Determination of synergistic effects of polymethoxylated flavone extracts of Jinchen orange peels (<Emphasis Type="Italic">Citrus Sinensis</Emphasis> Osberk) with amino acids and organic acids using chemiluminescence

Antioxidant capacity of polymethoxylated flavones (PMFs) extracts of Jinchen orange peels (Citrus sinensis Osberk) was evaluated by using a chemiluminescence (CL) method in vitro. PMFs were isolated using a procedure that produced a consistent mixture of PMFs both in identity and in proportion. The mixture mainly consisted of nobiletin (55.9%), sinensetin (15.2%), heptamethoxyflavone (13.2%), and tangeretin (8.1%). The antioxidative effects of PMFs and the main individual flavones in PMFs as well as synergistic effects of PMFs with organic acids (citric acid, oxalic acid, and malic acid) and amino acids (alanine, lysine, serine, and threonine) on superoxide anion were determined by the pyrogallol–luminol system. The results showed that the inhibition by PMFs on superoxide radical was stronger than that of the individual flavones. Mixtures of citric acid and lysine with PMFs had higher synergistic effects than any other tested mixtures, and the optimum concentrations were exhibited by combinations of 7 μg/mL PMFs + 120 μg/mL citric acid (1.2, p < 0.05) and 7 μg/mL PMFs + 42 μg/mL lysine (1.1, p < 0.05). For the other amino acids and organic acids, the values of the measured inhibition effect (% MIE) of the mixtures with PMFs were not significantly different from their expected inhibition effect (% EIE), which did not show synergism.     

北虫草醇提物体外抗肿瘤和抗病毒活性的比较研究

采用体外人肠腺癌细胞株SW620和人乳腺癌细胞株MCF-7的增殖抑制试验模型和抗病毒(抗流感甲型病毒和抗疱疹病毒)的筛选模型,对北虫草[Cordyceps militaris(L.ex)Link]七个菌株(CM1-CM7)的菌丝体醇提物(CME1-CME7)的抗肿瘤、抗病毒活性进行了比较研究。抗肿瘤试验表明,在中、高浓度时(50μg/mL、200μg/mL,七个醇提物CME1~CME7对SW620和MCF-7的增殖均有一定的抑制活性,其中CME7活性最高(66.0±2.3)%,与阳性对照5-Fu(69.1±4.5)%抑制活性相当。抗病毒试验表明,七个菌株的醇提物中,仅CME7和CME6具有抗流感甲型病毒活性,半数有毒浓度(TC50)分别为288.68μg/mL和>500μg/mL,选择指数(SI)分别为2.23和>3.87;七个菌株的醇提物均未表现出对疱疹病毒HSV-Ⅰ和HSV-Ⅱ的抗性。因此,在七个菌株中,CME7的抗肿瘤活性和抗病毒活性均较好,有必要对其进一步研究研究和开发。     

Effects of eicosapentaenoic acid and docosahexaenoic acid on responses of LPS-stimulated intestinal B lymphocytes from broiler chickens studied in vitro

Effects of different ratios and concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on intestinal B lymphocyte proliferation, IgA secretion and expression of CD5 + CD79a were studied in vitro using cells isolated from broilers. Proliferation was determined by the 3-(4,5- dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay, and those groups with significant difference were selected to detect the IgA using ELISA kit. According to the significant difference of IgA secretion, the expression of CD5 and CD79a was analyzed by fluorescent antibody staining and flow cytometry. The results showed that EPA and DHA inhibited the in vitro proliferation of lipopolysaccharide (LPS)-stimulated B lymphocytes. EPA and DHA significantly suppressed the ability of LPS-stimulated B cells to secrete IgA with ratios of 1:1 at 20 μg/mL and 2:1 at 20 and 40 μg/mL (P < 0.05). In addition, CD5 and CD79a were maximally expressed on LPS-stimulated B cells when the EPA/DHA ratio was 2:1 at 20 μg/mL (P < 0.01), suggesting that the inflammatory reaction may be downregulated by the increasing expression of CD5 and CD79a and inhibitive secretion of IgA by B cell.     

