基于胃含物和稳定同位素研究海州湾长蛇鲻的摄食习性

长蛇鲻(Saurida elongate)为温水性近海底栖鱼类,是目前海州湾的优势鱼种之一,在海州湾食物网中占据重要地位。本研究基于海州湾底拖网调查采集的样品,结合胃含物分析和碳、氮稳定同位素技术,分析了长蛇鲻的摄食习性。结果表明: 海州湾长蛇鲻摄食的主要饵料生物为戴氏赤虾、枪乌贼、长丝虾虎鱼、鳀、六丝钝尾虾虎鱼、细条天竺鲷、尖海龙和绯鱼衔。其δ¹³C值范围在-19.39‰～-16.23‰之间,平均值为(-18.01±0.85)‰,与体长无显著相关性;δ¹⁵N值范围在9.56‰～13.36‰之间,平均值为(11.77±0.86)‰,与体长呈显著正相关。长蛇鲻不同体长组的饵料生物贡献度差异较大,随着体长增加,其捕食能力增强、摄食器官逐渐完善、摄食饵料生物的比例发生变化,可能是导致其摄食习性随体长变化的主要原因。本研究有助于深入了解海州湾长蛇鲻的摄食生态,为海州湾食物网营养动力学研究提供了基础资料。     

Root herbivore identity matters in plant-mediated interactions between root and shoot herbivores

Plants are simultaneously attacked by a multitude of herbivores that affect plant responses and plant-mediated interactions in a variety of ways. So far, studies on indirect interactions between below- and aboveground herbivores have almost exclusively focused on interactions between only one root and one shoot herbivore species at the same time. Since these studies show a variety of outcomes, we test the hypothesis that root herbivore identity matters in below-/aboveground interactions. We studied the combined effects root-feeding nematodes (Pratylenchus penetrans) and wireworms (Agriotes lineatus larvae) on Plantago lanceolata and on the performance of aboveground phloem-feeding aphids (Myzus persicae) and chewing caterpillars (Chrysodeixis chalcites larvae). Since root herbivores may also affect resource availability and the microbial community in the rhizosphere, we examined resource utilization by soil microorganisms using BIOLOG EcoPlates™.

Wireworms decreased root biomass by 13%, but led to compensatory shoot growth. Nematodes and the aboveground herbivores did not affect the biomass of Plantago lanceolata. Feeding by C. chalcites larvae enhanced the concentration of aucubin in leaves, which might explain the high mortality of the caterpillars. Aphids and the belowground herbivores did not change iridoid glycoside levels in the leaves. However, the number of aphid offspring was reduced by 44% when nematodes had been added to the soil, whereas wireworms had no effect. We observed higher utilization of BIOLOG carbon sources by the soil microorganisms only in the presence of Pratylenchus penetrans. Our results suggest that the outcome of below–aboveground interactions highly depends on herbivore identity.     

Validation of internal controls for gene expression analysis in the intestine of rats infected with Hymenolepis diminuta

The non-invasive parasitic cestode Hymenolepis diminuta induces hypertrophy, hyperplasia and other changes in cell activity in the intestine of rats which are indicated in the expression of mRNA. We have investigated various house-keeping genes (GAPDH, β-actin, 18S and HPRT) and other internal controls (total RNA/unit biomass, total RNA/unit length of intestine) to validate gene expression in the rat intestine after cestode infection and drug-induced neuromodulation. Variation in GAPDH, β-actin, 18S and HPRT expression was observed in rat jejunal tissue according to treatment. Total RNA/unit length of intestine was found to be the most suitable internal control for normalizing target gene mRNA expression in both infected and/or drug-induced rat intestine. This normalization method may be applied to studies of gene expression levels in intestinal tissue where hypertrophy, hyperplasia, rapid growth and cell differentiation generally occur.     

Structures of the galactomannans from seeds of Annona muricata, Arenga saccharifera, Cocos nucifera, Convolvulus tricolor, and Sophora japonica

Galactomannans were isolated from ripe seeds of Annona muricata (Annonaceae), Convolvulus tricolor (Convolvulaceae), Sophora japonica (Leguminosae), and from immature seeds of Arenga saccharifera and Cocos nucifera (both Palmae). Their sugar compositions were determined and their structures studied by the methylation and periodate-oxidation techniques. All the galactomannans studied are of the leguminous type, the molecules having main chains consisting of (1→4)-linked β- -mannose residues, with differing proportions of side chains consisting of single - -galactose residues linked to the main chains by (1→6)-bonds. The molecular weights were found to vary from 6,000 (Sophora) to 17,000 (Arenga). The isolation of the galactomannan of Annona is the first recorded occurrence of this type of polysaccharide in the family Annonaceae, whereas there has been a previous report of the occurrence of a galactomannan in the Convolvulaceae; the study of the structure of the Sophora galactomannan is the first one in the tribe Sophoreae of the Leguminosae.     

