The Anti-allergic Activity of Polyphenol Extracted from Five Marine Algae

Natural polyphenol has been widely believed to be effective in allergy remission. Currently, most of the natural polyphenol products come from terrestrial sources such as tea, grape seeds among others, and few polyphenols have been developed from algae for their anti-allergic activity. The aim of the study was to screen some commercial seaweed for natural extracts with anti-allergic activity. Five algae including Laminaria japonica, Porphyra sp., Spirulina platensis, Chlorella pyrenoidosa and Scytosiphon sp. were extracted with ethanol, and the extracts were evaluated for total polyphenol contents and anti-allergic activity with the hyaluronidase inhibition assay. Results showed that the total polyphenol contents in the ethanol extracts ranged from 1.67% to 8.47%, while the highest was found in the extract from Scytosiphon sp. Hyaluronidase inhibition assay showed that the extracts from Scytosiphon sp. had the lowest IC50, 0.67 mg m L-1, while Chlorella pyrenoidosa extract had the highest IC50, 15.07 mg m L-1. The anti-allergic activity of Scytosiphon sp. extract was even higher than the typical anti-allergic drug Disodium Cromoglycate(DSCG)(IC50 = 1.13 mg m L-1), and was similar with natural polyphenol from Epigallocatechin gallate(EGCG)(IC50 = 0.56 mg m L-1). These results indicated that the ethanol extract of Scytosiphon sp. contains a high concentration of polyphenol with high anti-allergic activity. Potentially Scytosiphon sp. can be developed to a natural anti-allergic compound for allergy remission.     

Influence of Ultrasonic Treatment on the Allergenic Properties of Shrimp （Penaeus vannamei ） Allergen

The present study was undertaken to determine whether high intensity ultrasound could reduce the allergic properties of shrimp allergens. Reducing the allergenic properties of these allergens will be beneficial to allergic individuals. Samples of shrimp protein extract and shrimp muscle were treated by high-intensity ultrasound with water bathing at 0 ℃ or 50 ℃ for different time periods. The treated and untreated samples were then analyzed by SDS-PAGE, Western blots and competitive inhibition ELISA （Ci-ELISA） to determine the shrimp allergenicity. The results show that high-intensity ultrasound has no effect on allergenicity when the extracts were treated at 0℃. However, a significant decrease was observed in the level of the major shrimp allergen, Pen a 1, when the samples were treated at 50 ℃. In the determination of allergenicity with CiELISA, a reduction in IgE binding was also observed.     

The effect of simulated gastrointestinal digestion on shrimp <Emphasis Type="Italic">Penaeus vannamei</Emphasis> allergenicity

Pen a 1 is a major shrimp allergen that induces an allergy. The aim of this study is to assess the effect of simulated gastrointestinal fluids on the allergenicity of Pen a 1. Purified Pen a 1 from shrimp (Penaeus vannamei) was subjected to digestion in SGF (simulated gastric fluid) and SIF (simulated intestinal fluid) for a set time. The allergenicity of digestive Pen a 1 was analyzed by immunoblotting and Ci-ELISA, using pool sera from patients with shrimp specific IgE. The results showed that Pen a 1 exhibited a decrease in allergenicity with increasing digestion time in the SGF and SIF. However, Pen a l exhibited strong resistance to digestive fluids, and all yielded fragments (33 kD, 23 kD, and 14 kD) showed allergic activity. Therefore, anti-digestion may be an important factor for Pen a 1 to induce an allergy.     

