Comparison of new triton X-100- and tween-ether-treated split-treated vaccines in children.

Split-product vaccines (SPVs) combine the desirable properties of no systemic reactogenicity and adequate immunogenicity when two doses are given. We compared a new Triton X-100 SPV (Connaught Laboratories, Inc.) with the commercially available Tween-ether SPV (Parke-Davis & Co.) in 76 children and young adults 2 to 25 years old; there were 39 and 37, respectively, in each vaccine group. Both vaccines contained influenza A/Brazil/78, A/Texas/77, and B/Hong Kong/72 (7 microgram of hemagglutinin for each strain); two doses were administered 1 month apart. Among persons seronegative by the hemagglutination inhibition test, the geometric mean antibody titers rose to approximately 100 after the first vaccination for influenza A/Brazil/78 and A/Texas/77. For B/Hong Kong/72, however, seronegative recipients developed lower geometric mean titers of approximately 32 after one immunization. Against the new B/Singapore/79 strain neither SPV stimulated adequate cross-reacting hemagglutination inhibition antibody (geometric mean titers of approximately 10). In conclusion, the new Triton X-100 SPV appears to be comparable to the ether-treated SPV in primed subjects. Further studies in unprimed children should be done to confirm this impression. In addition, it would be advisable to study other dosage regimens in unprimed children with these SPVs.     

Molecular size and amino acid composition of H-2d antigen solubilized in Nonidet P-40.

H-2d antigenic material solubilized by the detergent Nonidet P-40 from L-1210 mouse leukemia cells was isolated by gel filtration on Bio-Gel P-100. A single peak eluted in the void volume consisted of about 90% protein, 8% hexose and traces of sialic acids. In sedimentation velocity runs, the antigen sedimented as a single peak of 3-1 S. Molecular weight determined by sedimentation equilibrium as well as calculated from amino acid composition was found to be in the range of 53,000 daltons and approx. 45,000-51,000 when calculated from sodium dodecyl sulfate polyacrylamide gel electrophoresis. Secondary structure of H-2d glycoprotein was predicted from the amino acid composition. For NP-40-solubilized H-2d antigen, about 34% of helix, 13% beta sheet and 41% turns was found.     

Nonidet P-40致流感病毒脱包膜的AFM观察研究

目的 利用原子力显微镜(AFM)观察经非离子表面活性剂Nonidet P-40(NP-40)不同浓度系列处理的A型流感病毒表面形态变化,观察不同表面活性剂-病毒表面相互作用情况,以提供一种较为温和的病毒表面裂解条件,为利用AFM进一步研究病毒下层结构提供基础.方法 用不同浓度的非离子犁表面活性剂NP-40对完整的A型流感病毒进行处理,以轻敲模式经AFM成像,获得病毒球状体和丝状体的高度图、振幅图以及相位图,并观察和比较不同浓度非离子表面活性剂对病毒表面形态和结构的影响.结果 NP-40各浓度对病度表面破坏程度不一,病毒随NP-40浓度增高而逐渐降解,并出现部分剥离病毒表面.暴露下层衣壳,更清晰地展示包膜下层表面突起的表面形态学结果.结论 通过表面活性剂优化处理病毒颗粒,实现了利用AFM观测流感病毒包膜下形态结构的设想.     

Demonstration of non-infections hemagglutinating particles of rabies virus and isolation of the hemagglutinin by disruption of the virion with Nonidet P-40

Summary Non-infectious hemagglutinating particles of rabies virus accumulated in the fluid phase of chick embryo cell cultures at 6 days post-infection, though they were undetectable at 4 days. They were characterized as looped filaments resembling viral envelope as revealed by electron microscopy.Another form of hemagglutinin (HAnin) was obtained by solubilization of partially purified virions with Nonidet P-40 (NP-40) followed by successive high speed and CsCl density gradient centrifugations. The density of the isolated HAnin averaged 1.28 g/cm³. SDS-polyacrylamide gel electrophoresis of the HAnin demonstrated that it was mainly composed of a glycoprotein (G) with a molecular weight of 83,000. Electron microscopically, it differed from the above non-infectious hemagglutinating particles, being much smaller in size and showing a star- or rosette-like appearance with a diameter of about 25 nm, composed of a central particle surrounded by particles resembling envelope-spikes. Virusneutralizing (VN) and hemagglutination inhibiting (HI) antibodies were produced in rabbits immunized with the HAnin isolated from virions.With 7 Figures     

