SARS病毒基因组所编码的E蛋白的二级结构和B细胞表位预测

目的：预测SARS病毒E蛋白的B细胞表位和二级结构。方法：以SARS病毒基因组序列为基础，采用Garnier-Robson方法、Chou-Fasman方法和Karplus-Schuhz方法预测E蛋白质的二级结构；用Kyte-Doohttle方案预测蛋白质的亲水性；用Emini方案预测蛋白质的表面可能性；用Jameson-Wolf方案预测氨基酸的抗原性指数。综合评判，预测SAPS病毒E蛋白的B细胞表位。结果：在SARS病毒E蛋白N-端的第1～6、13～19、39～43、47～64区段和第73～76区段有β-折叠中心；第6～12区段和第67～69区段可能形成转角或无规则卷曲，是柔性区域。E蛋白N端第2～13区段和第61～74区段为B细胞优势表位区域。结论：用多参数预测SARS病毒E蛋白的二级结构和B细胞表位，为实验方法探索SARS冠状病毒E蛋白的B细胞表位提供理论依据。     

人偏肺病毒黏附蛋白的二级结构及B细胞表位初步预测

目的预测人偏肺病毒G蛋白的二级结构及B细胞表位。方法分别采用SOPMA方法及HMMTOP程序预测G蛋白的二级结构和跨膜区域；综合分析蛋白的柔性结构、亲水性、表面可及性与抗原性指数，预测G蛋白的抗原表位。结果G蛋白的二级结构主要为柔性区域，占62．1％；廿螺旋占22．37％；β-折叠占15．53％；N端第32～51位氨基酸残基为跨膜区。B细胞表位位于G蛋白N端55—77、80-104、111～126、130～167、178—210区段。结论应用多参数预测G蛋白的二级结构与B细胞表位，为进一步研究蛋白特征、单克隆抗体制备及表位疫苗研制奠定了基础。     

微小隐孢子虫SA35与SA40蛋白的B细胞抗原表位预测

目的预测微小隐孢子虫SA35和SA40蛋白的B细胞表位。方法以单参数（亲水性、可及性、柔韧性及抗原性）预测为基础,通过二级结构预测初步筛选,并以ABCpred方案作为最终验证,预测微小隐孢子虫SA35和SA40蛋白的B细胞表位。结果SA35蛋白N端103～115和129～146和以及SA40蛋白的N端77～89、127～136、156～174和200～209区段为预测的B细胞表位。结论所得表位为这两种蛋白以后应用于合成肽检测、制备相应的抗体、发展高特异性和敏感性的诊断系统以及研制疫苗等提供了理论基础。     

人Izumo蛋白的二级结构及B细胞抗原表位预测

目的:预测人Izumo蛋白的二级结构及B细胞抗原表位.方法:以人Izumo基因序列为基础,按Chou-Fasman和Gamier-Robson方法预测其编码蛋白的二级结构,采用Karplus-Schulz方法预测Izumo蛋白骨架区的柔韧性;按Kyte-Doolittle方法预测其亲水性、Emini方法预测蛋白质表面可能性及Jameson-Wolf方法预测抗原性指数.结果:Chou-Fasman及Gamier-Robson两种方法预测的结果均表明,Izumo蛋白含较多的α螺旋,蛋白第6～17、30～40、88～99、103～120、153～160、173～188、249～260、283～297、334～338和339～346区段可能是α螺旋中心,第21～25、198～200、245～248和320～323区段可能是β折叠中心.用Kyte-Doolittle、Emini和Jameson-wolf方法分别对Izumo蛋白B细胞抗原表位进行预测结果表明,蛋白质第36～42、62～66、94～99、118～122、129～132、151～154、161～164、173～177、205～208、212～216、256～265、271～276、283～288、314～318和336～350区段附近很可能为B细胞表位优势区域.结论:该研究结果有助于确定Izumo蛋白的B细胞优势表位及发挥免疫避孕的活性部位.     

