海洋放线菌Streptomyces sp.（No.30701）次生代谢产物研究

目的 研究一株海洋放线菌Streptomyces sp.（No.30701）的化学成分。方法 采用硅胶开放柱色谱、Sephadex LH-20凝胶柱色谱以及高效液相色谱等分离手段进行化合物的分离、纯化;利用理化性质和波谱学分析对单体化合物进行结构鉴定。结果 从海洋放线菌Streptomyces sp.（No.30701）发酵物中分离得到9个环二肽类化合物,分别鉴定为环(L-脯-L-缬)二肽（1）、环(D-脯-L-缬)二肽（2）、环(L-脯-L-亮)二肽（3）、环(4-羟基-脯-亮)二肽（4）、环(L-脯-L-异亮)二肽（5）、环(L-脯-L-酪)二肽（6）、环(L-亮-L-缬)二肽（7）、环(D-苯丙-甘)二肽（8）、环(L-苯丙-L-缬)二肽（9）。结论 以上化合物均为海洋放线菌常见的次生代谢产物,但化合物2、8含有天然界中不多见的D型氨基酸,并且化合物2是从链霉菌属放线菌中首次分离得到。     

海洋来源放线菌Demequina litorisediminis SCSIO 53428的化学成分研究

目的对南海深海沉积物来源的放线菌Demequina litorisediminis SCSIO 53428的发酵物化学成分进行研究。方法利用各种色谱技术,包括硅胶柱色谱、ODS反相柱色谱、Sephadex LH-20凝胶色谱、高效液相色谱等分离手段对放线菌Demequina litorisediminis发酵物的乙酸乙酯提取物进行分离纯化,通过波谱学数据分析并结合文献比较鉴定了化合物的结构。结果分离得到13个化合物,其结构分别为:环(L-酪-L-脯)二肽(1)、环(L-苯丙-L-脯)二肽(2)、环(L-缬-L-脯)二肽(3)、环(L-亮-L-脯)二肽(4)、环(反式-4-羟基-L-脯-L-亮)二肽(5)、环(甘-L-脯)二肽(6)、环(L-丙-L-脯)二肽(7)、环(缬-酪)二肽(8)、环(异亮-脯)二肽(9)、对羟基苯甲醛(10)、苯甲酸(11)、N-(2-羟基苯基)-乙酰胺(12)、2-乙酰氨基苯甲酸(13)。结论对海洋来源放线菌Demequina litorisediminis SCSIO53428的化学成分做了初步研究,化合物1～13均为首次从该属放线菌中分离得到。     

红榄李内生真菌Aspergillus terreus HT-1二酮哌嗪类次级代谢产物研究

目的 研究红榄李Lumnitzera littorea内生真菌Aspergillus terreus HT-1的次级代谢产物。方法 对从红榄李中分离得到的内生真菌A. terreus HT-1进行大规模发酵,并采用多种色谱技术对其发酵产物进行分离纯化。通过多种波谱方法鉴定单体化合物结构。结果 从A. terreus HT-1发酵产物的乙酸乙酯萃取物中分离得到了15个二酮哌嗪类化合物,分别鉴定为bis(dethio)bis(methylsulfanyl) gliotoxin（1）,环(D-4-羟基-脯氨酸-L-苯丙氨酸)二肽（2）,环(L-4-羟基-脯氨酸-L-苯丙氨酸)二肽（3）,环(L-4-羟基-脯氨酸-L-酪氨酸)二肽（4）,环(L-4-羟基-脯氨酸-L-亮氨酸)二肽（5）,环(L-丙氨酸-L-4-羟基-脯氨酸)二肽（6）,环(D-4-羟基-脯氨酸-D-异亮氨酸)二肽（7）,环(D-脯氨酸-L-酪氨酸)二肽（8）,环(L-亮氨酸-L-脯氨酸)二肽（9）,环(L-脯氨酸-L-苏氨酸)二肽（10）,环(L-脯氨酸-L-丙氨酸)二肽（11）,环(L-脯氨酸-甘氨酸)二肽（12）,环(L-苯丙氨酸-甘氨酸)二肽（13）,环(谷氨酸-L-酪氨酸)二肽（14）,terezine D（15）,并采用MTT法检测全部化合物的神经保护活性。结论 在200 μmol/L浓度下,化合物1和5显示出较好的神经保护活性,使H2O2诱导氧化损伤的神经细胞（HT22）的细胞存活率从44.06%分别提高到69.51%和76.75%。本研究为红榄李及其内生真菌的开发利用研究提供了理论基础。     