Effect of the nonionic detergent triton X-100 on sodium permeability of the myelinated nerve fibre of Xenopus laevis

The effect of the nonionic detergent Triton X-100 (TX-100) on the Na permeability (PNa) properties of the nodal membrane in myelinated nerve fibres of Xenopus laevis was analysed with potential clamp technique. Application of TX-100 caused a rapid initial decrease in PNa that was reversible at wash out as well as a slow irreversible block. Both effects dependend on [TX-100] and duration of exposure. The reversible reduction of PNa at the steady state was 50% at 40--60 micrometer TX-100. The slope of the Hill plot for the reaction was 1.75 indicating a deviation from a first order reaction. The equilibrium dissociation constant (KD) for n = 1.75 was 0.9 X 10(-7) (M1.75). KD calculated from the rate constants for onset and offset of the reversible reaction (KD = k2/k1) was 1.5 X 10(-7) (M1.75). The possibility that the action of TX-100 involves membrane proteins is discussed.     

Nonidet P-40裂解流感病毒及其对抗体结合影响的透射电镜观察

目的 利用TEM观察经Nonidet P-40处理的A型流感病毒表面形态变化,研究不同表面活性剂-病毒表面相互作用情况,以提供一种较为温和的病毒表面裂解条件,并考察表面活性剂存在条件下对病毒表面抗体结合作用的影响,为利用病毒内部抗体结合病毒下层结构提供免疫诊断学理论基础.方法 通过用不同浓度的非离子型表面活性剂NP-40对完整的A型流感病毒进行处理,经透射电子显微镜成像,观察两种试剂对病毒表面形态的破坏程度,并考察病毒抗体结合情况.结果 NP-40各浓度对病毒表面破坏程度不一,病毒随浓度增高降解程度增大,但在0.1%NP-40及以下浓度上对抗体结合作用影响不大.结论 通过表面活性剂优化处理病毒颗粒,对抗体的结合会产生影响,本研究为利用流感病毒内层抗体结合包膜病毒奠定了样本处理的理论基础.     

Removal of unbound sodium dodecyl sulfate (SDS) from proteins in solution by electrophoresis through triton X-100-agarose

Residual sodium dodecyl sulfate (SDS) introduces artifacts into immuno- and counter- immunoelectrophoretic analysis of proteins which have been eluted from preparative SDS-polyacrylamide gels. Unbound SDS can be removed by electrophoretic passage of eluted solutions through a barrier of Triton X-100 in agarose in which the anionic and non-ionic detergents interact to form micelles.     

Ability of calf brain synaptic membranes to bind [35S]taurine to their triton X-100 and chloroform extracts.

A protein fraction containing reducing sugars was prepared from calf brain synaptic membranes with 0.5% Triton X-100, and another fraction containing bound phosphorus with a 2 : 1 chloroform--methanol mixture. Both protein fractions bound small amounts of [35S]taurine, the first fraction about 25 pmol/g protein and the second about 220 pmol/g protein. The Triton X-100 extract represented 13.8% of the membrane proteins, but the chloroform--methanol extract only 0.9%. The binding of taurine to the Trition X-100 extract was temperature sensitive, but was only slightly inhibited by hypotaurine and beta-alanine.     

Differential effect of detergents on the alkaline denaturation of haemoglobin in maternal and fetal blood, with particular reference to Triton X-100

In this investigation, the alkaline denaturation of haemoglobin in the blood of pregnant women and in cord blood obtained from newborn infants was followed by measuring the increase in absorbance at 375 nm. As expected, in the absence of detergent, the haemoglobin of cord blood was much more resistant to alkaline denaturation than that of maternal blood. However, in the presence of Triton X-100, a non-ionic detergent, the sensitivity of fetal haemoglobin to alkali was comparable to that of adult haemoglobin. Similar results were obtained using the non-ionic detergents, Brij-35, Tween 80 and Nonidet P40, but the anionic detergent, sodium deoxycholate, was apparently without effect. These findings form the basis of a rapid and sensitive method for discriminating between maternal and fetal blood in biological specimens.     

On the presence of a triton X 100 activated histidine decarboxylase activity in rat brain

Conclusion These results indicate an association between the Triton X 100 activation and the presence of a lipid membrane-bound HDD activity. The possible regulatory significance of this membrane-bound enzyme will be discussed.     