河蚬糖蛋白体外免疫功能研究

目的:研究河蚬糖蛋白对小鼠体外免疫功能的影响。方法:无菌分离小鼠脾淋巴细胞和腹腔巨噬细胞,与不同质量浓度(5、25、125、625μg/mL)的河蚬粗糖蛋白和高纯度糖蛋白CFP共同培养,采用MTT法测定其对淋巴细胞增殖和腹腔巨噬吞噬能力的影响。结果:在无有丝分裂原诱导下,125μg/mL的粗糖蛋白和4个浓度的CFP都有促进脾淋巴细胞的作用。在有丝分裂原刺激下,粗糖蛋白和CFP的刺激指数都比未加有丝分裂原时增大。125μg/mL粗糖蛋白和4个浓度的CFP都有提高巨噬细胞的吞噬能力,且2种河蚬糖蛋白都表现出一定的剂量效应关系。结论:河蚬粗糖蛋白和CFP均有免疫增强活性,其中CFP的活性较强。     

香叶醇联合冬凌草甲素对MCF-7细胞协同抗癌作用及其机理

目的:探讨香叶醇(geraniol,GOH)联合冬凌草甲素(oridonin,ORI)对人乳腺癌MCF-7细胞活力、细胞凋亡、细胞内活性氧(reactive oxygen species,ROS)含量、线粒体膜电位及细胞周期的影响.方法:细胞分为对照组、GOH组(102.81 μg/mL)、ORI组(4.32 μg/m...     

New Phenylethanoid Glycosides and Sugar Esters from Ku-Ding-Cha, a Herbal Tea Produced from Ligustrum purpurascens

ABSTRACT:  Ku-Ding-Cha is a kind of herbal tea used commonly in China for its effects on hypertension, fatness, inflammation, and diuresis. The chlorophyll removal fraction from 60% aqueous acetone extract of Ku-Ding-Cha produced from the leaves of Ligustrum purpurascens (Oleaceae) exhibited observable antioxidant activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay (SC₅₀= 45 μg/mL). Further detailed phytochemical investigation on this fraction led to the isolation of 2 new phenylethanoid glycosides, ligupurpurosides C-D (1 and 2), and 2 new sugar esters, ligupurpurosides E-F (3 and 4), together with 15 known compounds (5 to 19). All of them, except for 7, 8, 9, 11, and 12, were isolated from Ku-Ding-Cha and its original plants for the first time. The chemical structures were elucidated based on the analysis of spectroscopic data, including 1D, 2D NMR, and FABMS. In addition, the isolated compounds were evaluated for their antioxidant activities by DPPH assay.     

响应面试验优化脆江蓠多糖提取工艺及其对肿瘤细胞的抑制作用

目的：研究脆江蓠多糖（Gracilaria chouae polysaccharides,GLP）的提取方法及其对肿瘤细胞生长的影响。方法：响应面法优化GLP提取工艺条件;四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法检测GLP对体外培养的人宫颈癌（HeLa）、食道癌（EC-109）、肝癌（HepG-2）以及乳腺癌（MCF-7）细胞生长的影响。结果：GLP最佳提取条件为预热温度93.1 ℃、料液比1∶61.5（g/mL）、超声时间35 min,在此条件下GLP提取率为16.75%。GLP在质量浓度200～1 000 μg/mL范围内对实验细胞均有抑制作用,且呈剂量依赖关系。当GLP质量浓度为1 000 μg/mL时,其对HeLa、HepG-2、MCF-7和EC-109细胞的抑制率分别为49.11%、41.18%、34.98%和31.33%。对HeLa细胞形态学观察以及Annexin V-FITC/PI荧光染色检测发现,随着GLP给药剂量的增加,凋亡和坏死细胞不断增多。结论：GLP对体外培养的人HeLa、EC-109、HepG-2以及MCF-7等癌细胞增殖有抑制作用,并能诱导肿瘤细胞凋亡。     

Screening of Korean edible wild plants for estrogenic activity using MCF-7 cell proliferation assay