Research on the critical conditions for clay particle release during saline aquifer freshening process

Water sensitivity phenomenon occurs during saline aquifer freshening process in seawater intrusion area, and clay particles released in the phenomenon can damage the infiltration capacity of the aquifer. In order to find out the factors and mechanisms for clay particle release, laboratory column infiltration experiments simulating saline aquifer freshening process were designed to measure the critical conditions(critical flow velocity, critical salt concentration and critical ionic strength) and force analysis for clay particle according to DLVO electric double layer theory was employed to illustrate the mechanisms for particle release. The research results showed that critical flow velocity for clay particle release is influenced by salt concentration of injecting solution. When salt concentration of injecting solution is very high, clay particles are not released, indicating that there does not exist a critical flow velocity in this situation. As salt concentration of injecting solution decreases, particles start to be released. The critical salt concentration for clay particle release is 0.052 mol L-1 in our work, which was determined by a constant-flux experiment for stepwise displacement of high concentration Na Cl solution. The critical ionic strength for clay particle release decreases as Ca2+ molar content percentage of the mixed solution of Na Cl and Ca Cl2 increases following the first-order exponential decay equation y = 0.0391e-0.266 x + 0.0015.     

微酸性电解水结合复合保鲜剂对凡纳滨对虾冷藏期间蛋白特性影响

【目的】研究微酸性电解水(SAEW)结合迷迭香提取物(RE)-柠檬酸(CA)复合保鲜剂处理对凡纳滨对虾(Litopenaeus vannamei)冷藏期间蛋白特性变化影响,探讨此法用于虾类贮藏保鲜的效果。【方法】将样品分别经SAEW、质量分数4%RE+质量分数1%CA(RC)、SAEW结合RC(SAEW+RC)和无菌水(CK)处理10 min后,自然沥干15 min,装入PE保鲜袋于(4±1)℃冰箱中保藏,每2 d测定样品的pH值、总挥发性盐基氮(TVB-N)、总巯基含量、Ca²⁺-ATPase活性、化学键、肌原纤维小片化指数(MFI)与内源荧光强度(IFI)等指标,并结合聚丙烯酰胺凝胶电泳(SDS-PAGE)进行综合分析。【结果】复合处理能延缓凡纳滨对虾贮藏期间pH与TVB-N值的升高,抑制总巯基含量下降,CK、RC、SAEW与SAEW+RC组样品的Ca²⁺-ATPase活力分别由初始0.16、0.17、0.17与0.17μmol/(mg·min)降至0.12、0.13、0.12与0.14μmol/(mg·min),MFI值较初始值上升1.88、1.73、1.76和1.54倍,可见SAEW+RC对延缓Ca²⁺-ATPase活性下降与MFI值的上升有较好作用。同时,SAEW+RC处理还能减缓蛋白质分解、氧化和变性,保持蛋白质化学键与IFI值的相对稳定。【结论】微酸性电解水结合复合保鲜剂处理能较好抑制凡纳滨对虾冷藏期间的蛋白质氧化分解,延缓其品质劣变。     

一株促藻生长细菌培养条件的优化

研究分离自对虾养殖池的一株促藻生长细菌的最佳培养条件,通过单因子实验得到该菌株的最佳碳源为葡萄糖,最佳氮源为酵母膏,初始pH值为7.5,最佳氯化钠质量浓度5 g/L;在其他培养条件不变的情况下,通过对酵母膏、葡萄糖、初始pH值做三因子三水平的正交实验,得到培养基最佳组合为:酵母膏10 g/L,葡萄糖1 g/L,初始pH值7.5。     

Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)

The clotting protein (CP) plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis (named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca²⁺, suggesting that the clotting reaction is catalyzed by a Ca²⁺-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers. The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.     

Identification of the Major Allergen in Greasy-Back Shrimp （Metapenaeus ensis） by MALDI-TOF-MS

In order to better understand shrimp allergen,some basic characters of the major allergen of greasy-back shrimp (Metapenaeus ensis)were investigated.The major allergen was extracted and separated,and its peptide mass fingerprint(PMF) was analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).After screening in the NCBI database with Mascot searching engine,the results indicated that the major allergen of greasy-back shirmp was muscle tropomyosin.Database matching search showed that the top protein matched,i.e.the tropomyosin from giant tiger prawn(Penaeus monodon),had a Mowse value of 268.In addition,there were 27 queries matched with the allergen in greasy-back shirmp with an amino acid sequence coverage value of 65%.The matching scores and the sequence coverage values were also high with tropomyosins of other invertebrates,including Tyrophagus putrescentiae and Lepisma saccharina.These results indicated that the allergen of Metapenaeus ensis had high homology with other crustacean allergens,and provided molecular explanations for the high cross-reactivity of the major allergens between crustaceans and some other invertebrates.     