仿刺参共附生放线菌Kytococcus sp.中的环二肽成分

目的对仿刺参共附生放线菌Kytococcus sp.化学成分进行研究。方法用硅胶柱色谱,Sephadex LH-20凝胶柱色谱,反向高效液相柱层析(RP-HPLC)等分离手段对放线菌(Kytococcus sp.)乙酸乙酯提取物进行分离纯化,并利用1 H NMR、13 C NMR、质谱(MS)等手段并与文献对照相结合,鉴定化合物的结构。结果分离得到9个已知的环二肽类化合物,其结构分别为:环(L-脯-L-缬)二肽(1),环(L-脯-L-丙)二肽(2),环(L-脯-L-酪)二肽(3),环(L-脯-L-异亮)二肽(4),环(L-脯-L-亮)二肽(5),环(L-脯-L-苯丙)二肽(6),环(L-缬-L-亮)二肽(7),(L-亮-L-异亮)二肽(8),环(L-亮-L-亮)二肽(9)。结论这是首次从仿刺参中分离得到放线菌Kytococ-cus sp.,9个化合物均为首次从该属放线菌中分离得到。     

海洋放线菌Micromonospora sp.(No.69)抗MRSA活性成分研究

目的对一株小单孢菌属放线菌Micromonospora sp.(No.69)的抗耐甲氧西林金黄色葡萄球菌(MRSA)活性成分进行研究。方法采用活性追踪分离的方法,通过硅胶开放柱色谱、Sephadex LH 20柱色谱、ODS开放柱色谱、反相高效液相色谱等手段对Micromonospora sp.(No.69)发酵液的甲醇提取物进行分离和纯化,利用ESI-MS、~1H-NMR、~(13)C-NMR等波谱技术对单体化合物进行结构鉴定。结果从Micromonosporasp.(No.69)发酵液的甲醇提取物中分离得到8个化合物,分别鉴定为:环(L-缬-L-脯)二肽(1)、环(L-异亮-L-脯)二肽(2)、环(L-亮-L-脯)二肽(3)、环(甘-L-脯)二肽(4)、环(苏-L-脯)二肽(5)、环(L-丙-L-脯)二肽(6)、环(L-酪-L-脯)二肽(7)、环(L-苯丙-L-脯)二肽(8)。抗MRSA活性测试结果表明化合物1和2对MRSA具有抑制作用,IC_(50)分别为3.2 mmol·L~(-1)和6.5 mmol·L~(-1)。结论以上化合物均为首次从该属菌株中分离得到,其中化合物1和2显示出抗MRSA活性。     

中国南海蜂海绵Haliclona cymae formis的化学成分研究

目的研究中国南海蜂海绵属Haliclona cymaeformis的化学成分。方法运用正相硅胶、反相硅胶(ODS)和凝胶(Sephadex LH-20)等多种柱色谱手段对化合物进行分离纯化;通过理化性质和波谱数据,结合文献对照,鉴定化合物的结构。结果从海绵H.cymaeformis的乙醇提取物中共分离鉴定了11个单体化合物:24-亚甲基胆甾醇(1),5α,8β-epidioxy-cholesta-6-en-3-ol(2),cholesta-7-en-3β,5α,6β,9α-tetraol(3),胆甾醇(4),3-吲哚甲酸(5),尿嘧啶(6),胸腺嘧啶(7),尿嘧啶核苷(8),胸腺嘧啶核苷(9),尿嘧啶脱氧核苷(10)和胸腺嘧啶脱氧核苷(11)。结论化合物1~11均为首次从该海绵中分离得到。细胞毒活性测试结果表明,化合物3对慢性髓原白血病细胞(K562)和人肝癌细胞(SMMC-7721)的增殖显示了较强的生长抑制活性。     

海洋放线菌Micromonospora sp.(M2DG17)细胞毒活性成分研究

目的对海洋放线菌Micromonospora sp.(M2DG17)中的活性次生代谢产物进行研究。方法在活性追踪分离思路的指导下,综合利用硅胶开放柱色谱、ODS中低压柱色谱、Sephadex LH-20凝胶柱色谱以及高效液相色谱等分离技术进行化合物的分离、纯化;利用化合物理化性质和波谱学分析对单体化合物进行结构鉴定,并对其进行活性评价。结果从海洋放线菌Micromonospora sp.(M2DG17)发酵物中分离得到了7个化合物,分别鉴定为3-羟甲基-β-卡巴林(3-hydroxymethyl-β-carboline,1)、3-甲基-β-卡巴林(3-methyl-β-carbo-line,2)、β-卡巴林(β-carboline,3)、环(L-脯-L-苯丙)二肽[Cyclo-(L-Pro-L-Phe),4]、环(L-脯-L-缬)二肽[Cyclo-(L-Pro-L-Val),5]、环(L-脯-L-亮)二肽[Cyclo-(L-Pro-L-Leu),6]以及环(L-脯-L-异亮)二肽[Cyclo-(L-Pro-L-Ile),7],并对单体化合物进行了HCT116细胞生长抑制活性测试。结论化合物1~4、6为首次从该菌中分离得到,化合物2显示出弱的HCT116细胞生长抑制活性(IC50为65.0μmol.L-1)。     