This present study was performed to investigate estrogenic activity of Korean edible wild plants for alternative of estrogen replacement therapy in postmenopausal women. When the estrogenic activity was measured by estrogen induced proliferation of MCF-7 cells, of investigated 29 extracts, 3 samples of kudzu vine (Pueraria thunbergiana), Cardamine leucantha, and David Vetchling (Lathyrus davidii) exhibited a significant proliferation of above 20% at concentration of 100 μg/mL. This stimulation of MCF-7 cell proliferation could be completely blocked by addition of estrogen antagonist ICI 182,780 (100 nM), indicating estrogen receptor-dependent mechanism of these estrogenic effects on MCF-7 cell proliferation.     

INACTIVATION OF BACTERIAL SPORES BY COMBINED ACTION OF HYDROSTATIC PRESSURE AND BACTERIOCINS IN ROAST BEEF

Foodborne bacterial spores are normally resistant to high hydrostatic pressure; however, at moderate pressure, they can be induced to germinate and outgrow. At this stage, they can be killed by bacteriocin-based biopreservatives (BP-containing pediocin and nisin at 3:7 ratio; BP_X, BP + 100 μg/mL lysozyme; BP_Y, BP_X+ 500 μg/mL Na-EDTA). Based on this principle, spores of the meat spoilage organism, Clostridium laramie (1–2 × 10² spores/bag) alone or a mixture of four clostridial spores (5 × 10³ spores/bag), Clostridium sporogenes, Clostridium perfringens, Clostridium tertium, and Clostridium laramie, were inoculated in roast beef in the presence of 5000 AU/g of bacteriocin-based biopreservatives. The roast beef samples were subjected to hydrostatic pressure (HP) at 345 MPa for 5 min at 60C and stored at 4 or 12C for 84 days or at 25C for 7 days. The HP treatment of roast beef samples inoculated with a mixture of clostridial spores could be stored for 42 days at 4C. The HP in combination with either BP_X or BP_Y extended the shelf-life of roast beef up to 7 days at 25C. The combined treatment of HP and BP controlled the growth of C. laramie spores and extended the shelf-life of roast beef for 84 days when stored at 4C.     

Total Phenolic Compounds, Flavonoids, and Radical Scavenging Activity of 21 Selected Tropical Plants

ABSTRACT:  Free radical scavenging activity of 21 tropical plant extracts was evaluated using 1,1-diphenyl-2-picrylhydrazyl assay (DPPH). Total phenolic compounds and flavonoids were determined using Folin–Ciocalteu and HPLC, respectively. Results of the study revealed that all the plants tested exhibited excellent antioxidant activity with IC₅₀ in the range of 21.3 to 89.6 μg/mL. The most potent activity was demonstrated by  Cosmos caudatus  (21.3 μg/mL) and  Piper betle  (23.0 μg/mL) that are not significantly different than that of     -tocopherol or BHA.  L. inermis  extract was found to consist of the highest concentration of phenolics, catechin, epicatechin, and naringenin. High content of quercetin, myricetin, and kaempferol were identified in  Vitex negundo ,  Centella asiatica , and  Sesbania grandiflora  extracts, respectively. Luteolin and apigenin, on the other hand, were found in  Premna cordifolia  and  Kaempferia galanga  extracts. Strong correlation ( R  = 0.8613) between total phenolic compounds and total flavonoids ( R  = 0.8430) and that of antioxidant activity of the extracts were observed. The study revealed that phenolic, in particular flavonoids, may be the main contributors to the antioxidant activity exhibited by the plants.
Practical Application: Potent antioxidant from natural sources is of great interest to replace the use of synthetic antioxidants. In addition, some of the plants have great potential to be used in the development of functional ingredients/foods that are currently in demand for the health benefits associated with their use.     

A comparative study on cytotoxicity and antiproliferative activities of crude extracts and fractions from Iranian wild-growing and cultivated <Emphasis Type="Italic">Agaricus</Emphasis> spp.