Antibacterial activity of extracts from <Emphasis Type="Italic">Zostera marina</Emphasis> against pathogens of <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> skin ulceration disease

The purpose of this study was to investigate the antibacterial activity of extracts from Zostera marina against the pathogens of Apostichopus japonicus skin ulceration disease. When 95% ethanol (v/v) solvent was used to extract Zostera marina at 50°C, aqueous extract (ZA) showed obvious bacteriostatic effects on the tested bacterial strains (inhibition halo diameters between 8.23 mm and 13.62 mm), whereas the ethyl acetate extract (ZE) was almost inactive. The minimal inhibitory concentration (MIC) of ZA against four pathogens were homogeneous at 12.8 g L⁻¹. ZA components were analyzed by thin layer chromatography (TLC) assay and six fractions were obtained. In another study, the six fractions showed inhibitory effects against the tested bacteria while their functions seemed to counteract the ZA activity.     

温度、光照对小环藻生长和叶绿素a含量的影响

用单因子和正交实验方法研究了温度和光照对小环藻生长和叶绿素a含量的影响。结果表明,温度对小环藻生长和叶绿素a含量有极显著影响(P<0.01),当温度在30~40℃时比增长率和叶绿素a含量分别为0.37~0.38 d-1和1 084.4~1 214.2μg.L-1,均显著高于其他温度组。光照对小环藻生长和叶绿素a含量有极显著影响(P<0.01),当光照强度从29.25~146.26μmol.m-2.s-1时比增长率最高,为0.30~0.32 d-1;而叶绿素a含量则在9.75~58.50μmol.m-2.s-1时最高,达709.7~731.8μg.L-1,极显著高于其他照度组。正交实验结果表明:在35℃、87.75μmol.m-2.s-1条件下种群增长最快,比增长率为0.4402 d-1;在30℃、29.25μmol.m-2.s-1时,叶绿素a含量最高,为1 746.6μg.L-1。     

Optimization of the microwave-assisted extraction of phlorotannins from Saccharina japonica Aresch and evaluation of the inhibitory effects of phlorotannin-containing extracts on HepG2 cancer cells

The use of a microwave-assisted extraction(MAE) method for the extraction of phlorotannins from Saccharina japonica Aresch(S.japonica) has been evaluated with particular emphasis on the influential parameters,including the ethanol concentration,solid/liquid ratio,extraction time,extraction temperature,and microwave power.The MAE procedure was optimized using single-factor design and orthogonal array design(OAD).The content of total phlorotannins in S.japonica was determined using a Folin-Ciocalteu(FC) assay.A maximum total phlorotannin content of 0.644 mg of phloroglucinol equivalent per gram of dry weight plant(mg PGE/g DW) was obtained using the optimized model,which included an ethanol concentration of 55%,solid/liquid ratio of 1:8,extraction time of 25 min,irradiation power of 400 W,and temperature of 60°C.Under similar conditions,the application of a conventional extraction method led to a lower phlorotannin yield of 0.585 mg PGE/g WD.These results demonstrated that the MAE approach provided better results for the extraction of phlorotannins from S.japonica and was a promising technique for the extraction of phenolic compounds from S.japonica and other materials.In addition,screening tests for the inhibitory activity showed that the phlorotannin-containing extracts significantly inhibited the growth of human hepatocellular carcinoma cells(HepG2) by inducing their apoptosis.The morphological changes that occurred during cell apoptosis were characterized using Hoechst33258 staining.     