肿瘤相关抗原CEA的B细胞表位预测

目的分析癌胚抗原（carcinoembryonic antigen,CEA）的B细胞表位,为肿瘤治疗提供理论基础。方法以CEA的完整氨基酸序列为研究基础,采用Hopp＆Woods的亲水性方案,Emini表面可及性方案和Jameson-Wolf抗原指数方案,辅以CEA的二级结构及其柔性区域分析,预测CEA的B细胞表位。结果预测的B细胞表位可能位于CEA的N端第150～160、168～172、207～211、332～338、372～377、467～472、485～490、507～516、580～584区段。结论应用多参数预测CEA的B细胞表位,可进一步用于CEA相关肿瘤治疗性表位疫苗的分子设计和研究。     

人类偏肺病毒F蛋白的B细胞表位预测

目的 预测人类偏肺病毒F蛋白的B细胞表位。方法综合分析二级结构、亲水性、表面可及性与抗原性指数，预测F蛋白的抗原表位。结果B细胞表位位于F蛋白N段15-28、88-102、161-189、195-202、206-213、245-257、290-299、302-309、318-334、410-419、515-526区段。结论应用多参数预测F蛋白的B细胞表位，为进一步研究蛋白特征、单克隆抗体制备及表位疫苗研制奠定了基础。     

沙眼衣原体主要外膜蛋白T、B细胞多表位基因的表达及其鉴定

目的 对沙眼衣原体（Ct）主要外膜蛋白（MOMP）细胞毒性T淋巴细胞（CTL）表位进行预测和选择，并进行Ct MOMP多表位基因克隆和多表位蛋白的原核表达及其抗原性分析，为研制多表位Ct疫苗提供基础资料。方法利用SYFPEITHI法和多项式方案结合预测HLA-A2限制性的0MOMP的CTL表位，选取含多个CTL表位的基因片段作为目的基因，兼顾目的基因上下游存在的TH和B细胞表位的基因，共同组成含CTL、TH和B细胞表位的多表位基因。多表位基因经密码子优化后进行全序列合成，克隆入原核表达载体pET-32a（＋），经IPTG诱导在大肠杆菌BL21（DE3）表达并纯化，经SDS-PAGE和Western blot分析鉴定表达产物。结果 经预测筛选得到了多个MOMPHLA-A2限制性的CTL表位，成功克隆了含CTL、TH、B表位的MOMP多表位基因，并在大肠杆菌中获得了高效表达。表达产物的相对分子质量（Mr）约24×10^3，与预期肘，相符，并用Western blot方法初步证实多表位蛋白有抗原特异性。结论 成功设计了Ct MOMP的T、B细胞多表位基因，并证实在原核表达系统中获得正确表达的多表位蛋白具有良好的抗原性。     

过敏蛋白TBb的免疫活性鉴定及其表位预测

目的 对重组的苦荞过敏蛋白TBb进行免疫学活性鉴定,并预测其B细胞表位.方法 根据已获得的苦荞过敏蛋白N端结构域TBb的基因序列,构建原核表达载体pET-32m-TBb,然后转入E.coli BL21(DE3)中表达,表达产物用Ni2+-NTA琼脂糖柱亲和纯化,并用ELISA分析其免疫学活性,综合分析TBb的二级结构、亲水性、可及性、可塑性、抗原性指数,并预测其B细胞表位的分布.结果 获得了纯度95%以上的重组过敏蛋白TBb,获得的重组蛋白能与养麦过敏病人血清中的IgE抗体特异性结合,具有较高的免疫学活性,在TBb蛋白的320个氨基酸残基中,预测到的B细胞表位位于6-17,31-45,50-57,88-94,103-134,138-146,156-163,178-185,192-220,240-260,267-299区段.结论 TBb蛋白的活性分析及B细胞表位的预测为进一步研究该蛋白的分子特征及应用奠定了基础.     