Little is known about anticancer capability of wild-growing mushrooms belonging to Agaricus spp. in comparison with their cultivated counterparts. The present study was done to evaluate in vitro antiproliferative effects of dichloromethane-methanol (1:1, v/v) extracts obtained from several Iranian wild strains of Agaricus spp. in comparison with the cultivated white button mushroom, A. bisporus. The wild brown A. bisporus at 500 µg/mL moderately prevented the growth of human prostate cancer (PC3) and human breast cancer (MCF-7) cell lines by 23.54 and 24.57%, respectively, while the wild strains of A. devniensis and A. gennadii at 500 µg/mL had only marginal inhibitory effects. On the contrary, the cultivated A. bisporus significantly inhibited the cell growth at concentrations as low as 125 µg/mL (p?<?0.05). Further, seven distinct fractions were determined based on thin layer chromatography analysis of the cultivated A. bisporus extract. The only active fraction was found to be eluted by petroleum ether (PE):ethyl acetate (EtOAc) (3:2 v/v) which prevented the growth of MCF-7 and PC3 by 74.00 and 63.5%, respectively at 500 µg/mL. Besides, the fraction imposed obvious cytopathic effects in both cell lines at all the tested concentrations, starting from 125 µg/mL. The findings of this study may validate that the known antioxidant potency of wild Agaricus spp. would not necessarily be correlated to their anticancer activity. This research further validated that some semi-polar antiproliferative compounds could be present in the PE/EtOAc-fraction of the cultivated white button mushroom, which warrants further investigations.     

树蝴蝶多糖LKY-I的结构和抗氧化、抗肿瘤活性评价

采用超声波辅助水提、乙醇沉淀、DEAE-52阴离子交换层析法从树蝴蝶(Lobaria koorkauae Yoshim)中分离纯化得到多糖LKY-I。采用凝胶渗透色谱法(gel permeation chromatography,GPC)结合葡聚糖凝胶Sephadex G-100分离,测定其分子质量分布及纯度;利用傅里叶变换红外光谱、气相色谱和核磁共振对其结构进行解析;通过体外清除1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、2’-联氨-双-3-乙基苯并噻唑啉-6-磺酸自由基(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)free radical,ABTS+·)以及对铁离子还原力(ferric-reducing antioxidant power,FRAP)测定结果,评价LKY-I的抗氧化能力;采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT)测定LKY-I对肿瘤细胞Hep G-2、HCC827、MCF-7增殖的抑制作用。结果显示:LKY-I的总糖含量为88.91%,重均分子质量为61 598 D,是组分均一的多糖,由鼠李糖、木糖、甘露糖、葡萄糖、半乳糖组成,其物质的量比为1.1∶2.0∶46.5∶28.5∶20.5。傅里叶变换红外光谱分析结果显示,LKY-I具有一般多糖类物质的特征吸收峰,单糖残基以吡喃环的形式存在。高碘酸氧化、Smith降解以及核磁共振分析表明,LKY-I是含有甘露糖的D-吡喃糖环,包括α和β两种构型,由24.68%的(1→6)或(1→)连接的糖苷键组成。在250~10000μg/m L的质量浓度范围内,随着多糖质量浓度的升高,LKY-I对DPPH自由基、ABTS+·的清除率逐渐增大,当质量浓度为10000μg/m L时,清除率分别达44.12%、24.74%,FRAP值可达0.071 17 mmol/L,与100μg/m L VC(0.072 31 mmol/L)相当。在125~2000μg/m L范围内,随着多糖质量浓度的增加,LKY-I对癌细胞的增殖抑制率逐渐增大。当质量浓度为2000μg/m L时,LKY-I对肿瘤细胞Hep G-2、HCC827、MCF-7增殖的抑制率达到半抑制浓度,说明树蝴蝶水洗多糖LKY-I具有一定的抗肿瘤活性。     

Effects of High-Intensity Pulsed Electric Fields on Lipoxygenase and Hydroperoxide Lyase Activities in Tomato Juice