水产饲料中三聚氰胺检测方法的优化

通过对高效液相色谱法(HPLC)测定水产饲料中三聚氰胺含量的方法进行优化,得到最佳色谱条件和前处理方法,建立了水产饲料中三聚氰胺的检测方法。结果表明:以体积比8∶92的乙腈、庚烷磺酸钠离子对试剂为流动相,柱温40℃,流速1.2 mL/min,以波长236 nm检测;以提取液为18 mL50%(φ)乙腈水+1 mL乙酸锌+1 mL1mol/L盐酸,淋洗液3 mL水和3 mL甲醇,洗脱体积6 mL作样品前处理。三聚氰胺加标回收率为85%～100%,相对标准偏差(RSD)小于5%,线性范围为0.2 mg/L～5.0mg/L,相关系数0.999 8,方法检出限为0.2 mg/kg。     

Isolation and characterization of venom from nematocysts of jellyfish Rhopilema esculentum Kishinouye

The present work is the first report of the biochemical characterization of the venom from nematocysts of the jellyfish Rhopilema esculentum Kishinouye. The nematocysts were isolated by autolysis and centrifugation and separated by flow cytometry. Four types of nematocysts were identified: mastigophores, euryteles, and atrichous and holotrichous isorhiza. SDS-PAGE and amino acid analyses demonstrated that most of the proteins in the nematocyst extract were between 10 kDa and 40 kDa, and that glutamic acid was the main amino acid. A hemolytic activity assay showed that the activity of the nematocyst venom (RNV) was strongest in Tris-HCl buffer (50 mmol/L, pH 7.8, 5% glycerol, 0.5 mmol/L EDTA, 0.1 mol/L NaCl). The hemolytic activity was related to protein concentration and the HU50 against chicken erythrocytes was 0.91 μg/mL.     

Screening of extraction methods for glycoproteins from jellyfish (Rhopilema esculentum) oral-arms by high performance liquid chromatography

In order to select an optimum extraction method for the target glycoprotein (TGP) from jellyfish (Rhopilema esculentum) oral-arms, a high performance liquid chromatography (HPLC)-assay for the determination of the TGP was developed. Purified target glycoprotein was taken as a standard glycoprotein. The results showed that the calibration curves for peak area plotted against concentration for TGP were linear (r = 0.9984, y = 4.5895x+47.601) over concentrations ranging from 50 to 400 mgL-1. The mean extraction recovery was 97.84% (CV2.60%). The fractions containing TGP were isolated from jellyfish (R. esculentum) oral-arms by four extraction methods: 1) water extraction (WE), 2) phosphate buffer solution (PBS) extraction (PE), 3) ultrasound-assisted water extraction (UA-WE), 4) ultrasound-assisted PBS extraction (UA-PE). The lyophilized extract was dissolved in Milli-Q water and analyzed directly on a short TSK-GEL G4000PWXL (7.8 mm×300 mm) column. Our results indicated that the UA-PE method was the optimum extraction method selected by HPLC.     

SCREENING OF AGGLUTININS IN MARINE ALGAE FROM FUJIAN COAST OF CHINA

Thirty-three species of marine algae belonging to Rhodophyta, Phaeophyta and Chlorophyta from the Fujian coast were examined for agglutinins with different animal and human erythrocytes. Protein extracts from 26 species were active against at least one type of the erythrocytes tested. There were 3 species (Grateloupia imbricata, lshigefoliacea and Entermorpha prolifera) whose extracts could agglutinate all the erythrocytes used. The lowest protein concentration required to produce erythrocyte agglutination varied remarkably, from 3.1μg/ml to 500μg/ml . The strongest activity was found in the agglutina-tion of rabbit erythrocytes by Gloiopeltis furcata extract. Inhibition assays performed with nine mono- and bisaccharides indicated that agglutinations of rabbit erythrocytes by extracts of 7 species were inhibited by one or more types of the sugars assayed. The agglutinating activity shown by extracts of most species wasnot affected when the test solution was heated to 90℃, but was lost at 95℃ - 100℃. A few extracts losttheir activity at 60 RS, 65 RS and 75 RS, respectively.     

