFCVd）特异条带,通过回收、克隆和测序,结果表明,获得的10条序列,已在GenBank中登录（登录号：EU031507～EU031516）。利用带有AFCVd目的DNA的质粒为模板,成功地合成了地高辛标记的cDNA探针,该探针能很好地用于AFCVd的检测。并利用原位RT-PCR技术做进一步检测,证明苹果叶片中有苹果皱果类病毒存在,主要分布在叶肉细胞的细胞核中。     

苹果锈果类病毒新疆分离物的核苷酸序列分析

苹果锈果类病毒(Apple scar skin viroid,ASSVd)属于Pospiviroidae科(Hashimoto et al.,1987),主要侵染苹果和梨(陈炜等,1988a,b),是我国苹果产区的一种重要病害。刘福昌等(1985)和陈炜等(1988a、b)对ASSVd进行了研究。     

绿肥间作和滴灌对苹果园土壤肥力和果实品质的影响

  目的  研究绿肥间作和水肥一体化技术对苹果园土壤肥力和果树生产的影响。   方法  于2018 ~ 2020年设常规施肥 + 自然生草（CF）、滴灌 + 常规施肥 + 自然生草（SF）、常规施肥 + 绿肥（GF）、滴灌 + 常规施肥 + 绿肥（SGF）和滴灌 + 追肥氮磷钾用量较常规施肥减少20% + 绿肥（80%SGF）5个处理,在苹果园进行小区水肥试验,研究毛叶苕子与果树间作、滴灌施肥对果园土壤肥力及苹果果实品质的影响。  结果  两年的研究结果表明:所有处理中,以SGF处理土壤的微生物数量最多,土壤养分含量最高,苹果的果实品质最好。与CF处理相比,SF处理土壤碱解氮和速效钾含量平均增加了8.46%和10.06%,苹果的可溶性固形物和VC含量平均增加了11.01%和4.37%,土壤放线菌数量、脲酶和磷酸酶活性显著增加,GF处理土壤碱解氮含量平均增加了8.09%,苹果的可溶性固形物和VC含量平均增加了14.86%和9.25%,土壤的微生物数量和土壤的脲酶、磷酸酶活性显著增加;与SF处理相比,SGF处理土壤有机质、碱解氮、有效磷含量显著增加,苹果可溶性固形物和VC含量平均增加了7.74%和8.80%,可滴定酸含量降低了11.12%;与GF处理相比,SGF处理土壤碱解氮、有效磷和速效钾含量显著增加,可溶性固形物和VC含量显著增加。  结论  绿肥间作和水肥一体化技术应用于果树生产,能够显著提高土壤肥力、改善果实品质,其中以滴灌 + 常规施肥 + 绿肥间作处理效果最好。     

Row Distance and Tree Heights in Apple Trees

Abstract

The apple cultivars ‘Discovery’, ‘Jonagold’ and ‘Mutsu’ were planted in 1980 at row distances of 3 and 4.5 m. Tree heights of 1.5, 2 and 2.5 m were established at 3 m row distance, and 2.25, 2.75 and 3.25 m at 4.5 m row distance. The tree heights were effective from 1985–86; yield and fruit development were followed during 1982–90. Yield per tree, tree size and fruitfulness were larger at the wider row distance, especially for ‘Mutsu’. Thus, in ‘Mutsu’, yield per hectare at 4.5 m equalled that at 3 m, contrary to ‘Discovery’ and ‘Jonagold’, where yield per hectare was higher at 3 m. Also, dry matter contents and colour development (‘Jonagold’) were improved by 4.5 m row distance in several cases. Reduced tree heights decreased yields and colour development (‘Jonagold’), but with minor reductions at tree heights of 2.0–2.5 m during the first 7–8 years. A tree height of 1.5 m, at 3 m row distance, responded negatively, especially for ‘Discovery’ and ‘Jonagold’.     

Rapid determination of polyphenols and vitamin C in plant-derived products

Polyphenols, widely spread in our diet by the consumption of plant food products, are commonly determined using Folin-Ciocalteu reagent that interacts with other different reducing nonphenolic substances and leads to an overestimation of polyphenol content. In this paper we report an optimized Folin-Ciocalteu method to specifically determine the contents of total polyphenols and vitamin C. After the optimal conditions for the colorimetric assay were set, solid-phase extraction (Oasis HLB (hydrophilic-lipophilic balance)) was carried out to eliminate the water-soluble reducing interferences including vitamin C. Colorimetric correction was thus performed by subtracting interfering substances contained in the water washing extract from the raw extract. Moreover, vitamin C present in the water washing extract can be destroyed by heating and thus colorimetrically deduced. This procedure was set up with synthetic solutions and validated on different extracts from fruit products.     

纤维红外吸收特性及其在皮棉杂质检测中的应用

皮棉异性纤维杂质检测技术是近几年来国内外研究的难点。为有效检测皮棉中与棉纤维外观极其相似的异性纤维杂质，提出了一种显微近红外成像方法用于检测皮棉中异性纤维。该方法将棉纤维与异性纤维在特定红外波段的吸收特性差别，转化为近红外光谱成像系统中两者的灰度、形态图像特征差别，通过显微光路对图像特征差别放大，利用图像分割技术将异性纤维目标分割出来。试验结果表明，采用显微近红外成像方法捕获的图像中，异性纤维灰度、形态特征明显，其检测结果与实际相符，此方法可有效识别皮棉中异性纤维杂质。     

Effect of red grapes co-winemaking in polyphenols and color of wines

The red grapes co-winemaking effect on phenolic fraction and wine color has been studied for the first time, where Monastrell was comacerated and cofermentated with Cabernet Sauvignon and Merlot. Changes in the relative abundance of anthocyanins were observed as well as hyperchromic shifts at 530 and 620 nm; these effects remain constant after aging. Co-winemaking also favored copigmentation, giving way to more stable anthocyanins and facilitating their polymerization. With regard to color evolution, the mixture of Monastrell with Merlot grapes was more appropriate than with Cabernet Sauvignon for aging wines in oak barrels. The extent of copigmentation was more important in young wines than in aged wines. This is mainly due to the self-anthocyanin monomer reactions in the case of young wines, whereas in aged wines copigmentation is mainly due to the reaction between the anthocyanins and other polyphenolic cofactors. Discriminant analysis showed the possibility of differentiating wines according to the aging time and the type of wine, with color parameters (color intensity, OD 620 nm, and OD 520 nm) being the most important discrimination variables in the first case and petunidin-3-glucoside and peonidin-3-glucoside contents in the second case.     

Principal component analysis of biogenic amines and polyphenols in Hungarian wines

Biogenic amines, polyphenols, and resveratrol were analyzed quantitatively in 25 different Hungarian wines from the same wine-making region, harvest of 1998. Polyphenols were determined according to a spectrophotometric method, whereas other substrates were analyzed using overpressured-layer chromatography (OPLC). Principal component analyses (PCA) were performed on data matrices consisting of substrates (columns) and different sorts of wines (rows) from the region of Pécs (southern Hungary). It was found that four (unrotated) principal components account for >80% of the total variance in the data. The plots of component loadings showed significant groupings for concentrations of biogenic amines (and polyphenols). Similarly, the component scores grouped according to the different sorts of wines. The loading plots reveal that there is no need to measure all of the variables to achieve the same characterization. It is enough to measure one variable per group. Naturally, this conclusion is valid only within the limits of the present study; wines from other regions may behave differently.