ABSTRACT:  The influence of high-intensity pulsed electric field (HIPEF) parameters, namely, pulse frequency, pulse width, and polarity on tomato juice lipoxygenase (LOX) and hydroperoxide lyase (HPL) activities was studied using a response surface methodology. Samples were subjected to square-shaped pulses of 35 kV/cm for 1000 μs, with pulse width ranging from 1 to 7 μs at frequencies from 50 to 250 Hz, either in monopolar or bipolar mode. Tomato LOX was more resistant to HIPEF than HPL within the range of assayed conditions. An increase in frequency or pulse width resulted in a decrease of both residual LOX ( RA_LOX ) and HPL ( RA_HPL ) activities. The lowest  RA_LOX  (81%) was observed when tomato juice was treated at 250 Hz for 7 μs in bipolar mode. Moreover, the same conditions led to the highest HPL activity reduction ( RA_HPL  = 10%). A validation of the predictive models determined that 2nd-order expressions were accurate enough to fit the experimental results.     

Comparative Flavonoids Contents of Selected Herbs and Associations of Their Radical Scavenging Activity with Antiproliferative Actions in V79-4 Cells

ABSTRACT:  To elucidate the health benefit of herbal teas on the cytotoxicity induced by H₂O₂ in V79-4 cells, herbal extracts and its flavonoids were tested using lactate dehydrogenase release and determining intracellular reactive oxygen species generation and antioxidant activity with superoxide radical scavenging assay. Significant decrease in cell viability was observed on V79-4 cells treated with H₂O₂ (1 mM), while herbal extracts and its flavonoids including catechin and epigallocatechin gallate prevented the LDH release from H₂O₂ cytotoxicity. Total catechin contents of green tea (65.6 mg/g of dry matter) were significantly higher than other herbal teas (35.8 to 1.2 mg/g of DM). The relative concentration of the 4 major tea catechins ranked EGCG > EGC > EC > C. Green tea exhibited the lowest IC₅₀ values (2 g fresh herb/100 mL) of superoxide radical scavenging activity among the tested herbal tea, which indicates powerful antioxidant activity in O₂^·− radicals scavenging, followed by black tea, dandelion, hawthorn, rose hip, chamomile.     

DECONTAMINATION OF A MULTILAMINATED ASEPTIC FOOD PACKAGING MATERIAL AND STAINLESS STEEL BY OZONE

A multilaminated aseptic food packaging material and stainless steel were treated with ozone to inactivate natural contaminants, bacterial biofilms and dried films of Bacillus subtilis spores and Pseudomonas fluorescens. Sterility of the multilaminated packaging material was achieved when 1.0 x 2.0 cm-pieces of the naturally-contaminated material were treated with ozone in water (5.9 μg/mL) for 1 min. Dried films of spores (10⁸/6.3–cm² surface) were eliminated by 13 μg/mL of ozone in water for the multilaminated packaging material and 8 μg/mL in case of the stainless steel. Ozone inactivated Pseudomonas fluorescens in biofilms more effectively on stainless steel than on the multilaminated packaging material. Repeated exposure to ozone of Pseudomonas fluorescens in biofilms on the multilaminated packaging material eliminated up to ∼10⁸ cfu/12.5 cm². In conclusion, ozone is an effective sanitizer with potential applications in the decontamination of packaging materials and equipment food-contact surfaces.     

Inactivation of Escherichia coli O157:H7 and Salmonella enteritidis in Liquid Egg White Using Pulsed Electric Field

ABSTRACT: The effects of temperature and pulsed electric field (PEF) intensity on inactivation of pathogens such as Escherichia coli O157:H7 and Salmonella enteritidis in egg white was investigated. Liquid egg white inoculated with 10⁸ colony-forming units (CFU)/mL of each pathogen was treated with up to 60 pulses (each of 2 JAS width) at electric field intensities of 20 and 30 kV/cm. The processing temperatures were 10°C, 20°C, and 30°C. After treatment, uninjured and total viable cells were enumerated in selective and nonselective agars, respectively. Maximum inactivations of 3.7 and 2.9 log units were obtained for S. enteritidis and E. coli O157:H7, respectively, while injured cells accounted for 0.5 and 0.9 logs for E. coli O157:H7 and S. enteritidis , respectively. For both bacteria, increasing treatment temperature tended to increase the inactivation rate. There was synergy between electric field intensity and processing temperature. The inactivation rate constant k _T values for E. coli O157:H7 on both selective and nonselective agars were 8.2 × 10^-3 and 6.6 × 10^-3/μS, whereas the values for S. enteritidis were 16.2 × 10^-3 and 12.6 × 10^-3/μS, respectively. The results suggest that E. coli O157:H7 was more resistant to heat-PEF treatment compared with S. enteritidis.     

