产黄青霉Penicillium chrysogenum S-3-25人工海水培养基发酵次级代谢产物研究

目的 阐明南极深海来源真菌产黄青霉Penicillium chrysogenum S-3-25人工海水培养基的发酵次级代谢产物及其活性。方法 利用各种色谱技术分离纯化次级代谢产物,根据理化和波谱数据（核磁共振、质谱技术和Marfey分析）鉴定化合物结构,采用3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide（MTT）法评价细胞毒活性。结果 从真菌S-3-25人工海水培养基发酵产物中分离鉴定了8个单体化合物：2-[[(2S)-2-amino-1-oxopropyl] amino] benzoic acid (1),methyl 2-[[(2S)-2-amino-1-oxopropyl] amino] benzoate (2),2-[[(2S)-2-hydroxy-1-oxopropyl] amino] benzamide (3),3-(p-hydroxyphenyl)-N-methylpropionamide (4),4-hydroxycinnamamide (5),cyclo-(L-Pro-L-Tyr) (6),脑苷脂A (7) 及脑苷脂B (8)。细胞毒活性测试结果表明化合物1~8在50 μM作用浓度下对三种受试细胞（人乳腺癌MCF-7,人肺癌A549以及小鼠小胶质BV2细胞）的抑制率均低于50%。结论 从极地深海来源真菌产黄青霉S-3-25人工海水培养基发酵产物中分离得到8个单体化合物,其中化合物1和2为新天然产物,未见有文献数据的报道。化合物1~8均未呈现较强的细胞毒活性。     

深海真菌Phomopsis tersa的次级代谢产物及其细胞毒活性研究

目的 研究深海真菌Phomopsis tersa FS441的次级代谢产物及其细胞毒活性。方法 采用正、反相硅胶柱、凝胶柱和高效液相等多种色谱技术对菌株FS441的固体发酵产物进行分离纯化,并通过NMR、MS等波谱方法鉴定化合物结构,采用SRB法对得到的单体化合物进行细胞毒活性测试。结果 从发酵物中分离纯化得到11个化合物,分别鉴定为：5α, 8α-表二氧麦角甾-6,22-二烯-3β-醇（1）、麦角甾醇（2）、12-hydroxydehydrobotrydienol（3）、5,6-二羟基-2,3,6-三甲基环己-2-烯酮（4）、1,2-二羟基-3,4-脱氢蒽酮（5）、（1S,2S,3S,4R） -3-氯-4-（2-羟基丙吡啶-2-基）-1-甲基环己烷-1,2-二醇（6）、（-）-5-methylmellein（7）、酪醇（8）、胸腺嘧啶核苷（9）、（R）-（+）-甘油1-9 （Z）,12 （Z）-十八碳二烯酸酯（10）、亚油酸（11）。结论 化合物3～6为首次从Phomopsis属分离得到,化合物1和2对SF-268、MCF-7、HepG-2和A549 4种肿瘤细胞株具有弱的细胞毒活性。     

北极真菌Nectria sp.B-13中酚醛倍半萜烯类化合物的研究

摘要：目的 对北极真菌 Nectria sp. B-13 中的酚醛倍半萜烯类化合物进行结构鉴定及生物学活性研究。方法 采用硅胶柱层析、Sephadex LH-20凝胶柱层析、高效液相色谱等分离手段对极地真菌Nectria sp. B-13发酵液的乙酸乙酯萃取部位进行了系统的分离,利用1H-和13C-NMR、LC-MS等现代波谱解析手段,结合文献对照,鉴定化合物结构;采用MTT法和微量稀释法测试了化合物的细胞毒活性和抗稻瘟霉作用。结果 从提取物中分离得到6个化合物,其中4个为酚醛倍半萜烯类化合物,分别为dechlorodeacetylchloronectrin（1）,LL-Z-1272 ε acid（2）,8''-hydroxyascochlorin（3）和LL-Z 1272 β（4）,以及邻苯二甲酸丁酯异丁酯（5）和邻苯二甲酸二丁酯（6）。活性测试的结果表明,化合物2对5种肿瘤细胞株均有一定程度的生长抑制作用,其IC50值的范围在48.67~92.18 μM之间;同时,化合物2还可抑制稻瘟霉的生长,MIC值为18.2 μg/mL。结论 化合物1和化合物2为首次从微生物次级代谢产物中获得的化合物,为新天然产物。化合物2有一定的细胞毒活性和抗稻瘟霉作用。     

