DF—88消毒洗涤剂杀菌抗病毒效果的初步观察

本文报道一种广谱、高效且有一定洗涤作用的杀菌抗病毒溶液——DF-88消毒洗涤剂，对其杀灭常见细菌和灭活乙肝病毒的效果及其稳定性、毒性作用等进行了初步观察。结果表明，当有效碘含量为2.54mg时，能完全杀死常见细菌；有效碘含量为23mg时，可有效地灭活乙肝病毒。此外，其LD_(50)为487mg/kg，皮肤刺激试验无任何充血、红斑等改变。     

Detergent-induced epidermal barrier dysfunction and its prevention

Various detergents are used as skin cleansing products. In some cases, skin cleanser removes not only dirt but also valuable skin lipids. Therefore, detergents may disrupt epidermal barrier function despite that using of detergents are required for good skin hygiene. Lipid supplements can reverse detergent-induced dysfunction of the skin barrier. Elevated transepidermal water loss (TEWL) and riboflavin penetration in 5% SLS-treated rat and human skin were reversed by supplementation of monoglyceride (MG), squalene (SQ), cholesterol ester (CE) and pseudo-ceramide (Cer2). MG and Cer2 achieved the best results. MG appears to inhibit elution of intercellular ceramides, since more ceramides remained when the detergent was supplemented with MG. Topical application of Cer2 is not effective for recovery from artificially induced barrier disruption, but supplemented Cer2 into skin cleanser has a beneficial effect for prevention of detergent-induced barrier disruption. In conclusion, the prevention of barrier disruption is most important matter for maintaining skin health and barrier function. Therefore, we think that Cer2-supplemented skin cleanser is useful for conservation of skin barrier function.     

Acute toxicity of widely used pharmaceuticals in aquatic species: Gambusia holbrooki, Artemia parthenogenetica and Tetraselmis chuii

Pharmaceuticals are continuously dispersed into the environment as a result of human and veterinary use, posing relevant environmental concerns. This study evaluated the acute toxicity of three therapeutic agents (diazepam, clofibrate, and clofibric acid) and a detergent (sodium dodecyl sulfate; SDS) in three aquatic species, namely the euryhaline fish Gambusia holbrooki, the hypersaline crustacean Artemia parthenogenetica, and the marine algae Tetraselmis chuii. The ranking of 50% lethal concentrations (LC50) for the two animal species and 50% inhibitory concentration (IC50) for the algal species was, in decreasing order, clofibric acid > SDS > diazepam > clofibrate for G. holbrooki, clofibric acid > clofibrate > SDS > diazepam for A. parthenogenetica, and clofibric acid > clofibrate > SDS > diazepam for T. chuii. These differences show that the intrinsic nature of test organisms must be considered when evaluating the toxicity of these agents to aquatic ecosystems.     

Impact of effluents from a detergent producing plant on some water bodies in Ilorin,Nigeria

The impact of detergent-bearing effluents from a detergent-producing plant on some water quality criteria and bacterial flora of a river and its tributaries was investigated. The effluents were found to cause oxygen depletion and at some points the water bodies were completely septic. Sixteen bacterial species which belong to five families were isolated viz: Bacilliaceae, Enterobacteriaceae, Micrococcaceae, Streptococcaceae, and Pseudomonads. The effluents also appeared to altered the diversity of bacterial species in varying degrees depending on their concentration of surfactant. Species of Salmonella, Shigella, Staphylococcus, Serratia, Proteus, Streptococcus were not recovered at points below the points of effluent discharge. Results indicated an overtaxing of the water bodies by the volume of the effluents discharged.     

丙型肝炎病毒NS3蛋白酶结构域在昆虫细胞中的表达及鉴定

目的　利用杆状病毒 昆虫细胞表达系统制备丙型肝炎病毒 (HCV)丝氨酸蛋白酶结构域 (proteasedomain)的重组蛋白。方法　构建含有HCV丝氨酸蛋白酶结构域基因的杆状病毒转移载体 pFBCNS3N ,转化大肠杆菌DH10Bac进行转座 ,提取重组Bacmid用Lipofectamine法转染昆虫细胞Sf9包装成重组杆状病毒。用重组杆状病毒感染昆虫细胞进行蛋白表达 ,用SDS PAGE电泳和免疫印迹实验分析表达情况 ;利用去垢剂十二烷基肌酸钠溶解重组蛋白后以Ni NTAagrose柱亲和纯化重组蛋白 ;以重组蛋白NS5ab(包含NS5A羟基端部分和NS5B氨基端部分的重组蛋白 )为底物检测丝氨酸蛋白酶的酶切活性 ;以间接ELISA法分析重组蛋白的抗原性。结果　HCV丝氨酸蛋白酶结构域基因在昆虫细胞中获得高水平表达 ;重组蛋白能够部分溶解于含有去垢剂十二烷基肌酸钠的缓冲液 ;从 5× 10 7细胞中总共获得 0 .2mg重组蛋白 ,纯度大于 90 %;重组丝氨酸蛋白酶能够将NS5ab切割成两个片段 ;血清学实验证实该蛋白抗原性很好。结论　昆虫细胞表达的HCV丝氨酸蛋白酶结构域为膜结合型蛋白 ,具有良好的酶学活性以及抗原性。     

Effects of UVA irradiation, aryl azides, and reactive oxygen species on the orthogonal inactivation of the human immunodeficiency virus (HIV-1)

Previously we reported that hydrophobic aryl azides partition into hydrophobic regions of the viral membrane of enveloped viruses and inactivate the virus upon UVA irradiation for 2 min. Prolonged irradiation (15 min) resulted in viral protein aggregation as visualized via Western blot analysis, due to reactive oxygen species (ROS) formation, with preservation of the surface antigenic epitopes. Herein, we demonstrate that these aggregates show detergent resistance and that this property may be useful towards the creation of a novel orthogonal virus inactivation strategy for use in preparing experimental vaccines. When ROS-modified HIV virus preparations were treated with 1% Triton X-100, there was an increase in the percent of viral proteins (gp41, p24) in the viral pellet after ultracentrifugation through sucrose. Transmission electron microscopy (TEM) of these detergent-resistant pellets shows some recognizable virus fragments, and immunoprecipitation studies of the gp41 aggregates suggest the aggregation is covalent in nature, involving short-range interactions.     