基于梯度对称性的无人机影像电力线自动提取方法

针对航空影像电力线提取过程中,背景线特征(电力杆塔、道路和植被等)干扰问题和电力线提取准确性问题,提出一种基于梯度对称性的电力线特征判别方法。采用Line Segment Detector(LSD)算法,对航空影像进行线特征提取;基于电力线附近梯度的对称性特征,实现从大量线段中准确筛选电力线段;采用缓冲区序列化线段连接算法,实现了电力线的自动连接。通过实验证明,提出的方法能够排除背景干扰并准确、完整地提取整条电力线。     

半叶马尾藻对N、P吸收速率的初步研究

在实验室条件下研究半叶马尾藻对不同浓度N、P以及不同化合态N的吸收速率。结果显示:不同浓度N、P下,半叶马尾藻对N、P的吸收速率差异有统计学意义(P0.05);半叶马尾藻对N、P的吸收速率随N、P浓度的升高而增加,最大吸收速率出现在N为200μmol/L、P为20μmol/L时,分别为2.397和0.229μmol·g-1·h-1;不同氮磷比条件下,半叶马尾藻对N、P的最大吸收速率出现在氮磷比为10时,在氮磷比为1~30范围内,氮磷比对半叶马尾藻吸收N、P速率的影响有统计学意义(P0.05);不同化合态N对半叶马尾藻吸收N、P速率的影响有统计学意义(P0.05),当NH4+-N和NO3--N共存时,半叶马尾藻对这2种化合态氮可同时吸收利用,各处理组间,对总无机N的吸收速率差异有统计学意义(P0.05),在c(NO3--N)/c(NH4+-N)为2∶1和1∶2处有两个峰值,分别为2.036、1.862μmol·g-1·h-1,对P的最大吸收速率均出现在c(NO3--N)/c(NH4+-N)值为2∶1时,为0.206μmol·g-1·h-1。因此,在粤西沿海半叶马尾藻在消除养殖水域营养盐、防治海水富营养化方面有很大优势。     

Growth inhibition to three red tide microalgae by extracts of Ulva pertusa

1 INTRODUCTION In recent decades, red tides that induced by in- creasing eutrophication occur frequently in coastal areas worldwide. This has caused serious problems in marine environment, aquaculture industries and human health (Qi et al., 1993; Horner e…     

Histological Characterization of Peppermint Shrimp(Lysmata wurdemanni) Androgenic Gland

The androgenic gland(AG) is an important endocrine gland for male reproductive function in crustaceans. In the present study, we investigated the histological characteristics of the androgenic gland of peppermint shrimp, Lysmata wurdemanni. The peppermint shrimp matures as male first, then some individuals may become euhermaphrodite after several moltings(transitional phase). Euhermaphrodite-phase shrimp acts as male at intermolts. However, it can be fertilized as a female immediately after molting. Considering the male reproductive function acts in its lifespan except for at larval stages, and female reproductive system starts to develop at transitional phase, we hypothesized that AG activity might be reduced to allow and promote vitellogenesis onset in early transitional phase and the following euhermaphrodite phase. So AG cell structure might be different in three phases in L. wurdemanni. The results showed that AG exists in the male in transitional and euhermaphrodite phases. The gland cell clusters surrounding the ejaculatory ducts locate at the roots of the fifth pereopods. The nucleus diameters are similar in the three phases while the nucleus-to-cell ratio is the lowest in euhermaphrodite phase. Our results indicated that for the individuals that will become euhermaphrodite, the cellular structure of AG changes since transitional phase. Male reproductive function which is still available in euhermaphrodite-phase shrimp should be due to the existence of the gland.