1株老鼠簕内生真菌Trichoderma sp. RSF-1的次级代谢产物分离及生物学活性研究

目的 对漳江口红树林植物老鼠簕（Acanthus ilicifolius）内生真菌Trichoderma sp. RSF-1的次级代谢产物和生物活性进行研究。方法 综合利用硅胶柱色谱、凝胶柱层析及高效液相色谱（HPLC）等分离手段对菌株发酵液粗提物进行次级代谢产物的分离纯化;利用现代波谱学方法（核磁共振、质谱及旋光等）对单体化合物进行结构鉴定;对所得化合物进行细胞毒活性评价和抗菌活性筛选。结果 从Trichoderma sp. RSF-1大米发酵产物中分离鉴定了13个单体化合物：包括1个丁内酯化合物1,5个聚酮类化合物2~6,1个生物碱类化合物7,2个二倍半萜类化合物8和9,3个甾体类化合物10~12及1个脂肪酸类化合物13：piterrione Ⅰ（1）、nafuredin（2）、penicilloxalone A（3）、5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone（4）、5-hydroxy-7-methoxy-2,3-dimethyl-4H-chromen-4-one（5）、fusarimone A（6）、(3R,6R)-4-methyl-6-(1-methylethyl)-3-phenylmethylperhydro-1,4-oxazine-2,5-dione（7）、fusaproliferin（8）、terpestacin（9）、麦角甾醇（10）、β-谷甾醇（11）、啤酒甾醇（12）及亚油酸（13）。结论 从漳江口红树林植物老鼠簕内生真菌Trichoderma sp. RSF-1中分离获得了化合物1~13,其中化合物1为新化合物,7、8和9首次从该种属分离得到,化合物7对多种癌细胞系均有显著的细胞毒活性,化合物8对部分癌细胞具有中等的细胞毒活性。化合物7和13对多种致病菌包括枯草芽孢杆菌、杀鲑气单胞菌和鳗弧菌具有较强抑制作用,其最小抑菌浓度（MIC）值为6.25~50.00 μg/mL。     

2株深海来源真菌次级代谢产物的研究D

目的 对2株深海来源真菌Graphostroma sp. MCCC 3A00421和Aspergillus sp. MCCC 3A400414进行化学成分和生物活性研究。方法 采用溶剂萃取、柱色谱层析及半制备HPLC等方法对2株深海真菌的发酵产物进行化学分离,通过理化性质及波谱学方法并参阅文献进行化合物结构鉴定,并初步评价其细胞毒活性。结果 从菌株3A00421的发酵产物中分离得到了4个化合物,经鉴定其结构分别为3,5-dimethyl-8-methoxy-3,4-dihydroisocoumarin（1）、6-O-methylreticulol（2）、nectriapyrone（3）、demethyl nectriapyrone A（4）;从菌株3A00414中分离得到了5个化合物,经鉴定其结构分别为(22E,24R) -麦角甾-5,7,22-三烯-3β-醇（5）、WIN 64821（6）、diorcinol（7）、9-十八碳烯酸（8）、9,12-十八碳二烯酸（9）。其中化合物6具有中等强度的细胞毒活性,其IC50为7.14～15.78 μmol/L。结论 深海真菌是重要的活性物质来源和生物资源,本研究首次从深海来源Graphostroma属真菌的代谢产物中发现了2个异香豆素和2个苯并吡喃酮类化合物。     

角果木内生真菌Coriolopsis sp. J5代谢产物及活性研究

摘 要：目的 研究红树植物角果木内生真菌Coriolopsis sp. J5的活性次生代谢产物。方法 采用多种柱色谱技术对次生代谢产物进行分离纯化,根据理化常数和波谱数据分析鉴定化合物的结构;分别采用MTT法和滤纸片琼脂扩散法测定化合物的体外细胞毒活性及抗菌活性。结果 从角果木内生真菌Coriolopsis sp. J5的次生代谢产物中分离鉴定了7个化合物,分别为14-acetoxy-15-hydroxyirpexan（1）、15α-hydroxytrametenolic acid（2）、3β-hydroxylanosta-8,24- dien-21-oic acid（3）、latifolicinin C（4）、对羟基苯乙酸甲酯（5）、methyl 5-（2-methoxycarbonylethy）furan-2-carboxylate（6）和5-(2-carboxy-ethyl)-furan-2-carboxylic acid（7）。化合物1对人慢性髓原白血病细胞(K-562)和人肝癌细胞（BEL-7420）生长具有抑制作用,化合物5具有抗白色念珠菌（Candida albicans）、棉花黄萎病菌（Verticillium dahliae Kleb）和棉花枯萎镰刀菌（Fusarium oxysporum f.sp. vasinfectum）活性,化合物6具有抗地毯草黄单胞菌（Xanthomonas axonopodis）活性。结论 化合物1~5为首次从角果木内生真菌的次级代谢产物中分离得到,化合物6和7为新天然产物,并首次报道了化合物7的核磁数据;菌株Coriolopsis sp. J5能代谢产生具有细胞毒活性和抗菌活性的化合物。     