Microglial and astroglial reaction in the olfactory bulb of mice after Triton X-100 application

Gliosis including microgliosis and astrogliosis is a response to central nervous system inflammation. The purpose of this study was to evaluate whether olfactory bulbs are influenced by intranasal exposure to the detergent Triton X-100, a non-ionic surfactant. In this experiment, we measured olfactory function in mice based on the time needed to identify hidden pellets. Our results found that more time was needed to find the buried pellets by mice exposed to Triton X-100 compared with mice without Triton X-100 exposure, up to day 7. Histopathological examination revealed inflammatory cells in the olfactory mucosa and olfactory bulbs in mice treated with Triton X-100. Western blot analysis revealed significant downregulation of olfactory marker proteins in the olfactory mucosa and bulbs of mice after intranasal exposure to Triton X-100. In the olfactory bulbs of mice exposed to Triton X-100, microgliosis and astrogliosis were evident using immunohistochemistry. Cathepsin D was also upregulated in Iba-1-positive microglia/macrophages and GFAP-positive astrocytes in the olfactory bulbs of mice exposed to Triton X-100. In mice, Triton X-100 induced olfactory sensory neuron death in the nasal cavity and gliosis in olfactory bulbs with concurrent downregulation of olfactory marker protein expression, resulting in transient olfactory dysfunction.     

甘氨酸和 Triton X-100 对 ZZ-EGFP 融合蛋白分泌表达影响的初步研究

 目的 考察不同浓度的甘氨酸和 Triton X-100 对葡萄球菌蛋白质 A ZZ 亲和肽-增强型绿色荧光蛋白（ZZ-EGFP）融合蛋白在大肠杆菌分泌表达的影响。 方法 研究设计为两因素三水平析因设计。向液体培养基中分别加入终浓度 0、1%、2% 的甘氨酸和 0、1%、2% 的 Triton X-100（共 9 种组合方式，终浓度均为 0 者为对照组），诱导大肠杆菌周质腔内 ZZ-EGFP 融合蛋白泄漏到液体培养基中，以培养上清液荧光强度为观察指标，通过 ZZ- EGFP 融合蛋白浓度-荧光强度标准曲线快速检测培养基中 ZZ-EGFP 融合蛋白表达量。 结果 培养基中分别加入 1%、2% 甘氨酸或 1%、2% Triton X-100 培养后，培养上清液荧光强度分别为 283 ± 11、711 ± 19 和 622 ± 25、733 ± 25，与对照组（74 ± 5）比较，组间差异均有统计学意义（甘氨酸：F = 10.881，P = 0.024；Triton X-10：F = 12.848，P = 0.018）；而且甘氨酸与 Triton X-100两者之间有交互效应（F = 5.441，P = 0.005），其中培养基中同时含有终浓度 2% 甘氨酸及 1% Triton X-100 时培养上清液荧光强度最高（1854 ± 45），ZZ-EGFP 融合蛋白分泌表达量达到 10.4 mg/L，与对照组（0.94 mg/L）相比提高了 11 倍。 结论 甘氨酸和 Triton X-100 能提高 ZZ-EGFP 融合蛋白在液体培养基中的分泌表达量。     

Histochemical detection of glycoproteins in the gastric epithelia of Columba livia

A histochemical investigation was carried out to detect glycoproteins in the lining and glandular epithelia of 4 regions of the stomach of Columba livia: proventriculus, intermediate region between proventriculus and gizzard, gizzard, intermediate region between gizzard and duodenum. It was noticed the presence of neutral and sulphoglycoproteins, with a clear predominance of the latter in the lining epithelia and in the simple tubular glands of the mucosa of these regions, except for the gizzard. Absence of glycoproteins was noticed in the oxynticopeptic cells of the branched tubular glands of the proventriculus. Neutral glycoproteins and traces of sulphoglycoproteins were present in the lining epithelia of the folds of its central cavity and of its exit duct. Lining the above mentioned structures, the correspondent of the gizzard, inclusively there was a pellicle of sialoglycoproteins.     

Histochemical detection of glycoproteins in the intestinal epithelium of Columba livia

A histochemical investigation was carried out to detect glycoproteins in the epithelium of the small and large intestine, ceca and cloaca (coprodeum) of Columba livia. The results showed that the goblet cells of both crypts and villi of the various segments studied presented neutral and acid glycoproteins. Acid glycoproteins include sulphoglycoproteins and sialoglycoproteins with predominance of the sulphated components, particularly in the large intestine, ceca and coprodeum. In the striated border neutral glycoproteins were the predominant components in an inner wide band and the acid in a superficial pellicle.