Coproantigen detection in dogs experimentally and naturally infected with Echinococcus granulosus by a monoclonal antibody-based enzyme-linked immunosorbent assay

A sandwich ELISA for the detection of Echinococcus granulosus coproantigen in formalin and heat-treated faecal supernatants of dogs was developed. The assay used affinity-purified polyclonal antibodies obtained from rabbits hyperimmunised with E. granulosus excretory/secretory antigens and biotiaylated monoclonal antibody EmA9 produced against adult E. multilocularis somatic extract. The test was sensitive to 7 ng and 2.3 ng of E. granulosus protein and carbohydrate/ml of faecal supernatant, respectively. Thirteen helminth-free dogs were infected with different amounts of E. granulosus protoscoleces and the presence of coproantigen was monitored during the prepatent period until day 35 post-infection, when they were necropsied. Faecal antigen levels started to rise above the normal range between days 10 and 20 post-infection, and typically peaked at the end of the experiment. All the dogs, bearing from 3 to 67 700 worms, showed positive values in the ELISA during the prepatent period. One dog experimentally infected with Taenia hydatigena metacestode and harbouring three worms, tested positive only after the prepatent period at day 52. The test was applied to 98 stray dogs. The ELISA detected all of four dogs naturally infected with E. granulosus, two dogs with patent infections of T. hydatigena and two dogs with no cestode infections, sbowing a sensitivity of 100% and a specificity of 96%.     

Anthelmintic efficacy of Flemingia vestita (Fabaceae): alterations in glucose metabolism of the cestode, Raillietina echinobothrida

The root-tuber peel of Flemingia vestita and its active component, genistein, were tested in respect of glucose metabolism in the cestode, Raillietina echinobothrida. Live R. echinobothrida, collected from the intestine of freshly slaughtered domestic fowl, were incubated at 39±1 °C in defined concentrations of the root-peel crude extract (5 mg/ml), genistein (0.2 mg/ml) and praziquantel (1 μg/ml) in phosphate buffered saline with 1% of dimethyl sulphoxide with simultaneous maintenance of controls. In the treated worms, there was a significant decrease in the glycogen concentration accompanied with the decrease of glucose by 14–32%, whereas the malate concentration increased by 49–134% as compared to controls. Both in controls and treated parasites, however, the pyruvate content was not measurable. While alanine and lactate contents showed a decline by 7–25% in the parasites exposed to all test materials, the lactate efflux into the incubation medium showed 37–71% increase in treatments indicating an overall increase of lactate production in comparison to controls. The results showing a decline in the glycogen and glucose contents and a significant rise in the malate content and lactate efflux under treatment conditions suggest that the energy demand in the parasites possibly got enhanced under stress, though it did not influence a switch over towards aerobic degradation of glucose in the parasites.     

紫陀螺菌化学成分及其抑制肿瘤细胞增殖活性的研究

以紫陀螺菌为对象,研究其子实体的化学成分及其抑制肿瘤细胞增殖活性。采用溶剂提取、柱层析和高效液相色谱等方法分离纯化化学成分,通过核磁共振和质谱技术鉴定单体化合物结构,运用结晶紫法评价单体化合物抑制肿瘤细胞增殖活性。从乙酸乙酯提取物中共分离鉴定6个单体化合物,分别为(22E,24R)-麦角甾-5,7,22-三烯-3β-醇(1)、3β,5α-二羟基-(22E,24R)-麦角甾-7,22-二烯-6-酮(2)、(22E,24R)-麦角甾-7,22-二烯-3β,5α,6β-三醇(3)、吲哚-3-甲酸甲酯(4)、4,4-二甲基-1,7-庚二酸(5)和(8E,10E)-12羰基十八碳-8,10-二烯酸(6),其中化合物1为主要成分,相对含量为23.8%。活性测试结果表明3对人乳腺癌细胞株MCF-7 细胞、人胰腺癌细胞株PANC-1细胞和人乳腺癌细胞株MDA-MB-231细胞具有微弱的细胞增殖抑制活性。本研究首次报道了紫陀螺菌化学成分,对深入挖掘其在健康领域中的开发价值具有重要意义。     