红树林植物内生真菌Stachybotrys chartarum HDN16-358次级代谢产物研究

目的 对红树林植物内生真菌Stachybotrys chartarum（HDN16-358）进行次级代谢产物的研究。方法 利用薄层色谱层析（TLC）,反相ODS,Sephadex LH-20、半制备型HPLC等色谱学方法对菌株发酵粗提物进行分离纯化;利用NMR、MS、UV等波谱学技术手段,解析确定化合物的结构,并对新天然产物1进行了细胞毒活性筛选。结果 分离鉴定了5个单体化合物：(2S)-5-Hydroxy-2,7-dimethyl-2-(4,8-dimethyl-3E,7-nonadienyl)-2H-chromene-6-carbaldehyde (1),BR-011 (2),LL-Z 1272β (3) and stachybotrylactone (4),5-Methylbenzene-1,3-diol (5)。结论 红树林来源真菌是重要活性物质来源和生物资源。从红树林植物内生真菌Stachybotrys chartarum中分离得化合物1-5,化合物1首次从自然界中分离得到,且具有中等细胞毒活性。     

深海来源的微生物抗肿瘤活性筛选及次级代谢产物的初步研究

目的 对来自深海的海水、海泥样品进行了微生物分离并通过抗肿瘤活性筛选获得活性菌株,并研究活性菌株c2b的次级代谢产物.方法 从样品中选择性分离得到真菌,并采用海虾生物致死法和人体慢性艇性白血病细胞(K562)为筛选模型对分离得到真菌的发酵产物进行抗肿瘤活性筛选;采用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段对c2b菌株发酵产物的活性部位进行了活性追踪分离,通过理化性质及渡谱学手段进行化学结构鉴定,以SRB法评价了化合物的抗肿瘤活性.结果与结论 从深海来源的样品中共分离获得29株真菌,其中7株具有细胞毒活性;从c2b活性菌株的发酵产物中分离得到6个单体化合物(1～6),其化学结构分别鉴定为N-乙酰色氨(1),chrysogine(2),过氧化麦角甾醇(3),5,8-epidioxy-24-methylcholesta-6,22-dien-3β-ol(4),cerevisterol(5)和(4E,8E)-N-[(2'R,3'E)-2'-hydroxy-3'-hexadecenoyl]-1-O-β-D-glycopyranosyl-9-methyl-4,8-sphingadiene(6),其中化合物3,4对小鼠乳腺癌细胞(tsFT210)具有中等强度的细胞毒活性.     

毛蚶共附生Aspergillus clavatonanicus JL001含氮类次级代谢产物的研究

目的 研究毛蚶共附生Aspergillus clavatonanicus JL001的含氮类次级代谢产物,并探索其细胞毒活性。方法 静置发酵后,萃取得到粗提物。采用正反相硅胶色谱法、半制备高效液相、制备薄层色谱法（TLC ）分离纯化真菌的次级代谢产物,根据化合物NMR和HRMS数据结合文献报道确定其结构。利用CCK8法测定次级代谢产物1～5对人肿瘤细胞HepG2的细胞毒活性。结果 从毛蚶共附生Aspergillus clavatonanicus JL001中分离得到了8个含氮类化合物,分别为tryptoquivaline L（1）、quinadoline A（2）、scequinadoline I（3）、prelapatin B（4）、clavatustide B（5）、环（(苯丙-丙)二肽（6）、环（(酪-亮)）二肽（7）和环（(酪-异亮)）二肽（8）。其中化合物3和4对HepG2细胞具有一定的增殖抑制作用。结论 研究结果为从海洋动物共附生真菌中寻找新型抗生素提供参考和依据。     

南极真菌Penicillium sp. S-2-10次级代谢产物的研究

目的 对来源于南极海域海底沉积物（47.09° W,62.05° S,水深1393 m）真菌Penicillium sp. S-2-10的次级代谢产物进行研究。方法 对菌株进行规模发酵,并对所得发酵液的乙酸乙酯萃取浸膏进行系统分离纯化,据化学指征和生物学活性两方面特点,采用Sephadex LH 20凝胶色谱、正相硅胶柱色谱、反相ODS柱色谱、高效液相柱层析等色谱分析方法,通过核磁（1H-NMR和13C-NMR等）及质谱（MS）等波谱分析方法,并与相关文献比较,鉴定化合物结构。 结果 分离得到7个单体化合物,经鉴定化合物1~7分别为Benzoic acid, 2-(2,6-dihydroxybenzoyl)-3-hydroxy-5-(hydroxymethyl)-methyl, methyl ester (1), chrysophanol (2), Volemolide (3), Ergosta-6,8(14),22-trien-3-ol (4), 2-aminophenoxazin-3-one (5), 2-benzothiazolol (6), benzoic acid (7)。结论 化合物3,4为首次从Penicillium属真菌中获得。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.