人类肝素酶B细胞表位的预测和免疫原性鉴定

目的 预测并鉴定肝素酶(heparanase)蛋白B细胞表位免疫原性.方法 以肝素酶蛋白的氨基酸序列为基础,采用DNAStar分析软件以及Bcepred在线二级结构分析工具分析其蛋白二级结构并预测B细胞表位.根据预测结果 ,采用8分支多抗原肽结构合成针对该表位的抗原肽,将后者与通用型T辅助表位人IL-1β线性短肽(VQGEESNDK,氨基酸163～171)联合免疫日本白毛黑眼兔,检测免疫血清效价,鉴定其特异性和免疫原性.结果 软件预测显示,肝素酶蛋白大亚基的第1～15位(MAP1)、第279～293位(MAP2)及175～189位(MAP3)氨基酸序列最可能为其优势B细胞表位.间接酶联免疫吸附试验、免疫印迹及免疫组化分析,证实MAP1、MAP2及MAP3均能诱导机体产生高滴度抗体,但仅MAP1、MAP2抗体具有高特异性,MAP2抗体与肝癌组织的结合力最强.结论 肝素酶大亚基的第1～15位、第279～293位氨基酸为其优势B细胞表位,其中第279～293位氨基酸的免疫原性最强,这为肝素酶多肽抗体及B细胞优势短肽疫苗研制提供了理论依据.     

鼠TRAM蛋白B细胞表位预测及其免疫原性检测

目的 预测TRAM蛋白的二级结构和B细胞表位,为抗鼠TRAM单克隆抗体制备奠定基础。方法以TRAM蛋白的氨基酸序列为基础,采用Goldkeu计算机分析软件以及网络nnpredict二级结构分析软件对TRAM蛋白二级结构及B细胞表位预测。合成针对该表位的多肽,以此多肽为免疫原免疫兔,对其免疫原性进行检测。结果用多参数预测TRAM蛋白的二级结构和B细胞表位,综合评判表明：TRAM分子的第216～229位氨基酸满足亲水性、可及性和可塑性,在二级结构上位于蛋白伸展结构或无规则卷曲结构内,最可能为其优势B细胞表位。此多肽能诱导机体产生较高的抗体滴度,多克隆抗体具有高的特异性。结论TRAM分子的第216～229位氨基酸为其优势B细胞表位,这为制作B细胞优势短肽单克隆抗体提供了理论依据。     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Characteristics of the topographic interrelation of the head of the pancreas and the duodenum and pathology of the organs of the pancreatoduodenal area

Seventy pancreatoduodenal complexes of 55 patients with chronic pancreatitis and tumours of this zone and 15 patients died from other diseases are studies histotopographically . The pieces of the pancreatic head tissue in the medial wall of the duodenum were found in 12 cases of the first group and in 4 control cases. The pancreatic tissue consisted either of all elements of this organ or cystically dilated ducts and seemed to infiltrate different layers of the duodenum wall. Three variants of the pancreatic head structure are suggested on the basis of anatomo-topographical interrelationships of the pancreatic head and duodenum. In 12 out of 14 cases chronic pancreatitis and carcinoma of organs of this zone were combined with the variants of the pancreatic head structure, in 2 cases there was a true heterotopy . Pathogenetic significance of these variants for the development of chronic pancreatitis is discussed.     

Influence of the level of the transmembrane potential on the dynamics of the extinction of the amplitude of the synaptic reactions of the snail under voltage clamping conditions

The influence of the level of the transmembrane potential on the dynamics of the extinction of the amplitudes of summary excitatory postsynaptic currents arising in identified giant parietal neurons in response to rhythmic stimulation of the intestinal nerve was investigated in a preparation of the isolated CNS of the common snail in order to identify the possibility of the participation of the postsynaptic element in synaptic plasticity. It was demonstrated that, at a greater value of the transmembrane potential, the decrease in the amplitudes of the postsynaptic currents which have been induced by rhythmic stimulation takes place more rapidly. It was also demonstrated that at a higher frequency of stimulation of the nerve, the effect of the influence of the membrane potential level on the dynamics of the synaptic reactions is more prominent. The data obtained may be regarded as an argument in favor of the possibility of the participation of the post-synapse in plasticity. This study was partially supported by the Russian Basic Research Fund (project No. 94-04-12209). Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, Moscow. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 45, No. 1, pp. 171–179, January–February, 1995.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.