海洋真菌Penicillium sp. WP-13次级代谢产物研究

本研究分析了海洋真菌Penicillium sp. WP-13的活性次级代谢产物。采用多种柱色谱技术对其发酵液的乙酸乙酯提取物进行分离纯化;根据波谱数据和理化常数分析,并结合文献比对鉴定单体化合物的结构;采用MTT法测定化合物对肿瘤细胞的细胞毒活性。从Penicillium sp. WP-13发酵液的乙酸乙酯提取物中分离鉴定了5个单体化合物,其中2个内酯类化合物为1-hydroxy-3,10-dimethoxy-8-methyl-6,11-dioxo-6,11-dihydrodibenzo[b,e]oxepine-9-carboxylicacid(1)和1,8-dihydroxy-10-methoxy-3-methyl-dibenzo[b,e]oxepine-6,11-dione(2); 3个蒽醌类化合物为endocrocinmethylester(3)、2-chloro-1,3,8-trihydroxy-6-methyl-9,10-anthracenedione (4)和emodin (5)。活性测试结果显示,化合物3和5均对人慢性髓原白血病细胞株K562、人肝癌细胞株BEL-740...     

Efficiency of procedures for induction and cultivation of Pseudomonas syringae pv. pisi L-form

The L-form of Pseudomonas syringae pv. phaseolicola has been proved to induce resistance to bean halo blight.Various procedures were tested to induce the L-form of Pseudomonas syringae pv. pisi for its potential use as biocontrol agent of pea bacterial blight. Cell-wall deficient cells were induced in a liquid medium with penicillin following a protocol described for P. s. pv. phaseolicola. Cell growth on solid induction medium developed as typical granular and vacuolated structures, and characteristic colonies were observed in the first transfer. However, there was poor growth in subsequent transfers and some reversion to the parental type. To improve the induction procedure, the following new procedures were applied: (1) viability of cells was monitored during induction. The optimum induction time in liquid medium with penicillin was lower for pv. pisi than for pv. phaseolicola. Viability of L-forms in solid induction medium with penicillin was low and decreased in time. (2) the inducer ticarcillin was combined with clavulanic acid, which prevented the reversion to the parental type and (3) a range of concentrations of penicillin and ticarcillin/clavulanic acid was applied by the spiral gradient endpoint method for calculation of minimum inhibitory concentrations (MIC). Based on the results from these tests an induction method for P. s. pv. pisi L-form is proposed and the relevance of L-form is discussed for practice.     

Ergosterol peroxides as phospholipase A2 inhibitors from the fungus Lactarius hatsudake

Four ergosterol derivatives (1–4) have been isolated for the first time from the fruiting bodies of a basidiomycete fungus, Lactarius hatsudake, through activity-guided fractionation. Their structures were determined, using spectroscopic analysis, as: (22E,24R)-ergosta-5,7,22-dien-3β-ol (ergosterol, 1); 5,8-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol (ergosterol peroxide, 2); 5,8-epidioxy-(24S)-ergosta-6-en-3β-ol (3); and (22E,24R)-ergosta-7,22-dien-3β,5,6β-triol (cerevisterol, 4). Compounds 2 and 3 showed selective inhibitory activity against Crotalus adamenteus venom phospholipase A₂ (PLA₂) enzyme, but not against Apis mellifcra bee venom PLA₂. The antiphospholipase A₂ activity of compounds 2 and 3 are reported here for the first time.     

山东对囊蕨属(蹄盖蕨科)植物孢粉学研究及其在分类上的意义

采用扫描电镜对山东分布的8种对囊蕨属(Deparia)植物进行孢粉学观察。研究结果表明:在中国对对囊蕨属(Deparia Hook.& Grev.)蹄盖蕨科(Athyriaceae)新分类系统首次进行孢子形态亚显微结构研究;该属孢子形态圆肾形,左右对称,具周壁,其周壁纹饰在种内稳定,种间区别显著;为建立假蹄盖蕨亚属(Subgen.1.Athyriopsis)和蛾眉蕨亚属(Subgen.2.Lunathyrium )提供孢粉学依据;依据山东蛾眉蕨和东北蛾眉蕨孢壁纹饰的显著差异,山东蛾眉蕨应为一个独立的物种,不宜并入东北蛾眉蕨,建议恢复山东蛾眉蕨在植物分类学上的种级地位,依照对囊蕨属新分类系统,山东蛾眉蕨(Lunathyrium shandongense)新组合为中华山东对囊蕨(D.sinoshandongensis)。该研究不仅为对囊蕨属孢粉学积累了新的资料,也为近缘种的分类鉴定提供了孢粉学依据。因此对囊蕨属孢子形态研究,在该属孢粉学、分类学及系统演化上都具有重要意义。     

