HIV-gp160、HIV-1 Env CTL表位肽(E7)与粘膜佐剂LT(R192G)联合鼻腔免疫的效果

大肠杆菌不耐热肠毒素 ( LT)是良好的粘膜佐剂。作者以人类免疫缺陷病毒 ( HIV)gp1 6 0及 E7为抗原与 LT突变体 R1 92 G联合投递 ,测定了对相关抗原的免疫应答及佐剂的效力。　　 2 0 μg gp1 6 0加或不加 R1 92 G5 μg,间隔1周鼻腔接种雌性 BAL B/c小鼠 ,共 3次。末次免疫后 1周采集小鼠支气管肺泡灌洗液及阴道洗液 ,以 EL ISA测定抗原特异性抗体应答。心脏穿刺采血并收集脾脏进行体外抗原再刺激试验。免疫 E7的小鼠 ,间隔 1周鼻腔接种 1 0 0 μg E7(加或不加 R1 92 G5 μg) ,共 4次。于末次免疫后 9天或 82天收获脾细胞进行铬释…     

小鼠重组乙肝病毒表面抗原鼻腔黏膜免疫效果评价

目的 评价小鼠重组乙肝病毒表面抗原(HBsAg)鼻腔黏膜免疫效果,寻找非注射乙肝免疫途径.方法 Balb/c小鼠分别经鼻腔、舌下及肌注给予重组HBsAg免疫3次,以支气管肺泡灌洗液(BALF)和血清抗HBsAg抗体水平评价3种途径的免疫效果.结果 经鼻腔免疫小鼠BALF可检测到IgA抗体,提示产生黏膜免疫:与肌注免疫类似,经鼻腔免疫小鼠血清IgG滴度大于1:500,提示产生体液免疫.结论 重组HBsAg经鼻腔免疫小鼠可同时产生体液免疫和黏膜免疫,提示鼻腔黏膜免疫可能是有效的非注射乙肝免疫途径.     

大肠杆菌LTB加痕量LT用作鼻腔接种流感疫苗的佐剂

本文讨论了大肠杆菌不耐热肠毒素B亚单位(LTB)加痕量全毒素(LT)是否可作为鼻腔接种流感疫苗的有效佐剂.Balb/c小鼠鼻腔接种1.5μg流感血凝素(HA)灭活疫苗(以流感病毒明PR8株制备),分不加佐剂组、加2μg LTB组、加痕量LT组和同时加2μg LTB及痕量LT组(2～20ng),免疫后用ELISA方法检测小鼠的IgA和IgG抗体.结果发现,前三组诱导了临界水平的抗-HA IgG抗体应答,而最后一组诱导了较高水平的抗-HA IgG抗体应答.     

可诱生粘膜IgA的新型佐剂--氧化型甘露聚糖

本文报道甘露聚糖氧化结合于重组蛋白抗原——李斯特杆菌溶血素 O融合蛋白( L LO.FP)构成的 M- LL O.FP能诱导粘膜Ig A应答。与霍乱毒素 ( CT)相比 ,氧化型甘露聚糖作为佐剂能诱导更强的抗体应答。　　作者选用 6～ 8周龄雌性近交系 ( CBA×BALB/ c) F1和 C57BL/ 1 0小鼠作为实验对象。轻度麻醉后每只小鼠鼻腔吸入 50μl(含1 2 μg抗原 )甘露聚糖抗原结合物 ,以非结合抗原作为对照。用 EL ISA测定抗体滴度。　　结果显示 :( 1 )小鼠鼻腔免疫 M- LL O.FP后可产生血清 LL O特异性 Ig A,腹腔免疫小鼠则未测得此抗体。( 2 )鼻…     

用于发展合成口服疫苗的大肠杆菌不耐热肠毒素B亚单位的表位图

不耐热肠毒素B亚单位(LT-B)含有引起LT-特异性抗体应答的大部分优势免疫表位.作者用天然LT和重组LT-B免疫小鼠,检测分泌物和血清中抗LT-B抗体应答的特性,并确定抗LT-B的T细胞或B细胞的表位图,以期构建既能刺激粘膜又能产生血清抗LT-B抗体应答的合成肽.作者从产肠毒素性大肠杆菌(ETEC)H10407株及大肠杆菌K-12株JM83转化体中分别制备LT及重组LT-B,并在都柏林沙门菌中表达重组LT-B.小鼠或于0天口服含25μgLT的磷酸缓冲盐水(PBS)0.25ml,或于0、1和2天各口服含10~(10)CFU都柏林沙门菌EL23株的PBSO.25ml,或于0、7和14天各免疫含50μg合成肽的PBSO.25ml.采集粪浸液及血清,用ELISA测定抗LT-B抗体.体外刺激LT-B特异性T细胞以评估LT-B特异性T细胞的增殖和细胞因子分布.通过细胞因子特异性ELISA检测由LT     