RfaB,a galactosyltransferase,contributes to the resistance to detergent and the virulence of <Emphasis Type="Italic">Salmonella enterica</Emphasis> serovar Enteritidis

In this study, a deletion mutant of rfaB (ΔrfaB) was observed to be susceptible to sodium dodecyl sulfate and less tolerant to bile salts. In addition, pre-incubation in 10% bile salts increased bacterial tolerance to 30% bile salts. We also showed that the ΔrfaB mutant invaded HeLa cells less than the wild type and resulted in a lower ratio of intracellular bacteria. Competitive infection of mice showed that the ΔrfaB mutant was defective in the colonization of host organs and was cleared more quickly in fecal shedding. Transforming of a plasmid containing a wild-type allele of rfaB (pRB3-rfaB) partially rescued the defect of the ΔrfaB mutant. The results suggest that RfaB, which is responsible to add the glycosyl residue to the core lipopolysaccharide, contributes to the tolerance to detergent and the virulence of Salmonella enterica serovar Enteritidis.     

Effects of Taurine and Taurine Analogues on the Phosphorylation of a 44 kDa Protein Present in a Mitochondrial Subfraction of the Rat Heart: Partial Characterization of the 44 kDa Phosphoprotein

Recent studies suggest that taurine (2-aminoethanesulfonic acid) is involved in the regulation of protein phosphorylation in excitable tissues such as the retina, brain and heart. In order to determine the structural requirements for the effect of taurine on the phosphorylation of a 44 kDa protein(s), a series of taurine analogues were tested in an in vitro assay using a subcellular mitochondrial fraction of rat heart. Inhibitors of the phosphorylation of the 44 kDa protein include taurine and close structural analogues of taurine such as aminoethylhydrogen sulfate and α-sulfo-β-alanine. Secondary amines with the taurine structure partially locked into a saturated 5-membered ring such as (±) piperidine-3-sulfonic acid and 1,2,3,4-tetrahydroquinoline-8-sulfonic acid also possess inhibitory activity. Sulfone analogues of taurine such as 2-aminoethylmethylsulfone, a non-restricted taurine analogue with maximal conformational flexibility about its amino and sulfone moieties, and (±) 3-aminotetrahydrothiopyran-1.1-dioxide, an analogue containing the sulfone moiety in a six-membered ring structure, were found to be more potent inhibitors of phosphorylation than taurine despite the fact that the sulfone moiety is neither an isosteric nor isoelectronic substitution for the sulfonic acid moiety. The results of this study indicate that the inhibition of the phosphorylation of the 44 kDa protein in a rat heart mitochondrial fraction is relatively specific for the taurine structure. Two analogues of taurine with unsaturated rings containing a primary sulfonic acid and a secondary amine, pyridine-3-sulfonic acid and quinoline-8-sulfonic acid, were observed to be stimulators of the phosphorylation of the 44 kDa protein. In addition, 2-aminobenzenesulfonic acid also stimulated phosphorylation. Phase separation experiments with Triton X-114 suggest that the 44 kDa phosphoprotein is a soluble protein and not an integral membrane protein of the mitochondria. Phosphate incorporation into specific amino acids was determined by two-dimensional electrophoresis on celluloses plates and was found exclusively in the serine residues.     

Subtyping of human cellular prion proteins and their differential solubility

A human form of a prion disorder is the Creutzfeldt-Jakob disease. A hallmark of the disease is the accumulation of misfolded prion proteins (PrP^Sc), which exist as heterogeneous subtypes. PrP^Sc is formed by protein conversion from the host-encoded cellular prion (PrP^C), which is expressed and modified to various isoforms. Little is known about variation in PrP^C; however, it is assumed that PrP^C types play important roles in the formation of PrP^Sc. In this study, we separated distinct human PrP^C subtypes on the basis of differential protein solubilities in detergent solutions. Single and sequential application of the detergents Triton X-100, octyl-glucopyranoside and CHAPS facilitated high solubility of glycosylated PrP^C isoforms, whereas high proportions of nonglycosylated PrP^C remained non-soluble. Most proteins became highly soluble with laurylsarcosine and sodium dodecyl sulphate. Our findings demonstrate that the solubility characteristics of heterogeneous PrP^C overlap in human brains and convey distinct solubility subtypes. Differentiation by solubility experiments can therefore provide valuable information on prion protein composition, facilitate the separation of subtypes, and offer new prospects for conversion specificity of distinct isoforms.     

From assembly to virus particle budding: pertinence of the detergent resistant membranes

Detergent resistant membranes (DRMs) are the site of assembly for a variety of viruses. Here, we make use of Sendai virus mutant proteins that are not packaged into virus particles to determine the involvement of this assembly for the virus particle production. We found that, in the context of an infection, (1) all the Sendai virus proteins associated in part with DRMs, (2) mutant HN and M proteins not packaged into virus particles were similarly part of this association, (3) after M protein suppression resulting in a significant reduction of virus production, the floatation profile of the other viral proteins was not altered and finally (4) cellular cholesterol depletion did not decrease the virus particle production, although it somehow reduced their virus infectivity. These results led us to conclude that the assembly complex found in DRM fractions does not constitute a direct precursor of virus particle budding.