First identification of eggs of the Asian fish tapeworm Bothriocephalus acheilognathi (Cestoda: Bothriocephalidea) in human stool

We report the first case of egg isolation of the Asian fish tapeworm Bothriocephalus acheilognathi (Bothriocephalidea) from human stool. A male patient from Saint Laurent du Maroni (French Guiana) presenting abdominal pain was examined in France for the diagnosis of intestinal parasites. Diphyllobothrium-like eggs were observed in his stool. However, molecular phylogenetic analyses based on sequences of rDNA and COI genes showed that the eggs observed belong to a bothriocephalidean cestode B. acheilognathi. The adult life stages of B. acheilognathi cestodes are known as invasive parasites of a wide spectrum of fish; however, they have not been described to parasitize any mammals. This human infection seems to be accidental and represents a parasite passage through human intestine after the consumption of an infected fish host.     

六妹羊肚菌的化学成分

以六妹羊肚菌Morchella sextelata为研究对象,对其子实体的化学成分进行研究。采用气相色谱-质谱联用技术（GC-MS）进行子实体芳香物和亲脂性提取物的化学成分分析,同时对其亲脂性提取物的抗氧化和抗菌活性进行了初步评价;采用正反相硅胶柱色谱、葡聚糖凝胶柱色谱等多种色谱分离方法进行化学成分的分离纯化,并通过核磁共振（NMR）、质谱（MS）等技术鉴定化合物结构。从六妹羊肚菌子实体的芳香物中共鉴定出26个化合物,辛-1-烯-3-醇（32.53%）、(E)-辛-2-烯醛（25.15%）和苯乙醛（12.31%）为主要成分;从六妹羊肚菌子实体的亲脂性提取物中共鉴定出14个化合物,亚油酸（77.80%）为主要成分;六妹羊肚菌亲脂性提取物仅显示出中等强度的抗氧化活性。从六妹羊肚菌子实体中共分离鉴定出14个化合物,包括7个甾体类化合物,其中化合物1、2、4、7、11、13和14为首次从羊肚菌属中分离得到。本研究首次对六妹羊肚菌的小分子化学成分进行了分析,对羊肚菌活性物质的阐明及进一步开发利用具有重要意义。     

舌瓣鼠尾草退化杠杆雄蕊的相关花部特征及传粉机制

杠杆状雄蕊是鼠尾草属(Salvia)物种形成的关键性状, 背部杠杆传粉模式作为该属植物与传粉者精确互作的经典案例被广泛深入研究, 但是在该属物种中还存在许多非典型的杠杆结构和传粉模式。雄蕊结构及其与传粉者互作的多样性, 使得鼠尾草属成为研究植物传粉模式转变的模式材料, 舌瓣鼠尾草(S. liguliloba)即是一种具非典型的退化杠杆状雄蕊结构和传粉特征的代表性物种。该文着重对舌瓣鼠尾草的花器官结构和传粉特征进行研究, 并与具有短药隔杠杆的毛地黄鼠尾草 (S. digitaloides)做比较分析, 以期揭示退化杠杆可能的进化选择压力及其生态学意义。结果表明, 舌瓣鼠尾草具有较短的花冠、更窄的冠筒和较短的雄、雌蕊(p < 0.05)。退化萎缩的雄蕊下臂, 冠筒内的狭小空间限制了唯一的有效传粉昆虫——三条熊蜂(Bombus trifasciatus)推动雄蕊做杠杆状运动, 而是靠近花药直接利用头部完成授粉。相比经典的杠杆状雄蕊结构及其传粉过程, 小型花冠和退化杠杆雄蕊是对专一性和活跃度较高传粉昆虫的适应, 可能具有完全不同的进化途径和繁殖策略。     

Echinococcal cyst of the subcutaneous tissue: a rare case report

Hydatid disease is an endemic zoonosis caused by the cestode Echinococcus granulosus. The most common affected organs are the liver and the lungs. Echinococcal cyst located in the subcutaneous tissue constitutes an extremely rare manifestation of the hydatid disease. Here we present a case report with a cystic mass in the subcutaneous tissue of the left gluteal area, diagnosed as being a hydatid cyst.     