Differential effects of polyphenols-enriched extracts from hawthorn fruit peels and fleshes on cell cycle and apoptosis in human MCF-7 breast carcinoma cells

This study was to investigate the anticancer effects of the peel polyphenolic extract (HPP) and flesh polyphenolic extract (HFP) from hawthorn fruit in human MCF-7 breast cancer cells. It was found that the polyphenol and flavonoid contents of HPP were significant higher than that of HFP. Both HPP and HFP inhibited cell growth in a dose-dependent manner with the IC₅₀ of 88.6 μg/mL and 175.5 μg/mL, respectively, suggesting that HPP was more effective against MCF-7 cells than HFP. Flow cytometric analysis revealed that both HPP and HFP mediated the cell-cycle arrest at the S-phase, and also dose-dependently led to apoptosis of MCF-7 cells via the mitochondrial pathway, as evidenced by the activation of caspase-3 and caspase-9 and the elevation of intracellular ROS production. All these findings indicate that hawthorn fruit, especially its peel, is an excellent source of natural chemopreventive agents in the treatment of breast cancer.     

Antioxidant and antimicrobial activities of the methanol extracts from pummelo (Citrus grandis Osbeck) fruit albedo tissues

The present study was conducted to evaluate the antioxidant and antimicrobial properties of methanol (100 and 80% aqueous) extracts of pummelo fruits albedo (Citrus grandis Osbeck). The antioxidant and antibacterial activity for crude extracts and isolated compounds were evaluated using free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Paper disc diffusion method. A 100% Methanol (MeOH) extract was steeped in water at different pH levels and partitioned with ethyl acetate (EtOAc) to give basic, acidic, neutral, and phenolic fractions. The neutral extract was found to possess maximum antioxidant and antibacterial activity. Thereafter, neutral extract was carried out on a silica gel column and eluted with hexane:EtOAc:acetone and preparative TLC (PTLC) to give oil buntan compound, linoleic acid methyl ester, β-sitosterol, sigmasterol, limonin, nomilin and meranzin hydrate were isolated. While, 80% MeOH extract was fractionated also using a silica gel column and PTLC to give isomeranzin hydrate, p-coumaric acid and caffeic acid compound. The extract concentration providing 50% inhibition (IC₅₀) was as follows; oil buntan compound 95 μg/mL, caffeic acid 45 μg/mL, p-coumaric acid 105 μg/mL, limonin + nomilin (mixture) 135 μg/mL was lower than that of synthetic antioxidant butylated hydroxyanisole (BHA) 40 μg/mL. The inhibitory zone (mm) of bacteria tested was 2.9–4.1 mm caffeic acid and 11.6–15.1 mm p-coumaric acid.     

多糖中金属离子对其抗氧化活性及抗肿瘤活性的影响

采用水提醇沉法制备香菇、海带、枸杞多糖粗品,Sevage法除蛋白制得初级纯化多糖。利用EDTA-2Na去除多糖粗品和纯品中的金属离子,比较去除前、后多糖抗氧化活性及抗肿瘤活性的变化。结果表明:3种多糖均含有Al、Ca、Mn等多种金属元素,只是含量各不相同。去除金属离子后,多糖对ABTS自由基、DPPH自由基的清除能力均下降,对羟自由基的清除能力变化不一。其中枸杞多糖质量浓度为600μg/mL时抗氧化活性变化最为显著,ABTS自由基清除率由26.31%降至17.25%,DPPH自由基清除率由59.80%降至38.40%。枸杞多糖纯品在去除金属后对人乳腺癌细胞MCF-7的体外抑制活性呈下降趋势。结论:多糖中的金属元素是多糖活性中心构成的一部分,对其活性具有重要影响。