新型A族链球菌多表位M蛋白疫苗经注射和粘膜免疫小鼠后的免疫原性

A族链球菌 ( GAS) M蛋白是 GAS感染时的重要免疫原和毒力因子 ,是主要的 GAS候选疫苗。本文研究了 M蛋白多表位构建物——杂聚物鼻腔及口服免疫小鼠后诱导的特异性粘膜和全身抗体应答 ,并与注射接种作了比较。　　将 4～ 6周龄的 B1 0 .BR雌鼠分成 3组 ,第 1组 1 0只小鼠于第 1天鼻腔给予 2 0 μl含30μg杂聚物和 1 0μg霍乱毒素 B亚单位( CTB)的盐水 (每鼻孔 1 0 μl) ,第 7和 1 4天加强 2次 ,第 2 1天收集唾液和血液。第 2组 1 5只小鼠于第 0、7、2 8、4 2、56和 72天口服 1 0 0～ 2 0 0μl含 30μg杂聚物和 2～ 1 0μg CTB的碳…     

小鼠鼻腔免疫以大肠杆菌不耐热肠毒素非毒性突变体为佐剂的肺炎球菌多糖结合菌苗能抵抗侵袭性肺炎球菌感染

宿主抵御肺炎球菌感染很大程度上依赖由荚膜多糖特异性 Ig G抗体和补体参与的调理吞噬作用。用有效佐剂进行粘膜免疫除能诱导全身免疫应答外 ,还能诱生粘膜 Ig A抗体。大肠杆菌不耐热肠毒素 (LT)是一种研究较透彻的粘膜佐剂 ,作者以非毒性 LT突变体 LT- K6 3或 LT- R72 (两者的残留毒性很小 )为佐剂 ,用 1或 3型肺炎球菌荚膜多糖结合菌苗 (PNC)鼻腔 (in)免疫小鼠 ,研究其免疫原性。同时 ,以有毒力的同型肺炎球菌攻击来评估菌苗所产生的保护作用。　　作者将 6～ 8周远交系雌性 NMRI小鼠分成数组 (每组 8～ 1 0只 ) ,分别 in免疫或…     

纯化沙门菌鞭毛蛋白黏膜免疫诱导的系统和黏膜免疫应答

瑞典乌普萨拉大学Strindelius等采用肠炎沙门菌鞭毛蛋白、结合鞭毛蛋白的淀粉微粒和鞭毛蛋白加吸收增强剂重组霍乱毒素B亚单位(rCTB)3种配方,通过口服、皮下注射和鼻腔免疫3种途径免疫小鼠,比较黏膜免疫和系统免疫应答.     

高纯度霍乱毒素突变体E112K能诱导对白喉毒素的保护性肺粘膜免疫

为了证实鼻腔接种高纯度霍乱毒素突变体 ( m CT)能否在粘膜区室 (如肺 )和系统区室诱导白喉类毒素 ( DT)特异性保护性免疫 ,作者在小鼠中评估了 m CT作为 DT鼻腔佐剂的效力。　　用含或不含去除脂多糖 ( LPS)的 m CTE1 1 2 K或天然 CT( n CT)的 DT分别于 0、7和 1 4天鼻饲 C57BL/ 6小鼠。收集小鼠血清和粘膜洗液 ,用 ELISA检测血清和粘膜 DT特异性抗体滴度 ,用 EL ISPOT检测抗原特异性抗体形成细胞 ( AFC)。取小鼠脾和颈淋巴结 ( CL N) ,检测 DT特异性 CD4 +T细胞应答。结果显示 ,DT+佐剂组能诱导肺粘膜DT特异性 Ig A…     

6B型肺炎球菌多糖-脑膜炎球菌外膜蛋白复合物结合疫苗的结合剂量对其免疫原性的影响

为弄清多价疫苗中增加若干血清型的潜在影响 ,作者在恒河婴猴中研究了不同剂量多糖 (PS)、载体蛋白及结合物对 6 B型肺炎球菌结合疫苗 (PCV)抗体应答结果的影响。　　用 0 .0 2 5～ 2 5 μg不同剂量的 6 B型肺炎球菌 PS-脑膜炎球菌外膜蛋白复合物结合疫苗 (6 B- OMPC)于 0和 2 8天免疫婴猴两剂后显示 ,各剂量组抗 6 B型 PS Ig M抗体几何平均滴度相似 ;而抗 6 B型 PS Ig G抗体滴度则与免疫剂量呈反比 ,0 .0 2 5 μg组的 Ig G抗体滴度比 2 5μg组高 40倍。与对 6 B型 PS应答不同 ,各剂量组对 OMPC载体的 Ig G抗体应答相似 ,即使…     

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.