Insecticidal lead identification by screening benzopyrano[4,3-c] pyrazol-3(2H)-ones library constructed from multiple-parallel synthesis under microwave irradiation

A rapid library-generation via liquid-phase multiple-parallel synthesis of 2-(substituted)benzyl-1-benzopyrano[4,3-c]pyrazol-3(2H)-ones, bearing two points of diversity, under microwave irradiation was successfully performed using chromenone-3-carboxylic acids as starting materials. Compared to an identical library generated by conventional parallel synthesis, microwave-assisted parallel synthesis dramatically decreased reaction times from an average of 16 h to 13 min, and the yields of products and intermediates were improved in most cases. A bioassay indicated that the compounds of the library exhibited excellent insecticidal activity against T. cinnabarinus at the dosage of 250 mg L⁻¹, and some compounds still exhibited insecticidal activity when the dosage was reduced to 50 mg L⁻¹. This shows that 2-(substituted) benzyl-1-benzopyrano[4,3-c]pyrazol-3(2H)-ones might be used as lead structures for further optimization. To our knowledge, this is the first report about the application of solution-phase multiple-parallel synthesis under microwave irradiation to construct libraries of benzopyrano-[4,3-c]pyrazol-3(2H)-ones with insecticidal activity. The coupling of microwave technology with liquid-phase parallel synthesis constitutes a novel and particularly attractive avenue for the rapid generation of structurally diverse libraries for lead discovery.     

Characterisation of sorbed water molecules on neutral and ionic polysaccharides

Ionic and neutral polysaccharides with well-defined structures were chosen to investigate the mechanism of water sorption at different relative humidities. From an experimental point of view, the freezing water was determined by DSC when the total sorbed water was obtained from thermogravimetry. The isotherms of sorption and enthalpies of interaction were determined using the combination of a microbalance and a microcalorimeter. It is shown that freezing water appears for P/P₀ > 0.85 especially with the neutral polymers. The differential molar enthalpy of interaction is higher for P/P₀ < 0.85 corresponding to the fixation of two water molecules forming double H-bonds; this result is confirmed by molecular modelling; saturation is obtained experimentally for 4 water molecules interacting per glucose unit. On ionic polymers, the water retention increases especially over P/P₀ 0.8 and the enthalpy of interaction is higher for the first water molecules sorbed. For P/P₀ 0.8, the numbers of bound water molecules found are 2 per glucopyranosyl unit for neutral polysaccharides, 5 for glucuronan and 9–10 for carboxymethylcellulose (CMC) of D = 2 and hyaluronan (HA)     

Echinococcus multilocularis in a European beaver from Switzerland

Infection with the larval stage of the cestode parasite Echinococcus multilocularis was diagnosed in a European beaver (Castor fiber) in central Switzerland. The animal was hit, run over by a car, and died of trauma. It was in normal body condition and no signs of disease were seen. At necropsy, multiple cystic structures up to 1 cm in diameter were found in the liver adjacent to the hilus. Within the parasite vesicles, multiple protoscolices were visible. The species was determined to be Echinococcus multilocularis by upon polymerase chain reaction and direct immunofluorescence with MAbG11-FITC. This is the first report of Echinococcus multilocularis in European beaver.     

Mutation of fungal endoglucanases into glycosynthases and characterization of their acceptor substrate specificity

Humicola insolens mutant Cel7B E197A is a powerful endo-glycosynthase displaying an acceptor substrate specificity restricted to β-d-glucosyl, β-d-xylosyl, β-d-mannosyl and β-d-glucosaminyl in +1 subsite. Our aim was to extend this substrate specificity to β-d-N-acetylglucosaminyl, in order to get access to a wider array of oligosaccharidic structures obtained through glycosynthase assisted synthesis. In a first approach a trisaccharide bearing a β-d-N-acetylglucosaminyl residue was docked at the +1 subsite of H. insolens Cel7B, indicating that the mutation of only one residue, His209, could lead to the expected wider acceptor specificity. Three H. insolens Cel7B glycosynthase mutants (H209A, H209G and H209A/A211T) were produced and expressed in Aspergillus oryzae. In parallel, sequence alignment investigations showed that several cellulases from family GH7 display an alanine residue instead of histidine at position 209. Amongst them, Trichoderma reesei Cel7B, an endoglucanase sharing the highest degree of sequence identity with Humicola Cel7B, was found to naturally accept a β-d-N-acetylglucosaminyl residue at +1 subsite. The T. reesei Cel7B mutant nucleophile E196A was produced and expressed in Saccharomyces cerevisiae, and its activity as glycosynthase, together with the H. insolens glycosynthase mutants, was evaluated toward various glycosidic acceptors.