海洋放线菌Kocuria sp.次级代谢产物的研究

目的 研究海洋放线菌Kocuria sp.的次级代谢产物。方法 菌株摇瓶发酵,采用现代色谱学方法(硅胶柱色谱、Sephadex LH-20凝胶柱色谱、半制备HPLC),对发酵产物进行分离,利用现代波谱学技术对化合物进行结构鉴定。结果 从海洋放线菌Kocuria sp.发酵液的乙酸乙酯萃取部分分离得到16个单体化合物：环(L-苯丙氨酸-L-脯氨酸)2 (1)、环(L-色氨酸-L-脯氨酸) (2)、环(L-色氨酸-D-脯氨酸) (3)、环(L-苯丙氨酸-D-脯氨酸) (4)、环(L-羟脯氨酸-L-苯丙氨酸) (5)、环(D-羟脯氨酸-L-苯丙氨酸) (6)、环(L-羟脯氨酸-L-酪氨酸) (7)、环(L-异亮氨酸-D-脯氨酸) (8)、环(L-亮氨酸-D-脯氨酸) (9)、环(L-亮氨酸-L-脯氨酸) (10)、环(L-苯丙氨酸-L-酪氨酸) (11)、环(L-亮氨酸-D-酪氨酸) (12)、环(L-亮氨酸-L-苯丙氨酸) (13)、环(D-缬氨酸-L-苯丙氨酸) (14)、环(D-亮氨酸-甘氨酸) (15)、环(D-异亮氨酸-甘氨酸) (16)。结论 海洋放线菌Kocuria sp.可产生结构丰富多样的环肽类化合物,所有化合物均首次从Kocuria属放线菌中分离得到。     

养殖喘水苔海绵Tedania anhelans化学成分及其生物活性研究

目的研究养殖喘水苔海绵Tedania anhelans化学成分及其生物活性,为药源海绵规模化养殖和开发提供依据。方法综合利用薄层色谱、硅胶色谱、ODS C18色谱、Sephadex LH-20凝胶柱色谱、半制备高效液相色谱(HPLC)等方法对化学成分进行了分离;通过质量光谱测定(MS)、核磁共振(NMR)等方法并结合相关文献,对化合物结构进行鉴定。结果从养殖喘水苔海绵Tedania anhelans中分离得到13个化合物,分别为胆固醇(1)、3β-羟基-24-亚甲基胆甾-5-烯-7-酮(2)、胆甾-5-烯-3β,7α-二醇(3)、胆甾-5-烯-3β,7β-二醇(4)、3β-羟基胆甾-5-烯-7-酮(5)、胸苷(6)、去甲基胸苷(7)、环(脯氨酸-缬氨酸)二肽(8)、piperazirum(9)、(E)-4-(1H-indol-3-yl)but-3-en-2-one(10)、3-吲哚乙酸甲酯(11)、3-吲哚甲醛(12)、毛脉五味子醇(13)。结论 13个化合物均为首次从该种海绵中分离得到,化合物8在50μmol·L-1浓度水平对HSV-2病毒抑制率为60.7%。     

西沙马尾藻共附生真菌Aspergillus sp.SYPUF41304次级代谢产物的分离与鉴定

目的 对西沙马尾藻共附生真菌Aspergillus sp.SYPUF41304的次级代谢产物进行分离与鉴定。方法 利用硅胶柱色谱、凝胶柱色谱、中压反相柱色谱及半制备高效液相色谱等分离手段对马尾藻共附生真菌Aspergillus sp.SYPUF41304的液体发酵提取物进行分离纯化,并利用核磁共振、质谱等波谱学方法,结合文献对照,确定化合物结构。结果 最终从提取物中分离得到10个化合物,经鉴定为吲哚-3-甲醛(1H-indole-3-carboxaldehyde)(1)、吲哚-3-乙酸甲酯(methyl indole-3-acetate)(2)、环(L-脯氨酸-L-缬氨酸)(cyclo-(L-prolyl-L-valine)(3)、brevianamide F (4)、asperterreusine A (5)、talaisocoumarin A (6)、3-甲基-6-羟基-8甲氧基-3,4-二氢异香豆素(3-methyl-6-hydroxy-8-methoxy-3,4-dihydroisocoumarin)(7)、3-甲氧基-6,8-二羟基-3-甲基-3,4-二氢异香豆素(3-met...     

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.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

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.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

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.